PMID- 2964071 TI - The donor in bone marrow transplantation. PMID- 2964072 TI - [Simultaneous detection of aflatoxin B1 and zearalenone in a mixed feed for swine]. PMID- 2964073 TI - [Cardiac natriuretic hormone (ANF)]. PMID- 2964074 TI - [Recurrent coma, papular mucinosis and benign dysglobulinemia]. AB - A woman presented with a history of three regressive comas of undetectable etiology between the age of 52 and 57 years. An IgG lambda benign monoclonal dysglobulinemia was combined with a papular mucinosis (myxedematous lichen or the generalized form of Arndt-Gotton's scleromyxedema). In the 6 analogous cases documented in the literature the onset of coma occurred generally several weeks after an aggravation of the cutaneous lesions. The coma was preceded by an influenza-like syndrome followed by asthenia, malaise with vertigo and frequently epileptic seizures. During recovery, hallucinations and transient hepatic disorders were noted. Pruritus with pronounced hypereosinophilia preceded desquamation and regression of dermatologic lesions. These comas can lead to a fatal outcome (2 of 7 cases) or regress in 2 to 20 days usually without sequelae. The disease is probably of immunologic origin. The paraprotein or a serum factor could exert a direct toxic effect on brain. As in neurologic manifestations of malignant dysglobulinemia, explained initially by a "toxic encephalosis, clinical, angiography, biologic and immunologic data exist in favor of blood hyperviscosity. This hyperviscosity could result from polymer formation through intermediates immunoglobulins and other protein chains, or again from alteration of deformability of red cells by binding of paraprotein. Hyperviscosity syndromes are frequent in system diseases that are often associated with papular mucinosis. Whatever the exact mechanism of these "comas due to papular mucinosis", a logical choice is their treatment by immunosuppressants and plasmapheresis: in the case reported, the use of plasmapheresis as soon as premonitory signs had appeared probably prevented a fourth coma. PMID- 2964075 TI - [Regional cerebral blood flow and its reactivity to acetazolamide in transient ischemic attacks. Study of 20 cases by gamma emission tomography]. AB - We have studied regional cerebral blood flow (= test) and reactivity of acetazolamide injection (= retest) in 20 patients who had presented transient ischemic attacks (TIA). The cerebral blood flow was measured by the atraumatic Xenon 133 inhalation method and single photon emission tomography using a specially dedicated tomograph (TOMOMATIC 64), allowing 3 tomographic levels (OM + 1, + 5, + 9 cm). Measurements were realized in a quiet, dark room. Acetazolamide (500 mg or 1000 mg) was injected intravenously immediately after the test, and the retest was performed 15 minutes after, in the same conditions. Results obtained from paired ROIS were analysed in terms of asymmetric index for the test, of relative reactivity for retest in regard of normal values established in a group of healthy volunteers. All patients had neurological examination, vascular explorations and CT scan. Six of the patients showed a bad reactivity (abolished or strongly decreased) which was evoked a significant hemodynamically stenosis, reflecting the loss of collateral capacity. In the other group results were diversified, and we classified these in 4 groups: "normal" (7), "luxury perfusion" (3), "limited infarction" (1), and "incomplete infarction" (3). When these groups were plotted versus duration of onset and delay between the last attack and the moment of the measurement, "incomplete infarction" clearly appears as a step between brief TIA (less of 1 hour) and "limited infarction". So rCBF and reactivity to acetazolamide are of major interest for the physiopathological classification of TIAs, but also for treatment and prognosis. PMID- 2964076 TI - Fibronectin-cell surface interactions. AB - Fibronectins are a group of closely related proteins that are found in body fluids and tissue extracellular matrices. Fibronectins play important roles in the maintenance of hemostasis and in the organization of developing tissues. The interaction of cells with fibronectin probably involves specific cell-surface molecules. Two distinct receptors for fibronectin have been postulated. One receptor, a 140-kilodalton complex, has been identified in a variety of cell types and is believed to mediate the attachment of cells to fibronectin-coated substrata. The existence of a second receptor has been proposed but it has not been identified. This receptor may function in the assembly of soluble fibronectin into the extracellular matrix. In this paper some of the recent developments in the identification and characterization of fibronectin-binding molecules on the surfaces of eukaryotic cells are outlined. PMID- 2964077 TI - The staphylococcal fibronectin receptor: evidence for its importance in invasive infections. AB - Fibronectin binds specifically and irreversibly to Staphylococcus aureus. The expression of fibronectin-binding sites correlates with the invasiveness of the isolate. It has been suggested that the fibronectin receptor of S. aureus is either a protein or a carbohydrate that localizes at the surface of the bacterium, permitting interactions of the bacteria with the fibronectin of the host. Sites at which S. aureus typically initiates infection are rich in fibronectin, e.g., blood clots and subendothelium; the cells of the luminal surface of intact endothelium are among the few cell types that are relatively devoid of fibronectin. Removal of fibronectin receptors from S. aureus by mechanical means or by growing the bacteria in an antibiotic-containing medium decreases their ability to adhere. Thus, the fibronectin receptor appears to play a role in the initiation of S. aureus infections. These findings offer hope for the development of a vaccine that will prevent invasive disease-if the S. aureus fibronectin receptor(s) can be purified, proves to be immunogenic, and inhibits bacterial adherence to host tissues. PMID- 2964078 TI - Role of fibronectin in infective endocarditis. AB - It is now generally accepted that adherence of microorganisms to various components of cardiac valve surfaces is an important early event in the pathogenesis of infective endocarditis. Several lines of evidence suggest that fibronectin may have a role in this adherence process. Fibronectin is an important component of the extracellular matrix of endothelium and fibroblasts and is exposed when these tissues are injured. Fibronectin binds to platelets and fibrin and thereby contributes to the thrombogenicity of surfaces. Because the structure of fibronectin consists of multiple functional domains, fibronectin can bind simultaneously and specifically to microorganisms and various tissue elements such as collagen and cells and may facilitate the uptake of microorganisms by endothelial cells and fibroblasts. Subinhibitory concentrations of certain antibiotics interfere with the expression of bacterial fibronectin receptors and inhibit the binding of Staphylococcus aureus to collagen matrices; this interaction may have implications for antibiotic prophylaxis of infective endocarditis. PMID- 2964079 TI - Immunoregulatory lymphokines in rheumatoid joints. II. Production by and responsiveness to interleukin-2 of eluted synovial tissue cells. AB - Since the role of interleukin-2 (IL-2) in rheumatoid synovial joints has been debated, we examined IL-2 production by, and IL-2 responsiveness of, cells eluted from synovial tissue (ST) of patients with rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). IL-2 was not detected in unstimulated cell-culture supernatants from any of the four RA patients tested, but it was present in small amounts in supernatants of unstimulated cultures derived from three of seven JRA patients studied. After PHA stimulation, IL-2 was detected in corresponding supernatants from all RA and JRA patients and from normal mononuclear cells (MNC). There was no significant difference in IL-2 activity between supernatants of normal MNC and supernatants from either RA or JRA patients. The eluted cells showed a proliferative response to recombinant IL-2. Rheumatoid ST cells are thus able to produce and respond to IL-2. Since non-T cells present in the eluates might interfere with IL-2 metabolism, one cannot yet say whether T cells of rheumatoid ST themselves produce and respond to IL-2 in a normal or abnormal way. PMID- 2964080 TI - Effects of ingestion of carbohydrate, fat, protein, and water on the mesenteric blood flow in man. AB - Transcutaneous Doppler ultrasound was used to measure superior mesenteric artery blood flow in 12 healthy volunteers in the fasting state and serially for 1 h after the ingestion of isocaloric and isovolaemic carbohydrate, fat, and protein liquid meals. The superior mesenteric artery blood flow increased significantly within 5 min of the end of each meal. The maximal responses were not significantly different but were reached at different times: carbohydrate, 64% at 15 min; fat, 60% at 30 min; and protein, 57% at 45 min. The response to the fat meal was significantly slower than the response to the carbohydrate, and the response to protein was slowest of all. In a further group of 20 fasted normal subjects no significant change in superior mesenteric artery blood flow occurred after drinking 400 ml of distilled water at room temperature. In seven of these subjects, drinking 400 ml of distilled water at 4 degrees C also did not affect mesenteric blood flow. These results indicate that the chemical nature of the meal and not the volume per se is a significant factor determining postprandial mesenteric hyperaemia. PMID- 2964081 TI - Effects of methylprednisolone on complement activation and leukocyte counts during cardiopulmonary bypass. AB - Generation of the complement activation products C3dg and terminal complement complex (TCC) and numerical changes in peripheral granulocytes (PMN) and lymphocytes were assessed in patients undergoing aortocoronary bypass surgery with extracorporeal circulation (ECC). Fluid from bronchial lavage performed preoperatively and 4 hours postoperatively was analyzed for granulocyte elastase activity and PMN content. Ten of the 20 patients received methylprednisolone (30 mg/kg b.w.) immediately before ECC. No difference was found between them and the control group regarding C3dg and TCC, and both groups showed similar postoperative decrease of peripheral blood lymphocytes. The postoperative PMN count in peripheral blood was significantly higher in the methylprednisolone group than in the controls from 12 hours onwards. In bronchial lavage fluid the postoperative PMN count was unaltered in the methylprednisolone group, but significantly increased in the controls. No granulocyte elastase activity was found before or after surgery in either group. The results indicated that methylprednisolone does not affect complement activation during cardiopulmonary bypass, but increases the granulocytes in peripheral blood postoperatively. PMID- 2964082 TI - Atrial natriuretic factor-SV40 T antigen transgenes produce tumors and cardiac arrhythmias in mice. AB - Transgenic mice that carry fusions between the transcriptional regulatory sequences of atrial natriuretic factor (a hormone intimately involved in the regulation of blood pressure) and those encoding SV40 T antigen (an oncoprotein) were generated. Although both atria express the fusion gene, the pathological response to T antigen is asymmetrical. The right atrium undergoes a several hundredfold increase in mass while the left atrium remains relatively normal in size. Hyperplasia is accompanied by a progressive increase in both the frequency and severity of abnormalities in the atrial conduction system, which ultimately result in death. PMID- 2964083 TI - A single receptor binds both insulin-like growth factor II and mannose-6 phosphate. AB - Amino acid sequences deduced from rat complementary DNA clones encoding the insulin-like growth factor II (IGF-II) receptor closely resemble those of the bovine cation-independent mannose-6-phosphate receptor (Man-6-P receptorCI), suggesting they are identical structures. It is also shown that IGF-II receptors are adsorbed by immobilized pentamannosyl-6-phosphate and are specifically eluted with Man-6-P. Furthermore, Man-6-P specifically increases by about two times the apparent affinity of the purified rat placental receptor for 125I-labeled IGF-II. These results indicate that the type II IGF receptor contains cooperative, high affinity binding sites for both IGF-II and Man-6-P-containing proteins. PMID- 2964084 TI - The Fos complex and Fos-related antigens recognize sequence elements that contain AP-1 binding sites. AB - The Fos protein complex and several Fos-related antigens bind directly or indirectly to a common sequence element that is similar to the consensus binding site for HeLa cell activator protein 1 (AP-1). This element is present in a negative regulatory sequence in the differentiation-sensitive adipocyte gene, aP2; in a transcriptional enhancer for the Gibbon ape leukemia virus; and in a region of the human immunodeficiency virus (HIV) long terminal repeat partially characterized as a negative regulatory element. The protein level and binding activity of Fos and Fos-related antigens increase rapidly after calcium ionophore treatment of a CD4+ human lymphoblast cell line, H9. These data suggest that several proteins may associate with the AP-1 binding site. Moreover, temporally regulated control of the level of each protein could represent a mechanism for modulation of these putative mediators of gene expression. PMID- 2964085 TI - Structure of the receptor for insulin-like growth factor II: the puzzle amplified. AB - The insulin-like growth factor II (IGF-II) is a polypeptide hormone with structural homologies to insulin and insulin-like growth factor I (IGF-I). In contrast to these other hormones, the in vivo function of IGF-II is not known. Although IGF-II can stimulate a broad range of biological responses in isolated cells, these responses have usually been found to be mediated by the insulin and IGF-I receptors. Recently, the receptor for IGF-II was found to also be the receptor for mannose-6-phosphate. Since this latter receptor has been implicated in targeting of lysosomal enzymes, the question is now raised of whether the same protein can also mediate metabolic responses to IGF-II. PMID- 2964086 TI - Gene encoding the beta subunit of S100 protein is on chromosome 21: implications for Down syndrome. AB - S100 protein is a calcium-binding protein found predominantly in the vertebrate nervous system. Genomic and complementary DNA probes were used in conjunction with a panel of rodent-human somatic cell hybrids to assign the gene for the beta subunit of S100 protein to the distal half of the long arm of human chromosome 21. This gene was identified as a candidate sequence which, when expressed in the trisomic state, may underlie the neurologic disturbances in Down syndrome. PMID- 2964087 TI - [In the fraternal family of the peoples of the USSR]. PMID- 2964088 TI - [Realization of Leninist ideas of the development of public health and health resorts in the Crimea]. PMID- 2964089 TI - [Public health in western Kazakhstan during the creation of Soviet power]. PMID- 2964090 TI - [Organizers of medical science and education in Azerbaijan]. PMID- 2964091 TI - [Health status and organization of medical care for industrial workers]. PMID- 2964092 TI - Gonorrheal keratoconjunctivitis in African adults. AB - We describe six cases of keratoconjunctivitis due to Neisseria gonorrhoeae in adults. All strains tested were penicillinase-producing N. gonorrhoeae (PPNG), and we conclude that the rising incidence of corneal involvement, a classic complication in the pre-antibiotic era, is due to ineffective treatment with penicillin. Indeed, the progression of the ocular lesions was stopped and microbiologic cure was obtained in all our patients after treatment with either spectinomycin or cefotaxime. These results demonstrate that early and effective treatment of adult gonococcal conjunctivitis is mandatory to avoid potentially blinding complications such as corneal ulceration and perforation of the eye. PMID- 2964094 TI - Medicare utilization by disabled-worker beneficiaries: a longitudinal analysis. AB - Medicare eligibility for Social Security disabled-worker beneficiaries begins after 2 years of cash benefit receipt. Extension of the current coverage is often proposed as a way to encourage beneficiaries to return to work. Little is known, however, about the long-run Medicare costs for the disabled and how costs vary by demographic and health characteristics. This article describes Medicare utilization and reimbursement amounts for 1974-81 for a cohort of disabled-worker beneficiaries under age 62 and first entitled to cash benefits in 1972. The data come from a first-time linkage of Disability Insurance program data with data on Medicare utilization. The tables provide a detailed look at several factors that are associated with variation in Medicare costs among beneficiaries and over time. PMID- 2964093 TI - [Comparative evaluation of the therapeutic efficiency relative to survival in hormone therapy plus cryotherapy versus hormone therapy alone in stage D2 prostatic carcinoma]. PMID- 2964095 TI - Chromosome aberrations in mixed cultures of in vitro irradiated and unirradiated human lymphocytes. AB - Human whole blood samples were exposed to different doses (1, 2, 4, 6 or 10 Gy) of gamma-radiation and mixed with different volumes of non-irradiated blood before culturing to simulate partial body irradiations. Chromosome aberrations were analysed and the frequency of dicentrics was found to be lower than expected, particularly when irradiated blood was mixed with large volumes of non irradiated blood and after exposure to high radiation doses. For the mixtures of irradiated and unirradiated blood the deviation from the Poisson distribution depends on the respective proportions and on the doses. The results can be correlated to in vivo aberration frequencies in case of therapeutical treatments, but the yields of aberrations are generally underestimated in vitro. PMID- 2964096 TI - [1st tool for division in the entire hospital]. PMID- 2964098 TI - Analysis of the antigen specific helper T cell function and HLA-DR of Israeli patients with rheumatoid arthritis (RA). AB - Forty-three patients with rheumatoid arthritis (RA) were studied for their ability to respond to the synthetic polypeptide antigen (T, G)-A-L as measured by the production of a T cell helper factor by their antigen activated T cells. Sixteen patients (37%) responded to (T, G)-A-L by the production of an antigen specific helper T cell factor, a percentage not significantly different from healthy donors. The production of antigen specific T cell helper factors was affected, although not significantly, by immune modulating drugs and by the presence of rheumatoid factor in sera of patients. The high incidence of HLA-DR 4 reported for RA patients was not observed in this group of RA patients. PMID- 2964097 TI - Emergency use of nebulised bronchodilator drugs in British hospitals. AB - A telephone survey was conducted to determine the emergency use of nebulised bronchodilator drugs by the registrar or senior house officer on duty for medical admissions at 67 British hospitals. All used a nebulised beta agonist (usually 5 mg salbutamol) as first line treatment for severe acute asthma or reversible obstructive lung disease. Twenty three doctors used ipratropium bromide occasionally and 38 used it frequently, usually mixing ipratropium and a beta agonist in the nebuliser chamber. Only five doctors routinely specified whether the nebuliser should be driven by air or by oxygen. In the case of a hypercapnic patient with chronic bronchitis, 11 respondents would not specify which gas should be used and a further 14 would use oxygen, a potentially dangerous practice. In the case of a hypoxic asthmatic patient, 22 doctors would not prescribe oxygen as the driving gas. The driving gas flow rate was almost invariably determined by nursing staff. Intravenous aminophylline was used by all 67 respondents (52 of them frequent users) but only 24 used intravenous beta agonists (five of them frequent users). It is concluded that nebulised bronchodilator drugs are the most commonly used treatment for acute asthma and reversible obstructive lung disease in hospital, but further instruction in their use is required for the staff who use them most frequently. PMID- 2964099 TI - [Relation of dental caries to infectious pathology and to a complex of sociobiological factors as well as its possible prognosis in preschool children (epidemiological research)]. PMID- 2964100 TI - [Importance of the questionnaire method for improving a lecture course on internal diseases in a dentistry department]. PMID- 2964101 TI - A reason for failure to obtain transcranial Doppler flow signals. Hyperostosis of the skull. PMID- 2964102 TI - "Plebs amat empirica": Nicholas of Poland and his critique of the mediaeval medical establishment. PMID- 2964103 TI - Evliya Chelebi's description of medicine in seventeenth-century Egypt. PMID- 2964104 TI - [The Corpus Hippiatricorum Graecorum--a disputed heritage]. PMID- 2964105 TI - [New findings on the "7-chambered uterus"]. PMID- 2964106 TI - Numisianus and Galen. PMID- 2964107 TI - Non-lymphoid thymic components tolerize T cell precursors to all the minor histocompatibility antigens of the thymus-donor strain. AB - Two months after adult-thymectomy, male B10.D2 mice were irradiated (6.5 Gy), injected with B10.D2 bone marrow cells and grafted with untreated neonate (DBA/2 X B10.D2)F1 (H-2d/H-2d) thymus [D2.F1]. Control (D2.D2) mice were implanted with B10.D2 thymus. T cells from [D2.F1] mice revealed to be fully immunocompetent while being tolerant to the minor histocompatibility antigens (MiHA) differing between DBA/2 and B10.D2 strains by in vitro and in vivo assays. Their transplantation into F1 irradiated (8.5 Gy) recipients did not induce any clinical sign of graft-versus-host reaction (GVHR) whereas this reaction is lethal when the transplantation is realized with normal B10.D2 or [D2.D2] hemopoietic cells. (D2.F1) transplanted cells were not stimulated to divide in non-lymphoid organs of the F1 hosts while this stimulation is a characteristic feature of the early period of the GVHR. Tolerance was due neither to detectable chimerism of the thymus-grafted host nor to active suppression. It is concluded that the thymus plays a sufficient and essential role for the proper acquisition of T cell immunocompetence and tolerance to all the minor histocompatibility antigens of the thymus-donor strain. Since we have previously reported that MiHA are numerous and highly organ-specific, the expression or the presence of all these MiHA in the non-lymphoid components of the thymus is questioned. PMID- 2964108 TI - Successful intestinal transplantation in pigs treated with cyclosporine. AB - To date, it has not been possible to reliably prevent intestinal allograft rejection in large animals. This study was undertaken to determine if continuous i.v. cyclosporine (CsA) followed by p.o. CsA would prevent rejection in outbred piglets with orthotopic, in-continuity intestinal allografts. Untreated recipients (n = 7) died of rejection (2), interstitial pnuemontitis (3), or technical complications (2) at 5.3 +/- 1 days. Intestinal recipients treated with i.v. CsA 8 mg/kg/day and i.v. steroids (n = 3) died of rejection (mean survival 11.3 +/- 3.2 days). CsA 20 mg/kg/day i.v. plus i.v. steroids for 21 days, followed by p.o. CsA 25 mg/kg/day (n = 6) prevented rejection; however, most of the recipients developed fatal infections (mean survival 28 +/- 8 days). Intravenous CsA 15 mg/kg/day for 7-10 days (n = 16), followed by p.o. CsA 30 mg/kg/day in tapering doses reliably prevented graft rejection, permitting long term survival (mean survival 121 +/- 32 days). Rejection did not occur in 7 animals when CsA was discontinued at 97 +/- 11 days. Seven animals surviving more than 100 days maintained normal nutritional indices and gained weight at the same rate as control animals. This study provides a rationale for further experimentation to determine the feasibility of intestinal transplantation in man. PMID- 2964109 TI - Drug-induced in vitro tolerance to allogeneic antigens. II. Further analysis of in vitro-tolerized spleen cells in a fully allogeneic murine combination. AB - C3H/HeSlc (C3H, H-2k) spleen cells were made tolerant in vitro to C57BL/6CrSlc (B6, H-2b) at the cell-mediated cytotoxicity (CMC) level by in vitro stimulation for 48 hr with mitomycin C (MMC)-treated B6 spleen cells, and treatment with 5 micrograms/ml of 5-fluorouracil for a further 9 hr. These cells were given intraperitoneally to neonate (C3HxB6) F1 mice to examine whether these tolerized spleen cells would cause lethal graft-versus-host disease (GVHD). Despite the lack of CMC, the tolerized C3H spleen cells caused lethal GVHD in most of the neonate F1 mice. Evaluating from various immune parameters, it was evident that T cell populations responsible for IL-2 production, cytostasis, and delayed footpad reaction (DFR) were retained intact after in vitro tolerance induction, probably because of their less-proliferative characteristics in response to fully allogeneic antigen stimulation, and were considered to be responsible for lethal GVHD. Contribution of natural killer (NK) cells to lethal GVHD was not ruled out. PMID- 2964110 TI - Time-response studies of the cellular immune response to cell membrane antigens. AB - It was the objective of these experiments to study the time-related changes in the responsiveness of the cellular elements of the immune system following contact with single non-H-2 or multiple H-2 histocompatibility antigens. The reactivity of spleen cells from mice that received injections of spleen cells bearing H-1c, H-3c, H-13a, or H-2b cell membrane alloantigens was characterized at intervals following antigen contact. Spleen cells taken from mice not receiving injections showed no in vitro proliferative or cytolytic responsiveness to cells bearing individual non-H-2 antigens; after in vivo antigen contact with single non-H-2 antigens there was an interval of specific cellular unresponsiveness followed by alternating periods of responsiveness and unresponsiveness. The duration of the unresponsiveness immediately following injection correlated with the strength of the injected antigen--specifically, the stronger the antigen, the shorter the period of unresponsiveness. The data indicate fluctuation in the level of helper T lymphocyte activity, as well as cytotoxic T lymphocyte activity. In contrast, in vitro responsiveness elicited by H-2b antigens with and without prior in vivo antigen contact was of a similar magnitude, and both persisted at a relatively constant level. Suppressor mechanisms were not studied. Of particular interest was the observation that in vivo contact with non-H-2 antigens resulted in suppression of spleen cell production of IL-2 in response to lectin stimulation and fluctuation in the magnitude of the primary response of cytotoxic T lymphocytes to H-2 antigens. PMID- 2964111 TI - Selective inhibition of the canine mixed lymphocyte response by HLA-DR and DP reactive monoclonal antibodies. AB - Twenty-three of 37 anti-Ia McAb reactive with human B cells, as determined by indirect immunocytofluorescence, were shown to be reactive with canine peripheral blood mononuclear cells (PBMC). Using a panel of human B cell lines that differ in their expression of HLA-DR, -DP, and -DQ molecules, it was shown that 15 of these antibodies identify HLA-DR and DP molecules (i.e., broadly reactive), while 22 identify only HLA-DR molecules. Fourteen of the 15 broadly reactive McAb were reactive with canine PBMC while only 9 of the 22 HLA-DR-specific McAb reacted with canine PBMC, suggesting that broadly reactive anti-Ia McAb are much more likely to react with canine cells than narrowly reactive McAb. Ten of the canine reactive McAb that were shown to identify typical Ia bimolecular structures on canine cells using immune precipitation analysis were tested for blocking activity in the canine mixed lymphocyte culture (MLC). All four of the broadly reactive McAb (B1F6, J-70, 9-49, and HB10a) plus two of the six narrowly reactive McAbs (H81.98.21 and H40.164.3) blocked the canine MLC when added to culture wells on day 0, suggesting that inhibition may be related to the specificity of the anti-Ia McAb employed. Since the MLC may reflect cellular interactions occurring during graft-versus-host disease, this assay may be useful for screening functionally relevant broadly reactive McAb in experimental canine bone marrow transplantation studies. These data suggest that the dog may be a useful model to study anti-Ia immunotherapy. PMID- 2964112 TI - Lymphoid resistant responses to in vitro immunosuppression in renal transplant recipients: increased incidence of refractory episodes. PMID- 2964113 TI - Cellular basis of passive enhancement of graft survival. PMID- 2964114 TI - [Prevalence of chronic ORL diseases among schoolchildren of Tbilisi]. PMID- 2964115 TI - [Prevalence of chronic ear, throat and nose diseases among children and their need for otorhinolaryngological care]. PMID- 2964116 TI - [Toxic-allergic reaction in a patient after hemilaryngectomy for cancer of the larynx]. PMID- 2964117 TI - Cryptosporidium infections in birds and mammals and attempted cross-transmission studies. AB - Infections by Cryptosporidium were detected in association with clinical disease in 11 humans (Homo sapiens), 19 calves (Bos taurus), nine common quail (Coturnix coturnix), six mallard ducks (Anas platyrhynchos), five ring-necked pheasant (Phasianus colchicus) and a single budgerigar (Melopsittacus undulatus). Infections in mammals were accompanied by transient diarrhoea and anorexia, whereas infected birds exhibited clinical signs of respiratory distress. Repeated cross-transmission studies revealed apparent strain differences or differences in the host specificity of several mammalian and avian isolates for homologous vertebrate classes only. Oocysts from humans and calves were infective to mice, pigs or lambs, but not to chickens, whereas oocysts from quail and pheasant were infective to chickens, but not to mice. PMID- 2964118 TI - Hosts of two canid genera, the red fox and the dog, as alternate vectors in the transmission of Sarcocystis tenella from sheep. AB - Microscopic sarcocysts recovered from naturally infected sheep were infective to both the domestic dog (Canis familiaris) and the red fox (Vulpes vulpes). The parasite was passaged through experimental specific-parasite-free (SPF) sheep three times: infection was transmitted twice with sporocysts from foxes and subsequently with sporocysts from dogs. The sarcocysts from sheep muscle were infective to both dogs and foxes on each occasion. A cat was not infected. The prepatent period in individual canids ranged from 7 to 15 days. Sporocyst excretion was still detectable 60 days post infection. This study establishes that canids of two genera may act as vectors for a single isolate of the same Sarcocystis species from sheep. PMID- 2964119 TI - [Treatment of hypertension with a delayed-action form of verapamil and its effect on hypertrophy and function of the left ventricle]. PMID- 2964120 TI - [Vitamin PP and C allowances and their correction in the treatment of bronchial asthma patients]. AB - Providing with vitamins PP and C was studied in 124 patients with bronchial asthma (59 males and 65 females). Deficiency of the vitamins was recorded in the majority of the patients during the disease exacerbation, especially in the winter-spring period of the year. Decreased providing with nicotinic acid was noted in 83%, with ascorbic acid in 96.8% of the patients. The treatment without vitamins did not lead to normalization of providing with vitamins. Applying vitamin loadings lasting many days, rational doses of vitamins PP and C were determined for the correction of providing patients with these vitamins in the period of bronchial asthma exacerbation. Depending on the severity of their state the patients should be given 90-110 mg of nicotinic acid and 275-300 mg of ascorbic acid/day, during the course of the treatment. Inclusion of rational doses of vitamins PP and C intensifies the therapy effectiveness and reduces the terms of the inpatient treatment. PMID- 2964121 TI - [Energy requirements of workers engaged in the underground repair of oil wells in the Azerbaijan SSR]. AB - The energy requirement of workers engaged in the underground repair of oil wells has been evaluated. The energy expenditure during the 8-hour shift with the use of manual and mechanized labor was determined on the basis of the 7-day chronometric charts of their labor activity. The power consumption of 32 descending-lifting and preparative-conclusive operations was evaluated by the indirect calorimetry. Energy expenditure in the time off was studied by the questionnaire method during a week. It was established that, as it was calculated for standard body mass (70 kg), the daily energy expenditure of an operator comprised 3190 kcal (fluctuations depended on the use of mechanization means, 3125-3398 kcal), of his assistant--3487 kcal (3434-3644 kcal), machine driver of the tractor-elevator--3013 kcal (2965-3100 kcal). On the basis of the data obtained the work of the operator and machine driver has been referred to the III, and that of the operator's assistant to the IV occupational group with respect to the intensity of their labor. PMID- 2964122 TI - [Energy requirements of miners engaged in open-pit coal mining]. AB - It has been found that according to the value of daily energy consumption the coal miners engaged in open-cast mining should be referred to group III-IV of work intensity. The determining component of daily energy consumption is energy consumption during the working period the value of which depends on the character of working activity and duration of the working shift. The method of strict evaluation of the calorific value of the food taken and the control of body mass is acceptable for the assessment of daily energy consumption. PMID- 2964123 TI - [Atrial natriuretic hormone (review of the literature)]. PMID- 2964124 TI - [Allergic reactions to fibrinolysin]. PMID- 2964125 TI - [The correlation between the E. coli gene nmpC and the bacteriophage gene LC]. PMID- 2964126 TI - [Clinical evaluation of Berodual in patients with bronchospastic syndromes]. PMID- 2964127 TI - [Therapy of prostatic cancer with cyproterone acetate]. AB - 45 patients with recently detected prostatic cancer were treated with cyproterone acetate (CPA) at a dosage of 100 mg/die. 25 patients underwent primary orchiectomy and consecutive treatment with 200 mg CPA/die. In 44.4% of the 45 patients who underwent monotherapy with 100 mg CPA/die progression of cancer was observed 10.8 months after beginning of treatment on average 55.6% of these patients showed a remission or stabilisation. 7 patients died, 4 certainly due to prostatic cancer. In the 25 patients who underwent orchiectomy and treatment with 200 mg CPA/die, 92% showed a remission. One patient of this group died, but not as a result of cancer. Based on the present data it may be concluded that treatment of inoperable prostatic cancer with 100 mg CPA as monotherapy is not indicated in patients with poorly differentiated tumours. Combination therapy--in addition to other possibilities of contrasexual therapy in prostatic cancer- seems to be of high efficacy. PMID- 2964128 TI - [Nonsurgical reconstructive treatment of peripheral arterial occlusive disease]. PMID- 2964129 TI - [Therapy of peripheral occlusive disorders in diabetes mellitus]. PMID- 2964130 TI - [Current contact allergens]. AB - We present the patch test results of 2,623 patients treated at the Department of Dermatology, University of Heidelberg, from July, 1984 to June, 1986. One test at least was positive in 680 patients (25.9%). 257 allergens produced a total of 1,450 positive reactions. (average: 2.1 per patient). 9.7% of the allergens caused 64.4% of the positive reactions. The 5 most frequent allergens were nickel sulfate, balsam of Peru, formaldehyde, neomycin sulfate, and cobalt sulfate. Mixed fragrances and Kathon CG were among the 20 most frequent allergens though only tested during half of the study period. Glyceril monothioglycolate was the leading allergen with hairdressers. Other allergens clinically relevant were rubber gloves, bufexamac, bronopol, and propolis. PMID- 2964131 TI - [Current problems in expert assessment]. AB - Problems concerning occupational dermatology usually arise before the actual expert opinion is given. Preventive measures should be taken into consideration very carefully in order to reduce the risk of an occupational dermatosis to a minimum. Further problems are accurate methods of patch testing, the formal wording of the expert opinion, and the assessment of the degree of disablement. PMID- 2964132 TI - [Anaphylactoid reaction to trypsin following local administration of Dontisolon ointment]. AB - We report on a female patient with known epicutaneous allergies, who developed type I allergy after oral local application of Dontisolon ointment. Epicutaneous tests with constituents of the ointment proved a positive reaction to trypsin-S calcium glutaminate. We refer to similar cases of allergy to trypsin and chymotrypsin reported in the literature. PMID- 2964133 TI - [Significance of local factors for the development of contact allergic reactions in patients with chronic venous insufficiency]. AB - There are a number of local factors which might contribute to the induction and persistence of contact allergies in patients with chronic venous insufficiency of the lower leg and their sequelae such as stasis dermatitis and leg ulcers. Thus, the acanthotic epidermis might have a certain "reservoir function" for potential contact allergens. Furthermore, the density of epidermal Langerhans' cells is increased in areas of stasis dermatitis, and the dermis contains a high number of cells bearing HLA-DR antigens, which might act as antigen-presenting cells. PMID- 2964134 TI - [Remarks on contact eczemas--etiopathogenetic considerations based on immunohistochemical studies]. AB - Functionally defined inflammatory cells can be selectively labeled in tissue sections by immunohistochemical methods. We investigated the cellular infiltrate in acute toxic contact dermatitis and various forms of contact eczema and other inflammatory dermatoses. In all cases, the so-called "superficial T-cell pattern" was evident, which comprised T-lymphocytes, predominantly helper/inducer cells, as well as Langerhans cells/indeterminate cells. The numerical density of these cell types, which represent the cellular prerequisites for delayed hypersensitivity reactions, turned out to be markedly increased in affected skin. Compared with normal skin, therefore, lesional skin provides a different immunological microenvironment. We conclude that a negative patch test reaction in normal skin does not exclude the possibility that the same allergen may provoke and enhance immunological reactions in eczematous skin. Eczema is regarded as an abnormal reaction of the organism towards exogenous factors; the abnormal reaction may be confined to sites with a considerable infiltrate. This might explain the pronounced chronicity of contact eczemas, even after withdrawal of the presumed cause, and the existence of chronic allergic contact eczemas without positive patch test results. The inflammatory infiltrate may be interpreted as a major predisposing factor for further immunological reactions at the same site. PMID- 2964135 TI - [The formaldehyde problem]. AB - Regarding formaldehyde (FA), we face the following problems: (1) It is a wide spread substance being found as free FA, in FA releasers and FA resins. (2) FA contact allergy occurs either as occupational or non-occupational disease. (3) In spite of its frequency there are still problems concerning patch testing of FA. We report on the results of patch testing with FA in various vehicles and concentrations performed on patients with dermatitis and those suffering from a proven FA contact allergy. Our investigations confirm the superiority of water as a vehicle. PMID- 2964136 TI - [Contact allergies to antifungal agents]. AB - Contact allergies to modern antimycotic agents are comparatively rare considering the high prescription rates. They may be caused by components of either the vehicle or the active ingredient. According to both the literature and our own investigations, the most important sensitizers of the vehicle are cetylstearyl alcohol and propylene glycol; as to the active substances, miconazole and nystatin are the most frequent sensitizers of the imidazole and non-imidazole group, respectively. Cross-reactions of imidazole derivatives mainly occur between miconazole and econazole. For patch testing with imidazoles, we recommend a 1% alcoholic solution of the active ingredient. PMID- 2964137 TI - [Galenic quality of test substances]. AB - According to experimental investigations (microscopy, analytical determination, method of agar diffusion, spreading on human skin) there is no inhomogeneity detectable among the 20 test substances of the Standard-Epicutantest-HERMAL. The following criteria are proved: equal spectrum of the grain sizes, equivalent contents of the active substance along the whole syringe, good release of the active substances from vaseline, homogeneous spreading of the active substances in the syringe, homogeneous spreading on human skin, when Finn Chambers on Scanpor (FCS) and Epiquick plasters are applied. These findings meet the requirements for relevant and reproducible test results. PMID- 2964138 TI - [Use of a new prepared patch in general practice (Epiquick)]. AB - We report on an improved technique of epicutaneous testing. The Finn chambers of the Epiquick system are already loaded with the test material. In a study on 36 patients, the new system was compared with the conventional patch test. The results of both techniques corresponded well for 9 allergens (72.7% at 72 h). Further investigations must settle whether the discrepancies found with some allergens are related to the new system. The simple handling of Epiquick can save considerable time in the patch testing procedure. PMID- 2964139 TI - [Transluminal coronary angioplasty: parameters of increased risk of recurrence]. AB - Data about the primary success and complication rates following transluminal coronary angioplasty are comparable worldwide. Reports on the frequency of recurrences, however, differ considerably. Data concerning factors enhancing the risk of restenosis are contradictory. We compared a group of 62 patients with angiographically confirmed restenosis with a group of 62 patients without restenosis. There were no significant differences with regard to age, sex, onset of symptoms and severity of angina pectoris, number of previous infarctions, incidence of multivessel disease, maximum size of the balloon catheters used and primary dilatation results. The following factors were significantly associated with the occurrence of restenosis: high-grade stenoses prior to dilatation, stenoses involving long segments of coronary arteries, eccentric stenoses, the necessity to apply high balloon pressures, long duration of balloon inflation, and also the patient being overweight, a nonsmoker and having undergone an alteration in continuous treatment. PMID- 2964140 TI - [On the editorial of U. Gleichmann and P. Lichtlen: Percutaneous transluminal coronary angioplasty: qualifications of the surgeon and patient security, logistical considerations]. PMID- 2964141 TI - PTCA--clinical aspects. AB - At Emory University Hospital, 7,000 coronary angioplasty procedures have been performed since 1980. The overall success rate remains around 90% despite extensive broadening of the indications. Complications include emergency surgery in 2.5%, acute infarction in 2.5%, and a mortality of 0.2%. Major problems are acute closure and recurrence of stenosis. Factors predisposing to acute closure include lesion length, female gender, bend in the artery at the area of stenosis, branch point location, thrombus in the artery and other lesions in the same vessel. Therapy for acute closure includes prolonged balloon inflations, the use of the bail-out catheter, and for the future, intracoronary stenting devices. Restenosis occurs in approximately 25% of the patients and is influenced by male gender, unstable angina, total occlusion, location of the lesions, and incomplete results of angioplasty. Attempts at modifying restenosis included anti-platelet therapy, anticoagulation and calcium antagonists. At the present it is felt that more potent antiplatelet agents will be required to make additional improvements in the restenosis rate. Long-term follow-up of the Emory cohort reveals a 4 1/2 year survival of over 96% with 89% free from death, myocardial infarction or the need for coronary bypass surgery. In order to identify the proper role of angioplasty in multivessel disease, a prospective randomized trial is now being instituted at Emory to evaluate the relative merits of angioplasty and coronary bypass surgery. PMID- 2964142 TI - The sliding rail system (monorail): description of a new technique for intravascular instrumentation and its application to coronary angioplasty. AB - The sliding rail technique is a new technique for intravascular instrumentation, especially coronary stenosis dilatation. The so-called monorail balloon catheter is the first device which can be used according to this technique. The monorail catheter has a single lumen shaft and only a short central tube within the distal balloon part. With the guidewire inserted into the tube, the balloon can be advanced or retracted on the guidewire as on a sliding rail. The most relevant improvements for coronary dilatation are steerability, contrast flow and rapid and easy exchangeability of balloon catheters and other intracoronary devices. These characteristics are felt to result in a more simple and time- and fluoroscopy-saving dilatation procedure. A special transfusion catheter may also improve procedural safety. The first clinical results in 69 patients with a success rate of 96%, an emergency bypass rate of two patients (one infarction) and a stenosis improvement of 58% confirm the theoretically conceived advantages. PMID- 2964143 TI - Methods for the assessment of PTCA success. AB - Numerous criteria for the assessment of the immediate and late results of percutaneous transluminal coronary angioplasty (PTCA) are currently in use. Here, the values and limitations of the transstenotic gradient (TG), visual qualitative angiogram assessment, quantitative coronary angiography and the coronary flow reserve (CFR) will be discussed. Although measurement of the TG may be clinically useful, current data suggest that it does not accurately reflect the "true" pressure-flow-resistance across coronary lesions. Furthermore, the widely applied method of visual interpretation of coronary angiograms is hampered by several serious shortcomings: large intra- and interobserver variabilities, and lack of correlation with pathologic as well as intraoperative findings. In contradistinction, CFR and minimal luminal cross-sectional area (MLCA) appear to be more reliable parameters for judging the physiologic importance of (residual) coronary obstructions. In fact, given the curvilinear relation between CFR and MLCA (r = 0.92), the available evidence suggests that at the moment quantitative coronary angiography--with measurement of the MLCA immediately after PTCA--is the method of choice in assessing the efficacy of coronary angioplasty. PMID- 2964144 TI - Platelet-vessel wall interactions in the development of restenosis after coronary angioplasty. AB - Acute occlusion and restenosis of the dilated coronary segment remain serious unsolved complications that can occur after coronary angioplasty. Acute occlusion is seen in approximately 3-5% of patients and restenosis in 13-47% of patients undergoing this procedure. While the causes of these complications are incompletely understood, platelets and their interactions with the vessel wall appear to be important. The mechanism of stenosis dilatation involves a splitting or tearing of the atherosclerotic plaque as well as desquamation of the endothelial layer of the vessel wall. Therefore, superficial as well as deep arterial injury occurs as a result of angioplasty. Acute occlusion and restenosis appear to be the biological response to this injury. Following even subtle injury to the endothelial layer, platelets adhere to the vessel wall and become activated, releasing substances such as platelet derived growth factor which stimulates intimal hyperplasia and re-growth of the atherosclerotic plaque. Deeper injury results in increased platelet deposition and mural thrombus formation. Rheologic factors including oscillatory shear forces and high local shear rates resulting from residual stenosis and intimal dissection further promote platelet activation and thrombus formation. Both the rheologic factors and the thrombogenic properties of the vessel wall and circulating blood promote restenosis and acute occlusion. Studies using animal models have demonstrated that platelet inhibitor drugs used in addition to heparin can reduce the amount of platelet deposition at the time of angioplasty, and may have a favorable effect on the occurrence of acute occlusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964145 TI - Coronary angioplasty guidewire technology. AB - The technology of coronary guide wires has advanced rapidly since Simpson and Robert performed the first angioplasty with a movable guidewire system in 1981. The technology involved in engineering guidewires encompasses three areas: (1) selection of materials, (2) design of the configuration, and (3) development of the construction process to fabricate the guidewire. Each of these areas must be considered concurrently; that is, the functional effects of manufacturing a wire must be considered along with material selection and configuration design. Another critical area is functional testing to ensure safe and effective performance of the guidewire in the clinical setting and to confirm its functional limits. Research is directed continually toward advancing existing guidewire technology to improve functional performance and develop more efficient manufacturing processes. Additionally under development are new guidewire devices incorporating microsensors and fiberoptics to provide real-time feedback. Advances in technology require miniaturization of guidewire components and application of new materials and processes to manufacture new designs. PMID- 2964146 TI - Dilatation balloons: polymer selection, balloon design and assembly. AB - The current status of coronary dilating instruments is presented from the designer's perspective. Catheter shaft design is considered by important features, common catheter materials and types of catheter construction. Among the seven companies manufacturing balloon dilating instruments, only three types of catheter and four materials are offered. Balloon design is presented by important features and by materials selected. Performance comparisons are made between the three materials used to fabricate dilating balloons: PVC, PE and PET. PMID- 2964147 TI - The use of pressure-volume curves to assess the mechanism and adequacy of stenosis dilatation by balloon angioplasty. AB - This report introduces a new technique for monitoring and understanding the mechanical events taking place during coronary balloon angioplasty. A standard inflation syringe is instrumented with transducers to record intra-balloon pressure and volume during stenosis dilatation. The theoretical and practical aspects of obtaining and interpreting PTCA "pressure-volume" curves are described. Based on mechanical principles, this method may provide the PTCA operator with information regarding the stiffness and resilience of each stenosis, the mechanism of dilatation, and a measure of adequacy of lumen enlargement even before the balloon is deflated. PMID- 2964148 TI - Long wire technique--experience with 100 procedures. AB - Derived from more than 1000 procedures the long wire technique has proven to facilitate coronary angioplasty. Maneuvering of the wire is unhindered because the wire is introduced without balloon catheter. Optimal contrast display is possible during crossing of the stenosis. Precision and safety of the procedure is therefore considerably improved. Balloon catheters can be exchanged without recrossing the stenosis including super low profile catheters of small diameter. In case of acute coronary occlusion occurring during angioplasty the long wire allows reintroduction of catheters without danger of via falsa. If necessary a 4.5 f perfusion catheter can be introduced and coronary perfusion with 60-100 ml per minute blood from the femoral artery can be performed by hand. In contrast to the monorail technique the ability to measure pressures through the balloon catheter including intracoronary gradients and coronary capillary pressure is preserved. PMID- 2964149 TI - [Ultrastructural and electron-histochemical changes in the mouth mucosa in soft leukoplakia]. PMID- 2964150 TI - [Improving the methods of health education of various groups among urban and rural populations]. PMID- 2964151 TI - [Problem of the prevention of occupational dermatoses induced by organic dyes]. PMID- 2964152 TI - [Experience with the treatment of patients with onychomycosis caused by Trichophyton rubrum]. PMID- 2964153 TI - [Complex treatment of patients with psoriasis at the Naftalan health resort]. PMID- 2964154 TI - [Transluminal extraction of intracardiac embolized catheter fragments in children]. AB - The frequent use of central venous catheters for administration of fluids and especially ventriculovenous shunts in children sometimes lead to embolisations of catheter fragments. Because of the potential complications, extraction of the fragments is mandatory. In 5 children aged 1.8 to 7.8 years fragments were removed from the right heart and the pulmonary artery by use of a loop snare, a helical basket or a myocardial biopsy catheter. In four cases the fragment was the distal part of a ventriculoatrial shunt, in one case the central part of a silastic catheter had accidentally advanced into the pulmonary artery. In two cases adhesions of the fragment to the wall of the pulmonary artery impeded the extraction. PMID- 2964155 TI - A retrospective study of 91 cases with gastroschisis or omphalocele 1956-1985. AB - The experiences of treatment of 91 infants with abdominal wall defects during thirty years are presented. The occurrence of gastroschisis increased during the last ten years. The results of treatment have improved markedly during the period of study but there is still a high mortality rate in children with omphalocele and multiple concomitant malformations and in children with gastroschisis and coexisting intestinal atresia. In cases with large omphalocele defects, if primary closure is not possible, an implant is recommended instead of silastic silo, skin flap closure or mercurochrome application. The clinical course in thirty-six patients during the last ten years is reported. PMID- 2964156 TI - Siderophore production by enterotoxigenic Salmonella. AB - Eleven Salmonella strains of diverse origin studied previously for enterotoxigenicity, cell surface hydrophobicity (HIC) and haemagglutination (HA) were studied for the production of a siderophore precursor 2,3-dihydroxybenzoic acid (DHB) by colorimetry. The Salmonella strains were similarly grown aerobically and anaerobically in a deferrated (DF) medium containing 0.05 micrograms/ml iron at 22 degrees C and 37 degrees C. Salmonella strains positive for both heat-labile (LT) and heat-stable (ST) enterotoxins, i.e. ST+ LT+ strains produced 25-70 mg/l, DHB, bound to hydrophobic phenyl sepharose gel and exhibited mannose-resistant haemagglutination (MRHA) of bovine, chicken and human group A erythrocytes. Conversely, heat-labile enterotoxin-positive, only i.e. ST- LT+ strains produced less than 25 mg/l DHB and were negative for the three aforementioned parameters. This dichotomy was shown by strains grown aerobically at 22 degrees C. Strains grown at 37 degrees C gave disparate results while Salmonella anaerobically grown at both growth temperatures did not produce DHB. These studies demonstrate production of a siderophore during iron stress. PMID- 2964157 TI - [Spectinomycin sensitivity of Neisseria gonorrhoeae]. AB - Using the agar dilution technique minimal inhibitory concentrations (MIC) with respect to spectinomycin were determined. 92 Neisseria gonorrhoeae isolates collected in a STD outpatient clinic and 160 Neisseria gonorrhoeae isolates collected from prostitutes were investigated. One isolate imported from Asia had a MIC of greater than 1024 micrograms/ml and was resistant. All other isolates, three penicillinase producing strains included, had MICs of 4-32 micrograms/ml and were sensitive to spectinomycin. PMID- 2964158 TI - Protective immunity against Vibrio cholerae infection in the rabbit. AB - The DIC model (Duodenal Inoculation with ligation of the Cecum in rabbits) was employed to study experimentally induced cholera and the related protective immunity. Duodenal inoculation (DI) without ligation of the cecum with live V. cholerae organisms did not cause any disease symptom but induced protection against subsequent challenges with homologous and heterologous organisms for up to 24 months. After 30 months this protective immunity began to decrease. A similar protective immunity could be induced by administration of the A- B+ derivative CVD101 of V. cholerae strain 395. This type of experiment can only be done successfully with conventional, healthy rabbits held under low stress conditions. A so-called specific pathogen-free rabbit breed was found to be entirely unsuitable. Duodenal inoculation with heat- or merthiolate-inactivated V. cholerae for a prolonged period of time by means of an intestinal osmotic minipump did not induce protection. Injection of heat-inactivated V. cholerae material into the Peyer's patches sometimes led to protection, suggesting that a thermostable antigen, possibly lipopolysaccharide, is one of the major protective antigens. Duodenal administration of a combination of inactivated V. cholerae serotypes Ogawa and Inaba cells and 1 mg B subunit of the V. cholerae enterotoxin by up to three inoculations protected only 3 out of 12 rabbits against challenge. The results obtained on the rabbit model are discussed in relation to the efficacy of this vaccine in human volunteers and in a recent field test. PMID- 2964159 TI - [Mercuric chloride-induced enteropathy in the rat: a model of gastrointestinal disease with immunologic pathogenesis]. AB - Protein-loss into the gastrointestinal tract is a feature of many different diseases, a number of which are immunologically mediated, e.g. systemic lupus erythematosus or Crohn's disease. The pathogenic mechanism of protein-losing enteropathy in diseases without obvious enterocyte injury are unknown. Brown Norway-rats (BN-rats) were gavaged with low doses of HgCl2 for 39 weeks. Within 2 weeks, the intestines showed strong linear staining for IgG and IgA along vascular and intestinal basement membranes. Granular deposits containing IgG and C3 were present along intestinal basement membrane only in the late stages of the experiment; only in these animals increased intestinal protein loss as measured by fecal Cr-51 excretion was found. These findings suggest that immune complex deposition along the intestinal basement membrane can lead to protein-losing enteropathy. This disease may be used as a model for the study of immunologically mediated intestinal disease. PMID- 2964160 TI - [Human atrial natriuretic factor (hANF) in liver cirrhosis]. AB - Mean plasma levels of hANF at admission were significantly higher in liver cirrhosis (LC) patients with ascites (93 +/- 11 ng, n = 20; p less than 0.05) than in LC-patients without ascites (32 +/- 14 ng/l, n = 11) or healthy controls (31 +/- 15 ng/l; range: 5-80 ng/l; n = 106). Diuretic treatment of patients with LC and ascites normalised hANF plasma concentrations (44 +/- 14 ng/l; p less than 0.05). Increase of plasma hANF in LC-patients with ascites after acute volume expansion was lower (relative rise: 159%) than in those without ascites (relative rise: 223%). Volume redistribution into the vascular compartment, induced by peritoneovenous shunt implantation in LC-patients with diuretic treatment refractory ascites, resulted in a sharp increase of plasma hANF levels (344 +/- 87 ng/l = 420% of preoperative concentration). Normal and slightly increased levels of plasma hANF and the appropriate regulation to volume changes suggest an intact control of the hormone in LC-patients. Neither a lack nor an inappropriate secretion of hANF as a cause for sodium retention as postulated by the overflow theory of ascites accumulation could be found in liver cirrhosis. PMID- 2964161 TI - Local x-ray irradiation of the rat head (30 Gy). II. Behaviour and effect of meclophenoxate after 24, 72 and 120 hours. PMID- 2964162 TI - Local x-ray irradiation of the rat head (30 Gy). III. Behaviour and effect of pyritinol after 24, 72 and 120 hours. PMID- 2964163 TI - Assessment of the Ohio 5400 volume monitor Ohmeda SE302 spirometer and the flow meters of the Servo 900C ventilator. AB - The performance of the Ohio 5400 electronic anemometer and the Ohmeda SE302 heated platinum filament respirometer were assessed in a series of bench tests in which they were incorporated into the patient circuit of a Siemens-Elema Servo 900C ventilator. The reference instrument was a Fleisch pneumotachograph. The Ohio 5400 was accurate to within 10% when measuring air flows between 23 and 44 l/min, provided the cycling frequency did not exceed 30 b.p.m. and the flow wave form was not accelerating. The Ohmeda SE302 was accurate to within 10% when measuring air flows between 8 and 48 l/min, regardless of cycling frequency or shape of respiratory wave form. Both instruments displayed marked inaccuracies (of the order of 20-30%) when measuring 100% oxygen or 66% nitrous oxide/oxygen mixtures. The performance of the flow transducer of the Servo 900C was also assessed. This was significantly superior, and its output was virtually identical to the uncorrected output of the pneumotachograph. PMID- 2964164 TI - Effects of an organosilicon compound on the tubular apparatus of rat kidney. A histological and enzyme histochemical report. AB - Male adult Wistar rats were treated for 8 weeks with an organosilicone [2,2 dimethyl-4-(chloromethyl)-1,3-dioxa-2-silacyclopentane] employing two different dosages (25 and 50 mg/kg body weight i.p. daily) and the changes in the tubular apparatus of the kidney were investigated employing histological and enzyme histochemical methods. The effects, more pronounced at the higher dosage, were the following: The brush borders of the proximal tubules were stuck together and disintegrated; only few epithelial cells remained intact showing a decreased activity of nonspecific esterases and the increase of beta-hydroxybutyric acid dehydrogenase. The nuclei of most of the epithelial cells in the distal tubules were dislocated towards the enlarged lumen and the cytoplasma showed a decrease of nonspecific esterases and an increase of beta-hydroxybutyric acid dehydrogenase. The collagen fibers in the walls of the collecting tubules were dislocated and disintegrated with a discontinuous border of Mg2+-ATPase; the nuclei of the epithelial cells were pyknotic, the cytoplasma showed an increase of beta-hydroxybutyric acid dehydrogenase. The induced changes were partially reversible after a recovery period of 8 weeks. PMID- 2964165 TI - Clinical studies with a new, low-dose oral contraceptive combination, containing 0.150 mg desogestrel and 0.020 mg ethinylestradiol. PMID- 2964166 TI - Dopamine D2-receptor affinities of resolved C1-dimethylated 2-aminotetralins. PMID- 2964167 TI - A C2-methylated derivative of the 5-hydroxy-tryptamine-receptor agonist 8-hydroxy 2-(dipropylamino)tetralin (8-OH DPAT). PMID- 2964168 TI - Skeletal muscle biopsy studies of cardiac patients. AB - Eleven patients diagnosed and treated for congestive cardiomyopathy (COCM) of unknown aetiology, and another 10 patients, with congestive alcoholic heart muscle disease (ACOCM) were studied. Muscle biopsy samples were obtained from the vastus lateralis (VL) and the gastrocnemius (G) muscles. In part of the sample muscle the fibre pattern was classified by means of ATPase activity staining, a technique based on the pH lability of the fibres concerned. Fibre typing and area measurements were carried out by light microscope. The other part of the sample was used as muscle homogenate of which the Ca2+-activated ATPase activity as well as citrate synthetase (CS) and aldolase activities were measured. No significant difference was found in these enzyme activities between the two groups of patients. The proportion of the slow twitch (ST) fibres in the VL, mainly in the patients with ACOCM, was lower as compared to data for healthy subjects. A similar tendency was revealed for G. In both muscles tested, the area of ST fibres was smaller in the ACOCM group. The fast twitch (FT) fibre area proved to be slightly different in the two groups of subjects tested. Occurrence of degenerative signs in the histological tests was higher in the ACOCM than in the COCM group. It was concluded that differences in the skeletal muscles of patients with ACOCM and COCM may primarily account for the alcoholism. The disease of the heart muscle has little effect on the function of skeletal muscle. Even so, a low amount or lack of physical activity may have an unfavourable influence on the skeletal muscles of patients with heart muscle disease. PMID- 2964169 TI - Effects of anticurare agents produced in Bulgaria on the smooth muscles of the gastro-intestinal tract. AB - The effect of nivalin-P (nivalin:pymadin/4-aminopiridine hydrochloride ratio- 1:1) on the smooth muscles of the gastro-intestinal tract was studied. The experiments were carried out on guinea-pig ileum, rabbit jejunum using the method of Magnus, and on isolated strips of guinea-pig stomach and taenia coli by the modified method of Golenhofen. The agents studied were added to the incubation medium in the following concentrations: acetylcholine--1 X 10(-7) g/ml to 2 X 10( 6) g/ml, pymadin--up to 2 X 10(-6) g/ml, nivalin--up to 2 X 10(-6) g/ml, nivalin P--up to 2 X 10(-6) g/ml, pancuronium, diadonii and galamine--from 2 X 10(-5) to 4 X 10(-4) g/ml, isoptin--from 1 X 10(-5) to 2 X 10(-5) g/ml and sodium nitroprusside--2 X 10(-5) to 4 X 10(-5) g/ml. Nivalin was found to increase mainly the tone of the smooth muscles, while pymadin increases predominantly the amplitude of the contractions. The combined agent nivalin-P stimulates both the phasical and the tonic activity. The role played by calcium ions in the mechanisms of the stimulating effect of nivalin-P on the smooth muscles is discussed. PMID- 2964170 TI - [ATPase activation in Bacillus subtilis strain 168 with phospholipids]. PMID- 2964171 TI - Ca-inhibitory myosins: their structure and function. AB - Actin-myosin-ATPase interaction of Physarum is under inhibitory Ca-control. Ca binding to myosin inhibits the interaction, while Ca-release from myosin removes inhibition. However, this myosin-linked control system alone is not sufficient to account for in vivo contraction. A Ca-dependent inhibitory factor, whose active principle appears to be identical with or similar to Ca-Lc of myosin, augments myosin-linked control by interacting with actin. This novel regulatory system may not be specific for the actomyosin system in Physarum plasmodia, because some actomyosin-related processes, i.e., secretion and motility, are subject to inhibitory Ca-control. Accumulation of examples for the inhibitory Ca-control in higher and lower eukaryotes will clarify its phylogenic significance. PMID- 2964172 TI - Providing motor control for the handicapped: a fusion of modern neuroscience, bioengineering, and rehabilitation. PMID- 2964173 TI - Physiology of normal and abnormal blinking. AB - Blink rate is determined by many factors, including local eye irritation, the state of the corneal tear film, factors related to general visual function, the amount of general facial movement, cognitive variables, and the level of arousal. These factors appear to be mediated by several neuroanatomic structures (Table 2). The timing and the nature of the interrelationship between neuroanatomic structures during blinking remains to be determined. Dopamine is the neurotransmitter that is most strongly linked to blinking, exerting its effect on blinking primarily through the D2 receptor. The reduced rate in Parkinson's disease seems to implicate the nigrostriatal system. Perhaps efferents of the nigrostriatal system, such as those to the superior colliculus, are primarily involved, as suggested by the reduced blinking in PSP. Changes in blinking produced in the sylvian aqueduct syndrome further suggest involvement of the periaqueductal structures. At best, however, these conclusions are tentative, as the biochemical neuroanatomy will probably prove more complicated than suggested by the initial studies using the dopaminergic paradigm. Nevertheless, insofar as blink rate represents a noninvasive probe of CNS dopamine activity, the failure to associate dyskinesias (except levodopa-induced dyskinesia) with increased blinking, indicates that the pathophysiology of these conditions may not involve hyperactivity of CNS dopamine systems. Fittingly, the current clinical potential of blink rate seems maximal in parkinsonism, both to follow the severity of the illness and to monitor side effects of dopamine agonist treatment. PMID- 2964174 TI - Dysphagia in movement disorders. PMID- 2964175 TI - [Allergologic-immunochemical study of tree and bush pollen. II--Study of the sensitization spectrum of patients with seasonal rhinitis in the spring]. AB - 80 patients sera were investigated by means of radioallergosorbent test (RAST) and enzyme linked immunosorbent assay (ELISA). Specific IgE antibodies against ten various tree pollens were determined. There is a significant correlation between RAST and ELISA results. It was found that birch, beech, alder, hazel and oak pollens are most important as causes of springtime hay fever. 75% of patients had increased specific IgE-titres against these pollens whereas maple, poplar, elm, sallow and ash allergens more often gave negative or only weak positive test results. PMID- 2964176 TI - [Allergologic-immunochemical study of tree and bush pollen. III--Cross reactions of human IgE antibodies with various tree pollen allergens]. AB - Partial identity between the major allergens of birch, beech, alder, hazel and oak pollen extract could be identified by means of RAST-, ELISA- and CRIE inhibition as well as further types of crossed immunoelectrophoresis. It seems that spring time pollinosis in our region of Central Europe is caused principally by the major allergen of birch pollen. Cross reactivity between grass and tree pollen could not be found. Patients with symptoms in spring and summer time are double sensitized. PMID- 2964177 TI - Phagocytosis of cadmium microcrystals by glass-adherent peripheral blood leucocytes of newborns with various perinatal risks. AB - Using cadmium microcrystals saturated with human serum albumin as particles for the phagocytic assay, the phagocytic activity of peripheral blood glass-adherent leucocytes of 32 preterm infants, 15 small-for-date newborns, 14 term newborns with perinatal asphyxia and 27 term newborns of diabetic mothers was compared with healthy term newborns and normal adults. A statistically significant increase in both the percent of phagocytosing cells and in the mean number of engulfed particles per cell was found in all groups of "stressed" infants as compared with cells of adults. No difference in the proportion of actively phagocytosing cells was found between term healthy and stressed neonates; as regards the mean number of ingested particles per cell, a slight increase (however, statistically not significant) was found in all groups of infants with perinatal risks compared with normal infants suggesting a temporary stimulation of these cells. PMID- 2964178 TI - In vitro immunization for antibody production against tetanus toxin and toxoid. 3. Recombinant gamma interferon and indomethacin support specific antibody production induced by antigen-pulsed immobilized monocytes. AB - Specific antibody production against Tetanus Toxin by in vitro cultured human peripheral blood lymphocytes may be enhanced, when monocytes were immobilized onto FCS-coated culture plates and then pulsed with Tetanus Toxoid. Using this model several immunomodulators were tested for their influence on in vitro immunization efficiency. A combination of recombinant gamma interferon and indomethacin added during antigen presentation to lymphocytes, was able to potentiate specific antibody response. PMID- 2964179 TI - [Optimal production of murine monoclonal antibodies in ascites of syngeneic mice by a single whole body irradiation]. AB - Hybridoma cells injected intraperitoneally into mice induce formation of ascites tumors producing ascites fluid with high levels of monoclonal antibodies. Several parameters affect the growth of the immunoglobulin-producing tumors in vivo. In the present study the average ascites tumor formation rate of 10 different hybridomas could be increased from 32% (n = 338 mice) to 77% (n = 112 mice) by only one whole body irradiation of paraffin-pretreated-Balb/c mice. Production of monoclonal antibodies was better in males because significantly (p less than 0.01) increased volume of ascites fluid. From the increased tumor formation rate in irradiated mice it is suggested that in non-irradiated recipients the tumor growth rate was lowered by immunological reactions against hybridoma cells provoked by cell surface neoantigens revealed by cell fusion and/or tumor associated antigens of the myeloma parent cells as well as by altered antigen pattern caused by possible mutations in the myeloma cell line and/or Balb/c/K strain. PMID- 2964180 TI - [Interleukin 2 inhibitor activity in blood serum]. AB - Normal murine, horse and human sera inhibit the proliferation of thymocytes, T blast cells and cloned cytotoxic T lymphocytes in vitro dose dependently. This effect is specific to IL 2-dependent cell proliferations. The inhibitory activity does not decrease the binding of IL 2 to its receptor, but inhibits the IL 2 receptor expression dose dependently. An excess of purified IL 2 can overcome the inhibition of IL 2-dependent cell proliferation. Using gel filtration, we have found the inhibitory activity in the region of 80,000 MW. This inhibitory activity is precipitated by 50% saturated ammonium sulfate. The species specificity of this inhibitor in serum is very low. The IL 2-inhibitor is discussed as a part of natural immunoregulation and as a hint for activation of the immune system. PMID- 2964181 TI - Technology and science in the management of deafness. PMID- 2964183 TI - Coronary dissection: a predictor of restenosis? AB - To determine if arterial dissection resulting from transluminal coronary angioplasty (TCA) leads to a greater incidence of restenosis, 273 consecutive patients who had undergone TCA with at least 1 year of follow-up were examined. Success was graded as a greater than or equal to 20% increase in intraluminal diameter. Dissection was defined as a prominent intimal defect at the site of angioplasty. Restenosis was defined as a 50% loss of the initial angiographic gain. Of 216 patients in whom TCA was successful, 64 (30%) had dissections at the site of angioplasty and 152 (70%) had no dissection. During follow-up, 135 patients were asymptomatic with normal results of exercise tolerance tests, recurrent symptoms developed in 81 patients, and 64 patients underwent repeat angiography. The overall restenosis rate was 21%. In the dissection group, 11 (18%) had documented restenosis and 49 (82%) did not develop restenosis. In the nondissection group, 35 (24%) had documented restenosis and 113 (76%) did not develop restenosis. These data show that patients who develop dissections at the time of TCA are no more likely to develop restenosis during follow-up. PMID- 2964182 TI - Coronary angioplasty immediately after thrombolysis in 115 consecutive patients with acute myocardial infarction. AB - Between September 1981 and May 1986, coronary angioplasty immediately after intracoronary thrombolysis was attempted in 115 patients with acute myocardial infarction. The present study describes our experience with this combined procedure. Primary success was achieved in 102 patients (89%). Before discharge, 79 of these patients agreed to be restudied angiographically. The infarct-related vessel was still patent in 71 patients (patency rate of 90%). Sequential left ventricular angiograms of quality sufficient to allow automated analysis were obtained in 58 patients. Global ejection fraction improved significantly from 52 +/- 10% to 55 +/- 9% (p = 0.01) from the acute to the chronic stage. In patients with anterior infarction, the increase in global ejection fraction was primarily the result of significant improvement of the regional myocardial function of the infarct zone. No significant changes in global and regional myocardial function could be seen in patients with inferior infarction. However, when patients in whom the infarct-related vessel was reoccluded at follow-up angiography are excluded from analysis, the global and regional myocardial function did improve significantly irrespective of the location of the infarct. Median clinical follow up of 20 months (range 4 to 50) resulted in an overall mortality rate of 4%. Preservation of global and regional left ventricular function with a low mortality rate suggests that immediate coronary angioplasty after thrombolysis can be safely used to provide reperfusion in the setting of acute myocardial infarction and that this combined procedure may be the optimal mode of therapy. Further randomized studies are warranted to precisely define the role of coronary angioplasty in acute myocardial infarction. PMID- 2964184 TI - The resolution of coronary collaterals after successful percutaneous transluminal coronary angioplasty. AB - It has been shown that collaterals can develop rapidly during acute coronary occlusion, either due to thrombosis or during angioplasty (PTCA). However, the fate of well-developed collaterals immediately after a successful PTCA is unknown. Accordingly, 15 patients with Rentrop class 2 or 3 collaterals as visualized angiographically were studied immediately after successful single vessel PTCA. The left anterior descending artery contained the stenosis in nine patients and the right coronary contained the stenosis in six patients. There was total occlusion of six vessels and subtotal occlusions of nine vessels pre PTCA. Immediately after PTCA, flow through the collaterals to the stenosed artery could no longer be visualized angiographically in eight patients (group 1), but remained faintly visible in seven patients (group 2). There was no difference between these two groups with regard to pre PTCA transstenotic pressure gradient (46 +/- 12 vs 42 +/- 14 mm Hg), post PTCA pressure gradient (13 +/- 7 vs 11 +/- 10 mm Hg), or post PTCA percent luminal diameter narrowing (26 +/- 18% vs 24 +/- 13%). These findings suggest that despite similar hemodynamic and angiographic improvement, the resolution of collaterals immediately after PTCA is variable. PMID- 2964185 TI - Relationship between location of chest pain and site of coronary artery occlusion. AB - Chest pain characteristics and site of coronary artery occlusion were evaluated in 148 patients having single-vessel coronary angioplasty and in 95 patients having double-vessel angioplasty. The locations of chest pain included substernal and left precordium, right precordium and epigastric. The possible sites of pain radiation were limited to neck/jaw, left arm, right arm, and interscapular. The patient described whether or not the pain was typical of previous angina, and the presence of ST segment deviation was noted to be certain that ischemia was present. The analysis showed that the occluded artery could not be reliably identified. However, it was possible to say which artery was most likely not diseased. Patients presenting with substernal or left chest pain with radiation to the left arm had a less than 10% chance of having right coronary artery disease. A patient presenting with epigastric pain radiating to the neck or jaw had a less than 13% chance of having left anterior descending disease. It is concluded that in patients with single- and double-vessel coronary disease, there is some relationship between chest pain pattern and disease location. PMID- 2964186 TI - A practical application of Doppler echocardiography for the assessment of severity of aortic stenosis. AB - This study evaluated a strategy that makes optimal use of Doppler echocardiography for estimating the severity of valvular aortic stenosis (AS). Fifty-eight patients with no more than moderate aortic insufficiency who underwent cardiac catheterization were evaluated with two-dimensional echocardiography and Doppler velocimetry to determine the peak velocity across the stenotic valve and aortic valve area (AVA) by means of the continuity equation. All 33 peak Doppler velocities of greater than or equal to 4 m/sec had critical AS (AVA less than or equal to 0.8 cm2 at catheterization). Conversely, six of seven patients with Doppler velocities of less than or equal to 3 m/sec had noncritical AS. The patient with a falsely low peak velocity had severely depressed left ventricular function. Doppler velocity alone was inadequate in determining severity of AS for patients with velocities between 3 and 4 m/sec. The continuity equation proved accurate in estimating AVA in the 46 patients for whom catheterization and ultrasound data were sufficient to compare calculated AVA (r = 0.81), and was also accurate for those patients with peak Doppler velocities between 3 and 4 m/sec (r = 0.90). These results suggest that Doppler velocimetry alone is adequate in determining critical vs noncritical AS in many patients, while the continuity equation should be applied for patients with peak velocities between 3 and 4 m/sec as well as in patients with severely depressed cardiac function. PMID- 2964187 TI - Thrombolytic therapy for femoral artery thrombosis after pediatric cardiac catheterization. AB - Femoral artery thrombosis remains a well-known complication after cardiac catheterization. A study was undertaken to assess the efficacy of thrombolytic therapy for this complication. A total of 526 consecutive infants and children were prospectively evaluated after cardiac catheterization, and the medical charts of 42 patients who required femoral artery thrombectomy between 1975 and 1985 were reviewed. In the prospective study, patients were given a bolus injection of heparin, 150 U/kg, at the time the artery was entered. Patients with persistently absent or diminished pulse 2 hours after catheterization received a second bolus injection of 50 U/kg followed by an infusion of 20 U/kg/hr heparin for a maximum of 48 hours. If the affected leg pulse was absent or reduced and the systolic Doppler blood pressure was less than two thirds that of the unaffected leg, thrombolytic therapy was begun. In the 42 patients with surgical thrombectomy, there were no serious complications of surgery. Forty-five of the 526 patients (8.6%) had a decreased or absent pulse after catheterization. Of these 45 patients, 32 (71.1%) improved with systemic heparinization only. Thirteen patients (28.9%) had a persistently absent pedal pulse suggesting femoral artery thrombosis, despite continuous heparinization. Eleven patients were successfully treated with thrombolytic therapy and two required surgical thrombectomy. Intraarterial balloon dilatation procedures were performed in 8 of these 13 patients. Prothrombin time was prolonged (11.5 +/- 1.06 to 52.3 +/- 40.4 seconds; p less than 0.025) and fibrinogen levels were significantly reduced (2.25 +/- 0.79 to 1.52 +/- 0.52 gm/dl; p less than 0.01) during therapy. There were no serious complications, although four patients bled from the groin entry site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964188 TI - Balloon angioplasty for coarctation of the aorta: immediate and long-term results. AB - Twenty-five infants and children with native coarctation of the aorta had percutaneous balloon angioplasty over a 28-month period ending in May, 1987. The mean systolic pressure gradient across the coarctation decreased from 47.6 +/- 20.9 mm Hg to 10.3 +/- 7.3 mm Hg (p less than 0.001) following angioplasty, and the diameter of the coarcted segment increased from 3.2 +/- 1.7 mm to 7.8 +/- 3.5 mm (p less than 0.001). Clinical and echo-Doppler follow-up indicated excellent results in 16 of the 18 patients in whom 3- to 22-month follow-up was available; two infants required additional treatment (repeat angioplasty in one and surgical resection in the other). Fourteen patients who underwent repeat cardiac catheterization remain improved with regard to pressure gradient across the dilated coarctation (9.5 +/- 9.6 mm Hg, p less than 0.001) and angiographically measured sizes of the coarcted segment (10.3 +/- 3.2 mm, p less than 0.001). No aneurysm was seen in any child. We recommend balloon angioplasty as the therapeutic procedure of choice for relief of severe, previously unoperated coarctation of the aorta in neonates and young infants. Routine use of balloon angioplasty for unoperated coarctation of the aorta in children appears indicated, but should await longer follow-up results and reports of follow-up on a larger number of patients; this caution is mainly based on reports from other workers of aneurysm formation at the site of balloon dilatation. PMID- 2964189 TI - Simultaneous percutaneous aortic valvuloplasty and coronary artery angioplasty in an elderly patient. PMID- 2964190 TI - Assessment of mitral flow velocity with exercise by an index of stress-induced left ventricular ischemia in coronary artery disease. AB - Exercise-induced myocardial ischemia results in both diastolic and systolic left ventricular (LV) dysfunction. To investigate the utility of Doppler assessment of LV diastolic function with exercise, 28 consecutive patients underwent digital stress echocardiography, including measurement of mitral flow velocity by pulsed wave Doppler and simultaneous stress thallium imaging. The mean mitral flow velocity was measured as the integrated area under the LV diastolic inflow Doppler spectral display. The change in mean mitral flow velocity from baseline to immediate postexercise was compared among 3 patient groups: (1) patients with thallium redistribution or exercise-induced wall-motion abnormalities, or both, consistent with exercise-induced ischemia (n = 18), (2) patients with no evidence of stress-induced ischemia, with or without resting wall-motion abnormalities (n = 10), and (3) 10 control subjects of similar age with normal resting 12-lead electrocardiograms, normal resting and postexercise 2-dimensional echocardiograms and normal electrocardiographic treadmill stress testing. The percent increase in mean mitral flow velocity was 101% (+/- 59) for controls and 86% (+/- 53) for patients without stress-induced ischemia versus 33% (+/- 24) in patients with stress-induced ischemia (p less than 0.005). An increase in mean mitral flow velocity with exercise of greater than 50% correctly identified 9 of 10 nonischemic control patients. An increase in mean velocity of less than 50% predicted ischemia in 15 of 18 patients with evidence of stress-induced ischemia (p less than 0.005) Thus, Doppler assessment of LV diastolic function with exercise expressed as a change in the mean velocity of mitral flow is a useful indicator of stress-induced ischemia. PMID- 2964191 TI - Dynamics of early and late left ventricular filling determined by Doppler two dimensional echocardiography during percutaneous transluminal coronary angioplasty. AB - To study the influence of ischemia on both early and late left ventricular filling, Doppler 2-dimensional echocardiography was used to measure filling parameters during percutaneous transluminal coronary angioplasty (PTCA) in 26 patients. Doppler recordings were taken immediately before balloon inflation and continuously during balloon inflation and deflation until 60 seconds into the recovery period. During PTCA of the left anterior descending artery (16 patients) there was a 35% decrease in early peak filling rate from 269 +/- 88 to 169 +/- 55 ml/s (p less than 0.0005) by 60 seconds of balloon inflation. In patients undergoing PTCA of the left circumflex (4 patients) or dominant right coronary artery (6 patients), the early peak filling rate decreased 15% from 325 +/- 126 to 284 +/- 137 ml/s (p less than 0.005). The decrease in early peak filling rate became evident at approximately 15 seconds after balloon inflation and fully recovered 20 seconds after balloon deflation. Rather than an expected increase in atrial stroke volume and a decrease in early to late filling ratio during coronary occlusion, there was a 28% decrease in atrial stroke volume during left anterior descending coronary artery PTCA and a 6% decrease during right coronary and circumflex PTCA. Because of the simultaneous decrease in both early and late ventricular filling, peak early to late filling ratio was only slightly altered during PTCA. There was an 83% increase in mean pulmonary artery wedge pressure during balloon inflation from 12 +/- 5 to 20 +/- 4 mm Hg. In 11 of these patients global systolic function was measured on subsequent inflations during PTCA using 2-dimensional echocardiography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964192 TI - Early and late assessment of stenosis geometry after coronary arterial stenting. AB - Early and late modifications of stenosis geometry after stenting of coronary arteries were assessed. Morphologic changes were evaluated by quantitative coronary angiography (automated edge-detection) and theoretical pressure drop across the dilated and stented stenosis was calculated from the Poiseuille formula, with turbulent resistances assuming a coronary blood flow of 1 or 3 ml/s. Eleven patients (ages 41 to 69 years, mean 55) were studied before and after angioplasty, and immediately after stent implantation. The stented coronary artery was the left anterior descending artery in 9 patients and the left circumflex in 2. Following stent implantation, an additional increase in minimal luminal cross-sectional area of the dilated vessel was observed, suggesting that the self-expanding stainless-steel endoprosthesis used had a dilating function in addition to its stenting role. Repeat angiography in 6 patients 3 months after stent implantation showed a decrease in the minimal luminal cross-sectional area without a significant change in theoretic pressure decrease. This slight reduction in vessel caliber had no hemodynamically significant repercussions. Thus, stenting of coronary arteries following dilatation is a potentially valuable technique for preventing both abrupt closure and late reduction in lumen diameter. PMID- 2964193 TI - Effect of mitral regurgitation on diastolic filling with left ventricular hypertrophy. AB - Earlier studies have suggested that mitral regurgitation (MR) augments early left ventricular (LV) diastolic filling. To determine whether MR affects early diastolic filling in patients with abnormal diastolic filling, transmitral pulsed wave Doppler recordings were used to study 32 normal subjects, 21 patients with LV hypertrophy, 23 with LV hypertrophy and MR and 15 patients with MR. Patients with MR had increased peak early filling velocities (MR 108 +/- 27 cm/s, normal 80 +/- 16 cm/s, p less than 0.01), peak atrial filling velocities (MR 72 +/- 18 cm/s, normal 55 +/- 12 cm/s, p less than 0.05) and increased deceleration rates (MR 5.0 +/- 1.9 m/s2, normal 3.5 +/- 1.2 m/s2, p less than 0.05). Patients with LV hypertrophy had reduced peak early filling velocities (69 +/- 14 cm/s, p less than 0.05) and increased peak atrial filling velocities (83 +/- 16 cm/s, p less than 0.001). There was also an increase in the atrial filling fraction and reduction in the rapid filling fraction as compared with normal patients. Patients with LV hypertrophy and MR had increased peak early filling velocities (98 +/- 26 cm/s, p less than 0.01 vs normal, p less than 0.001 vs LV hypertrophy patients), increased atrial filling velocities (84 +/- 27 cm/s, p less than 0.001 vs normal), increased deceleration rates (4.4 +/- 2.4 m/s2, p less than 0.05 vs normal) and a normal distribution of diastolic filling. Within the LV hypertrophy and MR group, diastolic filling parameters were similar when patients were subgrouped on the basis of auscultability of MR. MR augments early diastolic filling and may tend to normalize diastolic filling patterns in LV hypertrophy patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964194 TI - Optimum heart rate of large failing hearts. PMID- 2964195 TI - Effect of right coronary artery occlusion during percutaneous transluminal coronary angioplasty on right ventricular performance. PMID- 2964196 TI - Sudden death and cardiomegaly unassociated with coronary, valvular, congenital or specific myocardial disease. PMID- 2964197 TI - The effect of different methods of bracket removal on the amount of residual adhesive. AB - The amount of residual adhesive following three different methods of bracket removal was examined. The effects of tooth position, bracket base type, and composite mix were also examined. It was found that there was a nonsignificant trend for more composite to remain on upper anterior teeth and a highly statistically significant difference between a paste/paste and paste/liquid composite mix. Bracket base type showed no significant difference. One method of bracket removal resulted in significantly less composite remaining than the other two methods. PMID- 2964198 TI - Skeletal anomalies in trisomy 21 as an example of amplified developmental instability in chromosome disorders: a histological study of the feet of 21 mid trimester fetuses with trisomy 21. AB - In a previous radiographic study on the feet of 71 adults with trisomy 21 we found, in comparison to control individuals, an increased prevalence of biphalangeal toes and metatarsophalangeal sesamoid bones. The present histological study on the feet of 21 mid-trimester fetuses with prenatally diagnosed trisomy 21 confirms results of the earlier study. At both stages of development these minor bone anomalies have about the same frequency, thus suggesting 1) that they are selectively neutral, and 2) that they reflect a basic (innate) failure of ordered morphogenesis. Our observation that the normal spatial pattern of skeletal variants is reproduced in trisomy 21 simply on a quantitatively higher level lends sound support to the hypothesis of amplified developmental instability in chromosome trisomies. PMID- 2964199 TI - Specificity versus nonspecificity in the pathogenesis of aneuploid phenotypes. PMID- 2964200 TI - Percutaneous transcatheter recanalization in the management of acute renal failure due to sudden occlusion of the renal artery to a solitary kidney. AB - Percutaneous angioplasty was attempted in 5 patients with acute renal failure due to occlusion of the artery to a solitary functioning kidney. Angioplasty was technically successful in all patients. Renal function was completely restored in 3 and renal perfusion improved in 1 patient. In the fifth patient, renal function did not return despite ultrasound and radionuclide scan evidence of renal reperfusion. Transcatheter thrombolytic therapy was attempted in 2 patients, 1 of whom also underwent angioplasty. In this patient, perfusion was restored to most of the kidney. In the other patient an infrarenal aortic occlusion was present. During thrombolytic therapy, intrarenal microembolization occurred from lysis of the aortic thrombus, leading to irreversible renal damage. PMID- 2964201 TI - Antagonists of the antidiuretic activity of vasopressin. AB - Competitive antagonists of the antidiuretic (ADH) activity of vasopressin were first described some six years ago. When studied in vitro, ADH antagonists displace vasopressin from specific renal binding sites and antagonize, in a competitive fashion, vasopressin stimulation of adenylate cyclase and transepithelial water, salt, and urea fluxes. When studied in vivo, the ADH antagonists increase renal water excretion and antagonize, in a competitive fashion, the ADH activity of vasopressin. Marked species heterogeneity is apparent with ADH antagonists in vivo, and inconsistencies between in vitro and in vivo findings within the same species are reported. Other renal responses associated with administration of ADH antagonists include changes in renal hemodynamics and renal salt and urea excretion. The effects on salt excretion appear to be limited to those species in which vasopressin stimulation of epithelial salt reabsorption has been demonstrated. In summary, the role of vasopressin as the principal factor regulating renal water handling is supported by experience with ADH receptor antagonists. However, that experience also indicates the emerging significance of autocoids, and other synergistic factors, to affect ADH receptor/effector mechanisms and to modulate renal ADH responses. PMID- 2964202 TI - Contribution of dopamine 2 receptors to dopamine-induced increase in glomerular filtration rate. AB - The effects of the interaction of dopamine (DA) and the DA2 receptors on glomerular filtration rate (GFR) has been studied by means of micropuncture technique in adult greater than or equal to 60-day-old and young 24-day-old rats. Most of the studies were performed in rats with intact adrenergic nervous systems to allow for evaluation of the presynaptic DA2 receptors. In adult and young rats, DA and LY-171555 (LY), a selective DA2 receptor agonist, induced prompt and significant increases in the single nephron (SN) GFR. Further studies were performed only in adult rats. The LY-induced increase in SNGFR was completely abolished during DA2 receptor blockade with S-sulpiride (S-SP), while the DA induced increase in SNGFR was attenuated but still significant during S-SP treatment. Both DA and LY significantly increased the glomerular ultra-filtration pressure (PUFAA). In rats with ganglionic blockade, the LY-induced increase of SNGFR was attenuated but still significant, whereas the DA-induced increase was less affected. The results imply that DA significantly increases GFR by interacting with the DA2 as well as with the DA1 receptors. Interaction between DA and the DA2 receptors increases PUFAA. The results do not rule out the existence of both pre- and postsynaptic renal DA2 receptors. PMID- 2964203 TI - Effect of atrial natriuretic factor on renal function in rats with nephrotic syndrome. AB - This study was designed to evaluate the renal effects of atrial natriuretic factor [ANF(8-33)] in rats with aminonucleoside (AMN)-induced nephrotic syndrome. AMN (100 mg/kg iv) was administered to adult female rats either 2 (AMN 2, n = 7), 4 (AMN 4, n = 7), 6 (AMN 6, n = 7), or 14 (AMN 14, n = 6) days before clearance experiments; untreated (UNT, n = 7) animals served as controls. During clearance experiments, rats were anesthetized with pentobarbital sodium. Protein excretion rates were similar between UNT and AMN 2 but increased stepwise in AMN 4, AMN 6, and AMN 14 rats. The glomerular filtration rate (GFR) was similar in UNT and AMN 2, lower in AMN 4 and AMN 14, and lowest in AMN 6 rats. Basal sodium excretion (UNaV) was not different among the five groups. An ANF primer (1.0 micrograms/kg iv) plus a constant infusion (0.1 micrograms.kg-1.min-1) for 1 h produced a significantly lower increase in UNaV in AMN 2 and AMN 14 than in UNT and was not natriuretic or diuretic in AMN 4 or AMN 6 rats. The ANF-induced increase in UNaV was similar between AMN 2 and AMN 14 rats. ANF had no effect on the GFR in any group. A higher ANF bolus (5.0 micrograms/kg iv) was then infused. This ANF bolus increased UNaV only in UNT and AMN 2 rats. Finally, a bolus of furosemide (4.0 mg/kg iv) was given; UNaV increased similarly in UNT, AMN 2, and AMN 14, and to a lesser extent in AMN 4 and AMN 6 rats. Thus, there is an attenuated natriuretic and diuretic response to ANF in rats with AMN-induced nephrotic syndrome. This altered responsiveness to ANF may contribute to the sodium and water retention characteristic of this disorder. PMID- 2964204 TI - Hemodynamic and renal effects of low-dose infusions of atrial peptide in awake dogs. AB - The effects of alpha-human atrial natriuretic peptide (alpha-hANP) on cardiovascular and renal function in conscious dogs were evaluated in two experimental protocols. In one protocol, alpha-hANP was infused intravenously at increasing rates of 50, 100, and 200 ng.min-1.kg-1 (stepup infusion) during successive 20-min periods. The greatest responses occurred during the final 20 min period of the stepup infusion when the plasma concentration of immunoreactive atrial natriuretic peptide (irANP) was increased by 44-fold over preinfusion values; pressures in the aorta and both atria were decreased at this time, whereas glomerular filtration rate, urine flow, and sodium excretion were increased. In a second protocol, alpha-hANP was infused for 1 h at constant rates of either 12.5, 25, or 50 ng.min-1.kg-1; these constant infusions increased plasma irANP by 3-, 7-, and 12-fold, respectively. Each infusion rate decreased left and right atrial pressures and increased urine flow and sodium excretion. The two lowest infusion rates elevated plasma irANP to levels that would be expected to occur only during unusual physiological, or perhaps pathophysiological, conditions. The two highest infusion rates decreased plasma renin activity. Nevertheless, the accompanying maximal increases in sodium excretion were modest (41-72%). These data imply that small changes in circulating atrial peptides that presumably occur under normal physiological conditions would not have a dominant effect on the regulation of sodium excretion; the peptides may, however, play a modulatory role on sodium excretion under these conditions. It remains to be determined whether the ability of atrial peptides to lower cardiac filling pressures is of physiological significance. PMID- 2964205 TI - Increased ANF secretion after volume expansion is preserved in rats with heart failure. AB - To examine whether the failing heart has reached a maximal capacity to increase plasma atrial natriuretic factor (ANF) concentration, the change in plasma immunoreactive ANF level due to acute blood volume expansion was determined in conscious rats with chronic heart failure. Varying degrees of myocardial infarction and thus heart failure were induced by coronary artery ligation 3 wk before study. Compared with controls, infarcted rats had decreases in mean arterial pressure (-10 mmHg, P less than 0.01), cardiac index (-27%, P less than 0.001), renal blood flow (-35%, P less than 0.01), and peak left ventricle developed pressure after aortic occlusion (an index of pressure generating ability; -15%, P less than 0.01), and increases in central venous pressure (+1.7 mmHg, P less than 0.01), left ventricular end-diastolic pressure (+10 mmHg, P less than 0.001), total peripheral resistance (+28%, P less than 0.01), and plasma ANF level (752 +/- 109 vs. 244 +/- 33 pg/ml, P less than 0.001). Plasma ANF was correlated with infarct size, cardiac filling pressures, and left ventricle pressure-generating ability. At 5 min after 25% blood volume expansion, plasma ANF in rats with heart failure increased by 2,281 +/- 345 pg/ml; the magnitude of the changes in circulating ANF and hemodynamic measurements was similar in controls. The results suggest that plasma ANF level can be used as a reliable index of the severity of heart failure, and that the capacity to increase plasma ANF concentration after acute volume expansion is preserved in rats with heart failure. There was no evidence of a relative deficiency of circulating ANF in this model of heart failure. PMID- 2964206 TI - Atrial natriuretic factor during head-out immersion at night. AB - The present investigation was undertaken to test the hypothesis that a circadian variation in the atrial natriuretic factor (ANF) response is responsible for the nocturnal inhibition of renal responses to head-out immersion (HOI). Plasma ANF responses to a 3-h HOI (water temperature 34.5 +/- 0.5 degrees C) were studied during day (1000-1300) and night (2400-0300) in six hydropenic male human subjects. In agreement with the previous observations, the renal responses, especially the diuresis, to HOI were attenuated at night compared with the day; furthermore, plasma renin activity decreased to the same low level during HOI at both day and night. Plasma ANF during time control periods was 30-40 pg/ml without showing any circadian variation. Moreover, plasma ANF showed a similar twofold increase within 1 h of HOI and was maintained at this elevated level throughout the 3-h HOI period in both the daytime and the nighttime series. On termination of HOI, plasma ANF decreased linearly to the pre-HOI level within 1 h. Hematocrit during time control periods was higher during the day compared with the night (P less than 0.05). Although HOI appears to induce a transient increase in plasma volume (as indicated by decreases in hematocrit) during the 1 h of HOI, the magnitude of the decrease in the latter parameters was not different between day and night. It is concluded that nocturnal inhibition of renal responses to HOI cannot be fully accounted for by circadian differences in the ANF and fluid shift response to HOI. PMID- 2964207 TI - Stretch-induced reduction in atrial content of natriuretic factor is locally mediated. AB - Two groups of male Wistar rats were chronically implanted with small inflatable balloons at the right superior vena caval (SVC)-atrial junction. The balloons were inflated for 60 min in one-third of the rats in each group; the remainder served as noninflated controls. Extract was prepared from the right atria of the first and from the left atria of the second group and bioassayed in male Wistars to determine the content of atrial natriuretic factor (ANF). The bioassay rats received either two injections of sham extract (control) or an initial injection (I1) of sham extract followed by an injection (I2) of balloon-inflated extract (experimental). For right atrial extract, the incremental response to the experimental injection (I2/I1) was significantly less than the response to the control injection (experimental, 1.15 +/- 0.09 mueq/min, n = 14; control, 1.59 +/ 0.15 mueq/min, n = 13; P less than 0.01). However, in the case of left atrial extract, there was no such difference between the control group (1.35 +/- 0.13 mueq/min, n = 19) and the experimental group (1.25 +/- 0.11 mueq/min, n = 11; Student's t test). We conclude that right atrial distension causes release of ANF from the right atrium, but not the left, and that this release is probably locally mediated. PMID- 2964208 TI - Thromboxane receptor-mediated bronchial and hemodynamic responses in ovine endotoxemia. AB - The role of thromboxane A2 in sheep endotoxemia, an animal model of the adult respiratory distress syndrome, was investigated by a combined biochemical and pharmacological approach. Endogenous thromboxane biosynthesis was assessed by gas chromatographic-mass spectrometric analysis of urinary (thromboxane B2, 2,3-dinor thromboxane B2) and plasma (11-dehydrothromboxane B2) metabolites that demonstrated a significant stimulation by endotoxin. The functional relevance of thromboxane A2 was probed with a specific thromboxane-prostaglandin endoperoxide receptor antagonist, SQ 29548. The antagonist significantly blunted the increase in pulmonary arterial pressure, pulmonary vascular resistance, lung lymph flow, and lymph protein clearance induced by endotoxin. Whereas the reduction in lung compliance caused by endotoxin was abolished, the augmented airway resistance was unaffected. From the simultaneous increase in thromboxane biosynthesis and effects of receptor blockade, it was concluded that thromboxane A2 mediates the early pathophysiological changes of sheep endotoxemia. Thromboxane receptor antagonism may offer a potential therapeutic approach to patients at risk of the adult respiratory distress syndrome. PMID- 2964209 TI - [Treatment of the peritoneal form of infertility using surgical laparoscopy]. PMID- 2964210 TI - Respiratory arrest after sufentanil. PMID- 2964211 TI - Allergy to propofol? PMID- 2964212 TI - Postoperative effects of intrathecal morphine in coronary artery bypass surgery. AB - To determine whether intrathecal morphine is effective in decreasing analgesic and antihypertensive drug requirements after coronary artery bypass (CAB) surgery, a prospective, randomized, double-blind study was designed. Approximately 30 minutes before induction of anesthesia with IV sufentanil and diazepam, and 2 hours before heparinization, one group of patients (n = 16) were given morphine 0.5 mg, while the control group (n = 14) were given placebo intrathecal injections through 22- or 25-gauge lumbar puncture needles. Intraoperatively, there were no differences in the numbers of patients requiring vasodilator drugs or volatile agent titration. During the postoperative period, the treated group required significantly less (P less than 0.05) IV morphine compared with the placebo group, during the first 24 hours (1.8 +/- 0.7 vs 5.4 +/ 1.5 mg) and 30 hours (2.4 +/- 0.8 vs 8.3 +/- 1.9 mg). The treated group also required significantly less (P less than 0.05) sodium nitroprusside in the first 24 hours (58.1 +/- 29.0 vs 89.1 +/- 18.4 mg). There were no differences in pain scores, and the only complications (itching, nausea and vomiting) were infrequent. It is concluded that an intrathecal dose of 0.5 mg of morphine is efficacious in reducing analgesic and antihypertensive drug requirements after CAB surgery. Whether these results are clinically important enough to warrant the theoretical risks of postheparinization lumbar hematoma is a topic for further investigation. PMID- 2964213 TI - Pharmacokinetics of sufentanil in adolescent patients with chronic renal failure. AB - The role of the kidney in sufentanil elimination or metabolism has not been defined. The effects of chronic renal failure (CRF) on the pharmacokinetic profile of sufentanil were evaluated in six adolescent patients undergoing renal transplantation, and these findings were compared with data from age-matched control patients with normal renal function who were undergoing other surgical procedures. Patients with CRF weighed significantly less than did the control patients (28.7 +/- 5.7 vs 44.7 +/- 12.9 kg [mean +/- SD]). Although there was no statistical difference in the rate of clearance or apparent volume of distribution and half-life between the two groups, clearance and half-life were more variable among patients with CRF. In these patients, therefore, sufentanil dose must be carefully administered based on responses elicited in individual patients. PMID- 2964214 TI - Two instances of seizure-like activity in the same patient associated with two different narcotics. PMID- 2964215 TI - A randomized double-blind pilot study to compare conscious sedation produced by diazepam against sufentanil. AB - Intravenous sufentanil, an analog of fentanyl, was compared to diazepam for conscious sedation in ambulatory dental outpatients. Ten patients undergoing the surgical removal of impacted third molars served as subjects in a double-blind, within-subject, single crossover study. Sedation was achieved with a combination of 30% nitrous oxide/70% oxygen by nasal mask and either diazepam (0.05-0.15 mg/kg) or sufentanic (0.05-0.15 MUg/kg) titrated to a clinical endpoint of altered speech and relaxation. Intraoperative physiologic monitoring, patients' and the oral surgeon's subjective estimates of efficacy and psychomotor recovery were used to compare the two treatments. Both patients (eight of 10) and surgeons (six of 10) preferred sufentanil sedation. No significant differences were noted between treatments for psychomotor recovery. These preliminary data in a small sample suggest that sufentanil produces adequate conscious sedation in dental outpatients and should be evaluated further with larger patient samples. PMID- 2964216 TI - Functional significance of anti-T-lymphocyte antibodies in sarcoidosis. AB - Pulmonary sarcoidosis is a chronic disorder characterized by the activation of helper/inducer T-cells in the lung without a concomitant increase in suppressor/cytotoxic T-cells. It is known that patients with sarcoidosis have circulating anti-T-cell antibodies, primarily of the IgM class. To evaluate a functional role for these antibodies in enhancing lung helper T-cell processes in pulmonary sarcoidosis, we evaluated serum and lavage fluid of patients with active sarcoidosis for the presence of anti-T-cell antibodies, the T-cell subset specificity of these antibodies, and the possible stimulatory or inhibitory effects of these antibodies on T-cells relevant to the exaggerated helper T-cell processes in sarcoidosis. Indirect immunofluorescence studies demonstrated that sarcoid patients had anti-T-cell antibodies of the IgM type reacting with autologous as well as with nonautologous normal T-cells. IgM recovered in sarcoid lavage fluid also reacted with T-cells, thus demonstrating the autoantibodies at the site of disease. Two-color immunofluorescence and flow cytometry showed that these sarcoid autoantibodies bound to mostly Leu2+ suppressor/cytotoxic T-cells, but also to a small proportion of Leu3+ helper/inducer T-cells. Incubating lymphocytes with sarcoid serum or IgM purified from sarcoid serum did not stimulate T-cell proliferation. Furthermore, when Leu2+ T-cells were stimulated with irradiated allogenic B-cells, increasing concentrations of sarcoid serum had no inhibitory effects on the activation and proliferative response of the Leu2+ T cells. Likewise, the purified IgM anti-T-cell antibodies had no inhibitory effects on the mitogenic response of Leu2+ T-cells to the anti-T-cell antigen receptor-associated T3 complex antibody OKT3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964217 TI - 31P NMR studies of the kinetics and regulation of oxidative phosphorylation in the intact myocardium. PMID- 2964218 TI - Arrangements and rearrangements of the human T-cell receptor gamma gene. PMID- 2964219 TI - Hyperthermia potentiates the activity of immunotoxin conjugates against common acute lymphoblastic leukaemia cells in vitro. AB - We studied the in-vitro cytotoxic effect of hyperthermia at 42 degrees C, both alone and in combination with ricin-linked immunotoxins, reactive with the common acute lymphoblastic leukaemia cell lines Reh and KM-3. Assessment of cytotoxicity was by incorporation of 3H-leucine and limiting dilutions analysis. The effect of immunotoxins alone and in combination with hyperthermia on normal human marrow progenitor cells was assessed by conventional colony forming units-granulocyte macrophage (CFU-GM) assay. We found that incubation of either of the cell lines with a mixture of the two immunotoxins, RPH-7-ricin and PHM-6-ricin, at 42 degrees C for one hour (h) potentiated the cytotoxic activity of the immunotoxins at 37 degrees C. At a concentration of 10(-8) mol/L, a 2.2-log kill was seen with KM-3 leukaemic cells at 37 degrees C and a 3.3-log kill at 42 degrees C, an increase of approximately 10 fold in cytotoxic activity. Survival of CFU-GM following treatment at 42 degrees C for one h with a similar concentration of immunotoxins was 26.2% (+/- 13.7%) (equivalent to 0.6 log kill) and 76.0% (+/- 1.83%) (0.1 log kill) when normal marrow was incubated with immunotoxins at 37 degrees C. This suggests relative sparing of normal marrow cells compared with the leukaemic cells tested as indicated by the 2.1-log kill difference (approximately 100 fold) between normal and leukaemic cells at 37 degrees C and the 2.7-log kill (approximately 500-fold) difference at 42 degrees C. We conclude that hyperthermia may have a role in addition to immunotoxins in the purging of marrow ex vivo to remove leukaemic cells. PMID- 2964220 TI - Endothelial linings in prosthetic vascular grafts. PMID- 2964221 TI - Thrombosis on endothelializable prostheses. PMID- 2964222 TI - Interactions of platelets and vessel wall in the development of restenosis after coronary angioplasty. PMID- 2964223 TI - [Idiopathic bi-auricular dilatation manifested by total cardiac failure. Apropos of a case confirmed by nuclear magnetic resonance]. AB - Idiopathic bilateral atrial dilatation is extremely rare. We are reporting a case in a 79 year-old patient, presenting a picture of total cardiac insufficiency. The positive diagnosis was established by bi-dimensional sonography and right angiography. Nuclear magnetic resonance confirmed the diagnosis and specified the size of the various cavities. From a rhythm standpoint, there was an atrial fibrillation without conduction disorders. The main factor of the cardiac insufficiency seems to be a low atrio-ventricular output, since the valvular insufficiency due to annular dilatation is only a secondary factor. The etiology is unknown, but a congenital origin seems most probable without excluding the possibility of an acquired structural disorder. PMID- 2964224 TI - The inheritance of Alzheimer's disease: a new interpretation. AB - Ninety-one families of Alzheimer patients were studied to determine the proportion of familial cases, to obtain pedigrees for the analysis of the mode of inheritance, and to look for clinical differences between the familial and the nonfamilial cases. The diagnosis was confirmed by autopsy in 26 cases. Thirty nine cases (43%) were familial, which is defined as more than one case in the family. Our interpretation of the pedigree data is that Alzheimer's disease is etiologically heterogeneous: it may be genetic or sporadic. In the familial type we think that the disease is inherited as an autosomal dominant, with a wide range of age of onset within a family. In one-third of these families the gene is not expressed until over age 70. No clinical differences were found between the familial and the sporadic groups. PMID- 2964225 TI - A40926, a new glycopeptide antibiotic with anti-Neisseria activity. AB - In the course of a search for glycopeptide antibiotics having novel biological properties, we isolated A40926. Produced by an actinomycete of the genus Actinomadura, A40926 is a complex of four main factors which contain a fatty acid as part of a glycolipid attached to the peptide backbone. Its activity was, in most respects, similar to that of other glycopeptides, such as vancomycin and teicoplanin. However, in addition to inhibiting gram-positive bacteria, A40926 was very active against Neisseria gonorrhoeae. A40926 was rapidly bactericidal for N. gonorrhoeae clinical isolates at concentrations equal to or slightly higher than the MIC. In mice, levels in serum were higher and more prolonged than those of an equivalent subcutaneous dose of teicoplanin. These properties suggest that A40926 may have potential in the therapy of gonorrhea. PMID- 2964226 TI - Acne of the fulminans type following testosterone therapy in three excessively tall boys. AB - Ulcerative acne was observed in three boys who underwent long-term treatment with high doses of testosterone for excessively tall stature. Even after withdrawal of testosterone therapy, this devastating type of acne still persisted for several months. After starting isotretinoin treatment, two cases progressed to full-blown acne fulminans with systemic manifestations. In these two cases, oral isotretinoin therapy induced multiple lesions of hyperproliferative granulation tissue resembling pyogenic granuloma. Topical steroid treatment proved to be beneficial for this adverse effect. Systemic corticosteroid treatment was administered in one case. High testosterone levels during puberty may be an important trigger mechanism of acne fulminans and may explain why this disease almost exclusively affects male adolescents. PMID- 2964227 TI - Hirschsprung's disease. Identification of risk factors for enterocolitis. AB - From 1975 to 1985, 80 infants and children were treated at a major pediatric hospital for Hirschsprung's disease, 19 (24%) of whom developed enterocolitis. In 9 neonates (18%) and 4 infants (29%) enterocolitis was present at diagnosis of Hirschsprung's disease, while 4 children acquired enterocolitis following a pull through procedure. Significant risk factors for development of Hirschsprung's associated enterocolitis (HAEC) were delay in diagnosis beyond 1 week of age and the presence of trisomy 21. HAEC did not occur more frequently in patients with long-segment aganglionosis, nor did an initial episode of HAEC confer a higher risk of recurrent enterocolitis. HAEC following a pull-through procedure was correlated with an anorectal stricture in three of four cases. Although neonates with HAEC had a low mortality rate (5%), their morbidity rate was 30% and their hospitalization was twice as long as neonates without enterocolitis. PMID- 2964228 TI - [The echocardiogram in arterial hypertension]. AB - Echocardiography seems to be the best non-invasive method for the detection of hypertensive heart disease: it shows early abnormalities of left ventricular compliance, frequently left ventricular hypertrophy and late abnormalities of myocardial contractility. These results are of paramount importance since recent epidemiological data have shown that left ventricular hypertrophy is a factor of severity and should be treated. PMID- 2964229 TI - [Seventy years of the Great October Socialist Revolution and the development of Soviet pathology]. AB - In reviewing briefly the changes that have taken place during the past decade (1977-86) in the theory and practice of pathology and, in the theory and practice of pathology and, more particularly, in the Pathology Service within the overall Soviet Health care system, it should be noted first of all that this period has been productive with regard to both the practical work of the pathology departments in hospitals and the research activities of scientific institutions. Pathology is gaining importance as a basic biomedical discipline. Considerable successes have been scored in fundamental research not only by the central scientific institutions but also by laboratories for morphologic research and departments of pathology at educational establishments in most of the Union Republics. Many of such laboratories and departments are well equipped and conduct work at a high technological and methodological level in areas of high priority. The number of interdisciplinary research projects has increased. A series of monographs has been published, devoted to various aspects of methodology on medicine, to the relationship between form and function, or to a critique of erroneous theories. Scientific cooperation with pathologists of other Socialist countries has been expanding, as has the participation of Soviet pathologists at international congresses, symposia, and other meetings. An important achievement has been the establishment of a network of pathology departments of hospitals and of pathologic service bureaus. There is little doubt that these bureaus which have so far been set up on a pilot basis, will become an important component of the Pathology Service on which this will rely for research progress and for keeping abreast of technological developments. They are also called upon to promote better services in rural areas. The construction of new buildings for hospital pathology departments has been expanded, but the rate and magnitude of this activity cannot yet be regarded as satisfactory. Over the past decade, pathology laboratories at hospitals have become important diagnostic centers indispensable for accurate diagnosis of many diseases through examination of biopsy specimens, and they are now better equipped than before, although the improvements have been less spectacular than expected. The quality of postmortem and clinicopathologic analysis has improved. A notable event was the session of the Collegium of the USSR Ministry of Health devoted to the Pathology Service development.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2964230 TI - An investigation of the microcirculation of the human tympanic membrane with laser-Doppler flowmetry. AB - The present investigation was performed to evaluate the use of laser-Doppler flowmetry as a means for measuring the blood flow of the microcirculation of the human tympanic membrane. The blood cell flux in the microvascular bed of the normal tympanic membrane was measured in healthy subjects. The laser-Doppler output signals continuously recorded showed a steady value ranging from 70 to about 120 V, as well as spontaneous oscillations (or rhythmical active vasomotion). Our findings show that the laser-Doppler instrument tested seems to be useful for evaluating blood flow changes in the microcirculation of the human tympanic membrane. PMID- 2964231 TI - The endorphin response of women to sleep loss and exercise. AB - The purpose of this investigation was to examine the combined effect of sleep loss and exercise on plasma beta-endorphin levels. Seven healthy women completed a 1-h run at approximately 80% VO2 max on a motorized treadmill after a normal night's sleep and again after 36 h of sleep deprivation. Venous blood samples were obtained before and immediately after each run. Oxygen uptake, heart rate, blood pressure, and ratings of perceived exertion were obtained during each trial. Resting beta-endorphin levels were 4.5 +/- 0.2 pmol.L-1 for the control trial and 4.7 +/- 0.7 pmol.L-1 for the sleep deprived trials. Exercise after normal sleep resulted in an average endorphin level of 19.4 +/- 6.8 pmol.L-1. Sleep loss did not significantly alter the post-exercise beta-endorphin response, averaging 19.4 +/- 8.7 pmol.L-1. Both trials resulted in similar oxygen uptake (41.8 +/- 2.3 ml.kg-1.min-1), heart rate (163 +/- 4 beats.min-1), mean blood pressure (98 +/- 3 mm Hg), and rating of perceived exertion (12.0 +/- 0.2). These results suggest that 36 h of sleep deprivation does not impose sufficient stress to alter the resting endorphin response, nor does it affect the endorphin response to high-intensity exercise. PMID- 2964232 TI - [In vitro identification of a child's father via the mother and vice versa by means of the mixed lymphocyte culture test]. PMID- 2964233 TI - [Maternal lymphocytes that are imprinted on the child's cells in primary mixed lymphocyte culture (MLC) are able to recognize paternal cells in secondary MLC]. PMID- 2964234 TI - [The usefulness of the MLC in questions of parentage]. PMID- 2964235 TI - 1-Deoxynojirimycin and related compounds inhibit glycogenolysis in the liver without affecting the concentration of phosphorylase a. AB - Administration in vivo of the alpha-glucosidase inhibitors 1-deoxynojirimycin and its derivatives BAY m 1099 (miglitol) and BAY o 1248 resulted in a dose- and time dependent decrease in the rate of hepatic glycogenolysis induced by glucagon. This represents a direct effect on the liver, since it could be reproduced on isolated hepatocytes. The amount of glucose produced by hepatocytes over a period of 10-20 min after addition of glucagon was decreased by about 70, 60 and 45% in the presence of maximally effective concentrations of BAY o 1248, deoxynojirimycin, and BAY m 1099, respectively. Half-maximal effects were observed at inhibitor concentrations between 20 and 100 microM. The concentrations of phosphorylase a and glycogen synthase a were not affected by inclusion of the alpha-glucosidase inhibitors in the hepatocyte suspensions. Thus, the antiglycogenolytic action of these compounds is not mediated by an altered activation state of the rate-limiting enzymes of glycogenolysis and of glycogen synthesis. PMID- 2964236 TI - Verapamil, diltiazem and nifedipine interactions with calmodulin stimulated (Ca2+ + Mg2+)-ATPase. AB - The functional interactions of the three prototype Ca2+ antagonists, verapamil, diltiazem and nifedipine, were examined in relation to the calmodulin regulated plasma membrane Ca2+ pump ATPase. For this we used low ionic strength derived, calmodulin depleted, human red cell ghost membranes. Exogenously added calmodulin activated basal (Ca2+ + Mg2+)-ATPase in a concentration-dependent manner. Half maximal activation by 6 nM calmodulin was antagonized by 10(-3) M verapamil and 10(-3) M diltiazem 25.1 and 12.1% respectively. The inhibition appeared to be specific for calmodulin activation since basal activity was not affected by these agents. Nifedipine had no effects on basal or calmodulin stimulated (Ca2+ + Mg2+) ATPase activity. Unlike dihydropyridine modulation of verapamil and diltiazem binding at high affinity channel sites, nifedipine in this system did not alter the inhibitory responses of verapamil and diltiazem. The calmodulin directed antagonism of the two drugs was shown to be strictly additive over a full range of calmodulin concentrations and appeared to change predominantly the Vmax and, to a lesser degree, the affinity of calmodulin for the (Ca2+ + Mg2+)-ATPase. It is concluded that this model system provides evidence for additional functional discrepancies among the various classes of Ca2+ antagonists. PMID- 2964237 TI - Acetylcholinesterase (EC 3.1.1.7), a neurotransmitter enzyme in scorpion hemolymph. AB - Acetylcholinesterase (AchE: EC 3.1.1.7) was identified and purified from the hemolymph of the scorpion Heterometrus bengalensis. The purity of the enzyme was determined by polyacrylamide gel electrophoresis (PAGE). The molecular weight of the enzyme, determined by sodium dodecyl sulfate-PAGE, was 80,000. The purified AchE hydrolysed acetylthiocholine iodide, but it did not react with butyrylthiocholine iodide. BW284C51, a specific inhibitor of AchE, strongly inhibited the enzyme. The known inhibitor (tetramonoisopropylpyrophosphortetramide) of pseudocholinesterase did not produce any inhibition of the enzyme activity. The purified AchE of scorpion hemolymph was vulnerable to high substrate concentration. The presence of Cu2+ and Ni2+ reduced the enzyme activity, whereas the metal ion, Sn2+, enhanced AchE activity. Ca2+ produced neither inhibition nor activation. (Na+, K+)-ATPase and Mg2+-ATPase activities were greatly enhanced by the purified AchE. PMID- 2964238 TI - Activation of the carcinogen, 5-hydroxymethylchrysene, to the mutagenic sulphate ester by mouse skin sulphotransferase. PMID- 2964239 TI - Cyclopiazonic acid inhibition of the Ca2+-transport ATPase in rat skeletal muscle sarcoplasmic reticulum vesicles. PMID- 2964240 TI - Increased helper inducer and decreased suppressor inducer phenotypes in the rheumatoid joint. AB - Cells isolated from the joints of patients with rheumatoid arthritis (RA) exhibit functional immune abnormalities, such as diminished suppressor activity, depressed response to mitogens, and enhanced immunoglobulin production. We sought to characterize the T lymphocyte subsets in the synovial fluid (SF) and peripheral blood (PB) of RA patients in an attempt to clarify the mechanism(s) responsible for these functional immune abnormalities. We used dual immunofluorescence staining techniques with several combinations of monoclonal antibodies, including anti-4B4 and anti-2H4, which define, respectively, the helper inducer and suppressor inducer subsets of CD4+ (Leu-3+ and T4+) cells. Mononuclear cells from normal PB (n = 9), RA PB (n = 6), and RA SF (n = 9) were analyzed, after staining, by flow cytometry. We observed a significant increase (P less than 0.0002) in the number of cells bearing the helper inducer phenotype (CD4+, 4B4+), and a significant decrease (P less than 0.0002) in the number of cells bearing the suppressor inducer phenotype (CD4+, 2H4+), in RA SF compared with the levels in PB from RA patients or normal control subjects. We also observed that the CD8+, 2H4+ subset was significantly decreased (P less than 0.0001) in SF compared with that in PB. There was no significant difference in the lymphocyte subset levels in PB from RA patients and from normal subjects. These observations may account, in part, for the reduced suppressor activity, the poor response to mitogens, and the autologous mixed lymphocyte reaction, as well as the enhanced production of Ig and rheumatoid factor, that are observed in the rheumatoid joint. PMID- 2964241 TI - Biotransformation studies of di-acid angiotensin converting enzyme inhibitors. AB - The biotransformation of di-acid inhibitors of angiotensin converting enzyme was studied in the urine of rats using gas chromatography/mass spectrometry. It was found that after oral administration (10 mg/kg) of enalapril significant amounts (9.2%) of a hydrolytic metabolite of enalaprilat were excreted in urine which was identified as 2-N-alanyl-4-phenylbutanoic acid. This metabolite was present only in trace concentrations in urine after intravenous administration. This pathway was not present, however, with either ramipril or perindopril suggesting that the amide bond in these newer inhibitors is more resistant to hydrolysis than for enalapril. Glucuronidase hydrolysis of urine obtained from rats dosed with either enalapril, ramipril or perindopril indicated the absence of glucuronidate conjugates of these inhibitors in rat urine. PMID- 2964242 TI - Effects of nicorandil on large epicardial coronary artery in conscious dogs. AB - The effect of 2-nicotinamidoethyl nitrate (nicorandil, SG-75, Sigmart) on coronary circulation was studied in conscious dogs, instrumented previously under sterile condition with sonomicrometers for the external coronary diameter measurement, and an electromagnetic flow plobe on the left circumflex coronary artery and a catheter into the thoracic aorta. Intravenous administration of nicorandil in doses of 10, 30, 100 and 300 micrograms/kg increased coronary blood flow dose-relatedly by 13 +/- 2, 24 +/- 3, 144 +/- 18 and 309 +/- 36%, respectively. Nicorandil also increased the diameter of the large epicardial coronary artery by 38 +/- 6, 71 +/- 11, 150 +/- 24 and 173 +/- 26 microns in doses of 10, 30, 100 and 300 micrograms/kg, respectively. To eliminate flow dependent dilation of the coronary artery, the external diameter of the large epicardial coronary artery was measured during fixing the amount of coronary blood flow constant by a cuff occluder after intravenous administration of nicorandil, however, the degree of coronary diameter increase was not attenuated. Thus, nicorandil dilates both large and small coronary arteries in conscious dogs, the former dilation being independent from changes in coronary blood flow. PMID- 2964243 TI - Traumatic intermuscular hernia in the anterior abdominal wall. AB - Traumatic ventral herniae are uncommon. The authors present a patient with traumatic intermuscular hernia in the anterior abdominal wall. Early recognition and differentiation from haematoma is important. In this case, the overlying intact muscle layer masked the classical clinical signs. The incision should be adequate and all internal organs carefully examined. Primary repair is usually possible. PMID- 2964244 TI - The abdominal wall test. PMID- 2964245 TI - [Prediction of growth failure in children young under a year old]. PMID- 2964246 TI - [Penicillin-resistant Neisseria gonorrhoeae in Mexico City]. PMID- 2964248 TI - Recent developments in rehabilitation giving rise to important new (and old) ethical issues and concerns. PMID- 2964247 TI - Murine cutaneous leishmaniasis: susceptibility correlates with differential expansion of helper T-cell subsets. AB - BALB/c mice develop fatal illness following infection with Leishmania major despite expansion of helper L3T4+ T cells in the draining lymph nodes and spleen. Healer mice, either genetically resistant C57BL/6 or BALB/c that have been pretreated with monoclonal antibody GK 1.5, also develop expanded numbers of L3T4+ T cells at the time of healing. Lymph node cells from healer mice produce gamma-interferon in vitro and message for gamma-interferon can be recovered from the lymph nodes during healing in vivo. Conversely, cells harvested from non healer mice during the course of infection produce minimal gamma-interferon in vitro and have little message for gamma-interferon detectable in vivo. When the same Northern blots are hybridized for IL-4, large amounts of IL-4 message are detected only in cells from non-healer mice. The data are consistent with the expansion of type 1 helper cells (Th1) during healing and type 2 helper cells (Th2) during progressive leishmania infection. PMID- 2964249 TI - Down's syndrome and maternal cancer. A preliminary study. AB - The possible association between maternal cancer and Down's syndrome was studied in a register-based case-control study of 962 women who had an infant with Down's syndrome born during 1973-1981, and to each two controls, matched for maternal age, parity, and year of delivery. A total of 952 complete triplets was obtained. Among them, 63 Down's syndrome mothers and 79 control mothers were found in a cancer registry for the period 1958-1982 (odds ratio 1.6;95% conf. interval 1.3 2.0). Of these, 14 Down's syndrome mothers and 20 controls had malignant lesions (odds ratio 1.4, 0.9-2.2), and 49 Down's syndrome mothers and 55 controls had cervical cancer in situ (odds ratio 1.8, 1.4-2.3). This risk increase was present both before and after the delivery of the Down's syndrome infant. Various explanations of these findings are discussed and the need for further data is stressed. PMID- 2964250 TI - The HIV positive patient. PMID- 2964251 TI - New data on the acceptance of the hepatitis B vaccine by dental personnel in the United Kingdom. PMID- 2964252 TI - The effect on microleakage of interchanging dentine adhesives in two composite resin systems in vitro. PMID- 2964253 TI - Once daily amlodipine in the treatment of mild to moderate hypertension. AB - 1. The antihypertensive efficacy of once-daily amlodipine was studied in a group of 30 patients with mild to moderate hypertension in a double-blind, placebo controlled parallel group study. The dose range of amlodipine was 2.5-10 mg daily titrated at 2 weekly intervals for a total treatment period of 8 weeks. 2. Amlodipine produced a significant reduction in blood pressure compared with placebo, the mean difference between baseline and 8 weeks (corrected for placebo effect) being 16/12 mm Hg supine, 14/4 mm Hg standing. 3. Blood pressure returned to baseline values during a terminal 4 week washout period on placebo. 4. There were no significant effects on heart rate. 5. Two patients experienced slight ankle oedema while receiving amlodipine 10 mg daily but the active drug was otherwise well tolerated. 6. Plasma concentration of amlodipine, sampled 24 h after the preceding dose, increased as the dose titration sequence was followed, averaging 2.5 ng ml-1 on 2.5 mg, 4.9 ng ml-1 on 5 mg and 10.5 ng ml-1 on 10 mg. PMID- 2964254 TI - T prolymphocytoid leukaemic transformation of a T chronic lymphoid leukaemia characterized by surface markers as helper T cells for B cell immunoglobulin synthesis. PMID- 2964255 TI - Selective decrease in platelet dense granule adenine nucleotides during recovery from acute experimental thrombocytopenia and ensuing thrombocytosis in baboons. AB - Serial measurements of platelet volume, platelet content of adenine nucleotides, beta-thromboglobulin (beta-TG), platelet factor 4 (PF4) and ex vivo platelet aggregation were made in baboons under basal, steady-state conditions of normal platelet production, and during recovery from acute thrombocytopenia induced by the exposure of flowing blood to spherical glass microbeads. The mean basal platelet count of 509 +/- 107 X 10(9)/l (+/- 1 SD; n = 4) fell acutely to 36.8 +/ 12.6 X 10(9)/l after the insertion of glass bead columns and blood filters, placed distally, for 60 min into surgically implanted arteriovenous-shunts in heparinized baboons. After the irreversible removal of up to 90% of the baseline circulating platelet population, recovery from thrombocytopenia was characterized by a constant rate of increase in circulating platelet counts (115 +/- 11 X 10(9)/l/d) and a rebound thrombocytosis to 1.5 times the basal platelet count after 7 d. Steadystate thrombocytopoiesis was achieved by 3-4 weeks after the onset of thrombocytopenia. Platelet dense granule ADP and ATP decreased significantly from 3.89 +/- 0.20 and 2.33 +/- 0.25 mumol/10(11) platelets respectively at baseline to 2.17 +/- 0.37 and 1.68 +/- 0.37 mumol/10(11) platelets respectively after 7 d (P less than 0.001 in both cases) and normalization was achieved only after 4 weeks. By contrast, the mean platelet volume and platelet content of beta-TG and PF4 did not change significantly throughout the course of study (P greater than 0.1 in both cases). Platelet function, assessed by platelet aggregation ex vivo, demonstrated that platelet function was not impaired despite the significant decrease in dense granule ADP. We conclude that a selective, temporal reduction in platelet dense granule adenine nucleotides reflects changes in the thrombocytopoietic control mechanism secondary to induction of acute thrombocytopenia. PMID- 2964256 TI - The effect of electroconvulsive therapy on endorphins in depression. AB - The time course study of the endorphin response to electroconvulsive therapy (ECT) was carried out in 10 patients (including one control who did not receive active ECT) at the first and sixth ECT. Results showed a significant rise of plasma endorphin levels after ECT. This increase returned to the pre-ECT level within 1 hr after ECT. There was a pre-rise of plasma endorphin level, which probably was stress related and which was also observed in the control case. PMID- 2964257 TI - The dependence of isometric tension, isometric ATPase activity, and shortening velocity of limulus muscle on the MgATP concentration. AB - The dependence of the isometric tension, the velocity of unloaded shortening, and the steady-state rate of MgATP hydrolysis on the MgATP concentration (range 0.01 5 mM MgATP) was studied in Ca-activated skinned Limulus muscle fibers. With increasing MgATP concentration the isometric tension increased to a peak at approximately 0.1 mM, and slightly decreased in the range up to 5 mM MgATP. The velocity of unloaded shortening depended on the MgATP concentration roughly according to the Michaelis-Menten law of saturation kinetics with a Michaelis Menten constant Kv = 95 microM and a maximum shortening velocity of 0.07 muscle lengths s-1; the detachment rate of the cross-bridges during unloaded shortening was 24 s-1. The rate of MgATP splitting also depended hyperbolically on the MgATP concentration with a Michaelis-Menten constant Ka = 129 microM and a maximum turnover frequency of 0.5-1 s-1. The results are discussed in terms of a cross bridge model based on a biochemical scheme of ATP hydrolysis by actin and myosin in solution. PMID- 2964259 TI - Low total protein S antigen but high protein S activity due to decreased C4b binding protein in neonates. AB - Protein S, a vitamin K-dependent cofactor for activated protein C, exists in normal adult plasma in a free anticoagulantly active form and in an inactive form complexed to C4b-binding protein. Immunologic and functional levels of protein S and C4b-binding protein in plasma were determined for 20 newborn infants and compared with adult normal pooled plasma. Total protein S antigen levels averaged 23%, similar to other vitamin K-dependent plasma proteins. However, the protein S anticoagulant activity was 74% of that of adult normal plasma. This apparent discrepancy of activity to antigen was shown to be due to low or undetectable levels of C4b-binding protein, which results in the presence of most if not all of protein S in its free and active form. The relatively high level of anticoagulantly active protein S in infants may enhance the potential of the protein C pathway, thereby minimizing risks of venous thrombosis in this group. PMID- 2964258 TI - A fluorescence photobleaching study of the microsecond reorientational motions of DNA. AB - We have conducted a polarized fluorescence photobleaching recovery (FPR) study of the rotational dynamics of ethidium azide labeled DNA. Polarized photobleaching experiments provide data on microsecond and millisecond molecular reorientation that complement the information available from nanosecond fluorescence depolarization studies. In polarized FPR experiments an anisotropic angular concentration of fluorophore is created by bleaching dye molecules in a preferred orientation with a short, intense pulse of polarized light. The sample is then weakly illuminated, and the temporal variation in the emitted fluorescence is monitored. The fluorescence signal will systematically change as molecules undergo post-bleach reorientation and the angular distribution of dye tends toward isotropy. We have observed that the time dependence of our microsecond FPR curves is also determined in part by nonrotational phenomena. To isolate the reorientational recovery we conduct our FPR experiments in two modes (called parallel and perpendicular) that differ only in the polarization of the bleaching light. A quotient function, R(t), is constructed from the data obtained in these two modes; the variation with time of this new quantity is governed solely by processes that are sensitive to the polarization of the incident light (e.g., molecular rotation). It is found experimentally that R(t) remains constant, as expected, for rotationally restricted DNA systems despite a temporal recovery in the parallel and perpendicular FPR curves. We also follow the dynamics of solutions of phage lambda DNA as revealed in the temporal dependence of R(t). This DNA system rotationally relaxes after approximately 100 microseconds and the dye/DNA complex reorients substantially during the 10-microseconds bleach period. Our FPR data are interpreted in terms of dynamic models of DNA motion. PMID- 2964260 TI - Cost of limiting abortion. PMID- 2964261 TI - Noradrenaline action on cat retinal ganglion cells is mediated by dopamine (D2) receptors. AB - Effects of iontophoretically applied noradrenaline, dopamine and their receptor antagonists on the retinal ganglion cells, were studied in optically intact eyes of barbiturate-anaesthetized cats. Noradrenaline inhibited visually evoked and spontaneous firing of all classes of retinal ganglion cells: the effect being greater on ON- than on OFF-cells and slightly more potent than dopamine on a given cell. All alpha- and beta-adrenoreceptor blockers tested tended to change spikes, but were generally ineffective in blocking the noradrenaline-induced inhibition, when not affecting spikes. The noradrenaline-induced inhibition was, however, effectively blocked by dopamine D2-receptor antagonists. The alpha- and beta-adrenoreceptor antagonists applied alone had no effect, suggesting the absence of endogenous noradrenergic antagonism, although alpha-type adrenergic antagonism was suggestive on a very small number of cells. These results suggest that: (1) noradrenaline action on cat retinal ganglion cells is mediated via dopamine D2-receptors; (2) noradrenaline is not generally released on them, except there may be physiologically active alpha-receptors on a few cells; and (3) many of the adrenoreceptor blockers affect membrane properties of the retinal ganglion cells, in a similar manner to local anaesthetics. PMID- 2964262 TI - [Morphologic characteristics of isolated hypertrophic cardiomyocytes in scanning electron microscopy]. PMID- 2964263 TI - A survey of a program for the dental care of disabled adults. PMID- 2964264 TI - Induction of progesterone receptor with tamoxifen in human breast cancer with special reference to its behavior over time. AB - The behavior of progesterone receptor (PR) values over time in human breast cancer during tamoxifen treatment was studied. Of 97 patients with operable breast cancer, 24 were preoperatively treated with tamoxifen (20 mg twice daily) for 3 days (TAM 3-day), 22 were treated for 7 days (TAM 7-day), 21 for 14 days (TAM 14-day), and 30 received no treatment (control group). Surgically removed breast tumors were assayed for the progesterone receptor by the dextran-coated charcoal method. The PR values (fmol/mg DNA) (mean +/- standard error [SE]) of PR positive tumors in the control, TAM-3, TAM-7, and TAM-14 day groups were 571 +/- 176, 1699 +/- 408 (P less than 0.01), 1675 +/- 463 (P less than 0.05), and 686 +/ 191, respectively. There was a 4.6-fold increase in the mean PR value in postmenopausal patients and a two-fold increase in premenopausal patients after TAM treatment for 3 and 7 days. We concluded that PR induction provoked by TAM reached a peak on day 3, continuing at this level until day 7. A longer treatment (14 days) with TAM appears to abolish the PR inducing activity, probably because of, first, estrogenic and, later, the antiestrogenic effects of TAM. PMID- 2964265 TI - Fragile sites and genitourinary tumors. AB - We tested for fragile sites in lymphocytes from nine patients with genitourinary tumors to determine if a correlation existed between their cancer chromosome breakpoints and fragile sites. Induction was done for rare fragile sites in all known classes by exposure of cells to fluorodeoxyuridine and bromodeoxyuridine (BrdU). No rare fragile sites were found. Induction was also done for common fragile sites in all known classes using aphidicolin (Apc), 5-azacytidine, and BrdU. Although 56 common fragile sites were detected, only a single site corresponded in location to a genitourinary tumor chromosome breakpoint. That was the common fragile site in band 3p14. No overall correlation was found between fragile sites and chromosome rearrangements in carcinoma of the kidney, ureter, bladder, and testis. The sole known candidate for a possible biologic role is the 3p14 common fragile site in renal cell carcinoma. PMID- 2964266 TI - Phenotype of syngeneic tumor-specific cytotoxic T-lymphocytes and requirements for their in vitro generation from tumor-bearing host and immune spleens. AB - Cells required for the in vitro generation of syngeneic cytotoxic T-lymphocytes (CTL) against the P815 mastocytoma in the DBA/2 mouse strain were investigated. For both immune and tumor-bearing host spleen cells, CTL effector cells were eliminated by treatment with anti-Thy1.2, anti-Lyt1.1, or anti-Lyt2.1 and C', but were resistant to anti-L3T4 (GK1.5). Thus, CTL effectors (and their precursors) were Lyt1+2+, L3T4-. However, P815-specific CTL could not be generated in the absence of L3T4+ cells, whose function could be replaced with exogenous interleukin-2 (IL-2). When monoclonal antibodies against L3T4 were added to mixed leukocyte tumor cultures, CTL generation was markedly inhibited. Depletion of accessory cells also led to a marked reduction in CTL generation, which could be restored to control levels by adding adherent cells from normal spleens or with exogenous IL-2, but not with IL-1. Thus, accessory cells are apparently required to present the tumor antigens of this Ia-negative tumor to T-helper cells. PMID- 2964267 TI - Antiemetic properties of the 5HT3-receptor antagonist, GR38032F. AB - GR38032F is a highly selective 5HT3-receptor antagonist which inhibits vomiting induced by cisplatin, cyclophosphamide or X-radiation in the ferret. Since cisplatin selectively increased the levels of 5HT and 5HIAA in the intestinal mucosa, a possible site of the antiemetic action of GR38032F may be on 5HT3 receptors on vagal afferents in the small intestine. The potent antiemetic action of GR38032F should be of clinical value in reducing the nausea and vomiting associated with radiotherapy or chemotherapy of cancer. PMID- 2964268 TI - The augmentation of tumor-specific immunity using haptenic muramyl dipeptide (MDP) derivatives. III. Eradication of disseminated murine chronic leukemia cells by utilizing MDP hapten-reactive helper T-cell activity. AB - A previous paper has demonstrated that enhanced tumor-specific immunity could be induced by priming mice with Bacillus Calmette Guerin (BCG) and subsequently immunizing them with syngeneic tumor cells modified with BCG-cross-reactive muramyl dipeptide (MDP) hapten. The present study establishes a tumor-specific immunotherapy protocol for a murine chronic leukemia based on the above T-T cell collaboration between antitumor effector T cells and anti-MDP hapten helper T cells induced by BCG priming. BALB/c mice which had been primed to BCG were injected intravenously (i.v.) with viable, syngeneic BCL1 leukemia cells. One week later, these mice were immunized intraperitoneally (i.p.) with unmodified or MDP hapten-modified, 10,000 R X-irradiated BCL1 cells, followed by 4 booster immunizations at 5-day intervals. The administration of unmodified BCL1 tumor cells into BCG-primed mice failed to prevent them from tumor death due to the persistent growth of preinjected BCL1 cells. In contrast, the immunization of BCG primed, BCL1 leukemia-cell-bearing mice with MDP-modified BCL1 cells resulted in a high growth inhibition of leukemia cells and protection of these mice from death by leukemia. It was also revealed that potent tumor-specific, T-cell mediated immunity was generated in mice which survived in this immunotherapy model. Thus, these results indicate that administration of MDP hapten-modified, syngeneic leukemia cells into leukemia-bearing mice which have been primed with BCG results in potent tumor-specific, T-cell-mediated immunity attributable to preventing the growth of disseminated leukemic cells. PMID- 2964269 TI - Sublethal, whole-body ionizing irradiation can be tumor promotive or tumor destructive depending on the stage of development of underlying antitumor immunity. AB - It was shown that sublethal (500 rads), whole-body gamma-irradiation of mice bearing an established i.d. immunogenic tumor can result, after several days delay, in complete tumor regression and long-term survival, but only if radiation is given after the tumor is established and growing progressively. Exposing mice to the same dose of radiation several hours after tumor cells were implanted resulted, in contrast, in enhanced growth of the primary tumor and in earlier death from systemic disease. Irradiation-induced tumor regression failed to occur in mice that were incapable of generating antitumor immunity, because of having been made T cell deficient by thymectomy and irradiation. Again, irradiation induced tumor regression could be blocked by infusion of spleen cells from donor mice bearing a well-established tumor. These and previously published results support the view that sublethal, whole-body ionizing irradiation causes tumor regression by preferentially destroying radiosensitive suppressor T cells, thereby enabling the host to generate a therapeutic level of concomitant immunity. It is suggested that the preferential destruction of suppressor cells by irradiation depends on the acquisition, during immunologic induction, of radioresistance by antigen-activated effector T cells, and that this is the reason irradiation causes regression only of established tumors. Not all tumors tested were immunogenic enough to undergo regression in response to gamma irradiation. PMID- 2964271 TI - Paget's disease: state of the art. PMID- 2964270 TI - Response of natural killer cells from dietary tyrosine- and phenylalanine restricted mice to biological response modifiers. AB - The effect of dietary tyrosine and phenylalanine restriction on splenic natural killer (NK) cell activity was studied in tumor-free B6D2F1 and NIH nude mice and in B16 bladder-6 (BL6) melanoma-bearing B6D2F1 mice. This dietary restriction was found to suppress the naturally elevated NK-cell activity of nude mice and to induce a specific lymphocytopenia in B6D2F1 mice fed the restricted diet for a prolonged period. Baseline NK-cell activity was significantly lower in tumor-free B6D2F1 mice fed a diet restricted in tyrosine and phenylalanine (restricted diet) than in tumor-free mice fed a basal diet. Similar kinetics of activation after a single i.p. injection of 100 micrograms of polyinosinic:polycytidylic acid (poly I:C) were observed in mice fed both diets. NK-cell activity was not significantly augmented after i.v. inoculation of BL6 melanoma, irrespective of the diet fed; however, it was enhanced in tumor-bearing mice after poly I:C injection. This augmentation was similar to that observed in tumor-free mice. Spleen cells from mice fed either diet were responsive to stimulation of NK-cell activity after in vitro incubation with interleukin-2. These results indicate that dietary restriction of tyrosine and phenylalanine, a potentially useful therapeutic adjunct known to lower NK-cell activity, does not significantly interfere with poly I:C or interleukin-2 induction of NK cells. Our results also demonstrate that, while this dietary restriction causes lymphocytopenia, no effect of the diet could be found on total serum IgG or circulating immune complex levels. PMID- 2964272 TI - Differential expressions of phosphofructokinase isozymes in different tissues of young and old rats. PMID- 2964273 TI - Structural diversity in muscle fibres of chicken breast. AB - Chicken breast muscle is usually considered to be a relatively homogeneous white muscle and has therefore been widely used for studies of muscle proteins. In a previous study, however, we have found different M-region structures in different fibres from this muscle. Because of this result, we have now carried out a combined histochemical and ultrastructural survey of this muscle. In particular, we have made use of large transverse cryo-sections that include most of the muscle cross-section. Although the white region is fairly homogeneous in fibre content according to normal histochemical criteria (mATPase), we have found that there is a gradation of fibre structure across the muscle. The bulk of the muscle stains conventionally for Type-II fibres according to mATPase tests (the "white" part) but, in the small "red" part of the muscle, there are also Type-I fibres together with the Type-II fibres. Superimposed on this division into Type-I and Type-II fibres are variations in fibre size, oxidative and glycolytic staining properties, and variations of Z-band width and M-band structure; there is no strict correlation among any of these parameters. The apparently uniform staining across most of the muscle when tested for myofibrillar ATPase may be a misleading indicator of fibre properties. PMID- 2964274 TI - Synapsis of attachment sites during lambda integrative recombination involves capture of a naked DNA by a protein-DNA complex. AB - During lambda integration, Int recombinase must specifically bind to and cut attachment sites on both the viral and host chromosomes. We show here by foot printing and by a novel cleavage assay that the bacterial attachment site, attB, cannot stably bind Int in competition with other DNAs. Instead, during recombination reactions, attB obtains its Int by collision with the intasome, a nucleoprotein assembly that forms on the viral attachment site, attP. Our cleavage assay also shows that the capture of attB by the attP intasome does not depend on DNA homology between the two sites; synapsis is governed solely by protein-protein and protein-DNA interactions. PMID- 2964275 TI - Alzheimer's disease: its proteins and genes. PMID- 2964276 TI - The mannose 6-phosphate receptor and the biogenesis of lysosomes. AB - Localization of the 215 kd mannose 6-phosphate receptor (MPR) was studied in normal rat kidney cells. Low levels of receptor were detected in the trans Golgi network, Golgi stack, plasma membrane, and peripheral endosomes. The bulk of the receptor was localized to an acidic, reticular-vesicular structure adjacent to the Golgi complex. The structure also labeled with antibodies to lysosomal enzymes and a lysosomal membrane glycoprotein (lgp120). While lysosome-like, this structure is not a typical lysosome that is devoid of MPRs. The endocytic marker alpha 2 macroglobulin-gold entered the structure at 37 degrees C, but not at 20 degrees C. With prolonged chase, most of the marker was transported from the structure into lysosomes. We propose that the MPR/lgp-enriched structure is a specialized endosome (prelysosome) that serves as an intermediate compartment into which endocytic vesicles discharge their contents, and where lysosomal enzymes are released from the MPR and packaged along with newly synthesized lysosomal glycoproteins into lysosomes. PMID- 2964277 TI - Molecular cloning of an enhancer binding protein: isolation by screening of an expression library with a recognition site DNA. AB - A novel strategy has been used to isolate a cDNA clone that encodes a DNA binding domain whose recognition properties overlap those of the mammalian transcription factors H2TF1 and NF-kappa B. These two factors are distinguished by their cell type distributions and their relative affinities for related sequence elements in the enhancers of the major histocompatibility complex (MHC) class I and immunoglobulin kappa chain genes. The human cDNA clone was detected by screening a lambda phage expression library with a binding site probe derived from the MHC enhancer. The phage encoded fusion protein binds specifically to both the MHC and kappa gene enhancers. The cDNA hybridizes to a single copy gene that is expressed as a 10 kb mRNA in both B and non-B cells. The strategy used in this study may prove generally useful in the cloning and analysis of sequence-specific DNA binding proteins. PMID- 2964278 TI - Rotation and translocation of microtubules in vitro induced by dyneins from Tetrahymena cilia. AB - Dynein, the force-generating enzyme that powers the movement of cilia and flagella, has been characterized biochemically, but no simple system has been available for examining its motile properties. Here we describe a quantitative in vitro motility assay in which dynein adsorbed onto a glass surface induces linear translocation of purified bovine microtubules. Using this assay, we show that both 22S and 14S dyneins from Tetrahymena cilia induce movement but have distinct motile properties. A unique property of 14S dynein, which has not been described for other motility proteins, is its ability to generate torque that causes microtubules to rotate during forward translocation. In the axoneme, 14S dynein induced torque may induce rotation of central-pair microtubules and may play an important role in generating three-dimensional ciliary beating patterns. PMID- 2964279 TI - Characterization of an endogenous Lyt2+ T-suppressor-cell population regulating autoreactive T cells in vitro and in vivo. AB - Autoreactive T cells have been defined by their capacity to respond to self-Ia antigens expressed on non-T cells. Several recent studies have suggested that these cells may play important immunoregulatory functions. However, it is not clear what regulates the responsiveness of autoreactive T cells and why such cells are not demonstrably stimulated in vivo, where they are in the constant presence of self-Ia antigens. In the present study we examined the role of T suppressor (Ts) cells in regulating autoreactive T cells. We observed that enhanced autoreactivity occurred in vitro when Lyt2+ T cells were depleted from the responding and/or stimulating spleen cells in a syngeneic mixed-lymphocyte reaction. Similarly, addition of irradiated Lyt2+ T cells but not L3T4+ T cells inhibited the response of L3T4+ T cells to self-Ia antigens. The activity of the suppressor cells was specific to the autoreactive T cells since antigen-specific and alloreactive T-cell proliferation were not inhibited. Furthermore, depletion of Lyt2+ T cells by in vivo treatment of mice with anti-Lyt2 monoclonal antibodies caused enhanced endogenous proliferation of lymph node and splenic T cells and increased the T-cell response to self-Ia antigens in vitro. These studies, therefore, suggest that T-cell tolerance to self-Ia antigens in vivo may be maintained by naturally occurring Lyt2+ Ts. Mice having enhanced autoreactivity may provide a useful tool to address the role of autoreactive T cells in the immune response to foreign antigens and in the pathogenesis of autoimmune diseases. PMID- 2964280 TI - [Con A-induced suppressor activity in pulmonary tuberculosis]. PMID- 2964281 TI - [Effect of cardionatrin on experimental respiratory distress syndrome by oleic acid in Wistar rats]. PMID- 2964282 TI - Calmodulin involvement in TPA and DDT induced inhibition of intercellular communication. AB - The organochlorine pesticide DDT is a liver tumour promoter and a potent inhibitor of intercellular communication. Present knowledge of the mechanism by which DDT inhibits intercellular communication is limited but it has been suggested that increased intracellular free calcium induced by DDT could be of importance. As the effects of calcium are closely associated with the multifunctional protein calmodulin (CaM) in most cells the potential binding of DDT to CaM and subsequent effects on CaM-stimulated Ca2+/Mg2+-ATPase activity were studied. DDT inhibited CaM-stimulated Ca2+/Mg2+-ATPase activity and bound to CaM in a manner similar to established CaM-inhibitors. Subsequently an in vitro assay for measuring inhibition of metabolic cooperation between 6-thioguanine (TG)-sensitive and TG-resistant Chinese hamster (V79) cells was used to investigate the possible involvement of CaM in the regulation of intercellular communication. Calmidazolium (CzM), a potent CaM inhibitor, was tested alone or in combination with the tumour promoters 12-O-tetradecanoyl phorbol-13-acetate (TPA) or DDT known inhibitors of intercellular communication. The results showed that CzM alone was without effect with regard to inhibition of metabolic cooperation but potentiated the response induced by TPA, an effect not noticed with DDT. These results suggest different mechanisms of action of TPA and DDT on metabolic cooperation and support the hypothesis that with calcium CaM may be of importance for drug-induced inhibition of intercellular communication and tumour promotion. PMID- 2964283 TI - The relationship between lipophilic-hydrophilic balance, uptake and anti bacteriophage lambda activity of experimental anti-tumour bisquaternary salts. AB - The uptake by Escherichia coli of a series of bisquaternary experimental anti tumour agents (quinolinium 4-[p-9(-pyridylamino)phenylcarbamoyl]-aniline-bisalkyl dibromides) has been measured both by association of radiolabelled compounds and their inhibition of the vegetative replication of bacteriophage lambda (after heat inactivation of the phage repressor) as a measure of biologically effective intracellular drug concentration. Uptake of these compounds was correlated with biological effect, and was a function of both incubation temperature and the lipophilic-hydrophilic balance of the compound. At 30 degrees C uptake was drug concentration-dependent and was not readily reversible. No saturation of uptake was apparent over the concentration range tested. Preliminary experiments indicated that time-dependent drug uptake was also related to growth inhibition in cultured L1210 murine leukaemia cells. These results are consistent with the hypothesis that uptake occurs by diffusion across the plasma membrane followed by strong binding to cell constituents such as DNA. The approximate range of uptake of the most active compounds, using an external drug concentration of 1 microM, are 100 and 2400 molecules/s respectively for bacteria and murine leukemia cells. For bacteria, the uptake of approx. 2 X 10(5) molecules of drug/cell inhibits the yield of phage lambda by 90%. PMID- 2964284 TI - [Comparison between the sinigrin content of the Sinapis (Brassica juncea) before and after processing]. PMID- 2964285 TI - [Authentic species of the Chinese herbal drug qinghao]. PMID- 2964286 TI - [Determination of the content and release in vitro of the suppository of Filifolium sibiricum]. PMID- 2964287 TI - [Analysis of nutrient constituents in Dioscorea opposita]. PMID- 2964288 TI - [Chemical composition of the volatile oil from Eupolyhaga sinensis]. PMID- 2964289 TI - [HPLC identification for guanxinsuhe pills]. PMID- 2964290 TI - [Direct gas chromatography determination of amygdalin in Armeniacae seeds by enzymolysis]. PMID- 2964291 TI - [Inhibitory action of sophocarpine on the central nervous system]. PMID- 2964292 TI - [Pharmacological studies on the neutral oil of gao ben (Ligusticum sinense) (II): The inhibitory effect on the smooth muscle of the intestines and uterus]. PMID- 2964293 TI - [Anti-snake bite action of Picrasma quassioides]. PMID- 2964294 TI - [Be on the alert against the incompatibility of Chinese traditional drugs with Occidental medicines]. PMID- 2964295 TI - [Revisions of some botanical names in the Chinese Pharmacopoeia (1985 edition) Part I]. PMID- 2964296 TI - [Identification of fructus Arctii and fructus Onopordon acanthii]. PMID- 2964297 TI - [Suggestions on some Latin names and terms in the newly revised textbook, "Pharmaceutical Botany"]. PMID- 2964298 TI - [UV-spectrophotometry of total flavonoids in maidong]. PMID- 2964299 TI - [High performance liquid chromotographic determination of two tanshinones and the chromotographic profiles of 15 danshens]. PMID- 2964300 TI - [Growing trend and changes in saponin content of the root of Panax quinquefolium at different developmental stages]. PMID- 2964301 TI - [Research on the traditional process of preparing realgar]. PMID- 2964302 TI - [A design of the factory for culturing commercial glycyrrhiza to be built in East North China]. PMID- 2964303 TI - [Influence of extraction methods on the anthraquinone content of rhubarb]. PMID- 2964304 TI - [Determination of lead in the urine of patients using medicated plaster]. PMID- 2964305 TI - [Chemical constituents of Rabdosia japonica var. glaucocalyx]. PMID- 2964306 TI - [Comparison of the constituents of the rhizome Corydalis (Corydalis yanhusuo) cultured in Deng County, Honan Province and that cultured in Zhejiang Province]. PMID- 2964307 TI - [Chemical constituents of fructus Lycii and folium Lycii (II): amino acid in fructus Lycii and folium Lycii]. PMID- 2964308 TI - [Quantitative determination of arbutin and gallotannin in Pyrola calliantha]. PMID- 2964309 TI - [Preliminary research on the pharmacological effect of the oral liquid medicine guanxinan]. PMID- 2964310 TI - [Some pharmacologic actions of the combination of jiangbanxia with zhifuzi]. PMID- 2964311 TI - [Treatment of acute tonsillitis with total organic acid capsules of Achillea]. PMID- 2964312 TI - [Studies on the literature on the rhizome renshen]. PMID- 2964313 TI - [Textual studies on the snow silkworm]. PMID- 2964314 TI - [Pharmacognosy study of the Chinese drugs mingqi, zhujieshen, zhuzishen grown in Sichuan]. PMID- 2964315 TI - [Effects of neonatal administration of monosodium glutamate on acupuncture analgesia and beta-endorphin-immunoreactive neurons in the rat hypothalamic arcuate nucleus]. PMID- 2964316 TI - Atrial natriuretic peptide: blood levels in human disease and their measurement. AB - The atrial hormonal system consists of 126 amino acid-containing prohormone (proANP) stored in the secretory granules of atrial myocytes and 28 amino acid containing hormone (ANP) that is secreted into the bloodstream in response to raised atrial pressure. ANP participates in the homeostasis of body fluid volume through its main receptor-mediated effects; natriuresis, inhibition of renin and aldosterone secretion, and vasodilation. It counteracts the renin-angiotensin system with the putative primary role of regulating the circulating blood volume. Although in man, the physiologic volume stimuli lead to relatively modest increases of ANP secretion, its plasma level undergoes striking changes in pathology. Marked elevations in conditions accompanied by fluid retention, most conspicuously in heart failure and renal failure, have been explained as a compensatory reaction to volume overload. The recent data suggest a decreased target organ responsiveness as one of the causes of a relative inefficiency of the high circulating levels of ANP in inducing an appropriate natriuresis in these volume overload conditions. The well established radioimmunoassay and the more recent methods of plasma ANP measurement are reviewed, and the authors' results with a commercial RIA are presented. PMID- 2964317 TI - Dissection of the left main coronary artery: a complication of PTCA to the anterior descending artery. AB - This case report describes a patient with severe discrete left anterior descending coronary artery atheroma who underwent percutaneous transluminal coronary angioplasty. During the procedure, dissection occurred in a superior atheromatous plaque of the left main coronary artery and needed immediate surgery. Special care should be taken in the selection and angioplasty of patients with concomitant left main artery disease. PMID- 2964318 TI - Sebum production is increased in Behcet's syndrome and even more so in rheumatoid arthritis. AB - Sebum production is under hormonal control. We had shown that male sex is associated with more severe disease in Behcet's syndrome and the acneiform skin lesion of this disorder is not different from ordinary acne, an androgen dependent lesion. Sebum excretion rate was higher in patients with Behcet's syndrome than in healthy controls, children and patients with ankylosing spondylitis. On the other hand, patients with rheumatoid arthritis had high levels of sebum excretion rate comparable to those found in patients with acne vulgaris. These suggest the presence of a sebotrophic hormone and/or other hormonal effects in Behcet's syndrome and rheumatoid arthritis. PMID- 2964319 TI - Atrial natriuretic factor (ANF) in experimental and human hypertension. AB - The plasma levels of immunoreactive (IR)-ANF have been evaluated by radioimmunoassay in several models of experimental hypertension and in human hypertension. In all models of experimental hypertension so far studied, the plasma levels of IR-ANF are consistently increased. This is accompanied by a decrease, at certain time intervals, of the IR-ANF levels in the left atrium. In human essential hypertension, the plasma levels of IR-ANF are not increased except in the severe form (diastolic blood pressure above 110 mmHg). In renovascular hypertension, the peripheral levels of IR-ANF are not different from the normal levels but are increased above normal in aortic blood. PMID- 2964320 TI - Relevance of echocardiographic left ventricular hypertrophy in mild hypertension. AB - The current diagnostic criteria for uncomplicated mild hypertension inadequately distinguish the subgroup actually at increased risk for cardiovascular sequelae, from the large group not at increased risk, and therefore unlikely to benefit from current pharmacological intervention. Although finding of electrocardiographic left ventricular hypertrophy indicates increased risk, the ECG lacks sufficient sensitivity to be diagnostically useful in mild hypertension. It can be argued that echocardiographic screening for LV hypertrophy may better detect the subpopulation of mildly hypertensive patients at increased risk. The known relationships between ambulatory BP, LV hypertrophy and prognosis support this hypothesis. Further study is required, however, before echocardiography can be advocated for routine application to the diagnosis and management of mild hypertension. PMID- 2964321 TI - Diabetes, the way ahead. PMID- 2964322 TI - The dawn phenomenon: nocturnal blood glucose homeostasis in insulin-dependent diabetes mellitus. PMID- 2964323 TI - De natrio diabeticorum. Increased exchangeable sodium in diabetes. PMID- 2964324 TI - Carbohydrate and lipid metabolism of skeletal muscle in type 2 diabetic patients. AB - Peripheral hyperinsulinaemia is the cause of metabolic changes that might contribute to the high incidence of macrovascular disease in patients with diabetes mellitus. In order to test this hypothesis muscle biopsies from 12 Type 2 diabetic patients and 14 age and sex matched non-diabetic patients, undergoing minor surgery, were obtained. The diabetic patients had significantly elevated fasting serum insulin (0.29 +/- 0.05 vs 0.06 +/- 0.03 nmol-1) and glucose (8.3 +/ 1.5 vs 4.6 +/- 0.5 mmol-1) and HbA1 levels (8.4 +/- 0.4 vs 5.0 +/- 0.2 per cent). The fasting and 2-h postprandial C-peptide levels were 0.99 +/- 0.25 vs 0.39 +/- 0.12 and 3.12 +/- 0.75 vs 1.09 +/- 0.34 nmol/l, respectively. The diabetic patients showed a marked elevation of triglyceride in the striated muscle biopsies compared to the non-diabetic controls (290 +/- 52 vs 48 +/- 6 mumol/g wet weight, p less than 0.001). Moreover, the activities of glucose-6 phosphate dehydrogenase (0.25 +/- 0.03 vs 0.13 +/- 0.01 U/g wet weight) and malic enzyme (0.15 +/- 0.01 vs 0.05 +/- 0.01 U/g wet weight), necessary for lipid synthesis, were significantly increased (both p less than 0.001) in the diabetic patients while the glycolytic enzymes, hexokinase (0.65 +/- 0.09 vs 1.82 +/- 0.11 U/g wet weight), pyruvate kinase (7.3 +/- 0.9 vs 13.2 +/- 0.9 U/g wet weight), phosphofructokinase (1.3 +/- 0.2 vs 2.6 +/- 0.2 U/g wet weight), and alpha glycerophosphate dehydrogenase (7.3 +/- 0.5 vs 12.5 +/- 0.7 U/g wet weight) were decreased (all p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964325 TI - The importance of the time interval between insulin injection and breakfast in determining postprandial glycaemic control--a comparison between human and porcine insulin. AB - This study assessed the effect of the time interval between insulin injection and breakfast in determining subsequent postprandial glycaemic control and also whether this differed between highly purified porcine insulin and human insulin (crb) in six diabetic patients (age range 24-36 years, duration of diabetes greater than 10 years) usually treated with twice daily Actrapid MC and Monotard MC and with stable insulin requirements and diabetic control. On separate mornings each patient was given, after an overnight fast, their usual dose of either Actrapid MC and Monotard MC or Humulin S and Humulin Zn injected 5, 20, or 40 min before a standard breakfast. The postprandial glycaemic profile was not significantly different at any of the three time intervals with Actrapid MC and Monotard MC. However, with the human insulin the profile was significantly better at the 40 min interval than at the 5 min interval (p less than 0.05) and this was also better than any of the profiles with the porcine insulin, there being a significant difference between the two types of insulin (p less than 0.05). These findings suggest that the time interval between insulin injection and breakfast may be more important with human insulin than with porcine insulin. PMID- 2964326 TI - Natural history of pancreatic islet B-cell function in type 2 diabetes mellitus studied over six years by homeostasis model assessment. AB - Islet B-cell function and insulin sensitivity were estimated with the aid of a mathematical model from repeated fasting plasma glucose and insulin measurements over a 6 year period in 131 patients with type 2 diabetes mellitus who could be managed satisfactorily on dietary therapy alone. They presented between the ages of 40 and 69 years, and were studied before and after one year of treatment, and then at regular intervals from 12 until 72 months later. A method of averaging the individual trends by means of a linear model regression technique was used to assess the progression of their diabetes. Dietary management over the first 12 months resulted in weight loss from 118% to 106% average body weight, and improved insulin sensitivity from 26% to 40% of normal (p less than 0.001). From 12 to 72 months, the fasting plasma glucose rose at a mean rate of 0.23 mmol/l per year (p less than 0.001) despite a rate of weight loss of 0.2% average body weight per year (p less than 0.01). The estimated islet B-cell function, expressed as a percentage of normal, decreased significantly (p less than 0.05) at a rate of 1.5% per year, with no statistically significant change in insulin sensitivity. Extrapolation suggests the reduction in B-cell function predated the departure of fasting plasma glucose from the normal range. PMID- 2964327 TI - Alpha glucosidase inhibition in the treatment of non-insulin-dependent diabetes mellitus. AB - Two studies of the new alpha-glucosidase inhibitor, miglitol, in patients with non-insulin-dependent diabetes mellitus (NIDDM) are reported. In the first, 13 patients, poorly controlled on sulphonylureas, received miglitol 50mg three times daily for 4 weeks. Post-prandial blood glucose was reduced after breakfast, lunch, and tea compared with placebo (p less than 0.05-0.01) but there was no improvement in fasting blood glucose, serum fructosamine or haemoglobin A1. In a dose-response study the effect of a single dose of miglitol (0,50,100,150 or 200mg) on post-prandial glycaemia after a test breakfast was assessed in 20 patients with mean +/- SEM fasting blood glucose 9.9 +/- 0.4 mmol/l. With 50mg miglitol, there was a significant reduction in blood glucose from 30 to 120 min post-prandially compared with placebo. Increasing doses of miglitol further depressed the post-prandial rise in blood glucose and with 200mg there was no significant change from fasting levels. Side-effects were limited to flatus and loose stools particularly with the higher doses but were not severe. Miglitol effectively reduces post-prandial blood glucose rise in NIDDM with as little as 50mg but there is considerable individual variation. Larger doses may be necessary in patients already poorly controlled on sulphonylureas. PMID- 2964328 TI - Hormonal and metabolic changes during hypothermic coronary artery bypass surgery in diabetic and non-diabetic subjects. AB - Hormonal and metabolic responses to hypothermic coronary artery bypass grafting (CABG) were studied in three groups: 8 non-diabetic patients, 8 patients with non insulin-dependent diabetes mellitus (NIDDM) given a glucose pump priming solution and 8 NIDDM patients given a non-glucose infusion. There were no significant differences in stress hormone responses between NIDDM and non-diabetic patients, with adrenaline concentrations rising 10-fold, noradrenaline 4-fold and cortisol 2 to 3-fold. Glucagon rose significantly during bypass only in the NIDDM patients who did not receive a glucose prime. Comparable marked hyperglycaemia was seen in both glucose primed groups during bypass and exclusion of glucose from the prime in NIDDM patients prevented this major rise. Postoperatively, the rise in insulin in the glucose primed NIDDM patients contrasted with the slower rise in the non glucose primed NIDDM patients who were also hyperglycaemic by this stage. Perioperative hyperglycaemia in NIDDM patients undergoing CABG can be prevented by using a non-glucose priming solution and by giving insulin infusion, particularly postoperatively. PMID- 2964329 TI - Microvascular disease and limited joint mobility in diabetes. A comparison of fibrinolysis and prostacyclin in diabetes and systemic sclerosis. AB - The pathogenesis of Limited Joint Mobility (LJM) in diabetes is unknown, but the abnormality is said to be associated with an increased incidence of microangiopathy. To examine the possibility that LJM may be a manifestation of microvascular disease, fibrinolysis and blood prostacyclin metabolites were measured in diabetic patients with and without LJM, and compared with patients who have systemic sclerosis (PSS). The concentrations of plasma fibrinogen (p less than 0.01) and Plasminogen Activator Activity (PAA) (p less than 0.02) were increased in both diabetic groups, compared with non-diabetic controls, and were of similar magnitude to the changes observed in PSS. Antithrombin III (AT III) activity was significantly lower in diabetic patients with LJM than in those without LJM, but not significantly different from controls. Prostacyclin concentrations were raised in PSS patients compared to controls (p less than 0.02) but were not raised in diabetic subjects. Thus abnormal fibrinolysis was demonstrated in diabetic patients compared to non-diabetic controls, and prostacyclin concentrations in diabetics were lower than in PSS patients. The diabetic patients with LJM had lower AT III activity than diabetics without LJM, but overall a convincing difference between the two groups of diabetics was not proven. No sure inferences could be drawn on the vascular aetiology of LJM, while the impaired prostacyclin activity might contribute to the development of diabetic microangiopathy. PMID- 2964330 TI - Development, validation and application of computer-linked knowledge questionnaires in diabetes education. AB - Multiple choice questionnaires (MCQs) capable of being marked manually or by a newly developed optical mark reader, or by use of an inexpensive inter-active microcomputer system have been developed for the separate assessment of insulin dependent and non-insulin-dependent patient knowledge. Forty-six insulin-related and non-insulin-related multiple choice questions covering six main areas of knowledge were constructed for inclusion into draft questionnaires. From the responses of a total of 180 completed questionnaires, piloted in 18 randomly selected clinics in 14 Regional Health Authorities in England, psychometric analysis was performed to determine reliability, discrimination coefficients, and facility indices. Seventy-three per cent of insulin-dependent diabetic patients (IDDM) and 92% of non-insulin-dependent diabetic patients (NIDDM) MCQ correct options had facility indices within the acceptable range of 30 to 90%. 82% IDDM and 93% NIDDM correct options had discrimination coefficients exceeding 0.2. Questionnaire reliability (internal consistency) using the Kudor-Richardson (KR20) formula was IDDM 0.87 and NIDDM 0.82. Evidence in support of the IDDM questionnaire's criterion validity was based on significant differences (p less than 0.05) identified between a number of knowledge area scores stratified according to HbA1 levels. Prescriptive correction for screen display and automatic hard copy feedback was designed for both incorrect and omitted question options, providing both educational (patient) and analytical (clinic) documentation. Both technical and psychometric properties of these knowledge assessment instruments should be acceptable for diabetic knowledge evaluation and instruction. PMID- 2964331 TI - Diuretics and hyperkalaemia in diabetic ketoacidosis. AB - Diabetic ketoacidosis (DKA) often presents with hyperkalaemia. We investigated whether it was more likely in patients taking potassium-retaining diuretics. A retrospective survey of all patients (552 cases) presenting in DKA between 1974 and 1984 was undertaken. Initial biochemical data were compared for patients recorded as taking potassium-retaining diuretics (7 cases) at the time of presentation with those taking potassium-losing diuretics (13 cases), and age matched control groups were selected from those who presented in DKA but were not taking diuretics. There was no significant difference in initial serum potassium levels between the diuretic treated groups. The serum sodium was higher in the control group than in the potassium losing group (p = 0.045) and the serum urea significantly lower (p = 0.045). We conclude that potassium-retaining diuretics do not predispose to hyperkalaemia in diabetic ketoacidosis. PMID- 2964332 TI - The feasibility of a potentially 'ideal' system of integrated diabetes care and education based on a day centre. AB - A new system of clinical and educational care designed to replace traditional diabetic clinics is described. The overall aims were to provide adequate consultation time, patient access according to diabetic and social needs, minimal waiting time, continuity with experienced staff and objective based learning programmes in an environment suited to learning. This was achieved by changing the role of the diabetes nurse specialist from undertaking delegated tasks to providing primary consultative care, reorganization of existing staff and provision of a purpose designed unit. This gave sufficient flexibility to arrange daily early morning and weekly evening clinic sessions for routine diabetes counselling and medical audit with primary education scheduled at other times. A year's experience showed that our major targets had been met, with the provision of adequate consultation time, halving of waiting time, ease of patient access and continuity with either the doctor or nurse in consulting role. Default rates have fallen and patient and staff morale has improved substantially. Apart from an underestimate of receptionist and a small increase in technician hours, these changes have been achieved within the predicted small revenue cost. PMID- 2964333 TI - A comparison of arterial and non-arterialized capillary blood gases in diabetic ketoacidosis. AB - The results of acid-base and blood gas estimations in arterial and non arterialized capillary blood have been compared in samples obtained simultaneously from patients presenting with diabetic ketoacidosis. Highly significant correlations were obtained for pH, pCO2 and bicarbonate measurements. Small but significant differences were observed with capillary pH slightly lower and capillary pCO2 and bicarbonate slightly higher than arterial values. These differences were of no clinical significance. Non-arterialized capillary samples are a reliable indicator of acid-base status in this form of metabolic acidosis and are preferable to repeated arterial puncture. PMID- 2964334 TI - Renal failure in diabetics in the UK: deficient provision of care in 1985. Joint Working Party on Diabetic Renal Failure of the British Diabetic Association, the Renal Association, and the Research Unit of the Royal College of Physicians. AB - Two surveys of diabetic renal failure were conducted in 1985. The first survey was designed to identify all diabetic patients having advanced renal failure (serum creatinine greater than 500 mumol/l and/or blood urea greater than 25 mmol/l in six selected health regions of the UK. The study identified 181 such patients indicating that approximately 10 people in every million of the population were affected. It is calculated that there were about 580 patients having diabetic renal failure in the whole of the UK. The majority of the patients (78%) were considered by their physicians to be suitable for renal support treatment. Those who were excluded were considered unsuitable chiefly on account of old age or cardiac problems. In the second survey all 66 renal units in the UK reported the number of diabetic patients starting renal support treatment in 1985. There were 273 patients, suggesting that there may be a shortfall when compared with the number of diabetics likely to need treatment. The Working Party recommends that diabetics who develop renal failure should always be considered for treatment by dialysis or transplantation, and that appropriate resources should be made available for them. PMID- 2964335 TI - Hypertriglyceridaemia and diabetes mellitus: cause or effect? AB - Two patients with hypertriglyceridaemia which was diagnosed several years before the onset of diabetes are described. In the first case a 54-year-old man presented with hypertriglyceridaemia and normal glucose tolerance. After 4 years he developed severe hypertriglyceridaemia (22 mmol/l) which was first ascribed to poor compliance but soon afterwards diabetes was diagnosed and good blood glucose control minimized the hypertriglyceridaemia. The second patient, a female, aged 27 years, with the polycystic ovarian syndrome, presented with eruptive xanthoma after several years of mild hyperlipidaemia. There was severe hyperlipidaemia (triglyceride 140 mmol/l, cholesterol 42 mmol/l) which required urgent plasmapheresis. Diabetes was diagnosed and treated with insulin at this time but the patient was taking the oral contraceptive pill and also had a deficiency of apolipoprotein CII. PMID- 2964336 TI - Diabetic ketoacidosis during NovoPen therapy. AB - To improve flexibility and acceptability of diabetic management five patients (age 15-26 years) were changed to NovoPen therapy (Human Actrapid Insulin 3 x daily + Human Ultratard Insulin at night). Diabetic ketoacidosis developed within 2-4 months of commencing this regimen and in one patient twice in only 4 weeks. All failed to increase their insulin dose in response to hyperglycaemia and two patients omitted insulin when they did not take meals believing this to be appropriate irrespective of the prevailing blood glucose. The NovoPen regimen may increase the risk of diabetic ketoacidosis in patients who do not monitor and control their diabetes assiduously and should not be seen as a solution for poorly motivated or ill-educated patients. PMID- 2964337 TI - British Diabetic Association holidays--what are they worth? AB - A postal survey among families of 63 children attending two BDA educational holidays studied their expectations before (Questionnaire 1) and experiences 3 months afterwards (Questionnaire 2). Of 47 families replying to both questionnaires (22M, 25F; mean age 8.4 years, range 5-13.8 years), expectations expressed by parents for their children were that they gain in confidence and independence (n = 36), mix with other diabetic children (n = 32), learn to self inject (n = 27), improve diet (n = 23), and have a good holiday (n = 10). Thirty five families subsequently reported that their expectations had been met. After the holiday, although there was no change in frequency of blood or urine testing, there was an increase in children doing tests on their own (Q1: 15, Q2: 28, p = 0.003); more parents reported confidence in their child recognizing a hypoglycaemic episode (Q1: 29, Q2: 39, p = 0.001); and there was a sustained increase in independence with insulin injections (Q1: 29, Q2: 35, p = 0.042). This study provides evidence that BDA holidays benefit parents and their children, and do so mainly by increasing the children's confidence and independence in diabetic care. PMID- 2964339 TI - What's in a name? Microalbuminuria/albuminuria. PMID- 2964338 TI - Spreading the message further. PMID- 2964340 TI - Treatment of ordinary and penicillinase-producing strains of Neisseria gonorrhoeae in Mexico City. AB - Since being recognized in 1976 and 1983, respectively, penicillinase-producing strains of Neisseria gonorrhoeae (PPNG) and chromosomally mediated resistance (CMRNG) have attained a worldwide distribution. The high endemicity of both types of resistance in some regions precludes the continued routine use of procaine penicillin (APPG) as treatment for gonorrhea. In this study, we have evaluated 72/216 men with uncomplicated gonococcal urethritis at the venereal clinic in Mexico City. These men were part of a blinded randomized comparative study for treatment with group (P) penicillin having 33 patients and group (S) spectinomycin having 39. Efficacy with (P) was 24/33 (72.7%); nine failures retreated and were cured with spectinomycin. Efficacy with (S) was 35/39 (89.7%); four failures retreated and were cured with cefotaxime. We found correlation between MICs and resistance; all the strains with MICs of greater than or equal to 1.0 mcg/ml of penicillin failed to be cured, the MICs of greater than or equal to 32 mcg/ml of spectinomycin failed to be cured. The overall resistance to both regimens was 23/72 to penicillin (31.9%) (22 PPNG and one CMRNG) and 4/72 (5.5%) to spectinomycin. PMID- 2964341 TI - Is the glomerular capillary hydraulic pressure increased in moderately hyperglycemic rats on normal protein intake? PMID- 2964342 TI - Effects of immunostimulation on host survival in A/J mice with transplantable C1300 neuroblastoma. PMID- 2964343 TI - Chronic fixed drug eruption caused by acetaminophen. AB - A case of chronic fixed drug eruption resembling parapsoriasis en plaques is reported, which presented with persistent, stable lesions that were present for seven months before the diagnosis was established. The patient's skin cleared totally with avoidance of acetaminophen and flared prominently with re-exposure. Distribution of the recurrence was different: some previous sites had apparently become refractory and remained clear, some involvement had recurred in the same site, and new areas of involvement had appeared, causing the eruption to "wander," as is often seen in acute fixed drug eruption due to acetaminophen. PMID- 2964344 TI - Erythromycin 2 percent gel in the treatment of acne vulgaris. AB - A gel formulation of erythromycin 2 percent was compared with its vehicle in a double-blind multicenter study involving patients with mild to moderate acne vulgaris. In an analysis of 187 patients treated twice daily for 8 weeks, erythromycin 2 percent gel proved to be significantly more effective than vehicle in reducing the numbers of both inflammatory and noninflammatory lesions. After 8 weeks, 60 percent of erythromycin-treated patients had good or excellent responses compared with 36 percent of those using vehicle (p = 0.001); the lesions in two patients using erythromycin were completely cleared. The majority of patients had a favorable impression of the cosmetic characteristics of the gel formulation. PMID- 2964345 TI - [Investigation on the reduction of intussusception under B-mode ultrasonographic monitoring]. PMID- 2964346 TI - [Radiological diagnosis of parathyroid adenoma (a review of 50 cases)]. PMID- 2964347 TI - [X-ray analysis of the chest films in 9 cases of primary penicilliosis]. PMID- 2964348 TI - [The computed tomographic anatomy of the nasopharyngeal region]. PMID- 2964349 TI - [CT of the nasopharynx]. PMID- 2964350 TI - [Computed tomographic metrizamide myelography in the diagnosis of syringohydromyelia (report of 26 cases)]. PMID- 2964351 TI - [CT diagnosis of infratentorial angioblastomas]. PMID- 2964352 TI - [Transcatheter plugging of patent ductus arteriosus]. PMID- 2964353 TI - [A new method percutaneous lung needle biopsy with tissue adhesive for the prevention of complications]. PMID- 2964354 TI - [Giant lymph node hyperplasia]. PMID- 2964355 TI - [Transiliac lengthening: a radiologic study]. PMID- 2964356 TI - [Occupationally-induced dyshidrosiform dermatitis of the hands following contact with cacti. Case report]. AB - A patient, employed in a plant nursery where cacti are grown, developed wide spread dyshidrotic dermatitis of the hands. This patient was histologically studied and cactus spines were found in several biopsied vesicles. PMID- 2964357 TI - [Contact allergy to dexpanthenol]. AB - A case report is given of a 56-year-old female patient with a postthrombotic leg ulcer and stasis dermatitis, in whom contact dermatitis to a dexpanthenol containing ointment was first described in 1965. Recent allergy testing confirmed this finding and revealed positive reactions to the pure substance dexpanthenol as well as to two additives. Based on this case history the importance of dexpanthenol as an allergen is discussed. PMID- 2964358 TI - [Contact allergy to dicyclohexylcarbodiimide and N-hydroxyphthalimide]. AB - We report a case of a female laboratory worker in the chemical industry who acquired contact allergy to dicyclohexylcarbodiimide and N-hydroxyphthalimide. Both substances are used in the chemical synthesis of peptides. So far there have only been a few reports concerning contact allergy to these substances. PMID- 2964359 TI - A simple technique for recording the electromyogram of the external abdominal oblique muscle in the newborn. AB - A technique for recording activity in the external abdominal oblique muscles of the human newborn is described. It uses simple, self-adhesive, surface electrodes arranged to reduce electrocardiographic interference. This method is applicable to studies of the respiratory patterns of very pre-term babies undergoing intensive medical care. PMID- 2964360 TI - Desulfation of 3,5,3'-triiodothyronine sulfate by microsomes from human and rat tissues. AB - Subcellular preparations from rat liver, brain, and kidney and from human liver were tested for their ability to desulfate T3 sulfate (T3SO4). Activity was found associated with the microsomal fraction: rat liver was the most active, hydrolyzing 76 pmol/min.mg protein of T3SO4 while preparations from rat kidney and brain were about 1/5 and 1/20 as active. Microsomal preparations from human liver obtained at autopsy were as active as fresh rat preparations. Thyroxine sulfate was not an active substrate. Microsomes prepared with dithiothreitol and EDTA in order to detect deiodinating activity maintained T3SO4-desulfating activity. Cytosolic preparations containing arylsulfatase activities failed to desulfate T3SO4. Estrone sulfate, dehydroepiandrosterone sulfate, and nitrophenyl sulfate are known substrates for microsome-associated arylsulfatase activities, and these compounds were found to inhibit hydrolysis of T3SO4 to various extents. Of these competing sulfatase substrates, only dehydroepiandrosterone sulfate inhibits T3SO4 desulfation completely. In order to determine whether desulfation occurs in intact cells, isolated hepatocytes were incubated in the presence of 7 and 54 microM T3SO4. These cells were found to hydrolyze 1-1.5% of the sulfate ester/h for up to 3 h. The demonstration of this activity raises the possibility that these hepatic cells may be able to reactivate T3SO4, which has generally been regarded as an irreversibly inactivated metabolite. PMID- 2964361 TI - Inability of a mouse cell line transformed to produce biologically active recombinant human insulin-like growth factor I (IGF-I) to respond to exogenously added IGF-I. AB - A plasmid expression vector encoding human insulin-like growth factor I (hIGF-I) in the form of a 97-amino acid precursor protein containing the first 27 amino acids of prebovine GH and the 70 amino acids of hIGF-I has been used to transform mouse L cells. A stably transformed mouse L cell clone has been isolated which expresses and secretes hIGF-I. The secreted peptide comprises 3% of the protein in conditioned medium. IGF-I can be purified to homogeneity in 2 chromatographic steps. One liter of conditioned medium yields approximately 200 micrograms purified peptide. Amino-terminal sequence analysis confirms that the signal peptide has been proteolytically hydrolyzed from the precursor protein before secretion to form [Ala0]hIGF-I. The recombinant peptide and serum-derived hIGF-I are equipotent as inhibitors of the binding of [125I]IGF-I to the type 1 receptor of human placenta and to a crude preparation of acid-stable human serum binding proteins. The peptides are equipotent in 2 in vitro assays, the stimulation of the rate of 2-[1,2-N-3H]deoxyglucose transport in BC3H1 cells and the stimulation of [methyl-3-3H]thymidine incorporation into DNA in A10 cells. In contrast to a control mouse L cell line, DNA synthesis in the [Ala0]IGF-I-secreting line is completely unresponsive to [Thr59]IGF-I, while it responds normally to calf serum (10%). Thus, the [Ala0]IGF-I-secreting line is selectively desensitized to IGF-I. The binding of [125I]IGF-I to both lines is identical, indicating that the loss of responsiveness to IGF-I is not due to a loss of cell surface receptor. The ability to render mouse L cells unresponsive to IGF-I is transferred in the conditioned medium of the [Ala0]IGF-I-secreting cell line. In addition, pretreatment of control cells with [Thr59]IGF-I (10 nM) results in attenuation of the response to a subsequent dose of IGF-I. These data indicate that prolonged exposure to high levels of IGF-I may cause a postreceptor-mediated desensitization to IGF-I. Alternatively, IGF-I may promote secretion of an inhibitor of IGF-mediated DNA synthesis. PMID- 2964362 TI - Properties of [3H]1-desamino-8-D-arginine vasopressin as a radioligand for vasopressin V2-receptors in rat kidney. AB - [3H]1-Desamino-8-D-arginine vasopressin [3H] DDAVP was assessed as a radioligand for vasopressin V2-receptors by studying its membrane-binding characteristics and in vitro autoradiographic localization in rat kidney, a rich source of V2 receptors. [3H]DDAVP bound specifically to a single class of high affinity, low capacity sites in rat medullopapillary membranes. Specific [3H]DDAVP binding at 25 C reached equilibrium after 2 h of incubation and was saturable and linear with protein concentration up to 2.2 mg/ml protein. Saturation analysis gave an equilibrium dissociation constant (Kd) of 0.76 nM. Displacement of [3H]DDAVP binding by unlabeled arginine vasopressin (AVP) and related analogs gave the following order of potency, consistent with that expected for a V2-receptor: DDAVP approximately equal to AVP approximately equal to 1-desamino-AVP greater than lysine vasopressin greater than oxytocin greater than [1-(beta-mercapto beta, beta-cyclopentamethylene-propionic acid, 2-(O-methyl)tyrosine]AVP. The C terminal metabolites of AVP, (pGlu4Cyt6)AVP-(4-9), and (pGlu4Cyt6)AVP-(4-8) did not displace [3H]DDAVP binding. No degradation of [3H] DDAVP during incubation could be detected by HPLC analysis. In vitro autoradiography of [3H]DDAVP binding to rat kidney sections showed a very dense localization of displaceable binding over inner and outer medulla, with a much lower density in cortex, consistent with the known major localization of V2-receptors on renal collecting tubules. These studies suggest that [3H]DDAVP is a suitable radioligand for labeling V2 receptors and may be useful in the characterization of vasopressin receptor subtypes in a variety of tissues and in purification of the V2-receptor. PMID- 2964363 TI - In vivo regulation of granulosa cell somatomedin-C/insulin-like growth factor I receptors. AB - The characteristics and regulation of the murine granulosa cell type I insulin like growth factor (IGF) receptor under in vivo conditions were studied. In vivo treatment of immature hypophysectomized diethylstilbestrol-treated rats with increasing doses (0.3-30 microgram/rat, twice daily) of FSH for 72 h resulted in dose-dependent increments in specific granulosa cell somatomedin-C (Sm-C)/IGF-I binding, peaking (5150 +/- 350 cpm/3 x 10(5) cells) at the 10 micrograms/rat (twice daily) dose level to yield a 2.6-fold increase relative to that in untreated controls. This FSH (10 micrograms/rat, twice daily) effect proved time dependent; the first significant (P less than 0.05) increase in binding (3670 +/- 150 cpm/3 x 10(5) cells) was noted after 48 h of treatment (1.6-fold increase). Significantly, little or no variation was observed for basal Sm-C/IGF-I binding over the course of the experiment, suggesting that this component of Sm-C/IGF-I receptor complement may be independent of the trophic influence(s) of the pituitary gland. Equilibrium competition studies carried out with granulosa cells derived from both control and FSH-treated rats revealed linear Scatchard plots consistent with a single class of noninteracting binding sites, a 2.8-fold increase in FSH-associated Sm-C/IGF-I-binding capacity, but not affinity (Kd control, 1.9 +/- 0.3 nM; kd FSH, 2.6 +/- 0.9 nM). Limited specificity studies of the FSH-induced receptor revealed related peptides to compete for Sm-C/IGF-I binding with a relative rank order of potency of Sm-C/IGF-I much greater than multiplication-stimulating activity greater than insulin, a pattern compatible with a type I IGF receptor. A series of other polypeptides, including porcine relaxin, porcine proinsulin, epidermal growth factor, basic fibroblast growth factor as well as transforming growth factor-alpha and -beta (TGF beta) were nonreactive. Significantly, the induced type I IGF receptor proved functionally coupled to granulosa cell proteoglycan biosynthesis. The ability of FSH (10 micrograms/rat, twice daily) to enhance granulosa cell Sm-C/IGF-I binding was significantly (P less than 0.05) up-regulated (1.53-fold amplification) by ovine GH (100 micrograms/rat, twice daily); a down-regulatory effect (64% inhibition) was observed for a potent GnRH agonist [( D-Ala6,Des-Gly10]GnRH ethyl amide; 25 micrograms/rat, twice daily). Once induced, the Sm-C/IGF-I receptor of the granulosa cell required the continued presence of either FSH or LH for its maintenance; the lactogenic receptor agonist PRL had no effect.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2964364 TI - Atrial natriuretic peptide inhibits the stimulation of aldosterone secretion but not the transient increase in intracellular free calcium concentration induced by angiotensin II addition. AB - The effect of atrial natriuretic peptide (ANP) on cellular calcium metabolism was evaluated in bovine adrenal glomerulosa cells stimulated by agonists that use the Ca2+-phosphoinositide messenger system. The calcium-sensitive probe aequorin was used to measure intracellular free calcium concentration, and the aldosterone secretory rate was simultaneously monitored. ANP did not block the calcium transient induced by beta-[Asp1]angiotensin II (beta-[Asp1]AII), an AII analog, but markedly reduced the stimulated rate of aldosterone secretion. Consistent with these findings, radiolabeled 45Ca efflux stimulated by AII and carbachol was not altered by the concurrent addition of ANP. These results indicate that ANP has no effect on the phosphoinositide-mediated calcium transient and the associated rise in cellular calcium efflux, suggesting that these parameters of calcium metabolism are not the locus of ANP's inhibitory action. PMID- 2964365 TI - Atrial natriuretic factor inhibits luteinizing hormone secretion in the rat: evidence for a hypothalamic site of action. AB - The presence of atrial natriuretic factor (ANF) immunoreactivity and receptors for ANF in the median eminence, hypothalamus, and anterior pituitary gland suggests a role for the peptide in the hypothalamic control of anterior pituitary function. In conscious ovariectomized female rats, transient elevation of plasma levels of ANF by volume loading, a stimulus known to release endogenous ANF from the heart, or by bolus iv injection of 0.1, 1.0, or 10 micrograms synthetic ANF failed to result in altered circulating levels of LH or GH. Constant iv infusion of ANF for 30 min, such that 2- to 3-fold elevations in plasma ANF were detected by RIA resulted, however, in significant inhibition of LH release in ovariectomized female rats (0.05 and 0.1 micrograms ANF/kg.min) and orchidectomized male rats (0.1 microgram ANF/kg.min). It was unlikely that this effect was exerted at the level of the anterior pituitary, since ANF failed to alter basal or LHRH-stimulated LH release from cultured anterior pituitary cells in vitro and since iv infusion of 0.1 microgram ANF/kg.min failed to alter pituitary responsiveness in vivo to a 10-ng bolus injection of LHRH. Significant inhibition of LH secretion was also observed after third cerebroventricular injection of 1.0 or 2.0 nmol ANF. As with iv infusion, central administration of ANF failed to significantly alter GH secretion. LHRH release from median eminence explants incubated in vitro in the presence of dopamine (60 or 120 microM) was inhibited by 10(-7) M ANF, suggesting a median eminence site of action of the peptide. Finally, an opiate involvement in the mechanism of ANF's action was suggested, since naloxone (0.5 mg, iv, followed by a 60-min infusion of an additional 1 mg) completely blocked the ability of ANF (0.1 or 0.5 microgram/kg.min, infused over the last 30 min of naloxone administration) to inhibit LH release. These data suggest that ANF can act centrally to alter the hypothalamic control of gonadotropin secretion, possibly by interacting with central dopaminergic and peptidergic systems. They further suggest actions of ANF within the brain unrelated to its previously described effects on fluid and electrolyte homeostasis. PMID- 2964366 TI - Mutations affecting transport and stability of lysosomal enzymes. AB - The biosynthesis, post-translational processing and receptor-mediated transport of lysosomal enzymes will be briefly summarized. Mutations affecting the transport or the stability of a lysosomal enzyme but not its catalytic properties can result in a lysosomal storage disorder. Mutations causing a loss of catalytic activity may in addition affect transport or stability. Such mutations should not be classified as transport or stability mutations. Prototypes for transport and stability mutations are I-cell disease and late onset forms of metachromatic leukodystrophy. PMID- 2964367 TI - Interferon removes its own receptors as it blocks the division of Daudi cells. AB - The Burkitt-derived line, Daudi, whose proliferation is inhibited by human alpha interferon (IFN-alpha), was treated with 125I-labelled recombinant human IFN alpha A. After separation from unbound ligand, cell-bound IFN was extracted with the detergent digitonin yielding soluble and insoluble complexes of IFN and receptor, together with a certain amount of uncomplexed IFN. 1. Soluble complexes were stable enough to be separated from uncomplexed IFN by permeation chromatography. Treatment of soluble complexes with the bifunctional reagent, disuccinimidyl suberate, yielded a radioactive product separating with an Mr of 130,000 on electrophoresis in sodium dodecyl sulphate. Similar complexes could be recovered with sodium dodecyl sulphate from the digitonin-insoluble residue, treated with the bi-functional reagent. 2. The total (soluble and insoluble) of complexed IFN obtained after digitonin extraction was a constant fraction (0.62) of the total cell-bound radioactivity, being independent of the concentration of IFN added to the cells (less than pM to greater than nM), and of the time of incubation (1 min to 20 h). However, between 30 min and 3 h of incubation, the insoluble complex increased, at the expense of the soluble complex, and there appeared a cellular pool of degraded ligand. From 3 h to 20 h the distribution of ligand-derived radioactivity remained constant while the total amount decreased to less than 10% of its value at 30 min. This decrease in binding was matched by the appearance of an equivalent quantity of radiolabelled fragments in the culture medium. 3. The inhibition of cellular division due to IFN was shown to be coincident with the disappearance of cellular binding and with the cell-mediated degradation of receptor-complexed IFN. We propose that IFN removes its own receptor and, in doing so, blocks a linked function necessary for the stimulated growth of Daudi cells. PMID- 2964368 TI - Electrostatic interactions in the cellular dynamics of the interferon-receptor complex. AB - Using membrane preparations of the interferon receptor, prepared from cells of the Burkitt line, Daudi, we have examined the binding of three human recombinant alpha-interferons. 1. We discovered a binding titration of the interferons IFN alpha A and IFN-alpha D in the pH range 6-9. Receptor binding, negligible at pH 6, rises to a maximum close to pH 9. We have shown that binding of IFN-alpha A at basic pH is to the same receptors as at neutrality and that IFN-receptor complexes extracted with digitonin are more stable at basic pH than they are at neutrality. 2. The recombinant interferon, IFN-alpha B, shows little change of binding in the pH range 6-9. At its basic optimum the binding of IFN-alpha A approaches that of IFN-alpha B, while at neutral pH the binding of IFN-alpha A is 3-4 times less. This difference at neutral pH is seen on intact cells as well as on membrane preparations. The specific activity of IFN-alpha B is close to that of IFN-alpha A, both of which are 10-20 times more active than IFN-alpha D; and the binding titration is, therefore, independent of the initial binding affinities. 3. Using hybrid IFNs constructed from the DNA sequences of alpha D and alpha B, we have isolated the sequence responsible for the binding titration to the segment comprising amino acids 61-92. Examination of these sequences reveals that Lys-84 is present in all the IFN-alpha except IFN-alpha B where it is replaced by Glu; and Tyr-90, present in most of the common IFN-alpha including alpha A and alpha D, is replaced by Asp in IFN-alpha B. Lys and Tyr would normally titrate in the pH range 6-9. We conclude that the binding titration is due to an electrostatic interaction and we propose that the interaction is between IFN-receptor complexes. The role of the interaction in the binding losses that accompany the antiproliferative effects of IFN is discussed. PMID- 2964369 TI - The Ca2+-pumping ATPase and the major substrates of the cGMP-dependent protein kinase in smooth muscle sarcolemma are distinct entities. AB - It has been proposed that the plasma membrane Ca2+ pump of smooth muscle tissues may be regulated by cGMP-dependent phosphorylation [Popescu, L. M., Panoiu, C., Hinescu, M. & Nutu, O. (1985) Eur. J. Pharmacol. 107, 393-394; Furukawa, K. & Nakamura, H. (1987) J. Biochem. (Tokyo) 101, 287-290]. This hypothesis has been tested on a smooth muscle sarcolemma preparation from pig thoracic aorta. The actomyosin-extracted membranes showed ATP-dependent Ca2+ uptake as well as cGMP dependent protein kinase (G-kinase) activity. The molecular masses of the major protein substrates of the G-kinase (G1) and that of the Ca2+ pump were compared. Electrophoretic analysis of the phosphorylated intermediate of the sarcolemmal Ca2+-ATPase and the G1 phosphoprotein showed that these two proteins are not identical. The results were confirmed by using a 125I-calmodulin overlay technique and an antibody against human erythrocyte Ca2+-ATPase. Ca2+-uptake experiments with prephosphorylated membrane vesicles were carried out to elucidate possible effects of cGMP-dependent phosphorylation of membrane proteins on the activity of the Ca2+ pump. The cGMP-dependent phosphorylation was found to be extremely sensitive to temperature leading to very low steady-state phosphorylation levels at 37 degrees C. The difficulty was overcome by ATP[gamma S], which produced full and stable thiophosphorylation of G1 during the Ca2+ uptake experiments at 37 degrees C. However, the cGMP-dependent thiophosphorylation failed to influence the Ca2+-uptake properties of sarcolemmal vesicles. The results show that the Ca2+ pump of smooth muscle plasma membrane is not a direct target of the cGMP-dependent protein kinase and is not regulated by the cGMP-dependent phosphorylation of membrane proteins. PMID- 2964370 TI - Success and complication rates of coronary angioplasty in patients with and without previous myocardial infarction. AB - The primary success rate and incidence of major complications have been retrospectively assessed in a consecutive series of 224 patients undergoing percutaneous transluminal coronary angioplasty (PTCA) in one centre. The patients have been divided into three groups; those with angina and no previous myocardial infarction (Group 1; N = 130), those with angina and a previous transmural myocardial infarction (TMI) (Group 2; N = 59), and those with angina and a previous non-transmural myocardial infarction (NTMI) (Group 3; N = 26). The three groups were well matched for age, gender and angiographic severity of stenosis. The primary success rate in Group 1 was 90% compared to 64% in Group 2. The success rate in Group 3 lay in between at 77%. The lower success rates in Groups 2 and 3 were mainly due to an increase in the frequency of major complications. Acute coronary occlusion occurred in seven patients in Group 1, nine patients in Group 2 and four patients in Group 3. In all these patients in Groups 2 and 3 the outcome of acute occlusion was a procedure-related clinical myocardial infarction despite immediate re-angioplasty and/or emergency coronary artery bypass grafting whereas only four patients in Group 1 sustained an acute infarct. In this series of patients undergoing coronary angioplasty for symptom limiting angina, previous myocardial infarction appears to be a risk factor for a lower success rate mainly due to an increase in the frequency and severity of major complications. PMID- 2964371 TI - Expression of Fc gamma receptors on splenic T cells of mice infected with Plasmodium chabaudi adami. AB - In previous studies, it was shown that mice infected with Plasmodium chabaudi adami have a deficiency in their production of IgG1 immunoglobulin, suggesting isotype-specific immunoregulation. In order to examine this phenomenon in further detail the expression of Fc gamma receptors (Fc gamma R) on T cells obtained from mice infected with P. chabaudi was studied by flow cytometry. There was an increase in the number of splenic T cells which expressed Fc gamma R during infection. At the peak of the acute stage of infection (10-15 days) up to 40% of T cells were positive for Fc gamma R expression. These Fc gamma R were present on about 40% of both Lyt-2+ and L3T4+T cells. The isotype preference of these receptors on control Thy-1+ T cells is IgG1 greater than IgG2b greater than IgG2a as determined by an inhibition assay and fluorescence-activated cell sorter (FACS) analysis. However, 2 to 3 weeks after infection this pattern was altered such that IgG2b and IgG2a represented the major isotypes binding to the Fc gamma R of the L3T4+ T cell. At this stage of infection Fc gamma R on L3T4+ cells fail to bind IgG1. In the Lyt-2% T cells IgG1 and IgG2b remained the best inhibitors. These data suggest that there may be changes in Fc gamma R expression on T cells during infection reflected particularly in a decreased ability of IgG1 to bind to the Fc gamma R of L3T4+ cells. PMID- 2964372 TI - Anti-CD3 antibodies induce T helper function for human B cell differentiation in vitro by an interleukin 2-independent pathway. AB - Previously we have shown that interleukin 2 (IL 2) is an essential mediator in T cell-dependent B cell differentiation induced by pokeweed mitogen. Here we show that activation with monoclonal anti-CD3 antibodies of peripheral blood T cells led to the induction of helper activity for IgM secretion by human B cells from a prolymphatic leukemia. With the use of monoclonal antibodies against the IL 2 receptor and CD3+CD4+CD8- chronic lymphatic leukemia T cells with a strongly reduced capacity to produce IL 2, it was demonstrated that the anti-CD3-driven Ig secretion was obtained by an IL 2-independent pathway. The T cell help in this system is mediated by soluble factors. PMID- 2964373 TI - Regulatory activity of the human CD8+ cell subset: a comparison of CD8+ cells from the intestinal lamina propria and blood. AB - This study was done to better define the immunoregulatory mechanisms in the human intestinal lamina propria (LP). Peripheral blood (PB) and LP cells were obtained from patients having intestinal resections. CD4+ and CD8+ cell subsets were isolated using hybridoma antibodies in a panning technique. Graded numbers of LP and PB CD8+ cells were added to cultures of autologous fresh B cells plus CD4+ cells plus pokeweed mitogen. After 10 days incubation in vitro, the supernatants were collected, and IgM and IgA synthesis was measured by isotype-specific sandwich ELISA. Both PB and LP CD8+ cells suppressed IgM and IgA synthesis by indicator cultures consisting of 5 X 10(4) B cells plus 5 X 10(4) CD4+ cells to a comparable extent. However, when these same CD8+ cells were added to indicator cultures of 5 X 10(4) B cells plus 10(5) CD4+ cells, PB CD8+ cells still suppressed, but LP CD8+ cells enhanced IgM and IgA synthesis. LP but not PB CD8+ cells also augmented IgM and IgA synthesis in cultures with suboptimal immunoglobulin synthesis. Despite these results, LP CD8+ cells were not able to provide help for B cell immunoglobulin synthesis when these two cell types were cultured together with pokeweed mitogen. The mechanism of immunoglobulin augmentation by LP CD8+ cells appeared to involve antagonism of a CD4+ rather than CD8+ suppressor cells. We conclude that functional heterogeneity is more evident within the LP CD8+ subset, with both suppressor and contra-suppressor activities demonstrable, with the latter representing a major activity in LP but not in PB CD8+ cells. PMID- 2964374 TI - Enhancement in Igha mouse strains of the "natural" suppressive activity of normal T splenocytes against the expression of Igh-1b allotype. I. Molecular aspects of the chronic suppression obtained. AB - Several approaches have been used in our attempts to increase the "natural" ability of normal T splenocytes (Tn) from BALB/c or BC8 mice (both Igha) to induce, in F1 hybrids, a suppression of Igh-1b expression (IgG2a of b haplotype). These heterozygous F1 were produced by mating these Igha mice and their Ighb congenic partners (CB20 and C57BL/6, respectively). The most powerful approaches were to sensitize the Igha mice by either autologous splenocytes coated with Igh 1b or B splenocytes from Ighb-congenic mice. In F1 having paternally inherited the b haplotype the sensitized T splenocytes (Ts) prepared from such mice are able to induce, like Tn, when injected at birth, a chronic suppression of Igh-1b expression. However, the suppression was established with a much higher efficiency: already at 6 weeks of age in 100% of the F1 treated with 1 x 10(7) Ts, vs. a final rate of 70% progressively reached only at 42 weeks of age in the F1 treated with 4 x 10(7) Tn. In F1 having maternally inherited Ighb the differences were even more pronounced than with 4 x 10(7) Tn, i.e. the suppression induction was almost totally ineffective, whereas with 2 x 10(7)-4 x 10(7) Ts, a rate of 100% treated F1 subjected to suppression was reached at 19 weeks of age. As the productions of IgM, IgD and IgA of the b haplotype were not affected by the suppression, the Ts are believed to act on the Igh-1b+ cells. Attempts were also made to induce allotypic suppression of other b allotypes by the use, as sensitizing cells, of myeloma cells carrying Igh-6b (IgM of b haplotype). We failed in revealing any sign of a T cell reactivity against Igh-6b similar to the reactivity against Igh-1b. The use of Igh-6b+ myeloma cells grown in an Ighb or in an Igha background allowed us to assume that the cells responsible for the sensitization are, in the Ighb B lymphocyte population, either the Igh-1b+ lymphocytes or the lymphocytes having passively adsorbed this allotype, or both. PMID- 2964375 TI - A 5-HT1-like receptor mediates a sympathetic ganglionic hyperpolarization. AB - 5-HT induced a hyperpolarization of the rat superior cervical ganglion in vitro which was resistant to both MDL 72222 (10 microM), a 5-HT3 antagonist, and ketanserin (1 microM), a 5-HT2 antagonist. The 5-HT1-selective ligands 5 carboxamidotryptamine and 8-OH-DPAT also hyperpolarized the ganglion. The 5-HT induced hyperpolarization was potently antagonised by spiperone. These results suggest that 5-HT hyperpolarizes the rat superior cervical ganglion via a 5-HT1 like receptor which resembles the central 5-HT1A binding site. PMID- 2964376 TI - Rapid phosphorylation of MAP-2-related cytoplasmic and nuclear Mr 300,000 protein by serine kinases after growth stimulation in quiescent cells. AB - Antibody against brain microtubule-associated protein 2 (MAP-2) immunoprecipitated Mr 300,000 and 80,000 proteins of cultured fibroblasts and kidney cells. These proteins were not appreciably phosphorylated in quiescent cells, but were rapidly phosphorylated after growth stimulation by insulin, epidermal and fibroblast growth factors, transferrin, phorbol ester and diacylglycerol in the presence of Ca2+, in a manner similar to that of MAP-1 related Mr 350,000 protein (J. Cell Biol. 100, 748-753). A Ca2+ ionophore, which is known to make the quiescent cell competent but not to enter into the growth cycle, did not induce the phosphorylation. In a chase experiment, decay half lives of labeled phosphoproteins were 5 h for Mr 350,000 and 300,000 proteins, and 1.5 h for Mr 80,000 protein. On subcellular fractionation, phosphorylated Mr 350,000 and 300,000 proteins were detected first mainly in the cytoplasm and then in the nucleus, while Mr 80,000 phosphoprotein was consistently detected in the cytoplasm. The phosphorylation of these proteins occurred on serine residues after stimulation with various factors. Thus, the phosphorylation of cytoskeleton associated Mr 350,000 and 300,000 proteins by serine kinases seems to be a common second process after growth stimulation and to link cytoplasmic and intranuclear events. PMID- 2964378 TI - Influence of fibroblast growth factor on phosphorylation and activity of a 34 kDa lipocortin-like protein in bovine epithelial lens cells. AB - We examined the effect of basic fibroblast growth factor (FGF) on phosphorylation and lipocortin-like activity of the 34 kDa protein present in the basal membrane of epithelial peripheral cells which initiate growth and differentiation in the bovine lens. We found that: (i) the 34 kDa protein possesses anti-phospholipase A2 (PLA2) activity of lipocortin; (ii) in response to FGF, the anti-PLA2 activity of this protein is enhanced whereas its phosphorylation is markedly decreased. It is suggested that the 34 kDa protein might represent an important biological activity in controlling FGF induction of growth and differentiation in the adult eye lens. PMID- 2964377 TI - Studies on the serotonin transporter in platelets. AB - [3H]-Imipramine and [3H]-paroxetine label with high affinity a recognition site which is associated with the serotonergic transporter in blood platelets. The pharmacological profile of [3H]-imipramine and [3H]-paroxetine binding is highly correlated with the potency of drugs to inhibit the uptake of serotonin. Dissociation kinetic experiments suggest that the substrate recognition site for serotonin may be different from the modulatory site which is labeled with [3H] imipramine or [3H]-paroxetine. The existence of an endocoid acting on the imipramine receptor to modulate the serotonin transporter has been proposed by several laboratories. In clinical studies most laboratories have reported a decrease in Bmax of [3H]-imipramine binding in platelets from depressed untreated patients when compared with matched healthy volunteers. The Bmax of [3H] imipramine binding in platelets appears to be a state-dependent biological marker in depression. PMID- 2964379 TI - Two-chain structure of the interleukin 1 receptor. AB - By crosslinking radioiodinated recombinant human IL1 alpha to mouse EL4 thymoma cells we have identified in addition to the known IL1-binding proteins of 80 kDa, a second IL1-binding protein of about 40 kDa. This second binding protein could be demonstrated most easily when crosslinking to higher protein complexes was inhibited. This finding suggests that the IL1 receptor, similar to the receptor for other cytokines such as interleukin 2, is composed of a heterodimer, of which both polypeptides contribute to ligand binding. PMID- 2964380 TI - Regulation of fructose 2,6-bisphosphate levels in cold-acclimated brown adipose tissue of rat. AB - The effects of cold exposure and T4 administration on fructose 2,6-bisphosphate levels, phosphofructokinase-2 and pyruvate kinase activities were examined in rat brown adipose tissue. Cold adaptation (14 days) gave rise to a 2-fold increase in the amount of fructose 2,6-bisphosphate and phosphofructokinase-2 activity, and increased the pyruvate kinase activity 4-fold. If, in addition, the cold acclimated rats were treated with T4, these parameters were again significantly enhanced. The effect on phosphofructokinase-2 was on the Vmax, without modification of the Km (for both fructose 6-phosphate and ATP) of the enzyme. In the hypothyroid state, however, the activity of pyruvate kinase remains unchanged. These data support previous observations on stimulation of glycolytic flux during cold adaptation in brown adipose tissue, and a permissive role of thyroid hormones in the process. PMID- 2964381 TI - 125I-d(CH2)5[Tyr(Me)2, Tyr(NH2)9]AVP: iodination and binding characteristics of a vasopressin receptor ligand. AB - A radioiodinated vasopressin antagonist, d(CH2)5[Tyr(NH2)9]AVP has been prepared. Iodination was carried out at the phenyl moiety of the tyrosylamide residue at position 9, followed by HPLC purification. Non-radiolabelled monoiodinated antagonist was used as a reference for identification. 125I-d(CH2)5[Tyr(Me)2, Tyr(NH2)9]AVP binding appeared to take place with a dissociation constant of 0.28 +/- 0.09 nM (Kd +/- SD) to V1 vasopressin receptors on rat liver membranes. PMID- 2964382 TI - Laparoscopy for pancreatic cancer: does it benefit the patient? AB - The findings in 73 patients undergoing laparoscopy for pancreatic cancer have been reviewed. The procedure was performed immediately prior to a proposed laparotomy which was undertaken in 51 patients. In this subgroup, 42 patients were correctly staged as having incurable/inoperable disease by laparoscopy but only four out of nine patients judged to be resectable were found to be operable at laparotomy. Laparoscopic target biopsy/fine needle cytology of the primary or its secondary deposits confirmed the diagnosis of pancreatic cancer in 61 out of 65 patients (92%). Whereas the majority of hepatic deposits visualized by laparoscopy had been detected by the prelaparoscopic imaging tests, peritoneal and omental deposits were only identified by laparoscopy. Laparoscopy is a useful procedure in the staging of patients with pancreatic cancer and establishes the diagnosis of advanced disease thereby, obviating a laparotomy in all those patients in whom surgical palliation is not indicated. The procedure should be performed in those patients in whom a laparotomy is contemplated. PMID- 2964383 TI - The properties of a glass polyalkenoate (ionomer) cement incorporating sintered metallic particles. PMID- 2964384 TI - [Genetic polymorphism of the alpha 1-antitrypsin system in the native population of the Kazakh SSR]. AB - Phenotypes of alpha 1-antitrypsin of 218 natives from three ethno-historical regions of the Kazakh SSR were detected by isoelectric focusing in ultrathin layer polyacrylamide gel. Gene frequencies of the PiM1, PiM2, PiM3 subtypes in the summary extract were as follows: 0.8477, 0.1372 and 0.0106, respectively; the total gene frequency of two rare variants (PiN and PiZ) was 0.0046. The observed distribution of Pi subtypes shows good agreement with the Hardi-Weinberg equation. The analysis of the interpopulation intraethnic variability of the alpha 1-antitrypsin phenotype and allele frequencies in Kazakhs revealed clear local diversity. The frequency of PiM1 in the natives from North-Central ethno historical region was reliably lower and that of PiM2 higher than in populations from South-East and West regions. The extracts analysed did not differ in the PiM3 incidences. The results of these studies were compared with the literature data for alpha 1-antitrypsin polymorphism in populations of the Euro-Asia. It is shown that PiM1 and PiM2 frequencies in the Kazakhs differ from the corresponding mean values both in mongoloid and europeoid groups. At the same time, they do not correspond to the intermediate frequency estimations, which could be expected from the fact of mixed origin of the Kazakh people and their border-line geographical position between Europe and Asia. Possible reason for such discrepancy is discussed. PMID- 2964385 TI - Construction of an ordered overlapping library of bacteriophage P1 DNA in phage vector lambda D69. AB - A library of bacteriophage P1 DNA was constructed in the phage vector lambda D69. The DNA of some 150 randomly chosen lambda-P1 hybrid phages containing P1 DNA fragments 5-10 kb in size was analyzed by restriction endonuclease digestion using enzymes EcoRI, BglII, and BamHI that cleave P1 DNA at known positions on the physical map of P1. Approximately one third of the phages contained P1 DNA inserts having two or more restriction sites for any one of these enzymes, thus allowing the location of the insert to be determined with respect to the physical map. Genetic tests allowed detection of lambda-P1 hybrid phages possessing inserts with functional P1 ban and CmR genes. A subset of 18 phages was analyzed in more detail; their P1 DNA inserts comprise an ordered collection of overlapping P1 DNA fragments that cover almost 98% of the P1 genome. PMID- 2964386 TI - Molecular cloning and sequencing of sucrose synthase cDNA from potato (Solanum tuberosum L.): preliminary characterization of sucrose synthase mRNA distribution. AB - A cDNA library constructed from poly(A)+ RNA from potato tuber in lambda gt11 was screened for sucrose synthase using a maize sucrose synthase cDNA probe. The longest 2.7-kb insert was sequenced. An ATG located within the sequence CTGCAATGG starts an open reading frame of 805 codons. The nucleotide sequence, when compared to the maize sucrose synthase cDNA sequence, exhibits about 70% identity and the deduced amino acid sequence about 75%. Three amino acid regions are about 90% homologous and two of them could be important for the protein function. Expression studies show that transcription of the potato sucrose synthase gene is at least ten fold higher in tubers compared to photosynthetically active tissues. PMID- 2964387 TI - Effect of litter size (i.e. nutrition) on carbon monoxide-induced persistent heart changes. AB - It was hypothesized that manipulation of litter size, thus nutrition, which has been shown to alter the neonatal response to cardiovascular stress (i.e. carbon monoxide) might also alter persistent post-stress changes. Rat pups reared in litters of 4 and 16 inhaled 500 ppm CO for 32 days ("CO"), or served as controls ("AIR"). Some pups were killed at 14-15 days of age to assess initial cardiomegaly. As adults, right ventricle (RV) mass was greater in CO/4 males (123 days of age) and females (113 days of age) than in same sex CO/16 rats. Persistent RV cardiomegaly was present in CO/4 males and females compared to AIR's of the same litter size and sex, whereas this was the case only in females comparing CO/16 and AIR/16 rats. RV mass was significantly larger in AIR/4 males than in AIR/16 males. Plots of initial cardiomegaly (both ventricles) versus persistent cardiomegaly (RV) for large and small litters produced similar slopes for the two sexes, with females lying above males. Resting heart rate, monitored in males (66-121 days of age) and females (81-109 days of age), was increased by small litter size, and also by CO, particularly in the males. Resting heart rate was significantly correlated with RV weight. RV DNA content and concentration were increased by small litter size in the males, and concentration also by CO. The male CO/4 rats had the highest DNA content and concentration. In the females, DNA content was increased by small litter size and was greatest in the CO/4 group. Thus, the effects of small litter size have lasting effects: i.e. augmented persistent cardiomegaly, persistent tachycardia, and myocardial DNA content and concentration. PMID- 2964388 TI - Continuous infusion fluoropyrimidines as salvage therapy for patients with advanced ovarian carcinoma. AB - A major challenge facing those caring for patients with ovarian carcinoma is inducing remission following the failure of first-line treatment. Toward this end, we have examined the efficacy of continuous infusion fluoropyrimidines. During a 2-year period, nine patients with recurrent ovarian carcinoma received treatment with continuous infusion 5-fluorouracil (5-FU) or 5-fluorouracil deoxyribose as single agents or in combination with other drugs. Eight patients were evaluable, with responses obtained with each treatment regimen. Myelotoxicity was mild, with only 3 episodes of grade 4 toxicity out of 70 total cycles of chemotherapy. Mucositis was moderate to severe, tending to be the dose limiting adverse effect. Continuous infusion 5-FU in these combinations appears to be active with very acceptable toxicity in heavily pretreated patients. PMID- 2964389 TI - Sclerosing stromal tumor of the ovary associated with endometrial adenocarcinoma: a case report. AB - The first case of ovarian sclerosing stromal tumor associated with endometrial adenocarcinoma in a 44-year-old woman is presented. By ultrastructural study, typical steroid-secreting type theca cells and various transitional tumor cells from fibroblast-like cells to theca-like cells were observed in the ovarian tumor. The clinical history of the patient and the ultrastructural study of this tumor suggested hormonal activity, but no apparent evidence of active hormone secretion was detected by plasma analysis for steroid hormones. PMID- 2964390 TI - [Acne vulgaris: pathogenesis and treatment]. PMID- 2964392 TI - [The iliolumbar syndrome]. PMID- 2964391 TI - [Thermocoagulation of lumbosacral facets for mechanical low-back pain syndrome]. PMID- 2964393 TI - Successful foster caregivers of geriatric patients. AB - The authors examined the characteristics of caregivers from two foster care programs designed to substitute for nursing home placement of severely disabled elders. The more successful caregivers became foster families for humanitarian reasons, had more caregiving experience, gained satisfaction from the role, involved the patient in family activities, and did not feel more religious than other people. The findings provide guides for screening applicant foster caregivers of geriatric patients. PMID- 2964394 TI - Out-of-hospital health care. PMID- 2964395 TI - Planning for incapacity: two perspectives on safeguards. PMID- 2964396 TI - A comparative study of plasma vasopressin levels and V1 and V2 vasopressin receptor properties in congenital hypothyroid rat under thyroxine or vasopressin therapy. AB - The effects of propylthiouracil (PTU) treatment on the plasma vasopressin level, on the number of hepatic (V1) or renal (V2) vasopressin receptors and on the hormone-sensitive adenylate cyclase activity in the kidney of developing rats were studied in parallel. In addition, we investigated the corrective effects of thyroxine therapy on the plasma vasopressin level and parameters related to the liver, and the effects of vasopressin therapy on the parameters related to the kidney. As already reported in the case of the number of V2 receptors and adenylate cyclase activity in the kidney, the deficient plasma vasopressin level in hypothyroid rats was completely corrected by two daily physiological doses of thyroxine given from birth to the age of sacrifice (1 month). Unlike the V1 receptors, the V2 receptors are known to be highly dependent on their specific circulating ligand. Since, first of all, the deficit was similar in the numbers of V1 and V2 receptors in hypothyroid rats, and, secondly, the treatment of hypothyroid rats by two daily physiological doses of long lasting vasopressin was found ineffective to recover the deficit in the number of V2 receptors, it can be concluded that thyroid deficiency directly alters vasopressin receptor biosynthesis in both liver and kidney, instead of acting via the depressed plasma vasopressin level. PMID- 2964397 TI - Heat resistance, immunological and quantitative changes of neutrophil alkaline phosphatase in trisomy 21 pregnancies. AB - Neutrophil alkaline phosphatase (NAP) was analysed in 25 pregnant women with trisomy 21 foetuses whose chromosomal aberration was recognized by cytogenetic study after amniocentesis. Enzyme investigation was performed at 20-22 weeks of gestation using cytochemical and biochemical techniques. Twenty-nine women at the same stage of normal pregnancies were selected as controls. In parallel, each mother was karyotyped. Ten subjects from each series underwent biochemical and immunological investigation: measurement of enzyme levels, thermostability study and immunological tests with alkaline phosphatase isoenzyme antibodies. NAP from pregnant women with trisomy 21 foetuses was characterized by: (1) a lower rate of enzyme activity, (2) a large amount of heat-stable enzyme (T = 56 degrees C for biochemical assays, T = 85 degrees C for cytochemical tests), and (3) a marked loss of liver antigenicity. These findings suggest the presence in trisomy 21 pregnancies of a non-specific alkaline phosphatase isoenzyme which appears as an "enzyme marker" in maternal circulating neutrophils. PMID- 2964398 TI - Parental centromere separation sequence and aneuploidy in the offspring. AB - We have studied the centromere separation sequence in lymphocyte mitoses of the parents of four infants with trisomy 18, five patients with trisomy 21, and five children with normal karyotype. "Late separation" of chromosome 18 was found in both parents of a neonate with trisomy 18, "early separation" of chromosome 21 in three mothers and in one father of four children with trisomy 21. No "out-of phase" separation occurred in the mitoses of the parents of normal children. The findings provide further evidence for the correlation between alteration of the parental centromere separation sequence and aneuploidy of the offspring. PMID- 2964399 TI - Nucleolus organizer region(s) activity and acrocentric chromosome association in Down syndrome. PMID- 2964400 TI - The heart in hypertension: unresolved conceptual challenges. Special lecture. AB - Much has been learned over the past 25 years concerning the role of the heart in hypertension. In a multiplicity of areas a great deal has been clarified but a number of issues remain unresolved. This personal overview outlines some of these challenging areas for investigation, including questions relating to the cardiogenic reflexes, mechanisms underlying total body autoregulation that may involve not only the adaptation of arterioles but also venoconstriction in hypertension, postcapillary constriction also involving the efferent glomerular arterioles, the mechanisms underlying the development and regression of hypertrophy as well as the function of the hypertrophied and "regressed hypertrophy" heart, and the precise hemodynamic actions of atrial natriuretic factor. PMID- 2964401 TI - Atrial natriuretic factor in essential hypertension and adrenal disorders. AB - Patients with untreated essential hypertension had significantly higher plasma atrial natriuretic factor (ANF) levels (92.9 +/- 12.9 pg/ml, mean +/- SE) than those of age-matched controls (37.8 +/- 6.0 pg/ml; p less than 0.01). Plasma ANF levels in essential hypertensive patients showed a significant positive correlation with mean arterial pressure (MAP; r = 0.46, p less than 0.05) and an inverse correlation with plasma renin activity (PRA; r = -0.43, p less than 0.05). Plasma ANF levels after medication showed significant correlation with the decrease in MAP (r = 0.565, p less than 0.05). Patients with primary aldosteronism had significantly higher plasma ANF levels (122.4 +/- 30.2 pg/ml, n = 8) than those of controls (p less than 0.05). The levels returned to normal after extirpation of adrenal tumors. The response of plasma ANF levels in patients with primary aldosteronism to volume expansion with infusion of 2 L of physiological saline in 2 hours was greater than in controls. Such exaggerated response disappeared after surgical treatment. Infusion of angiotensin II (Ang II; 20 ng/kg/min) or norepinephrine (200 ng/kg/min) for 30 minutes to normal volunteers (n = 5) resulted in a rise in MAP (24.9 +/- 3.3 and 15.8 +/- 4.4 mm Hg, respectively) and a twofold increase in plasma ANF level. Infusion of the Ang II antagonist [Sar1, Ile8]Ang II (600 ng/kg/min) for 30 minutes, resulted in a rise in MAP (18.8 +/- 2.1 mm Hg) and more than a twofold increase in plasma ANF level in patients with essential hypertension (n = 6).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964402 TI - The atrial natriuretic factor in hypertension. State of the art lecture. AB - Studies were conducted to assess the effects of bolus injections and infusions of human atrial natriuretic factor (ANF) in control subjects and patients with mild essential hypertension, and to measure plasma immunoreactive ANF (irANF) concentration in a large group of patients with essential hypertension. The results are compared with those obtained by other groups on the measurements of plasma irANF in hypertensive patients. It appears that plasma irANF concentrations are not increased in patients with mild essential hypertension despite the evidence of increased preload and of atrial distention as reported by others. This suggests a hyporesponsiveness of the atria to release ANF. PMID- 2964403 TI - Ganglionic immunoreactive atrial natriuretic factor in rat experimental hypertension. AB - Because previous data have suggested a dependence of ganglionic atrial natriuretic factor (ANF) content on preganglionic cholinergic input, we investigated the possibility that the increased neural activity observed in spontaneously hypertensive rats (SHR) may be reflected by ganglionic immunoreactive ANF levels. Four-week-old normotensive SHR had celiac ganglionic immunoreactive ANF values comparable to those of Wistar-Kyoto rats (WKY). When they became hypertensive, however, at 12 weeks of age, the SHR manifested higher immunoreactive ANF levels in celiac ganglia than the WKY group (25.3 +/- 2.6 vs 14.5 +/- 1.7 pg/ganglion; p less than 0.01), but there were no differences in levels in the superior cervical and nodose ganglia. The values in celiac ganglia were quadrupled on the average in hypertensive Dahl salt-sensitive rats under the influence of an 8% salt intake for 5 weeks, but no difference was noted in any of these ganglia between this group and their salt-resistant partners. The celiac and superior cervical ganglionic immunoreactive ANF content in normotensive Sprague-Dawley rats was higher with high salt than with normal salt intake. Hypertensive rats treated with deoxycorticosterone acetate (DOCA)-salt and sham treated controls showed immunoreactive ANF concentrations in celiac ganglia similar to those detected in Dahl rats but, again, no differences were found between groups. Thus, hypertensive SHR, compared to WKY, have higher celiac ganglionic immunoreactive ANF levels, unlike Dahl salt-sensitive and DOCA-salt animals relative to their respective controls. This increase is unique to SHR (although all three models have elevated plasma immunoreactive ANF when they are hypertensive) and to the celiac ganglia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964404 TI - Secretion of N-terminal fragment of gamma-human atrial natriuretic polypeptide. AB - To elucidate the posttranslational processing of gamma-human atrial natriuretic polypeptide (human atrial natriuretic factor-[1-126]), which is a prohormone of alpha-human atrial natriuretic polypeptide (human atrial natriuretic factor-[99 126]), and the secretion of gamma-human atrial natriuretic polypeptide-derived peptides from the heart, we established a radioimmunoassay specific for the N terminal sequence of gamma-human atrial natriuretic polypeptide, gamma-human atrial natriuretic polypeptide-(1-25), as well as a radioimmunoassay for alpha human atrial natriuretic polypeptide. With the aid of the radioimmunoassays for gamma-human atrial natriuretic polypeptide-(1-25) and for alpha-human atrial natriuretic polypeptide, we detected 290 +/- 35.6 pg/ml of gamma-human atrial natriuretic polypeptide-(1-25)-like immunoreactivity in plasma from healthy humans, while the simultaneously determined plasma alpha-human atrial natriuretic polypeptide-like immunoreactivity level was 20.9 +/- 2.8 pg/ml. Correlation between the two values was significant. High performance gel permeation chromatographic analysis revealed that the plasma gamma-human atrial natriuretic polypeptide-(1-25)-like immunoreactivity was composed of a component (molecular weight, 10,000) without alpha-human atrial natriuretic polypeptide-like immunoreactivity, while the plasma alpha-human atrial natriuretic polypeptide like immunoreactivity was composed of alpha-human atrial natriuretic polypeptide with a molecular weight of 3000. In patients with heart diseases, the plasma gamma-human atrial natriuretic polypeptide-(1-25)-like immunoreactivity level showed a concomitant and graded increase, with the plasma alpha-human atrial natriuretic polypeptide-like immunoreactivity level in agreement with the severity of the disease. There were significant positive correlations between the two immunoreactivity levels and right or left atrial pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964405 TI - Brain renin-angiotensin. Central control of secretion of atrial natriuretic factor from the heart. AB - To elucidate the modulatory role of the brain renin-angiotensin system in the regulation of the secretion of atrial natriuretic factor (ANF) from the heart, the effects of intracerebroventricular administration of angiotensin II on the plasma ANF level were examined in conscious unrestrained rats. Administration of angiotensin II in doses of 100 ng and 1 microgram significantly enhanced ANF secretion induced by volume loading with infusion of 3 ml of saline (peak values of the plasma ANF level: control, 220 +/- 57 pg/ml; angiotensin II 100 ng, 1110 +/- 320 pg/ml, p less than 0.01; angiotensin II 1 microgram, 1055 +/- 60 pg/ml, p less than 0.01). Injection of angiotensin II alone had no significant effect on the basal plasma ANF level. Central angiotensin II-induced ANF secretion was significantly attenuated by pretreatment with intravenous administration of the V1-receptor antagonist of vasopressin or intracerebroventricular administration of phentolamine. These results indicate that the brain renin-angiotensin system modulates ANF secretion in response to volume loading through the stimulation of vasopressin secretion or the activation of the central alpha-adrenergic neural pathway. PMID- 2964406 TI - Atrial natriuretic factor immunoreactivity in human fetal lung tissue and perfusates. AB - Immunoreactive atrial natriuretic factor (ANF) was detected in human fetal homogenates and perfusates using a sensitive and specific radioimmunoassay for the 28 amino acid (C-terminal) fragment. Three peaks of ANF immunoreactive material were found in the lung homogenates. With high performance liquid chromatography, the elution characteristics of the first immunoreactive peak were the same as those of circulating human ANF. The other two peaks have not been characterized, although one had a position similar to the 126 amino acid rat prohormone (Asn 1-Ile 110-Tyr 126). The time course of release of immunoreactive ANF by perfused human fetal lungs was also studied. It is suggested that ANF may play a role in early pulmonary function. PMID- 2964407 TI - Pindolol, not propranolol, reverses cardiac hypertrophy in renal hypertensive rats. AB - Reversal of cardiac hypertrophy has been obtained by treatment with some antihypertensive drugs but has not been achieved consistently with beta blockers. To investigate whether this difference might be explained by the distinct hemodynamic actions of the drugs, we studied the effects of propranolol and pindolol, beta blockers with distinct modes of action, on cardiac hypertrophy of hypertensive male Wistar rats, two-kidney, one clip (2K1C) Goldblatt model (n = 33) and sham-operated control rats (n = 34). We also assessed the effects of such therapies on the ventricular pumping ability during open-chest, transient aortic occlusion. Four weeks after surgery, propranolol (5 mg/kg/day p.o.) was given to hypertensive (n = 8) and control rats (n = 11); pindolol was also given orally (1 mg/kg/day) to similar groups (n = 7 and n = 5, respectively). Untreated animals served as controls for both groups. Cardiac hypertrophy developed with hypertension in the untreated rats of the propranolol (3.38 +/- 0.18 vs 2.60 +/- 0.08 mg/g; p less than 0.01) and pindolol groups (3.93 +/- 0.21 vs 2.40 +/- 0.03 mg/g; p less than 0.001). Treatment reversed cardiac hypertrophy in the pindolol treated (3.01 +/- 0.19 vs 3.93 +/- 0.21 mg/g; p less than 0.001, NS) but not in the propranolol-treated rats (3.24 +/- 0.18 vs 3.38 +/- 0.21 mg/g, NS). The maximal pressure that developed during aortic occlusion in the propranolol group was similar to that observed in the pindolol group. These results indicate that cardiac hypertrophy is reversed by pindolol but not by propranolol, and that this reversal does not interfere with left ventricular pumping ability. PMID- 2964408 TI - Sex hormone modulation of ventricular hypertrophy in sinoaortic denervated rats. AB - The influence of sex hormones on the development of left ventricular hypertrophy was investigated in baroreceptor-denervated rats. A significant increase (p less than 0.01) in the left ventricular weight/body weight ratio was observed in male but not in female rats 15 days after operation, compared to age- and sex-matched sham-operated rats. This differential hypertrophy occurred despite the development of a significant elevation in arterial blood pressure in both sexes. Castration prior to sinoaortic denervation did not change the level of arterial hypertension but caused a significant reduction (p less than 0.01) in left ventricular weight in male rats and a significant increase (p less than 0.01) in female rats. The pretreatment of male and female sinoaortic denervated and castrated rats with testosterone resulted in ventricular hypertrophy similar to that observed in intact male sinoaortic denervated rats. Pretreatment with estradiol, however, suppressed the left ventricular hypertrophy in intact male rats but did not change the normal ventricular mass observed in intact female sinoaortic denervated rats. These results indicate that the development of left ventricular hypertrophy in sinoaortic denervated rats is modulated by sex hormones, and that testosterone exerts a facilitatory and estradiol an inhibitory action. PMID- 2964409 TI - Counting the people disabled by head injury. AB - Figures for the incidence of head injury are frequently cited but there has been no accurate estimate and the number of individuals left with persistent disability is not exactly known. This study identified all those admitted to hospital with a diagnosis of head injury, in a known population. In 1 year there were 160 survivors of head injury per 100,000 of population. Rates of disability of 0.38 per 100,000 in need of care and 0.75 per 100,000 unemployable and primarily in need of occupation, were found, excluding those over 65 years old. PMID- 2964410 TI - Iron acquisition by Haemophilus influenzae. AB - The mechanisms for acquisition of iron by Haemophilus influenzae and their role in pathogenesis are not known. Heme and nonheme sources of iron were evaluated for their effect on growth of type b and nontypable strains of H. influenzae in an iron-restricted, defined medium. All 13 strains acquired iron from heme, hemoglobin, hemoglobin-haptoglobin, and heme-hemopexin. Among nonheme sources of protein-bound iron, growth of H. influenzae was enhanced by partially saturated human transferrin but not by lactoferrin or ferritin. Purified ferrienterochelin and ferridesferrioxamine failed to provide iron to H. influenzae, and the supernatants of H. influenzae E1a grown in iron-restricted medium failed to enhance iron-restricted growth of siderophore-dependent strains of Escherichia coli, Salmonella typhimurium, and Arthrobacter terregens. Marked alterations in the profile of outer membrane proteins of H. influenzae were observed when the level of free iron was varied between 1 microM and 1 mM. Catechols were not detected in the supernatants of strain E1a; however, iron-related hydroxamate production was detected by two biochemical assays. We conclude that the sources of iron for H. influenzae are diverse. The significance of hydroxamate production and iron-related outer membrane proteins to H. influenzae iron acquisition is not yet clear. PMID- 2964411 TI - Induction of antigen-specific T suppressor cells by soluble Paracoccidioides brasiliensis antigen. AB - In naturally acquired paracoccidioidomycosis, patients have depressed in vivo and in vitro cell-mediated immune (CMI) responses to Paracoccidioides brasiliensis antigen. In addition, it has been reported that these patients have significant levels of circulating paracoccidioidal antigen in their sera. The primary purpose of this investigation was to assess the effects of P. brasiliensis antigen on the CMI responses in a mouse model. On the basis of findings with other fungal agents, we predicted that circulating paracoccidioidal antigen may be inducing suppressor cells which modulate the CMI response. In this study, we show (i) that a soluble P. brasiliensis culture filtrate antigen (Pb.Ag) emulsified in complete Freund adjuvant and injected subcutaneously into mice induces reasonably high levels of delayed-type hypersensitivity (DTH) in CBA/J mice; (ii) that Pb.Ag elicits DTH reactions specific for P. brasiliensis when injected into footpads of immunized mice; and (iii) that an intravenous injection of Pb.Ag induces a population of lymph node and spleen cells which, upon adoptive transfer, suppress the afferent limb of the DTH response to paracoccidioidal antigen. The afferent suppressor cells can be detected in spleens as early as 5 days after Pb.Ag treatment, are present in significant numbers by 7 days in both spleens and lymph nodes, and are virtually absent by 14 days. In contrast, at 14 days after antigen injection, efferent suppressor cells were detected in spleens and lymph nodes. The Pb.Ag-induced afferent suppressor cells specifically inhibit the antiparacoccidioidal DTH response. They are nylon wool-nonadherent cells, and their activity is abrogated by anti-Thy-1 and complement treatment, indicating that they are T lymphocytes. The phenotype of these afferent suppressor T cells is L3T4+ Lyt-1+2- I-J+. The Pb.Ag-specific suppressor cells described in this paper are similar to the Ts1 cells in the azobenzenearsonate, 4-hydroxy-3 nitrophenyl acetyl, and cryptococcal models of suppression of the DTH response and to the afferent suppressor cells in the dinitrofluorobenzene contact sensitivity system. PMID- 2964412 TI - Characterization of efferent T suppressor cells induced by Paracoccidioides brasiliensis-specific afferent T suppressor cells. AB - Previously, we reported that Paracoccidioides brasiliensis culture filtrate antigen (Pb.Ag) when injected i.v. into mice induces antigen-specific suppressor cells which down-regulate the anti-P. brasiliensis delayed-type hypersensitivity (DTH) response. The suppressor cells are present in both spleens and lymph nodes of Pb.Ag-treated animals and suppress the afferent limb but not the efferent limb of the DTH response to P. brasiliensis. The suppressor cells induced by Pb.Ag are L3T4+ Lyt-1+2- I-J+ T cells and are considered to be equivalent to the Ts1 cells described for other antigen-specific suppressor cell pathways. This report provides data which show that Ts1 cells induced by Pb.Ag or a soluble factor derived from Ts1 cells (TsF1) stimulates the production of second-order or efferent suppressor cells. The second-order suppressor cells are detectable in spleens and lymph nodes of mice 7 days after injection of Ts1 cells or TsF1 and are specific in suppressing the paracoccidioidal DTH response. In addition, the second-order suppressor cells are T cells with an L3T4- Lyt-2+ I-J+ phenotype and are effective in suppressing only the efferent limb of the P. brasiliensis DTH response. On the basis of the characteristics defined in this study, the paracoccidioidal second-order suppressor cells are equivalent to the Ts2 cells described for other antigen-specific suppressor-cell pathways. Thus, the suppressive circuit induced by Pb.Ag is similar to the suppressor-cell pathways that regulate the DTH responses to azobenzenearsonate, 4-hydroxy-3-nitrophenyl acetyl, lysozyme, and Cryptococcus neoformans antigen. We propose that such a suppressor-cell circuit as defined here with the murine model could be responsible for the depressed cell-mediated immune responses observed in paracoccidioidomycosis patients who have antigen circulating in their sera. PMID- 2964413 TI - Isolation of a glucan-binding domain of glucosyltransferase (1,6-alpha-glucan synthase) from Streptococcus sobrinus. AB - A glucan-binding domain of 1,6-alpha-glucan synthase (dextransucrase) (GTF-S) was isolated from a trypsin digest of the Streptococcus sobrinus enzyme. The large 60.5-kilodalton peptide had an affinity for dextran comparable to that of the native enzyme, but had no glucan synthesis activity. The domain was produced in high yield compared with other large cleavage products, which allowed easy purification by size exclusion high-pressure liquid chromatography and affinity chromatography. Two other proteases (mouse submaxillaris protease and lysyl endopeptidase) with specificities similar to trypsin generated a distribution of GTF-S peptides that was also greatly enriched in the glucan-binding peptide. Proteases with markedly different specificities (chymotrypsin and Staphylococcus aureus V8 protease) produced a family of peptides with some evidence of the glucan-binding domain but in far lower yield. The tertiary structure of the domain was critical to its resistance to proteolysis; heat denaturation of GTF-S before trypsin digestion resulted in cleavage of the enzyme to small limit peptides leaving no evidence of the glucan-binding domain. The amino acid composition of the peptide was very similar to that of the native enzyme. The common occurrence of proteases in oral streptococcus cultures and reports of glucosyltransferase degradation during purification and storage raises the possibility that some accounts of glucan-binding receptors are peptides derived from glucosyltransferase. Kinetic implications of a glucan-binding domain are discussed. PMID- 2964414 TI - Back disorders in crane operators exposed to whole-body vibration. AB - In The Netherlands so far little research has been carried out to investigate the health effects of exposure to whole-body vibration at work. In a retrospective (10-year) follow-up study, the incidence of permanent work disabilities in crane operators exposed to vibration was compared to that of a control group. The emphasis in this presentation is on disablement because of back disorders. Among the crane operators with more than 5 years of exposure to vibration, more disability due to back trouble occurred than in the control group. The incidence density ratio (IDR) exceeded 2 for disability because of degeneration of the intervertebral disc, after adjustment for age, nationality, shift work and calendar year. With increasing years of exposure, the risk ratio for disability because of intervertebral disc disorders also increased. The risk ratio for a disability pension due to disease of the intervertebral disc for crane operators with more than 5 years of exposure was almost 3 compared to the control group. A combination of exposure to whole-body vibration, an unfavorable posture, and climatic conditions is responsible for the development of these health effects. PMID- 2964415 TI - Occupational skin disease in Finland. An analysis of 10 years of statistics from an occupational dermatology clinic. AB - An analysis of 10 years (1974-1983) of statistics was carried out at the Institute of Occupational Health, Helsinki, Section of Dermatology, which is devoted to occupational dermatology. A total of 1,082 cases of occupational skin diseases were diagnosed during this period. Allergic (50.1%) and toxic eczema (47.1%) comprised the majority of occupational cases of dermatosis. The most frequent causes of allergic occupational eczemas were rubber chemicals (19.9%), chromates (18.8%), and epoxy resins (13.1%). If the metals (chromium, nickel and cobalt) were considered as a group, they formed the largest category (28.4%), followed by the plastic materials (27.7%). Detergents (37.8%), followed by organic solvents (16.1%), were responsible for most of the irritant (toxic) eczemas. Occupational skin diseases currently make up about 20% of all occupational diseases in Finland, but the percentage is decreasing. PMID- 2964416 TI - Guidelines for antibiotic prophylaxis of infective endocarditis for dental patients with cardiovascular disease. Federation Dentaire Internationale Technical Report No. 28. PMID- 2964417 TI - Teicoplanin in patients with chronic renal failure on dialysis: microbiological and pharmacokinetic aspects. AB - A significant increase in the incidence of methicillin resistance was found in coagulase-negative staphylococci isolated from infected dialysis fluids in 1985 compared with the previous year. Vancomycin and teicoplanin were active against all these isolates, and had similar minimum inhibitory and bactericidal concentrations. The pharmacokinetic behaviour of teicoplanin in 23 dialysis patients was studied. A single, intravenous dose of teicoplanin was given to 11 patients on haemodialysis (HD) and seven patients on chronic ambulatory peritoneal dialysis (CAPD). In five CAPD patients, 40 mg was added to each 2 litre bag of dialysate for a five day period. Twenty such bags were exchanged. The study showed that a) teicoplanin was not removed from the body by HD or CAPD, b) less than 3% of the administered dose appeared in the urine, c) serum levels reached a plateau of 3-4 micrograms/ml after 40 hours and were maintained for at least five days, regardless of the route of administration or form of dialysis. These findings have obvious implications regarding appropriate treatment regimens in dialysis patients. PMID- 2964418 TI - Teicoplanin for therapy of gram-positive infections in neutropenic patients. AB - Teicoplanin was evaluated in 20 febrile neutropenic patients as additional treatment for suspected Gram-positive infections after inadequate response to initial empiric ceftazidime monotherapy. Five patients with primary septicaemia, four with secondary septicaemia, 12 with localized infections and three with pyrexia of unknown origin were treated with teicoplanin (200 mg bolus intravenously, once daily after a 400 mg loading dose), whilst ceftazidime (2 g, 8-hourly, 30 min infusions) was continued. Four patients were unassessable (tuberculous, viral, protocol violation, and non-infectious pyrexial episode). Clinical cure for the combination was achieved in 11 or the 16 assessable cases (69%). Ten of the eleven (91%) bacteriologically confirmed infections were cured after addition of teicoplanin. Three strains of Staphylococcus aureus, four of Staphylococcus epidermidis (two methicillin resistant), and three strains of Streptococcus faecalis were isolated from successfully treated patients. One patient with Aerococcus and Enterobacter cloacae infection only improved after addition of erythromycin. One superinfection occurred with signs of interstitial pneumonitis in a patient following bone marrow transplantation. Neither ototoxicity nor nephrotoxicity occurred during treatment. A transient rise of liver transaminases was observed in four patients, but was attributable to teicoplanin in only one case. It is concluded that teicoplanin is a potentially effective and well-tolerated antimicrobial agent in neutropenic patients with infections due to Gram-positive organisms. PMID- 2964419 TI - [Between fear and pity: some socio-historical aspects of disability]. AB - Disability, like sickness, is not merely a physical condition, the state of an individual's body. It is also a collective status that relates the disabled person to Society. We analyze the evolution of this social status and we see how the perception of this phenomenon has changed over the ages. From this historical overview, the continuity as well as the discontinuity in changes can be observed, the changes that affect the institutions, values and conceptions having to do with disability. PMID- 2964420 TI - The right to prevention, early detection and medical rehabilitation under national and international aspects. AB - The importance of prevention and early detection and rehabilitation has been recognized and emphasized at national and international level for more than a decade. But systematic measures for implementation are still lagging far behind other rehabilitation activities. Civil rights legislation established the rights of citizens to protection of health, but health promoting and protecting measures must also be spelled out specifically in legislation. The most important aspects of such legislation are highlighted and illustrated by examples from the Federal Republic of Germany. The population needs information on possible prevention which should start in school. Human genetic counselling and prenatal counselling must be made readily available. Vigorous campaigns against substance abuse are vital. Pregnant women need special health protection support. Transportation, workplace and home must be made safer to name only a few areas in need of special legislation. Experiences in the beneficial effects of early detection have been made and documented in recent years and can serve as guidelines for further improvements. Assistance to developing countries in the areas of prevention, early detection and medical rehabilitation should become the most important task of development aid. PMID- 2964421 TI - Elderly handicapped persons in a primary care district: changes during a 5-8 year follow-up. AB - 643 handicapped persons born in 1918 or earlier were found at an inventory in 1975-78. In a follow-up 5-8 years later the mortality was 46%, which was significantly higher compared to the general mortality in the same age groups. There was no relationship between social class and mortality. Based on data from the first study, the deceased showed, compared to the surviving, impaired ADL and they used more medical and social care. The surviving population showed impairments in primary and secondary ADL during the follow-up. Elderly handicapped persons constitute a group at risk as they have an increased mortality and a great need of medical and social care. They should therefore be known to the medical profession. PMID- 2964422 TI - Common vocational training project for the handicapped (CVTPH). AB - CVTPH is a special project imparting vocational training to leprosy patients as well as other categories of the handicapped under the same roof, in order to combat the leprosy stigma which is the major obstacle in all leprosy work and to aid rehabilitation by making the trainees economically self-sufficient. The project, which offers training in the industrial sector to leprosy patients, orthopaedically handicapped individuals and able-bodied but socio-economically disadvantaged individuals, began in a small way in 1977, but has grown considerably since then, thus demonstrating the effectiveness of the concept. There have been several problems--chief among which are the leprosy stigma and the reluctance of the trainees themselves to move out from a sheltered environment into the world outside--but these are slowly being overcome by the formation of co-operative societies controlled by the handicapped. This project is hoped to serve as a model for similar vocational training programs. PMID- 2964423 TI - Metronidazole treatment in rosacea. PMID- 2964424 TI - BCG granulomas in a dysplastic nevus syndrome patient. PMID- 2964425 TI - Tyrosinemia type II with incomplete Richner-Hanhart's syndrome. PMID- 2964426 TI - Papillon-Lefevre syndrome. Successful treatment with oral retinoids in three patients. AB - Papillon-Lefevre syndrome is a rare autosomal recessive disorder of palmoplantar keratinization in which there is a unique association of the palmoplantar hyperkeratosis and premature loss of both deciduous and permanent teeth. We report three patients with papillon-lefevre syndrome who showed a remarkable degree of improvement after treatment with an oral retinoid. PMID- 2964427 TI - Serotonergic mechanisms in feeding. PMID- 2964428 TI - T cell subsets in children with mumps meningitis. AB - Nineteen children with mumps meningitis were investigated. Peripheral blood T cell subset function and number were tested during the acute phase of meningitis and again when the children recovered. No significant difference in the total number of T cells, or T helper: suppressor lymphocyte cell ratio was found in the children with mumps meningitis as compared with the controls. However, a significant increase in the number of T suppressor/cytotoxic cells in the peripheral blood of children with mumps meningitis in the acute phase was found. This increase in the T suppressor subset was still present even after the children recovered from the disease. Two children who showed deafness had the highest increases in the T suppressor/cytotoxic lymphocyte subset. Functional analysis of T suppressor cell activity demonstrated a significant suppressive activity of lymphocytes obtained during the acute phase of the disease. However, after recovery the suppressive activity found was not significant (P greater than or equal to 0.1). PMID- 2964429 TI - The inhibitory effect of sodium alginate on antibiotic activity against mucoid and non-mucoid strains of Pseudomonas aeruginosa. AB - The exopolysaccharide of mucoid strains of Pseudomonas aeruginosa (MPA) is chemically similar to alginate, a common polysaccharide of seaweeds. Both polymers have been reported to decrease the diameter of the inhibition zones produced in antibiotic disc diffusion assays. In order to determine whether this phenomenon was due to reduced diffusion of the antibiotic in an agar matrix, or to inhibition of antibiotic activity by sodium alginate, we studied the effect of sodium alginate on the susceptibility of P. aeruginosa strains to antibiotics in disc diffusion assays, as well as in broth and agar dilution assays. Sodium alginate decreased the activity of the aminoglycoside antibiotics, amikacin and gentamicin, against two MPA strains and their non-mucoid derivatives in each of the assays. In broth dilution assays, increase of the calcium ion concentration likewise reduced aminoglycoside activity against P. aeruginosa, but not against Escherichia coli and Staphylococcus aureus. Sodium alginate caused no inhibition of the activity of piperacillin and carbenicillin. The reduction of aminoglycoside activity may have implications for the common failure of these antibiotics in the treatment of pulmonary infections caused by MPA in cystic fibrosis patients, but must be considered only an in-vitro phenomenon at present. PMID- 2964431 TI - Problems in the assessment of outcome in a back pain clinic. AB - A sample of 72 patients attending the rheumatological back pain clinic of a district general hospital were interviewed at home, before their visit, about their expectations of the clinic. They also completed a Back Pain Disability Questionnaire. Fifty of the patients were recontacted by mail 3 months after their first clinic attendance and asked to complete the Back Pain Disability Questionnaire together with satisfaction questionnaires. Patients' reports, both with regard to expectations and satisfaction, particularly emphasized the importance of communication. One-third of the follow-up sample rated their clinic attendance as unhelpful, and although such views correlated with lack of improvement in pain and disability, other factors are involved in patients' evaluations. Significant correlations were obtained between subjective outcomes and health locus of control, social class and previous hospital treatment. Patient satisfaction may be a valuable measure of outcome in assessing the efficacy of back pain treatment. PMID- 2964432 TI - Driving, the law and medical disabilities. AB - There are 800 notifications daily to the Licensing Centre in Britain of medical disabilities. The procedure by which these are assessed is described, and the criteria for the issue of licenses to disabled drivers outlined. PMID- 2964430 TI - The effects of enoximone (MDL-17043) on forearm venous circulation in healthy volunteers and patients with heart failure. AB - The effect of intravenous enoximone on forearm venous circulation was studied in ten healthy volunteers (group A) and in ten patients with NYHA class III-IV congestive heart failure (group B). Distensibility of the forearm capacitance vessels was assessed from pressure-volume curves by venous occlusion plethysmography using a mercury-in-rubber strain gauge. Three recordings each at 3-min intervals were obtained before the infusion and again 20 min after completion of the infusion. Venous volume changes (VV) at congesting pressures of 10, 20, and 30 mmHg before and after enoximone were compared. Forearm muscle blood flow was also measured by venous occlusion plethysmography; electrocardiogram, heart rate, and cuff blood pressure were recorded throughout. Enoximone at a dose of 1 mg/kg body weight was infused over 10 min through a peripheral vein in group A and via a central line in group B. In group A, the effect of the injection vehicle was also assessed. VV10, VV20, and VV30 did not differ from baseline values after enoximone in both groups A and B. The vehicle caused a small but significant degree of venoconstriction in group A (VV20, 2.64 +/- 0.9 to 2.48 +/- 0.83 ml/100 ml, P less than 0.05; VV30, 3.47 +/- 1.27 to 3.33 +/- 1.20 ml/100 ml, P less than 0.05), which could be explained by an acute response to local pain from the infusion. This effect was not evident following enoximone, perhaps as a result of its counterbalancing vasodilating action to venoconstriction induced by acute pain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964433 TI - The assessment of people with arthritis who wish to drive a car. AB - In Great Britain 504,565 people are in receipt of Mobility Allowance; this, together with improved technology and an increase in public awareness, has increased the mobility aspirations of disabled people. Among those wishing to learn to drive for the first time, or to continue driving, are people with arthritis. In many cases pain and restriction of joint movement necessitate modification of car controls or driving technique. Between May 1982 and April 1985, 83 people with arthritis were assessed at Banstead. This paper describes the techniques used at Banstead Place Mobility Centre to assess these drivers, the difficulties which they experience, and the modifications which have been recommended. PMID- 2964434 TI - A 3-year follow-up of the driving status of 32 young adults with spina bifida. AB - Thirty-two young spina bifida adults with or without hydrocephalus were assessed for car driving potential in 1983. A postal questionnaire procedure was used to follow their decisions on driving and early driving experiences. This study reports the driving status (non-driver, learner driver or driver) of this group at December 1986 in relation to scores on the perpetual--cognitive battery completed on assessment. Results indicated that although nearly half of this group had passed the driving test, clinical assessment of cognitive functioning did not discriminate between drivers and non-drivers. PMID- 2964435 TI - Teaching the driving instructors. AB - Over the past 5 years 3-day courses for driving instructors have been held at the Banstead Place Mobility Centre. These courses are designed to familiarize instructors in the skills of teaching disabled people to drive. As a result 150 driving instructors throughout the British Isles are now on the National Register. PMID- 2964436 TI - A systematic approach to the management of the HIV positive individual. PMID- 2964437 TI - Re: Mandatory assignment--is it really legal? PMID- 2964438 TI - Bilateral distal femoral epiphyseal fractures following home delivery: a case report. PMID- 2964439 TI - The effects of non-steroidal anti-inflammatory agents on psoriasis. PMID- 2964440 TI - Angioscopic evaluation of vascular anastomoses of the lower extremities in the canine model. PMID- 2964441 TI - Adenocarcinoma of the prostate. PMID- 2964442 TI - Conformational changes in the vicinity of the N-iodoacetyl-N'-(5-sulfo-1 naphthyl)ethylenediamine attached to the specific thiol of sarcoplasmic reticulum Ca2+-ATPase throughout the catalytic cycle. AB - In the previous experiment (Suzuki, H., Obara, M., Kuwayama, H., and Kanazawa, T. (1987) J. Biol. Chem. 262, 15448-15456), the Ca2+-ATPase of sarcoplasmic reticulum vesicles was labeled with N-iodoacetyl-N'-(5-sulfo-1 naphthyl)ethylenediamine without a loss of the catalytic activity. The main labeled site was Cys674. A large monophasic fluorescence drop occurred upon ATP binding to the catalytic site of the Ca2+-activated enzyme in the presence of K+. The present results show that this fluorescence drop is biphasic in the absence of K+. The first and rapid phase of this drop accounts for most of the fluorescence drop. This phase reflects a conformational change in the enzyme.ATP complex. The second and slow phase, being much smaller than the first phase, coincides with phosphoenzyme (EP) isomerization from the ADP-sensitive form to the ADP-insensitive form. This phase disappears when accumulation of ADP insensitive EP is inhibited by K+ or when EP isomerization is prevented by the N ethylmaleimide treatment. These results show that this phase reflects a conformational change upon EP isomerization. When free Ca2+ is chelated after EP formation from ATP, the fluorescence intensity is restored to the initial level without Ca2+. This restoration coincides with EP decomposition. This suggests that the fluorescence restoration reflects a conformational change upon hydrolysis of ADP-insensitive EP. This probability is supported by the concurrent occurrence of the Pi-induced fluorescence drop and EP formation from Pi. The results demonstrate that the fluorescence drop upon ATP binding is predominant in the fluorescence change throughout the catalytic cycle. PMID- 2964443 TI - Regulation of ATP synthesis catalyzed by the calcium pump of sarcoplasmic reticulum. AB - The role of pH, KCl, ATP, water activity, and temperature in ATP synthesis from ADP and Pi was investigated in sarcoplasmic reticulum vesicles isolated from rabbit skeletal muscle. In totally aqueous medium, the synthesis of ATP was inhibited by ATP, KCl, and pH values above 6.5. When the water activity of the medium was decreased by the addition of 30% (v/v) dimethyl sulfoxide, the synthesis of ATP was no longer inhibited by ATP; it was activated by KCl and the optimum pH changed from 6.5 to 7.5. In totally aqueous medium, the concentration of MgCl2 needed for half-maximal synthesis of ATP was found to vary with the temperature of the assay medium; at 35 degrees C it was 1 mM and increased to a value higher than 10 mM when the temperature was decreased to 15 degrees C. In the presence of 30% dimethyl sulfoxide, maximal synthesis of ATP was attained in presence of 0.05 mM MgCl2 at both 15 and 35 degrees C. The hypothesis is raised that in the living cell water structure may play a role in regulating the synthesis of ATP observed during the reversal of the Ca2+ pump of the sarcoplasmic reticulum. PMID- 2964444 TI - Structural properties of a soluble bioactive precursor for transforming growth factor-alpha. AB - The precursor for transforming growth factor-alpha, proTGF-alpha, is synthesized as an integral membrane glycoprotein with the mature TGF-alpha sequence located in the extracellular domain. Retrovirally transformed rat embryo fibroblasts (FeSV-Fre cells) expressing the endogenous proTGF-alpha gene release and accumulate in the medium mature TGF-alpha as well as a heterogeneous (17-19 kDa) group of soluble, bioactive TGF-alpha precursor forms. These precursors correspond to the heterogeneously glycosylated extracellular domain of proTGF alpha which is released from the membrane by proteolytic cleavage. They are designated mesoTGF-alpha to denote their intermediate position in the proTGF alpha processing pathway. The nature of the carbohydrate linked to mesoTGF-alpha has been examined by treatment with glycosidases and the use of metabolic inhibitors of glycosylation. The results indicate that the TGF-alpha precursors from FeSV-Fre cells contain O-linked carbohydrate as well as sialylated N-linked carbohydrate. Heterogeneous N-linked glycosylation of an 11-kDa core polypeptide accounts for the heterogeneous nature of mesoTGF-alpha. MesoTGF-alpha released by cells treated with inhibitors of N-linked carbohydrate processing appears as a 17 kDa species. Treatment with these inhibitors does not alter significantly the production of mesoTGF-alpha or mature TGF-alpha by the cells. However, treatment of cells with an inhibitor of co-translational N-linked glycosylation, tunicamycin, reduces the accumulation of mesoTGF-alpha in the medium and blocks the production of mature TGF-alpha under conditions in which overall protein synthesis is only minimally affected. These findings suggest that the proTGF alpha processing activity is limiting in FeSV-Fre cells and other transformed cells that accumulate mesoTGF-alpha in the medium and that proTGF-alpha processing depends on a component whose function may require N-linked glycosylation. PMID- 2964445 TI - Purification and biological characterization of an adenovirus type 2 E1A protein expressed in Escherichia coli. AB - The adenovirus 2 E1A gene encodes a multifunctional protein of 289 amino acids that can immortalize primary rodent cells and transcriptionally activate a number of viral and cellular genes. To facilitate an understanding of the molecular basis for the various actions of E1A, we have redesigned our bacterial expression vector (Ko, J.-L., and Harter, M. L. (1984) Mol. Cell. Biol. 4, 1427-1439) containing the cloned E1A gene such that a soluble authentic E1A protein now constitutes approximately 1.5% of the total Escherichia coli cellular protein. Further, we have developed a simple rapid purification scheme without the use of detergents or denaturants and show a purity of greater than 98% with a yield of approximately 53%. The E1A so purified is biologically active, stimulating cellular DNA synthesis following microinjection into quiescent NIH 3T3 and REF52 cells. In another report (Spangler, R., Bruner, M., Dalie, B., and Harter, M. L. (1987) Science 237, 1044-1046) we have also shown that our purified E1A protein activates transcription from appropriate promoters in an in vitro system. PMID- 2964446 TI - Proteolytic processing of the alpha-chain of the lysosomal enzyme, beta hexosaminidase, in normal human fibroblasts. AB - The two subunits of beta-hexosaminidase undergo many post-translational modifications characteristic of lysosomal proteins, including limited proteolysis. To identify proteolytic cleavage sites in the alpha-chain, we have biosynthetically radiolabeled the transient forms, isolated these by immunoprecipitation, gel electrophoresis, and electroelution, and subjected them to automated Edman degradation. The position of the NH2-terminal amino acid was inferred from the elution cycle of the radioactive amino acid and the primary sequence encoded in the alpha-chain cDNA. The amino terminus of the precursor obtained by in vitro translation of SP6 alpha-chain mRNA in the presence of microsomes was leucine 23. The same amino terminus was found in precursor alpha chain synthesized by normal human fibroblasts (IMR90) in a 1- or 3-h pulse or secreted by these cells in the presence of NH4Cl. The alpha-chain isolated after a 3-h pulse followed by a 5-h chase (intermediate form) included a mixture of molecular species of which the amino terminus was arginine 87 (most abundant), histidine 88, or leucine 90. After a 20-h chase (mature form) the latter species predominated. This mature form of the alpha-chain remained fully reactive with antibody raised against the carboxyl-terminal 15 amino acids, indicating little if any proteolysis at the carboxyl terminus. Thus synthesis and maturation of the alpha-chain of beta-hexosaminidase includes two major proteolytic cleavages: the first, between alanine 22 and leucine 23, removes the signal peptide to generate the precursor form, whereas the second occurs between the dibasic amino acids, lysine 86 and arginine 87. The second cleavage is followed by trimming of 3 additional amino acids to give the mature form of the alpha-chain. PMID- 2964447 TI - Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G. AB - Chimeric Fc receptors, consisting of the IgG-binding domains of both staphylococcal protein A and streptococcal protein G, were constructed. An efficient bacterial expression system was used to produce the recombinant proteins, which vary in size and number of IgG-binding domains. The purified receptors were analyzed by immunodiffusion and a competitive enzyme-linked immunosorbent assay to establish the relative binding strength to various polyclonal and monoclonal immunoglobulins from different species. The results demonstrate that protein A and protein G have complementary binding patterns and that the chimeric receptors retain the binding capacities of both the parental constituents. This suggests that these novel chimeric receptors might be versatile reagents for immunochemical assays. PMID- 2964448 TI - Guanosine triphosphate promotes the post-translational integration of opsin into the endoplasmic reticulum membrane. AB - Membrane integration of a nascent opsin polypeptide was examined to determine whether insertion of proteins into the endoplasmic reticulum is dependent upon energy provided by ribonucleotide triphosphate hydrolysis. A discrete-sized nascent chain was obtained by in vitro translation of a mRNA which lacked a termination codon yet encoded the first 156 residues of bovine opsin. Ribosomes bearing the newly synthesized opsin chains were post-translationally incubated with canine pancreas microsomal membrane vesicles after addition of exogenous ribonucleotides or ribonucleotide analogues. Post-translational membrane integration and glycosylation of the 156-residue nascent polypeptide was found to require either the presence of guanosine triphosphate or a nonhydrolyzable GTP analogue. ATP did not promote post-translational integration of the nascent polypeptide. Although ribonucleotide hydrolysis was not obligatorily required for integration of opsin, we observed an increase in the proportion of glycosylated opsin chains in post-translational incubations that contained hydrolyzable ribonucleotide triphosphates. We conclude that a GTP-binding protein performs an essential role during integration of opsin into the endoplasmic reticulum. PMID- 2964449 TI - Provocation discography as a guide to planning operations on the spine. AB - In a prospective study we attempted to define the role of lumbar discography in the investigation of patients with low back pain with or without non-dermatomal pain in the lower limb. The records of 195 patients were studied at least two years after a technically successful operation. Of 137 patients in whom discography had revealed disc disease and provoked symptoms, 89% derived significant and sustained clinical benefit from operation. Of 25 patients whose discs showed morphological abnormality but had no provocation of symptoms on discography only 52% had clinical success. These findings support the continued use of lumbar discography for the investigation of this particular group of patients. PMID- 2964451 TI - Secretory vesicles externalize the major plasma membrane ATPase in yeast. AB - Yeast cell surface growth is accomplished by constitutive secretion and plasma membrane assembly, culminating in the fusion of vesicles with the bud membrane. Coordination of secretion and membrane assembly has been investigated by examining the biogenesis of plasma membrane ATPase (PM ATPase) in secretion defective (sec) strains of Saccharomyces cerevisiae. PM ATPase is synthesized as a approximately 106-kD polypeptide that is not detectably modified by asparagine linked glycosylation or proteolysis during transit to the plasma membrane. Export of the PM ATPase requires the secretory pathway. In sec1, a mutant defective in the last step of secretion, large amounts of Golgi-derived vesicles are accumulated. Biochemical characterization of this organelle has demonstrated that PM ATPase and the secretory enzyme, acid phosphatase, are transported in a single vesicle species. PMID- 2964450 TI - Intracellular movement of two mannose 6-phosphate receptors: return to the Golgi apparatus. AB - We have used Chinese hamster ovary (CHO) cells and a murine lymphoma cell line to study the recycling of the 215-kD and the 46-kD mannose 6-phosphate receptors to various regions of the Golgi to determine the site where the receptors first encounter newly synthesized lysosomal enzymes. For assessing return to the trans most Golgi compartments containing sialyltransferase (trans-cisternae and trans Golgi network), the oligosaccharides of receptor molecules on the cell surface were labeled with [3H]galactose at 4 degrees C. Upon warming to 37 degrees C, the [3H]galactose residues on both receptors were substituted with sialic acid with a t1/2 approximately 3 hrs. Other glycoproteins acquired sialic acid at least 8-10 times slower. Return of the receptors to the trans-Golgi cisternae containing galactosyltransferase could not be detected. Return to the cis/middle Golgi cisternae containing alpha-mannosidase I was measured by adding deoxymannojirimycin, a mannosidase I inhibitor, during the initial posttranslational passage of [3H]mannose-labeled glycoproteins through the Golgi, thereby preserving oligosaccharides which would be substrates for alpha mannosidase I. After removal of the inhibitor, return to the early Golgi with subsequent passage through the Golgi complex was measured by determining the conversion of the oligosaccharides from high mannose to complex-type units. This conversion was very slow for the receptors and other glycoproteins (t1/2 approximately 20 h). Exposure of the receptors and other glycoproteins to the dMM sensitive alpha-mannosidase without movement through the Golgi apparatus was determined by measuring the loss of mannose residues from these proteins. This loss was also slow. These results indicate that both Man-6-P receptors routinely return to the Golgi compartment which contains sialyltransferase and recycle through other regions of the Golgi region less frequently. We infer that the trans-Golgi network is the major site for lysosomal enzyme sorting in CHO and murine lymphoma cells. PMID- 2964452 TI - Mouse mammary epithelial cells produce basement membrane and cell surface heparan sulfate proteoglycans containing distinct core proteins. AB - Cultured mouse mammary (NMuMG) cells produce heparan sulfate-rich proteoglycans that are found at the cell surface, in the culture medium, and beneath the monolayer. The cell surface proteoglycan consists of a lipophilic membrane associated domain and an extracellular domain, or ectodomain, that contains both heparan and chondroitin sulfate chains. During culture, the cells release into the medium a soluble proteoglycan that is indistinguishable from the ectodomain released from the cells by trypsin treatment. This medium ectodomain was isolated, purified, and used as an antigen to prepare an affinity-purified serum antibody from rabbits. The antibody recognizes polypeptide determinants on the core protein of the ectodomain of the cell surface proteoglycan. The reactivity of this antibody was compared with that of a serum antibody (BM-1) directed against the low density basement membrane proteoglycan of the Englebarth-Holm Swarm tumor (Hassell, J. R., W. C. Leyshon, S. R. Ledbetter, B. Tyree, S. Suzuki, M. Kato, K. Kimata, and H. Kleinman. 1985. J. Biol. Chem. 250:8098-8105). The BM 1 antibody recognized a large, low density heparan sulfate-rich proteoglycan in the cells and in the basal extracellular materials beneath the monolayer where it accumulated in patchy deposits. The affinity-purified anti-ectodomain antibody recognized the cell surface proteoglycan on the cells, where it is seen on apical cell surfaces in subconfluent cultures and in fine filamentous arrays at the basal cell surface in confluent cultures, but detected no proteoglycan in the basal extracellular materials beneath the monolayer. The amino acid composition of the purified medium ectodomain was substantially different from that reported for the basement membrane proteoglycan. Thus, NMuMG cells produce at least two heparan sulfate-rich proteoglycans that contain distinct core proteins, a cell surface proteoglycan, and a basement membrane proteoglycan. In newborn mouse skin, these proteoglycans localize to distinct sites; the basement membrane proteoglycan is seen solely at the dermal-epidermal boundary and the cell surface proteoglycan is seen solely at the surfaces of keratinocytes in the basal, spinous, and granular cell layers. These results suggest that although heparan sulfate-rich proteoglycans may have similar glycosaminoglycan chains, they are sorted by the epithelial cells to different sites on the basis of differences in their core proteins. PMID- 2964454 TI - Suppression of postmenopausal ovarian steroidogenesis with the luteinizing hormone-releasing hormone agonist goserelin. AB - Twenty-one postmenopausal women with advanced breast cancer were treated with monthly 3.6-mg sc injections of the LHRH agonist goserelin [D-Ser-(But)6, Azgly10 LHRH] to determine whether the resultant endocrine changes could provide an explanation for the clinical responses that occur during therapy with this agent. After 4 weeks, serum gonadotropin levels were less than 10% of pretreatment levels, whereas serum PRL levels did not change. A significant decrease in serum testosterone occurred in 19 of 20 patients; this fall was associated with a 22% fall in serum estradiol levels. Serum androstenedione levels also decreased, but serum estrone and dehydroepiandrosterone sulfate (DHAS) levels did not. The lack of fall in serum DHAS levels indicates that the changes in androgen levels were a result of reduced ovarian secretion, and the reduced estradiol levels were a consequence of reduced precursor (i.e. testosterone) availability. The continued dependence of ovarian androgen secretion on gonadotropin stimulation after the menopause may explain the responses of some patients to LHRH agonists and some other therapeutic agents of unknown or uncertain modes of action. PMID- 2964455 TI - Studies of the effect of suppressor T lymphocytes on the induction of antithyroid microsomal antibody-secreting cells in autoimmune thyroid disease. AB - The suppressor function in CD8 (suppressor/cytotoxic) T lymphocytes from patients with autoimmune thyroid disease and normal subjects has been studied. CD8 and CD4 (helper/inducer) cells were separated by the panning method. Patient's non-T cells and autologous CD4 cells were cultured with or without autologous or allogeneic CD8 cells in the presence of either pokeweed mitogen or Staphylococcus aureus strain Cowan 1 plus human thyroid microsomal antigen. Antithyroid microsomal antibody (AMA) and total immunoglobulin G (IgG)-secreting non-T cells were measured by enzyme-linked immunosorbent spot assay. With pokeweed mitogen stimulation, the suppressor effect of CD8 cells from patients with serum AMA on the induction of AMA (of IgG type)-secreting cells was significantly less than that of CD8 cells from normal subjects. CD8 cells from patients with no serum AMA suppressed the induction of AMA-secreting cells as much as did normal CD8 cells. CD8 cells from both patients and normal subjects suppressed the induction of IgG secreting cells equally well. On the other hand, with the combination of S. aureus strain Cowan 1 and human thyroid microsomal antigen (1 mg/L) stimulation, CD8 cells from both normal subjects and patients only slightly suppressed the induction of IgG-secreting cells. However, under these circumstances, once again, CD8 cells from both normal subjects and patients with no serum AMA suppressed the induction of AMA-secreting cells, whereas CD8 cells from patients with serum AMA suppressed the induction of the AMA-secreting cells significantly less. Higher TMc concentrations enhanced the suppressor effect of CD8 cells from patients with serum AMA on the induction of AMA-secreting cells. Furthermore, Concanavalin A, when added to the stimuli described above, further inhibited the induction of both AMA- and IgG-secreting cells by CD8 cells from patients with serum AMA. There thus appears to be a relative defect of antigen-specific suppressor T lymphocyte function in CD8 cells from patients with autoimmune thyroid disease, which may result in the presence of autoantibody-secreting cells in those patients. PMID- 2964453 TI - Glomerular basement membrane proteoglycans are derived from a large precursor. AB - The basement membrane heparan sulfate proteoglycan produced by the Englebreth Holm-Swarm (EHS) tumor and by glomeruli were compared by immunological methods. Antibodies to the EHS proteoglycan immunoprecipitated a single precursor protein (Mr = 400,000) from [35S]methionine-pulsed glomeruli, the same size produced by EHS cells. These antibodies detected both heparan sulfate proteoglycans and glycoproteins in extracts of unlabeled glomeruli and glomerular basement membrane. The proteoglycans contained core proteins of varying size (Mr = 150,000 to 400,000) with a Mr = 250,000 species being predominant. The glycoproteins are fragments of the core protein which lack heparan sulfate side chains. Antibodies to glomerular basement membrane proteoglycan immunoprecipitated the precursor protein (Mr = 400,000) synthesized by EHS cells and also reacted with most of the proteolytic fragments of the EHS proteoglycan. This antibody did not, however, react with the P44 fragment, a peptide situated at one end of the EHS proteoglycan core protein. These data suggest that the glomerular basement membrane proteoglycan is synthesized from a large precursor protein which undergoes specific proteolytic processing. PMID- 2964456 TI - Distribution of D1 and D2 dopamine receptors in the basal ganglia of the cat determined by quantitative autoradiography. AB - The patterns of dopamine D1 and D2 receptors were examined in the corpus striatum and related structures in the cat brain by quantitative autoradiography after in vitro radioligand binding with [3H]SCH23390 (D1 antagonist) and [3H]spiperone (D2 antagonist). Highly specific binding for both radioligands occurs in striatal structures known to receive dopaminergic input: the caudate nucleus, putamen, nucleus accumbens, and olfactory tubercle. However, the density of binding varies from one structure to another, and the density distribution within striatal nuclei is heterogeneous. In all but one portion of the striatum, the concentration of bound D1 radioligand ranges from 46 to 230% more than that of the D2 radioligand. The exception to this difference occurs at caudal putamenal levels where the two radioligands bind in equal concentrations (approximately equal to 220 fmol/mg tissue wet-weight). The highest density of both D1 and D2 radioligand binding occurs in irregular zones in the head and body of the caudate nucleus. Such high-density zones of D2 radioligand binding appear mainly in the dorsolateral part of the caudate's head. For the D1 radioligand, the high-density zones are more widespread throughout the caudate nucleus, nucleus accumbens, and putamen. The D2 radioligand binding (but not the D1) also exhibits low-density zones at more caudal levels of the caudate nucleus, and these are often in register with the high-density zones of D1 radioligand binding. In the putamen, inverted concentration gradients exist for the two radioligands: the [3H]SCH23390 gradient runs from higher levels rostrally to lower levels caudally. The lowest levels of bound [3H]spiperone in the striatum occur in the nucleus accumbens olfactory tubercle area, whereas the lowest binding of [3H]SCH23390 occurs in the caudal putamen. Pallidal and nigral structures show marked disparities in binding of the two different radioligands. The D2 radioligand binding in the globus pallidus (80 +/- 8 fmol/mg tissue wet-weight) is about twice that in the entopedunuclear nucleus and pars reticulata of the substantia nigra, the latter two having equal levels (35 +/- 3 fmol/mg). No specific binding of the D2 radioligand occurs in the ventral pallidum. In contrast, D1 radioligand binding is highest in the entopeduncular nucleus (217 +/- 6 fmol/mg) and in the pars reticulata of the substantia nigra (198 +/- 2 fmol/mg) and moderate in the ventral pallidum (135 +/- 15 fmol/mg). In the globus pallidus, no detectable D1 radioligand binding occurs.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2964457 TI - Pruritus and the Leser-Trelat sign. PMID- 2964458 TI - Role of anxiety and anger in acne patients: a relationship with the severity of the disorder. AB - Studies investigating the possible role of personality and emotional factors in acne patients have yielded inconsistent and sometimes contradictory results. In the present study acne patients self-rated the severity of their conditions as mild, moderate, or severe and were compared to a control group on several personality variables. These self-ratings were compared to the objective ratings made by their dermatologists. Acne patients who self-rated their conditions as severe were found to experience significantly higher levels of trait anxiety than patients with mild and moderate conditions and control subjects; patients with severe and moderate conditions showed a higher state of anxiety than control subjects. Acne patients rated as severe by their dermatologists showed higher states of anxiety, "anger-in," and "anger-out" than the others. No other significant differences in terms of severity emerged, supporting the view that acne patients are not markedly neurotic. The results suggest that anxiety and anger are significant factors for severe acne patients and that future research should include severity ratings from both patients and physicians in the research design. PMID- 2964459 TI - Lymphomatoid papulosis and its relationship to "idiopathic" hypereosinophilic syndrome. AB - Persistent hypereosinophilia, endomyocardial fibrosis, and a recurrent self healing papulonodular eruption with the histologic features of lymphomatoid papulosis are described in three patients. One patient died after developing an acute myeloblastic transformation in the eosinophil series. Immunocytochemical studies of cutaneous lesions in two of the patients suggested a mature T-cell phenotype with a predominant population of CD4-positive cells. Immunostaining of cutaneous tissue with monoclonal antibodies BE1 and BE2 yielded negative findings. Because it is now known from in vitro studies that T lymphocytes secrete the eosinopoietic factor, interleukin 5, it is possible that the cutaneous lesions, hypereosinophilia, and associated endomyocardial fibrosis were induced by transformed helper T lymphocytes in these three patients. PMID- 2964460 TI - Bullous pemphigoid mimicking bullous erythema multiforme: an untoward side effect of penicillins. AB - Three young and middle-aged patients developed severe bullous eruption after receiving various penicillins. The clinical findings included high fever, prostration, widespread bullous eruption mainly on the head, face, and extremities, targetlike lesions on the palms and soles, and severe erosions in almost all the mucous membranes. Direct and indirect immunofluorescence studies were those of bullous pemphigoid. Complete clearing of the eruption occurred with prednisone therapy. We conclude that drug-induced bullous pemphigoid is a different entity from the classic bullous pemphigoid. PMID- 2964461 TI - Comparison of CO2 laser and electrosurgery in the treatment of rhinophyma. AB - A number of surgical modalities, including the CO2 laser, have been advocated for the treatment of rhinophyma. The CO2 laser has been extensively promoted for the management of many skin lesions, including warts, keloids, and various nail problems. In many instances it has been suggested that the CO2 laser holds a distinct advantage over other surgical modalities. Three patients with rhinophyma are presented. Results are compared after treating one half of the nose with biterminal electrocutting current and the other half with the beam of the CO2 laser. PMID- 2964463 TI - Acne rosacea in blacks--were Demodex mites found in any patients? PMID- 2964462 TI - Skin eruption caused by fat-soluble vitamin K injection. PMID- 2964464 TI - Agranulocytosis from isotretinoin. PMID- 2964465 TI - Long-term tetracycline use in the treatment of acne vulgaris--the role of routine laboratory monitoring. PMID- 2964466 TI - Measurement of nasal mucosal blood flow. PMID- 2964467 TI - A longitudinal study of use of health care services among older women. AB - This research tested the hypothesis that the percent of variance explained in use of health care services by the health care services utilization model could be significantly increased by including measures of past use of health care services and of past health status. Data from older women who participated in the Social Security Administration's Longitudinal Retirement History Survey (N = 1894) were analyzed by means of regression analysis. The results revealed that measures of previous use of health care services were more strongly related to current use of health care services in 1979 than were measures of previous health status. Inclusion of previous use and previous health status variables almost doubled the amount of variance explained by current predictors in number of physician visits, and more than doubled the explained variance in having to put off health care while the amount of variance explained in number of hospital episodes and in number of hospitalized nights was increased by approximately one-third. PMID- 2964468 TI - The therapeutic role of a structured life review process in homebound elderly subjects. AB - This study examined the therapeutic role of a structured life review process in a randomly selected group of 60 homebound elderly subjects. Subjects were placed in three groups and tested on four dependent variables at the beginning and end of an 8-week period. These variables were life satisfaction, psychological well being, depression, and activities of daily living (ADL). One group, the experimental group, received the treatment of life review process; another, the control group, received a friendly visit; and the third, the no-treatment group, received pretests and posttests only. Two dependent variables, life satisfaction, as operationalized by the Life Satisfaction Index A (LSIA), and psychological well-being, as operationalized by the Affect-Balance Scale (ABS), were significant in the experimental group when tested statistically through analysis of covariance (ANCOVA). These results suggest that a structured process of life review can serve as a therapeutic intervention for homebound elderly persons. PMID- 2964469 TI - Prevalence of disability in instrumental activities of daily living among elderly Japanese. AB - The prevalence of disability in instrumental activities of daily living (IADL) was assessed for seven items of activity -- among them, using public transportation, using the telephone, and shopping -- in 7,735 elderly residents living in an urban Japanese community. The prevalence of disability generally was low (ranging from 6.1% in heating water to 15.9% in preparing meals), but increased significantly with age. The prevalence was higher, controlling by age, in females than in males, with the exception of preparing meals. PMID- 2964470 TI - Modulation of lymphocyte subsets in Peyer's patches of mice treated with monoclonal antibody against helper T-cells. AB - Treatment of mice with anti-L3T4, a monoclonal antibody directed against helper T cells, impairs clearance of intestinal Giardia muris infection. The present study examined the effect of anti-L3T4 treatment on mouse Peyer's patch cytoarchitecture and on the distribution of T-cell subsets within microenvironments of the follicle. Female BALB/c mice, aged 8 weeks, were given 4 7 weekly injections of either anti-L3T4 (1 mg/wk) or PBS (control group), and Peyer's patches were examined by immunohistochemistry or flow cytometry. In anti L3T4-treated mice, Peyer's patch follicles (B-cell areas) were about two thirds the size of follicles in controls, and virtually all the size difference occurred in germinal centers. Peyer's patches were depleted of L3T4+ cells, yet the proportion of Thy-1.2+ (all T) cells was not decreased correspondingly, and the distribution of Thy-1.2+ cells in the patches was similar to that in control mice. In anti-L3T4-treated mice, Thy-1.2+ cells consisted of (a) Ly-2+ (cytotoxic/suppressor T) cells, and (b) a population of Thy-1.2+ cells that were neither L3T4+ nor Ly-2+. After treatment, Ly-2+ cells accounted for most of the T cells in interfollicular areas and were also scattered in follicles, in germinal centers, and below the dome epithelium--in areas where L3T4+ cells predominated in control mice. Thy-1.2+ cells that were L3T4- and Ly-2- were mainly localized below the dome epithelium. These shifts indicate complex interrelationships among different lymphocyte subsets in Peyer's patches. PMID- 2964471 TI - CD3/Ti gamma A: a functional gamma-receptor complex expressed on human peripheral lymphocytes. AB - We have recently developed a mAb designated anti-Ti gamma A, which was found to immunoprecipitate from the well characterized CD3+ TCR alpha/beta- F6C7 fetal clone a CD3-associated disulfide-linked gamma-glycoprotein. This antibody recognizes approximately 3% of adult peripheral lymphocytes and delineates a CD2+ CD3+ TCR alpha/beta- CD4- NKH1- subset where expression of CD8 appears to vary widely from one individual to another. In the present study, we have used anti-Ti gamma A mAb to assess whether gamma-chains expressed on these adult lymphocytes are used as functional R. The two activities which have been associated thus far with TCR gamma+ cells, that is, IL-2-dependent proliferation and non-MHC restricted cytotoxicity, were investigated here by using either resting or activated Ti gamma A+ lymphocytes. On the resting state, these cells (which appear as a very homogeneous population of granular lymphocytes) mediate little if any NK activity that could not be augmented by anti-Ti gamma A mAb. In contrast, after initial stimulation by PHA plus rIL-2 and subsequent culture in the presence of IL-2, activated Ti gamma A+ lymphocytes were strongly lytic against a series of conventional NK target cell lines. This cytotoxic function was either blocked or enhanced by anti-Ti gamma A mAb, depending upon experimental conditions. With respect to proliferation, it was possible to induce responses of resting Ti gamma A+ lymphocytes with antibody-coated CNBr beads only in the presence of exogenous IL-2, whereas, in culture, the same cells proliferated directly and secreted IL-2 after treatment by anti-Ti gamma A beads. Taken together, these data demonstrate that a major subset of circulating CD3+ TCR alpha/beta- lymphocytes use protein products of T cell gamma rearranging genes as functional R structures. PMID- 2964472 TI - Role of T cells in regulating expression of the B cell repertoire. Anti-ssDNA precursor frequency of DBA/2 B cells is increased in the presence of T cells from NZB mice. AB - Splenic B cells from DBA/2 and NZB mice were compared with regard to precursor frequency of anti-ssDNA-producing cells. Using a modification of the splenic fragment assay, we show that NZB T cells are capable of increasing the frequency of expression of anti-ssDNA precursors in DBA/2 splenic B cells. When limiting numbers of splenic B cells of DBA/2 origin were adoptively transferred into an irradiated (1200 rad) recipient, the co-transfer of NZB T cells markedly increased the frequency of anti-ssDNA precursors in cultured splenic fragments. The anti-ssDNA produced under these conditions was exclusively IgM and exhibited a high degree of cross-reactivity with TNP and fluorescein. Thus, the increase in anti-ssDNA precursor frequency reflected an expansion of the B cell repertoire to include precursors of polyspecific antibody-producing cells that under normal circumstances are not expressed. The ability of NZB T cells to increase the anti ssDNA precursor frequency was further defined by the CBA/N immunodeficiency gene xid, in that B cells from DBA/2.xid donors did not exhibit increased anti-ssDNA precursor frequency in the presence of NZB T cells. When NZB splenic B cells were co-transferred with DBA/2 T cells, the anti-DNA precursor frequency of the NZB B cells was not reduced. This study demonstrates that T cells can influence the emergency of B cell clones in an Ag-nonspecific manner. The well documented in vivo spontaneous polyclonal activation of NZB B cells may be secondary to T cell mediated expansion of the B cell repertoire. PMID- 2964473 TI - Isotype-specific immunoregulation. Systemic antigen induces splenic T contrasuppressor cells which support IgM and IgG subclass but not IgA responses. AB - In the present study, we have isolated and characterized the Lyt-1+, -2- T contrasuppressor (Tcs) cells from mice systemically primed with SRBC. Adoptive transfer of splenic Tcs cells from these mice abrogates oral tolerance and supports IgM and IgG anti-SRBC plaque-forming cell (PFC) responses; however, unlike the responses seen after transfer of Tcs cells derived from orally primed mice, low IgA responses were seen. Mice systemically primed with lower SRBC doses (0.01 to 1%) exhibited contrasuppression only within the L3T4- T cell subset, whereas mice primed with a high dose of SRBC (10%), harbored Lyt-1+, -2- Tcs cells in both the L3T4+ and L3T4- subsets. Both the L3T4- and L3T4+ Tcs cell subsets supported IgM and IgG responses when adoptively transferred to orally tolerized mice, and when added to tolerized spleen cell cultures. Splenic Tcs cells from systemically primed mice supported mainly IgG1 and IgG2b subclass anti SRBC PFC responses, a pattern also seen with Tcs cells derived from orally primed mice. Both L3T4+ and L3T4- Tcs cells from systemically primed mice exhibited well established characteristics of contrasuppressor cells including binding to Vicia villosa lectin and expression of I-J. The splenic effector Tcs cells which support IgM, IgG1 and IgG2b anti-SRBC PFC responses are antigen-specific, since both L3T4- and L3T4+ Tcs cells from spleens of mice primed with 10% SRBC reverse tolerance to SRBC, but not to horse erythrocytes (HRBC). Further, both L3T4- and L3T4+ Tcs cells from HRBC-primed mice reverse tolerance to IgM and IgG anti-HRBC, but not to anti-SRBC responses. Isolation of T3-positive Lyt-1+, -2- and L3T4- Tcs cell subsets by flow cytometry followed by adoptive transfer, showed that effector Tcs cells express T3 and presumably contain an Ag-R (TCR-T3 complex). These studies show that systemic priming with heterologous RBC induces splenic Ag specific Tcs cells in a dose-dependent manner, which support IgM and IgG subclass responses, but not IgA responses. PMID- 2964474 TI - Cytofluorometric analyses of human T cell CD2/CD4 inter-molecular interactions. AB - Incubation of human T lymphocytes with saturating concentrations of combinations of certain anti-CD2 and -CD4 mAb results in reciprocal down-regulation of the cell surface density expression of the respective CD molecules. Such reciprocal down-regulation occurs at 0 degrees C in the presence of sodium azide and appears selective for CD2 and CD4 molecules because mAb identifying various other CD T cell surface molecules (anti-Leu2a, -OK-CLL, -W6/32, -beta 2-microglobulin, -4B4) do not modulate CD2 or CD4 R density, and because anti-CD2 mAb (anti-OKT11 and D66 clone-1) do not alter CD8 R density (anti-OKT8, -Leu2a) and vice versa. Down regulation of CD2 by mAb specific to CD4 is epitope-specific but does not vary on the basis of the antibody isotype used. The anti-CD4 mAb, Leu3a, was the strongest CD2 down-regulator examined followed by OKT4F. mAb specific to other CD4 epitopes (B, C, D, and E) caused only slight down-regulation of CD2 expression whereas anti-OKT4 and -OKT4A mAb had no significant regulatory effect. Also, mAb specific to the 9.6 (anti-OKT11) and D66 (anti-D66 clone 1) epitopes of the CD2 molecule down-regulated CD4 density detectable with Leu3a, OKT4, and OKT4A anti-CD4 mAb. Down-regulation of CD2 by anti-CD4 mAb also occurred with the transformed T cell line, KE-37, which demonstrates that such effects can occur without mononuclear phagocytic accessory cells. From these data it can be concluded that important T cell immunoregulatory signals may be transmitted intramembranally between CD2 and CD4 glycoproteins. PMID- 2964475 TI - Intrathymic differentiation and tolerance induction of lymphokine-secreting Lyt 2+ T helper cells. AB - The present study has assessed thymic influence on the differentiation and recognition specificity of developing Lyt-2+ lymphokine-secreting T cells, and compared it with those of developing Lyt-2+ CTL. It was demonstrated that development of Lyt-2+ lymphokine-secreting Th cells requires an intrathymic differentiation step, and that peripheral Lyt-2+ lymphokine-secreting Th cells, unlike peripheral Lyt-2+ CTL, are profoundly tolerant to intrathymically expressed alloantigens. These data are interpreted as demonstrating that functionally distinct Lyt-2+ T cell populations are heterogeneous in their requirements for differentiation and/or activation. PMID- 2964476 TI - Loss of CD45R (Lp220) represents a post-thymic T cell differentiation event. AB - CD45R+ and CDw29+ CD4+ T cells are widely regarded as separate functionally defined T cell lineages. The work described here indicates that they represent maturation stages within the same differentiation pathway. Purified populations of CD4+ or CD8+ T cells, after stimulation with PHA, lose cell surface expression of CD45R (Lp220) and gain an increased surface density of CDw29 (4B4). Clonal analysis demonstrated that individual CD4+ CD45R+ T cells lost CD45R and acquired CDw29 with time in culture. This effect was selective for the high Mr 220-kDa form of the T200 (CD45) complex because the density of CD45, detected by an antibody to common determinants, did not decrease. This strongly indicates that CD45R+ cells are an immature stage in a lineage that culminates in CDw29 expression. To further define the expression of CD45R and CDw29, we analyzed infant thymus cells. Thymocytes include only 4 to 6% CD45R+ cells, but 95% express CDw29 in moderate density. The CD45R+ set appears to include mainly single CD4+ or CD8+, CD3 "bright" medullary cells, although only 15 to 25% of thymocytes with medullary phenotype express CD45R. In vitro culture of thymocytes with Con A and T cell growth factor induces expression of CD45R but these cells differ from the peripheral CD45R+ set by virtue of their co-expression of a high density of CDw29 (4B4) Ag. We postulate that post-thymically CD45R (Lp200) and CDw29 (4B4) comprise a functional assembly on the surface of T cells that changes in composition after stimulation with Ag or mitogen. This may result in enhanced ability of an Ag-experienced T cell to respond effectively to Ag due perhaps to a more efficient signaling complex. PMID- 2964477 TI - Decreased accessory cell function by human monocytic cells after infection with HIV. AB - We studied the effect of HIV infection on the human monocytic cell line U937. The cell line was infected with cellfree HIV, strain HTLV-IIIB. After 3 wk, a high reverse transcriptase activity was continuously detected in the supernatant of the cell line. Neither cytopathic effects nor changes in cell growth were observed. After infection, accessory cell function on T cell proliferation induced by anti-CD3 mAb of both IgG1 and IgG2a subclasses and Con A was tested. Accessory cell function provided by U937 cells started to decline 3 wk after inoculation with HIV. This correlated with detectable reverse transcriptase activity. The remaining accessory cell capacity varied between 10 and 60% of accessory cell function mediated by noninfected U937 cells. It was excluded that decreased FcR expression on U937/HIV cells contributed to the accessory cell defect in the anti-CD3-driven system. IL-2R expression on T cells, cocultivated with U937/HIV and anti-CD3, was minimal. The accessory cell defect could only be partly overcome by addition of rIL-2 or IL-1. Addition of high titer (10(4) TCID50) HIV or U937/HIV cells did not affect T cell proliferation, which rules out that the observed inhibition is caused by HIV infection of T cells or suppressive effects of U937/HIV cells. These results suggest that infection of APC may contribute to the induction of immunologic abnormalities in early HIV infection. Thus, monocytes/macrophages may not only serve as a reservoir for the dissemination of HIV, but may be an important target cell through which the immune system is affected. PMID- 2964478 TI - Expression of IgG Fc receptors in myeloid leukemic cell lines. Effect of colony stimulating factors and cytokines. AB - Three classes of FcR have been defined on human myeloid cells by their reactivity with mAb; FcRI (mAb 32); FcRII (mAb IV3); and FcRIII (mAb 3G8). We have quantitated the expression of each FcR on human myeloid leukemia cells and cell lines (KG-1, HL-60, U937, and K562). Detailed analysis of FcR surface expression is provided for the U937 cell line after exposure to CSF and cytokines. Increased expression of FcRI and FcRII occurred at 72 h in cells exposed to GCT or Mo cell line-conditioned medium as well as to medium from PHA-treated mononuclear cells. The augmentation of FcRII required protein synthesis and was diminished by a neutralizing antibody to granulocyte-macrophage CSF. We also show that fractions containing natural granulocyte CSF or granulocyte-macrophage CSF as well as r granulocyte and r-granulocyte-macrophage CSF are capable of inducing FcRII on these cells, whereas other cytokines such as IL-1 and IL-2, TNF-alpha, INF-gamma and macrophages CSF failed to do so. PMID- 2964479 TI - Autocrine growth of CD4+ T cells. Differential effects of IL-1 on helper and inflammatory T cells. AB - The growth factor requirements of cloned lines representing two major subsets of CD4+ T cells were examined. The helper subset, which produces IL-4 as its autocrine growth factor, proliferates in response to IL-2 or to IL-4 in the presence of IL-1. The inflammatory subset, which produces IL-2 as its autocrine growth factor, proliferates in response to IL-2 and, in the presence of limiting amounts of IL-2, shows increased proliferation in the presence of IL-4. The inflammatory subset does not proliferate in response to IL-1 plus IL-4. This ability to respond to the combination of IL-1 plus IL-4 correlates with the presence of IL-1R on the cloned lines tested. These data suggest that IL-1 may play a controlling role in the clonal expansion of CD4+ T cells of different functional types. This, in turn, suggests means by which the immune response could be directed into humoral or cell-mediated responses. PMID- 2964480 TI - Comodulation of CD3 and CD4. Evidence for a specific association between CD4 and approximately 5% of the CD3:T cell receptor complexes on helper T lymphocytes. AB - The aggregation of a specific class of lymphocyte surface molecules results in patching, capping, and surface modulation of the aggregated ligand. Both CD4, an associative recognition structure found on helper T lymphocytes, and CD3, a component of the T cell receptor complex, are members of this functional subgroup. When 125I-labeled monoclonal antibodies reactive with either CD4 (19Thy 5D7) or CD3 (RW24B6) were bound to T lymphocytes, the subsequent addition of goat anti-mouse Ig resulted in their rapid, temperature-dependent internalization. Whereas the binding of 125I-19Thy 5D7 (anti-CD4) was inhibited by greater than 90% in the presence of unlabeled 19Thy 5D7, no inhibition occurred in the presence of unlabeled antibody reactive with CD3 (RW28C8). We took advantage of the fact that these antibodies were of different isotypes (19Thy 5D7:IgG2a; RW28C8:IgGl) to determine whether the internalization of CD3 induced the comodulation of CD4. T lymphocytes preincubated with 125I-19Thy5D7 (anti-CD4) and unlabeled RA28C8 (anti-CD3) were treated with goat anti-mouse IgGl under conditions shown to quantitatively internalize CD3. After 1 h at 37 degrees C, T lymphocytes had internalized 10.5 +/- 2.6% (n = 3) of their antibody-bound cell surface CD4. After similar incubations with media alone or with goat anti-mouse IgGl in the absence of prebound RW28C8 (anti-CD3), no internalization of CD4 could be detected. Control antibodies reactive with CD45R (2H4, IgGl) also failed to induce the internalization of CD4. Similar results were obtained by using a helper T cell clone (T4C1) that internalized 9.6 +/- 2.8% (n = 3) of its antibody bound cell surface CD4 in response to CD3 modulation. In a reciprocal experiment, 125I-anti-CD3 (RW24B6, IgG2b) was preincubated with T4Cl cells together with unlabeled anti-CD4 (12T4D11, IgG1) prior to the addition of goat anti-mouse IgGl. The quantitative modulation of CD4 induced the co-internalization of 4.6 +/- 0.6% (n = 3) of cell surface CD3. These results suggest that approximately 5% of the CD3:T cell receptor complexes on helper T lymphocytes are specifically associated with CD4. Furthermore, our results suggest that an average of two CD4 molecules associate with each CD3:T cell receptor complex. PMID- 2964481 TI - Human epidermal cells from ultraviolet light-exposed skin preferentially activate autoreactive CD4+2H4+ suppressor-inducer lymphocytes and CD8+ suppressor/cytotoxic lymphocytes. AB - In vivo exposure of human epidermis to UV abrogates the function of T6+DR+ Langerhans cells and induces the appearance of Ag-presenting T6-DR+ OKM5+ cells in the epidermis. Since UV exposure of murine skin results in Ts lymphocyte activation, we investigated the capacity of human epidermal cells (EC) harvested 3 days after in vivo UV exposure to activate regulatory and effector autologous T lymphocyte subsets. T lymphocytes were separated into CD8+ suppressor/cytotoxic lymphocytes and CD4+ helper/inducer lymphocytes by C lysis and panning. The CD4+ subset was further divided by using the 2H4 mAB to obtain CD4+2H4+ lymphocytes (inducers of TS lymphocytes) and CD4+2H4- lymphocytes (inducers of B cell Ig production and inducers of cytotoxic T cells). Unirradiated suction blister derived EC from control skin (C-EC) and from skin exposed in vivo to UV (UV-EC) were cultured with purified autologous T lymphocyte subsets in the absence of added Ag. The resultant T lymphocyte proliferation was detected by [3H]thymidine uptake. UV-EC were highly effective in the stimulation of CD4+ lymphocytes, whereas C-EC were poor stimulators. The stimulator effect of UV-EC was abrogated after depletion of DR+ UV-EC. When CD4+ lymphocytes were fractionated, UV-EC consistently demonstrated enhanced ability to stimulate suppressor-inducer CD4+2H4+ lymphocytes relative to C-EC. Although less responsive than CD4+2H4+ lymphocytes, CD4+2H4- lymphocytes also demonstrated greater proliferation to UV EC than to C-EC. Neither UV-EC nor C-EC were able to activate CD8+ lymphocytes devoid of CD4+ lymphocytes. However, after addition of rIL-2 at concentrations that allow binding only to the high affinity IL-2R on T lymphocytes, UV-EC induced vigorous proliferation of CD8+ lymphocytes, whereas C-EC induced only background levels of proliferation. C lysis of leukocytes resident within UV-EC resulted in 66 to 70% reduction of CD8+ lymphocyte proliferation. In conclusion, UV-EC may activate CD8+ lymphocytes by at least two pathways: (1) UV-EC activation of CD4+2H4+ lymphocytes may induce differentiation/proliferation of CD8+ suppressor cells and (2) UV-EC activation of CD4+ cells may induce IL-2 production, that, in combination with UV-induced epidermal leukocytes, stimulates CD8+ cells. PMID- 2964482 TI - Differential regulatory signals delivered by antibody binding to the CD28 (Tp44) molecule during the activation of human T lymphocytes. AB - Molecule CD28 (Tp44) is expressed on the surface of majority of human T cells and has been implicated to play an active role in the regulation of T cell growth. The present study examines the effect of antibody binding to the CD28 molecule during T cell activation. Anti-CD28 but not isotype-matched anti-CD5 mAb consistently augmented anti-CD3-induced and IL-2-induced T cell proliferation and subsequent release of soluble CD25 molecule. When added together, mAb anti-CD28 and anti-CD5 acted synergistically to cause 2- to 7-fold enhancement of T cell activation induced by anti-CD3 mAb or IL-2 with no effect on the development of non-MHC-restricted IL-2-activated killer T cells. In contrast, alloantigen induced T cell proliferation, soluble CD25 release, and the subsequent development of CTL were all inhibited by anti-CD28 mAb. Moreover, alloantigen induced proliferative response of both CD4+ and CD8+ T cells was inhibited by anti-CD28 without affecting the cytolytic effect of CTL. Because valency of anti CD28 binding has been implicated as an important factor in signal transduction, this was explored in the allogeneic MLR by using Fab and F(ab')2 fragments of anti-CD28 mAb and anti-mouse kappa mAb. The inhibitory effect of anti-CD28 mAb in the MLR was reversed by cross-linking of anti-CD28 mAb with anti-mouse kappa mAb. In addition, cross-linking of the CD28 molecule on alloactivated T lymphoblasts but not that on resting T cells with anti-CD28 and anti-mouse kappa induced their proliferation in the absence of the priming alloantigen. These results indicate that stimulatory or inhibitory signals delivered through the CD28 molecule are determined by the degree of cross-linking of this molecule. In addition, these results also suggest that Ag-induced CD3-TCR-mediated T cell responses are more dependent on signals delivered through the CD28 molecule than those induced with anti-CD3, and thus these results have implications for potential use of anti-CD28 in sustained propagation of Ag-specific T cells. PMID- 2964483 TI - CD3-associated alpha/beta and gamma/delta heterodimeric receptors are expressed by distinct populations of CD4- CD8- thymocytes. AB - Immature double negative (DN) CD4-8- thymocytes expressing the CD3 molecular complex can be subdivided into two distinct subsets based on expression of the B2A2 antigen. Thus, B2A2+ CD3+ DN thymocytes express CD3-associated 35 kDa and 45 kDa polypeptide chains characteristic of a gamma/delta T cell receptor, whereas B2A2- DN thymocytes express predominantly 38 to 43 kDa CD3-associated structures typical of alpha/beta TCR. At the mRNA level, B2A2+ DN thymocytes express full length gamma and delta transcripts but no alpha message and only short (1 kb) B transcripts. In contrast, the B2A2- DN subset expresses full length alpha and beta transcripts and minimal levels of delta transcripts. Interestingly, B2A2- DN thymocytes, unlike mature T cells, also express abundant levels of full length TCR gamma mRNA, perhaps suggesting that these cells represent an early stage after divergence of the gamma/delta and alpha/beta T cell lineages. PMID- 2964484 TI - Proliferative and cytotoxic immune functions in aging mice. IV. Effects of suppressor cell populations from aged and young mice. AB - The changes with age in three splenic suppressor cell populations were studied in C57BL/6 mice. Allospecific Ts cells and nonspecific non-T suppressor cells were both generated in vitro in allogeneic MLC. The presence of "pre-existing" suppressor cells in fresh spleen cells from normal mice was examined. Suppressor cell activities were assayed for their ability to suppress proliferation in a fresh allogeneic MLC after treatment to prevent their own proliferation. The ability to generate both specific and nonspecific suppressor cells decreased with age, whereas pre-existing suppressor cells were detected in spleens from the majority of the aged animals but not in spleens from young animals. The decrease in suppressor cell activity was not due to any requirement for age matching between donors of suppressor and target cells. The specific and nonspecific MLC generated suppressor cells inhibited both the proliferative response in the assay MLC and the generation of cytotoxic cells. The pre-existing suppressor cells only suppressed the proliferative response and not the generation of cytotoxic cells. The changes seen with age in these suppressor cell populations suggest that the ability to generate suppression (both allospecific and nonspecific) to newly encountered Ag declines with age, whereas a resident splenic suppressor cell population accumulates over the lifetime of the animals. PMID- 2964485 TI - Sarcoidosis is not associated with a generalized defect in T cell suppressor function. AB - In pulmonary sarcoidosis, the marked expansion of CD4+ (helper/inducer) T cells in the alveolar structures of the lung is maintained by local IL-2 release by activated CD4+ HLA-DR+ T cells without concomitant expansion and activation of CD8+ (suppressor/cytotoxic) T cells, suggesting that sarcoid may be associated with a generalized abnormality of CD8+ T cells. Consistent with this concept, evaluation of the expression of the IL-2R on fresh lung T cells from individuals with active sarcoidosis demonstrated that 7 +/- 1% of sarcoid lung CD4+ T cells are spontaneously expressing the IL-2R compared with only 1 +/- 1% lung CD8+ T cells (p less than 0.01). However, stimulation of purified sarcoid blood CD8+ T cells with the anti-T3/TCR complex mAb OKT3 was followed by the normal expression of IL-2R (p greater than 0.1) and proliferation (p greater than 0.1). In addition, lung sarcoid CD8+ T cells responded to OKT3 similarly to normal lung CD8+ T cells and to autologous blood CD8+ T cells as regards expression of IL-2R (p greater than 0.1) and proliferation (p greater than 0.1). Finally, using CD4+ cells activated with allogenic Ag to induce, in coculture, fresh autologous CD8+ cells to suppress proliferation of fresh autologous CD4+ cells to the same Ag, sarcoid CD8+ T cells suppressed CD4+ cell proliferation in a normal fashion (p greater than 0.1). These results demonstrate that sarcoid CD8+ (suppressor/cytotoxic) T cells are competent to respond to a proliferation signal normally and can be induced to normally suppress CD4+ T cell proliferation to Ag, suggesting that the expansion of activated CD4+ T cells in pulmonary sarcoidosis is not due to a generalized abnormality of CD8+ T cells or of their suppressor T cell function. PMID- 2964486 TI - Flow cytometric analysis of human bone marrow. IV. Differential quantitative expression of T-200 common leukocyte antigen during normal hemopoiesis. AB - We have correlated the intensity of expression of CD45 Ag (T200 common leukocyte Ag) with mAb reactive with various lineages of hemopoietic cells in normal human bone marrow by using two-color immunofluorescence on a flow cytometer. Mature T lymphocytes (CD3+) and NK cells (CD16+ or CD11b+) expressed CD45 at the highest intensity. B lymphoid cells (CD19+) had three distinct levels of CD45 Ag expression. The bright CD45(3+) cells were mature B cells (CD19+, CD20+), whereas the less intense CD45(2+) cells were less mature B lymphoid cells (CD19+, CD10+). The dim CD45+ cells were very early, B lymphoid precursor cells (CD19+, CD10(2+), CD34+). The intensity of CD45 expression increased as cells matured in the monocytic lineage (CD14+, CD11b+). Among marrow granulocytic cells, CD45 intensity did not change on cells during maturation. Within the erythroid lineage, the most immature cells were CD45+ dim, and CD45 expression decreased during erythroid maturation to become undetectable on mature E. Hemopoietic progenitor cells (CD34+) expressed low levels of CD45 Ag. Expression of CD45R on marrow cells also showed intensity differences on different lineages. All NK cells (CD16+) were positive for CD45R, whereas only about one-half of the T lymphocytes (CD3+) were positive for CD45R. Almost all the cells in the erythroid and myelomonocytic lineages were CD45R-. Quantitative differences in expression of CD45R were observed on marrow B lymphoid cells which were correlated with the expression of CD45. The results show that quantitative changes in CD45 Ag expression accompany the differentiation and maturation of cells in the bone marrow. Comparisons with CD45R showed that this Ag was not always correlated with CD45. Since these Ag are the products of the same gene, these data indicate that the regulation of expression of the T200 molecules during normal hemopoietic development must be both quantitative and qualitative. PMID- 2964487 TI - Regulation of IgA secretion by T cell clones derived from the human gastrointestinal tract. AB - The role of T cells in Ig isotype regulation is still unclear. To address this question, we generated mitogen-stimulated T cell clones from normal human lymphoid follicles of the gut-associated lymphoid tissue (appendix). Both the T cell clones and clonal supernatants provided preferential help for IgA secretion by PWM-stimulated B cells. Many of these CD3+, CD4+, 4B4+, DR+ helper clones co expressed Fc-gamma and Fc-alpha R, but there was poor correlation between the expression of Fc-alpha R and IgA help (p = 0.31). Most of the T cell clones helped both IgM+A- and IgM-A+ B cell populations to secrete IgA, suggesting that they mediate switch of isotype-uncommitted B cells as well as post-switch expansion of IgA-committed B cells; however, some of the T cell clones helped IgM+A- B cell populations much more than IgM-A+ B cell populations, suggesting that, in this case, the regulatory effect is predominantly at the level of B cell switch. In all, these results show that the mucosal immune system contains individual T cells which are capable of positively regulating IgA-specific isotype differentiation at two levels of B cell development, thus allowing for efficient generation of IgA-secreting B cells. PMID- 2964488 TI - Regulation of the Ig kappa-chain enhancer by the adenovirus E1A gene products. Repression in lymphoid cells, activation in fibroblasts. AB - Proteins encoded by early region 1A (E1A) regulate transcription of viral and cellular genes. The mechanism of this trans-activation is not understood, but is of considerable interest as an example of transcription regulation through a cellular intermediate. We have therefore studied the effect of E1A products on the activity of the kappa L chain gene promoter and enhancer. By using transient and stable transfections into lymphoid and nonlymphoid cells, we found that the E1A proteins have a pleiotropic effect on the regulation of the mouse kappa-chain gene enhancer. In lymphoid cells the E1A products repress kappa-chain enhancer, whereas in fibroblasts, the kappa-chain enhancer is activated by the E1A products whether the E1A gene is in an extrachromosomal location or stably integrated in the genome. Furthermore, a functional kappa-chain promoter, containing the octanucleotide and "TATA" sequences is needed in order to be transcribed in E1A producing cells. This ability of E1A products to negatively and positively regulate kappa-chain transcription may reflect a more general phenomenon in which a given cellular protein could participate in a variety of different cellular controls. PMID- 2964489 TI - Use of OKT3 hybridoma cells to clonally activate CD3+ human T lymphocytes. AB - A simple and reliable method was developed to induce clonal growth of resting human T cells. In this limiting dilution (LD) culture system, responder cells (unseparated mononuclear cells, E rosette-purified T cells, or cell sorter separated CD4+ and CD8+ subsets) were activated by irradiated anti-CD3-secreting (OKT3) hybridoma cells in the presence of exogenous IL-2 (crude culture supernatant or recombinant IL-2). Under these conditions, one out of 2-3 CD4+ and CD8+ T cells developed into a proliferating cell clone. Addition of recombinant IL-1 slightly enhanced the growth frequency and increased the clone size of CD4+ cells but did not affect the growth pattern of CD8+ cells. PMID- 2964490 TI - Suppressor cell activation and enhanced skin allograft survival after tumor promotor but not initiator induced depletion of cutaneous Langerhans cells. AB - During chemical carcinogenesis Langerhans cells (LC) are depleted from the epidermis, disrupting the normal immunological functions of the skin. Tumor promotors but not initiators, have been shown to deplete adenosine triphosphatase (ATPase)-positive LC from the skin and therefore the cutaneous immune system may be impaired during tumor promotion but not initiation. The present study shows that the tumor promotor 12-O-tetradecanoylphorbol 13-acetate (TPA) but not the initiator urethane depletes Ia-positive LC from BALB/c murine ear epidermis, and beta-glucuronidase-positive LC from C57BL mouse tail skin. Sensitization with 2,4 dinitrofluorobenzene (DNFB) through urethane-treated skin resulted in a normal contact sensitivity response when the mice were challenged 5 days later. In contrast, tolerance resulted from sensitization through TPA-treated skin as a result of the generation of suppressor cells. In addition, TPA but not urethane treated C57BL mouse tail skin survived for an extended time when grafted onto histoincompatible BALB/c mice. Therefore, impairment of the normal immunological functions of skin resulted from treatment with the tumor promotor TPA but not the tumor initiator urethane, which suggests that a loss of LC during tumor promotion may impair immunological protection against skin tumors. PMID- 2964492 TI - Fatty acids of acylceramides from comedones and from the skin surface of acne patients and control subjects. AB - Comedonal lipids and skin surface lipids were collected from six acne patients and surface lipids were collected from sex- and age-matched controls without acne. Six series of ceramides were found in each sample, the relative amounts of which were determined by thin-layer chromatography/photodensitometry. Acylceramides (ceramide 1) were isolated by preparative thin-layer chromatography and their ester-linked fatty acids were analyzed by gas-liquid chromatography. The comedonal acylceramides contained higher proportions of 16:0, 16:1 delta 6, and 18:1 delta 6 + delta 8 and much less linoleate (18:2 delta 9,12) than the acylceramides from the skin surface. In the surface lipids from legs, acylceramides from the acne patients contained less linoleate than the acylceramides from control subjects. Free fatty acids from the comedones were also isolated and analyzed, and had a composition very similar to the esterified fatty acids of comedonal acylceramides. The results confirm that fatty acids derived from sebum become incorporated into comedonal acylceramides, displacing linoleate, and show that this process even affects the acylceramides of surface epidermis, more so in acne patients than in normal subjects. PMID- 2964491 TI - Parallel recovery of epidermal antigen-presenting cell activity and contact hypersensitivity responses in mice exposed to ultraviolet irradiation: the role of a prostaglandin-dependent mechanism. AB - Contact hypersensitivity (CH) responsiveness to 2-4-dinitro-1-fluorobenzene (DNFB) is depressed in mice that are sensitized through skin sites exposed to ultraviolet radiation (UVR). This is partially due to a reduction in antigen presenting cell (APC) activity within UVR-exposed skin, a condition marked by a decrease in the density of ATPase/Ia-positive epidermal cells. The purpose of this study was to correlate the histological and functional recovery of APC activity in the skin of C3H mice exposed to low-dose (4 X 450 J/m2) or high-dose (1 X 15 kJ/m2) UVR with the normalization of CH responsiveness. Skin biopsy specimens taken at various intervals after UVR exposure revealed a rapid recovery in the density of ATPase/Ia positive cells: about 70% of normal by 3 days, and normal after 5 days. Functional analyses showed that lymph node cells obtained from donors that were sensitized with DNFB 3 days after UVR treatment transferred normal ear-swelling responsiveness to non-primed recipients, thus indicating that APC activity in UVR-exposed skin paralleled the recovery of ATPase/Ia-positive epidermal cells. This suggested that an alternative mechanism causes the persistent depression of CH in mice exposed to UVR. Mice pretreated with indomethacin prior to UVR exposure demonstrated a capacity to elicit CH responses to DNFB, which paralleled the histological and functional recovery of APC in the skin (i.e., normal CH responses were elicited 3 days after exposure to UVR). We conclude from this study that APC activity in the skin recovers rapidly after exposure to UVR, and that a PG-dependent mechanism is responsible for many of the persistent and systemic effects that cause a depression in the CH responsiveness of mice treated with UVR. PMID- 2964493 TI - Sparing of leprosy macule in ampicillin hypersensitivity rash. AB - A case of tuberculoid leprosy is reported in a middle aged female, who presented with generalised erythematous maculopapular rash which followed ampicillin therapy. In spite of extensive and severe involvement of the skin all over the body by erythema and rash, the hypopigmented patch of leprosy on the face did not show any erythema or rash and stood out more clearly as an island of pale area in the midst of large area of erythema on the face. The possible mechanism of non development of erythema and rash in the patch is discussed. PMID- 2964494 TI - [Peroperative angiography in reparative surgery of the lower limbs. Apropos of experience with 1099 cases]. AB - The authors report their experience of per-operatory angiography in a series of 1,099 cases over 13 consecutive years. The main interest is the limitation in the rate of early reintervention, which frequently causes morbidity and mortality in this type or surgery. The serial nature of the technique is emphasized, and is most comparable to pre-operatory exploration. The various technical details are specified. Some hemodynamic troubles noted during injection (aortography) contraindicate the method in high-risk subjects. The main indications are, above all, control of restorative acts, but also a per-operatory diagnosis of a lesion, and the assessment of blood stream at a lower level. The results are morphological and the various anomalies discovered are reviewed, with iconography, in relation to each type of surgical act (disobliteration, bypass). In situ bypasses are considered separately. The data are also hemodynamic, and justify the six successive X-ray film technique. Deblocking (embolectomy with a balloon catheter) and endarterectomies were found to give more technical imperfections than bypasses. The rate of extemporaneous corrections tripled during the second period (1978 to 1986), and this was no doubt due to an extension in operative indications (19% versus 6% at the beginning). Dacron bypasses (fibrinous debris) and in situ bypasses (detection of anatomical abnormalities, location of shunts) were more often incriminated in immediate reinterventions than Dardik homografts or PTFE. PMID- 2964495 TI - [Reliability and perspectives in peroperative flowmetry. An original computer assisted ultrasonic technic]. AB - To mitigate the lack of reliability of currently used flowmetry technics, an original method was developed based on a single window 15 MHz Doppler flowmeter and an Apple IIe computer with data acquisition system. Eleven successive measurements of instantaneous rate were made in an arterial section. A mean flow was determined by integration of mean rate in section. The reliability of the method was evaluated by an experimental study on a hydrodynamic bench reproducing physiologic flow conditions with an error always less than 8%. An in vivo study in 30 patients provided 100 measurements of arterial flow before and after surgical arterial reconstruction. The mean error value was less than 10%. Among the risks of error, those due to the probe was reduced by specially conceived probes. The most important factor was that of respiration, this latter factor imposing an integration time of rate of signal of 30 seconds to eliminate errors due to respiration. The conventional measurement time for other configurations is 6 seconds but this leads to a mean error of 25% which can attain 60%. The perspective of miniaturization of the apparatus in the near future with acceleration of measurements using a multi-window Doppler should allow flow measurement in 30 seconds and control of surgical reconstructions (on healthy arteries, vein grafts and prostheses but excluding PTFE). Similarly, it should be possible to improve evaluation of so-called vasoactive drugs. PMID- 2964496 TI - CD16. Developmentally regulated IgG Fc receptors on cultured human monocytes. AB - We have demonstrated that one Fc receptor for IgG (FcR) (CD16) on cultured human monocytes appears to be a developmentally regulated membrane protein. This receptor appears to contain less carbohydrate (if any) than does its counterpart on human neutrophils. Expression of CD16 on cultured monocytes increases with respect to both percentage of positive cells and numbers of sites per cell with length of time in culture. This was in contrast to expression of other types of FcRs that either decreased (CDw32) or did not change (FcRp72). Unlike an FcR that binds monomeric IgG (FcRp72), expression of CD16 on monocytes from most normal individuals was not influenced by IFN-gamma. After 14 d in culture, CD16 appeared to be the predominant FcR on cultured monocytes, and was capable of mediating both ligand attachment and phagocytosis. These findings support the hypothesis that CD16 plays an important role in mediating immunophagocytosis. PMID- 2964497 TI - Role of T cell subsets in lethal graft-versus-host disease (GVHD) directed to class I versus class II H-2 differences. I. L3T4+ cells can either augment or retard GVHD elicited by Lyt-2+ cells in class I different hosts. AB - Detailed information was sought on the capacity of purified Lyt-2+ cells to mediate lethal graft-versus-host disease (GVHD) directed to class I H-2 differences. When B6 Lyt-2+ cells were transferred to irradiated class I different (B6 x bm 1)F1 mice, three different patterns of lethal GVHD were observed. First, rapid death from hematopoietic failure occurred when Lyt-2+ cells were transferred together with host-type marrow cells; this form of GVHD probably reflected direct destruction of stem cells by Lyt-2+ cytotoxic cells. Second, a pattern of late-onset, chronic GVHD resulting in death only after 4-6 wk occurred when Lyt-2+ cells were supplemented with donor marrow. This syndrome developed in the apparent absence of L3T4+ cells and was observed with either high or low doses of Lyt-2+ cells and with either light or heavy irradiation of the host. Third, an acute form of GVHD resulted when Lyt-2+ cells plus donor marrow cells were supplemented with exogenous help, i.e., by adding small doses of donor L3T4+ cells or injecting the hosts with rIL-2. Although L3T4+ cells potentiated GVHD when injected in small doses, supplementing Lyt-2+ cells with large doses of L3T4+ cells paradoxically led to marked protection; symptoms of GVHD were mild and no deaths occurred. PMID- 2964498 TI - Effects of beta-lipotropin, beta-endorphin, gamma 2-melanotropin and corticotropin on steroid production by isolated human adrenocortical cells. AB - Normal human adrenocortical cells (from multi-organ donors) were incubated with corticotropin (tetracosactide), highly purified human beta-lipotropin, synthetic human beta-lipotropin, gamma 2-melanocyte stimulating hormone and beta-endorphin. Corticotropin stimulated cortisol, aldosterone and 18-hydroxycorticosterone production starting at 10(-12)-10(-11) mol/l in normal adrenocortical cells. Purified human beta-lipotropin also stimulated steroidogenesis but 100-1000-fold higher concentrations of the peptide were needed. In contrast, synthetic human beta-lipotropin was without any effect on steroid production up to concentrations of 10(-7) mol/l. Synthetic beta-lipotropin (5 x 10(-10) mol/l) did not significantly change the dose-response curve for corticotropin (10(-13) mol/l-10( 9) mol/l) versus the three steroids measured. gamma 2-Melanotropin and beta endorphin (10(-6) mol/l) stimulated the secretion of cortisol, but not of aldosterone. Since synthetic human beta-lipotropin has no effect on human adrenocortical cells, the purified beta-lipotropin must be contaminated with traces of corticotropin. Since pathologically elevated levels of proopiomelanocortin-derived peptides will rarely exceed plasma concentrations of 10(-10) mol/l, our findings in vitro do not support a physiological or pathophysiological role of the peptides examined in the regulation of adrenal steroid secretion. PMID- 2964499 TI - An enzyme immunoassay for phospholamban. AB - Phospholamban, a phosphorylatable protein of the cardiac sarcoplasmic reticulum, has been estimated by a semi-quantitative immunoassay. It can be determined in purified membrane preparations as well as in crude fractions of cardiac muscle membranes, regardless of the phosphorylation state of the phosphoprotein. The content of phospholamban in mammalian heart muscle membrane vesicles correlates with the activity of the calcium/magnesium-dependent ATPase with the exception of the oxalate-loaded membrane preparations. This observation indicates that phospholamban and the calcium transporting enzyme are localized at different sites in cardiac sarcoplasmic reticulum. PMID- 2964500 TI - Practice effects in backward masking. AB - In two experiments we demonstrate that much larger practice effects occur in a backward masking paradigm where patterned masks are used than in similar visual processing paradigms, such as lateral masking and whole report. In additional experiments we examine four possible explanations for the large practice effects: increased familiarity with the paradigm in general, learning about the targets, learning about the masks, and enhanced sensory processing. Because of failure to observe similar practice effects in related paradigms not involving backward masking and because of the sustained nature of the improvement, we reject the first explanation as a source of practice effect. Experiment 3 allowed us to reject target learning as a source of improvement as well; target sets were switched at the end of training, but no decrement in performance was observed. In Experiment 4, mask sets were switched at the end of training, revealing a significant decrement in performance. Learning about the specific masks, then, does contribute to the observed improvement. However, it is responsible for only about one third of the overall improvement in performance. The final experiment provides evidence that the residual improvement is due to enhanced sensory processing. In that experiment, training on backward masking led to a lowered threshold in a two-flash paradigm but not to a significant change in whole-report performance. PMID- 2964501 TI - Probability and category redefinition in the fault tree paradigm. AB - Fault trees have been advocated as aids for problem solvers. However, research has suggested limitations in their usefulness. Fischhoff, Slovic, and Lichtenstein (1978) found that subjects given incomplete (pruned) trees were insensitive to omissions; these authors hypothesized that Tversky and Kahneman's (1974) availability heuristic was the mediating factor. Using a within-subjects design, subjects in Experiment 1 received both full and pruned trees and estimated probabilities for various reasons why a car would fail to start. To increase the availability of omissions, some Experiment 1 subjects first generated possible causes of starting failure. The basic Fischhoff et al. findings were replicated, but several aspects of the results argued against the availability hypothesis as the mechanism for judgment. Instead, subjects appeared to idiosyncratically redefine category membership when making judgments based upon pruned trees. By employing a sorting task in Experiment 2 we corroborated the results of Experiment 1: Subjects do redefine the actual contents of the categories when faced with an omission from the fault tree. The implications of these results for the use of fault trees as a problem solving aid are discussed. PMID- 2964502 TI - Effect of irrelevant differences as a function of the relations between relevant and irrelevant dimensions in the same-different task. AB - The effects of irrelevant differences as a function of the relations between relevant and irrelevant dimensions in the same-different task were examined. Form, size, and orientation were used as task conditions in Experiment 1, and form, size, and color were used in Experiment 2. In each experiment, 6 subjects were instructed to report same or different according to a relevant dimension, irrespective of two irrelevant dimensions. In Experiment 3, the degree of integrality was examined in all the combinations of dimensions involved, in the restricted-classification task. The results of the three experiments suggested that (a) effects of irrelevant differences depended on the degree of integrality between relevant and irrelevant dimensions, and (b) two irrelevant dimensions were processed by the subjects serially. Neither the relevance rechecking model (Miller & Bauer, 1981) nor the response competition model (e.g., Williams, 1974) alone could explain all the types of effects of irrelevant dimensions obtained in this study. Instead, a modified relevance rechecking model, in which the degree of integrality was introduced to the original relevance rechecking model, could predict and explain all types of effects. PMID- 2964503 TI - Convergent validation of information processing constructs with Pavlovian methodology. AB - These two experiments introduce a new nonverbal measure of stimulus structure by merging Garner's (1974) distinction of separable and integral stimulus properties with the field of Pavlovian conditioning. According to one rule, two sets of stimulus (separable vs. integral) were constructed. In each experiment a differential compound conditioning procedure was used with one group of subjects who were trained with a subset of the separable material and then were tested with an unfamiliar subset. The same procedure was used with a second group but with integral materials. Both experiments showed a reversal of conditioning effects, depending on the properties of the stimuli. Separable compounds showed results as predicted by elementaristic conditioning theories: The associative values of the elements summed up. With integral compounds, the associative values of the elements were irrelevant. Instead, integral compounds were processed and classified primarily on the basis of similarity. PMID- 2964504 TI - Mental rotation: effects of dimensionality of objects and type of task. AB - The original studies of mental rotation estimated rates of imagining rotations that were much slower when two simultaneously portrayed three-dimensional shapes were to be compared (R. Shepard & J. Metzler) than when one two-dimensional shape was to be compared with a previously learned two-dimensional shape (Cooper and her associates). In a 2 X 2 design, we orthogonally varied dimensionality of objects and type of task. Both factors affected reaction times. Type of task was the primary determiner of estimated rate of mental rotation, which was about three times higher for the single-stimulus task. Dimensionality primarily affected an additive component of all reaction times, suggesting that more initial encoding is required for three-dimensional shapes. In the absence of a satisfactory way of controlling stimulus complexity, the results are at least consistent with the proposal that once three-dimensional objects have been encoded, their rotation can be imagined as rapidly as the rotation of two dimensional shapes. PMID- 2964505 TI - Force variability in isometric responses. AB - In the present study we examined the contribution of different impulse parameters to peak force variability in an isometric task. Five experiments are reported that each held constant a different impulse parameter while allowing the other impulse parameters to vary. The results indicate that change in force level is the parameter that has the greatest effect on peak force variability, although time to peak force and preload also systematically influence response variability. A formula that accommodates the relation between impulse parameters and force variability is proposed. The data suggest that even in isometric tasks, it is the force-time properties of the impulse, rather than discrete parameters such as peak force, that determine the outcome variability. PMID- 2964506 TI - Localization of tactile stimuli and body parts in space: two dissociated perceptual experiences revealed by a lack of constancy in the presence of position sense and motor activity. AB - Two motor acts were analyzed at the level of tongue and fingers. These motor acts generated illusions. When subjects voluntarily rotated the tongue by 90 degrees, the perceived orientation of a tactile stimulus applied to the tongue did not covary with the perceived orientation of the tongue itself. Analogously, when subjects voluntarily crossed two adjacent fingers, the perceived position of two tactile stimuli applied to the fingers did not covary with the perceived position of the fingers themselves. Although tongue and fingers were positioned accurately in space, a lack of perceptual constancy occurred for tactile stimuli applied to these body parts. Therefore, whereas position sense was preserved, correct localization of objects was lost. The occurrence of this perceptual dissociation suggests that spatial localization of tactile stimuli may be independent both of knowledge of body part location and motor activity. PMID- 2964507 TI - Progression-regression effects in tracking repeated patterns. AB - Subjects used a position control system to perform compensatory tracking of a repeated input pattern. Tracking error was roughly proportional to the velocity of the input signal. Error magnitude decreased with practice and increased with the addition of a concurrent memory task. These effects can be modeled as progressive and regressive changes in how well subjects used control movement velocity and displayed error velocity to anticipate the input pattern and thereby reduce their effective time delay. The weighting of velocity cues in this model progressed with practice and regressed with the secondary task, even though the secondary task required no concurrent visual scanning or simultaneous motor response. This regression effect appears to indicate cognitive interference with the anticipation process. Stationary linear models provide a good approximation to the movement patterns; however, these models do not account for episodes of rapid pulse-like movements that were revealed in the ensemble-averaged trajectories. PMID- 2964508 TI - Global precedence in attended and nonattended objects. AB - In four experiments we evaluated whether there is a priority in early perceptual analysis for the global aspect of a nonattended, visual, compound stimulus. In all experiments, two objects were distinguished by being enclosed in either a square or a circle, and each object consisted of a larger letter constructed from smaller letters. Attention was directed to one of the two objects on the basis of either the square/circle distinction (Experiments 1, 2, and 3) or a visual precue (Experiment 4). In agreement with previous findings, for attended objects, the global aspect was identified faster and was more difficult to ignore than the local aspect. However, for nonattended objects, no evidence was found for an overall priority in perceptual processing leading to identification of the global aspect. Rather, the results indicate that there is a priority in perceptual processing for the general category of the global information contained within nonattended objects. Although the results provide some support for the view that global information receives priority during early perceptual analysis, aspects of the results are more consistent with postidentification accounts of global precedence. PMID- 2964509 TI - Acetylcholinesterase in Huntington's and Alzheimer's diseases: simultaneous enzyme assay and immunoassay of multiple brain regions. AB - A newly developed enzyme-linked immunosorbent assay for acetylcholinesterase (AChE) protein was combined with conventional measures of enzyme activity in a study of 15 brain regions from six control cases (non-neurological deaths), six cases of Alzheimer's disease, and six cases of Huntington's disease. In the control brains, the mean AChE activity varied 100-fold from region to region (cortex lowest, striatum highest). The variation in enzyme activity was exactly paralleled by a variation in protein immunoreactivity. Overall, the homospecific activity of AChE averaged 0.26 +/- 0.007 mU/pg, close to the value for electrophoretically homogeneous enzyme isolated from red blood cells. Similar homospecific activities were observed in samples from Huntington's and Alzheimer's brains. Evidently, AChE that is immunoreactive but enzymatically inactive does not accumulate in any of the three conditions examined. Huntington's brain samples showed normal total contents of AChE, but Alzheimer's brains showed significant decreases of both enzyme activity and immunoreactivity in all seven cortical regions and in two out of the eight subcortical structures examined, hippocampus and nucleus accumbens. PMID- 2964510 TI - Uptake of gamma-aminobutyric acid by a synaptic vesicle fraction isolated from rat brain. AB - Gamma-Aminobutyric acid (GABA) was taken up by a MgATP-dependent mechanism into synaptic vesicles isolated by hypoosmotic shock and density gradient centrifugation. The properties of the vesicular uptake differed clearly from those of synaptosomal and glial uptake, both with respect to Na+, Mg2+, and ATP dependence and with respect to response to general GABA uptake inhibitors such as nipecotic acid, diaminobutyric acid, and beta-alanine. The uptake showed a Km of 5.6 mM and a net uptake rate of 1,500 pmol/min/mg of protein. It is suggested that the vesicular uptake of GABA is driven by an electrochemical proton gradient generated by a Mg2+-ATPase. PMID- 2964511 TI - Monoclonal antibodies with high affinity for spiroperidol. AB - A diverse panel of monoclonal antibodies was obtained from BALB/c mice immunized with two haptens structurally related to spiroperidol (SPD). Bromoacetyl derivatives of aminospiroperidol (NH2SPD) and N-amino-phenethylspiroperidol (NAPS) were synthesized to couple the haptens covalently to a protein carrier for immunization, thereby maintaining the butyrophenone portion of the immunogen. Hybridomas were selected based on their ability to secrete antibody that binds [3H]SPD with high affinity. Equilibrium dissociation constants for these antibodies ranged from 0.2 to greater than 100 nM. The antigen binding sites of the anti-NH2SPD and anti-NAPS antibodies were characterized in studies of the inhibition of the binding of [3H]-SPD by a series of ligands that are either (a) structurally related to SPD or (b) structurally unrelated to the butyrophenones but known to be selective antagonists of the D2 subtype of dopamine receptor. Based on the patterns of inhibition of the binding of [3H]SPD by these compounds, 12 classes of antibody combining sites were identified. Most of these antibodies bound butyrophenones with high affinity. One anti-NH2SPD and four anti-NAPS antibodies also bound domperidone, a nonbutyrophenone that has a high affinity for D2 receptors. None of the antibodies bound clebopride or sulpiride, D2 selective antagonists of the benzamide class, or the agonist dopamine. PMID- 2964512 TI - Hereditary progressive chorea without dementia. AB - A family with hereditary non-Huntington's chorea is presented. Transmission was autosomal dominant with variable penetrance. Chorea commenced in childhood and affected predominantly the head, face and upper limbs. Dysarthria appeared later, followed in two family members by elements of an axial dystonia. There was no intellectual impairment. Unlike previously described families, symptoms progressed steadily up to the eighth decade, causing considerable physical disability. PMID- 2964513 TI - Topography of nerve cell loss from the locus caeruleus in middle aged persons with Down's syndrome. AB - A topographical analysis of nerve cell loss from the locus caeruleus in middle aged patients with Down's syndrome (whose brains show the pathological changes of Alzheimer's disease), has shown that cell loss is confined to dorsal areas, being least most rostrally and greatest caudally. By contrast, there is no significant cell loss from ventral parts of the locus, at any point along its rostrocaudal length. Dorsally located neurones of the locus project to cerebral cortex; ventrally located neurones to non-cortical areas such as basal ganglia, cerebellum and spinal cord. These data suggest that the damage to nerve cells of the locus caeruleus in Down's syndrome at middle age, like that seen in Alzheimer's disease itself, relates to primary pathological events within the cortical projection fields of affected cells with perikaryal loss following on as a later change. PMID- 2964514 TI - Histochemical characteristics of human mimic muscles. AB - Eight different mimic muscles of 13 human cadavers (7 male, 6 female) were studied by histochemical techniques. In sections stained for myosin ATPase the composition of fibre types was quantitatively evaluated by computer-assisted image analysis. The data were compared to those of 2 muscles in the lower limb of the same individuals. According to the percentage of the type I muscle fibres 3 groups of mimic muscles were distinguished: (1) the orbicularis oculi muscle (15%), (2) the major zygomatic, levator labii superioris, levator anguli oris, depressor anguli oris muscles and platysma (27-38%), and (3) the occipitofrontal and buccinator muscles (57-77%). For comparison, the gracilis and rectus femoris muscles were built up by 36% and 48% of type I fibres. The average diameters of fibres in mimic muscles were significantly less than in the 2 limb muscles. Differences in muscle fibre size between male and female specimens were not significant. The relevance of morphological characteristics of mimic muscles for facial expression and reconstructive surgery is discussed. PMID- 2964515 TI - Growth inhibitory effect of recombinant alpha and beta interferon on human glioma cells. AB - Growth inhibitory activity of recombinant alpha and beta interferon on two human glioma cell lines, EFC-2 and KE cells, was determined by two different growth assays. Recombinant beta interferon showed a slight growth inhibitory effect on EFC-2 cells at day 3, and maximum inhibition was seen on day 6 with an ID50 of 50 U/ml. Recombinant alpha interferon showed no significant growth inhibition at any concentration. KE cells were resistant to both recombinant alpha and beta interferon. The growth inhibitory activity of recombinant beta interferon on EFC 2 cells was not blocked by recombinant alpha interferon, although recombinant alpha and beta interferons shared same receptors on EFC-2 cells. Addition of DFMO (alpha-difluoromethylornithine) to interferon in the media showed additive effect rather than synergistic effect in growth inhibition of glioma cells. Out of 7 glioma cell lines tested, 4 showed heterogeneous sensitivity to recombinant beta interferon, and all were resistant to recombinant alpha interferon. These results suggest a differential sensitivity of EFC-2 cells to recombinant beta interferon, as well as a heterogeneous sensitivity to recombinant beta interferon among different glioma cell lines. PMID- 2964516 TI - Technetium-99m MAG3, a comparison with iodine-123 and iodine-131 orthoiodohippurate, in patients with renal disorders. AB - A Phase I study in 12 patients with renal disorders compared the simultaneous clearances of 99mTc-labeled mercaptoacetyltriglycine (MAG3) and 131I-labeled orthoiodohippurate (OIH). The ratio of MAG3 to OIH clearance was 0.61 +/- 0.08 as a result of its smaller volume of distribution, ratio 0.65 +/- 0.09, for the clearance half-lives were similar, ratio 1.09 +/- 0.12. A Phase II study performed serially in 20 patients with equal doses of [99mTc]MAG3 and [123I]OIH gave images of equal quality. The relative renal functions were highly correlated (r = 0.97, p less than 0.001) and transit time analyses gave good correlations: parenchymal transit time index r = 0.81, p less than 0.05. We conclude that [99mTc]MAG3 has some advantages over [99mTc]DTPA and is a suitable replacement for [123I]hippuran in routine renal imaging, relative function, and transit time studies, but not for the accurate estimation of the renal plasma flow. PMID- 2964517 TI - Immunogenicity of hepatitis B vaccine in children with sickle cell anemia. PMID- 2964518 TI - Duration of ductal shunting in healthy preterm infants: an echocardiographic color flow Doppler study. AB - The purpose of this investigation was to assess the duration of ductal shunting after birth in healthy preterm infants (30 to 37 weeks gestational age) without evidence of respiratory distress. Thirty-six infants were evaluated in the first 12 hours of life by means of two-dimensional echocardiography and color flow Doppler techniques, and then once daily until no ductal flow was detected (defined as functional closure). Preterm infants were subdivided into two groups by gestational age: group 1 = 30 to 33 weeks (n = 12); group 2 = 34 to 37 weeks (n = 24). Sixteen full-term infants (38 to 41 weeks) were similarly evaluated as control subjects (Group 3). One infant from each group had a closed ductus at the time of the first study (performed at a mean of 7.7 +/- 3.2 hours). Subsequent studies for the entire group were performed at a mean of 31.3 +/- 5.4 hours (day 2), 55.0 +/- 4.5 hours (day 3), and 80.3 +/- 6.1 hours (day 4). For the three groups, the rates of ductal closure ranged from 50.0% to 58.3% on day 2 and 81.3% to 87.5% on day 3. For the entire group, all but one infant had demonstrated closure of the ductus arteriosus by day 4. Within the range of gestational ages studied, we conclude that prematurity, in the absence of respiratory distress syndrome, does not prolong the initial duration of physiologic ductal shunting. PMID- 2964519 TI - The relative merits of various methods of indirect measurement of intraabdominal pressure as a guide to closure of abdominal wall defects. AB - Visceral ischemia secondary to increased intraabdominal pressure (IAP) following closure of abdominal wall defects presents a serious postoperative problem. Currently, the method of closure and postoperative management are determined by clinical impressions rather than measurement of IAP. In this study various methods of indirectly measuring IAP were compared in 17 rabbits in which IAP was sequentially increased with an intraabdominal balloon. Vesical and inferior vena caval (IVC) pressures were found to have good statistical correlation with IAP. Other methods tested were gastric, rectal, superior vena caval, femoral and brachial artery, and rectus compartment pressures. All were found to be poor indicators of actual IAP. In nine of the rabbits, radiolabeled microspheres were used to assess cardiac output and visceral blood flow. Renal blood flow was very sensitive to increased IAP with dramatic impairment at IAP above 10 to 15 mmHg. Small intestinal flow was less sensitive and did not become significantly diminished until IAP exceeded 25 to 30 mmHg. Our studies suggest that vesical and IVC pressure monitoring should be used to evaluate IAP in the clinical setting. If IAP is in excess of 10 to 15 mmHg surgical intervention is indicated to prevent the development of renal ischemia. PMID- 2964520 TI - Periodontal surgery for a patient with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an uncommon blood stem cell disorder. The acquired defect is in the cell membrane of the precursor of the affected clone, which renders erythrocytes, platelets and granulocytes unusually sensitive to the hemolytic effects of complement. It is characterized by chronic hemolysis, intermittent hemoglobinuria, thrombotic events and bone marrow hypoplasia. Hemolysis is precipitated by infection, strenuous exercise, surgery or menstruation. The dilemma of managing a PNH patient is that both the periodontal infection and the surgical treatment are precipitating factors. Management is further complicated by the fact that the patient is receiving corticosteroids. The treatment of the PNH patient is complex and potentially hazardous. It requires close cooperation between the periodontist and the hematologist. This paper describes the management of one such case. PMID- 2964521 TI - Sex hormone effects on personality at puberty. AB - In this study we use serum steroid hormone assays and Adjective Check List responses from a representative sample of 102 male and 99 female adolescents to examine the relations between hormones, personality, and sexual behavior. We address two questions: (a) Does pubertal testosterone (T) cause sex dimorphism in personality? (b) Do pubertal hormones affect sexual behavior indirectly through effects on personality? Exploratory factor analysis of the Adjective Check List generated a factor common to male and female adolescents that correlated highly with T. However, male and female subjects did not differ in their mean level on this factor or in the correlation of this factor with T. In spite of the large sex difference in T, the girls were much more sensitive than the boys on the extracted factor to differences in T. We conclude that the answer to both questions is probably no. PMID- 2964522 TI - Differential suppressor effects of the ssb-1 and ssb-113 alleles on uvrD mutator of Escherichia coli in DNA repair and mutagenesis. AB - We have constructed double mutants carrying either ssb-1 or ssb-113 alleles, which encode temperature-sensitive single strand DNA binding proteins (SSB), and the uvrD::Tn5 allele causing deficiency in DNA helicase II, and have examined sensitivity to ultraviolet light (UV), recombination and spontaneous as well as UV-induced mutagenesis. We have found in a recA+ background that (i) none of the ssb uvrD double mutants was more sensitive to UV than either single mutant; (ii) the ssb-1 allele partially suppressed the strong UV sensitivity of uvrD::Tn5 mutants; (iii) in the recA730 background with constitutive SOS expression, the ssb-1 and ssb-113 alleles suppressed the strong UV-sensitivity caused by the uvrD::Tn5 mutation; (iv) in ssb-113 mutants, the level of recombination was reduced only 10-fold but 100-fold in ssb-1 mutants, showing that there was no correlation between the DNA repair deficiency and the recombination deficiency; (v) the hyper-recombination phenotype of the uvrD::Tn5 mutant was suppressed by the addition of either the ssb-1 or the ssb-113 allele; (vi) no addition of the spontaneous mutator effects promoted by the uvrD::Tn5 and the ssb-113 alleles was observed. These results suggest a possible functional interaction between SSB and Helicase II in DNA repair and mutagenesis. PMID- 2964523 TI - Kinetics and mechanisms of etodolac degradation in aqueous solutions. AB - The extent and mechanisms of etodolac (1; 1,8-diethyl-1,3,4,9 tetrahydropyrano[3,4-b]-indole-1-acetic acid) decomposition, as a function of pH and temperature, were investigated. Three main degradation products derived from 1 are identified as 7-ethyl-2-(1-methylenepropyl)-1H-indole-3-ethanol (2), the decarboxylated product of etodolac (3), and 7-ethyltryptophol (4). The main pathway for the degradation of 1 is followed by consecutive first-order kinetics: 1----2 in equilibrium 3----4. No appreciable buffer effect on the degradation of 1 is observed for any of the buffer species in the study. The rate-pH profile exhibits a specific acid catalysis at acidic (kH) and neutral (k'H) pH regions, and an inflection point at pH 4.65 corresponding to the pKa value. From Arrhenius plots, the activation energies (Ea) for kH and k'H were found to be 26 and 24 kcal/mol, respectively. The small positive entropy of activation (delta S) indicates that a unimolecular decomposition mechanism is favored for the decomposition reaction of 1. PMID- 2964524 TI - 125I-Bolton-Hunter-8-methoxy-2-[N-propyl-N-propylamino]tetralin as a new selective radioligand of 5-HT1A sites in the rat brain. In vitro binding and autoradiographic studies. AB - In vitro binding assays with 125I-[8-methoxy-2-[N-propyl-N-(3'-iodo-4' hydroxyphenyl)-propionamido -N'- propylamino] tetralin] (125I-BH-8-MeO-N-PAT), a 125I-labeled derivative of the potent serotonin (5-HT) agonist 8-hydroxy-2-[di-n propylamino]tetralin [( 3H]-8-OH-DPAT), showed that this compound recognized specific sites with nanomolar affinity for 5-HT and 5-HT1A ligands such as spiroxatrine, ipsapirone, buspirone and gepirone in rat hippocampal membranes. Comparison of the binding characteristics of 125I-BH-8-MeO-N-PAT with those of [3H]-8-OH-DPAT revealed striking similarities: at the hippocampal level, both binding sites exhibited nanomolar affinity for their respective ligands and the same Bmax; their pharmacological profiles defined by the inhibition of each bound ligand by a series of 26 serotonin, dopamine- or norepinephrine-related agonists and antagonists were identical; and their regional distributions examined by membrane binding assays and autoradiography of labeled brain sections were highly correlated. These observations indicate that 125I-BH-8-MeO-N-PAT is the first 125I-reversible ligand for the selective labeling of 5-HT1A sites in the rat central nervous system. PMID- 2964525 TI - A quantitative study of finishing and polishing techniques for a composite. PMID- 2964526 TI - Late reconstruction of the patellar ligament using a Dacron ligament implant. PMID- 2964527 TI - Are the medical needs of mentally handicapped adults being met? AB - This paper reports on the apparent inadequate level of primary medical care provided to many of the 151 mentally handicapped people who attend an adult training centre. A large number of common medical problems were identified that were not known to the general practitioners and/or were not being managed, including problems known to be associated with Down's syndrome. Many trainees were further handicapped by unmanaged defects of hearing and vision. Contact rates with the general practitioner showed that the mentally handicapped adults did not place a greater burden on the doctor than the rest of the population. Comparing these rates with those for other vulnerable groups such as those aged over 75 years and under four years showed that only 28% of the trainees had an adequate consultation rate with the general practitioner.A lack of awareness among general practitioners of the special needs of this group is thought to be in part responsible but the major factor is the inherent problem of communication which exists almost universally in people who suffer from mental handicap. Ways of improving the situation are discussed with an emphasis on the need for a change in our attitudes towards mentally handicapped people. Reference is made to the desirability of increasing the cooperation between primary care and community mental handicap teams and the increasing importance of voluntary organizations. PMID- 2964528 TI - Possible evidence for genetic predisposition to nondisjunction in man. PMID- 2964529 TI - Beta endorphin, a vasoconstrictor during septic shock. AB - A relationship between increased peripheral resistance (TPRI) and decreased cardiac index (CI) and mortality from sepsis has been suggested. The relationship between endogenous opiates and this response was evaluated. METHODS: Chronically instrumented sheep were given E. coli endotoxin (LPS, 1.5 mcg/kg x 30 minutes). In one study, survivors (n = 9) and nonsurvivors (n = 11) of LPS were compared along with survivors (n = 8) of half the dose of LPS. In a second study, two groups of animals received naloxone: one (n = 11) had a bolus of 2 mg/kg followed by a 2 mg/kg/hr continuous infusion started 30 minutes before LPS while the other had the bolus and infusion started 1 hour after LPS was begun. RESULTS: Both vasoconstrictive and vasodilative phases were seen. Vasoconstriction was associated with elevated beta endorphin levels, a pattern sustained until death in the nonsurvivors. Both pre- and posttreatment with naloxone lessened the maximum increase in total peripheral resistance index compared with untreated sheep. DISCUSSION: The vasoconstrictive aspects of the response to LPS correlated with elevated beta endorphin levels and with mortality. This vascular response is attenuated with naloxone blockade. PMID- 2964530 TI - Standardization indices of cardiac hypertrophy in weight lifters. AB - In order to investigate further the use of standardization procedures to determine training effects on cardiac dimension and function, two groups of subjects were analysed noninvasively. A control group of sedentary men (n = 7) and an experimental group of weight lifters (n = 12) volunteered for a standard M mode echocardiographic assessment. Indices of cardiac function as well as absolute left ventricular dimensions and left ventricular mass were similar between the groups. Standardizing for body surface area and body weight separated the groups. The weight lifters were shown to have a more muscular left ventricular posterior wall. The weight lifters also demonstrated a dilated left ventricle when indexing for body surface area. All other dimensional and volumetric indices were non-significant. The present investigation underlines the need for matching control and experimental groups to biometric variables in order to avoid misinterpreting cardiac enlargement. There is evidence for a true relative hypertrophy in weight lifters as indicated by similar absolute cardiac dimensions and similar biometric variables. Therefore, when evaluating athletes engaged in a chronic pressure overload, cardiac hypertrophy indices should consider body surface area (BSA), body weight and lean body weight (LBW). Future work in this field should incorporate rigorous controls on all biometric variables for better interpretation of hypertrophy in relative or absolute terms. PMID- 2964531 TI - Nucleotide sequence of the tail sheath gene of bacteriophage T4 and amino acid sequence of its product. AB - The nucleotide sequence of gene 18 of bacteriophage T4 was determined by the Maxam-Gilbert method, partially aided by the dideoxy method. To confirm the deduced amino acid sequence of the tail sheath protein (gp18) that is encoded by gene 18, gp18 was extensively digested by trypsin or lysyl endopeptidase and subjected to reverse-phase high-performance liquid chromatography. Approximately 40 peptides, which cover 88% of the primary structure, were fractionated, the amino acid compositions were determined, and the corresponding sequences in DNA were identified. Furthermore, the amino acid sequences of 10 of the 40 peptides were determined by a gas phase protein sequencer, including N- and C-terminal sequences. Thus, the complete amino acid sequence of gp18, which consists of 658 amino acids with a molecular weight of 71,160, was determined. PMID- 2964532 TI - The use of deep duplex scanning to predict hemodynamically significant aortoiliac stenoses. AB - Hemodynamic assessment of aortoiliac occlusive disease is necessary for successful arterial reconstruction of the aorta and legs. Various methods have been proposed and "pull-through" intra-arterial pressures are the "gold standard." Deep Doppler duplex imaging was supplemented with real-time spectral analysis and velocity measurements in 29 cases. Twenty-three of these patients needed arteriography. One hundred sixty-six (166) arterial segments extending from the proximal aorta to the common femoral arteries were independently graded on duplex scans and arteriograms. For severe occlusive disease, duplex scanning is highly accurate (sensitivity 82%, specificity 93%). Velocity measurements were useful in determining the hemodynamic significance of stenoses. Peak systolic velocities in stenoses were measured with a duplex scanner. The pressure gradient calculated with the modified Bernoulli equation (delta P = 4Vmax2) correlated well with the gradients measured during arteriography (r = 0.9, n = 11). These noninvasive velocity measurements and Bernoulli calculations alert arteriographers to obtain special views of suspected areas and suggest the need for "pull-through" pressures and possible balloon angioplasty. In addition, these noninvasive measurements are useful to follow up patients who have mild to moderate aortoiliac disease and after angioplasty. PMID- 2964533 TI - Salvage of in situ femoropopliteal and femorotibial saphenous vein bypass with interventional radiology. AB - Arteriovenous fistulas and late graft stenoses are well-known potential causes of in situ saphenous vein bypass failure. Three patients are described who had one or both of these complications postoperatively (early and late). Two techniques are described that can be performed at the time of arteriography to save a functioning bypass. PMID- 2964534 TI - Buttock claudication from isolated bilateral internal iliac arterial stenoses. AB - An unusual case is reported of severe buttock claudication in a woman with normal ankle systolic pressures after exercise, for which the cause was eventually found to be isolated bilateral hypogastric arterial stenosis. Although a normal ankle pressure response to exercise usually rules out vascular obstruction in patients with symptoms suggestive of intermittent claudication, the diagnosis of isolated hypogastric arterial disease should be entertained when a neurogenic or orthopedic explanation can be excluded. PMID- 2964535 TI - [Two cases of IBL-like T-cell lymphoma showing helper/inducer characteristics]. AB - Two cases of an IBL-like T-cell lymphoma are reported, in one case of which the histological changes could be observed throughout its course. Although both cases were difficult to differentiate from true IBL in their early stage, focal proliferation of pale cells appeared in their first specimens and gradually, the characteristics of lymphoma grew manifest. Immunohistochemical studies of both cases revealed lymphoma cells having the characteristics of helper/inducer T lymphocytes. Electron-Microscopic examination showed no particular differences in the morphology of the tumor cells between our case and the case previously reported as being an IBL-like T-cell lymphoma of a suppressor/cytotoxic character. PMID- 2964536 TI - [Experimental study on splanchnic circulation by ultrasonic transit time volume flow meter and laser Doppler flow meter]. PMID- 2964537 TI - [Prevalences of allergic diseases among synthetic chemical workers]. AB - We studied the prevalence of allergic diseases among synthetic chemical workers, using a cross-sectional questionnaire, to clarify the epidemiological evidence of occupational allergy due to chemicals which workers were handling and manufacturing. A file, registered in 1981, of 6,819 person, was used to calculate the prevalence during the preceding 12 months of "allergy," "asthma," and "skin disease," and to analyze the relationship between the prevalence and the exposure risks such as the work or the chemicals being manufactured. The main results were: 1) In workers who were exposed to the various chemicals, the prevalence of "allergy," "asthma," and "skin disease" were 3.2, 1.9, and 9.0%, respectively. 2) Among the workers who were engaged in the manufacturing of synthetic resins, paints, films, titania or pharmaceuticals, the prevalence of "allergy" was significantly higher than among others. The rates of "asthma" were significantly higher than others among workers in cosmetics, synthetic resins, or fertilizers. The rates of "skin disease" were significantly higher among those working with cosmetics, synthetic resins, paints, dyes, or organic solvents. The results indicate that allergy research should be concentrated on workers engaged in manufacturing the above chemicals. PMID- 2964538 TI - Urinary screening for disorders of heteroglycan metabolism. Results of 10 years experience with a comprehensive system. AB - The results of 10 years experience in urinary screening for disorders of heteroglycan metabolism are presented. Over 5,000 urines were analysed of which 216 were positive for excess mucopolysacchariduria. The enzymatic diagnosis was achieved in 159 mucopolysaccharidoses of which Type III Sanfilippo was the commonest (86 cases), followed by Type II Hunter (31 cases) and Type I Hurler (21 cases). A total of 27 urines were positive for excess oligosacchariduria, the enzymatic diagnosis being established in 20 cases. The most frequently encountered oligosaccharidosis was GM1 gangliosidosis (10 cases), followed by mannosidosis (5 cases). No cases of fucosidosis were found. PMID- 2964539 TI - Atrial natriuretic peptide involvement in human platelet aggregability in vitro. AB - Atrial natriuretic peptide (ANP) counteracts the destabilization and aggregation of platelets which is induced by vasopressin, angiotensin, and adrenaline. PMID- 2964540 TI - [Results of PTCA following thrombolysis in acute myocardial infarct]. AB - The mortality in acute transmural myocardial infarction can be reduced by thrombolytic therapy administered within 6 hours after onset of symptoms. In patients with coronary angiography proven stenosis of less than 70% conservative therapy is recommended. In patients with one vessel disease PTCA in patients with suitable lesions should be used. Further studies have to elucidate, if the angioplasty has to be performed in the acute stage or on an elective basis. PTCA is recommended in patients with occluded coronary vessels and in patients with high grade stenosis and limited coronary blood flow. In patients with multi vessel disease aortocoronary bypass surgery is recommended to reduce high mortality. Long-term results demonstrate, that patients in whom PTCA or coronary bypass surgery was performed demonstrated the highest survival rate. PMID- 2964541 TI - [Drug therapy of coronary heart disease following thrombolysis and PTCA or bypass operation]. AB - Successful thrombolysis with reperfusion and salvage of jeopardized myocardium frequently necessitates invasive therapeutic strategies such as angioplasty or bypass surgery. Medical treatment in this situation has to consider the possibility of recurrence of infarction before invasive measures can be obtained. It must be specially directed towards preventing such an event, besides applying therapeutic measures in the acute and subacute phase after myocardial infarction. Several different therapeutic principles were considered: 1. In the days after thrombolysis with or without PTCA anticoagulants, mostly in the form of heparin, are used routinely in an attempt to prevent reocclusion and reduce thromboembolic complications. The latter indication seems to yield the most benefit of this type of drug. 2. Platelet-modifying drugs reduce the risk of reocclusion after thrombolysis and PTCA. The vein graft patency seems to be higher after aspirin treatment than after coumarine therapy. A reduction of coronary events was reported especially in patients with non Q-wave infarction, a situation particularly liable for reinfarction. 3. Calcium antagonists are used widely after PTCA to attenuate coronary spasm. No global beneficial effects have been observed after bypass surgery and in the therapy of acute myocardial infarction. Special indications e.g. for certain types for arrhythmias remain unchanged after thrombolysis, PTCA and bypass surgery. 4. Antianginal drugs with nitrates and betablockers are indicated to relieve symptoms in the acute phase. In chronic treatment, prevention of reinfarction and reduction of mortality was reported after the use of betablockers, the latter mainly by increasing the fibrillation threshold, thus reducing deaths due to arrhythmias.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964542 TI - Fibrinolytic agents and their effects on the haemostatic system. AB - Fibrinolytic agents used in intravenous thrombolytic therapy of myocardial infarction also exert, at effective dosages, significant side effects on the haemostatic system outside the direct resolution of the target thrombus. The side effects of the agents streptokinase and anisyolated plasminogen-streptokinase activator complex are larger than the side effects of the more fibrin-specific agents tissue-type plasminogen activator and pro-urokinase. An important difference between the two groups of compounds is that the major plasmin inhibitor alpha 2-antiplasmin is completely exhausted for the first two agents, but only in part of the cases for the fibrin-specific agents. The effects on various factors in the fibrinolytic cascade on the plasminogen activator, plasminogen-plasmin and fibrinogen-fibrin levels are reviewed concisely. PMID- 2964543 TI - GM2-gangliosidosis variant with altered substrate specificity: evidence for alpha locus genetic compound. PMID- 2964545 TI - Slavic ethnomedicine in the Soviet Far East. Part I: Herbal remedies among Russians/Ukrainians in the Sukhodol Valley, Primorye. AB - This paper is the first of a series of publications on Slavic ethnomedicine in the Soviet Far East. Field studies were carried out among Russian and Ukrainian residents of the Sukhodol Valley in Primorye, USSR. Primorye is the southernmost region of the Soviet Far East and the extreme southeastern area of the Soviet Union. The region is interesting because of its nature, climate and rich flora. Informants have shown broad knowledge of pharmacological properties of wild and cultivated plants of the area. The study has recorded 93 plants in 41 families. The use of six species had not been described before. PMID- 2964544 TI - C3 dependent, C5 independent immune complex glomerulopathy in the mouse. AB - This study examines the role of complement in a murine model of accelerated nonproliferative immune complex glomerulopathy. Two C5 deficient strains (DBA/2J and B10.D2oSnJ) as well as normocomplementemic mice consistently develop heavy proteinuria and glomeruli show loss of normal visceral epithelial cell architecture within 4 days of intravenous antigen administration. In contrast, animals depleted of C3 with cobra venom factor fail to develop proteinuria and retain discrete foot processes. Semiquantitative evaluation of antigen and antibody in glomeruli shows equivalent deposition in mice from all groups. The localization of these deposits, however, is different in C3-depleted mice. There is extensive accumulation of deposits along the subepithelial aspect of the glomerular basement membrane of normocomplementemic and C5 deficient mice while deposits in glomeruli of C3-depleted animals accumulate in the subendothelial region and do not cross the glomerular basement membrane. These data demonstrate that in this model, glomerular injury is dependent on complement components generated up thru C3 but not C5 or latter components. In addition, our data suggest that C3 is important in the movement of immune complexes across the glomerular basement membrane. Although the mechanism by which complement is mediating injury in this model is not known, it does not appear to involve an inflammatory cell infiltrate or the terminal complement components. PMID- 2964546 TI - Alzheimer's disease: an olfactory connection? AB - The density and distribution of senile plaques and neurofibrillary tangles were examined in the olfactory bulbs and tracts, amygdala and hippocampus of 28 patients with Alzheimer's disease, 13 with Down's syndrome and 60 non-demented patients of age range 6-84 years. In all three patient groups comparisons of incidence and severity over the three areas showed the amygdala to be the most commonly and most severely affected area by senile plaques, the hippocampus by neurofibrillary tangles, and the olfactory bulbs and tracts to be the least affected by both. These findings are discussed in relationship to the possibility that the olfactory tracts might provide a portal of entry to the brain for any putative pathogenic agent(s) that might be responsible for the induction of senile plaques and/or neurofibrillary tangles. PMID- 2964547 TI - T cell activation through different membrane structures (T3/Ti, T11, T44) and frequency analysis of proliferating and interleukin-2 producer T lymphocyte precursors in aged individuals. AB - It is well known that the proliferative responsiveness of T cells of aged subjects is depressed in both autologous mixed lymphocyte reaction (AMLR) and in PHA-induced cultures. In the present study we analyzed T cell activation through different stimulatory pathways (such as T3/Ti antigen receptor, T11 complex and T44 molecule). Moreover, we studied Interleukin-2 (IL-2) release performing a limiting dilution analysis of the proliferative capability of peripheral blood T cells, employing a high efficiency cloning technique. Our results demonstrate normal proliferation of T3-induced T cells in aged subjects, whereas T11- and T44 induced T cell proliferations are depressed in aged subjects. In addition, studies at clonal level reveal a normal percentage of IL-2 producer T cell in aged individuals. In conclusion, our data suggest that the T cell in aged subjects are normal in number, but they have a decreased capacity of lymphokine production. PMID- 2964548 TI - Effects of nortriptyline and its 10-hydroxy metabolite on plasma noradrenaline (NA) concentrations, heart rate and blood pressure during intravenous NA infusion. AB - Eight healthy subjects were randomly given placebo and equimolar doses of nortriptyline (NT) and E-10-hydroxy-NT (E-10-OH-NT). Two hours after oral intake of drug, noradrenaline (NA) was infused intravenously at three consecutive rates. Before infusion of NA, E-10-OH-NT significantly increased heart rate compared to NT (p less than 0.05) and placebo (p less than 0.01). During NA infusion, the active drugs caused non-significant tendencies to augmented increase of blood pressure and decrease of heart rate. Plasma NA concentrations increased significantly due to the infused NA but were not influenced by NT or E-10-OH-NT. This absence of drug effect may have been due to several simultaneously operating factors affecting plasma NA concentrations, e.g., modification of the rates of NA release and clearance by exogenous NA or drugs and competing elimination pathways for infused NA. After stopping the NA infusion, a non-significant tendency to a slower elimination of NA was found after both active drugs. PMID- 2964549 TI - Plasma levels of alpha-atrial natriuretic peptide in normal subjects under various postural conditions. AB - The present paper describes a radioimmunoassay, in conjunction with an extraction procedure on octadecasilyl silica cartridges, of alpha-atrial natriuretic peptide in human plasma and its application in normal subjects under various postural conditions. The sensitivity of this assay is 1.96 pg/ml. The plasma concentrations of alpha-atrial natriuretic peptide in 18 normal male subjects ranged from 10 to 55 pg/ml. This assay was shown to be reproducible as indicated by the low within- and between-assay coefficients of variation. The plasma alpha ANP concentration decreased during prolonged storage of the plasma at -20 degrees C or -80 degrees C and its level is also lower in hemolyzed plasma samples. The plasma alpha-ANP concentration is similar in standing and supine position but rose significantly when the subjects were tilted 20 degrees head-down. PMID- 2964550 TI - A guide to the Workers' Compensation Act, 1987. Workcover. PMID- 2964551 TI - Pharmacokinetics of buspirone as determined by ex vivo (3H)-DPAT binding. AB - Ex vivo (3H)-8-hydroxy-2-(di-n-propylamino)-tetraline ((3H)-DPAT) binding to the hippocampus has been utilized to determine the pharmacokinetic parameters of buspirone after i.v. (30 mumol/kg) and oral (100 mumol/kg) administration of this drug to rats. Intravenous buspirone rapidly penetrated the brain as demonstrated by a maximum inhibition of (3H)-DPAT binding at 1 min. Elimination of drug from the brain was biphasic, with a first component half-life of 24.8 min and a second component half-life of 96 min. Oral buspirone at 3 times the i.v. dose produced less than one-third the maximum inhibition of (3H)-DPAT binding compared to that observed with i.v. buspirone. The pharmacokinetic parameters of buspirone observed in the present study are in agreement with those reported previously. Thus, the ex vivo binding assay could be utilized to determine the bioavailability of the drug to the brain, and its duration of action. PMID- 2964553 TI - Effect of atrial natriuretic peptide on acetylcholine-induced release of corticotropin-releasing factor from rat hypothalamus in vitro. AB - Effect of rat atrial natriuretic peptide (rANP) on acetylcholine-induced release of corticotropin-releasing factor (CRF) from the rat hypothalamus was studied in vitro using perifusion method. Perifused acetylcholine at 100 and 1000 ng/ml evoked significant CRF release, whereas norepinephrine at 10, 100 and 1000 ng/ml did not show a definite effect on CRF release. Continuous administration of alpha rANP(1-28) (20ng/ml) inhibited the acetylcholine (100ng/ml)-induced CRF release. It is likely that ANP is involved in the regulation of CRF release. PMID- 2964552 TI - Effect of sodium ion on atrial natriuretic factor release from rat hypothalamic fragments. AB - The effects of Na ion and choline chloride on the release of atrial natriuretic factor (ANF) and growth hormone-releasing factor (GHRF) from rat hypothalamic fragments including the organum vasculosum of the lamina terminalis (OVLT) were examined in vitro. Although the release of ANF was stimulated by Na ion, choline chloride, and glucose in concentration-dependent manners, the release was more sensitive to a change in concentration of Na ion than to those of choline chloride and glucose. On the other hand, the change in Na ion concentration did not affect the release of GHRF. It can be therefore proposed that Na ion is the first candidate controlling ANF release from the brain tissue and that ANF in the hypothalamus and/or OVLT may play some role in the regulation of the Na ion and water balance in the central nervous system. PMID- 2964555 TI - [Work experience of the wall newspaper in the medical school]. PMID- 2964554 TI - [Functional obligations of the nurse in the pediatric pulmonology sanatorium]. PMID- 2964556 TI - [Immunological changes in patients with chronic idiopathic thrombocytopenic purpura]. PMID- 2964557 TI - Calcium transport mechanisms in epithelial cell membranes. AB - The data which we have reviewed suggest that, at steady state, the cytosolic [Ca2+] is determined by the balance between Ca2+ influx across both the brush border and the basolateral membrane, and Ca2+ extrusion through the basolateral membrane catalyzed predominantly by the calmodulin-activated Ca2+ ATPase. Na+ Ca2+ exchange does not appear to play an important quantitative role in calcium extrusion but may participate in the regulation of cytosolic calcium as a function of luminal sodium entry. Ca2+ transport systems in endomembranes are probably activated only during Ca2+ transients in the cell. Thus, the reticular high-affinity Ca2+ ATPase allows for accumulation of limited amounts of Ca2+ into the reticulum at normal cytosolic Ca2+ concentrations of 0.1-0.2 M; Ca2+ can be rapidly released from this pool by IP3 produced in response to hormonal stimulation, which results in a transient increase in cytosolic Ca2+ and stimulation of calmodulin-dependent enzymes. When Ca2+ entry is stimulated to levels which exceed the capacity of Ca2+ ATPase in the basolateral membrane, cytosolic [Ca2+] may rise above the mitochondrial set point and mitochondrial Ca2+ accumulation may occur. It remains to be investigated if the latter reaction occurs under physiological or only under pathological conditions. PMID- 2964558 TI - Differential effects of carbohydrate intake on cardiac myosin isoform expression in normal weanling and adult rats. AB - Dietary manipulations involving high carbohydrate feeding increase V1 cardiac myosin isoform expression in hormonally deficient rats. The purpose of this study was to determine if extremes in dietary carbohydrate availability could alter cardiac myosin isoform patterns in normal weanling and adult rats. Three and six weeks of dietary manipulations (either high or low carbohydrate diets) failed to change calcium-activated myofibril ATPase activity, calcium regulated myofibril ATPase activity, or the myosin isoform distribution in the adult. In contrast, a four week, high carbohydrate diet reduced calcium activated myosin ATPase activity by 33%, calcium regulated myofibril ATPase activity by 10%, and V1 isoform expression by 66% in weanling rats. Although the low carbohydrate diet caused no change in the myosin ATPase properties, it decreased V1 isoform expression by 17%. These results show that carbohydrate availability can alter cardiac myosin isoform expression in normal rats, but only at weanling age. The reason for this age-related contrast in response to dietary manipulations is unknown at this stage. The dietary manipulations may have acted directly on the heart by creating a state of malnutrition, or indirectly, by altering some developmental process which links maturation of the sympathetic nervous system with myosin isoform expression. PMID- 2964560 TI - The site of the earliest lesions of Alzheimer's disease. PMID- 2964559 TI - The effects of felodipine and bepridil on calcium-stimulated calmodulin binding and calcium pumping ATPase of cardiac sarcolemma before and after removal of endogenous calmodulin. AB - The Ca2+ channel blockers felodipine and bepridil are known to affect selectively functions of calmodulin. We studied their effects on calmodulin binding and ATPase activities of calmodulin-containing and calmodulin-depleted rabbit heart sarcolemma. Both drugs as well as the specific anti-calmodulin drug calmidazolium at a concentration of 50 microM, inhibited the Ca2+-stimulated calmodulin binding to calmodulin-depleted sarcolemma. Within the concentration range of 3 to 100 microM all three drugs also progressively inhibited Ca2+ pumping ATPase in calmodulin containing sarcolemma, although the enzyme was assayed at saturating Ca2+ (100 microM). The inhibitory potency of calmidazolium and bepridil, but not that of felodipine, increased when the membrane protein concentration in the ATPase assay was lowered. At low membrane protein concentration 30 microM calmidazolium completely blocked calmodulin-dependent Ca2+ pumping ATPase, whereas the inhibition caused by 30 microM felodipine or bepridil remained partially. A similar inhibition pattern of the drugs was found in the calmodulin binding experiments. Within a concentration range of 3 to 30 microM, all three drugs had negligible effects on the basal Ca2+ pumping ATPase which was measured in calmodulin-depleted sarcolemma. In conclusion, the characteristics of the anti calmodulin action of felodipine on the rabbit heart sarcolemmal Ca2+ pumping ATPase are not different from those of bepridil. Both drugs may inhibit the enzyme by interference with the Ca2+-stimulated binding of calmodulin. PMID- 2964561 TI - Compensation for disease--non-mining industry. PMID- 2964562 TI - A new natriuretic peptide in porcine brain. AB - Atrial natriuretic peptide (ANP), a hormone secreted from mammalian atria, regulates the homoeostatic balance of body fluid and blood pressure. ANP-like immunoreactivity is also present in the brain, suggesting that the peptide functions as a neuropeptide. We report here identification in porcine brain of a novel peptide of 26 amino-acid residues, eliciting a pharmacological spectrum very similar to that of ANP, such as natriuretic-diuretic, hypotensive and chick rectum relaxant activities. The complete amino-acid sequence determined for the peptide is remarkably similar to but definitely distinct from the known sequence of ANP, indicating that the genes for the two are distinct. Thus, we have designated the peptide 'brain natriuretic peptide' (BNP). The occurrence of BNP with ANP in mammalian brain suggests the possibility that the physiological functions so far thought to be mediated by ANP may be regulated through a dual mechanism involving both ANP and BNP. PMID- 2964563 TI - An ATPase with properties expected for the organelle motor of the giant amoeba, Reticulomyxa. AB - The rapid, vectorial, microtubule-associated transport of organelles is believed to be mediated by specific mechanochemical transducers. Recent studies of various metazoan cells have allowed the identification of novel microtubule-dependent translocator molecules capable of promoting microtubule gliding across glass surfaces and translocation of inert beads along microtubules. These translocators could be involved in force generation for directional organelle movements in vivo. Here we report the identification of a microtubule-binding protein with characteristics expected for an organelle translocator in the giant freshwater amoeba Reticulomyxa. This factor has an apparent relative molecular mass (Mr) of 440,000 (440K) and sediments at 20-22S in sucrose-density gradients. It binds to microtubules under conditions of ATP depletion, possesses an ATPase activity and is sensitive to ultraviolet-induced, vanadate-dependent cleavage. Although its pharmacological properties differ from those of axonemal dynein, it can be considered to be a variant of cytoplasmic dynein. The Reticulomyxa high-molecular weight protein (HMWP) promotes rapid, bidirectional movement of latex beads along Reticulomyxa microtubules in vitro at an average speed of 3.6 micron s-1. This protein, therefore, is a likely candidate for a microtubule-dependent motor. PMID- 2964564 TI - Effects of the selective 5-hydroxytryptamine uptake inhibitors paroxetine and zimeldine on EEG sleep and waking stages in the rat. AB - The effects of oral paroxetine and zimeldine on EEG sleep-waking phases in the rat were investigated over a wide dose range. To ascertain that at the doses used for the EEG studies paroxetine and zimeldine selectively affect the serotoninergic system, their effects on brain 5-hydroxytryptamine (5-HT), dopamine and noradrenaline were determined. It was found that paroxetine and zimeldine at doses of 1-18 mg/kg dose-dependently prolonged waking and shortened slow-wave sleep and paradoxical sleep. In the same dose range cortical 5-HT turnover was significantly reduced, whereas the other aminergic systems were not influenced. These results suggest that 5-HT uptake inhibitors increase vigilance in rats at oral doses which selectively stimulate the serotoninergic system. PMID- 2964565 TI - Clinical and neuropathologic assessment of severity in Huntington's disease. AB - Clinical records were evaluated for 163 Huntington's disease patients in whom postmortem brain specimens had been graded for degree of neuropathologic involvement in the striatum. Juvenile/adolescent onset (4 to 19 years of age) was associated with very severe neuropathologic involvement produced by an apparent rapid degenerative process. Cases of early (20 to 34 years) and midlife (35 to 49 years) onset had respectively less severe striatal involvement, suggesting a slower degenerative progression. High correlations among the grade of neuropathologic involvement, cell counts of neurons, and a rating of physical disability suggest that each represents a common underlying degenerative process of the disease. The relationship between the age at onset and the extent of neuropathologic involvement suggests that a single mechanism may determine both onset and rate of degenerative disease progression. PMID- 2964566 TI - Saccades in Huntington's disease: slowing and dysmetria. AB - Eye movements were recorded from 20 mildly affected patients with Huntington's disease (HD) who were divided into two groups, 10 patients with onset of symptoms before age 30 and 10 with onset of symptoms after age 30. In the younger onset group (HD less than 30), peak saccade velocities were low (less than 255 deg/sec for 20-deg saccades) in six of the 10 patients, whereas none of the 10 patients in the older onset group (HD greater than 30) had peak saccade velocities lower than 300 deg/sec. Latencies for volitional saccades were greater than normal in the HD greater than 30 group, but were normal for the HD less than 30 group. The ability to maintain steady fixation in the face of a distracting visual stimulus was decreased, to the same degree, in both groups of HD patients. In addition, 70% of the HD less than 30 group had an affected father, while 70% of the HD greater than 30 group had an affected mother. These findings suggest that the pathophysiology of the slow saccades, initiation deficit, and excessive distractibility in HD are different. PMID- 2964567 TI - Renal response to atrial natriuretic peptide in nephrotic syndrome. AB - In six patients with nephrotic syndrome of various aetiology, the increase in absolute and fractional sodium excretion (FENa) after a bolus injection of 100 micrograms alpha-human atrial natriuretic peptide (ANP) was not different from the effect in normal healthy subjects at comparable sodium levels. Glomerular filtration rate rose in normals as well as in patients. In two patients, however, baseline sodium excretion was very low and the natriuretic response to ANP was proportionally blunted. The high baseline sodium reabsorption and blunted response to ANP may be explained as due either to an intrinsic renal defect or to a circulatory hypovolaemia. The finding of a low plasma ANP in these two patients, however, suggests involvement of a hypovolaemic component. PMID- 2964568 TI - Irradiation inactivation studies of the dopamine D1 receptor and dopamine stimulated adenylate cyclase in rat striatum. AB - In frozen rat striatal tissue, exposed to 10 MeV electrons from a linear accelerator, the sizes of the dopamine (DA) D1 receptor and the DA-sensitive adenylate cyclase complex were determined using target size analysis. The number of D1 receptors (labelled by [3H]SCH 23390) declined monoexponentially with increasing radiation intensity, yielding a molecular weight (mol.wt.) of 80 kDa. Also the activity of the catalytic unit (C) of the adenylate cyclase (as measured by forskolin stimulation), decreased monoexponentially, however with a mol.wt. of 145 kDa. Both basal, DA- and fluoride (F-)-stimulated activity declined in a concave-downward fashion with a limiting mol.wt. of 134, 138 and 228 kDa, respectively. It was estimated that the basal and DA-stimulated activity originated from an enzyme complex with a mol.wt. of 325 kDa, a value close to the combined size of RGs and C. These data suggest that F- stimulation of the adenylate cyclase, which occurs by a Gs activation, does not cause dissociation of Gs into the alpha s and beta gamma subunits. Further, the DA-regulated adenylate cyclase apparently exists as a complex consisting of RGs and C; the mechanism of hormonal activation is a dissociation of C from this complex. PMID- 2964569 TI - Autoradiographic localization of dopamine D2 receptors in the rat brain using the new agonist [3H]N-0437. AB - The selective dopamine D2 agonist [3H]N-0437 was used to label dopamine receptors in vitro in slide-mounted rat brain microtome sections. The characteristics of the binding of [3H]N-0437 to tissue section were similar to those observed previously in membrane preparations and indicated that this ligand labels sites with the properties of a dopamine D2 receptor. The regional distribution of these receptors was examined by autoradiography and quantified by computer-assisted microdensitometry. The highest densities of [3H]N-0437 sites were observed in the nucleus caudate-putamen, accumbens, olfactory tubercle, island of Calleja and the glomerular layer of the olfactory bulb. Lower densities of binding sites were seen in stratum griseum superficialis of the superior colliculus, substantia nigra pars compacta and area ventral tegmental, entorhinal cortex and in the molecular layer of the 9th and 10th lobules of the cerebellum. Very low densities were seen in the neocortex and hippocampal formation. The density of [3H]N-0437 binding sites in the rat striatum are higher than those observed with other dopamine D2 [3H]agonists and comparable to those seen with [3H]antagonists. [3H]N 0437 is a new useful tool for the anatomical localization of dopamine D2 receptors in brain. PMID- 2964570 TI - Ureteral injury after laparoscopic tubal sterilization by bipolar electrocoagulation. AB - In bipolar electrocoagulation of the fallopian tubes, the electric current is between the tips of the forceps, which are visualized. Therefore, unnoticed burn injuries are uncommon. We describe a case of ureteral stenosis due to inadvertent burn injury, which was successfully managed conservatively. PMID- 2964571 TI - Medical detective must develop uniform workers' comp approach. PMID- 2964572 TI - [Malignant neoplasms with secondary, unspecified or unknown site--significance for cause of death statistics]. PMID- 2964573 TI - [Analysis of the classification of work disability diagnoses with ICD no. 714 based on rheumatologic follow-up]. PMID- 2964574 TI - [AIDS in sailors]. PMID- 2964575 TI - [Assessment of the vaccination status of students beginning school and of grade 4 students in the Rastatt, Baden-Baden and Buhl area]. PMID- 2964576 TI - [Are involuntary commitments decreasing due to impediments in the procedure?]. PMID- 2964577 TI - [Effect of social environment on diagnosis and therapy in surgery]. PMID- 2964578 TI - [Importation of infectious diseases by refugees and its significance for the diagnostic program of the public health office]. PMID- 2964579 TI - [Cause of death of young humans in a county district]. PMID- 2964580 TI - [Social workers in the changing times]. PMID- 2964581 TI - [Problems of readiness to help and competence of lay persons in emergencies]. PMID- 2964582 TI - [Parental guidance--status of health education and guidance of parents with infants and development possibilities from the viewpoint of the public health office]. PMID- 2964583 TI - [Degree of familiarity, frequency of use and success of alternative healing methods in Austria--results of a population survey]. PMID- 2964584 TI - [Visual acuity and refraction in medical students]. PMID- 2964585 TI - Corrective cosmetics adjunctive to the fields of ophthalmology and plastic surgery. AB - An effective, stable cosmetic cover creme useful for ophthalmologists is presented. In a series of 52 patients, there was a high degree of patient acceptance. If properly used, the creme is effective and involves minimal risk because it has been subjected to sensitivity, photosensitivity, comedogenic, and sun blocking tests. The author recommends this camouflage creme for adjunctive use. PMID- 2964586 TI - The intraocular penetration and retinal toxicity of teicoplanin. AB - We investigated the intraocular penetration and retinal toxicity of teicoplanin, a relatively new glycopeptide antibiotic with activity similar to vancomycin when used to inhibit staphylococci and other gram positive organisms, particularly Streptococcus faecalia. Topically administered teicoplanin penetrated poorly into the aqueous and vitreous in rabbit eyes. Subconjunctival injection of the drug yielded aqueous levels above the minimum inhibiting concentration (3.1 micrograms/ml) only at one hour after injection. In the vitreous, drug levels were above the mean inhibitory concentration at 30 minutes after the subconjunctival injection, but rapidly declined thereafter. The maximum nontoxic, single-dose, intravitreal injection was 750 micrograms/0.1 ml. Rabbits received 8 micrograms/ml of teicoplanin in an intravitreal infusion solution without demonstrable retinal toxicity. PMID- 2964587 TI - [United interdisciplinary classification of cranio-cerebral trauma]. PMID- 2964588 TI - Standing on their own two feet. PMID- 2964589 TI - Left-ventricular cavity dimensions in children with normal and dilated hearts. AB - Studies were carried out to find how left-ventricular length and length/diameter ratio relate to body size and degree of dilation. By use of M-mode and two dimensional echocardiography, diastolic cavity long axis (Led), diastolic cavity diameter (Ded), systolic cavity long axis (Les), systolic cavity diameter (Des), fractional L shortening (SFL), and fractional D shortening (SFD) were measured in children, adolescents, and young adults between two and 23 years of age, with body-surface area (BSA) between 0.5 and 2.1 m2 and with a variety of volume loads and SFD values. In normal subjects, Led/Ded was about 1.9. Regardless of age and pathology (in this age range), Led correlated consistently with BSA (Led = 3.9 + 3.2 BSA), indicating that the long axis changes rather little with pathological dilation. A plot of Led/Ded vs BSA/D2ed (in m2/cm2) formed a straight-line relation: Led/Ded = 0.77 + 16.4 BSA/D2ed. Similar relations were found for end systolic dimensions. End-systolic L/D ratio exceeded end-diastolic L/D ratio to a degree that depended on both end-diastolic L/D ratio and SFD:Les/Des = Led/Ded + (0.22 + 2.67 Led/Ded)(SFD)2. Relations like these may be useful in the interpretation of echocardiographic images. The results suggest that left ventricular L/D ratio may be influenced by myocardial anisotropy (dominance of hoop over meridional fiber orientation tending to promote prolate shape especially during systole) and external factors that antagonize extension of the long axis. PMID- 2964590 TI - Thirst impairment elicited by intraventricular administration of vasopressin antagonists. AB - To determine whether centrally released vasopressin influences thirst, observations of osmotic thirst threshold, osmotic load excretion and postloading restitution of plasma osmolality were made in dogs in control experiments and during infusion of AVP antagonists into the third ventricle. Significant elevation of osmotic thirst threshold was elicited by infusion of d(CH2)5AVP at a rate of 0.2-2.0 micrograms.min-1 and of d(Et2)AVP at a rate of 0.3 micrograms.min 1 (V1 antagonists, weak V2 agonists) as well as by administration of d(CH2)5[D Ile2,Abu4]AVP at a rate of 0.4 micrograms.min-1 (potent V2 antagonist, weak V1 antagonist). Administration of d(CH2)5AVP at a rate of 2.0 micrograms.min-1 was associated with a significant suppression of the postloading water intake and osmotic load excretion and with a delay in restitution of plasma osmolality. These findings indicate that centrally released vasopressin may participate in the control of thirst. PMID- 2964591 TI - Isolation of immunoreactive beta-endorphin-related and Met-enkephalin-related peptides from the posterior pituitary of the amphibian, Xenopus laevis. AB - Acid extracts of the posterior pituitary of the amphibian, Xenopus laevis, were analyzed with two heterologous region specific beta-endorphin RIAs. Following gel filtration chromatography and cation exchange chromatography four peaks of immunoreactivity were detected. All four peaks were detected with a N-acetyl specific beta-endorphin RIA. Peak I represented 92% of the total immunoreactivity isolated following cation exchange chromatography. This peak had a net positive charge at pH 2.5 of +1 and an apparent molecular weight of 1.4 Kd. Following reverse phase HPLC, Peak I fractionated into two peaks: Peak Ia and Peak Ib. Both peaks were detected with the N-acetyl specific beta-endorphin RIA and a Met enkephalin RIA, however, neither peak co-migrated with either Met-enkephalin or N acetyl-beta-endorphin(1-16). At present it is not clear whether Peak I is derived from pro-opiomelanocortin or one of the other opioid polyproteins. Peaks II, III, and IV represented 8% of the total immunoreactivity recovered following cation exchange chromatography. These peaks had net positive charges of +3, +4, and +5, respectively, and apparent molecular weights of 2.8, 3.2, and 3.5 Kd, respectively. These apparently N-acetylated beta-endorphin-sized forms are minor end products of the pro-opiomelanocortin biosynthetic pathway. PMID- 2964592 TI - [A case of extensive defect of the abdominal wall treated surgically by implanting the Dallop fascial prosthesis]. AB - A patient is described with an advanced tumor of the abdominal wall 15 x 10.5 x 10 cm in size, which grew into the left liver lobe and the transverse colon. The altered tissues were resected along with a wide margin of surrounding tissues. The defect in the abdominal wall was covered by the Dallop fascia prosthesis. There were reported no complications in the postoperative period. PMID- 2964593 TI - [Symptomatic and prognostic differences according to ethnic group in systemic lupus erythematosus. A controlled study of 3 populations]. AB - Systemic lupus erythematosus (SLE) has been found in all ethnic groups, but some of these groups--notably the black populations of the United States--seem to develop severe forms of the disease. We compared the signs and course of SLE in 20 black patients from the French West Indies, 20 patients of North African origin and 40 European Caucasians. At the onset of the disease, most of the West Indian and North African patients were living in France, and their social level was similar to that of the European patients. On the whole, our study confirmed that SLE is particularly severe in black populations. This severity is primarily due to renal involvement: 7 of the 13 renal biopsies we performed showed diffuse proliferative glomerulonephritis. In North African patients the severity of SLE was intermediate between that observed in West Indians and in European Caucasians. Five out of our 40 West Indian and North African patients died, as against only one female patient among the 40 European Caucasians. These differences seem to be ascribable to genetic factors rather than to environmental factors. PMID- 2964594 TI - [Intracorporeal concealment of narcotics. Experience of medicolegal emergencies at the Hotel-Dieu Hospital in Paris: 100 cases]. AB - One hundred cases of intracorporeal concealment of narcotic drugs were observed over a 6-month period at the medico-judicial emergency centre of the Hotel-Dieu hospital, Paris. Twelve cases concerned dealers who had ingested packs at the time they were arrested. The other subjects had been carrying the drug in their digestive tract, rectum or vagina for transportation. The illicit drugs thus transported were cocaine in 20 cases (ingestion), heroin in 65 cases (ingestion 7, rectal or vaginal cavity 58) and cannabis in 3 cases. Packs for sale being fragile, the dealers were particularly exposed to overdosage. Their management prior to intoxication was the same as that of acute ordinary drug poisoning, i.e. washing out of the stomach. In carriers, the risk of severe poisoning was associated with ingestion. The lack of antidote makes cocaine poisoning particularly dangerous. Surgical extraction is now reserved to cases with intestinal mechanical complications and to those increasingly rarer cases of fragile transportation packs. PMID- 2964595 TI - [Pharmacokinetics of netilmicin in cirrhotic patients with or without ascites]. AB - The pharmacokinetics of netilmicin after intramuscular injection (2 mg/kg) was investigated comparatively in cirrhotic patients with or without ascites, and in healthy subjects. In patients with ascites, the same pharmacokinetic parameters were measured after the ascites had been cured. Twenty-four hours after intramuscular injection, the residual levels in cirrhotic patients were moderately higher than in controls, showing that liver failure or ascites did not significantly modify the pharmacokinetics of netilmicin. Serum concentrations were bactericidal. The ascitic fluid level was lower than the therapeutic range, but it was sustained for nearly 24 hours after intraperitoneal injection (2 mg/kg). These results indicate that netilmicin may be administered to cirrhotic patients without peritoneal infection using the same regimen as in healthy subjects. The peritoneal route may be preferable in case of peritoneal infection. PMID- 2964596 TI - [Current pathogenetic data on mesangial glomerulonephritis with immunoglobulin A deposits]. AB - The pathogenesis of mesangial immunoglobulin A nephropathies was elucidated through the analysis of their clinical features and investigation of their biological characteristics. An alteration of the mucosal immune system is currently considered chiefly responsible for these diseases. A dysregulation of other compartments of the immune system is likely to enhance this abnormality and/or contribute to the persistence of immune complexes. Little is known, however, about the etiology of these renal diseases, and their treatment remains palliative or preventive. PMID- 2964597 TI - [Glomerulonephritis with IgA deposits is one of the most frequent causes of terminal renal insufficiency. Epidemiologic inquiry]. PMID- 2964598 TI - [Contribution of thoracic x-ray computed tomography to the diagnosis of esophageal perforation. A case]. PMID- 2964599 TI - [Urinary assay of copper containing superoxide dismutase in chronic glomerulonephritis]. PMID- 2964600 TI - [Radioimmunological assay of antimicrosomal antibodies in thyroid diseases]. PMID- 2964601 TI - [Efficacy of smectite in gastroesophageal reflux in the newborn infant]. PMID- 2964602 TI - [Hemolytic syndrome caused by myxoma of the right atrium]. PMID- 2964603 TI - [Ultrasensitive determination of thyrotropin in Graves' disease in remission]. PMID- 2964604 TI - [N-acetylaspartic aciduria: a new form of demyelinating encephalopathy]. PMID- 2964605 TI - [Immunology of polydermatomyositis]. PMID- 2964607 TI - [The genetics of narcolepsy in 1987]. PMID- 2964606 TI - [Efficacy of halofantrine in Plasmodium falciparum or Plasmodium vivax malaria in a resistance area (French Guiana)]. AB - Halofantrine (WR 171.669) is a phenanthrene methanol derivative effective against the multidrug resistant strains of Plasmodium falciparum. One hundred and one patients, 48 men and 53 women, 53 adults and 48 children (less than or equal to 12 years old) aged from 1.5 to 57 years were treated. Fifty-one patients received a single 16 mg/kg dose and 50 patients received 24 mg/kg/day in 3 doses at 6-hour intervals. Parasite counts with examination of both thin and thick smears were performed twice daily for 5 to 6 days following treatment, or until smears were negative for parasites for 24 hours, and then weekly for 4 weeks. Thirteen patients reported clinical side effects. Six treated patients had no parasites. One patient had mixed parasitemia. Eighty three patients had P. falciparum malaria, with mean parasitemias between 26,850 +/- 36,679 and 35,412 +/- 50,527 per cubic millimeter. Halofantrine was very effective in the two doses tested from 87.5 to 100 p. 100. Eleven patients had in vivo resistant strains; ten in vitro tests were successful and nine were resistant to chloroquine. Thirteen patients with P. vivax and a mean parasitemia of 13,858 +/- 10,835 per cubic millimeter were cured but 3 had a relapse 3 to 4 weeks after treatment. At the 2 dosage levels tested halofantrine proved highly effective in the treatment of malaria caused by resistant and sensitive strains to P. falciparum. PMID- 2964608 TI - [Epidemiology of cerebrovascular accidents in the young subject]. PMID- 2964609 TI - [Aortic valve replacement in persons over 75. 128 operated patients]. AB - Aortic valve replacements are performed in ever older subjects. In the surgical cardiovascular clinic of Rennes, 128 patients aged 75 or over (up to 85 years) and presenting with solitary or predominant (126 cases) aortic stenosis underwent aortic valve replacement between 1976 and 1985 inclusive. Pre-operative evaluation consisted, on principle, of non-invasive explorations. Myocardial protection was ensured by hypothermic cardioplegia. We used a mechanical (Bjork Shiley) prosthesis in the 19 patients operated upon before 1980, and a bioprosthesis (Carpentier-Edwards porcine, supra-annular type since 1983), in the 109 patients operated upon since 1980. The operative mortality rate was 8.6% (11 deaths). The survival curve was similar to that of a control population of the same age-group (survivors 75% at 4 years, operative mortality included). The quality of survival was remarkable since 96% of survivors were in NYHA stages I or II, the poor results being due to early or late cerebral vascular accidents. Advances in surgery (notably myocardial protection), anaesthesia and intensive care make it possible reasonably to operate upon very old patients, provided they have remained in good general and cerebral vascular condition. Non-surgical alternatives, such as percutaneous valvuloplasty, now used in elderly patients can only be reliable if results of similar quality and durability can be expected from them. PMID- 2964610 TI - [Hepatic protein synthesis of the kallikrein-kininogen-kinin system]. AB - Plasma prekallikrein and kininogens were assayed by specific enzymatic and immunological methods in cirrhotic patients with terminal liver failure and during oestrogenic impregnation. In cirrhotic patients plasma levels of these substances were significantly lowered and they correlated negatively with necrosis enzymes. A highly significant positive correlation was found between coagulation factor values and the levels of prekallikrein and kininogens. During oestrogenic impregnation the levels of the constituents of the kallikrein-kinin system were significantly increased when compared with reference values. These findings indicate that plasma concentrations of prekallikrein and kininogens are dependent upon liver synthesis capacity. PMID- 2964611 TI - [Analysis of intrathecal immunity]. AB - The occurrence of an intrathecal immune reaction, specific to the central nervous system, may reflect either a contemporary systemic immune reaction or a local process directly related to the disease observed. Analysis of this reaction proceeds in 3 stages: the intrathecal production of immunoglobulins is assessed; the specific antibody activity of the intrathecal immunoglobulins is compared with that observed in serum; possible variations in components of complement are investigated. These different data are derived from the examination of serum and cerebrospinal fluid samples obtained simultaneously. The results of the analysis help in determining the cause of the disease, enable the severity and potential prognostic value of the immune reaction to be assessed and provide precise information on the influence of treatment on the course of the reaction. Several examples (syphilis, AIDS, measles panencephalitis, multiple sclerosis) illustrate these results. PMID- 2964612 TI - [Vesical replacement by ileo-caecal graft detubulized after total prostato cystectomy]. AB - Following total prostato-cystectomy, the nearest substitute to a physiological reservoir is a constructed ileo-caecal pouch. The non-mesenteric sides of the caecum and ileum are incised on a length of 15 cm for each apex of the caecum which is anastomosed with the urethra, respecting the striated sphincter. The ureters are implanted into the caecal portion of the pouch. Owing to the length of the ileo-caecal mesenterium, this technique can be used in every case of prostato-cystectomy. Twelve cases followed up for 6 to 18 months are reported. PMID- 2964613 TI - [Digestive hemorrhages of Turner's syndrome. A case]. PMID- 2964614 TI - [Acute heart insufficiency during a pernicious malaria attack]. PMID- 2964615 TI - [Is glucagon a hyperglycemic agent without risk?]. PMID- 2964616 TI - [Is gastroesophageal reflux responsible for the episodes of nocturnal desaturation in chronic bronchitis patients?]. PMID- 2964617 TI - [Prolonged remission of Burkitt's lymphoma stage IV secondary to Hodgkin's disease]. PMID- 2964618 TI - [Do intravenous immunoglobulins have a place in the treatment of bullous pemphigoid?]. PMID- 2964619 TI - [Hyperglycemia in the initial stage of a cerebrovascular accident has a bad prognosis]. PMID- 2964620 TI - [French, European and international congresses and medical and scientific meetings from 1988 to 1990 inclusively]. PMID- 2964621 TI - [Ketoconazole therapy of Cushing's syndrome caused by adrenal polyadenomatosis]. PMID- 2964622 TI - [Use of a Fogarty probe in vascular perforations during subclavian catheterization]. PMID- 2964623 TI - [Chronic effect of tobacco on the partial pressure of transcutaneous oxygen]. PMID- 2964624 TI - [Consequences of hypophosphoremia in a case of severe meningitis]. PMID- 2964625 TI - [Adrenaline in cardiac arrest: aren't classical doses out of date?]. PMID- 2964626 TI - [Theophylline poisoning caused by interaction with high-dose ranitidine]. PMID- 2964627 TI - [Lithium-converting enzyme inhibitors: a combination to avoid?]. PMID- 2964628 TI - [Cancer of the prostate treated with 1 mg per day of diethylstilbestrol. Value of the determination of salivary free testosterone]. PMID- 2964630 TI - [Arterial compliance or elasticity?]. PMID- 2964629 TI - [Prolonged mydriasis caused by Lamaline]. PMID- 2964631 TI - [Extension to western Africa of the drug resistance of Plasmodium Falciparum]. PMID- 2964632 TI - [Perispinal morphine injections in the treatment of thoraco-abdominal cancer pain]. PMID- 2964633 TI - [Use of molar lactate in intracavitary electrophysiological study of a ventricular tachycardia]. PMID- 2964634 TI - [Dermal exposure to ethylenediamine in the petrochemical industry]. AB - During an examination of 47 workers engaged in ethylenediamine production subjective complaints of the skin were registered in 39% of them. Professional contact allergic dermatitis (26%) prevail over the contact irritative (6.5%) and lead very often to a change of employment for the operators. The epicutanic test with ethylenediamine and diethylene-triamine gives respectively 32 and 20% positiveness with high relative part of latent allergy. At compensated pH of the skin the sample for alkalic neutralization is prolonged in 42.9% of the workers. The ethylenediamine dermal absorption at concentrations in the air of the working area about MAC, is shown with methemoglobinemia registered in 15 operators. The raised methemoglobin in blood could be used with success as indirect test for exposure in hygiene-toxicological studies of workers exposed to ethylenediamine and aliphatic amines. PMID- 2964635 TI - [Physical work capacity of Bulgarian workers in the Komi ASSR]. AB - In view of assessing the physical fitness of Bulgarian men going to work in lumber production in the Komi ASSR (180 men of different professions) the physical working capacity was measured by bicycle ergometer with submaximum loading (50, 100 and 150 W). The maximum aerobic capacity is extrapolated after the nomogram of Astrand and the individual regressive lines. The results point out the mean values of physical working capacity assessed after oxygen consumption and pulse. Frequency at submaximum loading (at 150 W average for the group they are respectively 2, 12 dm3/min and 166 beating/min). The assessment of the physical working capacity after the maximum oxygen consumption is also about the mean (2.7 dm3/min). The representatives of the so called sitting professions (employees, drivers) have lower aerobic capacity then those engaged mainly in physical work (machine building, construction workers). In comparison with literature data the examined persons have lower working capacity for the corresponding age. Juxtaposing the results of the mean data for physical working capacity with the high energy losses in the lumber industry imposes measures for raising the physical working capacity of the Bulgarian workers departing for the Komi ASSr, by effective training system, improving the work and rest regimen, professional selection by examining the physical working capacity etc. PMID- 2964636 TI - 5-Fluorodeoxyuridine as an alternative to the synthesis of mixed hybridization probes for the detection of specific gene sequences. AB - Synthetic complementary oligonucleotides are useful hybridization probes for the detection of mRNAs and genes encoding proteins for which only a partial amino acid sequence is known. Usually this involves the synthesis of mixtures of oligonucleotides complementary to all possible bases in degenerate positions of codons. As an alternative we have prepared and characterized a series of unique oligonucleotides containing a pyrimidine analog, 5-fluorodeoxyuridine (F). Thermodynamic parameters and the melting temperatures of hybrid duplexes containing A.F and G.F base pairs showed that they are considerably more stable than duplexes containing A.T and G.T base pairs. The stability of a duplex decreased linearly with the number of mismatches introduced at positions at least a codon apart. A 5-fluorodeoxyuridine-substituted oligonucleotide cDNA detects rat liver pyruvate carboxylase mRNA on a RNA gel blot with a dissociation temperature only 10 degrees C below the measured melting temperature in solution. We suggest that the complexity of oligonucleotide cDNAs used for screening gene libraries can be reduced by the design of single hybridization probes containing the substituted bases--5-fluorodeoxyuridine to pair with adenosine or guanosine, guanosine to pair with cytidine or thymidine, and deoxyinosine to pair with adenosine or cytidine at positions of codon degeneracy--and still retain near maximum stability of hybrid duplexes. PMID- 2964637 TI - Suppression of contractile force in muscle fibers by antibody to myosin subfragment 2. AB - Polyclonal antibody directed against the subfragment-2 region of myosin was purified by affinity chromatography. Skinned muscle fibers that had been preincubated with antibody were able to sustain only 7% of the active isometric force generated by control fibers. The effect of antibody on force production could not be accounted for by inhibition of ATP turnover. PMID- 2964638 TI - Gene encoding the human beta-hexosaminidase beta chain: extensive homology of intron placement in the alpha- and beta-chain genes. AB - Lysosomal beta-hexosaminidase (EC 3.2.1.52) is composed of two structurally similar chains, alpha and beta, that are the products of different genes. Mutations in either gene causing beta-hexosaminidase deficiency result in the lysosomal storage disease GM2-gangliosidosis. To enable the investigation of the molecular lesions in this disorder and to study the evolutionary relationship between the alpha and beta chains, the beta-chain gene was isolated, and its organization was characterized. The beta-chain coding region is divided into 14 exons distributed over approximately 40 kilobases of DNA. Comparison with the alpha-chain gene revealed that 12 of the 13 introns interrupt the coding regions at homologous positions. This extensive sharing of intron placement demonstrates that the alpha and beta chains evolved by way of the duplication of a common ancestor. PMID- 2964639 TI - arcA (dye), a global regulatory gene in Escherichia coli mediating repression of enzymes in aerobic pathways. AB - In Escherichia coli the levels of numerous enzymes associated with aerobic metabolism are decreased during anaerobic growth. In an arcA mutant the anaerobic levels of these enzymes are increased. The enzymes, which are encoded by different regulons, include members that belong to the tricarboxylic acid cycle, the glyoxylate shunt, the pathway for fatty acid degradation, several dehydrogenases of the flavoprotein class, and the cytochrome o oxidase complex. Transductional crosses placed the arcA gene near min O on the chromosomal map. Complementation tests showed that the arcA gene corresponded to the dye gene, which is also known as fexA, msp, seg, or sfrA because of various phenotypic properties [Bachmann, B. (1983) Microbiol. Rev. 47, 180-230]. A dye-deletion mutant was derepressed in the aerobic enzyme system. The term modulon is proposed to describe a set of regulons that are subject to a common transcriptional control. PMID- 2964640 TI - Human and rat mast cell high-affinity immunoglobulin E receptors: characterization of putative alpha-chain gene products. AB - We have cloned and determined the entire nucleotide sequence of cDNAs corresponding to the putative alpha subunits of the human and rat mast cell high affinity IgE receptors. Both human and rat cDNAs encode an NH2-terminal signal peptide, two immunoglobulin-like extracellular domains (encoded by discrete exons), a hydrophobic transmembrane region, and a positively charged cytoplasmic tail. The human and rat alpha subunits share an overall homology with one another and the immunoglobulin gene family, suggesting that they arose from a common ancestral gene and continue to share structural homology with their ligands. In addition, the rat gene is transcribed into at least three distinct forms, each of which yields a somewhat different coding sequence. PMID- 2964641 TI - Temporal relationships of the anti-inflammatory effect of etodolac in the adjuvant arthritic rat. AB - Using the curative model of adjuvant arthritis, adult male Sprague-Dawley rats were treated with vehicle or etodolac (1, 3, and 8 mg/kg/day, po) for 9 days. Rats were sacrificed after 1, 2, 4, or 9 daily doses, and paw volume, PGE2 concentrations, and N-acetyl-beta-D-glucosaminidase (NAG) activity were determined in the left adjuvant-injected hindpaws. All three doses of etodolac caused a significant decrease in PGE2 concentrations after the first dose, and the decreases persisted for 2, 4, and 9 days of treatment, respectively. In rats given four daily doses of 3 and 8 mg/kg/day of etodolac, the paw volume was significantly decreased by about 50%, compared with that of the arthritic controls. A significant decrease in NAG activity was observed only after nine daily doses of 8 mg/kg/day etodolac. The sequence of anti-inflammatory events manifested following etodolac treatment would appear to be an initial inhibition of PGE2 synthesis, followed by resorption of fluid, and then by a reduction in macrophage infiltration. PMID- 2964642 TI - Effect of hemorrhage on plasma atriopeptin levels in conscious dogs. AB - An increase in atrial pressure has been shown to cause an increase in the concentration of atrial peptides (atriopeptin) in plasma. We therefore hypothesized that a reduction in atrial pressure would decrease the concentration of atriopeptin in plasma. In formulating this hypothesis we assumed that changes in the concentration of other circulating hormones or changes in cardiac nerve activity during hemorrhage would not affect the secretion of atriopeptin. To test the hypothesis, we bled sham-operated conscious dogs at a rate of 0.8 ml.kg-1.min 1 to decrease right and left atrial pressures. Hemorrhage was continued until a total of 30 ml of blood per kilogram body weight had been removed. Identical experiments were performed on conscious cardiac-denervated dogs. The concentration of plasma atriopeptin was decreased in each group of dogs after 10 ml of blood per kilogram of body weight had been removed, but the decrease achieved statistical significance only in the cardiac-denervated dogs. Further hemorrhage, however, produced no further decreases in circulating atriopeptin in either group even though atrial pressures continued to decline as more blood was removed. A comparison of the atriopeptin response to hemorrhage revealed no significant difference between the sham-operated and cardiac-denervated dogs, thus providing no evidence for a specific effect of cardiac nerves on atriopeptin secretion during hemorrhage. Our results demonstrate that the relationship between atrial pressure and plasma atriopeptin that has been observed repeatedly during atrial stretch is not evident during relatively slow, prolonged hemorrhage. There is, however, a small decline in circulating atriopeptin during the initial stage of hemorrhage that could be of biological significance. PMID- 2964643 TI - The effect of angiotensin II and ADH on the secretion of atrial natriuretic factor. AB - Studies in intact animals have suggested that angiotensin II (AII) and antidiuretic hormone (ADH) increase the plasma concentration of atrial natriuretic factor (ANF). The purpose of these studies was to examine the effects of AII and ADH on ANF secretion in a rat heart-lung preparation under conditions where aortic pressure could be regulated and other indirect effects of these hormones eliminated. ANF secretion was estimated as the total amount of ANF present in a perfusion reservoir at the end of each 30-min period. A pump was used to deliver a fluorocarbon perfusate to the right atrium at rates of either 2 or 5 ml/min. In a time control series where venous return was maintained at 2 ml/min for three 30-min periods ANF secretion was 672 +/- 114, 794 +/- 91, and 793 +/- 125 pg/min (n = 6, P greater than 0.05). When venous return was increased from 2 to 5 ml/min ANF secretion increased from 669 +/- 81 to 1089 +/- 127 pg/min (P less than 0.01). The addition of AII to the perfusate in concentrations of 50, 100, or 200 pg/ml (n = 6 in each group) had no significant effect on basal ANF secretion or the ANF response to increasing venous return. Similarly, the addition of ADH to the perfusate in concentrations of 5, 25, or 100 pg/ml had no significant effect on ANF release from the heart. These results suggest that the ability of AII and ADH to increase plasma ANF concentration in vivo may be due to the effects of these hormones on right or left atrial pressure. PMID- 2964644 TI - Characterization of intracellular Ca2+-stores in rat duodenal epithelium. PMID- 2964645 TI - Calcium oxalate-loaded sarcoplasmic reticulum vesicles from developing skeletal muscle are highly differentiated. PMID- 2964646 TI - Molecular basis of calcium and phosphate extrusion in renal and intestinal epithelium. PMID- 2964647 TI - Abnormal regulation of calcium transport in renal basolateral plasma membranes of Milan hypertensive rats. PMID- 2964648 TI - Measurements on the activity of the plasma membrane Ca pump ATPase in human hypertension. PMID- 2964649 TI - Chemistry of hematoporphyrin-derived photosensitizers. PMID- 2964650 TI - Studies on the structure of porphyrins contained in Photofrin II. PMID- 2964651 TI - The structure of the active material in hematoporphyrin derivative. PMID- 2964652 TI - Pulsed radiation studies of photodynamic sensitisers: the nature of DHE. PMID- 2964653 TI - Transport of light in tissue in photodynamic therapy. PMID- 2964654 TI - Photosensitizing properties of bacteriochlorophyllin a and bacteriochlorin a, two derivatives of bacteriochlorophyll a. PMID- 2964655 TI - Mitochondrial photosensitization by Photofrin II. PMID- 2964656 TI - Quantitative analysis of intracellular behaviour of porphyrins. PMID- 2964657 TI - Dihematoporphyrin ether-phospholipid interactions. The roles of surface charge and lateral phase separations. PMID- 2964658 TI - Dark effects of hematoporphyrin derivative on lactate dehydrogenase activity and distribution in HeLa cells: cytochemical evaluation. PMID- 2964659 TI - Growth delay studies of the response of V-79 multicell spheroids exposed to DHE and red light. PMID- 2964660 TI - The effect of porphyrin photosensitizers on the toxicity of 1,3-bis(2 chloroethyl)-1-nitrosourea in C57BL/6J mice. PMID- 2964661 TI - Photodynamic therapy of human bladder cancer cells in vitro correlated with cellular fluorescence levels of Photofrin-II. PMID- 2964662 TI - A fluorescence imaging device for endoscopic detection of early stage cancer- instrumental and experimental studies. PMID- 2964663 TI - Quantitative in vivo measurement of the fluorescent components of Photofrin II. PMID- 2964664 TI - Hematoporphyrin derivative uptake by atheroma in atherosclerotic rabbits: the spectra of fluorescence from hematoporphyrin derivative demonstrated by an excimer dye laser. PMID- 2964665 TI - Drug and light dose dependence of photodynamic therapy: a study of tumor and normal tissue response. PMID- 2964666 TI - Photodynamic therapy in the treatment of pancreatic carcinoma: dihematoporphyrin ether uptake and photobleaching kinetics. PMID- 2964667 TI - A test of different photosensitizers for photodynamic treatment of cancer in a murine tumor model. PMID- 2964668 TI - Ratioing fluorometer probe for localizing carcinoma in situ in bronchi of the lung. PMID- 2964669 TI - Skin photosensitivity: duration and intensity following intravenous hematoporphyrin derivates, HpD and DHE. PMID- 2964670 TI - [Adaptive reactions of the heart in arterial hypertension]. PMID- 2964671 TI - Behavioural specificity of 8-OH-DPAT-induced feeding. AB - 8-Hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT) has been reported to increase food intake. This effect was confirmed in rats consuming plain or sweetened wet mash. However, the consumption of a 35% sucrose solution or of a range of other flavoured liquids was enhanced marginally if at all by a single low dose of 8-OH DPAT, and the drug actually decreased the consumption of a high protein liquid diet. 8-OH-DPAT did, however, increase gnawing on a wooden block. These results suggest that 8-OH-DPAT-induced "hyperphagia" may to some extent be secondary to the elicitation of chewing behaviour. 8-OH-DPAT also caused certain other behavioural changes, including a suppression of grooming. PMID- 2964672 TI - The 5-HT1A agonist 8-OH-DPAT increases consumption of palatable wet mash and liquid diets in the rat. AB - The 5-HT1A agonist 8-OH-DPAT, at a dose of 30 micrograms/kg, enhanced the consumption of sweetened wet mash and sweetened milk in non-deprived rats. In partially satiated rats, sensitivity to the hyperphagic effect of 8-OH-DPAT on wet mash intake was substantially increased, so that the minimally effective dose was reduced to 3 micrograms/kg. Similarly, 8-OH-DPAT was more efficacious in increasing milk intake in satiated rats. Thus, 30 and 40 micrograms/kg 8-OH-DPAT produced a 4-fold increase of milk intake in completely satiated rats compared to a 2-fold increase in partially satiated animals at a dose of 30 micrograms/kg. The increased intake of liquid and wet mash diets observed after treatment with low doses of 8-OH-DPAT argues against the involvement of non-specific gnawing in the increased consumption of solid food produced by the drug. Rather, the data suggest that 8-OH-DPAT may specifically stimulate appetite by counteracting a tonic serotonergic inhibition of feeding. The present experiments also showed that 8-OH-DPAT attenuates fenfluramine-induced anorexia which is thought to depend on increased serotonergic neurotransmission. PMID- 2964673 TI - The midaortic syndrome: diagnosis and treatment. AB - The midaortic syndrome is an unusual entity seen in children and adolescents. It is characterized by severe narrowing of the abdominal aorta with progressive involvement of the renal and visceral branches. Eleven patients (aged 5 months to 15 years) suspected of having midaortic syndrome were examined preoperatively and postoperatively. All patients had hypertension and were examined with midstream aortography. All aortograms showed a smooth, segmental stenosis of the abdominal aorta and severe bilateral proximal renal artery stenosis. In three patients, percutaneous transluminal angioplasty of the renal artery was attempted, two preoperatively and one for a postoperative stricture. None showed long-term success, presumably due to the progressive nature of the disease. Grafts were surgically placed in ten patients and produced successful results in nine. PMID- 2964674 TI - Prosthetic arterial graft occlusion: a complication of tourniquet use during arteriography. AB - Tourniquet-induced reactive hyperemia, generally considered a safe procedure for enhancing visualization of the arteries in the distal lower extremity, caused acute occlusion of a Dacron arterial graft in one patient. This rare complication can be avoided through use of an alternative such as a chemical vasodilator in patients with susceptible synthetic vascular grafts. PMID- 2964675 TI - Atrial septal defect in older adults: atypical radiographic appearances. AB - Atrial septal defect (ASD) is the most common congenital cardiac anomaly manifested in adulthood. Clinical and radiographic features are well defined in patients less than 30 years of age. In older patients, however, the clinical symptoms are often atypical, and the auscultatory findings may be misleading. The radiographic manifestations in older patients have not been well described. In the current study, of 70 patients over the age of 50 years with proved ASD, 21 (30%) had atypical radiographic features, including apparently normal vascularity, left atrial enlargement, pulmonary venous hypertension, and pulmonary edema. In a control group of 70 younger patients with ASD, only 5.7% had atypical findings. The development of pulmonary venous hypertension and pulmonary edema in older patients was associated with smaller defects and a higher prevalence of mitral valve disease, left ventricular dysfunction, and pulmonary arterial hypertension than seen in older patients with typical radiographic findings. PMID- 2964676 TI - Flow reversal in the normal carotid bifurcation: color Doppler flow imaging analysis. AB - One hundred carotid bifurcations in young asymptomatic volunteers were examined with color Doppler flow imaging in an attempt to determine the frequency, configuration, magnitude, and duration of flow reversal. Areas of flow reversal were identified in all but one bifurcation. Just proximal to the bifurcation, flow reversal was typically located peripherally, opposite the origin of the external carotid artery. Distal to the bifurcation, flow reversal in the internal carotid artery typically assumed a linear configuration in the center of the vessel, extending from the superficial to the deep wall. Variations of these patterns were common. On the average, flow reversal occupied 33% of the lumen of the carotid bulb and extended for a distance of 14 mm. The average duration of flow reversal was 22% of the total cardiac cycle, with reversal typically occurring at peak systole and being replaced by static blood during diastole. These findings indicate that flow reversal in the carotid bifurcation is a normal phenomenon with a variable configuration, magnitude, and duration. PMID- 2964677 TI - Intestinal angina: percutaneous transluminal angioplasty of the celiac and superior mesenteric arteries. AB - Percutaneous transluminal angioplasty (PTA) was performed in ten patients with chronic ischemia of the mesenteric arteries and stenoses of the celiac artery and/or the superior mesenteric artery (SMA). PTA was performed on 14 occasions, with attempted dilation of 19 arteries. PTA was technically unsuccessful in two patients. Seventeen of 19 arteries (90%) were successfully dilated in eight patients, resulting in relief of symptoms that lasted from 6 to 24 months. Symptoms recurred in five patients, and redilation in three of them relieved the symptoms. Three patients, one of them following redilation, remained asymptomatic 7-9 months following PTA. An asymptomatic intimal dissection of the SMA was the only complication attributable to the procedure. PTA of celiac and SMA stenoses is an alternative to surgical revascularization and can be repeated if symptoms recur. PMID- 2964678 TI - Gender differences in variables associated with psychosocial adjustment to a burn injury. AB - The purpose of this study was to determine whether variables associated with psychosocial adjustment to a burn injury vary by gender. Male and female burned subjects (N = 260) were compared on their functional disability, disfigurement, coping responses, social resources, and psychosocial adjustment to a burn injury. Both men and women had adjusted psychosocially to their burn injury. Less functional disability (r = .57, p less than .001) for men and greater problem solving (r = .57, p less than .001) for women were the most important variables in explaining psychosocial adjustment to a burn injury. In the future, researchers need to be cognizant of gender differences and consider men and women as separate populations. PMID- 2964679 TI - [Arrhythmogenic dysplasia of the right ventricle. Evaluation of 7 cases using computerized tomography]. AB - Right ventricular arrhythmogenic dysplasia is a rare cardiomyopathy which involves the right ventricle either totally or partially. Up to now diagnoses have been based on Ecg, hemodynamics, angiography and echocardiography. This paper deals with the first 7 patients examined also by means of Computed Tomography. The CT picture is well defined and rather accurate. The most important elements are: total (6/7) or partial (1/7) enlargement of the right ventricle; thinning of right ventricular myocardium (6/7); marked increase in subepicardial fat limited to the right ventricular wall (7/7); right ventricle hypokinesia (7/7). A good correlation exists between CT, echographic and traditional methodologies findings. Because of its densitometric evaluation, CT is much more precise in the demonstration of fatty degeneration. Moreover, it can be very useful in differentiating intracardiac thrombi from hypertrophic papillary muscles and trabeculae. PMID- 2964680 TI - [Behavior of the ABO(H) blood group antigens of the urothelial surface in bladder tumors]. PMID- 2964681 TI - [Therapeutic modes in obstructions due to colon cancer]. PMID- 2964682 TI - [Benign stomach tumors of mesenchymal origin]. PMID- 2964683 TI - [Relationship of staging and postoperative survival in colon cancer]. PMID- 2964684 TI - [Residual lithiasis and recurrent lithiasis of the common bile duct. Diagnosis and treatment]. PMID- 2964685 TI - [Importance of the morphofunctional characteristics of the cuneometatarsal articulation in establishing indications for the surgical technic in the complex foot. A computer analysis]. PMID- 2964686 TI - [Recurrent perforated peptic ulcer following esophagoplasty]. PMID- 2964687 TI - [Cervical esophageal diverticulum exiting into the mediastinum. Its resolution by a double thoracic-cervical approach]. PMID- 2964688 TI - [Emergency therapy in upper digestive hemorrhages in patients with liver cirrhosis]. PMID- 2964689 TI - [Early and late complications of extracted intraocular foreign bodies]. PMID- 2964690 TI - [Leukocyte adherence inhibition (LAI) test in the diagnosis and prognosis of malignant melanoma of the eye]. PMID- 2964691 TI - [Diabetic retinopathy in myopia]. PMID- 2964692 TI - [Ophthalmologic emergencies represented by phosphenes and muscae volitantes]. PMID- 2964693 TI - [Kinetic visual acuity as a function of age, fatigue, training and natural selection of female textile workers at various work stations]. PMID- 2964694 TI - [Adverse effects of drugs on the visual capacity while driving]. PMID- 2964695 TI - [Ten years' experience in the implantation of artificial lenses]. PMID- 2964696 TI - [Current aspects of closed-angle glaucoma]. PMID- 2964697 TI - [Approach to the Cluj-Napoca Ophthalmologic Clinic in the treatment of the surgical treatment of concomitant strabismus]. PMID- 2964698 TI - [Drugs and the eye. II: Ocular reactions to anti-inflammatory analgesics]. PMID- 2964699 TI - [Progressive hemiatrophy of the face--Parry-Romberg syndrome]. PMID- 2964700 TI - [Difficulties in the early diagnosis of craniopharyngioma]. PMID- 2964701 TI - [Methodological communication regarding current clinical and therapeutic orientations in ocular tuberculosis (II)]. PMID- 2964702 TI - [Laryngotracheobronchial changes following long-term oro- or naso-tracheal intubation. II. Anatomopathological aspects]. PMID- 2964703 TI - [Intra-labyrinthine footplate fragments. A complication in surgery for deafness]. PMID- 2964704 TI - [Latent maxillary sinusitis in children]. PMID- 2964705 TI - [Dyscratic hemorrhage at the level of the airways]. PMID- 2964706 TI - [Lessons learned from the care of patients with unusual intracavitary foreign bodies]. PMID- 2964707 TI - [Rhinoliths--calculi of the antrum]. PMID- 2964708 TI - [Late neurologic manifestations in malignant pathology of the nasal sinus and nasopharynx]. PMID- 2964709 TI - [Complicated latent otomastoiditis in 0-3 year-old children, on the basis of 12 cases hospitalized in 1978-1985]. PMID- 2964710 TI - [Observations on 2 cases of osteoma of the external auditory canal]. PMID- 2964711 TI - [Current problems of psychiatric institutions and the zonal therapeutic system]. PMID- 2964712 TI - [Subarachnoid hemorrhage (multidimensional study)]. PMID- 2964713 TI - [Neurosurgical operations performed with the CO2 laser]. PMID- 2964714 TI - [Mental disorders associated with hyperkinetic syndromes in children]. PMID- 2964715 TI - [The epidemiology of depression]. PMID- 2964716 TI - Dracontiasis in the Igwun River Basin, Nigeria: its distribution, epidemiology, and transmission dynamics. AB - In a cross-sectional dracontiasis survey of the Igwun Basin, Nigeria, 879 persons from seven communities were examined. Ninety-five (10.8%) had dracontiasis blisters and ulcers. Male prevalence of 14.9% was significantly higher (P less than 0.001) than the female prevalence of 6.9%. Infection increased with age, rising from 2% in 1-4 years old children to 15.8% in the 35-44 years old adults. Sixty-five percent of infected males had blisters, 34.4% had ulcers. Among females, 48.4% had blisters, 51.6% had ulcers. Among the males, blisters and ulcers showed seasonal variations, with 80% cases of blisters between November and April, and 20% between June and October. Seasonal morbidity pattern in females did not show any significant variation (P greater than 0.20). The density of cyclopoid copepods increased in the dry season in relation to decreasing rainfall. The highest cyclops count was 18 cyclops per litre of water recorded between January and February, while the lowest was zero recorded in July in some areas. The infection rates of cyclops rose from 10% in November to a peak value of 20% between January and February, with the lowest rate of 5% in July. Larval burden of cyclops also rose from 3L3's/cyclops in June, to 5L3's/cyclops in January. Dracontiasis appears to be a relatively new disease in the area, and its spread has been enhanced by the interaction of human behavioural, and parasite vector ecological factors which are seasonally modulated. PMID- 2964717 TI - Motor depression as a consequence of inescapable shocking and a reactivity to the electric shocks: the acute tranylcypromin treatment. PMID- 2964718 TI - Effect of cystamine and 5-methoxytryptamine on the course of experimental bacteriemia in irradiated mice. PMID- 2964719 TI - Life conditions of persons with and without low-back pain. AB - A comparison of life conditions (such as dwelling, health, leisure-time activities, education, occupation, and work characteristics) was made between age and sex-stratified groups of persons with and without low-back pain (BP). The primary data were collected from a random, geographically standardized 1:1,000 sample of the Swedish population including 2,872 citizens aged 30-59 years. There were no differences with respect to dwelling, height, weight, habits of physical exercise, other non-occupational activities, and frequency of unemployment. In the group of 50-59 year old persons, the males with BP smoked more than those without. In the same age-group of females, and in the younger male groups there were no differences in smoking habits. Physically heavy, monotonous, and repetitive work was more frequent among the subjects with BP. The relative number of highschool-educated was lower among males--in the youngest age group--with BP. The back-healthy persons evaluated their state of health as better and their need of medical care as less than the persons with BP. PMID- 2964720 TI - [Eosinophilic granulocytes in tracheobronchial secretions of horses: evidence of parasitic lung disease?]. PMID- 2964722 TI - [Needs of a rural population in relation to sanatoria and preventoria]. PMID- 2964721 TI - Physical disability and social liminality: a study in the rituals of adversity. AB - Sociological research on the disabled has for the past 25 years made extensive use of a social deviance model to characterize the status of the physically impaired. The present article, which is based on a three-year anthropological investigation of the social relations of paraplegics and quadriplegics in the New York metropolitan area, argues that there are shortcomings in the deviance model and offers, instead, a model taken from the anthropological study of ritual. The disabled are viewed as being in a 'liminal' state, as in the liminal phases of rites of passages. They are persons having an undefined status: they are neither ill nor well, neither socially alive and active nor socially expunged and removed. The status of the disabled in American society and the symbolism of disability in American culture are reexamined within this framework. This perspective is extended to other types of deep adversity, such as acute loss of income and status or catastrophic illness. PMID- 2964723 TI - [Organizational aspects of the prevention and treatment of cardiovascular diseases]. PMID- 2964724 TI - [Anti-alcohol education of population groups of various ages and professions]. PMID- 2964725 TI - Comparison of clinical and objective methods of assessing trunk muscle strength- an experimental approach. AB - An objective method was used to evaluate the validity of the sit-up, prone trunk extension, and double leg lowering clinical tests. Normal men and women and patients with a history of backache served as subjects. The vast majority of the subjects were able to perform the Grade 1 level, which was the highest resistance imposed by the sit-up and prone extension tests. The double leg lower was superior to the other two clinical tests but was able to identify only broad differences in strength capability. In the main, the subjective clinical tests were poor discriminators and seemed to lack the range of resistance necessary to be definitively useful as methods of assessing trunk muscle strength and may not provide appropriate resistance for improving trunk muscle strength in most patients. PMID- 2964726 TI - Analysis of strength tests and resistive exercises commonly used for low-back disorders. AB - Using a modeling approach the conventional sit-up, the double straight-leg lowering, and prone trunk extension, the authors have shown these to have limitations as tests of maximum trunk flexion and extension strength and as modes of resistive exercise. These maneuvers are poor discriminators of trunk muscle strength, and they lack the range of resistance necessary to cover the spectrum of human trunk muscle strength capability. Use of manually handled weights can improve the above clinical tests and resistive exercise methods. As a result of selective positioning of the weights, variable resistance above and below that provided by the body segments alone can be achieved. The partial sit-up may be appropriate only for patients with significant trunk flexion weakness, and the straight head-trunk sit-up may be useful as a more strenuous test and resistive exercise. Rationale and derivations of the rigid-body equilibrium equations, as well as some of the limitations and assumptions associated with the modeling approach, are presented. PMID- 2964727 TI - The activity of individual trunk muscles during heavy physical loading. AB - The myoelectric activity of ten trunk muscles were recorded, using intramuscular electrodes, when ten subjects made maximal and 50% of maximal static exertions in standing postures. Exertions were made in flexion, extension, and left and right lateral bending. Three heavy-lifting tasks also were studied. A biomechanical model was used to predict the forces in the trunk muscles, and the predictions then were compared to the measurements. The abdominal muscles were all active in attempted flexion, while the erector spinae muscles were inactive. In attempted extension, the erectors were maximally active, but considerable activity was present in the abdominal muscles as well. The highest activity levels recorded in the oblique abdominal muscles were in lateral bending. There were high degrees of correlation between the measured muscle activities and predicted muscle tensions for the erector spinae and rectus abdominus muscles, while the correlation coefficients for the oblique abdominal muscles were lower (0.4-0.7). The study indicates that inclusion of antagonistic activity is an important consideration to improve model predictions. The oblique abdominal muscles appear to be more active, in general, than predicted. For the longitudinal trunk muscles, the predictions are excellent throughout. PMID- 2964728 TI - The role of prerotation of the trunk in axial twisting efforts. AB - The myoelectric activity of selected trunk muscles was recorded during the development of controlled isometric axial torques. Muscle activity was measured bilaterally over the erector spinae, the rectus abdominus, the oblique external and the oblique internal abdominal muscles at the L3 level. Subjects first applied graded isometric torque efforts over a 10 second ramp up to maximum voluntary contraction with the trunk in neutral rotation. They then repeated the effort with the trunk twisted to the left and right. The largest electromyographic activities were found in the agonistic oblique muscles, but considerable antagonistic activity was present also. While the activity of the internal oblique and rectus were bilaterally similar in symmetric standing a difference occurred between the two sides when the trunk was twisted to the right or left. Axial prerotation of the trunk by 30 degrees in the direction of torque development marginally decreased the maximal developed torque, whereas prerotation in the opposite direction increased the developed torque. PMID- 2964729 TI - Intradural spinal tumors and spinal stenosis. Report of two cases. PMID- 2964730 TI - The effect on the lumbosacral spine of long spinal fusion for idiopathic scoliosis. A minimum 10-year follow-up. AB - The effect on the lumbosacral spine of long spinal fusion in adolescent idiopathic scoliosis was studied in 22 patients. The average duration of follow up was 12.8 years. Corrections of the coronal curves through instrumentation were satisfactory. However, a kyphosis was produced at the fused segment of the lumbar spine in more than half of the cases. The unfused intervertebral spaces distally were hypermobile in extension, which probably indicated early disc degeneration. The incidences of back pain and radiological degeneration were found to be low. There was no relationship between back symptoms and type of employment, pregnancies or radiological degeneration. Preservation of normal lumbar lordosis during spinal fusions was emphasized. PMID- 2964731 TI - Serial measurements of fibrinolytic activity in acute low back pain and sciatica. AB - A fibrinolytic defect is common in chronic back pain syndromes. Its role in the chronicity of these conditions is not fully understood. To elucidate the possible mechanisms, 11 patients with acute low back pain were studied over 12 months and compared with controls. The patients showed prolongation of the euglobulin lysis time throughout the study; the fibrin plate lysis area was initially normal but became abnormal within 2 weeks. In five patients, the symptoms resolved and the initial fibrinolytic defect improved. In contrast, the fibrinolytic defect remained in six patients with persistent pain. These results suggest that the fibrinolytic defect is secondary to mechanical damage but, if persistent, may become a secondary pathogenic factor associated with the chronicity of some back pain problems. PMID- 2964732 TI - Compression fracture: an unusual cause of back pain in a patient with a stable posterior spinal fusion. PMID- 2964733 TI - Myocardial hypertrophy. PMID- 2964734 TI - Workers' compensation: coverage, benefits, and costs, 1985. AB - The 84.3 million workers protected by workers' compensation laws in 1985 represented 87 percent of all wage and salary workers in that year. Both the amount of benefits paid to workers and the cost of the program to employers rose substantially from 1984 to 1985. Benefit payments totaled $22.5 billion-14.1 percent higher than in 1984 and the largest annual increase since 1978-79. About two-thirds of the payments in 1985 were money payments ($15.1 billion) and the remainder ($7.4 billion) went for medical care for disabled workers. Private insurance companies made nearly three-fifths of these payments and State funds and self-insured employers each paid about one-fifth of the total benefit amount in 1985. For the first time since 1978, the annual growth in employer costs exceeded the growth in workers' benefits, resulting in a slight decrease in the loss ratio for 1984-85. Employer costs were up nearly 17 percent from the previous year, reaching an estimated $29.3 billion. Covered payrolls increased by 7 percent in that same period. Total benefit payments as a percent of payroll also increased noticeably in 1985. PMID- 2964735 TI - Human jaw muscle motor behaviour. I. Motor drive. AB - Jaw muscle motor behaviour, however complicated, has important implications for the every day dental practice. In recent years the understanding of jaw and other skeletal muscle function has increased considerably. Direct recording of primary afferent discharge in conscious human beings and animals during normal function has caused radical changes of the concepts of muscle receptor function. Central pattern generators at segmental levels and suprasegmental programming centres are important mechanisms behind voluntary and automatic movements of different kinds. The most important proprioceptive function is probably to provide reassurance of correct movement pattern, to adjust the central programming to environmental changes and to directly influence slow movements requiring precision. Muscle spindle receptors contribute to mandibular kinesthesia. Muscle spindles are rarely present in jaw opening muscles. Despite this fact an excitatory reflex similar to the stretch reflex but with longer latency has been demonstrated. Further on a reciprocal organisation with antagonist inhibition has been shown to exist between jaw openers and closers. Motor behaviour of jaw and limb muscles thus seem to have many characteristics in common. PMID- 2964736 TI - [Atrial hormone]. PMID- 2964737 TI - Changes in worker's compensation law. PMID- 2964738 TI - Comparative study of Biarison and Voltaren in acute lumbar pain and lumbo ischialgia. PMID- 2964739 TI - Resting myocardial ischemia after intravenous infusion of BM 13.177 (sulotroban), a thromboxane receptor antagonist. PMID- 2964740 TI - A simple method for producing degradation products of fibrinogen by an insoluble derivative of plasmin. PMID- 2964741 TI - Relative sensitivity to double thawing of plasma samples for some tests used in thrombosis research. PMID- 2964742 TI - Effect of anticoagulants on shear-induced platelet alterations. PMID- 2964743 TI - Total and free protein S in nephrotic syndrome. PMID- 2964744 TI - [AIDS in children]. AB - Infection with the human immunodeficiency virus type I in children causes variable clinical symptoms. It can lead to asymptomatic disease as well as to acquired immunodeficiency syndrome. In this review we describe the clinical picture, the immunological abnormalities, the mode of transmission, and the preventive measures in case of HIV-I infection. PMID- 2964745 TI - Evaluation of fibrinolytic therapy by measuring cross-linked fibrin derivatives and plasmin-alpha 2-plasmin inhibitor complex in plasma. AB - Plasma levels of alpha 2-plasmin inhibitor (alpha 2PI), plasmin-alpha 2PI complex and cross-linked fibrin derivatives (XDP) were measured in 8 patients (12 episodes) with thromboembolic disorders on the initial administration of urokinase. In conjunction with a decrease in plasma alpha 2PI activity and antigen, plasmin-alpha 2PI complex increased following urokinase infusion in all cases except one who received a low dose (60,000 units) of urokinase. However, changes in XDP were variable among the patients. Plasma XDP level increased markedly in one, moderately in 4, slightly in one, and remained unchanged in 6 cases (episodes). The increment of plasma XDP correlated (r = 0.804, p = 0.003) with the dose of urokinase administered, but was independent of changes in plasmin-alpha 2PI complex. The plasma XDP elevation was associated with clinical improvement. These results suggest that simultaneous measurements of XDP and plasmin-alpha 2PI complex in plasma would be valuable for the pharmacological or hemostatic assessment of thrombolytic therapy. PMID- 2964746 TI - Effects of in vitro and in vivo anticanine T lymphocyte and anticanine class II specific monoclonal antibodies in the beagle. AB - Two murine anticanine lymphocyte monoclonal antibodies, designated T83 and B1F6, were assessed for (1) in vitro antiepitope activity to circulating lymphocyte subsets, their functional effects on lymphoproliferative assays and binding specificities to diverse cell suspensions and tissue sections; (2) in vivo effects after administration on cells expressing the target epitopes, lymphoproliferation, and allograft survival; and (3) the host immune response to the injected murine immunoglobulins. The monoclonal antibody T83 (IgG3) appeared to be specific for a T cell subset with an up-regulating function on lymphoproliferation. It caused a profound depletion of cells with the appropriate epitope after intravenous administration but it bound to lymphocyte membrane epitopes on some cells in peripheral blood that did not become depleted and putatively caused other modulating effects on lymphocyte function. The monoclonal antibody B1F6 (IgG2a, previously described) was immunologically specific in vitro for cells expressing class II major histocompatibility complex antigens. In the dog, this also consisted of 50% of T lymphocytes. After intravenous administration, there were functional effects similar to those of T83. A modest prolongation of survival of renal allografts was observed when both mAbs were used as the sole immunosuppressive agent. These studies also demonstrated the occurrence of natural canine antimurine IgG antibodies. Administration of either monoclonal antibody was followed by a rapid increase in the concentration of the antimouse antibody(s). We postulate that the presence of canine natural antimouse IgG markedly influenced the biologic effects of in vivo administered monoclonals. PMID- 2964747 TI - In vitro immune responsiveness in rats transfused with ultraviolet irradiated donor-specific blood. PMID- 2964748 TI - Immunosuppression: building on Sandimmune (cyclosporine). PMID- 2964749 TI - Immunomodulation: UV radiation. PMID- 2964750 TI - Regression of recurrent membranoproliferative glomerulonephritis type II in a transplanted kidney after plasmapheresis therapy. PMID- 2964751 TI - [Organization of the archives of case histories and collection of data with regard to late results of treatment]. PMID- 2964752 TI - Interactions between lungworms and gastrointestinal worms in calves. AB - Interactions between gastrointestinal worms (Ostertagia ostertagi, Cooperia oncophora) and lungworms (Dictyocaulus viviparus) in calves were studied by assessing the effect of primary infections with either group of worms on the development of homologous or heterologous challenge infections. Primary infections with lungworms resulted in some degree of resistance to challenge with gastrointestinal worms, but this resistance was lower than that found after homologous infection. Primary infections with gastrointestinal worms did not confer any resistance to challenge with lungworms. On the contrary, an indication was found of some enhancing effect of previous gastrointestinal worm infection on the establishment of lungworms. The highest degree of resistance against lungworm challenge was found where calves have been primarily infected with lungworms. Lungworm infections produced some elevation of serum pepsinogen levels. Gastrointestinal worms evoked a rise in circulating eosinophils, although this rise was smaller and occurred later than in lungworm-infected calves. Under the conditions of the experiment, the effect of 6000 infective lungworm larvae on weight gain was larger than the effect of 100,000 L3 of Ostertagia ostertagi and 100,000 L3 of Cooperia oncophora. PMID- 2964753 TI - Host range mutants of adenovirus type 12 E1 defective for lytic infection, transformation, and oncogenicity. AB - The human adenovirus type 12 (H12) E1A region encodes two early proteins of 266 amino acid residues (266R) and 235R whilst the H12 E1B promoter directs the synthesis of two major proteins of 163R and 482R. To determine the functions of E1A and E1B in lytic infection and oncogenic transformation we have isolated and characterized a series of H12 E1 mutants. Mutant H12 hr 700 contains a point mutation in exon 1 that alters a single amino acid common to both the 266 and 235R proteins. This mutant synthesized reduced levels of E1 and structural proteins at delayed times in HEK cells, transformed BRK cells, and induced tumors in newborn rats at reduced efficiency compared to wild-type virus. The mutation in H12 in 600 truncates the 266R protein in its unique sequences but this mutant synthesized the 235R, E1B, and structural proteins at delayed times in HEK cells. H12 in 600 was nontransforming but induced rare tumors in newborn rats. A third E1A mutant H12 in 601 synthesized no E1A proteins, reduced levels of E1B and structural proteins at delayed times in lytic infections, and was not a transforming or oncogenic virus. Three E1B mutants were studied in detail. Both H12 hr 703 and H12 in 602 encode N-terminal truncated 482R proteins whereas H12 del 620 encodes an in-frame internally deleted 482R protein. All three synthesized reduced amounts of E1A proteins and the E1B 163R protein, identifying a regulatory function for the 482R protein. None of the E1B mutants could transform and only H12 del 620 could induce rare tumors in newborn rats. These results show that H12 oncogenesis requires the coordinated expression of the E1 proteins. PMID- 2964754 TI - Expression of the cellular p53 protein in cells transformed by adenovirus 12 and viral DNA fragments. AB - Expression of the p53 cellular phosphoprotein was examined in rat cells transformed by adenovirus 12 (Ad12) virions and by fragments of Ad12 DNA. p53 was detected in all the cell lines examined. Steady-state levels of p53 were highest in cells transformed by the entire E1 region rather than by E1A alone. Physical association between p53 and the Ad12 E1B 55K protein was not detected. The Ad12 E1B-encoded 55K protein, but not the Ad12 E1B 17K and 19K proteins, appears to participate in regulating p53 protein levels. PMID- 2964756 TI - [Prospects for the use of electronic computer technology in military hospitals]. PMID- 2964755 TI - Functional and antigenic domains of the dengue-2 virus nonstructural glycoprotein NS-1. AB - The gene coding for the nonstructural glycoprotein of dengue-2 virus was cloned, sequenced, and expressed in Escherichia coli. There was about 70% conservation at the amino acid level with dengue serotypes 1 and 4 suggesting an important common function for this protein. Conserved hydrophobic domains were found both before the amino-terminus and at the carboxy-terminus, consistent with transmembrane roles. Evidence for at least partial translocation of NS-1 through the inner membrane of E. coli was found. Also conserved were two signals for N-linked glycosylation located near the middle of NS-1. Various regions of NS-1 were tested for antigenicity with mouse and rabbit polyclonal and mouse monoclonal antibodies. The mouse polyclonal antibodies, made against a crude dengue-infected mouse brain immunogen, reacted most strongly with N-terminal regions of NS-1, whereas, the rabbit antiserum, made against purified NS-1 protein, reacted strongest with C-terminal regions. These findings suggest that immunogen presentation or species differences could be important. Although most of the monoclonals appeared to be unreactive in Western blots with expressed NS-1 proteins, two appeared to react strongly; the region from amino acid (a.a.) 273 to a.a. 346 was required for antibody binding. This region, located adjacent to the two conserved C-terminal hydrophobic domains, is highly charged and contains 5 of the 10 conserved cysteine residues of NS-1. PMID- 2964757 TI - [Biological effects of leakadin]. AB - 57 patients with disseminated tumors of various sites received 10 daily doses of 600 mg/m2 leakadin intravenously. The immunostimulating effect was observed (OKT4/OKT8 normalization). The data on leakadin pharmacokinetics are discussed. PMID- 2964758 TI - [Effect of chalones on changes in the rat larynx induced by DMBA]. AB - The experiments used 144 non-inbred albino male rats aged 2-3 months. They were divided in 6 study groups and one control group. The animals of groups 1-3 received 0.01 ml infusions of 6.5% solution of DMBA into the larynx twice a week for 6 months. In groups 4-5, animals were treated with acetone and dimethylsulfoxide, respectively. Epidermal chalones were instilled in group 2 1 hr after DMBA infusion, and hepatic ones--in group 3. After 7 and 10 months, the animals were sacrificed with large doses of ether, 6 sections were taken from each larynx and stained with hematoxylin--eosin. The statistically treated results showed chalone treatment to inhibit the development of precancerous lesions in larynx induced by DMBA. The greatest inhibitory effect was registered in epidermal chalone treatment. PMID- 2964759 TI - Preventing occupational low-back pain. PMID- 2964760 TI - [Value of laparoscopic examination in the final diagnosis of clinically unclear adnexal tumors]. PMID- 2964761 TI - [Occupational dermatologic aspects of disinfectants and possibilities of skin protection]. PMID- 2964762 TI - [Occupational dermatoses as predominant occupational diseases in public health and social welfare]. PMID- 2964763 TI - [The bovine pericardium for closing abdominal wall defects]. AB - A preparation procedure by making of biologic implants in their biologic texture was investigated in using closures of abdominal wall defects. Defects of the ventral trunk wall of 10 to 20 cm dimension were closed with a collagen implant of the equal size. The morphologic investigation was carried out after 12 respectively 24 months. The kind of the immigrated cells, the revascularisation, and the integration of the implants were criteria for the reutilization. Exclusive fibrocytes with regular cell density in the whole implant were provable after 12 months. The revascularization was demonstrable by numerous vessels and capillaries. The light-histologic findings were unchanged after 24 months, a distinction between implant and surrounding tissue was possible only in an electro-optical way. PMID- 2964764 TI - Immunology of inflammatory bowel disease. PMID- 2964765 TI - [Percutaneous transluminal angioplasty of the internal iliac artery]. AB - Report on the successful therapy of an erectile impotence by percutaneous transluminal angioplasty of the internal iliac artery in a patient with arterial obliteration. PMID- 2964766 TI - [Technic of open decompression, splinting of the small intestine and closure of the abdominal wall in complicated postoperative ileus]. PMID- 2964767 TI - Important factors in the use of the Dacron ligament. PMID- 2964768 TI - [Anterior ligament repair of the knee with Dacron using an arthroscopic approach]. PMID- 2964769 TI - Effects of endotoxins from Bacteroides intermedius and Escherichia coli on human monocytes in vitro. AB - Endotoxins (lipopolysaccharides, LPSs) from the strictly anaerobic bacterium Bacteroides intermedius and from Escherichia coli were compared with respect to effects on human monocytes. Exposure of monocyte monolayer cultures to the structurally different endotoxins caused morphological changes, demonstrated by large rounded cells containing cytoplasmic granules with the B. intermedius LPS, and smaller cells with several cytoplasmic protrusions with the E. coli LPS after 10 days of culture. The B. intermedius LPS was, at some doses, able to induce an increased level of the lysosomal enzyme acid phosphatase and increased C3b receptor-mediated phagocytosis, while other doses gave no effect or weak suppression compared to control cultures. The stimulatory effects were donor dependent. The E. coli LPS had predominantly suppressive effects on acid phosphatase level and C3b phagocytosis. Both endotoxins resulted in enhanced expression of the activation related cell surface antigen, characterized by the mononuclear phagocyte specific monoclonal antibody 1D5. Biphasic dose-response relationships were observed, particularly pronounced with B. intermedius LPS, giving stimulation of enzyme activity with low (1 ng/ml) and high (0.5 microgram/ml) doses, and suppressed enzyme activity with intermediate doses (0.01 0.10 microgram/ml). The results imply that the effects of endotoxins on different aspects of human monocytes vary considerably; they also suggest that the balance between stimulatory and suppressive properties of a particular endotoxin may influence its net effect on mononuclear phagocyte functions. PMID- 2964770 TI - The validity of immunological studies in human immunodeficiency virus infection: a three-year follow-up of 235 homo- or bisexual persons. AB - During a three-year follow-up study of 235 risk-group people living in Finland, we found a slow increase in the proportion of HIV seropositives (from 9.8% to 14.0%) and a slow but continuous advance in symptoms typical of HIV infection among them. The first immunological changes after HIV seroconversion were an increase in Ts-cells and a decline in antigen-induced T-cell functions (proliferation, IL-2 production). A further decline in antigen-induced responses was seen before early clinical symptoms and signs of HIV infection developed. During later phases of HIV infection the decrease of Th-cells and the elevation of serum Beta-2 microglobulin correlated most strongly with the progression of HIV infection. The mitogen-induced lymphocyte functions of the HIV-infected correlated to Th-cell levels, while the antigen-induced proliferation and IL-2 production showed correlation neither to Th-cells nor to Ts-cells. The poor antigen-induced T-cell responses might be caused by abnormally functioning monocytes, known to be infected by HIV. Our results suggest that in HIV infection there is a continuous but slow disease progression, although individual variation may be great. The immunological parameter best predicting the prognosis of an infected person in the early phases of infection was the antigen-induced lymphocyte functions, and later the amount of Th-cells. PMID- 2964771 TI - Course and mode of action of descending system conveying nucleus raphe magnus induced inhibition of flexion reflex in rats. AB - In lightly anaesthetized rats, the latencies and magnitudes of heat-evoked withdrawal reflexes from the hindlimb were measured electromyographically. Low intensity (20-50 microA) stimulation of the nucleus raphe magnus (NRM) resulted in an inhibition of the flexion reflex (commonly referred to as analgesia) as evidenced by increased latency and decreased amplitude. The effect lasted for several minutes after the stimulation was terminated. Following lesions of the dorsolateral funiculus (DLF) at the neural thoracic levels 7-8, baseline latencies were reduced and the effect of the NRM stimulation was abolished. Lesions of the DLF at lumbar level 1 resulted in unaltered baseline latencies with persistence of the inhibitory effect of NRM stimulation. The results of the present experiment show that pathways exerting a tonic inhibition of the withdrawal reflex, and mediating the effect of electrical stimulation from the NRM, descend in the DLF at the thoracic level of the spinal cord. At the lumbar level, there is a shift away from the DLF. The antinociceptive effect of 20 microA NRM stimulation was partly reduced by pretreatment with the serotonin synthesis inhibitor para-chlorophenylalanine (PCPA) or the opiate naloxone (1 mg kg-1 i.v.). In animals pretreated with PCPA, naloxone (1 mg kg-1 i.v.) completely abolished the analgesic effect of the stimulation. Thus, both serotonergic and opioid systems may be implicated in mediating the analgesia. With 50 microA stimulation the same treatment only partly attenuated the NRM-induced analgesia, indicating an additional non-serotonergic and non-opioid mechanism which requires a higher current intensity for its activation. PMID- 2964772 TI - Peridural morphine reduces skin blood flow in the lower limb in man. PMID- 2964773 TI - Down's syndrome and its association with Alzheimer's disease. AB - The purpose of this study was to ascertain the frequency of Alzheimer's dementia in the relatives of Down's probands and to establish whether these two disorders cluster together. Out of 128 families of Down's probands, only 67 responded to the questionnaire. Twelve cases of Alzheimer's dementia were found in the eight families of 67 probands. There was a significant increase in the frequency of presenile dementia, but no significant increase in senile dementia among the first and second degree relatives of these probands. The methodological problems were discussed and future directions suggested. PMID- 2964774 TI - Azotemia caused by renal artery stenosis: treatment by percutaneous angioplasty. AB - Percutaneous angioplasty of the renal artery was performed in 79 patients who had stable or climbing serum creatinine levels greater than 1.7 mg/dl and hemodynamically significant stenosis of the renal artery. Patients who had nonrenal causes of azotemia, nephropathy caused by iodinated contrast material, or serum creatinine levels that were declining while the patients were receiving medical therapy before angioplasty were excluded from the study. Angioplasty resulted in a significant (greater than 20%) decline in the level of serum creatinine (average, 2.7 mg/dl before to 1.7 mg/dl after) in 43% of these patients during an average follow-up period of 16 months. A significant decrease in the level of serum creatinine was seen in 61% of patients with bilateral stenosis, 38% of patients with unilateral stenosis with absent contralateral renal blood flow, and 38% of patients with unilateral stenosis and normal contralateral renal blood flow. Recapture of lost nephron function was least successful in patients whose levels of serum creatinine were greater than 4.0 mg/dl (14%); this included one (11%) of nine patients who were already on hemodialysis. We conclude that angioplasty of the renal artery can play a major role in the treatment of patients who have mild azotemia and bilateral stenosis of the renal artery. It is less successful in treatment of patients who have severe azotemia and those who have unilateral disease. PMID- 2964775 TI - Down's syndrome, duodenal stenosis/annular pancreas, and a stack of coins. PMID- 2964776 TI - Maternal cigarette smoking, Down syndrome in live births, and infant race. AB - Previous studies have suggested that maternal smoking is negatively associated with a Down syndrome live birth. We analyzed the data of the U.S. Perinatal Collaborative Study in a search for racial variation in Down syndrome risk factors. There were 22 cases in 25,346 live births to smoking mothers (4/10,780 blacks, 18/13,320 whites, and 0/1,246 other races) and 42/29,130 live births to nonsmoking mothers (24/14,665 blacks, 14/11,694 whites, and 4/2,771 others). The crude overall rates per 1,000 live births were 0.4 in black smokers and 1.6 in black nonsmokers but 1.4 in white smokers and 1.2 in white non-smokers. Adjusted for maternal age, the summary relative risk for a Down syndrome live birth to a smoking mother was 0.2 in blacks (95% interval 0.1-0.7) but 1.2 in whites (95% interval 0.6-2.5). Stratification on variables associated with socioeconomic status or gestational age at time of entry into the study did not alter the racial difference. A comparison of smokers with those who never smoked revealed essentially the same trends. Among all nonsmokers the ratio of the maternal age adjusted risks for a Down syndrome live birth in whites compared with blacks was 0.7 (95% interval 0.3-1.3), and among all smokers this ratio was 3.6 (95% interval 1.3-9.9). If the results are not attributable to statistical fluctuation or undetected confounding, then differences in the probability of intrauterine survival of the Down syndrome fetus would appear to be one plausible explanation for the difference. PMID- 2964778 TI - Severely handicapped infants with life-threatening conditions: federal intrusions into the decision not to treat. AB - In recent years the federal government has attempted to intervene in certain family-medical decisions to withhold treatment from seriously handicapped newborns with life-threatening conditions. Invoking section 504 of the Rehabilitation Act of 1973, which prohibits discrimination against "otherwise qualified handicapped" individuals, the Reagan Administration promulgated regulations allowing federal government investigations of such decisions. Recently, the U.S. Supreme Court upheld lower court decisions invalidating these "Baby Doe" regulations. The federal government's fall-back position is reflected in the Child Abuse Prevention and Treatment Amendments of 1984, requiring states accepting funds under the Child Abuse Prevention and Treatment Act to establish and maintain procedures to assure that cases of medical neglect of handicapped infants are investigated by the states. Although the primary oversight of parental decision-making has been returned to the states where it has traditionally belonged, the federal government's definition of medical neglect of handicapped infants with life-threatening conditions is an ethically inadequate response to the complex needs of the handicapped child, the family, the medical profession, and society as a whole. After examining the relevance of Kantian, utilitarian, and Rawlsian ethical positions, the author contends that an effective governmental policy, capable of enforcement and acceptance by the public, must utilize the strengths of each philosophy and reflect the pragmatism of American society. PMID- 2964777 TI - On the origin of chromosome anomaly. PMID- 2964779 TI - U.S. Industries v. Director: "claim" versus "condition" in the analysis of workers' compensation cases. AB - Under most workers' compensation statutes, an injury must "arise out of " and "in the course of" employment in order to qualify as a compensable disability. In U.S. Industries v. Director, the Supreme Court held that the Longshoremen's and Harbor Workers' Compensation Act must be strictly construed to avoid transforming the compensation system into a form of social insurance. In U.S. Industries, the Court denied a disability claim based on an arthritic condition which was manifested while the worker was at home in bed. This Note contends that the Supreme Court neglected to consider pertinent medical realities when analyzing the causation question. Thus, the decision undermines the overall rationale behind workers' compensation legislation. Nonetheless, the Note argues that the case does not relax the requirement of adequately scrutinizing the causative elements underlying any reasonable claim for disability benefits. An analysis adequately accommodating both medical and legal facts, instead of relying upon the vagaries of statutory interpretation, is necessary to improve the efficiency and fairness of workers' compensation disability determinations. PMID- 2964780 TI - Natriuresis and atrial natriuretic factor secretion during inappropriate antidiuresis. AB - The mechanisms responsible for the natriuresis encountered in the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) are not fully understood. The present study explores the role of atrial natriuretic factor (ANF). Eight subjects unable to excrete ingested free water normally (three patients with SIADH and five healthy humans after intranasal administration of desmopressin) underwent a standard oral water loading test. Plasma ANF level and urinary sodium excretion increased during water retention, whereas plasma aldosterone value decreased later. The increment of urinary sodium excretion rate was significantly correlated with that of plasma ANF. In two patients with hyponatremia due to SIADH, plasma ANF levels were increased during the hyponatremic phase of their condition and decreased under water restriction. In one of them, marked natriuresis was observed when the plasma ANF level was high. It is concluded that secretion of ANF is acutely and chronically stimulated during water retention in SIADH and that ANF may be in part responsible for the natriuresis encountered in inappropriate antidiuresis. PMID- 2964781 TI - Relationship of penile brachial pressure index to myocardial infarction and cerebrovascular accidents in older men. AB - Vascular disease is a major cause of impotence in patients over the age of 40. In a prospective study of 130 impotent patients followed for 24 to 36 months, patients with a penile brachial pressure index (PBPI) of 0.65 or less had a significantly greater risk of a myocardial infarction or a cerebrovascular accident than patients with higher PBPIs. It is concluded that impotence in association with a low PBPI should be considered an indicator of a future major vascular event. PMID- 2964782 TI - Uterine artery flow velocity waveforms in normal and growth-retarded pregnancies. AB - Uterine artery flow velocity waveforms were measured by continuous-wave Doppler ultrasound in 15 normal pregnant women studied from midgestation until term. Results were analyzed by calculation of the pulsatility index and the systolic/diastolic blood velocity [corrected] ratio. Both indices decreased from 16 to 20 weeks (indicating a lowering of resistance) and thereafter remained stable until term. Resistance was lower when waveforms were recorded directly over the placenta or from the uterine artery close to the placenta. A group of women with severe intrauterine growth retardation were also studied. In preeclampsia, uterine artery resistance was increased in almost all patients. In pregnancies complicated by intrauterine growth retardation of non preeclamptic origin, a wide range of results was obtained. PMID- 2964783 TI - Closure of laparotomy incisions with or without peritoneal suturing and second look laparoscopy. AB - The effect of peritoneal closure after reproductive surgery by Pfannenstiel incisions was studied clinically and by second-look laparoscopy. No difference was found in the length of hospital stay, the incidence of wound complications, and other postoperative complications after abdominal closure with (group peritoneal closure, n = 168) or without peritoneal suturing (group no peritoneal closure, n = 165). The laparoscopic findings of 63 patients in the group with peritoneal closure and 57 patients in the group without closure were compared with those in 150 infertile women with no history of abdominal surgery (control group). No adhesions to the anterior abdominal wall were found in the control group. The incidence of these adhesions was not statistically different between the group with peritoneal closure (22.2%) and the group without peritoneal closure (15.8%). This study suggests that there is no difference in postoperative complications, wound healing, and adhesions to previous laparotomy incisions after laparotomy closure with or without peritoneal suturing. PMID- 2964784 TI - Effect of ovarian steroids on the secretion of immunosuppressive factor(s) from human endometrium. AB - To elucidate the effect of ovarian steroids on the secretion of immunosuppressive factor(s) from endometrium, supernatants from human endometrial cultures with or without the addition of progesterone or estrogen were compared for their effects on mixed lymphocyte reaction and phytohemagglutinin-induced lymphocyte reactivity. The suppressive activities of supernatants from the proliferative endometria were significantly increased by the addition of progesterone into their explant cultures. The addition of both progesterone and estrogen was also effective in increasing the inhibitory degree of proliferative endometria but that of estrogen alone showed no influence. Supernatants from the secretory endometria had higher suppressive activities than those from proliferative ones and showed no change in suppressive activity by the addition of ovarian steroids. The concentrations of progesterone in supernatants were too low to inhibit immune reactivity. These findings suggest that exogenous or endogenous progesterone may indirectly play an important role in the release of immunosuppressive factor(s) from human endometrium. PMID- 2964786 TI - The reliability of the Motor Development Checklist. AB - A study was conducted to determine the interrater and test-retest reliability of the Motor Development Checklist (Doudlah, 1976). Thirty-six subjects with diagnosed multiple disabilities and severe and profound mental retardation were evaluated by two experienced occupational therapists. Reliability was established by means of the intraclass correlation approach, and results indicated that coefficients ranged from .93 to .99. The study results suggest that the Motor Development Checklist, when administered by experienced raters, can provide consistent information regarding the spontaneous motor performance of persons with multiple disabilities. Implications for practice and future research are briefly discussed. PMID- 2964785 TI - Validation of thermal techniques for measurement of pelvic organ blood flows in the nonpregnant sheep: comparison with transit-time ultrasonic and microsphere measurements of blood flow. AB - Data obtained from a thermal system capable of measuring changes in organ temperature as well as tissue thermal clearance in the uterus and vagina have been compared with blood flow measured continuously with a transit-time ultrasound volume-flow sensor placed around the common internal iliac artery and intermittently with radioactive microspheres in the chronically instrumented nonpregnant sheep. Temperature changes in both the uterus and the vagina correlated well with blood flow changes measured by both techniques after intravenous administration of estradiol or norepinephrine. Thermal clearance did not correlate well with blood flow in the vagina or uterus. These methods may have value in the investigation of blood flow patterns in various clinical situations such as the pelvic pain syndrome and early pregnancy. PMID- 2964787 TI - Effects of hypoglycemia and diabetes on fuel metabolism by rat brain microvessels. AB - Glucose and beta-hydroxybutyrate metabolism were compared in isolated cerebral microvessels from chronically diabetic and hypoglycemic rats. As noted previously, glucose oxidation and conversion to lactate are diminished in rats with streptozotocin-induced diabetes. The decrease in glucose metabolism did not result from selective damage to diabetic vessels during isolation, since the ATP level and the ATP/ADP ratio were similar to those of nondiabetic rats, and O2 consumption was increased. In addition, cerebral microvessel oxidation of beta hydroxybutyrate was enhanced by diabetes. By contrast, microvessels from rats made chronically hypoglycemic by insulinoma engrafting 30 days earlier had a more than twofold increase in glucose oxidation and conversion to lactate, whereas their oxidation of beta-hydroxybutyrate was diminished by 50%. Unlike the insulinoma rats, no consistent increase in glucose metabolism was observed in microvessels from rats made hypoglycemic either by acute insulin administration or by a 4-day infusion of insulin. These results indicate that diabetes, and under some circumstances chronic hypoglycemia, markedly alters fuel metabolism in the cerebral microvasculature. PMID- 2964788 TI - Effects of diabetes and starvation on skeletal muscle branched-chain alpha-keto acid dehydrogenase activity. AB - The activation state of branched-chain alpha-keto acid dehydrogenase (BCDH) was studied in rat hindlimb muscles during starvation and insulinopenic diabetes, conditions in which circulating branched-chain amino acids (BCAA) are increased and their oxidation is accelerated. Muscle BCDH is predominantly inactive (phosphorylated) in postabsorptive rats but is activated by increased circulating leucine. Diabetes (streptozotocin-induced and spontaneous BB/W) increased circulating BCAA four- to fivefold and BCDH activity approximately threefold. Insulin treatment caused near normalization of circulating BCAA without correcting BCDH activity. Adrenalectomy of diabetics decreased (without normalizing) circulating BCAA and BCDH activation. Starvation caused mild, progressive increases in circulating BCAA and significant activation of BCDH only after 4 days. Leucine infusion activated BCDH in muscle but the activation by leucine was markedly blunted by diabetes. In isolated perfused hindlimbs (control and diabetic) insulin did not affect BCDH significantly; perfusion with leucine activated BCDH, and this response appeared blunted in diabetics. Activation of muscle BCDH may contribute to increased BCAA catabolism in diabetes; the blunted activation response to hyperleucinemia may spare BCAA and contribute to their persistent elevation in plasma. PMID- 2964789 TI - ANF and exocrine pancreas: ultrastructural autoradiographic localization in acinar cells. AB - Atrial natriuretic factor (ANF) binding sites have been recently demonstrated to be present in exocrine pancreas by an in vitro autoradiographic approach. An autoradiographic study was carried out to identify the exocrine cells containing ANF binding sites and to monitor the fate of 125I-labeled ANF in acinar cells after removal of pancreas at specific time intervals (1-30 min) after intravenous administration. At the light microscopic level, silver grains were found over acinar and centroacinar cells. Concomitant injection of an excess of unlabeled ANF inhibited the binding of labeled peptide by approximately 60%. At the electron microscopic level, the time-course study in acinar cells has revealed that of the cell compartments examined, plasma membrane, Golgi apparatus, mitochondria, and zymogen granules, the nucleus had distinct labeling patterns. Plasma membrane was maximally labeled 1 and 2 min after injection with 125I-ANF. Golgi apparatus was significantly labeled from 2 to 30 min after injection, mitochondria from 1 to 30 min after injection, zymogen granules at 1 and 15 min, and the nucleus only at 30 min. The lysosomal compartment was not labeled during the 30-min observation period. These results suggest that after binding to the plasma membrane, ANF is rapidly internalized and distributed to the intracellular organelles as a function of time. Labeling of the zymogen granules suggests that they may bind ANF and that the atrial peptide may be secreted by acinar cells. The significance of association of radioactivity with mitochondria and nuclei remains to be elucidated but may represent intracellular sites of action of ANF complementary to those on plasma membranes. PMID- 2964790 TI - Altered regulation of intestinal ion transport by enteric nerves in diabetic rats. AB - We compared ion transport parameters in isolated ileal mucosa from diabetic rats (8 wk after streptozotocin injection) and littermate controls under basal conditions and in response to electrical transmural stimulation (TS). Stripped ileal mucosa (submucosal plexus intact) was mounted in Ussing flux chambers modified to include stimulating electrodes on opposite sides of the tissue. Under basal conditions unidirectional fluxes of Na+ and Cl- were decreased across mucosa from diabetic rats compared with controls, whereas net fluxes were not significantly different. TS caused a tetrodotoxin (TTX)-sensitive transient increase in short-circuit current (Isc) that was significantly less in tissue from diabetic than control rats. The muscarinic cholinergic receptor antagonist, atropine, significantly reduced the Isc response to TS in ileum from control but not diabetic rats. In addition, the noncholinergic component of the response was smaller. The muscarinic agonist, Urecholine chloride (bethanechol chloride), caused an increase in Isc that was unaffected by pretreatment with TTX and was the same in tissue from control and diabetic rats. Our results suggest that the intestinal abnormalities that occur in diabetes may include a defect in the regulation of ion transport by enteric nerves resulting in an abnormal ability to respond to luminal and other stimuli. PMID- 2964791 TI - Receptors for insulin-like growth factors I and II in rat gastrointestinal epithelium. AB - Distinct receptors for insulin-like growth factors (IGFs) have been characterized in rat intestinal epithelium using 125I-labeled IGF-I and 125I-labeled IGF-II. In jejunal epithelial plasma membranes, IGF-I receptors were observed with a dissociation constant (Kd) of 7.2 nM and a binding capacity of 0.56 pmol/mg protein. Distinct IGF-II receptors were also found with a Kd of 9.5 nM and a binding capacity of 2.61 pmol/mg protein. For IGF-I receptors the following order of affinity was observed: IGF-I greater than IGF-II greater than insulin greater than proinsulin. IGF-II receptors recognize IGF-II with a 20-fold higher affinity than IGF-I and display no cross-reactivity with insulin and proinsulin. Affinity labeling of intestinal membranes also discriminates between the two types of receptors, revealing a radioligand-receptor complex of relative molecular weight (Mr) 130,000 using 125I-IGF-I and 250,000 for 125I-IGF-II under reducing conditions. Separation of proliferative crypt cells from mature villus cells in the small intestine makes it possible to show that a gradient of IGF receptors is present along the crypt-villus axis. 125I-IGF-I and 125I-IGF-II binding is 4.0- and 1.8-fold higher in crypt cells than in villus cells, respectively. Specific 125I-IGF binding is detectable throughout the gastrointestinal tract. The level of IGF binding is similar in stomach, small intestine, and cecum, but higher values are observed in colon. PMID- 2964792 TI - Direct evaluation of the microvascular actions of ANP in juxtamedullary nephrons. AB - The renal vascular actions of atrial natriuretic peptide (ANP) remain incompletely understood. The purpose of this study is to evaluate the effects of ANP on microvascular structures of the normal kidney. The in vitro blood-perfused juxtamedullary nephron technique was utilized to allow visualization of arcuate arteries and afferent and efferent arterioles. Donor rats were pretreated with captopril to eliminate possible interactions between angiotensin II and atriopeptin III (AP III). The effects of topical administration of 3 nM AP III were determined by videometric analysis of vessel inside diameters. Under control conditions, arcuate arterial diameter averaged 83 +/- 14 microns (n = 7), afferent arteriolar diameter was 20 +/- 4 microns (n = 7), and efferent arteriolar diameter was 16 +/- 2 microns (n = 7). During superfusion with AP III, arcuate arteries and afferent arterioles dilated 73 +/- 9 and 23 +/- 5%, respectively. Both returned to their control values when AP III was removed from the superfusate. Further experiments on arcuate arteries (n = 5) revealed that 0.3 nM AP III also vasodilated these vessels (26 +/- 9%); however, no significant effect was elicited by 0.03 nM AP III. In contrast to the vasodilator influence of AP III on preglomerular vessels, efferent arteriolar diameter was not altered by AP III exposure. These observations reveal that AP III can induce selective preglomerular vasodilation involving arcuate arteries as well as afferent arterioles, while efferent arteriolar diameter is not perceptibly influenced. PMID- 2964793 TI - Pressure overload changes cardiac skinned-fiber mechanics in rats, not in guinea pigs. AB - The mechanical properties of detergent-treated skinned fibers from pressure overloaded rat and guinea pig hearts have been compared with those of sham operated animals. Overload was obtained 4 wk after abdominal aortic stenosis, an intervention that increases ventricular weight by 56% in rats and 57% in guinea pigs. The time constant (T, in ms) for tension recovery after a quick stretch was significantly lower in normal guinea pig than in rat. It was lengthened by the process of overload in both species, but this was much more pronounced in rats where T increases by 84% than in guinea pig where it was only slightly augmented by 14% for a doubling of the heart weight. By contrast the maximum tension obtained at pCa 4.5, the stiffness, and the sensitivity to calcium of the fibers were unmodified by chronic overload. In rat, not in guinea pig, a slight decrease in MgATP sensitivity was also observed, whereas no change in creatine kinase efficiency was seen. These results are interpreted as indicating that the slowing of the turnover rate of cross-bridge cycling explains the drop in shortening velocity observed on papillary muscles in rat but not in guinea pig; a species in which membrane modifications must be predominant in the process of adaptation. PMID- 2964794 TI - Renal response to atrial natriuretic factor is modulated by intrarenal angiotensin II. AB - The present study in anesthetized dogs (n = 8) was designed to test the hypothesis that intrarenal angiotensin II (ANG II) attenuates the increase in sodium excretion in response to atrial natriuretic factor (ANF). To test this hypothesis, renal hemodynamic and excretory responses to systemically administered ANF (0.3 micrograms.kg-1.min-1) were assessed in the presence of ANG II infusion into the left kidney (ANG II K) at a nonpressor dose (1.5 ng.kg-1.min 1) and with an infusion of saline into the right kidney, the latter which served as control (CK). During ANF infusion, absolute increases in urinary sodium excretion (delta + 160.8 +/- 44.7 vs. delta + 369.4 +/- 56.9 mu eq/min, P less than 0.005) and fractional sodium excretion (delta + 2.55 +/- 0.62 vs. delta + 4.26 +/- 0.82%, P less than 0.03) were markedly attenuated in the ANG II K compared with CK. Glomerular filtration rate increased only in the CK. Urine osmolality decreased in both the ANG II K and CK. These studies demonstrate an attenuated natriuresis to ANF in the presence of intrarenally infused ANG II, which is associated with a blunted increase in glomerular filtration rate. These studies support the hypothesis that the renal hemodynamic and excretory responses to ANF are modulated by intrarenal ANG II. PMID- 2964795 TI - Adverse cutaneous reactions to antidepressants. AB - The authors review the literature on adverse cutaneous reactions to antidepressant medications. The prevalence of rashes ranges from approximately 2% to 4% but is higher for certain antidepressants such as maprotiline and carbamazepine. Antidepressant drug reactions result in a variety of cutaneous morphologic patterns, but the majority of eruptions are exanthematous. The patterns of these reactions are similar whether the pathogenesis is mediated by immunologic or nonimmunologic mechanisms. The management of patients with adverse cutaneous reactions to antidepressants is discussed, and various recommendations are given. PMID- 2964796 TI - Tolerance and the etiology of alcoholism: hypothesis and mechanism. PMID- 2964797 TI - Dynamic changes in Sertoli cell processes invading spermatid cytoplasm during mouse spermiogenesis. AB - Studies using thick sections stained by ATPase cytochemistry and scanning electron microscopy were carried out to determine three-dimensional ultrastructural alterations in Sertoli cell processes invading neighboring spermatids during mouse spermiogenesis. Sertoli cell processes start invading spermatid cytoplasm at the acrosomal phase of development and undergo considerable change at the maturation phase of development. At step 14, these processes elongate and begin to branch in the spermatid cytoplasm, and by step 15, they extend in various directions to form a complex of canals that the authors have designated the canal complex. The present observations also clarify that the complicated canal complex undergoes regional modification. At the late stages of maturation, the endoplasmic reticulum has gathered with other cell organelles to form aggregates of endoplasmic reticulum in the vicinity of which invading Sertoli cell processes extensively ramify further into thin tubules that intertwine with each other to form a region of thin tubules. In thin sections, each such region was a complex, consisting of small vesicles and endoplasmic reticulum, and corresponded to what has been defined as a mixed body by Morales and Clermont (Anat. Rec., 203:233-244, 1982). During the course of the formation of the region, the invading Sertoli cell processes are continuous at all times with the cell body of the surrounding Sertoli cell. PMID- 2964798 TI - Clinical experiences using pneumatic massage therapy for edematous limbs over the last 10 years. AB - A pneumatic massage apparatus has been developed for the treatment of peripheral lymphedema. The stagnant lymph and venous blood are displaced toward the heart by this pneumatic massage. It has been applied to more than 650 patients with edematous limbs in the authors' Hadomer clinic over the last ten years. Satisfactory results were obtained, including decrease in swelling, pain, and induration. The authors' clinical experiences and statistical analysis of the patients are reported here. STATISTICAL ANALYSIS: 9 primary (4 males and 5 females) and 646 secondary (28 males and 618 females) patients with lymphatic edema of extremities have been treated. The majority of the patients had postmastectomy edema of the upper extremity. CLINICAL STUDIES: The circumference of the edematous limb was measured before and after pneumatic massage treatment of forty to sixty minutes, daily or every second day. (Formula: see text). The rate of swelling was calculated by the formula described above. Twenty-six cases, treated with pneumatic massage for more than fifteen months, have been investigated. A decrease in the rate of swelling was observed in 14 cases (54%), there was no change in 7 (27%), and an increase was seen in 5 (19%). Electric impedance plethysmographic analysis of peripheral lymph volume and blood flow indicated that pneumatic massage was effective in the treatment of edema of the limbs, and its effectiveness was confirmed also by core temperature measurement of the hand during pneumatic massage. The decreased circumference of the edematous limb was kept constant for a long period of time by daily repeated pneumatic massage and ordinary conservative treatments. In conclusion, most lymphedema is of a secondary type. Pneumatic massage was effective in the treatment of peripheral lymph edema, and the improvement was maintained by repeated daily pneumatic massage and ordinary conservative treatment. Therefore, before it becomes worse, lymphedema of the limb should be treated by use of conservative treatments, including pneumatic massage continued as a long-term schedule. PMID- 2964799 TI - Combined thrombolytic therapy and percutaneous transluminal angioplasty for treatment of complex arterial graft thrombosis--a case report. AB - Combined catheter-directed thrombolytic therapy followed by percutaneous transluminal angioplasty (PTA) was successfully performed for the treatment of a thrombosed complex mesenteric artery bypass graft resulting from an anastomotic stenosis. Restenosis of the graft due to neointimal hyperplasia was subsequently treated by PTA, with resultant long-term patency. This approach is an attractive alternative to surgical thrombectomy and graft revision, especially in patients who are poor surgical risks owing to concurrent medical problems. PMID- 2964800 TI - An unusual case of Budd-Chiari syndrome--a case report. AB - The authors report a rare case of congenital Budd-Chiari syndrome in a twenty eight-year-old male mongoloid. The patient was submitted to azygous-portal disconnection, because of the syndrome of portal hypertension supposedly due to cirrhosis of the liver. He died of hemorrhage of the liver on the third postoperative day. Autopsy revealed a congenital fibrotic obstruction of all suprahepatic veins, with a wide, round ligament containing a functional umbilical vein, which had been routinely ligated during surgery. An extensive review of the literature showed no similar report. The authors speculate that the inadvertent interruption of the round ligament, which until then had served as a pathway for venous draining of the liver, followed by ligation of the anastomoses between the portal and azygous systems, was the factor that triggered the lethal outcome. Thus, this appears to be the first case of congenital Budd-Chiari syndrome predominantly maintained at the expense of the round ligament of the liver, with a patent vascular branch. PMID- 2964801 TI - Characterization of fibroblast colony-forming units in bone marrow from healthy cats. AB - The adherent cell layer of bone marrow from healthy cats was characterized in vitro, and the mean fibroblast colony-forming unit (CFU-F) was determined. The majority (82%) of the cells in the adherent cell layer were spindle-shaped fibroblastic cells. These cells were weakly positive for acid phosphatase activity and negative for alpha-naphthyl butyrate esterase and alkaline phosphatase activities. They did not phagocytose latex beads. The remaining cells (18%) in the adherent cell layer resembled macrophages. They were strongly positive for acid phosphatase and alpha-naphthyl butyrate esterase activities, and they phagocytosed latex beads. The mean CFU-F per 10(6) mononuclear cells in bone marrow from healthy kittens and adult cats was 62 and 65, respectively. The CFU-F assay was linear over a range of 0.25 to 1.25 x 10(6) bone marrow mononuclear cells cultured. Variation in the feline CFU-F assay was similar to that reported for the human CFU-F assay. Bone marrow collections repeated at 1 month intervals (from the same bone) did not affect CFU-F concentration. A difference was not observed between CFU-F cultured from the feline humerus or femur. Bone marrow adherent cells in cats resembled those described for other species. Results of the feline CFU-F assay were consistent and repeatable and were similar to those reported for other species. PMID- 2964802 TI - Early experience with a new collagen-impregnated aortic graft. AB - Standard knitted Dacron prosthetic grafts in the aortic position have performed well in terms of patency and durability, but require preclotting and lose their pliability with use. In an effort to overcome these problems, a collagen impregnated double-velour knitted Dacron graft has been developed. This graft is impervious to blood, does not require preclotting, and maintains its pliability. During a 2 and one half year period, 590 of these grafts have been used at our institution for abdominal aortic replacement in both emergent and elective cases. Indications have included aneurysmal disease (67%), occlusive disease (30%), and failed previous aortic graft (3%). A bifurcated graft has been used in most cases (68%). The patient population has included 449 men and 141 women. Their median age is 68 years, with a range from 15 to 91 years. Seventy nine per cent of the patients experienced no perioperative complications. The remaining twenty one per cent have had a variety of complications, most of them cardiac or pulmonary, but none related directly to the graft. Twenty nine patients died in the perioperative period, for a mortality of 4.9 per cent. Late complications have been infrequent and in no cases directly related to the graft. There have been no graft-related late deaths. It is concluded that, at least in this early experience, there are no problems inherent in the use of this graft. Its pliability and the fact that it does not require preclotting are distinct advantages over previous Dacron grafts. PMID- 2964803 TI - [Management of the critically sick child with cardiopathy. Round table]. PMID- 2964804 TI - [Diseases prevalent in adolescents. Round table]. PMID- 2964805 TI - Treatment of the acquired immunodeficiency syndrome (AIDS) and AIDS-related complex with a regimen of 3'-azido-2',3'-dideoxythymidine (azidothymidine or zidovudine) and acyclovir. A pilot study. AB - On the basis of observation that acyclovir potentiates the in-vitro antiviral activity of 3-azido-2',3'-dideoxythymidine (also known as azidothymidine or zidovudine) against human immunodeficiency virus (HIV), we administered a regimen of azidothymidine and acyclovir to eight patients with the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex. An oral regimen of 100 mg of azidothymidine and 800 mg of acyclovir every 4 hours was in general well tolerated, with the principal toxicity being megaloblastic erythroid changes. The pharmacokinetics of the two drugs were independent of each other. Six patients received the drug combination for at least 10 weeks; all had increased numbers of T4+ lymphocytes (P = 0.028), and two of three assessable patients had reversal of anergy. Two patients tested positive for serum HIV p24 antigen at entry, but became negative with treatment. Data for this small group suggest that this drug combination can be tolerated in patients with severe HIV infections; this study can be used as a basis for larger studies of this drug combination. PMID- 2964806 TI - Immunization policies and vaccine coverage among adults. The risk for missed opportunities. AB - A substantial proportion of vaccine-preventable diseases occur among adults. Each year, there are more than 20,000 influenza-associated deaths during epidemics, approximately 40,000 deaths related to pneumococcal disease, and one to five cases of diphtheria. More than 300,000 hepatitis B infections occur annually, mostly in patients 15 to 29 years old. From 1982 to 1986, 96% of patients with tetanus were age 20 and older. Among young adults, 5% to 20% are susceptible to rubella and measles, and outbreaks occur where these persons congregate. Most adults are not immunized, despite recommendations for vaccines against these diseases. Vigorous efforts are needed to implement strategies to reduce disease incidence, morbidity, and death among adults. PMID- 2964807 TI - Antitumor activity of gallium and lanthanum: role of cation-cell membrane interaction. AB - Using the vital dye inclusion method and the radioisotopic study of Ca2+ transport, we have investigated the involvement of the gallium and lanthanum plasma-membrane interaction in the cytotoxic and antitumor characteristics of both cations. The experimental results support the hypothesis that such an interaction is the principal cause of antitumor activity, either through structural changes of the cell membrane or through inhibition of the ATPase pumps. PMID- 2964808 TI - Effects of high-dose medroxyprogesterone acetate and various other steroid hormones on plasma membrane lipid mobility in CAMA-1 mammary cancer cells. AB - The influence of medroxyprogesterone acetate (MPA) and various other steroid hormones on the lateral diffusion of the fluorescent lipid probe 1-acyl-2-(N-4 nitrobenzo-2-oxa-1.3-diazolyl)-aminocarpropylphos phatidylcholine (NBD-PC) in the plasma membrane of intact mammary cancer cells (CAMA-1 cell line) has been studied by fluorescence recovery after photobleaching technique. Incubation of cells with MPA in serum free medium at ambient temperature for 1 hr led to a significant decrease in the lateral diffusion coefficient of NBD-PC. MPA induced this change over a limited concentration range with 10(-7)-10(-5) mol/l near maximal or maximal effects, and 10(-8) mol/l exhibiting no effect. Exposure of the CAMA-1 cells to 10(-7) mol/l MPA in undiluted serum induced a significant effect following 1.5 hr of treatment with no increase in effectiveness up to 4 hr of incubation. As compared to MPA, the other steroids tested were less effective or ineffective. The influence on lateral lipid mobility diminished as follows: MPA greater than progesterone greater than 5a-DHT approximately 17 beta-estradiol greater than dexamethasone, and roughly seems to parallel their lipid solubility as estimated by partition coefficients in an n-octanol-water system. Any involvement of classical steroid hormone receptors in the mechanism of membrane action could be excluded. Nongenomic steroid effects on the plasma membrane are assumed. As the structure and function of biomembranes are modulated by lipid bilayer fluidity and membranes crucially participate in nearly all aspects of cell biology, it is concluded that direct interactions of MPA with membranes potentially play a role in the antitumor activity of the compound. PMID- 2964809 TI - Oral ciprofloxacin treatment for Salmonella typhimurium infection of normal and immunocompromised mice. AB - Oral treatment of Salmonella typhimurium infection with ciprofloxacin was compared with conventional chemotherapy with ampicillin or chloramphenicol in normal (CFW1) and immunocompromised (C57BL/6) mice. Administration of the antibiotics for 12 days reduced the number of bacteria in livers and the mortality of C57BL/6 mice significantly. Ciprofloxacin was considerably more effective than ampicillin in prolongation of the mean survival time of these mice. Similar to conventional chemotherapeutic agents, ciprofloxacin did not prevent fatal disease in most C57BL/6 mice when the treatment lasted 12 days only. On the other hand, ciprofloxacin cured lethal S. typhimurium illness in immunocompromised mice after long-term oral chemotherapy for 26 days at a dosage of 100 mg/kg twice a day. This was not achieved by either ampicillin or chloramphenicol. In normal mice, 12 days of therapy with ciprofloxacin was sufficient for a significant decrease in both the number of viable bacteria in livers and the mortality of lethally infected mice. The results provide a basis for an alternative antibiotic treatment by the oral route in immunocompromised hosts with systemic infections. PMID- 2964810 TI - In vitro culture system to determine MICs and MBCs of antimicrobial agents against Treponema pallidum subsp. pallidum (Nichols strain). AB - A new procedure for determining the susceptibility of Treponema pallidum subsp. pallidum to antimicrobial agents was developed, utilizing a tissue culture system which promotes the in vitro multiplication of this organism. In the absence of antibiotics, T. pallidum (Nichols virulent strain) multiplied an average of 10 fold when incubated for 7 days in the presence of Sf1Ep cottontail rabbit epithelial cell cultures. Varied concentrations of penicillin G, tetracycline, erythromycin, and spectinomycin were added to triplicate cultures to determine their effects on treponemal multiplication, motility, and virulence. The MIC of each antibiotic was defined as the lowest concentration which prevented treponemal multiplication, whereas the MBC was defined as the lowest concentration which abrogated the ability of the cultured treponemes to multiply and cause lesions in rabbits. The in vitro culture technique provided highly reproducible MICs and (in parentheses) MBCs of each of the antibiotics tested: aqueous penicillin G, 0.0005 (0.0025) microgram/ml; tetracycline, 0.2 (0.5) microgram/ml; erythromycin, 0.005 (0.005) microgram/ml; and spectinomycin, 0.5 (0.5) microgram/ml. The significance of these results in light of the in vivo activities and the previous in vitro evaluations of these antibiotics is discussed. The T. pallidum in vitro cultivation system shows promise as a method for studying the interaction between T. pallidum and antimicrobial agents and for screening new antibiotics for syphilis therapy. PMID- 2964812 TI - [A very strange seizure]. PMID- 2964811 TI - [Effects of high-dose medroxyprogesterone acetate in advanced breast cancer, with special reference to prior endocrine therapy and hormone receptors]. AB - Forty-six patients with advanced breast cancer were treated orally with high-dose medroxyprogesterone acetate (MPA), and a 28% (2 CRs, 11 PRs) response rate was obtained. The patient groups that showed a favorable response to MPA were as follows: patients in a postmenopausal state, with soft tissue or bone metastases, with slow-growing tumors, without prior therapy or with good response to prior endocrine therapy, and with positive estrogen (ER)- and/or progesterone (PgR) receptors. Six of 10 patients who had responded to prior endocrine therapy responded to MPA, while only 2 of 23 who had not responded to prior endocrine therapy showed a good response to MPA. There was a significant difference in response to MPA between patients with positive ER or PgR and those with negative ER or PgR tumors. The ER detected in primary or initially relapsed tumors correlated well with the effects of MPA given as a 2nd-line endocrine therapy for advanced breast cancer. The possible roles of MPA in the treatment of advanced breast cancer were discussed. PMID- 2964813 TI - [Circumstances for the cytogenetic diagnosis of trisomy 21]. PMID- 2964814 TI - Prognostic variability among chronic pain patients: implications for study design, interpretation, and reporting. AB - Chronic pain patients share many characteristics, but there is important prognostic variability among them. By selecting for certain characteristics, different recruitment methods and entry criteria for clinical or research programs may influence the likelihood of success regardless of treatment efficacy. This was demonstrated when subjects (n = 55) were recruited through lay publicity for a clinical trial of therapy for chronic back pain. In comparison to routine pain clinic patients (n=61), subjects in the clinical trial were better educated, were more often employed, had more favorable personality profiles, and were less likely to have had surgery or narcotic use (all p less than 0.004). Pain relief was significantly better for clinical trial subjects, apparently due to baseline prognostic differences rather than uniquely efficacious therapy. We conclude that chronic pain patients vary in prognostically important ways; that recruitment methods and criteria strongly influence these characteristics; and that greater attention to these details is needed when interpreting and reporting clinical research. PMID- 2964815 TI - Clinical and experimental evaluation of aortic polytetrafluoroethylene grafts for aneurysm replacement. AB - The aortic expanded polytetrafluoroethylene bifurcation graft has a unique taper design. This was studied in an in vitro model using hydrogen bubbles and colored dyes to assess its hemodynamic significance compared with a conventional nontapered graft model. Under conditions of nonpulsatile flow the only significant differences were seen with a high percentage of occlusions of one outflow limb. Clinical experience with this new graft for infrarenal aneurysm replacement was accumulated in 163 elective cases and 53 emergency cases. The results with this new graft were compared with those obtained in the same time frame with alternative Dacron grafts. No detrimental problems with the graft have appeared in its use up to 78 months following aneurysm bypass or replacement. PMID- 2964816 TI - Hemodynamics of in situ saphenous vein arterial bypass. AB - Doppler-derived blood flow velocity and limb blood pressure measurements were used to characterize the hemodynamics of 128 in situ saphenous vein arterial bypasses. The magnitude and configuration of the graft velocity waveform was the best predictor of clinical outcome. Successful bypasses had antegrade flow throughout the pulse cycle and a blood flow velocity above 40 cm/s. A low graft blood flow velocity (less than 40 cm/s) was associated with technical error or early graft failure due to poor runoff. The return of normal limb blood pressure correlated with a technically satisfactory bypass but was measured in only 50% of limbs on the first day after surgery. In 28 bypasses with high blood flow velocity (mean +/- SD) in systole (102 +/- 20 cm/s) and diastole (35 +/- 11 cm/s), postoperative limb blood pressure was initially low (mean ankle-brachial pressure index = 0.68) due to restriction of blood flow through small-diameter (less than 4-mm) venous conduits. As revascularization hyperemia abated, diastolic blood flow velocity decreased and limb blood pressure normalized. Despite the high blood flow velocity and pressure gradient associated with flow restrictive venous conduits, limb ischemic symptoms resolved, and graft patency was not decreased. An understanding of graft and limb hemodynamics after in situ bypass grafting is critical when noninvasive diagnostic techniques are used to document technical adequacy and for postoperative surveillance. PMID- 2964817 TI - Doppler ultrasound, laser Doppler, and perfusion fluorometry in bowel ischemia. AB - Improved accuracy and objectivity in the evaluation of intestinal viability has been reported by some investigators using Doppler ultrasound, and more recently laser Doppler velocimetry and perfusion fluorometry. To compare the sensitivity and clinical applicability of these techniques, intestinal viability was evaluated by each method in nine 15- to 50-cm loops of small bowel prepared by division of the mesenteric vasculature in five anesthetized dogs. The sensitivity of Doppler ultrasound was 86%, of laser Doppler flow velocity 85%, of laser Doppler index 94%, and of perfusion fluorometry 95%. Though the sensitivity of Doppler ultrasound is significantly less than that of laser Doppler and perfusion fluorometry, this is not unexpected since the latter two techniques are more quantitative than Doppler ultrasound. Clinically, Doppler ultrasound compares favorably with laser Doppler and perfusion fluorometry, and its low cost and simplicity suggest its adjunctive use in the operative setting. PMID- 2964818 TI - [Incidence of the lumbago syndrome in the medical specialty services]. PMID- 2964819 TI - Immunohistochemical studies of complement receptor (CR1) in cases with normal sinus mucosa and chronic sinusitis. AB - The complement receptor (CR1) in the maxillary sinus mucosa of normal patients and in cases of chronic sinusitis was studied with the peroxidase-antiperoxidase, avidin-biotin peroxidase and immunofluorescent methods. CR1 was localized on the ciliary surface and in the cytoplasm of the covering epithelium in both normal controls and the cases of chronic sinusitis. CR1 tended to be denser in the mucosa of chronic sinusitis than in normal mucosa. CR1-binding capacity was also studied with the immunofluorescent method, using C3b-conjugated zymosan. Although CR1 did not bind to C3b in vivo, it was found to bind to C3b in the normal maxillary mucosa when it was treated with C3b-conjugated zymosan. CR1-binding capacity could not be detected in the mucosa from cases with chronic sinusitis, indicating that CR1 was already bound to activated C3b in these cases. PMID- 2964820 TI - Paradoxical stimulation of both lipocortin and prostaglandin production in human amnion cells by dexamethasone. AB - Glucocorticoids inhibit prostaglandin biosynthesis by inducing the formation of lipocortins. In human amnion cells dexamethasone elicited a concentration dependent increase in prostaglandin production and raised intracellular lipocortin 1 concentrations. Dexamethasone could also potentiate the epidermal growth factor (EGF)-induced stimulation of prostaglandin production. EGF alone or in combination with dexamethasone increased lipocortin 1 formation in amnion cells. Human amnion cells may provide a unique insight into interactions between glucocorticoids, lipocortin and eicosanoid biosynthesis. PMID- 2964821 TI - Isolation of heparan sulfates with antithrombin III affinity and anticoagulant potency from BALB/c 3T3, B16.F10 melanoma, and cutaneous fibrosarcoma cell lines. AB - The heparan sulfates synthesized in vitro by three cell lines were isolated by proteolysis and preparative anion exchange chromatography and purified free of other glycosaminoglycans by selective enzymatic degradation. The isolates from the medium of BALB/c 3T3 fibroblasts, B16.F10 melanoma cells, and a cutaneous fibrosarcoma line, along with that from the detergent-extracted cell layer of the fibroblasts, were affinity-fractionated on columns of matrix-immobilized human antithrombin III. Each heparan sulfate contained subfractions with high affinity for the proteinase inhibitor, ranging from 3-34% of the starting material. The high affinity species possessed measurable anticoagulant activities by a clotting assay (6 to 30 units/mg). Since none of the lines were derived from cell types having any known biological role in vascular homeostasis, we suggest that anticoagulant activity of the glycosaminoglycan is a random property of its primary structure. PMID- 2964822 TI - Inhibitor of anion transport, DIDS, releases Ca2+ from hepatic microsomes. AB - Addition of 4,4'-diisothiocyanostilbene-2, 2'-disulfonic acid (DIDS) to Ca2+ loaded hepatic microsomal vesicles evoked a dose-dependent release of the accumulated Ca2+. Ca2+ uptake was also inhibited. The effects of DIDS do not seem to be due to the inhibitions of either Cl- or proton fluxes. The results indicate that DIDS inhibits Ca2+ uptake and releases Ca2+ by inhibiting the Ca2+-ATPase and the formation of the phosphorylated intermediate of the enzyme, and that it might interact with a specific site on the vesicle which is involved in the translocation of Ca2+ across the microsomal and mitochondrial membranes. PMID- 2964823 TI - Differential regulation of insulin-like growth factor-II receptors in rat hepatocytes and hepatoma cells. AB - This study compares the regulation of IGF-II receptors in three rat hepatoma lines, HTC, H-35 and 5123tc, and primary rat hepatocytes. In all cell types [125I]IGF-II bound solely to a species of approximately 250 kDa. Cell surface IGF II receptors in hepatoma cells had slightly lower affinities (1-2 liters/nmol) than in hepatocytes (4 liters/nmol), but slightly higher IGF-I cross-reactivity (2-4% compared to 1% in hepatocytes). In confluent cultures, the three hepatoma lines expressed 5- to 15-fold more cell-surface receptors per cell than hepatocytes. However, while hepatocyte receptors showed marked inverse density dependence, increasing over 6-fold between dense (3 x 10(5) cells/3.8 cm2) and sparse (0.16 x 10(5) cells/3.8 cm2) cultures, receptors in all hepatoma lines remained at a constant high level regardless of culture density. These distinct regulatory patterns resemble those described for growth-related functions in hepatocytes and hepatoma cells, and are thus consistent with a role for IGF-II receptors in liver cell proliferation. PMID- 2964824 TI - [Construction and properties of the expression vector based on the temperature regulated P'R promoter in phage lambda DNA]. AB - Plasmid expression vector using the temperature-regulated promoter P'R of bacteriophage lambda is described. The vector carries a combination of two regions of lambda cI857indgenome, that contain: 1) gene cI and promoter PR, and 2) gene Q and promoter P'R. Transcription or gene Q is initiated at promoter PR, which is controlled by cI857 repressor. The Q gene product acts as a positive regulator of RNA synthesis from P'R. At 37 degrees C, sufficient amounts of protein Q are synthesised to initiate the expression of the cloned gene from P'R. Inactivation of Q gene (by elimination of a single NcoI site) results in the loss of P'R expression activity in the vector. E. coli beta-galactosidase gene (lacZ) and human leukocyte interferon alpha 2 gene (ifn alpha 2) were cloned into a single EcoRI cleavage site under the control of P'R. These constructs express high levels of beta-galactosidase and interferon alpha 2 in E. coli at 37 degrees C. PMID- 2964825 TI - Beta-thromboglobulin during normal pregnancy, labor, and puerperium. AB - Platelet activation has been implicated in the pathogenesis of several obstetrical conditions such as preeclampsia and intrauterine growth retardation. Plasma beta-thromboglobulin (BTG) concentration is an index of in vivo platelet activation. The purpose of this study was to establish normal values of plasma BTG during pregnancy, labor and puerperium. Plasma BTG concentrations were determined from 121 uncomplicated pregnant women, 20 women in labor, 20 puerperal women, and a control group of 28 healthy non-pregnant volunteers. There was no significant difference in plasma BTG concentrations between non-pregnant and pregnant women. Gestational age, labor, and puerperium did not affect plasma BTG concentrations. PMID- 2964826 TI - Muscle fiber types in the cat middle ear muscles. II. Tensor tympani. AB - Using quantitative histochemical techniques, it was determined that the tensor tympani muscle of the cat consists of three muscle fiber types: type 1, type 2A (staining characteristics similar to the type 1 and type 2A muscle fibers found in the control tibialis anterior muscles), and a third unclassified fiber type (type 3) similar to the 2A fiber type except that it had extremely dense alkaline actomyosin adenosine triphosphatase staining (mean transmittance, type 2A = 33.6%; type 3 = 17.3%), as well as dense staining for periodic acid-Schiff, menadione-linked alpha-glycerolphosphate dehydrogenase, nicotinamide-adenine dinucleotide tetrazolium reductase, and succinic dehydrogenase. The type 1 fiber population was smaller in diameter (mean +/- SD, 14 +/- 4 microns) than the type 2A fiber (mean +/- SD, 21 +/- 5 microns) and the type 3 fiber (mean +/- SD, 22 +/ 6 microns) populations. In all muscles, intrafascicular and extrafascicular fat accumulations were found, with the majority being extrafascicular. Calculations indicate that the tendon occupies approximately 41% of the total muscle volume, while the muscle fibers constitute 59% of the volume. PMID- 2964827 TI - The British Paedodontic Society badge. PMID- 2964828 TI - Blast injuries. AB - Blast injuries cause specific lesions with which the radiologist should be familiar. The mechanism of injury and the pathophysiology of this form of trauma are discussed. The clinical effects as well as the radiologic observations in various organs are presented. Most dramatic effects are observed in the thorax. PMID- 2964829 TI - Skeletal scintigraphy of the wrist in suggested scaphoid fracture. AB - Skeletal scintigraphy was performed in 187 patients with clinical suspicion of a scaphoid fracture but with normal radiographic findings. In 85 patients focally moderately (12 patients) or markedly to intensely (73 patients) increased radionuclide uptake was seen, over either the distal radius or carpal bones. A fracture corresponding to markedly increased uptake was eventually verified in 15 out of 26 patients in the distal radius, in 21 out of 28 in the scaphoid but in only 6 out of 18 in other carpal bones. One fracture of the body and one of the hook of the hamate were found, one fracture of the capitate and three of the triquetrum. Not a single fracture was verified in 10 patients with markedly, often intensely, increased activity over the trapezium/trapezoid. It is concluded that scintigraphy is of value in carpal trauma not only to exclude scaphoid fracture but also to direct the attention to the possibility of other carpal fractures, otherwise usually missed. PMID- 2964830 TI - Intraosseous arteriovenous malformations mimicking malignant disease. AB - Primary (congenital) intraosseous arteriovenous malformations are rare, accounting for less than one per cent of all primary intraosseous lesions. They are quite variable in their gross and microscopic presentation, yet all can be traced to anomalous development of the primitive vascular system. They may be totally asymptomatic, cosmetically disfiguring, painful, or on rare occasions, cause high-output cardiac failure. Surgical treatment is often unrewarding with recurrence not uncommon. Intra-arterial embolization has shown promising results. We present three cases of primary intraosseous arteriovenous malformations which on initial work-up mimicked malignant disease. Arteriography proved diagnostic in all three cases. PMID- 2964831 TI - Wegener's granulomatosis in north India. Radiologic manifestations in eleven patients. AB - The chest radiograph in Wegener's granulomatosis reveals several striking, but not pathognomonic abnormalities. Nine (82%) of eleven patients with Wegener's granulomatosis had abnormal chest radiographs. Infiltrates were seen in all patients and were nodular in seven (77%). Six patients had cavities which were most often unilateral and thick walled. Serial radiographs showed the simultaneous appearance and resolution of infiltrates in 4 of 7 instances. Bilateral pulmonary involvement (67%), pleural effusion (33%), and pericardial effusion (22%) were also seen. Paranasal sinus involvement was seen in 4 of 5 studies. The chest radiograph was interpreted as tuberculosis in eight patients, resulting in delayed diagnosis and incorrect treatment. PMID- 2964832 TI - Computed tomography in the diagnosis and treatment of mediastinal abnormalities in children. AB - Thirty-one children with mediastinal abnormalities--14 malignant lymphomas, 4 other primary malignancies, one metastatic and 12 benign lesions--were examined one or several times using CT, which proved to be effective especially for cysts (5 patients), ductus arteriosus aneurysm (2 patients), and intrathoracic liver (one patient). It also supplied important diagnostic information regarding the extent of disease in malignant thymoma (one patient), in neurinoma (one patient), and in Hodgkin's lymphoma (5 patients). It was found to be useful in the monitoring of treatment of patients with lymphomas, in which a small residue, probably a fibrotic remnant, was invariably seen after completion of chemotherapy and irradiation. It was concluded that when the residue was enlarged, the possibility of relapse and even thymic hyperplasia should be considered. However, if CT was performed under general anaesthesia pseudo-widening of the anterior mediastinum could simulate recurrence. Surgical biopsy was found to be necessary in these cases because fine-needle aspiration biopsy was unsuccessful. PMID- 2964834 TI - Renal parenchymal volume in children. Normal values assessed by ultrasonography. AB - Forty-three children with normal urograms and with kidneys of normal size regarding length, area and parenchymal thickness were examined using a water delay computerized ultrasonographic equipment. Renal parenchymal volumes were calculated and related to body weight. The correlation coefficient was 0.92. The renal parenchymal volume of the left kidney was significantly larger than that of the right kidney (p less than 0.001). For practical purposes they should, however, be assumed to be of equal size, 2.0 +/- 0.3 cm3/kg body weight. PMID- 2964833 TI - Scintigraphic evaluation of traumatic splenic lesions in children. AB - Ninety-eight children with recent blunt abdominal trauma which initially evoked clinical suspicion of splenic injury were examined with colloid scintigraphy of the spleen and the liver using multiple imaging views and with abdominal survey. Nineteen children were, in addition, examined with tomographic scintigraphy. The clinical findings and the course of the disorder were reanalyzed. Scintigraphy indicated splenic injury in 56 children and hepatic injury in another 5 children. The left lateral and the left oblique were the optimum imaging views for detecting splenic ruptures. Tomographic scintigraphy did not improve the diagnostic yield. Abdominal survey failed to indicate almost every second case of splenic rupture and provided no additional information of significance. The clinical review agreed with the scintigraphic diagnosis of splenic lesions but, in addition, it suggested possible splenic lesions in another 10 children with normal scintigraphy. This discrepancy cannot be explained as surgery was not employed; the occurrence of splenic lesions too small to become detectable at scintigraphy or to provoke clinically evident symptoms may be supposed. PMID- 2964835 TI - Intravenous injection of ioxilan, iohexol and diatrizoate. Effects on urine profiles in the rat. AB - Effects of intravenous ioxilan, a new third generation non-ionic contrast medium, diatrizoate, iohexol and saline on urine profiles were compared. Albumin, glucose, sodium, phosphate, and the enzymes NAG, LDH and GGT were followed in 24 normal rats over 7 days. Diatrizoate significantly affected all profile components during the first two hours. Albuminuria was significantly greater after diatrizoate than after iohexol or ioxilan, and excretion of glucose, LDH and GGT was significantly higher than after ioxilan. Both iohexol and ioxilan increased the excretion of albumin, LDH and GGT, while iohexol also significantly increased excretion of glucose and sodium. There was a greater excretion of glucose and GGT after iohexol than after ioxilan. Saline did not induce any changes. At day 7, serum sodium, urea, creatinine, and albumin were normal for all test substances, and kidney histology revealed no difference between the groups of animals. It is thus concluded that both high osmolar ionic and low osmolar non-ionic contrast media may cause temporary glomerular and tubular dysfunction in rats. In this model, the kidney is affected most by diatrizoate, less by iohexol, and least by ioxilan. PMID- 2964836 TI - False negative sonographic finding in emphysematous cholecystitis. AB - Two patients presented with fever and acute abdominal pain. Acute cholecystitis was diagnosed when ultrasound examination showed a double-contour gallbladder wall. Radiography showed gas in the gallbladder wall in both cases, indicating emphysematous cholecystitis. This demonstrated that gas in the gallbladder wall may go undetected by ultrasound at the time when it could still be detected by conventional radiography of the abdomen. Awareness of the value of conventional radiography in these cases has an important diagnostic significance which may affect patient management. PMID- 2964837 TI - Magnetic resonance imaging with maximum contrast between two selected tissues composed of recordings with three repetition times. AB - A method is presented which in a composed image maximizes the image intensity difference between two selected tissues. The composition is made from several magnetic resonance (MR) images from the same slice but with different MR imaging parameters. The method is illustrated with a patient with liver metastases for whom the technique was used to emphasize metastases above normal liver parenchyma. PMID- 2964838 TI - Proton relaxation in vivo, in vitro and cellularity. PMID- 2964840 TI - Effect of interferon on T1 relaxation times of liver metastases from endocrine gastrointestinal tumours. AB - Eight patients with liver metastases from endocrine gastrointestinal tumours were examined with magnetic resonance imaging of the liver before and during treatment with interferon. T1, T2 and tumour size were measured and compared with tumour marker levels and symptomatic improvement or deterioration. Before therapy all tumours showed a long T1 and T2, in comparison to normal liver and fat, and during therapy they all showed a decrease in T1. As no change in liver T1 and fat T1 occurred, the decreased tumour T1 is considered to be a therapy effect. This cannot be fully explained but is possibly due to a reduction in tumour growth rate during interferon treatment. There was no certain correlation between tumour T1 and tumour marker levels or symptomatic changes. PMID- 2964839 TI - Control of hepatic tumor hemorrhage by transcatheter embolization with Ethibloc. AB - Two patients with abdominal hemorrhage from a hepatic tumor underwent transcatheter embolization using Ethibloc. The hemorrhage ceased rapidly in both patients. No complications related to the embolization procedure were observed. One patient is doing well after 4 years. The second patient died of extensive metastatic disease eleven weeks after embolization. Control angiography after 14 months and 2 weeks, respectively, demonstrated persistent occlusion of the embolized hepatic arteries. Ethibloc offers the advantage of permanent vascular occlusion and appears to be a safe and effective embolization material for partial occlusion of the hepatic arteries. PMID- 2964841 TI - Distribution of water, fat, and metals in normal liver and in liver metastases influencing attenuation on computed tomography. AB - The quantity of water, lipid and some metals was measured in autopsy specimens of 8 normal livers, 9 livers with fatty change, and in 12 livers with metastases of various origins. These parameters contribute to the CT number measured in the liver. Water played a major role in demonstration of liver metastases as a low density area on CT. Other contributory factors include iron, magnesium and zinc. Lipid and calcium had no influence in this respect. Heavy accumulation of calcium in a metastatic lesion gives a high-density area on CT. However, even when a metastatic lesion was perceived on CT as a low-density area, the calcium content of the lesion was not always lower than that of the non-tumour region. PMID- 2964842 TI - Ultrasonography in acute gallbladder perforation. AB - The files of patients with acute cholecystitis from two large university hospitals from the years 1978-1985 were employed to find the cases with acute gallbladder perforation for this study. Only those patients (n = 9) were selected for the analysis of sonographic signs of acute gallbladder perforation who had less than 48 hours of symptoms before sonography, and were operated upon within 24 hours of the sonography. Patients (n = 10) with non-complicated acute cholecystitis and identical in regard to the duration of the symptoms and the timing of the sonography and the operation formed a control group. The sonographic findings in patients with gallbladder perforation were pericholecystic fluid collections, free peritoneal fluid, disappearance of the gallbladder wall echoes, focal highly echogenic areas with acoustic shadows in the gallbladder, and an inhomogeneous, generally echo-poor gallbladder wall. PMID- 2964843 TI - Radiologic diagnosis of gastro-oesophageal reflux by means of graded abdominal compression. AB - The aim of the study was to ascertain whether abdominal compression by a standardized technique increases the reliability of the radiologic diagnosis of gastro-oesophageal reflux. Eighty-eight patients were examined by means of a manometric reflux test, 24-hour measurements of pH and endoscopy. The radiologic examination was performed with and without graded abdominal compression, the patient at the same time turning from side to side. When abdominal compression was included the result of the radiologic examination was positive in 31 patients. In the absence of abdominal compression reflux was detected in 11 of these patients. In one patient reflux was detected only without compression. All these 31 patients had one more test positive and in 27 at least 2 more tests were positive. Another 21 patients had reflux disease as indicated by at least 2 of the 3 other tests. The compression technique seems to yield no false positive responses, and proved to be significantly more reliable than examinations without compression. PMID- 2964844 TI - Effect of glucagon-(1-21)-peptide on gastroduodenal motility during upper gastrointestinal examination. Randomized trials. AB - The inhibitory effect of glucagon-(1-21)-peptide on the motility of the stomach and duodenum during radiologic double contrast studies was examined. In a randomized trial of equimolar doses of glucagon and glucagon-(1-21)-peptide on 70 patients, glucagon-(1-21)-peptide showed to have a significantly weaker effect than glucagon. Glucagon caused significant increases in plasma-glucose and plasma insulin, while glucagon-(1-21)-peptide had no such effects. In a subsequent randomized study on 75 patients glucagon-(1-21)-peptide in various doses was tested against placebo. The effect was insignificant from that of placebo at practically all dose levels. No side effects were registered. It is concluded, that glucagon-(1-21)-peptide probably has a certain relaxing effect on the smooth muscle of the stomach and duodenum, but that this effect is too small for practical clinical use. PMID- 2964845 TI - Water-soluble contrast media in radiography of small bowel obstruction. Comparison of ionic and non-ionic contrast media. AB - Fifty patients with possible gastrointestinal obstruction, referred for enteric follow-through examination, were randomized for a double-blind, parallel comparison of the hyperosmolar contrast medium Gastrografin and the low-osmolar Omnipaque. The result was that Omnipaque retained its radiographic density in the small bowel better than Gastrografin. Omnipaque was thus a better alternative than Gastrografin in follow-through examinations of intestinal obstruction. Also, 23 patients out of 28 with small bowel obstruction due to peritoneal adhesions, had spontaneous relief of symptoms during the observation period following contrast medium ingestion with no significant difference between the two media. This indicated that enteric follow-through procedures may have a therapeutic efficacy similar to the treatment of small bowel obstruction using nasogastric suction and gastrointestinal rest. Possible mechanisms for this action of the contrast media are discussed. PMID- 2964846 TI - Direct diverticular inguinal hernia. AB - Nineteen patients with direct diverticular inguinal hernia (DDIH) were examined with herniography and surgically explored. A lump medially in the groin and pain were the prominent clinical manifestations in 18 patients. Physical examination indicated the presence of a DDIH in 6 patients. In 16 patients herniography revealed hernial sacs protruding from the supravesical fossa in 5 and from the medial inguinal fossa in 11. At operation a circumscribed defect was found in the transverse fascia laterally and cranially to the pubic tubercle in all patients. In 16 patients peritoneal hernial sacs were demonstrated at surgery while in 3 only lipomas (fatty hernia) were contained within the defect. In our opinion DDIH is a specific variety of inguinal hernia with a fairly typical clinical presentation and radiographic appearance but probably often overlooked at surgery. PMID- 2964847 TI - Magnetic resonance imaging of renal masses at 0.02 tesla. AB - To determine the usefulness of extremely low field magnetic resonance imaging (MRI) in the evaluation of renal masses 19 patients with 15 tumors (13 renal and 2 renal pelvic carcinomas) and 8 cysts were examined in a 0.02 tesla MRI unit. The findings were compared with results of computed tomography (CT) and ultrasound. Cavography was performed in 6 patients. MRI enabled differentiation between cysts and solid tumors. Tumor extension into the inferior vena cava could be demonstrated in one case and liver metastases in two patients. The image quality was inferior to that reported at higher field strengths and the tumors were more precisely staged by CT and ultrasound. At present, it is unlikely that low field MRI will play any substantial role in the evaluation of renal masses. PMID- 2964848 TI - Computed tomography of the brain in cases with venous vasculitis compared with an age-matched reference group. AB - Patients with a particular, steroid-sensitive headache and often characteristic pathology at orbital phlebography, have been suggested to suffer from venous vasculitis. Fifty such patients were examined with computed tomography (CT) of the brain. The findings were compared with those of an age-matched reference group selected at random to represent normal subjects. The CT examinations were analyzed with respect to size of lateral ventricles and signs of atrophy. In both groups, there was a significant increase of atrophy with age. There was also a significantly higher degree of atrophy in the patient group as compared with the reference group. The findings indicate that the supposedly underlying venous vasculitis is related to early aging and atrophy of the brain. PMID- 2964849 TI - Emergency magnetic resonance examination of patients with spinal cord symptoms. AB - Eighteen consecutive patients with spinal cord symptoms of sudden or relatively sudden onset were examined with magnetic resonance imaging (MRI). The examinations were performed on a 0.3 tesla permanent/resistive imaging system using solenoidal surface coils. MRI revealed epidural tumour in five patients, intramedullary tumour in one, epidural abscess in one, myelitis in two, spontaneous intraspinal epidural haematoma in two, disc herniation in two, traumatic lesions in four and no abnormality in one patient. MRI was found to be capable of non-invasively and painlessly detecting and exactly defining the extent of intraspinal and paraspinal lesions. In some cases the nature of the lesion could be inferred from specific signal characteristics, which is a unique property of MRI. The results strongly suggest that MRI is superior to myelography and other imaging methods and should be regarded as the examination of choice in the emergency examination of patients with spinal cord symptoms. PMID- 2964850 TI - Pulmonary artery sling. Diagnosis by magnetic resonance imaging. AB - Aberrant left-sided pulmonary artery (pulmonary sling) is an uncommon anomaly, which may cause significant respiratory morbidity. The condition should be considered on finding a mass interposed between the trachea and the esophagus on the esophagogram, but diagnosis is usually made with invasive methods. Magnetic resonance imaging (MRI) has been reported as an excellent method for diagnosing cardiovascular anomalies but the findings at MRI in cases of pulmonary artery sling have not been reported previously. In this communication the use of MRI is described for demonstration of pulmonary sling in three patients. PMID- 2964851 TI - Effect of training and experience on radiologic diagnostic performance in midfacial trauma. AB - In a retrospective approach the medical records and radiographs of 618 patients with midfacial trauma were reviewed by radiologists who were classified into four groups according to the length of their training and experience. Initially reported diagnoses were compared with discharge diagnoses, and the impact of training and experience on diagnostic performance was assessed. The same parameters were also investigated in a prospective study. Neither of the investigations revealed any statistically significant improvement in diagnostic performance with increased training and experience. After the basic radiologic education, innate perceptive and cognitive abilities seem to have a larger influence on radiologic diagnostic performance than training and experience. PMID- 2964853 TI - Carpal trauma--a diagnostic problem. PMID- 2964852 TI - Vertebral bone mineral measurement using dual photon absorptiometry and computed tomography. AB - The lumbar spine of 14 cadavers was studied both by 153Gd dual photon absorptiometry (DPA) and quantitative computed tomography (QCT) at 96 and 125 kVp. The intact spine and the individual vertebrae were analyzed. After these measurements the ash content of the vertebral body, the posterior elements, and the transverse processes was determined. The fat content of the vertebral body as well as its volume was also measured. With DPA, the bone mineral content (BMC) determined in situ as well as on excised spine specimens correlated highly with the amount of total vertebral ash (r greater than 0.92, SEE less than 3.2 g). The bone mineral density (BMD, area density) of 3 lumbar vertebrae correlated accurately with the mean ash density of the vertebral body (r greater than 0.81, SEE less than 0.015 g/cm3). The so-called corpus density and central density determinations were less accurate. No difference in accuracy was found between measurements when using 3 mm and 4.5 mm step intervals. Variations in the distribution of mineral between the vertebral body and the posterior elements contribute to the error in predicting vertebral body mineral with DPA. QCT gave a smaller error when a cylindric portion of the vertebral body with a 20 mm diameter was measured compared with one with a 9 mm diameter, when the dual energy technique was used (p less than 0.01). With dual energy QCT a correlation was found between a center segment of 3 vertebrae in the lumbar spine and the mean ash density of the vertebral body of r = 0.92 (SEE = 0.010 g/cm3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964854 TI - The scaphoid fat stripe in the diagnosis of carpal trauma. AB - During a ten-year period scaphoid fracture was suggested in 541 wrists, but in 283 cases could not be confirmed at the initial radiographic examination. These cases were immobilized in plaster of Paris and examined clinically and radiologically 10 to 14 days later. A total of 274 additional radiographic examinations revealed 32 scaphoid fractures not initially recognized. Two fractures were at the retrospective analysis found to have been neglected. In the 249 cases where a fracture was never demonstrated, the immobilization and resulting sick leave were in retrospect considered not to have been indicated. In a retrospective analysis of case records and radiographic material including the configuration of the scaphoid fat stripe (SFS) at the initial radiographic examination the SFS was found to be useful in diagnosing scaphoid fractures. The use of the SFS in differential diagnosis would result in reduction of costs and inconvenience for patients without scaphoid fracture. PMID- 2964856 TI - The microbial colonization of inflamed acne vulgaris lesions. AB - Seventy one papules in the early stages of inflammatory development were isolated from acne vulgaris affected skin and their content of micro-organisms characterized. The progress of lesions prior to excision was monitored by tracing an area of the upper back onto a transparent acetate sheet. This template was used the next day, and in some cases after 3 days, to identify inflamed lesions of less than 1 day and 2-3 days duration. These were biopsied, and pilosebaceous units isolated by micro-dissection, homogenized and microbial populations studied by viable counting and microscopy. Propionibacteria colonised 68% of '1 day' duration lesions and 79% of '3 day' duration lesions; staphylococci colonized 19% and 32% respectively and Pityrosporum spp. (Malassezia furfur) were found in 52% and 68%. Although the prevalence of each microbial group was higher in the more chronic lesions, these differences were not statistically significant. The microbial profile of inflamed lesions was similar both qualitatively and quantitatively to non-inflamed lesions studied previously. These results call into question the role of micro-organisms as the initiators of inflammation in acne vulgaris. PMID- 2964855 TI - The immunolocalization of protein gene product 9.5 using rabbit polyclonal and mouse monoclonal antibodies. AB - In order to assess the potential of protein gene product (PGP) 9.5 as a marker of the nervous and neuroendocrine systems, we examined its immunolocation in human, rat and guinea-pig tissues, using a rabbit polyclonal antiserum and two new mouse monoclonal antisera, I3C4 and 3IA3. Our results demonstrate immunoreactive PGP 9.5 in neurons and nerve fibres at all levels of the central and peripheral nervous system, in many neuroendocrine cells, in part of the renal tubule, in spermatogonia and Leydig cells of the testis, and in ova and in some cells of the pregnant and non-pregnant corpus luteum. In routinely processed tissues, standard immunohistochemical techniques using the polyclonal antibody demonstrated peripheral nerve fibres of all sizes with striking clarity. PMID- 2964858 TI - Acute generalized exanthematous pustular dermatitis induced by diltiazem. PMID- 2964857 TI - Malignant melanoma, benzoyl peroxide and acne: a pilot epidemiological case control investigation. AB - A case-control study utilizing available medical records was carried out to investigate possible links between use of topical benzoyl peroxide for acne, and malignant melanoma. One hundred and fifty nine cases and 213 matched controls were used in the analysis which shows no significant association between the use of benzoyl peroxide and the occurrence of malignant melanoma, nor between the occurrence of acne and malignant melanoma. PMID- 2964859 TI - Acne conglobata in transplant patients treated with isotretinoin. PMID- 2964860 TI - Platelet peroxidase-positive blast cells in transient myeloproliferative disorder with Down's syndrome. AB - Transient myeloproliferative disorder accompanied by Down's syndrome has been characterized as exhibiting self-limiting haematological abnormalities. We studied six patients suffering from this disorder in order to clarify the biological nature of their blast cells. Metaphases of leucocytes stimulated with phytohaemagglutinin (PHA) showed trisomy 21 in all patients except one. The exception was constitutionally trisomy 21 mosaic (46,XY = 89/47,XY,+21 = 11). However, metaphases from the peripheral blood cells (blast cells: 70%) without PHA stimulation showed exclusively trisomy 21. Simultaneous examination for morphology and chromosomal analysis on single colonies revealed that granulocyte macrophage (GM) colonies and an erythroid colony contained only cells with the trisomy 21 karyotype. The blast cells showed positive reactions for platelet peroxidase (PPO) and with monoclonal antibodies against platelet-megakaryocyte antigen (TP 80 and TP 82). In methylcellulose and liquid culture systems, high plating efficiencies were observed, and mainly mature basophils containing histamine developed in the presence of PHA-stimulated leucocyte conditioned medium (PHA-LCM). In vivo, mature neutrophils, basophils, eosinophils or megakaryocytes coexisted with PPO-positive blast cells in the peripheral blood of some patients with this disorder. These findings suggest that transient myeloproliferative disorder is characterized by a proliferation of PPO-positive blast cells with trisomy 21, although some heterogeneity may be seen. PMID- 2964861 TI - A randomized trial of progestogens in the primary treatment of endometrial carcinoma. AB - A randomized trial was arranged in Yorkshire to assess whether progestogens would improve survival as part of the primary treatment of endometrial carcinoma, in conjunction with surgery and radiotherapy. A total of 429 patients was randomized on diagnosis to the progestogen or control group and treated locally according to an agreed protocol. All the patients are now more than 1 year after operation and over half are more than 5 years after operation. The projected overall 5-year survival rate is 76% but contrary to expectation there is no statistically significant difference in survival between the patients treated with progestogens and the control group even after statistical adjustment for known prognostic factors. PMID- 2964862 TI - Activated T lymphocytes in uveitis. AB - Two colour flow cytometry techniques were used to assess the activation stages of peripheral and intraocular T lymphocytes in uveitis. Increased numbers of T lymphocytes bearing the interleukin-2 (IL-2) receptors were found in intraocular fluids or peripheral blood or both of 35/51 patients with uveitis. This increased expression of IL-2 receptors on lymphocytes correlated with increased expression of other early T lymphocyte activation markers, HLA-DR and L-35. Both T helper cells (Leu-3A+) and suppressor cells (Leu 2A+) were activated in vivo. A positive correlation was seen between lymphocyte activation and clinical uveitis activity. In idiopathic uveitis activation of Leu-3A lymphocytes (helper/inducer) was significantly increased, and intraocular activation of the Leu-2A lymphocytes (cytotoxic/suppressor) was significantly decreased. These data show that some patients with idiopathic uveitis have a perturbation of T helper cells. Twenty two of 31 patients with idiopathic uveitis, not associated with systemic disease, had increased peripheral T lymphocyte activation. This finding indicates that in some inflammations believed to be restricted to the eye an abnormal systemic immune activation exists. PMID- 2964863 TI - Primary structure of human placental anticoagulant protein. AB - The primary structure of human placental anticoagulant protein was determined by a combination of amino acid and nucleotide sequencing techniques. The carboxymethylated protein was digested with cyanogen bromide, and the resulting peptides were separated by gel filtration and high-performance liquid chromatography. A total of 239 out of 319 amino acid residues were identified from 7 cyanogen bromide fragments. A full-length cDNA clone encoding placental anticoagulant protein was isolated from a human placenta cDNA library. This clone was 1.6 kilobases long and contained a translation initiation site coding for methionine, 957 nucleotides encoding for the mature protein, a stop codon, a poly(A) recognition site, and a poly(A) tail. Analysis of the tryptic-blocked peptide that originated from the NH2-terminus of the protein showed that the terminal methionine was removed and the adjacent alanine residue was acetylated by posttranslational events. Placental anticoagulant protein is composed of 319 amino acids with acetylalanine as the NH2-terminus and has a high degree of sequence identity with lipocortins I and II. It contains four internal repeats, each including a sequence corresponding to a putative Ca2+-dependent phospholipid binding site. Placental anticoagulant protein is a member of the lipocortin/calpactin family. PMID- 2964864 TI - Protein kinase activity associated with Fc gamma 2a receptor of a murine macrophage like cell line, P388D1. AB - The properties of protein kinase activity associated with Fc receptor specific for IgG2a (Fc gamma 2aR) of a murine macrophage like cell line, P388D1, were investigated. IgG2a-binding protein isolated from the detergent lysate of P388D1 cells by affinity chromatography on IgG-Sepharose was found to contain four distinct proteins of Mr 50,000, 43,000, 37,000, and 17,000, which could be autophosphorylated upon incubation with [gamma-32P]ATP. The autophosphorylation of Fc gamma 2a receptor complex ceased when exogenous phosphate acceptors (casein or histone) were added in the reaction mixture. Casein was found to be a much better phosphate acceptor than histone in this system, as casein incorporated about 32-fold more 32P than histone did. Phosphorylation of casein catalyzed by Fc gamma 2a receptor complex was dependent on casein concentration (maximum phosphate incorporation being at 0.5 mg/mL), increased with time or temperature, was dependent on the concentration of ATP and Mg2+, and was maximum at pH near 8. Casein phosphorylation was significantly inhibited by a high concentration of Mn2+ (greater than 25 mM) or KCl (greater than 100 mM) or by a small amount of heparin (greater than 10 units/mL) and was enhanced about 2-fold by protamine. Casein kinase activity associated with Fc gamma 2a receptor used ATP as substrate with an apparent Km of 2 microM as well as GTP with an apparent Km of 10 microM. Prior heating (60 degrees C for 15 min) or treatment with protease (trypsin or Pronase) of Fc gamma 2a receptor complex almost totally abolished casein kinase activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964865 TI - Initiation codons in mammalian mitochondria: differences in genetic code in the organelle. AB - The bovine mitochondrial gene products ND2 and ND4, components of NADH dehydrogenase, have been purified from a chloroform/methanol extract of mitochondrial membranes, and the human mitochondrial gene products ND2 and cytochrome b have been obtained by similar procedures. They have been identified by comparison of their amino-terminal protein sequences with those predicted from DNA sequences of bovine and human mitochondrial DNA. All of the proteins have methionine as their amino-terminal residue. In bovine ND2, this residue is encoded by the "universal" isoleucine codon AUA, and the sequences of human cytochrome b and bovine ND2 demonstrate that AUA also encodes methionine in the elongation step of mitochondrial protein synthesis. In human ND2, the amino terminal methionine is encoded by AUU, which, as in the "universal" genetic code, is also used as an isoleucine codon in elongation. Thus, AUU has a dual coding function which is dependent upon its context. PMID- 2964867 TI - Catalytic oxidation of 2-aminophenols and ortho hydroxylation of aromatic amines by tyrosinase. AB - The usual substrates of tyrosinase, a copper-containing monooxygenase (EC 1.14.18.1), are monophenols and o-diphenols which are both converted to o quinones. In this paper, we studied the reaction of this enzyme with two new classes of substrates: aromatic amines and o-aminophenols, structural analogues of monophenols and o-diphenols, respectively. They undergo the same catalytic reactions (ortho hydroxylation and oxidation), as documented by product analysis and kinetic studies. In the presence of tyrosinase, arylamines and o-aminophenols are converted to o-quinone imines, which are isolated as quinone anils or phenoxazones. As an example, in the presence of tyrosinase, 2-amino-3 hydroxybenzoic acid (an o-aminophenol) is converted to cinnabarinic acid, a well known phenoxazone, while p-aminotoluene (an aromatic amine) gives rise to the formation of 5-amino-2-methyl-1,4-benzoquinone 1-(4-methylanil). Kinetic studies using an oxygen electrode show that arylamines and the corresponding monophenols exhibit similar Michaelis constants (Km = 0.11-0.49 mM). In contrast, the reaction rates observed for aromatic amines are relatively slow (Kcat = 1-3 min 1) as compared to monophenols (1320-6960 min-1). The enzymatic conversion of arylamines by tyrosinase is different from the typical ones: N-oxidation and ring hydroxylation without further oxidation. This difference originates from the regiospecific hydroxylation (ortho position) and subsequent oxidation of the intermediate o-aminophenol to the corresponding o-quinone imine. Finally, the well-known monooxygenase activity of tyrosinase was also confirmed for the aromatic amine p-aminotoluene, with 18O2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964866 TI - Phosphoenzyme decomposition in dog cardiac sarcoplasmic reticulum Ca2+-ATPase. AB - A five-syringe quench-flow apparatus was used in the transient-state kinetic study of intermediary phosphoenzyme (EP) decomposition in a Triton X-100 purified dog cardiac sarcoplasmic reticulum (SR) Ca2+-ATPase at 20 degrees C. Phosphorylation of the enzyme by ATP in the presence of 100 mM K+ for 116 ms gave 32% ADP-sensitive E1P, 52% ADP- and K+-reactive E2P, and 16% unreactive residual EPr. The EP underwent a monomeric, sequential E1P 17 s-1----E2P 10.5 s-1----E2 + Pi transformation and decomposition in the ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid quenched Ca2+-devoid medium. The calculated rate constant for the total EP (i.e., E1P + E2P) dephosphorylation was 7.8 s-1. The E1P had an affinity for ADP with an apparent Kd congruent to 100 microM. When the EP was formed in the absence of K+ for 116 ms, no appreciable amount of the ADP-sensitive E1P was detected. The EP comprised about 80% ADP- and K+-reactive E2P and 20% residual EPr, suggesting a rapid E1P----E2P transformation. Both the E2P's formed in the presence and absence of K+ decomposed with a rate constant of about 19.5 s-1 in the presence of 80 mM K+ and 2 mM ADP, showing an ADP enhancement of the E2P decomposition. The results demonstrate mechanistic differences in monomeric EP transformation and decomposition between the Triton X 100 purified cardiac SR Ca2+-ATPase and deoxycholate-purified skeletal enzyme [Wang, T. (1986) J. Biol. Chem. 261, 6307-6319]. PMID- 2964868 TI - Characterization and purification of a protease in serum that cleaves proatrial natriuretic factor (ProANF) to its circulating forms. AB - Atrial natriuretic factor (ANF) is synthesized and stored in atrial cardiocytes as a 17-kilodalton (kDa), 126 amino acid polypeptide, proANF, but circulates as smaller, 24 and 28 amino acid peptide fragments of the carboxy terminus of proANF. It has previously been shown that proANF is secreted intact from cultured atrial cardiocytes and can be cleaved by a serum protease to smaller, 3-kDa peptides believed to be the circulating forms. This report describes the purification and characterization of this proANF-cleaving protease from rat serum. The cleavages both of 35S-labeled proANF derived from rat atrial cell cultures, as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/autoradiography, and of a synthetic p-nitroanilide containing substrate were used as assays for the detection of enzyme activity. ProANF-cleaving activity was found in rat serum, with no such activity detectable in rat plasma. Cleavage in serum was not dependent on the presence of platelets or other cellular elements. Complete inhibition of proANF cleavage was obtained with the protease inhibitors benzamidine, leupeptin, phenylmethanesulfonyl fluoride, and diisopropyl fluorophosphate (DFP) but not with aprotinin, soybean trypsin inhibitor, pepstatin, or hirudin. Unlike the vitamin K dependent plasma proteins, the proANF-cleaving protease did not adsorb to barium sulfate. With the sequential application of ion-exchange, hydroxylapatite, lectin affinity, and gel filtration chromatography, a 5000-6000-fold purification of the enzyme from rat serum was achieved. Fractionation of either whole serum or the purified enzyme by gel filtration chromatography revealed a single peak of activity corresponding to a protein with a Stokes radius of 45 A.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964869 TI - Characterization of macrophage adhesion molecule. AB - Macrophage adhesion molecule (MAM), an abundant surface molecule which functions in the adhesion and spreading of guinea pig macrophages on surfaces, is characterized as a heterodimer of the trypsin- and plasmin-sensitive glycopeptide gp160 (MAM-alpha) and the glycopeptide gp93 (MAM-beta). The density of MAM molecules is estimated at 630,000 per macrophage on the basis of quantitative binding of 125I-labeled monoclonal antibody. The glycopeptide subunits display microheterogeneity on isoelectrofocusing; the pI is 5.8-6.3 for gp160 (MAM-alpha) and 6.4-7.0 for gp93 (MAM-beta). A neutrophil gp160, gp93 molecule was shown to be indistinguishable from macrophage MAM on the basis of electrophoresis, isoelectrofocusing, and reactivity with 10 monoclonal antibodies. A related heterodimer of gp93 associated with a larger, antigenically different glycopeptide (gp180,gp93) was identified on circulating lymphocytes. Cumulative properties indicate that MAM is the guinea pig analogue of human Mo1 and mouse Mac-1. PMID- 2964870 TI - Preferential location of bulged guanosine internal to a G.C tract by 1H NMR. AB - A series of double-helical oligodeoxyribonucleotides of sequence corresponding to a frame-shift mutational hot spot in the lambda CI gene, 5'-dGATGGGGCAG, are compared by proton magnetic resonance spectroscopy at 500 MHz of the exchangeable protons. Duplexes containing an extra guanine in a run of two, three, and four G.C base pairs are compared to regular helices of the same sequence and to another sequence containing an isolated bulged G, 5'-dGATGGGCAG.dCTGCGCCATC. The imino proton resonances are assigned by one-dimensional nuclear Overhauser effect spectroscopy. Resonances assigned to the G tract in bulge-containing duplexes are shifted anomalously upfield and are very broad. Imino proton lifetimes are determined by T1 inversion-recovery experiments. The exchange rates of G-tract imino protons in bulged duplexes are rapid compared to those in regular helices and are discussed in terms of the apparent rate of solvent exchange for the isolated G bulge. Delocalization of a bulged guanosine in homopolymeric sequences can explain the observed changes in chemical shift and relaxation times across the entire G.C run, and the chemical shifts can be fit by a simple model of fast exchange between base-paired and unpaired states for the imino protons. This allows us to calculate the relative occupancies of each bulge site. In these sequences, we find the extra base prefers positions internal to the G tract over those at the edge. PMID- 2964871 TI - Effect of mild heat treatment on actin and nucleotide binding of myosin subfragment 1. AB - Chymotryptic subfragment 1 (S-1) prepared from rabbit skeletal myosin has lost its ATPase activity upon incubation at 35 degrees C for 3 h. The loss in ATPase activity was accompanied by the perturbation of the structure of the 50K domain as indicated by a dramatic increase in the tryptic susceptibility of this domain without any change in the susceptibility of the other domains of S-1. The perturbation starts at the C-terminal region of the domain as suggested by the appearance of a 29K intermediate protein band in the tryptic peptide pattern of the heat-treated S-1. The heat-treated molecule essentially retained its actin and polyphosphate binding ability, and the actin binding was still sensitive to the presence of ATP or pyrophosphate. However, as opposed to native S-1, in heat treated S-1 the addition of ATP does not induce an increase in tryptophan fluorescence, and, in the case of the treated species, the fluorescence of 1,N6 ethenoadenosine 5'-diphosphate added to the mixture is quenchable by acrylamide. This latter observation suggests that the binding of the adenine ring of the nucleotide has been altered following the heat treatment. The results indicate that the actin and polyphosphate binding sites of S-1 are distinct and that they are relatively independent of the adenine ring binding site. PMID- 2964872 TI - Characterization of membrane calcium pumps by simultaneous immunoblotting and 32P radiography. AB - Calcium pumps of various plasma membrane, endoplasmic reticulum and sarcoplasmic reticulum preparations were visualized by simultaneous immunoblotting and autoradiography of the 32P-labelled phosphoenzymes. The pump proteins and their fragments produced by a proteolytic pretreatment of the membranes were selectively phosphorylated by [gamma-32P]ATP, separated on an acidic SDS polyacrylamide gel, blotted onto nitrocellulose and reacted with polyclonal antibodies raised against the purified human erythrocyte and rat skeletal muscle sarcoplasmic reticulum calcium pumps, respectively. The immuno-reaction was detected by peroxidase staining, while the phosphoproteins were shown by autoradiography of the same blot. An antibody against the erythrocyte calcium pump, reacting on the blot with the 140 kDa erythrocyte calcium pump and its 80 kDa proteolytic fragment, did not show a cross-reaction with the calcium pump of similar molecular mass in rat synaptosome membranes or with any of the endoplasmic- or sarcoplasmic-type calcium pumps. An anti-sarcoplasmic reticulum calcium pump antibody cross reacted with several sarcoplasmic and endoplasmic calcium pump proteins and their proteolytic fragments but with none of the plasma membrane pumps. This sensitive double-labelling method can be applied to study structural relationships and molecular alterations in various ion pump proteins. PMID- 2964874 TI - Transfer of plasmid-borne tuf mutations to the chromosome as a genetic tool for studying the functioning of EF-TuA and EF-TuB in the E. coli cell. AB - The elongation factor EF-Tu of E. coli is a multifunctional protein that lends itself extremely well to studies concerning structure-function relationships. It is encoded by two genes: tufA and tufB. Mutant species of EF-Tu have been obtained by various genetic manipulations, including site- and segment-directed mutagenesis of tuf genes on a vector. The presence of multiple tuf genes in the cell, both chromosomal and plasmid-borne, hampers the characterization of the mutant EF-Tu. We describe a procedure for transferring plasmid-borne tuf gene mutations to the chromosome. Any mutation engineered by genetic manipulation of tuf genes on a vector can be transferred both to the tufA and the tufB position on the chromosome. The procedure facilitated the functional characterization of some of our recently obtained tuf mutations. Of particular relevance is, that it enabled us for the first time to obtain a mutant tufB on the chromosome, encoding an EF-TuB resistant to kirromycin. It thus became possible to study the consequences for growth of tufA inactivation by insertion of bacteriophage Mu. The preliminary evidence obtained suggests that an EF-TuA, active in polypeptide synthesis, is essential for growth whereas such an EF-TuB is dispensable. PMID- 2964873 TI - Study of pH- and temperature-induced transitions in F-protein (phosphofructokinase) by spectroscopy and microcalorimetry methods. AB - The temperature- and pH-induced transitions in F-protein (phosphofructokinase (ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11] have been studied by means of microcalorimetry and fluorescence and CD spectroscopy. An increase in pH from approx. 6.0 to approx. 8.0 causes a change in the protein state which seems to correspond to a shift of the dimer-tetramer equilibrium in favour of the tetramers. In the absence of phosphate, stability of the protein to temperature- and urea-induced denaturation at pH 6.0 is higher than that at pH 8.0. An addition of 150 mM phosphate results in a pronounced increase in the protein's stability in such a way that the protein becomes more stable at pH 8.0 than at pH 6.0. The shift of the denaturational heat capacity peak induced by the phosphate binding exceeds 25 degrees C at pH 8.0 and 9 degrees C at pH 6.0. PMID- 2964875 TI - Changes in the pattern of androgen formation in vitro by the baboon fetal adrenal gland at mid- and late gestation. AB - In the present study, baboon fetal adrenal cells were obtained at mid- and late gestation and incubated for various intervals to determine simultaneously the effects of length of incubation and stage of development on the pattern of adrenal steroidogenesis. Cells were treated with adrenocorticotropic hormone (ACTH) from 0 to 48 h of incubation, and the concentrations of dehydroepiandrosterone (DHA), DHA-sulfate (DHAS), and androstenedione (delta 4A) were determined in the medium. The secretion of DHA and DHAS by untreated or ACTH treated cells of midgestation increased linearly throughout the 48-h incubation period. In fetal adrenal incubates of late gestation, however, DHA and DHAS concentrations peaked at 3 h and declined thereafter, suggesting that the DHA secreted into the medium was further metabolized by this tissue. Baboon fetal adrenal cells formed similar amounts of DHAS and DHA at midgestation, but greater quantities of DHAS were formed at term. In fetal adrenal incubates of midgestation, DHA concentrations exceeded those of delta 4A by threefold, a relationship which was reversed at late gestation, probably due to the increase in the activity of 3 beta-hydroxy-steroid dehydrogenase with advancing gestation. Because the decline in DHA with time of incubation was also associated with a concomitant decrease in DHAS and no change in delta 4A, it does not appear that formation of these steroids account for the loss of DHA. We conclude that the pattern of androgen metabolism exhibited by fetal adrenal cells obtained at midgestation is different from that at term.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964876 TI - Tissue reaction to fabrics coated with turbostratic carbon: subcutaneous versus vascular implants. AB - A technique allowing the deposition of an adherent thin film of turbostratic, high-density carbon on heat-sensitive polymers was recently developed. The biological response to this biomaterial on yarns and fabrics of the type used in cardiovascular surgery has been studied. Polyester yarns, knitted Dacron sheets and knitted uncrimped Dacron vascular grafts were coated with a thin film (less than 1 micron) of turbostratic carbon using a physical vapour deposition process. Coated and control discs of knitted material, as well as coated and uncoated yarns, were implanted in pairs in the subcutaneous tissue of mice, using for each type of implant two cohorts of 12 animals, with observation periods of 4 and 8 wks respectively. Vascular grafts (8 cm long, 8 mm i.d.) coated with carbon on the luminal side only, were implanted for 12 wks in the infrarenal aortic position in six dogs, and compared to uncoated Dacron grafts of the same dimensions inserted in the same location and for the same duration in the infrarenal aortic position in six control animals. With subcutaneous implants, there was no significant difference in the tissue reaction to either coated or uncoated patches. In contrast, the vascular grafts, all of which were patent upon retrieval, showed a much lower extent of thrombosis on the blood-exposure surface in the case of carbon-coated Dacron, as compared to the luminal surface of control prostheses. The internal capsule (tissue formed between the polymer fabric and the blood interface) was thinner in carbon-coated grafts than in control grafts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964877 TI - Endothelial cell adhesion to vascular prosthetic surfaces. AB - Improving the long term patency of small diameter prosthetic grafts remains an important but elusive objective in vascular surgery. In pursuit of a non thrombogenic surface, we have cultured human-adult endothelial cells, examined their adhesive properties and their ability to colonize the inner surface of a Dacron graft. To examine cell adhesion, endothelial cells were labelled with 111In-oxine and inoculated onto prosthetic wells previously prepared with either cold insoluble globulin (CIG), 1% gelatin, alginate or left untreated as a control. At 100 min, mean percentage adhesion to CIG and gelatin precoated wells was 86.0 +/- 9.9% (+/- SD) and 81.6 +/- 2.9% respectively, whereas alginate at 57.5 +/- 7.5% and the control well at 48.3 +/- 9.9% showed significantly less adhesion. Further experiments examined the adhesion of Indiumoxine labelled endothelial cells to Dacron graft (2 cm X 6 mm ID) placed in an in vitro arterial circuit. An immediate loss of approximately 20% of the cells occurred within the first 30 s, whereafter, a stable population were adherent to the graft material. By 102 min, 73.4 +/- 1.8% of cells remained attached when exposed to tissue culture medium, but only 64.1 +/- 6.9% after exposure to blood. Cultured human adult endothelial cells adhere most effectively to prosthetic surfaces precoated with CIG or gelatin, and remain attached following exposure to shear forces. PMID- 2964878 TI - The nature of the primary photochemical events in rhodopsin and isorhodopsin. AB - The nature of the primary photochemical events in rhodopsin and isorhodopsin is studied by using low temperature actinometry, low temperature absorption spectroscopy, and intermediate neglect of differential overlap including partial single and double configuration interaction (INDO-PSDCI) molecular orbital theory. The principal goal is a better understanding of how the protein binding site influences the energetic, photochemical, and spectroscopic properties of the bound chromophore. Absolute quantum yields for the isorhodopsin (I) to bathorhodopsin (B) phototransformation are assigned at 77 K by using the rhodopsin (R) to bathorhodopsin phototransformation as an internal standard (phi R----B = 0.67). In contrast to rhodopsin photochemistry, isorhodopsin displays a wavelength dependent quantum yield for photochemical generation of bathorhodopsin at 77 K. Measurements at seven wavelengths yielded values ranging from a low of 0.089 +/- 0.021 at 565 nm to a high of 0.168 +/- 0.012 at 440 nm. An analysis of these data based on a variety of kinetic models suggests that the I----B phototransformation encounters a small activation barrier (approximately 0.2 kcal mol-1) associated with the 9-cis----9-trans excited-state torsional-potential surface. The 9-cis retinal chromophore in solution (EPA, 77 K) has the smallest oscillator strength relative to the other isomers: 1.17 (all-trans), 0.98 (9 cis), 1.04 (11-cis), and 1.06 (13-cis). The effect of conformation is quite different for the opsin-bound chromophores. The oscillator strength of the lambda max absorption band of I is observed to be anomalously large (1.11) relative to the lambda max absorption bands of R (0.98) and B (1.07). The wavelength dependent photoisomerization quantum yields and the anomalous oscillator strength associated with isorhodopsin provide important information on the nature of the opsin binding site. Various models of the binding site were tested by using INDO PSDCI molecular orbital theory to predict the oscillator strengths of R, B, and I and to calculate the barriers and energy storage associated with the photochemistry of R and I for each model. Our experimental and theoretical investigation leads to the following conclusions: (a) The counterion (abbreviated as CTN) is not intimately associated with the imine proton in R, B, or I. The counterion lies underneath the plane of the chromophore in R and I, and the primary chromophore-counterion electrostatic interactions involve C15-CTN and C13 CTN. These interactions are responsible for the anomalous oscillator strength of I relative to R and B. (b) The presence of a small activation barrier (~0.2 kcal mol-1) in the 9-cis - 9-trans excited-state surface is associated with the location of the counterion as well as the intrinsic photophysical properties of the 9-cis chromophore. The principal difference between the 1 1-cis -c 1 transphoto reaction surface and the 9-cis - 9-trans photoreaction surface is the lack of effective electrostatic stabilization of distorted 9 = 10 conformations due to incomplete charge polarization. (c) Hydrogen bonding to the imine proton, ifpresent, does not involve the counterion. We conclude that water in the active site, or secondary interactions with the protein (not involving the CTN), are responsible. (d) All photochemical transformations involve one-bond photoisomerizations.This prediction is based on the observation of a very small excited state barrier for the I -- B photoreaction and a negative barrier for the R - B phototransformation, coupled with the theoretical prediction that all two bond photoisomerizations have significant S, barriers while one-bond photoisomerizations have small to negative S, barriers.(e) Rhodopsin is energetically stabilized relative to isorhodopsin due to both electrostatic interactions and conformational distortion, both favoring stabilization of R. The INDO-PSDCI calculations suggest that rhodopsin chromophore-CTN electrostatic interactions provide an enhanced stabilization of -2 kcal mol-1 relative to I. Conformational distortion of the 9-cis chromophore-lysine system accounts for -3 kcal mol-1. (f) Energy storage in bathorhodopsin is-60% conformational distortion and 40% charge separation. Our model predicts that the majority of the chromophore protein conformational distortion energy involves interaction of the C,3(-CH3)=CI4--C,5=N-lysine moiety with nearby (unknown) protein residues. (g) Strong interactions between the counterion and the chromophore in R and I will generate weak, but potentially observable charge-transfer bands in the near infrared. The key predictions are the presence of an observable charge-transfer transition at 859 nm (1 1,640 cm- 1) in I and an analogous, but slightly weaker band at 897 nm (11,150 cm-1) in R. Both transitions involve the transfer of an electron from the counterion into low-lying l theta* molecular orbitals. PMID- 2964880 TI - [Evaluation of the results of arterial reconstructions supplemented by the creation of an arteriovenous anastomosis]. AB - Arterial reconstructions supplemented with arteriovenous anastomosis were studied experimentally. The ischemic changes in the operated dog limb were evaluated by various methods, including electronic diagnostic technique. When the diameter of arteriovenous anastomosis was equal to 40-60 per cent of arterial graft diameter, the best postoperative results were observed. PMID- 2964879 TI - [Endogenous opioid system in the realization of the analgesic effect of alpha tocopherol in reference to algomenorrhea]. AB - Beta-endorphin-like immunoreactivity was studied in 7 patients with algomenorrhea during pain attack and 15 minutes after alpha-tocopherol administration with a therapeutic aim (till the analgetic effect was reached). There was an increase in beta-endorphin-like immunoreactivity after alpha-tocopherol administration. Naloxone administration to 9 patients with algomenorrhea of various etiology resumed the pain. The effect of alpha-tocopherol application for pain relief depended on the pathogenesis of algomenorrhea. At the same time naloxone administration failed to resume the pain in patients, in whom alpha-tocopherol had a strong analgetic effect. It is assumed that the endogenous opioid system participates in alpha-tocopherol effect on pain relief in patients with algomenorrhea. PMID- 2964881 TI - [Effect of adrenaline on the mitosis-inhibiting action of a chalone-containing preparation in Ehrlich ascites tumor]. AB - The effect of adrenaline and Ehrlich ascite carcinoma (EAC) chalone on cell division was studied. It has been established that EAC chalone inhibited cell proliferation. The action of adrenaline was also accompanied by a decrease in mitotic index, but the inhibitory effect of the hormone was weaker than that of chalone, it occurred later and its duration was less. A combined effect of adrenaline and chalone depended on the time interval between the administration of the substances. It has been found that chalone administration 1 h after adrenaline administration prolonged mitotic inhibitory effect by 4 h and its synchronous action on cell division in EAC was weak during the experiment. Combined effect of adrenaline and chalone did not differ from the effect of chalone alone if chalone was administered 3 h after adrenaline administration. PMID- 2964882 TI - Isoproterenol potentiation of methyl mercury effects in vivo on cardiac ATPases and 3H-dopamine uptake. PMID- 2964883 TI - Evaluation of the quantitative analysis of a steroid sulphate using fast atom bombardment and tandem mass spectrometry. AB - Dehydroepiandrosterone sulphate (DHAS) has been quantified in human blood serum by fast atom bombardment (FAB)/tandem mass spectrometry of immunoadsorption extracts. FAB of DHAS yielded abundant ions corresponding to the intact steroid sulphate; these were selected by a double-focusing mass spectrometer prior to collisionally activated decomposition in a quadrupole collision cell and mass analysis by a quadrupole mass filter. [HSO4]- (m/z 97) was the sole prominent daughter ion. For quadrupole mass filter was set to transmit m/z 97 and a narrow range magnet scan yielded a spectrum of parents, including m/z 367 and 369, corresponding to DHAS and the (2H2)-analogue (used as internal standard), respectively. Serum concentrations by this procedure were in good agreement with data obtained by gas chromatographic/mass spectrometric analyses of DHA heptafluorobutyrate, formed by direct derivatization of the steroid sulphate. PMID- 2964884 TI - Effect of the vasodilator peptides calcitonin gene-related peptide and atriopeptin on rabbit microvascular blood flow. Preliminary communication. AB - A neuropeptide known as calcitonin gene-related peptide (CGRP) has been shown to have vasodilatory properties in the rabbit epigastric island flap of almost equivalent potency to prostacyclin. The latter is currently regarded as the most potent vasodilator yet to be isolated. CGRP, with its longer-lasting effect in blood and its ability to overcome noradrenaline-induced vasospasm, may be promising for use during or after ischaemic flap transfer or replantation. Another known vasodilatory peptide, atriopeptin, was not as potent in the microvascular bed of the cutaneous flap as in major blood vessels. PMID- 2964885 TI - A method of phalloplasty using the deep inferior epigastric flap. AB - Penile reconstruction remains a difficult surgical problem. To produce a phallus capable of erection and with a water-tight urethra to the tip has not been solved satisfactorily even by the introduction of free flap transfers. We present a method used in four cases, three transsexuals and one pseudohermaphrodite, in which a phallus was successfully constructed using the deep inferior epigastric flap. PMID- 2964886 TI - Scrotal reconstruction with a rectus abdominis muscle flap. AB - The rectus abdominis muscle flap provides excellent coverage of major scrotal defects. It is a simple, reliable, one-stage procedure which covers the exposed testes and gives a satisfactory aesthetic result and is an ideal choice for covering the contaminated wound. PMID- 2964887 TI - Cardiovascular effects of two new calcium antagonists, PY 108-068 and PN 200-110, in conscious spontaneously hypertensive rats. AB - 1. The acute cardiovascular effects of PY 108-068 and PN 200-110 were studied by means of a computerized analysis of the intra-aortic blood pressure (BP) recorded continuously for 26 h in conscious unrestrained spontaneously hypertensive rats. Both compounds were studied at three doses (50, 100 and 200 micrograms kg-1) and each dose was administered intravenously 5 times (09 h 00 min, 14 h 00 min, 19 h 00 min, 24 h 00 min and 09 h 00 min). Baroreflex sensitivity was measured 1 h following the last injection. 2. The two compounds were found to induce rapid (3 min) and dose-dependent falls in BP. After the first administration, these decreases reached -20% and -35% for systolic BP (SBP) and diastolic BP (DBP) respectively with PY 108-068 (200 micrograms kg-1) and -25% and -45% for SBP and DBP respectively with PN 200-110 (200 micrograms kg-1). 3. The duration of the reduction in BP increased with the dose and was much longer for PN 200-110 (180 min for SBP) than for PY 108-068 (20 min for SBP). 4. A tachycardia was associated with the decrease in BP which did not differ at 200 micrograms kg-1 between PY 108-068 (+ 108 beats min-1) and PN 200-110 (+ 103 beats min-1). Baroreflex sensitivity was not significantly increased by either drug. 5. The 5 repeated injections of PY 108-068 and PN 200-110 evoked similar responses. 6. In conclusion, both compounds exhibited marked hypotensive properties. PN 200-110 appeared to be more suitable for further development since its effects were found to be greater and much longer lasting than those of PY 108-068. PMID- 2964888 TI - Plasma concentrations of atrial natriuretic peptide in hypothyroidism. PMID- 2964889 TI - Acne and psoriasis. PMID- 2964890 TI - Skin reactions and fever with indapamide. PMID- 2964891 TI - Laparoscopic incisions at the lower umbilical verge. PMID- 2964892 TI - Effects of blockade of vasopressin V-1 receptors on post-burn myocardial depression. AB - It has been suggested that post-burn myocardial depression may be due to coronary constriction which results in myocardial ischaemia. It has been demonstrated that the levels of vasopressin, a potent natural constrictor of blood vessels, increase four- to six-fold immediately after thermal trauma. Therefore, this substance could be responsible for post-burn coronary constriction and myocardial depression. This was tested using the dog anaesthetized with sodium pentobarbital receiving a 15 per cent total body surface area full thickness flame burn as the experimental model. Cardiac output was measured by the thermal dilution technique. Arterial blood pressure was sensed by a Stathem P-23 transducer. Cardiac force of contraction was measured by a Walton-Brody strain gauge arch sewn on the left ventricle. The results of this study showed a significant decrease in cardiac output, increase in peripheral resistance and decrease in myocardial force of contraction immediately after thermal trauma in untreated animals. The decrease in cardiac output and increase in peripheral resistance remained for the duration of the experimental observations (3 h). The decrease in force of contraction returned to pre-burn levels 1 h post-burn. Pretreatment of the experimental animals with d(CH2)5 Tyr(Me)AVP (SK&F 100273), a vasopressin V-1 receptor blocking agent, prevented the initial decrease in cardiac output, increase in peripheral resistance and decrease in the force of contraction. A correlation plot of peripheral resistance vs. cardiac force of contraction showed a positive correlation between these two variables in the pretreated animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964893 TI - Effects of experimental diabetes on the vitamin D metabolism of pregnant rats and their fetuses. AB - The effects of streptozotocin-induced diabetes on the vitamin D metabolism of pregnant rats were investigated in mothers and their fetuses, 11 and 14 days after streptozotocin (SZ) injection, i.e., on days 18 and 21 of gestation. In the mothers' plasma, the levels of 25-hydroxycholecalciferol (25OHD) and 1,25 dihydroxycholecalciferol (1,25(OH)2 D) were not different from control levels on day 18, but on day 21, 25OHD had increased, 1,25 (OH)2 D had diminished, and significant hypercalcemia was noted (10.1 +/- 0.27 mg/dl vs. 9.47 +/- 0.19 mg/dl, mean +/- SD). In hyperglycemic fetuses from the diabetic mothers, plasma insulin levels were reduced at day 18 but enhanced at day 21. 25OHD levels were not different from those of the controls at day 18, but were lower at day 21 (2.12 +/ 0.70 ng/g BW, n = 13, vs. 3.75 +/- 1.40 ng/g BW n = 29 controls, means +/- SD). Fetal body levels of 1,25 (OH)2 D were lower than that in the controls at day 18 (16.6 +/- 2.9 pg/g BW, n = 9 x 2, vs. 28.7 +/- 6.3 pg/g BW, n = 7 x 2, mean +/- SD P less than 0.001), but identical to control levels on day 21. The role of fetal or placental enzymes in the regulation of vitamin D metabolism in fetuses is discussed. PMID- 2964895 TI - Dental care, for disabled adults. PMID- 2964894 TI - Primary gastric peripheral T-cell malignant lymphoma with helper/inducer phenotype. First case report with a complete histological ultrastructural and immunochemical study. AB - Primary gastrointestinal T-cell malignant lymphomas (T-ML) are very rare. In this case report we describe a primary gastric tumor with local lymph node involvement. On the basis of histologic, immunohistochemical, and electron microscopic studies, the authors classified this tumor as a pleomorphic T-ML, large cell variant with peripheral helper/inducer T-cell phenotype (Leu1/CD5+, Leu4/CD3+, Leu5/CD2+, Leu9/CD7+, and Leu3/CD4+). The extreme pleomorphism of lymphoma cells, the numerous giant cells, and the presence of tumor nodules with two or three concentric layers were the three striking morphologic features of our case. Tumor cells showed an inconstant but true positive staining with anti LeuM1/CD15 and LeuM3/CD14 antibodies. Vimentin positivity could be related to the presence of intermediate filaments at ultrastructural level. Neuron-specific enolase reactivity was a peculiar but unexplained feature. Furthermore, the positivity of the surface markers Ki-1/CD30, anti-Tac/CD25 and HLA-DR, and the nuclear marker Ki-67 suggested an activation state and a high proliferative activity of the tumor cells. This study emphasizes the usefulness of combined pathologic methods in order to rule other diagnoses such as undifferentiated carcinoma, malignant melanoma, malignant histiocytosis, B-cell lymphoma and interdigitating reticulum cells sarcoma, in view of an extremely polymorph tumor proliferation. This is apparently the first completely documented case report of a primary gastric pleomorphic T-ML of peripheral T-cell origin. PMID- 2964896 TI - Functional properties of proteins coded by three human alpha-interferon genes and a pseudogene. AB - We have characterized the functional properties of four highly purified recombinant human class I alpha-interferon subtypes whose biological activities have not been described previously. We selected biological and biochemical activities that may discriminate between different functions of these molecules. We found that the alpha subtypes could be discriminated only by antiviral-host range specificity and natural killer cell activation. Differences in their antiproliferative activity were cell line dependent. Competitive binding, antiproliferative activity in agar, enhancement of expression of HLA-ABC, elevation of 2'-5'-oligoadenylate synthetase levels and enhancement of phosphorylation of the Mr 69,000 protein kinase did not allow discrimination among the alpha I subtypes on the tested cell lines. PMID- 2964897 TI - Formation of conjugates of 2-fluoro-beta-alanine and bile acids during the metabolism of 5-fluorouracil and 5-fluoro-2-deoxyuridine in the isolated perfused rat liver. AB - We have recently demonstrated that the major biliary metabolite of 5-fluorouracil (FUra) in cancer patients is a conjugate of the FUra catabolite 2-fluoro-beta alanine (FBAL) and cholic acid (D.J. Sweeny, S. Barnes, G. Heggie, and R.B. Diasio. Proc. Natl. Acad. Sci. USA, 84:5439-5443, 1987). This finding prompted us to further examine the metabolism and biliary excretion of clinically relevant concentrations of the fluoropyrimidines FUra and 5-fluoro-2'-deoxyuridine (FdUrd) using the isolated perfused rat liver. During infusion of fluoropyrimidines, rates of appearance of metabolites in bile were similar with both 1 microM FUra and 1 microM FdUrd but were 9-fold higher with 25 microM FUra. Analysis by high performance liquid chromatography demonstrated that unmetabolized fluoropyrimidines and known catabolites (i.e., FBAL) accounted for less than 15% of the total metabolites in bile and that the majority of the biliary metabolites were eluted as three distinct nonpolar compounds. Fast atom bombardment-mass spectrometry demonstrated that these unique metabolites had molecular weights of 497 (peak 1), 497 (peak 2), and 481 (peak 3). These metabolites were hydrolyzed by cholylglycine hydrolase to FBAL and unconjugated bile acids that were identified by gas chromatography-mass spectrometry to be alpha-muricholic acid (peak 1), cholic acid (peak 2), and chenodeoxycholic acid (peak 3). Thus, the major biliary metabolites of FUra and FdUrd were identified as N-(bile acid)-FBAL conjugates. While the N-(bile acid)-FBAL conjugates were the major metabolites in bile, dihydroFUra was the major (greater than 70%) metabolite eliminated into perfusate. In summary, these results demonstrate that FUra and FdUrd undergo similar metabolism in the isolated perfused rat liver and, as was observed in humans, the major biliary fluoropyrimidine metabolites are conjugates of FBAL and bile acids. PMID- 2964898 TI - Potentiation of the gonadotoxicity of Cytoxan in the dog by adjuvant treatment with a luteinizing hormone-releasing hormone agonist. AB - This study evaluates the effect on spermatogenesis of coadministration of Cytoxan (cyclophosphamide) and nafarelin, a luteinizing hormone-releasing hormone agonist. Nafarelin causes complete aspermatogenesis in dogs by interrupting the hypothalamic-pituitary-gonadal axis, which might protect against the testicular cytotoxicity associated with cyclophosphamide. The four treatment groups, each consisting of 2 mature male beagle dogs, were (a) no drug; (b) cyclophosphamide (p.o. 3x weekly for 43 and 48 wk for a total dose of 582 and 709 mg/kg, with dose varying according to weekly hematological profile); (c) nafarelin (2 micrograms/kg s.c. daily for 48 and 52 wk); and (d) cyclophosphamide plus nafarelin [same schedule as above with cyclophosphamide (570 and 698 mg/kg total dose) starting 7 wk after beginning nafarelin]. Plasma testosterone, spermatogenesis, and ejaculate volume were completely suppressed by nafarelin prior to starting cyclophosphamide. By 2 wk after cessation of treatment (posttreatment, PT), plasma testosterone reached normal levels, and at 5 wk PT ejaculates appeared which reached normal volumes 2 to 3 wk later. Normally motile ejaculated spermatozoa were noted at 6 to 8 wk PT in nafarelin-only-treated animals; normal sperm numbers were reached at 14 wk PT. The animals receiving cyclophosphamide plus nafarelin were azoospermic for the entire 65-wk PT period, and at 65 wk PT no germinal cells were found upon evaluation of testicular histology. Sperm numbers in cyclophosphamide-only-treated animals began to rise 10-11 wk PT and reached 150 x 10(6) sperm/ejaculate at approximately 65 wk PT (contemporaneous control dogs had sperm numbers of approximately 300-600 x 10(6)/ejaculate). Spermatogenesis in these cyclophosphamide-only-treated animals was normal in most seminiferous tubules at this time. The addition of nafarelin to cyclophosphamide treatment thus exacerbated the deleterious effects of cyclophosphamide on the testes, suggesting caution for use of such a protocol clinically. PMID- 2964899 TI - Endocytosis and degradation of murine anti-human CD3 monoclonal antibodies by normal and malignant T-lymphocytes. AB - Treatment of lymphoid malignancies with monoclonal antibodies (mAbs) and immunoconjugates is a promising new immunotherapeutic approach. However, few published studies have examined in detail the subcellular fate of antibodies following binding to lymphocyte cell surface antigens. In this study, we have investigated the disposition of monoclonal anti-CD3 antibody 64.1 following binding to normal and malignant T-lymphocytes by using cellular radioimmunoassays and immunoperoxidase and immunogold electron microscopy. Anti-CD3 mAbs were predominantly cleared from the cell membrane at 37 degrees C by receptor-mediated endocytosis, although passive shedding of antibody was also observed. Internalized antibody was sequentially transferred from coated pits to receptosomes and eventually to lysosomes. Intralysosomal degradation appeared to be the ultimate fate of internalized radiolabeled mAbs and was followed by exocytosis of free 125I to the culture medium. Ammonium chloride and monensin were potent inhibitors of lysosomal degradation of 125I-anti-CD3 mAbs and caused intracellular trapping of radiolabeled antibodies. The rapid endocytosis, degradation, and exocytosis of antibody observed in these studies elucidate the mechanism of the improved efficacy of anti-CD3 immunoconjugates when used in conjunction with inhibitors of lysosomal action. PMID- 2964900 TI - Is the brain pain-insensitive? PMID- 2964901 TI - Involvement of serotonin in the central control of opioid secretion. PMID- 2964902 TI - Percutaneous transluminal coronary angioplasty in patients with unstable angina pectoris refractory to medical therapy: long-term clinical and angiographic results. AB - The purpose of this study was to assess the results of percutaneous transluminal coronary angioplasty (PTCA) in 469 consecutive patients with unstable angina pectoris refractory to medical therapy. The primary success rate was 88%, but, since the introduction of the steerable wire system, the success rate has increased to 90%. Mortality was 1%. There were no statistically significant differences in success rates per vessel. Actuarial total 5 year survival was 94%. After 5 years, actuarially, 79% of the patients were free of events (recurrence of angina, residual myocardial infarction, re-PTCA, coronary artery bypass surgery or death). Aortocoronary bypass surgery was performed in 9% of the 469 patients. The angiographic recurrence rate was 28%, but, of the patients who were symptom-free and who had a follow-up angiogram, only 3% had an angiographic recurrence, whereas 98% of the patients who did not have a repeat angiogram were symptom-free. It is concluded that, in a selected group of patients with the clinical syndrome of unstable angina pectoris refractory to medical therapy, the long-term clinical results are good and survival is excellent. PMID- 2964903 TI - Double-wire angioplasty of the right coronary artery bifurcational stenosis. AB - Two cases of distal right coronary artery (RCA) bifurcational stenoses involving ostia of the posterolateral (PLA) and the posterior descending (PDA) branches in patients who underwent successful coronary angioplasty using a double-wire technique are reported. A single guiding catheter and sequential balloon inflations were utilized in one, and two guiding catheters and simultaneous balloon inflations in the other. The indications, techniques, and outcomes are described. PMID- 2964904 TI - Percutaneous brachial catheterization: the hidden hazard of high brachial artery bifurcation. AB - We describe a patient in whom a brachial arterial cutdown done at the same site for percutaneous transluminal angioplasty immediately after uncomplicated percutaneous brachial coronary arteriography revealed the percutaneous sheath in a deeper, smaller, and more lateral artery than the brachial artery, complicating placement of the guiding catheter. This case illustrates the potential hidden hazard of normal brachial artery bifurcation variants, which may be responsible for some of the potential complications encountered in percutaneous brachial artery catheterization techniques. PMID- 2964905 TI - Percutaneous transluminal angioplasty of a surgically obstructed left internal mammary artery graft. AB - We describe a case of an angioplasty of left internal mammary artery. This case had had a recent surgery and was found to have a stenosis at the site of a surgical clip. This was successfully dilated, and the patient became asymptomatic. Follow-up at one year revealed continual improvement. PMID- 2964906 TI - Percutaneous balloon valvuloplasty and coronary angioplasty for the treatment of calcific aortic stenosis and obstructive coronary artery disease in an elderly patient. AB - An 81-year-old man with severe calcific aortic stenosis and coronary artery disease who refused surgical therapy was treated with sequential percutaneous balloon aortic valvuloplasty (PBAV) and percutaneous transluminal angioplasty. Before percutaneous balloon valvuloplasty, the mean aortic gradient was 76 mmHg, and the aortic valve area was .45 cm2. The aortic valve was dilated using 15-mm and 18-mm balloons. The mean gradient decreased to 40 mmHG, and the aortic valve area increased to .62 cm2. Percutaneous transluminal coronary angioplasty (PTCA) was performed 2 weeks later, and an 85% proximal left circumflex stenosis was successfully dilated to 20%. No complications were noted during either procedure. At 6-month follow-up, the patient had returned to normal activities and was asymptomatic. Thus, combined therapy with PBAV and PTCA is technically feasible in selected elderly patients with calcific aortic stenosis and anatomically suitable coronary artery disease. This nonsurgical therapeutic approach may be useful in the treatment of selected patients who refuse or who are deferred from cardiac surgery. PMID- 2964907 TI - Balloon inflation following injection of contrast material through the distal lumen of the USCI balloon catheter. AB - We report on four patients in whom we observed inflation of the balloon following injection of contrast material into the distal lumen during percutaneous transluminal coronary angioplasty. This is a rare technical problem. However, inadvertent balloon inflation may cause transient occlusion of the proximal coronary artery and in one case was associated with acute occlusion of a vessel that had been dilated. Management involves prompt deflation of the balloon. PMID- 2964909 TI - [Changes in the myocardium in rats in hypertrophy induced by aortic ligation]. PMID- 2964908 TI - Diabetes and tolerance in transgenic mice expressing class II MHC molecules in pancreatic beta cells. AB - Insulin-dependent diabetes is caused by the loss of insulin-producing beta cells in pancreatic islets. It has been proposed that aberrant expression of Class II Major Histocompatibility Complex (MHC) molecules on beta cells stimulates an autoimmune attack against beta cell antigens. To test this hypothesis, we generated transgenic mice that express Class II MHC molecules (E alpha d/E beta b, or I-Eb) on beta cells. Diabetes was found in 100% of transgenic progeny from three expressing transgenic mouse lines, but without evidence for lymphocytic infiltrates. Furthermore, T lymphocytes appeared to be tolerant to the transgene I-Eb molecule, despite the absence of expression of I-Eb in the thymus or any other lymphoid tissue. The results suggest that novel expression of Class II MHC molecules on nonlymphoid cells is by itself insufficient to initiate autoimmune responses against tissue-specific antigens. PMID- 2964910 TI - [Histochemical properties of mucopolysaccharides in carcinoma of the stomach]. PMID- 2964911 TI - [Fine needle aspiration cytological diagnosis of breast cancer--a summary of twelve years' experience]. PMID- 2964912 TI - [Fine needle aspiration biopsy of autoimmune thyroiditis and toxic goiter]. PMID- 2964913 TI - [Study on the criteria of fine needle aspiration cytology for the diagnosis and classification of malignant lymphomas]. PMID- 2964914 TI - [Cytodiagnosis of soft tissue tumors by fine needle aspiration]. PMID- 2964915 TI - [CT guided percutaneous fine needle aspiration cytodiagnosis of liver masses]. PMID- 2964916 TI - [Percutaneous transthoracic fine needle aspiration biopsy under X-ray television guidance (an analysis of 100 cases)]. PMID- 2964917 TI - [Analysis of the DNA content of malignant lymphoma cells in fine-needle aspirates]. PMID- 2964918 TI - [Pathological study on the renal toxicity of chelates used in the treatment of heavy metal poisoning]. PMID- 2964919 TI - [Histopathological, autoradiographical and biochemical studies on thallium chondronecrosis and its alleviation by sodium selenite]. PMID- 2964920 TI - [Differentiation and significance of mononuclear cells subsets in human breast cancer stroma]. PMID- 2964921 TI - [Lewis x and sialylated Lewis x-related antigen expression in cancerous and precancerous lesions of colon]. PMID- 2964922 TI - [The investigation on the diagnosis and pathogenesis of Campylobacter-like organisms in gastric mucosal biopsy]. PMID- 2964923 TI - [Immunohistochemical analysis of tumor-draining lymph nodes from patients with gastric cancer]. PMID- 2964924 TI - [Lesions of epitheliosis in the bladder mucosa and its clinical significance]. PMID- 2964925 TI - [Ultrastructural study of 12 cases of malignant mesothelioma]. PMID- 2964926 TI - [Clinicopathological study of 4 cases of juvenile granulosa cell tumor]. PMID- 2964927 TI - [Postnatal development of rat spinal motoneurons with special emphasis on their white matter dendrites: a CT-HPR study]. PMID- 2964928 TI - [Localization of motoneurons innervating the m. ischiocavernosus, m. bulbocavernosus and m. sternocleidomastoideus in the rat with their dendritic distribution revealed by CB-HRP]. PMID- 2964929 TI - [Experimental studies on the effect of anisodamine (654-2) on endotoxic shock]. PMID- 2964930 TI - [Preliminary studies on the physiochemical and biological properties of the tumor anti-migration factor]. PMID- 2964931 TI - [Biological studies and clinical application of the anti-migration factor in tumor diagnosis]. PMID- 2964932 TI - [The effects of electro-acupuncture on exudate volume and TXB2, 6-keto-PGF1 alpha content in the pleural cavity after carrageenin-induced pleurisy in rats]. PMID- 2964933 TI - [Plasma glucose and serum insulin responses in normal adults after ingesting different foodstuffs]. PMID- 2964934 TI - [Ultrastructure of hypoxic HeLa cells]. PMID- 2964935 TI - [A genetic observation on a resistant variation of Bacillus cereus 756 to the new antibiotic 2252]. PMID- 2964936 TI - [Ultrastructural changes in smooth muscle cells in leiomyoma and the myometrium of the human uterus after gossypol treatment]. PMID- 2964938 TI - [The significance of clinical trials]. PMID- 2964937 TI - [Effect of peanut extracts on T-activated red cells]. PMID- 2964939 TI - Gonadotropin-releasing hormone analogs and prostatic cancer. AB - Analogs of GnRH constitute a new category of drugs available for the treatment of advanced prostatic cancer. The efficacy and safety of GnRH analogs in the treatment of this disease is now well established. These compounds represent an important alternative therapy for advanced prostatic cancer patients who do not wish to undergo orchiectomy or for whom DES is not tolerable because of the risk of cardiovascular complications. The advent of the monthly depot form of these drugs will make treatment more convenient and less invasive and will enhance patient compliance. PMID- 2964940 TI - Pathological considerations of reoperative vascular patients. AB - Since vascular disease is always progressive and a perfect vascular prosthesis has yet to be developed, postoperative complications are almost inevitable. In this paper, case histories of those who had to have a second operation or, in other words, re-operated patients, have been examined to ascertain the current problems in vascular surgery. Of 176 vascular reconstructive operations performed between January 1, 1980, and December 31, 1986, 29 re-operations were performed on 19 patients (mean age: 64 years; 15: male). The incidence of late graft failures was 8.4% and, of these, anastomotic aneurysms seemed to be the most serious complication (3.1% incidence rate). Late graft failures included intimal hyperplasia, occurring within two years in five cases, and four cases of progressing atherosclerosis, which appeared three years after the initial operation. In all cases of anastomotic aneurysm, arterial wall failure, possibly combined with the changing of implanted grafts, was considered to be related to the false aneurysmal formation. Knitted Dacron demonstrated susceptibility to atherosclerotic progression, whereas the major fault of polytetrafluoroethylene (PTFE) grafts was the insufficiency of the anastomotic diameter. In conclusion, it was revealed that many factors can provoke late graft failure. Improvement of long-term patency seems to be achieved by a more increased understanding of the pathological meaning of these factors, along with the proper application of medical techniques suited to the causes of vascular obstruction. PMID- 2964941 TI - Effects of interception of snout sensory input on murine neck muscles: an electron microscopic study. AB - To examine whether the interference of the snout sensory input causes neuromuscular dysfunction in the murine neck muscles, the fine structure of the dorsal neck muscles was studied by electron microscopy. The infraorbital nerves were bilaterally transected in the adult ICR mice (one-month-old), and the animals were sacrificed after postoperative periods ranging from six to 18 months. Alpha motor end-plates showed remarkable changes characterized by shrunken synaptic boutons containing abundant lysosomes and/or disappearance of the synaptic boutons. The changes in the muscle spindles were characterized by the sarcolemmal undulation of the intrafusal muscle fibers and the frequent appearance of lysosomes in their sensory terminals. These degenerative changes were observed between nine months and 18 months after neurotomy and intensified with the length of the postoperative period. From the results of the present study, it was concluded that a reflex pathway exists between the snout receptor organs and the dorsal neck muscles. PMID- 2964942 TI - The priming saga: where do we stand now? PMID- 2964943 TI - Respiratory arrest after recovery from anaesthesia supplemented with sufentanil. PMID- 2964944 TI - Hormonal and cardiac effects of converting enzyme inhibition in rat myocardial infarction. AB - To explain how converting enzyme inhibition could improve the prognosis in cardiac insufficiency, the effect of converting enzyme inhibition (CEI) by S9490 3 (Perindopril) treatment for 2 months (treated infarctions, n = 18) on hormonal plasma variables and the quantitative and qualitative changes in myocardium were studied in an experimental model of left ventricular infarction in rats (untreated infarctions, n = 18) and compared to a sham-operated control group (n = 15). Induction of myocardial infarction was associated with a transient decrease in blood pressure. CEI treatment maintained a lower blood pressure throughout the experimental period. Plasma renin concentration was not significantly increased in the untreated infarct group (155.4 +/- 136.7 ng AI/ml/hr) as compared to the sham-operated group (47.6 +/- 15.9 ng AI/ml/hr). Plasma aldosterone did not change in the three experimental groups. The plasma level of immunoreactive atrial natriuretic factor increased in the untreated infarct group (185 +/- 245 pg/ml) as compared with the control group (76 +/- 40 pg/ml) and was normalized by CEI (66 +/- 60 pg/ml). Body weight was slightly decreased in both treated and untreated infarct groups, whereas the heart weight was significantly increased in the untreated group (1,540 +/- 310 mg) and normalized by treatment (1,145 +/- 180 mg) as compared with sham-operated controls (1,071 +/- 80 mg). The combined atria and right ventricular mass was significantly increased in the untreated infarct group (660 +/- 210 mg) and decreased by treatment (443 +/- 106 mg) but was not completely normalized (controls, 343 +/- 40 mg). Left ventricular isomyosin profiles were modified by myocardial infarction as compared with controls: V1 form decreased from 62.4 +/- 9.4% in the sham-operated group to 41.6 +/- 13.4% in the infarct group, and the V3 form increased from 13.0 +/- 4.7% in sham-operated animals to 27.4 +/- 11.8% in untreated infarct animals. CEI treatment partially, but significantly, reversed this modification of the isomyosin profile (V1, 53.0 +/- 14.4%; V3, 17.5 +/- 8.0%). Volume density of collagen was significantly increased in the untreated infarct rats (4.14 +/- 0.81% versus 2.68 +/- 0.49% in controls), and this was reversed by treatment (2.95 +/- 0.66%). Messenger RNA encoding for atrial natriuretic factor, measured by dot blot hybridization, was significantly increased in both the atria and the ventricles in the untreated infarct group, and treatment by CEI partially reversed this increase. Thus, myocardial infarction profoundly modified several variables of peripheral circulation and quantitative and qualitative myocardial protein expression.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2964945 TI - Collagen remodeling of the pressure-overloaded, hypertrophied nonhuman primate myocardium. AB - Cardiac muscle is tethered within a fibrillar collagen matrix that serves to maximize force generation. In the human pressure-overloaded, hypertrophied left ventricle, collagen concentration is known to be increased; however, the structural and biochemical remodeling of collagen and its relation to cell necrosis and myocardial mechanics is less clear. Accordingly, this study was undertaken in a nonhuman primate model of left ventricular hypertrophy caused by gradual onset experimental hypertension. The amount of collagen, its light microscopic features, and proportions of collagen types I, III, and V were determined together with diastolic and systolic mechanics of the intact ventricle during the evolutionary, early, and late phases of established left ventricular hypertrophy (4, 35, and 88 weeks, respectively). In comparison to controls, we found 1) increased collagen at 4 weeks, as well as a greater proportion of type III, in the absence of myocyte necrosis; 2) collagen septae were thick and dense at 35 weeks, while the proportion of types I and III had converted to control; 3) necrosis was evident at 88 weeks, and the structural remodeling and proportion of collagen types I and III reflected the extent of scar formation; and 4) unlike diastolic myocardial stiffness, which was unchanged at 4, 35, or 88 weeks, the systolic stress-strain relation of the myocardium was altered in either a beneficial or detrimental manner in accordance with structural remodeling of collagen and scar formation. Thus, early in left ventricular hypertrophy, reactive fibrosis and collagen remodeling occur in the absence of necrosis while, later on, reparative fibrosis is present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964946 TI - Enhanced sensitivity to hypoxia-induced diastolic dysfunction in pressure overload left ventricular hypertrophy in the rat: role of high-energy phosphate depletion. AB - Isolated buffer-perfused rat hearts with pressure-overload hypertrophy develop a greater decrease in left ventricular (LV) diastolic distensibility and a greater impairment in extent of LV relaxation in response to hypoxia than do normal hearts. Using 31P-NMR spectroscopy, we tested the hypothesis that the enhanced susceptibility of hypertrophied hearts to develop hypoxia-induced diastolic dysfunction is due to an accelerated rate of ATP and/or creatine phosphate depletion. Twelve minutes of hypoxia were imposed on isolated isovolumic (balloon in-left-ventricle) buffer-perfused hearts from 14 rats with pressure-overload hypertrophy (LVH; LV/body wt ratio = 3.43 +/- 17) secondary to hypertension induced by uninephrectomy plus deoxycorticosterone and salt treatment and from 17 age-matched controls (LV/body wt ratio = 2.22 +/- 0.12, p less than 0.001). Coronary artery flow per gram left ventricle was matched in the LVH and control groups during baseline oxygenated conditions and held constant thereafter. Balloon volume was held constant throughout the experiment so that an increase in LV end-diastolic pressure during hypoxia represented a decrease in LV diastolic distensibility. LV systolic pressure was 165 +/- 9 mm Hg in the LVH group compared with 120 +/- 5 mm Hg in the controls during baseline aerobic perfusion (p less than 0.001). LV end-diastolic pressure rose significantly more in response to 12 minutes of hypoxia in the LVH group (12 +/- 1 to 44 +/- 10 mm Hg) than in the controls (12 +/- 1 to 20 +/- 3 mm Hg, p = 0.04). During baseline aerobic conditions, ATP content was the same in the LVH (17.1 +/- 0.5 mumol/g dry LV wt, n = 4) and control (18.8 +/- 0.6 mumol/g dry LV wt, n = 4, p = NS) groups. During hypoxia, ATP declined at the same rate in the LVH and control groups (3.2 +/- 0.5 versus 3.0 +/- 0.5%/min, p = NS) despite the greater rise in end diastolic pressure in the LVH group. Creatine phosphate content during baseline aerobic perfusion was 14% lower in the LVH group compared with controls, but the rate of creatine phosphate depletion during 12 minutes of hypoxia was the same. During hypoxia, intracellular pH declined modestly and to the same degree in both groups. Thus, the greater susceptibility to hypoxia-induced diastolic dysfunction observed in isolated buffer-perfused hypertrophied rat hearts cannot be explained by an initially lower total ATP content or by an accelerated rate of decline of ATP or creatine phosphate.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2964947 TI - Color Doppler flow mapping in patients with coarctation of the aorta: new observations and improved evaluation with color flow diameter and proximal acceleration as predictors of severity. AB - We performed color Doppler flow mapping in 15 patients, 1 week to 17 years old (mean 42 months), with coarctation of the aorta that was confirmed subsequently by angiography and/or surgery. Twelve patients had native coarctation and three had mild recoarctation after surgical repair. Color Doppler flow maps were analyzed with a digital analysis package and a Sony computer system. The diameter in the region of coarctation from the color Doppler flow map (mean = 2.0 +/- 0.8 mm [SD]) correlated well with the coarctation diameter measured at angiography (mean = 1.8 +/- 0.8 mm; r = .83, SEE 0.43 mm) in the 10 patients with native coarctation undergoing angiography, but the coarctation diameter measured by two dimensional echocardiography (3.9 +/- 1.5 mm) was poorly predictive of the angiographic severity (r = .23). Additionally, spatial acceleration was seen in all patients proximal to the coarctation site, with an aliased and accelerating stream narrowing progressively as it proceeded toward the coarctation site, a pattern that is not seen in healthy subjects. Computer analysis of the color Doppler images provided pseudo three-dimensional and digital velocity maps for blue, red, and green (turbulent) flow velocities to allow an enhanced appreciation of the accelerating stream, easily separating this from normal descending aortic aliasing patterns. The narrowing of the acceleration area in the proximal descending aorta (distal/proximal acceleration zone ratio) was also predictive of the angiographic severity of coarctation (r = .83). The distribution of low-level turbulence seen proximally paralleled the distribution of the proximal accelerating stream.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2964948 TI - Clinical outcome 5 years after attempted percutaneous transluminal coronary angioplasty in 427 patients. AB - This study was performed to define the 5 year clinical status of 427 patients who underwent percutaneous transluminal coronary angioplasty (PTCA) in 1981. Their mean age was 54 +/- 10 years (+/- 1 SD). Sixty-one percent had unstable angina, 23% had prior myocardial infarction, 86% had one-vessel disease, and 92% had normal left ventricular function. Sixty-seven percent of patients had left anterior descending artery stenosis. Angiographic success was achieved in 84% of patients. Coronary bypass surgery was required in 9.6% of patients, in 5.9% as an emergency procedure. There were no in-hospital deaths. Follow-up at 5 years was 100% complete. There were 15 late deaths (96.3 +/- 1.0% survival), including seven of cardiac cause (98.1 +/- 0.7% cardiac survival). Myocardial infarction occurred in 24 patients (94% freedom from myocardial infarction), coronary bypass surgery was required in 63 (84% freedom from bypass surgery), and 365 patients (85%) were asymptomatic at follow-up. At 5 years, 83 patients (20%) had required an additional PTCA. Unstable angina pectoris and proximal left anterior descending coronary artery stenoses were present in 162 patients. The overall survival and cardiac survival in this subset was 94.4 +/- 1.8% and 98.1 +/- 1.1%, respectively. The excellent survival and low event rates over 5 years in this population support the concept that PTCA is safe and effective for patients with symptomatic angina pectoris, single-vessel disease, and normal left ventricular function. PMID- 2964949 TI - Atrial natriuretic factor potentiates forearm reflex vasoconstriction induced by cardiopulmonary receptor deactivation in man. AB - Previous evidence suggests that atrial natriuretic factor (ANF) interferes with the autonomic control of circulation. In the present study we investigated whether ANF modulates forearm vasoconstriction reflexly induced by cardiopulmonary receptor unloading in man. For this purpose, the hemodynamic response to -20 mm Hg lower body negative pressure (LBNP) was assessed under control conditions and during the constant infusion of alpha-human ANF (0.5 micrograms/kg bolus followed by 0.05 micrograms/kg/min) in seven normal subjects. ANF infusion resulted in a slight reduction in blood pressure and right atrial pressure, did not modify heart rate or forearm vascular resistance, but significantly potentiated the reflex increase in forearm vascular resistance during LBNP (+25 +/- 9% under control conditions vs +40 +/- 12% during ANF, p less than .05). In an attempt to clarify the mechanisms underlying the enhanced reflex vasoconstriction during infusion of ANF, in five additional subjects we demonstrated that there was a comparable vascular reflex response to LBNP under control conditions and during nitroglycerin infusion at a dose that induced a reduction in atrial pressure comparable to that observed during ANF. Finally, in seven additional subjects we found that ANF infusion did not alter the reflex hemodynamic responses elicited by carotid baroreceptor unloading induced by a +60 mm Hg increase in external neck pressure. We conclude that during the infusion of a pharmacologic dose of ANF the reflex forearm vasoconstriction in response to selective cardiopulmonary receptor unloading is potentiated. This effect does not seem to be related to the hemodynamic actions of the peptide or to interference with the sympathetic control of peripheral circulation. PMID- 2964950 TI - The effect of coronary angioplasty on coronary flow reserve. AB - To determine the effects of coronary angioplasty on coronary flow reserve (CFR), we studied 32 patients before and immediately after single-vessel coronary angioplasty and 31 patients evaluated late after angioplasty (7.5 +/- 1.2 months, mean +/- SEM). The geometry (percent area stenosis and minimal cross-sectional area) of each lesion was determined by quantitative coronary angiography (Brown/Dodge method) and the integrated optical density was measured by videodensitometry. CFR was measured with a No. 3F coronary Doppler catheter placed immediately proximal to the lesion and a maximally vasodilating dose of intracoronary papaverine. The translesional pressure gradient was obtained in all lesions before and immediately after angioplasty and in 18 of 31 vessels late after angioplasty. CFR immediately after angioplasty returned to normal levels (greater than 3.5 peak/resting velocity ratio) in 14 of 31 patients and was improved, although not normalized, in the remaining 17 patients. CFR immediately after dilation was not significantly correlated with any of the angiographic variables of arterial stenosis nor the resting pressure gradient. Moreover, the pressure gradient and absolute distal coronary pressure at peak hyperemia were not significantly different in vessels with normal and those with abnormal flow reserve immediately after dilation, suggesting that the residual stenosis did not significantly limit hyperemia. Late after angioplasty, however, a significant relationship emerged between CFR and all four indexes of residual arterial stenosis (percent area stenosis r = .70, p less than .01; minimum arterial cross sectional area r = .70, p less than .01; integrated optical density r = .60, p less than .01; and translesional pressure gradient r = .77, p less than .01). Furthermore, in the absence of restenosis, CFR eventually normalized in all patients. These findings demonstrate that in one-half of patients there is a transient reduction in coronary flow reserve immediately after angioplasty. In the absence of restenosis, coronary flow reserve later normalizes. Consequently, measurements of coronary flow reserve immediately after angioplasty may not reflect the eventual success of the procedure in removing physiologic obstruction to coronary blood flow. PMID- 2964951 TI - A rapid and sensitive enzyme immunoassay for Cu/Zn superoxide dismutase with polyclonal and monoclonal antibodies. AB - An enzyme immunoassay for the quantification of human Cu/Zn SOD in serum, urine and erythrocytes was developed applying monoclonal and polyclonal antibodies. The one-step assay is completed within 30 min and enables the detection of 0.3 microgram Cu/Zn SOD per litre. A Cu/Zn SOD concentration of 46 +/- 21.5 micrograms/l and of 1 +/- 0.6 micrograms/mmol creatinine was determined in the serum and the urine, respectively, of healthy individuals. A content of 15 +/- 1.7 ng Cu/Zn SOD was found in 10(6) erythrocytes. Patients with Down's syndrome exhibited a 3.8-fold, a 2-fold and a 1.6-fold higher concentration of Cu/Zn SOD in their serum, urine and erythrocytes. PMID- 2964952 TI - Changes in the plasma hANP level during long-term salt loading in patient with essential hypertension. AB - Serial changes in the plasma hANP (human atrial natriuretic peptide) level in patients with essential hypertension were determined during high salt intake for two weeks after salt restriction for 5 days. The mean plasma hANP level decreased during salt depletion for 5 days from 60 +/- 10 pg/ml to 39 +/- 8 pg/ml. During high salt intake (20 g NaCl/day) for two weeks, the plasma hANP level increased gradually to a maximum of 71 +/- 16 pg/ml on day 7 and then decreased to 54 +/- 9 pg/ml on day 14. The plasma levels of renin activity, aldosterone and noradrenaline decreased during salt repletion. Changes in the plasma hANP level were correlated positively with those of urinary sodium excretion and negatively with those of plasma renin activity on days 7 and 14 of salt repletion. Change in the plasma hANP level correlated with that in the mean blood pressure on day 14, but not day 7 of salt repletion. These findings indicate that the plasma hANP level is closely related to sodium intake in patients with essential hypertension. PMID- 2964953 TI - Reconstructed human epidermis: absence of Langerhans cells and failure to stimulate allogeneic lymphocytes in vitro. AB - Whole human skin can be reconstructed in vitro, using dermal equivalents made of fibroblasts in a collagen matrix. We recently described a new method of epidermalization of dermal equivalents, based on the insertion of punch biopsies and the migration of epidermal cells (EC) on the reconstructed dermis. In the present study, we show that no MHC class II or T6 positive Langerhans cells (LC) can be detected in this new epidermis. Functional studies with EC of this reconstructed epidermis show that these EC completely fail to induce proliferation of allogeneic lymphocytes in mixed epidermal cell lymphocyte reactions and to raise an allogeneic T cell response. In contrast, fresh EC from the same donors induce a strong proliferative and cytotoxic response of the same effector cells. Moreover, the addition of fresh LC-containing EC autologous to effector lymphocytes does not restore an allogeneic proliferative and cytotoxic response directed against class I different EC of the new epidermis. Such a non immunogenic whole skin model composed of two compartments, dermis and epidermis, completely devoid of class II-bearing antigen presenting cells, is thus a very promising technique for allogeneic skin grafting in the treatment of burns. PMID- 2964954 TI - Role of cytokines in the restoration of the delayed-type hypersensitivity reaction of anergic patients. AB - Soluble mediators from peripheral blood lymphocytes activated either by skin test antigens or by alloantigens restored the delayed type hypersensitivity (DTH) reaction in the majority of anergic surgical patients who are at increased risk for sepsis and mortality. Antigen had to be injected together with the mediators and the individual had to be reactive to the antigen for restoration. These results suggest that restoration of the DTH response depends on the ability of cytokines produced and acting in a non-specific manner to promote the response of the anergic patients' specific antigen-sensitive cells to antigen. PMID- 2964955 TI - T cell blast stimulation of MLR: role of interleukin 1 and interleukin 2. AB - T cells blasts were examined for the ability to stimulate primary mixed leucocyte reactions (MLR) in the presence or absence of supplemental interleukin 1 (IL-1) or interleukin 2 (IL-2). After purification, resting T cells were activated using monoclonal anti-CD3 antibody in an accessory-cell free systems. After irradiation, such cells were found to be non-stimulatory in autologous or allogeneic MLR except in the presence of supplemental IL-2. IL-1 was an ineffective cofactor, despite its ability to enhance anti-CD3 mediated stimulation of T cells. It is concluded that T cell blasts may lack an essential factor needed to stimulate IL-2 secretion (other than IL-1), or may elicit responses only from allogeneic T cells not bearing IL-1 receptors. PMID- 2964956 TI - Natural cytotoxicity in the neonate: high levels of lymphokine activated killer (LAK) activity. AB - In 28 healthy full-term newborns the percentage of circulating cells expressing the Leu7 antigen, the marker of natural killer (NK) cells, was significantly lower than in healthy adults. However, newborns and adults did not differ with regard to the percentage of cells reacting with the Leulla, Leullc and TEC NK-1, monoclonal antibodies directed against the IgG Fc receptor of killer cells. Spontaneous NK activity of neonatal cells was profoundly reduced compared to the adult. In contrast, antibody dependent cellular cytotoxicity and NK-like activity generated in mixed lymphocyte cultures were similar in the two groups and lymphokine-activated killer cell (LAK) activity was high in the neonate. Natural killing is thought to play an important role in antiviral immunity since the neonate has a deficient capacity to deal with viral infections. Consequently, the present data indicate either that spontaneous NK is the most informative in vitro measure of newborn natural cytotoxicity in vivo, or, alternatively, that natural killing is not as important in antiviral immunity as previously suggested. PMID- 2964957 TI - Regulation of Fc epsilon receptor expression on a human monoblast cell line U937. AB - Fc epsilon receptor (Fc epsilon R) expression on several human cell lines (U937, RPMI 8866, HL 60, THP-1, and Molt 4) and its regulation were examined by immunofluorescent analysis using a monoclonal anti-human Fc epsilon R antibody, H107. Phorbol ester (PMA), recombinant gamma interferon (IFN-gamma) and H107 itself enhanced Fc epsilon R expression on a FC epsilon R positive cell line U937, whereas these reagents did not induce FC epsilon R expression on the Fc epsilon R negative cell lines, Molt 4, HL 60 and THP-1. Dexamethasone not only suppressed by 50% the spontaneous Fc epsilon R expression on U937 cells but also completely inhibited the enhancement of their Fc epsilon R expression on U937 cells induced by PMA, IFN-gamma or H107. Dexamethasone caused a little suppression of Fc epsilon R expression by RPMI 8866 cells. The results showed that Fc epsilon R expression on a human monoblast cell line U937 was up- or down regulated by a variety of physiological or pharmacological agents. These experimental systems provide a good model for the investigation of the regulatory mechanisms of Fc epsilon R expression. PMID- 2964958 TI - A cellular deficiency in the rheumatoid one-way mixed lymphocyte reaction. AB - Activated lymphocytes expressing transferrin receptors (TFR) are present in the peripheral blood in rheumatoid arthritis. Characterization of these cells shows a CD4 dominance with depressed expression of CD8 and NK antigens, similar to synovial infiltrates. In the present study, one-way mixed lymphocyte reactions were performed to compare the response of rheumatoid patients with that of normal individuals. The TFR+ cells generated were characterized by double-label immunofluorescence. The TFR+ cells from rheumatoid responders showed elevated CD4+ cells and depressed CD8 and Leu-7 (NK) expression compared with normal, throughout the response. This defect was corrected by adding recombinant interleukin 2 at the beginning of the culture period. CD8+ cells stimulated to express TFR following interleukin 2 supplementation of the rheumatoid responses co-expressed HLA-DR. Functional studies indicated that TFR+ CD8+ cells were not cytotoxic, indirectly suggesting them to be suppressor cells. These findings indicate that significant immunoregulatory abnormalities in rheumatoid arthritis may reflect abnormal interleukin 2 biology. PMID- 2964959 TI - Comparative analysis of CD4-4B4 and CD4-2H4 lymphocyte subpopulations in HIV negative homosexual, HIV seropositive and healthy subjects. AB - A comparative analysis of subsets (4B4 and 2H4) within the CD4 lymphocyte subpopulation was made by double marker analysis among 23 healthy heterosexual, 16 healthy HIV seronegative high-risk homosexuals and 82 HIV seropositive subjects. Data show that the significant increase in CD4 lymphocytes observed among seronegative homosexuals corresponds mainly to an increase in CD4-4B4 positive cells while CD4-2H4 subset levels remain comparable to healthy heterosexual controls. A decrease both in CD4-4B4 and CD4-2H4, parallel to a decrease in CD4 subpopulation, was observed in asymptomatic seropositive carriers (SPC) and patients with ARC syndrome (AIDS-related complex) or AIDS (acquired immunodeficiency syndrome). Interestingly, an absolute decrease in CD4 subpopulation in patients with asymptomatic lymph node enlargement (LAS), is chiefly accounted for by a decrease in CD4-4B4 subset. The values of CD4-2H4 subset are significantly higher than those observed for SPC patients and are close to the normal values. These observations are of interest, as no differences between SPC and LAS patients could be detected when CD3, CD4 and CD8 subpopulations were studied. PMID- 2964960 TI - The expression of CD18 is increased on Trisomy 21 (Down syndrome) lymphoblastoid cells. AB - The monoclonal antibody (MAB) 60.3 reacts with the beta chain (CD18) of lymphocyte function associated antigen (LFA-1), encoded by a gene on chromosome 21. We have used 60.3 to measure the expression of CD18 on lymphoblastoid cell lines derived from persons with Trisomy 21 (Down syndrome) by quantitative fluorescence flow cytometric analysis. This showed that CD18 expression is increased on Trisomy 21 compared with normal cells, whereas neither HLA-class I, nor transferrin receptor expression are significantly affected. Similar results were obtained with the CD18 MAB MHM 23, and with the CD11a MAB MHM 24 (LFA-1 alpha). These results suggest that the increase in CD18 expression by Trisomy 21 cells is due to gene dosage, and could influence the immune status of persons with Down syndrome. PMID- 2964961 TI - Circadian rhythms in circulating T lymphocyte subtypes and plasma testosterone, total and free cortisol in five healthy men. AB - Circadian variations of circulating T lymphocyte subtypes and their possible relations with those of endogenous cortisol or testosterone were investigated in five healthy young men. Venous blood (40 ml) was obtained every 4 h for 24 h from each subject in January, March, June, August and November. Leucocyte and differential counts were measured. Mononuclear cells were isolated on Ficoll Paque gradient, and samples were incubated with OKT3, OKT4 or OKT8 monoclonal antibodies for characterizing all T, T helper and T suppressor-cytotoxic lymphocytes respectively. The proportion of labelled lymphocytes was determined under an epifluorescence microscope and the counts of circulating lymphocyte subsets (cells/mm3) computed. Total and free cortisol and testosterone were also determined in the corresponding plasma samples. Results from analysis of variance and cosinor indicated statistically significant differences (P less than 0.001) as a function of both individual subject and circadian sampling time for all variables. Circadian rhythms (with a period, tau = 24 h) were validated for total, T and T helper lymphocytes and for the T helper: T suppressor-cytotoxic ratio (P less than 0.001), with double amplitudes (2A, total extent of variation accounted for by the fitted cosine function) ranging from 25% up to 50% of the 24 h mean (M), and acrophases (phi, time of maximum) localized near 0100 h. A rhythm with tau = 12 h characterized circulating T suppressor-cytotoxic lymphocytes (P less than 0.001; 2A = 36% of M; phi = 0830 and 2030 h). Circadian rhythms were also found for plasma cortisol (either total or free) and testosterone (P less than 0.001). No correlation was found however between time-qualified data of these hormones and the immunological variables herein investigated (162 pairs of data) whether or not a 4 h or an 8 h lag time was considered to allow for hormonal actions to operate. This suggests that neither the circadian organization of the adrenal cortex nor that of the testis play a prominent role in the circadian time structure of the circulation of T lymphocytes. PMID- 2964962 TI - HgC12 induces T and B cells to proliferate and differentiate in BN rats. AB - Mercuric chloride induces in Brown-Norway (BN) rats an autoimmune disease characterized by the production of various autoantibodies and by a marked increase in the IgE serum concentration. This agent is responsible for a T dependent polyclonal activation of B cells, which is probably due to the emergence of autoreactive T cells. The aim of this study was to evaluate the effect of HgCl2 injections on lymphoid organs and on the serum concentration of the various Ig isotypes. HgCl2 induced (1) a lymphoproliferation in spleen and lymph nodes involving B and T helper cells while the number of T suppressor/cytotoxic cells was not modified, (2) an increase in the number of Ig containing cells resulting in a rise in all serum Ig isotypes, and (3) an early thymic atrophy probably immunologically mediated, which was not involved in the induction phase of the disease since adult thymectomy had no effect. These findings demonstrate that the polyclonal effect of HgCl2 is not isotype restricted although the IgE response is predominantly affected and they support evidence for a major role for an excess of T help in the HgCl2-induced polyclonal activation of B cells. It was also observed that B cell areas are present in normal BN rat thymuses, the potential role of which in the induction of autoimmunity remains to be investigated. PMID- 2964963 TI - Mechanism of T cell-derived helper factor production upon stimulation with pokeweed mitogen in humans. AB - T cell-derived helper factors can cause activated B cells to proliferate and differentiate into immunoglobulin (Ig)-secreting cells. In the pokeweed mitogen (PWM)-driven Ig production system, the cell-cell interaction necessary for the production of the helper factors was analysed. T cells were pulsed with PWM with or without autologous monocytes. The T cells were then isolated, and incubated for 48 h. Supernatants were tested for their ability to promote differentiation of activated B cells. We found that cell-cell contact between T cells and monocytes for the first 3 h of culture is required to produce the T cell-derived helper factors upon stimulation with PWM. We next examined which T cell subsets are involved in the production of helper factors and which surface molecules either on T cells or on monocytes are responsible for the cell-cell interaction to produce helper factors. A series of monoclonal antibodies were added to T cell subsets with monocytes during the PWM-pulsed period. Although T4+ and T8+ cells could produce almost equal amounts of helper factor activity upon PWM stimulation, the interactions between T3 and T4 antigens on T4+ cells and Ia-like antigens on monocytes and those between T3 and T8 antigens on T8+ cells and HLA class I antigens on monocytes are respectively essential for the production of T4+ cell-derived and T8+ cell-derived helper factors. PMID- 2964964 TI - Shedding and synthesis de novo of Fc and C3b receptors by cultured guinea-pig macrophages. AB - Resident macrophages freshly obtained from the peritoneal cavity of guinea-pigs were demonstrated to form a higher percentage of Fc and C3b rosettes than elicited macrophages when low concentrations of IgG and IgM-C3b were used to sensitize ox red blood cells (ORBC) in rosette assays. Culture of the total resident and elicited macrophages for 6 h at 37 degrees C resulted in a decrease of Fc and C3b rosette-forming cells, the loss of Fc receptor-bearing cells by resident macrophages only being apparent when using a sub-optimal concentration of sensitizing IgG. After 24 h incubation the percentages of Fc and C3b rosettes returned to their initial values. In contrast, there was no decline in the percentage of Fc and C3b rosettes formed by the adherent population of resident and elicited macrophages cultured for 6 h. However, extending the incubation of the adherent macrophage to 24 h produced an increase of Fc receptor-positive cells and a dramatic decrease of C3b receptor-positive cells. Culture supernatants of the total macrophage population that had been incubated for 6 h inhibited Fc and C3b rosette formation by freshly obtained elicited macrophages. These results, together with the demonstration that treatment of the total macrophage population with cycloheximide led to an inhibition of Fc and C3b receptor expression after 24 h culture, suggest that the Fc and C3b receptors of guinea-pig macrophages are shed and synthesized de novo during short-term culture. This system could be applied to the study in vitro of soluble immunoregulatory mediators on macrophage functions which are dependent on the expression of Fc and C3b receptors. PMID- 2964965 TI - Effect of opiate, general anaesthesia and surgery on plasma atrial natriuretic peptide levels in man. AB - 1. Animal data suggest that opiates, halothane anaesthesia and activation of the sympathetic system stimulates release of atrial natriuretic peptide (ANP). To examine whether this is so in man, venous ANP levels were measured in five patients undergoing elective cholecystectomy. 2. Plasma levels of cortisol, aldosterone, norepinephrine and epinephrine increased 3-6 fold during the study. Cortisol-aldosterone relationships were close in all patients (r = 0.73-0.97), whereas plasma renin activity and aldosterone correlations were strong in only two subjects. 3. Baseline plasma ANP concentrations were within the normal range and were not altered by opiate injection, anaesthesia, or surgery. 4. Unlike experimental animals, man exhibits little or no ANP response to opiates, halothane, or surgical stimulation of the sympathetic nervous system. PMID- 2964966 TI - Atrial natriuretic peptide release in the rabbit is independent of cardiac nerve activity. AB - 1. Changes in plasma atrial natriuretic peptide (ANP) were examined in conscious rabbits in response to a 33% blood volume expansion in intact animals and after blockade of cardiac nerve activity. 2. Blood volume expansion by one-third markedly increased right atrial pressure and resulted in a four-fold increase in plasma ANP. 3. Cardiac nerve blockade with intrapericardial procaine had no effect on resting plasma ANP levels. The ANP responses to volume expansion in the presence of cardiac nerve blockade were similar to those seen in intact animals. 4. Release of ANP from its cardiac stores in response to volume expansion is not influenced by cardiac nerve activity. PMID- 2964967 TI - Immunohistochemical analysis of intrathyroidal lymphocytes in Graves' disease: evidence of activated T cells and production of interferon-gamma. AB - Previous immunohistochemical studies of the thyroid gland in Graves' disease (GD) have demonstrated that (i) lymphoid cells infiltrating the interstitium of the gland predominantly express the CD4+ (T-helper) phenotype; (ii) lymphoid cells infiltrating the epithelial compartment express predominantly the CD8+ antigen; and (iii) thyroid epithelial cells (thyrocytes) express class II histocompatibility antigens, possibly in response to endogenous production of interferon-gamma (IFN-gamma) by the infiltrating lymphocytes. In the present study the infiltrating lymphoid cells and thyrocytes in GD were further characterized by immunocytochemistry. Scattered infiltrating interstitial lymphocytes stained positively with antibodies directed against IFN-gamma and the receptor for interleukin 2. These lymphocytes were most prominent around the periphery of lymphoid nodules. The interstitial lymphocytes predominantly expressed the CD4+ 2H4+ phenotype rather than the complementary CD4+ 4B4+ phenotype, suggesting the presence of the suppressor-inducer rather than helper inducer T cell subset. The infiltrating lymphocytes were associated with thyrocytes expressing HLA-DR antigens, as previously reported, and also HLA-DQ antigens, but to a lesser extent. The presence of HLA-DQ may also suggest the possibility of an induced suppressor response. The intraepithelial CD8+ cells did not stain with antibodies to CD3, 2H4, 4B4, or Leu 7 and their identity remains unknown. PMID- 2964969 TI - Adult scoliosis. Current concepts of treatment. AB - The treatment of adult patients with idiopathic scoliosis is a challenge of the 1980s. Surgical treatment is difficult, and the complication rate is high. Nevertheless, surgical treatment may offer the best alternative when the specific problem has been defined, the goal is realistic, the technical solution is apparent, the alternatives have been considered, and the benefits outweigh the risks. PMID- 2964968 TI - Studies of lymphocyte subpopulations and immunoglobulin production in IgA nephropathy. AB - This work was undertaken to examine the immunoregulation of T lymphocytes in patients with IgA nephropathy. Fifty patients and thirty-seven control subjects were studied in an infection-free interval. T lymphocyte subpopulations were determined using OKT monoclonal antibodies against helper (OKT4) and suppressor (OKT8) T cell subsets. The proportions of T lymphocyte subpopulations did not differ between patients and controls. Patients with significant renal impairment demonstrated a reduced OKT4/T8 ratio (p less than 0.001) due to an absolute reduction of T helper cells (p less than 0.02) and an increase of T suppressor cells (p less than 0.001). Longitudinal studies performed in 13 patients revealed consistent findings during clinical quiescence. However, synpharyngitic macroscopic haematuria was associated with a rise in T4 positive cells and a simultaneous reduction of T8 positive cells; these changes reverted to pre infection values when the infection subsided. Functional studies assessing T lymphocyte activities including in vitro immunoglobulin synthesis by cultured peripheral blood mononuclear cells, thymidine uptake by cultured lymphocytes and T lymphocytes activation with expression of interleukin-2 receptors were measured in unstimulated and mitogen stimulated cultures. No significant difference between patients with IgA nephropathy in clinical quiescence and the control subjects was demonstrated. Our results failed to support a shift in the immuno regulatory T lymphocyte subpopulations during clinical quiescence but a more profound defect in immunoregulation may probably occur during clinical exacerbation. PMID- 2964970 TI - Brucellosis: appearance on skeletal imaging. AB - Brucellosis is an endemic disease in the Middle East. Its incidence in Kuwait has increased during the last 5 years. Bone and joint involvement causes major symptoms and disabilities. Radionuclide bone scans are more sensitive than radiographs in detecting these lesions. The aim of this study is to describe the abnormal patterns detected on bone imaging in acute and chronic brucellosis. Tc 99m MDP bone scans of 56 patients with established diagnosis of brucellosis (19 acute and 37 chronic) were retrospectively analyzed. Bone scans were positive in 8 of 19 patients (42%) with acute brucellosis and in 28 of 37 patients (76%) with chronic brucellosis. Six patterns were observed: involvement of an entire body of one or more vertebrae, especially at the lumbar region (50%); sacroiliitis (41%); focal high uptake at the junction of the upper and lateral margins of the vertebra "Caries sign" (27%); multiple costovertebral joints and costochondral junction involvement (19%); involvement of large joints similar to degenerative osteoarthritis (25%); and focal involvement of long bone (11%). PMID- 2964971 TI - Calf muscle adaptation in intermittent claudication. Side-differences in muscle metabolic characteristics in patients with unilateral arterial disease. AB - The adaptation of enzyme activities, notably in the oxidative metabolism, and of prerequisites for tissue transport of oxygen in the claudication leg was evaluated by comparing muscle biopsies from the gastrocnemius muscle of the claudication and the symptom-free leg of seven patients with unilateral claudication. The claudication leg had higher activities of a marker enzyme for mitochondrial oxidative capacity, citrate synthase (CS), as well as of the MB and the mitochondrial isoenzyme of creatine kinase (CK), which are considered to be involved in the transfer of high energy phosphate from the mitochondria to the resynthesis of ATP in the cytoplasm. The difference between claudication and healthy leg in activities of these CK isoenzymes were well correlated with the corresponding side difference in CS activity. No significant differences between claudication and healthy leg were found in distribution of muscle fibre types or fibre dimension, capillary density or myoglobin content, nor was there any side difference in phosphofructokinase or lactate dehydrogenase. Side differences tended to be greater in those patients with the most advanced obstructive arterial disease as estimated from non-invasive pressure measurements. It is concluded that in reasonably physically-active patients, the mode of ischaemia to which the claudication leg is subjected leads to a metabolic adaptation characterized by increased activities of enzymes involved in the oxidative metabolism, but no significant adaptation of either the conditions for local oxygen transport, as estimated by myoglobin content, and capillary density, or capacity for anaerobic metabolism. PMID- 2964972 TI - Changes in atrial natriuretic factor during preload reduction with nitroglycerin in patients with congestive heart failure. AB - Nine patients with congestive heart failure, New York Heart Association class II III, were evaluated with right heart catheterization. Plasma atrial natriuretic factor (ANF) was determined in blood samples from the pulmonary artery simultaneously with recordings of right atrial, pulmonary arterial, pulmonary capillary wedge and systemic arterial pressures and heart rate during preload reduction with 0.5 mg nitroglycerin sublingually. Basal plasma ANF levels were higher in patients with congestive heart failure compared to normal controls, and correlated to right atrial, pulmonary arterial, and pulmonary capillary wedge pressures. After nitroglycerin all patients had reductions in right atrial, pulmonary arterial, and pulmonary capillary wedge pressures and a simultaneous decrease in plasma ANF concentrations, reaching lowest values after 10 min. Central pressures and plasma ANF rose to baseline values within 30 min. After nitroglycerin plasma ANF concentrations correlated to pulmonary arterial and pulmonary capillary wedge pressures, while changes in plasma ANF correlated to changes in right atrial and pulmonary arterial pressures. These results provide further evidence that ANF is released by a pressure-sensitive mechanism and demonstrates that ANF secretion in relation to central pressure variations is preserved in patients with congestive heart failure and that the response is rapid. PMID- 2964973 TI - Clinical trials--time for a paradigm shift? PMID- 2964974 TI - New approaches to the pathogenesis and treatment of diabetic microangiopathy. PMID- 2964975 TI - Is serum fructosamine a clinically useful test? PMID- 2964976 TI - Androgen levels in men with diabetes mellitus. AB - Patients in varying degrees of diabetic ketoacidosis had low serum testosterone levels which rose following recovery (5.4 +/- 1.3----13.7 +/- 1.7 nmol/l, p less than 0.02, mean +/- SE). Blood glucose control of 30 non-ketotic diabetic men (15 insulin-dependent and 15 non-insulin-dependent) did not correlate significantly with serum testosterone concentrations. Although previous authors had suggested that the adrenal androgen status of diabetic patients might be abnormal, adrenal androgens androstenedione and dehydroepiandrosterone sulphate (DHEAS) were similar in our 30 men to age-matched healthy men. PMID- 2964977 TI - Microalbuminuria: a major risk factor in non-insulin-dependent diabetes. A 10 year follow-up study of 503 patients. AB - The impact of microalbuminuria on mortality as well as other risk factors was investigated in a 10-year follow-up study of 503 predominantly non-insulin dependent diabetic patients of whom 265 had died. Using Cox's regression analysis the prognostic influence of age, sex, age at diagnosis, known diabetes duration, blood pressure, fasting plasma glucose, relative weight, serum creatinine, retinopathy, and treatment was evaluated as well as morning urine albumin concentration (UAC) in four categories, i.e. UAC less than or equal to 15 micrograms/ml (normal), 15 micrograms/ml less than UAC less than or equal to 40 micrograms/ml, 40 micrograms/ml less than UAC less than or equal to 200 micrograms/ml and UAC greater than 200 micrograms/ml. Age, UAC, known duration, and serum creatinine were the only significant risk factors. After correction for the other three independent risk factors, the hazard ratios in the elevated UAC categories relative to the group with UAC less than or equal to 15 micrograms/ml were 1.53 (p = 0.007), 2.28 (p = 0.000002), and 1.82 (p = 0.02). The statistically significant correlations with UAC were: age (r = 0.09, p less than 0.05), duration (r = 0.14, p less than 0.01), systolic blood pressure (r = 0.12, p less than 0.01), serum creatinine (r = 0.33, p less than 0.001), and fasting plasma glucose (r = 0.12, p less than 0.01). Increased UAC was associated also with retinopathy (p = 0.01). Fifty-eight per cent of the deaths were caused by cardiovascular disease or stroke; only 3% died from uraemia. A reinvestigation including blood pressure, fasting plasma glucose, and UAC was made on 208 survivors. PMID- 2964978 TI - The Medi-Jector II: efficacy and acceptability in insulin-dependent diabetic patients with and without needle phobia. AB - Efficacy and acceptability of the Medi-Jector II, compared with conventional syringes, was evaluated in a cross-over study (two 4-week periods) in 14 adult insulin-dependent diabetic patients. In 6 out of these 14 patients the clinical diagnosis of needle phobia was confirmed by anxiety tests for trait (45.8 +/- 12.8 (SD) versus 34.0 +/- 6.2; p less than 0.05) and state (50.3 +/- 10.9 versus 28.9 +/- 5.2, p less than 0.05). Five patients (one needle-phobic) dropped out, all for Medi-Jector-related problems. No significant differences were found in body weight, insulin dosage, and number of hypoglycaemic reactions. HbA1 after the Medi-Jector period was significantly higher than after the conventional period (9.8 +/- 1.2 (SE) versus 9.1 +/- 1.1 (SE)%; p less than 0.05). The questionnaire, evaluating the Medi-Jector, revealed poor acceptance of the device (7/9 patients' general impression being moderate or bad). The use of the Medi Jector in comparison with conventional injections caused the patients to complain about: more immediate pain (4/9 patients), delayed pain (often: 4/9, sometimes: 2/9), more insulin leakage (6/9), more bleeding (6/9), and more haematomas (5/9). Except for 'often occurring delayed pain' (4/5 needle-phobics versus 0/4 non needle-phobics; p less than 0.05), needle-phobic patients scored the questionnaire similarly to the other patients. We conclude that the use of the Medi-Jector II did not cause a major short-term loss of metabolic control, but that the device was not well accepted by our patients, independent of the existence of needle phobia. PMID- 2964979 TI - Detection of antibodies against wheat germ agglutinin bound glycoproteins on the islet-cell membrane. AB - An attempt was made to detect islet cell surface antibodies (ICSAb) using solubilized islet-cell glycoproteins as antigens. Isolated rat islets were labelled with 125I-wheat germ agglutinin (WGA) and solubilized by Nonidet P-40 with sonication. 125I-WGA-bound islet-cell proteins were incubated with test sera, and bound antibodies were precipitated with anti-human IgG or IgM immunobeads. Serum which had a bound percent beyond the mean plus 2SD of control sera was defined as antibody-positive. Results obtained by this method correlated well with those by the immunofluorescence method of detecting ICSAb. Prevalences of antibodies were 21/114 (18%) for IgG antibodies and 9/114 (8%) for IgM antibodies in patients with insulin-dependent diabetes mellitus (IDDM). The prevalence was highest for both IgG and IgM antibodies in patients within a year of the onset of disease (38 and 25%, respectively), and decreased thereafter. The prevalence of IgM antibodies was lower than that of IgG antibodies at all stages. In NIDDM patients, the prevalence of antibodies was 5/72 (7%) for both IgG and IgM antibodies. If these preliminary results are confirmed, this radioassay may be developed to detect antibodies against islet cell membrane proteins on a large scale. PMID- 2964980 TI - Elevated urinary prostaglandin excretion and the effect of indomethacin on renal function in incipient diabetic nephropathy. AB - We investigated whether the glomerular synthesis of prostaglandins modulates the glomerular filtration rate and urinary albumin excretion in incipient diabetic nephropathy (defined as urinary albumin excretion between 30 and 300 mg/24 h (microalbuminuria) in two out of three sterile ketone-free 24-h urine collections in patients having insulin-dependent diabetes mellitus (IDDM) without hypertension or other kidney disease). The urinary excretion of prostaglandin E2 was significantly elevated in 8 insulin-dependent diabetic patients with incipient nephropathy as compared with 9 normoalbuminuric IDDM patients and 11 healthy controls: 317 (182-1273); 95 (67-225); 132 (54-263) pg/min, respectively (2p less than 0.01). Glomerular filtration rate (single bolus 51Cr-EDTA technique) and albuminuria (radioimmunoassay) were measured twice within 2 weeks in 8 females having IDDM with incipient nephropathy. The study design was a randomized double-blind trial with the patients receiving either indomethacin (150 mg/day) or placebo for 3 days prior to the kidney function studies. Indomethacin treatment induced a significant reduction in urinary prostaglandin E2 excretion (73%) (2p less than 0.01), urinary albumin excretion rate diminished from 207 (63-253) to 87 (49-147) mg/24 h (2p less than 0.01), fractional clearance of albumin declined (70%) (2p less than 0.01). Glomerular filtration rate remained stable (108 (88-133) versus 110 (95-142) ml/min). Blood glucose and blood pressure were comparable during the placebo and indomethacin treatment (12.6 +/- 3 versus 13.4 +/- 5 mmol/l and 122/79 +/- 3/9 versus 122/82 +/- 4/10 mmHg, respectively). Our results suggest that enhanced glomerular synthesis of vasodilating prostaglandins may accelerate microalbuminuria in incipient diabetic nephropathy. PMID- 2964981 TI - Comparison of continuous subcutaneous insulin infusion with multiple insulin injections using the NovoPen. AB - Twenty-one patients with insulin-dependent diabetes mellitus (IDDM) participated in a 20-week randomized cross-over comparison of continuous subcutaneous insulin infusion (CSII) with intensified conventional treatment (ICT) using the NovoPen. The Medix or the Auto-Syringe pumps were used for CSII and, during ICT with NovoPen, conventional plastic syringes were used for injections of intermediate acting insulin at bedtime. At entry HbA1c, was 8.7 +/- 0.4% (mean +/- SE) in CSII patients and 8.8 +/- 0.5% in the ICF group. HbA1c declined significantly in both groups (ICT 7.6 +/- 0.2%; CSII 7.6 +/- 0.2%) though there was no significant difference between the responses. Overall mean blood glucose was slightly but significantly lower during CSII than during ICT (CSII: 7.6 +/- 0.2 mmol/l; ICT: 8.7 +/- 0.4 mmol/l, p less than 0.05). The number of hypoglycaemic episodes did not differ significantly between patients treated with NovoPen and CSII. At the end of the study, a questionnaire revealed that all but one patient preferred ICT with NovoPen to conventional therapy. Given the choice for future treatment, 6 patients chose CSII, 12 patients preferred ICT with NovoPen and 1 was unsure. PMID- 2964982 TI - UK Prospective Diabetes Study. IV. Characteristics of newly presenting type 2 diabetic patients: male preponderance and obesity at different ages. Multi-center Study. AB - The characteristics of newly presenting Type 2 diabetes mellitus have been examined in 1857 newly diagnosed diabetic patients aged 25-65 years inclusive. The males were less obese than the females (121% vs 141% IBW, respectively), but a male-dominated sex ratio of 1.54 was found. Taking into account the prevalence of obesity in the general population, males had a 2.5-fold relative risk of presenting with diabetes, although with increasing obesity the male preponderance was lost. Presentation increased with age up to the age of 55 years. Patients presenting at all ages had similar glycaemia and were similarly obese. Those presenting at a younger age were usually particularly obese in relation to the general population. Obese patients were less physically active than normal weight patients. Type 2 diabetes had a seasonal variation of presentation with a peak in January to April. PMID- 2964983 TI - Geographical variation in the incidence of diabetes mellitus in Scottish children during the period 1977-1983. AB - Routinely collected Scottish hospital discharge data were used to identify newly diagnosed cases of diabetes mellitus under the age of 19 years. In the period 1977-83 there were 2183 cases, an average annual incidence of 21.0 per 100,000 per annum. This rate is one of the highest reported and represents a continuation of the increase in incidence which occurred during the period 1968-76. Statistically significant geographical variation in incidence was observed with the higher incidence in the least densely populated areas. Seasonal variation in the month of first admission was evident in all except the under-five age group with peaks occurring in winter. An estimated 60% of newly diagnosed cases were readmitted within 5 years and the pattern of readmission was found to differ little between males and females or between age groups. PMID- 2964986 TI - Statement on post-prandial or reactive hypoglycaemia. PMID- 2964984 TI - Serum lipids, lipoproteins and macrovascular disease in non-insulin-dependent diabetics: a possible new approach to prevention. AB - The relationship between macrovascular disease and serum lipids, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL), and subfraction cholesterol, and apolipoproteins has been examined in 53 female and 95 male patients with non-insulin-dependent diabetes mellitus (NIDDM). In males, those with macrovascular disease had higher serum and LDL cholesterol concentrations than those without. In females, those with macrovascular disease had higher levels of serum triglyceride, cholesterol, LDL cholesterol, as well as lower HDL, HDL2, and HDL3 cholesterol and apoprotein A-1, than those without. On multivariate analysis, LDL cholesterol was the most important association with macrovascular disease in males and apoprotein A-1 in females. In a subgroup of 36 patients, a double-blind placebo controlled study using bezafibrate or placebo, in addition to conventional oral hypoglycaemic therapy over 4 months, showed falls in serum and LDL cholesterol and in serum triglyceride and a rise in HDL cholesterol in the treated group. These changes should reduce the incidence of macrovascular disease in NIDDM and we suggest further prospective studies of such therapy in addition to conventional oral hypoglycaemic agents. PMID- 2964987 TI - Contact dermatitis from N-(alpha-chlorobenzylidene)phenylhydrazine. AB - N-(alpha-chlorobenzylidene)phenylhydrazine (AC) is both a strong irritant and a powerful sensitizer. 5 out of 6 people at risk fell ill after only once coming into contact with this halogenated hydrazine derivative. As in the first report concerning this problem the compound was used to train chemistry students. PMID- 2964985 TI - Non-insulin-dependent diabetes and renal replacement therapy. AB - Reports of renal replacement therapy in diabetes usually refer to patients with insulin-dependent diabetes mellitus (IDDM) only, and little is known about renal failure in non-insulin-dependent diabetics (NIDDM). A high proportion, 46/141 (32%), of the diabetics treated at our unit since 1974 had NIDDM. They were older at treatment (56 +/- 9 years, mean +/- SD) compared to the IDDM patients (39 +/- 10 years, p less than 0.001), and had a shorter duration of diabetes (13 +/- 8 years versus 23 +/- 8 years, p less than 0.001). Asians and Afro-Caribbeans accounted for 48% of the NIDDM patients (22/46) compared to only 7% of those having IDDM (6/95, p less than 0.0001). Non-diabetic renal disease accounted for the renal failure in 32% (15/46) of the NIDDM patients but only in 10.5% (10/95) of the IDDMs (p less than 0.001). Despite these differences the prevalence of other diabetic complications (retinopathy, neuropathy, and cardiovascular disease) was similar. Patient survival after transplantation was poorer in NIDDM than IDDM (23% and 57%, respectively, at 2 years). Survival on dialysis was equally poor in NIDDM and IDDM. Thus, NIDDM patients treated for renal failure are more commonly non-European and more often have non-diabetic renal disease. Yet other diabetic complications occur to the same extent in both IDDM and NIDDM patients with diabetic nephropathy. PMID- 2964988 TI - Bioengineering and the patch test. AB - Several non-invasive techniques based on different physical principles have been developed to investigate skin function and have been used for patch test assessment. In the present paper, the advantages and the defects of these methods are described in the light of the more recent data available in literature. Meanwhile, the eye and the fingers still remain the simplest method to assess skin irritancy. Bioengineering techniques could provide efficient recording systems for monitoring skin color, skin blood flow and barrier function damage, useful to the investigator rather than to the clinician. PMID- 2964989 TI - Allergic and irritant skin reactions evaluated by laser Doppler flowmetry. AB - Skin blood flow (SBF) in allergic and irritant reactions was determined by laser Doppler flowmetry (LDF) in an attempt to differentiate these reactions in an objective way. 12 subjects with known allergy to nickel were patch tested with nickel sulphate, nickel chloride, and with sodium lauryl sulphate (SLS) 1, 2, 5 and 10%. A positive correlation between the concentration of SLS and SBF was found. Compared with control sites, SBF in weak (+) and strong (++) positive allergic and irritant reactions was increased approximately 5-10 fold 1 and 2 days after application, and decreased towards normal 7 days after application. At day 7, the mean SBF was higher in allergic reactions than in irritant reactions. However, no consistent differences were found. It is concluded that although LDF can be used to quantify the strength of allergic and irritant skin reactions, this method cannot be used solely to separate these 2 types of reaction. PMID- 2964990 TI - Occupational dermatitis from colophony. PMID- 2964991 TI - Contact dermatitis due to amcinonide. PMID- 2964992 TI - A multicentered pharmacokinetic, pharmacodynamic study of once-a-month injectable contraceptives. I. Different doses of HRP112 and of DepoProvera. World Health Organization Task Force on Long-acting Systemic Agents for Fertility Regulation. AB - A multicentered study was undertaken at three institutions in Hungary, Mexico and Thailand in women of reproductive age to evaluate the effects of full and half doses of HRP112 (depot-medroxyprogesterone acetate (DMPA) plus estradiol cypionate) and DMPA alone on ovarian function, bleeding patterns and HDL cholesterol levels. Full dose HRP112 contained 25mg DMPA plus 5mg, of estradiol cypionate and the half dose, 12.5mg DMPA plus 2.5mg of estradiol cypionate. The full and half dose DMPA were 25 and 12.5mg respectively. In all, 88 women were recruited in the study and randomized within each centre, to the four treatment groups. Subjects were studied for a control cycle, three one-month injection intervals and followed-up for a further two months. Serum concentrations of estradiol, progesterone and medroxyprogesterone acetate were determined three times a week during the third injection interval and during the two months of follow-up. While the results from all centres indicated that the four preparations were all effective in inhibiting ovulation for at least one month, there were marked between centre differences in pharmacokinetic profiles. More regular bleeding patterns were observed in women who received the estrogen progestogen combination preparations than in those who received DMPA alone. PMID- 2964993 TI - Thrombogenicity markers in clinical and ex vivo assessment of membrane biocompatibility. PMID- 2964994 TI - Idiosyncratic drug-induced thrombocytopenia. PMID- 2964995 TI - The effect of nebulized bronchodilator therapy on intraocular pressures in patients with glaucoma. AB - A controlled double-blind crossover study of ocular complications associated with nebulized ipratropium bromide and salbutamol therapy for respiratory distress was undertaken in 46 chronic bronchitis patients. There was no significant rise in intraocular pressure or change in anterior chamber angle in patients with open angle glaucoma, narrow-angle glaucoma or control subjects following treatment with either drug. However, when the two drugs were used in combination, intraocular pressure rose in patients with narrow-angle glaucoma but not in patients with open-angle glaucoma or in control subjects. Transient angle closure was seen in five of these patients. Intraocular pressures did not rise when swimming goggles were used to protect the eyes or when antiglaucoma treatment was continued. Nebulized bronchodilator therapy is safe in nonglaucomatous patients and those with open-angle glaucoma. Ocular complications can follow combined ipratropium bromide and salbutamol nebulization in patients with narrow-angle glaucoma, but can be prevented by using the drugs separately, protecting the eyes and ensuring continued antiglaucoma measures. PMID- 2964996 TI - Electrocardiographic criteria for the diagnosis of right ventricular hypertrophy verified at autopsy. AB - Four commonly used sets of electrocardiographic (ECG) criteria for diagnosing right ventricular hypertrophy (RVH) were tested for sensitivity and specificity in 12 men with isolated RVH, 15 with combined right and left ventricular hypertrophy and 24 with normal ventricular weights. The cardiac ventricles were weighed separately at autopsy and the left-to-right ratio was calculated. All the patients showing isolated RVH, with a left-to-right ratio of two or less, had died of respiratory causes. Three sets of ECG criteria showed a specificity of 100 percent, but sensitivities only from 26 to 44 percent. One set was more sensitive (74 percent) but much less specific than the others (79 percent). A new combination of ECG criteria attained a 63 percent sensitivity and a 96 percent specificity. PMID- 2964997 TI - Renal, cardiovascular and hormonal effects of alpha-human atrial natriuretic peptide on essential hypertension. PMID- 2964998 TI - Experimental study on cytotoxic effects of hyperbaric oxygen and photodynamic therapy on mouse transplanted tumor. PMID- 2964999 TI - Genetics of coronary heart disease and its risk factors. AB - Lipoprotein parameters related to coronary heart disease (CHD) exhibit impressive heritability, and several traditional genetic marker systems are associated with lipid levels or CHD. Recent studies indicate a population-attributable risk of 28% for myocardial infarction for men below age 60 in the top quartile of Lp(a) lipoprotein levels. Thus, a high level of Lp(a) lipoprotein emerges as a major genetic risk factor for premature CHD. Studies of DNA polymorphisms at apolipoprotein loci have uncovered associations with lipid levels and genetic linkage between DNA polymorphisms at the apolipoprotein B (apoB) locus and the Ag(x) antigenic polymorphism of low density lipoprotein. This finding proves that the Ag(x) antigenic variation resides in apoB and co-assigns its locus to chromosome 2. Lipid associations of the Ag(x) polymorphism and of DNA polymorphisms at the apoB locus are internally consistent and consistent with association between Ag(x) and DNA variants. A new approach to the study of gene environment interactions, using monozygotic twin pairs makes it possible to uncover genes that contribute to the frame within which lifestyle factors can cause changes in a clinically relevant quantitative parameter such as serum cholesterol concentration. A new concept of interaction between 'level genes' and 'variability genes' in the aetiology of atherosclerosis emerges from these studies. PMID- 2965000 TI - Apolipoproteins, quantitative lipoprotein traits and multifactorial hyperlipidaemia. AB - Genetic polymorphism and rare mutants of apolipoproteins occur in humans. The polymorphism of apolipoprotein E (apoE) is controlled by three common alleles, epsilon 2, epsilon 3, and epsilon 4, which code for proteins that differ in lipoprotein receptor binding activity, or in their catabolism in vivo, or both. This may explain the observed significant effects of the apoE alleles on the phenotypic variance of plasma lipoprotein concentrations in different ethnic groups and, moreover, the involvement of apoE alleles in the pathogenesis of multifactorial forms of hyperlipidaemia, for example, hypertriglyceridaemia, familial type III hyperlipidaemia (apoE-2 Arg-158----Cys) and polygenic hypercholesterolaemia (apoE-4 Cys-112----Arg). A further polymorphic gene locus controls the concentrations of the Lp(a) lipoprotein complex in plasma, which may vary from less than 1 mg/dl to greater than 200 mg/dl between different individuals. This lipoprotein contains two different polypeptides, apoB-100 and the Lp(a) glycoprotein. The Lp(a) glycoprotein exhibits genetic polymorphism which is controlled by a series of autosomal alleles at a single locus and which is associated with lipoprotein concentrations in plasma. This suggests that the same gene locus is involved in determining Lp(a) glycoprotein phenotypes and Lp(a) lipoprotein concentrations in plasma. Thus, there is evidence that variability in apolipoprotein genes relates to the normal variance of lipoprotein concentrations in the population and that this variability is a major genetic factor in multifactorial forms of hyperlipidaemia. PMID- 2965002 TI - Severe hypersensitivity reaction upon rechallenge with trimethoprim sulfamethoxazole in a patient with AIDS. AB - A patient with acquired immunodeficiency syndrome (AIDS) developed rash, fever, neutropenia, and elevated liver function tests during an initial course of trimethoprim-sulfamethoxazole (TMP-SMX) therapy. Upon reexposure to the drug, the patient experienced a severe anaphylactoid reaction associated with pulmonary edema and rhabdomyolysis. Reactions associated with TMP-SMX rechallenge in this patient population have been previously reported but have not been associated with this degree of severity. TMP-SMX therapy should be instituted with extreme caution in patients with AIDS who have demonstrated a prior hypersensitivity reaction to the drug. PMID- 2965001 TI - Beta-endorphin-like immunoreactivity in normal mucosa, muscle layer, adenocarcinoma, and polyp of the colon. AB - beta-Endorphin-like immunoreactivity was detected in the mucosa and muscle layer of normal colon, adenocarcinomas derived from the colon mucosa, and colon polyps which were histologically confirmed to be adenoma without a focus of carcinoma or with in situ carcinoma. The contents of beta-endorphin-like immunoreactivity in adenocarcinomatous tissue (11.94 +/- 1.77 pmol/g wet wt) and colon polyps without focus of carcinoma (10.71 +/- 1.50 pmol/g wet wt) were found to be significantly higher than those in the mucosal layer (6.86 +/- 0.64 pmol/g wet wt) and muscle layer (8.30 +/- 0.68 pmol/g wet wt) of normal colon. These data suggest that the production of beta-endorphin-like immunoreactivity is specifically increased in some adenocarcinomas and adenomatous polyps and may be related to the alteration of bowel habits. Gel exclusion chromatography of beta-endorphin-like immunoreactivity revealed three peaks corresponding to beta-endorphin, beta lipotropin, and an immunoreactive form between the two. In the mucosal layer and muscle layer of the colon, a broad major peak was eluted at the position of beta endorphin, and minor peaks were eluted at the position of beta-lipotropin and between beta-endorphin and beta-lipotropin. In adenocarcinoma and polyp, the peak size corresponding to authentic beta-lipotropin was greater than that of beta endorphin. This study demonstrated that beta-endorphin-like immunoreactivity existed at a high concentration in some colon adenocarcinomas and polyps whose elution patterns were different from those of normal colon tissue. PMID- 2965003 TI - Tocainide: a severe adverse reaction. AB - Tocainide is a primary analog of lidocaine with antiarrhythmic properties used to treat ventricular rhythm disorders. A 76-year-old man with benign paroxysmal premature ventricular contractions was treated with tocainide and developed a generalized maculopapular lupoid eruption, bleeding from the lips and gingivae, vertigo, gross tremors of the extremities, fever, and short-term memory loss, which required hospitalization. The patient recovered slowly over three months with no permanent sequelae after discontinuing the drug and receiving rigorous supportive care. His excellent physical status and absence of concomitant illness contributed to an uneventful recovery. Tocainide is a potent cardioactive drug with a long biological half-life and should be used with caution. PMID- 2965004 TI - [Current complication rate of percutaneous transluminal coronary angioplasty in stable and unstable angina]. AB - During a four-year period (1983-1986) percutaneous transluminal coronary angioplasty (PTCA) was performed on 930 patients with stable or unstable angina with a mortality rate of 0.4%. A transmural myocardial infarct developed in 1.1% and 1.0% of patients required an urgent aorto-coronary bypass. Thus the total rate of severe cardiac complications was 2.5%. Compared with the years 1983-1985, there was in 1986 a significant fall in the number of deaths and of myocardial infarcts from 2.2% to 0.5% (P less than 0.05), while there was a nonsignificant increase in emergency coronary bypass surgery from 0.7 to 1.3%. Patients with unstable angina compared with those with stable angina had a significantly higher mortality rate (1.0% vs. 0.2%; P less than 0.05), incidence of infarction (2.0% vs. 0.6%; P less than 0.05), and emergency operations (2.0% vs. 0.5%; P less than 0.05). The total risk of a severe cardiac complication was 1.2% for stable and 5.2% for unstable angina (P less than 0.001). PMID- 2965005 TI - Myocardial salvage: angioplasty and coronary artery bypass. AB - Percutaneous transluminal coronary angioplasty has shown great promise in the treatment of acute myocardial infarction, both alone and in combination with thrombolysis. Because of time constraints, intravenous thrombolysis probably will emerge as the initial therapy of choice in most settings. Because thrombolysis often has a high incidence of reocclusion and recurrent ischemia, angioplasty will continue to play a role in relieving residual coronary artery stenosis. Coronary artery bypass surgery is not often used currently as sole therapy for acute myocardial infarction. Nonetheless, like angioplasty, bypass surgery plays a supporting role in relieving post-thrombolytic stenosis, as well as in treating complications of angioplasty. PMID- 2965006 TI - The sequence of rat leukosialin (W3/13 antigen) reveals a molecule with O-linked glycosylation of one third of its extracellular amino acids. AB - Leukosialin is one of the major glycoproteins of thymocytes and T lymphocytes and is notable for a very high content of O-linked carbohydrate structures. The full protein sequence for rat leukosialin as translated from cDNA clones is now reported. The molecule contains 371 amino acids with 224 residues outside the cell, one transmembrane sequence and 124 cytoplasmic residues. Data from the peptide sequence and carbohydrate composition suggest that one in three of the extracellular amino acids may be O-glycosylated with no N-linked glycosylation sites. The cDNA sequence contained a CpG rich region in the 3' coding sequence and a large 3' non-coding region which included tandem repeats of the sequence GGAT. PMID- 2965007 TI - Identification of an E1A-inducible cellular factor that interacts with regulatory sequences within the adenovirus E4 promoter. AB - We have previously shown that E1A-mediated induction of the adenovirus E2 transcription unit likely involves the posttranslational activation of a previously limiting cellular factor termed E2F. However, this factor is not involved in E1A induction of several other viral genes, including the E4 gene, since it does not bind to the promoters of these genes. We have undertaken an analysis of proteins which bind to the E4 promoter in an attempt to define the basis for E1A control of this gene. Gel retardation binding assays revealed a large number of interactions with the E4 promoter consistent with the fact that at least 180 nucleotides of sequence are required for full promoter activity. The analysis was simplified by employing small probes as well as by using partially fractionated extracts. By so doing, we have identified at least seven discrete factor interactions involving the E4 promotor. Multiple interactions, as defined by discrete gel complexes, were identified with a site previously shown to be critical for promoter activity as well as E1A control. We find that one of these factors, termed E4F, is increased at least 10-fold in extracts prepared from Ad5 infected cells and that the increase requires the E1A gene. Furthermore, the activation is maximal by 3 h post-infection, consistent with the kinetics of activation of E4 transcription. Competition binding assays demonstrated that the E4F factor was E4 specific and did not interact with any other E1A inducible promoter. We therefore conclude that the induced E4F factor is likely responsible for the E1A-induced transcription of E4, thereby suggesting that E1A control must involve an activation of multiple promoter specific binding proteins. PMID- 2965008 TI - Increase in atrial natriuretic peptide in response to physical exercise. AB - Circulating atrial natriuretic peptide (ANP) level was determined during physical exercise to investigate the correlation between changes in ANP level and heart rate increases. Six subjects exercised at a work level of 75% VO2max for 30 min, two also performed two successive exercises at 75% VO2max while two more exercised for longer at 55% VO2max. Plasma ANP levels and heart rate increased in all the exercising subjects. At the end of the exercise, the ANP level fell immediately, suggesting an immediate reduction in ANP secretion by the heart. Pre exercise values were reached after 30 min. Successive exercises gave the same heart rate related ANP patterns without previous secretory episodes having any effect. These results lead to the conclusion that ANP intervenes in the cardiovascular adjustments to exercise. PMID- 2965009 TI - The responses of the catecholamines and beta-endorphin to brief maximal exercise in man. AB - The responses to brief maximal exercise of 10 male subjects have been studied. During 30 s of exercise on a non-motorized treadmill, the mean power output (mean +/- SD) was 424.8 +/- 41.9 W, peak power 653.3 +/- 103.0 W and the distance covered was 167.3 +/- 9.7 m. In response to the exercise blood lactate concentrations increased from 0.60 +/- 0.26 to 13.46 +/- 1.71 mmol.l-1 (p less than 0.001) and blood glucose concentrations from 4.25 +/- 0.45 to 5.59 +/- 0.67 mmol.l-1 (p less than 0.001). The severe nature of the exercise is indicated by the fall in blood pH from 7.38 +/- 0.02 to 7.16 +/- 0.07 (p less than 0.001) and the estimated decrease in plasma volume of 11.5 +/- 3.4% (p less than 0.001). The plasma catecholamine concentrations increased from 2.2 +/- 0.6 to 13.4 +/- 6.4 nmol.l-1 (p less than 0.001) and 0.2 +/- 0.2 to 1.4 +/- 0.6 nmol.l-1 (p less than 0.001) for noradrenaline (NA) and adrenaline (AD) respectively. The plasma concentration of the opioid beta-endorphin increased in response to the exercise from less than 5.0 to 10.2 +/- 3.9 p mol.l-1. The post-exercise AD concentrations correlated with those for lactate as well as with changes in pH and the decrease in plasma volume. Post-exercise beta-endorphin levels correlated with the peak speed attained during the sprint and the subjects peak power to weight ratio. These results suggest that the increases in plasma adrenaline are related to those factors that reflect the stress of the exercise and the contribution of anaerobic metabolism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965010 TI - The interaction of troponin-I with the N-terminal region of actin. AB - The interaction between troponin-I and actin that underlies thin-filament regulation in striated muscle has been studied using proton magnetic resonance spectroscopy. A restricted portion of skeletal muscle troponin-I (residues 96 116) has previously been shown to be capable of inhibiting the MgATPase activity of actomyosin in a manner enhanced by tropomyosin [Syska et al. (1976) Biochem. J. 153, 375-387]. On the basis of homologous spectral effects for signals of specific groups observed in different complexes formed using the native proteins and a variety of defined peptides, it is concluded that the segment of troponin-I which has inhibitory activity interacts with the N-terminal region of actin. The surface of contact of the inhibitory segment of troponin-I with actin involves two regions of the N-terminal of actin. These are located between residues 1-7 and 19-44. The data are discussed in the context of a structural mechanism for the inhibition of myosin ATPase activation. PMID- 2965011 TI - Purification of the beta subunit of the fibronectin receptor. AB - In this work we describe a method for purification of the beta subunit of the mouse fibronectin receptor (GP135). Cellular glycoproteins were isolated from a detergent extract of SR-Balb tumor cell membranes by two steps of affinity chromatography on lentil lectin-Sepharose and wheat-germ-agglutinin--agarose. This material was subsequently bound to an Affi gel 102 column and eluted by increasing salt concentration. Most of the GP135 was eluted at 80 mM sodium chloride together with a few other components. A final step of hydroxyapatite chromatography in sodium dodecyl sulphate allowed elution of GP135 as a single chromatographic peak. Fractions containing GP135 were identified at each chromatographic step by immunoblotting with a specific antiserum. By this procedure GP135 was purified to homogeneity as judged by SDS-PAGE analysis of 125I-labelled material. PMID- 2965012 TI - The role of MgATP hydrolysis in nitrogenase catalysis. AB - Kinetic studies on MgATP hydrolysis by nitrogenase of Azotobacter vinelandii were performed in the presence and in the absence of reducing equivalents. By measuring the ATPase activity of dye-oxidized nitrogenase proteins it can be excluded that reductant-independent ATPase activity is the result of futile cycling of electrons. The turnover rates of MoFe protein during reductant dependent and reductant-independent ATPase activity, when measured with excess Fe protein, have approximately the same value, i.e. 5 s-1 at pH 7.4 and 22 degrees C, assuming the hydrolysis of four molecules of MgATP per turnover of MoFe protein. For Fe protein on the other hand, the maximum turnover rate during reductant-independent ATPase activity is only about 6% of that of reductant dependent ATPase activity. While the reductant-dependent ATPase activity shows a sigmoidal dependence on the concentration of MgATP, the reductant-independent ATPase activity yields hyperbolic saturation curves. To account for these results it is proposed that the rate-limiting step during MgATP hydrolysis by oxidized nitrogenase is the rate of regeneration of active Fe protein. In the presence of reductant, the regeneration of active Fe protein is stimulated, explaining the higher ATPase activity of nitrogenase during substrate reduction. PMID- 2965013 TI - Left ventricular function during exercise before and after percutaneous transluminal coronary angioplasty using intravenous digital subtraction angiocardiography. PMID- 2965014 TI - Personal answer to a personal view on balloon aortic valvuloplasty. PMID- 2965015 TI - Inhibition of rat mammary carcinogenesis by an arotinoid without a polar end group (Ro 15-0778). AB - The influence of an arotinoid without a polar end group (Ro 15-0778) on rat mammary carcinogenesis was investigated. Mammary tumors were induced by oral administration of 15 mg, 7,12-dimethylbenz-(a) anthracene (DMBA) to 50-day-old female Sprague-Dawley rats. Ro 15-0778 inhibited the development of mammary adenocarcinomas. The percentage of tumor-bearing rats, the mean number of tumors per rat as well as the mean total volume of tumors per rat were dose-dependently reduced by Ro 15-0778. The results are of particular interest, since this compound--probably because of the lack of a polar end group--does not induce the signs and symptoms of hypervitaminosis A. The inhibition of mammary cancer development by Ro 15-0778 compares favorably with that of N-(4-hydroxyphenyl) retinamide the hitherto most effective retinoid for prevention of chemically induced mammary cancers in rats. PMID- 2965016 TI - Cell-populations involved in the humoral leukocyte adherence inhibition reaction. The T8-suppressor/cytotoxic cells are both effectors and responders. AB - In an attempt to identify the cell subpopulation involved in the humoral leukocyte adherence inhibition (H-LAI) reaction, mononuclear leukocytes were fractionated and the response with the different cell populations was measured in the H-LAI assay using sera from lung cancer patients and a lung cancer associated antigen. The monocytes were found to exhibit a suppressive effect on the H-LAI response. The B-lymphocytes seemed not to take an active part in the reaction. Depletion of the T-lymphocytes abolished the reaction. Subsequent studies on the T-lymphocyte subpopulations revealed that removal of the T4-cells had little effect while removal of the T8-cells resulted in complete abrogation of the H-LAI response. Direct blocking by monoclonal antibodies against the T4 and T8 surface determinants gave additional support for the T8-cell dependency of the H-LAI assay. The cells that specifically were induced to loose adherence in the H-LAI assay upon incubation with serum from a lung cancer patient and lung cancer associated antigen were also analyzed. Compared to the total mononuclear leukocyte population, the relative content of monocytes and B-cells were lower in the non-adherent population. An increase was found in the relative number of T cells. While the relative number of T4-cells decreased, the content of T8-cells was significantly increased. Furthermore, a direct correlation was found between the loss of adherence of the T8-subpopulation and the index of the H-LAI response. The present study indicates the T8-subpopulation of the T-lymphocytes to be involved both in the effector and responder side of the H-LAI reaction and gives implications for an immunological reaction behind the response of the H-LAI assay. PMID- 2965018 TI - Diagnostic differentiation between reactive and malignant lymphoid cells in serous effusions. AB - Lymphoid cell components in a total of 34 pleural and ascitic aspirates were investigated immunologically. The results indicate that reactive lymphocytes predominate in effusions from non-haemopoietic malignancies and benign conditions, while a significant proportion of fluids from patients with non Hodgkins lymphoma show unequivocal evidence of lymphomatous involvement. Immunological typing of lymphocytes in serous effusions is a valuable adjunct to conventional methods of diagnosis particularly in those patients in whom invasive procedures are undesirable. PMID- 2965017 TI - Proenzyme to urokinase-type plasminogen activator in human colon cancer: in vitro inhibition by monocyte minactivin after proteolytic activation. AB - Marked increases of plasminogen activator activity were observed in human colon cancer tissue, compared to corresponding normal tissues. This increase was attributable to urokinase-type activator (HPA52), with no increase evident in the level of the tissue-type plasminogen activator (HPA66). Human monocyte minactivin specifically inhibited HPA52 activity in cancer tissue homogenates and in colon cancer cell supernatants, an effect that was greatly enhanced by preincubation with plasminogen, indicating that the predominant form of HPA52 in tissue and the form that is secreted in vitro is the proenzyme. Inactivation of HPA52 by minactivin was shown to be dependent on proteolytic activation of HPA52 proenzyme. Utilization of HPA52 activity by tumors in vivo could therefore be dependent upon a protease, such as plasmin, to generate the extracellular proteolytic activity necessary to digest the intercellular matrix and permit invasion of normal tissue structures by colon cancer cells. PMID- 2965019 TI - Lack of effect of 5-aminosalicylic acid on platelet aggregation and fibrinolytic activity in vivo and in vitro. AB - We have studied platelet aggregation and fibrinolytic activity in six patients with chronic inflammatory bowel disease treated with 5-aminosalicylic acid (mesalazine). There were no changes in these measurements during normal treatment, i.e. 1.5 g per day with a slow-release formulation, nor after an intravenous dose of 250 mg. Also in vitro tests were negative, in contrast to the inhibition seen with aspirin (acetylsalicylic acid). We conclude that treatment with mesalazine does not constitute a hazard to these patients in regard to prolonged bleeding time caused by an influence on platelet aggregation or fibrinolytic activity. PMID- 2965020 TI - cDNA cloning of murine interleukin-HP1: homology with human interleukin 6. AB - Interleukin-HP1 (HP1) is a murine T cell-derived lymphokine, originally described as a growth factor for B cell hybridomas and plasmacytomas, that was recently shown to stimulate growth and differentiation of normal B and T lymphocytes. Here, we describe a cDNA for HP1 that was isolated from a library prepared using mRNA of a murine helper T cell clone activated with a clonotypic antibody. The cDNA, which hybridizes with a mRNA of approximately 1300 bp, encodes a polypeptide consisting of 211 amino acids with a typical signal sequence of 24 residues followed by 187 amino acids, which form the mature protein (Mr = 21,710). No N-glycosylation site but several potential O-glycosylation sites were identified in the predicted sequence. Comparison of the cDNA sequence of HP1 with that of human interleukin 6 disclosed a homology of 65% at the DNA level and of 42% at the protein level with a maximum of 57% for the segment spanning residues 42-102 of mature HP1. Considering the functional homology that was previously established between these two proteins, we therefore propose that HP1 be renamed murine interleukin 6. PMID- 2965021 TI - Stimulation with dendritic cells decreases or obviates the CD4+ helper cell requirement in cytotoxic T lymphocyte responses. AB - We investigated the need for CD4+ helper T (Th) cells in the induction of murine cytotoxic T lymphocyte (Tc) responses across minor or major histocompatibility (MHC) antigenic differences with either normal spleen cells (NSC) or purified dendritic cells (DC) as antigen-presenting cells (APC). Generation of a secondary in vitro class II MHC-specific Tc response was totally CD4+ Th cell-dependent with both types of APC. Likewise, male antigen (H-Y)-primed class II mutant bm12 T cells, which do not respond to H-Y presented on NSC, do respond to H-Y presented on DC in a completely CD4+ Th cell-dependent fashion. All other Tc responses, including primary anti-class I MHC, primary anti-class I + II MHC plus anti-minor H, and secondary C57BL/6 (B6) anti-H-Y, although not completely CD4+ Th cell dependent, were greatly augmented in the presence of CD4+ Th cells, but only with NSC as APC. In contrast, with DC as APC these responses were entirely or largely CD4+ Th cell independent. Similarly, H-Y primed class I MHC mutant bm14 T cells, which do not respond to H-Y presented on NSC, do respond to H-Y presented on DC in a completely CD4+ Th cell-independent fashion. The combined results indicate that DC can directly present class I MHC alloantigen or class I MHC plus nominal antigen (e.g. minor H) to CD8+ cells and generate a Tc response by these cells without the requirement for CD4+ Th cells. PMID- 2965022 TI - Quantitative and qualitative analysis of the Fc receptor for IgE (Fc epsilon RII) on human eosinophils. AB - In order to characterize the Fc receptor for IgE (Fc epsilon RII) on human eosinophils, we have compared the binding of human IgE myeloma protein to that of a monoclonal antibody (mAb BB10) directed against a common antigenic determinant of the Fc epsilon RII present on eosinophils, platelets and macrophages. Scatchard analysis of the binding to human eosinophils of the BB10 mAb revealed a linear monophasic binding curve, with a binding affinity of 1.17 x 10(7) M-1 and a number of 10(5) binding sites per cell. Biochemical analysis of the human eosinophil Fc epsilon R, performed by immunosorbent chromatography with either BB10 mAb or IgE, showed under nonreducing conditions a major component of 200 kDa. Under reducing conditions, 3 peptide fragments were obtained, with molecular masses of 45-50, 23 and 15 kDa. Finally, comparative analysis suggested that the Fc epsilon RII of human eosinophils and of a human macrophage cell line (U937) are structurally related and differ from the high-affinity Fc epsilon RI present on basophilic granulocytes. PMID- 2965023 TI - Induction of an immune response to a self antigen. AB - The question has been addressed whether the endogenous B cell population of a mouse can be induced to secrete antibodies specific for a self antigen present in serum. The antigen studied was the fifth component of mouse complement (C5). Nude BALB/c mice which are C5 sufficient were used as a source of potentially C5 reactive B cells and endogenous serum C5 provided the antigenic stimulus. We purposely avoided immunization with C5 in adjuvant. T cells from C5-deficient mice which lack this component in serum and are therefore not tolerant of C5 were injected into nude mice as a source of T cell help for anti-C5 reactive B cells. Control groups received T cells from C5-sufficient euthymic donors, which are tolerant of C5. Initiation of a response to C5 was monitored by testing the hemolytic function of serum. Reduction of C5-dependent hemolysis was observed in sera of mice which had received T cells from C5-deficient donors. Recipients of T cells from C5-sufficient donors maintained normal hemolytic complement levels throughout the test period of 45 days. Reduction of functional complement levels correlated with the presence of immune complexes of anti-C5/C5. C5-specific antibodies were mainly IgG1 and carried the IgG1 allotype of BALB/c providing unequivocal evidence that they were derived from the endogenous B cell population of the C5-sufficient host. PMID- 2965025 TI - Serotonin-glutamate interaction in rat cerebellum: involvement of 5-HT1 and 5-HT2 receptors. AB - The effects of serotonin (5-HT) on the release of endogenous glutamate (GLU) in rat cerebellum were investigated in slices depolarized with 35 mM K+. The Ca2+ dependent release of GLU was potently inhibited by 5-HT in a concentration dependent way. Release was also inhibited by the 5-HT1 receptor agonist 8-hydroxy 2-(di-n-propylamino)tetralin (8-OH-DPAT) and by the 5-HT2 receptor agonist 1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane HCl (DOI). The inhibition by 10 nM 5-HT was partly (35-40%) counteracted by the 5-HT2 receptor antagonist ketanserin but was fully blocked by the mixed 5-HT1/5-HT2 receptor antagonist methiothepin. The effect of 8-OH-DPAT was not affected by ketanserin but was totally antagonized by methiothepin, while the effect of DOI was entirely suppressed by ketanserin. Ketanserin or methiothepin themselves increased (by 23 and 55%, respectively, at 10 nM) the K+-evoked release of GLU. In conclusion the release of endogenous GLU in rat cerebellum can be inhibited by 5-HT through receptors of the 5-HT1 and 5 HT2 type. The enhancement of GLU release by ketanserin or methiothepin could suggest a tonic inhibition. The possible localization of the 5-HT receptors involved in the interaction with the GLU systems is discussed. PMID- 2965024 TI - Sequence and organization of the human T cell delta chain gene. AB - A novel human T cell receptor (TcR) gene, located 85 kbp upstream to the C alpha coding regions, was isolated using human genomic clones to identify cDNA homologous to messages encoded by this region. The deduced protein sequence of this gene is highly homologous to that of the newly identified constant region found in the murine TcR alpha chain locus. This gene undergoes rearrangements and is expressed at the RNA level in human thymocytes, peripheral T cells and several leukemic T cell lines which have been shown to express the surface gamma-delta heterodimer, suggesting that this gene encodes the human T cell delta chain. PMID- 2965026 TI - A pharmacological analysis of the rat spinal cord serotonin (5-HT) autoreceptor. AB - The pharmacological characteristics of the presynaptic 5-HT receptor associated with the modulation of 5-HT release were investigated in a preparation of rat spinal cord synaptosomes (nerve terminals) superfused with a Tris-buffered Krebs solution containing fluoxetine (1 microM). The 5-HT receptor agonists serotonin (1-100 nM), lysergic acid diethylamide (10 nM-1 microM) and the 5-HT1B receptor agonists 1-(m-trifluoromethylphenyl)piperazine (100 nM-1 microM) and 1-(m chlorophenyl) piperazine (100 nM-3 microM) concentration dependently decreased [3H]5-HT release, while 8-hydroxy-2-(di-n-propylamino)tetralin, a selective 5 HT1A receptor agonist, was inactive. The actions of the effective agonists were reversed by quipazine, an antagonist with high affinity for 5-HT1B binding sites, but not by spiperone, a 5-HT1A receptor antagonist. Furthermore, mesulergine, a 5 HT1C receptor antagonist was ineffective in reversing the action of 5-HT on [3H]5 HT release. These data indicate that the rat spinal cord nerve terminal autoreceptor has characteristics similar to the 5-HT1B binding site. PMID- 2965027 TI - Effect of niacin/nicotinamide deficiency on the diabetogenic effect of streptozotocin. AB - Weanling CD-1 mice were fed either a control diet or a diet deficient in niacin/nicotinamide for one month and then injected i.v. with 60, 80, 100, 120, 140, or 160 mg/kg streptozotocin. Mice on the deficient diet developed a higher incidence of diabetes and more severe hyperglycemia than those on the control diet. PMID- 2965028 TI - Ventricular fibrillation threshold during acute ischemia in hypertrophied rat hearts. AB - Ventricular fibrillation threshold was significantly lower in hypertrophied hearts than in normal hearts. Ischemia produced by coronary occlusion reduced fibrillation threshold in both normal and hypertrophied hearts, but the maximum reduction in fibrillation threshold was observed earlier in hypertrophied hearts. PMID- 2965029 TI - Effect of dexamethasone on Fc gamma receptor expression in foetal and neonatal rat gut. AB - When injected into 12-day-old suckling rats, dexamethasone caused a precocious disappearance of Fc gamma receptors from enterocytes of the proximal small intestine. However, dexamethasone appeared to be necessary for the maintenance or production of such receptors in foetal rat gut cultured in vitro. PMID- 2965030 TI - Collagen metabolism in the myocardium of normal and diabetic rats. AB - Myocardial collagen and total protein synthesis were studied in normal, diabetic, and insulin-treated diabetic rats after a single intraperitoneal injection of L [2,3-3H]proline as a radioisotopic precursor. The incorporation of tritiated proline into myocardial protein was regarded as a measure of total protein synthesis and the incorporation into hydroxyproline as indicative of myocardial collagen synthesis. Both total protein and collagen synthesis were found to be significantly lower in diabetic rats. This was associated with decreased degradation of both total protein and collagen in diabetic rats, as suggested by prolonged turnover times. Collagen content was also found to be increased in diabetic myocardium. Early insulin therapy with normalization of blood sugars in diabetic rats returned myocardial collagen metabolism to normal. This suggests that maintenance of euglycemia in diabetic rats is necessary to prevent abnormal myocardial collagen metabolism. PMID- 2965032 TI - Histologic evaluation of the luteal phase in women following follicle aspiration for oocyte retrieval. AB - The luteal phase was evaluated by endometrial biopsy (EMB) 11 to 13 days after administration of human chorionic gonadotropin (hCG) or initiation of the luteinizing hormone (LH) surge (hCG/LH) in 25 stimulated patients not undergoing embryo transfer (ET). Luteal phase defect (LPD) was defined as an EMB greater than 2 days out-of-phase when compared with the onset of subsequent menses. Nineteen women (76%) had LPDs ranging from 3 to 7 days. Mean estradiol (E2) on day of hCG/LH was significantly (P less than 0.01) higher (1709 +/- 997) in patients with in-phase EMBs than mean E2 levels in two groups of patients with out-of-phase EMBs (687 +/- 251, 941 +/- 366, respectively). These findings are further evidence of the importance of optimal follicular development to subsequent endometrial maturation. PMID- 2965031 TI - The migratory behavior of avian embryonic cells does not require phosphorylation of the fibronectin-receptor complex. AB - When locomotory embryonic cells become stationary, they acquire new substratum adhesion properties. In particular, the distribution of fibronectin receptors shifts from diffuse and highly mobile on the cell membrane to immobilized in close association with fibronectin molecules and cytoskeletal elements in focal contacts. Receptor phosphorylation has been proposed as a possible regulator of the interaction between the receptor and its intracellular and extracellular ligands. In the present study, we have compared the phosphorylation state of the fibronectin receptor in motile neural crest and somitic cells, in stationary somitic cells, and in Rous-sarcoma virus transformed-chick embryo fibroblasts, using immunoprecipitation following metabolic labeling. While no receptor phosphorylation was detected in motile embryonic cells, the beta subunit of the receptor was phosphorylated in stationary cells. This subunit was also highly phosphorylated in Rous-sarcoma virus-transformed chicken cells. These results suggest that phosphorylation of the fibronectin receptor cannot account for its distribution in the cell membrane and for the nature of the interactions between this receptor and its ligands in embryonic cells. PMID- 2965033 TI - Insulin-like growth factor-I (IGF-I) and IGF-I receptor in human testis: an immunohistochemical study. AB - Using specific monoclonal antibodies, the authors studied the distribution of insulin growth factor-I (IGF-I) and its receptor in normal human testis by immunostaining techniques. IGF-I was preferentially localized in Sertoli cells. Less evident positivity was found in primary spermatocytes. In the interstitium some Leydig cells were positive for IGF-I. The major positivity for the IGF-I receptor was found in secondary spermatocytes and early spermatids, whereas Sertoli cells were less positive. An intense positivity was found in some Leydig cells. PMID- 2965034 TI - Management of multiple conceptions after gonadotropin-releasing hormone analog/human menopausal gonadotropin/human chorionic gonadotropin therapy. AB - The treatment course of a 31-year-old infertility patient due to PCO disease is presented. Because the patient failed to conceive after various treatment cycles with CC, she was subjected to a combined GnRHa/hMG/hCG therapy. After plasma E2 levels had reached 2400 pg/ml, three leading follicles, with diameters of 20 to 24 mm, were detected. Induction of ovulation was achieved by 10,000 IU hCG. The patient conceived and developed ovarian hyperstimulation. At 8 weeks of gestation, seven cystic structures were detected within the uterine cavity, five containing single embryos, and two with twin embryos. All nine embryos were vital, as evidenced by their heart beats. Embryo reduction was achieved by transabdominal puncture on three occasions. The three surviving fetuses were carried to the 34th week of gestation. After delivery by cesarean section, three healthy babies developed normally. This communication illustrates the complications that can be associated with ovulation induction in PCO disease: ovarian hyperstimulation, polyovulation, multiple conceptions, and their clinical management. PMID- 2965035 TI - [Effects of estrogen and endocrine therapeutic agents on the estrogen receptor, progesterone receptor and DNA synthesis in MCF-7 human breast cancer cells using the whole cell uptake method]. AB - In order to investigate the mechanisms of the endocrine therapeutic agents and the applicability of combined endocrine therapies for breast cancers, we studied the effects of estrogen and the endocrine therapeutic agents on estrogen receptor (ER), progesterone receptor (PgR), and DNA synthesis in MCF-7 human breast cancer cells, which is known to be sensitive to estrogen. ER and PgR of MCF-7 cells were determined by whole cell uptake method. In brief, intact MCF-7 cells cultured in the multi-well plates were incubated with various concentrations of tritiated estradiol (E2) or promegestone (R5020) at 37 degrees C for 1 hour. The characteristics of the hormone binding were analyzed by Scatchard plots. MCF-7 cells had single class of ER (Kd: 2.1 +/- 0.2 X 10(-10) M, MBC: 9.0 +/- 1.5 X 10(3) sites/cell) and PgR (Kd: 7.2 +/- 1.1 X 10(-10) M, MBC: 3.1 +/- 0.5 X 10(4) sites/cell). When cultured in the presence of 10(-8) M or 10(-6) M of E2, tamoxifen (TAM), R5020 or medroxyprogesterone acetate (MPA) for 48 hrs, the numbers of binding sites of ER and PgR altered, but the affinities of either of them did not change. E2(10(-8) or 10(-6) M) increased about twice the number of PgR. Although treatment of 10(-8) M TAM, a non-steroidal antiestrogen, slightly increased the number of PgR, 10(-6) M TAM significantly decreased the number of PgR. Both of R5020 (10(-8) or 10(-6) M) and MPA (10(-8) or 10(-6) M), synthetic progestins, decreased the number of ER dose-dependently. On the other hand, E2 (10(-8) and 10(-6) M) and R5020 (10(-8) M) enhanced DNA synthesis, but 10(-6) M TAM or MPA inhibited DNA synthesis. The effects of single, sequential and coincidental treatment of these agents were compared. The sequential treatment of 10(-8) M E2 for 36 hrs and followed 10(-6) M MPA for 36 hrs ("10(-8) M E2----10( 6) M MPA") and "10(-6) M TAM----10(-6) M MPA" inhibited DNA synthesis of MCF-7 cells more efficiently than 10(-6) M TAM alone or 10(-6) M MPA alone for 72 hrs. However, "10(-6) M MPA----10(-6) M TAM" inhibited DNA synthesis less than "10(-6) M TAM----10(-6) M MPA".(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2965036 TI - Lymphocyte subpopulations in patients with atopic dermatitis. PMID- 2965037 TI - Suppressive characteristics of the cultured umbilical cord blood lymphocytes: enhanced suppression of non specific MLR by short term cultured peripheral blood and rosetted lymphocytes. AB - We have used the mixed leukocyte reaction assay (MLR) to compare the non-specific suppressor activity of fresh and short-term cultured peripheral blood rosetted lymphocytes from human umbilical cord blood. To a one way MLR between adult responders and irradiated adult stimulator cells, we added adult or cord blood mononuclear cells. Cord-blood derived mononuclear cells were more potent suppressors of the MLR than the adult peripheral blood mononuclear cells. When the fresh cord blood T cell fraction was separated from non-T cells by rosetting, the frequency and capacity of suppression fell, suggesting a role of non-T cells in this phenomenon. However, short-term culture (up to nine days) of cord blood derived T cells in the absence of mitogen resulted in a significant increase in their suppressive characteristics. Maternal cells were not used in any of these assays and the suppression was non specific. There was no HLA restriction to the response. The cord blood lymphocytes responded poorly to alloantigen when compared to the adult lymphocytes. In addition, there was a correlation between the allogeneic proliferative response and the suppressive capacity of the cord blood cells. Such a correlation was not found with the adult peripheral blood lymphocytes. PMID- 2965038 TI - Increased concentration of circulating atrial natriuretic peptide during normal pregnancy. AB - Atrial natriuretic peptide (ANP) is a recently discovered cardiac hormone involved in blood-volume homeostasis. Known stimulating factors for ANP release are rise in atrial pressures or atrial distension, suggesting that blood volume regulates ANP release. This study was undertaken to test the hypothesis that plasma levels of ANP are high and increase during normal pregnancy secondary to the expanding plasma volume. In a cross-sectional study plasma concentrations of ANP were measured in 99 normal pregnant women at different gestational ages and compared with the values found in an age-matched non-pregnant control group. Mean plasma ANP was already significantly increased in the first trimester as opposed to the non-pregnant women, but despite a continuously expanded plasma volume there was no further increase during pregnancy. Our findings suggest that other factors must interact with plasma volume in regulating plasma ANP during pregnancy. PMID- 2965039 TI - Variability of PI calculations. AB - In obstetrics the pulsatility index (PI) may be a useful parameter for the determination of fetal well-being. Although the PI is unambiguously defined, calculation from the measured data can be difficult. In order to study the comparability of PI values found by several investigators and with different methods tape recordings of Doppler signals of fetal vessels were used. The tapes were processed by several centres in Europe and the results were compared. Comparability is poor, although for the umbilical artery the results are in reasonable agreement. The method used has its drawbacks but shows that one must be careful in comparing results from different centres. PMID- 2965040 TI - Characterization of ATPase activity in the mycelial form cells of yeast Saccharomycopsis fibuligera. AB - 1. The properties of ATPase activity were studied with the mycelial form cells of Saccharomycopsis fibuligera. 2. Optimal pH for the activity was about 9.5. 3. The activity was stimulated by Mg2+. 4. The activity was inhibited by DCCD, NaF and oligomycin, but not inhibited by ouabain. PMID- 2965041 TI - Distribution of the flux control in convergent metabolic pathways: theory and application to experimental and simulated systems. AB - 1. Metabolic systems involving branched convergent pathways are analyzed under Flux Control Theory, obtaining a relationship between the contribution of every convergent pathway to the total flux and its Flux Control Coefficient. 2. An experimental model system is carried out to demonstrate the physical application of some conclusions of theoretical treatment. 3. Two different types of branched pathways are simulated by computer. 4. In both cases results are in agreement with the theoretical conclusions, showing in addition some new aspects on metabolic control. PMID- 2965042 TI - Biochemical effects of gossypol in isolated mitochondria: monovalent cations and ATPase activity. AB - 1. The effect of gossypol acetic acid upon the energy-producing system of isolated liver mitochondria was studied. Gossypol was found to exert an uncoupling effect. 2. Oxygen consumption and ATPase activity were stimulated only when a monovalent alkali metal cations was present. 3. The mitochondrial proton gradient, produced by the enzymatic hydrolysis of ATP, was lower in the gossypol treated mitochondria than in the control experiments. 4. It is proposed that gossypol stimulates oxygen consumption and the ATPase activity in mitochondria by a protonophoretic effect and a cation translocation process. PMID- 2965043 TI - The pharmacokinetics of loratadine in normal geriatric volunteers. AB - The pharmacokinetics of loratadine, a non-sedating anti-histamine, were studied in 12 normal geriatric volunteers. In an open label fashion, each volunteer received one 40 mg loratadine capsule. Blood was collected prior to and at specified times (up to 120 h) after dosing. Plasma loratadine concentrations were determined by a specific radioimmunoassay and those of an active metabolite, descarboethoxyloratadine, by high performance liquid chromatography. Concentrations of loratadine in the disposition phase were fitted to a biexponential equation and those of descarboethoxyloratadine to either a monoexponential or biexponential equation for pharmacokinetic analysis. Loratadine was rapidly absorbed, reaching a maximum plasma concentration of 50.5 ng/ml at 1.5 h after dosing. The disposition half-lives of loratadine in the distribution and elimination phases were 1.5 and 18.2 h, respectively. The area under the plasma concentration-time curve, was 146.7 h.ng/ml. Descarboethoxyloratadine had a maximum plasma concentration of 28.0 ng/ml at 2.9 h post-dose and an area under the concentration-time curve of 394.9 h.ng/ml. Its disposition half-lives in the distribution and elimination phases were 2.8 and 17.4 h, respectively. Comparison of these data with those from a previous study of loratadine in young adults showed no clear differences in the disposition half lives between the two groups. The clearance of loratadine tends to be lower in the elderly, but inter-individual variation within each age group appears greater than any age effect. PMID- 2965044 TI - Tolerance induced by grafting semi-allogeneic adult skin to larval Xenopus laevis: possible involvement of specific suppressor cell activity. AB - Major histocompatibility complex (MHC)-homozygous Xenopus laevis were rendered tolerant to semi-allogeneic antigens by grafting skins of adult frogs during larval stages (larvally induced tolerance), and this tolerant state was compared with the tolerance induced in early thymectomized frogs by the grafting of semi allogeneic nonlymphoid thymuses (thymus-reconstituted tolerance). In contrast to a total inability of thymus-reconstituted frogs both to reject skins and to exhibit a mixed leukocyte reaction (MLR) against the semi-allogeneic donor, larvally induced tolerant frogs showed a strong MLR against leukocytes of the tolerizing skin donor (split tolerance). Breakdown of the tolerant state in thymus-reconstituted frogs were easily accomplished by inoculation with syngeneic splenocytes, but this breakdown was extremely difficult to achieve in frogs with larvally induced tolerance. The injection of splenocytes from larvally induced tolerant frogs into normal frogs significantly suppressed semi-allogeneic graft rejection in the latter group; no suppression was obtained when splenocytes from thymus-reconstituted frogs were used. In addition, in the thymectomized frogs, recovery of allograft rejection capacity against the pertinent semi-allogeneic antigens were suppressed by the injection of splenocytes from larvally induced tolerant frogs, with the degree of suppression depending on the splenocyte dose. These results indicate that the larvally induced tolerant state is maintained by specifically induced suppressor cells affecting the in vivo allograft response but not the MLR. PMID- 2965045 TI - [Age-related evolution of the basic indicators of hemodynamics in schoolchildren and adolescents living in a mountain climate]. PMID- 2965046 TI - [Effect of excessive stimulation of the nervous system on aldosterone receptors in renal hypertrophy and insufficient stimulation on the interaction of aldosterone with renal tissue in reflex dystrophy]. PMID- 2965047 TI - Human immunodeficiency virus-induced pathology favored by cellular transmission and activation. AB - Epidemiological data suggest that transmission of human immunodeficiency virus (HIV) occurs primarily by transference of virally infected cells. However, the efficiency of lytic productive infection induced by HIV after transmission of cell-associated virus vs. free virus is difficult to assess. The present studies compare the extent of depletion of CD4+ (helper/inducer) T cells after mixing uninfected cells with either free HIV or irradiated HIV-infected allogeneic or autologous cells in vitro. Rapid CD4+ cellular depletion occurred only in cultures containing allogeneic infected cells or after addition of a nonspecific T cell activation signal to cultures with autologous infected cells. These in vitro observations strongly support the epidemiological implication that interactions between infected and uninfected cells are the most efficient means of transmission and HIV-induced cytopathology in vivo. They also provide direct support for the concept that immunological stimulation by foreign cells infected with HIV dramatically increases the likelihood of transmission. These in vitro observations suggest a model for the acquisition of HIV in vivo and the role of cellular activation in dissemination of the virus to uninfected cells in an infected individual. PMID- 2965049 TI - Experience with diagnostic laparoscopy in a hepatology training program. AB - Laparoscopy is an underutilized endoscopic procedure in the United States even though it has proven to be an important diagnostic modality. Among factors that may account for the unpopularity of this procedure are lack of experience and proper training in many gastroenterology fellowship programs, a perceived notion that there is a relatively high complication rate, and the preference for radiologic imaging techniques for establishing a diagnosis. We reviewed, over an 11-month period, our experience with laparoscopy in a large teaching center to better assess its safety when performed by trainees. The major and minor complication rates were 0.9% and 2.9%, respectively, with no fatalities. These findings compare favorably with the general experience. PMID- 2965048 TI - Measurement of portal blood flow by an ultrasonic "duplex" system composed of a pulsed Doppler flowmeter and a linear-type real time B mode electroscanner. AB - The portal vein hemodynamics of patients with various liver diseases were investigated by means of a duplex system consisting of a linear electroscanner and a pulsed Doppler flowmeter. In cases of chronic liver disease, the cross sectional area of the portal vein trunk became greater as the liver injury proceeded, while the maximum velocity tended to decrease. However, blood flow volume was no different between the controls and the chronic liver disease group. On the other hand, the splenic venous flow volume tended to increase as the liver injury advanced, suggesting that the increase in splenic venous flow volume is closely related to the formation of esophageal varices. This method permits non invasive observation of changes during the course of various types of liver disease. PMID- 2965050 TI - Laparoscopic cholecystostomy and the double bubble catheter. PMID- 2965051 TI - [Acute dermal gangrene: a postoperative complication. With an example from obstetrics]. AB - In the anglo-american literature the acute dermal gangrene is characterized as a cutaneous syndrome of infectious etiology which is usually leading- postoperatively or after an injury--to necrotizing destructions of the cutis, subcutis, subcutaneous fat, fascia or muscle. Depending on the localization of the involved tissue a superficial progressive bacterial gangrene is distinguished from a more deeply localized necrotizing fasciitis. Diagnostic and therapeutic considerations of these extremely rare syndromes are described in connection with an obstetric case which has been seen by us recently. PMID- 2965052 TI - [Immunological imbalance in carriers of the B8 antigen of the HLA system]. PMID- 2965053 TI - Different peripheral vasodilator effects of isradipine in sodium-loaded and sodium-depleted rabbits. AB - 1. The influence of acute changes in salt balance on the cardiovascular effects of the calcium antagonist isradipine were investigated in two groups of 6 rabbits pretreated 24 hr before an acute experiment, one by substituting 0.45% NaCl for drinking water, the other one with a subcutaneous injection of 20 mg kg-1 furosemide. 2. On the next day the animals were anaesthetized with barbiturates and 10 ml kg-1 0.9% NaCl or 5% glucose were infused into the salt loaded and salt depleted animals respectively. 3. Regional blood flows were then measured with microspheres before and after i.v. administration of 3 and 10 micrograms kg-1 isradipine (PN200-110). 4. Salt depleted animals had a lower cardiac output and lower flows to several regional vascular beds including the kidneys. 5. The isradipine effects on systemic haemodynamic variables were not different in the two groups. 6. However, the effects on regional blood flows were modified. In salt loaded animals skeletal muscle flow increased more, starting from a higher baseline, yet flow to the kidneys, pancrease and spleen was higher before and decreased significantly more after the calcium antagonist. 7. The results suggest that an alteration in the sodium balance selectively affects the mechanism responsible for vascular tone in a few vascular beds. PMID- 2965054 TI - Musculoskeletal pain among general dentists. PMID- 2965055 TI - Hereditary hemorrhagic telangiectasia. PMID- 2965056 TI - The role of the general dentist in the treatment and management of oral complications of chemotherapy. PMID- 2965057 TI - Prevention of bacterial endocarditis in dental patients. PMID- 2965058 TI - Dental emergency in a HIV-positive patient. PMID- 2965059 TI - Molecular cloning of cDNA for human prostatic acid phosphatase. AB - A human liver cDNA library in lambda gt11 was screened with polyclonal antiserum to human acid phosphatase isoenzyme 2a/4. About eleven positive clones have been obtained. Two clones, lambda Hap21 and lambda Hap22 were further characterized: clone lambda Hap21 contained a 0.8-kb cDNA insert and clone lambda Hap22 a 1.8 2.0-kb insert. XbaI digestion of lambda Hap22 generated two fragments of 1.0 and 0.9 kb. BglII digestion resulted in a 1.2-kb fragment and several smaller fragments of undetermined size. Clone lambda Hap22 contained all the genes carried by lambda gt11(lac5cI857nin5Sam100) and the 2-kb insert. An Escherichia coli(lambda Hap22) lysogen was generated, and its acid phosphatase activity was approximately ten-fold higher than that in the control nonlysogenic lysate. Western-blot analysis of total proteins present in this E. coli(lambda Hap22) lysate revealed that the non-induced lambda Hap22 prophage directed the synthesis of an approx. 175-kDa protein. This protein was recognized by antibody to the human acid phosphatase isoenzyme 2a/4 and anti-beta-galactosidase and was produced only upon induction with IPTG. These results indicated that lambda Hap22 carried a major portion of the gene coding for the human acid phosphatase isoenzyme 2a and/or 4 and this protein fragment of acid phosphatase was sufficient to manifest enzymatic activity. PMID- 2965060 TI - A 'phase-shift' fusion system for the regulation of foreign gene expression by lambda repressor in gram-negative bacteria. AB - A 'phase-shift' translation fusion vector was constructed in which mutually compatible restriction sites BamHI, BclI and BglII are positioned in such a manner that the cut point is in a different reading frame, immediately following the ATG start codon and ribosome-binding site of the lambda cro gene. The lambda cro gene is expressed from promoter pR and controlled by a thermosensitive (cI857) lambda repressor. The usefulness of the expression vector was demonstrated using a galK gene lacking the ATG start codon and fusing this to the pR promoter and ATG start codon of the lambda cro gene, resulting in cI857 regulated expression of galactokinase. The vector is of general use for foreign gene expression in Escherichia coli when the target gene has a compatible cohesive end (5'-GATC-3') at the N terminus (provided, for example, by a BamHI linker). The lambda cI857-pR-cro-galK cassette was cloned into pJRD215, a wide host-range plasmid and transferred by conjugation to a variety of Gram-negative bacteria. In all cases, thermosensitive regulation of galactokinase could be demonstrated, though the levels of induction varied considerably. These results show that the powerful lambda pR promoter and the efficient lambda repressor can be used to regulate expression of foreign genes in Gram-negative organisms other than E. coli. PMID- 2965061 TI - The overproduction of DNA terminase of coliphage lambda. AB - An artificial operon containing the genes coding for the two subunits of lambda DNA terminase, Nul and A, has been constructed. Derivatives of plasmid pBR322 served as the cloning vehicles. The transcription is driven by the pL promoter of phage lambda, and translation of the terminase genes was made efficient by the replacement of the wild-type ribosome-binding sites for those of lambda genes cII and/or D. The operon also carries the oL operator, and this enables regulation of its expression by a thermosensitive repressor. The synthesis of genes Nul and A products is extremely efficient upon derepression. Within 40 min after induction of the operon, the two subunits comprise about 20% of the total cellular protein mass. Crude extracts prepared from these overproducing strains are at least 100 times more active than extracts prepared from induced lambda lysogens in both promotion of lambda DNA packaging and cosmid cleaving. The ability to produce highly concentrated terminase would enormously facilitate the study of its structure and mechanism of action. These extracts are also extremely useful in techniques such as lambda DNA packaging, cosmid mapping and cosmid linearization to improve efficiency of integration into mouse eggs. PMID- 2965062 TI - High-level expression of alpha-human atrial natriuretic peptide from multiple joined genes in Escherichia coli. AB - A method is described which allows alpha-human atrial natriuretic peptide to be synthesized in stable form and with high yield in Escherichia coli. In the final expression system, eight copies of the synthetic alpha-hANP gene were linked in tandem, separated by codons specifying a 4-amino-acid (aa) linker with lysine residues flanking the authentic N and C termini of the 28-aa hormone. This sequence was in turn joined to the 3' end of a fragment containing the lac promoter and a leader sequence coding for the first seven N-terminal amino acids of beta-galactosidase. The expressed multidomain protein accumulated intracellularly into stable inclusion bodies and was easily purified by urea extraction of the insoluble cell fraction. The purified protein was cleaved into monomers by digestion with endoproteinase Lys-C, trimmed to expose the authentic C terminus by digestion with carboxypeptidase-B and a single disulfide bond was formed by gentle oxidation with potassium ferricyanide. The fully processed recombinant peptide was shown by reverse phase liquid chromatography to be indistinguishable from the chemically synthesized standard alpha-hANP in both the reduced and in the folded form. PMID- 2965063 TI - Functional elements of DNA upstream from the integrase operon that are conserved in bacteriophages 434 and lambda. AB - A 1488-bp restriction fragment of bacteriophage 434 DNA contains the integrase promoter and an adjacent nucleotide sequence (t'I) resembling a Rho-independent terminator. To identify and quantitate transcription termination, DNA segments were cloned into a plasmid between the galactose promoter and assayable galactokinase gene and tested for termination. Whereas the entire fragment effectively terminated transcription, a 331-bp restriction fragment containing the t'I terminator had only weak terminator activity. Random sequential deletions of the 434 DNA segment defined a strong terminator 650-bp upstream from t'I. This proposed Rho-independent terminator called tL4 consists of a 7-bp stem and 6-nt loop followed by a uridine-rich region in the RNA. Phage lambda contains an even stronger tL4 terminator that differs in 4 nt from 434 tL4. Thus, despite some sequence divergence, terminator activity has been conserved in these phages. The 434 DNA segment was also tested for promoter activity. Rightward promoter activity (opposite to pL in the phage) was located about 200 bp to the right of tL4 and was followed by an open reading frame (ORF) capable of encoding a 91 amino acid protein. Promoter activity in the same approximate location was also found in phage lambda. Thus the rightward promoter, the tL4 and t'I terminators, and ORF-55 all are elements in this segment of the genome that are conserved for function despite sequence divergence. PMID- 2965065 TI - [The Leningrad Sanitary and Hygienic Medical Institute and problems of reorganization of higher medical education]. PMID- 2965064 TI - Silent myocardial ischemia in the elderly: current concepts. AB - The traditional approach to managing ischemic heart disease has been to reduce symptoms, eg, angina pectoris. However, recent studies demonstrate that myocardial ischemia often can be found in the absence of common symptoms, including in elderly patients. Since elderly patients may not complain about their symptoms, or may not get enough exercise to test their cardiovascular capabilities and, thus, bring out problems, the geriatrician needs to be highly suspicious of silent myocardial ischemia. A brief overview of diagnostic testing methods suitable for the elderly and medical and surgical management options is presented. PMID- 2965066 TI - [Medico-epidemiological services in the Vologda region during World War II (1941 1945)]. PMID- 2965067 TI - [Current complex work with regard to conducting sanitary control of food-handling departments at industrial plants]. PMID- 2965068 TI - [Experience with the activities of the student sanitary-epidemiological division of the medical institute]. PMID- 2965069 TI - [Experience with activities with regard to improving the forms of sanitary control of maternity hospitals]. PMID- 2965070 TI - [Daily routine and health status of schoolchildren in extended-day groups]. PMID- 2965071 TI - [Working conditions and hygienic measures at flour mills]. PMID- 2965072 TI - [The role of daily hygienic treatment of working clothes among the measures for preventing pyoderma in coal miners]. PMID- 2965073 TI - [Functional status of the cardiovascular system of schoolchildren in the preparatory grades in different types of organization of the recreation periods]. PMID- 2965074 TI - [Various physiological indicators in vocational school students]. PMID- 2965077 TI - [Maternal age in pregnancy--genetic counseling and prenatal diagnosis in the light of known facts and personal observations]. PMID- 2965076 TI - [Assessment of the efficacy of supplementary vitaminization of workers engaged in underground repair of oil wells in the Azerbaijan SSR]. PMID- 2965075 TI - [Experience gained in the prevention of vibration disease in the Gorki region]. PMID- 2965078 TI - The autonomic nervous system and fatigue. AB - Behavioral, physiologic and exertional fatigue is differently defined, though symptoms are similar. The beneficial effect of amantadine on fatiguability in multiple sclerosis is accompanied by neuropeptide and lactate changes in the circulation. Exercise sometimes overwhelms temperature regulating mechanisms and may be associated with heat stroke. Endogenous opioids are markedly increased in the circulation during heat stroke and the use of specific opioid antagonists therapeutically has been proposed for heat stroke. Sympathetic activity changes in endurance trained subjects and vasoconstrictor responses are markedly attenuated. Similar changes occur in parasympathetic function which can be abnormal in up to 90% of endurance trained subjects. Hormonal secretion during prolonged exertion is altered and the normal signals (inhibiting or activating feedback mechanisms) are different in endurance trained subjects. Altitude, associated with acute mountain sickness, is also accompanied by an increase in cranial bloodflow. Circadian and temporal variation in autonomic function are manifest by changes in mast cell numbers and 5-HT containing nerve fibers in temple skin of patients with cluster headache. The remission rate induced by vagal stimulation in subjects with intractable hiccups is also affected by circadian hormonal or neurogenic influences. PMID- 2965080 TI - Overview of sleep disorders and the implication on dental practice. PMID- 2965079 TI - Sympathetic activation and muscle spindle. AB - An adrenergic central control on fusimotor activity has been demonstrated since 1954. At peripheral level, a muscle spindle autonomic innervation is not clearly demonstrated in man, but several data seem to suggest a direct autonomic innervation of muscle spindle in cats and rabbits. Data obtained in patients with peripheral adrenergic denervation (Idiopathic Orthostatic Hypotension) suggest a prevalent control of central adrenergic pathways on muscle spindle sensitivity. PMID- 2965081 TI - Hexosaminidase I indicates maturation disarrangement in acute leukemias. PMID- 2965082 TI - Monoclonal-antibody-purged autologous bone marrow transplantation for relapsed non-T-cell acute lymphoblastic leukemia in childhood. PMID- 2965083 TI - Neonatal myeloproliferative disorders in Down's syndrome and congenital leukemias. PMID- 2965084 TI - Properties of the mouse mos proto-oncogene locus. PMID- 2965085 TI - Effectiveness of intraarterial plasminogen application in combination with percutaneous transluminal angioplasty (PTA) or catheter assisted lysis (CL) in patients with chronic peripheral occlusive disease of the lower limbs (POL). AB - The accepted correct procedure for treating occlusive arterial diseases includes surgical disobstruction, CL as well as PTA. Combined non-surgical strategies are effective in about 60% of these patients. However, a high risk of rethrombosis despite from the prophylaxis with anticoagulants like heparin or antiplatelet drugs like ASA is proven, especially in patients with multi-segmental stenosis as well as in patients with extensive narrowing of the arteries. In these cases primary lesions (endangitis obliterans) or secondary lesions of the endothelium cause local depletion of plasminogen in the endothelium. Independent of the method used for reopening the vessel in these patients, a significant progression of the vessel disease and a high rethrombosis rate during longterm follow-up is observed. These results lead us to apply plasminogen locally to decrease the rate of rethrombosis. In patients suffering from stage III-IV (La Fontaine) including patients with multi-segmental stenosis as well as extended narrowing of the artery, PTA in combination with CL was performed. The catheter was placed as near as possible to the thrombus. In some cases the 'fibrinolyticum' could be injected directly into the thrombus. In these cases a bolus of 4,000 U/ml was locally infused, otherwise 1.0-1.5 million U urokinase per 24 hrs. were locally infused with heparin. In 28% (22 patients) no sufficient clinical response occurred using this combined therapy and plasminogen was applied locally. The following criteria supported our decision to include the patients in this study: 1. Insufficient response occurring after 12-24 hrs. of local infusion. 2. Following 6 bolus injections no reopening of the vessel occured within 60 minutes or the clinical response was insufficient due to rethrombosis. 3. Insufficient effects of lysis therapy after 2 hours and contraindication for a systemic fibrinolytic therapy (e.g. hypertension, age, etc.). 1,000 U plasminogen per ml were infused locally or 2,000 U up to 5,000 U plasminogen (in 5 to 10 ml 0.9% saline) were infused slowly (2-4 minutes infusion time) into the catheter in these patients 10 minutes after unsuccessful treatment with local urokinase therapy. Five minutes after administering plasminogen local intraarterial fibrinolytic therapy with urokinase was continued. No severe side effects due to this therapy were observed, although some patients suffered from acute pains in the peripheral segments of the arteries occurring immediately after infusion of plasminogen. In 16 of 22 patients a complete recanalization occurred and in 3 patients a satisfying clinical improvement was observed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2965087 TI - [Cardiac changes in athletes (athlete's heart)]. PMID- 2965086 TI - Congenital abnormal plasminogen, Frankfurt I, a cause for recurrent venous thrombosis. AB - A new abnormal plasminogen, Frankfurt I, has been identified in the plasma of a 42 year-old male patients who had recurring thromboses, thrombophlebitis and pulmonary embolism since his age of 29. Reduced functional and also slightly reduced antigen plasminogen concentrations were found in both the proposituts and his mother. Plasmin generation rates carried out by Streptokinase and Urokinase were also abnormal. The plasmin generated was very unstable in the absence of stabilizing ligands and/or substrates. Crossed immunoelectrophoresis of the purified Frankfurt I revealed a peak with normal size and shape, but displaced with respect to normal Glu-plasminogen toward the anode. Isoelectric focusing followed by zymography on an agarose-fibrin plate proved this observation but did not indicate a separation of the normal from the abnormal plasminogen molecular species, also, fewer bands were found in the abnormal plasminogen isozyme pattern. Kinetic studies of Frankfurt I Glu-plasminogen and plasmin showed that most of the functional abnormality is related to absence of active sites in half of the molecules. PMID- 2965088 TI - The anatomy of adipose tissue in captive Macaca monkeys and its implications for human biology. AB - In a sample of 31 sedentary, ad libitum-fed monkeys, most specimens had less than 5% adipose tissue by weight. Total fatness correlated closely with the number of adipocytes per kilogram lean body mass, but not at all with mean adipocyte volume, except in specimens below 5% fat. The total number of adipocytes per kilogram of lean body mass increased more than tenfold in the most obese specimens. These data suggest that, like humans but in contrast to laboratory rodents, adipocyte proliferation, not adipocyte enlargement, is the chief mechanism of adipose tissue expansion except in very lean monkeys. Adipose tissue was found in all the typical mammalian depots and in the superficial abdominal paunch, which enlarged disproportionately in obese specimens, forming an almost continuous layer over most of the body. Site-specific differences in the activities of some glycolytic enzymes were similar to those of other mammals. Adipocytes in the paunch depot showed biochemical properties in common with those in the groin depots. The distribution and cellularity of adipose tissue in normal humans were similar to those of exceptionally obese monkeys. Many of the interspecific and sex differences can be attributed to the much greater abundance of adipose tissue in humans, and may not be associated with hair reduction or aquatic habits. Some minor changes in the size or shape of certain adipose depots may have arisen recently under sexual selection. The relevance of laboratory rodents as animal models of human obesity is assessed from comparison of the cellular structure, anatomical distribution and enzyme profiles of adipose tissue in monkeys with those of human and other mammals. PMID- 2965089 TI - [Thrombolysis in acute infarct '87]. PMID- 2965090 TI - [Close to the edge of the pubic hair! Patient education and clinical skill responsibility in plastic surgery are legally evaluated differently]. PMID- 2965091 TI - A robot workstation for use in education of the physically handicapped. PMID- 2965092 TI - Using a robotic arm system to facilitate learning in very young disabled children. PMID- 2965093 TI - Suppressor T cells, immunoglobulin and Igh restriction. AB - Why do some T cell express idiotypes that are serologically similar to those of the B cells, since it is now well established that T cells do not use Ig genes for their antigen specific receptors? In this review article, we have summarized some the evidence for the influence of Igh linked genes on the Suppressor T cell repertoire, using anti-u treated mice as a model system. We investigated whether B cells and Ig molecules play a role in the generation of Ts repertoire. While our results clearly confirmed that B cells and Ig are important in the establishment of Ts repertoire, our experiments failed to resolve the fundamental question - 'what dictates the Igh restriction specificity of Ts?' Ts cells from anti-u treated mice did not lose all Igh restriction specificity. Instead, they expressed an altered restriction specificity. These results suggest that there are at least two independent mechanisms responsible for the generation of the suppressor T cell repertoire. A pre-selected germ line one, which is Ig independent, and a mature one, which is Ig dependent. The precise mechanisms responsible for the generation of germ line Ts repertoire remain to be determined. PMID- 2965094 TI - Structure, organization and polymorphism of murine and human T-cell receptor alpha and beta chain gene families. PMID- 2965095 TI - Induction of tolerance towards TNP entails down-regulation of an autoimmune attack. AB - In order to follow the process of induction and maintainance of tolerance, BALB/c mice were tolerized by free hapten, and effector and regulatory cell interactions were analysed by limiting-dilution (LD) cultures. Injection of trinitrobenzenesulphonic acid (TNBS) resulted, predominantly, in the activation and expansion of self-reactive cytotoxic T cells (CTL), which were observed transiently at frequencies comparable to allo-specific CTL. In addition, self reactive helper T cells (Th) were activated and expanded in tolerized mice. TNP specific reactivity was difficult to evaluate, since cytotoxic activity against haptenized self followed the pattern of self-reactivity throughout the test period. But in LD cultures determining proliferation, two populations of Th responding to TNP-self were observed, while only one Th population could be detected in response to self. Expansion/activation of Th and CTL precursors (CTLp) was followed by activation of suppressor T cells (Ts). The suppressor population could be divided into two subpopulations, one interfering with Th, the second interacting directly with CTL (veto cells). The results indicate that during the induction of tolerance, animals pass through an autoimmune attack, with expansion and activation of self-reactive clones (CTL, Th). The final status of non-responsiveness towards TNP is not due to the deletion of effector or regulatory cells, but results from the establishment of a steady state of dominance of self-reactive and TNP-self-reactive suppression. PMID- 2965096 TI - The effect of interferon-gamma treatment of rat tumour cells on their susceptibility to natural killer cell, macrophage and cytotoxic T-cell killing. AB - The effect of exposing tumour cells to interferon-gamma (IFN-gamma) has been studied investigating changes in MHC antigen expression and susceptibility to a variety of cellular effector mechanisms. Treatment of rat tumour cell lines with rat recombinant IFN-gamma increased their expression of class I MHC molecules, as monitored by quantitative immunofluorescence. Reduced sensitivity to lysis by NK cells was observed, although cold-target competition assays indicated NK cells were bound equally well by both interferon-treated and control cells. Decreased susceptibility to NK lysis was not a general effect of IFN-gamma treatment of tumour cells, as the sensitivity to lysis by activated macrophages was unaffected. Furthermore, both allogeneic CTL and CTL from syngeneic tumour-immune rats were able to kill IFN-gamma-treated tumour cells more effectively than untreated control target cells. These studies show that IFN changes the balance of sensitivity to different cytolytic effector mechanisms, increasing T-cell mediated effects, decreasing NK killing, but leaving macrophage cytotoxicity unaffected. The impact of these changes on tumour rejection will depend on the relative contribution of these mechanisms to target-cell destruction. PMID- 2965097 TI - Investigation of the mechanism of active enhancement of renal allograft survival by blood transfusion. AB - Transfusion with donor blood before transplantation can prolong the survival of renal allografts in the rat. The phenomenon is donor specific. We have investigated the effect of blood transfusion on both lymphocyte proliferation and the generation of donor-specific cytotoxic cells. In the first instance the kinetics of the proliferative responses of lymph node, spleen and thoracic duct lymph (TDL) leucocytes (harvested 7 days after transfusion of donor-specific blood, third-party allogeneic blood, syngeneic blood or no treatment) to lymph node stimulator cells were determined in a one-way mixed lymphocyte culture (MLC). Four strain combinations (LEW to DA, LEW to PVG, PVG to DA and DA to PVG) were investigated in this study. Maximal proliferation was observed at 4 days in the MLC and this was not altered by prior blood transfusion. When lymph node and TDL cells from transfused recipients were used as responders and stimulation was provided by cells from the same strain of rat as the blood donor, there was a significant depression of the MLC response compared to the response of cells harvested from untreated animals or animals given a syngeneic blood transfusion. This correlated with the behaviour of renal allografts in the same strain combinations. Lymph node and TDL cells harvested from rats that had received a third-party blood transfusion produced variable results in the MLC, ranging from a normal to a depressed response. No prolongation of allograft survival was noted in any animals pretreated with third-party blood. In contrast, when spleen cells were used as responder cells there was a donor-specific increase in the proliferative response on Day 4 in all strain combinations tested. In the second instance, specific cytotoxic activity was generated by in vitro (MLC) stimulation of lymph node or TDL, but not spleen, cell responders harvested from transfused or untreated rats. The activity of cytotoxic cells generated in MLC was quantified in a 51Cr-release assay, using Con A blast lymph node target cells prepared from animals of the same strain as the blood donor or a third-party strain. The specific cytotoxic activity generated was significantly increased when the responding cells were harvested from an animal that had received a donor specific transfusion and then been restimulated by the blood donor in MLC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2965098 TI - In vitro induction of a contrasuppressor immunoregulatory network by polyclonally activated T cells derived from murine Peyer's patches. AB - Much evidence suggests that Peyer's patch (PP) lymphocytes are capable of mounting both humoral and cell-mediated immune responses to luminal antigenic stimuli. To shed further light on T-T and T-B cell interactions in gut mucosal immune-associated processes, we studied in vitro the effects of a variety of PP derived concanavalin A (Con A)-activated immunoregulatory T-cell subsets on class specific immunoglobulin (Ig) production by lipopolysaccharide (LPS)-activated PP derived B cells. Particular attention was focused on induction of a contrasuppressor T-cell immunoregulatory network in the above in vitro system. Three types of immunoregulatory effector T cells, a helper T (Th) cell, a suppressor T (Ts) cell and a contrasuppressor T (Tcs) cell were developed and isolated. The results showed that B cell Ig production was under the regulation of these T cells, and the L3T4+ Lyt-2- T cell, which bound to Vicia villosa (VV), had a contrasuppressor effector function. In addition, a L3T4+ Lyt-2- VV- Ts inducer (Tsi) subset and a L3T4- Lyt-2+ VV- Ts effector subset also appeared to participate in the sequential development of the suppressor and, probably, contrasuppressor immunoregulatory networks, respectively. Thus, PP T cells are likely to execute their highly sophisticated immunoregulatory functions, not only in the helper and suppressor circuits but also in the contrasuppressor circuit in response to intraluminal non-specific stimuli. However, IgA isotype-specific Ig production appears to be controlled primarily by the isotype-specific helper circuit, not by the contrasuppressor circuit, in polyclonal LPS-stimulated gut mucosal immune responses. PMID- 2965099 TI - Monoclonal antibodies which react with bovine T-lymphocyte antigens and induce blastogenesis: tissue distribution and functional characteristics of the target antigens. AB - In this study we report on the tissue distribution and functional characteristics of bovine T-cell differentiation antigens recognized by the monoclonal antibodies (mAbs) IL-A26, IL-A27, and IL-A28. All three mAbs are able to stimulate proliferation of peripheral blood mononuclear cells (PBM) and inhibit proliferation of responder cells in mixed leucocyte cultures (MLC). MAbs IL-A27 and IL-A28 are believed to react with the same molecule, which is different to that recognized by IL-A26 as determined by a number of criteria. MAbs IL-A27 and IL-A28 inhibit binding of one another, but not of IL-A26. MAbs IL-A27 and IL-A28 react with 25% of thymocytes confined to the medulla, whereas IL-A26 reacts with approximately 80% of thymocytes, including medullary and cortical populations. MAbs IL-A27 and IL-A28 react with thymocytes which express BoT4 or BoT8 singularly, whereas IL-A26 reacts with all cells which express BoT4 or BoT8, either singularly or dually, in addition to all thymocytes which react with IL A27/28. Only IL-A26 inhibits spontaneous sheep erythrocyte (E)-rosette formation by bovine T cells. Based on tissue distribution and functional characteristics, IL-A26 is believed to recognize the bovine homologue of CD2, designated BoT2, whereas IL-A27/28 reacts with a mature T-cell antigen. Cells reactive with the mAbs constitute approximately 60% of bovine PBM. Using these mAbs in dual immunofluorescence analyses, at least three populations of bovine T cells are demonstrable in PBM. The majority of T cells are BoT4+ or BoT8+ and also react with IL-A26/27/28. A second small population of PBM is negative for BoT4 and BoT8 but is IL-A26/27/28+. A third population (less than 5%) is BoT4-/BoT8-/ILA27/28- but reacts with IL-A26. PMID- 2965100 TI - Antibody-mediated erythrolysis and erythrophagocytosis by human monocytes, macrophages and activated macrophages. Evidence for distinction between involvement of high-affinity and low-affinity receptors for IgG by using different erythroid target cells. AB - Antibody-dependent cellular cytotoxicity (ADCC) and Fc receptor-mediated phagocytosis were determined with human monocytes, monocyte-derived macrophages and activated macrophages, using rabbit IgG-covered sheep red blood cells (EAs) and anti-D-treated human erythrocytes (EAhu) as target cells. Monocyte and macrophage-mediated ADCC were distinguished by different kinetics, monocytes lysing either target more rapidly than macrophages. Macrophage activation by recombinant IFN-gamma (rIFN-gamma) led to a marked increase in ADCC activity against EAhu. This manifested in increased lysis of optimally sensitized target cells, in a sustained lysis of target cells carrying low antibody densities, and as an enhanced resistance to lysis inhibition by competing fluid-phase inhibition by competing fluid-phase IgG. All these effects were less striking when EAs were the target cells. Phagocytosis of EAs by rIFN-gamma-treated cells was strongly suppressed, regardless of the amount of antibody on the target cells, and susceptibility to inhibition by fluid-phase IgG was slightly increased. By comparison, phagocytosis of EAhu was depressed to a lesser degree, and susceptibility to inhibition by fluid-phase IgG was reduced when macrophages were rIFN-gamma treated. These and other experiments suggested that the functional triggering of monocytes and macrophages by EAs involved, at least in part, low affinity Fc receptors (FcR), whereas EAhu interacted with macrophages via high affinity FcR. It is shown elsewhere that rIFN-gamma treatment of macrophages increases the expression of high-affinity FcR, but not low-affinity FcR (Jungi, Lerch & Brcic, 1987). Differences in the rIFN-gamma-induced functional alterations assessed with EAhu or with EAs are interpreted therefore as being a consequence of differential involvement of high-affinity FcR and of low-affinity FcR in mediating an effector function. For monitoring rIFN-gamma-induced alterations in the effector capacity EAs are more appropriate targets since up regulation of high-affinity FcR has a smaller influence on the response to this type of target. Using metabolic inhibitors, ADCC could be dissociated from ingestion suggesting that ADCC is not a post-phagocytic event. PMID- 2965102 TI - [Results of an experimental study on skin tolerance and photobiological aspects of an anti-acne product for topical use]. PMID- 2965101 TI - The role of macrophages in LPS-induced lethality and tissue injury. AB - In the present study we investigated the role of mononuclear phagocytes in the pathogenesis of lipopolysaccharide (LPS)-induced lethality and tissue injury. Since hepatic and splenic macrophages are the primary sites of localization of i.v.-injected LPS, we selectively eliminated these macrophages using liposome encapsulated dichloromethylene diphosphonate (DMDP). After double DMDP-liposome treatment the phagocytic cells in the liver and spleen were completely eliminated, except for the macrophages in the white pulp of the spleen which were affected to a lesser extent by this treatment. An i.v. injection of LPS into DMDP and saline-pretreated mice showed that the latter animals exhibited febrile associated symptoms such as lethargy and ruffled fur, but that macrophage elimination abrogated these symptoms. Although after double saline- or DMDP pretreatment the LD50 appears to be 1 mg and 630 micrograms, respectively, the differences in lethality between both groups of mice were not statistically significant. Therefore, we concluded that hepatic and splenic macrophages are not necessary for LPS-induced lethality. The role of macrophages in LPS-induced local tissue damage was studied by comparing the histopathological changes in hepatic and splenic tissue between DMDP- and saline-pretreated mice. A sublethal dose of LPS induced similar hepatic lesions in macrophage-depleted and saline-pretreated mice, whereas the histopathological changes in the spleen were much more pronounced after DMDP-pretreatment. Particularly in the inner periarteriolar lymphocyte sheath (PALS) of these mice, the number of T cells was considerably reduced and extensive cellular necrosis could be found. These data strongly suggest that the local tissue damage resulting from LPS injection may not be due to its localization in mononuclear phagocytes but rather to interaction with other cell types. PMID- 2965103 TI - [Thiomersal: a frequent cause of sensitization]. PMID- 2965104 TI - The evasion of the immune response and immunological unresponsiveness: parasitic helminth infections. AB - A selective review is given of the mechanisms associated with the evasion of the immune response by parasitic helminths and immunological unresponsiveness as it applies to helminth infections. Immunosuppression caused by parasites leading to reduced responsiveness of lymphocytes to mitogens is discussed, and mechanisms that inhibit effector mechanisms at the parasite surface and polyclonal activation of lymphocyte populations are dealt with. Physiological immunosuppression associated with parturition and lactation and the immunological unresponsiveness of young ruminants are dealt with in detail. PMID- 2965106 TI - Lymphocyte population dynamics in experimental murine diabetes. AB - Hyperglycemia was induced in BALB/c mice with alloxan monohydrate. The spleen and lymph nodes of control vs. diabetic groups were analyzed for total cell content and the distribution of lymphocyte subpopulations when immunized 1) with DNFB, a contact allergen or 2) horse apoferritin, a protein antigen or 3) non-immunized at quiescence. The total cell contents of spleen and lymph nodes from diabetic groups, when stimulated with either antigen or at quiescence, were less than respective control groups. The distributions of B lymphocytes, T lymphocytes and suppressor T lymphocytes in the spleen and lymph nodes of diabetic mice at quiescence or during either immune response did not differ from controls. Diabetic mice produced more horse apoferritin-specific antibody in vivo than controls and lymphocytes from DNFB sensitized diabetic mice proliferated in vitro in response to DNBSO3. However, the proportion of proliferative lymphocytes in situ in diabetic DNFB-challenged mice was decreased in comparison to DNFB challenged controls. From these data we conclude that hyperglycemia in alloxan diabetic mice non-specifically impedes, but does not completely inhibit, the proliferation and growth of lymphocyte populations. PMID- 2965105 TI - Trauma peptide-mediated prostaglandin E2 biosynthesis: a potential mechanism for trauma-induced immunosuppression. AB - In vitro exposure of peripheral-blood-adherent mononuclear cells or amnion cells to nanomolar quantities of a trauma-associated immunosuppressive peptide resulted in an increased biosynthesis of prostaglandin E2 (PGE2). Trauma peptide enhanced prostaglandin E2 biosynthesis by as much as 425% compared to buffer controls. The addition of trauma peptide to mixed lymphocyte cultures significantly inhibited [3H]thymidine incorporation by human peripheral blood lymphocytes. Addition of indomethacin (an inhibitor of prostaglandin biosynthesis) to mixed lymphocyte cultures did not significantly abrogate the immunosuppressive activity of the peptide. These results indicate that suppression of T lymphocyte blastogenesis by trauma peptide is probably mediated by at least two mechanisms: (1) by increased PGE2 biosynthesis, induced by trauma peptide, and (2) through a non cyclooxygenase-mediated pathway. PMID- 2965107 TI - Analysis of causes of laparoscopic sterilization failure. PMID- 2965108 TI - Factors contributing to resistant hypertension. Cardiac considerations. AB - Medical treatment of hypertension has beneficial effects in the prevention of complications of the disease and in prolongation of survival. Nevertheless, the control of hypertension may sometimes be difficult. In this respect, the heart itself may interfere with blood pressure control because of increased cardiac output or because of cardiogenic reflexes that could conceivably not only initiate but also perpetuate peripheral vasoconstriction. The resultant persistence of hypertension may induce further cardiac changes that will in turn perpetuate the rise in blood pressure. PMID- 2965109 TI - Leukocytosis and low serum IgA in workers exposed to the epoxy compound, t-methyl 3-phenylglycidate. AB - Forty-nine out of 54 male workers engaged in the production of an epoxy compound, t-methyl-3-phenylglycidate, showed skin symptoms in varying degrees that may be due to the skin-irritative effect of the compound. The exposed workers were also shown to have subjective symptoms which may be related to the irritative property of the compound on surface tissue. Laboratory examinations on the blood obtained from the exposed workers showed significantly higher values of leukocyte concentration as compared with the non-exposed controls. This was chiefly caused by the increase of neutrophilic granulocytes and T-cell lymphocytes. Serum IgA levels of the exposed workers were shown to be significantly lower than those of the control group. Hemoglobin concentration, hematocrit value and red cell count of the exposed workers remained at the same level as those of the control subjects. Liver or kidney damage was not found in biochemical analyses on the sera of exposed workers. PMID- 2965110 TI - Erythrocyte CR1 receptors in patients with hypogammaglobulinaemia. AB - Erythrocyte CR1 receptors were measured in 16 patients with primary hypogammaglobulinaemia and compared with those in 14 normal controls. There was no significant difference in the number of receptors in the two populations. One patient with hypogammaglobulinaemia had a very low number of receptors and subsequently developed red cell aplasia. PMID- 2965111 TI - Evidence for in vitro regulation of IgE receptors on the human basophil membrane by their removal and reexpression: effects of Ca2+, Mg2+, some metabolic inhibitors and fetal calf serum. AB - Human basophils free of receptor-bound IgE and suspended in RPMI-1640 + 0.2% EDTA or in a buffered salt solution lacking Ca2+ and Mg2+ bound optimal quantities of 125I IgE. In media containing Ca2+ and Mg2+ such as RPMI-1640 alone or a buffered physiologic salt solution binding was reduced significantly in the first 60 min and remained essentially unchanged in the next 120 min. Scatchard analysis showed the reduction to be due both to loss of number and affinity of receptors. The presence of both 2-deoxy-d-glucose and 2,4-dinitrophenol usually prevented the reduction of binding in RPMI-1640. Suspension of cells in RPMI-1640 supplemented with fetal calf serum (FCS) demonstrated reduction in binding at 60 min characteristic of divalent-cation-containing media but this was followed by a rise in binding to approximately control levels in the next 120 min. This pattern of binding occurred in about 70% of the experiments. In most of the remaining experiments there was no difference from control binding in that there was no early decrease and no late rise in binding. This variability was not leukocyte donor-dependent. When cells were preloaded with 125I IgE and incubated in RPMI 1640 dissociation of IgE was more rapid than in media free of Ca2+ and Mg2+. The results suggest that membrane-bound IgE receptors on basophils may be shed by an energy and Ca2+ and Mg2+ requiring process and reexpressed dependent on a factor or factors present in FCS. PMID- 2965112 TI - Per-operative uncontrollable bleeding at polyester (Dacron) arterial prosthesis implantation. AB - An aortobifemoral knitted polyester graft was placed in a 58-year-old woman with aorto-iliac occlusive disease. Although the prosthesis was preclotted in the usual manner, it never became impervious to blood under normal intra-arterial pressure. Examination of the graft showed lipid infiltration not ordinarily seen. We recommend that when a knitted prosthesis fails to preclot a woven graft should be used instead. PMID- 2965113 TI - Phenytoin-induced gum hypertrophy improved by isotretinoin. PMID- 2965114 TI - Transformation and mutagenic potential of porphyrin photodynamic therapy in mammalian cells. AB - The transformation and mutagenic potential of porphyrin photodynamic therapy has been examined in mammalian cells. The mutagenic frequency in Chinese hamster cells at the Na+/K+ ATPase locus was measured by resistance to ouabain following treatment with either photodynamic therapy (PDT) or UV irradiation. The C3H 10T 1/2 mouse embryo cell system was used to document the transformation frequency following PDT, UV irradiation, gamma irradiation or exposure to 3 methylcholanthrene (MCA). Treatments with UV irradiation were effective in producing mutants resistant to ouabain, and treatments with UV irradiation, gamma irradiation and MCA generated transformants at frequencies comparable to those which are reported in the literature. However, PDT treatment conditions (which produced a full range of cytotoxicity) did not induce any mutagenic or transformation activity above background levels. PMID- 2965115 TI - Synergistic inhibition of anatid herpesvirus replication by acyclovir and phosphonocompounds. AB - Plaque reduction assays were used to evaluate the inhibitory effects of acyclovir (ACV), phosphonoacetate (PAA), and phosphonoformate (PFA) with a plaque-purified isolate of anatid herpesvirus (AHV-ppc3). A plaque assay employing a liquid overlay medium was developed to facilitate the drug inhibition studies. From dose response curves, the ID50 for PAA, PFA, and ACV were 20, 12, and 0.14 micrograms/ml, respectively. From data obtained from combination dose-response curves, dose isobolograms were prepared and used to determine the types of interactions between drug pairs. Whereas the interaction between PFA and PAA was additive, synergism occurred with ACV and either PAA or PFA. Drug-resistant mutants of AHV-ppc3 resistant to 8.0 micrograms/ml ACV, 250 micrograms/ml PAA, 180 micrograms/ml PFA or 6.0 micrograms/ml AHV, and 220 micrograms/ml PAA were isolated. PMID- 2965116 TI - Technetium-99m labeled renal function and imaging agents: II. Clinical evaluation of 99mTc MAG3 (99mTc mercaptoacetylglycylglycylglycine). AB - First clinical results indicated that among all potential 99mTc-complexes the N3S complex 99mTc MAG3 has the biokinetic properties most similar to o-I-hippurate (OIH). A simultaneous comparison of 99mTc MAG3 and 131I OIH was conducted in a series of 48 patients. Twelve out of 18 transplant patients received simultaneously both radiopharmaceuticals for follow-up scintigraphic studies during the early post-transplantation period. In all of these double tracer studies (n = 90) the renograms obtained with 99mTc MAG3 exhibited qualitatively the same curve-shapes, compared with OIH, in cases of acute graft rejection or acute tubular necrosis. In addition, both agents were administered simultaneously to 46 of these patients and the plasma clearance values were determined under steady state conditions. The clearance data of both radio-pharmaceuticals showed a very good correlation (r = 0.95; P less than or equal to 0.05). The 99mTc MAG3/131I OIH clearance ratio showed a mean value of 0.65. Due to the high plasma protein binding of 99mTc MAG3 it is assumed not to be filtered through the glomerular membrane. Therefore we introduce for the 99mTc MAG3 clearance the term "Tubular Extraction Rate (TER)". Due to the favourable physical characteristics of 99mTc, higher activities can be administered as compared to 131I OIH. This allows the simultaneous examination of the tubular function as well as the renal perfusion. Additionally the kidney morphology is imaged with much better resolution. It is concluded that from the clinical point of view 99mTc MAG3 is a favourable replacement for OIH not only for renal imaging but also for the quantitative determination of tubular function. The acceptance will depend on the quality and on the handling of a commercially available kit formulation. PMID- 2965118 TI - A comparison of effects of zearalenone and estradiol benzoate on reproductive function during the estrous cycle in gilts. AB - Effects of estradiol benzoate (EB) and zearalenone (Z) on luteal maintenance and plasma hormone concentrations were studied in 45 gilts. Gilts were allocated to receive either 20 mg Z, 2 mg EB or no treatment (C) on d 1 to 5 (T1), 6 to 10 (T2) or 11 to 15 (T3) of an estrous cycle (five per treatment). Onset of estrus was designated as d 0 of the estrous cycle. Zearalenone was added to the daily ration and EB was administered via an intramuscular injection. Blood samples were collected every 10 min over a 4-h period on the first 2 d prior to onset of treatment; the first, third and fifth days of treatment; and the first two and the fifth day after the end of the treatment periods. Gilts receiving EB and Z during T2 and T3 had longer (P less than .05) inter-estrous intervals than C gilts. The range in inter-estrous intervals for Z and EB treatments was 28 to 74 and 27 to 63 d, respectively. Mean plasma progesterone concentrations were elevated (P less than .05) during T2 and T3 in EB and Z-treated gilts when compared with C females. Estradiol benzoate treatment during T2 and T3 reduced (P less than .05) mean plasma luteinizing hormone (LH) concentrations more than C or Z treatments. Mean plasma concentrations of 13, 14-dihydro-, 15-keto prostaglandin F2 alpha (PGFM) during T3 were higher (P less than .05) in C and Z gilts on d 13 and 15 post-estrus when compared with EB gilts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965117 TI - MZ15, a monoclonal antibody recognizing keratan sulphate, stains chick tendon. AB - Biochemical, morphometric and electron histochemical methods have failed to demonstrate the presence in tendon of keratan sulphate, a common component of connective tissue proteoglycans. Using a monoclonal antibody specific for keratan sulphate, a positive localization of this molecule was made in the gastrocnemius tendon of stage 44 chicken embryos both at the light and electron microscopical levels. PMID- 2965119 TI - Microbiological properties of teicoplanin. AB - Teicoplanin is a glycopeptide antibiotic structurally related to vancomycin. Both agents achieve their effect by binding to acyl-D-alanyl-D-alanine in the bacterial cell wall. The spectrum of activity of teicoplanin, like that of vancomycin, is restricted to Gram-positive aerobic and anaerobic bacteria. However, the activity of the two agents is not identical: teicoplanin is generally more active than vancomycin against streptococci and Gram-positive anaerobes; the two agents exhibit similar activity against Staphylococcus aureus (including methicillin-resistant strains); and vancomycin is more active than teicoplanin against some strains of coagulase-negative staphylococci. Inoculum size influences the activity of teicoplanin, and variations in activity against some strains of coagulase-negative staphylococci have been observed with different culture media. Resistance to teicoplanin and vancomycin is difficult to induce under laboratory conditions and the small increment in resistance that may develop is lost when the organisms are subcultured in the absence of the drugs. PMID- 2965120 TI - Teicoplanin--clinical use and laboratory correlation. Based on papers presented at the 15th International Congress of Chemotherapy. Istanbul, Turkey, July 1987. PMID- 2965121 TI - Teicoplanin in empirical combined antibiotic therapy of bacteraemias in bone marrow transplant patients. AB - Teicoplanin in combination with amikacin and ceftazidime was used as empirical therapy in treating 46 febrile episodes in 34 consecutive patients undergoing allogenic bone marrow transplantation. All but four of these febrile episodes occurred in neutropenic patients and 50% of them proved to be bacteraemias (23/46). Cure was achieved in 90% of Gram-positive bacteraemias (18/20) and in six of them teicoplanin was the only antibiotic with activity in vitro against the infecting strain. All (3/3) Gram-negative bacteraemias were cured. Central venous catheter removal was required in five patients (three tunnel infections, one exit-site infection and one thrombophlebitis). Two failures occurred among Gram-positive bacteraemias and in one case the patient died of infection. Four instances of side effects were documented but only one was severe (hearing loss). PMID- 2965122 TI - Teicoplanin in combination therapy for febrile episodes in neutropenic and non neutropenic paediatric patients. AB - The aim of this study was to assess the efficacy and safety of teicoplanin in combination with other antimicrobial agents for therapy of severe suspected or proven Gram-positive infection in children and also to determine a dosage regimen for paediatric patients. Twenty children were given 23 courses of teicoplanin therapy for 11 septicaemias, one erysipelas, one cellulitis and 11 cases of fever of unknown origin. Eighteen of the 20 patients had severe underlying disease: one solid tumour, 15 acute lymphoblastic leukaemias, two acute myeloblastic leukaemias; 15 were neutropenic; 19 had a central line. Thirteen Gram-positive bacteria were isolated from the blood cultures in eleven patients. There were eight coagulase-negative staphylococci (CNS), (five methicillin-resistant) and four Staphylococcus aureus isolates. Teicoplanin was given as a 30 min infusion twice on the first day then once a day. The mean unit dose was 6 mg/kg for first eight patients. One clinical failure and lower serum concentrations than expected led us to increase the dosage to 10 mg/kg daily for the remaining patients. Tolerability remained excellent. It is concluded that antistaphylococcal treatment for febrile episodes in neutropenic patients can be satisfactorily provided by teicoplanin 10 mg/kg iv daily with a second loading dose on the first day. One injection a day is a convenient schedule in paediatrics. PMID- 2965123 TI - Teicoplanin in the treatment of skin and soft tissue infections. AB - Twenty-five patients with cellulitis or other soft tissue infections were treated with once daily teicoplanin. One patient was withdrawn from the study because of a protocol violation but 27 of the 29 assessable infections in the other 24 patients were clinically cured or improved. Eighteen of the 20 original Gram positive bacterial isolates (11 Staphylococcus aureus and nine beta-haemolytic streptococcus strains) were completely eradicated. No severe adverse reactions were seen but a rise in the plasma platelet count was noted in the majority of the patients during teicoplanin administration. PMID- 2965124 TI - Efficacy and safety of teicoplanin in gram-positive peritonitis in patients on peritoneal dialysis. AB - Twelve cases of peritonitis caused by Gram-positive bacteria in 11 dialysis patients were treated with teicoplanin. Treatment, which was continued for three weeks, consisted of the addition of teicoplanin to the dialysis fluid. Six patients who were febrile on admission were also given a single intravenous dose of 400 mg teicoplanin. For patients on continuous ambulatory peritoneal dialysis 20 mg teicoplanin per litre was added to each dialysis bag during the first week of treatment, to alternate bags during the second week, and only to the overnight dwell bag in the third week. For patients on intermittent peritoneal dialysis, 20 mg/l teicoplanin was added at each dialysis session. Resolution of peritonitis occurred in all patients within one to five days (mean 2.2); nine were discharged within this period and the patients continued to treat themselves at home. The other two patients were kept in hospital for reasons unconnected with the peritonitis. Nine patients have remained well at follow-up 2-13 months (mean 6.3) later. Two patients, both of whom had Staphylococcus aureus peritonitis, relapsed three months after the end of treatment. Mean serum teicoplanin concentrations were less than 10 mg/l, except in one patient who was re-treated when he relapsed. No adverse effects were recorded; one patient who developed a conductive hearing loss was found to have otitis media and obstruction due to wax. We conclude that teicoplanin is safe and effective in treating peritonitis in patients on peritoneal dialysis. PMID- 2965125 TI - Intraperitoneal teicoplanin in the treatment of peritonitis associated with continuous ambulatory peritoneal dialysis. AB - The efficacy of teicoplanin in the treatment of peritonitis in patients undergoing continuous ambulatory peritoneal dialysis (CAPD) was evaluated in a randomised comparison with vancomycin. The dosage regimen used was 50 mg of vancomycin or teicoplanin per 2 1 bag of dialysate for 48 h followed by 25 mg per bag for a further five days. Twelve episodes of peritonitis were studied. There was no significant difference in the bacteriological or clinical cure rates of either antibiotic. PMID- 2965126 TI - A comparison of teicoplanin and cefuroxime as prophylaxis for orthopaedic implant surgery: a preliminary report. AB - The relative merits of different antibiotic regimens for prophylaxis in orthopaedic implant surgery are difficult to evaluate because of the low frequency of infection. Factors other than infection prevention may influence choice. We have compared 400 mg teicoplanin given intravenously on induction of anaesthesia with three perioperative injections of cefuroxime, in 146 patients undergoing total hip or total knee replacement. These interim results suggest that cefuroxime selects for increased extraintestinal carriage of faecal streptococci and teicoplanin for Proteus species. There were no significant differences between the regimen in the acquisition of coagulase-negative staphylococci or Clostridium difficile, post-operative diarrhoea, wound healing or wound infection. Both regimens were equally safe. PMID- 2965127 TI - Bactericidal activity of teicoplanin in an in-vitro two-compartment kinetic model. AB - The bactericidal activity of teicoplanin was studied in a model system in which antibiotic concentration is continuously varied. A two-compartment open model was used to reproduce, in vitro serum pharmacokinetics of teicoplanin in humans during 24 h after iv administration. Staphylococcal and streptococcal strains of various species were tested. Teicoplanin showed rapid killing of Staphylococcus epidermidis and Staph. hominis. All strains of Staph. epidermidis were rapidly killed in spite of the high MIC values (4-16 mg/l). The killing effect against Staph. haemolyticus was less rapid and marked than against Staph. epidermidis, with strain to strain variation. A 99.9% kill was achieved for most strains within 22 h, but with other isolates regrowth occurred which was due to the presence of highly resistant cells in the original population. Other staphylococci (Staph. aureus, Staph. saprophyticus) and Streptococcus pyogenes, Str. bovis and Str. salivarius were reduced by 99.9% within 22 h. Bactericidal activity against other streptococci (Str. mutans, Str. agalactiae, Str. faecalis, Str. faecium) was less marked; against Str. sanguis teicoplanin was bacteriostatic. PMID- 2965128 TI - Pharmacokinetics of 14C-teicoplanin in healthy volunteers. AB - Five healthy male volunteers received over 60 sec a single intravenous injection of 400 mg of teicoplanin labelled with 41 microCi of 14C. Plasma and urine total radioactivity was measured up to 10 and 16 days, respectively. Teicoplanin was assayed in plasma and urine also by a microbiological method, with similar results. Pharmacokinetic parameters were estimated by model-independent analysis and the following mean values were obtained: elimination half-life 77 h; total body clearance 9.8 ml/h/kg; renal clearance 7.81 ml/h/kg; volume of distribution at steady-state 0.759 l/kg. Similar estimates were obtained by a compartmental analysis. A total of 80% of the administered dose was recovered in urine in 16 days; 2.7% of the dose was recovered in faeces collected for eight days after administration. The mean total recovery of the drug was 83 +/- 0.6%. The plasma and urine concentrations of teicoplanin observed after a single 400 mg iv dose exceeded the MIC for most pathogens for at least one day, and this suggests that a daily dosage regimen would be satisfactory for patients with normal renal function. PMID- 2965129 TI - Pharmacokinetics of a single intravenous dose of teicoplanin in subjects with various degrees of renal impairment. AB - The pharmacokinetic profile of teicoplanin after single 3 mg/kg doses in adults with different degrees of renal failure was reviewed. The disposition of teicoplanin was adequately described by a three-compartment open pharmacokinetic model and it appears to be primarily related to the degree of renal function. Teicoplanin total body clearance was less and the terminal elimination half-life progressively prolonged in association with renal failure, but the volume of distribution at steady-state did not change. Highly significant relationships between teicoplanin total body and renal clearance and creatinine clearance have been reported and serve as a basis for adjusting dosage in patients with renal failure. In patients on continuous ambulatory peritoneal dialysis teicoplanin, administered either intravenously or intraperitoneally, showed bidirectional exchange characteristics through the peritoneal membrane, although transfer from the systemic circulation to peritoneal fluid was consistently low. Haemodialysis made no significant contribution to total body clearance of teicoplanin. Guidelines for administration of teicoplanin in patients with renal failure are discussed. PMID- 2965130 TI - Pharmacokinetics of teicoplanin in the elderly. AB - The pharmacokinetic profile of teicoplanin was studied in 12 elderly patients with a moderate degree of renal impairment (mean creatinine clearance, 51.3 ml/h/kg before treatment), after a single 6 mg/kg iv dose. Pharmacokinetic parameters were estimated both by a three-compartment open pharmacokinetic model and by non-compartmental analysis; peak plasma levels, 15 min after administration, averaged 45 mg/l. The half-lives of two distributive phases were 0.39 and 7.3 h, respectively. The elimination half-life averaged 107 h, with similar estimates obtained from the three-compartment analysis and from urinary data. The volume of distribution from the central compartment was 0.09 l/kg while the volumes of distribution at steady state and during the elimination phase were 1.3 and 1.6 l/kg, respectively. The total teicoplanin clearance averaged 10.6 ml/h/kg, with renal clearance accounting for about 40% of the total. There was a linear correlation between teicoplanin total or renal clearance and endogenous creatinine clearance. The average total recovery of teicoplanin in urine over eight days was 28%. There were no local or systemic adverse reactions to teicoplanin. PMID- 2965131 TI - Pharmacokinetics of teicoplanin in children. AB - A single dose of 6 mg/kg teicoplanin was infused over 10 min in six children of mean age 7 years, and a single dose of 6 mg/kg was infused over 20 min in four neonates of mean age 8.5 days. Serum sampling was performed at 0 and 10 min and at 1, 4, 12 and 24 h and thereafter 24-hourly, up to ten days. Urine was collected for each 24 h throughout the study in children. Teicoplanin was assayed by high performance liquid chromatography. Tolerability of teicoplanin was excellent both in children and in neonates. For all the patients the serum concentrations of teicoplanin followed an open two-compartment model. Mean Cmax was 48.6 +/- 16.7 mg/l (10 min) in children, and 19.6 +/- 1.05 mg/l (20 min) in neonates, and mean t1/2 beta was 20.5 +/- 5.5 h and 30.3 +/- 6.3 h, respectively. On the basis of the results dosage recommendations for children and neonates are made. PMID- 2965132 TI - HPLC quantitation of the six main components of teicoplanin in biological fluids. AB - Teicoplanin, a new glycopeptide antibiotic belonging to the same class as vancomycin, is a mixture of six main components, designated A3, A2-1, A2-2, A2-3, A2-4 and A2-5. An assay method by higher performance liquid chromatography (HPLC) has been devised to separate and monitor each of these components in blood and urine. PMID- 2965133 TI - A multicentre open clinical trial of teicoplanin in infections caused by gram positive bacteria. AB - A multicentre open trial of teicoplanin in 81 centres in nine European countries included 1431 cases: 531 female, 900 male; mean age 49.4 years, range 1-93 years. These were hospitalized patients most of whom had infections caused by Staphylococcus aureus (816 isolates). Of a total of 1427 Gram-positive pathogens 280 (19.6%) were methicillin resistant. There were 536 skin and soft tissue infections, 263 septicaemias, 135 lower respiratory tract infections, 179 joint and bone infections and 83 endocarditides. More than a third of the infections were severe. Complicating medical factors were present in 69% of cases, including malignant disease in 14% and diabetes mellitus in 11%. Mean teicoplanin dose was 289 mg/day; mean duration of treatment was 14 days. A total of 471 patients received a high dose regimen, 400 mg teicoplanin daily for at least five days. Monotherapy with teicoplanin was used in 1037 cases and combination with other antibiotics in 394. Overall 91.7% of the 1333 evaluable cases were clinical cured or improved. The MIC of teicoplanin was less than or equal to 1 mg/l for 90% of Gram-positive isolates. Adverse events were reported in 189 cases (13.2%). The most common drug-related event was an allergic type skin reaction which occurred in 35 cases (2.4%). Transient hepatic dysfunction was reported in 28 patients (2.0%). PMID- 2965134 TI - Clinical efficacy and safety of teicoplanin. AB - This was an open efficacy and safety study in 310 hospitalized patients (187 male and 123 female) with infection by Gram-positive bacteria. The age range was 12-88 years (mean 49.8 years), and 80 of the patients were older than 65 years. Teicoplanin was given either by fast iv or by im injection, in most cases at a dose of 200 or 400 mg every 24 h with an initial dose of 400 mg. For both routes of administration the mean total daily dose was 286 +/- 5 mg/day. The mean duration of treatment was 8.2 days (range 1-37 days). The infections comprised 133 skin or soft tissue, 53 joint or bone, 40 urinary tract, 39 upper respiratory tract, 21 lower respiratory tract, nine septicaemia, four gastro-intestinal tract and 11 miscellaneous. All Gram-positive bacteria isolated were sensitive to teicoplanin. Teicoplanin was the sole antimicrobial agent given in 269 of the 310 patients. Clinical cure occurred in 79.7%, and improvement in 13.6%. Adverse events related to teicoplanin occurred in 7.7% of the patients. Most were minor and in only six patients was treatment discontinued because of adverse events (three allergy, two bronchospasm, one tremor). The laboratory values and audiometry indices in 67 patients showed no significant changes. Teicoplanin was considered an effective and well-tolerated treatment for Gram-positive infections in this study. PMID- 2965136 TI - Teicoplanin in infections caused by methicillin-resistant staphylococci. AB - Serious infections by methicillin-resistant staphylococci, 26 caused by Staphylococcus aureus and two by coagulase-negative staphylococci, were treated with teicoplanin 200-800 mg daily, in eight cases supplemented with another antibiotic. The overall rate of clinical cure or improvement was 75%, and that of bacterial eradication 67%. PMID- 2965135 TI - Teicoplanin compared with vancomycin in methicillin-resistant Staphylococcus aureus infections: preliminary results. AB - Twenty-one patients were included in an open randomized study comparing vancomycin 1 g bd with teicoplanin 400 mg daily in severe methicillin-resistant Staphylococcus aureus infections. The median duration of therapy was 15 days for vancomycin and 21 days for teicoplanin. Most patients presented with severe underlying conditions, such as major surgery (8), solid tumours (5), multiple trauma (3). The infections treated, included septicaemia, osteomyelitis, bronchopneumonia, cellulitis and acute pyelonephritis. Mean MICs of the strains were 0.39 mg/l for vancomycin and 0.195 mg/l for teicoplanin. Mean trough and peak serum concentrations of vancomycin were 14.3 +/- 5 mg/l and 34.3 +/- 13 mg/l, while the teicoplanin values were 7.5 +/- 4 mg/l and 17 +/- 7 mg/l. The cure rate was seven of 12 in the teicoplanin group and six of nine in the vancomycin group, with four and three cases, respectively, of improvement and one failure in the teicoplanin group. Transient renal impairment occurred in two cases with both regimens; superinfection and colonization in three patients and one patient, respectively, with both regimens. PMID- 2965137 TI - Teicoplanin in the treatment of infections caused by coagulase-negative staphylococci. AB - A total of 88 patients were treated with teicoplanin for infections caused by coagulase-negative staphylococci in open clinical studies in France and the UK. Teicoplanin was administered once daily, with a mean dose of 323 mg, for a mean duration of 16 days. Thirty-nine patients received teicoplanin alone while 49 received combination treatment. Clinical cure or improvement occurred in 79 of 82 evaluable cases (96.3%) and bacteriological elimination in 82 cases (95.1%). Treatment with teicoplanin alone was clinically successful in all cases, including 20 septicaemias, and the elimination rate was 93%. There was no apparent correlation between clinical and bacteriological outcomes and results of in-vitro testing. The variation in MICs between France and the UK was attributed to differences in the methods used. PMID- 2965138 TI - Inherited immunoregulatory dysfunction in extrinsic bronchial asthma. AB - We studied 119 members of 22 asthmatic multiplex families. Included were: 44 parents (seven were asthmatics), 48 asthmatics (23 were undergoing an attack at the time of sampling), and 27 normal siblings. The following investigations were carried out on all subjects: 1) detection of total T lymphocytes, helper cells, and suppressor cells, using monoclonal antibodies (OKT3, OKT4, and OKT8), 2) study of nonspecific T-lymphocyte blast transformation induced by PHA, and 3) HLA A, B, and DR antigen determination using the microcytotoxicity technique. The results were compared with normal ranges and data for a normal group and statistically and genetically analyzed. They indicate that: 1) the number of T cells was low in asymptomatic asthmatics and normal in asthmatics in attack; 2) there were fewer helper and normal suppressor cells (that is, a low H:S ratio) in asymptomatic asthmatics, and a normal amount of helper and suppressor cells (a normal H:S ratio) in those experiencing an attack; 3) there was a percentage of lymphocyte transformation in both groups of asthmatics; 4) whereas the T-helper cells increased, there was no change in the number of suppressor cells during an attack, which points to deficient function of suppressor cells; 5) the disorder is inherited and the gene controlling this dysfunction is HLA-linked and probably dominant. PMID- 2965139 TI - Prevention of exercise-induced bronchospasm in asthmatic children. Effect of aerosol and oral procaterol hydrochloride. AB - This double-blind crossover study compared the therapeutic value of bronchodilator procaterol hydrochloride at 5 micrograms per puff by aerosol inhalation using a metered-dose inhaler and at 25 micrograms by oral administration using tablets with respective inert placebos in preventing exercise-induced bronchospasm (EIB) in 10 children with bronchial asthma. The two active dosage formulations produced a significant EIB inhibitory effect compared with the placebos. In a comparison of the two active preparations, the aerosol was more efficacious than the tablet in improving all respiratory parameters examined, and statistical significance was obtained in FEV1, MMEF, PEFR, V50, and V25 5 minutes after exercise. Further, the aerosol was beneficial in all subjects, while the tablet was beneficial in seven of 10. PMID- 2965140 TI - Oral clenbuterol and procaterol. A double-blind comparison of bronchodilator effects in children with chronic asthma. AB - In a double-blind crossover study, clenbuterol at a dose of 0.75 micrograms/kg, procaterol at a dose of 1.5 micrograms/kg, and placebo, all administered orally, were compared for bronchodilation efficacy. Twelve children aged from 6 to 13 years, with moderate to severe asthma, participated in the trial. Pulmonary function, heart rate, blood pressure, and tremor were evaluated at 30, 60, 90, and 120 minutes and then hourly for 8 hours after administration. Both clenbuterol and procaterol induced a significant change over their baseline values for all the pulmonary function parameters considered. For clenbuterol, the same was also observed in comparison with placebo, while for procaterol this was true only for FEV1 and FEF25-75, while no difference resulted from FVC and PEF. Mild and transient tremor was the only side effect observed. Oral clenbuterol and procaterol were both demonstrated to be safe and effective. However, at the doses studied, clenbuterol had a significantly higher bronchodilator activity lasting up to 8 hours. PMID- 2965141 TI - Cloning of genes from mucoid Pseudomonas aeruginosa which control spontaneous conversion to the alginate production phenotype. AB - Strains of Pseudomonas aeruginosa causing chronic pulmonary infections in patients with cystic fibrosis are known to convert to a mucoid form in vivo characterized by the production of the exopolysaccharide alginate. The alginate production trait is not stable, and mucoid strains frequently convert back to the nonmucoid form in vitro. The DNA involved in these spontaneous alginate conversions, referred to as algS, was shown here to map near hisI and pru markers on the chromosome of strain FRD, an isolate from a cystic fibrosis patient. Although cloning algS+ by trans-complementation was not possible, a clone (pJF5) was isolated that caused algS mutants to convert to the Alg+ phenotype at detectable frequencies (approximately 0.1%) in vitro. Gene replacement with transposon-marked pJF5 followed by mapping studies showed that pJF5 contained DNA transducibly close to algS in the chromosome. Another clone was identified called pJF15 which did contain algS+ from mucoid P. aeruginosa. The plasmid-borne algS+ locus could not complement spontaneous algS mutations in trans, but its cis acting activity was readily observed after gene replacement with the algS mutant chromosome by using an adjacent transposon as the selectable marker. pJF15 also contained a trans-active gene called algT+ in addition to the cis-active gene algS+. The algT gene was localized on pJF15 by using deletion mapping and transposon mutagenesis. By using gene replacement, algT::Tn501 mutants of P. aeruginosa were constructed which were shown to be complemented in trans by pJF15. Both algS and algT were located on a DNA fragment approximately 3 kilobases in size. The algS gene may be a genetic switch which regulates the process of alginate conversion. PMID- 2965142 TI - Lysogenization of Escherichia coli him+, himA, and himD hosts by bacteriophage Mu. AB - The possible outcomes of infection of Escherichia coli by bacteriophage Mu include lytic growth, lysogen formation, nonlysogenic surviving cells, and perhaps simple killing of the host. The influence of various parameters, including host himA and himD mutations, on lysogeny and cell survival is described. Mu does not grow lytically in or kill him bacteria but can lysogenize such hosts. Mu c+ lysogenizes about 8% of him+ bacteria infected at low multiplicity at 37 degrees C. The frequency of lysogens per infected him+ cell diminishes with increasing multiplicity of infection or with increasing temperature over the range from 30 to 42 degrees C. In him bacteria, the Mu lysogenization frequency increases from about 7% at low multiplicity of infection to approach a maximum where most but not all cells are lysogens at high multiplicity of infection. Lysogenization of him hosts by an assay phage marked with antibiotic resistance is enhanced by infection with unmarked auxiliary phage. This helping effect is possible for at least 1 h, suggesting that Mu infection results in formation of a stable intermediate. Mu immunity is not required for lysogenization of him hosts. We argue that in him bacteria, all Mu genomes which integrate into the host chromosome form lysogens. PMID- 2965143 TI - Intermediates in bacteriophage Mu lysogenization of Escherichia coli him hosts. AB - Characterization of a putative intermediate in the Mu lysogenization pathway is possible in a variant Escherichia coli himD strain which exhibits greatly diminished lysogen formation. In this strain, most infecting Mu genomes form stable, transcribable, nonreplicating structures. Many of these genomes can be mobilized to form lysogens by a second Mu infection, which can be delayed by at least 100 min. This intermediate structure can be formed in the absence of Mu A or B function. We suggest that the inferred intermediate could be the previously reported protein-linked circular form of the Mu genome. Providing Mu B function from a plasmid enhances Mu lysogenization in this him strain, and the enhancement is much greater when both Mu A and B functions are provided. PMID- 2965144 TI - Chromosome-mediated iron uptake system in pathogenic strains of Vibrio anguillarum. AB - We describe in this work a new iron uptake system encoded by chromosomal genes in pathogenic strains of Vibrio anguillarum. This iron uptake system differs from the plasmid-encoded anguibactin-mediated system present in certain strains of V. anguillarum in several properties. The siderophore anguibactin is not utilized as an external siderophore, and although characteristic outer membrane proteins are synthesized under iron-limiting conditions, these are not related to the plasmid mediated outer membrane protein OM2 associated with ferric anguibactin transport. Furthermore, the siderophore produced by the plasmidless strains may be functionally related to enterobactin as demonstrated by bioassays with enterobactin-deficient mutants, although its behavior under various chemical treatments suggested major differences from that siderophore. Hybridization experiments suggested that the V. anguillarum chromosome-mediated iron uptake system is unrelated genetically to either the anguibactin or enterobactin associated iron assimilation systems. PMID- 2965145 TI - Effect of calcofluor white on chitin synthases from Saccharomyces cerevisiae. AB - The growths of Saccharomyces cerevisiae wild-type strain and another strain containing a disrupted structural gene for chitin synthase (chs1::URA3), defective in chitin synthase 1 (Chs1) but showing a new chitin synthase activity (Chs2), were affected by Calcofluor. To be effective, the interaction of Calcofluor with growing cells had to occur at around pH 6. Treatment of growing cells from these strains with the fluorochrome led to an increase in the total levels of Chs1 and Chs2 activities measured on permeabilized cells. During treatment, basal levels (activities expressed in the absence of exogenous proteolytic activation) of Chs1 and Chs2 increased nine- and fourfold, respectively, through a mechanism dependent on protein synthesis, since the effect was abolished by cycloheximide. During alpha-factor treatment, both Chs1 and Chs2 levels increased; however, as opposed to what occurred during the mitotic cell cycle, there was no further increase in Chs1 or Chs2 activities by Calcofluor treatment. PMID- 2965146 TI - Transforming growth factor beta stimulates the expression of fibronectin and of both subunits of the human fibronectin receptor by cultured human lung fibroblasts. AB - Transforming growth factors of the beta-class (TGFs-beta) stimulate extracellular matrix synthesis and have been implicated in embryogenesis, wound healing, and fibroproliferative responses to tissue injury. Because cells communicate with several extracellular matrix components via specific cell membrane receptors, we hypothesized that TGFs-beta may also regulate the expression of such receptors. We confirmed that TGF-beta 1 increases the expression of fibronectin, an adhesive glycoprotein expressed during embryogenesis and tissue remodeling. Based upon the 48-72-h period required for a maximal fibroproliferative response to dermal injections of TGF-beta 1, we exposed human fetal lung fibroblasts (IMR-90) to TGF beta 1 for periods up to 48 h in vitro. We observed as much as 6-fold increases in fibronectin synthesis by 24 h as previously reported for fibroblastic cells (Ignotz, R. A., and Massague, J. (1986) J. Biol. Chem. 261, 4337-4345; Ignotz, R. A., Endo, T., and Massague, J. (1987) J. Biol. Chem. 262, 6443-6446; Raghow, R., Postlethwaithe, A. E., Keski-Oja, J., Moses, H. L., and Kang, A. H. (1987) J. Clin. Invest. 79, 1285-1288), but up to 30-fold increases by 48 h. These increases are accompanied by similar increases in fibronectin mRNA levels which are prevented by actinomycin D treatment. Using a monospecific antibody raised to the human placental fibronectin receptor complex, we found that TGF-beta 1 stimulated fibronectin receptor synthesis up to 20-40-fold and increases mRNA levels encoding both the alpha- and beta-subunits up to 3-fold, compared to control IMR-90 in serum-free medium. Actinomycin D blocks TGF-beta 1-mediated increases in receptor mRNA levels. The earliest detectable TGF-beta 1-mediated increases in fibronectin receptor complex protein synthesis and mRNA levels occur at 8 h, whereas the earliest increases in fibronectin protein synthesis and mRNA levels occur at 12 h. These results demonstrate that TGF-beta 1 stimulates fibronectin receptor synthesis, extending the diverse stimulatory activities of this polypeptide to matrix receptors. In addition, because fibronectin matrix assembly may involve the fibronectin cell adhesive receptor complex, increased receptor expression may help drive fibronectin deposition into matrix. PMID- 2965147 TI - Proteolytic processing of pro-alpha and pro-beta precursors from human beta hexosaminidase. Generation of the mature alpha and beta a beta b subunits. AB - There are two major isozymes of human lysosomal beta-hexosaminidase (beta-N acetylhexosaminidase, EC 3.2.1.52), hexosaminidase A, alpha(beta a beta b), and hexosaminidase B, 2(beta a beta b). The alpha subunit contains a single polypeptide chain, while the beta subunit is composed of two nonidentical chains (beta a and beta b) derived from a common pro-beta precursor. The mature subunits, like those of most lysosomal enzymes, are produced through the proteolytic processing of propolypeptides once they enter the lysosome. In order to define the structure of the alpha and beta subunits generated in the lysosome, the alpha, beta a, and beta b polypeptides of hexosaminidase A and B were separated by a combination of molecular sieve and ion exchange high performance liquid chromatography, and amino-terminal sequences were determined. These were localized to the deduced amino acid sequences of previously isolated cDNAs coding for the prepro-alpha and beta polypeptides. From this analysis, the sites of hydrolysis generating the mature alpha, beta a, and beta b chains from hexosaminidase A and B could be determined. First, the signal peptide, required for processing of the pre-propolypeptides through the rough endoplasmic reticulum was predicted from the first in-frame Met residue on the cDNA. Second, amino acid sequencing defined the amino termini of the mature polypeptide chains and identified the pro-sequences removed from both the pro-alpha and pro-beta polypeptides. Third, an internal cleavage resulted in the removal of a tetrapeptide, Arg-Gln-Asn-Lys, and tripeptide, Arg-Gln-Asn, from the pro-beta chain of hexosaminidase A and B, respectively , to generate the beta b and beta a chains. This result localized the beta b and beta a chains to the amino-terminal and carboxyl-terminal halves of the pro-beta sequence, respectively. Finally, we previously reported minimal or no carboxyl-terminal processing of the pro-beta chain in the lysosome. On the other hand, we suggest that there is trimming at the carboxyl terminus of the pro-alpha chain based on comparison of molecular weights of deglycosylated alpha with the isolated beta b and beta a chains comprising the mature beta subunit with those predicted from the cDNA. Thus, in the lysosome the pro forms of hexosaminidase A and B undergo extensive proteolytic processing which, while specific in nature, has the appearance of removing easily accessible, nonessential domains, rather than contributing to biosynthetic maturation of function. PMID- 2965148 TI - Estradiol-stimulated turnover of heparan sulfate proteoglycan in mouse uterine epithelium. AB - Heparan sulfate proteoglycans (HSPGs) and dermatan sulfate/chondroitin sulfate proteoglycans may be extracted from the uterine epithelium of immature mice by a 1-min exposure of the luminal surface of excised uteri to 1% Nonidet P-40 detergent. In mice that are treated with estradiol there is a marked increase in free heparan sulfate glycosaminoglycan in the extract. (a) By Sepharose exclusion chromatography the [35S]sulfate-labeled major HSPG had a nominal Mr of 200-250 X 10(3), consisting of a core protein of about 80-90 X 10(3) Mr with about 8-10 heparan sulfate glycosaminoglycan chains (Mr = 13 X 10(3)). The HSPG had a lower bouyant density (less than 1.45 g/ml) than the dermatan sulfate/chondroitin sulfate proteoglycan and was heterogeneous, as was evident in the fact that HSPG attained equilibrium over a wide range of CsCl densities and also showed nonuniform interaction with octyl-Sepharose. (b) Virtually all of the major HSPG was removed when the epithelium was isolated by proteolysis, indicating a cell surface localization. A smaller, less prominent HSPG (nominal Mr = 80 X 10(3)) was synthesized during the first 2 h after isolation. (c) Label and chase experiments with and without chloroquine showed that virtually all of the free heparan sulfate glycosaminoglycan chains derived from endocytosis and lysosomal degradation of the plasma membrane-associated HSPG. We conclude that estradiol stimulates endocytosis of HSPG, predominantly from the basolateral epithelial surface and suggest that this HSPG turnover may reflect changes associated with blastocyst attachment and invasion of the endometrium. PMID- 2965149 TI - Structure of the rabbit fast-twitch skeletal muscle Ca2+-ATPase gene. AB - We have isolated two genomic clones which together encode the Ca2+-ATPase of rabbit fast-twitch skeletal muscle sarcoplasmic reticulum. One of the two 16.5 kilobase (kb) genomic inserts in the lambda phage vector Charon 4A contains 23 exons extending from the polyadenylation site at the 3' end of the ATPase gene to within 38 nucleotides of the translation initiation codon in the 5' exon. An overlapping genomic insert of 16.5 kb contains the remainder of the 5' exon and a further 8 kb of upstream sequence. S1 nuclease mapping and primer extension analysis of the 5' end of the Ca2+-ATPase mRNA indicate that the transcription initiation site is located 185 base pairs (bp) upstream of the translation initiation codon. A "TATA" box (CA-TAAA) was found at position -30 and the sequence CCAAT was found at position -78 relative to the transcription initiation site. In a previous study (Brandl, C. J., de Leon, S., Martin, D. R., and MacLennan, D. H. (1987) J. Biol. Chem. 262, 3768-3774) cDNAs for neonatal and adult forms of the fast-twitch Ca2+-ATPase were shown to encode different carboxyl-terminal sequences, presumably as a result of alternative splicing. We have now found that these different DNA sequences encoding different carboxyl terminal sequences are located in different exons. Exon boundaries of the Ca2+ ATPase gene did not correlate well with proposed domain boundaries for the Ca2+ ATPase protein. The locations of exon/intron boundaries were only partially conserved between the Ca2+-ATPase gene and a Na+/K+-ATPase gene (Ovchinnikov, Y. A., Monastyrskaya, G. S., Broude, N. E., Allikmets, R. L., Ushkaryov, Y. A., Melkov, A. M., Smirnov, Y. V., Malyshev, I. V., Dulubova, I. E., Petrukhin, K. E., Gryshin, A. V., Sverdlov, V. E., Kiyatkin, N. I., Kostina, M. B., Modyanov, N. N., and Sverdlov, E. D. (1987) FEBS Lett. 213, 73-80) and they did not follow closely the boundaries of amino acid sequences that are highly conserved among a group of ion transport ATPases. PMID- 2965150 TI - Promoter-promoter interactions influencing transcription of the yeast mitochondrial gene, Oli 1, coding for ATPase subunit 9. Cis and trans effects. AB - The gene for ATPase subunit 9 of yeast mitochondria (Oli 1) contains two promoter sequences (Op1 and Op2) separated by 78 nucleotides. Both promoters are transcribed in vivo and in vitro though with different efficiency. The upstream promoter (Op1) is 12-15 times stronger than the downstream promoter (Op2), and this difference in promoter activity is partly attributable to the influence of the +2 nucleotide (Biswas, T. K., and Getz, G. S. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 270-274). In addition, the presence of the strong promoter (Op1) in close proximity to the weak promoter (Op2) partially inhibits the expression of the latter (Op2). The relative orientation of the two promoters has no influence on these inhibitory effects. When both promoters are present in the same reaction mixture, the strong promoter always competes effectively with the weak promoter for limited RNA polymerase (trans or competition effect). When the two promoters are present in the same plasmid, there is an inhibitory interaction between them that decreases as the distance between the two promoters increases (cis or position effect). Thus, the difference between the activities of a strong and a weak mitochondrial promoter in tandem is a function of two effects, the trans or competition effect and the cis or position-related effect. A model for promoter promoter interactions is proposed. PMID- 2965151 TI - Intracellular degradation of the complement C3b/C4b receptor in the absence of ligand. AB - Human neutrophils (PMN) respond to various soluble stimuli by translocating intracellular complement C3b/C4b receptors (CR1) to the cell surface. Ligand independent internalization of surface CR1 has been demonstrated previously, but the fate of total cellular CR1 during PMN stimulation has not been determined. In order to study the fate of CR1 during neutrophil activation, we have employed a unique approach for the quantitative analysis of intracellular antigens which allows simultaneous measurement of total cellular and surface membrane antigen pools. Stimulation of isolated PMN with N-formyl-Met-Leu-Phe or ionomycin resulted in a mean 7-fold increase in surface CR1 expression within 15 min. Total cellular CR1 decreased by as much as 45% within 15 min, with loss continuing for up to 1 h. Inclusion of NH4Cl during PMN stimulation inhibited the loss of total CR1 without affecting surface CR1 expression. Addition of phenylmethylsulfonyl fluoride inhibited loss of total CR1 and enhanced the stimulus-induced increases in surface CR1. These data suggest that intracellular degradation of CR1 occurs during stimulation of PMN and may involve proteolysis in an acidic intracellular compartment. Since our experiments were done with isolated PMN in the absence of serum and complement components, this degradation occurred in the absence of C3b, the ligand for CR1. To our knowledge, ligand-independent degradation of a cell surface receptor has not been previously detected. PMID- 2965152 TI - Formation of aggregates from a thermolabile in vivo folding intermediate in P22 tailspike maturation. A model for inclusion body formation. AB - The in vivo accumulation of polypeptide chains in the form of aggregated non native states is a problem in many applications of biotechnology. In the maturation pathway of the thermostable P22 tailspike endorhamnosidase, the folding and chain association intermediates can be distinguished from the native tailspikes in crude extracts of phage-infected Salmonella cells. Temperature sensitive folding mutations, at many sites in the chain, destabilize these conformational intermediates preventing the formation of native tailspikes at restrictive temperatures (Goldenberg, D. P., Smith, D. H., and King, J. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 7060-7064). We report here that both wild type and mutant tailspike polypeptide chains which fail to reach the native state accumulate in an aggregated state. These off-pathway aggregates form from a thermolabile intermediate in the productive folding pathway. These aggregation reactions are suppressed by lowering the temperature of maturation. Similar off pathway steps from folding intermediates may account for the non-native aggregates often found in the expression of cloned genes in heterologous hosts. PMID- 2965153 TI - Inhaled ethylene oxide induces preneoplastic foci in rat liver. AB - The metabolite of E, EO, has been shown to be an extrahepatic carcinogen in rats in long-term studies. By means of a rat liver foci bioassay with 3 to 4 days old Sprague-Dawley rats, EO showed an initiating capacity in the livers of female, but not of male rats, measured as incidence of foci deficient in ATPase. After inhalation of 55 and 100 ppm EO, 8 h daily, 5 days weekly, and over 3 weeks, 1 week of pause, and another 8 weeks of promotion with polychlorinated biphenyls, foci incidence was generally low. But it was concentration dependently higher than in controls 12 weeks after starting the experiment. A linear concentration effect relationship existed with a correlation coefficient of r = 0.991. With 33 ppm EO the number of foci was not enhanced significantly. The administration of 10,000 ppm E did not result in an enhanced foci incidence. In general the carcinogenic potential of EO, which has not been shown so far to cause hepatic tumors in rats, could be demonstrated in rat liver using a sensitive rat liver foci bioassay. PMID- 2965155 TI - Plasma concentrations of medroxyprogesterone acetate and megesterol acetate during long-term follow-up in patients treated for metastatic breast cancer. AB - A total of 32 patients with metastatic breast cancer responding with at least disease stabilization to treatment with two commercially available preparations of medroxyprogesterone acetate (MPA) or one preparation of megestrol acetate (MA) were followed for their plasma concentrations. The MPA and MA were measured by HPLC. MPA from Upjohn and Farmitalia was given to 12 patients (median age, 61 years; median follow-up, 20 weeks) and 8 patients (54 years, 16 weeks), respectively, on a schedule of 1000 mg daily i.m. for 10 days followed by 200 mg t.i.d.p.o. for the remainder of the treatment course. The peak concentrations (means, 163 vs 97 ng/ml), the time to peak levels (medians, 3 vs 10 weeks), and the areas under the concentration curves from time 0 to 24 weeks (means, 2400 vs 1868 ng/ml X weeks) were significantly different in the respective treatment groups (t-test; significance level, 0.05). MA from Bristol-Myers was administered orally in one daily dose of 160 mg throughout the treatment course in 12 patients (median age, 51 years; median follow-up, 20 weeks). A mean MA peak concentration of 218 ng/ml was reached after a median of 7 days. Plateau plasma levels were higher for MA than MPA. PMID- 2965154 TI - Treatment of advanced renal cell cancer with recombinant interferon alpha as a single agent and in combination with medroxyprogesterone acetate. A randomized multicenter trial. AB - The response rates in metastatic renal cell cancer (RCC) after chemotherapy, hormonal treatment, or immunotherapy rarely exceed 15%. Recently, interferon alpha (IFN alpha) was used for treatment of this disease in several studies which also demonstrated response rates of 15%. In order to test whether IFN therapy combined with hormones would result in higher response rates we compared single agent IFN therapy with a combined therapy of rIFN alpha 2C plus medroxyprogesterone acetate (MPA) in a randomized multicenter trial. The rIFN alpha 2C (2MU) was given s.c. 5 times per week for 8-12 weeks and subsequently once weekly until week 48. In the combined treatment, 750 mg MPA was given p.o. daily until week 48 in addition to the IFN as described. The overall response rate in 93 evaluable patients was 5.4% corresponding to 2 complete and 3 partial responses. Median survival was 7 months in both treatment groups. These data confirm the ineffectivity of low IFN doses for treatment of RCC. The low response rate is not increased by addition of MPA to IFN. The analysis of other IFN studies suggests that not only IFN doses but also IFN sources may influence response rates in metastatic RCC. PMID- 2965157 TI - Potassium inhibition of transforming protein P85gag-mos and reversal of the transformed phenotype in 6m2 cells. AB - K+ at high concentrations (52-72 mM hypertonic KCl) has been reported to induce reverse transformation in the 6m2 cell, which is a clone of normal rat kidney cells (NRK) infected with a temperature-sensitive transformation virus. When exposed to high K+, 6m2 cells grown at the permissive temperature (33 degrees C) exhibit normal morphology and reduced soft agar growth, characteristics of cells grown at nonpermissive temperature (39 degrees C). In the current study, flattening of cells and rearrangement of surface microvilli were demonstrated by scanning electron microscopy to occur within 6 hr of exposure to high K+, similar to the effect of temperature shift to 39 degrees C. Exposure to K+ resulted in a 90% inhibition of P85gag-mos-associated serine kinase activity within 5 min, with a subsequent reduction of up to 75% of the synthesis of this protein. These alterations in the putative transforming protein were similar to those induced by temperature shift and were considered to be the basis for retrotransformation. The cell microtubular system and F-actin cables were affected more slowly by K+ than by a temperature shift to 39 degrees C. The former did not achieve the fine reticulum network seen in NRK cells until 72 hr later, but the latter remained aberrant. The effect on the enzyme might be mediated by alteration in phosphorylation, but the mechanism by which kinase inactivation induces retrotransformation is not yet known. PMID- 2965156 TI - Glycoprotein modifications of sarcoma L-1 tumor cells by tunicamycin, swainsonine, bromoconduritol or 1-desoxynojirimycin treatment inhibits their metastatic lung colonization in Balb/c-mice. AB - Synthesis and expression of cell surface carbohydrates appear to be involved in recognition events associated with tumor invasion and metastasis. Thus, the potential of murine sarcoma L-1 cells to form experimental lung metastases after i.v. injection was assessed after inhibiting tumor cell protein glycosylation with tunicamycin, swainsonine, bromoconduritol, or 1-desoxynojirimycin. Incubation of sarcoma L-1 cells with 0.5 microgram (or above) of these substances/ml medium for 20-24 h significantly inhibited lung colonization. Cytotoxic side effects or additional organ manifestations could not be found. Gas liquid chromatographic examinations of carbohydrates from treated L-1 cells indicated that sugar synthesis was evidently inhibited. These results suggest that specific glycan structures on tumor cells are required for expression of the metastatic phenotype. PMID- 2965158 TI - Medroxyprogesterone acetate- and ethinylestradiol-induced changes in biliary bile acids of the rat studied by high-performance liquid chromatography. AB - The effects of subcutaneous administration (5 mg/kg per day) for seven days of medroxyprogesterone acetate (MPA) or 17 alpha-ethinylestradiol (EE) on bile flow, total bile acid output and individual biliary acids have been studied in adult male Wistar rats. Biliary bile acid composition was quantitated by a simple isocratic high-performance liquid chromatographic technique using a C18 reversed phase radial compression column and refractive index detection. This method revealed that muricholic acids, analysed as taurine and glycine conjugates, constituted a higher proportion of biliary bile acids in the rat than previously observed with gas chromatographic techniques. Marked cholestasis was produced by EE while MPA had little effect on bile flow or total bile acid output. Despite this, both steroids significantly increased the proportion of taurine-conjugated muricholic acids relative to taurocholic acid, although the estrogen had the more pronounced effect. Further study of the hepatobiliary consequences of high doses of MPA would seem warranted in view of the important use of this progestogen for cancer therapy. PMID- 2965159 TI - Low dose infusions of 26- and 28-amino acid human atrial natriuretic peptides in normal man. AB - To investigate the effects of a small rise in the plasma atrial natriuretic peptide (ANP) concentration, 6 normal subjects received 2-h low dose (2 pmol/kg.min) infusions of both 28 [human alpha hANP-(99-126)]- and 26 [human ANP (101-126)]-amino acid peptides in a placebo-controlled study. Both peptides induced more than 2-fold increases in urinary sodium, calcium, and magnesium excretion. Effective renal plasma flow was slightly reduced, glomerular filtration rate did not change, and renal filtration fraction increased during the ANP infusions. Plasma renin, angiotensin II, and aldosterone concentrations fell by about 50%. Arterial blood pressure and plasma catecholamines did not change. Hematocrit and serum albumin concentrations rose significantly. ANP effects on urinary electrolytes and the renin-angiotensin-aldosterone system were sustained for over an hour after completion of the ANP infusions. The two peptides did not differ in their effects. These results are consistent with a physiological role for plasma ANP in the regulation of extracellular fluid volume and demonstrate that minor N-terminal truncation of alpha hANP has little effect on its biological activity. PMID- 2965160 TI - Identification and plasma concentrations of the N-terminal fragment of proatrial natriuretic factor in man. AB - A specific RIA was developed to measure plasma atrial natriuretic factor (ANF) N terminal immunoreactivity in man. Antibodies raised in rabbits against a rat ANF N-terminal fragment [ANF-(11-37)] had 100% cross-reactivity with human ANF-(1-30) and purified plasma N-terminal ANF immunoreactivity. The ED80 and ED50 of standard curves prepared using [125I]human ANF-(1-30) and human ANF-(1-30) were 31.5 +/- 5.4 (+/- SD) and 132.5 +/- 20.4 fmol/tube, respectively. The plasma ANF N-terminal peptide concentrations were assayed directly, without extraction, since dilution of plasma and addition of standard to plasma yielded parallel dose responses in the RIA and virtually 100% recovery of ANF-(1-30) added to plasma. Purification of ANF N-terminal immunoreactivity from 1.5 L human plasma by affinity chromatography and amino acid sequencing suggested that it was closely related to ANF-(1-98), although some degraded peptides were also detected. The mean basal plasma ANF N-terminal peptide level measured in 34 normal subjects was 420 +/- 157 (+/- SD) pmol/L. The values were higher in plasma from patients with congestive heart failure (grades III and IV; 7,041 +/- 6,136 pmol/L; n = 13) or chronic renal failure (10,079 +/- 4,942 pmol/L; n = 20). In 9 patients with chronic renal failure, hemodialysis resulted in a 30% (P less than 0.05) decrease in plasma ANF-(99-126) levels, from 34.7 +/- 12.3 (+/- SD) to 23.2 + 6.1 pmol/L, but no changes in plasma ANF N-terminal peptide concentrations. These data indicate that the N-terminal portion of pro-ANF is cosecreted with ANF-(99-126). Its higher plasma levels in the basal state and during chronic renal failure suggest a different process of elimination than that of ANF-(99-126), which may be partly mediated by the kidney. PMID- 2965161 TI - Analysis of the ATPase mechanism of myosin subfragment 1 from insect fibrillar flight muscle in the presence and absence of actin, using phosphate-water oxygen exchange measurements. AB - Phosphate-water oxygen exchange was measured during ATPase activity of myosin subfragment 1, isolated from Lethocerus flight muscle. The result supports a mechanism including a rapid reversible ATP cleavage step followed by slow Pi release, so that extensive exchange occurs by multiple reversals of the cleavage step. Interaction with actin accelerates Pi release and reduces the extent of exchange. These properties are similar to vertebrate skeletal muscle subfragment 1. Differences in kinetic properties between insect flight and rabbit skeletal muscle exhibited in the fibres, particularly in respect of the strain activation of insect flight muscle, are not exhibited in the isolated myosin heads and are therefore probably due to organization within the fibre lattice. PMID- 2965162 TI - Is atriopeptin a physiological or pathophysiological substance? Studies in the autoimmune rat. AB - Atriopeptin (AP), a natriuretic-diuretic and vasodilatory peptide, is synthesized and secreted from mammalian atria. The definitive role of this peptide on cardiovascular physiology and pathophysiology has yet to be determined. We developed a population of autoimmune rats sensitized against their own AP to evaluate the consequences of prolonged AP deficiency on physiological and pathophysiological processes. Natriuresis in response to acute intravenous volume expansion was inhibited in the autoimmune rat, however, natriuresis produced by chronic oral salt loading was not suppressed in these animals. Plasma AP increased threefold in the spontaneously hypertensive rat when evaluated as a function of blood pressure. Immunization of these rats had no effect on the rate of development, magnitude of their developing hypertension, or their daily sodium excretion when compared with nonimmunized controls. Mineralocorticoid escape occurred during desoxycorticosterone acetate administration to rats. The ability of rats to escape from the sodium-retaining effects of this steroid was not affected by prior immunization against AP. These results suggest that AP is an important natriuretic substance in response to acute intravascular volume loading. However, atriopeptin does not appear to be involved in the natriuretic response to chronic intravascular volume loading, blood pressure regulation, or mineralocorticoid escape. PMID- 2965163 TI - Ouabain-insensitive K-adenosine triphosphatase in distal nephron segments of the rabbit. AB - An electrogenic H-ATpase sensitive to inhibition by N-ethyl-maleimide has been reported to be present in renal distal tubules. In contrast to another H-ATPase (gastric H-K-ATPase), the renal enzyme is not stimulated by K+ and is not inhibited by vanadate. However, our preliminary observations indicated that a K stimulated ATPase (K-ATPase) sensitive to inhibition by vanadate is present in renal medullary collecting duct (MCD). To localize and further characterize this renal tubular K-ATPase, we measured K-ATPase activity in eight specific segments of the rabbit nephron. K-ATPase activity was the difference in ATPase activity in the presence and absence of KCl but in the presence of ouabain (to inhibit Na-K ATPase). ATPase activity was determined by a fluorometric microassay in which ATP hydrolysis is coupled to the oxidation of NADH. There was a significant K-ATPase activity (expressed as pmol.min-1.mm-1) in the connecting tubule (CNT, 17.0 +/- 3.3), cortical collecting duct (CCD, 6.6 +/- 0.7), and MCD (8.8 +/- 1.7), but not in the proximal segments and the thick ascending limbs. The renal tubular K ATPase was not only inhibited by vanadate but also by omeprazole and SCH 28080 (relatively specific inhibitors of gastric H-K-ATPase). It is concluded that K ATPase present in the CNT, CCD, and MCD has some properties in common with gastric H-K-ATPase. However, the physiological role of K-ATPase in the distal nephron segments remains to be elucidated. PMID- 2965164 TI - Lymphocytes expressing type 3 complement receptors proliferate in response to interleukin 2 and are the precursors of lymphokine-activated killer cells. AB - In the absence of antigenic or mitogenic stimulation, certain peripheral blood lymphocytes exhibit proliferative and lymphokine-activated killer (LAK) cell activities when cultured with recombinant IL-2. Both activities were found to be an exclusive property of lymphocytes expressing type 3 complement receptors (CR3) identified by anti-CD11 monoclonal antibodies. CD11+ lymphocytes were then fractionated into three subsets by two-color flow cytometry. These included CD16+ cells, which display distinctive Fc receptors for IgG (CD16). Using anti-CD5, the CD11+ CD16- lymphocytes were separated into non-T cell and T cell subsets. The two non-T cell subsets (CD11+ CD16+ and CD11+ CD16- CD5-), but not the T cell subset (CD11+ CD16- CD5+), could proliferate in response to IL-2. Both CD11+ non T cell subsets, but not the CD11+ T cell subset, had the capacity to mediate natural killer cell activity. However, all three CD11+ lymphocyte subsets were capable of generating LAK activity. These findings are consistent with the concept that two signals are required to stimulate T cells to proliferate. However, at least a small subset of blood T cells can be activated by IL-2 to become LAK cells. PMID- 2965165 TI - The pharmacokinetics of oral isradipine in normal volunteers. AB - The pharmacokinetics and relative bioavailability of oral isradipine, a dihydropyridine calcium channel blocking agent, were determined in 42 normal male volunteers participating in two separate studies. Eighteen of the subjects received 2.5-, 5-, and 10-mg oral doses of isradipine solution (Study 1). The remaining 24 subjects received four 2.5-mg capsules, one 10-mg capsule, and 10 mg of isradipine as an oral solution (Study 2). Venous blood samples were obtained prior to and at frequent intervals after administration of each dose form. Plasma isradipine concentrations were measured by radioimmunoassay. No significant dose effect occurred with respect to any pharmacokinetic parameter except AUC and Cmax in Study 1. In Study 2, Cmax, tmax, and MRT were significantly different after the solution compared with the capsular formulations. The respective pharmacokinetic parameters (mean +/- SD) for the 10-mg solution and 10-mg capsule in Study 2 were time to maximum concentration, 0.40 +/- .28 and 1.57 +/- 0.44 hours; oral clearance, 284.9 +/- 105.3 and 317.0 +/- 138.4 L/hr; elimination half life, 5.36 +/- 1.8 and 6.63 +/- 2.4 hrs, respectively. Headache, dizziness, and tachycardia were the most frequent adverse effects in both studies. PMID- 2965166 TI - Nonatopic eczemas. PMID- 2965167 TI - Roaccutane and menorrhagia. PMID- 2965168 TI - Treatment of Papillon-Lefevre syndrome with etretinate. PMID- 2965170 TI - Access: meeting the needs of special patients. PMID- 2965169 TI - Analysis of orthodontic treatment by pediatric dentists and general practitioners in Indiana. AB - This investigation analyzed the extent of orthodontic services currently being provided by pediatric dentists and general practitioners in Indiana. All 71 Indiana pediatric dentists primarily in private practice and 500 general practitioners were mailed a survey/questionnaire. Sixty-two percent of the pediatric dentists and 17.9 percent of the general practitioners surveyed provided comprehensive orthodontic treatment, results much higher than those of previous surveys of pediatric dentists and general practitioners. Pediatric dentists were found to provide significantly more comprehensive orthodontic treatment and spend significantly more time providing orthodontic treatment than general practitioners, especially those in communities of less than 25,000. Sixty three percent of the practitioners surveyed had taken some type of continuing education in orthodontics. These practitioners spent significantly (p less than 0.001) more time treating orthodontic conditions, and treated more complex cases than those who had not taken such courses. PMID- 2965171 TI - Late restenosis after emergent coronary angioplasty for acute myocardial infarction: comparison with elective coronary angioplasty. AB - The late restenosis rate after emergent percutaneous transluminal coronary angioplasty for acute myocardial infarction was assessed by performing outpatient follow-up cardiac catheterization in 79 (87%) of 91 consecutive patients who had been discharged from the hospital with a successful coronary angioplasty. The majority of patients (90%) received high dose intravenous thrombolytic therapy with streptokinase in addition to angioplasty. Similar follow-up data were obtained in 206 (90%) of 228 consecutive patients who had successful elective angioplasty during the same period. The interval from angioplasty to follow-up was 28 +/- 9 weeks for the myocardial infarction group and 30 +/- 11 weeks for the elective group. Baseline clinical variables were similar for both the myocardial infarction and elective groups except for a higher percentage of men in the infarction group (81 versus 63%, p = 0.001). The number of coronary lesions undergoing angioplasty and the incidence of intimal dissection were similar, but multivessel angioplasty was more common in the elective group (13 versus 4%, p = 0.02). The rate of in-hospital reocclusion was higher in the patients receiving angioplasty for myocardial infarction (13 versus 2%, p = 0.0001). At the time of late follow-up after hospital discharge, the patients with myocardial infarction were more often asymptomatic (79 versus 55%, p = 0.0001), and the rate of angiographic coronary restenosis was lower for the infarction group both overall (19 versus 35%, p = 0.006) and when multivessel angioplasty patients were excluded (19 versus 33%, p = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965172 TI - Repeat coronary angioplasty: correlates of a second restenosis. AB - To identify the correlates of a second restenosis after repeat percutaneous coronary angioplasty, the records of 384 patients with single vessel disease who underwent repeat angioplasty for restenosis complicating a first elective angioplasty were examined. A second restenosis occurred in 47 (31%) of 151 patients having angiographic follow-up. Univariate correlates of a second restenosis were an interval between the first and second angioplasty less than 5 months (41 versus 20% of patients had restenosis, p less than 0.01), male gender (35 versus 12%, p less than 0.05), lesions length greater than or equal to 15 mm before the second angioplasty (62 versus 28%, p less than 0.05), diameter stenosis greater than 90% before the second angioplasty (67 versus 29%, p less than 0.05), final gradient greater than 20 mm Hg after the second angioplasty (52 versus 28%, p less than 0.05) and an additional site requiring dilation at the time of the second angioplasty (50 versus 29%, p = 0.10). Multivariate predictors of a second restenosis were an interval of less than 5 months between the first and the second angioplasty (p = 0.001), male gender (p = 0.001), lesion length greater than or equal to 15 mm before the second angioplasty (p = 0.001) and the need to have an additional site dilated at the time of the second angioplasty (p = 0.002). Patients at increased risk of restenosis after the second angioplasty can be identified and may serve as a useful population for intervention studies. PMID- 2965173 TI - Correlation between beta-endorphin plasma levels and anginal symptoms in patients with coronary artery disease. AB - To verify whether beta-endorphin plasma levels influence the presence of anginal symptoms, 74 consecutive male patients were studied. All patients had previously documented coronary artery disease and reproducible exercise-induced myocardial ischemia. Thirty-five patients (Group I) had a history of angina and reported anginal symptoms during exercise stress testing; 39 patients (Group II) were asymptomatic and had documented silent myocardial ischemia during exercise. Baseline beta-endorphin plasma levels were measured in blood samples taken before exercise stress testing and analyzed by beta-endorphin-I125-RIA Kit-NEN (a radioimmunoassay method). The mean baseline beta-endorphin plasma level was 22.5 +/- 19 pg/ml in patients with anginal symptoms compared with 43.7 +/- 28 pg/ml in asymptomatic patients (p less than 0.001). Baseline blood pressure and heart rate systolic pressure (rate-pressure) product at baseline and at ischemia threshold (1 mm ST segment depression) were similar in the two groups. Group II patients had a longer exercise duration (p less than 0.01), more pronounced ST segment depression (p less than 0.001) and a higher peak rate-pressure product (p less than 0.01). The extent of coronary artery disease, ejection fraction and left ventricular end-diastolic pressure were similar in the two groups. These data suggest that higher baseline beta-endorphin plasma levels may play a role in the decreased sensitivity to pain in patients with silent myocardial ischemia. In addition, different beta-endorphin levels can be associated with a different sensitivity to pain. PMID- 2965174 TI - Assessment of myocardial perfusion in humans by contrast echocardiography. I. Evaluation of regional coronary reserve by peak contrast intensity. AB - Myocardial contrast echocardiography was performed during coronary angiography with 2 ml of sonicated meglumine diatrizoate sodium 76% (meglumine) in 40 patients (ranging in age from 25 to 79 years) before and 10 to 15 s after intracoronary injection of papaverine, 8 mg into the right coronary artery (n = 43) and 10 mg into the left (n = 46). The same protocol was repeated in 17 patients 5 to 10 min after completion of coronary angioplasty. In 13 patients with normal coronary angiograms, peak contrast intensity corrected for background myocardial intensity was measured in 36 regions and was found to increase after papaverine from 36 +/- 16 to 55 +/- 22 U (p less than 0.001). In contrast, in the 27 patients with angiographic evidence of coronary artery disease, peak intensity in 64 regions remained unchanged after papaverine (35 +/- 22 versus 36 +/- 23 U). An increase in peak intensity greater than or equal to 10 U was 80% sensitive and 92% specific for coronary artery disease. After successful coronary angioplasty, peak intensity in the involved regions improved significantly (p less than 0.001) during baseline contrast injections (from 32 +/- 16 to 50 +/- 25 U) as well as in the postpapaverine contrast injections (from 30 +/- 12 to 60 +/- 26 U). In conclusion, measurement of peak contrast intensity after intracoronary injections of sonicated meglumine provides a relative index of myocardial perfusion that allows assessment of regional coronary reserve in patients with coronary artery disease. This may be of particular value in evaluating the immediate effects of coronary angioplasty on myocardial perfusion. PMID- 2965175 TI - Effects of changes in coronary stenosis on left ventricular diastolic filling assessed with pulsed Doppler echocardiography. AB - To determine the effects of changes in coronary stenosis on left ventricular diastolic filling, diastolic filling was serially examined before and after percutaneous transluminal coronary angioplasty using pulsed Doppler echocardiography in 50 patients with stable exertional angina pectoris. Peak rapid filling velocity and the ratio of peak atrial filling to peak rapid filling velocities were measured from the transmitral flow velocity pattern before and 2 and 9 days after coronary angioplasty. Peak rapid filling velocity increased and the ratio of peak atrial filling to peak rapid filling velocities decreased gradually after coronary angioplasty. The improvement in left ventricular diastolic filling was greater in patients with severe (greater than 90%) coronary stenosis than in patients with mild (less than or equal to 90%) coronary stenosis. In the long-term follow-up period, the improved left ventricular diastolic filling worsened in only 11 patients with marked progression to greater than 90% coronary stenosis. Thus, left ventricular diastolic filling improved gradually after coronary angioplasty, possibly reflecting post-ischemic "stunned" myocardium. Serial examinations of left ventricular diastolic filling with pulsed Doppler echocardiography may be a means of noninvasively assessing the temporal changes in the coronary stenosis and predicting the occurrence of coronary restenosis after coronary angioplasty. PMID- 2965176 TI - Affinity purification of hexosaminidases. AB - Hexosaminidases A and B were purified by affinity chromatography from normal gastric mucosa, after preliminary separation of isozymes by anion exchange chromatography. Heparin and mannosamine were coupled to Sepharose 4B and used as affinity matrices and the purified enzymes were found to be homogeneous when analysed by polyacrylamide slab gel electrophoresis. This combination of 2 novel affinity chromatographic procedures is superior to existing methods in that a final yield of over 70% could be achieved. Also, the number of steps required to obtain homogeneity are less in contrast to the conventional methods used previously. PMID- 2965177 TI - Diuretic, natriuretic and hypotensive effects of synthetic atrial natriuretic factor in conscious newborn calves. AB - The effects of synthetic Atrial Natriuretic Factor (ANF) on urine flow rate, sodium excretion, potassium excretion and arterial blood pressure were studied in 10-12 days-old female calves. In four female calves fitted with a Foley catheter, an intravenous administration of ANF (Ile-ANF 26; 1.6 micrograms/kg body wt during 30 min) induced an increase (P less than 0.01) in urine flow rate (from 1.8 +/- 0.2 to 12.8 +/- 1.1 ml/min), sodium excretion (from 0.15 +/- 0.02 to 0.81 +/- 0.06 mmol/min) and free water clearance (from 0.13 +/- 0.9 to 5.16 +/- 0.5 ml/min). It had no significant effect on potassium excretion. In four calves chronically-instrumented with a carotid catheter, an intravenous administration of synthetic ANF alone (1.6 micrograms/kg body wt during 30 min) induced a gradual decrease (P less than 0.01) in systolic, diastolic and mean arterial blood pressure (from 112 +/- 4 to 72, from 72 +/- 2 to 61 +/- 1 and from 90 +/- 2 to 65 +/- 2 mmHg respectively, at the end of ANF infusion). An intravenous administration of angiotensin II (AII) (0.5 micrograms/kg body wt during 45 min) induced a significant increase in systolic, diastolic and mean arterial blood pressure which was antagonized by an i.v. bolus injection of ANF (0.125 micrograms/kg body wt). However, during a simultaneous administration of AII (0.3 micrograms/kg body wt during 30 min) and ANF (1.6 micrograms/kg body wt. during 30 min), the atrial peptide did not influence the pressure action of AII. These findings indicate that the conscious newborn calf is sensitive to diuretic, natriuretic and hypotensive effects of synthetic ANF. PMID- 2965178 TI - Chronic low-dose infusions of dexamethasone in rats: effects on blood pressure, body weight and plasma atrial natriuretic peptide. AB - Glucocorticoid-induced hypertension in rats has been studied using long-term, low dose dexamethasone treatment. Dose-related increases in systolic blood pressure were achieved, without loss in body weight, with subcutaneous continuous infusions of 1, 2 and 5 micrograms dexamethasone per day, respectively, for 4 weeks. Rats treated with 10 micrograms dexamethasone per day lost weight at a rate of 10 g per week. Lower doses caused a significant reduction in weight gain compared with controls. Renin, aldosterone, plasma sodium and potassium concentrations were unaffected by dexamethasone treatment. Plasma atrial natriuretic peptide (ANP) concentrations were decreased by 40-50% by dexamethasone. These decreases were negatively correlated with increases in systolic blood pressure and haematocrit. Glucocorticoid-induced decreases in ANP contrast with ANP increases in response to mineralocorticoid treatment in rats with deoxycorticosterone-induced hypertension. Plasma concentrations of the endogenous glucocorticoid, corticosterone, were suppressed to the same very low levels by 5 and 10 micrograms dexamethasone per day; 1 and 2 micrograms doses were less effective. Unlike mineralocorticoid-induced hypertension, the pressor effects of dexamethasone were ameliorated but not abolished by dietary sodium restriction and were unaffected by sodium loading. Two micrograms of dexamethasone reduced plasma ANP in rats on either high- or low-sodium diets by 29 and 34%, respectively. We conclude that low-dose infusions (less than 5 micrograms/day) of dexamethasone are suitable for studying glucocorticoid-induced hypertension without the complications of weight loss that have been reported by others or of the mineralocorticoid-like side effects which endogenous glucocorticoids may exhibit. PMID- 2965179 TI - The expression of B cell surface receptors. I. The ontogeny and distribution of the murine B cell IgE Fc receptor. AB - The ontogenic appearance and lymphoid tissue distribution of the murine B cell IgE FcR (Fc epsilon R) was examined. Flow cytometry was utilized to study the expression of the Fc epsilon R on splenic B cells from mice of increasing age, as well as B cells from various lymphoid organs. A large panel of B cell tumors was also screened for the presence of the Fc epsilon R. The results demonstrate that the Fc epsilon R appears very late in B cell development, and is preceded in appearance even by IgD. In adult animals, the Fc epsilon R was found to be expressed on virtually all mature IgM, IgD bearing B cells, whether taken from the spleen, lymph nodes, or Peyer's patches. Further examination showed that B cells which had switched to express an isotype other than IgD, appeared to no longer display the Fc epsilon R. When surveying a variety of B cell tumors, the Fc epsilon R was found to be present on WEHI 279, an IgM, IgD-bearing lymphoma. The receptor was not found on pre-B cell, immature B cell, switched B cell, or secreting B cell tumors. Taken together, these results indicate that the B cell Fc epsilon R is expressed predominantly on mature, virgin B cells, and is lost after activation and switching. PMID- 2965180 TI - Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells. AB - A group of mAb recognizing the 200- and/or 220-kDa determinants (CD45R) of the leukocyte common Ag such as 2H4, WR16, MD4.3, and SN130 cross-block each other showing that they recognize a closely related epitope. The antibody UCHL1 reacts with a 180-kDa determinant of the leukocyte common Ag and exhibits a reciprocal T subset distribution pattern to the CD45R group. Peripheral blood T cells were 40% positive for UCHL1 and 58 to 65% positive for the CD45R antibodies; less than 1% of cells stained for both. On activation of CD45R+,UCHL1- T cells by PHA, up to 40% of cells became positive for both CD45R and UCHL1 by day 3. By day 7, CD45R+,UCHL1- cells fell from 90 to less than 21% whereas UCHL1+,CD45R- cells rose from 2 to 93%. Conversely, PHA-stimulated UCHL1+,CD45R- cells remained UCHL1+,CD45R- during the 7 days in culture showing that phenotypic change was unidirectional from CD45R+ to UCHL1+. In primary allogeneic mixed lymphocyte reactions, activated CD45R+ T cells also showed a change to UCHL1+. When these cells were rechallenged by the original alloantigen, the UCHL1+ cells showed 7- to 20-fold greater proliferation than the CD45R+ cells on day 3 after rechallenge. The recovery of virtually all alloantigen induced secondary proliferative response in the UCHL1+,CD45R- T cell population suggests that UCHL1 identifies a primed population of T cells which may include memory cells. PMID- 2965181 TI - Arginine vasopressin (AVP) replacement of helper cell requirement in IFN-gamma production. Evidence for a novel AVP receptor on mouse lymphocytes. AB - Arginine vasopressin (AVP), a nine-amino acid neurohypophyseal hormone, is capable of replacing the helper cell requirement for IFN-gamma production by Lyt 2+ mouse splenic lymphocytes. We present data here showing that the AVP helper signal occurs via interaction with a novel R on splenic lymphocytes and involves primarily the N-terminal six-amino acid cyclic ring (pressinoic acid) with the C terminal three-amino acid end of AVP playing a minor role. Pressinoic acid was capable of providing help at concentrations similar to those of AVP, whereas oxytocin and isoleucine pressinoic acid were 10- and 100-fold less effective, respectively. Isoleucine pressinoic acid has the same structure as pressinoic acid except for the substitution of isoleucine for phenylalanine in position 3 of the sequence. Consistent with the function data, R binding competitions with splenic lymphocyte membrane preparations showed that AVP and pressinoic acid competed similarly with [3H]AVP, whereas oxytocin and isoleucine pressinoic acid were much less effective competitors. Further characterization of the AVP lymphocyte R was performed using AVP analogues having well defined agonist and antagonist activities on either V1 (vasopressor) R or V2 (antidiuretic) R. The AVP helper signal was blocked by the V1 antagonist [d(CH2)1(5) Tyr(methyl)]AVP but not by another V1 antagonist, [d(CH2)1(5)D-Tyr(ethyl)2Val4]AVP. Both V1-R antagonists were able to block [3H]AVP binding to the V1-R on liver cells, whereas only the V1 antagonist that blocked AVP help was able to compete effectively for the spleen AVP-R. Neither a V2 agonist nor a V2 antagonist had any effect on AVP help in IFN-gamma production. These data strongly indicate the presence of a novel AVP-R on spleen lymphocytes, which is related to the classic V1-R on liver cell membranes. PMID- 2965182 TI - Glial retinal Muller cells produce IL-1 activity and have a dual effect on autoimmune T helper lymphocytes. Antigen presentation manifested after removal of suppressive activity. AB - The Muller cell is the major glial element of the mammalian retina. It can be induced to express MHC class II determinants in culture, when exposed to supernatant from activated lymphocytes. In an in vitro co-culture system, Muller cells had been shown to exert a profound inhibitory influence on Ag- and IL-2 driven proliferation of Th cell lines. In the present report we demonstrate that Muller cells can produce IL-1 activity, and that in conditions where their inhibitory action is suppressed, they display the capacity to efficiently function as APC. Mildly trypsinized and glutaraldehyde-fixed Muller cells that had been preincubated with Ag and IFN-gamma-containing rat spleen conditioned medium were able to present Ag to an S-Ag specific Th lymphocyte line, which has the capacity to induce a severe uveoretinitis in rats. Addition of the Ag after fixation did not result in presentation, showing that Muller cells had to process the nominal Ag into an immunogenic form. It was also found that the efficiency of presentation was greatly increased in the presence of naive syngeneic or allogeneic accessory cells. This enhancing effect could not be reproduced with rIL-1. This report extends our ongoing study of interactions between the retinal glial Muller cell and T lymphocytes. The results show that these interactions can be diverse, even opposite in different conditions, and suggest the possibility that Muller cells could play a determining role in the course of immune reactions at the level of the neuro-retina. PMID- 2965183 TI - The 2H4 (CD45R) antigen is selectively decreased in multiple sclerosis lesions. AB - To analyze expression of the 2H4 (CD45R) Ag on inflammatory cells in the central nervous system immune response, immunohistochemical staining with a panel of anti T cell mAb was performed on central nervous system tissues from 12 patients with multiple sclerosis (MS) and 8 patients with viral encephalitis. Only rare cells were stained with anti-2H4 in MS plaques, plaque edges, and adjacent white matter, whereas 2H4+ cells were more numerous in viral encephalitis (p less than 0.001). By contrast, no quantitative differences were found between MS plaque edges and viral encephalitis with anti-4B4 (helper-inducer function associated), anti-CD4, anti-CD3, and anti-IL-2R mAb, although there were fewer CD8+ cells in MS (p less than 0.01). These data indicate that the 2H4 Ag is selectively decreased and, because it is associated with suppressor-inducer function of CD4+ cells, that there may be a defect in the down-regulation of the in situ immune response in MS. PMID- 2965184 TI - CD4+ T cell lines with selective patterns of autoreactivity as well as CD4- CD8- T helper cell lines augment the production of idiotypes shared by pathogenic anti DNA autoantibodies in the NZB x SWR model of lupus nephritis. AB - The (NZB x SWR)F1 hybrid mice (SNF1) uniformly develop lethal glomerulonephritis in marked contrast to their parents and produce nephritogenic autoantibodies that consist of highly cationic, IgG anti-DNA antibodies that share distinct cross reactive idiotypes called IdLNF1 (idiotypes-lupus nephritis-SNF1). Herein we found that spleen cells of SNF1 mice at the late prenephritic stage, contained CD4+/CD8- and CD4-/CD8- Th that not only induced their B cells in vitro to produce highly cationic IgG autoantibodies to DNA but IdLNF1-positive IgG antibodies as well. The double-negative Th were unexpected in the SNF1 mice because they lack the lpr (lymphoproliferation) gene. We also derived IL-2 dependent CD4+/CD8- as well as CD4-/CD8- T cell lines from nephritic SNF1 mice, that could simultaneously induce IdLNF1-positive and cationic anti-DNA antibodies of IgG class. The CD4+ T cell lines consisted of "autoreactive" T cells, but not all of the lines were equal in autoantibody-inducing capability. Remarkably, the T cell lines that preferentially responded to F1-hybrid-MHC determinants, had a significantly greater ability to augment the production of pathogenic autoantibodies. The SNF1-Th could also augment autoantibody production by the NZB or SWR parent's B cells; however, IdLNF1-positive and cationic anti-DNA autoantibodies of IgG class were not induced, suggesting that the SNF1 mice possess a select population of inducible (susceptible) B cells that are committed to produce nephritogenic autoantibodies and the parental strains are deficient in such B cells. Thus, production of nephritogenic autoantibodies with IdLNF1 markers in the SNF1 mice could result from an interaction between a select population of B cells and CD4+ Th that preferentially recognize unique F1-hybrid MHC determinants, as well as double-negative auxiliary Th. PMID- 2965186 TI - L3T4+ T cells regulate Abelson virus-induced lymphomagenesis. AB - To evaluate the role of T cells in regulation of lymphomagenesis, experiments were performed using Abelson murine leukemia virus (AMuLV). In vitro transformation of bone marrow target cells by this B lymphotropic retrovirus was inhibited by peripheral lymph node cells from naive mice. The inhibitory activity depended on Thy-1+ L3T4+ cells but did not require Lyt-2+ cells. In vivo depletion of L3T4+ T cells with a mAb (GK1.5) altered the course of AMuLV-induced lymphoma. L3T4 depletion of naturally resistant C57BL/6 mice resulted in dramatic susceptibility to lymphoma induction. Lymphoma cells from anti-L3T4-treated C57BL/6 mice infected with AMuLV displayed the B lineage transformation marker P1606C3. These studies reveal an important immunologic component of Abelson disease resistance involving L3T4+ T cells. PMID- 2965185 TI - Suppression of simian immunodeficiency virus replication in vitro by CD8+ lymphocytes. AB - The AIDS-like disease in rhesus monkeys induced by the simian immunodeficiency virus (SIV) has been used as a model to explore the nature of the T lymphocyte response after infection with viruses of the human immunodeficiency virus family. Activated CD8+ lymphocytes are present in increased numbers in the paracortex of lymph nodes of SIV-infected rhesus monkeys with a lymphadenopathy syndrome. We demonstrate that SIV is more readily isolated from CD8+ lymphocyte-depleted PBL of SIV-infected animals than from their unfractionated PBL. Rather than reflecting the fact that the CD8+ lymphocyte-depleted cell populations are simply enriched for CD4+ lymphocytes, this indicates that CD8+ cells themselves are critical in this regulatory interaction. In fact, CD8+ lymphocytes from SIV infected but not uninfected rhesus monkeys can block SIV replication in vitro in PBL populations. A T lymphocyte population that blocks replication of viruses of the HIV family may contribute to containing the progression of AIDS. PMID- 2965187 TI - A C terminus cysteine of diphtheria toxin B chain involved in immunotoxin cell penetration and cytotoxicity. AB - The role of diphtheria toxin (DT) B-chain subdomains in DT cytotoxicity and immunotoxin mechanism of action has been investigated. OKT3 (mAb to the CD3 surface Ag of human T lymphocytes) was conjugated to DT or the DT mutant CRM 1001, which has a cys----tyr substitution at position 471 of the B chain. OKT3 CRM 1001 immunotoxin was about 1400-fold less cytotoxic for CD3 Jurkat cells than OKT3-DT and had a 12-fold slower kinetics of protein synthesis inactivation, CRM 1001 killed DT-sensitive Vero cells at a 5000-fold higher concentration than DT. Its cell surface-binding activity was comparable to DT. Based on kinetics of cell inactivation, toxicity determination at low extracellular pH and Triton X-114 distribution, it was concluded that CRM 1001 is defective in at least one crucial step of toxin penetration and is unable to cross cell membranes as efficiently as DT. The substituted cysteine appears to be important for DT translocating functions. Data on the function of DT B-chain subdomains are relevant for the study of whole toxin conjugates and their mechanism of action. PMID- 2965188 TI - Isolation of rat/rat T cell hybrids using W/Fu C58(NT)D as fusion partner. AB - Functional rat/rat T cell hybrids were isolated for the first time by the fusion of spleen cells from rats tolerized to the hapten TNP to a HAT-sensitive rat thymoma (C58(NT)D). 11 fusions using different protocols were attempted to assess the optimal conditions for high hybridization frequency of the desired specificity. Interestingly, the cell density requirement of the non-transformed fusion partner took precedence over that of the C58 cell line, i.e., rat cells needed to be at high density after fusion, but others have reported that mouse cells prefer a much lower density even when the same line (C58) is used. Six fusions yielded hybridomas with between 3% and 70% of wells containing hybrids after three weeks of culture, depending on the protocol used. Phenotypically, all of the hybrids and clones tested expressed the W3/25 (rat CD4) antigen, but no OX 8 (rat CD8) or immunoglobulin molecules. A minority of hybrids were found to secrete factors able to suppress (a) proliferation, (b) antibody production, and (c) cells bearing IL-2 receptors, but none appeared to suppress the production of IL-2 itself. In contrast to non-transformed rat T cell lines, the T hybrids isolated were easy to grow to high densities, clone and freeze without the need to add exogenous antigen or lymphokines to the cultures at any stage. PMID- 2965189 TI - [Beta-endorphin-like immunoreactivity in peripheral blood, especially in erythrocytes during delivery]. AB - Recently it was reported that beta-endorphin-like immunoreactivity (beta-ELI) was detectable not only in plasma, but also in erythrocytes. To assess the presence of beta-ELI in human erythrocytes and also to determine if erythrocyte-associated beta-ELI may be released into plasma in stress situations, beta-ELI concentrations in plasma and erythrocytes were measured in pairs in normal menstrual cyclic women, postmenopausal women and pregnant women during delivery. After separating from plasma, erythrocytes pellets were lysed by adding distilled water. Extraction of beta-ELI in plasma and RBC lysate was performed with a Sep pack C18 cartridge (Waters Associates Inc.), and beta-ELI concentrations were measured with a New England Nuclear radioimmunoassay kit. The following results were obtained: 1) There certainly are two pools of beta-ELI; one in plasma, the other erythrocyte-associated. 2) During delivery beta-ELI concentrations in plasma showed a great change, but beta-ELI concentrations in erythrocytes remained almost unchanged. From these results it is unlikely that erythrocytes act as a peripheral reservoir of circulating beta-ELI. PMID- 2965190 TI - The impact of AIDS on corneal transplantation. PMID- 2965191 TI - The use of heparin locks (PRN adapters) VS. intravenous therapy in outpatient surgery. PMID- 2965192 TI - Operating room nursing protocol for the patient undergoing planned local anesthesia. PMID- 2965193 TI - Atrial natriuretic peptide inhibits angiotensin II-induced contraction of isolated glomeruli and cultured glomerular mesangial cells of rats: the role of calcium. AB - We examined the contractile effect of angiotensin II on isolated rat glomeruli and cultured glomerular mesangial cells, as well as the possible involvement of calcium in this contraction. In addition, the effect of rat atrial natriuretic peptide (ANP) on angiotensin II-induced contraction was studied. Computer assisted image analysis was used to measure glomerular cross-sectional area and planar mesangial cell surface area in several experimental conditions. Angiotensin II induced a decrease in glomerular cross-sectional area and planar mesangial cell surface area that was both time and dose dependent. This effect was partially blocked by preincubation with verapamil or TMB-8, the latter compound being more effective. Incubation with ANP prevented angiotensin II induced reduction of glomerular cross-sectional area and planar mesangial cell surface area and reversed contraction in mesangial cells. Incubation with TMB-8 impaired the effect of ANP. These data suggest that angiotensin II could exert part of its physiologic effects in the kidney through calcium-dependent mesangial and glomerular contraction. ANP inhibited this contraction, possibly explaining some of its renal effects. PMID- 2965194 TI - The roles and functions of coordinators of handicapped services in higher education. PMID- 2965195 TI - Human natural killer cells enhance a mixed leukocyte reaction. AB - Natural killer cells (NK) have been reported to down-regulate the initiation of T cell responses in animal models. In the current study, highly purified CD16+ human NK cells were obtained by cell sorting and their effect on the stimulation of allogeneic T cells (MLR) determined. NK cells did not directly stimulate T cell proliferation. However, when added to a population of loosely adherent mononuclear cells (LAM), NK enhanced the ability of these accessory cells to stimulate T proliferation. This effect was not reproduced by the addition of sorted CD5 + T cells, sorted CD16- cells, or control lymphocytes to the MLR. The effect of NK on the MLR was not restricted by class II antigens and was similar to the effect of adding IL-1 to MLR cultures. These results demonstrate that human NK cells are capable of enhancing a T cell response. PMID- 2965196 TI - Enrichment of dendritic cells from human peripheral blood. AB - A simplified method is described for purification of dendritic cells from human peripheral blood. The method is based on depletion of phagocytes with carbonyl iron and magnet, followed by centrifugation of nonphagocytic cells on Percoll and elimination of contaminating T lymphocytes, B lymphocytes, natural killer cells, and monocytes from the low-density cell fraction by treatment with monoclonal antibodies and complement. The purity of enriched dendritic cells was about 80% and these cells represented 0.2% of the starting mononuclear cell population. Dendritic cells were potent autologous and allogeneic stimulators in mixed leukocyte cultures. PMID- 2965197 TI - Effect of lead on macrophage function. AB - Lead (Pb) has been shown to alter various parameters of immune function such as host resistance and antibody formation. In addition, various heavy metals have been implicated as inducers of autoimmunity. In these experiments, macrophages, isolated from the peritoneal cavity of mice exposed to various doses of lead in vivo as well as cells exposed in vitro were tested for the following immunologic parameters: phagocytosis, antigen presentation, interleukin 1 production, and their ability to stimulate the autologous mixed lymphocyte reaction (AMLR). The results obtained indicate that Pb appears to alter the ability of macrophages to present antigen by enhancing the AMLR while having no effect on phagocytosis or IL-1 production. These data suggest that Pb may interfere with antigen-specific interactions between macrophages and T cells. PMID- 2965198 TI - Pharmacological modulation of localized inflammatory reactions: the nonsteroidal anti-inflammatory drug as an adjunct to therapy. AB - An experimental model was utilized to examine the physiological effects of nonsteroidal anti-inflammatory compounds on locally induced inflammatory lesions in laboratory rabbits. The modulation of specific parameters associated with the inflammatory response, was monitored in vivo using radiolabeled cells and proteins, following the local administration of either indomethacin (INDO), acetylsalicylic acid (ASA) or sterile saline (as control) in the dermis. In one half of the dorsal aspect of each animal, Arthus-type dermal lesions were induced in triplicate, while similar sites were prepared at identical time periods (1-4, and 6 h) on the contralateral halves. In these latter sites, either INDO or ASA was administered (100 micrograms/site). The non-drug-treated lesions served as intrinsic controls when the inflammatory parameters of vascular permeability, neutrophil (polymorphonuclear) leukocyte infiltration and hemorrhage were investigated. Also, sterile saline or drug-injected, noninflamed sites prepared in each half served as further controls. Statistical analysis indicated a significant suppression of the above-named inflammatory parameters when either INDO or ASA was applied to localized inflammatory sites, as compared to non-drug treated lesions. This study may serve to illustrate the value of deploying nonsteroidal anti-inflammatory preparations, topically, when conservatively managing the acute patient in clinical practice. PMID- 2965199 TI - An analytical survey of structural aberrations observed in static radiographic examinations among acute low back cases. AB - For many years, static standing x-ray views of the lumbar spine and pelvis have been evaluated to identify variations in the positional attitude of the osseous segments and to relate such findings with possible causes of low back pain and disability. This study of 110 cases of acute low back pain identified the frequency of occurrence of intervertebral disc-space wedging, the level where it most frequently occurred and the associated alterations in the attitude of the pelvis and adjacent vertebral segments. Correlation of the location of pain with the distorted structural positions was also attempted. The study suggested significant reliability in the analysis of static x-ray views of the lumbar spine in locating the level of lesion in acute low back cases, and sought to differentiate between significant structural change and normal postural accommodations. PMID- 2965200 TI - Need for multiple diagnosis in the presence of spondylolisthesis. AB - A case of sacroiliac syndrome and lumbar facet capsulitis coexisting with an asymptomatic grade II spondylolisthesis is presented. Spinal manipulative therapy is not contraindicated in the presence of spondylolisthesis if specific to the site of joint hypomobility and if the underlying spondylolisthesis is stable. Manipulation may prove to be diagnostic as well as therapeutic. The need to clinically assess whether the spondylolisthesis is symptomatic is discussed. PMID- 2965201 TI - Back school and chiropractic practice. PMID- 2965202 TI - The efficacy between spectinomycin and norfloxacin in the treatment of uncomplicated gonorrhoeae. PMID- 2965203 TI - Aortic valve disease and the ST segment/heart rate relationship: a longitudinal study before and after aortic valve replacement. AB - The ST segment/heart rate relationship or maximal ST/HR slope has been validated as an index of myocardial ischemia in selected populations of patients with angina pectoris. The present study involved patients selected as having aortic valve disease unaccompanied by angiographic coronary artery narrowing. In each of seven patients, so far examined, a slope value and ST segment depression of greater than 1 mm were obtained which, according to previous experience, indicated myocardial ischemia equivalent to coronary heart disease. After aortic valve replacement, there was a significant reduction in heart size as assessed using the cardiothoracic ratio, and the amplitude of QRS complex on the electrocardiogram. The slope was abolished (two patients) or markedly reduced (five patients), and the decrease in the seven patients was statistically significant. ST segment depression could be obtained in one patient. This study has shown the occurrence of maximal ST/HR slope in patients without large coronary artery disease who have aortic valve disease and cardiac enlargement. PMID- 2965204 TI - Electrocardiographic evaluation of collateral development in conscious dogs. AB - In seven conscious dogs, endocardial ST-segment changes within the central ischemic area at 2 min after coronary occlusion were compared with the amount of reactive hyperemia and collateral blood flow. With the use of ultrasonic dimension gauges implanted in the subendocardium perfused by the left anterior descending coronary artery (LAD) and left circumflex coronary artery (LCCA), endocardial electrocardiograms and regional myocardial dimensions were simultaneously measured. Collateral vessels were developed by repeated 2 min LCCA occlusions during three weeks. Blood flow debt repayment following the release of LCCA occlusion was measured using a Doppler flowmeter. The collateral blood flow from LCCA to the area supplied by the occluded LAD was measured as a stepwise reduction in LCCA flow upon the release of LAD occlusion. With the attenuation of myocardial ischemia due to the collateral development, endocardial ST-segment shift revealed earlier restoration compared with subendocardial function. In the presence of mild ischemia, the ST-segment was still elevated. Thus, endocardial ST-segment changes serve as an indirect functional index of collateral development as well as regional myocardial function and blood flow debt repayment. PMID- 2965205 TI - Dopamine stimulates release of thyrotrophin-releasing hormone from perfused intact rat hypothalamus via hypothalamic D2-receptors. AB - We have studied the effect of dopamine together with agonist and antagonist drugs of different specificities on the release of TRH from the perfused, intact hypothalamus of the adult rat in vitro. Dopamine produced a dose-related stimulatory effect on TRH release with maximal effect being achieved at 1 mumol/l (increase over basal, 118 +/- 16.5 (S.E.M.) fmol TRH; P less than 0.001 vs basal). This effect was mimicked by the specific D2-agonist drugs bromocriptine (0.1 mumol/l) and LY 171555 (0.1 mumol/l) (increase over basal values, 137.5 +/- 13.75 fmol and 158.6 +/- 10.7 fmol respectively; P less than 0.001 vs basal), but not by the D1-agonist SKF 38393A. The stimulatory effect of dopamine (1 mumol/l) was blocked in a stereospecific manner by the active (D) but not by the inactive (L) isomers of the dopamine antagonist butaclamol. Similar blockade was achieved with the specific D2-antagonist domperidone (0.01 mumol/l) whereas the D1 antagonist SCH 23390 was only effective when used at a concentration 100 times greater. Lower concentrations (0.01 mumol/l) of this D1-antagonist did not block the stimulatory effect of dopamine. High-performance liquid chromatography characterization of the material secreted within the hypothalamus showed one single peak of immunoreactive material which coeluted with synthetic TRH. These data suggest that dopamine exerts a stimulatory role in the control of hypothalamic TRH release by acting at specific D2-receptors. PMID- 2965206 TI - Opioid peptides do not modulate atrial natriuretic peptide or aldosterone release under basal conditions in man. AB - The effect of two analogues of [Met]-enkephalin, [D-Ala2,N-Phe4,Met(0)-ol5] enkephalin and its guanyl derivative, on plasma concentrations of atrial natriuretic peptide (ANP) and serum aldosterone in six normal subjects was investigated. All subjects were given a 1 litre water load to inhibit vasopressin release. Both analogues, when injected i.v. at a dose of 100 micrograms, stimulated release of prolactin and GH and inhibited serum cortisol; there was no significant change in blood pressure, pulse rate or urine output. Neither plasma concentrations of ANP nor serum aldosterone levels changed significantly after injection of either analogue at a low or high dose. Naloxone, given i.v. as an 8 mg bolus, also failed to alter concentrations of either ANP or aldosterone, while it significantly stimulated the release of serum LH and cortisol. It was concluded that under basal conditions opiate receptors are unable to modulate plasma ANP or serum aldosterone concentrations. PMID- 2965207 TI - Lumbar disc syndrome in Finland. AB - The prevalence of lumbar disc syndrome (herniated disc or typical sciatica) and its consequences in terms of disability, handicap, and need for medical care were studied as part of the Mini-Finland Health Survey. A sample of 8000 persons representative of the Finnish population aged 30 or over was asked to come for examination, and 7217 (90%) participated. A diagnosis of lumbar disc syndrome based on medical history, symptoms, and standardised physical examination was made for 5.1% of the men and for 3.7% of the women. Half of these patients were assessed to be in need of medical care, over 80% of which was considered to be adequately met. One third of all patients with lumbar disc syndrome had been previously hospitalised for that syndrome, and one fifth of the patients had undergone lumbar surgery. At least slight disability was found in almost 60% of the patients, though severe functional limitations were rare. About 6% of the population's work disability was estimated to be attributable to lumbar disc syndrome. PMID- 2965208 TI - Agonist and antagonist effects of interferon alpha and beta on activation of human macrophages. Two classes of interferon gamma receptors and blockade of the high-affinity sites by interferon alpha or beta. AB - H2O2-releasing capacity and limited antitoxoplasma activity could be induced in human macrophages (derived from monocytes cultured greater than or equal to 5 d) but not in monocytes themselves (cells cultured less than or equal to 4 d) by a further 3-d incubation with pure natural or rIFN-alpha or -beta. More than 3 pM (10 U/ml) of these IFNs was required, with greatest effects at approximately 300 pM (10(3) U/ml). At 300 pM, H2O2-releasing capacity was enhanced 4.4 +/- 1.6-fold over medium control (mean +/- SD for natural INF-alpha, rIFN-alpha A, rIFN-alpha D, and rIFN-beta) compared to an 8.4 +/- 4.8-fold increase with rIFN-gamma (100 pM, 100 U/ml) in the same experiments. Unexpectedly, low concentrations of IFN alpha or -beta (3 fM-300 pM) blocked induction of H2O2-releasing capacity by rIFN gamma (10 pM), with a 50% inhibitory dose of approximately 80 fM. However, IFN alpha or -beta (3 fM-300 pM) could not inhibit the effect of higher concentrations of rIFN-gamma (1 nM). In contrast to results with monocytes or young macrophages, Scatchard plots of binding of 125I-rIFN-gamma to mature macrophages (day 8 of culture) indicated two classes of binding sites: approximately 2,000 high-affinity sites (Kd approximately 0.43 nM) and approximately 23,000 low-affinity sites (Kd approximately 6.4 nM) per cell. Binding of 125I-rIFN-gamma to the high- but not the low-affinity sites was blocked by simultaneously added IFN-alpha or -beta, with a 50% inhibitory dose of approximately 2 U/0.25 ml (approximately 2 pM), or reversed by subsequently added IFN-alpha or -beta. Thus, differentiation of human mononuclear phagocytes in vitro is accompanied by the emergence of (a) an agonist response to submicromolar concentrations of IFN-alpha or -beta, (b) antagonism of the effect of picomolar IFN-gamma by femtomolar IFN-alpha or -beta, (c) two classes of IFN-gamma-Rs, and (d) nonstimulatory binding of IFN-alpha or -beta to the high- but not the low affinity IFN-gamma-Rs, with higher affinity than rIFN-gamma itself. We speculate that traces of IFN-alpha or -beta derived from stromal cells, parenchymal cells, or resident macrophages may dampen the activation of mature tissue macrophages by the small amounts of IFN-gamma that diffuse from inflammatory sites into normal tissues. Such a mechanism could constrain the potentially destructive phenomenon of macrophage activation to areas where monocytes have recently immigrated and/or the concentration of IFNs is high. PMID- 2965209 TI - Fibronectin receptors of phagocytes. Characterization of the Arg-Gly-Asp binding proteins of human monocytes and polymorphonuclear leukocytes. AB - We have defined the cell surface molecules of human monocytes and PMN that bind to the chymotryptic cell binding domain of Fn and to a synthetic peptide, KYAVTGRGDS, based on the sequence of Fn, by affinity chromatography. Monocytes express two receptors that differ in their affinity for CBD-Sepharose and peptide Sepharose, but that both recognize the RGD sequence. Only a single receptor is purified from PMN, which resembles the monocyte surface molecule that binds to peptide-Sepharose. These receptors are not part of the Mac-1, LFA-1, p(150,95) family, but do have homology to the platelet Fn receptor, gpIIb/IIIa. Interestingly, the antigenic crossreactivity between gpIIb/IIIa and the phagocyte receptors purified on peptide-Sepharose is largely in the beta chain of the receptors. The alpha chains appear to be distinct, based on molecular weight, antigenic analysis, and ligand specificity. This receptor also seems to be the surface molecule on monocytes that is critical for phagocytosis enhancement by Fn. Thus, we have defined the phagocyte Fn receptor that transduces the signal for increased phagocytosis by monocytes; it may be a third member of a family of adhesion molecules that includes the gpIIb/IIIa of platelets and the vitronectin receptor of fibroblasts. PMID- 2965210 TI - Possible role of V beta T cell receptor genes in susceptibility to collagen induced arthritis in mice. AB - Arthritis was induced by immunization of type II collagen in adjuvant in mice from H-2q-bearing crosses between SWR (H-2q/q) and B10 (H-2b/b mice), two strains known to be resistant to collagen-induced arthritis (CIA). The resistance of B10 is known to be due to its MHC haplotype, but it was postulated that the resistance of SWR mice which expresses the susceptible MHC haplotype could be due to the deletion of close to 50% of the V beta genes of the T cell receptor (TCR) in them. 17% of the F1 hybrids, 33% of the SWR backcrosses, 68% of the B10 backcrosses, and 52% of the F2 hybrids developed arthritis on follow-up to 5 mo after primary immunization with collagen. There was no significant difference in anti-type II collagen antibody titers between the arthritic and nonarthritic mice in each of these crosses. The segregation of the TCR genes with arthritis was determined in the F2 population by typing with F23.1 mAb that reacts with T cells using V beta 8 subfamily genes in their TCRs. SWR mice are F23.1- as V beta 8 genes are deleted in them. All six of arthritic mice homozygous for H-2q, and thus with an H-2 haplotype similar to SWR mice, expressed the F23.1 marker. These studies indicate that for complete susceptibility to collagen-induced arthritis, not only is a susceptible MHC haplotype (H-2q) important, but possibly also the presence of a subset of T cells using certain specific V beta genes in their TCRs. Other background genes may, however, modulate the severity of arthritis. PMID- 2965213 TI - Electrophysiological study on hypertrophied and ischemic myocardial of rats. PMID- 2965211 TI - Induction of high-affinity interleukin 1 receptor on human peripheral blood lymphocytes by glucocorticoid hormones. AB - The in vitro effect of glucocorticoids (GCs) on IL-1-R expression of human PBMCs was investigated. Both physiological and pharmacological concentration ranges of GC increased the specific binding of 125I-labeled human rIL-1 alpha to PBMCs. This enhancement was specific for GC, since other steroid hormones, such as progesterone, 17 beta-estradiol, and testosterone failed to elevate the binding of 125I-IL-1 alpha to PBMCs. The effect was time dependent with maximal effect occurring 6 h after treatment and dose dependent with half-maximal effect elicited by 100 nM prednisolone. Scatchard plot analysis indicated that 125I-IL-1 alpha binding increased from approximately 100 IL-1-R per cell to 2 X 10(3) receptors per cell without a major change in affinity (Kd = 2.6 X 10(-10) M). The subpopulation of PBMCs induced by GC to express higher levels of IL-1-R consisted predominantly of B lymphocytes, but not T lymphocytes, large granular lymphocytes, or monocytes. GCs also induced the expression of IL-1-R on some other cell types, including normal human dermal fibroblasts and the human large granular lymphocyte cell line YT. Since cycloheximide and actinomycin D inhibited the induction of IL-1-R by GC, synthesis of both new RNA and protein seems to be required for IL-1-R induction. This study presents the first evidence of upregulation of the receptors for IL-1 by GC, and may account for the reported enhancement of in vitro and in vivo humoral immune responses by GCs. PMID- 2965212 TI - Production of tumor necrosis factor/cachectin by human T cell lines and peripheral blood T lymphocytes stimulated by phorbol myristate acetate and anti CD3 antibody. AB - The induction of mRNA synthesis and accumulation of TNF/cachectin and lymphotoxin (LT) mRNAs in T leukemic cell lines and freshly isolated T cells were studied by Northern blot analyses. Without stimulation, TNF mRNA was barely detected in four T cell lines (CEM, KE4, MT-1, and SKW-3) and not detectable in Molt-4 and Jurkat cells, while a considerable amount of TNF mRNA was observed in HSB-2 cells. When stimulated by PMA, these T cell lines accumulated varying levels of TNF mRNA. All seven T cell lines expressed LT mRNA when unstimulated and responded well to PMA by increased accumulation of LT mRNA. The calcium ionophore A23187 by itself had no effect on TNF and LT mRNA accumulations in these cell lines. The CD3+ T cell lines did not respond to anti-CD3 mAb T3-II alone. However, A23187 and mAb T3-II further elevated TNF and LT mRNA accumulations in PMA-treated T cell lines. Synergism between PMA and mAb T3-II was modest in the CD3+ cell lines. A slight difference in kinetics of TNF and LT mRNA accumulations was noted. In addition, heterogeneities in TNF and LT expressions by these cell lines in responses to PMA and other stimuli were observed. In monocyte-depleted peripheral blood T cell populations. PMA was able to induce both TNF and LT mRNA syntheses. This effect was potentiated markedly by the addition of anti-CD3 mAb T3-II. This synergistic response to anti-CD3 mAb and PMA provided further evidence that T cells were the source of TNF synthesis in these cultures. There was a difference in the kinetics of TNF mRNA accumulation and that of LT mRNA. Maximal accumulation of TNF mRNA occurred at 4 h while 8-18 h was required for maximal LT mRNA accumulation. IL-2 mRNA accumulated at an intermediate peak time of 4-8 h. Western blot analyses and cytotoxicity assays with L cells as targets indicated that these T cell lines and peripheral blood T cells secreted TNF. These results provide further evidence that human T cells are capable of making TNF as well as LT under appropriate stimulations. Their productions are an integral part of T cell response to activation signals. In addition, it appears that the production of these two closely related molecules is independently regulated. PMID- 2965214 TI - Psychopharmacological therapy of deviant sexual behavior. AB - Psychopharmacological approaches to controlling male deviant sexual behavior, especially sexual recidivism and sexual deviants on probation, have been reported in psychiatric literature. In Europe, the drug cyproterone acetate, and in the United States, medroxyprogesterone acetate, Provera, and in the long-acting form, Depo-Provera, have all benefitted exhibitionists and pedophiliacs, and reduced sex drive in sexual deviants. The combination of pharmacotherapy and either psychotherapy or behavioral therapy has been the most effective approach to reducing the sex drive of sexual deviants. PMID- 2965215 TI - Agent Orange exposure and posttraumatic stress disorder. AB - Evidence of organic psychological deficits in Vietnam veterans exposed to the herbicide Agent Orange was established through a neuropsychological battery. Also, the exposed Vietnam veterans, in contrast to a matched control group of Vietnam veterans, showed a significantly higher rate of posttraumatic stress disorder and its associated features: depression, anxiety, and increased aggression. The latter was subdivided into uncontrollable pressures, verbal violence, violence against objects, assaults, and suicidal thoughts. Active cases of chloracne, a medical indicator, were used to determine Agent Orange exposure. PMID- 2965216 TI - Persistent denial of handicap after infarction of the right basal ganglia: a case study. AB - A case is reported of persistent denial of handicap following stroke. Hemiplegia was due to infarction involving only sub-cortical structures, and there was no associated visual or sensory neglect or inattention, and no evidence of dementia. PMID- 2965217 TI - Determination of activated lymphocytes in peripheral blood of patients with multiple sclerosis. AB - Using monoclonal antibodies Ta1 and CD25 (interleukin-2 receptor: I-2R) and flow cytometry, the levels of activated lymphocytes in the peripheral blood of 50 patients with multiple sclerosis (16 relapsing inactive; 14 relapsing active; 20 chronic progressive) and 20 normal subjects were investigated. No significant differences were observed in the percentage or absolute numbers of Ta1 and IL-2R reactive lymphocytes between the normal and multiple sclerosis patient groups, irrespective of disease activity. Monitoring peripheral blood lymphocytes with respect to these markers would appear to have little value in the management of multiple sclerosis. PMID- 2965218 TI - Prefrontal cortical blood flow and cognitive function in Huntington's disease. AB - To examine the relationship between cortical physiology and dementia in Huntington's disease, rCBF during three different behavioural conditions, one of which emphasised prefrontal cognition, was determined by xenon-133 inhalation in 14 patients with Huntington's disease and in matched controls. Cortical rCBF was not reduced in Huntington's disease patients even while they manifested overt prefrontal-type cognitive deficits. Caudate atrophy on CT and rCBF were significantly correlated, but only during the prefrontal behaviour where the correlation was positive. These results suggest a qualification of the subcortical dementia concept as applied to Huntington's disease and implicate an interaction between pathology that is subcortical and cognitive function that is cortical. PMID- 2965219 TI - Effectiveness and tolerability of low-dose cyclophosphamide and low-dose intravenous interleukin-2 in disseminated melanoma [corrected]. AB - We studied the effects on melanoma of low-dose recombinant interleukin-2 (IL-2) preceded by low-dose cyclophosphamide (CYC). Twenty-seven outpatients, aged 25 to 75 years, were treated with IL-2, 3.6 million U/m2 intravenously (IV), daily for five days on 2 successive weeks beginning three days after 350 mg/m2 of IV CYC. This schedule was repeated at least twice more at 1-week intervals. Six of 24 patients (25%) who received more than one 2-week cycle of treatment had a remission, one complete and five partial, with minor responses in eight others (33.3%). Three patients with rapidly progressive disease, who received only one cycle, were excluded from the analysis of response. The responses comprised remissions of liver metastases in two patients, one of them complete, two complete and two partial regressions of subcutaneous metastases, partial remission of lymph node metastases, and a partial remission of lung nodules. The mean duration of response exceeded 5 months, with two patients treated for greater than 1 year. Toxicity was moderate and controllable and only two patients required hospitalization, both overnight. Lymphokine-activated killer (LAK) cell activation was induced in 17 of the 24 patients, including all six responders, while none of seven patients without LAK activation had a remission. This regimen appeared to be as effective in melanoma as those involving ex vivo activation of LAK cells, and was generally tolerable to patients in all age groups. PMID- 2965220 TI - Effects of short-term dehydroepiandrosterone treatment on serum and pancreatic insulin in Zucker rats. AB - Lean and obese female Zucker rats were treated with 0.6% dehydroepiandrosterone (DHEA) from 5 until 10 wk of age and comparisons made to both ad libitum--fed and weight-matched groups. Within genotype, body weights and cumulative food intakes of DHEA and weight-matched groups were not different. Fat pad weights of DHEA rats were significantly lower than those of non-treated groups. Pancreatic insulin content, whether expressed per pancreas or per gram pancreas, was not lowered by DHEA treatment. Fasting serum insulin levels were not altered in lean rats but were significantly lower in DHEA obese rats than in either nontreated obese group. Glucose metabolism (conversion to CO2, fatty acids or glyceride glycerol) in isolated adipocytes was similar in all groups except that obese rats had greater glyceride-glycerol production than lean rats. Glucose conversion to CO2 in soleus muscle was lower in obese rats than in lean rats. Basal and insulin stimulated glycogen production was lower in DHEA and weight-matched obese rats than in any other group. Ad libitum-fed lean rats had higher insulin-stimulated glycogen production than DHEA lean and all groups of obese rats. Short-term DHEA treatment of obese Zucker rats lowers serum insulin levels without improvement of insulin resistance in peripheral tissues and without lowering pancreatic insulin content. PMID- 2965221 TI - Work status and response to epidural steroid injection. PMID- 2965222 TI - US Navy shipyard occupational medicine through World War II. AB - For more than 60 years the US Navy has maintained occupational health programs for its civil service workers in shipyards, arsenals, and aircraft repair facilities. The early history of the organization, people, and professional activities dedicated to the health of this large federal industrial workforce is examined. Early efforts were stimulated by increasingly complex naval technology and worker compensation law. During World War II training, clinical, and preventive programs were pursued vigorously. Navy occupational health paralleled and at times led the development of occupational medicine and industrial hygiene in America. PMID- 2965223 TI - C. O. Sappington memorial lecture. The vanguard and the rearguard: occupational medicine revisits its future. PMID- 2965224 TI - A comparison between the Minnesota Multiphasic Personality Inventory and the 'Mensana Clinic Back Pain Test' for validating the complaint of chronic back pain. AB - Reports on the efficacy of the Minnesota Multiphasic Personality Inventory (MMPI) for selecting patients with valid complaints of pain have been equivocal. The Mensana Clinic Back Pain Test (MPT) was able to predict, with some degree of success, patients who had a definite organic pathologic condition. However, the MMPI measures personality traits, whereas the MPT measures the impact of pain on a patient's life. To determine which of the two tests would be a better predictor of actual physical abnormalities, and hence valid pain complaints, a comparison was undertaken between the two tests. The charts of 83 patients admitted to the Neurosurgery Service of Johns Hopkins Hospital with complaints of back pain were assessed. MMPI test results, as well as test results for the MPT, were compared to the presence or absence of pathologic conditions on electromyography, nerve conduction velocity studies, thermography, myelogram, or computerized axial tomography scan. The MPT had a correlation factor of -.59700, that was significant as P = .000005. Of the 52 patients scoring 17 points or less on the MPT, 85% had objective physical abnormalities, considered moderate or severe by blind review. Of the 31 patients scoring 18 points or greater on the MPT, only 26% had objective physical findings that were considered moderate or severe. Only the F scale (faking badly) of the MMPI correlated with objective physical abnormalities (r = .21340, P less than .033). However, 60% of the patients with T scores of less than 70 on the F scale had objective findings, whereas 75% of patients with T scores greater than 70 had objective physical findings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965225 TI - Cardiovascular reactivity, plasma markers of endothelial and platelet activity and plasma renin activity after mental stress in normals and hypertensives. AB - Endothelial injury and platelet activation, mechanisms known to be involved in vascular lesions, may promote the development of cardiovascular disorders possibly associated with mental stress. Plasma markers of platelet activation (beta-thromboglobulin, BTG) and of endothelium activity (factor VIII/von Willebrand factor, FVIII/vWf) and plasma renin activity (PRA) were determined in 17 healthy normotensive volunteers and in 21 hypertensives without target-organ damage before and after mental stress (a colour-word conflict test). The aim of the study was to compare cardiovascular reactivity with the stress-induced changes in platelet and endothelium activity. Individual responses in BTG and factor FVIII/vWf after the colour-word conflict test were markedly different, but significant mean increases were observed in both groups with no difference in the degree of response and in the percentage of responders. No correlations were found among the changes in plasma variables or between cardiovascular reactivity (systolic and diastolic blood pressure and heart rate) and the changes in BTG, FVIII/vWf and PRA. These findings suggest that hypertensive patients do not have an abnormal platelet or endothelium reactivity to mental stress, at least when the disease is free of vascular complications. This dissociation of stress induced variability in BTG and FVIII/vWf and cardiovascular reactivity indicates that these indices could be used as independent markers of mental stress. PMID- 2965226 TI - Consequences of reversal of hypertensive cardiac hypertrophy by captopril on left ventricular pumping ability and performance. AB - Left ventricular hypertrophy can be reversed by treatment of hypertension with captopril but the consequences of this regression are not yet fully described. We studied the maximal capacity of the hypertrophied and hypertrophy-reversed ventricle to generate pressure during transient total occlusion of the aorta, and also the left ventricular end-diastolic pressure required to meet this maximal effort. Two-kidney, one clip Goldblatt (renal hypertensive rats; RHR) hypertension was induced in 17 Wistar rats, eight of which were treated with captopril (RHR-C: 50 mg/kg given orally) from the fourth to the eighth week. Sham operated controls (SC) remained untreated, or were treated with similar doses of captopril (SC-C). Significantly lower heart weights were found in RHR-C than in RHR (2.88 +/- 0.15 versus 2.38 +/- 0.04; P less than 0.001). During transient total occlusion of the aorta, the maximal intraventricular pressure developed in RHR-C was not significantly different from that in RHR, but left ventricular end diastolic pressure was significantly less in RHR-C than in RHR (21.4 +/- 2.2 versus 34.3 +/- 3.8; P less than 0.01). The analysis of pressure-volume characteristics of the hypertrophied left ventricles and those in which hypertrophy was reversed revealed similar compliances between these two groups. Our data suggest that there was a mechanical improvement in the heart function after reversal of left ventricular hypertrophy. PMID- 2965227 TI - Heart and vessel hypertrophy in hypertension: possibilities of regression. AB - Fifty-two middle-aged hypertensive males with left ventricular hypertrophy, diagnosed by echocardiography, were divided into two groups; 30 of them were treated with beta-blockers and 22 with methyldopa. Where blood pressure was insufficiently controlled, diuretics and, eventually vasodilating agents, were added to both treatment regimes. During 2 years of follow-up, blood pressure decreased significantly. The greatest decrease was observed during the first 3 months. Echocardiography revealed the greatest decrease in posterior wall and interventricular septal thickness also during the first 3 months. Posterior wall hypertrophy was reversed completely in the next 3 months, while further regression of interventricular septal hypertrophy was continuous during the remaining follow-up period. Left ventricular hypertrophy was reversed completely in 27 (51.9%) probands and remained unchanged in two patients. The regression was incomplete in 23 (44.2%) patients who were more obese and whose left ventricular hypertrophy was initially more pronounced. Regression of hypertrophy was not associated with a deterioration in left ventricular function and was achieved even after the addition of vasodilating drugs. No differences were observed between the treatment groups. The pathogenesis of left ventricular hypertrophy still remains unclear. After 2 years, no changes in peripheral vascular resistance in the forearm were observed at reactive hyperaemia. These results indicate that there might be a delay in the regression of vessel hypertrophy compared with that of the left ventricle. PMID- 2965228 TI - Effect of regression of left ventricular hypertrophy following sotalol therapy on diastolic function in hypertensive patients. AB - The effects of changes in left ventricular mass following beta-adrenergic blockade therapy (sotalol) on left ventricular filling indices were examined in 16 patients with essential hypertension aged 46 +/- 8.3 years (mean +/- s.d.). The peak atrial to peak early diastolic velocity (A:E) ratio and the peak filling rate (PFR = peak early diastolic velocity X mitral annulus area) were measured by the use of pulsed Doppler left ventricular inflow time-velocity plots following placebo and after 8-12 weeks of sotalol monotherapy (160 mg/day). Compared with normal controls of similar age, our patients had a larger left ventricular mass and impaired left ventricular filling indices. Following sotalol, mean arterial pressure (MAP) decreased by 14%, the heart rate by 15% and left ventricular mass by 11%, while diastolic filling improved (A:E 17% and PFR 21%). Six patients had a decrease in left ventricular mass of greater than 15% (group A); other patients (group B) showed a smaller reduction. In spite of comparable changes in MAP and heart rate in groups A and B, the patients in group A showed a smaller degree of improvement in the A:E ratio. It is concluded that sotalol can induce regression of left ventricular hypertrophy and improve left ventricular filling in hypertensive patients, and it seems that improvement in diastolic filling is related to a reduction in afterload rather than to a decrease in left ventricular mass. PMID- 2965229 TI - Long-term effect of antihypertensive therapy on left ventricular hypertrophy. AB - We studied a total of 88 previously untreated hypertensives in order to determine the long-term effect of antihypertensive therapy on echocardiographically confirmed left ventricular hypertrophy. The drugs tested were the beta-blocker metoprolol (200 mg/day; 26 patients; mean age 43.9 years; follow-up 32 months), the calcium antagonist gallopamil (100-150 mg/day; 26 patients; mean age 49.7 years; follow-up 23 months) and a combination of 50 mg atenolol and 20 mg nifedipine (36 patients; mean age 44.2 years; follow-up 18 months). Despite similar reductions in resting blood pressure, each of the three therapeutical regimens had a different effect on the left ventricular mass index after 1 year of treatment: gallopamil reduced the index by 13.2% (170 +/- 49 to 148 +/- 41 g/m2, P less than 0.001), metoprolol by 21.9% (150 +/- 27 to 117 +/- 27 g/m2, P less than 0.001) and atenolol + nifedipine by 30.8% (148 +/- 33 to 103 +/- 21 g/m2, P less than 0.001). Similar results were obtained for interventricular septal (9.9, 15.4 and 20.7%, respectively) and posterior wall thicknesses (10.4, 16.7 and 24.1%, respectively). During the follow-up there was a further significant reduction in ventricular hypertrophy under all three treatments, but no significant changes in ventricular end-diastolic and end-systolic dimensions or in fractional shortening. PMID- 2965230 TI - Greater vasodilator responsiveness to atrial natriuretic peptide in low-renin essential hypertensives. AB - Forearm vasodilator responses to atrial natriuretic peptide (ANP) were studied in twelve untreated patients with essential hypertension and twelve normotensive subjects. Alpha-human ANP (0.005 to 1.5 micrograms/min per 100 ml forearm volume) infused into the brachial artery increased forearm blood flow dose-dependently. This was paralleled by a decrease in forearm vascular resistance (FVR) which, at lower doses, was greater in essential hypertensives than in normotensives (P less than 0.001), and showed a lower ED50 for ANP in essential hypertensives (P less than 0.01). At higher doses of ANP the difference in vasodilator response between hypertension and normotension disappeared; the response to ANP was associated with a fall (P less than 0.01) in systemic blood pressure in hypertensives but not normotensives. At lower doses, the decreases in FVR were correlated directly with plasma renin activity in hypertensives (r = 0.656; P less than 0.05) but not normotensives. These data suggest greater vasodilator responsiveness to infusions of low doses of ANP in essential hypertensives, which is greater in low-renin states and blunted in high-renin states. PMID- 2965231 TI - The long-term effect of isradipine in pindolol-treated patients. AB - The long-term efficacy of isradipine, a new dihydropyridine calcium antagonist with marked vascular selectivity, was evaluated in 17 patients with essential hypertension. All had a supine diastolic blood pressure of greater than 95 mmHg with 10 mg pindolol once daily. After a short-term, double-blind, dose-finding, crossover comparison with addition of isradipine or placebo twice daily, they continued on pindolol and their optimal dose of isradipine in a single-blind, long-term follow-up study. Eighteen patients were recruited but one male patient discontinued treatment after 2 weeks due to ankle oedema and will not be accounted for in the overall evaluation. There were 11 males and six females with a mean age of 56 +/- 10 years. In the short-term study on the optimal dose of isradipine (5.1 mg twice daily) blood pressure was lowered by 24/18 mmHg (P less than 0.001). No change in heart rate was seen despite the substantial drop in blood pressure. In the long-term study the patients were seen for a mean follow up time of 12.5 months (range 4-17 months). After the longest follow-up time mean arterial pressure was 107.0 +/- 7.4 compared with 120.1 +/- 8.2 mmHg after placebo baseline [delta = 13 mmHg (11%), P less than 0.001, n = 17]. The heart rate was unchanged (delta = 0.2 beats/min, 95% confidence limits -3, +3), and so was ankle circumference (delta = 0.12 cm, 95% confidence interval, -1, +1). On the other hand, mean weight was reduced by 2 kg from 90 kg (P less than 0.05, n = 17).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965232 TI - Renal effects of angiotensin II, atrial natriuretic peptide and their interaction in man. AB - There is now much evidence that atrial natriuretic peptide (ANP) is important in the control of sodium balance. There is also evidence that ANP interacts with the renin-angiotensin system at several levels. In this study we investigated a further possible interaction between ANP and angiotensin II (ANG II) in the control of renal water and electrolyte excretion. In normal male volunteers, ANP caused urinary sodium excretion to rise significantly from baseline (+80 +/- 44 mumol/min) whereas ANG II was potently antinatriuretic (-125 +/- 36 mumol/min). When ANP was administered against a nonpressor background infusion of ANG II, urinary sodium excretion rose from a new lower level (-93 +/- 22 mumol/min) to a rate not significantly different from control (-22 +/- 25 mumol/min). In contrast to that of sodium, potassium excretion and urinary volume remained significantly below baseline levels when ANP was administered against a background infusion of ANG II. These results suggest that ANP and ANG II interact within the kidney in the control of water and electrolyte excretion. PMID- 2965233 TI - Influence of atrial natriuretic factor on forearm reflex vasoconstriction induced by cardiopulmonary or arterial receptor unloading. AB - We studied the influence of atrial natriuretic factor (ANF) infusion on the reflex increase in forearm vascular resistance in normotensive subjects. Reflex vasoconstriction was induced by cardiopulmonary receptor unloading [lower body negative pressure (LBNP), -20 mmHg for 15 min] or by carotid baroreceptor deactivation (+60 mmHg increase in external neck pressure by a pneumatic neck chamber). Atrial natriuretic factor induced a significant increase in the reflex forearm vasoconstriction to LBNP, but did not modify systemic and regional reflex haemodynamic responses to carotid baroreceptor deactivation. These results suggest that ANF has important interactions with the neural control of peripheral circulation. In addition, the study shows that the peptide causes a selective potentiation of the reflex vasoconstrictor response evoked by cardiopulmonary receptor unloading. PMID- 2965234 TI - Physiological stimuli to atrial natriuretic peptide secretion in normal humans. AB - We tested the response of plasma atrial natriuretic peptide (ANP) levels to the following physiological stimuli: postural changes; head-out water immersion; and physical exercise. Plasma ANP (p-ANP) levels were assessed by a specific, sensitive radio-immunoassay. Plasma ANP rose significantly when posture shifted from upright to recumbent for 1 h, but fell again to basal values after 10 min standing. Circadian variations did not affect the posture study. Head-out water immersion produced a prompt and remarkable (sevenfold) increase in p-ANP, with a plateau reached after 1 h and held until the end of the experiment (2 h). Plasma ANP levels were measured in 10 normal subjects performing supine treadmill exercise at 50% of maximum aerobic capacity for 30 min. Plasma ANP rose from baseline supine values after 15 min exercise, and remained elevated during the following 15 min exercise. During the recovery phase ANP showed a trend towards baseline values, with a 38% decrease attained after 30 min. We propose that the above tests could be used as ANP-stimulating manoeuvres in physiological and clinical conditions in man. PMID- 2965235 TI - Atrial natriuretic peptide levels during angiotensin infusion and indomethacin administration are consistent with angiotensin-mediated regulation in man. AB - Low-dose angiotensin II (ANG II) infusion raised plasma atrial natriuretic peptide (ANP) levels only when endogenous renin-angiotensin levels were low, as in aldosterone-producing adenoma. When plasma renin activity (PRA) levels rose tenfold following removal of the tumour, low-dose ANG II infusion no longer stimulated ANP, but fivefold higher doses did. Indomethacin lowered both PRA and ANP in Bartter's syndrome and in normal subjects. The effect of indomethacin on ANP is probably not direct, since it did not lower ANP in aldosterone-producing adenoma. Neither did it lower PRA in aldosterone-producing adenoma, and in most studies ANP and PRA moved in parallel, consistent with positive regulation of ANP by ANG II. When ANG II infusion stimulated ANP, it also raised blood pressure, which could therefore be mediating the effects of ANG II on ANP. However, both PRA and ANP are high in Bartter's syndrome, while blood pressure is normal or low, and indomethacin lowers PRA and ANP in Bartter's syndrome and in normal subjects without lowering the blood pressure. The relative importance of regulatory factors such as central blood volume/atrial pressure and ANG II level probably varies in different situations. In aldosterone-producing adenoma, a high central blood volume appears to over-ride the effect of a low ANG II level. In Bartter's syndrome a high ANG II level appears to over-ride the effect of low central blood volume. PMID- 2965236 TI - Drug abuse in the Georgian S.S.R. PMID- 2965237 TI - Pachyonychia congenita in an 11 year old female. PMID- 2965238 TI - Behavior management of the pediatric cancer patient. PMID- 2965239 TI - The dentinal surface of composite restorations after chemo-mechanical caries removal. PMID- 2965240 TI - Dental management of patients with Russell-Silver syndrome. PMID- 2965241 TI - Disability from endometriosis in the United States Army. AB - The impact of endometriosis on the general population has been well documented, and the prevalence rate is quoted as 10%. The records of 6,456 women in the United States Army were reviewed with respect to age, race and active duty status. The active duty records were further evaluated for total sick days from endometriosis-related problems. This evaluation revealed a mean sick time of 15 days per admission per active-duty patient regardless of age or race. The estimate of lost duty time was 21,746 days, considering all active-duty patients in the study (five years of records). The prevalence of endometriosis in the female U.S. Army population was 6.2%. Diagnosis and treatment of endometriosis should be aggressively pursued whenever the disease is suspected. PMID- 2965242 TI - Pustulotic arthroosteitis and chronic recurrent multifocal osteomyelitis in children. Report of three cases. AB - We describe 3 children with pustulosis palmaris et plantaris associated with chronic recurrent (multifocal) osteomyelitis, located in the clavicle in 2 patients, and in the distal fibula as well as in several metatarsals in the third. The first 2 children also developed inflammatory sacroiliitis, one with chronic peripheral polyarthritis. These 3 cases represent the arthrosteitis and chronic recurrent multifocal osteomyelitis syndrome associated with palmoplantar pustulosis. PMID- 2965243 TI - Design, synthesis, and biological activity of a peptide mimic of vasopressin. AB - Our molecular modeling studies suggested that the conformational effects of the "cystine-line" residue Pmp1-Cys6 on the cyclohexapeptide ring of the vasopressin antagonist [Pmp1,D-Phe2,Val4]AVP might be mimicked by substitution of D aminoadipic acid at position 6 and cyclization of its side-chain carboxyl to the alpha-amine of residue 2. The peptide was prepared with DL-aminoadipic acid, and following cyclization, the two diastereomeric peptides were separated and purified by preparative high-performance liquid chromatography. The structure of each was confirmed by amino acid analysis and fast atom bombardment mass spectrometry. The chirality of the aminoadipic acid residue of each peptide was determined by chiral gas chromatography. The circular dichroism spectrum of each peptide was run and compared with the appropriate agonist and antagonist peptide standards. These peptides demonstrated in vitro poor V2-receptor affinity and an inability to inhibit or stimulate vasopressin-induced adenylate cyclase formation, suggesting that they lack one or more key features of the agonist/antagonist pharmacophore. PMID- 2965244 TI - N,N-di-n-propylserotonin: binding at serotonin binding sites and a comparison with 8-hydroxy-2-(di-n-propylamino)tetralin. AB - 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) is a serotonergic agonist with high affinity and selectivity for a particular population of central serotonin (5 HT) binding sites (i.e., 5-HT1A sites). Because the selectivity of 8-OH-DPAT may be due to the terminal amine substituents, the di-n-propyl analogue of 5-HT (i.e., 4) and of 5-methoxytryptamine (i.e., 5) were prepared and compared with 8 OH-DPAT with respect to their binding profile. Unlike 8-OH-DPAT, neither compound 4 nor 5 displays selectivity for 5-HT1A vs 5-HT2 sites. Consistent with these results, stimulus generalization occurs with 5 both in rats trained to discriminate 8-OH-DPAT from saline and in rats trained to discriminate the 5-HT2 agonist DOM from saline. The results of this study suggest that it is not the N,N dipropyl groups that account for selectivity, but, rather, it is some feature associated with the pyrrole portion of the indolylalkanamines that is important. PMID- 2965245 TI - Treatment of defective newborns--a survey of paediatricians in Poland. AB - We report the results of a survey of the attitudes and practices of paediatricians in Warsaw, Poland, with respect to the treatment of infants born with severe handicaps. The results are compared with a similar survey conducted by Australian researchers (1). In the Polish medical community surveyed, unconditional respect for life is a dominant attitude. Our study has revealed a deeply-entrenched paternalistic attitude among Polish doctors and a strong unwillingness to distinguish between 'ordinary and extraordinary means' of prolonging life, as well as an ambivalent attitude towards legal regulations binding in Poland. PMID- 2965246 TI - Handicapped infants: medical ethics and the law. AB - The main purpose of this paper (1) is to draw attention to a gap between the principles of Common Law and the principles accepted by many leading medical practitioners on the ethics of allowing severely handicapped infants to die. The Common Law principles are shown in Court of Appeal judgements on two cases. The contrasting principles of many paediatricians were illustrated at the trial of Dr Leonard Arthur. The paper suggests that the gap could be closed by statutory guidance on general principles. It also argues that utilitarian concepts misrepresent the ethical issues of medical dilemmas like this one. PMID- 2965247 TI - Care of the severely handicapped. PMID- 2965248 TI - Prediction of an ATP reactive center in the small subunit, gpNu1, of the phage lambda terminase enzyme. AB - The small subunit of the bacteriophage lambda terminase enzyme, the product of the phage's Nu1 gene, is shown to contain amino acid segments homologous to those present in a large number of ATPases. In keeping with these predictions, the purified protein has been found to hydrolyze ATP with a relatively low turnover number. Terminase holoenzyme is a known ATPase, and the biochemical significance of an ATP-interactive center situated in the gpNu1 subunit is discussed. PMID- 2965249 TI - Dominance in lambda S mutations and evidence for translational control. AB - Phenotypic analysis of a collection of point mutations in the lysis gene S of bacteriophage lambda indicates that many of the S alleles exhibit at least partially dominant character, suggesting that the S gene product (gpS) must oligomerize to achieve its lethal membrane effect. Moreover, mutations found 5' to the coding sequence also show a dominant character and appear to define a site, designated sdi (structure directed initiation) where mRNA secondary structure controls the choice of initiation codons. We propose that formation of the sdi structure occludes the consensus Shine-Dalgarno sequence and results in initiation at the Met3 codon, generating a lethal 105 residue polypeptide. The model predicts that, in the absence of the sdi stem-and-loop, initiation occurs at the Met1 codon, generating a 107 residue polypeptide, which is a non-lethal inhibitor of lysis. In support of the model, alteration of the first codon was achieved using site-directed mutagenesis, resulting in an S allele that is more lethal and induces lysis significantly sooner than the wild-type. PMID- 2965250 TI - Secondary structure and thermostability of the phage P22 tailspike. XX. Analysis by Raman spectroscopy of the wild-type protein and a temperature-sensitive folding mutant. AB - The thermostable tailspike endorhamnosidase of bacteriophage P22 has been investigated by laser Raman spectroscopy to determine the protein's secondary structure and the basis of its thermostability. The conformation of the native tailspike, determined by Raman amide I and amide III band analyses, is 52 to 61% beta-sheet, 24 to 27% alpha-helix, 15 to 21% beta-turn and 0 to 10% other structure types. The secondary structure of the wild-type tailspike, as monitored by the conformation-sensitive Raman amide bands, was stable to 80 degrees C, denatured reversibly between 80 and 90 degrees C, and irreversibly above 90 degrees C. The purified native form of a temperature-sensitive folding mutant (tsU38) contains secondary structures virtually identical to those in the wild type in aqueous solution at physiological conditions (0.05 M-Na+ (pH 7.5], at both permissive (20 degrees C) and restrictive (40 degrees C) temperatures. This supports previous results showing that the mutational defect at 40 degrees C affects intermediates in the folding pathway rather than the native structure. At temperatures above 60 degrees C the wild-type and mutant forms were distinguishable: the reversible and irreversible denaturation thresholds were approximately 15 to 20 degrees C lower in the mutant than in the wild-type protein. The irreversible denaturation of the mutant tailspikes led to different aggregation/polymerization products from the wild-type, indicating that the mutation altered the unfolding pathway. In both cases only a small percentage of the native secondary structure was altered by irreversible thermal denaturation, indicating that the aggregated states retain considerable native structure. PMID- 2965251 TI - Bacteriophage lambda DNA packaging. The product of the FI gene promotes the incorporation of the prohead to the DNA-terminase complex. AB - Lambda DNA packaging in vitro can be examined in stages. In a first step, lambda DNA interacts with terminase to form a DNA-enzyme complex, called complex I. Upon addition of proheads, in a second step, a ternary complex, complex II, containing DNA, terminase and the prohead is formed. Finally, upon addition of the rest of the morphogenetic components, complete phages are assembled. We have investigated the effect of the FI gene product (gpFI) in these reactions and found that a stimulation in phage yield is observed when gpFI is included early in the reaction, at the time when DNA, terminase and proheads interact to form complex II. Measurements of complex II formation revealed that gpFI stimulated the rate of formation of this intermediate. gpFI was further shown to stimulate the addition of proheads to preformed complexes I to give complex II, but the protein did not stimulate complex I formation. PMID- 2965252 TI - Growth of capillaries and myocardial cells in the normal rat heart. AB - The normal growth process of myocardial cells and capillaries in the late postnatal period (after weaning) was studied in 33 normal male Wistar rats (group I: 5 weeks, n = 9; group II: 7 weeks, n = 7; group III: 13 weeks, n = 8; group IV: 52 weeks, n = 9). The rats were fixed by retrograde vascular perfusion via the abdominal aorta. Two transverse and 2 longitudinal sections per animal were selected at random from the left ventricular papillary muscles for light and electron microscopic stereological investigation. Length density and surface density of myocardial cells and capillaries were estimated with correction for partial anisotropy and curvature by means of the mathematical model of a Dimroth Watson orientation distribution. The results were analyzed by allometric techniques. Relative left and right ventricular weight decreased continuously throughout the growth process; in terms of allometry, ventricular weight was proportional to (body weight). The capillaries showed predominantly longitudinal growth with nearly constant cross-sectional area, whereas the myocardial cells grew by nearly harmonic enlargement into all three directions of space. The relation between total capillary length and left ventricular weight could be described in close approximation by the allometric model (r = 0.98, P less than 0.001). Corresponding to the result that total capillary length was proportional to (left ventricular weight)0.71, the myocardial capillarization (length, surface area, and volume of capillaries per unit tissue volume) decreased with increasing heart size. As allometry makes possible the quantitation of the intensity of growth processes by a single dimensionless number (the allometric exponent), this method permits a convenient interlaboratory comparison of growth studies, and it allows a comparison of capillary reactions during maturation and in experimental cardiac hypertrophy. PMID- 2965253 TI - Comparison of human erythrocyte insulin binding and adenosinetriphosphatase activity. AB - Twelve healthy, non-obese male volunteers were selected to measure cellular magnesium (Mg(++)), calcium (Ca(++)), sodium/potassium (Na(+)/K(+)), and adenosinetriphosphatase (ATPase) activities. Measurements were performed using a crude hemolysate as well as a membrane fraction representing cytosolic pump activity. Binding-site data were subjected to Scatchard analysis for determination of receptor number and affinity.There were 432 +/- 1.87 insulin binding sites per cell. ATPase activity was measured in units of moles of inorganic phosphate (Pi) released per gram of hemoglobin (Hgb) every two hours (MUmol of Pi/g of Hgb/2 h). Na(+)/K(+)-ATPase activity in the hemolysate and membrane fractions was 18 +/- 0.99 MUmol of Pi/g of Hgb/2 h and 11.0 +/- 0.88 MUmol of Pi/g of Hgb/2 h, respectively. Calcium-ATPase activity was 136 +/- 1.92 MUmol of Pi/g of Hgb/2 h in the hemolysate, and 82 +/- 2.07 MUmol of Pi/g of Hgb/2 h in the membrane. Magnesium-ATPase activity in these fractions measured 24 +/- 1.09 and 16 +/- 0.91, respectively.Linear regression analysis of binding capability vs pump activity failed to disclose a significant relationship. Given the narrow range of values in this small study group, a much larger sample size will be required before a relationship can be established. PMID- 2965254 TI - Performance tests of Doppler ultrasound equipment with a tissue and blood mimicking phantom. AB - A tissue- and blood-mimicking phantom was assembled for assessing the performance of ultrasound Doppler equipment. The phantom is in the shape of a rectangular parallelepiped with a 10 x 20 cm scanning window and a depth of 16 cm. Components of the phantom include a tissue-mimicking material, 7.9 mm diameter simulated vessels, a fluid with similar back-scatter as whole blood, and a peristaltic pumping system producing peak scatterer velocities greater than 1 m/sec. Performance tests done with the phantom are outlined. These include assessments of the maximum depth of penetration and of the directional discrimination capabilities of the instrument, determinations of the accuracy of displayed flow velocities, and accuracy assessments of the displayed position of the Doppler sample volume. PMID- 2965255 TI - Laser-induced fluorescence detection of atherosclerotic plaque with hematoporphyrin derivative used as an exogenous probe. AB - Clinical use of laser angioplasty is limited by the lack of an adequate guidance system. As a first step toward developing a reliable guidance system, laser induced surface fluorescence and a fluorescent probe were used to differentiate plaque from normal arterial wall. The aortas from four normal New Zealand white rabbits and six atherosclerotic rabbits were studied in vitro. Rabbits from each group received 2.5 mg/kg of hematoporphyrin derivative intravenously 24 hours before death. Segments of the aortas were irradiated with a helium-cadmium laser, and the tissue surface fluorescence spectra were recorded with an optical multichannel analyzer. A plaque index, based on the resulting spectra, was calculated for each specimen of aorta. The spectra from normal aorta without hematoporphyrin derivative, normal aorta with hematoporphyrin derivative, and from plaque of atherosclerotic rabbits without hematoporphyrin derivative showed the same wavelength dependence. The plaque index values were not significantly different from one another. However, in plaque from atherosclerotic rabbits given hematoporphyrin derivative, the spectrum was markedly different, showing a broad spectral peak at 632 nm. This spectral peak corresponds to the spectral peak of hematoporphyrin derivative and was only seen in the plaque of atherosclerotic rabbits given hematoporphyrin derivative. The plaque index for these specimens was significantly different from all other specimens (p less than 0.001). This difference in fluorescence spectra and plaque index could be incorporated into a guidance system for laser angioplasty. PMID- 2965256 TI - How should we preclot knitted Dacron grafts? AB - We studied the change in water porosity over time of 10 Sauvage Bionit-II and 10 DeBakey Vasculour-II knitted velour Dacron grafts throughout the four stages of the Sauvage preclotting technique. Graft porosity decreased significantly (p less than 0.001) at the ends of stages 1 and 2 for both types of grafts, but stages 3 and 4 did not further reduce graft porosity. These results demonstrate that a two stage preclotting technique is adequate for the preclotting of knitted velour Dacron grafts. A final rinse with heparinized blood is recommended as this may reduce graft thrombogenicity. PMID- 2965257 TI - Effects of argon laser on vascular materials. AB - The laser has been shown to vaporize arterial atheromatous plaque, but little information is available about its effect on various vascular materials. This study attempts to determine a safe margin for laser graft thrombectomy. An 18 watt argon laser with a 2 mm metal-encased sapphire tip fiberoptic system was used. Specimens such as autogenous vein, polytetrafluoroethylene (PTFE), and Dacron grafts as well as polypropylene and PTFE sutures were harvested from amputated human legs. Laser energy was delivered from the tip of the fiberoptic system while perpendicular to and in direct contact with these materials. The effects were examined histologically and compared by determining the bursting pressure of the grafts and the breaking strength of the sutures. The results indicate that seven sections of autogenous vein grafts and 11 sections of PTFE grafts were perforated by laser delivery of 32.9 +/- 9.9 and 50.0 +/- 7.1 joules, respectively, whereas two sections of Dacron grafts were easily perforated after only 10 joules was delivered. The bursting pressure of PTFE and autogenous vein grafts after laser application was determined to be more than 300 mm Hg. The breaking strength of 6-0 PTFE sutures was compared before and after receiving 200 joules by contact laser application. No significant difference was found; in a before laser application the breaking strength was 585.0 +/- 9.6 gm compared with 560.0 +/- 16.4 gm after laser application (n = 4, p greater than 0.1). However, 6 0 polypropylene sutures were easily melted after receiving 10 joules of laser energy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965258 TI - Finding the high-risk patient with coronary artery disease. PMID- 2965259 TI - Two superb vaccines against hepatitis B in Mexican standoff. PMID- 2965260 TI - [Suppressor T-cells generated in mice late after thermal injury]. PMID- 2965261 TI - Determination of the hypertrophic site of the left ventricle by body surface mapping in patients with hypertension. AB - To estimate the hypertrophic sites of the left ventricle by body surface mapping (MAP), we performed MAP in 55 patients with hypertension and compared the MAP data with echocardiographic findings. MAP data were analyzed using the departure map technique reported by Flowers et al. The mean and standard deviation (SD) of the normal control were obtained from 40 normal volunteers. We constructed departure maps at 20, 30, 40, 50 and 60 msec from the onset of the QRS. Each map indicates the area of abnormally increased potential outside the normal range at the time. Subjects were classified into 5 groups according to the appearance time of the abnormal positive area. Septal thickness was significantly increased in groups that had an abnormal positive area at 20 msec, and left ventricular posterior wall thickness was significantly increased in the groups that had an abnormal positive area at 60 msec. We postulate that the increased electrical potential due to hypertrophy of the interventricular septum is represented by the abnormal positive area at 20 msec, and the increased potential of the left ventricular posterior wall by the abnormal positive area at 60 msec. MAP, especially the departure map technique, is a useful method to detect the abnormal electrical potential distribution in patients with left ventricular hypertrophy. PMID- 2965262 TI - [Atrial natriuretic peptides. Atrial natriuretic peptide precursors and gene structures]. PMID- 2965263 TI - [Atrial natriuretic peptide. Factors regulating release of atrial natriuretic peptide and its metabolism]. PMID- 2965264 TI - [Atrial natriuretic peptides. The function of the atrial natriuretic peptide as hormone and neuropeptide]. PMID- 2965265 TI - [Atrial natriuretic peptides. Radioimmunoassay]. PMID- 2965266 TI - [Atrial natriuretic peptides. b. Atrial natriuretic peptide in edematous disorders]. PMID- 2965267 TI - [Atrial natriuretic peptides. c. Endocrine diseases]. PMID- 2965268 TI - [Evaluation of the assay for alpha 2PI and alpha 2PI-plasmin complex using the one-step sandwich method (EIA)]. PMID- 2965269 TI - [Measurement and clinical significance of protein S by immunological assay]. PMID- 2965270 TI - [The identification of inhibitors of acantholysis of pemphigus vulgaris]. PMID- 2965271 TI - Central dipsogenic effect of synthetic rat atrial natriuretic polypeptide in normotensive rats. AB - The effect of intracerebroventricular (i.c.v.) injection of synthetic rat atrial natriuretic polypeptide (alpha-rANP) on drinking behavior was studied in normotensive rats, alpha-rANP (0.2, 0.4 or 0.8 micrograms in 5 microliter) caused a dose-dependent dipsogenic effect which was abated by i.c.v. pretreatment with saralasine (9 micrograms in 5 microliter). These results suggest that alpha-rANP possesses dipsogenic effects in water repleted rats and that brain angiotensin is involved. In addition, our data indicate that, at least as far as the effect of cerebral ANP is concerned, there are some differences between alpha-rANP and human atrial natriuretic polypeptide. PMID- 2965272 TI - [Origin of the pain syndrome in asymmetric myocardial hypertrophy in patients with stable hypertension]. AB - A study of 20 patients with essential hypertension, stage IIB, asymmetrical myocardial hypertrophy and chest pains has suggested that the pain syndrome, presenting as "possible angina", positive functional tests and reduced label accumulation around the ventricular septum may be indicative of coronary insufficiency. PMID- 2965274 TI - [Endoscopic methods of the treatment of acute biliary pancreatitis]. PMID- 2965273 TI - A thromboxane synthetase antagonist ameliorates progressive renal disease of Dahl S rats. AB - Recent studies have suggested a role for thromboxane in the progression of renal disease. The current study evaluated the role of this arachidonic acid metabolite in a model of renal disease which bears many biologic similarities to that in the kidneys of patients with chronic progressive renal failure. The model is that induced by ferritin-anti-ferritin immune complex nephritis in Dahl-salt sensitive rats rendered hypertensive by a high salt intake. Rats with this model of renal disease were chronically given a thromboxane synthetase antagonist OKY-046 or a placebo treatment from 16 to 29 weeks of age. Sequential observations of serum creatinine and 24-hour urinary protein excretion showed an ameliorating effect of OKY-046 on these renal parameters. Histologic examination of the kidneys also showed significantly less glomerular sclerosis in OKY-046 treated animals. The efficacy of OKY-046 was monitored by measurements of serum TXB2 levels and of glomerular production of TXB2 (and other prostaglandins); amounts of TXB2 were significantly reduced in the OKY-046 group. It is concluded that blockade of thromboxane generation has been successful in ameliorating the functional and structural lesions in this model of renal disease, providing further support to the thesis that thromboxane is an important mediator in events leading to eventual chronic renal failure and sclerosis. PMID- 2965275 TI - [Laparoscopic diagnosis of chronic calculous and acalculous cholecystitis]. PMID- 2965276 TI - [Clinico-anatomical aspects of treating coccygodynia]. PMID- 2965277 TI - [Acute complicated cholecystitis in middle-aged and elderly subjects]. PMID- 2965278 TI - [Treatment of pancreatogenic peritonitis]. PMID- 2965279 TI - [A prominent man in Zemstvo medicine, D.S. Shchetkin]. PMID- 2965280 TI - Aseptic necrosis of bone in ICR mice. AB - Aseptic bone necrosis was observed in the tibia of 23 ICR mice. Histological changes were characterized by a loss of marrow tissue with proliferation of connective tissue and bone necrosis with empty osteocytic lacunae. Focal necrosis was confined beneath the articular cartilage. Extensive necrosis was present in half or all of the epiphysis. Massive necrosis was noted in the diaphysis of one animal. It was considered that focal necrosis might be related to degenerative osteoarthritis, and that extensive and massive necroses might have been caused by a disturbance of the blood supply. PMID- 2965281 TI - The initiation and maintenance of a colony of coccidia-free wood mice (Apodemus sylvaticus). AB - Wood mice from a population showing enzootic infection with Eimeria were trapped and bred under laboratory conditions. The maintenance regime, which freed the animals from coccidia, and the breeding procedures used over several generations are described and discussed. PMID- 2965282 TI - Turn taking in atypical conversations: the case of the speaker/augmented communicator dyad. AB - Structural disturbances in conversation and their consequence for the distribution of control between partners were illustrated in a study investigating turn taking in dyads involving speech-handicapped individuals using alternative communication system (augmented or aided communicators) and normally speaking adults (unaided communicators or speakers). The turn-taking system in these atypical conversations differed from normal conversations in several ways. Interactants had to adapt to specific physical constraints on the interaction and compensated for these constraints by using "available" behaviors to signal turn exchanges. Augmented communicators were unsuccessful in interaction management as evidenced in the unaided communicators' unilateral control over them in turn regulation. Augmented communicators were frequently unsuccessful in their attempts to secure speaking turns, whereas unaided communicators were extraordinarily successful as compared with typical conversation. Implications for the understanding of normal and atypical interaction patterns and their relevance to conversational control are discussed. PMID- 2965283 TI - A new coronary tube for Cabrol's technique. PMID- 2965284 TI - Electrical stimulation of hypothalamic arcuate nucleus (ARC) can change responses to electroacupuncture of neurons of dorsal raphe nucleus (DR) and locus coeruleus (LC). PMID- 2965285 TI - Effect of laser-heated tip angioplasty on human atherosclerotic coronary arteries. AB - Effects of laser-heated-tip angioplasty on arteries is not fully understood. We report histology, ultrasound image properties, and vasoreactivity of human atherosclerotic coronary arteries after exposure to the laser-heated tip catheter. Segments of isolated human coronary arteries were obtained within 5-8 hours postmortem. Side branches were ligated and perfused with Krebs-Ringer solution. Coronary occlusions were recanalized during perfusion using a 1.5-mm tip heated twice with a 10 W argon laser for 10 seconds while two-dimensional 12 MHz ultrasound images were recorded. Images documented vessel recanalization and an increase in ultrasound refractile properties of vessel walls adjacent and 2-5 mm distal to the heated tip. Histologic studies showed charring along the neolumen and extensive coagulation pattern within the plaque. Vasoreactivity was assessed by measuring flow rate changes during perfusion with 100 ml of 10(-5) M serotonin followed by washout with serotonin-free solution. Recanalized arteries showed a 50% increase in magnitude of vasospasm, which was persistent for 5 hours, compared to control atherosclerotic vessels, which relaxed within 30 minutes. In conclusion, laser-heated-tip-irradiated vessels demonstrated plaque coagulation, increased ultrasound refractile properties of plaque, and increased vasospasm, which persisted for several hours. PMID- 2965286 TI - Laser balloon angioplasty: effect of tissue temperature on weld strength of human postmortem intima-media separations. AB - Dehiscence of portions of atheromatous plaques fractured during percutaneous transluminal coronary angioplasty may contribute to both abrupt reclosure and gradual restenosis. Laser balloon angioplasty has been shown to be effective in welding human plaque-arterial wall separations in vitro by heating tissues with a Nd:YAG laser during balloon inflation. To define the potentially useful therapeutic range of tissue temperature required to achieve thermal welds, 220 1 cm diameter discs of human postmortem atheromatous aortic tissue, the intimal plaque of which had been separated from the media, were exposed to 3-25 watts of Nd:YAG laser radiation delivered over a 12-mm2 nominal spot size for 20 seconds via a 400-micron core optical fiber. As measured with a thermistor, adventitial temperature reflected the temperature at the plaque-media junction to within 10 degrees C. The degree of tissue temperature elevation was related to delivered energy, while effective tissue penetration increased to maximum depth of 3 mm at the highest power density. Strength of tissue welds was defined as the force required to shear opposing layers of welded segments. Adventitial tissue temperatures below 80 degrees C were not associated with appreciable welds, while equilibrium temperatures between 95 degrees C and 140 degrees C were consistently associated with effective mean weld strengths, which increased linearly from 25 to 110 g, respectively. Temperatures greater than 150 degrees C were associated with rapid tissue dehydration and charring. These data suggest that the therapeutic range of tissue temperature that provides effective thermal fusion of intima-media separations is broad and that the depth and degree of thermal coagulation can be controlled by manipulation of laser energy delivery. PMID- 2965287 TI - Laser balloon angioplasty: effect of constant temperature versus constant power on tissue weld strength. AB - Thermal fusion of intimal plaque with the arterial wall during coronary balloon angioplasty may significantly reduce the incidence of abrupt closure and may reduce the occurrence of delayed restenosis by improvement of luminal size and shape. Although Nd:YAG laser energy has been shown to be effective in the thermal fusion of plaque-arterial wall separations in vitro, the most efficient manner of energy delivery for rapid achievement of therapeutically useful equilibrium tissue temperature during laser exposure has not been defined. A comparison of weld strength achieved was therefore made between two formats of laser delivery: constant power vs. decremental power with an initial high dose followed by the minimal serial decrements necessary to maintain tissue temperature constant for 15 seconds. One hundred sixty-six tissue discs of human postmortem aorta of 11 mm diameter were studied. Intimal plaque was separated from the media, the two layers were juxtaposed, a force of 4 pounds was applied, and a fiberoptic delivered laser beam was directed perpendicular to the tissue over a 3-mm diameter nominal spot size. Weld strength was measured as the shear force required to separate completely the two tissue layers. The mean weld strength (75 g) achieved by use of the decremental power format was significantly higher (P less than .01) than the mean strength (32 g and 56 g) achieved by using constant power for 20 and 30 seconds, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965288 TI - Pulsed laser angioplasty: wavelength power and energy dependencies relevant to clinical application. AB - Despite the theoretical advantages of submicrosecond pulsing for clinical laser angioplasty systems, the optimal laser parameters for clinical application are undefined. Further, the enormous peak powers achieved by submicrosecond pulses destroy available fiberoptics. We irradiated 797 segments of cadaver atherosclerotic aorta with nanosecond pulses at 266, 308, 355, 532, and 1064 nanometers. Effective cutting occurred at lower energy fluences in the ultraviolet than in the visible or infrared. For 308 nanometers, at any energy density, number of pulses to perforation was relatively independent of power density. Therefore, long-pulse ultraviolet wavelengths which could be transmitted through fiberoptics were identified as well suited for a clinical, submicrosecond pulsed laser angioplasty system. PMID- 2965290 TI - Gas volume quantitation during argon ion laser ablation of atheromatous aorta in blood and 0.9% saline media with an optically shielded catheter. AB - Using an optically shielded fiber optic laser catheter, the amount of gas produced when firing an argon ion laser into 0.9% saline solution or blood alone and into atheromatous aorta in either a blood or 0.9% saline medium was quantitated. Energies from 0.25 to 4 joules (J) were used at powers of 2, 5, and 8 W. We found that total volume of gas produced is small not only at equilibrium (0.3 +/- 0.1 microliter/J when firing in blood alone and also when ablating aorta in blood or saline media) but also at peak (2.5 +/- 0.2 microliters/J firing in blood alone and 1.0 +/- 0.1 microliter/J or 0.9 +/- 0.1 microliter/J when ablating aorta in saline or blood, respectively). Because these volumes are small, a clinically significant event from a gas embolus is unlikely during intravascular laser ablation of atheromatous plaque in the energy and power range studied. No gas was quantitated when firing the argon ion laser into 0.9% saline solution alone. The peak gas volume when firing in blood alone was significantly greater than that produced in the other chamber environments. This is thought to be due to increased absorption of argon laser light by hemoglobin. The gas volumes produced by lasing aorta in 0.9% saline or blood were not statistically different. PMID- 2965289 TI - Effects of varying argon ion laser intensity and exposure time on the ablation of atherosclerotic plaque. AB - Using continuous wave (CW) argon ion laser light, a total of 253 laser exposures of varying power (1.5, 3, 5, 8 or 10 W) and duration (20-1,333 ms) were delivered to four segments of human atheromatous aorta obtained at autopsy. Exposure conditions were controlled by using an optically shielded laser catheter that provided a 500 micron spot of light of known power. Two thresholds for consistently reproducible ablation could be defined-an intensity threshold at 25.5 W/mm2 and a fluence threshold at 3.2 J/mm2. Above threshold, a fluence of 5.1 J/mm2 was found to produce the most efficient ablation, ie, removed the greatest volume (mm3) per energy delivered (J) compared to other fluence levels employed (p less than 0.0001). Between aortic segments, however, considerable variability in efficiency (mm3/J) was observed, possibly owing to different optical properties and/or plaque composition. Low-intensity laser radiation produced inconsistent ablation and extensive coagulation effects to surrounding tissue. When a fluence of 5.1 J/mm2 was constructed with a high-intensity laser beam and a short exposure time, consistent and efficient tissue removal resulted without histologic evidence of coagulation necrosis. PMID- 2965291 TI - Nd-YAG laser energy distribution in an artificial obstruction: influence of lasing parameters in a model of laser angioplasty. AB - A Nd-YAG laser (1064 nm) coupled to a silica fiber (0.6 mm core diameter) was used to create defects in a model of arterial vascular obstruction. We employed transparent agar doped with black ink as atheromatous material and studied the size and shape of defects created by various lasing parameter settings. By adding calcium sulphate to the agar its scattering properties were enhanced. The created defects correspond to a temperature boundary. The optical properties of the agar greatly influenced the size and shape of the created defects. In the agar with enhanced scattering properties, the created defects showed an unfavourable penetration width-depth ratio. Maximum width of penetration always exceeded the fiber diameter. This may contribute to an increased risk of vessel wall perforation in small vessels and, if the fiber is positioned close to the vessel wall, even when a coaxial position is maintained. With increasing cumulative energy, both maximum depth and width of penetration leveled off in both agars. The results suggest that agar can be used to obtain empirically the lasing parameters that will minimize the risk of vessel wall perforation by an axially positioned fiber. The agar model needs further study to determine its limitations, but agar seems to be a useful substitute for atheroma in the study of laser catheter angioplasty. PMID- 2965292 TI - Cardiovascular effects of intravenous and intracoronary administration of atrial natriuretic peptide in halothane anesthetized dogs. AB - Cardiovascular actions of synthetic 1-28 human natriuretic peptides (hANP) were examined in dogs anesthetized with halothane. In seven closed-chest dogs a Swan Ganz catheter was inserted for measurement of cardiac output. Intravenous infusion of increasing doses of hANP (0.1, 0.3, 0.9 microgram/kg/min) lowered mean aortic pressure without affecting heart rate significantly. Cardiac output and pulmonary wedge pressure were markedly decreased while total peripheral resistance was increased significantly. All these parameters returned to control levels after 1 hr of recovery with an 100-150ml of saline infusion to increase pulmonary capillary wedge pressure to the preinfusion value. Intracoronary infusion of hANP (0.05 and 0.1 microgram/kg/min) did not cause any significant changes in coronary flow and regional contraction. These results indicate that the hypotensive action of hANP is due to a decrease in cardiac output mediated by reduced preload but not by negative inotropic action. PMID- 2965294 TI - Analgesia and plasma beta-endorphin-like immunoactivity in compound 48/80-induced hypovolemia of the rats. AB - The effects of subcutaneous (s.c.) administration of compound 48/80 (a well known histamine liberator) on latency to thermoalgesic stimulus, hematocrit (Hct) and plasma levels of beta-endorphin-like immunoreactivity (beta-END-LI) were investigated in male rats. The s.c. administration of compound 48/80 in doses ranging from 0.5 to 5.0 mg/kg into the rats produced significant analgesia in the hot plate test and increased Hct in a dose-dependent manner. Concomitant variation was observed between the analgesia and the increase of Hct. This analgesic effect, but not the increase of Hct, was diminished by pretreatment with the opiate receptor antagonist, naloxone (5 mg/kg, s.c.). A significant increase of plasma beta-END-LI was observed by s.c. injection of compound 48/80. Together with a previous finding that compound 48/80 induced-hypovolemia increases the renin release from kidney and then causes water intake in the rats, it is suggested that s.c. administration of compound 48/80 induced analgesia mediated through stimulation of an opioid system, may be closely related to stimulation of the renin-angiotensin system. PMID- 2965293 TI - Ganglioside treatment of diabetic rats; effects on nerve adenosine triphosphatase activity and motor nerve conduction velocity. AB - Ouabain-sensitive ATPase activity (expressed as nmol ADP produced/h/mg (wet) nerve +/- SEM) was measured in homogenates of sciatic nerve from control rats and rats with streptozotocin-induced diabetes of 8 wk duration. Nerves from diabetic rats showed activity (21.7 +/- 2.0) which was significantly (p less than 0.05) less than that of controls (34.6 +/- 4.8). These animals also showed a deficit in conduction velocity (m/sec +/- SEM) of sciatic nerve motoneurones (50.7 +/- 0.4 vs. 57.7 +/- 0.7 in controls; p less than 0.001). In parallel, matched control and diabetic groups were treated daily with mixed gangliosides extracted from bovine brain (10 mg/kg i.p.). After such treatment for 8 wk the deficit in ouabain-sensitive ATPase activity did not develop in the diabetic group (treated diabetics, 31.9 +/- 3.7; treated controls, 34.5 +/- 3.8). However, the treatment did not affect the deficit in motor nerve conduction velocity (treated diabetics, 50.9 +/- 1.1 vs. treated controls, 57.9 +/- 0.5; p less than 0.001). Accumulations of the polyol pathway metabolites--sorbitol and fructose--together with depletion of nerve myo-inositol were similar in both diabetic groups. These data indicate an etiology for the conduction velocity deficit which differs from that of the deficit in ouabain-sensitive ATPase. PMID- 2965296 TI - Hospitalizations due to tap water scalds, 1978-1985. PMID- 2965295 TI - Liposomes in chemo- and immunotherapy of cancer. AB - In this paper, we report on the in vivo behavior of liposomes as a function of their size and composition. It is emphasized that by varying these parameters we can influence not only the rate of blood elimination but also the intrahepatic destination of the liposomes. Thus, we show that small liposomes with diameters well below 100 nm can reach and be internalized by the parenchymal cells of the liver, i.e. the hepatocytes. The rate and the extent at which this occurs depends on the liposomal composition. With respect to the application of liposomes as a drug carrier system in anticancer therapy, we put emphasis on the liver macrophage, i.e. the Kupffer cell, as a target cell. Large liposomes with diameters well over 100 nm exclusively are taken up by these cells as far as hepatic uptake is concerned. By encapsulation within liposomes, a drug may be delivered specifically to these macrophages; this will prevent its rapid excretion from the body and/or undesired accumulation in other cell types. Two examples of the way in which this condition may be exploited are presented. First, we demonstrate the formation of intracellular depots in the macrophages of the cytostatic drug 5-fluorodeoxyuridine (FUdR), thus preventing the rapid metabolism of the drug by the hepatocytes and allowing its sustained release from the macrophages and subsequent uptake by adjacent metastatic tumor cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965297 TI - Disparate effects of 5-bromodeoxyuridine on sister-chromatid exchanges and chromosomal aberrations in Bloom syndrome fibroblasts. AB - The existence of a high frequency of spontaneous sister-chromatid exchanges (SCEs) in Bloom syndrome (BS) has thus far been supported by data on a small number of BS cell lines. To examine the cause of baseline SCEs more broadly, the frequencies of SCEs, as well as chromosomal aberrations (CAs) in 4 additional BS fibroblast strains were compared, under different assay and cell culture conditions, with those of normal cells in the range of approximately 0.9-90% 5 bromodeoxyuridine (BrdUrd) substitution into template DNA. SCEs at low levels of BrdUrd substitution were detected by an extremely sensitive immunofluorescent technique. From approximately 0.9% to 4.5% BrdUrd substitution, the SCE frequency in BS cells remained constant, at a level (40/cell) 8 times higher than that of normal cells. As BrdUrd substitution increased further, the SCE frequency in BS cells increased almost linearly, reaching 70-100 per cell at approximately 90% substitution, while the SCE increment in control fibroblasts was less than 5 per cell. Analysis of SCEs in 3 successive replication cycles similarly revealed that the SCE increment in BS cells depended on BrdUrd only at a high BrdUrd substitution level. In contrast to data on SCEs, CA induction by incorporated BrdUrd in BS cells was only slightly higher than that in normal cells. Thus, BS cells are extremely sensitive to BrdUrd for SCE induction, but much less so for CA induction. PMID- 2965298 TI - Gene expression in E. coli after treatment with streptozotocin. AB - Gene induction by the methylating agents streptozotocin (STZ), N-methyl-N nitrosourea (MNU), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was evaluated in E. coli fusion mutants. These mutants have fusions of the lac operon to genes induced by treatment with sublethal levels of alkylating agents and were previously selected from random insertions of the Mu-dl (Apr lac) phage by screening for induction of beta-galactosidase activity in the presence of methyl methanesulfonate or MNNG. The results demonstrate that STZ differs from MNNG and MNU in failing to induce aidC expression. Further, expression of aidC after exposure to MNU and MNNG occurs only in nonaerated cultures; aeration blocks the induction. Induction of aidD, alkA, aidB, and sfiA expression occurs with all 3 agents although at markedly lower concentrations of MNNG and STZ compared to MNU. alkA and to a lesser extent aidD mutants of E. coli strains were more sensitive to these agents, while no differences were evident between wild-type and aidB or aidC fusion mutants. PMID- 2965299 TI - Induction of chromosome damage and sister-chromatid exchanges in human lymphocyte cultures by the antitumour antibiotic Neocarzinostatin. AB - Neocarzinostatin (NCS), a chemotherapeutic antibiotic, was investigated for the ability to induce chromosomal aberrations and sister-chromatid exchanges (SCEs) in human lymphocyte cultures. It was observed that the antibiotic causes a cell cycle delay and reduces the mitotic index. Analysis of the induced chromosomal abnormalities showed that they are mainly chromosome and chromatid breaks; while the frequency of SCEs was increased, the magnitude indicates that NCS cannot be considered a potent inducer of SCEs. PMID- 2965300 TI - Hexosaminidase A activity and amyotrophic lateral sclerosis. AB - Abnormalities of GM2 ganglioside metabolism owing to hexosaminidase A (Hex A) deficiency have been associated with ALS phenotypes. The clinical features described in these ALS patients with Hex A deficiency include early onset, positive family history, and/or long disease duration. In an attempt to determine prospectively the incidence of Hex A deficiency within an ALS population, the records of The Mount Sinai Medical Center ALS Clinic were reviewed to select those patients with "atypical" ALS (total N = 52), i.e. onset before age 35, positive family history, and/or disease duration greater than 90 months. The control group (total N = 50), "typical" ALS patients, did not fulfill any of these historical criteria. Hex A activity determined in isolated peripheral blood leukocytes was normal in all typical ALS patients (mean 67.3%). Hex A deficiency was not found in any atypical ALS patients. Thus, Hex A deficiency apparently is an unusual etiology of typical or atypical ALS but is of medical and genetic importance in individual families. PMID- 2965301 TI - Hemodynamic and coagulation responses to 1-desamino[8-D-arginine] vasopressin in patients with congenital nephrogenic diabetes insipidus. AB - The antidiuretic hormone arginine vasopressin interacts with two types of receptors: V1, which mediates the effects of vasopressin on vascular smooth muscle, and V2, which mediates the antidiuretic effects on renal tubules. Resistance of the renal tubules to arginine vasopressin and to the antidiuretic V2-specific agonist 1-desamino[8-D-arginine] vasopressin (dDAVP) occurs in congenital nephrogenic diabetes insipidus, a rare X-linked disease, although the V1-receptor responses remain intact. The extrarenal actions of dDAVP in normal persons are a decrease in blood pressure, an increase in plasma renin activity, and stimulation of the release of factor VIIIc and von Willebrand factor. We measured the response of mean arterial pressure, pulse rate, plasma renin activity, factor VIIIc, and von Willebrand factor to an infusion of dDAVP (0.3 microgram per kilogram of body weight) in seven male patients with congenital nephrogenic diabetes insipidus, six obligatory carriers of the gene for nephrogenic diabetes insipidus, five patients with central diabetes insipidus, and four normal subjects. In the normal subjects and the patients with central diabetes insipidus, dDAVP decreased mean arterial pressure (by 10 to 15 percent) and increased pulse rate (by 20 to 25 percent), renin activity (by 65 percent), and the release of coagulation factors (twofold to threefold) (all changes were significant, P less than 0.01). None of these changes were observed in the patients with congenital nephrogenic diabetes insipidus, and minimal responses were observed in the obligatory carriers. These results confirm the existence of extrarenal vasopressin V2-like receptors, which may be defective in patients with congenital nephrogenic diabetes insipidus. PMID- 2965303 TI - Teicoplanin for patients allergic to vancomycin. PMID- 2965302 TI - A prospective study of platelets and plasma proteolytic systems during the early stages of Rocky Mountain spotted fever. AB - We prospectively examined early changes in platelets and plasma proteolytic systems in 12 vaccinated and 6 unvaccinated volunteers in whom Rocky Mountain spotted fever developed after challenge with Rickettsia rickettsii. The platelet counts declined while the plasma concentration of beta-thromboglobulin and the ratio of beta-thromboglobulin to platelet factor 4 increased, indicating in vivo activation of platelets. Plasma levels of antithrombin III decreased and levels of fibrinopeptide A increased, indicating in vivo activation of the coagulation system. Plasma fibrinogen levels peaked at 24 hours and gradually declined; this is consistent with the behavior of fibrinogen as an acute-phase reactant. Prolongation of the prothrombin time and a decrease in plasma levels of factor VII in the absence of evidence of liver injury suggested possible activation of the extrinsic pathway of coagulation. A decline in plasma prekallikrein levels with an increase in plasma C1-inhibitor-kallikrein complexes suggested activation of kallikrein, probably through the intrinsic coagulation system. Elevations in levels of plasma fibrin-degradation products and alpha 2-antiplasmin-plasmin complexes with declines in plasminogen and alpha 2-antiplasmin levels provided evidence of activation of the fibrinolytic system. Elevated plasma levels of tissue plasminogen activator and von Willebrand factor reflected endothelial stimulation. Thus, even early in the course of Rocky Mountain spotted fever that is treated promptly, there is activation of platelets, coagulation pathways, and the fibrinolytic system. These changes may be related to endothelial perturbation, a major pathogenetic mechanism in the disorder. PMID- 2965304 TI - South Africa's first genetically engineered vaccine. PMID- 2965306 TI - Receptor-directed focusing of lymphokine release by helper T cells. AB - The interaction between helper T cells and B cells, leading to the production of antibody to thymus-dependent antigens, was the first cell interaction clearly defined in the immune system; it remains both paradigmatic and controversial. Two requirements of this interaction, that the helper cell (TH) and the B cell must recognize antigenic determinants that are physically linked, and that the TH and the B cell must share genes encoding major histocompatibility complex (MHC) class II molecules, led to the concept that TH-B interaction required an intimate physical association of the two cell types. But in vitro studies have shown that TH can be replaced by soluble, antigen-nonspecific factors, capable of activating any B cell to secrete antibody. We have previously proposed that the requirements for TH-B contact might result from TH cells releasing their lymphokines in a polar fashion directed at that portion of the cell membrane where T-cell receptor cross-linking is actually occurring. Using an artificial monolayer of a cloned helper T-cell line, we show that lymphokines are released preferentially over the area of receptor cross-linking under conditions of limited TH-cell activation. Thus, it appears that one important aspect of the specificity of TH-B cell interactions is the receptor-directed polar release of helper lymphokines. PMID- 2965305 TI - Heparan sulphate bound growth factors: a mechanism for stromal cell mediated haemopoiesis. AB - The proliferation and development of haemopoietic stem cells takes place in close association with marrow stromal cells. This intimate cell contact presumably enables the stem cells and their progeny to respond to stimuli present on the stromal cell surface. While the nature of these stimuli has not been determined, it is likely that growth factors play some role. Recently, it was demonstrated that the natural and the recombinant haemopoietic growth factor, granulocyte/macrophage colony stimulating factor (GM-CSF), could be adsorbed out of solution by an extract of human marrow stromal extracellular matrix (ECM) with retention of biological activity. However, the precise ECM molecules involved were not identified. Here, we clearly demonstrate that the major sulphated glycosaminoglycan of mouse marrow stroma, heparan sulphate, possesses the ability to adsorb both GM-CSF and the multilineage haemopoietic growth factor, Interleukin 3 (IL-3). Furthermore, these growth factors, once bound, can be presented in the biologically active form to haemopoietic cells. PMID- 2965309 TI - Treatment of acute cardiac tragedies. By A. C. Morgan, 1927. PMID- 2965308 TI - Inhibition of 5-hydroxytryptamine accumulation and deamination by substituted phenylalkylamines in hypothalamic synaptosomes from normal and reserpine pretreated rats. AB - 1. In the present study the abilities of different compounds to inhibit MAO inside and outside the serotonergic neurons, to inhibit the accumulation of 5-HT and to release 5-HT were separated by using different in vitro techniques. With these methods a number of substituted phenylalkylamines, which are reversible inhibitors of monoamine oxidase (MAO) type A, were characterized. 2. The compounds were examined regarding their ability to inhibit the accumulation of 5 HT and to inhibit MAO in the same synaptosomal preparation of hypothalamus from normal and reserpine-pretreated rats. The difference in the uptake of 14C-5-HT (0.1 mumol/l) in the absence and presence of citalopram (0.25 mumol/l) was taken as a measure of the accumulation into the serotonergic synaptosomes. The deamination of 14C-5-HT (0.1 mumol/l) in the presence of citalopram (0.25 mumol/l) was considered as that brought about outside the serotonergic synaptosomes, whereas the difference between the deamination in the absence and presence of citalopram was taken as the MAO activity inside the serotonergic synaptosomes. 3. Most of the phenylalkylamines were slightly more potent as MAO inhibitors outside serotonergic synaptosomes than as inhibitors of 5-HT accumulation in normal rats. The most potent MAO inhibitors, both in absolute terms and in comparison with uptake inhibitory potency, were the 2,6-dichloro (FLA 365) and the phenylpropylene-(FLA 417) derivatives. 4. A difference in potency on the accumulation in synaptosomes from normal and reserpine-pretreated rats was found for many of the phenylalkylamines with the exception of FLA 365, FLA 417 and the 2,5-dimethyl derivative RAN 113.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965307 TI - Inhibition of pentose cycle of A549 cells by 6-aminonicotinamide: consequences for aerobic and hypoxic radiation response and for radiosensitizer action. AB - Metabolism of glucose via the pentose cycle is a principal source of NADPH, an important cellular reducing species. Both aerobic and hypoxic irradiation stimulate the pentose cycle activity of A549 human lung carcinoma cells, which indicates that NADPH is utilized during irradiation, either as a direct hydrogen donor or as a cofactor for enzymatic repair of radiation damage. To evaluate the role of the pentose cycle in radiation response, we treated A549 cells with 6 aminonicotinamide (6-AN), which blocks the oxidative limb of this pathway in some cell lines. We found 6-AN to be a very effective inhibitor of pentose cycle activity, as indicated both by accumulation of 6-phosphogluconate in A549 cells and by the inability of nitrofurazone or peroxide to stimulate release of 14CO2 from 14C-1-labeled glucose after 6-AN treatment. Effects of 6-AN were time and concentration dependent; it caused partial inhibition of glycolysis but had no effect on respiratory rate or on intracellular glutathione levels. Effects of 6 AN on radiation response were examined under two conditions: 1) after treatment with 0.3 mM drug for 5 hours, which inhibited pentose cycle activity by 50%, and 2) after treatment for 15 hours, which completely inhibited pentose cycle activity. Neither treatment affected aerobic radiation response, but both increased hypoxic sensitivity to a similar extent, with the oxygen enhancement ratio reduced from 3.0 to 2.0 at a 0.05 surviving fraction. Treatment of A549 cells with 6-AN caused an increase in hypoxic cell radiosensitization by misonidazole, but effects of the combined agents were not more than additive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965310 TI - Mucopolysaccharides in gastric cancer. AB - The histochemical properties of mucus in 48 diffuse type gastric carcinomas and in 46 intestinal type gastric cancers were investigated. In the diffuse type there was a prevalence of intracellular secretion of mucopolysaccharides with a significantly larger portion of sulfated mucopolysaccharides. Acid mucopolysaccharides occurred in the tumor cells only along with neutral ones. In the intestinal type of this cancer, in turn, extracellular secretion prevailed, along with the occurrence of purely acid mucopolysaccharides and a smaller portion of sulfated ones. The secretion of pure neutral mucopolysaccharides in both types of gastric cancer was very low. In the marginal areas of intestinal type carcinomas, incomplete intestinal type IIb metaplasia was frequently found. PMID- 2965311 TI - Prostacyclin (epoprostenol) as the sole antithrombotic agent in postdilutional hemofiltration. AB - Prostacyclin (PGI2), the most potent and short-lived antiplatelet agent known today, has been used successfully as an antithrombotic in hemodialysis (HD). However, its vasodilatory effect has been the source of blood pressure instability in acetate HD and has restricted its use to bicarbonate HD. The authors took advantage of the better cardiovascular stability obtained with hemofiltration (HF) to compare the effects of PGI2 versus heparin either with acetate or bicarbonate HF in 4 patients. Efficacy of PGI2 in preventing thrombosis of the extracorporeal circuit was demonstrated in all cases with a dose of 4 ng/kg/min. HF performances remained unaffected whatever antithrombotic agent was used. Platelet activation as shown by BTG and PF4 release was inhibited by the PGI2 infusion. Platelet proteins release was greater with acetate HF, suggesting that acetate may have a specific role in platelet activation. Although the use of PGI2 was straightforward, it is worth noting that PGI2 partially suppressed the cardiostability usually associated with acetate HF. We conclude that the efficacy of PGI2 was well maintained in spite of conditions of high platelet shear stress conditions, but also that PGI2 potentiated the vasodilatory effect of acetate and suppressed partially the cardiovascular benefits of HF. PMID- 2965312 TI - [Clinical symptoms and the results of Doppler ultrasonography in stenosis and occlusion of the internal carotid artery]. AB - In 28 patients with stenosis or occlusion of the external carotid artery confirmed by angiography the clinical manifestations were analysed, and the usefulness of Doppler ultrasonography of the arteries for diagnosis was assessed. The clinical manifestations of stenosis or occlusion of the external carotid artery include trophic and vascular signs. The most frequent trophic signs were: premature rapid loss of teeth and atrophy of facial muscles. The vascular clinical signs included weak pulsation of facial arteries. In 26 cases (96%) abnormal USG findings were obtained. The USG investigations showed that despite complete occlusion of the external carotid artery the blood flow in its branches was frequently maintained owing to collateral circulation. PMID- 2965313 TI - Corticosteroid effects on morphine-induced antinociception as a function of two types of corticosteroid receptors in brain. AB - The antinociceptive effect of parenterally and intracerebroventricularly injected morphine and beta-endorphin in adrenalectomized rats and in adrenalectomized rats treated with adrenal steroids was examined employing the hot-plate method. (1) Adrenalectomy sensitized the rats to an analgesic effect of morphine and beta endorphin. (2) Replacement therapy (chronic and acute) with corticosterone, dexamethasone or RU 28362 (glucocorticoid receptor agonist) effectively reversed the increase in the sensitivity to the analgesic effect of peripherally injected morphine (5 mg/kg i.p.) induced by adrenalectomy to the level of sham-operated animals. Glucocorticosteroids administered to non-adrenalectomized rats did not change the sensitivity to morphine. (3) Corticosterone had a biphasic, dose dependent effect; the most significant attenuation of the hypersensitivity to morphine-induced antinociception in adrenalectomized rats was achieved after 0.01 mg and after 10 mg (per kg b.w.). Doses of corticosterone of 0.005 mg/kg and in a range of 0.05-0.30 mg/kg were ineffective. (4) Corticosterone in a dose of 0.01 mg/kg (s.c.) had suppressant effects on the adrenalectomy-induced increase in the sensitivity to antinociception induced by morphine when given prior to morphine (60, 30 and 5 min) as well as after the injection of morphine (before the first and the second testing on the hot-plate, 15 and 5 min, respectively). (5) Intracerebroventricularly (i.c.v.) injected morphine and beta-endorphin also displayed the hypersensitivity to the analgesic effect in adrenalectomized rats which in both cases could be suppressed by 0.01 mg/kg of corticosterone given subcutaneously 5 min prior to administration of the opiate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965315 TI - Evidence for histaminergic arousal mechanisms in the hypothalamus of cat. AB - Polygraphic 23-hr recordings were carried out in 25 adult cats in order to examine the effects of both systemic and local injections of various histaminergic and antihistaminergic drugs on sleep-waking cycles. alpha Fluoromethylhistidine (alpha-FMH), a specific inhibitor of histidine decarboxylase, when injected intraperitoneally at a dose of 20 mg/kg, induced a significant increase in deep slow wave sleep (S2) and a decrease in wakefulness (W), without modifying light slow wave sleep (S1) and paradoxical sleep (PS). Intraperitoneal injections of mepyramine (1 mg and 5 mg/kg), a well-known histamine H1-receptor antagonist, increased deep slow wave sleep and decreased wakefulness, as well as paradoxical sleep. Bilateral injections of alpha-FMH (50 micrograms/1 microliter) into the ventrolateral posterior hypothalamus, where histamine immunoreactive neurones have been recently identified, resulted in a significant decrease in wakefulness and increase in deep slow wave sleep. Similarly, injections of mepyramine (120 micrograms/1 microliter) in the same structures caused a significant decrease in wakefulness and an increase in deep slow wave and paradoxical sleep as well. In contrast, local injections of SKF 91488 (50 micrograms/1 microliter), a specific inhibitor of histamine-N methyltransferase, led to a significant increase in wakefulness and decrease in both slow wave sleep (SWS) and paradoxical sleep. Injections of histamine, at doses of 5, 30 and 60 micrograms/1 microliter, also increased wakefulness and decreased slow wave sleep dose dependently, while these effects were completely blocked by pretreatment with mepyramine. The results suggest that histaminergic systems in the hypothalamus play an important role in arousal mechanisms and their actions are mediated through H1-receptors. PMID- 2965316 TI - The effect of selective D1 and D2 dopaminergic agents on sexual receptivity in the female rat. AB - A number of dopaminergic drugs was tested for their effect on female sexual receptivity in ovariectomised plus adrenalectomised rats primed with oestradiol benzoate. The selective D1 agonist (SKF 38393 5-40 mg/kg) and D1 antagonist (SCH 23390 0.1-10 mg/kg) had no significant effect on sexual behaviour. The results of administration of mixed dopaminergic agents (DOPA, 10-50 mg/kg with benserazide or 100-200 mg/kg alone and haloperidol 0.01-1.0 mg/kg) and selective D2 dopaminergic agents (LY 171555 2.5-800 micrograms/kg, BHT 920 0.01-1.0 mg/kg and sulpiride 5-80 mg/kg) indicate that a central dopaminergic system has an inhibitory control on female sexual activity exerted through D2 receptors. Any stimulatory effects of the exogenous agonists were probably due to an action on presynaptic D2 receptors. The predominance of the D2 pre- and post-synaptic receptor activity appears to be influenced by the sexual receptivity of the animal. PMID- 2965314 TI - Sensitivity of the response of 5-HT autoreceptors to drugs modifying synaptic availability of 5-HT. AB - The effect of midalcipran, an equipotent inhibitor of the uptake of 5 hydroxytryptamine (5-HT) and noradrenaline (NA), on the inhibition of release of 5-HT induced by lysergic acid diethylamide (LSD), was investigated by studying the overflow of transmitter from slices of hypothalamus from the rat prelabelled with [3H]5-HT. The (Z)-enantiomer of midalcipran, at 0.1 microM, displaced to the right the concentration-effect curve of inhibition of release of [3H]5-HT elicited by electrical stimulation induced by LSD. In contrast, the (E) enantiomer, which does not inhibit the uptake of monoamines, was devoid of any antagonizing effect. The inhibitory effect of 0.1 microM LSD was not modified in the presence of maprotiline, bupropion or mianserin at 1 microM. The results of this study show that the interaction between the 5-HT autoreceptor and the inhibitors of the uptake of 5-HT is related exclusively to their potency at inhibiting the uptake of 5-HT. The effect of the monoamine oxidase inhibitor, pargyline (1-10 microM), was also studied on the field-stimulated release of [3H]5-HT. The inhibitory effect of this drug was antagonized by methiothepin at 0.1 and 1 microM, by phentolamine 1 microM and 10 microM and by the inhibitor of the uptake of 5-HT, citalopram at 0.1 and 1 microM. The action of pargyline appeared to be mediated through the activation of the serotonergic autoreceptor and of the presynaptic alpha-adrenoceptor located on serotonergic nerve terminals. The results obtained with pargyline are consistent with the hypothesis of an interaction between the 5-HT autoreceptor and the uptake site for 5 HT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965317 TI - Infantile muscle glycogen storage disease: phosphoglucomutase deficiency with decreased muscle and serum carnitine levels. AB - We report a 5-month-old boy with recurrent vomiting, lethargy, and poor weight gain. He had profound metabolic acidosis and nonketotic dicarboxylic aciduria. The serum and muscle carnitine levels were significantly low (60% and 10% of the control means, respectively), suggesting that the patient had a systemic carnitine deficiency syndrome. The patient showed apparent clinical improvement on oral carnitine administration. A quadriceps muscle biopsy revealed a slight increase in intrafiber lipid droplets and mild accumulation of glycogen in the subsarcolemmal portion. An anaerobic glycolysis in vitro study showed a block after glucose-1-phosphate and before glucose-6-phosphate. Direct measurement of individual glycolytic enzymes in muscle of the patient demonstrated a marked decrease in phosphoglucomutase (PGM) activity (13% of the control mean). The specific defect of PGM activity in this patient suggests that the block in the anaerobic glycolytic pathway is the primary abnormality. PGM deficiency can be added as a newly recognized cause of secondary systemic carnitine deficiency syndromes. PMID- 2965318 TI - [Evaluation of the diastolic function of the left ventricle by Doppler echography in patients with acute myocardial infarction]. PMID- 2965319 TI - [Hypertrophy and left ventricular function following angiotensin-converting enzyme inhibition in hypertensive patients. Echocardiographic analysis]. PMID- 2965320 TI - [Doppler flowmetry in the perioperative study of the hemodynamics of supra-aortic trunks in neoplasms of the head and neck]. PMID- 2965321 TI - [Study of lymphocyte subpopulations in alcoholic liver cirrhosis]. PMID- 2965322 TI - A simple message. PMID- 2965323 TI - Feeding the handicapped child--a review of intervention strategies. AB - Feeding the handicapped child occurs three or four times per day, every day of the year, despite the child's feeding problems and, in the vast majority of cases, without life threatening consequences. Yet the need for adequate feeding therapy for these children is becoming increasingly recognised. This paper seeks to explore why intervention is so important. The complex causes of feeding problems will be reviewed briefly followed by a discussion of the reasons for intervention. Feeding therapy techniques, strategies and approaches will be reviewed with the need for parent involvement, early intervention and a 'feeding team' being stressed throughout. This paper highlights the need for intervention in feeding problems, whether or not they are life threatening, particularly in view of their effect upon quality of life of both the child and his family. PMID- 2965324 TI - Percutaneous transluminal angioplasty for the ischaemic lower limb: the early Dunedin experience. AB - A consecutive series of 60 percutaneous transluminal angioplasties has been performed in 53 patients with severe peripheral ischaemia over a 4 year period. There were 78 lesions dilated with an initial technical success rate of 78%. Forty-eight lesions were dilated for severe claudication and 30 for rest pain or necrosis. There was a mean followup of 24 months (range 9-56) for the immediately successful angioplasties. When these were analysed according to symptomatic relief, 59% of claudicants with iliac lesions were still improved at 2 years, compared with 86% of those with femoropopliteal lesions. Amongst patients treated for rest pain or necrosis there was a 100% success rate at 2 years for iliac lesions, compared with 49% for femoropopliteal lesions. The overall success of this procedure in this high risk group is reflected by the fact that at 2 years 85% of claudicants and 72% of those with rest pain and necrosis have avoided any further procedure. This encouraging early experience confirms the place of interventional radiology in the management of peripheral vascular disease. PMID- 2965325 TI - The American Academy of Pediatric Dentistry. Its scope and function. PMID- 2965328 TI - Androgen excess and the infertile woman. AB - Hyperandrogenism is not infrequently encountered in infertile women. However, elevated levels of androgen in blood may or may not be associated with hirsutism. Therefore, the presence of hirsutism alone may not be adequate to rule out hyperandrogenism as a factor in infertility. Although most patients with hyperandrogenism will have menstrual irregularity, some patients may give a normal menstrual history. Specific markers in blood may be used to direct therapy. Not all suppressive therapy is appropriate for use in the infertile patient. PMID- 2965327 TI - Gynecologic endoscopy in infertility. AB - Pelvic endoscopic techniques enable the physician to make accurate diagnoses, to treat specific pelvic disorders, and to gather information for research. This article focuses on these techniques as they apply to the management of infertile patients. Included for discussion are diagnostic and therapeutic laparoscopy and hysteroscopy, culdoscopy, and salpingoscopy. PMID- 2965326 TI - The association between chronic pelvic pain, psychiatric diagnoses, and childhood sexual abuse. AB - Twenty-five women with chronic pelvic pain who had undergone diagnostic laparoscopy and 30 women who had laparoscopic examinations for tubal sterilization or infertility investigation were compared psychologically using structured psychiatric and sexual abuse interviews. Results of the fiberoptic pelvic examination were rated independently using the American Fertility Society classification of endometriosis. Compared with controls, the patients with chronic pelvic pain showed significantly greater prevalence of lifetime major depression, current major depression, lifetime substance abuse, adult sexual dysfunction, and somatization. They were also significantly more likely than controls to have been a victim of childhood and adult sexual abuse. There were no significant differences in either the degree or type of pelvic disease between patients with pelvic pain and controls. PMID- 2965329 TI - Polymorphonuclear neutrophils function in untreated patients with chronic myeloid leukemia. AB - Morphologically mature polymorphonuclear neutrophils (PMN) isolated from the peripheral blood of 30 patients were examined. These cells manifested the diminution of phagocytosis, random migration, chemotaxis, thermotaxis and bactericidal capacity. The surface charge of circulating PMN and the percent representation of cells with receptor for Fc IgG were significantly decreased. Stimulated as well as unstimulated oxygen consumption, production of O2- and H2O2 were significantly decreased, whereas the hexose monophosphate shunt activity was similar to that noticed in controls. The aggregation of leukemic PMN was practically normal, however their adherence was significantly higher. The possible mechanisms responsible for the observed PMN defects are discussed. PMID- 2965330 TI - Broiler pulmonary hypertension syndrome. III. Commercial broiler strains differ in their susceptibility. AB - Broilers of 4 different commercial strains were kept during winter at an altitude of 1,350 m. The birds were slaughtered at weekly intervals and their relative right ventricular mass (pulmonary arterial pressure index = API values) determined. In addition, the incidence of ascites was recorded. Two of the strains suffered high losses from ascites, while the other 2 showed a certain degree of resistance. The former 2 groups had a higher mean API and a greater percentage of high API values in clinically normal birds than the other 2 groups. The group with the lowest incidence of ascites achieved the highest mean live mass at 51 days. There was also a decrease in mean API with age in clinically normal birds. There was no difference in the incidence of ascites between males and females. PMID- 2965331 TI - Bone trabecular pattern analysis in Down's syndrome with the use of computed panoramic tomography with a laser scan system. Quantitative analysis with the power spectrum method. AB - A quantitative analysis of the mandibular bone trabeculae of 44 patients with Down's syndrome and 68 normal persons by means of panoramic tomography was performed with a laser scan system. The results showed that the normal persons exhibited a correlation between the trabecular pattern and aging, whereas the patients with Down's syndrome showed no such correlation. These findings were consistent with geromorphism, which is one of the systemic features of Down's syndrome. PMID- 2965332 TI - [Atrial natriuretic peptides in liver cirrhosis associated with vascular decompensation]. PMID- 2965333 TI - [Opioid peptides and shock]. PMID- 2965334 TI - [Opioid peptides and neurohormonal reactions in stress and adaptation]. PMID- 2965336 TI - Human placental cells that regulate lymphocyte function. AB - We examined normal human placenta for an immunologic function by measuring the release of soluble inhibitory factor (SIF). SIF is a product of normal T lymphocytes and of the JEG-3 choriocarcinoma cell line, and blocks proliferative and antibody-producing responses of mononuclear cells. SIF can be further characterized by a noncovalently linked subcomponent, lipid suppressor substance. The villous surfaces of six normal human placentae were digested with collagenase to obtain a population of predominantly multinucleated giant cells. These cells were maintained in standard culture for 5 days after which the cell-free conditioned culture medium was assayed for SIF content by measuring suppression of [3H]thymidine incorporation into lymphocytes stimulated by low-dose phytohemagglutinin. Undiluted placental SIF induced 88% inhibition of this response (p less than 0.001). The placental SIF was found to contain lipid suppressor substance, as does SIF from mononuclear cells. We determined this by thin-layer chromatography where a peak of suppressive activity occurred at Rf 0.32 [( 3H]thymidine incorporation reduced from 21810 +/- 308 to 4121 +/- 214 cpm); this is the position on thin-layer chromatography to which mononuclear cell lipid suppressor substance migrates. Ion exchange chromatography comparing the elution patterns of lymphocyte-SIF and placental-SIF indicated that both eluted in the fraction of 40-50 mM PO4 means buffer, further suggesting identity between these two substances. SIF from placental and lymphocyte sources functioned by inducing the presence of suppressor cells in culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965337 TI - Modulation of T3 antigen in systemic lupus erythematosus (SLE). AB - One of the first steps in the T cell activation is modulation of T3-Ti complex from the cell membrane. This process was studied in patients with SLE by immunofluorescence staining and measured in a cell sorter. The modulation of T3 antigen was impaired in a number of patients. The mitogen-induced proliferation was also decreased in these cases, but decreased blast transformation occurred also with efficient modulation. Pretreatment of normal T lymphocyte with sera from patients with decreased modulation caused only moderate inhibition of T3 antigen modulation. Modulation in vitro was not influenced by steroids. PMID- 2965335 TI - [Use of immunocytochemical methods in the differential diagnosis of low differentiated cancer and lymphoma of the stomach (gastric biopsy specimens)]. PMID- 2965338 TI - Serum steroid hormone levels in systemic lupus erythematosus (SLE). AB - Dehydroepiandrosterone (D), D sulphate (DS), testosterone (T) and oestradiol (OE2) were determined by radioimmunoassay; cortisol (F) by a spectrofluorimetric method in the serum of 54 female patients with SLE. The values were evaluated in two age groups (32 patients with active ovarian function and 22 in menopause). The serum cortisol, T and DS levels were decreased, and there was no difference in that of free (unconjugated) D and OE2 concentrations compared to normal female subjects. PMID- 2965339 TI - Living together: people, animals, environment--a personal historical perspective. PMID- 2965340 TI - Studies on histaminergic compounds: structure-activity relationships at the H2 receptor. PMID- 2965341 TI - Using Zoladex in the community. PMID- 2965342 TI - Systemic administration of urocanic acid generates suppression of the delayed type hypersensitivity response to herpes simplex virus in a murine model of infection. AB - Urocanic acid (UCA), the putative photoreceptor for ultraviolet radiation (UV) induced immunosuppression, undergoes a UV-dependent trans to cis isomerization. When UCA, containing a known proportion of the cis isomer, is administered by the intravenous route and live herpes simplex virus is given subcutaneously shortly afterwards, suppression of the delayed type hypersensitivity response to the virus is induced. Two T suppressor cell subsets are generated, one [Ly2+, L3T4-] and the other [Ly2-, L3T4+]. The results are discussed with regard to the differences indicated between local (epidermal) and systemic immunosuppression. PMID- 2965343 TI - [A non-invasive method of blood flow measurement in the arms of patients with arteriovenous fistulas of the Brescia type]. PMID- 2965344 TI - [Atrial natriuretic peptide]. PMID- 2965345 TI - Ectopic pregnancy. Six atypical cases. AB - Improved methods for detecting ectopic pregnancy, including pelvic ultrasound and radioimmunoassay of the beta subunit of human chorionic gonadotropin, have increased the physician's ability to make an early and specific diagnosis. An algorithm presented in this article decreases the chance of misdiagnosis and often defines unusual cases that previously may have been overlooked, such as concomitant intrauterine and tubal pregnancy, bilateral tubal pregnancy, and tubal pregnancy years after bilateral tubal ligation. The algorithm, which remains to be validated, is recommended as an additional tool when the clinical situation is equivocal. PMID- 2965347 TI - [Sciatica of posterior articular origin]. PMID- 2965348 TI - [Renal angiomyolipoma. Diagnostic value of magnetic resonance imaging]. AB - Owing to the wider use of ultrasonography, renal angiomyolipoma is now diagnosed with increasing frequency. Within a few months, 10 female patients were explored by ultrasounds, computerized tomography and nuclear magnetic resonance (Magniscan 5,000, 0.5 Tesla). At ultrasonography the lesion was hyperechogenic in 9 cases and heterogenic in 1 case. At computerized tomography 7 patients had a strongly hypodense lesion and 3 had a lesion of slightly positive density. At magnetic resonance imaging the lesion produced a characteristically loud signal similar to that of fatty tissues. In practice, ultrasonography and computerized tomography combined provide the diagnosis in most cases. In case of doubt, especially since the tumour may be small and asymptomatic and the lesions multiple and/or bilateral, magnetic resonance imaging with its fat-like loud signal confirms the diagnosis and leads to partial resection. When no complications are present, it may even suggest that surgery is unnecessary. PMID- 2965349 TI - [Methods of studying mucociliary function]. AB - The mucociliary system represents the first respiratory tract barrier which acts by removing insoluble particles from the conductive airways. Impairment of mucociliary clearance, due to respiratory mucus and ciliary activity abnormalities, leads to mucus stasis, responsible for infection and bronchial obstruction. In obstructive airway diseases, the efficiency of the mucociliary function can be investigated quantitatively by in vivo and in vitro tests. In vivo mucociliary clearance can be measured by radioactive marker techniques. In vitro tests can evaluate the physical properties and transportability of mucus. The ciliary function can easily be analyzed on ciliated cells collected by nasal or bronchial brushing. Coupled with respiratory function tests, the investigation of the mucociliary function by in vivo and in vitro tests, provides a guideline for selection and assessment of drugs used to correct mucociliary disorders. PMID- 2965346 TI - [Management of a liver tumor disclosed by ultrasonography]. PMID- 2965350 TI - [Crossed reactions in Lyme disease. Value of the Western blot]. PMID- 2965352 TI - [Indications of antibiotherapy in digestive salmonellosis in newborn infants]. PMID- 2965353 TI - [Hemodynamic profile of septic shock in systemic tuberculosis of fulminating course]. PMID- 2965351 TI - [Neonatal shigellosis. A rare cause of rectal bleeding]. PMID- 2965356 TI - [Role of chemotherapy in the treatment of bronchial cancer]. PMID- 2965354 TI - [Cyriac's syndrome. Pseudo-threatening infarction syndrome in a patient with known angina]. PMID- 2965355 TI - [Proposal for the repair of traumatic cavo-suprahepatic lesions]. PMID- 2965357 TI - [Thoughts on ethical problems in neonatal and pediatric resuscitation]. PMID- 2965359 TI - [Elementary test of concentration, orientation and memory. Application to the detection of dementia states in daily practice]. AB - The authors present a french version of the Katzman short orientation memory concentration test. The 6 items of the test include 3 orientation questions, 2 mental control items and an address memory phrase. Time for administration is less than 5 minutes. This french version of the test has been validated as a measure of cognitive impairment in a population of 200 subjects including 140 patients without cognitive impairment and 60 demented subjects. As defined by a 10/11 cut-off score, sensibility and specificity for the diagnosis of dementia were 91 p. 100 and 95 p. 100 respectively. Correlation of the scores with those obtained by the Mini mental state was highly significant. This fast, easy and reliable test seems particularly suitable for the detection of cognitive impairment in clinical practice. PMID- 2965358 TI - [Are retinoids active in the treatment of psoriatic rheumatism?]. PMID- 2965360 TI - [Infection of the newborn infant by the human immunodeficiency virus]. AB - HIV infection of the newborn is now known to result mostly from mother-to-foetus transmission. The risk of transmission is at least 40 p. 100. However, the circumstances of passage are little known, and there is no maternal virological parameter capable of evaluating individual risks. The disease is more severe in children than in adults. Rare are the children who remain asymptomatic for more than 15 months; one out of three of them develop severe acquired immunodeficiency syndrome and die within the first 2 or 3 years of life. A specific encephalopathy is observed in about 30 p. 100 of the infected children. Kaposi's sarcoma is exceptional. Since there is no contagion between children, those who are in fairly good clinical condition should have a family and school life as normal as possible. PMID- 2965361 TI - [Suppression of a subcardial diverticulum of the stomach]. AB - A case of subcardial gastric diverticulum was surgically treated by an original technique which provided good exposure of the lesion and made it possible to sever all adhesions between the diverticulum and adjacent organs. This technique avoids gastrotomy and partial gastric resection. PMID- 2965362 TI - [Isolation of Campylobacter pylori from antropyloric biopsies. A year of systematic research]. PMID- 2965363 TI - [Ileal metastasis of epidermoid carcinoma of the epiglottis. A case]. PMID- 2965364 TI - [Radioimmunologic analysis of antithyroglobulin antibodies in thyroid pathology]. PMID- 2965365 TI - [Significance of the isolation of Stomatococcus mucilaginosus]. PMID- 2965366 TI - [Myoclonic encephalopathy probably attributable to propafenone]. PMID- 2965367 TI - [Thyrotropin anti-receptor antibodies in Basedow disease in remission]. PMID- 2965368 TI - [Autosurveillance of urinary glucose and blood glucose in diabetic children]. PMID- 2965369 TI - [Developing visceral malaria in a patient with chronic HIV]. PMID- 2965370 TI - [Ultrasonic study of gallbladder contraction with ceruletide]. PMID- 2965371 TI - [Prevention of infectious endocarditis in 1987. From theory to practical application]. PMID- 2965372 TI - [The role of transluminal angioplasty in the treatment of renovascular arterial hypertension]. PMID- 2965373 TI - [Effects of delayed-action tetracosactide on chronic pain]. AB - Beta 1-24 corticotrophin was the object of an open trial in the "acute" treatment of chronic pain in 22 patients. The analgesic effect of the drug was assessed before and after two weeks of treatment by means of 3 subjective tests: visual analogue scale, numeric scale and Mac Gill pain questionnaire. The results were statistically significant (P less than 0.01). They confirmed that slow-release tetracosactide exerts a beneficial effect on the sensory-discriminative and affective-emotional components of pain. The drug may be prescribed for short periods (1 to 2 weeks) to patients with chronic pain refractory to the usual treatments. Its analgesic effect is immediate and enables a conventional treatment to be instituted. PMID- 2965374 TI - [Time-related distribution of delayed-action theophylline in the treatment of chronic obstructive bronchopneumopathies]. AB - Three regimens of sustained-release theophylline (Theostat) were administered to 12 male patients with chronic obstructive lung disease in randomized cross over trial. Each 7 days' treatment consisted of: treatment A: 8 mg/kg/day at 07.00 h and 4 mg/kg/day at 19.00 h; treatment B: 6 mg/kg/day at 07.00 h and 6 mg/kg/day at 19.00 h; treatment C: 4 mg/kg/day at 07.00 h and 8 mg/kg/day at 19.00 h. Peak expiratory flow was recorded each day at 07.00, 11.00, 15.00, 19.00 and 23.00 hours, and theophylline plasma levels were determined on the 7th day of each treatment sequence. Cosinor analysis of the data revealed significant circadian rhythms of 24 hr mean Peak expiratory flow for each treatment: the mesor was significantly higher with C and acrophases were located at 14.26 for A, 14.25 for C; a phase shift of the acrophase was detected for B (09.58). These data suggest that an unequal twice daily sustained-release theophylline dosing with higher dose in the evening may be beneficial in the treatment of chronic obstructive lung disease. PMID- 2965376 TI - [Acute granulomatous interstitial nephritis of drug origin]. AB - Epithelioid cell granulomas within the renal interstitium are frequently found in drug-associated acute interstitial nephritis. Their presence is a strong argument in favour of an immunologically mediated nephropathy especially when extrarenal symptoms are absent and if the interstitial infiltrate does not contain eosinophils. They are probably due to a delayed hypersensitivity reaction. Unfortunately, there is no reliable experimental model. The high incidence of permanent renal damage observed in this type of nephritis indicates an early corticosteroid therapy, particularly when renal function does not rapidly improve after withdrawal of the offending drug. PMID- 2965375 TI - [A new treatment of deep venous thrombosis: low molecular weight heparin fractions. Randomized study]. AB - Sixty-eight patients with acute deep vein thrombosis were allocated at random to two treatment groups. One group (n = 33) received a fixed dose of 750 anti-Xa units of a low molecular weight heparin (CY 222 Choay Institute); the other group (n = 35) received standard heparin in doses of 500 IU/kg/24 h. Both treatments were given for 10 days in two daily subcutaneous injections. A second phlebography was performed on the last day of treatment. No haemorrhagic complication was observed in the group treated with CY 222, as opposed to three cases of haemorrhage in the group treated with standard heparin. The initial phlebographic score and the location of deep vein thrombotic lesions were the same in both groups. Angiographic improvement, with more than 30% thrombolysis, was obtained at the end of treatment in 64% of patients in the CY 222 group and in 65% of patients in the standard heparin group (NS). In 2 patients treated with standard heparin the second phlebography showed extension of the thrombosis. The initial score remained unchanged in 1/3 of patients in both groups. The activated partial thromboplastin time was prolonged (2 or 3 fold the normal value) in the standard heparin group and unchanged in the CY 222 group. Anti-Xa activity was significantly higher in the CY 222 group than in the standard heparin group. It is concluded that CY 222 and standard heparin were equally effective in patients with deep vein thrombosis. However, haemorrhagic complications were more frequent with standard heparin that with CY 222. PMID- 2965377 TI - [Treatment of major eventrations. Original use of polyglactin 910 plaques]. AB - An original procedure for treating major incisional hernias is described. A polyglactin 910 plate is hemmed around the edges of the hernia, then tightened by degrees with wide separate stitches until these edges are closely approximated on the midline under low tension, all the forces being distributed over the whole surface of the plate. This procedure can be used alone or associated with another procedure. PMID- 2965378 TI - [Adjustment of tobramycin doses. Lack of reliability of a simple nomogram]. PMID- 2965379 TI - [Toward a moratorium on the use of the Swan-Ganz catheter]. PMID- 2965380 TI - [Decrease of free thyroxine in normal pregnancy]. PMID- 2965381 TI - [Efficacy of plasma exchange in leprotic erythema nodosum]. PMID- 2965382 TI - [Rapid hematological reconstitution after combined reinjection of autologous bone marrow treated by Asta-Z and stem cells from peripheral blood in a patient with Ewing sarcoma]. PMID- 2965383 TI - [Gastric zoster ulcerations]. PMID- 2965384 TI - [A late manifestation of a granuloma on a bronchial suture]. PMID- 2965385 TI - [Syphilis remains a cause of acquired deafness]. PMID- 2965386 TI - [Hemodynamic effects of the administration of nifedipine over an 18-month period in pulmonary arterial hypertension in chronic obstructive bronchopneumopathies]. PMID- 2965387 TI - Chronic Tr-cell lymphoproliferative disease with unusual phenotype: clinical, cytochemical, ultrastructural and immunological studies. PMID- 2965388 TI - Expression of cellular protooncogenes in the mouse male germ line: a distinctive 2.4-kilobase pim-1 transcript is expressed in haploid postmeiotic cells. AB - We report that a 2.4-kilobase (kb) pim-1 transcript is expressed in the germ cells of mouse testis. Analysis of purified populations of spermatogenic cell types indicates that the 2.4-kb transcript is selectively expressed in haploid postmeiotic early spermatids. The evidence for a developmentally regulated expression of pim-1 in haploid spermatids suggests a possible developmental role for this protooncogene product. The 2.4-kb pim-1 transcript present in postmeiotic cells differs in size from the 2.8-kb transcript usually detected in somatic tissues. Similar testis-specific transcripts have been seen for mos and abl genes. These data suggest specificity in transcription or processing of certain genes in haploid male germ cells. We have also analyzed other representative protooncogenes, including examples of protein kinases, the ras family, and the "nuclear" protooncogenes. The results indicate that additional protooncogenes are preferentially expressed in either meiotic pachytene cells or postmeiotic early spermatids. These findings suggest a differential regulation of gene expression in these two developmental stages of germ cells. In particular, analysis of expression of the three members of the ras gene family indicates a distinct temporal differential regulation in the expression of the Harvey, Kirsten, and N-ras genes in these germ cells. PMID- 2965389 TI - Molecular cloning of a human immunoglobulin G Fc receptor. AB - Human IgG Fc receptor (Fc gamma R) cDNA clones were isolated by cross-species hybridization by probing cDNA libraries with the low-affinity Fc gamma R beta 1 cDNA clone from mouse as well as a pool of oligonucleotides constructed from the nucleotide sequence of this Fc gamma R. Three cDNA clones were isolated and analysis of the predicted amino acid sequence indicated that the human Fc gamma R protein is synthesized with a 34-amino acid leader and the mature protein is composed of 281 amino acids. The extracellular region of this Fc gamma R was divided into two domains, which were very similar to each other and to the corresponding regions of both mouse alpha and beta Fc gamma Rs and showed a clear relationship to immunoglobulin variable regions. One possible N-linked glycosylation site was found in each of the extracellular domains. The human Fc gamma R leader sequence was shown to be similar to the mouse alpha Fc gamma R leader sequence, but the transmembrane region was most similar to the mouse beta 1 Fc gamma R. The intracellular domain of the human Fc gamma R was surprisingly different from both mouse Fc gamma Rs. RNA blot analysis of human cells demonstrated two transcripts (2.5 and 1.5 kilobases) that arise by use of different adenylylation signals. The cellular expression of these transcripts suggest that they encode the low-affinity p40 Fc gamma R protein. PMID- 2965390 TI - Alloreactive immune responses of transgenic mice expressing a foreign transplantation antigen in a soluble form. AB - Transfection of cells with the H-2Kk gene lacking the transmembrane and cytoplasmic segments resulted in secretion of the H-2Kk protein, as determined by immunoprecipitation with monoclonal anti-H-2Kk antibodies. Transgenic (H-2b X H 2d)F1 mice were established carrying integrated copies of the modified H-2Kk gene. Expression of the soluble H-2Kk antigen in the transgenic mice was demonstrated in cell supernatants of biosynthetically labeled splenic and thymic Con A blasts as well as bone marrow-derived macrophages. Soluble H-2Kk molecules were also present in the sera of the transgenic animals. No cell-surface expression of the H-2Kk antigen could be observed. In spite of the presence of the soluble H-2Kk molecules in the transgenic mice, the animals were able to generate H-2Kk-specific cytolytic T cells as well as antibody responses when stimulated with cell-surface-bound H-2Kk antigens. These responses were indistinguishable from those of the nontransgenic littermates. Possible explanations for the observed lack of tolerance are discussed. PMID- 2965393 TI - Have you ever had a pain in your back? PMID- 2965391 TI - Protective effects of analogs of luteinizing hormone-releasing hormone against chemotherapy-induced testicular damage in rats. AB - Possible protective effects of analogs of luteinizing hormone-releasing hormone (LH-RH) against testicular damage caused by various cytotoxic agents were investigated in rats. The agonist [D-Trp6]LH-RH (in which Gly-6 is replaced by D tryptophan) and the antagonist N-Ac-[D-Phe(pCl)1,2,D-Trp3,D-Arg6,D-Ala10]LH-RH were administered for 12 weeks: [D-Trp6]LH-RH was given once a month in the form of long-acting microcapsules liberating 25 micrograms of agonist per day, and the antagonist was injected s.c. at a dose of 1000 micrograms per kg of body weight per day for the first 3 weeks and, thereafter, at a dose of 500 micrograms per kg of body weight per day. After a recovery period of 3 months, most seminiferous tubules in the antagonist-treated group showed a normal morphology, while patches of tubules in the agonist-treated group continued to show some suppression of spermatogenesis. Administration of busulfan, cisplatin, or cyclophosphamide produced only a reversible testicular injury, and pretreatment with LH-RH analogs seemed to temporarily enhance the tubular damage. Administration of procarbazine (200 mg per kg of body weight per week for 6 weeks) resulted in decreased testicular weights and increased serum LH levels 1 and 3 months after the discontinuation of treatment. The histology showed severe diffuse damage to seminiferous tubules. The germinal cells completely disappeared and the Sertoli cells were markedly degenerated. This damage was not restored even after a recovery period of 5 months. Some animals were pretreated for 6 weeks with the agonist or antagonist and then received procarbazine for 6 weeks while administration of analogs was continued. In these animals, the decrease in testicular weights and increase in serum LH levels after procarbazine were less marked than in the group not pretreated with the analogs. Three and 5 months after cessation of treatment, a large number of tubules showed a complete restoration of structural morphology in 30-45% of the animals that received procarbazine and the LH-RH agonist or antagonist. These results indicate that pretreatment with LH-RH analogs may protect testes against damage caused by some cytotoxic agents. PMID- 2965392 TI - Lysosomal enzymes in the rat harderian gland are altered by either bromocriptine treatment or hypophysectomy and hormone replacement therapy. AB - Four groups of adult male hypophysectomized rats were injected subcutaneously twice daily between 0800-0900 hr and 1600-1700 hr with either saline diluent, 150 micrograms sheep prolactin and/or growth hormone (GH); intact rats received either saline or 150 micrograms bromocriptine twice daily. After 4 days of treatment, lysosomal enzyme assays revealed significant elevations in both acid phosphatase and alpha-mannosidase enzyme activities in the Harderian glands of saline-injected hypophysectomized rats compared to those in intact controls. beta Glucuronidase levels were depressed and hexosaminidase activity unaffected by hypophysectomy treatment alone compared to intact controls. Lysosomal enzyme activities in hypophysectomized animals treated with prolactin were not different from the hypophysectomized control animals. However, treatment with GH alone or in combination with prolactin had a significant inhibitory effect on beta glucuronidase, hexosaminidase, and alpha-mannosidase enzyme activities in the Harderian gland of hypophysectomized animals. Bromocriptine treatment in intact rats only elevated acid phosphatase activity. In summary, the patterns of responses did not reveal a role for prolactin in the control of Harderian gland lysosomal enzyme activities by the pituitary. However, some of the influence on this target system may be exerted by growth hormone. PMID- 2965394 TI - Behavioral effects of alpha-benzyl-N-methylphenethylamine (ABNMP), a methamphetamine analog: inhibition by ketanserin and para-chlorophenylalanine (PCPA). AB - Acute administration of alpha-benzyl-N-methylphenethylamine (ABNMP) induces lateral heavings, straub tail, backward locomotion, and hindlimb abduction-which are all components of the serotonin (5-HT) syndrome. Ketanserin, a 5-HT-2 receptor antagonist and pretreatment with the 5-HT neurotoxin parachlorophenylalanine (PCPA) attenuated the manifestation of these behavioral abnormalities. These results suggest that ABNMP may cause an acute release of 5 HT similar to that elicited by para-chloroamphetamine. PMID- 2965395 TI - Evidence of correlations between mast-cell histamine and beta-endorphin (ir) from NIL-pituitary in the homeostasis. AB - This study was designed to characterize physiological events related to a single or repeated experimental anaphylactoid reactions (by Compound 48/80) in non stressed or cold-restrained rats. Acute treatment with Compound 48/80 (1 mg/kg, i.p.) increases Beta-endorphin(ir) content in the neurointermediate lobe (NIL) of rat pituitary. Moreover, repeated treatment with Compound 48/80 showed tolerance effects. These animals, exposed to stressful conditions, exhibited a fully evident paw oedema following carrageenin oedema-test, whereas saline-pretreated rats, under the same experimental conditions, showed reduced local inflammation. Since Compound 48/80 produces characteristic, systemic, anaphylactoid reaction in the rat, with a very high degree of selectivity in its action, our results suggest that mast-cell histamine and Beta-endorphin from NIL pituitary are involved in the body's reactivity to stressful stimuli. PMID- 2965396 TI - Rotational behavior induced by 8-hydroxy-DPAT, a putative 5HT-1A agonist, in 6 hydroxydopamine-lesioned rats. AB - Rats with unilateral 6-hydroxydopamine (6-OHDA)-induced lesions of the ascending nigro-striatal pathway have been shown to rotate in response to dopamine (DA) agonists that are not considered to have postsynaptic DA stimulant properties in intact animals, suggesting a relative loss of DA receptor selectivity in the denervated striatum. The present experiments assessed the possibility that this loss of selectivity may extend to serotonin (5HT) agonist drugs. The 5HT-1a agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), at doses of 0.3-3 mg/kg SC, induced robust contralateral rotational behavior (RB) in 6-OHDA lesioned rats that had been preselected on the basis of high responsiveness to the atypical DA agonists 3-PPP and SKF 38393. Rats with unilateral dorsal raphe lesions induced by 5,7-dihydroxytryptamine (5,7-DHT) showed contralateral RB in response to similar doses of 8-OH-DPAT but with a different behavioral pattern. The putative 5HT-1b agonist RU 24969 produced contralateral RB in 5,7-DHT lesioned rats while showing a much weaker effect in 6-OHDA-lesioned rats. Striatal DA levels were depleted by 99% in representative 6-OHDA-lesioned rats but striatal 5HT levels were unaffected. The effects of 8-OH-DPAT in 6-OHDA lesioned rats were therefore not attributable to destruction of ascending 5HT containing neurons. These effects may result from indirect actions, mediated by 5 HT neurons or neuronal receptors, that result from asymmetry of brain DA systems.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965398 TI - Percutaneous transluminal angioplasty of radiation induced renal artery stenosis. PMID- 2965397 TI - Unaltered 5-HT- and desipramine-sensitive [3H]imipramine binding and [3H]5-HT uptake in rat brain after chronic imipramine and norzimeldine treatment. AB - Several reports have shown heterogeneity of [3H]imipramine binding to brain membranes. Recently, a high affinity and 5-HT sensitive [3H]imipramine binding site of protein nature, that was suggested to be identical to the substrate recognition site for 5-HT uptake, was demonstrated. Since most studies on the regulation of the [3H]imipramine binding sites by antidepressants have used desipramine displaceable binding, which is heterogenous in nature and contains binding not related to 5-HT uptake sites, the present report studies the possible effects of chronic (3 weeks) administration of imipramine or norzimeldine (10 mg/kg intraperitoneally twice daily) on 5-HT sensitive [3H]imipramine binding sites. For comparison, desipramine sensitive binding was also studied, as well as the physiological correlate 5-HT uptake. There were no changes in either [3H]imipramine binding or 5-HT uptake after the antidepressant treatment. PMID- 2965399 TI - Effects of adrenalectomy and aldosterone on the action of centrally administered rat atrial natriuretic peptide-(99-126). AB - Intracerebroventricular (i.v.t.) administration of rat atrial natriuretic peptide (99-126) (rANP) increases urinary volume and sodium excretion, but the mechanism is undefined. A diminished mineralocorticoid effect on the kidneys may explain the natriuretic phenomenon. This hypothesis was tested by i.v.t. rANP injection (1.25 micrograms/5 microliters) in conscious, hydrated rats pretreated beforehand with d-aldosterone (20 micrograms/kg, i.p.). Although the absolute amount of sodium excreted was reduced, aldosterone did not affect rANP-induced sodium output at 1 and 3 h. Rats which were sham-operated or bilaterally adrenalectomized (ADX) after four days were pretreated with aldosterone and given an oral water load followed by i.v.t. rANP or saline. In ADX rats natriuresis and diuresis after rANP were still evident. Our results indicate that the natriuretic effect of i.v.t. rANP is unrelated to plasma levels of mineralocorticoids. Likewise, diuresis and natriuresis can occur in the absence of the adrenal glands. PMID- 2965401 TI - Disability and pregnancy: a double dose of disequilibrium. PMID- 2965400 TI - Difficulties in relationships between nonhandicapped and severely mentally retarded children: the effect of physical impairments. AB - Difficulties in relationships between nonhandicapped and severely mentally retarded children (ages 8-12) were examined as a function of the presence or absence of additional physical handicapping conditions. Two sets of data, involving multiple codes of both the handicapped and nonhandicapped children's behavior, were analyzed and aggregated to address this issue. Findings suggested that nonhandicapped children work harder, receive and emit fewer social reinforcers, have less opportunity to play and have fun, and achieve lower levels of social play when their playmate is physically handicapped as well as severely mentally retarded. Implications for the longevity of social relationships with and the community survival of severely multiply handicapped persons are discussed. Strategies for increasing the reinforcement potential of relationships with multiply handicapped children are proposed for future research and implementation. PMID- 2965402 TI - Changes in tissue concentrations of 14C-doxorubicin caused by mitoxantrone, mithramycin A and vinblastine in the rat. AB - The purpose of this study was to determine if mitoxantrone (a new antineoplastic drug) and two other anticancer agents (mithramycin A and vinblastine) could result in changes in the tissue distribution and disposition of doxorubicin in rats. Each of 16 male rats received 1 mg/kg of 14C-labeled doxorubicin on day one and received either saline, or doses of 4 mg/kg of mitoxantrone, 1.7 mg/kg of mithramycin A or 2.9 mg/kg of vinblastine on day three. All rats were sacrificed on day eight. Concentrations of radioactivity in the heart, lung, kidney and muscle were higher in the mitoxantrone treated rats than in the controls. Mithramycin A decreased the concentrations of radioactivity in liver, kidney and fatty tissues, while vinblastine increased the concentrations in heart, liver, lung, muscle and skin. In summary, mithramycin A, vinblastine and mitoxantrone caused a change in tissue concentrations of radioactivity of doxorubicin and/or its metabolites following a single I.V. dose of 14C-labeled doxorubicin in rats. This may cause elevated plasma concentrations of doxorubicin. These findings in the rat could theoretically have implications regarding doxorubicin therapy in man. PMID- 2965403 TI - Lymphocyte dysfunctions and clinical outcome in primary IgA nephropathy. PMID- 2965404 TI - [Thrombolysis and angioplasty at the initial phase of myocardial infarction]. PMID- 2965405 TI - [Interventional cardiology]. PMID- 2965407 TI - Do suppressor T cells exist? PMID- 2965406 TI - [Will inhibition of the hexose monophosphate shunt increase resistance to oxygen deficiency?]. PMID- 2965408 TI - Responsiveness of T cells to mutant major histocompatibility complex class I antigen. I. Obligatory dependence of proliferative response on the presence of stimulator type accessory cells. AB - Proliferative response of splenic T cells of C57BL/6 mice to mutant major histocompatibility complex (MHC) class I antigen (H-2Kbm1) was examined with regard to the role of accessory cells. T cell proliferation in mixed lymphocyte culture (MLC) was not induced when accessory cells were removed from stimulator spleen cells by passage through Sephadex G-10 or nylon-wool column. Anti-Iab antibodies did not inhibit the proliferative response to class I antigen, whereas the same antibodies completely blocked the response to class II antigen (Iabm12). Accessory cells may not be mere presenters of MHC class I antigen because stimulator cells fixed with 0.05% paraformaldehyde lost the stimulating function. The proliferative response was partially recovered by the addition of recombinant interleukin 1 (IL-1) and/or IL-2 to MLC devoid of stimulator type accessory cells. It is concluded that stimulatory type accessory cells were obligatorily involved in the T cell proliferation, and the production of IL-1 by accessory cells is thought to play a critical role in this process. PMID- 2965409 TI - Influence of continuous luteinizing hormone-releasing hormone agonist treatment on steroidogenic enzymes in the human testis. AB - Nine elderly men with prostatic carcinoma underwent treatment with a LHRH-agonist (Zoladex, ICI) for 3-6 months. At the end of the treatment period the patients underwent subcapsular orchidectomy. Testicular tissue was incubated with different tritiated testosterone precursors. Conversion mediated by several testicular steroidogenic enzymes was compared between Zoladex-treated patients and nineteen non-treated patients who underwent orchidectomy because of prostatic carcinoma. Serum concentrations of LH, FSH and testosterone were determined before and during treatment in the treated patients. The LHRH agonist treatment induced significantly decreased conversion mediated by the enzymes 3 beta hydroxysteroid dehydrogenase, 17 alpha-hydroxylase and C17-20 lyase. Conversion mediated by 17 beta-ketosteroid reductase was also decreased although not as dramatically as the other enzymes, while conversion mediated by 20 alpha dehydrogenase was increased. Serum concentrations of testosterone decreased to castration levels. Serum gonadotrophins decreased but remained within normal levels suggesting that "desensitization" at the pituitary level was not the only mechanism of action of the LHRH-agonist. PMID- 2965410 TI - Results of abdominal levator-muscle repair in urinary stress incontinence. AB - Results of the abdominal levator-muscle repair in 175 stress incontinent women are presented. Two years after the operation 83% were cured or improved regarding urinary incontinence. The urodynamic evaluation revealed a normalization of the maximum flow rate, residual urine and cystometry in nearly all cases. The abdominal levator-muscle repair carried a low morbidity, and is especially recommended in women with combined urinary and anal incontinence. PMID- 2965411 TI - [Experience in teaching problems of prevention in the medical school]. PMID- 2965412 TI - [Diagnosis of rare cases of Q fever in the northwestern region]. PMID- 2965413 TI - [Public health in the Russian Federation (on the 70th anniversary of Great October]. PMID- 2965414 TI - [Development of pharmacy in the RSFSR during the past 70 years]. PMID- 2965415 TI - [Clinical lectures at a medical institute]. PMID- 2965416 TI - [Improvement in specialized care for burns in the RSFSR]. PMID- 2965418 TI - [Role of immunoregulatory cells in supporting recovery processes]. PMID- 2965417 TI - [Primary pulmonary hypertension]. PMID- 2965420 TI - [Emergency care in specialized departments of general hospitals]. PMID- 2965419 TI - [Information support of research in a general clinical institute]. PMID- 2965421 TI - [Optimization of the consultative polyclinic care in a specialized department]. PMID- 2965422 TI - The relationship between anthropometric parameters and measurements of the human inguinal region. AB - Several reports suggest a definite relationship between inguinal hernia and pelvic measurements. In order to assess this opinion, measurements of the inguinal region, in particular of the inguinal triangle (Hesselbach) were statistically compared with 14 different anthropometric parameters in 73 white human male corpses. The correlation tests between the inguinal and anthropometric parameters showed very low r values, generally being null to three decimal places. The importance and possible etiologic role of these findings are discussed. PMID- 2965424 TI - Durability of confluent endothelial cell monolayers on small-caliber vascular prostheses in vitro. AB - Since implantation of small-caliber vascular grafts with preformed confluent endothelial cell monolayers (ECMs) may prevent acute platelet deposition and thrombus formation, we have evaluated the conditions necessary to produce durable ECMs. Cultured human umbilical-vein ECs in buffer were attached to vascular grafts--either expanded polytetrafluoroethylene (ePTFE) or knitted Dacron, both with inside diameters of 4 mm--precoated with collaten type I and perfused in vitro with serum-containing culture medium to achieve cell spreading into confluent monolayers. The number of cells attached was quantified by DNA measurements and indium-111 labeling. Morphology was evaluated by scanning electron microscopy. The maximum number of cells that attached acutely was 3.6 X 10(5) cells/cm2 graft, and the minimum number needed for confluence was 1.4 X 10(5) cells/cm2 graft. Confluence was morphologically complete after 2 hours of in vitro perfusion at 15 ml/min. When ECMs were exposed to varying flow rates, cell retention after 1 hour was 96.1% +/- 5.6% at 50 ml/min, 94.6% +/- 6.1% at 100 ml/min, 77.6% +/- 10.8% at 200 ml/min (p less than 0.001), and 40.1% +/- 10.4% at 400 ml/min (p less than 0.0001). Confluence was maintained for all grafts exposed to flows of 100 ml/min or less. Fewer cells were retained when acutely attached, unspread cells (71.5% +/- 11.5%) were compared with established ECMs (89.9% +/- 6.7%) at a flow rate of 100 ml/min (p less than 0.0001). ECMs on knitted Dacron were more durable than on ePTFE (82.7% +/- 5.3% versus 75.5% +/- 4.8% remained attached at 200 ml/min, P less than 0.05). 111Indium-labeled and unlabeled cells were equivalent with respect to saturation level attachment, spreading time to confluence, and durability under flow at 100 ml/min. We conclude that confluent and durable endothelial cell monolayers can be established on small-caliber vascular grafts within 2 hours. PMID- 2965423 TI - Gonococcal keratoconjunctivitis. AB - Gonococcal keratoconjunctivitis is a potentially devastating infection, because Neisseria gonorrhoeae can cause a rapid, severe, ulcerative keratitis resulting in visual loss. The therapeutic decision making process is complicated by the necessity for prompt, effective parenteral therapy, frequent coinfection with other sexually transmitted diseases, and emergence of antibiotic resistance. Because of the evolving problem of antibiotic resistance and the need for cost containment, the current recommendations of hospitalization for intravenous penicillin may need to be modified. The third generation cephalosporin, ceftriaxone, has properties that suggest it may be the best available antimicrobial agent as a single-dose treatment of gonococcal conjunctivitis. Spectinomycin may be a useful alternative in the penicillin-allergic adult patient. PMID- 2965425 TI - Effect of inhaled formyl-methionyl-leucyl-phenylalanine on airway function. AB - Formyl-methionyl-leucyl-phenylalanine (FMLP), a synthetic, acylated tripeptide analogous to bacterial chemotactic factors, has been shown to cause bronchoconstriction in guinea pig, rabbit, and human airways in vitro. To determine whether FMLP causes bronchoconstriction in man in vivo, a preliminary study was undertaken in which five non-smokers (mean age 35 years, FEV1 94% (SEM 5%) predicted) and five smokers (mean age 34 years, FEV1 93% (6%) predicted) inhaled aerosols of FMLP. None of the subjects showed airway hyperresponsiveness to histamine (the provocative concentrations of histamine causing a fall of greater than or equal to 20% in FEV1 (PC20) were over 8 mg/ml). FMLP dissolved in 50% dimethylsulphoxide and 50% saline in concentrations of 0, 0.06, 0.12, 0.25, 0.5, 1.0, 2.0, and 4.0 mg/ml was administered to the subjects by means of a French-Rosenthal dosimeter, FEV1 being recorded after inhalation of each concentration. Dose dependent falls in FEV1 occurred in five non-smokers (geometric mean 1.76, 95% confidence limits 0.87-3.53 mg/ml) and three smokers (0.23, 0.07-0.78 mg/ml), with two smokers not responding by 20% to the highest concentration of FMLP. On a separate day the FMLP dose-response curves were repeated after nebulisation of 500 micrograms of ipratropium bromide. The PC20 FMLP in the responders more than doubled. In six additional normal subjects a histamine inhalation test was performed before and four and 24 hours after inhalation of FMLP. All subjects remained unresponsive to histamine. These results show that FMLP is a potent bronchoconstrictor in some non-asthmatic individuals in vivo and this may be important in bronchoconstriction related to infection in patients with chronic obstructive lung disease. PMID- 2965426 TI - Expression of the atrial natriuretic peptide gene in the cardiac muscle of rat extrapulmonary and intrapulmonary veins. AB - Atrial natriuretic peptide is a peptide regulating salt and water balance, originally isolated from the cardiac atrium, where it is synthesised as part of a precursor molecule in specialised myocardial cells. The myocardium extends into the extrapulmonary part of the pulmonary veins in many species, including man. In some small mammals, however, such as the rat, mouse, and bat, it extends further to veins in the peripheral parts of the lung. Since this myocardial layer is continuous with that in the atrium, we have looked for the possible expression of the atrial natriuretic peptide gene in this tissue in rats. Strong immunoreactivity was seen for both the peptide and the N terminal sequence (cardiodilatin) of its precursor in extrapulmonary veins and in intrapulmonary veins extending into the lung as far as the second branching point, where it was localised in the dense cored granules by electron microscopy; in situ hybridisation showed atrial natriuretic peptide messenger RNA at identical sites. Chromatography and radioimmunoassay of extracts of extrapulmonary and intrapulmonary veins showed most of the atrial natriuretic peptide immunoreactivity to be in the uncleaved (precursor molecule) form. Thus the peptide is synthesised in veins both outside and inside the lung, and these extra atrial sites may be an important additional source of circulating atrial natriuretic peptide. PMID- 2965427 TI - Severe deep vein thrombosis in a 2-year-old child with protein S deficiency. PMID- 2965428 TI - Immunoradiometric assay for the calcium-stabilized conformation of human protein S. AB - Rabbit polyclonal anti-protein S serum was fractionated with immobilized human protein S to establish solid-phase immunoradiometric assays recognizing Ca(II) dependent and NonCa(II)-dependent epitopes of human protein S. The two assays were specific for PS:Ca(II)Ag and PS:NonCa(II)Ag and highly sensitive with a lower limit of detection of about 2.5 ng/ml. PS:Ca(II)Ag and PS:NonCa(II)Ag levels were measured in immunopurified protein S, thrombin-modified protein S and chymotrypsin-cleaved protein S. Only in chymotrypsin-cleaved protein S an important discrepancy between the two antigen levels was observed. Ranges for the concentration of PS:Ca(II)Ag and PS:Non-Ca(II)Ag and their ratio were established in plasma of healthy individuals (0.92 +/- 0.13 U/ml, 0.98 +/- 0.21 U/ml, 0.96 +/ 0.17, respectively). In a group of patients using oral anticoagulant therapy the ratio PS:Ca(II)Ag/PS:NonCa(II)Ag decreased at increasing intensity of anticoagulation suggesting the presence of sub- and noncarboxylated protein S molecules. In plasma of patients with a hereditary type I protein S deficiency PS:Ca(II)Ag and PS:NonCa(II)Ag were reduced to the same extent: mean ratio 1.02 +/- 0.12 in the group not on oral anticoagulant treatment and 0.94 +/- 0.10 in the group on oral anticoagulant therapy. Analysis of patients with a history of unexplained thrombo-embolic disease did not reveal individual patients with a PS:Ca(II)Ag/PS:NonCa(II)Ag ration below the lower limit of the normal range (mean ratio 1.05 +/- 0.17), suggesting that the frequency of genetic protein S variants with defects in the Ca(II)-stabilized conformation is very low. PMID- 2965429 TI - A functional test for protein S activity in plasma. AB - The physiological role of coagulation cofactor Protein S (PrS) for activated Protein C (APC) has recently been appreciated by the description of patients with PrS-deficiency, suffering from thromboembolism. The present study introduces a one-stage clotting assay for the assessment of PrS functional activity in plasma samples. The assay procedure is based on a factor Xa-initiated clotting test utilizing a mixture of AL(OH)3-adsorbed substrate plasma and patient's plasma supplemented with purified prothrombin (0.15 microM) and APC (0.05 microM), with phospholipids and CaCl2. Owing to the varying concentration of PrS in the sample plasma, clotting times were prolonged up to 25 seconds in the presence of APC, whereas no prolongation occurred in its absence. The test procedure proved to be specific for PrS, since preincubation with monospecific antibodies against PrS abolished the prolongation of clotting time, while reconstitution of adsorbed plasma with purified PrS restored its cofactor activity completely. The functional assay showed an inter-assay and intra-assay variation in the normal range of 11.7% and 10.1%, respectively (n = 20). PrS activity in a group of unselected patients (n = 34), revealing no abnormalities in global coagulation tests, amounted to 95.8 +/- 16.5% (mean +/- S.D.) with a range from 67% to 136% when analyzed in comparison to a plasma pool constituted from healthy volunteers. Patients (n = 32) undergoing oral anticoagulant therapy presented 21.1 +/- 10.8% residual PrS-activity accompanied by a concomitant decrease in PrS-antigen levels to 69.9 +/- 21.2%. The assay described is sensitive, it can be performed on routine basis and allows the detection of patients with PrS-deficiency. PMID- 2965430 TI - Inhibition of human factor VIIa-tissue factor activity by placental anticoagulant protein. AB - Previous studies indicated that human placental anticoagulant protein, a member of the lipocortin family, prolonged the clotting time of normal plasma when clotting was induced by brain thromboplastin or by kaolin in the presence of cephalin and calcium. Using a two-stage amidolytic assay to assess factor X activation and a tritiated peptide release assay to assess factor IX activation, we have examined the ability of purified preparations of placental anticoagulant protein (Mr = 36.5 kDa) to inhibit the activation of either factor X or factor IX by a complex of human factor VIIa-tissue factor. Placental anticoagulant protein markedly inhibits factor X and factor IX activation by factor VIIa-tissue factor in a non-competitive manner with Ki values of 40 nM and 70 nM, respectively. Placental anticoagulant protein had no effect on factor Xa amidolytic activity, and its inhibitory activity was not diminished by prior incubation with antibody raised against partially purified plasma extrinsic pathway inhibitor. Binding of placental anticoagulant protein to phospholipid vesicles, crude tissue factor and purified, relipidated human brain tissue factor apoprotein was observed only in the presence of calcium ions. These results indicate that placental anticoagulant protein is a potent factor VIIa-tissue factor inhibitor and suggests that its mechanism of action involves binding to the phospholipid portion of the tissue factor lipoprotein. PMID- 2965432 TI - [Postpartum backache. An epidemiological study]. PMID- 2965431 TI - [Chronic backache. Diagnosis and dysfunction]. PMID- 2965433 TI - [Gonadotropin releasing hormone agonists. A new therapeutic principle in gynecology]. PMID- 2965435 TI - [Thyroid function in Down's syndrome]. PMID- 2965434 TI - [Atrial natriuretic peptide. A review]. PMID- 2965436 TI - [Compensation practice of labor unions in diseases caused by ionizing radiation]. PMID- 2965437 TI - [Professional and ethical responsibilities of dentists to HIV-positive patients and patients with AIDS]. PMID- 2965438 TI - Reinfection of people with Ascaris lumbricoides following single, 6-month and 12 month interval mass chemotherapy in Okpo village, rural Burma. AB - A longitudinal study on reinfection with Ascaris lumbricoides was continued on a random sample of 50% of the infected population following a horizontal cross sectional study in Okpo village, near Rangoon. The study sample was again randomly divided into two subsamples, the six-month interval worming group and the 12-month interval worming group. Microscopic examination of stool for Ascaris eggs on the 7th day and 30th day, combined with counting eggs and worming with levamisole, were carried out at two successive six-month intervals on the first subsample. Stools were examined and eggs counted on the 7th day and then monthly up to 12 months, followed by worming, in the second subsample. The worms expelled in the first 72 hours after treatment were counted. The findings over a 12-month follow-up period after one treatment included: mean monthly incidence of 20%, higher and more rapid return to previous prevalence and intensity of infection in children or 'wormy' persons than in adults or 'non-wormy' persons. Six-monthly chemotherapy definitely reduced intensity of infection in the children and adults whereas 12-monthly treatment lowered intensity in adults only. Predisposition to acquisition of high or low intensity of infection was also observed. Other findings and the implications of this study for strategies for control of ascariasis are discussed. PMID- 2965439 TI - Age and sex differences in new and recurrent cases of guinea worm disease in Nigeria. AB - In this population-based cross-sectional survey of the prevalence and incidence of guinea worm disease in Idere, a rural agricultural community of Oyo state in Nigeria, epidemiological data were collected by household interview of all 501 households (6527 persons, 3594 females and 2933 males). 86% of the households had at least one case. The prevalence was 32.4% in the overall population, but varied markedly by age and sex. About 10% of prevalent cases were first-time infections or new cases. Males over 19 years of age had a higher prevalence rate than adult females; however, females had a higher prevalence rate at younger ages. The prevalence in females peaked (47%) at 35 to 44 years; for males the proportion was highest (57%) at ages 45 to 54 years. PMID- 2965440 TI - Bulk cryopreservation of lymphocytes in glycerol. AB - The authors describe a method for freezing large amounts of peripheral blood lymphocytes (PBL) in a 20 percent glycerol solution. Between 0.6 and 4.3 X 10(9) cells in autologous plasma were frozen in polyethylene freezing bags in a final volume of 50 ml. The recovery after thawing averaged 89 +/- 14 percent with a mean viability by trypan blue dye exclusion of 80 +/- 7 percent (n = 11). In aliquots of fresh and frozen-thawed PBL from the same subjects radiolabeled with 111In, the radiolabeling efficiency for both fresh and thawed cells was 49 +/- 15 percent (p = 0.98, n = 5). The mitogen mean stimulation indices for glycerol frozen cells (471 with phytohemagglutinin-M, 176 with pokeweed mitogen, and 380 with concanavalin A) were superior to those of cells frozen by a standard technique with dimethylsulfoxide (DMSO) (141, 47, and 123; p less than 0.05) and comparable to those of fresh PBL (403, 75, and 147). In a mixed lymphocyte culture, glycerol-frozen PBL showed significantly greater responsiveness to a pool of stimulator cells than did PBL frozen in DMSO (p = 0.03). Thawed cells are viable and functional as demonstrated by their response to mitogens and their ability to stimulate and respond in mixed lymphocyte culture. PMID- 2965441 TI - Fowl cholera in quail in Burma. PMID- 2965442 TI - Chromosomal aberrations in mildly mentally retarded children in a northern Swedish county. AB - Few studies of the frequency of chromosomal aberrations in an unselected material of mildly mentally retarded children have been published. The present paper summarizes the chromosomal abnormalities in children with mild mental retardation (MMR) in Vasterbotten in the northernmost part of Sweden. Chromosome analyses were carried out by routine methods. In addition, children whose MMR had no clear diagnosis or cause were investigated as regarding X-chromosomes with a fragile site and selected cases with banding techniques. Every mentally retarded child born between 1959 and 1970 in the county of Vasterbotten was traced. Out of a total number of 40,871 individuals, 171, i.e. 4.2 per 1,000, were found to be mildly mentally retarded. Chromosomal aberrations were seen in 11.9% of the children with MMR, compared with 39.1% of the 161 children with severe mental retardation (SMR) in the same population. The proportion of cases with mental retardation of unknown etiology is high, especially amongst those with MMR. The use of high resolution banding and other modern cytogenetic methods should reduce this figure. PMID- 2965443 TI - [Diseases of the organ of vision in children (data from a laboratory of pathological anatomy)]. PMID- 2965444 TI - [Upper respiratory tract morbidity with temporary loss of work capacity in agricultural workers]. PMID- 2965447 TI - [Clinical picture and treatment of lumbosacral ligamentous pain of microtrauma origin]. PMID- 2965445 TI - [Comparison of the effects of tercleucine and tryptophan superanalogs of LH-RH on gonadotropin secretion in heifers]. AB - In six heifers without sexual organs the effects were compared of the superanalogues LH-RH [(D-Tle6) and (D-Trp6)] on the secretion of luteinizing hormone (LH) and follicles of stimulating hormone (FSH). The superanalogues were given i. m. at dosages of 10, 50 and 100 micrograms pro toto. Dosages of 50 and 100 micrograms of both superanalogues had a greater effect on the secretion of LH than the dosage of 100 micrograms. The highest average concentration of LH during the time of application (D-Tle6) was recorded in the dosage of 10 micrograms (7.84 ng.ml-1). The average concentrations of LH after application (D-Trp6) were within the range of 5.46 to 9.29 ng.ml-1. Because of the great variability of the concentrations of LH no significant differences were ascertained due to the influence of dosage. From a comparison of the concentrations of LH with the same dosage it emerged that (D-Trp6) after an application of 50 micrograms significantly increased the concentration of LH from the 60th to the 240th minute and thus had a more protracted effect. With dosages of 10 and 100 micrograms of the superanalogues no statistically significant differences were recorded after two hours from application. With (D-Trp6) the higher stimulative effect on the secretion of LH was statistically confirmed. The highest concentrations of follicles of stimulation hormone (FSH) (204.6 and 228.6 ng.ml-1) were found from the 40th to the 100th minute after the application of dosages of 50 and 100 micrograms (D-Tle6). The protracted effect was greatest with a dosage of 100 micrograms (270 mins). (D-Trp6) at a dosage of 100 micrograms caused the greatest effect on the secretion of FSH (226.8 ng.ml-1 for a period of 180 mins). The lower dosages of analogues scarcely differed in response. The dosage of superanalogues has an influence on the concentration of FSH in peripheral blood and on the duration of the protracted effect. PMID- 2965446 TI - The phenomenon of stress: concepts and mechanisms associated with stress-induced responses of the neuroendocrine system. AB - 'Stress', 'stressors' and the 'stress response' are terms which represent concepts rather than being precisely definable. No single theory of stress has been universally accepted. Selye's General Adaptation Syndrome of stress remains the primary theory for the basis of stress research in food-producing animals; however, other concepts and theories have been developed. This paper reviews current theories and suggests that the complexity of research on stress in food producing animals can be attributed to the lack of concrete theories and concepts regarding identifiable stressors, the endocrine responses stimulated by potential stressors and the complex and integrated biological changes in the neuro endocrine system resulting from exposure to hypothesized stressors. PMID- 2965448 TI - In vivo evolution of adenovirus 2-transformed cell virulence associated with altered E1A gene function. AB - Neoplastic cell populations may evolve to a state of higher virulence in immunocompetent hosts. Transforming gene involvement in this process of tumor progression was evaluated using adenovirus type 2 (Ad2)-transformed hamster cells that are highly susceptible to destruction by natural killer cells and activated macrophages, due to Ad E1A gene function, and are nontumorigenic in immunocompetent animals. Cells selected for increased tumorigenicity retained parental cell patterns of viral gene integration and methylation and expressed Ad2 E1A proteins but exhibited altered E1A function evidenced by decreased susceptibility to killer cell-mediated lysis and inability to support E1A(-) mutant virus replication. The data suggest that an interruption in cellular pathways of E1A expression may result in increased transformed cell virulence. PMID- 2965449 TI - Use of deletion and point mutants spanning the coding region of the adenovirus 5 E1A gene to define a domain that is essential for transcriptional activation. AB - To help in identifying functional domains within Ad5 E1A proteins, we have constructed a series of mutants that create deletions throughout these products. We have also produced several mis-sense point mutations in the unique 13 S mRNA region. These mutated E1A regions have been tested in plasmid form for their ability to activate transcription of an E3-promoted CAT gene. From the results, a major domain for transactivation has been identified. This begins between residues 138 and 147, ends between residues 188 and 204, and encompasses the unique 13 S region. This domain is sensitive to mis-sense mutations. Transactivation was unaffected by small deletions in the N-terminal half of E1A proteins between residues 4 and 138, but was destroyed when this whole region was deleted. The C-terminal 71 residues may affect transactivation, but the results with the mutant in which this region was deleted were variable. The results obtained with these mutants are discussed in relation to the transactivation obtained by J. W. Lillie et al. [(1987). Cell 50, 1091-1100] with a synthetic peptide similar to the domain described here. PMID- 2965450 TI - A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. AB - Early region 1 (E1) of the human adenoviruses has many intriguing properties which have prompted numerous mutational studies to help delineate and characterize the domains responsible for these functions. In mutational analyses being done currently, the E1 region is usually cloned into a bacterial plasmid where it is mutated and then the altered E1 sequences are "rescued" back into infectious virus. The most frequently used rescue procedures are somewhat tedious, requiring the purification and fractionation of linear viral DNA or DNA fragments, and often involve the screening of numerous plaque isolates. Several observations we have made recently on the properties of adenovirus DNA in infected cells and on infectious plasmids in transfected cells led us to design a new approach for rescuing E1 mutations into infectious viral genomes. We constructed a plasmid, pJM17, containing the entire Ad5 DNA molecule, with an insert in the E1 region that exceeds the packaging constraints of the adenovirus capsid. Following transfection of pJM17 into 293 cells the plasmid DNA is able to replicate but cannot be packaged into infectious virions. In contrast cotransfection of 293 cells with pJM17 plus an E1-containing plasmid carrying mutated sequences produces recombinant virions at high efficiencies. Neither plasmid needs to be linearized prior to contransfection. The technique eliminates the need to purify and manipulate infectious virion DNA and since no unique restriction sites are needed, both E1A and E1B mutants' as well as foreign gene inserts in the E1 region can be easily rescued into virus. PMID- 2965452 TI - [Development of physiotherapy and health resort treatment in Byelorussia (1917 1987)]. PMID- 2965453 TI - [Regional hemodynamics in patients with osteoarthrosis as affected by treatment at the Tskhaltubsko mineral baths]. PMID- 2965451 TI - [Doppler measurement of pressure gradients in mechanical heart valves and color coded mapping of their hemodynamics]. PMID- 2965454 TI - [Operations management of balneotherapy of patients at the Pyatigorsk health resort using a computer]. PMID- 2965457 TI - [Assessment instrument for the evaluation of health care needs of elderly citizens]. AB - It's necessary for registration of care-work to evolve a diagnostic instrument, that ascertains the individual needs. This instrument has to consider physical, physical and social components of capacity. A procedure is introduced, that is intended for the work of social workers and at once its's unitable for the investigation of essential determining factors and their mutual dependency. PMID- 2965456 TI - [Responsibilities of the psychologist in the management of elderly and disabled persons]. AB - After a short introduction to the general objectives of care for elderly people the special tasks of psychologists in old people's homes are discussed. Possibilities and limitations of their work are indicated. PMID- 2965455 TI - [Incomplete uterine perforation diagnosed by laparoscopic examination]. PMID- 2965458 TI - [Acne therapy with isotretinoin in dermatologic practice. A multicenter phase IV study of 788 patients]. AB - In a multicenter study, 788 patients suffering from severe acne resistant to therapy were treated with isotretinoin by 758 dermatologists in private practices. The study was conducted using standardized test forms and questionnaires. The goal of the study was to establish the risk/benefit ratio of the drug, if it was applied to patients of a dermatological practice. The initial daily dose amounted to 0.5 mg/kg body weight, the time of treatment was 12 to 16 weeks. The clinical efficacy was judged very good or good by 90% of the doctors and patients. The kind, frequency, and intensity of the side effects observed corresponded with those found in previous studies conducted at hospitals. Our critical analysis of the results left no doubt about the therapeutic value and the safety of isotretinoin, if it is used by dermatologists in private practices. PMID- 2965459 TI - [Disorder of hair growth in hyperprolactinemia]. AB - Functional hyperprolactinemia was found in five female patients, 25-35 years old, seeking medical consultation for hair loss, together with hypertrichosis (4x), disturbances of cyclic bleeding periods (4x), secondary amenorrhea (2x), galactorrhea (2x), seborrhea (2x) and persisting acne (1x). Other hormonal parameters including testosterone levels and thyroid gland function tests were unchanged. Prolactinoma was excluded by x-ray diagrams, partly also by computer tomograms of the sella. In two patients increased telogen effluvium was found by trichogram examination with some dystrophic hairs; in one patient only dystrophic hairs were seen, whereas, in two cases, hair loss was not present at the time of our clinical examination. These observations indicate that cutaneous symptoms such as seborrhea, acne, hypertrichosis/hirsutism, alopecia(= SAHA syndrome) may evidently occur in hyperprolactinemia, representing or mimicking androgen-induced skin symptoms. In such cases, therefore, evaluation of prolactin levels together with androgen blood levels and thyroid gland function tests should be performed to exclude underlying endocrinopathy. PMID- 2965460 TI - Androgenic influence on the activity of the uropygial gland: the effect of dehydroepiandrosterone and androstenedione treatment on the histodynamics and the secretory lipids of the uropygial gland of pigeons. PMID- 2965461 TI - Prevention of horizontal transmission of hepatitis B: efficacy of hepatitis B immunoglobulin and vaccine in an institution for the handicapped. AB - In a Japanese institution for the handicapped with confirmed continuous outbreaks of hepatitis B virus (HBV) infection by horizontal nosocomial transmission, 29 susceptible subjects (8 institutionalized children and 21 medical staff) were injected intramuscularly with anti-human HBs immunoglobulin (HB Ig) and subcutaneously with HB vaccine. All cases acquired HBs antibody after injection of HB Ig and seropositivity for HB after the third inoculation of HB vaccine was 78.6%. No new case of HB occurred among the study population throughout the period investigated. This suggested the effectiveness of HB Ig and HB vaccine in the prevention of horizontal nosocomial transmission of HBV. PMID- 2965462 TI - Low dose intradermal vaccination against hepatitis B in mentally retarded patients. AB - Ninety-two patients and 28 staff members of an institute for mentally retarded patients were immunized intradermally with 2 microgram doses hepatitis B vaccine (HB-Vax), initially at month 0 and 1, and after evaluation followed by two booster injections at month 5 and 10, respectively. Blood samples were taken at months 2 and 11. Seroconversion rates of the two groups were similar on both occasions. Titre distributions for mentally retarded patients and staff members corrected for age, side effects and sex using multiple regression were significantly different at month 2 (p = 0.05) but not at month 11. Geometric mean titres for mentally retarded patients and staff members were 39 and 67 at month 2, respectively, and 1082 and 672 IU l-1 at month 11, respectively. Percentage anti-HBs/a(w) differed significantly between patients and staff members. Side effects, largely local (pigmentation), were observed with a frequency of about 35% in both mentally retarded patients and staff members. Since 92% of the mentally retarded patients had anti-HBs titres greater than 10 IU l-1 and approximately equal to 80% greater than 100 IU l-1, low dose vaccination, four injections by intradermal route, seems to be a suitable cost-reducing immunization system in institutes for mentally retarded patients. PMID- 2965463 TI - [The processes of the formation of immunological tolerance in the adult organism]. PMID- 2965464 TI - [Diagnostic characteristics of congestive right-ventricular heart failure in chronic bronchitis under alpine conditions]. PMID- 2965465 TI - [Optimization of the diagnostic and treatment methods in chronic heart failure and the characteristics of its pharmacotherapy under alpine conditions]. PMID- 2965466 TI - [Results and prospects for the development in the Byelorussian SSR of comprehensive research in oncology]. PMID- 2965467 TI - [Summing up the results of carrying out the Republican Complex Program of Fundamental Research in Medicine in the Byelorussian SSR (1980-1985) and outlining the 1986-1990 program in the light of decisions of the 27th Congress of the Communist Party of the Soviet Union (CPSU)]. PMID- 2965468 TI - Inhibition of vaccinia virus replication by nicotinamide: evidence for ADP ribosylation of viral proteins. AB - Replication of vaccinia virus (VV) in monolayers of BSC40 cells was inhibited 99.9% in the presence of 60 mM nicotinamide (NIC), a competitive inhibitor of ADP ribosylation reactions. Dot-blot hybridization analysis of infected cell extracts utilizing a VV DNA-specific probe indicated that the drug had only minimal effects on viral DNA synthesis. SDS-polyacrylamide gel electrophoresis of newly synthesized VV proteins pulse-labeled at early (2 h) or late (8 h) times post infection revealed that although the full spectrum of expected viral polypeptides was evident, quantitative differences in the levels of expression of a distinct subset of viral proteins were observed in the presence of the drug. Velocity sedimentation of virus-infected cell lysates established that no mature particles were assembled in drug treated cells. Additional evidence suggesting that VV morphogenesis was abortive in the presence of NIC was obtained by pulse-chase labeling experiments that demonstrated that the two VV major late core polypeptide precursors P94 and P65, whose proteolytic processing to VP62 and VP60 is intimately associated with viral assembly, were not cleaved in the presence of NIC. Interestingly, growth of VV in the presence of [3H]adenosine resulted in the metabolic labeling of eight proteins that were associated with purified virions. These proteins co-migrated with proteins labeled with [3H]adenosine that were present in extracts of VV-infected, but not uninfected, cells. These analyses also revealed that the [3H]adenosine-labeling of a subset of cellular proteins (MW 18-20 kDa, possibly histones) was increased 4-fold by VV infection. The observed induction of either increased synthesis or hyper-modification of these 18-20 kDa proteins was inhibited by NIC. These results are discussed with respect to whether one or more VV polypeptides are subject to obligatory ADP-ribosylation modification reactions in order to attain their active configuration, and if so, whether the enzymes catalyzing these reactions are specified by the virus or host cell. PMID- 2965469 TI - [Permanent dentition--a longitudinal study of 250 schoolchildren between 7 and 15 years of age]. PMID- 2965470 TI - [X-ray follow-up on the progress of initial proximal caries in adolescents]. PMID- 2965471 TI - [Clinical value of sound phenomena of the temporomandibular joint]. PMID- 2965472 TI - [The importance of the secretory defense of the oral cavity for the etiopathogenesis of alveolitis]. PMID- 2965473 TI - [Local anesthesia and hypertension]. PMID- 2965474 TI - [Plaque microflora of healthy gingiva as well as gingivitis and marginal periodontitis]. PMID- 2965475 TI - [Teratogenically modified palatogenesis in the Uje:WIST rat. 3. Proliferative activity in the mesenchyme and epithelium of the palatine process]. PMID- 2965476 TI - [Incidence of initial caries and structural anomalies of the dental enamel in patient with cheilo-gnatho-palatoschisis]. PMID- 2965477 TI - [Tracheotomy in maxillofacial surgery]. PMID- 2965478 TI - [Detection of immune complexes in intrauterine retardation]. AB - We tried to find an additional diagnostic parameter for intrauterine growth retardation. The immune complex estimations were carried out by means of precipitation of immune complexes with Polyethylenglykol 6.000 in 203 primi and multiparae suspected for intrauterine growth retardation. The control group (y = 0.073 X + 0.372; r = 0.459) showed an increase of immune complex concentrations towards the end of pregnancy. A possibility for interpretation is the daily penetration of fetal material in to the maternal circulation. Women with suspicion of intrauterine growth retardation (y = -0.045 X + 1.61; r = -0.053) showed a decrease of immune complex concentration at the end of pregnancy. This course is contrary to that one of immune complex concentrations in the control group. The distribution of immunglobulins in the precipitate is corresponding to the proportions in serum. We found mainly IgG and IgM, but also high molecular serumproteins as Alpha-2-Makroglobulin and Orosomucoid. A discreet correlation between intrauterine growth retardation and pathological immune complex values may be present. PMID- 2965480 TI - [Percutaneous thermocoagulation of sensory lumbar nerve roots in the treatment of pain]. AB - The study was carried out on the experiences with 20 patients with low back pain. They were treated by thermoelectrocoagulation of the posterior lumbar roots. 60% optimal and 10% good results without permanent complications. PMID- 2965479 TI - Failed lumbar disk syndrome. AB - The Authors report 8 cases of Failed Lumbar Disk Syndrome (FLDS), on 210 patients operated for slipped disk. Risk factors and the concept that an abnormality involving either neurons in the substantia gelatinosa or internucial fibers in Lissauer's tract is responsible for pain in patients with FLDS are discussed. Since iatrogenic deafferentation pain response to almost all current forms of therapy is generally poor, the most rational approach remains prevention. PMID- 2965481 TI - [The structure, morbidity level and the frequency of acute diarrheal diseases in children in the preschool period (longitudinal cohort research)]. AB - Some new aspects of acute intestinal infections incidence rate have been revealed in longitudinal cohort study, whose results are proposed for use in the system of prophylactic measures. The group of susceptible children frequently suffering from acute intestinal infections is considered to be of particular scientific and practical importance, for it is this group which determines the level of morbidity in these infections. PMID- 2965482 TI - Topical substances which prevent poison ivy allergic contact dermatitis. PMID- 2965483 TI - The adverse reactions of vehicles and externally applied drugs. PMID- 2965486 TI - Contact urticaria from rubber gloves: a detailed description of four cases. AB - Four cases of contact urticaria from rubber gloves are presented in detail. Three of the four patients, including two atopics had present or previous hand dermatitis. The causative gloves were made of natural rubber. Interestingly, one patient also reacted to her synthetic nitrile rubber gloves which the distributor had sold her as plastic gloves, and explanation for this case is discussed. Contact urticaria should be taken into consideration when there is recalcitrant hand dermatitis in spite of regular use of polymeric protective gloves. Manufacturers should give more accurate and reliable information on the composition of the glove materials to distributors and consumers. PMID- 2965484 TI - Hair bulb accumulation of Langerhans cells in allergic patch tests. AB - The occurrence of occupational allergic contact dermatitis due to 2,3-epoxypropyl trimethyl ammonium chloride (EPTMAC) is reported and supplemented with immunohistochemical and electron microscopic observations. Four young workers developed hand dermatitis at a factory in which modified, cationic starch is manufactured. EPTMAC, a quaternary ammonium compound used as a cationizing chemical in the process, produced allergic reactions in all four patients in epicutaneous testing. The patients had only been in contact with EPTMAC for a short time (one to three months) before developing allergic eczema, which indicates that EPTMAC is a strong sensitizer. Immunohistochemistry showed that dendritic OKT6+ cells (Langerhans cells) increase in the hair follicles and the peribulbar infiltrate during the allergic patch test indicating that hair follicles might actively be involved in delayed type allergic reactions, possibly as a shunt way for allergens. Using electron microscopy, mitotic immunocompetent cells were found in the epidermis during the allergic patch test. PMID- 2965485 TI - Occupational contact dermatitis and contact urticaria caused by epoxy resins. AB - Over a ten-year period (1974-1983) 542 cases of allergic occupational contact dermatitis were diagnosed in our clinic. Epoxy resins caused 71 cases (13.1%) of occupational allergic contact dermatitis (CD), and three cases (0.6%) of irritant CD. One contact urticaria was caused by an epoxy resin hardener, methyl hexahydrophthalic anhydride. Epicutaneous testing with standard epoxy resin 1% in pet. (mainly oligomer of molecular weight 340) produced allergic reactions in all the patients with allergic CD with the exception of three. These three patients had allergic reactions only to their own resins. Testing the 71 patients of allergic CD with epoxy hardeners produced allergic reactions in 19 (26.8%). None of the patients had separate delayed contact allergy to epoxy hardeners without simultaneous contact allergy to standard epoxy resin. PMID- 2965487 TI - [An immunoblastic lymphadenopathy consisting of helper/inducer T lymphocytes, showing monoclonal rearrangement of T cell receptor beta gene]. PMID- 2965488 TI - Forehead skin blood flow in normal neonates during active and quiet sleep, measured with a diode laser Doppler instrument. AB - Changes in forehead skin blood flow during active and quiet sleep were determined in 16 healthy neonates using a recently developed semi-conductor laser Doppler flow meter with-out light conducting fibres. Measurements were carried out at a postnatal age varying from 5 hours to 7 days. The two sleep states could be distinguished in 17 recordings. The mean skin blood flow values during active sleep were significantly higher (p less than 0.01) than those during quiet sleep, the mean increase being 28.1%. The variability of the flow signal, expressed as the coefficient of variation, changed significantly from 23.1% during active sleep to 18.2% during quiet sleep. PMID- 2965489 TI - Cord blood atrial natriuretic peptide (ANP) concentrations--lack of influence of labour stress. PMID- 2965490 TI - Social adjustment and functioning of chronically ill and impaired children and adolescents. AB - A cluster sample of 532 children aged 8 to 19 years with certain impairments were compared to a matched sample of 316 healthy children of the same age. Almost half (46%) of the impaired had a disability, which was mostly in the domain of body disposition ability (in 88%), behaviour (63%) and situational functioning (54%). In the controls, where 12% had some kind of disability, the corresponding figures were 13%, 11% and 15%. The most disabling conditions were CP (67% handicapped), MBD (52%) and epilepsy (42%). The most frequent non-compensated handicaps were those of occupation (44% of impaired), mobility (31%) and social integration (22%). More than one handicap was associated most frequently with CP (76%) and with epilepsy (36%). Neurological impairments are more disabling and handicapping than non-neurological ones, with the exception of mobility and occupation handicaps in asthmatics. PMID- 2965491 TI - [Regulation of cardionatrin on the beating rate of cultured myocardial cells of neonatal rats]. PMID- 2965492 TI - A decade with percutaneous transluminal coronary angioplasty. PMID- 2965493 TI - A new oral contraceptive, Femovan. PMID- 2965494 TI - Plastic surgery for the management of palmoplantar keratodermia (palmoplantoneoplasty). AB - Keratodermia is an incurable genetic and regional disease located in the palmar and plantar regions. The author reports his experience with five cases of palmoplantar keratodermia that were treated by grafting onto the soles and the palms skin taken from the calves and the thighs. PMID- 2965495 TI - Antidromic vasodilatation and neurogenic inflammation. AB - Antidromic stimulation of the lumbosacral dorsal roots of the rat 1.) evokes a long-lasting increase in cutaneous microcirculation of the paw as detected by the laser Doppler flowmeter, and 2.) induces plasma extravasation in the innervated skin areas and various pelvic organs. Both responses are strongly inhibited or abolished by systemic or local capsaicin desensitization. Cutaneous vasodilatation is evoked already by 1-2 pulses. Desensitization of the volar skin of the forearm abolishes the flare reaction both in the "afferent" and "efferent" side of the axon reflex. A dual sensory-efferent function for capsaicin-sensitive nociceptors is suggested. This local neuroregulatory mechanism mediates neurogenic inflammation, cutaneous vasodilatation and flare reaction not only when the receptors are activated by antidromic stimuli, but also when their orthodromic excitation by chemical means occurs. PMID- 2965497 TI - [The local blood flow of the rabbit bladder measured by electrochemically generated hydrogen gas]. AB - The regional blood flow of the urinary bladder of rabbits was measured by electrochemically generated hydrogen gas. Then the effects of increased bladder capacity, and of systemic administration of scopolamine and neostigmine bromide on regional blood flow were examined. The local blood flow of the rabbit bladder was 75.9 +/- 8.4 ml/100 g/min (mean +/- S.E.) and it decreased by increased bladder capacity. It was also decreased by scopolamine bromide, while it was increased by neostigmine bromide. PMID- 2965496 TI - Flare up of joint inflammations induced by cloned helper T cells. Retention of the helper T cells. AB - Joint inflammations were induced in mice by intra articular (ia) injection of cloned helper T cells specific for methylated bovine serum albumin (mBSA) together with mBSA. Local injection of mBSA several weeks after waning of a joint inflammation induced by cloned helper T cells caused a flare up reaction. This indicates that the helper T cells persisted in the joint after the primary inflammation. In the present paper we show that the helper T cells can also persist for some time in a knee joint in the absence of the specific antigen and/or an inflammatory reaction. PMID- 2965498 TI - Cinevideodensitometric analysis of the effect of coronary angioplasty on coronary stenotic dimensions. AB - The accuracy and reproducibility of caliper and cinevideodensitometric measurements of coronary stenotic dimensions were compared in radiographic phantom models and in coronary arteriograms of 28 patients undergoing coronary angioplasty. Projected, single-plane coronary cine frames were analyzed by a computer-assisted videodensitometric method, which measures stenotic cross sectional area without assumptions about lesion geometry. The accuracy (2.4%) and precision (+/- 1.9%) of cinevideodensitometry for measuring percent area stenosis in Plexiglas models of eccentric stenotic lesions was superior to the accuracy (24.7%) and precision (+/- 5.4%) of caliper measurements. Interobserver variability was significantly (p less than 0.05) better for cinevideodensitometric (r = 0.98; SEE = 6.4%) than for caliper measurements (r = 0.87; SEE = 13.1%). After angioplasty, percent diameter stenosis measured by calipers fell from 70 +/- 12% to 30 +/- 15%. Mean percent area reduction measured by cinevideodensitometry fell from 89.1 +/- 8% to 40.1 +/- 22% and stenotic area increased five-fold, from 0.59 +/- 0.5 to 3.47 +/- 1.6 mm2. Pre and post PTCA gradients did not correlate with lesion dimensions. Cinevideodensitometric measurements of absolute stenotic dimensions were more reproducible than relative measurements expressed as a percentage, due to the tapered caliber of normal arterial segments. Thus, cinevideodensitometric measurements were more accurate and reproducible than caliper measurements. The angiographic effects of coronary angioplasty are best measured by cinevideodensitometry, because residual lesions post PTCA are often eccentric, have indistinct margins, and are better characterized by changes in area than by changes in diameter. PMID- 2965499 TI - Influence of exercise on plasma atrial natriuretic factor levels in patients with myocardial infarction. AB - The influence of dynamic exercise on plasma atrial natriuretic factor (ANF) levels was studied in a group of 10 patients with myocardial infarction (MI) and five patients with atypical chest pain (control group). Exercise protocol consisted of three fixed workloads (25, 50, and 75 watts) every 4 minutes with the use of a supine bicycle ergometer. Plasma ANF levels and hemodynamic indices were measured before, during, and 10 minutes after exercise. In the MI group, plasma ANF levels significantly increased at the 75-watt workload and significantly decreased at 10 minutes after exercise, whereas in the control group, the increase in plasma ANP levels after a 75-watt workload, compared with those at rest, was not significant. Significant correlations of pulmonary artery wedge pressure, right atrial pressure, mean arterial pressure, and heart rate to plasma ANF levels were observed at four points obtained before and during each stage of exercise in the MI group. Furthermore, a significant correlation between maximal creatine kinase levels and plasma ANF levels at a 75-watt workload and a significant inverse correlation between left ventricular ejection fraction and plasma ANF levels at a 75-watt workload were observed. These results suggest that the increase in the circulating ANF level during exercise in MI is associated with elevated atrial pressure resulting from left ventricular dysfunction and that measurement of ANF during exercise may be an indication of the severity of MI and associated left ventricular dysfunction. PMID- 2965500 TI - Current concepts in unstable myocardial ischemia. PMID- 2965501 TI - Coronary angioplasty of multiple vessels in corrected transposition with situs inversus. PMID- 2965502 TI - Determinants and protective potential of coronary arterial collaterals as assessed by an angioplasty model. AB - Two indexes of collateral blood flow, the ratio of distal coronary occlusion pressure/aortic pressure (DCOP/Pao) and angiographic collateral class were determined during elective angioplasty in 36 patients with normal left ventricular function. The association between collateral indexes and 8 anatomic and clinical variables was assessed. A reduction in luminal diameter by greater than or equal to 70% predicted angiographically demonstrable collaterals with 100% specificity and 85% sensitivity. Lesion severity (stenosis) correlated with both collateral class and DCOP/Pao: DCOP/Pao = 2.8809 - 0.0729 X stenosis + 0.00049 X stenosis. The data suggest a quantitative relation between lesion severity and collateral development beyond a threshold value of 70% stenosis. Left ventricular ejection fraction during ischemia caused by balloon occlusion (EFo) was found to be primarily determined by lesion location; however, collateral flow modified EFo significantly. For mid-left anterior descending and right coronary artery: EFo = 59 + 26 X (DCOP/Pao); for proximal left anterior descending artery: EFo = 24 + 89 X (DCOP/Pao). A model predicting the hemodynamic and clinical consequences of abrupt coronary closure based on lesion location and severity was developed. In the second study phase, this model was tested retrospectively in a different group of 23 patients who experienced coronary occlusion as a complication of angioplasty. The data of both study phases suggest that left ventricular function and clinical outcome after abrupt coronary closure are determined by an interaction between location of the coronary artery obstruction and the amount of collateral flow. Lesion severity and the extent of functional impairment resulting from abrupt coronary closure are inversely related. PMID- 2965503 TI - Helicopter transport of patients during acute myocardial infarction. AB - Initial experience with a regional system of emergency helicopter transport of patients with acute myocardial infarction (AMI) referred for emergent cardiac catheterization and percutaneous transluminal coronary angioplasty (PTCA) is described. Two hundred fifty patients with AMI were transported from within a 150 mile radius to Duke University Medical Center over a 15-month period. All patients were within 12 hours of onset of symptoms. Thrombolytic therapy was administered to 240 (96%) patients (72% before or in-flight). The time to administration of thrombolytic therapy ranged from 30 to 120 minutes (median 180), while the time to arrival in the interventional catheterization laboratory ranged from 105 to 815 minutes (median 300). The flight time was 12 to 77 minutes (median 31). Most patients had 1- or 2-vessel coronary artery disease; the baseline ejection fraction ranged from 27 to 70% (median 42). Transient hypotension was the most common complication both pre-flight and in-flight. Third degree atrioventricular block and nonsustained ventricular tachycardia were the next most common complications. Ventricular fibrillation or sustained ventricular tachycardia occurred before takeoff in 38 patients (15%). No patients had ventricular fibrillation, asystole or respiratory arrest during transport. Fluid boluses for hypotension were the most common intervention. Five patients required cardiopulmonary resuscitation in-flight; 3 before lift-off and 2 required a brief period of cardiopulmonary resuscitation during sustained ventricular tachycardia. Fourteen patients had pressor therapy, military antishock trousers or both to maintain adequate blood pressure. Neither cardioversion, defibrillation nor intubation were performed in-flight. Thus, inflight complications are infrequent and can be managed en route to an intervention center.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965505 TI - Surgical and medical results of complete atrioventricular canal. PMID- 2965507 TI - Early results of balloon angioplasty of native aortic coarctation in young adults. PMID- 2965506 TI - Treatment of recurrent restenosis with repeat percutaneous transluminal coronary angioplasty. PMID- 2965504 TI - Comparison of two dose regimens of intravenous tissue plasminogen activator for acute myocardial infarction. AB - Two dosing schedules of intravenous tissue plasminogen activator (t-PA) for acute myocardial infarction were compared in a multicenter trial. At 2.95 +/- 1.1 hours from onset of chest pain, 386 patients received 150 mg of intravenous t-PA. For the first 178 patients (group A), 60 mg were given in the first-hour dose and the remaining 90 mg were infused over 7 hours. In the subsequent 208 patients (group B), the first-hour dose was 1.0 mg/kg and the remaining 150 mg were given over 5 hours. At initial angiography 94 +/- 30 minutes into therapy, the infarct vessel patency was 64% in group A versus 75% in group B (p = 0.02). By final angiography with up to 4 selective contrast injections, patency was 68% versus 77%, respectively (p = 0.06). Repeat angiography at 7 to 10 days demonstrated reocclusion in 17% of group A and 13% of group B patients (p = 0.35). There was no difference in fibrinogen nadir or mean hematocrit drop between the 2 groups: 120 mg/dl and 11 points, respectively, in group A compared with 120 mg/dl and 10 points in group B. However, bleeding was reduced in group B patients as evident by a decrease in requirement for greater than or equal to 2 units of packed red blood cell transfusion (group A 36%, group B 27%, p = 0.05) and lower incidence of gastrointestinal bleeding (group A 12%, group B 4%, p = 0.002). To further study the importance of weight adjustment, patients were divided into 2 groups according to weight (less than or equal to 90 kg versus greater than 90 kg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965508 TI - Surgical removal of the intracoronary portion of a fractured angioplasty guidewire. PMID- 2965509 TI - Practice preservation plan. PMID- 2965511 TI - Long-term evaluation of the hepatitis B vaccine (Heptavax-B) in hemodialysis patients. AB - Hemodialysis patients were screened for hepatitis B surface antibody (anti-HBs) prior to immunization at two teaching hospitals. Thirty-one of 111 patients (28%) had baseline sera positive for anti-HBs, while anti-HBs was found in 30 of 420 (7.1%) health care employees (P less than 0.001). A total of 72 hemodialysis patients (mean age, 55.7), received the hepatitis B vaccine (Heptavax-B, Merck Sharp & Dohme, West Point, PA). The responder rates (34 of 72; 47%) and nonresponder (38 of 72; 53%) rates were similar to previous reports. Neither age (P greater than 0.05) nor injection site (P greater than 0.05) appeared to influence results. Nonresponders (16 of 17; 94%) who were given a fourth vaccine dose also failed to mount an antibody response. Of the 34 responders, 18 were followed by serial anti-HBs determinations. Seven transient responders (7 of 18; 39%) were identified, and anti-HBs fell below 10 S/N (sample/control counts per minute) within 12 to 15 months of the first vaccine dose. A fourth dose was administered to this group and it extended the presence of serum anti-HBs (S/N greater than or equal to 10) in four of six patients for another 2, 8, 10, and 15 months, respectively. Antibody persisted but declined over the study period in the remainder of responders followed serially (11 of 18; 61%). When compared with those responders who lost anti-HBs, those with persistent antibody had higher anti-HBs values at 7 (P less than 0.02) and 12 months (P less than 0.005) after the first injection, and were younger (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965510 TI - Seroresponse to hepatitis B vaccine in patients and staff of renal dialysis centers, Wisconsin. AB - Since June 1982, the Wisconsin Chronic Renal Disease Program has provided reimbursement for hepatitis B virus vaccine and pre- and postvaccine serologic testing for patients and staff in all Wisconsin renal dialysis centers. Between July 1, 1982 and June 30, 1984, 444 dialysis patients in 22 renal dialysis centers and 128 staff from nine renal dialysis centers each received three doses of hepatitis B virus vaccine and were tested more than 14 days after the third dose. Antibody response (antibody to hepatitis B surface antigen) occurred in 208 (47%) patients and was age-dependent; the rate of antibody response ranged from 86% (for those aged less than 30 years) to 32% (for those aged over 70 years). Among staff, the rate of antibody response ranged from 96% (for those aged less than 35 years) to 71% (for those aged over 45 years). Results of a multiple logistic regression analysis showed that within age groups, antibody response in dialysis patients was positively associated with length of time on dialysis prior to receipt of vaccine (p = 0.014) and negatively associated with the time interval between the third dose and postvaccine testing (p = 0.0003). Variables that did not enter the regression equation were dialysis mode and frequency, transplant status, concurrent medication use, and injection site. Given the low rates of post-hepatitis B virus vaccine antibody response observed in renal dialysis center patients reported here, the authors recommend that hepatitis B virus vaccine plus alternative hepatitis B virus preventive methods be used. Consideration should be given to a schedule for testing of all vaccinated patients and staff aged over 40 years to ascertain postvaccine susceptibility to hepatitis B virus infection. PMID- 2965512 TI - Surveillance of occupational skin disease using the Supplementary Data System. AB - The utility of the Supplementary Data System (SDS) compiled by the Bureau of Labor Statistics (BLS) in conducting surveillance of occupational skin disease was evaluated by examining 14,703 workers' compensation cases reported to the SDS for the year 1981. Combined with state employment denominators obtained from the BLS Employment and Earnings Program, rates of illness (cases of dermatitis/10,000 employed) calculated for eight major industrial divisions varied significantly according to the criteria used for reporting cases. Despite quantitative variations in the rate of skin disease that depended on specific reporting criteria, the relative ranking of the major industrial divisions remained unchanged, with highest rates of skin disease consistently found in three major industry divisions: agriculture, manufacturing, and construction. This ranking of major industry divisions by rate of dermatitis corresponded extremely well with rankings generated from the 1981 Annual Survey (Spearman rank correlation = .98, p less than .01). At the two-digit level of the Standard Industrial Classification, the rankings based on the SDS had a 77% rank correlation with those from the Annual Survey. Two-digit SIC codes identified from the top 10 in both sets of rankings included crop and livestock production from the agricultural division and leather products, food products, rubber and plastic products from the manufacturing division. PMID- 2965514 TI - Laser-Doppler flowmetry for estimating liver blood flow. AB - Whether laser-Doppler flowmetry can be used to monitor liver blood flow was evaluated in a porcine model in which portal venous blood flow was followed indirectly by electromagnetic flowmetry applied to the superior mesenteric artery, and total hepatic venous outflow was measured directly by using an extracorporeal circuit. Hepatic venous outflow at rest was 23.5 +/- 5.7 ml.kg body wt-1.min-1. Occlusion of the hepatic arterial supply reduced hepatic laser Doppler blood flow to 22%, but hepatic venous outflow only to 80%. Portal venous blood flow remained unchanged or increased slightly. Occlusion of the portal vein, on the other hand, decreased hepatic laser-Doppler blood flow values to 37% and hepatic venous outflow to 13%. Increased hepatic venous outflow pressure, caused by a positive end-expiratory pressure or elevation of the draining cannula, reduced flow and caused approximately equal changes in the three variables, as did reduced flow by step-wise bleeding. From these experiments in the pig it is concluded that laser-Doppler flowmetry on the liver surface clearly reflects relative changes of the total liver blood flow, as exemplified in this study, during venous stasis and bleeding. The technique is, however, more sensitive to blood flow changes in the hepatic artery as compared with the portal vein. PMID- 2965513 TI - Advocacy in word as well as deed. PMID- 2965516 TI - Interaction between ANP and amiloride in renal tubular sodium handling in anesthetized rabbits. AB - To examine whether amiloride, an inhibitor of a conductive sodium channel in the distal tubule, modifies a possible tubular action of atrial natriuretic peptide (ANP), alpha-human ANP (0.05 micrograms.kg-1.min-1) was infused intravenously with or without pretreatment of amiloride (5 mg/kg + 0.04 mg.kg-1.min-1) in anesthetized rabbits. No significant changes in arterial pressure and renal blood flow were observed between two groups after ANP infusion. However, increases in filtered sodium load (FNa) and net tubular sodium reabsorption (RNa) induced by ANP were attenuated after the amiloride pretreatment. These attenuations in FNa and RNa could be due to the failure of increase in glomerular filtration rate, although the reason is not clear. Amiloride pretreatment abolished the ANP induced kaliuresis, whereas ANP-induced increases in urinary sodium and fractional sodium excretion were additive after amiloride pretreatment. Thus, ANP elicited a further natriuretic effect even when the amiloride-sensitive component was blocked. This suggests that there is another mechanism by which ANP induces natriuresis, in addition to any ANP effects mediated by interference with sodium conductive channels. PMID- 2965515 TI - Micropuncture study of the effect of ANP on the papillary collecting duct in the rat. AB - Micropuncture collections were obtained from the terminal collecting duct (CD) at base and tip of the renal papilla of the rat. Group 1 was studied before and during infusion with atrial natriuretic peptide (ANP), group 2 was administered the vehicle only, and group 3 received acetazolamide to increase sodium delivery to the base to a similar extent as after ANP. ANP caused a decrease in blood pressure, a slight increase in GFR, natriuresis, and diuresis. Sodium delivery to the collecting duct at the base of the papilla increased. Between base and tip, sodium reabsorption was inhibited. Tubule fluid sodium concentration (TFNa) was increased at the base and remained high at the tip; in contrast TFNa fell between base and tip in control and acetazolamide groups. After acetazolamide, sodium reabsorption in the terminal CD was not inhibited. These results demonstrate that in vivo ANP 1) increases the delivery of sodium to the terminal CD and 2) inhibits sodium reabsorption in the terminal CD. The findings for chloride were similar to those for sodium. ANP also increased delivery of H2O, K, Ca, and Mg to the CD at the papillary base but did not significantly affect their transport by the terminal CD. PMID- 2965517 TI - Effect of rapid sodium load on circulating atrial natriuretic polypeptide. AB - The hypothesis that an increase in plasma sodium concentration (PNa) causes an increase in circulating atrial natriuretic polypeptide (ANP) was examined in conscious dogs. NaCl solution in small volume (0.3 ml/kg body wt) and at high concentration (20%) was injected intravenously within 2 s to rapidly increase PNa. PNa rapidly increased to 5.1 +/- 0.3 meq/l. Urinary excretion of sodium and water increased to 4.1 and 2.5 times the control levels, respectively. Plasma vasopressin level increased to 3.7 times the control level. Plasma ANP level (PANP) did not change significantly. PANP corrected for sodium-induced hemodilution did not change either. On a different day, a double amount of sodium (0.6 ml/kg body wt of 20% NaCl solution) was intravenously injected into the dogs. PNa increased by 7.3 +/- 0.4 meq/l, which was significantly more than the increase after the 0.3 ml/kg injection. PANP with or without correction for hemodilution again did not change. These results indicate that a rapid increase in PNa within the physiological range does not cause elevation of circulating ANP. This suggests that ANP does not contribute to the regulation of plasma sodium concentration. PMID- 2965518 TI - Role of cardiac atria in the human renal response to changing plasma volume. AB - We examined the role of cardiac atria in the renal response to sequential volume expansion and contraction, during and directly following water immersion. In immersed healthy volunteers (group 1, n = 9) atrial diameter, plasma levels of atrial natriuretic peptide (ANP), and natriuresis increased, whereas renal vascular resistance (RVR) and filtration fraction declined. Each parameter changed in an opposite direction postimmersion. An analysis of transglomerular dextran transport suggests that transglomerular hydraulic pressure difference (delta P) changed in parallel with filtration fraction. Baseline atrial diameter, plasma ANP, RVR, and filtration fraction were significantly elevated in nine recipients of denervated cardiac allografts (group 2). Atrial diameter and plasma ANP changed in parallel with group 1 during and after immersion. However, corresponding reciprocal changes in RVR were smaller and filtration fraction remained constant throughout. From transglomerular dextran transport, we compute that delta P increased progressively during and after immersion, suggesting predominant efferent arteriolar tone. The postimmersed state was associated also with enhanced sodium retention despite sixfold higher plasma ANP than in group 1. These findings are consistent with an effect of cardiac denervation to leave unopposed efferent sympathetic nervous traffic to the kidney. They suggest that the latter is an important modulator of the renal response to changing plasma volume in humans. PMID- 2965519 TI - Long-term hemodynamic actions of atrial natriuretic factor (99-126) in conscious sheep. AB - The hemodynamic and metabolic effects of long-term (5 day) infusion of human atrial natriuretic factor (ANF) were examined in conscious chronically instrumented sheep. Infusion of ANF at 20 micrograms/h, a rate below the threshold for an acute natriuretic effect, decreased blood pressure by 9 +/- 1 mmHg on day 5, associated with a fall in calculated total peripheral resistance. On day 1, ANF reduced cardiac output, stroke volume, and blood volume, effects that were associated with an increase in heart rate and calculated total peripheral resistance and a small decrease in blood pressure. On days 4 and 5 there was a small increase in urine volume and sodium excretion. On day 5 an increase in water intake and body weight was observed. No change was seen in plasma concentrations of renin, arginine vasopressin, glucose, adrenocorticotropic hormone, or protein. This study suggests that the short-term hypotensive effect of ANF results from a reduction in cardiac output associated with a fall in both stroke volume and effective blood volume. However, after 5 days of infusion, ANF lowers blood pressure via a reduction in total peripheral resistance. PMID- 2965520 TI - Immunoreactive atrial natriuretic factor is increased in ovine model of endotoxemia. AB - A bolus of Escherichia coli endotoxin (1.5 micrograms/kg) was administered to chronically instrumented sheep. Immunoreactive atrial natriuretic factor (IR-ANF) was measured in extracted plasma by radioimmunoassay. There was a thirteenfold increase in IR-ANF 2 h after endotoxin administration, and IR-ANF levels remained significantly elevated during the first 6 h. A marked diuresis and natriuresis occurred between 4 and 6 h. ANF not only affects renal function but is also associated with decreased cardiac output, increased peripheral resistance (in sheep), and decreased capillary absorption (in rats). These renal and hemodynamic changes are also characteristic of the early (first 6 h) response to endotoxin. Therefore ANF should be considered as a potential mediator of renal and hemodynamic changes induced by sepsis. It is difficult to determine if ANF elevation is an epiphenomenon or a causative factor, because no antagonist of ANF is currently available. PMID- 2965521 TI - Atrial natriuretic factor attenuates carotid baroreflex-mediated cardioacceleration in humans. AB - We tested the hypothesis that infusions of atrial natriuretic factor (ANF) attenuate carotid baroreflex-mediated tachycardia in humans. The afferent profile from carotid baroreceptors was altered by repeated, intermittent random applications of neck suction (40 mmHg) and neck pressure (20 mmHg) to healthy volunteers during supine control and during infusions of 15 (n = 6) or 25 ng.kg 1.min-1 (n = 9) of alpha-human ANF or during placebo (n = 9). ANF infusions resulted in five- and eightfold increases of plasma ANF, which did not alter base line cardiac interval, blood pressure, respiratory sinus arrhythmia, plasma renin activity, vasopressin, or norepinephrine but did reduce central venous pressure. Reflex bradycardic responses were unaltered during these infusions. Reflex tachycardic responses to carotid compression were significantly blunted (up to 40%) during infusions of ANF and were unchanged during placebo. These responses may be due to a sensitization of cardiac receptors and augmentation of cardiac vagal afferent traffic by ANF, which diminishes reflex cardiac-sympathetic outflow during carotid baroreceptor unloading. PMID- 2965522 TI - Role of atrial pressure and rate in release of atrial natriuretic peptide. AB - To investigate whether atrial natriuretic peptide (ANP) release during paroxysmal tachycardia is due to increased atrial rate or increased atrial pressure, plasma ANP concentrations were measured during atrial pacing at increasing rates in six alpha-chloralose-anesthetized dogs whose atrial pressures were maintained artificially low by balloon occlusion of the inferior vena cava (IVC). These ANP concentrations were compared with those seen during identical increasing atrial rates in the same dogs without IVC occlusion. During incremental pacing without IVC occlusion, pulmonary wedge pressure (PWP; mean +/- SE) rose progressively from 5.3 +/- 1.6 at 200 to 20.2 +/- 2.3 mmHg at 350 beats/min (P less than 0.01), and right atrial pressure (RAP) rose progressively from 2.5 +/- 0.9 at 200 to 6.7 +/- 2.1 mmHg at 350 beats/min (P less than 0.05). At the same time, arterial and coronary sinus ANP concentrations rose from 116 +/- 55 and 339 +/- 91 to 1,126 +/ 226 and 1,960 +/- 456 pmol/l, respectively (P less than 0.01). In contrast, incremental pacing with IVC occlusion produced no significant increase in PWP and RAP. Arterial and coronary sinus ANP concentrations during IVC occlusion were, respectively, 208 +/- 126 and 388 +/- 159 at 200 and 261 +/- 83 and 345 +/- 80 pmol/l at 350 beats/min (NS). This study demonstrates that the release of ANP during tachycardia is primarily dependent on increased atrial pressure and not atrial rate. PMID- 2965523 TI - Chronic atrial appendectomy alters sodium excretion in conscious monkeys. AB - The purpose of this study is to determine whether chronic removal of atrial appendages alters renal response to volume expansion in the conscious monkey. Chronic bilateral atrial appendectomy (ATX) was performed in six animals. Six additional animals served as sham-operated controls. Monkeys were studied 1-2 wk after chronic surgery. The protocol consisted of three consecutive 10-min urine collections followed by 20% ischemic blood volume expansion (VE) and 120 min of post-VE measurements. In sham animals, VE caused an increase in plasma atrial natriuretic peptide (ANP) levels (48 +/- 7 pg/ml to a peak of 108 +/- 34 pg/ml). Urine flow increased from 0.43 +/- 0.07 to 1.07 +/- 0.24 ml/min, sodium excretion increased from 17.9 +/- 2.6 to 74.9 +/- 12.0 mu eq/min, and fractional sodium excretion increased from 0.67 +/- 0.10 to 2.43 +/- 0.28%. ATX attenuated the increase in ANP (34 +/- 8 pg/ml to a peak of 38 +/- 9 pg/ml) in four of six animals. In these animals, renal response to VE was significantly attenuated. Urine flow increased from 0.21 +/- 0.05 to 0.30 +/- 0.01 ml/min, sodium excretion increased from 19.3 +/- 6.02 to 37.8 +/- 5.05 mu eq/min, and fractional sodium excretion increased from 0.79 +/- 0.08 to 1.43 +/- 0.17%. Renal response of two ATX animals with normal increases in atrial natriuretic factor was similar to the sham group. Effect of volume expansion on mean arterial pressure, central venous pressure, and renal hemodynamics was not altered by ATX. These findings demonstrate that bilateral atrial appendectomy in the monkey attenuates the increase in ANP and reduces renal response to VE. PMID- 2965524 TI - [Prevention of obstetric hemorrhage at a hematologic center in Leningrad]. PMID- 2965525 TI - Perturbation of epidermal Langerhans cells in basal cell carcinomas. AB - Epidermal Langerhans cells (LC) play a pivotal role in a variety of local immune responses. Using ATPase-stained preparations, we evaluated the density and morphologic features of these cells in the skin of 12 patients with basal cell carcinoma (BCC). In three patients, we compared the density of LC in the tumor to that in the perilesional skin. In these cases, there was no difference in the numbers of LC/mm2 in the two sites. Perturbations of ATPase-positive, dendritic LC were evident in all specimens. These perturbations included various degrees of disruption of the usually uniform LC network and alterations in the morphology of LC. Many LC had rounded, deformed cell bodies, dendrites that were shortened or completely absent, and variations in the intensity of ATPase staining. With the use of an image analyzer, a measure of the deviation from circularity--a "dendricity index" (DI)--was obtained, based on the area and perimeter of the LC. A significantly lower DI, implying a predominance of less dendritic, more rounded LC, was observed in LC at the BCC surface than in LC present in perilesional skin. This finding raises the possibility that BCC may arise in areas where the morphology, or activity, of these immune cells has been altered; alternatively, the perturbations could reflect an effect of the tumor cells on LC morphology. PMID- 2965526 TI - Histological phases of Bactrim-induced fixed drug eruption. The report of one case. AB - A case is described of a patient with Bactrim-induced fixed drug eruption (FDE). Histological studies were performed at 1 day and 5 days after the drug exposure. While the 5-day-old lesion showed changes classically recognized as FDE, the 1 day-old lesion showed changes typical of a hypersensitivity response: diffuse spongiosis, dermal edema and hemorrhage, neutrophilic polymorphonuclear leukocyte abscess formations, and large numbers of eosinophils. This report underscores the dynamic cellular changes that occur in the evolving FDE lesion. PMID- 2965527 TI - A role for IgE in patch test reactions to inhalant allergens in patients with atopic dermatitis. PMID- 2965528 TI - Development and application of in vitro models for the study of human IgE synthesis. PMID- 2965529 TI - Malfunction in a needle valve. PMID- 2965530 TI - Weals after propofol. PMID- 2965531 TI - Innervation of muscle receptors in the cross-reinnervated soleus muscle of the cat. AB - It has recently been reported (Gregory et al., J. Physiol., 331:367-383, 1982) that cutting a muscle nerve and letting it grow back into the muscle or cross uniting the muscle with a foreign nerve results in major disruption of the normal response patterns of muscle spindles and tendon organs. Here we report observations on the structure of muscle receptors in cross-reinnervated and self reinnervated soleus muscles in an attempt to detect abnormalities that might account for their disturbed function. Eight soleus muscles were reinnervated with the extensor digitorum longus nerve for periods up to 449 days and two were self reinnervated. Following the physiological investigation, the muscle was fixed and stained according to the method of Barker and Ip (J. Physiol., 69:73P-74P, 1963). Spindles and tendon organs were teased from the muscle and photographed. In one cross-reinnervated muscle an attempt was made to isolate all receptors. About two thirds of the normal number of spindles and tendon organs were found. Three categories of receptor were identified: normal, abnormal, and those having no visible nerve endings. There appeared to be little difference in degree of abnormality of receptors in self- and cross-reinnervated muscles. Of the 180 spindles, 3% were normal, 43% had no visible endings, and 54% had abnormal endings. Of 80 tendon organs, 38% were normally innervated, 33% were without visible innervation, and 29% had abnormal endings. We conclude that following long-term cross-reinnervation and self-reinnervation of soleus there is extensive disruption of the normal innervation pattern of both spindles and tendon organs which could account for their functional abnormalities. PMID- 2965532 TI - Clinical pharmacology of doxacurium chloride (BW A938U) in children. AB - The neuromuscular effects of doxacurium were studied in 26 children during halothane-nitrous oxide-oxygen anesthesia. Neuromuscular blockade was measured using electromyographic activity of the adductor pollicis muscle after supramaximal stimulation of the ulnar nerve at 2 Hz for 2 seconds at 10-second intervals. To estimate the cumulative dose-response relation, nine patients received incremental doses of doxacurium (2.5-10 micrograms/kg); nine patients received 27.5 micrograms/kg (the estimated ED95); eight patients received 50 micrograms/kg (1.8 X ED95). The ED25, ED50, ED75, and ED95 (estimated from linear regression plots of log dose vs probit of effect) were 11.5, 14.8, 19.0, and 27.3 micrograms/kg, respectively. Clinical duration (T25) was 27.8 +/- 10.3 (mean +/- SD) minutes at 1 X ED95 and 50.6 +/- 15.6 minutes at 1.8 X ED95. Time to recovery of the train-of-four ratio to 0.75 was 63.1 +/- 32.9 minutes at 1 X ED95 and 108.5 +/- 25.7 minutes at 1.8 X ED95. There were no significant changes in heart rate or mean arterial pressure after bolus administration of any dose of doxacurium. PMID- 2965534 TI - [Selection of the components of general anesthesia during diagnostic laparoscopy and translaparoscopic operations for restoring the reproductive function in women]. PMID- 2965533 TI - [Precurarization with antidepolarizing muscle relaxants during induction of anesthesia using fentanyl]. PMID- 2965535 TI - Hemodynamic effects of prostacyclin infusions in healthy volunteers. AB - The selective hemodynamic effects of an intravenous infusion of prostacyclin on skin and on forearm blood flow during increasing doses up to 6 ng.kg.-1 min.-1 were evaluated in 12 healthy volunteers. The highest dose raised total skin blood flow (p = 0.0001) as measured by laser Doppler with a concomitant rise in finger skin temperature (p = 0.02). Capillary perfusion as measured by transcutaneous oxygen monitoring remained relatively unaffected (p = 0.25). Cold-induced vasoconstriction and the recovery thereafter were not influenced. Diastolic blood pressure tended to decrease (p = 0.06), but there was no change in systolic blood pressure, forearm blood flow, and calculated forearm vascular resistance. The authors conclude that prostacyclin may have dissimilar vasoactive potencies in different vascular beds as shown by a pronounced increase in total skin blood flow, owing to vasodilation of shunt vessels, simultaneously with an insignificant change in nutritional skin blood flow and forearm muscle blood flow. PMID- 2965536 TI - Transluminal dilatation of the internal carotid artery in fibromuscular dysplasia -a case history. AB - In a fifty-one-year-old female patient, bilateral fibromuscular dysplasia of the internal carotid artery was treated during surgery with intraluminal dilatation. The backflow excludes the occurrence of cerebral embolism. The method and its possibilities are discussed. PMID- 2965537 TI - Human alveolar macrophage suppression of lymphocyte proliferation. Accessory characteristics for the generation and functional expression of con A-induced suppressor cells. AB - Human alveolar macrophages (AM) provide less accessory support for mitogen- and antigen-stimulated lymphocyte proliferation than do autologous blood-derived macrophages (BM). This is at least in part due to suppression mediated by AM, but the mechanism of such suppression is not understood. To determine whether AM mediated suppression is related to AM interaction with suppressor lymphocytes (Ts), we examined the accessory cell function of both AM and BM for the generation and functional expression of Ts induced by concanavalin A (Con A). The data indicate that human AM are equivalent to BM for the generation of Con A induced Ts, but AM mediate less suppression of Con A-induced Ts, once generated, than do BM. Addition of indomethacin did not increase lymphocyte proliferation when AM served as accessory cells. Supernatant fluids of Con A-exposed AM promoted greater proliferation of human T-lymphocytes and mouse thymocytes than did supernatant fluids from Con A-exposed BM. Interleukin-1 inhibitor activity was not detected in the supernatant fluids. These observations make it unlikely that soluble factors alone account for AM-mediated suppression. Thus, at least for the Con A-induced Ts system, AM do not mediate suppression either via better generation or greater functional expression of suppressor cells relative to BM. The paradoxically greater proliferation of lymphocytes stimulated by the supernatant fluids of Con A-exposed AM raises the possibility that suppression of Con A-stimulated lymphocyte proliferation observed when AM serve as the accessory cells in primary culture may be related to AM secretion of a molecule with IL-1 like activity. PMID- 2965538 TI - [Chondrodysplasia punctata associated with Down's syndrome. Presentation of a case and review of the literature]. AB - A case of chromosomic aberration and chondrodysplasia punctata is presented. Nine similar have been previously reported being this case first described in Spain. Hypothesis arouse from these observations suggest that they are different entities but also permit to include bone alterations among chromosomic aberration signs. PMID- 2965539 TI - [Laparoschisis. Apropos of 6 cases]. AB - We report on 6 laparoschisis treated in the last 5 years. Five patients survived whereas one died at the age of 41 days. We review clinical aspects of this malformation and point out the need for a prompt operation and an adequate nutritional management. PMID- 2965540 TI - [The Spanish Society of Odontopediatrics]. PMID- 2965541 TI - Depression of T cells in Bell's palsy. AB - We analyzed T-lymphocytes and their subsets in 20 patients with Bell's palsy. We used the monoclonal antibodies anti-Leu-1, anti-Leu-2a, and anti-Leu-3a directed against T-lymphocytes and T suppressor/cytotoxic (Ts) and T helper/inducer (Th) cell subsets. Blood samples taken in the acute and convalescent phases of the illness were compared. The percentage of T cells (Leu-1+ cells) was decreased during the acute stage, mainly because of a reduction of the Th cell subset (Leu 3+ cells). No significant alteration was found in the Ts cell subset (Leu-2+ cells). The T cell and Th cell subset depressions were restored to normal within 4 to 6 weeks of the onset of the palsy. The T cell changes in the peripheral blood in the acute stage of Bell's palsy indicate the involvement of cellular immune mechanisms. PMID- 2965542 TI - Acoustic analysis of voices of patients with neurologic disease: rationale and preliminary data. AB - This paper presents a rationale for acoustic analysis of voices of neurologically diseased patients, and reports preliminary data from patients with myotonic dystrophy, Huntington's disease, Parkinson's disease, and amyotrophic lateral sclerosis, as well as from individuals at risk for Huntington's disease. Noninvasive acoustic analysis may be of clinical value to the otolaryngologist, neurologist, and speech pathologist for early and differential diagnosis and for documenting disease progression in these various neurologic disorders. PMID- 2965543 TI - A decade of PPNG in Singapore. AB - It has been more than 10 years since PPNG was first diagnosed in Singapore. Since then the prevalence of PPNG has increased progressively, and is now maintained at between 30 to 35% of all gonococcal infections inspite of a decrease in the overall incidence of gonorrhoea in Singapore. PPNG infections have been recorded with all forms of complicated gonorrhoea including pelvic inflammatory disease, ophthalmia and disseminated gonococcal infection. Penicillin alone cannot be used to treat gonorrhoea. The use of antibiotics active against PPNG and NPPNG both in uncomplicated and complicated gonorrhoea as first-line therapy is now essential. The medical cost of treatment of gonorrhoea has increased by 100 to 200% as a result of the emergence of PPNG. PMID- 2965545 TI - [Association of Hirschsprung disease, Meckel's diverticulum and gallbladder calculi in a young Down's syndrome patient]. AB - Report of a case of Hirschsprung's disease associated with Meckel's diverticulum, gall stones and trisomy 21, in an 18 year old woman, operated for a bowel obstruction due to a volvulus secondary to bowel distention. A brief review of the literature is presented on the association Hirschsprung's disease-trisomy 21. PMID- 2965544 TI - IgE and immune complex glomerulonephritis. AB - A case is presented of immune complex proliferative glomerulonephritis associated with peripheral eosinophilia, renal deposits of IgE, and eosinophils in glomeruli and interstitial tissue. Two sequential renal specimens are described. This study also reviews prior reports of IgE in glomerulonephritis and discusses possible explanations for the presence of IgE. PMID- 2965546 TI - Ganglioside biosynthesis in rat liver: effect of UDP-amino sugars on individual transfer reactions. AB - Several glycosyltransferases participating in ganglioside biosynthesis were measured in Golgi-rich fractions from rat liver. Addition of those UDP-amino sugars to the enzyme assays which accumulate in liver after treatment of rats with D-galactosamine inhibited the transferases to different degrees. The simultaneous presence of UDP-GalN, UDP-GalNAc, UDP-GlcN, and UDP-GlcNAc in concentrations resembling their overall content in livers 6 h after D galactosamine administration led to an inhibition of the glycolipid galactosyltransferases, GL2 and GM1 synthases of 44 and 64%, respectively. GM2 synthase was moderately inhibited whereas the sialyltransferases (GM3, GD3, and GD1a synthases) were almost unimpaired. Induction of liver cell damage by D galactosamine did not cause any change of glycosyltransferase activities as determined in rat liver homogenates and Golgi-rich fractions. These results indicate a possible role for UDP-amino sugars in the depression of ganglioside biosynthesis observed in vivo after GalN administration. PMID- 2965548 TI - Acquired diffuse keratoderma of the palms and soles with bronchial carcinoma: report of a case and review of the literature. PMID- 2965547 TI - [Combination chemotherapy using low-dose epirubicin and vindesine in patients with metastatic breast cancer]. AB - The therapeutic efficacy of a regimen containing epirubicin (EPI) and vindesine (VDS) was evaluated in patients with breast cancer who were given a dose of 20 mg EPI and 3 mg VDS every 2 weeks. Daily doses of 400 mg UFT and 20 mg tamoxifen (1,200 mg MPA) were also administered. Fifteen patients were evaluable for response, and the results were as follows. 1) Four patients obtained a complete response (CR), and 3 a partial response (PR), giving a response rate of 46.7%. 2) The mean duration of response was 8.6 months at the time of writing. 3) Leukopenia (less than 3,000) was encountered in 60%, but this was generally mild and the regimen was well tolerated. These preliminary results showed the efficacy of this regimen, although further studies will be required. PMID- 2965549 TI - Acitretin in the symptomatic therapy for severe recessive x-linked ichthyosis. AB - Eight men with severe recessive x-linked ichthyosis were treated with acitretin, the main metabolite of etretinate, during four months. All of the patients showed marked clinical improvement of scaling during therapy. Hypervitaminosis A-type adverse reactions were observed in all patients. Although the overall tolerance was good, therapy was interrupted in one atopic patient because of pruritus. There were no undesirable laboratory changes in values. Thirty-five milligrams of acitretin daily provided the best efficacy, with minimal side effects. The beneficial effect of this retinoid lasted between four and six weeks after therapy was stopped. These results suggest that acitretin is a useful agent in the symptomatic therapy of severe recessive x-linked ichthyosis resistant to topical therapeutic modalities. Good results with this agent can be achieved with interval therapy adjusted to seasonal variations of the skin symptoms. PMID- 2965550 TI - Papillon-Lefevre syndrome. Ultrastructural study and successful treatment with acitretin. AB - Four siblings affected by Papillon-Lefevre syndrome (PLS) ranged in age from 2 to 11 years. The parents were unaffected and parental consanguinity was present. The 2-year-old girl showed the early manifestations of PLS; that is, slight gingival swelling and erythema occurring simultaneously with minimal scaling of palms and soles. The other siblings, aged 5, 8, and 11 years, showed severe periodontopathy with tooth loss and marked palmoplantar keratoderma with a centripetal extension of the keratoses to the limbs and trunk. These three older siblings were treated with acitretin (Ro 10-1670), the free acid of etretinate, with complete clearing of the skin and healing of gingival pockets. Treatment was given for 16 months; teeth that erupted during therapy were free of periodontopathy and remained firmly anchored to the alveolar bone. In two of the children ultrastructural examination of involved skin was performed before and during acitretin treatment. Before treatment a large number of lipidlike vacuoles were found in corneocytes and in granulocytes; tonofilaments were reduced in number, and keratohyaline granules frequently showed a rectangular or globular shape. During treatment with acitretin these abnormalities diminished markedly. Thus, etretin is effective in treating PLS and, if treatment is started at an early age, should allow patients with PLS to have normal adult dentition. PMID- 2965551 TI - Effect of isotretinoin on serum levels of precursor and peripherally derived androgens in patients with acne. AB - Sebaceous glands are stimulated by androgens and can convert them to more active forms. Isotretinoin, however, has a profound inhibitory effect on sebaceous gland size and function. This study evaluated the effect of isotretinoin on serum levels of precursor and tissue-derived androgens. Twenty-four subjects (15 men and nine women) were treated for 20 weeks with 1 mg/kg/d of isotretinoin. Serum samples were obtained at baseline, 8, 16, and 24 weeks, and assayed for precursor androgens--total testosterone (TT), free testosterone (free T), dehydroepiandrosterone (DHEA), dehydroepiandrosterone-sulfate (DHEA-S); and tissue androgens--dihydrotestosterone (DHT), and its metabolite, 3 alpha androstanediol glucuronide (3 alpha-diol G). Isotretinoin had no meaningful effects on precursor androgens, except for producing an elevation of free T in women. In contrast, isotretinoin produced depressions in the serum levels of DHT and 3 alpha-diol G in women and in 3 alpha-diol G in men. These decreases are believed to be the result, rather than the cause, of a reduction in the size of the sebaceous glands: The magnitude of the observed decreases may represent the amount of tissue-derived androgens that sebaceous glands normally contribute to the circulating pool. PMID- 2965552 TI - Developmental expression of C3 receptor on murine epidermal Langerhans cells during ontogeny. AB - The developmental expression of C3 receptor, an important surface marker of murine epidermal Langerhans cells (LCs), was quantitatively studied using an immunohistochemical technique on epidermal sheets and then compared with developmental expression of Ia antigen and membrane ATPase. Anti-Mac-1 monoclonal antibody associated with CR3 was used for detecting C3 receptor and proved positive for LCs by immunoelectron microscopy. Mac-1 positive (Mac-1+) cells showed quite a different distribution from those of ATPase+ and Ia+ cells. Almost the same number of Mac-1+ and ATPase+ cells were present during the embryonic period. The number of Mac-1+ cells gradually decreased from day 1 to day 5 of postnatal life, after which they increased again. Using the double-labeling technique on epidermal sheets at day 1 of postnatal life, it was shown that Ia+ cells possessed membrane ATPase activity and some Mac-1+ cells expressed Ia antigen. On days 4 and 7 of postnatal life all Mac-1+ cells expressed Ia antigen. These findings suggest that Mac-1 antigen observed during the embryonic period gradually fades after birth and is re-expressed after day 5 of postnatal life. PMID- 2965553 TI - A modified photometric quantification of skin surface lipids sampled using the cigarette paper method. AB - In severe cases of acne conglobata the use of substances which considerably reduce the sebum excretion rate, such as 13-cis-retinoic acid, is reasonable. To monitor the effect of such preparation a noninvasive and accurate method for measuring skin surface lipids (SSLs) is required. A new modified photometric quantification of SSLs sampled using the cigarette paper method based on reaction with phosphovanilic reagent was introduced. Our overall results indicate that a 3 h SSLs assessment measured photometrically is as accurate as the gravimetric technique; it is reproducible in situations with low and high levels of SSLs. PMID- 2965554 TI - Association of HLA-B35 with mucocutaneous lesions in Israeli patients with rheumatoid arthritis receiving gold treatment. AB - Seventy-four Israeli patients with rheumatoid arthritis (RA) were studied for possible association between HLA antigens and adverse reactions to gold treatment (aurothioglucose). HLA-B35 was significantly increased in patients who developed gold induced mucocutaneous lesions. These results are in accord with earlier reports, despite the different genetic background of the Israeli RA population and the different type of gold compound used. PMID- 2965555 TI - Failure of oral thiomalate to act as an alternative to intramuscular gold in rheumatoid arthritis. AB - Ten patients with active rheumatoid arthritis (RA) were entered into a pilot study to evaluate the effectiveness of thiomalic acid as a disease modifying agent and to assess its toxicity. Oral thiomalic acid (100 mg) was given daily for up to six months. Changes in disease activity were recorded monthly and all side effects noted. No patient recorded any improvement in subjective well being, pain score, or duration of early morning stiffness. No significant change occurred in articular index or haemoglobin (Hb); the erythrocyte sedimentation rate (ESR) showed a tendency to increase. Only three patients completed six months' treatment; six withdrew because of toxic reactions (three with rashes and three with severe gastrointestinal upset) and one because of lack of effect. Thiomalic acid alone appears to have no significant antirheumatic activity and is associated with an unacceptably high incidence of adverse reactions. PMID- 2965556 TI - Arthritis in Down's syndrome. AB - A 31 year old man with Down's syndrome presented with a 10 year history of an inflammatory polyarthritis resembling juvenile chronic arthritis. This case was similar to those already reported of an arthropathy associated with Down's syndrome but was eventually found to be gout. This emphasised the importance of serum uric acid estimation in patients with Down's syndrome and coexistent arthritis. PMID- 2965557 TI - Monoamines and the mechanisms of action of antitussive drugs in rats. AB - The influence of drugs which modify the concentration of brain monoamines on the size of the 50% antitussive dose (AtD50) of morphine (M), dihydrocodeine (DC) and dextromethorphan (DX) was investigated in male Sprague-Dawley rats. The puncture electrode-induced cough method was used for inducing cough. The AtD50 was calculated by the "up and down" method. All drugs were injected i.p. Concentrations of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) in the whole brain were measured by means of high performance liquid chromatography with electro-chemical detection. The values for the AtD50 of M, DC and DX were 1.22, 1.44, and 6.06 mg/kg, respectively. Reserpine (2.5 mg/kg/day, 2 days) produced depression of more than 80% in levels of NE, DA and 5-HT in the brain. This treatment resulted in a substantial reduction in the antitussive effect of the cough suppressants, as evidenced by an increase in the AtD50 of M, DC and DX. p Chlorophenylalanine (PCPA; 300 mg/kg, 24 hr) specifically produced a reduction of more than 70% in the level of 5-HT in the brain. The PCPA-treated rats also displayed an inhibition of the antitussive effect. The AtD50 in reserpine- and PCPA-treated rats was 2- and 4-fold higher, respectively, than the AtD50 for normal rats. alpha-Methyl-p-tyrosine (300 mg/kg, 5 hr) produced a significant reduction in the levels of NE and DA in the brain, but the antitussive effects of M, DC and DX were not altered. These results suggest that 5-HT in the brain may play an important role in the mechanism of action of antitussive drugs. PMID- 2965558 TI - Characteristics of histamine receptors in the isolated and perfused canine coronary arteries. AB - The vascular responses of isolated and perfused canine epicardial coronary arteries to histamine, 2-pyridylethylamine (2-PEA, a selective histamine H1 receptor agonist) and dimaprit (a selective histamine H2-receptor agonist) were studied. Histamine produced only a vasodilatation at small doses (less than 0.3 microgram) and a biphasic response, i.e., vasoconstriction followed by vasodilatation at large doses. 2-PEA produced only a vasoconstriction at small doses and at large doses a slight vasodilatation following vasoconstriction. Dimaprit usually induced only a vasodilatation. The selective histamine H1 receptor antagonist chlorpheniramine shifted the dose-response curves of histamine and 2-PEA for inducing vasoconstriction to the right. Cimetidine (a selective histamine H2-receptor antagonist) caused dose-dependent shifts of the dose-response curves of histamine and dimaprit for vasodilatations to the right. Chlorpheniramine did not attenuate the vasodilatations by 2-PEA and histamine. The removal of endothelial cells by an intraluminal injection of saponin (1 mg) did not affect the vascular responses to histamine, 2-PEA and dimaprit. These results indicate that there are 2 types of histamine receptors in canine epicardial coronary arteries: (i) a H1-receptor mediating vasoconstriction and (ii) a H2-receptor, which is not endothelium-dependent, mediating vasodilatation. PMID- 2965559 TI - [Intracoronary blood flow velocity during angioplasty. Functional guide and indicator of the success of dilatation]. AB - In order to assess the value of a new Doppler-fitted dilatation catheter, changes in blood flow velocities in the coronary arteries were recorded at various stages of transluminal angioplasty in 15 patients with isolated proximal coronary stenosis. A pulsed Doppler probe (emission frequency 20 MHz, recurrence frequency 62.5 MHz) attached to the distal end of the balloon catheter was used to measure blood flow velocities with a Doppler frequency/velocity relationship of about 3.75 cm/s per KHz. During each angioplasty the balloon was inflated 4 times on average, with successive increases of pressure from 7 to 12 atmospheres. Flow velocities were measured before and after each inflation and also when maximum hyperaemia occurred after temporary occlusion by the inflated balloon. Computer assisted analysis of the coronary stenosis was carried out before and after the procedure. Patient's mean age was 53 years. Angioplasty was performed in the anterior interventricular artery (12 cases), the circumflex artery (2 cases) or the right coronary artery (1 case). Anatomical success was assessed by an increase of minimum luminal section area from 1.3 +/- 0.0 to 3.0 +/- 1.2 mm2. Functional success was assessed by a progressive and significant increase of the velocities measured during the first 3 dilatations. No further increase of velocity was observed between the last 2 inflations. Coronary reserve at the end of angioplasty was 2.1, corresponding to the theoretical value calculated taking into account the residual stenosis and the presence of the catheter in situ.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965560 TI - [Refractory unstable angina. Emergency treatment by coronary artery angioplasty under aortic counterpulsion]. AB - We report our experience of 15 transluminal coronary angioplasties performed in an emergency in 13 patients (mean age 67 +/- 8 years) with refractory unstable angina. The procedures were conducted under diastolic counterpulsation effected by means of an intra-aortic balloon. The angina was of the threatening infarction type on 11 occasions and of the threatening extended infarction on 4 occasions. Electrocardiographic abnormalities were recorded in the anterior (11 cases) or in the inferior (4 cases) territory. Three patients had a very alarming haemodynamic status, with acute pulmonary oedema in two and cardiogenic shock in one. After intra-aortic counterpulsation was installed and the clinical signs were stabilized, coronaro-ventriculography was performed, leading to a decision of immediate angioplasty since age, underlying diseases, myocardial function and diffused lesions made most of the patients unsuitable for surgery. In multiple vessel patients electric and angiographic data were used to locate the tight stenosis (92% in all cases) responsible for the acute coronary ischaemia. The stenosis was found to affect the anterior interventricular artery in 9 cases, the circumflex artery in 2 cases, the right coronary artery in 2 cases and a saphenous shunt on the anterior interventricular artery in 2 cases. Immediate arteriographic and clinical success was obtained in 12 out of 15 cases (80%); there were 3 failures with 2 transmural infarctions. Later on, 2 patients underwent aorto-coronary bypass and 1 died of myocardial failure and pulmonary superinfection 3 weeks after the procedure. Ten of our 13 patients were stabilized and were discharged under medical treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965561 TI - [Economic implications of the various treatment methods in myocardial ischemia]. AB - Modern treatments of myocardial ischaemia are effective. Surgical coronary revascularization involves high immediate expenditures, but these are tempered by a subsequent reduction in medical consumption and costs. Coronary angioplasty is a more comfortable revascularization procedure than bypass surgery which it increasingly tends to supersede. The overall health expenditures involved are about 20% lower than those of surgery. Treatment of incipient infarction has recently been introduced. Its generalization implies a reorganization of medical care and a major increase in expenditures. To what extent this increase will be upset by the benefits of avoided or limited infarction can only be determined by detailed and accurate evaluation. PMID- 2965562 TI - [Demonstration of reversible myocardial ischemia in necrotic akinetic areas using endocoronary ECG during angioplasty]. AB - The presence of ischaemic myocardial tissues in necrotic territories and the usefulness of revascularizing these territories are controversial matters. We have determined the existence of this phenomenon by the per-angioplastic intracoronary ECG method, and we have compared the sensitivities of intracoronary ECG and surface ECG. Intracoronary EVG is achieved by using the mobile teflon coated guide wire of coronary angioplasty as a unipolar epicardial electrode. Being epicardial and localized, the electrode explores a limited area of the myocardium, distal to the artery being dilated and momentarily occluded by the balloon during inflations. The study involved 12 patients (mean age 53.7 years) who presented with the following criteria of admission: transmural myocardial infarction, presence of a Q wave on two leads, akinetic segment at ventriculography and coronary stenosis or occlusion amenable to angioplasty. Patients with collateral circulation between the larger epicardial vessels were excluded. Intracoronary ECG recordings were taken before, during and after inflations. In 9 out of 12 patients the ST segments was elevated by 1.3 mV on average between inflations (S.D. 3.14 mV) and by 4.8 mV (S.D. 3.99 mV) during inflations. These high standard deviations were due to major inter- and intra individual variations of ST. The difference was significant (p less than 0.05) at variance analysis. No variation of ST was observed in 3 patients. Only one of the 12 patients had elevated ST on both surface ECG and intracoronary ECG tracings. Thus, intracoronary ECG is a sensitive method to evaluate myocardial ischaemia during coronary angioplasty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965563 TI - Changes in the helper and suppressor lymphocytes in human peripheral blood following maximal physical exercise. AB - Lymphocyte subpopulations and Con A activated suppressors were measured before, immediately after maximal physical effort on bicycle ergometer and after two-hour recovery. The investigations were performed on healthy male volunteers, trained male and female cyclist, nearly at the same age. The measured parameters of maximal effort were comparable in all investigated groups. The significant increase in T8+ (suppressor cytotoxic) cells and moderate increase in T4+ (helper inducer) cells was observed after maximal physical effort; these changes were accompanied by a significant diminution of a T4+/T8+ ratio. Two-hour recovery resulted in the normalization of the investigated parameters. The functional test based on Con A-induced suppressor activity has revealed the divergence, a decrease in Con A-induced suppressor cell activity was noted after maximal effort. PMID- 2965564 TI - Granulocyte factor and graft-versus-host reaction in rats. I. The inhibitory effect of granulocyte factor on the local graft-versus-host reaction in rats. AB - Recent data have proved that PMNLs regulate the immune responses in vitro and in vivo. PMNL specific granules contain an immunoregulatory factor termed the granulocyte factor (GF). The glycogen-induced PMNL infiltration to peritoneal cavity of rats significantly diminished the local GvHR. GF, injected subcutaneously three times at a day of challenge, one and two days after, significantly diminished local GvHR as well. GF treatment before parental lymphocyte injection had no effect on the local GvHR in rats. Inactivation of GF using anti-GF IgG antibodies, abolished the inhibitory effect of PMNLs in GvHR. PMID- 2965565 TI - Immunoregulatory properties of the proteins present in human milk. AB - The immunoregulatory properties of several proteins, isolated from human milk, were investigated. Secretory component and galactothermin exhibited immunoregulatory activities in the in vivo and in vitro assays, generating helper cells, changing the ARFC levels and the resistance of thymocytes to hydrocortisone (HC). In addition, the immunoregulatory action of the P protein and its four fractions was studied. The bulk of the activity was found in the fraction II and III. The results suggest that the postneonatal development of the mammalian immune system is under the surveillance of various immunoregulatory proteins contained in maternal secretory fluids. PMID- 2965566 TI - Identification of the antigen-binding molecule on helper T cell lines specific for L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT). AB - GAT-specific helper T cell lines were shown to bind GAT, coupled to sepharose or sheep erythrocytes. The binding of T cell lines to GAT was specific, since forming of rosettes with GAT-SRBC was inhibited by pretreatment of T cells with free GAT or anti-idiotypic antibodies. Surface iodination of cells and purification of cell lysates on GAT-sepharose, revealed the presence of a 66 KD molecule and a smaller, 33 KD fragment. The proteins could be eluted specifically from GAT-sepharose with a free GAT, or nonspecifically with SDS. The 66 KD molecule could not be reduced by 5% 2-mercaptoethanol. In addition, it was found that the GAT-binding molecule did not bind to the antibodies against the heterodimer (T-cell receptor). The results suggest that the antigen receptor for GAT on helper T cells and inducer suppressor T cells may be related. The GAT binding molecule may be loosely associated with the heterodimer. PMID- 2965567 TI - Modified WAIS-R for patients with speech and/or hand dysfunction. AB - A feasibility study showed that the research edition of the modified WAIS-R for patients with speech and/or hand dysfunction has promise. Four Verbal Scale subtests and three Performance subtests of the modified WAIS-R were correlated with the standard version. The Verbal IQs of an experimental group (n = 16 severely physically handicapped) and Performance IQs of a control group (n = 32 able-bodied adults) on the two versions were not significantly different. In a comparison study of nonretarded patients with severe cerebral palsy (n = 14) or with spinal cord injury (n = 14), the spinal cord group's mean Verbal IQs and Performance IQs on the modified WAIS-R were significantly higher than those of the severe cerebral palsy group, but the difference was relatively larger for Performance IQs than Verbal IQs. Acquisition of motor dysfunction early in development (severe cerebral palsy) compared to later in development (spinal cord injury) may constrain nonverbal intelligence more than verbal intelligence. PMID- 2965568 TI - [Normalization of ventricular compliance and hypertrophy in arterial hypertension. Double-blind study with 2 combinations of a beta blocker and diuretic]. PMID- 2965569 TI - Depo-provera. A lesson in communication. PMID- 2965570 TI - Medroxyprogesterone acetate as an injectable contraceptive. PMID- 2965571 TI - Effects of Ca2+, Mg2+ and calmodulin on the formation and decomposition of the phosphorylated intermediate of the erythrocyte Ca2+-stimulated ATPase. AB - Formation of the phosphorylated intermediate (ECaP) of the human erythrocyte Ca2+ stimulated ATPase (Ca2+-ATPase) was more rapid and reached steady state sooner at 400 microM-Ca2+ than at 1 microM-Ca2+. Calmodulin increased the apparent rate of ECaP formation at 1 microM-Ca2+, whereas at 400 microM-Ca2+, calmodulin decreased the steady-state level of the ECaP without affecting its apparent rate of formation. Removal of endogenous Mg2+ with trans-1,2-diaminocyclohexane-NNN'N' tetra-acetic acid, which decreased both the velocity and Ca2+-sensitivity of the Ca2+-ATPase, did not alter the Ca2+-sensitivity or the apparent rate of formation of ECaP. ECaP formation at high Ca2+ concentrations was not affected by Mg2+ concentrations as high as 1 mM, and the ECaP could be dephosphorylated by ADP and ATP along either the forward or reverse pathways. The results suggest that high Ca2+ concentrations inhibit Ca2+-ATPase activity by preventing dephosphorylation of the E2P complex, rather than by inhibition of the transformation from E1CaP ('high-Ca2+-affinity' ECaP) to E2CaP ('lower-energy' ECaP). PMID- 2965572 TI - Photodynamic effects of haematoporphyrin derivative on DNA repair in murine L929 fibroblasts. AB - Illumination with red light of murine L929 fibroblasts that had been sensitized with haematoporphyrin derivative caused DNA single-strand breaks after a lag time of about 20 min, as revealed by alkaline elution. The cells appeared not to be capable of recovering from this damage. The photodynamic effect of haematoporphyrin derivative on DNA repair was assessed by monitoring the repair kinetics of DNA damage induced by either X-rays, u.v. light (254 nm) or methyl methanesulphonate treatment subsequent to a non-DNA-damaging photodynamic treatment with haematoporphyrin derivative. On 'post-incubation', the normally rapid repair of X-ray-induced DNA strand breaks did not occur, whereas with u.v. light and methyl methanesulphonate treatment after photodynamic treatment prolonged post-incubation resulted in an increase in the number of strand breaks rather than the normally observed decrease. This clearly shows that, after a photodynamic treatment with haematoporphyrin derivative that itself did not cause strand breaks, excision repair in L929 cells is severely inhibited at a stage beyond the incision step. PMID- 2965573 TI - Intermediate forms of human beta-N-acetylhexosaminidase lack activity towards 4 methylumbelliferyl beta-N-acetylglucosaminide 6-sulphate. AB - 4-Methylumbelliferyl beta-N-acetylglucosaminide 6-sulphate was purified from a mixture containing its unsulphated precursor. The substrate was used to test for the presence of functional alpha-subunits in 'intermediate' forms of human beta-N acetylhexosaminidase in samples of normal and pregnancy serum and in extracts of placenta and lymphocytes from a patient with common acute lymphoblastic leukaemia. Intermediate forms in these samples had no activity towards 4 methylumbelliferyl beta-N-acetylglucosaminide 6-sulphate, indicating that they lack alpha-subunits. PMID- 2965574 TI - Specific binding of insulin-like growth factors 1 and 2 to the type 1 and type 2 receptors respectively. AB - 1. Competitive binding and receptor cross-linking experiments have been used to examine the receptor-ligand interactions between three bovine insulin-like growth factors (IGF) and monolayer cultures of myoblasts and fibroblasts. 2. Labelled IGF-2 bound predominantly to the type 2 receptor with negligible label cross linked to the type 1 receptor, notwithstanding the ability of IGF-2 to compete effectively for the binding of IGF-1 to the type 1 receptor. Approx. 100-fold higher concentrations of IGF-1 or the N-terminal truncated (des-Gly-Pro-Glu) IGF 1 (-3N:IGF-1) were required to produce competition equivalent to IGF-2. 3. All IGF peptides, but especially IGF-1, enhanced the binding of labelled IGF-2 to the type 2 receptor of lung fibroblasts. This unusual effect was probably a consequence of the displacement of labelled IGF-2 otherwise bound to a medium protein, a conclusion supported by the demonstration of a 38 kDa membrane protein cross-linked to labelled IGF-2. 4. Both IGF-1 and -3N:IGF-1 bound only to the type 1 IGF receptor in L6 myoblasts, rat vascular smooth-muscle cells and human lung fibroblasts. The peptides competed for labelled IGF-1 binding with potencies in the order -3N:IGF-1 greater than IGF-1 greater than IGF-2 much greater than insulin. Since the IGF peptides were equipotent in skin fibroblasts, it was proposed that the apparently higher affinity of -3N:IGF-1 for receptors in the other cell types was instead a consequence of a low affinity of this peptide for the competing 38 kDa binding protein. PMID- 2965575 TI - Effects of pH, detergent and salt on aggregation of Chinese-hamster-ovary-cell lysosomal enzymes. AB - Upon detergent or hypo-osmotic lysis of CHO-cell postnuclear supernatants or isolated lysosomes at pH 4.8, the lysosomal enzymes beta-hexosaminidase, beta galactosidase, alpha-fucosidase and cathepsin C were readily pelleted, whereas the exogenous marker, long-term-internalized horseradish peroxidase, was not. Salt or pH elevation greatly decreased lysosomal-enzyme pelletability. The results suggest that, under native conditions, lysosomal hydrolases may be aggregated. Aggregation could promote enzyme retention within the organelle. PMID- 2965576 TI - Regulation of intracellular pH in eukaryotic cells. PMID- 2965577 TI - Tyrosine phosphorylation of the receptor for insulin-like growth factor II is inhibited in plasma membranes from insulin-treated rat adipocytes. AB - Insulin action in intact adipocytes leads to a rapid increase in the concentration of receptors for insulin-like growth factor (IGF) II on the adipocyte cell surface, and to a decrease in the [32P]phosphate content of these receptors on the plasma membrane [Corvera & Czech (1985) Proc. Natl. Acad. Sci. U.S.A. 82. 7314-7318]. It has been previously shown that the receptor for IGF-II can be phosphorylated on tyrosine residues by a kinase activity which is expressed in isolated adipocyte plasma membranes. It is now shown that IGF-II receptor phosphorylation in vitro, in plasma membranes derived from insulin treated cells, is markedly decreased compared with the phosphorylation of the receptor in membranes from control cells. This effect of insulin cannot be attributed to an increase in the activity of phosphotyrosyl phosphatase in the membranes. The tyrosine kinase that catalyses the phosphorylation of IGF-II receptors is associated with a fraction of the plasma membrane which is insoluble in Triton X-100. Removal of the Triton X-100-soluble components of the membrane markedly enhances receptor phosphorylation. Moreover, the expression of the inhibitory effect of insulin requires the presence of one or several Triton X-100 soluble components of the plasma membrane. PMID- 2965578 TI - A monoclonal antibody (PL/IM 430) to human platelet intracellular membranes which inhibits the uptake of Ca2+ without affecting the Ca2+ +Mg2+-ATPase. AB - To probe the structure-function relationships of proteins present in the endoplasmic reticulum-like intracellular membranes of human blood platelets a panel of monoclonal antibodies have been raised, using as immunogen highly purified platelet intracellular membrane vesicles isolated by continuous flow electrophoresis [Menashi, Weintroub & Crawford (1981) J. Biol. Chem. 256, 4095 4101]. Four of these antibodies recognize a single 100 kDa polypeptide in the platelet membrane by immunoblotting. One antibody PL/IM 430 (of IgG1 subclass) inhibited (approximately 70%) the energy-dependent uptake of Ca2+ into the vesicles without affecting the Ca2+ +Mg2+-ATPase activity or the protein phosphorylation previously shown to proceed concomitantly with Ca2+ sequestration [Hack, Croset & Crawford (1986) Biochem. J. 233, 661-668]. The inhibition is independent of ATP concentration over a range 0-2 mM-ATP but shows dose dependency for external [Ca2+] with maximum inhibition of Ca2+ translocation at concentrations of Ca2+ greater than 500 nM. This capacity of the antibody PL/IM 430 functionally to dislocate components of the intracellular membrane Ca2+ pump complex may have value in structural studies. PMID- 2965580 TI - Effect of pH and ionophores on the calcium-pumping ATPase of endoplasmic reticulum microsomes from guinea pig pancreas. AB - The effects of the proton-ionophore FCCP, the electroneutral calcium-proton ionophore A23187, pH and oxalate on calcium uptake and (Ca-Mg)ATPase activity were assayed in microsomal vesicles prepared from guinea pig pancreas. The results can be summarized as follows: 1) The optimum pH for ATP-dependent calcium uptake was 7, whereas the (Ca-Mg)ATPase activity was maximal at pH 7.5-8; 2) after a time lag of oxalate-insensitivity, oxalate stimulated strongly the ATP dependent calcium uptake; 3) FCCP uncoupled the process of calcium uptake and ATP hydrolysis; 4) the effects of FCCP on calcium transport and ATPase activity were quite similar to those induced by the calcium ionophore A23187. These results can be explained by the formation of a proton gradient through membrane vesicles during the calcium pumping process. PMID- 2965579 TI - Two systems in vitro that show insulin-stimulated serine kinase activity towards the insulin receptor. AB - Two systems in vitro are described that show insulin-stimulated phosphorylation of the insulin receptor on serine residues. In the first system, insulin receptor was purified partially from Fao rat hepatoma cells by direct solubilization of the cells in Triton X-100 and chromatography on wheat-germ-agglutinin-agarose. Phosphorylation of these preparations with [gamma-32P]ATP in the presence or absence of insulin resulted in 32P incorporation exclusively into phosphotyrosine residues. Serine kinase activity towards the insulin receptor was reconstituted by adding extracts of Fao cells. Prior exposure of the cells to insulin stimulated serine kinase activity towards the insulin receptor in extracts 7.2 fold. A receptor serine kinase activity enhanced by treatment of cells with cyclic AMP analogues was also retained in the reconstituted system. In the second system, insulin receptor and insulin-sensitive serine kinase activity towards the insulin receptor were co-purified from human placenta. The protocol involved preparation of membranes, before solubilization and chromatography on wheat-germ agglutinin-agarose, by using gentle procedures designed not to disrupt a potentially labile association between the insulin receptor and the serine kinase. Serine kinase activity in these preparations towards the insulin receptor was stimulated up to 10-fold by insulin, and the stoicheiometry of serine phosphorylation was estimated to be approx 0.8 mol/mol of insulin receptor for phosphorylations performed in the presence of insulin. Thus a preparation of insulin receptor is described for the first time that is phosphorylated to high stoicheiometry on serine in an insulin-dependent manner. Conditions that facilitate recovery and assay of serine kinase activity are defined and discussed. These systems provide a basis for characterizing the nature of the insulin-sensitive serine kinase that phosphorylates the insulin receptor, and defining its role in insulin action and control of receptor function. PMID- 2965581 TI - Effect of tetraphenylboron on the calcium-dependent ATPase activity of sarcoplasmic reticulum. AB - The apparent effect of tetraphenylboron on the Ca2+-dependent ATPase activity is rather complex being dependent of the physical state of the membrane. In leaky vesicles, tetraphenylboron decreases the affinity of the enzyme for Ca2+, and high concentrations (100 microM) inhibit the enzyme. These effects are mediated by the membrane structure. In native vesicles, tetraphenylboron induces an apparent stimulation of the maximum hydrolytic activity and a decrease of the Ca2+ gradient formed due to specific Ca2+ releasing activity of the mentioned drug. PMID- 2965582 TI - A monoclonal antibody against N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP): a useful tool for preferential detection of naturally circulating ANP. AB - A monoclonal antibody directed against the N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP), named KY-ANP-II, was produced by fusion of a non producing mouse myeloma cell line, X63-Ag8.653, with spleen cells from a BALB/c mouse immunized with synthetic alpha-human ANP (alpha-hANP) conjugated to bovine thyroglobulin. The obtained antibody showed a high affinity for alpha-hANP with a Ka of 6.6 X 10(10) M-1. With this monoclonal antibody, a specific radioimmunoassay for alpha-hANP was established. The minimal detectable level of alpha-hANP in this radioimmunoassay was 0.8 fmol (2.5 pg)/tube, and IC50 was 8 fmol (25 pg)/tube. The radioimmunoassay recognized alpha-rat ANP on an equimolar basis, whereas there was no detectable cross-reactivity with alpha-ANP [4-28] or alpha-ANP [5-28]. Thus, the monoclonal antibody can detect a hormonal form of ANP, alpha-ANP, but not neuropeptide forms of ANP, alpha-ANP [4-28] and alpha-ANP [5-28]. These results indicate that KY-ANP-II becomes a useful tool for preferential detection of a circulating form of ANP and for investigation of the physiological and pathophysiological significance of ANP as a hormone. PMID- 2965583 TI - Differential coupling of smooth muscle and liver vasopressin (V1) receptors to guanine nucleotide binding proteins. AB - We report the reconstitution of the smooth muscle vasopressin V1 receptor functionally coupled to a pertussis toxin-insensitive guanine nucleotide-binding protein. This V1 receptor was spontaneously coupled to this guanine nucleotide binding protein upon solubilization in the absence of agonist, in contrast to our earlier report on the liver V1 receptor, which required agonist for coupling. The smooth muscle V1 receptor was reconstituted as a high affinity receptor (Kd = 5 nM), with a slow rate of agonist dissociation. Upon the addition of guanosine 5' thiotriphosphate, there was a decrease in receptor affinity (Kd = 30 nM) concomitant with an increase in the rate of ligand dissociation. The ability of the smooth muscle V1 receptor to spontaneously couple to a guanine nucleotide binding protein(s) suggests that in the absence of agonist it exists as a high affinity receptor. The smooth muscle V1 receptor may, therefore, be more sensitive to plasma concentrations of vasopressin than its liver homologue. PMID- 2965584 TI - Identification of thromboxane A2 receptor in cultured vascular endothelial cells of rat aorta. AB - The binding site for [3H]SQ29,548, a potent and selective thromboxane A2 (TXA2) receptor antagonist, was studied in cultured vascular endothelial cells (VEC) of the rat aorta. Specific binding of [3H]SQ29,548 to rat VEC at 24 degrees C was saturable, displaceable and of high affinity. Scatchard analysis of equilibrium binding studies indicated that rat VEC contain a single class of binding sites with a Kd of 2.7 nM. The number of maximum binding sites (25.8 fmol/10(6) cells) for [3H]SQ29,548 on rat VEC was respectively 23 and 3.2 times more than that on rat platelets and rat vascular smooth muscle cells. Four TXA2 receptor antagonists and U46619 completely suppressed [3H]SQ29,548 binding to rat VEC, whereas other prostanoids, such as PGD2, PGF2 alpha, PGE1 and Iloprost, displaced the ligand binding only at considerably higher concentrations. These results suggest that the specific receptor for TXA2 is present in rat vascular endothelial cells. PMID- 2965585 TI - Early abnormal development of calmodulin gene expression and calmodulin-resistant Ca2+-ATPase activity in avian dystrophic muscle. AB - We have reported previously that the pectoralis muscle from three month-old dystrophic chickens with signs of myopathy exhibits increased calmodulin content, elevated calmodulin-specific mRNA (Biochem. Biophys. Res. Commun. 137:507-512, 1986), and reduced sarcoplasmic reticulum (SR) Ca2+-ATPase activity in response to calmodulin exposure in vitro (Clin. Res. 34: 725A, 1986). To determine the early time sequence for development of these abnormalities, we have studied muscle from embryos and post-hatched chickens at various ages. Quantitated by dot blot analysis, there was an approximate two-fold increase in calmodulin-specific mRNA in dystrophic muscle as early as 13 days ex ovo which was maintained throughout development up to three months ex ovo. Similarly, Ca2+-ATPase activity measured in SR membranes from chickens as early as 13 days post-hatch was also found to be resistant to stimulation in vitro by exogenous calmodulin, whereas the enzyme from normal muscle was calmodulin-stimulable. These findings suggest that the genetic lesion expressed in the avian dystrophic animal model involves the loss of normal control of intracellular calcium metabolism early in the maturation of the affected musculature and prior to appearance of disease signs. PMID- 2965586 TI - Keratan-sulphate-containing proteoglycans in human osteochondrophytic spurs of the femoral head. AB - Newly synthesized and endogenous proteoglycan was isolated from human femoral head osteochondrophytic spurs. 35SO4-containing keratan sulphate was measured by its susceptibility to endo-beta-D-galactosidase (keratanase) and comprised 15-17% of the two subpopulations of a proteoglycan monomer fraction (D1) resolved by Sepharose CL-2B chromatography (Kav (I), 0.22; (II), 0.78). The size of the newly synthesized keratan sulphate in these fractions was large (Mr greater than 7,000). The hydroxylamine cleavage product of a proteoglycan aggregate fraction (A1) which eluted in the void volume of a Sepharose CL-2B column was immunoreactive with an anti-keratan sulphate monoclonal antibody, 5-D-4. Unlike the proteoglycan aggregate A1 fraction from bovine nasal cartilage, immunoreactivity against 5-D-4 was also found in chromatographic fractions retarded by Sepharose CL-2B. These results lend additional support to our assertion that the osteophyte extracellular matrix consists of hyaline cartilage type proteoglycan. Stimulation of osteophyte proliferation may be useful as a repair mechanism for resurfacing denuded areas of osteoarthritic femoral heads. PMID- 2965587 TI - Effect of nicotinamide on theophylline seizures in rats. PMID- 2965588 TI - Effects of haematoporphyrin derivatives on rat liver endoplasmic reticulum. PMID- 2965589 TI - Polymorphonuclear leukocytes induce damage to the articular cartilage in acute immunologic arthritis in rabbits. PMID- 2965590 TI - Inhibition by lipoxygenase products of TXA2-like responses of platelets and vascular smooth muscle. 14-Hydroxy from 22:6n-3 is more potent than 12-HETE. AB - Lipoxygenase products, which are formed in great amounts in platelets during their activation, have been prepared from arachidonic acid (20:4n-6), the main polyunsaturated fatty acid (PUFA) esterified in platelet phospholipids, and from two major PUFAs of fish fat, eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids. These compounds have been synthesized using platelet suspension as enzymic source, purified by high performance liquid chromatography, and their structure were checked by gas chromatography-mass spectrometry. Their effects were investigated in vitro upon human platelet aggregation induced by 11,9-epoxy methano-analogue of PGH2 (U-46619) and upon thromboxane A2-induced vasoconstriction of rabbit aorta. All hydroxylated fatty acids inhibited U-46619 induced aggregation in a concentration-dependent fashion. Compounds issued from 22:6n-3 were the most potent inhibitors and their IC50 differed significantly from that of 12-hydroxy-eicosatetraenoic acid (12-HETE). Among them, 14-hydroxy docosahexaenoic acid (14-OH-22:6) was the most effective anti-aggregating molecule (IC50:0.45 microM). 10 microM 12-HETE and 14-OH-22:6 inhibited 60% and 75% of smooth muscle contraction induced by TXA2-like material, respectively. At 1 microM, solely 14-OH-22:6 had an inhibitory effect on adrenaline-, angiotensine or histamine-induced contraction. Since thromboxane receptors in platelets and vascular smooth muscle cells present strong similarities, it is concluded that hydroxylated fatty acids can antagonize prostanoid action probably by interfering with their receptor sites. PMID- 2965591 TI - Determinants of porphyrin fluorescence emission spectra of atheromatous plaques. AB - Plaques were obtained via carotid endarterectomy and exposed to the fluorescent dye hematoporphyrin in vitro. Fluorescence emission spectra were used to characterize sites of dye binding. To provide additional information on the role of environment on fluorescence, model systems were studied involving micellar and premicellar systems. The data indicate two distinct fluorescence signals from plaque-bound porphyrin. These signals were at least 50-fold greater than the fluorescence from uninvolved regions of the vessel wall. Fluorescence emission at 618 nm was observed in all plaques examined. A second fluorescence signal at longer wavelengths was associated with the more complex plaques and could be mimicked by mixtures of porphyrin and detergent at a premicellar concentration. These studies suggest the presence of 2 different porphyrin-binding regions in the plaques examined. The first appears to be a predominantly aqueous site which may represent the interior of a cholesterol-rich micellar structure. The second is associated with 'hard' plaque, i.e., collagen and calcification. PMID- 2965592 TI - Impaired learning of a motor skill in patients with Huntington's disease. AB - The ability of patients with Huntington's disease (HD), patients with dementia of the Alzheimer's type (DAT), and amnesic patients (AMN) to acquire the motor skills underlying a pursuit rotor task was assessed. Differences between groups in initial levels of performance were minimized by adjusting the rotation speed of the disk. The HD and DAT groups were also administered a verbal recognition span test. The results showed that the DAT, AMN, and intact control groups all significantly improved their time on target over six test blocks whereas the HD group was severely impaired in the acquisition of this motor skill. On the verbal recognition span test, the DAT and HD groups were significantly and equally impaired, but the HD group evidenced better immediate and delayed recall than did the DAT group. These results provide further evidence that the basal ganglia are critically involved in the acquisition of motor skills. PMID- 2965593 TI - Ten year follow up of patients with single vessel coronary artery disease that was suitable for percutaneous transluminal coronary angioplasty. AB - The 10 year outcome of patients with single vessel coronary artery disease who underwent coronary angiography more than 10 years before and who would have been potential candidates for percutaneous transluminal coronary angioplasty had it been available then is reported. Long term follow up data were obtained in 96 (91 men, five women; mean age 48 years) of 105 consecutive patients with single vessel coronary artery disease (greater than 70% stenosis), judged suitable for coronary angioplasty. Fifty patients had coronary bypass surgery within six months of catheterisation (surgical group) and 46 were treated medically (medical group). At entry to the study more patients in the surgical group had unstable angina, but fewer had a previous history of myocardial infarction. Ten year survival was 91% and remained excellent in all the subsets analysed. Moreover, the quality of life of these patients was good. Over the 10 year follow up, 16 (36%) of the patients treated medically and 13 (26%) in the surgical group were admitted to hospital because of cardiovascular events (including late coronary surgery in four of the patients treated medically). Lastly, 54/69 (78%) of the patients who were employed before catheterisation resumed work and 29 (42%) were still employed 10 years later. Although these data must be interpreted with care because of the limitations inherent in all retrospective studies, it appears that the long term results of conventional medical or surgical treatment are excellent in patients with single vessel coronary artery disease in whom percutaneous transluminal coronary angioplasty is now an option. PMID- 2965594 TI - Percutaneous transluminal coronary angioplasty of one vessel for refractory unstable angina pectoris: efficacy in single and multivessel disease. AB - Forty patients with unstable angina refractory to medical treatment had one vessel percutaneous transluminal angioplasty to the most stenotic lesion in a major coronary artery. The procedure was successful in 35 patients, and the remaining five patients underwent emergency coronary artery bypass graft surgery. The initial success rate (84%) for the 16 patients with single or the 19 patients with multivessel disease (90%) was similar. At early follow up (average nine days) all patients with successful angioplasty remained symptomatically improved; 10 patients (83%) with single and 10 patients (63%) with multivessel disease had negative treadmill stress tests. Five of six cardiac events occurred within the intermediate (average 11 months) follow up period; two patients had recurrent refractory unstable angina, two had angioplasty for progression of disease in a vessel not previously treated by angioplasty, and one had bypass graft surgery. During late (average 26 months) follow up one patient had a non-fatal myocardial infarction while seven patients (58%) with single vessel disease and nine patients (75%) with multivessel disease had negative stress tests; 29 of 40 patients showed long term improvement. PMID- 2965596 TI - Haemodynamic observations during percutaneous transluminal coronary angioplasty in the presence of synchronised diastolic coronary sinus retroperfusion. PMID- 2965595 TI - Dyspnoea of cardiac origin in 67 year old men: (2). Relation to diastolic left ventricular function and mass. The study of men born in 1913. AB - The relation of cardiac dyspnoea to diastolic left ventricular dysfunction was examined in a sample of 67 year old men from the general population of Gothenburg, Sweden. Forty two men with cardiac dyspnoea and 45 controls were selected from the screened cohort of 644 men. M mode echocardiography, apexcardiography, and phonocardiography were used to evaluate heart sounds, diastolic time intervals, aortic root motion (atrial emptying index); peak rate of change in left ventricular dimension, left atrial and ventricular size; and left ventricular mass. There was a significant relation between dyspnoea grade and left ventricular mass and posterior wall thickness. Dyspnoea grade also correlated significantly with the amplitude of the rapid filling wave and the third heart sound, atrial emptying index and left atrial size, the pulmonary component of the second heart sound, and the dimension of the right ventricle. In mild to moderate dyspnoea fractional shortening was normal, but posterior wall thickness and left atrial dimension were increased. The time from the second heart sound to the O point of the apexcardiogram, adjusted for heart rate, was significantly prolonged in mild to moderate dyspnoea, but not in severe dyspnoea. There was a significant decrease of rate adjusted isovolumic relaxation time, probably secondary to altered loading conditions, in severe dyspnoea, but not in mild to moderate dyspnoea. When the effect of systolic function was excluded multivariate analyses showed that the relation between dyspnoea grade and left atrial dimension persisted. The finding that diastolic abnormalities of the heart contributed to the generation of cardiac dyspnoea may have implications for treatment. PMID- 2965597 TI - Genetic control of sebum excretion and acne--a twin study. AB - Sebum excretion and acne grades were measured in 20 pairs each of identical and non-identical like-sex twins. The identical twins had virtually identical rates of sebum excretion (P greater than 0.05), but they had a significantly different degree of acne severity (P less than 0.01). The non-identical twins had significantly different sebum excretion rates (P less than 0.01) and acne grades (P less than 0.01). These findings suggest that sebum excretion is under genetic control and the development of clinical lesions is modified by environmental factors. PMID- 2965598 TI - Recalcitrant acne vulgaris. Clinical, biochemical and microbiological investigation of patients not responding to antibiotic treatment. AB - A small, but clinically significant proportion of acne patients fail to respond adequately to antibiotic therapy. All non-responding acne patients attending the Leeds General Infirmary between September 1985 and April 1986 (49 out of a total of 610 patients; 8%) were investigated with respect to changes in their acne grade, microbial flora and sebum excretion rate. They were compared with 22 age and sex matched untreated control subjects. It was found that in 65% of non responding patients there was no microbiological abnormality, in 16% there was evidence of Gram-negative folliculitis and 20% carried predominantly antibiotic resistant propionibacteria compared with only 5% of untreated controls. There was a significant association between erythromycin therapy and the isolation of erythromycin resistant propionibacteria (P less than 0.001). A causal link, however, has yet to be established between carriage of antibiotic resistant propionibacteria and failure to respond to antibiotic therapy. Our results show that for most patients with recalcitrant acne a non-microbiological explanation must be sought for the lack of therapeutic success. The mean sebum excretion rate (SER) of the non-responding patients was significantly higher than that of matched untreated acne patients (P less than 0.001). A majority of non-responders (69%) had an SER above the upper 95% confidence limit of the control mean. The SER may affect treatment efficacy by influencing the antibiotic concentration within the pilosebaceous ducts. PMID- 2965599 TI - Heterogeneity of CLL: high CD23 antigen and alpha IFN receptor expression are features of favourable disease and of cell activation. AB - The relationship between alpha-interferon receptor (alpha IFNR) numbers, B-cell antigen expression and clinical stage was determined in 35 cases of typical chronic lymphocytic leukaemia (CLL). alpha IFNR numbers were shown to be low in patients with advanced disease and high in those with a more favourable prognosis. The B-cell activation antigen CD 23 was similarly related to stage, being high in more favourable disease. Also, alpha IFNR expression was directly related to CD 23 positivity, but alpha IFN binding was not inhibited by CD 23 monoclonal antibody. There was no correlation between CD 19, 20, 22 and 24 antigen expression and either alpha IFNR numbers or clinical stage. Since CD23 antigen expression is a feature of B-cell activation, we suggest that high CD23 and alpha IFNR positivity are manifestations of an activated cell phenotype and that cell activation in CLL is a feature of favourable disease. PMID- 2965600 TI - Demonstration of the simultaneous activation of Ca2+-independent and Ca2+ dependent ATPases from rat skeletal muscle microsomes. AB - The activation of the Ca2+-independent (basal) ATPase from rat skeletal muscle microsomes is demonstrated in the presence of enough Ca2+ to provide the simultaneous activation of the (Ca2+ + Mg2+)-ATPase. It was achieved taking advantage of the delayed inorganic phosphate (Pi) release due to the formation of a phosphoenzyme complex during the Ca2+-dependent enzymatic cycle, which is evidenced in fast experiments. The microsomes were immobilized on a filter and perfused at constant flow with an incubation medium which was briefly interrupted with a pulse of appropriate reactants to activate the ATPases, at 2 degrees C. Successive samples were collected after passing through the filter, at approx. 0.1 s intervals. The Pi effluent profile coincides with the pattern of the pulse when it activates only the Ca2+-independent ATPase, it appears delayed when the pulse activates only extra Pi production by the (Ca2+ + Mg2+)-ATPase, and it includes a rapid and a delayed component when both Ca2+-independent and Ca2+ dependent ATPases are activated simultaneously by the pulse. PMID- 2965601 TI - ATP-dependent Ca2+ uptake and Ca2+-dependent protein phosphorylation in basolateral liver plasma membranes. AB - ATP-dependent Ca2+ uptake was measured in vesicles of rat liver cell basolateral plasma membranes. Nucleotide-dependent uptake was specific for ATP and observed at pH 7.0 and 7.4/7.5 but not at pH 8.0. ATP-dependent Ca2+ transport was only observed in the presence of Mg2+. Kinetic analysis of ATP-dependent transport revealed an apparent Km in the submicromolar region. Addition of calmodulin and trifluoperazine had no effect on ATP-dependent uptake. A Ca2+-dependent, phosphorylated intermediate with the apparent molecular weight of 135,000 could be demonstrated in the basolateral plasma membranes. Phosphorylated intermediates with apparent molecular weights of 200,000 and 110,000 were demonstrated in microsomes and appeared to contaminate 'basolateral' membrane protein phosphorylation. The results suggest that a 135,000 molecular weight protein is a Ca2+-ATPase and the enzymatic expression of the liver cell basolateral membrane Ca2+ pump. PMID- 2965602 TI - Neurospora crassa pyruvate dehydrogenase complex: component characterization, catalytic properties and location of translation. AB - We propose a simplified procedure for the purification of the Neurospora crassa pyruvate dehydrogenase complex. The purified complex showed four protein bands with apparent Mr values of 53,400, 52,900, 49,000 and 36,900 upon SDS polyacrylamide gel electrophoresis. Components, E2 and E3, of N. crassa pyruvate dehydrogenase complex were identified, respectively, as polypeptides 49,000 and 53,400. It can be deduced that component E1 is constituted of two subunits with Mr values of 52,900 and 36,900. The Km values towards different substrates and the optimal pH and temperature were determined. The protein kinase activity associated with the core enzyme was present in our most highly purified preparations. It was demonstrated that all the protein components of the complex are synthesized under the control of the nuclear genome. PMID- 2965603 TI - Drug activity "in vitro" and "in vivo" on leucocyte chemotaxis. AB - Drug influences on polymorphonuclear leucocytes (PMN) and particularly on PMN migration was investigated. PMN chemotaxis and random migration were assessed in vitro, after drug incubation in vitro and/or administration in vivo. An inhibiting effect on directed migration induced both by fMLP and C5a was demonstrated with some anti-inflammatory drugs whereas random migration was unaffected. The effect was dose-related and linked to drug chemical structure and to physiopathological state of the cells. Similar results may be obtained after administration in vivo. However some examples show that metabolism can inactivate the pharmacological effect of chemical substances and that the route of administration is of prime importance. The activity of new pathological compounds on PMN migration might need to be studied both in vitro and in vivo. In vitro experiments might be performed as a first step of the study. However only in vivo experiments are really representative of potential therapeutical use. Thus we conclude that both experimental series may be performed to appreciate drug interaction with PMN functions during clinical assays. PMID- 2965604 TI - Flush mounted hot film anemometer measurement of wall shear stress distal to a tri-leaflet valve for Newtonian and non-Newtonian blood analog fluids. AB - Wall shear stress has been measured by flush-mounted hot film anemometry distal to an Ionescu-Shiley tri-leaflet valve under pulsatile flow conditions. Both Newtonian (aqueous glycerol) and non-Newtonian (aqueous polyacrylamide) blood analog fluids were investigated. Significant differences in the axial distribution of wall shear stress between the two fluids are apparent in flows having nearly identical Reynolds numbers. The Newtonian fluid exhibits a (peak) wall shear rate which is maximized near the valve seat (30 mm) and then decays to a fully developed flow value (by 106 mm). In contrast, the shear rate of the non Newtonian fluid at 30 mm is less than half that of the Newtonian fluid and at 106 mm is more than twice that of the Newtonian fluid. It is suggested that non Newtonian rheology influences valve flow patterns either through alterations in valve opening associated with low shear separation zones behind valve leaflets, or because of variations in the rate of jet spreading. More detailed studies are required to clarify the mechanisms. The Newtonian wall shear stresses for this valve are low. The highest value observed anywhere in the aortic chamber was 2.85 N/m2 at a peak Reynolds number of 3694. PMID- 2965605 TI - Ti (WT31)-negative, CD3-positive, large granular lymphocyte leukemia with nonspecific cytotoxicity. AB - A case of WT31-, CD3+ large granular lymphocyte leukemia is reported. On surface marker analysis, the proliferating cells were found to be CD3+4-8-16+ and WT31-. By two-color immunofluorescence staining, CD3+4-8- cells were found to be WT31-, and a small population of WT31+ cells expressed either CD4 or CD8. WT31-, CD3+ cells were also identified in a bulk culture of lymphocytes expanded in vitro. Because WT31 monoclonal antibody (MoAb) reacts with the nonpolymorphic epitope of the disulfide-linked heterodimer of the T cell antigen receptor (Ti), the absence of the WT31-reactive Ti determinant may represent an expression of different CD3 associated polypeptides. The rearrangement of the Ti-beta and Ti-gamma genes but not the immunoglobulin gene was demonstrated, and the single pattern of rearrangement indicated the monoclonal origin of the lymphocytes. When the lymphocytes were assayed for their cytotoxicity against K562, MOLT-4, Daudi, and Raji tumor cell lines, a broad spectrum of cytotoxicity for these tumor cells was observed, and the lymphocytes also exhibited antibody- and lectin-dependent cellular cytotoxicity and lymphokine-activated killer activity. Treatment with anti-CD2 and anti-CD3 MoAbs inhibited their nonspecific cytotoxicity. The anti CD3-mediated inhibition of nonspecific cytotoxicity suggested that an as yet unidentified Ti, present in association with the CD3 molecule on these lymphocytes, serves as a specific receptor for target tumor cell recognition. PMID- 2965606 TI - Mechanism of transient adsorption of fibrinogen from plasma to solid surfaces: role of the contact and fibrinolytic systems. AB - The transient detection of fibrinogen on surfaces has been described (Vroman effect) and high-mol-wt kininogen (HK) has been shown to play a role in this reaction. In this study, we attempted to identify the form of HK responsible for preventing detection of the fibrinogen initially adsorbed from plasma to various artificial surfaces and to determine if other plasma components were involved. We compared 125I-fibrinogen adsorption in the presence of normal plasma to plasma deficient in specific proteins. On all surfaces tested, we found that fibrinogen was displaced from the surface. The extent of displacement was greatly reduced, however, but not eliminated in HK-deficient plasma. Factor XII-deficient plasma also showed reduced fibrinogen displacement. These data indicate that HK can actually displace fibrinogen; however, factor XII, or a factor XII-mediated reaction also appears to be necessary for this displacement to occur. Furthermore, when normal plasma was first subjected to extensive contact activation by dextran sulfate, during which the HK was extensively degraded to components smaller than the light chain (as assessed by Western blotting), we observed greatly reduced displacement of fibrinogen. Extensive contact activation of Factor XI-deficient plasma failed to show low-mol-wt derivatives, however, and displacement of fibrinogen was similar to normal plasma that had not undergone extensive activation. These data indicate that HKa (active cofactor produced during contact activation by factor XIIa or kallikrein) is primarily responsible for displacing fibrinogen, and that HKi (inactive cofactor generated by factor XIa) cannot displace fibrinogen. The fibrinogen from all plasma samples looked similar by Western blot analysis, suggesting that fibrinogenolysis was not a component of the Vroman effect. In addition, experiments performed with plasma prechromatographed on lysine agarose showed that a lysine-agarose adsorbable protein may be minimally involved in fibrinogen desorption and a synergism may exist between HK and that protein. PMID- 2965607 TI - Isolation of an abnormal protein C molecule from the plasma of a patient with thrombotic diathesis. AB - Protein C has been purified from the plasma of a patient with thrombotic diathesis. Both before and after isolation, the protein showed reduced capacity to hydrolyze synthetic substrates and to anticoagulate plasma. Proteolysis with the soluble thrombin-thrombomodulin complex proceeded normally and to completion as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Approximately one-third of the protein is functional, indicating a heterozygous defect. Indirect studies suggest that the abnormal component can bind to protein S and phospholipids. Both forms of activated protein C can also incorporate radiolabeled diisopropylfluorophosphate. PMID- 2965608 TI - The bcl-2 gene is rearranged in many diffuse B-cell lymphomas. AB - Southern blotting was used to detect rearrangement of the bcl-2 gene in 104 cases of non-Hodgkin's lymphoma subclassified by the Working Formulation, 24 cases of B cell chronic lymphocytic leukemia (B-CLL) and 14 cases of T cell malignancy. Earlier workers reported rearrangement of this gene (located on chromosome 18) in a major fraction of follicular lymphomas, lymphomas in which a 14;18 chromosome translocation is frequently observed. In the present study, bcl-2 was rearranged in 30% (11 of 37) of follicular lymphomas and 19% (11 of 58) of diffuse lymphomas of follicle center cell lineage. In 18 of 19 samples studied, the rearranged bcl 2 fragment also hybridized with a probe for the joining region of the immunoglobulin heavy chain gene located on chromosome 14, indicating a 14;18 translocation. In lymphomas not derived from follicle center cells, ie, diffuse lymphomas of small B lymphocytes, B-CLL and T cell neoplasms, the bcl-2 gene was always in germline configuration. The frequent rearrangement of bcl-2 in a variety of B cell lymphomas of diffuse morphology (small cleaved cell, large cell, small noncleaved cell and immunoblastic) is noteworthy. PMID- 2965610 TI - Gilles de la Tourette's syndrome in Down's syndrome. PMID- 2965609 TI - The in vitro effect of pirprofen on the surface ultrastructure and function of human platelets. AB - The in vitro effect of pirprofen (Rengasil), an antiinflammatory agent, on the surface ultrastructure and function of human platelets was examined and compared with that of acetylsalicylic acid (Aspirin, ASA), and diclofenac sodium (Voltaren, DS). Incubation with pirprofen induced formation of long, needle shaped pseudopodia, a phenomenon observed also after incubation of the cells with DS. In contrast with ASA and DS, pirprofen induced a marked increase in platelet protein synthesizing capacity. The drug decreased the platelet aggregation to a degree similar to that of ASA and DS. The release of platelet factors 3 and 4 and the level of beta-thromboglobulin following incubation with the drug remained unaltered. PMID- 2965611 TI - Choosing easy chairs for the disabled. PMID- 2965612 TI - Role of helper T-lymphocytes in rejection of UV-induced murine skin cancers. AB - The purpose of this study was to examine the role of helper T-lymphocytes (Th) in the immunological rejection of UV-induced tumors. Mice repeatedly exposed to UV radiation develop suppressor T-lymphocytes that facilitate the growth of UV induced tumors by interfering with host immunity. These suppressor cells specifically blocked the generation of antitumor Th, suggesting that Th may be important in the immunological rejection of UV-induced tumors. The Th activity generated by a UV-induced tumor that grows progressively in normal mice was compared with that generated by a highly antigenic, regressor clone of the same tumor. The regressing tumor cell line generated a much higher amount of Th activity than the parental, progressor tumor cell line. The amount of Th activity generated by a highly antigenic, UV-induced tumor injected into normal mice, in which it regresses, was compared to the Th activity in UV-irradiated mice, in which the tumor grows progressively. Again, tumor regression was associated with a higher amount of Th activity, and this increased activity was detectable in the environment of the regressing tumor. Lyt-1+ cells containing Th activity mediated the regression of a UV-induced tumor when injected with the tumor cells s.c. into immunosuppressed mice. Lyt-1- cells were cytotoxic to tumor cells in vitro but were unable to cause tumor rejection in vivo. These studies suggest that Th play a central role in the immunological rejection of UV-induced tumors. PMID- 2965614 TI - Effects of cis-diamminedichloroplatinum(II) on rabbit kidney in vivo and on rabbit renal proximal tubule cells in culture. AB - The nephrotoxic potential of cis-diamminedichloroplatinum(II) (CDDP) in rabbits, as well as its effect on cell viability, cellular synthetic activity, and specific enzyme activities in rabbit renal proximal tubule cells, was investigated. Male New Zealand White rabbits were given a single i.v. dose of either 2.5 or 5.0 mg/kg CDDP via the ear vein and sacrificed 5 days later. No drug-induced changes were observed in the kidneys of rabbits given 2.5 mg/kg CDDP. However, histopathological examination of kidneys from rabbits administered 5.0 mg/kg CDDP revealed marked tubular degeneration and necrosis, with the majority of lesions being situated in the outer zone of the cortex. This is in contrast to the effect of CDDP in the kidney of the rat where the necrosis is reported to be predominantly localized to the pars recta of the proximal tubule in the outer stripe of the medulla. The results from the in vitro experiments indicated that the viability of cells after a 6-h exposure to CDDP at concentrations up to 100 microM was greater than 95%. However, a dose-dependent decrease in cell viability was obtained after 24 h exposure with a TD50 (50% viability) of approximately 90 microM. In addition, the results after 24 h exposure to CDDP also indicated that Na+, K+-ATPase, a basolateral membrane marker enzyme, and alkaline phosphatase, a brush-border marker enzyme, were inhibited by 35-40% and 20%, respectively. No effect on succinic dehydrogenase, a mitochondrial marker enzyme, was obtained. Inhibition of all three marker enzymes was minimal at 6 h posttreatment. On the other hand, inhibition of DNA, RNA, and protein syntheses was evident as early as 6 h posttreatment with DNA (48-77%) and RNA (36-77%) syntheses being inhibited to a greater extent than protein synthesis (14-33%). These results demonstrate that inhibition of renal synthetic activity by CDDP, rather than its effect on enzyme activity, precedes the onset of cell lethality and may therefore be an important event in the initiation of CDDP induced nephrotoxicity. PMID- 2965613 TI - Antifolate drug interactions: enhancement of growth inhibition due to the antipurine 5,10-dideazatetrahydrofolic acid by the lipophilic dihydrofolate reductase inhibitors metoprine and trimetrexate. AB - The presence of low concentrations of the lipophilic dihydrofolate reductase inhibitors metoprine or trimetrexate, which cause little inhibition in the growth of cultured hepatoma cells in combination with weakly inhibiting concentrations of 5,10-dideazatetrahydrofolate, exhibit greater activity than would be predicted by the activity of the individual components. Growth inhibition by this inhibitor of glycineaminoribonucleotide transferase alone or in the presence of the reductase inhibitors is prevented by hypoxanthine indicating that the combination of drugs is enhancing the activity of 5,10-dideazatetrahydrofolate against purine biosynthesis. H35 hepatoma cells resistant to methotrexate (100-fold) as a result of a transport defect are 40-fold resistant to 5,10-dideazatetrahydrofolate suggesting that this analogue enters hepatoma cells at least in part by the reduced folate coenzyme-methotrexate transport system. The transport-resistant cells are also susceptible to enhanced inhibition of cell growth by low levels of reductase inhibitors in combination with 5,10-dideazatetrahydrofolate. These results have a corollary in an earlier study showing that the same concentrations of metoprine and trimetrexate could enhance the growth inhibition and cytotoxicity of the folate-based inhibitor of thymidylate synthase, 10-propargyl 5,8-dideazafolic acid (Galivan et al., Cancer Res., 47: 5256-5260, 1987). Combinations of 5,10-dideazatetrahydrofolic acid and 10-propargyl-5,8 dideazafolic acid are less growth inhibitory than that predicted by each of the folate analogues alone. It is possible that the effects of all these combinations are related to distortions in the folate pools caused by the folate analogues being used in combination. Two methods of analysis, one graphical and one mathematical, were used to analyze the drug interactions described in this presentation. The enhancement effect seen with the lipophilic dihydrofolate reductase inhibitors and 5,10-dideazatetrahydrofolate clearly represents a supraadditive or a synergistic drug interaction. In contrast the combination of the folate-based inhibitors of purine (5,10-dideazatetrahydrofolic acid) and thymidylate biosynthesis (N10-propargyl-5,8-dideazafolate) exhibit frank antagonism under certain conditions. PMID- 2965615 TI - Diltiazem prevents hypertrophy progression, preserves systolic function, and normalises myocardial oxygen utilisation in the spontaneously hypertensive rat. AB - The effects of diltiazem, a calcium channel blocker, and methyldopa, an adrenergic blocker, on left ventricular hypertrophy and left ventricular function were assessed in spontaneously hypertensive rats and Wistar-Kyoto controls. Diltiazem (30 mg.kg-1/day), methyldopa (400 mg.kg-1/day), or placebo were given with water for six months. Left ventricular function was studied in 12 month old animals using an isovolumetrically contracting heart preparation by measuring maximum developed pressure and myocardial oxygen consumption. Systolic blood pressure was reduced by both drugs but more so by methyldopa. Despite its lesser antihypertensive effect, diltiazem reduced heart to body weight ratios in the spontaneously hypertensive rat to a similar degree as methyldopa (3.4(0.2) and 3.4(0.1) compared with placebo 3.7(0.2), p less than 0.05). Maximum developed pressure increased with methyldopa and diltiazem compared with placebo (188(11) and 200(11) vs 166(11) mmHg, p less than 0.05). Myocardial oxygen consumption was lower in the spontaneously hypertensive rat receiving placebo than in the controls (22.8(3.2) vs 28.3(3.8) ml.min-1.100 g-1, p less than 0.05) and was significantly increased by diltiazem but not by methyldopa (27.9(0.4) vs 24.5(0.6) ml.min-1.100 g-1, p less than 0.05 and NS respectively vs the spontaneously hypertensive rat receiving placebo). Diltiazem and methyldopa normalised the isomyosin composition in the spontaneously hypertensive rat. Myocardial concentrations of energy related metabolites obtained at maximum developed pressure were not different between spontaneously hypertensive rats receiving placebo and controls. However, both diltiazem and methyldopa treated spontaneously hypertensive rats showed a significant reduction in adenosine triphosphate and phosphocreatine and a rise in inorganic phosphate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965616 TI - Suppression of cytotoxic T-cell generation by natural suppressor cells from mice with GVHD is partially reversed by indomethacin. AB - Natural suppressor cells from the spleens of mice with graft-versus-host disease produced across minor histocompatibility barriers inhibit the in vitro generation of alloreactive cytotoxic T lymphocytes by spleen cells from normal mice. The mechanism of natural suppression was studied; suppression does not require direct cell contact with targets. Exogenous interleukin-2 has no effect in reversing suppression while indomethacin partially reverses the suppression mediated by natural suppressor cells. PMID- 2965617 TI - Suppression of immune cell function in vitro by recombinant human transforming growth factor-beta. AB - The influence of recombinant human transforming growth factor-beta (rHuTGF-beta) on B-cell function and antigen-specific T-cell responses in vitro was investigated. The addition of 0.1 ng/ml of rHuTGF-beta to cultures of peripheral blood mononuclear cells (PBMC) stimulated with tetanus toxoid resulted in a 50% inhibition of the PBMC proliferative response as determined by [3H]thymidine incorporation. Further, rHuTGF-beta at 0.37 ng/ml caused a greater than 50% reduction in the number of immunoglobulin G-secreting cells among PBMC induced by pokeweed mitogen. rHuTGF-beta also inhibited the generation of allospecific cytotoxic T lymphocytes (CTL) in the mixed-lymphocyte reaction but had no effect on the cytolytic function of CTL generated in the absence of exogenous HuTGF beta. The results indicate additional immunoregulatory activities for HuTGF-beta and suggest that this factor may play an important role in the regulation of the antigen-dependent immune response. PMID- 2965618 TI - [Analysis of foci of viral hepatitis B in selected areas. Preliminary report]. PMID- 2965619 TI - [Lyme disease--a new infection from natural foci]. PMID- 2965620 TI - [Determination of total immunoglobulin E using enzyme immunoassay]. PMID- 2965621 TI - [Recombinant vaccinia viruses--the basis of new vaccines]. PMID- 2965622 TI - [Comparison of basic media for the culture of thermophilic strains of the genus Campylobacter]. PMID- 2965623 TI - [Food poisoning of staphylococcal origin in Czechoslovakia 1981-1985]. PMID- 2965624 TI - [Evaluation of the bactericidal effectiveness of disinfection agents using a suspension micromethod]. PMID- 2965626 TI - [Spontaneous hematoma syndrome of the abdominal rectus muscles (Differential diagnosis of acute abdomen and other diseases. Personal cases]. AB - The authors examine the problem of the differential diagnosis of spontaneous hematoma of the abdominal rectus muscles versus "surgical" or "non-surgical" acute abdomen and other conditions. The clinical characteristics and investigative methods used are reviewed with a view to avoiding diagnostic errors. In addition, the authors discuss 6 clinical cases which they have observed personally. PMID- 2965625 TI - Influence of acylcarnitines of different chain length on pure and mixed phospholipid vesicles and on sarcoplasmic reticulum vesicles. AB - Palmitoyl-, myristoyl- and lauroylcarnitine destabilize small unilamellar vesicles of 1,2-dipalmitoyl-n-glycero-3-phosphorylcholine (DPPC) and 1,2 dimyristoyl-n-glycero-3-phosphorylcholine (DMPC) into multilamellar liposomes. Their effect on the bilayer is dependent on the acyl chain length of the acylcarnitine, the ratio of the lengths of the acyl chains of the phospholipid and the acylcarnitine and the molar ratio of the phospholipid to acylcarnitine but not the absolute concentration of the acylcarnitine in the solute. Sarcoplasmic reticulum vesicles are broken down by each of the acylcarnitines at concentrations below their critical micellar concentrations (CMC). These three acylcarnitines stimulate the Mg2+, Ca2+-ATPase activity in SR-vesicles to a certain maximum, after which a net inhibition is observed. The maximum degree of stimulation depends highly on acyl chain length: the shorter the chain length, the more effective. In the same concentration range where the Mg2+, Ca2+-ATPase activity is increased, the net Ca2+-uptake is markedly decreased. PMID- 2965628 TI - Efficacy of low-dose cyproterone acetate compared with minocycline in the treatment of acne vulgaris. PMID- 2965627 TI - Midazolam-sufentanil anaesthesia for phaeochromocytoma resection. AB - This is a case report of a 66-year-old patient to whom a combined infusion of midazolam and sufentanil was administered for phaeochromocytoma resection. With the exception of a drop in blood pressure immediately after tumour removal, significant intraoperative haemodynamic stability was observed. There was no need for the intraoperative administration of hypotensive or anti-arrhythmic drugs, nor was any prolonged postoperative anaesthetic effect noted. In this case, the combination of midazolam and sufentanil with N2O:O2 was successful in maintaining cardiovascular stability until the tumour was removed. The consequent drop in blood pressure responded to fluid infusion, a not uncommon event in phaeochromocytoma surgery. Although a prospective randomized study for resection of phaeochromocytoma showed that the choice of the anaesthetic technique is not a crucial factor in determining the patient outcome, we feel this technique of midazolam sufentanil is a worthy alternative to the use of inhalational anaesthetics. The lack of significant myocardial depressive effect of the two drugs, coupled with its simple administration, makes it a useful technique in the anaesthetic management of phaeochromocytoma resection. PMID- 2965629 TI - Rubber allergy among construction workers in a prefabrication construction factory. PMID- 2965630 TI - An acne naevus. PMID- 2965631 TI - Low dose infusion of atrial natriuretic peptide causes salt and water excretion in normal man. AB - 1. The effects of low dose infusion of atrial natriuretic peptide (ANP) were observed in double-blind, placebo-controlled study in six fluid-loaded volunteers. After baseline observations, hourly increments of 0.4, 2 and 10 pmol min-1 kg-1 were infused with continuous observation of heart rate, blood pressure and cardiac output. Plasma ANP, aldosterone, and catecholamines, and urinary volume and sodium excretion, were estimated at half-hourly intervals. 2. ANP infusion resulted in an increase of 35, 98 and 207% in urinary sodium excretion and of 10, 20 and 71% in urinary volume when compared with placebo. Plasma ANP was markedly elevated above placebo levels only during infusion of 10 pmol of ANP min-1 kg-1. 3. No change in heart rate of blood pressure was noted during the study, but a significant fall in stroke volume index was observed during active treatment. Plasma levels of aldosterone and catecholamines were not significantly different on the 2 treatment days. 4. The potent natriuretic and diuretic effects of this peptide at plasma concentrations not significantly elevated from physiological suggest a hormonal role for ANP in the homoeostasis of salt and water balance. PMID- 2965632 TI - Antihypertensive and renal effects of cilazapril and their reversal by angiotensin in renovascular hypertensive rats. AB - 1. The antihypertensive and renal effects of cilazapril, a new angiotensin converting enzyme inhibitor, were evaluated in both two-kidney, one-clip Goldblatt hypertensive rats (n = 11) and normotensive rats (n = 6). 2. Intravenous infusion of cilazapril (1 mg/kg followed by 25 micrograms min-1 kg-1) caused significant reductions of blood pressure from 163 +/- 3 to 122 +/- 4 mmHg and from 157 +/- 2 to 113 +/- 3 mmHg in two separate groups of hypertensive rats and from 124 +/- 1 to 105 +/- 2 mmHg in normotensive rats. The hypotensive effect in terms of absolute value of percentage change was greater in hypertensive rats than in normotensive rats (41 +/- 6 vs 20 +/- 3 mmHg or 25 +/- 4% vs 16 +/- 2%, respectively). 3. Cilazapril increased glomerular filtration rate, urine flow, and absolute and fractional excretion rates of sodium and potassium in the non clipped kidney of hypertensive rats. In contrast, the clipped kidney exhibited a depressed renal function during cilazapril infusion. 4. In normotensive rats, the hypotensive and enhanced renal function responses to cilazapril were much less than those of the non-clipped kidney of hypertensive rats. 5. Superimposed administration of either angiotensin II or angiotensin III during cilazapril infusion completely reversed the blood pressure and bilateral renal responses of cilazapril in both hypertensive and normotensive rats. 6. These results indicate that cilazapril reduces arterial pressure and enhances renal excretion mainly via inhibition of angiotensin II and angiotensin III formation. PMID- 2965633 TI - Effects of angiotensin II and atrial natriuretic peptide alone and in combination on urinary water and electrolyte excretion in man. AB - 1. Atrial natriuretic peptide (ANP) has previously been shown to inhibit the renin-angiotensin-aldosterone system (RAAS) at several different levels. We have now investigated a further non-endocrine, renal interaction between ANP and the RAAS. 2. The effects of ANP and angiotensin II (ANG II) alone, and in combination, on urinary electrolyte and water excretion were studied in eight normal male subjects undergoing maximal water diuresis. 3. ANP caused a significant increase in urine flow and sodium excretion. ANG II alone was antidiuretic, antinatriuretic and antikaliuretic. When ANP was given against a background infusion of ANG II, urinary flow rate and electrolyte excretion increased from a new lower level to reach a value intermediate between that found with ANG II alone and ANP alone. 4. It is concluded that the renal effects of ANP are modified in the presence of simultaneously elevated levels of ANG II and that net water and electrolyte excretion reflect the sum of the opposing influences of each peptide. While this interplay may be non-specific, it is possible that ANP may exert some of its actions by specifically inhibiting the intrarenal effects of ANG II. PMID- 2965634 TI - Effect of 2,3-diphosphoglycerate concentration on the alkaline fragility of phosphofructokinase-deficient canine erythrocytes. AB - 1. Erythrocytes in whole blood samples from dogs with phosphofructokinase (PFK) deficiency had lower 2,3-diphosphoglycerate (2,3-DPG) concentrations, higher ATP concentrations, and were more alkaline fragile than normal canine erythrocytes. 2. Reticulocytes from a PFK-deficient dog contained nearly three times the ATP concentration of normal canine erythrocytes, and had 2,3-DPG concentrations similar to normal canine erythrocytes. 3. PFK-deficient reticulocytes are not alkaline fragile. 4. The erythrocyte 2,3-DPG concentration in whole blood samples from PFK-deficient dogs was increased to normal by in vitro incubation with dihydroxyacetone, pyruvate and phosphate. This incubation resulted in only a slight increase in ATP concentration. 5. The alkaline fragility of these 2,3-DPG replenished PFK-deficient erythrocytes was normal. 6. Findings in this study indicate that the increased alkaline fragility of canine PFK-deficient erythrocytes is the result of decreased intracellular 2,3-DPG concentration. PMID- 2965635 TI - The distribution of phosphofructokinase isoenzymes in the liver of camel, rat and rabbit. AB - 1. The distribution of phosphofructokinase isoenzymes have been compared among camel, rat and rabbit livers. 2. Only a single phosphofructokinase isoenzyme is present in the camel liver which has shown different physical and regulatory properties from the isoenzymes of rat and rabbit liver. 3. The ammonium sulphate precipitation curves of the camel and rabbit enzymes were monophasic, whereas the rat enzyme was biphasic. 4. Rabbit liver phosphofructokinase was slightly more anodic than the rat enzyme, whereas the camel enzyme was the least anodic as shown by the techniques of DEAE-cellulose chromatography and cellulose acetate electrophoresis. 5. Partially purified camel liver phosphofructokinase showed different regulatory properties from the rabbit and rat isoenzymes as the apparent Km values were 0.58, 0.45 and 0.82 mM respectively. PMID- 2965636 TI - Effects of combined ethinylestradiol and desogestrel on hormone profiles and sex hormone binding globulin in women with polycystic ovarian disease. AB - A longitudinal study to determine the effects of combined ethinylestradiol (EE) and desogestrel (DG) on hormone profiles and levels of sex hormone binding globulin (SHBG) were carried out in 19 Thai women with polycystic ovarian disease (PCOD). Evaluations were performed before and after 3 and 6 cycles of treatment. Regularity of cycles was observed in all patients. Most of them had improvement of androgenic symptoms. Body weight decreased slightly after 6 cycles of treatment. During the treatment course, serum luteinizing hormone (LH), testosterone (T), free T and dehydroepiandrosterone sulfate decreased significantly. Serum follicle stimulating hormone (FSH) and prolactin levels did not change. LH-to-FSH ratio markedly decreased. SHBG level increased to the control level. After discontinuation of the pill, menstrual disturbances recurred in all patients. It appears that low-dose combined OCs containing DG is suitable for use in women with PCOD. However, long-term treatment may be needed since the abnormal menstrual pattern returned after stopping the pill. PMID- 2965637 TI - [Discovery of a complex t-haplotype in the gene pool of the synanthropic population of Mus musculus in Kishinev]. PMID- 2965638 TI - [Percutaneously implantable balloon-inflatable vascular prostheses. Initial clinical results]. AB - Restenosis and occlusion of atherosclerotic arteries are prevented by a newly developed arterial prosthesis consisting of a pliable and elastic wire mesh tube positioned to the balloon of an angioplasty catheter. As the balloon is inflated the prosthesis dilates and provides the arterial wall with a new mechanical support. This new device was successfully implanted in ten patients having an iliac artery stenosis and improved the therapeutic result of the preceding angioplasty. There were no complications. PMID- 2965639 TI - [Myelography after computed myelography for exploration of lumbar pain]. PMID- 2965640 TI - Purification and characterization of a novel protein from bovine aorta that inhibits coagulation. Inhibition of the phospholipid-dependent factor-Xa catalyzed prothrombin activation, through a high-affinity binding of the anticoagulant to the phospholipids. AB - A novel inhibitor of blood coagulation has been isolated from the intima of bovine aorta. The inhibitor, vascular anticoagulant (VAC), has been purified to an active fraction that contains two Coomassie-blue-staining bands (Mr = 34,000 and Mr = 32,000, as judged by sodium dodecyl sulfate/polyacrylamide electrophoresis). Both bands are single-chain proteins, having no glycoprotein features. Furthermore, they do not contain any detectable 4-carboxyglutamic acid residues. Both proteins have an identical isoelectric pH of approximately 4.5. VAC binds in the presence of calcium ions to a bilayer consisting of 20% dioleoylglycerophosphoserine and 80% dioleoylglycerophosphocholine with a Kd = 6 nM. The binding is dependent on the calcium concentration: half-saturation of binding occurs at a calcium concentration of 0.8 mM. The binding is completely reversible with EDTA. Furthermore the phospholipid/VAC ratio at saturation was n = 112 and n = 32 mol/mol for 0.5 mM Ca2+ and 2 mM Ca2+, respectively. Binding does not occur between VAC and pure dioleoylglycerophosphocholine. In a system with purified coagulation factors VAC inhibits the activation of prothrombin by factor Xa and calcium only in the presence of negatively charged phospholipids. VAC decreases the Vmax and increases the Km of the factor-Xa-catalyzed prothrombin activation. Based on these results, we conclude that we have purified from bovine aortic intima an anticoagulant protein, which exerts its activity through a calcium-dependent binding to negatively charged phospholipids, and thus interferes with the assembly of prothrombinase on the phospholipid surface. PMID- 2965641 TI - Does rat liver 6-phosphofructo-1-kinase exhibit 'half-of-the-sites phosphorylation'? AB - 6-Phosphofructo-1-kinase (PFK-1) from a variety of species and organs can undergo phosphorylation by cAMP-dependent protein kinase. In most studies the stoichiometry of the phosphorylation reaction was far below the expected minimum value of 4 mol phosphate/mol PFK-1 tetramer. The present study with rat liver PFK 1 and purified catalytic subunit of cAMP-dependent protein kinase was undertaken in order to find the maximum phosphorylation stoichiometry under well-defined conditions. Irrespective of whether PFK-1 had been first treated with purified protein phosphatase 2C or not, no more than 1.66 +/- 0.22 mol phosphate/mol PFK-1 tetramer was incorporated, the highest single value being 2 mol phosphate/PFK-1 tetramer. This stoichiometry was found to be independent from the method of protein evaluation (gel dye-binding assay or amino acid analysis) and from the concentration of PFK-1 in the phosphorylation system (15.6 nM-0.53 microM). The stoichiometry was not affected by the presence of allosteric ligands, fructose 1,6-bisphosphatase or the PFK-1-inactivating protein. The possibility could be excluded that partial proteolysis was responsible for the incomplete phosphorylation. Two-dimensional polyacrylamide gel electrophoresis gave no indication of the existence of two different subunits in rat liver PFK-1. Possible reasons why rat liver PFK-1 undergoes 'half-of-the-sites' phosphorylation are discussed. PMID- 2965642 TI - Reactivity of Ly-2+ T cells against 2,4,6-trinitrophenyl (TNP)-modified syngeneic stimulator cells: specificity, frequency of interleukin 2-producing Ly-2+ helper T cells and clonal segregation from Ly-2+ cytotoxic T lymphocytes. AB - The in vitro reactivity of purified murine Ly-2+ and L3T4+ T cells towards 2,4,6 trinitrophenyl (TNP)-modified syngeneic stimulator cells was analyzed. Both T cell subpopulations autonomously proliferated and produced interleukin 2. In either the Ly-2+ or L3T4+ T cell subset the frequencies of TNP-specific interleukin 2 (IL 2)-producing T lymphocyte precursors (IL 2 TL-p) were equally high (f = 1/400-1/1000). Clonally developing IL 2 TL of either T cell subset showed an exquisite antigen (TNP) specificity as shown by the split culture approach. TNP-specific Ly-2+ IL-2 TL used class I MHC (H-2Kk) gene products as major histocompatibility complex (MHC) restriction elements, while L3T4+ IL 2 TL proved to be class II MHC (H-2I-AkI-Ek) restricted. Clonal segregation analyses revealed that the majority of clonally developing TNP-reactive Ly-2+ TL segregated into either IL 2 TL-p or cytotoxic T lymphocyte presursors, i.e. both functions appear to be mutually exclusive. Less than 10% of the responding Ly-2+ T cells seemed to be bifunctional. These findings provide compelling evidence for the L3T4+ T cell-independent, autonomous reactivity of Ly-2+ T cells in MHC restricted antigen-specific responses and suggest T-T cell interactions within the functional heterogenous Ly-2+ T cell population. PMID- 2965643 TI - Soluble CD23 is released by B lymphocytes cycling in response to interleukin 4 and anti-Bp50 (CDw40). AB - An enzyme-linked immunoassay was developed to quantitate the production of soluble CD23 from cycling B lymphocytes. This molecule has identity both with B cell-derived B cell growth factor and with an IgE-binding factor. B lymphocytes, which had been stimulated for 3 days with phorbol dibutyrate and calcium ionophore, washed and recultured, failed to produce detectable levels of CD23 over the following 3 days. Soluble CD23 was found, however, in the supernatant of cultures where recombinant interleukin 4 had been included. The level of CD23 rose dramatically when anti-Bp50 had also been added. By contrast, anti-Bp50, alone or together with low molecular weight T cell-derived B cell growth factor, failed to promote the release of CD23 in detectable amounts. There was a strong correlation between the appearance of soluble CD23 in culture supernatants and the expression of CD23 on the surface of restimulated cells. The level of CD23 release appeared to relate more to the continued cycling of cells than to their differentiation to immunoglobulin secretion. These findings are discussed with particular emphasis on the role of CD23 as an important multi-functional lymphokine in B lymphocyte physiology. PMID- 2965644 TI - T helper cell-dependent induction of resting B cell differentiation need not require cognate cell interactions. AB - We have analyzed the role of cognate interaction with helper T cells (Th) in support of resting B cell differentiation to plaque formation. Co-culture of histoincompatible resting B cells and resting Th cells resulted in the induction of plaque-forming cells when dimeric but not monomeric fragments of anti-T cell receptor (TcR) antibody were added to culture. The efficiency of B cell activation was comparable to that supported by lipopolysaccharide and lectin mediated Th-B cell conjugate formation. Further, if resting Th cells were preactivated with antigen and histocompatible antigen-presenting cells, the requirement for addition of anti-TcR to mixtures of histoincompatible Th and B cells was obviated. These results demonstrate that TcR-mediated Th recognition of major histoincompatibility complex class II/antigen composites on the resting B cell membrane does not provide obligate signals for B cell differentiation to plaque formation. We are left with two possibilities. Either the entire process of Th cell-dependent induction of resting B cell differentiation is mediated by soluble lymphokines or if Th-B cell contact is mandatory, it is mediated through nonpolymorphic cell surface determinants. PMID- 2965645 TI - The molecular interactions with helper T cells which limit antigen-specific B cell differentiation. AB - Helper T (Th) cell-dependent activation requirements for 2,4,6-trinitrophenyl (TNP)-specific resting B cells obtained from mice transgenic for Sp-6 mu, kappa genes were analyzed. Carrier-specific T cell help required linked recognition of TNP carrier and was functionally restricted by the B cell major histocompatibility complex. However, histoincompatible T cell-B cell conjugates formed by bridging surface immunoglobulin and Th cell receptor for antigen (TcR) through TNP-conjugated anti-TcR antibodies resulted in the efficient differentiation of TNP-specific B cells. Thus, Th cell-dependent cognate recognition of B cells is not obligatory. Specific conjugate formation could be obviated by using unconjugated fragments of anti-TcR antibodies. If dimeric, these fragments supported the Th cell-dependent differentiation of co-cultured histoincompatible resting B cells. Unconjugated monomeric fragments were ineffective, demonstrating the necessity for TcR cross-linking. Resting B cells from Sp-6+ mice rendered TNP-conjugated monomeric fragments of anti-TcR antibodies effectively multivalent, thereby satisfying conditions for the activation of co-cultured Th cells. The results demonstrate that Th cells do not transduce activation signals through TcR recognition of B cell membrane associated ligand which limit the induction of B cell differentiation. Cross linking of TcR on Th cells is required, sufficient and can be induced through interaction with the antigen-specific B cell surface. PMID- 2965646 TI - A noncognate interaction with anti-receptor antibody-activated helper T cells induces small resting murine B cells to proliferate and to secrete antibody. AB - Culture of small resting allogeneic B cells (of an irrelevant haplotype) with two clones of T helper (Th) cells that were activated by the F23.1 anti-T cell receptor antibody led to the activation of B cells to proliferate and to secrete antibody. Th cell supernatants by themselves had no effect on resting B cells (even in the presence of intact F23.1 antibody), but could induce antibody secretion by anti-Ig-preactivated B cells. Both F23.1+ clones (E9.D4 and 4.35F2) and one F23.1- clone (D2.2) could synergize with supernatants from activated E9.D4 T cells to induce B cell activation. F(ab')2 fragments of F23.1 induced E9.D4 to activate B cells as efficiently as intact F23.1 and B cell populations that had been incubated with F23.1 were not activated when cultured with E9.D4, although T cells recognized cell-presented F23.1 and were weakly activated. Reduction of the density of F23.1 adsorbed to plastic resulted in weak T cell activation, and these T cells did not induce B cell responses. Haptenated B cell populations, although recognized by E9.D4, were not activated. Separation of T and B cells by a 0.4-micron membrane prevented T-dependent B cell activation, although Th cell-derived B cell-activating lymphokines would be assayed across these membranes. These results suggest a polyclonal noncognate B cell activation that depends on physical contact between B cells and activated T cells. The requirement for a cognate interaction of Th with B cells for the production and delivery of B help can therefore be overcome by activating Th cells with high densities of T cell receptor ligands. PMID- 2965647 TI - A significant proportion of normal resting B cells are induced to secrete immunoglobulin through contact with anti-receptor antibody-activated helper T cells in clonal cultures. AB - This report describes single-cell techniques to address the nature of a cellular interaction in which activated T lymphocytes stimulate small resting B cells to develop into antibody-forming cell clones in the absence of any surface immunoglobulin ligand or an antigen bridge. The cloned T helper cell line E9.D4 was stimulated with the anti-V beta 8 antibody F23.1 bound to the plastic of Terasaki 10-ul culture wells. When an excess of T helper lymphocytes was used (1,000 X-irradiated or 600 unirradiated, stimulated E9.D4 cells), 10-25% of B cells responded by antibody formation as judged by an enzyme-linked immunosorbent assay performed after 5 days of culture. When one of a very small number of B cells were present, the rate-limiting step to antibody-forming cell formation was the number of T cells present. Far fewer T cells sufficed for stimulation when culture trays were tilted to force T and B cells into proximity at the sulcus formed at the bottom edge of the culture wells. When T cell numbers were limiting, unirradiated T cells out-performed irradiated T cells. Some cell clones held for 7 days switched to IgG antibody production. E9.D4 supernatants were virtually ineffective in causing B cell stimulation, even when 3T3 filler cells were added to support cultures. The results suggest that cell contact, and perhaps conjugate formation, with a strongly activated T cell can cause changes in the adjacent resting B cells akin to those of Ig receptor cross-linking, following which a lymphokine flux (even one not involving IL 4 and 5) promotes antibody-forming cell development. PMID- 2965648 TI - Analysis of CD3 and antigen receptor expression on T cell subpopulations of aged athymic mice. AB - The expression of the T cell antigen receptor on subpopulations of extrathymically generated T cells from athymic mice was investigated and compared to antigen receptor expression in normal mice. To this end, spleen and lymph node cells from 18 individual athymic BALB/c nu/nu mice between 6 and 12 months of age and from normal controls were enriched for T cells by nylon wool filtration. Expression of the following cell surface markers was analyzed by two-color flow cytometry: Thy-1, CD4, CD8, V beta 8 and CD3. The distribution of subpopulations as defined by these markers varied much more among athymic than among normal mice. Some recurrent patterns did, however, occur that may be characteristic of the extrathymic pathway of T cell differentiation in nu/nu mice. Among Thy-1.2+ cells, CD8+ cells predominated over CD4+ cells. No CD4+8+ "double positive" cells were found, but CD4-8- "double negative" cell constituted 16% on average. All nu/nu nylon wool-nonadherent cells expressing Thy-1 at a normal level also expressed CD3, whereas Thy-1low and Thy-1- cells were CD3-. The fraction of V beta 8 T cells among the CD4+ and CD8+ subsets was near to normal in the majority of these animals. Most interestingly, the density of V beta 8 and CD3 expression was lower in athymic than in euthymic animals. This level of T cell antigen receptor expression was, however, higher than on immature CD4+8+ thymocytes. A fraction of the nude T cells presently characterized responded with proliferation to both anti-T cell receptor V beta 8 monoclonal antibody and to concanavalin A. Despite their apparently normal phenotype (with the exception of reduced T cell receptor expression), this response was, however, 4 to 10 times smaller than that of normal control cells. The presently described Thy-1+ T cell receptor intermediate cells may either be a(n) aberrant lineage(s) only generated extrathymically, or represent the accumulation of an immature intermediate stage of normal (i.e. thymic) T cell differentiation. PMID- 2965649 TI - Growth-dependent variation of major histocompatibility complex (MHC) restriction and expression of Ly-2 and CD3/alpha/beta T cell receptor in cloned cytotoxic T cells. AB - The specificity of major histocompatibility complex (MHC)-restricted antigen recognition by cytotoxic T cells (CTL) has been clearly correlated to the alpha/beta T cell receptor (TcR) complex on the T cell surface. Occasional changes in the specificity of in vitro cultivated CTL clones, therefore, have been suspected to result from alterations of the genes coding for the TcR alpha and/or beta chain. Here we demonstrate that pronounced variations in the stringency of MHC restriction, previously reported to occur during long-term culture of 2,4,6-trinitrophenyl (TNP)-specific CTL clones, may occur rapidly in a growth-dependent, reversible manner, i.e. without structural TcR variation. Several H-2b TNP-specific CTL clones were shown to possess strong cross reactivity for H-2k TNP target cells when seeded at low cell numbers, but exhibit reduced or undetectable cross-reaction to H-2k TNP in high-density cultures. Another clone revealed "heteroclitic" properties with significantly stronger cytotoxic activity towards allogeneic (H-2k) than syngeneic (H-2b) TNP-modified target cells. In this case dilute cultures appeared as exclusively allo-MHC restricted, whereas dense cultures were allo/self cross-restricted. In all instances these phenomena were accompanied by cell density-dependent quantitative changes in the expression of Ly-2 and T cell antigen receptor. CTL from dilute cultures had at least 2-fold higher surface concentrations of Ly-2 and CD3 antigens than cells from dense cultures while other surface markers such as Thy-1 or LFA-1 were completely identical. No such effects were observed for CTL clones exhibiting cell density-independent specificity patterns. We conclude from these findings that (a) the stringency of MHC restriction specificity may be significantly affected by the amount of expressed TcR and/or Ly-2 molecules, (b) CTL possess mechanisms to regulate Ly-2 and TcR expression and, hence, their MHC restricted antigen recognition, and (c) the ability to regulate Ly-2 and TcR expression may be altered during prolonged culture of a CTL clone. PMID- 2965650 TI - Induction of human in vitro IgE synthesis via stimulation by anti-CD3 antibody. AB - Mitogens such as pokeweed mitogen, Staphylococcus aureus Cowan I, concanavalin A and phytohemagglutinin and monoclonal anti-CD antibodies were examined for their capacity to induce IgE synthesis by peripheral blood leukocytes obtained from both atopic and nonatopic individuals. While lectins failed to induce IgE synthesis, monoclonal anti-CD3 antibodies were very potent stimuli for the induction of human in vitro IgE synthesis, possibly due to the activation of T cells. Activation via CD4 or CD8 molecules by OKT4 or OKT8 antibodies did not lead to a T cell-dependent modulation of IgE synthesis. PWM acted in synergy with the enhancing effect of anti-CD3 antibodies for IgE synthesis. In vitro IgG production was less affected. These results indicate that activation of T cell via CD3 molecules may be important in the regulation of IgE immune responses in man. Furthermore, the successful induction of IgE synthesis by anti-CD3 antibody in unfractionated peripheral blood leukocytes culture provides a simple model for investigation of human IgE regulatory mechanism in vitro. PMID- 2965651 TI - The major surface protein of Leishmania promastigotes is a fibronectin-like molecule. AB - The major surface glycoprotein of Leishmania chagasi promastigotes showed crossreactivity with fibronectin (Fn), a large glycoprotein that is a major constituent of the extracellular matrix of most mononuclear cells. Polyclonal and monoclonal antibodies against Fn precipitated two molecules of 63-58 kDa from the lysates of both 125I and [35S]methionine-labeled promastigotes. In addition, a monoclonal antibody against a 15-kDa fragment of Fn containing the Arg-Gly-Asp Ser (RGDS) sequence and several polyclonal monospecific mouse antibodies against a synthetic RGDS peptide also recognized the above two molecules. The attachment of Leishmania promastigotes to mouse peritoneal macrophages in vitro was partially inhibited when promastigotes were treated with F(ab')2 fragment of an anti-Fn IgG. Identical results were obtained by saturating the Fn receptors on macrophages using different peptides containing the RGDS sequence. Moreover, antigen preparations rich in glycoprotein 63 could efficiently promote the attachment and spreading of 3T3 mouse fibroblasts to surfaces coated with the antigen. These results clearly suggest that the gp63 of L. chagasi promastigotes is an Fn-like molecule that shares certain biological and molecular characteristics with Fn. PMID- 2965652 TI - Anti-suppressor effect of cyclophosphamide on the development of spontaneous diabetes in NOD mice. AB - In the NOD mouse, an autoimmune process beginning by 5 weeks of age with lymphocyte infiltration and destruction of insulin-secreting beta cells leads to overt diabetes which begins to appear by 11 weeks of age. Although there is a high incidence of insulitis by 10 weeks of age (greater than 80%) in both males and females, by 30 weeks of age diabetic symptoms have occurred in 53-80% of females and in 12-40% of males. Intraperitoneal injection of a high dose (200 mg/kg) of cyclophosphamide (CY) consistently induces the onset of diabetes in male and female NOD mice at an age when spontaneous diabetes rarely occurs. Spleen T cells from CY-induced diabetic mice are capable of transferring the disease into irradiated nondiabetic syngeneic recipients. This indicates that the diabetogenic effect of CY is not mediated by direct toxicity on pancreatic beta cells but is mediated by abrogation of a suppressor mechanism which may prevent activation of T cells responsible for the development of diabetes in the NOD mouse. Additionally, CY is only effective in NOD mice and not in F1 hybrids between NOD and other strains of mice. Thus, the potential beta cell aggressor mechanism is not present in these hybrids as it is in homozygous mice, which indicates that it is not under the control of dominant genes. PMID- 2965653 TI - The role of protein kinase C in early activation vs. growth of T lymphocytes. AB - Protein kinase C (PKC) has been implicated in the signaling of a number of cellular responses including activation of T cells. In an approach to study the role of PKC in both early activation and interleukin 2 (IL2)-dependent growth, we have used in vivo phosphorylation of endogenous substrates as a marker for PKC activation. In the present report the requirements for CD3-specific activation of PKC are defined, and it is shown that the conditions required for optimal activation coincide with the induction of a proliferative response. Moreover, it is also shown that IL2 receptor interactions induce only low levels of PKC substrate phosphorylation and that IL2 does not cooperate with CD3-specific signals on the level of PKC activation. PMID- 2965654 TI - Effects of triflusal and acetylsalicylic acid on platelet aggregation in whole blood of diabetic patients. AB - A study was made on the inhibitory effect of triflusal (600 mg/d X 15) and acetylsalicylic acid (ASA, 400 mg/d X 15) on platelet aggregation in whole blood (WB) and platelet-rich plasma (PRP) induced by ADP (2.5 mumol/l), adrenaline (50 mumol/l), collagen (1 microgram/ml) and arachidonic acid (0.8 mmol/l), in 30 insulin-dependent diabetic patients without vascular complications. Determination was also made of the serum levels of thromboxane B2 (TxB2) and of the plasma levels of 6-keto-PGF1-alpha and of beta-thromboglobulin (B-TG). Both drugs exhibited higher inhibitory effects in WB than in PRP. In WB, a significant difference between triflusal and ASA was observed against ADP-induced aggregation (67% and 46% inhibition respectively, p less than 0.01). Both drugs strongly inhibit the formation of TxB2 in serum (85% and 99%, respectively). Triflusal does not significantly change the plasma levels of 6-keto-PGF1-alpha; ASA, by contrast, causes reduction of over 95% in those plasma levels. The plasma levels of B-TG were not modified by either of the drugs. PMID- 2965655 TI - [Organization of the work of a medical prevention service for animal husbandry workers]. PMID- 2965656 TI - [Effect of alpha-tocopherol and thiamine on the heart pumping function in hypertrophy]. PMID- 2965657 TI - [Role of brain serotonin in the regulation of sexual maturation in male rats]. AB - The effect of decrease of the brain serotonin level in different periods of prepubertal development on sexual maturation of male rats, was studied. Injections of p-CPA on the 30th and 34th days of life activated the endocrine function of testis in 60-day old animals. Injections of p-CPA later in life, on the 40th and 44th days, delayed sexual maturation as revealed by a decrease of spermatogenesis and endocrine function of testis. The data obtained suggest that serotonin has a dual role in regulation of the rate of sexual maturation in different periods of prepubertal age. PMID- 2965658 TI - [Pellagra-like changes following glibenclamide treatment in a patient with diabetes mellitus and vitiligo]. PMID- 2965659 TI - Fructose 2,6-bisphosphate-dependent regulation of phosphofructokinase in rat submandibular gland. AB - 1. Regulation of phosphofructokinase in rat submandibular gland was non-Michaelis Menten type at physiological pH. 2. At pH 7.3, ATP played a dual role on phosphofructokinase acting as a substrate and inhibitor at high concentration of ATP. 3. The activator of phosphofructokinase was present in cytosol fraction, and its properties were resemble to those of fructose 2,6-bisphosphate. 4. Both the activator and authentic fructose 2,6-bisphosphate relieved the inhibition of phosphofructokinase by ATP, and increased the affinity for fructose 6-phosphate. 5. Concentration of fructose 2,6-bisphosphate in rat submandibular gland was 8.22 nmol/g tissue, and which was about the half of that in liver. 6. Phosphofructokinase in rat submandibular gland was found to be regulated synergistically by ATP, fructose 6-phosphate and fructose 2,6-bisphosphate. PMID- 2965660 TI - Full oestrogenic activity of C19-delta 5 adrenal steroids in rat pituitary lactotrophs and somatotrophs. AB - Oestrogens are known to exert specific stimulatory effects on basal and dopamine (DA)-inhibited prolactin (PRL) release as well as on PRL cell content in rat adenohypophysial cells in primary culture. Recently, we have demonstrated that classical oestrogens increase growth hormone (GH) release and cellular GH content in rat pituitary somatotrophs. Since there is evidence that 5-androstene-3 beta, 17 beta-diol (delta 5-diol), a metabolite of dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEA-S) can induce typical oestrogenic responses in target tissues, we have investigated the effect of C19 adrenal steroids and compared them to that of 17 beta-oestradiol (E2) on the above-indicated parameters. Following a 72-h incubation, maximal concentrations of E2, delta 5-diol and DHEA caused a similar increase in PRL cell content at respective EC50 values of 0.020, 14 and 115 nM. The sensitivity of lactotrophs to DA action was decreased by approximately 4-fold after a 48-h exposure to maximal concentrations of delta 5-diol or DHEA. The time course of the antidopaminergic effect of delta 5-diol and DHEA was almost superimposable to that of E2. A 72-h incubation with 5 microM DHEA-S, a concentration within the range of blood levels found in women, doubled (P less than or equal to 0.001) cellular PRL accumulation. On the other hand, androstenedione (delta 4-dione) was without effect on any of the parameters measured. All stimulatory effects induced by maximal effective concentrations of delta 5-diol, DHEA or DHEA-S were competitively inhibited by simultaneous incubation with the antioestrogen LY156758. The amplitude of the stimulatory effects of delta 5-diol and DHEA on GH cell content as well as on spontaneous GH releasing factor (GRF)-induced GH release was superimposable to that observed with E2. The effect of the steroids on GH cell content was exerted at 0.016 nM (E2), 12 nM (delta 5-diol) and 250 nM (DHEA). Simultaneous incubation with LY156758 completely blocked the effect of maximally effective concentrations of E2, delta 5-diol and DHEA in somatotrophs. Furthermore, a physiological concentration of DHEA-S (5 microM) enhanced spontaneous as well as GRF-induced GH release. On the other hand, delta 4-dione as well as testosterone and dihydrotestosterone did not alter GH release. The present data demonstrate that delta 5-diol, DHEA and DHEA-S can exert full oestrogenic effects in lactotrophs and somatotrophs, thus supporting their potential oestrogenic role under both physiological and pathological conditions. PMID- 2965661 TI - [Diabetic cardiomyopathy: preclinical pathology disclosed by the increase in ventricular afterload]. AB - In order to test the importance of the association of diabetes mellitus and arterial hypertension in generating morphological and functional changes of the left ventricle (LV) consistent with a cardiomyopathy, 37 patients, aged 27 +/- 6.7 years, were studied by standard and digitized M-Mode echo: eighteen of them were affected by diabetes mellitus, 11 by arterial hypertension, 8 by diabetes and hypertension. Each group was compared to the others and with a group of 14 normal subjects. In order to verify the importance of increased ventricular after load in modifying ventricular performance of diabetic patients, changes of the peak rate of systolic and diastolic variation of LV diameter and changes of the peak rate of interventricular septum and posterior wall excursion (IVSE, PWE) were evaluated after methoxamine hydrochloride infusion in 8 diabetic and 6 normal subjects. In diabetics the ratio between ventricular thickness and diameter (h/r) was greater than normal subjects (p less than 0.02); this ratio resulted higher in patients with diabetic rhinopathy who also exhibited an isovolumic diastolic period longer than normal (p less than 0.02). Both h/r ratio and isovolumic diastolic period (IDP) were higher in diabetic-hypertensive group as compared to normals (p less than 0.001), strictly diabetic (p less than 0.01 and p less than 0.001) or hypertensive subjects (p less than 0.01). Diabetic hypertensive group, exhibited a lowering of the systolic and diastolic peak rate of IVSE (p less than 0.01) as well as of systolic peak rate of PWE as compared to the other three groups (p less than 0.05). PMID- 2965662 TI - [Medical genetic study of the population of Kostroma Province. V. The hereditary pathology burden of urban and rural populations]. AB - Medical genetic study was carried out in the urban and rural populations of Kostroma Province. Urban populations were shown to have lower frequencies of "rare" forms of autosomal recessive diseases, in comparison with those in the rural populations. Analysis of interrelationship between genetical structure of populations and prevalencies of hereditary diseases in the populations revealed clear relations between the load of autosomal recessive diseases and the level of inbreeding in the populations. PMID- 2965663 TI - The araC gene of Citrobacter freundii. AB - The araC gene of Citrobacter freundii was cloned into plasmid pBR322 and expressed in Escherichia coli and Salmonella typhimurium. The nucleotide sequence and the predicted translational product were determined and compared to those of E. coli, S. typhimurium and Erwinia carotovora. The predicted translational product is 281 amino acids (aa) long, identical in size to that of S. typhimurium, and is 11 and 29 aa shorter than that of E. coli and E. carotovora, respectively. The nucleotide sequence of the araC gene of C. freundii is 83% homologous to the araC genes of both E. coli and S. typhimurium, but only 60% homologous to that of E. carotovora with respect to the regions they share. The predicted amino acid sequence is highly conserved and shows 96% and 94% homology to S. typhimurium and E. coli, respectively. E. carotovora shows only a 58% aa homology. The activator and autoregulatory activities of each plasmid encoded AraC protein in a S. typhimurium araC::lacZ protein fusion strain were examined. PMID- 2965664 TI - Nucleotide sequence and characterization of a gene encoding the phytochrome polypeptide from Avena. AB - We have isolated and characterized a gene encoding the phytochrome polypeptide of Avena. Based on nucleotide sequence identity with previously sequenced cDNA clones this gene is designated as type 3 (phy3). The gene is about 5.9 kb long with six exons and five introns, one each of the latter in the 5' and 3' untranslated regions. The largest exon encodes the entire 74-kDa, chromophore bearing, N-terminal domain of the photo-receptor postulated to be directly involved in its mechanism of action. The transcription start point, identified by mung-bean nuclease digestion, is located 24 to 35 bp downstream from a tandem TATA box. Sequence elements homologous to a number of motifs implicated as upstream regulatory elements in other genes are present in the 5'-flanking DNA of phy3. Particularly intriguing are three elements at positions -140, -470 and 650. These elements share homology with the 'GT' motif postulated to be a component of the light-regulatory element of genes encoding the small subunit of ribulose bisphosphate carboxylase. PMID- 2965665 TI - Evidence for prokaryotic transcription and translation control regions in the human factor IX gene. AB - A human factor IX cDNA clone isolated from a liver cDNA library constructed in phage lambda gt11 vector was shown to express factor IX protein in Escherichia coli. A factor IX immunospecific protein of 46.8 kDa was expressed, but was not a beta-galactosidase-factor IX fusion protein. Expression was seen when the factor IX cDNA was cloned into two different vector systems, lambda gt11 and pUC9, in both orientations with respect to the vector lacZ promoter. The expression of factor IX was not under control of the lacZ promoter of either vector system. In addition, when the factor IX cDNA fragment was subcloned in both orientations into a promoterless cloning vector (p CPP3), the factor IX cDNA fragment demonstrated promoter activity when inserted in only one orientation resulting in expression of chloramphenicol acetyl transferase in E. coli and Bacillus subtilis. A DNA computer search of the N-terminal sequences of the factor IX gene revealed prokaryotic-like promoter and ribosome-binding site (RBS) sequences with strong homology to the E. coli consensus sequences. The predicted sites homologous with prokaryotic promoter and RBS consensus sequences are followed by an in-frame methionine which could correspond to the translation start codon of the expressed factor IX. This report provides the first evidence that a eukaryotic gene encodes the information necessary for both transcription and translation to control gene expression in a prokaryotic host. PMID- 2965666 TI - [Sanitary and hygienic working conditions and characteristics of chronic dust induced bronchitis in miners in the coal shafts of the Podmoskva basin]. PMID- 2965667 TI - [Experience of the Department of Industrial Hygiene with cooperative work in practical public health]. PMID- 2965668 TI - Don Peterson: a man, a computer, and miracles. PMID- 2965669 TI - [Primary health care]. PMID- 2965670 TI - Accountability in health nursing. PMID- 2965671 TI - Accountability in nursing practice. PMID- 2965672 TI - [Targets for health for all--WHO]. PMID- 2965673 TI - Effective approach to conflict resolution in management. PMID- 2965674 TI - Hospital acquired infection--an introduction. PMID- 2965675 TI - [Made to measure]. PMID- 2965676 TI - Diabetes-induced cholestasis in the rat: possible role of hyperglycemia and hypoinsulinemia. AB - In one of our earlier studies, an impaired biliary function in diabetes was suggested. We studied bile formation in rats with streptozotocin-induced diabetes (60 mg per kg body weight). Diabetic rats showed hyperglycemia and hypoinsulinemia, but no significant changes in hematocrit, plasma protein concentration or plasma osmolality. Bile flow was significantly (p less than 0.05) reduced (-23%) as compared with control animals, despite a higher (p less than 0.05) bile acid secretion rate (+56%). The biliary responses to three choleretic compounds (taurocholate, ursodeoxycholate and insulin), acting in a very different way upon bile formation, were not impaired in diabetes. The study of the relationship between bile acid output and bile flow, after infusion of taurocholate at different doses (0.25 to 1.5 mumoles per min per 100 gm body weight) showed that diabetes-induced cholestasis in the rat is mainly related to a decreased bile acid-independent fraction of the bile flow. We tested the possible role of hyperglycemia and hypoinsulinemia as cholestatic factors in diabetes. Glucose infusion [300 mM, 150 microliter per min (Group G)] induced a significant (p less than 0.05) reduction in bile flow (-0.33 microliter per min per gm liver) as compared to the basal period. After acute pancreatectomy (P) or mannoheptulose treatment [0.14 mmole per 100 gm body weight (Group M)], similar cholestatic effects were observed (-0.29 and -0.27 microliter per min per gm liver, respectively). However, plasma glucose and insulin concentrations were higher (p less than 0.01) in Group G than in P or M.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965677 TI - Pleural effusion due to dracunculosis. PMID- 2965678 TI - Effects of moderate diabetes on cardiac performance in spontaneously hypertensive and Wistar-Kyoto rats. AB - To assess the effects of imposition of moderate diabetes on in vivo cardiac performance in gradually proceeding hypertension, spontaneously hypertensive (SHR) and Wistar-Kyoto rats (WKY) were treated with streptozotocin (40 mg/kg) or vehicle at 8 weeks of age. Four and 20 weeks later, with the rats under ether anesthesia, peak cardiac output and stroke volume were measured during volume loading and peak left ventricular developed pressure and maximum rate of rise of pressure (dP/dtmax) were determined during aortic occlusion. Additionally, passive pressure-volume relations were obtained during saline infusion in potassium-arrested hearts, and the chamber stiffness constant was derived from one exponential function. There was a mortality of 16.1% in the diabetic SHR only. While basal and stressed cardiac performance was unchanged despite the already decreased mean arterial pressure and left ventricular weight at 4 weeks, the diabetic SHR revealed significant decreases in peak cardiac pumping indexes, peak left ventricular developed pressure, and dP/dtmax, with unchanged resting cardiac function, at 20 weeks. Changes seen in the diabetic WKY were reduced left ventricular weight at 4 weeks and reduced peak left ventricular dP/dtmax at 20 weeks. The chamber stiffness was unaltered with strain or diabetes. These data show that imposition of even moderate diabetes substantially influences the stress-loaded in vivo cardiac performance in the SHR, whereas it produces only minor changes in the WKY. PMID- 2965679 TI - Employer/employee rights and privileges. PMID- 2965680 TI - Inactivation of suppressor T-cell activity by nontoxic monophosphoryl lipid A. AB - Treatment with nontoxic monophosphoryl lipid A (MPL), which was derived from a polysaccharide-deficient, heptoseless Re mutant of Salmonella typhimurium, was found to inactivate suppressor T-cell activity, as evidenced by a decrease in the degree of low-dose immunological paralysis expressed and an increase in the magnitude of the antibody response to type III pneumococcal polysaccharide. The effects produced, which could not be attributed to the polyclonal activation of immune B cells by MPL, were dependent upon the dose of MPL used, as well as the time when MPL was given relative to low-dose priming or immunization with type III pneumococcal polysaccharide. Neither amplifier nor helper T-cell activity was decreased by treatment with the same, or larger, doses of MPL. The significance of these findings to the use of MPL as an immunological adjuvant or an immunomodulating agent is discussed. PMID- 2965681 TI - Characterization of an in vitro-stimulated, Cryptococcus neoformans-specific second-order suppressor T cell and its precursor. AB - Using a cryptococcal culture filtrate antigen (CneF) in a murine model, we have demonstrated previously that a cascade of Cryptococcus neoformans-specific suppressor T cells and soluble factors function in suppressing the cryptococcal delayed-type hypersensitivity (DTH) response. In addition, we have successfully hybridized the C. neoformans-specific, first-order T-suppressor (Ts1) cell and have established that the culture supernatant (hTsF1) from this hybridoma induces second-order T-suppressor (Ts2) cells in vivo. Here we report the in vitro induction of expression-phase suppressor cells. The suppressor cells were induced by culturing nylon wool-nonadherent splenic cells from naive mice with hTsF1 in the absence of CneF. Nylon wool-nonadherent splenic cells similarly cultured with supernatants from the BW5147 thymoma cells, the fusion partners of the hybridoma, did not significantly suppress the cryptococcal DTH response. The suppressor cells were designated Ts2 cells based on their similarities in function, specificity, and phenotype, i.e. L3T4-, Lyt-2+, and I-J+, to the in vivo-induced Ts2 cells. By employing the in vitro culture technique, we demonstrated that the precursors of the functional Ts2 cells were L3T4- Lyt-1-2+ I-J- cells. The induction of Ts2 cells was not associated with [3H]thymidine incorporation; therefore, we concluded that hTsF1 induces the Lyt-2+ I-J- cells to differentiate into Lyt-2+ I-J+ functional Ts2 cells without a significant amount of proliferation. From the results of this study, a better understanding of the processes involved in the regulation of the DTH response to CneF was achieved. The in vitro culture technique will allow for further detailed studies of the interactions between the various cell populations and the Ts1 cell-derived soluble factor during the induction of Ts2 cells. PMID- 2965683 TI - IgE-isotype-specific suppressor cells in the mouse: characterization using tetraparental chimera mice of high (DBA/2) and low (SJL) IgE responder embryos. AB - Tetraparental chimera mice were developed by aggregation of IgE high responder (DBA/2) and IgE low responder (SJL) embryos. Anti-dinitrophenyl (DNP) IgE antibody response in such mice (SJL----DBA/2) upon challenge with DNP-keyhole limpet hemocyanin (KLH) in alum was clearly suppressed, while anti-DNP IgG antibody response was not. High-titer anti-DNP IgE and IgG antibody response developed in F1 hybrid mice of SJL and DBA/2 (SDF1) mice. The experimental results suggest that high IgE antibody production is the dominant trait, and the IgE-specific suppressor gene in SJL mice is autosomal recessive. IgE-specific suppressor T cells in SJL mice actively suppressed IgE antibody formation by DBA/2 immuno-competent cells across the histocompatibility barrier. Hapten specific B cells and carrier-specific T cells were prepared in SJL----DBA/2 and SDF1 mice by immunization with DNP-KLH or ovalbumin (OA) in alum and transferred to irradiated SDF1 mice followed by challenge with DNP-OA. Hapten-specific B cells and carrier-specific helper T cells clearly developed in SDF1 mice. Recipient mice transferred with DNP-KLH-primed SDF1 spleen cells and OA-primed SDF1 spleen cells showed high-titer anti-DNP IgE and IgG antibody responses. OA primed SJL----DBA/2 spleen cells cotransferred with DNP-KLH-primed SDF1 spleen cells and OA-primed SDF1 spleen cells completely abolished secondary anti-DNP IgE antibody response. The data suggest that carrier-specific helper T cells for IgE and IgG antibody responses are distinct. The regulatory role of IgE-isotype specific suppressor cells were considered to be the interference of cooperative cellular interaction between IgE B cells and carrier-specific, IgE-specific helper T cells. PMID- 2965682 TI - Effect of neonatal injection with antibodies to Leishmania mexicana on its growth in adult infected mice. AB - Mice inoculated with monoclonal antibodies (MAb) directed to Leishmania mexicana antigens were not protected from growth of a subsequent challenge infection; this was the case even when those antibodies were capable of inhibiting parasite growth in vitro. However F(ab')2 fragments of one antibody (1E1) were protective in vivo. When neonatal mice were injected with MAb and subsequently infected as adults, the animals were more susceptible to parasite growth than uninjected controls. This increased susceptibility could be adoptively transferred with Lyt 1+ cells. Separate groups of animals were immunized with different MAb to L. mexicana, and parasite growth in these animals was studied. In no case was parasite growth altered, though these mice did produce specific antibodies directed against the immunizing MAb (anti-idiotypic antibodies). When neonatal mice were injected with these latter reagents, they were found to be more resistant to challenge infection than control animals. This resistance was associated with an enhanced ability of spleen cells from these mice to produce, on stimulation with parasite antigens in vitro, a factor rendering normal macrophages cytocidal for L. mexicana. PMID- 2965684 TI - Cognitive functioning of juveniles with Down syndrome and other forms of mental retardation. AB - The performance of juvenile Down syndrome individuals and two groups of oligophrenes was compared in several cognitive tests. All three groups of subjects were of the same CA, the Down's and one group of mentally retarded non Down's had an average MA of 62, the other group an MA of 86 months. Though the group with the highest mental age usually outperformed the other two, significant differences were rare. It is concluded that at this age Down's and mentally retarded with different etiologies generally have rather similar cognitive abilities and that the intellectual deterioration of Down's, which at a later age inevitably leads to Alzheimer's disease, indeed starts only at a later CA. PMID- 2965685 TI - Action of the sympathetic system on skeletal muscle. AB - The action exerted by the sympathetic system on skeletal muscle is briefly reviewed with regard to the effects induced by the adrenergic mediator on extra fusal muscle fibre contractility and on muscle spindle receptor organs. In particular, the different responses of slow - and fast - contracting muscles to catecholamine administration and to sympathetic stimulation are reported. Also, the sympathetically--induced effects on muscle spindle afferent activity are discussed on the basis of recent anatomical and functional data suggesting the existence of a direct action exerted by noradrenergic axons on intrafusal muscle fibres. Some of the possible implications of these data are then briefly mentioned. PMID- 2965686 TI - Successful HLA-A, B, C, DR and DQ-Identical sibling kidney transplantation with mixed lymphocyte culture reactivity and positive B-cell cross-match. PMID- 2965687 TI - Splicing of messenger RNA precursors. PMID- 2965688 TI - Fibronectins: a family of complex and versatile adhesive glycoproteins derived from a single gene. PMID- 2965689 TI - [Infantile acropustulosis]. AB - Infantile acropustulosis, a dermatosis first described in 1979, is reviewed on the basis of the additional information provided by the 34 new cases that have appeared in the literature since then. This dermatosis of infancy consists of chronically relapsing, highly pruritic pustules, involving the extremities, chiefly the palms and soles. The etiology is unknown. Sulfones are the only effective form of therapy, but they have many side-effects and their use is only rarely indicated in this benign disease. The differential diagnosis, including many other pustular eruptions of infancy, is discussed in detail. PMID- 2965690 TI - [Photoallergy to Neotri and cross reaction to tenoretic--detection by systemic photoprovocation]. AB - A patient is presented who suffered for 3 years from increasing photosensitivity with chronic eczematous lesions in sun-exposed areas. He had taken one Neotri (triamterene, xipamide) tablet daily for 6 years. After discontinuation of the drug, phototesting and a photopatch test failed to reveal pathological reactions. Eczematous lesions, however, were induced in test areas upon systemic photochallenging with Neotri. One year later, after the antihypertensive medication had been changed from Adalat (nifedipine) to Teneretic (atenolol, chlortalidone) the eczematous photosensitive reaction recurred. Since both xipamide and chlortalidone have a chlorsulfamoyl-substituted aromatic ring in common, it seems that a photoallergic cross-reaction occurred. PMID- 2965691 TI - [Autoimmunity in alopecia areata. An assessment in 100 patients]. AB - The sera of 100 patients with varying degrees of alopecia areata were examined for the presence of circulating autoantibodies against nuclear, mitochondrial, reticulin-fibril, smooth muscle, parietal cell, thyroglobulin, and microsomal components. Furthermore, in all cases we measured the percentage of total T lymphocytes, T-helper, and T-suppressor cells. Other diseases were also considered that could cause such antibodies. In addition, the assumption was examined as to whether or not the loss of suppressor cells correlates with the presence of autoantibodies. The incidence of autoantibodies mentioned was not found to be statistically higher in our study; most of the autoantibodies could be explained by other accompanying diseases. Only in the T-suppressor cells was there a significant decrease. Patients with autoantibodies showed no more deficit in T-suppressor cells than patients without autoantibodies. PMID- 2965692 TI - Mediation of in vivo tumor-neutralizing activity by Lyt-2+ as well as L3T4+ T cell subsets. AB - The present study reexamines the cell surface nature of T cells mediating in vivo protective tumor immunity with the use of anti-L3T4 and -Lyt-2 antibodies. C3H/HeN mice hyperimmune against syngeneic MH134 hepatoma or MCH-l-Al fibrosarcoma were prepared by intradermal (id) inoculation of viable tumor cells followed by surgical resection of the tumor and by repeated challenges with viable tumor cells. Spleen cells from these mice were fractionated into L3T4+ or Lyt-2+ T cell subset by treatment with anti-Lyt-2 or -L3T4 antibody plus complement (C). Winn assays performed by utilizing such fractionated T cells have revealed that both L3T4+ and Lyt-2+ T cell subsets from hyperimmune mice produced complete tumor protection. Flow microfluorometry study illustrated that the treatment with anti-L3T4 or -Lyt-2 antibody plus C resulted in the complete isolation of L3T4- Lyt-2+ (Lyt-2+) or L3T4+ Lyt-2- (L3T4+) T cell subset, respectively. This contrasted with the failure of treatment with anti-Lyt-1 antibody plus C to isolate all T cells expressing Lyt-2 marker. It was further demonstrated that each subset of T cells exerted its anti-tumor effect in a tumor specific way and without a requirement for the other alternative subpopulation of unprimed T cells. These results indicate that Lyt-2+ T cell subset can be successfully isolated by treatment with anti-L3T4 but not with anti-Lyt-1 antibody plus C, and that each single subset of Lyt-2+ and L3T4+ T cells can function as in vivo effector T cells. PMID- 2965694 TI - Protease-inhibitory activities of leupeptin analogues. AB - Thirty analogues of leupeptin were synthesized and examined for their inhibitory activities against trypsin, papain, plasmin, kallikrein, thrombin and urokinase in vitro. Benzoyl- and alpha-naphthalenesulfonyl-L-leucyl-L-argininal were 8 times more inhibitory to papain, benzyloxycarbonyl-L-pyroglutamyl-L-leucyl-L argininal 10 times more to trypsin and plasmin, and DL-2-pipecolyl-L-leucyl-L argininal 25 times more to kallikrein than leupeptin. Against urokinase, only L pyroglutamyl-L-leucyl-L-argininal exhibited a potent inhibitory activity. alpha Naphthalensulfonyl-, dansyl- and benzyloxycarbonyl-(2S,3R)-3-amino-2-hydroxy-4 phenylbutyryl-L-leucyl-L- argininal were inhibitory to thrombin. PMID- 2965693 TI - Muscle fibre types in the external eye muscles of the pigeon, Columba livia. AB - Fibre typing with antisera raised against specific myosin types from muscles of known physiological properties were used to characterise the fibre types within the oculorotatory muscles of pigeons. Fibres reacting strongly to antiserum anti ALD (specific for tonic fibre myosin) were found lying along the global margin of the muscle and also in a layer lying immediately beneath a discrete band of fibres running along the orbital margin. These fibres resembled those of the skeletal muscle ALD in their type properties. Using another antiserum, anti-I, specific for slow twitch and to a lesser extent, slow tonic myosins, it was possible to identify another slow fibre type which formed the orbital layer and also lay scattered randomly through the body of the muscle. No equivalent to this type was found in the skeletal muscles ALD or iliofibularis. The remaining fibres which did not react with either anti-ALD or anti-I formed 58% of the fibre population and reacted with an antiserum specific for fast myosin. However, their response to alkali preincubation suggests that the fast fibres of eye muscles also contain a myosin which is different from those in skeletal muscle. PMID- 2965695 TI - Effects of metabolic blockade on distribution of blood flow to respiratory muscles. AB - Sublethal inhibition of citrate metabolism in the tricarboxylic acid (TCA) cycle with monofluoroacetate (MFA) has been shown to cause a fivefold increase in myocardial blood flow without any change in cardiac output, blood pressure, or O2 consumption (C. Liang, J. Clin. Invest. 60: 61-69, 1977); however, blood flow did not increase to any organs examined other than the heart, including resting limb skeletal muscle. Preferential inhibition of glycolysis with iodoacetate (IA) failed to cause similar changes in distribution of blood flow. This unique response of myocardium to TCA cycle inhibition suggested a unique metabolic control of cardiac vasodilation. An alternate explanation is that MFA is preferentially concentrated in active muscle. After MFA, tissue citrate accumulates behind the block and the highest levels are reported in the heart and diaphragm, suggesting enhanced blockade or enhanced compensation in these two continuously active muscles. To test the hypothesis that vasodilation in the heart after MFA is not unique and that similar vasodilation will be evoked in active respiratory muscles, we measured blood flow to the myocardium, kidney, diaphragm, intercostals, transverse abdominals, and triceps brachii in anesthetized dogs using radionuclide-labeled microspheres, before and after MFA, and in another set of dogs before and after IA. Before MFA or IA, inspiratory loading significantly increased blood flow to active muscles of breathing in proportion to the added load. After MFA, blood flow to active muscles of breathing as well as to the heart became abnormally elevated with respect to mechanical work, and loading evoked no further increase in blood flow.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965696 TI - The triangle--medical therapy, PTCA, and by-pass surgery for coronary artery disease. PMID- 2965697 TI - Two pathways for oxygen exchange by heavy meromyosin and their dependence on actin. AB - In the hydrolysis of MgATP by acto heavy meromyosin (HMM) there are two enzymatic pathways that differ in the properties of their intermediate oxygen exchange; one of these is designated the low exchange pathway (P1); the other is designated the high exchange pathway (P2). A plot of the P1 flux versus the actin concentration gives a sigmoid curve, whereas the corresponding curve for the P2 flux rises in an approximately hyperbolic manner. At low concentrations of actin, where the sigmoid curve of the P1 flux is in a lag phase, the major flux is along P2; but at higher concentrations of actin, as the P1 curve rises sharply, the flux along P1 comes to predominate. Even at the highest levels of actin, at saturating levels for both pathways, the kinetics of exchange along P1 and P2 are significantly different. In addition to these differences in the actin dependence, the flux of P1 relative to P2 is markedly inhibited by KCl. Therefore, which of the two pathways dominates during the hydrolysis of MgATP by HMM is strongly dependent on experimental conditions. The findings suggest that P1 involves the interaction of HMM with two actin units whereas P2 involves the interaction of HMM with one actin unit. The results are discussed in relation to a kinetic scheme based on this proposal. PMID- 2965698 TI - Interaction of human monomeric C3b with its receptor (complement receptor type 1, CR1) on neutrophils. Evidence for negative cooperativity. AB - The binding of highly purified monomeric 125I-C3b to its receptor (CR1) on resting human polymorphonuclear neutrophils (PMN) was analyzed under equilibrium conditions, at 4 degrees C and low ionic strength. Scatchard analysis of specific binding data yielded curvilinear concave upward plots, which resulted from the presence of site-site interactions of the negative type among PMN C3b-receptors (negative cooperativity), as shown by dissociation kinetic experiments. Indeed, the dissociation rate of 125I-C3b from PMN was markedly increased in the presence of an excess of unlabeled C3b in the dilution medium and was directly dependent on the degree of initial receptor occupancy with the radioligand. These interactions occurred when 2% of the receptors were occupied with 125I-C3b and resulted in a 4-fold decrease in CR1 affinity when the receptor went from its "empty" to its "filled" conformation. In a disease associated with a continuous production of C3b (factor I deficiency), CR1 on in vivo circulating PMN was found to be in a "low affinity" and "high dissociating" state similar to that of normal CR1 at high occupancy. Finally, negative cooperativity among CR1 sites disappeared after PMN activation with chemotactic peptides. PMID- 2965699 TI - The structure of the amino terminus of tropomyosin is critical for binding to actin in the absence and presence of troponin. AB - Analysis of two recombinant variants of chicken striated muscle alpha-tropomyosin has shown that the structure of the amino terminus is crucial for most aspects of tropomyosin function: affinity to actin, promotion of binding to actin by troponin, and regulation of the actomyosin MgATPase. Initial characterization of variants expressed and isolated from Escherichia coli has been published (Hitchcock-DeGregori, S. E., and Heald, R. W. (1987) J. Biol. Chem. 262, 9730 9735). Fusion tropomyosin contains 80 amino acids of a nonstructural influenza virus protein (NS1) on the amino terminus. Nonfusion tropomyosin is a variant because the amino-terminal methionine is not acetylated (unacetylated tropomyosin). The affinity of tropomyosin labeled at Cys190 with N [14C]ethylmaleimide for actin was measured by cosedimentation in a Beckman Airfuge. Fusion tropomyosin binds to actin with an affinity slightly greater than that of chicken striated muscle alpha-tropomyosin (Kapp = 1-2 X 10(7) versus 0.5 1 X 10(7) M-1) and more strongly than unacetylated tropomyosin (Kapp = 3 X 10(5) M-1). Both variants bind cooperatively to actin. Troponin increases the affinity of unacetylated tropomyosin for actin (+Ca2+, Kapp = 6 X 10(6) M-1; +EGTA, Kapp = 2 X 10(7) M-1), but the affinity is still lower than that of muscle tropomyosin for actin in the presence of troponin (Kapp much greater than 10(8) M-1). Troponin has no effect on the affinity of fusion tropomyosin for actin indicating that binding of troponin T to the over-lap region of the adjacent tropomyosin, presumably sterically prevented by the fusion peptide in fusion tropomyosin, is required for troponin to promote the binding of tropomyosin to actin. The role of troponin T in regulation and the mechanisms of cooperative binding of tropomyosin to actin have been discussed in relation to this work. PMID- 2965700 TI - Electron microscope observations on Ca2+-ATPase microcrystals in detergent solubilized sarcoplasmic reticulum. AB - Crystalline arrays of Ca2+-ATPase molecules develop in detergent-solubilized sarcoplasmic reticulum during incubation for several weeks at 2 degrees C under nitrogen in a medium of 0.1 M KCl, 10 mM K-3-(N-morpholino)propanesulfonate, pH 6.0, 3 mM MgCl2, 20 mM CaCl2, 20% glycerol, 3 mM NaN3, 5 mM dithiothreitol, 25 IU/ml Trasylol, 2 micrograms/ml 1,6-di-tert-butyl-p-cresol, 2 mg/ml protein, and 2-4 mg of detergent/mg of protein. Electron microscopy of sectioned, negatively stained, freeze-fractured, and frozen-hydrated Ca2+-ATPase crystals indicates that they consist of stacked lamellar arrays of Ca2+-ATPase molecules. Prominent periodicities of ATPase molecules within the lamellae arise from a centered rectangular lattice of dimensions 164 x 55.5 A. The association of lamellae into three-dimensional stacks is assumed to involve interactions between the exposed hydrophilic headgroups of ATPase molecules, that is promoted by glycerol and 20 mM Ca2+. Similar Ca2+-induced crystals were observed with purified or purified and delipidated Ca2+-ATPase preparations at lower detergent/protein ratios. Cross linking of Ca2+-ATPase crystals with glutaraldehyde protects the structure against conditions such as low Ca2+, high pH, elevated temperature, SH group reagents, high concentration of detergents, and removal of phospholipids by extraction with organic solvents that disrupt unfixed preparations. PMID- 2965701 TI - The purification and characterization of mannosidase IA from rat liver Golgi membranes. AB - Rat liver Golgi membranes contain two alpha 1,2-specific mannosidases (IA and IB) (Tulsiani, D. R. P., Hubbard, S. C., Robbins, P. W., and Touster, O. (1982) J. Biol. Chem. 257, 3660-3668). Mannosidase IA has now been purified to apparent homogeneity by detergent extraction and (NH4)2SO4 precipitation, followed by Sephacryl S-300, ion-exchange, and hydroxylapatite chromatography. The enzyme was homogeneous by nondenaturing polyacrylamide gel electrophoresis with different gel concentrations, and Ferguson plot analysis indicated an Mr of 230,000 for the native enzyme. Although electrophoresis under denaturing conditions generally gave a subunit Mr of 57,000, electrophoresis of less than 1 microgram of protein yielded a faint doublet of Mr 57,000 and 58,000. Thus, the enzyme appears to be a tetramer with four very similar subunits. The enzyme bound to concanavalin A Sepharose 4B only when it was kept in contact with the lectin for 16 h. Endoglycosidase H treatment resulted in loss of its binding to the lectin, without leading to a detectable change in the size of the enzyme subunit. On electrophoretic gels, the enzyme gave a faint positive stain with periodic acid Schiff's base. The enzyme contained about 0.9% hexose by direct analysis. It did not bind to affinity resins specific for neuraminic acid, galactose, or N acetylglucosamine. All these studies suggest that the enzyme is a glycoprotein containing only one or two clusters of high mannose oligosaccharide. Mannosidase IA is active toward oligosaccharides containing alpha 1,2-linked mannosyl residues. [3H]Man9GlcNAc, [3H] Man8GlcNAc, [3H]Man7GlcNAc, and [3H]Man6GlcNAc are good substrates. Man9GlcNAc, the best substrate, yields Man8, Man7, and Man6 derivatives with structures suggesting that the sequence of release of mannose residues is rather specific. Immunoprecipitation studies using polyclonal antibody (IgG) prepared against homogeneous mannosidase IA cross-reacted with mannosidase IB, a result suggesting that these two enzymes share antigenic determinants. However, no cross-reactivity was observed with rat liver cytosolic and lysosomal alpha-D-mannosidases or with Golgi mannosidase II. PMID- 2965702 TI - Effects of inhibitors of N-linked oligosaccharide processing on the biosynthesis and function of insulin and insulin-like growth factor-I receptors. AB - We have used specific inhibitors of oligosaccharide processing enzymes as probes to determine the involvement of oligosaccharide residues in the biosynthesis and function of insulin and insulin-like growth factor-I receptors. In a previous study (Duronio, V., Jacobs, S., and Cuatrecasas, P. (1986) J. Biol. Chem. 261, 970-975) swainsonine was used to inhibit mannosidase II, resulting in the production of receptors containing only hybrid-type oligosaccharides. These receptors had a slightly lower molecular weight and were much more sensitive to endoglycosidase H, but otherwise behaved identically to normal receptors. In this study, we used two compounds that inhibit oligosaccharide processing at earlier steps: (i) N-methyl-1-deoxynojirimycin (MedJN), which inhibits glucosidases I and II and yields glucosylated, high mannose oligosaccharides, and (ii) manno-1 deoxynojirimycin (MandJN), which inhibits mannosidase I and yields high mannose oligosaccharides. In the presence of MandJN, HepG2 cells synthesized receptors of lower molecular weight, which were cleaved into alpha and beta subunits and were able to bind hormone and autophosphorylate. These receptors were as sensitive to endoglycosidase H as receptors made in the presence of swainsonine. In the presence of MedJN, receptors of only slightly lower molecular weight than normal were synthesized and were shown to contain some glucosylated high mannose oligosaccharides. These receptors were able to bind hormone and retained hormone sensitive autophosphorylation activity. In both cases, the incompletely processed receptors could be detected at the cell surface by cross-linking of iodinated hormone and susceptibility to trypsin digestion, although less receptor was present in cells treated with MedJN. Studies of receptor synthesis using pulse chase labeling showed that the receptor precursors synthesized in the presence of MedJN were cleaved into alpha and beta subunits at a slower rate than normal receptors or those made in the presence of MandJN. Inhibition of oligosaccharide processing had no effect on the association of the receptor subunits into disulfide-linked oligomeric complexes. PMID- 2965703 TI - Changes in the ATPase activity of insect fibrillar flight muscle during sinusoidal length oscillation probed by phosphate-water oxygen exchange. AB - Extensive phosphate-water oxygen exchange occurs when ATP is hydrolyzed in an [18O]water medium by length oscillated and Ca2+-activated, chemically skinned fibers from the flight muscle of the giant waterbug Lethocerus indicus. For fibers which are length oscillated under conditions not optimal for ATPase activity or oscillatory work, the pattern of exchange shows two pathways for hydrolysis. One pathway has low exchange, because steps controlling Pi release are rapid; the other pathway has high exchange and slow Pi release. Steps controlling Pi release appear rate-limiting for changes in the high-exchange ATPase activity that occur on varying the frequency and amplitude of oscillation. On length oscillation under conditions of optimal ATPase activity or work, only the high-exchange pathway is present. Cross-bridges following the high-exchange pathway are therefore responsible for oscillatory work, the physiological function of the muscle, and behave uniformly with respect to oxygen exchange. The single pathway and the magnitude of the ATPase activity are both similar to results with isometric strained fibers (Lund, J., Webb, M. R., and White, D. C. S. (1987) J. Biol. Chem. 262, 8584-8590). A qualitative model is suggested for oscillatory work by cross-bridges, arising from the common periodicity of the thick and thin filaments in insect flight muscle. PMID- 2965704 TI - Biochemical and thrombolytic properties of a low molecular weight form (comprising Leu144 through Leu411) of recombinant single-chain urokinase-type plasminogen activator. AB - The cDNA encoding a low Mr derivative (residues 144-411) of human single-chain urokinase-type plasminogen activator was cloned, the recombinant low Mr single chain urokinase-type plasminogen activator (rscu-PA-32k) was expressed in Chinese hamster ovary cells, and the translation product was purified to homogeneity from conditioned cell culture medium. rscu-PA-32k is very similar to intact recombinant single-chain urokinase-type plasminogen activator in terms of its very low activity (120 IU/mg) on a chromogenic substrate for urokinase (pyroglutamylglycylarginine p-nitroanilide), its plasminogen-dependent fibrinolytic activity on fibrin plates (specific activity = 170,000 IU/mg), its plasminogen activating potential, and the lack of specific binding to fibrin. In a rabbit jugular vein thrombosis model, comparable thrombolysis was obtained with rscu-PA-32k as compared to low molecular weight two-chain urokinase (50% lysis at 2.1 and 1.6 mg/kg infused over 4 h). Thrombolysis was associated with much less extensive systemic fibrinogen breakdown with rscu-PA-32k than with two-chain urokinase (residual fibrinogen at 50% lysis of 71 and 10%, respectively). It is concluded that the functional properties of rscu-PA-32k, expressed with a high efficiency, are similar to those of its previously characterized natural counterpart. PMID- 2965706 TI - Expression and dexamethasone regulation of the human corticotropin-releasing hormone gene in a mouse anterior pituitary cell line. AB - The factors controlling the expression of corticotropin-releasing hormone (CRH), a hypothalamic neuropeptide involved in the regulation of ACTH secretion, are poorly understood partly because a suitable in vitro model is lacking. To study the regulation of CRH gene expression, an 8-kilobase (kb) DNA fragment containing the entire human CRH gene as well as approximately 6 kb of 5' sequence and 0.8 kb of 3' sequence was isolated from a lambda Charon 4A human genomic library and introduced into a mouse anterior pituitary cell line, AtT-20, by CaPO4 transfection with a neomycin-selectable marker. Approximately 10% of the neomycin resistant lines stably expressed the CRH gene and secreted radioimmunoassay detectable CRH into culture media at levels greater than 100 pg/ml. By Southern blot analysis the 8-kb DNA fragment containing the CRH gene had been incorporated intact into the AtT-20 genome. In each CRH-producing strain, but not in the parent AtT-20 cell line, we detected by Northern blot analysis an RNA species that hybridized to two radioactive cRNA probes specific for either the 5' or 3' portion of CRH mRNA, and that co-migrated with placental CRH mRNA. Dexamethasone treatment for 24-96 h caused a specific decrease in CRH mRNA and peptide levels of 40-50% in the five CRH-producing cell lines with half-maximal suppression at approximately 10(-9) M dexamethasone, indicating that CRH gene expression is negatively regulated by glucocorticoids. Thus, we have established an in vitro model suitable for studying in detail those cis- and trans-acting factors which regulate CRH gene expression. PMID- 2965705 TI - The effect of the protein matrix on glycan processing in glycoproteins. Kinetic analysis of three rat liver Golgi enzymes. AB - In order to assess the basis for the regulatory effects of the protein matrix on the processing of glycans in glycoproteins, we have used the avidin biotinylglycan neoglycoprotein model system to compare the kinetic parameters for three rat liver Golgi enzymes acting on their free and protein-bound glycan substrates. Two modes of glycan display in the avidin complex were produced by the use of either the biotinyl- or the 6-biotinamidohexanoyl-group as ligands for the avidin binding. N-Acetylglucosaminyltransferase I gave a 100-fold decrease in Vmax/Km for the avidin complex of Man5GlcNAc2-(biotinyl)Asn as compared to the free glycan derivative; the rate difference reflects a large (25x) decrease in the Vmax and a relatively small increase (4x) in Km. When the substrate with the extension arm (Man5GlcNAc2-(6-biotinamidohexanoyl)Asn) was used, the difference between Vmax/Km for free and avidin-bound substrate was only 6-fold. The Vmax/Km ratio for N-acetylglucosaminyltransferase II also showed a 10-fold difference for free and avidin-bound GlcNAcMan3GlcNAc2-(biotinyl)Asn; the introduction of the extension arm in the complex reduced the difference to about 3-fold. The third enzyme, galactosyltransferase, acting on the substrate GlcNAcMan5-GlcNAc2-R in the presence of the mannosidase II-inhibitor swainsonine, showed a small, 2- to 3 fold, decrease in the Vmax for the bound substrates, both with and without the extension arm. The results suggest that the protein matrix affects the catalytic efficiency rather than the substrate affinity of the processing enzymes. PMID- 2965707 TI - Laser Doppler monitoring of microcirculatory changes in acute burn wounds. AB - Because burns are dynamic wounds that can change in apparent depth during the first 72 hours, we asked whether measuring changes in cutaneous blood flow might help predict the ultimate fate of burns that were not obviously shallow or deep. A laser Doppler flowmeter was used to study cutaneous perfusion for at least 72 hours in partial-thickness wounds on patients with burns of less than 15% TBSA and in experimental wounds of similar size on rats. Clinical wounds that healed without grafting consistently showed elevated perfusion levels which increased further within 72 hours, whereas wounds eventually requiring grafting experienced lower perfusion levels with no obvious pattern of increase. Differences between average flow levels for healing and nonhealing burns were statistically significant throughout the study period. Perfusion levels in experimental wounds were stratified according to burn severity, with shallower wounds showing a pattern of increase similar to the clinical wounds. PMID- 2965708 TI - Pulse oximetry for vascular monitoring in burned upper extremities. AB - The reliability and accuracy of pulse oximetry as a monitoring device in deep superficial and full-thickness circumferential burns of the upper extremity were evaluated clinically. Pulse oximetry was correlated with the laser Doppler flowmeter experimentally in healthy volunteers. By continuously monitoring oxygen saturation levels, the pulse oximeter detected extremity hypoxia before it resulted in an adverse outcome. Extremities exhibiting oxygen saturation above 95% were treated with early excision and skin grafting without escharotomy. Extremities with O2 saturations of less than 95% underwent immediate escharotomy with return to normal saturation levels. The pulse oximeter is easy to use by nursing personnel; is noninvasive and accurate; and displays the oxygen saturation level within seconds. PMID- 2965709 TI - Pro-opiomelanocortin peptides in reproductive physiology. PMID- 2965710 TI - Nerve blocks for pain management. PMID- 2965711 TI - Laparoscopic control of danazol therapy on pelvic endometriosis. AB - In order to evaluate the efficiency of Danazol treatment on pelvic endometriosis, a laparoscopic control with photographs was performed before treatment in 33 cases. Of these 20 were severe (group A), seven mild (group B) and six light (group C), according to the American Fertility Society classification, and 15 patients were infertile, 10 had pelvic pain and eight had dysmenorrhoea. Among the 28 patients who accepted a laparoscopic control after 6 months of treatment, the result was quite good in 50% of the cases in groups B and C, an improvement was seen in 30% of the patients in groups A and B, but a failure in 50% of the cases, especially in group A. Of the 14 cases with an ovarian cyst greater than 3 cm in diameter an operation was necessary either by laparoscopy (four cases) or by laparotomy (five cases). Seven pregnancies were established out of 15 cases of infertility, but a relapse was noticed in 30% of the cases. In conclusion, it appears that Danazol therapy is effective in mild or light endometriosis, but is not sufficient in cases of ovarian endometriosis with cysts greater than 3 cm in diameter. PMID- 2965712 TI - Ovarian stimulation for in-vitro fertilization combining administration of gonadotrophins and blockade of the pituitary with D-Trp6-LHRH microcapsules: pilot studies with two protocols. AB - In women undergoing in-vitro fertilization and embryo transfer (IVF-ET), a total of 408 IVF cycles were stimulated using human menopausal gonadotrophin (HMG) or pure follicle stimulating hormone (FSH) plus HMG in combination with a single injection of D-Trp6-LHRH microcapsules in order to enhance the ovarian response to gonadotrophins and to avoid spontaneous LH surges. Sixty-seven pregnancies were achieved. Two protocols were employed. In protocol 1 ('blocking protocol', n = 268), the pituitary was first inhibited with a full dose (3.75 mg) of D-Trp6 LHRH in microcapsules and ovarian stimulation was started after the hypogonadotrophic hypogonadal state was ascertained (E2 less than 50 pg/ml). In protocol 2 ('flare-up protocol', n = 140), the treatment with D-Trp6-LHRH microcapsules (half-dose = 1.80 mg) and the ovarian stimulation with gonadotrophins were started at the same time. Higher doses of gonadotrophins were needed (39.5 +/- 11.2 ampoules FSH and/or HMG) in protocol 1, in which the pituitary was blocked prior to and during the stimulation, than in protocol 2 (20 +/- 9 ampoules) where the exogenous gonadotrophin stimulation appeared to be augmented by the initial agonistic effect of the injection of D-Trp6-LHRH microcapsules. In patients with purely tubal infertility, under 38 years old and no male factor, the results obtained with protocols 1 and 2 were similar in terms of pregnancy rate per cycle or per embryo transfer: 22.6 versus 20.5% and 28.3 versus 27.4%, respectively. However, considering the cost benefit, 'flare-up' protocols appeared to be a better choice and could be recommended. PMID- 2965713 TI - Lack of endometriosis in patients with repeated abortion. AB - Laparoscopy was performed in 25 consecutive patients with two or more (range 2-6, mean 2.8) histologically documented first-trimester abortions in whom the well established causes for recurrent miscarriages had been excluded. Endometriosis could not be diagnosed in any of the 25 women. Thus our results are against endometriosis as a cause of repeated abortion and do not support the routine inclusion of laparoscopy in the evaluation of the patient with recurrent abortions. PMID- 2965714 TI - Rotational endarterectomy in normal canine coronary arteries: preliminary report. AB - Endarterectomy was performed in vivo using a high speed rotating abrasive-tipped catheter device in 11 normal canine coronary arteries. The device is designed to remove atheromatous material from diseased arteries by the abrasive action of its rotating tip. It was operated percutaneously from a femoral approach using conventional angioplasty guiding equipment. The rotating device was advanced over a guide wire from just beyond the tip of the guide catheter into the distal vessel. Six arteries were harvested immediately after endarterectomy and five were left in place for 7 +/- 2.8 days; in the latter group, the animals were maintained on a regimen of aspirin, 325 mg/day. Angiography before and after treatment demonstrated vessel patency in all cases. Caliper-measured luminal diameters were not significantly changed after endarterectomy. Histologic examination of pressure-fixed vessels showed extensive intimal loss and 20 to 30% loss of the internal elastic lamina. Medial damage was superficial and never exceeded 40% of the total medial thickness. There were no vessel perforations. Results of histologic study of the myocardium supplied by the treated vessels were normal without evidence of distal embolization or infarction. It is concluded that a high speed rotating abrasive device can be safely operated percutaneously in normal coronary arteries and results in minimal vessel damage and continued patency at 7 +/- 2.8 days. PMID- 2965715 TI - Compromise of beneficial effects of reperfusion on myocardium supplied by vessels with critical residual stenosis. AB - Coronary thrombolysis in patients frequently unmasks high grade residual stenosis. To determine whether beneficial effects of reperfusion are compromised by critical residual coronary stenosis, 14 dogs were instrumented with an external left anterior descending coronary artery balloon occluder, Doppler flow probe and adjustable screw clamp. In eight of the dogs, critical stenosis (abolition of reactive hyperemia after a 20 s occlusion; 95.7 +/- 1.0% cross sectional area reduction) was induced before occlusion and maintained. In the control group (n = 6), no stenosis was induced. Each dog was subjected to 2 h of myocardial ischemia followed by balloon deflation and 24 h of reperfusion. Myocardial blood flow assessed with microspheres was similar during balloon inflation in both groups and indicative of profound ischemia. Transmural blood flow to the reperfused zone assessed 1 min after balloon deflation was significantly greater in control dogs without residual stenosis (383% of normal compared with 120% of normal in dogs with stenosis) (p less than 0.01). Compromise of transmural flow persisted in dogs with stenosis (85% compared with 121% of normal in control dogs after 1 h, p less than 0.05; and 49% compared with 68% after 24 h of reperfusion, p less than 0.05). Diminution of subendocardial blood flow after reperfusion was particularly marked. The extent of infarction was greater in the heart of dogs with residual stenosis. Thus, residual critical coronary stenosis compromises nutritional perfusion and salvage of reperfused myocardium after recanalization. These observations underscore the need for prompt identification of patients with high grade residual stenosis early after coronary thrombolysis and the potential value of angioplasty or coronary surgery in selected patients soon after initial recanalization. PMID- 2965716 TI - Emergency coronary artery bypass surgery preserves global and regional left ventricular function after intravenous tissue plasminogen activator therapy for acute myocardial infarction. AB - Emergency coronary bypass surgery was performed in 24 (6.2%) of 386 consecutive patients enrolled in the Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) Multicenter Trial. Intravenous tissue plasminogen activator was administered 2.6 +/- 0.7 h and bypass surgery was performed 7.3 +/- 1.9 h after the onset of infarction. Infarct artery patency was achieved in 21 (88%) of the 24 patients (pharmacologically in 18 or mechanically with coronary angioplasty in 3) in the catheterization laboratory before bypass surgery. The indication for surgery was left main or equivalent coronary artery disease in 7 patients, coronary anatomy unsuitable for angioplasty in 4 patients and unsuccessful coronary angioplasty in 13 patients. A coronary perfusion catheter was inserted before surgery in 11 of 13 patients with unsuccessful angioplasty. All three deaths occurred postoperatively in patients with preoperative cardiogenic shock. Three patients required surgical reexploration for postoperative hemorrhage. Comparison of preoperative and predischarge contrast left ventriculograms demonstrated significant preservation of global (left ventricular ejection fraction 49 +/- 6 to 56 +/- 6%; p = 0.008) and regional (standard deviation/chord -2.6 +/- 0.5 to -1.5 +/- 1.1; p = 0.001) left ventricular function. Emergency coronary bypass surgery can be performed with a low morbidity and mortality in patients treated with intravenous tissue plasminogen activator therapy for acute myocardial infarction. Such therapy is associated with significant preservation of global and regional (infarct zone) left ventricular function. PMID- 2965717 TI - Clinical significance of perfusion defects by thallium-201 single photon emission tomography following oral dipyridamole early after coronary angioplasty. AB - The clinical significance of myocardial perfusion defects present early after angiographically successful percutaneous transluminal coronary angioplasty was assessed in 53 patients using thallium-201 single photon emission computed tomography combined with pharmacologic vasodilation induced by a large dose (300 mg) of orally administered dipyridamole. Myocardial tomographic images were obtained at a mean of 20 +/- 6 h (SD) before and 2.9 +/- 2.7 days after angioplasty. Before angioplasty, 15 (28%) of the 53 patients developed angina after dipyridamole administration, in contrast to only 3 (7.5%) of 40 patients after angioplasty (p less than 0.001). The mean percent luminal area stenosis decreased from 93 +/- 6% before angioplasty to 34 +/- 17% after angioplasty (p less than 0.001). Myocardial perfusion defects, present in 49 (93%) of the 53 patients before angioplasty, were reversible in 44 patients (83%), all of whom underwent dilation of arteries supplying the ischemic areas. After angioplasty, 26 (65%) of 40 patients had no ischemic defects, whereas 14 (35%) of the patients still had an ischemic defect in the vascular territory of the dilated artery. After a mean follow-up period of 21.7 months, 13 (33%) of 39 patients developed restenosis, 10 of whom had an ischemic defect early after angioplasty. Restenosis developed in 10 (71%) of 14 patients with an ischemic defect after angioplasty, but in only 3 (11.5%) of the patients without an ischemic defect (p = 0.007). In conclusion, thallium-201 tomography after oral dipyridamole affords convenient assessment of the physiologic significance of coronary stenosis present before angioplasty and the residual stenosis after angioplasty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965718 TI - Transluminal intracoronary reperfusion catheter: a device to maintain coronary perfusion between failed coronary angioplasty and emergency coronary bypass surgery. AB - The reperfusion catheter is a 4.3F catheter with 30 holes over its distal 10 cm. It is used to maintain coronary blood flow in patients awaiting emergency coronary bypass surgery after failed coronary angioplasty. The insertion of the reperfusion catheter was attempted in 20 patients (14 with total occlusion and 6 with severe residual stenosis judged to be in jeopardy of reclosure before operation). The reperfusion catheter was successfully placed across the obstruction in 18 patients (90%). After successful insertion of the reperfusion catheter, 16 patients had good anterograde flow (Thrombolysis in Myocardial Infaction [TIMI] trial grade II or III); angiographic improvement was associated with significant lessening of ST segment elevation as well as a decrease in chest pain in most patients. Two patients had poor or absent anterograde flow (TIMI grade O or I) because of extensive preexisting intracoronary thrombosis; one died from refractory ventricular fibrillation. In each of the remaining patients emergency coronary bypass surgery was performed with no deaths or significant cardiac complications. The reperfusion catheter is a safe and effective method to reestablish and maintain coronary blood flow before coronary bypass surgery after failed coronary angioplasty. Because there is the potential risk of serious complications, particularly thrombus formation within this catheter, the reperfusion catheter should be used cautiously and the patient should undergo immediate bypass surgery. PMID- 2965719 TI - Double-blind placebo-controlled study of loratadine, mequitazine, and placebo in the symptomatic treatment of seasonal allergic rhinitis. AB - Loratadine is a long-acting H1 antagonist devoid of anticholinergic and sedative effects. A double-blind, placebo-controlled, parallel-group study was performed in 69 patients to compare efficacy and safety of loratadine and mequitazine. Patients allergic to grass pollens were randomly assigned to one of the three treatment groups and followed up to 2 weeks during the peak of the pollen season. Symptoms of allergic rhinitis were evaluated at baseline and after 3, 7, and 14 days of treatment by the physician with patients rating their response daily on diary cards. Both loratadine and mequitazine induced a significant relief of nasal symptoms when these were compared to placebo. Loratadine was found to be significantly superior to placebo after 3 days of treatment, whereas a significant improvement was only observed after 7 days in patients treated with mequitazine. For nonnasal symptoms, none of the two anti-H1 antagonist induced a significant improvement, and this lack of effect may be related to low symptoms at baseline. Loratadine did not induce more side effects than placebo. Loratadine can be considered to be an effective and safe anti H1 histamine with a rapid onset of action. PMID- 2965720 TI - Mapping gold-labeled IgE receptors on mast cells by scanning electron microscopy: receptor distributions revealed by silver enhancement, backscattered electron imaging, and digital image analysis. AB - Immunogold labeling and silver enhancement techniques are widely used to determine density and distribution of cell membrane receptors by light and transmission electron microscopy. However, these techniques have not been widely used for receptor detection by scanning electron microscopy. We used antigen- or protein A-conjugated colloidal gold particles, together with silver enhancement, sequential secondary and back-scattered electron imaging (SEI and BEI), and digital image processing, to explore cell surface distribution of IgE-receptor complexes on RBL-2H3 cells, a rat leukemia line that provides a model for the study of mucosal mast cells. Cells were first incubated with a monoclonal antidinitrophenol IgE (anti-DNP-IgE) that binds with high affinity to cell surface IgE receptors. The resulting IgE-receptor complexes were cross-linked either with the multivalent antigen, DNP-BSA-gold, or with a polyclonal anti-IgE antibody. Antibody-treated cells were labeled after fixation with protein A-gold. Fixed, gold-labeled cell monolayers were silver enhanced (or not), dehydrated, critical point-dried, and coated with gold-palladium (for SEI analysis) or carbon (for combined SEI/BEI analysis). They were observed in an Hitachi S800 SEM equipped with a field emission tip and a Robinson backscattered electron detector. An image processor (MegaVision 1024XM) digitized images directly from the S800 microscope at 500-1000 line resolution. Silver enhancement significantly improves detection of gold particles in both SEI and BEI modes of SEM. On gold palladium-coated samples, 20-nm particles are resolved by SEI after enhancement. BEI resolves 15-nm particles without enhancement and 5- or 10-nm particles are resolved by BEI on silver-enhanced, carbon-coated samples. Neither BEI nor SEI alone can yield high resolution topographical maps of receptor distribution (BEI forms images on the basis of atomic number contrast which reveals gold but not surface features). Image analysis techniques were therefore introduced to digitize, enhance, and process BEI and SEI images of the same field of view. The resulting high-contrast, high-resolution images were superimposed, yielding well resolved maps of the distribution of antigen-IgE-receptor complexes on the surface of RBL-2H3 mast cells. The maps are stored in digital form, as required for computer-based quantitative morphometric analyses. These techniques of silver enhancement, combined BEI/SEI imaging, and digital image analysis can be applied to analyze density and distribution of any gold-labeled ligand on its target cell. PMID- 2965721 TI - The development of helper T cell precursors in mouse thymus. AB - We have examined the appearance in mouse ontogeny of thymocyte precursors for Ag specific, MHC-restricted Th. These cells are first detectable at day 18 of fetal life, about 1 day after alpha/beta, TCR-positive cells begin to appear. These early Th precursors are not dependent on the thymus for priming with Ag and MHC, and are L3T4+, Lyt-2-. Thus, these cells already have the phenotype of mature Th. In neonatal F1 animals expressing both IAk and IAb, the appearance of Th precursors restricted by either IAk or IAb is specifically inhibited by treatment of the mice with anti-IAk or anti-IAb antibodies, respectively. These results indicate that cells of mature T cell phenotype and function can arise fairly rapidly from immature, receptor-bearing precursors, once these appear. Moreover the results are in line with those previously obtained in chimeric animal experiments which suggested that specific interaction of TCR on thymocytes with class II alleles in the thymus is required for the subsequent appearance of T cells restricted by those class II alleles. PMID- 2965722 TI - IL-2-dependent expansion of CD3+ large granular lymphocytes expressing T cell receptor-gamma delta. Evidence for a functional receptor by anti-CD3 activation of cytolysis. AB - The nature and function of the TCR on PBL of a patient with a chronic CD3+ large granular (LGL) proliferation was studied. Fresh peripheral blood from this individual was comprised of 80% lymphocytes, 65 to 75% of which were CD3+, CD8+, Leu-7+ LGL. Of these LGL, 72% initially expressed the TCR-alpha beta heterodimer, whereas 21% did not. Cytotoxicity directed against MHC-unrestricted targets was minimal. After several days of exposure to rIL-2, cytotoxic activity was greatly enhanced, correlating with a disappearance of CD3+ cells expressing the alpha beta heterodimer. Twelve days after rIL-2 exposure, the LGL expressed only TCR gamma delta heterodimer in association with CD3 and alpha beta heterodimer expression could no longer be detected. The TCR/CD3 complex on these cells was demonstrated to be functional as anti-CD3 elicited an increase in cytoplasmic free calcium concentration, stimulated cytolytic activity, and stimulated granule enzyme secretion from the LGL. PMID- 2965724 TI - Characterization of a suppressor cell line which downgrades experimental autoimmune uveoretinitis in the rat. AB - A Ts lymphocyte line was isolated from spleens of rats primed with the retinal soluble Ag (SAg) in the anterior chamber of the eye. This line could inhibit in vitro SAg-driven proliferation of uveitogenic Th lymphocytes, in a radioresistant, Ag-independent manner. Adoptively transferred Ts line cells were found to downgrade experimental autoimmune uveoretinitis in actively immunized syngeneic recipients. The initial surface phenotype (OX8+) of the Ts line was unstable in vitro, however, the cells expressed suppressor function irrespective of phenotype. The mechanism of suppression did not appear to involve consumption of IL-2 or direct cytolysis of uveitogenic Th lymphocytes, but rather the production of a soluble suppressor factor. These findings may suggest an in vivo role for suppressor lymphocytes, capable of inhibiting primed Th cells, in the regulation of experimental autoimmune uveoretinitis. PMID- 2965723 TI - Bidirectionality of mixed lymphocyte stimulation (Mls) response. Effects of Mlsb stimulator cells on Mlsa helper cells. AB - The activation of BALB/c lymphocytes in the mixed lymphocyte reaction to Mls disparate APC has been shown to encompass up to 20% of the mature resting helper T lymphocyte population. In addition to these overtly Mls-responsive cells, our studies have revealed a second population that respond to the Mls difference of DBA/2 spleen cells in conjunction with the mitogen Con A. This part of the Mls response is therefore latent. As mitogen and Mls-stimulating effect act in synergy, it is likely that both stimuli act on the same cell, and hence the Mls effect can be regarded as a regulatory interaction between APC and Th cell. By use of congenic BALB.Mlsa mice, the regulatory effect has been mapped to the Mls locus. The regulatory influence has also been demonstrated in DBA/2 Th cells (Mlsa) stimulated simultaneously with mitogen and Mls-disparate (Mlsb) APC, consistently causing inhibition of mitogen-induced proliferation in this reverse Mls direction. This antagonistic effect has also been linked to the Mls locus. We conclude that the Mls reaction governed by the a and b alleles is bidirectional, producing synergy with class II-dependent activation signals in the direction of Mlsa----Mlsb, and antagonism in the direction Mlsb----Mlsa. Both the classical Mls and the reverse Mls effects have been demonstrated at the clonal level. These results are in accord with the previously proposed hypothesis that the Mls molecule serves as a down-regulatory stimulus in the activation of Th cells. Mls responses of Mlsb T cells are explained as the consequence of a diminished down regulation by Mlsa APC. Conversely, the reverse Mls response described here can be considered a consequence of inordinately high down-regulation of the Mlsa T cell responses by Mlsb APC. PMID- 2965725 TI - Lysis of cells infected with HIV-1 by human lymphocytes targeted with monoclonal antibody heteroconjugates. AB - The present study was undertaken to determine whether human PBL can be specifically focused to lyse cells infected with HIV-1 by mAb heteroconjugates that can bridge target and effector cells. A mAb directed against the central portion of HIV-1 glycoprotein gp110 was chemically cross-linked to a mAb directed against the CD3/TCR complex or to a mAb directed against the CD16 Fc gamma-R expressed on large granular lymphocytes (LGL). HIV-1-infected cells, but not uninfected cells, were found to be lysed to a greater extent by PBL in the presence of the gp110 X CD3 or the gp110 X CD16 antibody heteroconjugate than in the presence of the single antibodies or a mixture of the mAb comprising the heteroconjugates. Pretreatment of PBL with anti-CD3 or IL-2 augments their ability to lyse HIV-1-infected cells in the presence of the heteroconjugates. Lysis by anti-CD3-activated PBL in the presence of the gp110 X CD3 heteroconjugate was found to be mediated by CD8+-enriched T cells, whereas lysis by IL-2-treated PBL in the presence of the gp110 X CD16 heteroconjugate is mediated by PBL enriched for CD16+ cells, which are primarily LGL. Furthermore, PBL from asymptomatic, HIV-1-infected seropositive donors were found to be functional in lysing HIV-1-infected cells in the presence of the antibody heteroconjugates. Such antibody heteroconjugates, which can target T cells or LGL to lyse HIV-1-infected cells, may be of prophylactic or therapeutic value in HIV 1-infected individuals. PMID- 2965726 TI - Regulation of Fc receptor for IgE (CD23) and class II MHC antigen expression on Burkitt's lymphoma cell lines by human IL-4 and IFN-gamma. AB - The effect of rIL-4 on the expression of low affinity receptor for the Fc part of IgE (Fc epsilon R2/CD23) and class II MHC antigens on Burkitt's lymphoma (BL) cell lines was investigated. Some of the BL lines contained low percentages of CD23 and HLA-DQ-positive cells, but virtually all cells expressed HLA-DR. IL-4 induced CD23 and class II MHC Ag expression on 7 of 9 BL. Optimal CD23 and class II MHC expression was observed after 48-72 h of incubation. Induction of CD23 and class II MHC Ag in the BL cell line BL2 by IL-4 was confirmed at the specific mRNA level. Significant activation of HLA-DQ mRNA was obtained after 6 h of incubation with IL-4 and gradually increased during prolonged incubation. Maximal induction of mRNA transcription occurred after 48 to 72 h. Optimal induction of HLA-DR and CD23 transcription in BL2 was also observed after 48 to 72 h. The induction of CD23 and class II MHC Ag seems to be specific for IL-4, because rIL 1, rIL-2, rIFN-gamma, recombinant granulocyte-macrophage-CSF, and a commercial source of low m.w. B cell growth factor were ineffective. In addition, the expression of class I MHC Ag, the transferrin receptor, CD38, CD25, CD10, CD20, and CD21 were not affected by IL-4. Interestingly, IFN-gamma and PGE2 suppressed the IL-4-induced membrane expression of CD23 and class II MHC Ag in a dose dependent way. IFN-gamma also blocked IL-4-induced CD23 mRNA transcription in BL2 completely, whereas PGE2 (10(-7) M) was partially inhibitory. The induction of CD23 and class II MHC Ag by IL-4 required intact protein synthesis as shown by its inhibition by cycloheximide. These results indicate that the induction of CD23 and class II MHC Ag by IL-4 is regulated in a coordinated way. PMID- 2965727 TI - Immunosuppressive lymphokine derived from natural suppressor cells. AB - Cloned natural suppressor (NS) cells derived from spleens of total lymphoid irradiated BALB/c mice were incubated with the phorbol ester, PMA, and calcimycin for 4 h. After thorough washing, the induced NS cells were incubated in serum free medium for 24 h and the supernatants were collected. The supernatants suppressed the MLR between normal adult responder and stimulator spleen cells. There was no Ag specificity or H-2 haplotype restriction of the MLR suppression. The supernatants did not inhibit [3H]thymidine incorporation per se, because they did not suppress mitogen stimulation of spleen cells. Protease digestion of the supernatants removed the suppressive activity, and dialysis studies indicated that the molecular size of the suppressive factor was larger than 50,000 Da and smaller than 100,000 Da. The suppressive activity was stable at 56 degrees C, pH 2, for 1 h. Thus, NS cell clones can be induced to secrete an immunosuppressive lymphokine, NS factor. PMID- 2965728 TI - Comparative effects of tumor necrosis factor-alpha and IL-1 beta on mitogen induced T cell activation. AB - The effect of rTNF-alpha on human T cell function was examined and compared with that of rIL-1 beta by assessing the ability of each cytokine to support mitogen induced proliferation, IL-2 production, and IL-2R expression. TNF-alpha and IL-1 beta each enhanced DNA synthesis induced by PHA or immobilized mAb to the CD3 molecular complex. In addition, each cytokine increased the number of cells entering the G1 phase of the cell cycle and augmented IL-2R expression. The combination of optimal concentrations of these factors supported these responses to a greater extent than either cytokine alone, suggesting that T cell responsiveness is independently regulated by the action of at least two separate monocyte derived cytokines. Whereas TNF-alpha had little effect, IL-1 beta augmented IL-2 mRNA expression and IL-2 production by mitogen-stimulated cells. Furthermore, IL-1 beta enhanced proliferation with increasing length of culture. Whereas TNF-alpha also enhanced proliferation late in culture, it was less effective in this regard than IL-1 beta. Thus, IL-1 beta and TNF-alpha augment mitogen-induced T cell proliferation by increasing the number of cells initially activated and by promoting subsequent cell cycle progression. They differ, however, in their capacity to promote IL-2 mRNA and IL-2 production and therefore ongoing T cell proliferation. PMID- 2965729 TI - Trypanosoma cruzi-induced suppression of IL-2 production. I. Evidence for the presence of IL-2-producing cells. AB - Mice infected with the protozoan parasite Trypanosoma cruzi, the causative agent of human Chagas' disease, are profoundly immunodepressed in their response to various Ag and mitogens. A key factor in this immunosuppression is the essential inability to produce the T cell growth factor IL-2. In this study we demonstrate that this failure to produce IL-2 in response to mitogen stimulation is not the result of the absence of production of soluble or membrane-bound IL-1 by macrophages. Limiting dilution analysis of the precursor frequency of IL-2 producers suggests that an adequate number of precursors for IL-2 production are present in the spleens of infected mice, but that their activity may be regulated by suppressor cells. The presence of precursor cells for IL-2 production is supported by experiments showing that the combination of calcium ionophores and PMA elicits IL-2 production by spleen cells from both normal and T. cruzi infected mice. Although Con A can provide either of the signals necessary for IL 2 production, calcium flux or protein kinase C activation, to T cells from normal mice, Con A in combination with either calcium ionophore or phorbol ester failed to activate T cells from infected mice to produce IL-2. Preculture of spleen cells from infected mice for 48 to 72 h before addition of Con A results in near normal production of IL-2. This recovery of the capacity to produce IL-2 does not occur if parasite Ag is present during the preculture period. These results suggest that the inability of T cells from T. cruzi-infected mice to produce IL-2 in vitro in response to Con A is not due to the lack of IL-2-producing cells, but may be the result of the maturational state of the T cells or to the presence of a suppressor population. PMID- 2965730 TI - Trypanosoma cruzi-induced suppression of IL-2 production. II. Evidence for a role for suppressor cells. AB - Suppression of IL-2 production during experimental Chagas' disease accounts at least in part for the overall depressed state of the immune system in infected mice. The failure to produce IL-2 in response to mitogen stimulation is not the result of the lack of cells capable of producing IL-2, but appears to be due to regulation of IL-2 production by suppressor cells. This conclusion is supported by cell-mixing experiments where the ability of cells from infected mice to suppress normal spleen cell IL-2 production is evident. Although depletion of plastic and Sephadex G-10 adherent cells results in modest increases in IL-2 production by spleen cells from infected mice, even in the presence of normal adherent cells as a source of IL-1 producers, IL-2 production does not approach normal levels. Also, isolated macrophages are not by themselves suppressive for normal spleen cell IL-2 production, whereas plastic and G-10 nonadherent cells from infected mice are. Depletion of Thy-1+ and Ly-2+ cells not only completely abrogates the ability of spleen cells from infected mice to suppress normal IL-2 production, but results in a cell preparation which actually enhances IL-2 production. Anti-Ly-2 and C treatment of infected spleen cells also markedly enhances their ability to produce IL-2. These results indicate a major role for Ts cells in the regulation of IL-2 production, and a relatively minor role of macrophages as direct effector cells of suppression in this response. The ability to enhance IL-2 production in this system with PG synthesis inhibitors suggests a role for PG-producing cells such as macrophages in the suppressor mechanism, perhaps as inducers of the suppressor effector cells. PMID- 2965731 TI - Poly-N-acetyllactosamine structures on murine cell surface T200 glycoprotein participate in natural killer cell binding to YAC-1 targets. AB - Cell-surface murine T200 glycoprotein has been implicated in the binding of NK cells to certain susceptible tumor targets. The existence of poly-N acetyllactosamine structures on T200 glycoprotein and the ability of lactosamine type oligosaccharides to inhibit NK cell-mediated cytotoxicity suggest that these structures may also be important in NK-target binding. To further identify and characterize these structures, relevant saccharides and reconstituted membrane liposomes containing fractionated effector cell membrane proteins were tested for their ability to block conjugate formation. Under base line conditions, the majority of plastic-non-adherent, Percoll-fractionated, NK-enriched splenocytes that formed conjugates with NK-susceptible YAC-1 targets functioned as lytic effectors in a single-cell cytotoxicity assay. These effectors were blocked in their ability to bind to YAC-1 targets by the addition of N-acetyllactosamine [Gal(beta 1,4)-GlcNAc] and chitobiose [GlcNAc(beta 1,4)GlcNAc], but not by saccharides lacking lactosamine-type linkages. Liposomes prepared from octyl-beta D-glucopyranoside-extracted YAC-1 and NK-enriched effector cell membranes interfered with conjugate formation, whereas liposomes prepared from NK insensitive P815 cells were inconsequential. Surface radiolabeled effector cell membrane proteins were fractionated by tomato lectin-Sepharose 4B (poly-N acetyllactosamine-specific) column chromatography. Tomato lectin-bound material was enriched in a glycoprotein identical with T200, which, when incorporated into liposomes, was a potent inhibitor of effector-target binding. This inhibitory capacity was abrogated by treatment of liposomes with Ly-5 mAb (T200 mAb) or the lactosamine-specific enzyme endo-beta-galactosidase. When T200 was purified by mAb affinity chromatography and incorporated into liposomes, it was a potent inhibitor of conjugate formation, an effect that was blocked by pretreatment of T200-containing liposomes with Ly-5 mAb or endo-beta-galactosidase. These data provide additional evidence that T200 can mediate binding of NK cells to YAC-1 targets, and that poly-N-acetyllactosamine-type structures on NK cell surface T200 glycoprotein are important in the binding process. PMID- 2965732 TI - Defining research problems. PMID- 2965733 TI - The pediatric anesthesia team. PMID- 2965734 TI - Retinoblastoma: a case history. PMID- 2965735 TI - Evidence that activated mucosal T cells play a role in the pathogenesis of enteropathy in human small intestine. AB - T cells in explants of human fetal small intestine in organ culture were stimulated in situ with PWM or anti-CD3 antibody to test the hypothesis that activated T cells produce enteropathy in human small intestine. T cell activation was measured by the appearance of CD25+ cells in the lamina propria of the explants and IL-2 production into the organ culture supernatant. We have previously shown that the number of T cells in human fetal gut increased between 14 and 22 wk gestation. Accordingly, after the addition of PWM to cultured explants of fetal intestine the number of CD25+ cells in the lamina propria and the amounts of IL-2 secreted into the organ culture supernatant increased with the age of the explanted tissue. The addition of PWM also produced an age-related enteropathy, most noticeably crypt epithelial cell hyperplasia and villous atrophy, with relatively minor changes in 14-17-wk-old intestine but severe tissue damage in 18-22-wk-old fetal intestine. These enteropathic effects were also produced when mucosal T cells were activated with anti-CD3 mAb. Cyclosporin A completely inhibited the PWM-induced development of CD25+ cells and related tissue damage. These experiments show that activated T cells in human small intestine produce enteropathy. The model provides a new system with which to dissect the mechanisms of T cell-mediated intestinal damage. PMID- 2965736 TI - Heterogeneity of helper/inducer T lymphocytes. II. Effects of interleukin 4- and interleukin 2-producing T cell clones on resting B lymphocytes. AB - To compare the helper function of murine T cell clones that secrete IL-2 and IFN gamma (Th1 cells) or IL-4 and IL-5 (Th2), purified resting B cells were stimulated with F(ab')2 rabbit anti-mouse Ig (RAMG) and rabbit Ig-specific, class II MHC-restricted cloned T cells belonging to the two subsets. Both Th2 clones examined induced strong proliferative responses of B cells in the presence of RAMG, as well as the secretion of IgM and IgG1 antibodies. In contrast, the Th1 clones tested failed to stimulate B cell growth or antibody secretion. Th2 mediated B cell activation was dependent on IL-4 and IL-5, and was also inhibited by IFN-gamma or IFN-gamma produced by Th1 cells present in the same cultures. However, the failure of Th1 cells to help resting B cells could not be reversed with neutralizing anti-IFN-gamma antibody. In addition to this inhibitory effect, IFN-gamma was required for the secretion of IgG2a antibody, particularly when B cells were stimulated with polyclonal activators such as LPS. Finally, both sets of T cell clones secreted lymphokines when stimulated with purified B cells and RAMG. These experiments demonstrate that T cells that differ in lymphokine production also differ in their ability to help B cells as a result of cognate interactions at low concentrations of antigens. Moreover, IL-4, IL-5, and IFN gamma serve different roles in the T cell-dependent proliferative and differentiative responses of resting B lymphocytes. PMID- 2965737 TI - Human recombinant interleukin 4 induces Fc epsilon R2/CD23 on normal human monocytes. AB - rIL-4 (B cell stimulatory factor 1) induces the expression of Fc epsilon R2/CD23 on normal human monocytes (Mo). Fc epsilon R2/CD23 induction was detectable both by flow cytometry using anti-CD23 mAbs as well as soluble IgE, and by the immunoprecipitation with CD23-specific mAb or IgE of a 45-kD band from 125I lactoperoxidase-labeled Mo. Fc epsilon R2/CD23 was fully expressed after a 24-h incubation with rIL-4, and was still detectable after 72 h from the addition of IL-4. This effect was specific, because none of the other rILs tested (IL-1, IL 2, IL-3, IL-5, B cell stimulatory factor 2, granulocyte-macrophage colony stimulating factor, and IFN-gamma) could induce FC epsilon R2/CD23, either alone or in various combinations. No synergism was observed between IL-4 and other ILs. IFN-gamma was not able to inhibit the IL-4-induced expression of Fc epsilon R2/CD23 on Mo, neither when added to the culture together with IL-4, nor when added 36 h earlier. PMID- 2965739 TI - Thymus and autoimmunity. Transplantation of the thymus from cyclosporin A-treated mice causes organ-specific autoimmune disease in athymic nude mice. AB - Organ-specific autoimmune diseases such as gastritis, oophoritis, thyroiditis, or insulitis developed in athymic nu/nu mice after engraftment of the thymus from euthymic nu/+ mice treated with cyclosporin A (CsA), a potent immuno-suppressant. The development of autoimmune disease in the nu/nu mice was prevented by inoculation of thymocyte suspensions prepared from normal nu/+ mice, but not by thymocyte suspensions from CsA-treated nu/+ mice. Cotransplantation of normal nu/+ mouse thymus with CsA-treated thymus also suppressed the development of autoimmune disease. Inoculation of spleen cell suspensions prepared from normal adult nu/+ mice prevented autoimmune disease, but inoculation of those from newborn nu/+ mice did not. Thus, CsA appears to interfere selectively with the thymic production of certain suppressor T cells controlling self-reactive (autoimmune) T cells, allowing the latter to expand and cause autoimmune disease. PMID- 2965738 TI - T cell growth and differentiation induced by interleukin-HP1/IL-6, the murine hybridoma/plasmacytoma growth factor. AB - Interleukin-HP1 (HP1)/IL-6 is a 25-30-kD protein produced by macrophages, fibroblasts, and certain T cell lines. It was originally identified as a mouse growth factor for B cell hybridomas and plasmacytomas, and was recently shown to stimulate growth and differentiation of normal B cells. Here we demonstrate that, in the presence of lectins or anti-T cell receptor antibodies, HP1/IL-6 has a growth factor activity equivalent to that of IL-2 for mature thymic and peripheral T cells of both the L3T4+ and Lyt-2+ subsets. Contrary to IL-2 and IL 4, HP1/IL-6 was, however, not capable of supporting the growth of established T cell lines. In addition to its effects on T cell proliferation, HP1/IL-6 also enhanced the differentiation of mouse cytolytic T cell precursors in primary allogeneic mixed lymphocyte cultures. Fractionation of responding cell populations indicated that HP1/IL-6 was capable of restoring the response of accessory cell-depleted T cells to Con A. This observation suggests that the production of HP1/IL-6 by macrophages could, at least partly, explain their role in polyclonal T cell activation. PMID- 2965740 TI - Acquisition of an additional antigen specificity after mouse CD4 gene transfer into a T helper hybridoma. AB - We have transfected the mouse CD4 gene into a beef insulin (BI)-specific murine T helper hybridoma that lacks CD4 surface expression. The CD4-expressing transfectants have acquired an additional reactivity for pork insulin (PI), which was not detectable in the original recipient cell. The transfectants' response to PI can be completely abrogated by anti-CD4 antibodies. The transfected clone showed a 50-fold increased sensitivity towards BI in comparison to the same CD4- hybridoma. These experiments suggest that CD4 may be important in determining the antigen fine specificity and, therefore, may also play a role in altering the T cell repertoire. PMID- 2965741 TI - Antigen recognition by human T cell receptor gamma-positive lymphocytes. Specific lysis of allogeneic cells after activation in mixed lymphocyte culture. AB - These experiments were designed to define the ability of human TCR-gamma+ cells to recognize allogeneic cells. TCR-gamma+-enriched populations were obtained by treating peripheral blood E-rosetting cells with anti-CD4 and anti-CD8 mAbs. The resulting populations were CD2+4-8- expressed variable proportions of CD3+ cells (40-90%), and did not react with the WT31 mAb, which is specific for a framework determinant of the alpha/beta heterodimer that serves as receptor for antigen on most human T lymphocytes. After mixed lymphocyte culture with irradiated allogeneic cells for 7 d and 3 additional days in rIL-2 (100 U/ml), cells underwent proliferation in three of five individuals tested. In addition, MLC derived cells lysed 51Cr-labeled PHA-induced blasts derived from the allogeneic cells used as stimulator, but not allogeneic unrelated or autologous blast cells. No cytotoxicity against autologous or allogeneic target cells could be induced by culturing CD3+4-8-WT31- lymphocytes in MLC with irradiated autologous cells. Surface iodination of allogeneic MLC-activated CD3+4-8-WT31- cells followed by lysis in 1% digitonin and immunoprecipitation with anti-CD3 mAb indicated that the CD3-associated molecules consisted of a major 45-kD band and a minor band of 43 kD. Northern blot analysis showed that mRNA for the gamma chain was expressed at high levels, whereas mRNAs for alpha and beta chains were missing. These data support the notion that TCR-gamma rather than TCR-alpha/beta is expressed in allospecific CD3-4-8-WT31- cell populations. Clones were further derived from MLC stimulated CD3+4-8-WT31- populations. All the seven clones studied in detail maintained the surface phenotype as well as the cytolytic pattern of the original MLC populations, thus only specific allogeneic PHA-induced blasts were lysed. NK sensitive as well as NK-resistant tumor targets were variably susceptible to lysis; therefore, specific cytolytic activity against allogeneic cells was not necessarily linked to the expression of MHC-nonrestricted cytotoxicity against tumor cells. PMID- 2965743 TI - The ontogenesis of perception. PMID- 2965742 TI - What does infant perception tell us about theories of perception? AB - The article begins with a summary of eight well-substantiated conclusions drawn from research on perception in infants that have implications for theories of perception. The following section examines several traditional and modern theories of perception in the light of these conclusions. Finally, some "big issues" that have divided perceptual and cognitive theories are discussed, with the suggestion that research on infants can help reconcile the divided camps. Research on perception in young infants not only flourishes but also presents a challenge to all theories of perception. PMID- 2965744 TI - Perceptual completion of surfaces in infancy. AB - Two experiments provide evidence that 4-month-old infants perceive background surfaces as continuous behind occluding objects. Infants were shown a partly hidden background surface either for a brief period of familiarization (Experiment 1) or until they met a criterion of habituation (Experiment 2). The infants were then tested with nonoccluded surfaces that were either continuous or interrupted by a gap where the occluder had been. The infants in each study looked longer at the interrupted than at the continuous surface, relative to infants in baseline controls, which suggests that the partly hidden surface was perceived as continuous. Contrasting findings were obtained in a third experiment, in which infants were habituated to a partly hidden surface that stood in front of a background so that its edges were visible: Infants gave no evidence of perceiving the foreground surface as continuous behind the occluder. These experiments provide evidence that infants perceive a surface as continuous only if it serves as the background of a scene. The results are discussed in relation to figure-ground perception in pictures and surface layouts. PMID- 2965745 TI - Detection of the traversability of surfaces by crawling and walking infants. AB - In four studies we investigated the perception of the affordance for traversal of a supporting surface. The surface presented was either rigid or deformable, and this property was specified either optically, haptically, or both. In Experiment 1A, crawling and walking infants were presented with two surfaces in succession: a standard surface that both looked and felt rigid and a deforming surface that both looked and felt nonrigid. Latency to initiate locomotion, duration of visual and haptic exploration, and displacement activity were coded from videotapes. Compared with the standard, the deforming surface elicited longer latency, more exploratory behavior, and more displacement in walkers, but not in crawlers, suggesting that typical mode of locomotion influences perceived traversability. These findings were replicated in Experiment 1B, in which the infant was presented with a dual walkway, forcing a choice between the two surfaces. Experiments 2, 3A and B, and 4A and B investigated the use of optical and haptic information in detecting traversability of rigid and nonrigid surfaces. Patterns of exploration varied with the information presented and differed for crawlers and walkers in the case of a deformable surface, as an affordance theory would predict. PMID- 2965746 TI - Development of three-dimensional form perception. AB - In three experiments with infants and one with adults we explored the generality, limitations, and informational bases of early form perception. In the infant studies we used a habituation-of-looking-time procedure and the method of Kellman (1984), in which responses to three-dimensional (3-D) form were isolated by habituating 16-week-old subjects to a single object in two different axes of rotation in depth, and testing afterward for dishabituation to the same object and to a different object in a novel axis of rotation. In Experiment 1, continuous optical transformations given by moving 16-week-old observers around a stationary 3-D object specified 3-D form to infants. In Experiment 2 we found no evidence of 3-D form perception from multiple, stationary, binocular views of objects by 16- and 24-week-olds. Experiment 3A indicated that perspective transformations of the bounding contours of an object, apart from surface information, can specify form at 16 weeks. Experiment 3B provided a methodological check, showing that adult subjects could neither perceive 3-D forms from the static views of the objects in Experiment 3A nor match views of either object across different rotations by proximal stimulus similarities. The results identify continuous perspective transformations, given by object or observer movement, as the informational bases of early 3-D form perception. Detecting form in stationary views appears to be a later developmental acquisition. PMID- 2965747 TI - Contrast discrimination in human infants. AB - The ability to detect differences in spatial contrast is crucial to object recognition and identification. This ability is generally examined by measuring the contrast discrimination function. This function represents, for a variety of conditions, the smallest contrast difference required to discriminate otherwise identical patterns. We examined human infants' ability to discriminate patterns on the basis of differences in spatial contrast. The forced-choice preferential looking procedure was used to estimate contrast increment thresholds at a number of background contrasts. The Weber fractions of 6- and 12-week-old infants were about 1 log unit higher than adult values for background contrasts ranging from 0.14 to 0.55. Furthermore, the slopes of infants' discrimination functions were much shallower than those of adults. These age differences in contrast discrimination imply certain changes in the neural mechanisms that underlie contrast encoding. They also aid our understanding of the anomalies observed in early pattern vision. PMID- 2965748 TI - Detection of stimulus motion in 5-month-old infants. AB - Five-month-old infants were tested by the method of preferential looking for discrimination between a pattern undergoing oscillating apparent motion and an identical static pattern. Sensitivity to small spatial displacements was evident at temporal frequencies of 8 and 16 Hz. Preference for the moving display was related independently to the temporal frequency of oscillation and the magnitude of the spatial displacement. Preferences for the moving display increased asymptotically across spatial displacements from 11 to 89 arc min. Preferences peaked between temporal oscillation frequencies of 8 and 16 Hz. Preference was not related to the ratio of these two variables--velocity. The minimum displacement threshold of 7.36 arc min was found to depend on the size of the elements in the pattern and on the temporal frequency of oscillation. The results demonstrate that motion-sensitive mechanisms responsive to small spatial displacements are present at 5 months of age. PMID- 2965749 TI - Perception of biomechanical motions by infants: implementation of various processing constraints. AB - Geometry informs us that there exist a large number of possible connectivity patterns consistent with a point-light display of a person walking. Yet there is only one pattern consistent with a "stick figure" representation of the human form, and that pattern is uniquely specified by those pairwise connections that remain locally rigid. In this study, sensitivity to local rigidity in biomechanical displays was investigated in 3- and 5-month-old infants. The results of Experiment 1 revealed that by 5 months of age, infants discriminate a locally rigid point-light walker display from one in which local rigidity is perturbed. In Experiment 2 we tested infants' sensitivity to the same stimuli when those stimuli were inverted. Contrary to the preceding experiment, the results revealed no evidence of discrimination. Taken together, these findings suggest that infants are sensitive to local rigidity in biomechanical displays but that this sensitivity is orientation specific. Possible mechanisms for this specificity are discussed in the context of additional constraints on the processing of biomechanical displays. PMID- 2965750 TI - Object and observer motion in the perception of objects by infants. AB - Sixteen-week-old human infants distinguish optical displacements given by their own motion from displacements given by moving objects, and they use only the latter to perceive the unity of partly occluded objects. Optical changes produced by moving the observer around a stationary object produced attentional levels characteristic of stationary observers viewing stationary displays and much lower than those shown by stationary observers viewing moving displays. Real displacements of an object with no subject-relative displacement, produced by moving an object so as to maintain a constant relation to the moving observer, evoked attentional levels that were higher than with stationary displays and more characteristic of attention to moving displays, a finding suggesting detection of the real motion. Previously reported abilities of infants to perceive the unity of partly occluded objects from motion information were found to depend on real object motion rather than on optical displacements in general. The results suggest that object perception depends on registration of the motions of surfaces in the three-dimensional layout. PMID- 2965751 TI - Stimulus energy does not account for 2-month-olds' face preferences. AB - We examined the determinants of 2-month-olds' preferences among facelike and abstract patterns. Observed preferences were compared with the predictions of two preference models--one based on stimulus energy (as measured by the amplitude spectrum) and the other based on stimulus structure (as measured by the phase spectrum). It is known that the phase spectrum is the primary determinant of perceived identity to adults. Twenty-five 2-month-olds saw six pairings of four patterns: a schematic face, a lattice, a pattern composed of the amplitude spectrum of the lattice and the phase spectrum of the face, and a pattern composed of the amplitude spectrum of the face and the phase spectrum of the lattice. Only patterns with the face's phase spectrum look facelike to adults. Unlike the preferences of newborns (Kleiner, 1987), 2-month-olds' preferences could be predicted from the phase spectrum but not from the amplitude spectrum. In other words, the 2-month-olds preferred the patterns that looked facelike to adults. These results offer clear evidence that 2-month-olds' preferences for facelike patterns are not governed by stimulus energy. PMID- 2965752 TI - Infants' detection of visual-tactual discrepancies: asymmetries that indicate a directive role of visual information. AB - Infants' cross-modal functioning was investigated in two studies. In Study 1, 11 month-old infants were confronted with five different visual-tactual discrepancies created with a mirror arrangement. The infants' behavioral reactions to the discrepancies were compared with their behavior on matched control trials with a forced-choice judgement procedure. Infants detected discrepancies in which they saw an egg and felt a cube, saw a fur-covered cube and felt an egg, and saw a cross and felt a fur-covered cube. However, they provided no evidence that they detected discrepancies in which they saw a cube and felt a cross or saw a cube and felt a fur-covered cube. In Study 2, infants were confronted with discrepancies that were the converse of those which seemed to go unnoticed in Study 1: They saw either a cross or a fur-covered cube and felt a plain cube. Both of these new discrepancies were detected according to the forced-choice judgment procedure. The results indicate that texture as well as shape can serve as a basis for cross-modal matching for infants. The asymmetries in cross-modal matching that were observed across Studies 1 and 2 are interpreted as evidence that visual information plays a directive, goal-setting role for infants' manual explorations. PMID- 2965753 TI - The Ames window illusion: perception of illusory motion by human infants. AB - The sensitivity of human infants, 5 1/2-9 months of age, to the illusory oscillation of the Ames window was assessed in three experiments that employed some variant of the habituation-dishabituation and forced-choice preferential looking paradigms. In Experiment 1, three groups--5 1/2, 7 1/2, and 9 months of age--were given a visual choice between rotating rectangular and Ames windows after exposure to a rotating circular form. The two older groups preferred the Ames window. The results of Experiment 2 showed that this preference is not based on structural differences between the two windows. In Experiment 3, familiarization with an Ames window produced a preference for rotary motion while familiarization with a rectangular window produced a preference for oscillatory motion. These results suggest that sensitivity to the illusion emerges around 7 1/2 months of age, an outcome consistent with the emergence, at this time, of sensitivity to pictorial cues to depth. PMID- 2965754 TI - Dopamine D-1 receptor agonist stimulation of prolactin secretion in man. AB - SKF 38393, a selective D-1 dopamine receptor agonist, elevated plasma prolactin levels in eight patients with various neurological disorders. Growth hormone concentrations were unaffected by SKF 38393 administration. The results suggest that D-1 receptors may be involved in the regulation of prolactin secretion. PMID- 2965755 TI - Dose-ranging evaluation of the serotonin antagonist GR-C507/75 (GR38032F) when used as an antiemetic in patients receiving anticancer chemotherapy. AB - GR-C507/75 (GR38032F) antagonizes the 5-HT3 (serotonin) receptor and prevents cisplatin-induced emesis in animals. In this dose-ranging trial, 44 patients with cancer receiving chemotherapy known to produce nausea and vomiting (including cisplatin, cyclophosphamide, and doxorubicin) received three intravenous (IV) infusions of GR-C507/75 every two hours beginning 30 minutes before chemotherapy. Ten dosage levels were explored, ranging from 0.04 mg/kg to 0.35 mg/kg in each of the three infusions. Toxicities were mild and included sedation, dizziness, headache, transient elevations of SGOT or alanine aminotransferase (ALT), and dry mouth. No akathisia or acute dystonic reactions were observed. Antiemetic effects were seen in patients receiving cisplatin at 120 mg/m2. GR-C507/75 can be safely administered on this schedule at IV dosages up to 0.35 mg/kg in patients receiving chemotherapy. Further studies of this agent at higher dosages and by different schedules are appropriate. PMID- 2965756 TI - Three serotonin responses in cultured mouse hippocampal and striatal neurons. AB - Serotonin (5-HT) produced 3 different types of responses in neurons of mouse hippocampal and striatal cell cultures. These 3 responses have been characterized in terms of their pharmacological specificity, physiological mechanism, and dependence on cytoplasmic components. The most frequently observed response was inhibitory and was the result of a receptor-mediated activation of an inwardly rectifying potassium conductance. Typically, the response peaked within 1-3 sec of agonist application and did not exhibit desensitization. 5-Methoxy-N,N dimethyltryptamine also produced this response in both striatal and hippocampal cultures and had no effect on the other 5-HT currents observed in this study. The selective 5-HT agonists--8-hydroxy-2-(di-n-propylamino)-tetralin, 1-(m chlorophenyl) piperazine, and 1-(2-methoxyphenyl) piperazine--did not activate this outward current response. Methysergide did not block the 5-HT-activated outward current and often acted as an agonist. The response was lost in low series-resistance recordings which facilitate solution exchange between the patch electrode and the cell. The loss of this response was prevented by using high resistance patch electrodes, which retard this exchange. The 2 other responses described in this study were excitatory. They were seen less often than the inhibitory response. One of the excitatory responses was fast, with a time to peak of approximately 200 msec and a duration of 2-4 sec. The other was slow, with a time to peak of 7-10 sec and a duration of approximately 30-40 sec. Both of these responses were accompanied by a conductance increase. The fast excitatory response reversed at depolarized potentials and desensitized with a rate that varied with voltage. Metoclopramide and d-tubocurarine completely and reversibly blocked this fast excitatory response, while methysergide had no effect. The fast excitatory response was not lost during intracellular dialysis of cells in cultures from either striatum or hippocampus. In cultures from both brain regions, the slow excitatory response was blocked by methysergide. The slow excitatory response was lost even in patch-clamp recordings with high-resistance electrodes. This response was similar to responses to dopamine, norepinephrine, and forskolin, all of which are known to activate adenylate cyclase in the CNS. PMID- 2965757 TI - Acceptance of hepatitis B vaccine by medical and surgical residents. AB - To assess factors influencing acceptance of hepatitis B vaccine, 547 medical residents and 230 surgical residents were surveyed. The vaccination rate among 315 (58%) medical residents who responded was 46%; for 124 (54%) surgical residents who responded it was 76%. Most medical (93%) and surgical (94%) residents who were vaccinated believed they were at risk of hepatitis B virus infection. Among unvaccinated medical residents, 71% indicated concern about vaccine-related side effects, including potential but unknown reactions (58%) and possible transmission of AIDS (37%) and hepatitis (16%). Unvaccinated surgical residents were also concerned about side effects (64%). Stepwise discriminant function analysis revealed that medical residents were vaccinated if they were concerned about risk of exposure to hepatitis B virus and the chronic complications of infection and if they had received hepatitis B immune globulin and influenza vaccine. Surgical residents were vaccinated if they believed hepatitis B vaccine was efficacious, but were not vaccinated if they believed hepatitis B virus infection was not serious. PMID- 2965758 TI - Back pain and epidural spinal cord compression. AB - The physician must have a high index of suspicion to detect SCC early in patients with malignancy. Back pain is the first symptom in almost all patients, and the diagnosis should be considered for all older patients with back pain. Asking about back pain should be a routine part of the review of systems, especially for patients with known malignancies. Clinically, it is impossible to tell whether or not a patient who has back pain and cancer has epidural SCC. Patients may be stratified as to the likelihood of SCC using the history and physical examination, but the diagnosis relies on radiographic visualization of the spinal cord. It may be acceptable to closely follow patients with normal neurologic examinations and normal plain films, but even this is controversial and includes only a minority of patients. Myelography remains the test of choice. MRI will play an increasingly important role in the future, but has not yet been systematically evaluated. The best therapeutic approach is not clear, but standard treatment is only about 50% effective in all cases. At present, radiation therapy is the treatment of choice for many patients, in particular those who are ambulatory at diagnosis. Anterior resection with vertebral body reconstruction is an exciting approach and may substantially improve the prognosis for patients who are paraparetic or paraplegic. It is important to attempt to choose for each patient the diagnostic and therapeutic options offering the best chance for comfort and preservation of function. The decision of how or even whether to treat is multifactorial and is more complicated than the determination of simply whether or not compression is present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965759 TI - Keratan sulfate is a component of proteoglycans in the compressed region of adult bovine flexor tendon. AB - A monoclonal antibody (ET-4-A-4) was used to identify keratan sulfate (KS) as a constituent of bovine flexor tendon. KS was present in at least 500-fold higher amounts in the fibrocartilaginous region of the tendon that is subjected to compressive forces in vivo (mean = 0.03% of tissue dry weight) than in the more proximal regions subjected only to tensional forces. The KS was associated with proteoglycans of three size categories that could be separated by Sepharose CL-4B chromatography. The largest proteoglycan (Vo) contained approximately 4% KS and was predominant in the surface region of the tendon subjected directly to compressive forces. A population of somewhat smaller molecules (Kav approximately equal to 0.3) contained less than or equal to 20% KS and proportionately less chondroitin sulfate (CS) or dermatan sulfate (DS). The KS chains of this population were not digested by keratanase. This population was dominant in tissue from the middle layer of the fibrocartilaginous region. A third population of small KS proteoglycans or fragments (Kav approximately equal to 0.6) comprised 40% of the KS found in the deepest layer of the compressed tendon region. This smaller component was independent of the small DS proteoglycans. PMID- 2965760 TI - B-lymphocyte responses in the large intestine and mesenteric lymph nodes of mice infected with Eimeria falciformis (Apicomplexa). AB - B-cell responses of 3 immunoglobulin isotypes (IgA, IgG, and IgM) were investigated in the large intestine and mesenteric lymph nodes (MLN) of naive or immune mice after inoculation of oocysts of Eimeria falciformis. Primary and anamnestic IgA and IgG lymphocyte responses to E. falciformis occurred in the large intestine of nonimmune and immune mice, respectively. IgA-containing lymphocytes (IgAc) were the largest population of responding B cells in the large intestine. In infected mice, IgAc accumulated in the apical portion of the lamina propria, whereas IgG-containing lymphocytes (IgGc) were more numerous at the base of the lamina propria. No significant increase in the number of IgM-containing lymphocytes (IgMc) was observed in the lamina propria of the large intestine. Primary but no anamnestic B-cell responses occurred in the MLN, and immune mice actually had reduced numbers of IgAc and IgGc in the MLN when compared with naive mice. IgGc were the largest population of responding B cells in the MLN. Thus, IgAc appear to accumulate preferentially at the site of parasite development, whereas IgGc are primarily localized deeper in the lamina propria of the large intestine and in the draining lymph nodes of mice infected with E. falciformis. PMID- 2965762 TI - Papuloerythroderma in a woman. PMID- 2965761 TI - Effects of age on contractile and enzyme-histochemical properties of fast- and slow-twitch single motor units in the rat. AB - 1. Contractile, enzyme-histochemical and morphometrical properties of muscle fibres were studied in single motor units of tibialis anterior (t.a.) and soleus muscles in young (3-6 months) and old (20-24 months) male albino rats. The technique of measuring glycogen depletion as a marker of previous muscle contraction was used for direct correlation of enzyme-histochemical and contractile parameters within single motor units of the fast- and slow-twitch type. 2. In t.a., the fast-twitch motor units covered 18 +/- 9 and 22 +/- 16% (P = not significant, n.s.) of t.a. cross-sections, included 148 +/- 65 and 162 +/- 63 muscle fibres per unit (P = n.s.) and had a cross-sectional area of 0.50 +/- 0.32 and 0.44 +/- 0.22 mm2 (P = n.s.) in the young and old animals, respectively (means +/- S.D.). 3. In soleus, the slow-twitch motor units covered 53 +/- 11 and 71 +/- 12% (P = n.s.), included 55 +/- 10 and 83 +/- 13 muscle fibres per unit (P less than 0.01) and had a total cross-sectional area of 0.14 +/- 0.02 and 0.22 +/ 0.06 mm2 (P less than 0.01) in the young and the old animals, respectively. The calculated number of motor units in soleus accordingly decreased (P less than 0.01) from 49 +/- 10 in the young to 29 +/- 10 in the old animals resulting in a loss of muscle fibres and an increased innervation ratio in old age (mean +/- S.D.). 4. Clusters of more than three muscle fibres were rarely seen in any of the glycogen-depleted motor units in either the young or the old animals. However, in the slow-twitch motor units of old animals the muscle fibres were less randomly distributed within the motor unit territory (P less than 0.05), indicating a denervation-reinnervation process. 5. The contraction and half relaxation times of the isometric twitch were significantly prolonged in old age. In 274 randomly isolated single motor units of t.a. the contraction time increased from 13 +/- 1 in young animals to 17 +/- 3 ms in old ones and the half relaxation time from 12 +/- 2 to 16 +/- 5 ms (P less than 0.01 in both cases). In 236 randomly isolated soleus single motor units, the contraction and half relaxation times increased (P less than 0.001) from 24 +/- 5 to 31 +/- 7 ms and from 26 +/- 8 to 35 +/- 9 ms, respectively (mean +/- S.D.).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2965763 TI - In vitro characterization of monoaspartyl chlorin e6 and diaspartyl chlorin e6 for photodynamic therapy. AB - The characteristics of two new chlorin photosensitizers were studied in cell culture by determining phototoxicity, subcellular localization, and photophysical properties. Monoaspartyl chlorin e6 (MACE) and diaspartyl chlorin e6 (DACE) are new photosensitizers that show promise for use in photodynamic therapy. These chlorins are pure, monomeric compounds as determined by high-pressure liquid chromatography. Both compounds absorb substantially at a longer wavelength (664 nm) than does dihematoporphyrin ether-ester (DHE). Tumor diagnosis with the use of fluorescence should be facilitated due to the purity of the compounds and the single fluorescence emission peak. Phototoxicity dose-response curves of the sensitizers were completed using a standard clonogenic assay to determine cell viability. The chlorins showed good sensitizing capabilities with light. In addition, subcellular localization of MACE, DACE, and DHE was studied using fluorescence microscopy. Whereas DHE was located throughout the cytoplasm, the primary site of localization of the chlorins appeared to be in the lysosome. The results demonstrate that MACE and DACE are effective photosensitizing agents in vitro and compare favorably to DHE. PMID- 2965764 TI - EIa-mediated stimulation of the adenovirus EIII promoter involves an enhancer element within the nearby EIIa promoter. AB - The transcriptional induction of adenovirus early genes by the viral immediate early gene EIa constitutes an attractive model system for the study of control mechanisms involved in eucaryotic promoter function. The EIa-mediated activation of the divergently transcribed EIIa early (EIIaE) and EIII promoters was investigated in experiments in which recombinant plasmids containing the entire EIIa-EIII control region were cotransfected with a plasmid expressing the EIa 13S mRNA. First, both promoters were activated by low levels of EIa, but the extent of EIII induction decreased with increasing EIa concentrations, whereas EIIaE stimulation remained unchanged. Second, transcriptional analysis of deletion mutants revealed that an element of the EIIaE promoter contributed to maximal EIa responsiveness of the nearby EIII promoter. This element, located between positions -82 and -71 with respect to the EIIaE major cap site, corresponded to the central portion of an EIa-dependent enhancer, originally mapped between about -110 and -50 (P. Jalinot and C. Kedinger, Nucleic Acids Res. 14:2651-2669, 1986). The implication of these observations in the coordinate expression from the EIIaE and EIII promoters during lytic infection is discussed. PMID- 2965765 TI - Simian retrovirus D serogroup 1 has a broad cellular tropism for lymphoid and nonlymphoid cells. AB - Simian acquired immunodeficiency syndrome is a fatal immunosuppressive disease caused by type D retroviruses such as simian acquired immunodeficiency syndrome retrovirus type 1 (SRV-1). The disease is characterized by generalized lymphadenopathy, opportunistic infections, and lymphoid depletion with defects in both humoral and cell-mediated immunity. To understand how SRV-1 infection relates to the immune defect, we studied in vivo-infected lymphocytes from SRV-1 positive macaques with and without clinical signs of immunosuppressive disease. B and T helper/inducer and T suppressor/cytotoxic lymphocytes were purified by panning or by flow cytometry. Neutrophils were purified by dextran sedimentation, and platelets were purified by low-speed centrifugation. In vitro infection studies were also done with HUT78, H9, K562, rhesus lung fibroblast, rhesus monkey kidney, and bat lung cells. SRV-1 in lymphocytes or culture supernatants was detected by the induction of syncytia in cocultivated Raji cells and was confirmed by immunofluorescence, electron microscopy, or reverse transcriptase assay. We found that B and T helper/inducer lymphocytes were infected in all animals tested. The number of infected T suppressor/cytotoxic cells was generally lower than that of the other cell subsets, and not all animals in this subset had SRV-1 infections. All other cells exposed in vitro to SRV-1, except bat lung cells, were able to be infected. These findings show that SRV-1 has a broad cell tropism for lymphoid and nonlymphoid cell types. PMID- 2965767 TI - [Effects of sevoflurane anesthesia and surgery on plasma beta-endorphin-like immunoreactivity and prolactin levels]. PMID- 2965766 TI - Prediction of similar transforming regions in simian virus 40 large T, adenovirus E1A, and myc oncoproteins. AB - Regions containing similar elements of primary and predicted secondary structure were identified in simian virus 40 large T, adenovirus E1A, c-myc and v-myc proteins by a computer program with a set of highly specific, complex pattern descriptors. In all cases these regions were localized in domains of the respective proteins known to be required for transforming activity. We suggest that these apparently structurally similar regions may mediate a common biological function. PMID- 2965768 TI - [Two-color flow cytometry analyses of peripheral blood lymphocytes (PBL) and lymphokine-activated PBL in gastric cancer patients]. AB - A two-color flow cytometric analyses of the PBL of 51 patients (24 resectable, and 27 nonresectable), and of 30 healthy controls has been completed. The proportion of antigens, on the PBLs, defined by combinations of anti-Leu 2a and anti-Leu 15, anti-Leu 3a and anti-Leu 8, anti-Leu 3a and anti-HLA-DR, anti-HLA-DR and anti-Leu 2a and, anti-HLA-DR and anti-IL 2R, was not significantly different among the three groups. TCGF increased the proportion of Leu 2a+ Leu 15-, Leu 3a+ Leu 8-, HLA-DR+ Leu 2a+, Leu 3a+ HLA-DR+ and, HLA-DR+ IL 2R+ cells and reduced the proportion of Leu 2a+ Leu 15+ cells in gastric cancer patients, while the cultivation by IL 2 (2 weeks) increased the proportion of Leu 3a+ Leu 8-, HLA-DR+ IL 2R+ and Leu 3a+ HLA-DR+ cells, and did not change the proportion of Leu 2a+ Leu 15- and Leu 3a+ Leu 8+ cells, and decreased the proportion of Leu 2a+ Leu 15+ cells. These results, taken together with our previously published studies, suggest that the TCGF-activated Leu 2a+ Leu 15- subset but not the Leu 2a+ Leu 15+ subset serves as a phenotypic marker of suppressor-effector T cells, and that the IL 2-activated Leu 3a+ Leu 8- subset contained killer T cells against tumor cells in our experimental systems. PMID- 2965769 TI - [Nurse manager of home for the elderly--living together happily at home for the disabled elderly]. PMID- 2965770 TI - [Hemodynamic effect of a combination of prazosin with cardiac depressants in the treatment of hypertension]. AB - Arterial blood pressure and central hemodynamic changes were assessed in 44 patients with essential hypertension, uncontrollable by prazosin alone, after propranolol (anaprilin), celiprolol, clonidine (clofelin) or verapamil were added to the treatment schedule. A selective beta 1-adrenoblocker, celiprolol, is shown to be the best option in prazosin-treated patients with reflex increment of heart rate and stroke volume. The prazosin-verapamil combination is also justified in cases of moderately increased pulse rate, while propranolol and clonidine produce no additional hypotensive effect when combined with prazosin. PMID- 2965771 TI - [Results of a 2-year examination of patients with essential hypertension and left ventricular hypertrophy]. AB - A study of 52 middle-aged males with essential hypertension, unaccompanied by cardiovascular disorders, is reported. The patients were divided into 2 groups with respect to the type of treatment they received: 30 patients on beta adrenoblockers and 22 patients on methyldopa. The patients were treated continuously for 2 years under one-dimensional echocardiographic control. The greatest drop in arterial blood pressure and ventricular septum thickness was seen within the first 6 months of treatment, while posterior-wall hypertrophy disappeared altogether. Although there was no further drop in blood pressure after 6 months, the thickness of the ventricular septum remained reduced for at least 2 years. Regression of left-ventricular hypertrophy was not accompanied with any deterioration of left-ventricular performance, nor was it related to additional use of vasodilators. PMID- 2965772 TI - [Diastolic relaxation and left ventricular blood filling in patients with hypertension]. AB - Disorders of left-ventricular diastolic relaxation and blood filling in patients with first- and second-stage essential hypertension are shown, by means of echo-, radio- and apexcardiography, to be due to the effects of arterial blood pressure, hemodynamic changes, heart rate and the magnitude of left-ventricular hypertrophy. Antihypertensive treatment, affecting major parameters of disease, brings about the recovery of the heart's diastolic function. PMID- 2965773 TI - [Concentration of the atrial natriuretic factor in the atria and blood plasma in the development of hypertension in spontaneously hypertensive rats]. AB - The contribution of atrial natriuretic factor to the development of spontaneous arterial hypertension was investigated in rats (the SHR line). Prehypertensive atrial and plasma levels of a natriuretic factor were similar in SHR to those of the control WKY rats. In fully-developed hypertension, the factor content decreases in left-atrial wall and increases in the plasma. It is assumed that elevated arterial blood pressure affects myo-endocrine left-atrial cells, leading to hypersecretion of the natriuretic factor. This kind of secretory response might be an adaptive/compensatory mechanism of blood pressure control. PMID- 2965774 TI - Isolation and characterization of amyloid P component from Alzheimer's disease and other types of cerebral amyloidosis. AB - The presence of amyloid P-component (AP) within cerebral amyloid deposits was investigated by means of biochemical and immunocytochemical methods. Immunoperoxidase on formalin-fixed, paraffin-embedded tissue sections from Alzheimer's Disease, Down's Syndrome, asymptomatic age-related cerebral amyloidosis, sporadic cerebral amyloid angiopathy, hereditary cerebral hemorrhage with amyloidosis-Icelandic type, and hereditary cerebral hemorrhage with amyloidosis-Dutch type revealed the presence of AP in the affected vessel walls in all cases, and in parenchymal deposits resembling neuritic plaques of Alzheimer's disease, sporadic cerebral amyloid angiopathy, and hereditary cerebral hemorrhage with amyloidosis-Dutch type. A short digestion of tissue sections with pepsin was required for immunodetection of AP in these latter structures. After extraction of leptomeningeal amyloid fibrils, AP was characterized by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, Western blot, gel chromatography, and partial amino acid sequencing. Our results indicate that: (a) AP from cerebral amyloidosis has similar biochemical properties and homologous amino terminal sequence to AP from systemic amyloidosis; (b) AP is associated to a variety of brain amyloid deposits regardless of their chemical nature. The presence of AP, a serum protein, within the brain parenchyma points to an impairment of the blood-brain barrier in these diseases. PMID- 2965775 TI - Effects of estrogen and growth hormone on steroid sulfatase activity and estrogen binding in rat liver. AB - It is now well established that the activity of certain liver enzymes displays sex differences and that administration of human growth hormone to male rats alters the liver metabolism in a "female" direction. In this work we studied steroid sulfatase activity and binding of estradiol-17 beta in livers from intact rats and found a sex difference, with considerably higher enzyme activity in male as compared to female liver tissue. Continuous infusion of native and recombinant human growth hormone and estradiol-17 beta to male rats reduced sulfatase activity to "female" levels. A specific binding of estradiol-17 beta with receptor properties was found in the rat livers, but the concentration of binding sites did not change after administration of growth hormone or estradiol in this group of intact animals. Our data confirm previous reports that continuous administration of human growth hormone "feminize" liver metabolism, and since estradiol was found to have an identical effect on sulfatase activity it is suggested that the effect of estradiol-17 beta in this respect may be indirect, mediated via an altered secretory pattern of rat growth hormone. PMID- 2965776 TI - Action of interferon-alpha on hairy cell leukemia: expression of specific receptors and (2'-5')oligo (A) synthetase in tumor cells from sensitive and resistant patients. AB - IFN-alpha induces tumor regression with a high percentage of complete remissions in hairy cell leukemia. We have recently reported that hairy cells express specific IFN-alpha receptors which are down-regulated upon therapy. We show here that the activity of 2-5 A synthetase, an IFN-induced enzyme, is 2 to 7-fold stimulated in hairy cells from responsive patients within 12-24 h after the first IFN dose. This in-vivo enzyme induction paralleled the kinetics of receptor down regulation. In one patient who was unresponsive to IFN-alpha treatment neither expression of IFN-alpha receptor nor change in 2-5 A synthetase were expressed and modulated as in sensitive patients. In-vitro treatment of hairy cells with recombinant interferons showed that IFN-beta 1 was able to induce this enzyme to the same extent than IFN-alpha 2, whereas IFN-gamma was inactive despite the presence of specific IFN-gamma receptors. Our results indicate that IFN-alpha can exert its therapeutic effects by acting directly on the hairy cells through interaction with surface membrane receptors. Induction of 2-5 A synthetase can be one of the steps involved in this action. However, both mechanisms, as well as receptor down-regulation are not sufficient to explain the responsiveness of hairy cell leukemia to IFN-alpha. PMID- 2965777 TI - Preferential in-vitro growth and expansion of leukemic T lymphoblasts. AB - An in-vitro culture system was used to selectively grow malignant cells from the bone marrow of a patient with acute T-lymphoblastic leukemia. Molecular analysis of DNA extracted from the bone marrow cells before culture showed the presence of both rearranged and germ line patterns for the T-cell beta receptor (CTB) gene, and chromosomal analysis revealed the presence of a major and a minor abnormal clone. The cells were cultured in RPMI medium supplemented with 20% fetal calf serum, 2% lymphocyte conditioned medium, L-glutamine and antibiotics. The presence of malignant cells in the cultured population was confirmed by morphologic, molecular probing and cytogenetic analysis. After four weeks in culture, DNA extracted from the cultured cells showed only the rearranged pattern for the CTB gene. Chromosomal analysis of the same cultured sample revealed only the presence of the initially predominant abnormal clone. Shortly thereafter, analysis of fresh uncultured bone marrow cells from the patient in relapse revealed that the same chromosomally abnormal clone also predominated in vivo. Thus, our results demonstrate the selective nature of this culture system and its ability to amplify leukemic T-lymphoblasts. This culture system is also useful for detecting occult malignant cells in histologically normal bone marrow. PMID- 2965778 TI - Mitogenic stimulation of malignant B cells CLL: diminished in-vitro stimulation with anti-CR1 antibodies. AB - Peripheral B lymphocytes of five CLL patients were tested in a radioimmunoassay to determine the density of the C3b receptor (CR1) and the cells were assayed for their ability to mature into IgM secreting cells after in-vitro culture with a combination of Pokeweed Mitogen (PWM) and antibodies directed against CR1. Despite the presence of normal amounts of CR1 on the leukemic B cells, crosslinking of these receptors by anti-CR1 antibodies stimulated only a fraction of the leukemic cell population to differentiate into IgM secreting cells. These results add to the partial functional impairment of CLL-B cells. PMID- 2965779 TI - Oligosaccharide components of surface glycoproteins from leukemic lymphocytes of chronic lymphocytic leukemia and non-Hodgkin's lymphoma. AB - Oligosaccharide components of surface glycoproteins of leukemic cells have been studied in a group of eight patients with chronic lymphocytic leukemia (CLL) and non-Hodgkin's lymphoma (NHL). Labelled surface glycoproteins were separated by electrophoresis and treated with mild alkaline borohydride, prior to exclusion chromatography on Sephadex G-50. Five peaks were detected, of which peaks I and II consisted largely of N-linked chains and peaks III and IV of O-linked chains. Peak V contained monosaccharides and non-carbohydrate material. On both peripheral blood and lymph node cells of the CLL and nodular poorly differentiated lymphocytic lymphoma (NPDLL) patients studied, glycosylation was altered on leukocyte-common antigen (L-CA) forms of different relative molecular mass (Mr). The ratio of peak II oligosaccharides to those of peaks III and IV in different forms of L-CA correlated closely with Mr of the L-CA. Glycosylation of L-CA on cells of two patients with diffuse large cell lymphoma differed from that of the CLL and NPDLL patients, but was also to some extent related to Mr. Most of the oligosaccharides on the large sialoglycoprotein were O-linked, only a minor amount of N-linked being detected. Two surface glycoproteins of Mr 87 and 83 k differed markedly in their content of O- and N-linked chains. The specific glycosylation patterns described may have a role in control of cell behaviour and disease patterns of leukemia and lymphoma. PMID- 2965780 TI - Applying vowel sounds to command support systems for motor-impaired people. PMID- 2965781 TI - [Regional problems of the health of the population]. PMID- 2965783 TI - DNA damage and mutagenesis of lambda phage induced by gamma-rays. AB - Lambda phage DNA was gamma-irradiated in aqueous solution and the amount of radiation-induced strand breakage [double- and single-strand breaks and alkali labile sites (dsb, ssb, als)] was determined. Twice as much minor structural damage (ssb and als) per lethal hit was found in this DNA compared with DNA from irradiated phage suspensions. The in vitro irradiated DNA was re-packaged into infectious particles ('pack-phage'). The induction of mutations in the cI or cII cistron was scored using SOS-induced host cells. The in vitro prepared particles were found to have second-order kinetics for mutagenesis induced by gamma-rays indicating that two pre-mutational events were necessary to produce a mutation. In contrast, bacteria-free phage suspensions ('lys-phage') showed single hit kinetics for mutagenesis after irradiation. The increase in the mutation rate in the phage particles was mainly due to minor lesions, i.e. ssb, als and unidentified base damage. In lys-phage, mutagenesis might be enhanced by clustered DNA damage--a configuration which does not exist in pack-phage. The loss of infectivity was analysed in comparison with the extent of structural damage. All lesions investigated here contributed to biological inactivation. Minor lesions were tolerated by lambda phage to a limited extent. Major lesions, such as dsb, contributed most to the loss of infectivity and can be considered as lethal events. PMID- 2965782 TI - Evolution of the primary and secondary structures of the E1a mRNAs of the adenovirus. AB - In this paper we investigate and compare (evolutionary) patterns in the primary and secondary structure of four homologous E1a mRNAs of the adenovirus. Our main results are as follows: (1) The similarity of the coding regions of the mRNA sequences reflects both similarity in function (i.e., oncogenicity) and evolutionary divergence. (2) The similarity of the leader and the trailer regions reflects host specificity (i.e., human or simian) and must therefore arise from convergence. (3) Minimal energy foldings of the mRNAs show similar secondary structures (in particular around the splice sites). The conservation of pre-mRNA secondary structure shows that mRNAs are subject to selection constraints in addition to those associated with proteins. (4) The conserved secondary (helical) structures consist of nonhomologous subsequences, i.e., shifts have occurred. The observed shifts near the splice sites seem to be the simplest way of dealing with the dual constraints. PMID- 2965784 TI - Biochemical changes in rhesus monkey during the first days after streptozotocin administration are indicative of selective beta cell destruction. AB - Hormonal and glycemic changes in 22 rhesus monkeys were characterized during the first days after treatment with streptozotocin (STZ) (45 to 55 mg/kg, administered intravenously [IV]). Almost half (10/22) of the monkeys developed insulin-dependent diabetes mellitus (STZ-IDDM) within five days following injection. Four of the remaining monkeys did not become insulin dependent for at least 6 months after STZ treatment, during which time they were considered non insulin-dependent, and eight monkeys never required exogenous insulin. In the STZ IDDM group, plasma immunoreactive c-peptide (IRC-P) levels fell by three hours after STZ from a mean +/- SEM of 252 +/- 82 to 101 +/- 45 pg/mL, as glucose and immunoreactive glucagon (IRG) levels increased from 65 +/- 3 and 120 +/- 37, respectively, to 336 +/- 43 mg/dL and 234 +/- 52 pg/mL, respectively. Between six and 30 hours after treatment, IRC-P increased to a peak of 1,561 +/- 360 pg/mL before falling permanently to less than 60 pg/mL by 66 hours. During this period, glucose and IRG responded in a reciprocal fashion by falling and then increasing to levels above 300 mg/dL and 300 pg/mL, respectively, by 66 hours. In the non insulin-dependent diabetes mellitus (STZ-NIDDM) group, no clear reciprocal relationship between IRC-P and glucose and IRG was obtained. In nine additional monkeys subjected to total pancreatectomy (Px), IRC-P and IRG levels fell immediately and permanently by greater than 90% and 75%, respectively. Levels of immunoreactive somatostatin increased steadily over the initial 96 hours following STZ, but did so both STZ-IDDM and Px monkey groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965785 TI - [Chromium and nickel salts: a cause of allergic contact dermatitis from detergents]. PMID- 2965786 TI - [Three cases of thoracic or thoracoabdominal aortic aneurysm successfully treated with cell saver]. AB - During a three-month period from January to March 1986, reconstructive surgery was performed on three patients with thoracic or thoracoabdominal aortic aneurysm using a temporary bypass procedure. To reduce the amount of blood transfusion required during operation, an autologous blood recovery system, Cell Saver, was used in these three cases. The amount of blood loss, including the blood passing through the Cell Saver, was 7,020g in Case 1, 6,600g in Case 2 and 16,700g in Case 3. The amount of blood transfusion given to these three cases was 1,400g, 4,400g and 6,800g, respectively. The results indicated that the ratio of transfused blood amount to blood loss during operation was successfully reduced to a level of 20-67% of two other cases with thoracoabdominal aortic aneurysm, operated upon without using Cell Saver. Cell Saver was especially effective in Case 3 in which the blood for transfusion was restricted, for it was difficult to collect enough donors to complete operation because of rare blood type of Rh(-) in this case. That is, Cell Saver withdrew 15,000ml of blood through the sucker from the patient and returned it as concentrated and washed RBC during operation. PMID- 2965787 TI - A novel high affinity class of Ca2+ channel blockers. AB - Benzolactams (HOE 166 and analogs) form a new class of molecules acting on the 1,4-dihydropyridine-sensitive L-type Ca2+ channels. The main binding properties of HOE 166 and analogs to rabbit skeletal muscle membranes are as follows. (i) The compounds have a specific binding site to which they associate with a high affinity (0.25 nM for HOE 166). (ii) Unlabeled HOE 166 and analogs completely inhibit 1,4-dihydropyridine binding [(+)-[3H]PN 200-110] in a competitive way. (iii) Affinity values measured for HOE 166 inhibition of (+)-[3H]PN 200-110 (K0.5 = 0.25 nM and K1 = 0.55 nM) and of [3H]HOE 166 binding (K0.5 = 0.5 nM) are in good agreement. They also fit with results from direct binding experiments with tritiated HOE 166 (Kd = 0.27 nM) and from kinetic experiments (Kd = 0.39 nM). (iv) HOE 166 completely inhibits the specific binding of other classes of Ca2+ channel antagonists such as phenylalkylamines [(-)[3H] desmethoxyverapamil], benzothiazepines (d-cis-[3H]diltiazem), diphenylbutylpiperidines ([3H]fluspirilene), and [3H]bepridil. In all these cases the binding inhibition is of a noncompetitive nature. (v) The maximum binding capacity for [3H]HOE 166 binding to transverse tubule membranes, 65 pmol/mg of protein, is the same as that found for other classes of Ca2+ channel antagonists. 45Ca2+ uptake experiments performed with the rat aortic cell line A7r5 and the insulin secreting cell line RINm5F demonstrate that HOE 166 and analogs fully inhibit the 1,4-dihydropyridine-sensitive 45Ca2+ influx elicited by depolarization. There is a good correlation between inhibitory potencies of compounds in the HOE 166 series measured on (+)-[3H]PN 200-110 binding to A7r5 membranes and on the activity of Ca2+ channels followed by 45Ca2+ fluxes with the same cells. Structure-function relationships of HOE 166 and analogs for Ca2+ channel blockade in A7r5 and RINm5F cells were also in good correlation. Finally, voltage-clamp experiments confirmed that voltage-dependent L-type Ca2+ channels are completely blocked by 100 nM HOE 166 even at a membrane potential held at -80 mV. PMID- 2965788 TI - Role of epoxide in neocarzinostatin chromophore stability and action. AB - To determine the role of the epoxide moiety of the nonprotein chromophore of the antitumor antibiotic neocarzinostatin in its ability to damage DNA, the diol monomethyl ether derivative was prepared, and its stability and biological properties were studied. This analogue was found to be more stable than the epoxide (about 9-fold), but to be much less active in nicking supercoiled DNA and in forming covalent adducts with poly(dA-dT). However it is able to bind noncovalently to DNA and to the neocarzinostatin apoprotein. Another analogue, the chlorohydrin derivative, is about half as active as the epoxide in the DNA scission reaction and appears to produce the same covalent adducts with poly(dA dT) as does the epoxide, suggesting that both compounds undergo similar types of activation by thiol. These results indicate that the epoxide moiety of the neocarzinostatin chromophore is an important part of the highly unsaturated, strained bicyclo[7.3.0]dodecadiendiyne in the thiol-dependent, DNA-damaging reaction. It may be involved in the activation of the drug to its active species and/or may be the site of new bond formation in its reaction with DNA. PMID- 2965789 TI - Covalent labeling of vasopressin receptors from LLC-PK1 cells by the use of a bifunctional reagent. AB - The possibility of covalently attaching vasopressin to its receptors by the use of a bifunctional reagent was explored. Plasma membranes from the LLC-PK1 pig kidney cell line were purified by Percoll density gradient centrifugation. These membranes contained a single population of high affinity (Kd = 5.2 nM) and high capacity (Bmax 3.8 pmol/mg of protein) [3H]lysine vasopressin ([3H]LVP)-binding sites. [3H]LVP-labeled receptors could be solubilized with a high yield (83%) and minimal dissociation (9%) by treatment with the non-ionic detergent, octaethylene glycol mono-n-dodecyl ether (C12E8) (0.5%, v/v) in the presence of glycerol (20%). The solubilized [3H]LVP-labeled receptors were stable upon storage at 4 degrees (5% dissociation after 24 hr). They were partially purified to a specific activity of 17 pmol/mg of protein by chromatography on a Cibacron blue-Sepharose column with a yield of 90%. The [3H]LVP-receptor complexes in both intact membranes and the partially purified preparation were almost completely dissociated by incubation at 30 degrees for 30 min in the presence of 20 mM ethylenediaminetetraacetate (EDTA). This property was used to test the effect of ethylene glycol bis (succinimidyl-succinate) (EGS) as cross-linking reagent for the covalent attachment of [3H]LVP to its receptors. After treatment of [3H]LVP labeled membranes for 30 min with 1 mM EGS at 4 degrees, about 30% of specifically bound [3H]LVP was resistant to EDTA dissociation. The amount of EDTA resistant binding varied as a linear function of the fractional receptor occupancy and maximal binding capacity of the different batches of membranes used. Similar results were obtained with solubilized and partially purified vasopressin receptors. Upon steric exclusion high performance liquid chromatography, the EDTA-resistant [3H]LVP-labeled material, like the native [3H]LVP-labeled receptor, was eluted as a single and apparently homogeneous peak. The covalent character of the EGS-induced [3H]LVP binding to solubilized or partially purified receptors was assessed by its resistance to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The yield of EGS-induced labeling deduced from these experiments (27%) was close to that determined by the EDTA method. SDS-PAGE analysis of the [3H]LVP-labeled cross-linked material revealed the specific labeling of a major 50-kDa component and a minor component of 30 kDa. The size of these two components was not affected by dithiothreitol. PMID- 2965790 TI - Adenovirus E1A products suppress myogenic differentiation and inhibit transcription from muscle-specific promoters. AB - The primary function of the adenovirus E1A-region genes is to activate other adenoviral genes during a permissive viral infection by modifying the host cell transcriptional apparatus. Host cell immortalization, or transformation by the whole adenoviral early region, presumably results as a consequence of these modifications. Both transcriptional activation and transcriptional repression of non-adenoviral genes by the E1A proteins have been reported. It is currently not clear which, if either, of these activities contributes to host cell transformation and immortalization. Although there may be a physiological impact of some E1A-stimulated host cell genes, in many cases the functional significance is unclear. No common target sequences have been recognized in stimulated cellular genes and it has recently been proposed that in many cases, particularly involving newly transfected genes, available 'TATA-box' sequences may be the opportunistic beneficiaries of E1A assistance as a secondary consequence of E1A primary functions within the host cell nucleus. E1A-mediated transcriptional repression appears to be a more specific process insofar as common core elements are shared by the E1A-suppressed SV40, polyoma B, IgG heavy-chain and insulin enhancers. In the present communication we report that the complete myogenic programme of L8 and C2 myoblasts can be blocked by the introduction of constitutively expressing E1A genes, and show that the transcriptional induction of muscle-specific genes is inhibited. In particular, the promoter-inducing activities of well-defined elements that are required for the muscle-specific expression of the two sarcomeric alpha-actins, and which normally bind cellular trans-acting factors, become targets for E1A suppression. The results support the hypothesis that the suppression of differentiation by E1A products is effected by an E1A-mediated block in the transcriptional activation of cellular genes by specific developmentally regulated cis-acting promoter elements. PMID- 2965791 TI - Microtubule-associated protein 1C from brain is a two-headed cytosolic dynein. AB - Dynein, an ATPase, is the force-generating protein in cilia and flagella. It has long been speculated that cytoplasmic microtubules contain a related enzyme involved in cell division or in intracellular organelle transport. A 'cytoplasmic dynein' has been described in sea urchin eggs, but because the egg stockpiles precursors for both cytoplasmic and ciliary microtubules, the role of this enzyme in the cell has remained unresolved. We recently found that the microtubule associated protein (MAP) 1C (ref. 6) from brain is a microtubule-activated ATPase that produces force in the direction corresponding to retrograde organelle transport in the cell. MAP 1C has several similar properties to ciliary and flagellar dynein. Here we show directly, using scanning transmission electron microscopy, that MAP 1C is structurally equivalent to the ciliary and flagellar enzyme and is the long-sought cytoplasmic analogue of this enzyme. PMID- 2965792 TI - Localization of the binding site for the human high-affinity Fc receptor on IgG. AB - A major pathway in the clearance of pathogens involves the coating of the pathogen with specific antibodies, and the binding of the antibody Fc region to cell receptors. This can trigger engulfment of the pathogen by phagocytes or lysis by killer cells. By oligonucleotide site-directed mutagenesis we have engineered a single amino acid change in a mouse IgG2b antibody (Glu 235----Leu) which now enables the antibody to bind to the FcRI (high affinity) receptor on human monocytes with a 100-fold improvement in affinity. This indicates that Leu 235 is a major determinant in the binding of antibody to FcRI and that the receptor may interact directly with the region linking the CH2 domain to the hinge. Tailoring the affinity of antibodies for cell receptors could help dissect their role in clearing pathogen. PMID- 2965793 TI - Independent evolution of structural and coding regions in a Neurospora mitochondrial intron. AB - The discovery of intervening sequences (introns) in eukaryotic genes has raised questions about the origin and evolution of these sequences. Hypotheses concerning these topics usually consider the intron as a unit that could be lost or gained over time, or as a region within which recombination can occur to facilitate the production of new proteins by exon shuffling. Additional complexities are observed in introns of mitochondrial and chloroplast genes which contain secondary structures required for messenger RNA splicing and open-reading frames encoding proteins. Here we describe differences in the organization of protein-coding sequences in the intron of the mitochondrial ND1 gene in two closely related species of Neurospora. These differences show that intron sequences involved in secondary structure formation and in protein coding can evolve as physically distinct elements. Indeed, the secondary structure elements of the ND1 intron can contain two different coding sequences located at two different positions within the intron. PMID- 2965794 TI - Suppression of experimentally induced autoimmune encephalomyelitis by cytolytic T T cell interactions. AB - Down-regulatory phenomena have been described in several experimental models of tissue-specific, T-cell-mediated autoimmunity. For example, resistance to active induction of experimental autoimmune encephalomyelitis (EAE) can be induced by pretreating animals with non-pathogenic inocula of autoantigen or effector cells. Moreover, animals that have recovered from one EAE episode are resistant to subsequent induction of EAE. In some models, resistance to EAE has been transferred with immune cells to naive recipients. These experiments, which were based on transfers of unseparated immune cell populations, are difficult to interpret. Immune suppression circuits are known to be complex and involve various distinct cellular subsets. To further complicate the issue, resistance to EAE can be transferred not only by suppressor cells, but also by encephalitogenic effector cells injected in 'subclinical' doses. We describe now the isolation of homogeneous T lymphocyte lines from the spleens of Lewis rats that had recovered from T-cell-mediated EAE (tEAE) caused by the MBP-specific T cell line S1. These spleen-derived T line cells express the CD8 phenotype and specifically respond to determinants on the inducing S1 line, but not to the autoantigen MBP. Furthermore, the anti-S1 cells selectively lyse the encephalitogenic S1 T line in vitro and efficiently neutralize their encephalitogenic capacity in vivo. PMID- 2965795 TI - [Developments in 10 years of angioplasty of the coronary vessels]. PMID- 2965796 TI - [An unusual cutaneous reaction pattern as a side effect of penicillamine: elastosis perforans serpiginosa]. PMID- 2965797 TI - Glucocorticoids and head injury. A possible participation of lipocortin (lipomodulin) in actions of the steroid hormones. AB - The actions of glucocorticoids require the synthesis of new proteins in many, if not all, cases. One such protein that mimics the actions of glucocorticoids has been isolated and characterized as lipocortin. This protein can mimic the anti inflammatory activity of glucocorticoids by inhibiting phospholipase A2. The effect of glucocorticoids on various stages of brain and spinal cord injury may be partly, although not totally, explained by the activity of lipocortin. PMID- 2965798 TI - EEG changes following increased blood-brain barrier permeability under long-term immobilization stress in young rats. AB - A continuous 8 h of immobilization stress in conscious young rats increased the blood-brain barrier (BBB) permeability to 131I-sodium in 12 out of 14 brain regions studied. A flattening of electroencephalographic (EEG) activity was noted during this time period. The mean cerebral blood flow (CBF) was reduced by 17% (during this time period) but the regional flow reduction was not related to the regional increase in BBB permeability. On the other hand, a correlation was observed between increased plasma and brain 5-HT levels and increased BBB permeability. p-Chloro-phenylalanine (p-CPA) pretreatment has prevented the occurrence of increased BBB permeability, and the flattening of EEG activity as well as 5-HT levels in plasma and brain. These results suggest that the long-term immobilization stress induces causally related sequential events in rats: enhancement of circulating 5-HT, impairment of BBB, free access of 5-HT into the brain, and eventually flattening of EEG. PMID- 2965799 TI - Immunogenicity of a recombinant yeast-derived hepatitis B vaccine (Engerix B) in children. AB - Eighty-one children seronegative for markers of hepatitis B virus infection aged 0-10 years received 10 micrograms of a yeast-based hepatitis B vaccine (Engerix B) at 0, 1, and 6 months. Eighty subjects responded with a geometric mean titre of 7640 IU/L one month after the third dose. All seroconverters produced high level of antibodies regardless of age, race, or sex. It is concluded that Engerix B used in the dose and schedule described is highly immunogenic. PMID- 2965800 TI - Hepatitis B vaccines: use in childhood. PMID- 2965801 TI - Contact allergy to fungicide. PMID- 2965802 TI - Testosterone, androstenedione, dehydroepiandrosterone sulfate, and sex-hormone binding globulin in pregnant alcohol abusers. AB - We compared serum concentrations of total and free testosterone, androstenedione, dehydroepiandrosterone sulfate (DHEAS), and sex-hormone-binding globulin in 40 pregnant women exhibiting consistent alcohol abuse and in 20 abstinent pregnant controls. Sixteen drinkers gave birth to infants with fetal alcohol effects; the remaining 24 drinkers delivered healthy infants. Drinking patients with healthy infants had normal concentrations of total and free testosterone, sex-hormone binding globulin, and DHEAS, but their levels of androstenedione increased between weeks 20-40 of pregnancy. Maternal drinking leading to fetal alcohol effects was accompanied by lowered concentrations of sex-hormone-binding globulin throughout pregnancy and by low total testosterone concentrations, although the latter difference reached statistical significance only at 16-20 weeks' gestation. In contrast, drinking leading to fetal damage was associated with higher free testosterone levels between weeks 16-20 of pregnancy and lowered concentrations of DHEAS between weeks 16-32 of gestation, whereas the androstenedione levels tended to be high throughout pregnancy. These changes in androgens can be explained in part by low sex-hormone-binding globulin concentrations, an insufficient supply of DHEAS by the adrenals, and reduced peripheral conversion of androstenedione to testosterone. PMID- 2965803 TI - Low back pain presents tough challenge to occupational physicians. PMID- 2965804 TI - Pain patterns in adult scoliosis. AB - Adult patients with scoliosis often have back pain, but that pain may or may not be due to the curvature. A careful history, physical examination, routine radiographic examination, and, on some occasions, specialized radiographs, CT, myelography, discography, and facet joint injection will help the physician or surgeon separate out those pain syndromes owing to the curvature versus those not owing to the curvature. Only after these critical evaluations have been done can a decent decision be made as to the area of the spine to be treated, either surgically or nonsurgically. PMID- 2965805 TI - [Defining ATPase activity in functionally different parts of the body of the cestode Bothriocephalus scorpii]. AB - Data on the distribution of ATP-activity (A) along the longitudinal axis of the body of Bothriocephalus scorpii are given. A has been shown to decrease from the head to the posterior parts of the body. However, A has been noted to increase a little on the part of the body where mature eggs are thrown out. PMID- 2965806 TI - Life-saving therapy for newborns: a questionnaire survey in the state of Massachusetts. AB - The attitudes of Massachusetts pediatricians concerning life-saving therapies for newborns were surveyed. The sample was drawn from the membership list of the Massachusetts Chapter of the American Academy of Pediatrics. Of the sample population of 801 physicians, 449 (56%) responded. Three hypothetical cases were presented with treatment options: (1) An infant with Down syndrome and duodenal atresia, (2) an infant with meningomyelocele and microcephaly and, (3) an extremely low birth weight infant of 700 g and 28 weeks' gestation suffering from birth asphyxia. For case 1, 73% of the physicians recommended surgical correction of the duodenal atresia. Of these, 68% would seek a court order if the parents did not consent to surgery. For case 2, 53% of the physicians recommended surgical repair of the meningomyelocele. For case 3, 90% of the physicians recommended continued resuscitation of the infant and referral to an intensive care unit. Religious affiliation and age were factors influencing these clinical recommendations, with Catholic and younger physicians tending to opt for more aggressive medical intervention. The great majority of physicians (93%) indicated that the economic situation of the family should not affect a decision regarding life-saving therapy. A majority (62%) of the physicians approved of infant care review committees with a primary advisory role. Comparison with earlier attitudinal surveys suggests that pediatricians today are more willing to intervene medically on behalf of infant patients than previously. PMID- 2965807 TI - The interaction of Escherichia coli integration host factor with the cohesive end sites of phages lambda and 21. AB - The interaction of E. coli integration host factor (IHF) with the cohesive end sites (cos's) of phages lambda and 21 has been studied by the DNAase I footprinting technique. Six potential sites in cos lambda differ from the consensus IHF binding sequence by 1 to 3 base pairs. Of the six, one site, I1, binds IHF strongly. The I1 segment protected by IHF contains two sequences that closely match the IHF consensus binding sequence. Another site, I2, binds IHF moderately well, and three sites: 10', 13 and 14 bind IHF very weakly. The 10 site does not bind IHF under the conditions used here. In phage 21 the DNA segment extending to the right from the cohesive ends, which contains three potential IHF binding sites, was examined. Two sites bind IHF well; I1, the 21 analogue of one of the lambda I1 sites, and I0, a site not analogous to a lambda site. The third 21 site, I2, binds IHF moderately well, as does the analogous I2 site in lambda. The significance of the results for lambda DNA packaging is discussed. PMID- 2965808 TI - [Evaluation of cerebrovascular circulation in healthy persons using Doppler transcranial ultrasonography]. PMID- 2965809 TI - Shoe insoles in the workplace. AB - Ninety-six women participated in a crossover study to evaluate the effectiveness of viscoelastic polyurethane insoles in reducing back, leg, and foot pain among adults who spend the majority of each work day standing. Twenty-five of the subjects reported that the insoles made their shoes too tight to be comfortable. The remainder, however, found the insoles very comfortable (P less than .002, Wilcoxon, signed-rank test) and reported significant reductions in back pain (P less than .02), foot pain (P less than .03), and leg pain (P less than .007). When these subjects were asked whether they would prefer to wear their shoes alone or with insoles, the preference for insoles was overwhelming (P less than .007, back; P less than .03, leg; and P less than .009, foot pain). It is concluded that viscoelastic insoles can effectively improve comfort and reduce back, leg, and foot pain in individuals who must stand throughout the day. PMID- 2965810 TI - Comparison of desipramine, amitriptyline, zimeldine and alaproclate in six animal models used to investigate antidepressant drugs. AB - In the present paper the acute actions primarily of the tricyclic antidepressants amitriptyline and desipramine, the atypical antidepressant zimeldine and the potential antidepressant alaproclate were evaluated in six models used for studying antidepressant agents. These included the forced swim test, a modified learned helplessness procedure, the clonidine hypothermia test, the social dominance test (using the interaction with clonidine), a differential reinforcement-of-low-rates (DRL-72s) schedule and conditioned avoidance response. The results showed desipramine to be effective in all the tests employed. Zimeldine was effective in the learned helplessness, DRL-72s and domination tests, but also caused notable deficits in two-way active avoidance response. Alaproclate was effective in all the tests except the domination paradigm. Amitriptyline was effective in all tests employed. The results are discussed in relation to the possible mechanism of action of these compounds in the test models employed. PMID- 2965811 TI - A one year follow-up study of the distribution of 14C-clodronate in mice and rats. AB - The distribution of 14C-clodronate (dichloromethylene bisphosphonate) was studied in mice and rats by whole-body autoradiography and by measuring the 14C activities in various tissues up to one year after a single intravenous dose. The results indicate that the deposition of clodronate in bone tissue of mice and rats is extremely strong and long lasting; in both animals, high radioactivities were measured even 12 months after the administration. In spleen, thymus, and small intestine of mice, 14C-activity could be detected 12 months after dosing. Small amounts of radioactivity were also detected in spleen of rats for 12 months, although much less than in spleen of mice. The results suggest that clodronate accumulates to some extent in the reticuloendothelial system of animals. They suggest further than the animal species should be carefully considered when interpreting the distribution kinetics of bisphosphonates, especially their accumulation in the reticuloendothelial system. PMID- 2965812 TI - Purification and functional characterization of a cellular transcription factor that binds to an enhancer element within the adenovirus early EIIa promoter. AB - The adenovirus EIa-inducible early EIIa (EIIaE) promoter is comprised of several sequence elements essential for constitutive and induced expression. We report here the purification of the host-cell factor that interacts with the major upstream element of this promoter, extending between positions -90 and -70 with respect to the main EIIaE cap site and exhibiting enhancer properties. The purified factor, which corresponds to a 40- to 43-kDa polypeptide, specifically binds to its recognition site and stimulates EIIaE promoter activity when added to an in vitro transcription system, reconstituted from purified factors and RNA polymerase. The implication of this factor in the control of the other adenovirus early genes is discussed. PMID- 2965813 TI - NusA protein is necessary and sufficient in vitro for phage lambda N gene product to suppress a rho-independent terminator placed downstream of nutL. AB - Transcription antitermination by phage lambda N protein is reproduced in vitro solely with purified components. We have placed a strong rho-independent terminator, lambda tR', in the PL operon about 200 base pairs downstream from the N-recognition site, nutL, and have monitored terminated and run-off transcripts produced by single-round transcription of linear plasmids. In the presence of NusA, one of several host factors implicated in antitermination, N is found to virtually abolish termination at tR'. N is unable to suppress termination if the terminator is preceded by a defective nut site. Thus, during transcription through the nut site, N and NusA can modify RNA polymerase to a termination resistant form in the absence of any other accessory factor. PMID- 2965814 TI - Helper T-cell clones that recognize autologous insulin are stimulated in nonresponder mice by pork insulin. AB - Murine antibody responses to various species of insulin are under major histocompatibility complex-linked Ir gene control. Beef insulin differs from pork insulin by only two amino acids in the A-chain loop, yet strain C57BL/10 (B10) mice produce insulin-specific antibodies after immunization with beef insulin and fail to produce antibody after stimulation with pork insulin. Nevertheless, pork insulin primes helper T cells in B10 mice that can be demonstrated if insulin specific Lyt-1-, -2+ suppressor T cells are removed. Not only do the pork insulin primed helper and suppressor T cells cross-react with autologous insulin, but also rat insulin (the amino acid sequence of which is identical to mouse insulin) elicits functionally identical helper and suppressor T cells. In this report, we demonstrate that in B10 mice the frequency of helper T cells stimulated by pork insulin is equivalent to that stimulated by beef insulin and that helper T-cell clones induced by beef and pork insulin are major histocompatibility complex restricted T cells that proliferate, produce lymphokines, and provide helper activity after activation. These helper T-cell clones exhibit different antigenic fine specificities: beef insulin-induced clones respond to beef insulin but not pork or autologous insulin, whereas pork insulin-induced clones cross-react with all species of insulin tested, including rat insulin. In addition, the helper activity of cloned pork insulin-specific T cells is abrogated by pork insulin primed suppressor T cells. These data support the hypotheses that Ir gene control of antibody responses to certain antigens involves mechanisms used for maintenance of self-tolerance. PMID- 2965817 TI - Society of Cardiovascular and Interventional Radiology: credentials criteria for peripheral, renal, and visceral percutaneous transluminal angioplasty. PMID- 2965816 TI - Trisomy 21 in newborn infants: chest radiographic diagnosis. AB - Chest radiographs of 30 infants with trisomy 21 and 881 unaffected infants were evaluated for the following findings common in trisomy 21: multiple manubrial ossification centers, 11 rib pairs, and a bell-shaped chest. Radiographs were obtained in the first 48 hours of life. Of the 881 unaffected infants, 85 (9.6%) exhibited multiple manubrial ossification centers; 46 (5.2%), 11 rib pairs; and 208 (23.6%), bell-shaped chest. Of the 30 infants with trisomy 21, 24 (80%) exhibited multiple manubrial ossification centers; ten (33%), 11 rib pairs; and 24 (80%), bell-shaped chest. The probability of trisomy 21 in routinely radiographed newborn infants is 0.05% when none of the three findings is present, 1.6% with multiple manubrial ossification centers alone, 0.2% with 11 rib pairs alone, and 0.7% with bell-shaped chest alone. The probability of trisomy 21 increases in patients with two findings and reaches 58.4% in patients with all three findings. PMID- 2965815 TI - rpt-1, an intracellular protein from helper/inducer T cells that regulates gene expression of interleukin 2 receptor and human immunodeficiency virus type 1. AB - The Rpt-1 (for regulatory protein, T-lymphocyte, 1) gene, selectively expressed by resting but not by activated CD4+ inducer T cells, encodes an intracellular protein (rpt-1, Mr 41,000) that down-regulates gene expression directed by the promoter region of the gene encoding interleukin 2 receptor alpha chain and by the long terminal repeat of human immunodeficiency virus type 1. The data reported here suggest that rpt-1 levels may be inversely correlated with activation of CD4+ T cells and human immunodeficiency virus replication leading to clinical symptoms of the acquired immunodeficiency syndrome. PMID- 2965818 TI - Laser angioplasty: controlled delivery of argon laser energy. AB - A laser catheter system that integrates balloon and fiberoptic technologies was evaluated. In vitro tissue studies were conducted to compare the tissue response to laser irradiation from a bare optical fiber (undiverged light beam) and from a fiber with an optical assembly (diverged light beam). An in vivo study of occluded and unoccluded canine femoral arteries examined coaxial alignment of the diverged light beam and the resultant thermal effects. In the in vitro studies, a diverged laser light beam effected maximum tissue ablation (mean crater diameter, 1.81 mm +/- 0.44) at a distance of only 3 mm from the optical fiber tip. The in vivo study demonstrated the ability of a specially designed balloon catheter to align the diverged laser beam coaxially within the arterial lumen. This laser catheter system successfully avoids the risks of inadequate tissue removal and perforation. PMID- 2965819 TI - Legal case briefs for nurses: N.Y.: nurse seeking doctorate: discharged; R.I.: failure to report injury: workers' compensation. PMID- 2965820 TI - Molecular mechanisms involved in the desensitization of dopamine receptors in slices of corpus striatum. PMID- 2965821 TI - Phosphoprotein B-50: localization and function. PMID- 2965823 TI - [The peak flow meter]. PMID- 2965822 TI - Comparison between the neural acidic proteins B-50 and F1. PMID- 2965824 TI - [Anatomical study of the cutaneous innervation of the lumbosacral region. Application to the physiopathology of certain lumbalgias]. AB - 30 dissections have confirmed that the skin of the gluteal area is innervated by the posterior branches of T12 and L1, 19 times out of 30 (64 p. cent) or T12, L1 and L2, in 8 instances (26 p. cent) or T12, L1 and L2 receiving an anastomosis from L3, in 3 instances. This explains certain lumbo-gluteal pain coming from the dorso-lumbar area ("low lumbalgias of high origin"). In addition, the posterior branch, the most medial (most of the time L1, sometimes L2) goes through an osteo aponeurotic channel formed by the iliac crest below and the dorso-lumbar fascia above. This channel, located at 7 or 8 cm of the spinal process, may be at the origin of a neuropathy. It projects at the same level in relation to the mid-line as the iliac insertion of the ilio-lumbar ligament. The latter seems impossible to feel as it is too deep and hidden by the superficial aspect of the iliac crest. Lumbalgias, sometimes related to a strain of this ligament, seem to be, most of the time, the result of an irritation of the posterior branch of L1 or L2. PMID- 2965825 TI - [Long-term results of chymopapain nucleolysis]. AB - The retrospective study of the first 67 patients treated by chemonucleolysis, since December 1980, shows a good overall result, in 69 p. cent of the cases (40 patients) and a failure in 31 p. cent of the cases (18 patients), with a 42 months follow-up. The radiculalgia is cured or quite improved in one month in 90 p. cent of the cases and in 2 years in 88 p. cent. Lumbalgia is absent or moderate after 1 month in 58 p. cent of the cases and after 2 years in 81 p. cent. It persists, most of the time, in women, and this is statistically significant. 15 immediate failures and 3 recurrent sciaticas at the same level are noticed after a 22 months free interval. The possibility of severe complications requires a perfect technique and well-defined indications. The study confirms the good results of nucleolysis with a 42 months follow-up. PMID- 2965826 TI - Increased atrial natriuretic peptide in the nephrotic syndrome. Relationship to the renal function and the renin-angiotensin-aldosterone system. AB - Atrial natriuretic peptide (ANP), angiotensin II (Ang II), and aldosterone (Aldo) in plasma and creatinine clearance (Ccr) were determined during basal conditions in 17 patients with the nephrotic syndrome and 20 control subjects. In addition, six of the patients were studied after seven remissions of the syndrome. In the nephrotic syndrome ANP was higher than in the control group (9.7 (median) versus 7.2 pmol/l, p less than 0.01), Ccr was lower (55 versus 99 ml/min, p less than 0.01). Angiotensin II and Aldo were the same in patients and control subjects. After remission of the syndrome ANP was reduced (11.2 to 5.4 pmol/l, n = 7, p less than 0.02) and Ccr increased (52 to 84 ml/min, n = 7, p less than 0.02), whereas Ang II and Aldo were unchanged. A significant, negative correlation was found between ANP and Ccr in the subgroup of patients in whom the syndrome remitted (Q = -0.547, n = 14, p less than 0.05). Atrial natriuretic peptide was not correlated to either Ang II or Aldo in either of the groups. It is concluded that patients with the nephrotic syndrome have elevated ANP, and it is suggested that a high ANP may be a compensatory phenomenon induced by a decreased renal ability to eliminate sodium and water. PMID- 2965828 TI - [Hepatitis B screening in late pregnancy and results of immunization in newborn infants]. AB - Screening for hepatitis Bs antigen in late pregnancy was introduced in mid-1983 at the University Women's Hospital, Berne. 4118 pregnant women had been investigated by the end of 1986, of whom 26 (0.63%) turned out to be HBsAg positive. The prevalence of HBsAg carriers was 0.12% in Swiss women, 0.65% in other European women, 12.5% in women from the Far East and 5.6% and 4.9% in women from the Near East and Africa respectively. Newborns of HBsAg positive mothers simultaneously received hepatitis B immunoglobulin (400 IU anti-HBs) and a first injection of hepatitis B vaccine (Hevac B 5 mg) in the first hours of life, followed by vaccinations after two, three and twelve months. Of 18 children investigated after the first year of life none was HBsAg positive. 14 children (78%) were shown to have HBs antibodies. Two of the four anti-HBs-negative unfortunately received only the first vaccine injection after birth. Taking this fact into account, the "success rate" of the immunoprophylaxis is 88%. PMID- 2965827 TI - [Treatment of lumbar sciatica with or without neurological deficit using mechanical traction. A double-blind study]. AB - Traction therapy for low back pain with sciatica has been evaluated in a double blind study. 60 patients hospitalized for sciatica with or without signs of sensory or motor deficiency were randomized to 3 treatment groups: "placebo traction" (5 kg), "light traction" (15 kg) and "normal traction" (50 kg). Clinical evaluation after 4, 8 and 12 traction sessions showed no difference between the three groups. PMID- 2965829 TI - Injury to the abdominal wall musculature: the full spectrum of traumatic hernia. AB - It is unusual for all anterior abdominal wall muscles to be disrupted in blunt trauma. We have described such a case, which necessitated reevaluation of the recommendations for treatment of traumatic hernia. We have proposed a method of classification of the different types of traumatic hernias so that treatment can be tailored. PMID- 2965830 TI - Huntington disease in South African blacks. A report of 8 cases. AB - Huntington chorea is rare in ethnically pure blacks; 8 South African black patients, 6 adults and 2 children, from 4 families are described. The symptoms do not appear to differ materially from Huntington chorea in other races. Increased awareness of the disease in this population group is warranted. PMID- 2965831 TI - [Comparative evaluation of the structural-functional state of the renal arteries in health and disease using combined two-dimensional Doppler echography]. PMID- 2965833 TI - [Current broncholytic and expectorant therapy of patients with chronic nonspecific lung diseases]. AB - Correction of the obstructive syndrome in patients with chronic nonspecific pulmonary diseases using modern broncholytics and expectorants is considered. The chief mechanisms of the formation of the obstructive syndrome (bronchospastic and obturation ones as a result of mucociliary transport disorder) are described. The authors propose a scheme of differential clinical and laboratory diagnosis of the above mechanisms of bronchial permeability disorder. Modern broncholytics and expectorants are characterized, mechanisms of their action and practical aspects of their application in chronic nonspecific pulmonary diseases are discussed. PMID- 2965832 TI - [Preasthma and bronchial asthma: problems of clinical immunology]. AB - The paper is concerned with the results of a study of the nature of immunological disorders in 150 patients with chronic bronchitis and preasthma and in 203 patients with bronchial asthma (BA). Subpopulation analysis of immunocompetent cells with the help of monoclonal antibodies permitted the detection of certain differences in the regulatory link of immunity in patients with different types of chronic nonspecific pulmonary diseases. Patients with purulent bronchitis demonstrated a decrease in the level of the population of T-helpers and a tendency to an increase in the level of T-suppressors, whereas preasthmatic patients demonstrated a decrease in the level of T-helpers in parallel with a sharp deficiency of T-suppressors. An analysis of immune disorders in BA patients revealed the same line of immunological shifts as in preasthmatic patients, however a degree of the affection of the suppressor systems was much greater. The study has shown that immune disorders typical of BA, are revealed even at the early stages of disease, i.e. at the preasthmatic stage. Possible immune mechanisms of pathogenesis of chronic bronchitis, preasthma and BA are discussed. A conclusion is made that immunological investigation including the determination of the levels of T-helpers and T-suppressors and their ratios (the so-called immunoregulatory index) in combination with other methods makes it possible to detect persons at risk of developing BA, at the early stages of development of disease. Immunocorrective therapy is recommended to such patients to prevent development of chronic bronchitis into BA. PMID- 2965834 TI - Transient inhibition of mixed lymphocyte reactivity by dieldrin in mice. AB - Dieldrin, a non-aromatic organochlorinated pesticide, was shown to be a potent modulator of the immune system. We had earlier demonstrated that mice treated with a single sublethal dose of dieldrin showed an impaired antibody response and reduced viral restriction mediated by macrophages. These dieldrin-induced immunosuppressive effects were shown to be dose dependent, when administered by the oral or intraperitoneal (i.p.) route. This study was undertaken to investigate the effect of dieldrin on the T-cell immune response. Lymphoid cells from mice injected i.p. 7 days earlier with 36 mg/kg body weight (0.6 LD50) dieldrin were assessed for their ability to recognize a foreign antigen and to proliferate in a mixed lymphocyte reaction (MLR) at 4, 7, 14 and 24 days post treatment. We have demonstrated a strong but transient inhibition of MLR at 7 days after pesticide exposure. This effect was reversible and could not be attributed to a direct cell cytotoxicity, nor to the modulation of the T-cell ratio in lymphoid organs. Since the mitogen response was not impaired at this time point, we suggest that T-cell ability to recognize a foreign antigen can be altered by dieldrin, but not the proliferative potential of the cells. PMID- 2965835 TI - Evidence that IFN-gamma is responsible for natural suppressor activity in GVHD spleen and normal bone marrow. AB - Natural suppressor (NS) cells are capable of suppressing immunological responses in a nonspecific manner. Previously, we have described NS cells in the spleens of mice undergoing chronic graft-versus-host disease (GVHD) and also in normal B10.D2 bone marrow (BM). NS cells obtained from these environments appear dependent upon lymphokines for their ability to manifest suppression. In this report, with anti-IFN-gamma antibody, we show that IFN-gamma is necessary for NS cell activation. Anti-IFN-gamma antibody is able to remove the ability of NS cells to suppress a concanavalin A (Con A) proliferation assay. Also, anti-IFN gamma antibody removes the ability of rIL-2, lectin-free Con A supernate (CAS), and recombinant IFN-gamma (rIFN-gamma) to enhance NS suppression of lipopolysaccharide response. By these criteria, IFN-gamma is required for NS cell activation, and rIL-2 may act indirectly by its ability to stimulate IFN-gamma synthesis. These results are discussed in the context of the immuno-suppression seen in human BM transplantation. PMID- 2965836 TI - Effect of blood transfusions from different H-2 donors on immune responses in mice. AB - We studied the effect of blood transfusions (BT) from different H-2 donors on the induction of suppressor cells (SC) and of MLC inhibitory activity in serum in a drug-unmodified mouse model. Balb/c (H-2d) mice were transfused at weekly intervals with whole blood from donors of three strains using two transfusion protocols. In protocol I, blood was transfused first from C3H/HeJ (C3H) (H-2k), then C57Bl (H-2b), and then SJL (H-2s) strain mice, and in protocol II the order of blood donors was reversed. Spleen cells and serum samples were obtained from the transfused mice one and two weeks after the last BT. In both transfusion protocols, the kinetics of responses of cells from recipient transfused mice to cells from the blood donors in MLC were similar to those of cells from nontransfused mice. The peak responses of cells from transfused mice were consistently lower than those of cells from nontransfused mice. In cell-mixing experiments, radiosensitive SC capable of inhibiting responses of Balb/c mice to cells from all three blood donors in MLC could be demonstrated one week after the last transfusion in both protocols. Two weeks after the last BT, SC were demonstrable only against the first (C3H) blood donor in protocol I, and against all three blood donors in protocol II. Serum obtained one week after transfusion in protocol I inhibited responses of Balb/c mice to stimulator lymphocytes from all three blood donors in MLC. Serum obtained two weeks after BT, however, inhibited responses of recipient mice only to the first blood donor. In contrast, in protocol II, serum obtained both one and two weeks after BT did not cause inhibition of responses of cells from Balb/c mice to blood donor cells in MLC. Similar results were obtained when Balb/c mice were transfused at weekly intervals with whole blood from either C3H or from SJL mice. The data suggest that the induction of SC and/or MLC-inhibitory activity in the serum after BT is dependent on the H-2 type of the first blood donor. PMID- 2965837 TI - Mixed lymphocyte culture--evidence that pretransplant transfusion with platelets induces FcR and blocking antibody production similar to that induced by leukocyte transfusion. PMID- 2965839 TI - Controlling lungworm. PMID- 2965840 TI - [Unna-Thost keratoderma palmaris et plantaris in 3 generations]. PMID- 2965838 TI - [Qualitative and quantitative changes in the hypothalamus and the ependyma of the 3d cerebral ventricle in sheep after irradiation and hormonal stimulation]. AB - In the submitted work we concentrated our attention on the study of the changes in neurosecretion. PAS positive mucopolysaccharides in the hypothalamus and ependyma of the IIIrd cerebral ventricle and on studying the changes in the volume of the cell nuclei of the neurons of nucl. supraopticus (NSO), nucl. hypothalamicus centromedialis (NVM) and nucl. tuberomamillaris (NTM) of sheep after irradiation and after hormonal stimulation. We made our observations on 28 sheep of the Slovak Merino breed, of average live weight 35 to 40 kg, two- and three-years old in the period of physiological anoestrum. The first group of four ewes and the second group of four yearling lambs were controls. The third group of six ewes and the fourth group of six yearlings were exposed to irradiation all over with a dosage of 2.46 Gy (250 R) for a period of five days. To synchronize the oestrum of all the sheep we used agelline sponges which we fitted five days before irradiation. After the irradiation and removal of the sponges we gave the test sheep hormonal stimulation with 3 X 500 i. u. of serum gonadotropin. The fifth group of four ewes and the sixth group of four yearlings were only stimulated without irradiation. The fifth day after stimulation we slaughtered the sheep. We processed the brain samples by the usual histological methods. We carried out karyometric analysis with 3000X magnification and the measurement of 200 cells from one sample. We processed the values obtained mathematically according to Fischer and Inke (1956). We evaluated the quantity of neurosecretion material with a light microscope according to Nakahara (1962). We found that the multiplication of neurosecretion and the increased activity of the PAS reaction in the hypothalamic nuclei studied show that the irradiation and hormonal treatment stimulate the function of the hypothalamic structures. The results of the karyometric analysis in the yearlings also confirm this opinion. In the ewes the inhibitive influence of irradiation was probably manifest. The giving of hormones in combination with irradiation causes the multiplication of ependyma cells and the desquamation of surface layers. PMID- 2965841 TI - Comparison of enzyme-linked immunosorbent assay and indirect fluorescent antibody test for the detection of IgG antibodies to Sarcocystis muris. AB - Sarcocystis muris cystozoites were separated from host tissue after mechanical mincing and homogenization by discontinuous density gradient centrifugation using Percoll. The isolated antigen, consisting of more than 95% cystozoites, was used in an enzyme-linked immunosorbent assay (ELISA) and an indirect fluorescent antibody test (IFAT). In the ELISA, the antigen was used at a very low protein concentration of 0.44 micrograms/ml. Good reproducibility of test results was achieved with different conjugates and different reference sera when the measured optical densities were converted into an index related to a standard. From the examination of 500 sera from non-infected mice, and index of 0.300 at a serum dilution of 1:10 was determined as the threshold for a positive reaction in the ELISA. Specific antibodies were first detected between 18 and 49 days after experimental infection (dpi) with S. muris. The ELISA as well as the IFAT was highly sensitive from about 50 dpi onwards and detected antibodies up to the end of the examination period (182 dpi). However, the IFAT with unfixed antigen appeared more sensitive than the ELISA for the period of infection before 50 dpi and detected specific antibodies between 11 and 25 dpi starting with fluorescence at the apical pole of the cystozoite and resulting in bright fluorescence of the whole cystozoite from 32 or 35 dpi onwards. Both serotests showed only slight crossreactions with high titred Toxoplasma gondii sera at a serum dilution of 1:10. The activity of the antigen lasted for at least 13 months in the ELISA and for at least eight months in the IFAT. PMID- 2965842 TI - [Use of transcranial electroanalgesia for treating spondylogenic pain syndromes]. AB - The analgesic effect of the net transcutaneous application to the brain of constant and impulse currents (the frequency of rectangular impulses was 77 Hz, the duration of an impulse, 3.5 ms) in a 2-5:1 ratio was investigated. An analgesic effect was observed in both healthy subjects and in patients with spondylogenic pain. The duration of analgesic effect following a 30-minute therapeutic session was 6-10 hours. A 5-7 session course was associated with a gradual increase and stabilization of the analgesic effect. The procedure was attended by no complications. PMID- 2965843 TI - [Migration of mental patients due to disease-related motives]. AB - The authors have studied a group of mental patients who have migrated to Moscow from various administrative areas of the Russian Federation. It has been established that the number of such patients over the last ten years has decreased by 11.6 per cent only. The authors present data on the nosological composition of the group of migrants and mental disturbances in each nosological group with which the migrational behaviour is associated. It has been demonstrated that the number of migrants from variable areas diminishes with a greater distance from Moscow; it is lower in those areas where the outpatient care is better exercised. The ways of preventing the migrational behaviour of mental patients have been outlined. PMID- 2965844 TI - Pathology of diffuse lesion of the lymph node with cellular atypism--a proposal for diffuse lymphoid dysplasia or lymphodysplastic syndrome. PMID- 2965846 TI - Angioimmunoblastic lymphadenopathy with dysproteinemia, its position in T cell proliferative disorders. PMID- 2965845 TI - Proliferating cell population in angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) lesions analyzed by double immunoenzymatic staining and immunomolecular markers. PMID- 2965847 TI - Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma: clinico-pathologic, immunophenotypic and immunogenotypic analyses. PMID- 2965848 TI - Suppressive effect of quinolinic acid and hippuric acid on bone marrow erythroid growth and lymphocyte blast formation in uremia. PMID- 2965849 TI - Estimation of Newcastle disease virus concentration by using a sandwich ELISA and measuring neuraminidase activity. PMID- 2965850 TI - Laser thermal angioplasty for the treatment of obstruction of the distal superficial femoral or popliteal arteries. AB - The success rates and clinical effectiveness of percutaneous laser thermal angioplasty for obstruction of the distal superficial femoral or popliteal arteries were assessed. Laser thermal angioplasty was performed with the first commercially available laser system on 25 atherosclerotic vascular lesions in 20 lower extremities of 18 patients. Fifteen of the lesions were occlusions (average length, 5.6 +/- 3.1 cm), and 10 were high-grade stenoses (average length, 2.6 +/- 1.8 cm). Indication for angioplasty was claudication limiting life-style in 12 extremities and threatened limb loss in eight. All patients were treated with a 1.5- or 2.0-mm metal-capped optical fiber attached to a 14-W argon-ion laser. Balloon angioplasty was performed after laser recanalization in all successful cases. The patients were followed by Doppler ankle/brachial index and clinical evaluation. Laser thermal angioplasty was technically successful (vessel recanalization) in 18 (90%) of 20 of the cases. Fourteen (70%) of 20 extremities were successes by ankle/brachial index follow-up, while 15 (75%) of 20 remained clinically improved after an average follow-up of 7.8 +/- 5.3 months. Laser thermal angioplasty has a technical and clinical success rate that compares favorably with previous reports of balloon angioplasty. With proper patient selection, laser thermal angioplasty can reduce the difficulty of traversing lesions that are amenable to standard balloon angioplasty and increase the number of vascular lesions that can be treated percutaneously but could not be recanalized without the use of laser energy. PMID- 2965851 TI - Role of the intimal flap in arterial dissection: sonographic demonstration. PMID- 2965852 TI - Coronary artery laser therapy. PMID- 2965853 TI - A practical approach to workers' compensation. AB - An understanding of workers' compensation and of the physician's role in it can significantly improve communication with patients, insurance companies and employers. Early rehabilitation is essential. Treatment of difficult patients can be enhanced by recognizing and treating the factors that are delaying recovery, encouraging activity and minimizing the use of narcotics. Factors delaying recovery include depression, alcoholism and pending litigation against the employer. PMID- 2965854 TI - Identification of high-risk patients with silent myocardial ischemia after percutaneous transluminal coronary angioplasty by multilead monitoring. AB - Quantification of total ischemic burden can be improved by use of microprocessor driven, multichannel, digital, ST-monitoring devices. ST deviation recorded during angioplasty provides a patient-specific template for identification of patients at increased risk for coronary events after angiographically successful dilation. In patients with ST-segment elevation, evidence of these ST "fingerprint" patterns are associated with elevated risk for coronary events in the postangioplasty period. Such risk is not seen in patients with ST-segment depression or with no ST changes. The increased monitoring capability afforded with digitized 3- or 12-lead monitoring creates new criteria for what constitutes accurate determination of total ischemic burden. PMID- 2965856 TI - Phase II trial of menogarol in the treatment of advanced adenocarcinoma of the pancreas. AB - Fifteen patients with advanced adenocarcinoma of the pancreas were treated with menogarol 150-225 mg/m2 i.v. every 3 weeks. All patients had bidimensionally measurable disease. This regimen and dosage schedule are well tolerated, with minimal toxicity that included myelosupression; median white blood cell (WBC) count nadir of 2,700 cells/mm3 (range 1,400-7,100 cells/mm3) and median platelet nadir of 162,000 cells/mm3 (range 53,000-390,000 cells/mm3). Anorexia occurred in one patient, nausea or vomiting in six, phlebitis in one, and alopecia in six patients. No patients responded. At this dosage and schedule, there is no role for menogarol in the treatment of advanced pancreatic adenocarcinoma. PMID- 2965855 TI - Endorphins and pain perception in silent myocardial ischemia. AB - Evidence suggests that endogenous opioids, particularly the beta-endorphins and met-enkephalins, are closely involved in stress-induced analgesia and nociceptive pain control. Numerous investigations have been conducted to evaluate the role of opioids in silent vs symptomatic myocardial disease. There is good evidence to suggest that patients with asymptomatic ischemia have defective pain perception compared with those with angina; however, the precise role of the endorphin and enkephalin systems in this phenomenon remains to be elucidated. Possible sources for disparate study results include variation in patient populations, insensitive or improperly timed assay techniques, and differences in amount of ischemia. PMID- 2965857 TI - Ultrastructural studies of peripheral blood of neonates with Down's syndrome and transient abnormal myelopoiesis. AB - Ultrastructural studies were performed on blood samples from five neonates with Down's syndrome and transient abnormal myelopoiesis (TAM). Three methods of fixation were employed to detect diaminobenzidine (DAB) reactivity. The first method used glutaraldehyde solution, while the second and third methods used tannic acid-glutaraldehyde mixtures. Before fixation by the second method, specimens were washed in order to eliminate plasma, and before fixation by the third were diluted to lessen the effects of plasma protein. The latter two methods were more sensitive than the first for detection of DAB reactivity, while the third also resulted in better preservation of morphology than did the second. Even the first method was able to detect DAB reactivity in cells of megakaryocyte platelet series in appropriate sections. Although the majority of blasts appeared with light microscopy to be undifferentiated, their ultrastructural morphology and ultrastructural cytochemistry were in fact found to be quite heterogeneous, consisting of cells of the megakaryocyte-platelet and granulocytic series, including basophils, and erythroid precursors. This finding supported the view that TAM was the result of unstable hematopoiesis rather than true leukemia. PMID- 2965858 TI - Infantile glaucoma in Down's syndrome (trisomy 21). AB - We examined five patients with Down's syndrome and bilateral infantile glaucoma. In the first few months of life four patients had large cloudy corneas, breaks in Descemet's membrane, increased intraocular pressure, photophobia, and tearing. In one patient the diagnosis was delayed until 3 1/2 years of age because of concomitant nasolacrimal duct obstruction. Two patients developed cataracts and retinal detachment and have undergone multiple surgical procedures. The clinical course in these two older patients suggests that coexistence of congenital glaucoma, severe myopia, and cataracts in patients with trisomy 21 strongly predisposes for the development of retinal detachment and poor visual outcome. PMID- 2965859 TI - Studies on crown ether based potentiometric sensors for 4,4'-dipyridinium and related dications. PMID- 2965860 TI - [Arterial hypertension of renovascular origin]. AB - We present 5 renal artery stenosis in 3 patients, 2 of them affected of neurofibromatosis. Percutaneous transluminal angioplasty was successful in one left renal artery. The operation was carried out in 4 renal arteries, 2 aortorenal reimplantations, 1 splenorenal bypass and 1 patch angioplasty using saphenous vein were performed. At this time 2 children are normotensive and do not require medication, and 1 child is normotensive and is receiving antihypertensive medication at a smaller doses that those necessary before operation. The good permeability of anastomosis was verified in all cases. PMID- 2965861 TI - [Partial 21q de novo duplication. Case report]. PMID- 2965862 TI - [Down's syndrome with Turner mosaicism/Y chromosome in a ring: 46,X+21/47X(r)Y,+21]. PMID- 2965863 TI - Chlorozotocin-induced selection of autocrine and multidrug resistant variants from a rat rhabdomyosarcoma. AB - The ability of tumor cells to develop simultaneous resistance to structurally different cytotoxic drugs constitutes a major problem in cancer chemotherapy. We previously described that a nitrosourea, chlorozotocin (CZT), increased in vivo the pulmonary metastatic dissemination of a rat rhabdomyosarcoma and enhanced in vitro the cloning efficiency (CE) of the same tumor cell line. The selection procedure for chlorozotocin-selected cell lines (R1 to R9 lines), by in vitro CZT treatment and cloning assay, indicated that 2.5% of the parental cell line was a CZT-resistant subpopulation. This subset acquired, in the course of the selection steps in semi solid medium, a multidrug-resistant phenotype. In a stem cell assay, the resistance of the R9 line was increased 10-fold for adriamycin (ADR) and up to 2 fold for cisplatinum. In comparison, C8 control line sensitivity to ADR did not change significantly. This resistance did not affect the non clonogenic population, as shown in a monolayer proliferation assay. Re-exposure of R (R1-R9) lines to CZT or initial CZT contact with C lines transiently but consistently increased the CE; however, clonogenicity remained stable, and this was also observed when deticene, another alkylating drug, was used instead of CZT. Moreover, the CZT-selected R9 line easily proliferated in serum-free medium in the presence of transferrin and insulin, whereas serum was necessary to maintain the growth of the parental cell line or C lines. Finally, we show that in vitro selection of variants, by both resistance to CZT and cloning, led to the isolation of a multi-drug resistant subpopulation, serum independent for its proliferation - two properties associated with progressive malignancy. PMID- 2965865 TI - Selected pharmacological and immunopharmacological indices of ibuprofen. AB - Affinity of ibuprofen to cellular immune reactions was evaluated in the light of its basic pharmacological indices. Ibuprofen was observed to modulate some immunological phenomena with marked predominance of immunosuppressive component. It has been shown that at least to some extent, mechanism of its effect consists in an interference in T-suppressor cell activity. PMID- 2965864 TI - Growth inhibitory effect of LH-RH analogs on human breast cancer cells. AB - The antiproliferative effect of two LH-RH agonists (Pro 9-LH-RH ethylamide and D Ser(t BU)6 Aza Gly 10-LH-RH, ICI 118630) on the human breast cancer cell line CG5 is reported. Although ineffective when used alone, both analogs inhibited in a dose-dependent fashion the growth stimulatory effect of estradiol. LH-RH analogs did not influence the growth inhibitory effect of tamoxifen and medroxyprogesterone acetate. Likewise, these compounds neither modified basal estrogen and progesterone receptor levels nor prevented estrogen-induced increase of progesterone receptor. A marked antiproliferative effect of the analogs was also seen in cells stimulated with other mitogens, such as insulin and epidermal growth factor. PMID- 2965866 TI - Immunostaining for the leukocyte common antigen. PMID- 2965867 TI - The CaATPase activity of rat-incisor odontoblast vesicles. AB - An homogenate of rat incisor odontoblasts had Ca2+ and Mg2+ ATPase activity and suitable storage conditions kept it stable for several days. Over 90 per cent of the activity was retained in a vesicle-rich microsomal fraction that removed about 85 per cent of the total material from the homogenate. This fraction was further characterized: the resolved Ca2+-activated ATPase activity, above the basal MgATPase activity, was 0.30 mumol Pi/min-mg total protein, and 50 per cent activated at free [Ca2+] equal 0.8 microM. This calcium dependency is consistent with an intracellular Ca2+-regulated enzymatic activity. The calcium ionophore, A23187, had no measurable effect on the CaATPase activity, which suggests that the odontoblast vesicles do not concentrate Ca2+ in a lipid bilayer compartment. Direct measurement of the uptake of 45Ca2+ by the filtration method and parallel measurements of CaATPase activity on the same preparations under identical conditions indicated that the odontoblast-derived vesicles have a coupling ratio of 0.024 Ca2+/ATP. This low coupling ratio and the lack of detectable compartmentalization of calcium indicate that the CaATPase activity of the odontoblast microsomes is not associated with a calcium pump. The [Ca2+] dependence of the activity suggests the CaATPase is under intracellular Ca2+ control, but its function is unknown. PMID- 2965868 TI - Thoracic back pain. The thigh extension technique. PMID- 2965869 TI - Chlorambucil allergy--a series of three cases. AB - Three cases of chlorambucil-related skin eruption in patients with low-grade non Hodgkin's lymphoma are described. Cases were confirmed by exclusion of other drugs, rechallenge, skin testing and histology. One patient died due to florid erythema multiforme. Only two other adequately documented cases are described. PMID- 2965870 TI - The internal crosslinking of the S1 heavy chain from smooth muscle probed by dibromobimane. AB - The reaction of the crosslinker dibromobimane has recently revealed a functionally important internal loop structure within the skeletal myosin S1 heavy chain where Cys-522 of the 50K domain and Cys-707 (SH1) of the 20K region are spatially juxtaposed. Here we have studied the possible relevance of this topological feature to the architecture and transducing activity of the myosin head in general, by extending the dibromobimane modification to smooth muscle myosin. Treatment of chicken gizzard myosin S1 with dibromobimane resulted in intramolecular crosslinking between the C-terminal 25K and central 50K segments of the S1 heavy chain. The data suggest a conservation at the 50K-25K interface of smooth muscle S1 heavy chain and the importance of the neighboring SH1 region, whose conformation may play an important role in energy transduction by the myosin head. PMID- 2965871 TI - Degradation of [125I]-atrial natriuretic peptide by a soluble metallopeptidase isolated from rat ventricular myocytes. AB - Atrial natriuretic peptide is rapidly degraded by a soluble, heat labile peptidase isolated from ventricular myocytes. Degradation of [125I]-ANP is antagonized by unlabelled ANP, bradykinin, glucagon, 1,10-phenanthroline, PCMB, EDTA and the bacterial antibiotic bacitracin, but not by phenylmethylsulphonyl fluoride, aprotinin, phosphoramidon, E-64, amastatin or the ACE inhibitor SQ 20881 and bradykinin potentiator C. In addition neither bovine serum albumin nor caesin afforded any protection against degradation. Peptidase activity was optimal at pH values above 8.5. The peptidase is likely to be of intracellular origin and may contribute to the extensive ANP degradative activity found in various ventricular muscle preparations. PMID- 2965872 TI - Interleukin-one induced inositol phospholipid breakdown in murine macrophages: possible mechanism of receptor activation. AB - Stimulation of mouse peritoneal macrophages by Interleukin-one (IL-1) provoked rapid increases in the levels of inositol mono, bis, tris and tetrakisphosphates (IP1, IP2, IP3 and IP4 respectively). IP3 was by far the major metabolite formed and time course studies revealed that IP2 and IP3 were formed more rapidly than IP1 and IP4 in response to IL-1 stimulation. The IP2 and IP3 levels peaked at five seconds while there was a time lag of five seconds in the IP4 response and the IP1 levels increased relatively steadily over the time course of the experiments. Levels of IP2, IP3 and IP4 all returned almost to control levels by 60s. The rapid formation of the inositol phosphate metabolites was concomitant with a decrease (84%) in the levels of phosphatidyl-inositol 4,5-bisphosphate (PIP2) in the macrophages. These results suggest that the mechanism of IL-1 receptor activation is by the rapid hydrolysis of phosphoinositides and generation of the second messenger IP3. PMID- 2965873 TI - Impaired proteolytic processing of lysosomal N-acetyl-beta-hexosaminidase in cultured fibroblasts from patients with infantile generalized N-acetylneuraminic acid storage disease. AB - Cultured skin fibroblasts from patients suffering with infantile generalized N acetylneuraminic acid (NeuAc) storage disease accumulate free NeuAc in a population of lysosomes less dense than those observed in normal fibroblasts (1.035 vs. greater than 1.07 mean density), as assessed by the distribution of lysosomal enzyme activities and NeuAc on Percoll gradients after subcellular fractionation. In the present study, normal and affected fibroblasts were labeled with [35S]methionine, and cell homogenates or subcellular fractions from Percoll gradients were immunoprecipitated with polyclonal antibodies to lysosomal N acetyl-beta-hexosaminidase (Hex); immunoprecipitated polypeptides were analyzed by SDS-polyacrylamide gel electrophoresis. The synthesis and initial processing of Hex polypeptides were comparable in normal and affected fibroblasts, but mature polypeptides were quantitatively localized in "buoyant" lysosomes of affected cells, along with Hex activity; moreover, mature alpha-chain of Hex was approximately 2 kDa larger than that observed in normal cells. The molecular weight difference was apparently due to impaired proteolytic processing of alpha chain in affected fibroblasts, since treatment of immunoprecipitated alpha-chain from normal and affected cells with neuraminidase and endo-beta-N acetylglucosaminidase H failed to resolve the molecular weight difference. The impaired processing was observed to be persistent (after a chase of up to 200 h), but had no apparent effect on the turnover or activity of Hex in affected fibroblasts. The observed proteolytic processing defect may be primary or secondary in infantile NeuAc storage disease. PMID- 2965874 TI - IGF receptors in myocardial capillary endothelium: potential regulation of IGF-I transport to cardiac muscle. AB - Beating rat hearts were perfused with 125I-IGF-II alone or 125I-IGF-II and unlabeled IGF-II or insulin, then prepared for radioautography. Maximal 125I-IGF II grain counts over capillaries were decreased in a dose-dependent manner by unlabeled IGF-II but were unaffected by coperfusion with insulin. To determine a potential role for capillary receptors in the transfer of circulating IGF to cardiac muscle, the effects of sequential loss of capillary IGF binding sites was determined. For IGF-I, loss of capillary binding sites by trypsin perfusion was accompanied by proportional decreases in the subsequent appearance of IGF-I in cardiac muscle. In contrast, similar decrements of capillary IGF-II binding did not affect muscle levels of IGF-II. We conclude that capillary endothelium of the intact heart possesses distinct IGF-I and IGF-II binding sites, with the capillary IGF-I binding sites being of potential importance in the transfer of vascular IGF-I to subendothelial cardiac muscle. PMID- 2965875 TI - Methotrexate in rheumatoid arthritis. PMID- 2965876 TI - Ten-year follow-up study of patients with Yersinia arthritis. AB - Eighty-five patients with acute Yersinia arthritis were seen in followup for a mean of 10 years. During that time, peripheral joint symptoms occurred frequently (51.8%), but these symptoms were mild (45.9%). Development of a new reactive arthritis (4.7%) or chronic arthritis (2.4%) was uncommon. One-third of the patients experienced low back pain, and one-third of the patients had radiologic evidence of sacroiliitis. The presence of sacroiliitis was more frequent in patients with low back pain (46.7%) than in those who did not have symptoms (21.2%). More patients with HLA-B27 had low back pain and sacroiliitis, but there was no association of this genetic factor with the residual symptoms in peripheral joints. PMID- 2965877 TI - Increase in levels of serum keratan sulfate following cartilage proteoglycan degradation in the rabbit knee joint. AB - Following injection of chymopapain into a single knee joint in rabbits, serum keratan sulfate levels rose sharply and remained elevated for at least 48 hours before returning to preinjection levels. These changes were accompanied by depletion of proteoglycans from articular cartilage in the injected joint. We conclude that serum keratan sulfate levels rise predictably following acute loss of proteoglycan from a single joint. PMID- 2965878 TI - [The Chilean health system: organization, operation and financing]. PMID- 2965879 TI - [Financing of health policies in Brazil]. PMID- 2965880 TI - [Efficacy and efficiency of social security in relation to the economic cycle: the Peruvian case]. PMID- 2965881 TI - [Fees charged to health services users. Analysis of their potential effects in Peru]. PMID- 2965882 TI - [Promotion of breast feeding through an educational campaign directed at pregnant women]. PMID- 2965883 TI - [Epidemiologic survey of histoplasmosis, paracoccidioidomycosis and leishmaniasis using skin tests]. PMID- 2965884 TI - [Enterotoxigenic Escherichia coli and Campylobacter jejuni in acute diarrheic syndrome in Chilean infants]. PMID- 2965885 TI - [Heterosexual transmission of AIDS]. PMID- 2965886 TI - [Experimental model in rabbits for the correction of defects in the abdominal wall, using an autologous graft of the fascia lata]. PMID- 2965887 TI - [Alagille syndrome. Apropos of a case]. PMID- 2965888 TI - A compartment of effector helper and suppressor T cells in the normal mouse thymus. AB - We have recently described an autonomously activated set of T cells in the spleens of normal and "antigen-free" mice which display effector T helper (TH) or T suppressor (TS) activities; we describe here an intrathymic effector T-cell compartment which directly helps or suppresses B-cell responses and appears to be distinct from the peripheral set of effector cells. Splenic effector T cells do not represent recent thymic migrants (because adult thymectomized mice have unaltered levels of effector TH and TS cells in the spleen), nor do intrathymic effector T cells represent circulating peripheral T cells (since thymic effector T cells are B2A2+, while splenic effector T cells are B2A2-). Furthermore, effector TH cells within the two compartments exert differential effector activities: splenic effector TH cells induce B cells to both proliferation and maturation, while thymic effector TH cells are defective in activating B-cell maturation. The present findings extend our studies on "natural" lymphocyte activities in the normal immune system, revealing the existence of two apparently distinct effector T-cell compartments. The potential significance of the intrathymic set of effector cells in repertoire selection is considered. PMID- 2965889 TI - [Purification of phosphoglucomutase isoenzymes from human placenta]. PMID- 2965890 TI - Social gaze, social avoidance, and repetitive behavior in fragile X males: a controlled study. AB - Preference for social gaze as well as the percentage occurrence of social gaze, nonverbal social avoidance, and nonverbal repetitive behaviors were examined in autistic and nonautistic prepubertal males with the fragile X syndrome (fra[X]) during social interaction with a parent or stranger. Comparison groups were nonhandicapped, Down syndrome, atypical pervasive developmental disorder, and autistic males. The subjects with fra(X) and the nonhandicapped and Down syndrome control subjects discriminated parent from stranger as evidenced by their avoidance behavior. The overall percentage of avoidance was higher, however, for both parent and stranger, among the males with fra(X). Autistic and atypical groups without fra(X) failed to discriminate parent from stranger in their avoidance behavior. Possible explanations for these group differences in terms of language level or degree of language demand were ruled out. Implications for research concerning the relations among fra(X), autism, and mental retardation were discussed. PMID- 2965891 TI - Effects of age on the adaptive behavior of institutionalized and noninstitutionalized individuals with Down syndrome. AB - The effects of age on the adaptive behavior of individuals with Down syndrome and matched individuals in other diagnostic categories were compared in three studies. The only study that provided evidence of a decline in competence among older individuals with Down syndrome was the one study in which the oldest age category was 60 years and over. In that study, a significant Age X Diagnosis interaction was found for the Motor Development factor of the Client Development Evaluation Report, reflecting a sharp drop in the scores of some of the oldest subjects with Down syndrome. These results resemble those of other recent research in two respects: the age at which the decline in competence occurred and the nature of that decline. PMID- 2965893 TI - [Morphometric investigation on the neostriatum and spinal cord in extrapyramidal degenerative diseases and amyotrophic lateral sclerosis]. PMID- 2965892 TI - Deficiency of the autologous mixed lymphocyte reactions of non-T/T and T/T type in intravenous drug abusers infected by the human immunodeficiency virus (HIV). AB - In the present study both responsiveness and stimulatory capacity in autologous mixed lymphocyte reactions (AMLRs) of non-T/T and T/T type, as well as in allogeneic mixed lymphocyte reaction (MLR), were evaluated in 30 intravenous drug abusers (IDAs) infected by the human immunodeficiency virus (HIV) and in 10 HIV negative IDAs. The production of interleukin 2 (IL2), and the expression of HLA Class II antigens and IL2 receptors by PHA-activated T lymphocytes were also evaluated. A severe impairment of both responsiveness and stimulatory capacity in MLR and AMLRs was found in the HIV-positive IDAs and not in the HIV-negative IDAs. The HIV-positive IDAs showed also a defective expression of HLA Class II antigens, whereas the IL2 production and the IL2 receptor expression were in the normal range. The present data are consistent with similar observations in male homosexuals with AIDS-related complex and confirm that the HIV infection induces a broad spectrum of immunological abnormalities leading to a progressive derangement of the immunocompetence. PMID- 2965894 TI - The dental health of the elderly mentally ill: the carers' perspective. PMID- 2965896 TI - Ultrasonography of the kidney following renal biopsy in children. AB - The frequency of renal and perirenal hematomas following percutaneous renal biopsy using a cutting needle was studied by means of ultrasonography in 30 children and young adults aged 3-16 years. Hematomas occurred in 64.5 percent of the biopsies but caused significant clinical symptoms in one patient only. PMID- 2965895 TI - Current trends in the management of urinary stones. AB - The treatment of renal and ureteral stones has undergone rapid and major changes over the past ten years. Extracorporeal shockwave lithotripsy has become the most commonly used modality for the treatment of renal and upper ureteral stones. Lower ureteral stones are more commonly being approached by retrograde techniques. Percutaneous nephrolithotomy, medical therapy, and open surgical nephrolithotomy offer viable alternatives in given situations. Presented here is the current application of each of these techniques, both alone and in combination, for the treatment of urinary stones. PMID- 2965897 TI - Urographic renal size in acute pyelonephritis in childhood. AB - Eighteen children with acute pyelonephritis were investigated by urography at the time of and about two months after the acute infection. Renal size was evaluated as renal length and parenchymal area. During the acute infection renal size was larger than at the follow-up examination two months later. In conclusion, urography performed in close connection with the acute pyelonephritic infection gives an overestimation of renal size. PMID- 2965898 TI - Suprapubic micturition cystourethrography. AB - Suprapubic micturition cystourethrography is performed by puncturing the bladder aseptically, in the midline, 1 to 2 cm above the symphysis pubis with an 18 to 21 gauge needle. The needle is pointed 10 to 15 degrees cranially to avoid puncture of the trigonum and urethra. Anaesthesia, antibiotic protection, and diuretics are not necessary. In 225 examinations performed on 200 patients, the only complications noted were transient haematuria (1.5%) and slight extravasation of contrast medium through the puncture sites (8%). None required any therapeutic measures. In 4 patients puncturing of the bladder needed ultrasound guidance. The technique is ideal for the study of vesicoureteral reflux, neurogenic bladder, urethral strictures, posterior urethral valves and other urethral pathology. It is also useful in balloon catheter dilatation of urethral strictures. Suprapubic micturition cystourethrography is safe, efficient and time-saving. It is well accepted by the patients and carries little risk of urinary infection. PMID- 2965899 TI - Non-invasive measurement of stroke volume and left ventricular ejection fraction. Radionuclide cardiography compared with left ventricular cardioangiography. AB - The stroke volume (SV) was determined by first passage radionuclide cardiography and the left ventricular ejection fraction (LVEF) by multigated radionuclide cardiography in 20 patients with ischemic heart disease. The results were evaluated against those obtained by the invasive dye dilution or thermodilution and left ventricular cardioangiographic techniques. In a paired comparison the mean difference between the invasive and radionuclide SV was -1 ml (SED 3.1) with a correlation coefficient of 0.83 (p less than 0.01). Radionuclide LVEF values also correlated well with cardioangiographic measurements, r = 0.93 (p less than 0.001). LVEF determined by multigated radionuclide cardiography was, however, significantly lower than when measured by cardioangiography, the mean difference being 6 per cent (p less than 0.001). These findings suggest that radionuclide determinations of SV and LVEF are reliable. The discrepancy between the non invasive and invasive LVEF values raises the question, whether LVEF is overestimated by cardioangiography or underestimated by radionuclide cardiography. PMID- 2965900 TI - Computed tomography after lumbar disc surgery. AB - Computed tomography (CT) findings following lumbar disc surgery were analyzed in a series of 53 patients one week after surgery and in another series of 43 patients 6 months to 20 years (average 4.3 years) after surgery. In addition to changes caused by the surgical exposition (laminotomy or laminectomy, gas bubbles) a mass lesion was found on the posterior aspect of the disc in all cases. Hypodensity of the disc interspace as well as vacuolization were found in several cases. These changes were less in the patients examined late after surgery but the CT findings were never completely normal. Findings characteristic of recurrent prolapse included disc-like attenuation values within the mass and non-enhancement with contrast medium. Hypodensity of the disc interspace is associated with discitis but as a non-specific finding it is insufficient in itself for diagnosis of discitis. PMID- 2965901 TI - Juxta-articular erosions in reflex sympathetic dystrophy. AB - Thirty-one patients with documented reflex sympathetic dystrophy syndrome (RSDS) were reviewed for their radiographic changes. Juxta-articular and metaphyseal bone loss was found in the majority of the patients. Juxta-articular bone loss closely resembling erosions seen in rheumatoid arthritis was found in all the patients. The significance of these findings is discussed. PMID- 2965902 TI - Trochanteric fractures. Classification and mechanical stability in McLaughlin, Ender and Richard osteosynthesis. AB - Four hundred and thirty trochanteric fractures operated upon with McLaughlin, Ender or Richard's osteosynthesis were divided into 6 different types based on their radiographic appearance before and immediately after reposition with special reference to the medial cortical support. A significant correlation was found between the fracture type and subsequent mechanical complications where types 1 and 2 gave less, and types 4 and 5 more complications. A comparison of the various osteosyntheses showed that Richard's had significantly fewer complications than either the Ender or McLaughlin types. For Richard's osteosynthesis alone no correlation to fracture type could be made because of the small number of complications in this group. PMID- 2965903 TI - Ultrasonography and gallbladder perforation in acute cholecystitis. AB - A group of 24 patients with perforated gallbladder operated upon between 1980 and 1987 was compared with a group of 21 patients operated upon in 1982 due to uncomplicated acute cholecystitis in order to find out whether it was possible to use some specific sonographic signs to find the patients at risk of perforation. Free fluid and fluid collections close to the gallbladder fossa were found in 9 patients. The patients with perforated gallbladders tended to have a slightly thicker gallbladder wall--7 mm (range 3-20 mm)--when compared with the uncomplicated cases of acute cholecystitis--5.3 mm (range 2-13 mm). Localized fluid collection in the wall of a gallbladder was seen in a patient just prior to the perforation. It was, however, not possible to find a common sign characteristic for imminent perforation. The study showed, however, that the combination of early diagnosis with ultrasound together with aggressive surgery reduced the mortality at gallbladder perforation to none in the last 9-year period compared with 10 per cent in the previous 9-year-period. PMID- 2965904 TI - Fatty meal provocation monitored by ultrasonography. A method to diagnose ambiguous gallbladder disease. AB - Impaired gallbladder emptying is a recognized sign of acalculous gallbladder disease (AGBD). The emptying function was studied by ultrasonographic monitoring before and after ingestion of fat-containing food in a prospective study comprising 124 patients (136 examinations). In 17 patients the antero-posterior diameter of the gallbladder was unchanged or increased postprandially in addition to the onset of abdominal pain and nausea, signs interpreted as consistent with AGBD. Cholecystectomy, performed in 12 of these individuals, revealed histopathologic changes of different degree in 11 and adhesions of the gallbladder to adjacent organs in 8. The method is simple to perform, cheap, has no side effects and gives a low rate of false positive diagnoses. PMID- 2965905 TI - Catheter perforation of distal oesophagus with duodenal re-entry of catheter. Report of a case. AB - An unusual case of perforation of the oesophagus is presented. A nasogastric tube had perforated the oesophagus and re-penetrated into the duodenum, and thereby re entered the gastrointestinal tract without perforating the peritoneum and without causing the classical clinical signs of oesophageal perforation. Treatment was started 31 days after the perforation. PMID- 2965906 TI - Ultrasonically guided fine needle aspiration of suggested enlarged parathyroid glands. AB - Ultrasonically guided fine needle aspiration from 22 ultrasonically suspect enlarged parathyroid glands was performed in 21 consecutive patients with biochemically confirmed hyperparathyroidism. Histologic examination revealed parathyroid tissue in 20 and thyroid tissue in 2 of the 22 ultrasonically suspect parathyroid glands. The aspirated material (21 patients) was analysed for PTH content and compared with the PTH content in aspirates from corresponding thyroid tissue. In all but one, a higher PTH content was found in aspirates from parathyroid glands. Further, the aspirated material (16 patients) was cytologically examined for parathyroid cells. In 10 of 14 histologically confirmed parathyroid glands cells of parathyroid origin were found, but in 4 cases only cells of endocrine origin were seen. The results and the use of fine needle aspiration in relation to the ultrasonic findings are discussed. PMID- 2965907 TI - Effect of alcohol intake on the radiographic quality in patients with midfacial trauma. AB - The initial reports on radiologic examinations in 618 consecutive patients with midfacial injuries were compared with the final clinical diagnoses. Alcohol had to some degree been imbibed by 31 per cent of the patients. Influence of alcohol was 3 times more common outside than within office hours. Radiographic analysis included estimation of blurring, errors in straightness, angulation, beam centering and limitation. Image quality was scored as visibility of 'the imaginary lines of bony continuity'. There was no statistically significant correlation between the degree of inebriety and image quality or diagnostic performance with the radiographic technique used, with the patient supine. There is no need to postpone midfacial radiography in inebriate patients. PMID- 2965908 TI - Hyperostosis cranii. Radiography and scintigraphy compared. AB - The bone scintigrams of 52 women with radiographically proven hyperostosis cranii were examined specifically with regard to the endosteal lesions. It was confirmed that hyperostosis cranii exhibits a characteristic appearance at bone scintigraphy that is usually easy to recognize. By scoring the degree of hyperostotic changes and their scintigraphic activity according to a semiquantitative scale, an inverse relation between radiographic changes and scintigraphic activity was observed but could not be statistically proven. A tendency of more pronounced radiographic changes and a tendency of decreasing activity at bone scintigraphy with increasing age were also found. It is suggested that hyperostosis cranii starts in younger women and that the initial osseous turnover of the lesions is high. Successively, the metabolic activity decreases while the endosteal lesions increase, tending towards a stabilization of the process with increasing age. PMID- 2965909 TI - Magnetic resonance imaging and the acquired immunodeficiency syndrome dementia complex. AB - It is now recognized that patients infected by the virus linked to the acquired immunodeficiency syndrome (AIDS) can develop dementia symptoms as the initial and sometimes only symptomatology for AIDS. This appears to be a syndrome whose origin is independent of secondary non-viral infection or malignancy. Magnetic resonance imaging (MRI) in three patients with well documented AIDS dementia revealed high signal periventricular white matter lesions. In one case, large lesions were not apparent on computed tomography and gross inspection of the fixed brain prior to autopsy. In two cases in which serial in vivo MR studies were obtained, there was a progressive increase in lesion volume over a short (several months) period of observation. Periventricular white matter lesions may be an early sign accompanying AIDS dementia, and the degree of changes correlated well with the clinical picture in our patients. PMID- 2965910 TI - Comparison of different pulse sequences for in vivo determination of T1 relaxation times in the human brain. AB - Quantitative in vivo determination of T1 relaxation times by magnetic resonance imaging (MRI) is hampered by several potential sources of error. This study focused on the influence of the radiofrequency pulse sequences applied with special attention to the significance of the repetition time (TR). T1 measurements were performed on the human brain using a whole body MR scanner operating at 1.5 tesla. Three different pulse sequences were compared including two 6-points inversion recovery (IR) sequences with TR = 2.0 s and 4.0, respectively, and a 12-points partial saturation inversion recovery (PSIR) sequence with TR varying between 0.24 and 8.0 s. The median T1 relaxation times obtained in cortical grey matter and cerebrospinal fluid were significantly shorter in the IR experiments at TR = 2 s than in those carried out at TR = 4 s. Concerning white matter the discrepancy was much less pronounced, but still statistically significant. Supplementary phantom measurements indicated that the higher T1 values are increasingly underestimated when TR is reduced to 2 s. The results suggest that the PSIR sequence or IR sequences with a TR greater than 2 X the T1 level of the tissue type investigated should be employed for accurate T1 determination by MRI in clinical work. PMID- 2965911 TI - Local blood-brain barrier penetration following systemic contrast medium administration. A case report and an experimental study. AB - The present study was initiated by a severe complication in a patient with renal dysfunction who developed cortical blindness and weakness of her left extremities 30 hours following renal and abdominal angiography. To evaluate the impact of prolonged high serum concentrations of contrast medium (CM) this clinical situation was simulated in a laboratory model using sheep with elevated serum levels of contrast medium maintained for 48 hours. The experimental data did not support the theory that the prolonged exposure to high circulating levels of contrast medium (4 ml/kg body weight of meglumine diatrizoate 60%) is sufficient alone to cause penetration of the blood-brain barrier. PMID- 2965912 TI - Ionic and non-ionic contrast media used for contrast-enhanced computed tomography in experimental pancreatitis. AB - Contrast enhancement of the pancreas was studied in pigs using dynamic computed tomography in experimental oedematous and haemorrhagic/necrotizing pancreatitis during the first two minutes after injection of an intravenous bolus of non-ionic contrast medium (iohexol). The prospects of separating the two forms of the disease, known to be possible with ionic contrast media, were tested with a non ionic contrast medium. In the oedematous form, contrast enhancement after 5 hours of the disease was significantly higher than in the haemorrhagic/necrotizing form. Contrast enhancement after 30 hours of disease tended to vary with the severity of the disease, showing that the course of oedematous pancreatitis is dynamic. Intermediate forms occur and follow-up studies are needed during the disease. A non-ionic contrast medium proved as good for separating the two forms of the disease in the early phase as were ionic contrast media. In severely ill patients, non-ionic contrast media should therefore be used. PMID- 2965913 TI - Addition of local anesthetics to contrast media. II. Increase of acute mortality in mice with intravenous contrast administration. AB - The acute intravenous toxicity (i.v. LD50) of solutions of the ratio 1.5 contrast media metrizoate or diatrizoate and the ratio 3.0 contrast medium metrizamide was determined in mice with and without the addition of local anesthetics to the solutions. The two local anesthetics mepivacaine or lidocaine were added to final concentrations up to 2.0 mg/ml of the contrast medium solutions. This corresponds to clinically used concentrations. All additions of local anesthetics to the solutions increased the mortalities caused by the contrast medium solutions. Addition of local anesthetics to a final concentration of 2 mg/ml approximately doubled the acute intravenous toxicity of the contrast media. The ratio 3 contrast media produce less hypertonic solutions than the ratio 1.5 contrast media and should be preferred for angiography because they cause less pain and do not require the addition of local anesthetics which increase the acute toxicity of the solutions. PMID- 2965914 TI - Bone scintigraphy in staging of bladder carcinoma. AB - The role of bone scintigraphy in the staging of bladder carcinoma before attempted radical therapy was evaluated from a personal series of 71 consecutive cases. The results from this were correlated to previous reports--accumulated data from a total of 458 staging bone scans were thus obtained. In this accumulated material metastases were diagnosed in 4.6 per cent--in 2.8 per cent true positive and in 1.7 per cent false positive findings. The effect of the bone scanning results on therapy was minimal: cystectomy was performed in spite of the diagnosis of metastases in 16 out of 21 patients. In only 4 patients (0.9%) surgery was avoided because of scintigraphy results. Scintigraphy thus has no place in the routine preoperative staging of bladder carcinoma. PMID- 2965915 TI - Assisting with infectious patients. PMID- 2965916 TI - Ferritin-bearing T-lymphocytes and serum ferritin in patients with breast cancer. AB - Flow cytometric studies of T-lymphocytes in breast cancer patients show that the number of cells bearing ferritin on their surface is significantly greater than normal. The number of ferritin-bearing T-cells does not appear to be related to the clinical stage of the disease nor to the serum ferritin concentration, though this is higher in cancer patients than in normal women. There is no difference in the number of T-cells positive for interleukin 2 or transferrin receptors nor in the absolute number of T-cells, T-helper cells and B-cells between normal women and those with breast cancer or benign breast disease. However, there is a significant increase in the level of HLA DR-positive T-cells and T-suppressor cells in breast cancer patients. While the significance of ferritin-bearing T cells is not known an increase in their number appears to be associated with cancer. PMID- 2965917 TI - Utilization of tryptophan, nicotinamide and nicotinic acid as precursors for nicotinamide nucleotide synthesis in isolated rat liver cells. AB - 1. Incubation of isolated rat hepatocytes with nicotinamide or nicotinic acid showed that while both vitamers were taken up from the incubation medium, neither was utilized to any significant extent as a precursor of the nicotinamide nucleotide coenzymes, NAD and NADP, and neither was capable of preventing the loss of nucleotides that occurs on incubating the cells. 2. Incubation of hepatocytes with tryptophan showed that de novo synthesis from tryptophan permitted replacement of the nucleotides lost during incubation; at high concentrations of tryptophan there was an increase above the initial intracellular concentration of NAD(P). Incubation of hepatocytes with tryptophan also resulted in the formation and release from the cells of a considerable amount of niacin, as well as the two principal metabolities of NAD(P), N1-methyl nicotinamide and methyl pyridone carboxamide. 3. It is suggested that, in the liver, performed niacin is not utilized for nucleotide synthesis, and indeed the function of the liver appears to be synthesis of niacin from tryptophan, and its release for use by extrahepatic tissues that lack the pathway for de novo synthesis of nicotinamide nucleotides from tryptophan. PMID- 2965918 TI - [Increased substrate selectivity during transition from Ca2+-activated to K+,EDTA activated nucleoside triphosphatase activity of heavy meromyosin]. AB - A comparison of kinetic parameters (Km(app) and V) of hydrolysis by heavy meromyosin of natural (ATP and ITP) and modified nucleoside triphosphates showed that in the K+, EDTA-ATPase conformation the enzyme exhibited a higher selectivity towards the structure of the substrate nucleoside moiety than in the case of the Ca2+-stimulated nucleoside triphosphatase activity. In the presence of Ca2+, all the N1- and N6-substituted analogs of ATP as well as ITP, etheno-ATP and the dialdehyde derivative of ATP were hydrolyzed at a high rate irrespective of their markedly decreased affinity for heavy meromyosin. In the presence of K+, EDTA the ATPase activity showed a tendency for a total decrease of the analog affinity for nucleoside triphosphates, i.e., the impossibility of tight binding of the substrate phosphate residues to the protein in the absence of bivalent cations, which was concomitant with an increase in the hydrolysis rate. However, it was found that only in N1-substituted analogs any appreciable changes in the substrate properties were absent. All the other nucleoside triphosphates tested (N6-carboxy-methoxy-ATP, N6-(N'-acetylaminoethoxy)-ATP, etheno-ATP, ITP and the dialdehyde derivative of ATP having a rupture in the ribose ring) lost their ability to be hydrolyzed by heavy meromyosin. The experimental results as well as the literature data are suggestive of differences in the spatial structure of the active center in two different myosin conformations associated with a high catalytic activity, i.e., K+, EDTA-ATPase and Ca2+-ATPase. PMID- 2965919 TI - [Regulation by alcohols and anions of soluble ATPase activity in chloroplasts and mitochondria]. AB - The alcohols and anions present at the medium before the beginning of ATPase reaction increased or decreased the lag-time of ATPase hydrolysis by chloroplast coupled factor according to the extent of their inhibitory or stimulating action. The effect of alcohols and anions on the ATPase activity of coupled factor of chloroplasts and mitochondria develops less than 2 s in case of their introduction during the ATP hydrolysis. PMID- 2965920 TI - [Changes in the population structure of the red-tailed Libyan jird as affected by toxicants]. AB - The kind of the regulating agent for the quantitative control of the epidemically dangerous species depends on the concrete tasks. The poison of the critical action causes a strong, but short-time effect. The pesticide inhibiting the reproductive system of the rodents exerts long and profound influence on the population structure. PMID- 2965921 TI - Two-dimensional NMR studies of [Pro-10] atrial natriuretic factor [7-23]. PMID- 2965922 TI - [Controlling effect of T lymphocyte suppressors on the proliferation of cells in various tissues]. AB - The changes in mitotic index of mouse bone marrow, spleen, liver, kidneys, small intestine, cornea were studied during alterations in the number of T-suppressors. It was found that mitotic activity in all tissues investigated enhanced significantly after a decrease in the number of T-suppressors caused by single injection of an antiserum against T-suppressors. On the contrary, the mitotic index diminished significantly after the transfer of tolerant spleen lymphoid cell suspension enriched by T-suppressors to normal syngeneic mice. These data indicate that T-suppressors are responsible for the control over cell proliferation in nonlymphoid tissues. PMID- 2965923 TI - [Blockade of adrenoreceptors--a mechanism for impairing the humoral and cellular regulation of the proliferation of epidermal cells in psoriasis]. PMID- 2965924 TI - Imbalances within the peripheral blood T-helper (CD4+) and T-suppressor (CD8+) cell populations in the reconstitution phase after human bone marrow transplantation. AB - Peripheral blood T cell subsets were evaluated in 11 patients during the reconstitution phase after allogeneic bone marrow transplantation and compared with 11 age-matched controls. The proportion of cells coexpressing Leu7 and CD11b (C3bi receptor) markers was determined within the CD4+ (T-helper) and the CD8+ (T suppressor) subsets by two-color immunofluorescence analysis. CD4+ and CD8+ T cells reached normal or near-normal values within the first year posttransplant. In contrast to normal controls, however, most of the cells in both subsets coexpressed the Leu7 and CD11b markers. T cells with such phenotype display the morphological features of granular lymphocytes (GLs) and a functional inability to produce interleukin 2 (IL 2). These T cell imbalances were not related to graft v host disease (GvHD) or to clinically detectable virus infections and may account for some defects of cellular and humoral immunity that occur after bone marrow transplantation. PMID- 2965925 TI - Behavior of plasminogen at the luminal surface of the normal and deendothelialized rabbit aorta in vivo and in vitro. AB - The behavior of purified rabbit plasminogen at the luminal surface of the uninjured and deendothelialized rabbit aorta has been studied in vivo and in vitro. After intravenous injection, 125I-plasminogen associated rapidly with the endothelium (approximately 0.1 pmol/cm2 at saturation) and passed through to accumulate in the subendothelium. At two to 15 hours after injection, 11 to 15 times more radioactivity was associated with the subendothelium than with the endothelium. Removal of the endothelium by balloon catheter led to a rapid adsorption of 125I-plasminogen by the luminal surface of the vessel; saturation (9.1 pmol/cm2) was attained at ten to 20 minutes after deendothelialization. Of the adsorbed plasminogen (radioactivity), only 2% to 4% was associated with the adherent platelet monolayer. Uptake of 125I-plasminogen by the deendothelialized vessel was not significantly inhibited by epsilon-aminohexanoic acid whether injected before or after the 125I-plasminogen. No evidence of plasmin activity at the aorta surface was found from either transmission electron microscopy studies or from amidolytic assays of plasminogen-saturated deendothelialized aorta samples before or after urokinase treatment. Balloon catheter treatment in vivo, however, generated significant antiplasmin activity of the deendothelialized aorta surface. We conclude that plasmin formed in vivo is probably inactivated by the antiplasmin activity that is associated with the subendothelium. PMID- 2965926 TI - Clonal rearrangement of the T cell receptor beta gene in the circulating lymphocytes of erythrodermic follicular mucinosis. AB - Follicular mucinosis is a condition characterized by the abnormal accumulation of acidic mucopolysaccharides in hair follicles. It is classically described as occurring idiopathically in young persons and within the infiltrates of mycosis fungoides in older individuals. We report a 12-year-old girl who had erythrodermic follicular mucinosis, hypereosinophilia, circulating Sezary cells, and both immunophenotypic and genotypic evidence of T cell neoplasia. Erythrodermic follicular mucinosis may represent an unusual variant of the Sezary syndrome, which to date has not been described in children or adolescents. PMID- 2965927 TI - Increased resistance to plasmic degradation of fibrin with highly crosslinked alpha-polymer chains formed at high factor XIII concentrations. AB - We have previously demonstrated that increasing factor XIII concentrations above that present in plasma (1 U/mL) results in the formation of very high molecular weight alpha fate polyacrylamide and agarose gel electrophoresis (SDS-PAGE). In this report, we have examined the effect of such crosslinking on plasmic susceptibility of fibrin prepared from purified fibrinogen and from plasma in the presence of factor XIII concentrations between 0 and 10 U/mL. The crosslinking achieved with purified fibrinogen at 1 U/mL factor XIII increased resistance to plasmic degradation by 32% as measured in a radiolabeled clot lysis system. However, further increases in plasmic resistance occurred at factor XIII concentrations of 2 and 10 U/mL, the latter decreasing the lysis rate to 45% of that which occurred in the absence of factor XIII. To achieve the same rate of clot lysis with fibrin formed using 10 U/mL rather than 1 U/mL of factor XIII, an increase in plasmin concentration of up to 4.2-fold was required. Similar results were obtained using clots prepared from plasma in the presence of factor XIII concentrations greater than 1 U/mL. Since the alpha 2-plasmin inhibitor content was the same for fibrin at 1 or 10 U/mL factor XIII, the increasing plasmic resistance could not be attributed to increased binding of the inhibitor. We conclude that fibrin prepared in the presence of factor XIII at concentrations exceeding that in plasma shows increased resistance to plasmic degradation, which is likely explained by the formation of very high molecular weight alpha polymer chains. PMID- 2965928 TI - Hemostatic enzyme generation in the blood of patients with hereditary protein C deficiency. AB - The presence of hereditary protein C deficiency has been shown to predispose patients to the development of venous thrombosis. We used radioimmunoassays for the protein C activation peptide (PCP) and the prothrombin fragment F1 + 2 to quantitate the extent of in vivo activation of protein C by thrombin thrombomodulin and prothrombin by factor Xa, respectively, in the blood of individuals with this clinical disorder. A total of 46 protein C deficient subjects from 18 kindreds were studied. In 23 nonanticoagulated patients with an isolated deficiency of protein C, the mean level of PCP was substantially reduced while the mean concentration of F1 + 2 was significantly elevated as compared with normal controls (1.10 pmol/L v 1.78 pmol/L, P less than .0005 and 2.54 nmol/L v 1.51 nmol/L, P less than .0005, respectively). The metabolic behavior of 131I-F1 + 2 was found to be similar in protein C deficient patients and normal individuals. However, we were unable to establish a significant correlation between decreased PCP levels and increased F1 + 2 measurements in these 23 patients. This study demonstrates that heterozygous protein C deficient individuals with equivalent plasma levels of the zymogen may have markedly different biochemical profiles when assay techniques are used that quantitate the in vivo activity of the coagulation system. Six individuals from three pedigrees were identified as having combined deficiencies of protein C and either antithrombin III or protein S; the genetic basis for the combined deficiency state was determined in two of the kindreds. Finally we observed that hemostatic system activity as measured by the PCP and F1 + 2 assays is markedly suppressed in protein C deficient patients who are chronically anticoagulated with coumarin derivatives. PMID- 2965930 TI - High numbers of CD4+ T cells showing abnormal recognition of DR antigens in lymphoid organs involved by Hodgkin's disease. AB - Purified T lymphocytes (E rosetting cells) isolated from the involved lymphoid organs (lymph nodes and spleen) of five patients with Hodgkin's disease (HD) were cloned under culture conditions (phytohemagglutinin plus interleukin-2) that allow clonal expansion of most T lymphocytes. A total number of 104 CD4+ T cell clones so obtained were tested for their ability to proliferate in response to autologous mitomycin-treated non-T cells. About half of these clones but none of 234 CD4+ T cell clones derived from normal lymphoid tissues or peripheral blood displayed a proliferative response to autologous stimulators. When clones proliferating in autologous mixed lymphocyte reaction (AMLR) were assessed for their ability to respond in allogeneic MLR (allo-MLR), most of them were found to exhibit consistent proliferation in response to more than one haplotype. Both the AMLR and the allo-MLR by HD clones were inhibited by adding monoclonal antibodies (MoAbs) reactive with monomorphic determinants of major histocompatibility complex (MHC) class II (DR) antigens to the cultures, whereas MoAbs reactive with MHC class I antigens were without effect. These studies suggest that lymphoid organs involved by HD contain high proportions of CD4 T cells showing abnormal recognition of DR antigens. These unusual cells may play an important role in the pathogenetic mechanisms occurring in HD. PMID- 2965931 TI - Fine structure of Blastocrithidia culicis as seen in thin sections and freeze fracture replicas. AB - The fine structure of epimastigotes of Blastocrithidia culicis was studied by transmission electron microscopy of thin sections and freeze-fracture replicas. This parasite presents a well developed endoplasmic reticulum and Golgi complex systems. Differences in the density and organization of the intramembranous particles were observed between the membranes which enclose the cell body and the flagellum. Ridge-like elevations, visualized in freeze-fracture replicas, were observed in sites where the mitochondrial branches touched the plasma membrane. A special array of membrane particles was observed on both faces of the flagellar and the cell body membranes at the region where the flagellum adheres to the cell body. It appeared as strands made of two rows of membrane particles. Filipin treated cells were used for the localization of membrane sterols in freeze fracture replicas. The number of filipin-sterol complexes varied from cell to cell. In some cells, rows of filipin-sterol complexes were seen. No complexes were observed in the region of the attachment of the flagellum to the cell body. PMID- 2965929 TI - Cellular and molecular studies on infant null acute lymphoblastic leukemia. AB - We have studied the cellular and molecular basis of eight cases of infant null acute lymphoblastic leukemia (ALL). All eight patients were under 9 months of age and presented with leukocyte counts in excess of 60 X 10(9)/L, organomegaly, and in two cases CNS infiltration. Although seven cases were morphologically classified as ALL, one patient had both lymphoid and myeloid features. Phenotypic analysis of leukemic blasts from all patients showed a typical null ALL pattern, ie, CD10 (common ALL antigen)-negative, strongly HLA-DR-positive, and CD19 (B4) positive. The presence of terminal deoxynucleotidyl transferase (TdT) at presentation was positive in six patients' cells and negative in two. Two patients also expressed the myeloid-associated markers CD33 (MY9) and CD15 (TG1), and coexpression of CD19 and CD33 was confirmed in these two by using dual marker flow cytometry (fluorescence-activated cell sorting). Electron microscopic examination of the same two patients' cells showed the presence of monocytoid blasts that labeled with the pan-B cell antibody B4 (CD19). Short-term culture of one of these patients cells in the presence of phorbol ester resulted in the majority of the cells exhibiting myeloid markers, strong nonspecific esterase positivity, and phagocytic properties. Cytogenetic analysis showed the common feature in 7 of 8 cases to be a break in band 11q23. Molecular analysis of DNA from the blast cells of all eight patients showed rearrangement of the immunoglobulin heavy-chain genes in all cases without, however, any evidence of kappa light-chain rearrangement. T cell receptor genes were present in the germline configuration in all cases. Rearrangements of the c-ets 1 oncogene, which maps to band 11q23, were not detected, thus providing no evidence for involvement of this oncogene in the common disease process. Our data indicate that although infant null ALL may present as a heterogeneous disease the similarity of many features between cases suggests a common derivation from a precursor cell sharing phenotypic and genotypic features of both B and myeloid progenitor cells. PMID- 2965932 TI - Stress response in Drosophila subobscura. III. Variability of heat shock puffs. AB - Variability in the heat shock response has been studied in D. subobscura, both in individuals from different strains at the same moment of development and in individuals belonging to the same strain at different moments of prepupal development. The heat shock response was independent of the genic background of the individuals, whereas a variability depending on the moment of development has been obtained. In relation to this variability two kinds of heat shock puffs have been found: those induced independently of the moment of development and those induced with a response dependent on it. In both cases a differential genic response that is dependent on the temperature of treatment has been detected. PMID- 2965933 TI - Distribution of nuclear matrix proteins in interphase CHO cells and rearrangements during the cell cycle. An ultrastructural study. AB - The nuclear matrix contains a group of residual non-histone proteins which remain structurally organized after extensive extraction of isolated nuclei with a high salt buffer, nucleases and a non-ionic detergent. Electron microscopic examination shows that the nuclear matrix is composed of a pore-complex lamina, an intranuclear network and residual nucleoli. In CHO cells biochemical analyses performed by one-dimensional SDS-PAGE show three major nuclear matrix polypeptides with molecular weights between 60 and 70 kDa. Polyclonal antibodies produced against these polypeptides were used to determine their nuclear distribution. Using immunoblotting, these proteins were found in whole nuclei, nuclear matrix, and in the intranuclear network but not in the pore-complex lamina. In order to determine the relationship between these structural proteins and the organization of the nucleus, the proteins were localized in situ. Ultrastructural detection was carried out by immunogold staining of thin sections of Lowicryl K4M-embedded cells. In interphase nuclei all condensed chromatin clumps were labelled. The nucleolus and the interchromatin granules were never immunogold-stained. During mitosis, the label was found to be associated with the chromosomes. This study shows that unlike the lamins, these 60-70 kDa nuclear matrix proteins are associated with the condensed chromatin throughout the cell cycle. PMID- 2965934 TI - Nuclear antigens differentially expressed during early development of Drosophila melanogaster. AB - To obtain specific immunological probes for studying molecular events involved in gene activation during early development of Drosophila, we have produced monoclonal antibodies directed against nuclear proteins differentially expressed in time and space during embryogenesis. Twenty-five antibodies against nuclear antigens detected after the onset of zygotic genome transcription were obtained. These antigens have been distributed into six categories depending on their reactivity in dot-blot, in Western-blot and in immunofluorescence on embryo frozen section assays. Six antigens (fifth and sixth categories) are stage-, but not tissue-specific and are present in all nuclei of the embryo after the blastoderm stage. Ten antigens, the third and fourth categories, did not react on immunofluorescence and have not been characterized for a possible tissue specificity. Antigens of the first and second categories are stage- and tissue- specific. Two of them are of particular interest. The first (250 kd, recognized by the monoclonal antibody GB7) is preferentially expressed in nuclei of the ventral nerve cord, whereas the other (65 kd, recognized by the monoclonal antibody LA9) is located in nuclei of the gut and associated structures. PMID- 2965935 TI - Effects of estramustine, a new anti-microtubule drug, on the induction of casein gene expression by prolactin. AB - Estramustine, a new anti-microtubule drug, was added to the culture medium of rabbit mammary explants with lactogenic hormones. In the absence of the drug, prolactin with insulin and cortisol stimulated DNA synthesis and it induced beta casein and beta-casein mRNA accumulation in the tissue. As opposed to other anti microtubule agents such as colchicine, estramustine was unable to prevent prolactin actions. An examination of the mammary cells by immunofluorescence revealed that the microtubule network was significantly altered under the influence of estramustine. These data indicate that the integrity of microtubules is not required for prolactin to deliver its message to the mammary cell. These data also suggest that other anti-microtubule drugs such as colchicine which prevent prolactin action act through their binding to tubulin molecule unrelated to microtubule structures. PMID- 2965937 TI - A mucus-secreting human colonic epithelial cell line responsive to cholinergic stimulation. AB - The human colonic epithelial cell line Cl.16E grows in culture as a polarized monolayer which differentiates at confluency into typical goblet cells secreting their mucin content into the culture medium. Polyclonal antibodies raised against these mucins were used in an ELISA to measure the amount of mucins secreted by the Cl.16E cells. Carbachol caused a transient and significant increase in mucus secretion with a maximal stimulation occurring at 30 min. A dose-dependent effect was found with a maximal stimulation with 10-3M carbachol. This effect was inhibited by atropine. These results indicate that the effects of carbachol are mediated by muscarinic receptors present on mucus-secreting epithelial cells. PMID- 2965936 TI - Effect of cetiedil on acetylcholine release and intramembrane particles in cholinergic synaptosomes. AB - The release of acetylcholine (ACh) from instantly frozen Torpedo electric organ synaptosomes in the course of stimulation is systematically associated with an increase in the number of large intramembrane particles counted on freeze fracture replicas. The drug cetiedil, which is a potent inhibitor of ACh release, also blocks the increase in the number of large particles. The blockage was studied either after ionophore A 23187 or Glycera neurotoxin action in the presence of calcium. PMID- 2965938 TI - Immunohistochemical localization of type IV collagen in mouse tooth germ: an ultrastructural study. AB - Localization of type IV collagen was analyzed at the ultrastructural level in mouse embryonic molars by using a preembedding technique. Cryostat sections were incubated with type IV collagen antibody and then treated with the peroxidase antiperoxidase complex. This antibody was visualized at the epithelio-mesenchymal interface. Labeling was intense and uniformly distributed throughout the basement membrane. However, it was mainly restricted to the lamina densa. No immunostaining was detectable in the lamina lucida but it was crossed by fine filaments that appeared as projections from the lamina densa to the epithelial cell plasma membrane. At the mesenchymal aspect of the basement membrane, projections of labeled material extended from the lamina densa in the underlying dental mesenchyme. At the presecretory stage of odontoblasts, these projections were in close connection with mesenchymal cell processes. PMID- 2965939 TI - The histologist Sigmund Freud and the biology of intracellular motility. AB - Sigmund Freud, the acclaimed founder of psychoanalysis, invested nine years of his early scientific effort investigating animal histology, cell biology and basic neuroscience prior to concentrating on human nervous and mental disorders. Through his histological studies Freud provided coherent evidence suggesting that the protoplasm consists of a contractile fibrillary network, the present-day cytoskeleton; he was one of the original founders of the fibrillary theory on the structure of the protoplasm. Concerning the biology of the cell nucleus, Freud appears to have been the first author who documented movements of nucleoli in nerve cells, a phenomenon presently referred to as nuclear rotation. In certain instances, Freud's observations antedate later views by more than half a century and are important to our current understanding of cell structure and basic processes of intracellular motility. PMID- 2965940 TI - The occurrence of metals Al, Fe, Ni, Cu, Zn in the nuclei of animal cells: an ultrastructural, in situ, X-ray microanalytical study. AB - Cell nuclei may contain significant quantities of the metals Fe, Ni, Cu, Zn, since they are present in the nucleo-enzymes and/or nucleic acids. These metals have been detected by X-ray microanalysis in situ in dinoflagellates (Kearns et al). Aluminum was only detected in cell nuclei in cases of natural or provoked intoxication. We observed at the ultrastructural level, in situ, the presence of Al, Fe, Ni, Cu, Zn in nuclei of different types of non-intoxicated animal cells. Moreover, we measured the concentration of these metals in the nucleolus and chromatin and compared it with the concentration of P and S. PMID- 2965941 TI - Characterization of a 30,000-35,000 m.w. monokine involved in the specific immune response: intracellular production, secretion and partial purification. AB - Peritoneal macrophages from normal mice which do not secrete interleukin 1 (IL 1) spontaneously release a factor with an approximate m.w. of 30,000-35,000. In contrast, in the presence of silica only IL 1 is produced. IL 1 as well as this macrophage-replacing factor (MRF) can restore the antibody response of macrophage depleted spleen cells. IL 1, however, is the only one that has the capacity to increase the proliferation of thymocytes. In every strain of mice studied, including nude mice, we observed a spontaneous release of MRF and a silica induced shift to the secretion of IL 1, except in lipopolysaccharide (LPS) hyporesponsive C3H/HeJ mice. Macrophages from these mice are unable to secrete MRF spontaneously, or IL 1 when stimulated with silica. The kinetics of the secretion of MRF and IL 1 appear to be similar. Macrophages, regardless of whether they have been stimulated to secrete IL 1, produce an intracellular IL 1 like activity with an approximate m.w. of 15,000. In contrast, the intracellular PFC-restoring activity is widely distributed in the 15,000-60,000 m.w. range; one of these compounds could be related to IL 1 precursor and/or to MRF itself. Chromatofocusing and chromatography on Blue Trisacryl have led to a partial purification and resolution from a possible contamination by IL 1. Purified MRF induces, in conjunction with lymphokines, the differentiation of B cells into antibody-forming cells. PMID- 2965942 TI - Precise localization of an overproduced periplasmic protein in Escherichia coli: use of double immuno-gold labelling. AB - The subcellular localization of beta-lactamase produced by a secretion-cloning vector pIN-III was studied by immunolabelling of frozen thin sections of Escherichia coli. Using double immuno-gold detections and internal reference proteins, it is shown here that beta-lactamase encoded by this vector can be exported and that its overproduction leads to aggregation within the periplasm. This aggregation induces the appearance of electron-dense areas immunolabelled by the antiserum directed against the beta-lactamase at the external side of the cytoplasmic membrane. The overproduced enzyme is also secreted to the medium in vesicles budding from the outer membrane of lpp strains. PMID- 2965943 TI - Dihydrocytochalasin B promotes adipose conversion of 3T3 cells. AB - Differentiation of preadipose 3T3-F442A cells into adipose cells is accelerated by the addition of dihydrocytochalasin B. The effect of the drug on 3T3-C2 cells is more marked: these cells are practically unable to differentiate in the absence of H2CB but a long-term exposure to the drug enables the cells to accumulate lipid droplets in medium supplemented with fetal calf serum and insulin. During their differentiation under these conditions the 3T3-C2 cells develop markers typical of adipose cells: glycerophosphate dehydrogenase, ATP citrate lyase, fatty acid synthetase and glycerophosphate acyltransferase. PMID- 2965944 TI - Calbindin D-27 kDa: preferential localization in non-B islet cells of the rat pancreas. AB - The presence and abundance of calbindin in rat pancreatic islet cells was assessed by immunohistochemistry of either whole islets or purified B and non-B islet cells, as well as by Western blotting of extracts derived from whole islets and purified B and non-B islet cells. Immunohistochemistry of pancreatic sections indicated a higher calbindin content in non-B cells, located at the periphery of the islets, than in the centrally located insulin-producing B cells. Comparable results were obtained in purified islet cells. Likewise, scanning densitometry of the Western blots indicated that, relative to cell volume, the single calbindin band (Mr 27 kDa) was 5-7 times higher in non-B than in B cells. In the splenic lobe of chick pancreas, however, the opposite situation prevailed. Thus, insulin producing cells clustered in small roundish islets were more intensely labelled after exposure to anti-calbindin serum than non-B islet cells located in large and irregularly shaped islets. Nevertheless, even in the chick pancreas, non-B islet cells contained an appreciable amount of calbindin. PMID- 2965945 TI - Effects of electrical stimulation upon post-hatching development of fibre types in normally innervated fast and slow latissimus dorsii muscles of the chicken. AB - In chicken, the main characteristic properties of muscle fibre types in slow anterior (ALD) and fast posterior (PLD) latissimus dorsii are acquired during post-hatching development. At day 4 it becomes possible to distinguish between alpha' and beta' fibre types in ALD muscle. At the same time, mATPase staining and NADH-TR activity permit recognition of alpha w and alpha R fibres within PLD muscle. During further development, muscle fibre typology progressively changes towards the adult slow and fast type. Chronic stimulation at a slow rhythm (5 Hz) of PLD prevents the change in relative proportions of alpha R and alpha W fibres within the muscle that occurs in normal post-hatching development and increases the number of beta R fibres. Moreover, oxidative activity is increased in all muscle fibre types following stimulation. In ALD muscle, chronic stimulation at a fast rhythm (40 Hz) results in a decrease in oxidative activity and inhibits the differentiation of alpha' and beta' muscle fibre types. This study demonstrates that in young chicken, the pattern of activity influences the differenciation of fibre types in slow and fast muscles. PMID- 2965946 TI - The energetic growth yields of the yeast Candida parapsilosis. AB - The energetic growth yields of Candida parapsilosis were compared with those of Saccharomyces cerevisiae as a function of the energy source in the presence or absence of antimycin A, an inhibitor of the second phosphorylation site. When glycerol was used as energy source, the energetic growth yields were quite similar in C. parapsilosis and S. cerevisiae. On the other hand, when experiments were carried out with glucose as energy source, although three phosphorylation sites were available, glucose was found to be a poor energy source for C. parapsilosis. When C. parapsilosis was grown in the presence of antimycin A, on glucose: YGluS = 3YGlu + AS and on glycerol: YGlyS = 2 YGly + AS. It was concluded that growth in the presence of antimycin A could occur due to the functioning of the third phosphorylation site. This result agrees with previous works indicating that in C. parapsilosis the alternative pathway merges into the main respiratory chain at the cytochrome c level. Although the doubling time of C. parapsilosis was much less temperature-sensitive than that of S. cerevisiae, the energetic growth yield was the same at 13 degrees C and 28 degrees C, and consequently, the secondary pathway did not seem to be thermogenic. PMID- 2965947 TI - AIDS update. PMID- 2965948 TI - Sexual hypersensitivity. AB - A variety of genital conditions appear to be precipitated by sexual intercourse in susceptible individuals. Some of these conditions represent true immunological hypersensitivity reactions whereas others have a nonimmunological basis and reflect an increased reactivity or sensitivity to sex by the individual concerned. This article introduces the term "sexual hypersensitivity" as a means of classifying these reactions. PMID- 2965949 TI - The current place of coronary angioplasty. AB - The number of coronary angioplasty procedures performed worldwide has risen exponentially during the last decade partially reflecting a wider scope for the technique. This article reviews the present indications for coronary angioplasty. PMID- 2965950 TI - Studies on the mechanism by which tryptophan efflux from isolated synaptosomes is stimulated by depolarization. AB - 1. The efflux and influx of tryptophan across the synaptosomal plasma membrane has been studied under a variety of experimental conditions, in order to examine the mechanism by which depolarization enhances the efflux of tryptophan from superfused synaptosomes. 2. Efflux of [3H]-tryptophan from preloaded superfused synaptosomes was found to be enhanced by K+ depolarization in a Ca2+ and dose dependent manner. In contrast, [3H]-phenylalanine efflux was only poorly stimulated by depolarization and only by very high concentrations of K+. 3. Tryptophan efflux was also enhanced by decreasing the extracellular Na+ concentration, but this effect was not dependent on extracellular Ca2+. 4. Influx of [3H]-tryptophan into synaptosomes was stimulated by extracellular Na+ removal, but the uptake of [3H]-phenylalanine was unaffected by this procedure. 5. Both the induced influx and efflux of tryptophan observed under these experimental conditions was inhibited by immobilizing the plasma membrane carrier with parachlorophenylalanine. This implied that both the enhanced influx and efflux arose as a consequence of the activation of the membrane tryptophan carrier, the direction of the observed effect being dependent upon the manner in which the experiments were conducted. 6. The relationship between depolarization, the activation of the membrane tryptophan carrier and the significance of this to the in vivo situation is discussed. PMID- 2965951 TI - Parastomal hernia in relation to site of the abdominal stoma. AB - Parastomal hernia is a common late complication of enterostomy, especially colostomy, and sometimes requires surgical treatment. A possible contributory factor, location of the stoma in relation to the rectus abdominis muscle, was studied by examination of 130 patients with permanent intestinal stoma. The bowel had been brought out through the rectus abdominis muscle in 107 patients and lateral to it in 23 patients. The respective prevalence of parastomal hernia in these groups was 2.8 per cent and 21.6 per cent. The highly significant difference indicates that enterostomy should be constructed through the rectus abdominis muscle, not lateral to it. PMID- 2965952 TI - Rash mediated by immune complexes associated with ranitidine treatment. PMID- 2965953 TI - Toilet aids. PMID- 2965954 TI - Neurotransmitter receptors in the avian brain. I. Dopamine receptors. AB - The distribution of dopamine D1 and D2 receptors was investigated in the pigeon brain using in vitro receptor autoradiography. D1 receptors were labeled in slide mounted tissue sections with [3H]SCH 23390. The agonist [3H]CV 205-502 and the antagonist [3H]spiroperidol were used to label D2 receptors. The highest densities of both D1 and D2 receptors were concentrated in the paleostriatum augmentatum, lobus paraolfactorius and olfactory tubercle. D1 receptors were in addition enriched in the entire pigeon telencephalon with exception of the ectostriatum. In contrast, no significant densities of D2 receptors were observed in the pallium. D2 but not D1 receptors were present in the tectum and nucleus pretectalis. The cerebellum presented D1 but not D2 receptors. These results demonstrate that in the avian brain, like in the mammalian one, both dopamine D1 and D2 receptors are concentrated in 'striatal' areas, while the telencephalon is rich in D1 but not D2 receptors. PMID- 2965955 TI - In vivo labeling of brain dopamine D2 receptors using the high-affinity specific D2 agonist [3H]CV 205-502. AB - The high-affinity selective dopamine D2 agonist [3H]CV 205-502 was utilized to in vivo label brain dopamine D2 receptors in the rat. Intravenous administration of [3H]CV 205-502 resulted in a selective accumulation of radioactivity in the striatum and in the pituitary. Smaller amounts of binding were found in the hypothalamus and cortex and non-significant binding was seen in the cerebellum. The binding of [3H]CV 205-502 was stereospecifically blocked by (+)-butaclamol but not by (-)-butaclamol. In vivo binding of [3H]CV 205-502 was also dose dependently blocked by other neuroleptics including sulpiride, haloperidol and spiroperidol, dopamine agonists such as bromocriptin, apomorphine and C1 201-678 but not by dopamine D1 antagonists or serotonin-2 antagonists. The regional distribution of the sites labeled in vivo by [3H]CV 205-502 was investigated by autoradiography and compared with the distribution seen after in vitro labeling in the consecutive sections, following washing of the label. The autoradiograms reveal the labeling of the same areas seen when in vitro autoradiography was used. High densities of binding were localized in the nucleus accumbens, striatum and olfactory tubercle as well as in the olfactory bulb. Lower densities were seen in the substantia nigra pars compacta and in the ventral tegmental area as well as in the stratum lacunosum molecular of the hippocampus and in the superior colliculus. The intermediate lobe of the pituitary also presented high densities while in the anterior pituitary only intermediate densities of receptor binding were observed. These results demonstrate that [3H]CV 205-502 is the first high affinity agonist useful for the autoradiographic visualization of dopamine D2 receptors after in vivo labeling. This compound could be modified for utilization in positron emission tomography imaging of dopamine D2 receptors in the living animal. PMID- 2965957 TI - Movement initiation characteristics in young adult rats in relation to the high- and low-affinity agonist states of the striatal D2 dopamine receptor. AB - Changes in the speed of movement initiation as a function of age, brain damage, or rat strain are associated with altered characteristics of nigrostriatal dopamine (DA) neurons and of striatal D2 DA receptors. In the present study we investigated the relationship between movement initiation (response parameters: percent of successful responses and response latency) and the agonist binding states of the D2 DA receptor in corpus striatum in 3-month-old Sprague-Dawley rats (n = 51). In contrast to the typical experimental procedure, the variances of the behavioral and receptor binding data were intentionally made as small as possible to provide the most stringent test of putative relationships among variables. Rats were trained to release a lever as rapidly as possible in response to a light/buzzer (CS) combination in order to avoid a mild footshock (UCS). Percent avoidance scores, latencies of the fastest successful trials (successful latencies) and mean latencies for all responses (mean latencies) were collected for 1000-, 500-, 300- and 200-ms CS-UCS intervals. Twenty-four hours following the last behavioral test, animals were euthanized for measurements of the high- and low-affinity binding of DA to D2 receptors in corpus striatum. The standard errors of the mean for both behavioral and receptor binding parameters were, generally, less than 10%. The tightness of the receptor binding data appeared to be related to a lack of biological variance in the animals rather than to an artifact associated with the behavioral testing procedure, since a parallel experiment indicated that different numbers of behavioral shaping sessions had no effects on striatal D2 binding characteristics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2965956 TI - 5-HT depletion with 5,7-DHT, PCA and PCPA in mice: differential effects on the sensitivity to 5-MeODMT, 8-OH-DPAT and 5-HTP as measured by two nociceptive tests. AB - Depletion of 5-hydroxytryptamine (5-HT) in mice was produced by intracerebroventricular injection of 5,7-dihydroxytryptamine (5,7-DHT, 80 micrograms) or by systemic injections of p-chloroamphetamine (PCA, 3 X 40 or 4 X 40 mg/kg), p-chlorophenylalanine (PCPA, 5 X 400 or 14 X 400 mg/kg) or combined PCA (3 X 40 mg/kg) + PCPA (11 X 400 mg/kg). Neither of the pretreatments altered nociception in the increasing temperature hot-plate test, whereas hyperalgesia was demonstrated in 5,7-DHT lesioned animals in the tail-flick test. 5,7-DHT pretreatment enhanced the antinociceptive effect of the 5-HT agonists 5-methoxy N,N-dimethyltryptamine (5-MeODMT), 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH DPAT) and 5-hydroxytryptophan (5-HTP). This effect was observed after 2, 5 and 8 days in the tail-flick test and after 5 and 8 days in the hot-plate test. However, pretreatment with PCPA or PCA failed to alter the antinociception elicited by the 5-HT agonists, although a tendency towards enhancement of antinociception was found after combined treatment with PCA and PCPA. It is suggested that the injection of 5,7-DHT induces denervation supersensitivity of post-synaptic 5-HT receptors. The lack of such supersensitivity after PCPA pretreatment which induces similar 5-HT depletion to 5,7-DHT, may suggest that other factors than the absence of 5-HT may contribute to the development of denervation supersensitivity. Alternatively, the three 5-HT depleting agents may produce a qualitatively different reduction of 5-HT. PMID- 2965958 TI - Striatal D1 dopamine receptor distribution following chemical lesion of the nigrostriatal pathway. AB - The morphochemical and biochemical distribution of the adenylate cyclase-linked dopamine receptor, or D1 subpopulation, has been examined in the rat striatum following a chemical lesion of the dopaminergic nigrostriatal pathway. The cellular pattern of D1 dopaminergic binding was assessed using in vitro autoradiographic localization of [3H]SCH 23390 or [125I]SCH 23982, selective D1 receptor antagonists, 1 week following unilateral infusion of the neurotoxin 6 hydroxydopamine (6-OHDA) into the substantia nigra. The specific association of the D1 binding sites with cyclic AMP-immunoreactive striatal neurons was abolished after lesion of the dopaminergic nigral afferents. The morphochemical disruption of the caudate D1 dopamine binding sites in relation to cyclic AMP positive elements, a large proportion of which are striatonigral neurons, probably contributes to the dysfunctions in this subpopulation of dopamine receptor after depletion of the catecholamine neurotransmitter. PMID- 2965960 TI - Muscle spindle number is normal in guinea pigs with reduced dorsal root ganglia following in utero deprivation of nerve growth factor. AB - Muscle spindles are induced during embryonic development by contact between sensory axons and undifferentiated myotubes. Neonatal guinea pigs deprived of nerve growth factor in utero can have only 15-20% of the normal number of dorsal root ganglion cells. This study demonstrates that such neonates have normal numbers of muscle spindles. The remaining 15-20% of dorsal root ganglion cells may therefore largely be muscle afferents. PMID- 2965959 TI - Atrial natriuretic polypeptide depresses angiotensin II induced excitation of neurons in the rat subfornical organ in vitro. AB - The effects of atrial natriuretic polypeptide (ANP) on the extracellularly recorded activity of neurons in the subfornical organ (SFO) were investigated in rat brain slice preparations by adding the peptide to the perfusion medium. Eight (14%) of 56 SFO neurons were inhibited and none of the cells were excited by ANP at 10(-7) M. Of 42 SFO neurons tested with both ANP and angiotensin II (AII) at 10(-7) M, 4 (10%) cells were inhibited by ANP and excited by AII, the remainder responded to either one or other of the peptides but not both or were unresponsive. In 13 (87%) of 15 SFO neurons, ANP at 10(-7) M depressed by more than 40% the excitation induced by AII at 10(-7) M, while ANP did not always depress the excitation induced by raising the extracellular potassium concentrations in 6 SFO cells tested. We conclude that ANP strongly depresses AII induced excitation in all SFO neurons, although it has very weak inhibitory effects on spontaneous activity, thus ANP may act as a neuromodulator in the SFO. PMID- 2965962 TI - [Clinical significance of atrial natriuretic factor in primary hyperaldosteronism, Cushing syndrome and pheochromocytoma]. PMID- 2965961 TI - Disinhibition of nucleus accumbens neurons by the dopamine D2 receptor agonist LY 141865: prevented by 6-OHDA pretreatment. AB - Intravenous administration of LY-141865, a specific dopamine (DA) D2 receptor agonist, caused a biphasic increase/decrease in the firing rate of cells in the rat nucleus accumbens (NAc). 6-Hydroxydopamine injected directly into the medial forebrain bundle, through which the ascending DA fibers pass, prevented the initial rate increase of NAc neurons produced by low doses of LY-141865. In contrast, pretreatment of rats with 5,7-dihydroxytryptamine did not alter the biphasic response of NAc neurons to LY-141865. These results, together with previous findings that D2 autoreceptors are much sensitive to DA and DA agonists, strongly support the hypothesis that the initial rate-increase caused by low doses of LY-141865 is the result of disinhibition. PMID- 2965963 TI - [125I-BH-8-MeO-N-PAT, a new ligand for the study of 5-HT1A receptors in the central nervous system]. AB - Specific radioactive ligands are needed for studying the pharmacological properties and the regional distribution of the different classes of 5-HT1 receptors within the central nervous system. We describe here the synthesis and some characteristics of the first iodinated specific ligand of 5-HT1A receptors. Like its parent compound, the agonist 8-hydroxy-2-(di-n-propylamino)tetralin or 8 OH-DPAT, [125I]-BH-8-MeO-N-PAT, exhibits a high affinity and excellent selectivity for 5-HT1A sites. Its high specific radioactivity makes this ligand a useful tool for studying 5-HT1A receptors in membranes and sections of the rat brain. PMID- 2965965 TI - Reperfusion hemorrhage following PTCA and thrombolysis for left main coronary artery occlusion. AB - Myocardial reperfusion hemorrhage has been reported in humans but rarely after percutaneous transluminal coronary artery angioplasty (PTCA) of occluded vessels and not after the reopening of left main occlusions. Three patients with infarct and cardiogenic shock and reperfusion hemorrhages after PTCA, with or without streptokinase infusion and reopening of acutely thrombosed left main coronary arteries are reported here. The effect of the reperfusion 24,9 and 7 h after symptom onset and 24,25 and 3 h before death is examined. PMID- 2965964 TI - Local morphologic effects of coronary artery balloon angioplasty. AB - The morphologic effects of percutaneous transluminal coronary artery balloon angioplasty (PTCA) on atherosclerotic vessels is described in six patients who died at varying intervals after the procedure (four early and two late). In the early group (less than one week post PTCA) one patient died because of electromechanical dissociation during emergency PTCA for evolving infarct; in the three other patients PTCA was performed for left main occlusion and cardiogenic shock with deaths 3, 24 and 25 h after PTCA. The two late deaths were patients who died one and nine months after PTCA from unrelated causes. There were 12 sites of balloon inflation in the six patients, all in left main, isolated marginal or left anterior descending arteries. Post mortem examinations, with in toto serial sectioning of the ballooned coronary arteries, revealed a number of local morphologic changes. Plaque fractures and disruptions of the arterial wall to variable depths were observed. At four sites these fractures were through media, and at one site was associated with a large dissection. These cases had only small epicardial hemorrhages or reactive adventitial changes associated with these deep fractures. Four of the six patients had intramural arterial emboli (athero/thrombo/calcium/foreign body). These findings confirm that a large part of the effect of PTCA is due to physical disruption of plaque and underlying native vessel. PMID- 2965966 TI - CT scanography for limb length determination. AB - The authors present an alternative to classic techniques used to measure limb length discrepancy radiographically. CT scanography seems to have advantages over currently-used Bell-Thompson roentgenography in that it uses less radiation and is of no increase in cost. PMID- 2965967 TI - Treatment of tardive dyskinesia: preliminary report on use of tetrabenazine. AB - Tetrabenazine has been used for treatment of tardive dyskinesia sporadically over the past twenty years. Dose has usually been decided empirically without assaying blood levels. This report describes 23 cases treated successfully with tetrabenazine. Our method of measuring levels of tetrabenazine and its metabolites in biological samples is described briefly. PMID- 2965968 TI - Effects of diphosphonate and x-rays on bone lesions induced in rats by prostate cancer cells. AB - Prostate adenocarcinoma-III (PA-III) cells deposited over the scapula of Lobund Wistar (L-W) rats induced osteolytic and osteoplastic changes in that bone. The osteolytic process was prevented and interrupted by administrations of dichloromethylene diphosphonate (Cl2MDP); but the osteoplastic process, once initiated, continued. The latter process was interrupted by ionizing radiation. Treatments of PA-III-affected bones with Cl2MDP and x-rays immobilized both osteolytic and osteoplastic processes. PMID- 2965969 TI - Induction of ovarian granulosa cell tumors in SWXJ-9 mice with dehydroepiandrosterone. AB - Spontaneous ovarian granulosa cell (GC) tumors develop in SWXJ-9 inbred mice at approximately the time of puberty. The effect of dehydroepiandrosterone (DHEA), a steroid secreted by the adrenals and reported to have antitumor actions, was examined in this ovarian tumor model. In contrast with expectations, administration of diet supplemented with 0.4% DHEA or Silastic capsules containing 10 mg DHEA resulted in a significant multifold increase in GC tumor incidence. Similar studies with metabolites of DHEA, i.e., testosterone (TESTO), dihydrotestosterone (DHT), and 17 beta-estradiol (E2), revealed that TESTO was as effective as DHEA in increasing GC tumor incidence. DHT was without effect, and E2 suppressed GC tumor incidence. Serum steroid levels and steroid target tissue responses were assessed to determine if a correlation between a change in level or response to specific steroids and GC tumorigenesis existed. In both tumor-free and GC tumor host mice, dietary or capsular treatment with DHEA, TESTO, or DHT resulted in substantial alteration in one or more of serum steroids, DHEA, androstenedione, TESTO, and DHT, in addition to the administered steroid. No consistent correlation was observed between changes in a single steroid or pattern of steroids and GC tumorigenesis. Although significant increases in serum estrogens could be detected in GC tumor hosts treated with DHEA but not TESTO, estrogens did not induce these tumors. Treatment with E2 increased only serum E2 levels. In tumor-free mice, DHEA and E2 treatments were associated with vaginal cytological evidence of estrogen action, whereas the androgens induced a leukocytic pattern. Eighty-eight % of GC tumor host mice, regardless of steroid treatment, showed a vaginal cytology pattern that included cornified cells. The evidence presented in this report leads us to hypothesize that (a) spontaneous and steroid-induced GC tumorigenesis in these mice have the same mechanism, and (b) subtle increases in DHEA or a closely related metabolite during the peripubertal period may initiate GC tumors in these genetically susceptible mice. The mechanism whereby these steroids initiate GC tumorigenesis remains to be determined. PMID- 2965970 TI - Fluorodeoxyuridine modulation of the incorporation of iododeoxyuridine into DNA of granulocytes: a phase I and clinical pharmacological study. AB - The amount of iododeoxyuridine (IdUrd) incorporated into DNA determines the degree of radiosensitization. Fluorodeoxyuridine (FdUrd) has been shown to biochemically modulate IdUrd incorporation into DNA in vitro and in vivo. In this Phase I study, these drugs were coadministered to patients during 14-day continuous i.v. infusion periods in order to investigate whether the incorporation of IdUrd into DNA in vivo could be increased without increasing the dose of IdUrd. IdUrd plasma concentrations and incorporation of IdUrd into DNA of granulocytes were measured by high-performance liquid chromatography. Up to 8.8% substitution of thymidine by IdUrd was observed. Even at 3.5 mg/m2/day FdUrd for 14 days (78% of the maximum-tolerated dose as a single agent), no clinically relevant enhancement of incorporation of IdUrd into DNA of granulocytes was observed. Also, no changes in plasma levels of IdUrd were observed with escalating doses of FdUrd. Toxicity patterns (stomatitis, diarrhea, and bone marrow depression) and isobologram analysis suggested that IdUrd and FdUrd had additive, rather than synergistic, effects. PMID- 2965971 TI - The branched malto-oligosaccharides resulting from the action of Bacillus macerans cycloamylose glucanotransferase on 6-O-alpha-D-glucopyranosyl cyclomaltohexaose plus D-glucose. PMID- 2965972 TI - Suppressor deletion therapy: selective elimination of T suppressor cells in vivo using a hematoporphyrin conjugated monoclonal antibody permits animals to reject syngeneic tumor cells. AB - A MAb (B16G) which recognizes a constant epitope on TsC and their soluble factors in DBA/2 mice has been described previously. In this study, we show that when this MAb is covalently linked to the photoactivable molecule Hp, and injected i.v. into P815 tumor-bearing mice which were subsequently exposed to light, tumors undergo permanent regression in 10%-40% of these mice (depending on the individual experiment). All control animals died within an average of 22-24 days after tumor cell injection. It is suggested that tumor regression is attributable to immune mechanisms facilitated by the elimination of a population of TsC. When splenocytes of B16G-Hp-treated mice were assayed in vitro for the generation of CTL active against P815 tumor cells, it was found that 24 h after treatment, a significant increase in killer cell activity was noted but that this effect was gone by 48h. We also show that B16G-Hp conjugates are capable in vitro of specifically killing cells of a TsC hybridoma, A10 (which has been shown previously to secrete a T suppressor factor reactive with P815 cell surface antigens). This conjugate had no cytotoxic effect on P815 cells under conditions in which A10 cells were killed. PMID- 2965973 TI - Induction by an immunogenic immunomodulating agent of nonspecific T cell suppression of lymphocyte responsiveness in MLR but not of antibody production. AB - Spleen cells derived from BALB/c mice that had been repeatedly immunized with the methanol extraction residue (MER) fraction of tubercle bacilli exhibited a depressed capacity to act as responder cells in allogeneic and syngeneic mixed lymphocyte reactions (MLR). Previously reported studies revealed that such spleen cells are also defective in the in vitro generation of antibodies. In order to determine the nature of the cells responsible for the depressed MLR reactivity, purified populations of splenic macrophages, B lymphocytes, T lymphocytes originating from normal and from MER-immunized mice, and cell culture supernatants were added to MLR mixtures consisting of normal mouse splenocytes. Macrophages originating from MER-immunized mice and their culture supernatants exerted a significantly higher suppressive effect on MLR than that of corresponding preparations from normal mice. Splenic T cells originating from MER immunized mice and their supernatants also significantly suppressed the MLR response. However, the same T cell populations that were inhibitory in MLR failed to suppress the in vitro generation of antibodies against sheep red blood cells in the presence of either MER or 2-mercaptoethanol. These and previously reported findings indicate that a nonspecific immunomodulating agent, MER, can, under certain conditions of treatment, elicit the induction of nonspecific suppressor T cells for MLR but not for antibody production, and, accordingly, can inhibit cellular and humoral immunological responsiveness by different mechanisms. PMID- 2965974 TI - [The duplex scanner]. PMID- 2965975 TI - [Thallium-201 scintigraphy in the early and non-invasive diagnosis of obliterative arteriopathies of the lower limbs]. PMID- 2965976 TI - [Scintigraphy with thallium-201 in the evaluation of vasoactive treatment in subjects with obliterative arteriopathies of the lower limbs]. PMID- 2965977 TI - [Coronary angioplasty: an alternative to heart surgery?]. PMID- 2965978 TI - [Atrioseptostomy, angioplasty, valvuloplasty]. PMID- 2965979 TI - [Laboratory of hemodynamics today: current status and perspectives in Italy]. PMID- 2965980 TI - [Peripheral dopamine DA1-type receptors: their localization, functional role and possible clinical implications]. PMID- 2965981 TI - [DA2-type dopamine receptors: the physiological and clinical implications in cardiology]. PMID- 2965982 TI - [Atrial natriuretic peptides and essential arterial hypertension: considerations on present knowledge and possible trends in research]. PMID- 2965983 TI - [Characteristics of coronary circulation in left ventricular hypertrophy secondary to aortic valve disease and dilated cardiomyopathy]. PMID- 2965984 TI - [Protective effect of verapamil in myocardial hypertrophy due to aortic stenosis experimentally induced in the rat]. PMID- 2965986 TI - AIDS and the Chicago dentist shine national spotlight on CDS. PMID- 2965985 TI - Target immunity of Mu transposition reflects a differential distribution of Mu B protein. AB - A DNA molecule carrying Mu end DNA sequence(s) is a poor target in the Mu DNA strand-transfer reaction, a phenomenon which is referred to as "target immunity." We find that Mu B protein stimulates intermolecular strand-transfer by binding to the target DNA. Our results show that a differential distribution of Mu B protein between "immune" and "non-immune" DNA molecules is responsible for target immunity; in the presence of Mu A protein and ATP, Mu B protein dissociates preferentially from immune DNA molecules. Hydrolysis of ATP is implicated in establishing the differential distribution of Mu B protein between immune and non immune DNA molecules in the presence of Mu A protein; nonhydrolyzable ATP gamma S can support an efficient strand-transfer reaction even with a target DNA that is immune in a reaction with ATP. PMID- 2965988 TI - Differential effects of cyclosporins A and G on functional activation of a T helper-lymphocyte line mediating experimental autoimmune uveoretinitis. AB - The effect and relative efficiency of cyclosporin A (CsA) and cyclosporin G (CsG) on suppressing the activation of primed autoimmune rat T-helper lymphocytes were assayed. The autoimmune T-helper cells (ThS) are a long-term line specific to the retinal soluble antigen (SAg) and can adoptively transfer experimental autoimmune uveoretinitis (EAU), after in vitro reactivation with antigen or mitogen, to naive syngeneic hosts. Antigen-driven production of interleukin-2 (IL-2) and antigen-driven proliferation were inhibited in a dose-dependent manner and to a similar extent at each of the respective cyclosporin concentrations. CsA was 8-10 times more potent than CsG, with ID50-CsA occurring at 0.5 to 2 ng/ml, and ID50 CsG at 5 to 20 ng/ml, depending on the experiment and the cyclosporin batch. Addition of exogenous lymphokines in the form of rat spleen concanavalin A (Con A)-conditioned medium (SCM) or recombinant IL-2 (but not recombinant IL-1) was able to reverse only about half of the inhibition, as measured along the linear part of the dose-response curve. Inhibition of IL-2 production was lost if a maximally inhibitory dose of cyclosporin was added to the cultures later than 8 hr after antigen stimulation, while proliferation was still suppressed to 50% by cyclosporin added as late as 12 hr and could not be restored by addition of SCM. Both cyclosporins at concentrations that blocked proliferation and IL-2 production significantly suppressed the generation of high-affinity and low affinity IL-2 receptors by ThS in response to antigen (as assayed by direct binding of 125I-IL-2). These results suggest that CsA and CsG inhibit antigen induced expansion of ThS by interfering with more than one activation step. In contrast, the in vitro activation of the uveitogenic potential of ThS cells, incubated with antigen in the presence of CsA or CsG and adoptively transferred into untreated recipients, was not affected by the cyclosporins. Thus, triggering of the pathogenic potential of primed autoimmune T-helper lymphocytes can take place in the presence of cyclosporin and in the absence of cellular proliferation. PMID- 2965987 TI - Different signals for stimulation of proliferation and lymphokine secretion by a CD3+ WT31- cloned cytotoxic lymphocyte. AB - CD3+ WT31- T cells were sorted from peripheral blood of a normal healthy donor by a FACS IV and cloned by limiting dilution in the presence of a phorbol ester (tetradecanoyl phorbol acetate, TPA), calcium ionophore (ionomycin, Io), interleukin-2 (IL-2), allogeneic cells, and phytohemagglutinin (PHA). One of the derived clones, 290-2, was investigated in detail. 290-2 mediated strong natural killer (NK) but not lymphokine-activated killer (LAK) activity. It proliferated in the presence of IL-2 but not IL-4. It carried the surface phenotype CD3+ WT31- CD4weak+ CD8-, CD16-, and Leu 19+. Expression of CD4 was heterogeneous within the clone, since two of three subclones were also CD4weak+ but one was CD4-. NK activity was blocked by monoclonal antibody (moAb) to CD1 1a (LFA1), but not by monoclonal antibody (moAb) to either CD3 or CD4. Northern blotting revealed T cell receptor (TCR-gamma) but not alpha- or full-length beta-chain mRNA. 290-2 proliferated autonomously when stimulated with a combination of TPA +Io, with PHA or CD3 moAb and autologous B-cell lines (B-LCL) (and this was inhibited by an anti-IL-2 receptor moAb), but not to allogeneic B-LCL or any of the other stimulating agents alone. Unexpectedly, the TPA + Io stimulus which resulted in maximal proliferative responses did not trigger interferon-gamma or granulocyte/macrophage colony-stimulating factor production, although both lymphokines were secreted in the presence of B-LCL + TPA + Io. Proliferative responses were not enhanced by the presence of B-LCL. Thus, activation signals sufficient for autocrine proliferative responses were insufficient for secretion of other lymphokines. Such clones will provide valuable reagents for investigating the biology of the TCR-gamma+ T cell. PMID- 2965989 TI - Nicotinamide protects target cells from cell-mediated cytolysis. AB - Nicotinamide in concentrations of 5 mM and greater protected fibroblast target cells from lysis by lymphokine-activated killer cells (LAK cells). Protection was concentration dependent and was exerted at the level of the target cell. Nicotinamide did not interfere with effector-target cell conjugate formation or with the calcium dependent triggering step of the lytic process. Target cell lysis in cultures without nicotinamide was accompanied by fragmentation of target cell DNA. The DNA of target cells cultured with LAK cells in the presence of nicotinamide remained intact. 3-Aminobenzamide which, like nicotinamide, inhibits poly(ADP-ribose) synthetase but is not a precursor of NAD, was an effective inhibitor of target cell lysis while nicotinic acid, an alternative precursor of NAD in cells, was not. The data point to a central role for poly(ADP-ribose) synthetase in the events leading up to DNA fragmentation and the release of 51Cr from target cells damaged by lymphokine-activated killer cells. PMID- 2965990 TI - Immature CD4- CD8+ murine thymocytes. AB - Mature thymocytes are usually defined and separated from other less mature thymocytes on the basis of their mutually exclusive expression of either CD4 or CD8. However, such murine "single positives" include a subpopulation of immature cells with properties resembling CD4- CD8- thymocytes or CD4+ CD8+ cortical blasts. Most of these immature single positives are CD4- CD8+, some expressing relatively low levels of CD8. They are large, dividing cortisone-sensitive cells found in the outer cortex. They express high levels of the heat-stable antigen (recognized by the monoclonals M1/69, B2A2, and J11d) but they are MEL-14-. The absence of detectable surface CD3, the absence of alpha-chain messenger RNA, and the predominance of the truncated form of the beta-chain messenger RNA all indicate that they do not express the T-cell antigen-receptor complex. Strategies for eliminating such immature cells from preparations of mature thymocytes are given, and their developmental significance is discussed. PMID- 2965992 TI - Light cured composites. PMID- 2965991 TI - Generation of suppressor T cells after local immunization with histocompatibility antigens. AB - Subcutaneous (sc) hind-foot immunization (HFI) of mice with allogeneic spleen cells can induce a state of delayed-type hypersensitivity (DTH) as well as a state of suppression of DTH. This paper deals with the suppression induced by HFI. The state of suppression could be adoptively transferred by spleen cells and lymph node cells between Days 3 and 7 after HFI only. However, in the hind-foot immunized mice the state of suppression lasted at least 25 days. The suppressor cells expressed the Thy-1+, Lyt-1-2+ phenotype and suppressed DTH antigen specifically. The suppressor cells, however, also suppressed DTH responses to unrelated third-party alloantigens, provided the latter were administered during the induction of DTH together with the same alloantigens that were used for HFI. The HFI-induced T-suppressor cells suppressed the induction phase of DTH (i.e., the proliferative activity of the draining lymph node cells after secondary sc immunization), but not the expression phase of DTH (i.e., the activity of previously activated DTH effector T cells). H-2D compatibility between the donors of the HFI-induced T-suppressor cells and the recipients was required for the adoptive transfer of suppression. The differences in effect of local immunization versus systemic immunization on the induction and functional activity of T suppressor cells are discussed. PMID- 2965993 TI - Interaction of Chlamydomonas dynein with tubulin. AB - Studies were conducted to determine if dynein could bind to unpolymerized tubulin. Tubulin alone normally fractionated in the included volume of a molecular sieve Bio-Gel A-1.5m column. Incubated together, tubulin and dynein coeluted in the void volumn, suggesting that a complex had formed between the two. In addition, immunoelectron microscopy revealed preassembled microtubules were labeled with biotin antibody only when incubated in both dynein and biotinylated tubulin, evidence that dynein with bound biotinylated tubulin had decorated the microtubules. A fraction of the tubulin could be dissociated from dynein by addition of ATP and vanadate, as assayed by molecular sieve chromatography followed by densitometry of gels, suggesting that some tubulin bound to the B end of the dynein arm. Additional tubulin dissociated from the dynein under conditions of high salt. These studies, together with those indicating that tubulin blocked the A end of the dynein arm from binding to microtubules and promoted the interaction of two arms at their A ends, provide evidence that the A end of the arm also can bind tubulin. Thus, the tubulin subunits, themselves, on a microtubule rather than a particular surface lattice structure formed by adjacent protofilaments may provide the binding sites for both ends of the dynein arm. PMID- 2965994 TI - A monoclonal antibody to the Ca2+-ATPase of cardiac sarcoplasmic reticulum cross reacts with slow type I but not with fast type II canine skeletal muscle fibers: an immunocytochemical and immunochemical study. AB - Ca2+-ATPase of the sarcoplasmic reticulum was localized in cryostat sections from three different adult canine skeletal muscles (gracilis, extensor carpi radialis, and superficial digitalis flexor) by immunofluorescence labeling with monoclonal antibodies to the Ca2+-ATPase. Type I (slow) myofibers were strongly labeled for the Ca2+-ATPase with a monoclonal antibody (II D8) to the Ca2+-ATPase of canine cardiac sarcoplasmic reticulum; the type II (fast) myofibers were labeled at the level of the background with monoclonal antibody II D8. By contrast, type II (fast) myofibers were strongly labeled for Ca2+-ATPase of rabbit skeletal sarcoplasmic reticulum. The subcellular distribution of the immunolabeling in type I (slow) myofibers with monoclonal antibody II D8 corresponded to that of the sarcoplasmic reticulum as previously determined by electron microscopy. The structural similarity between the canine cardiac Ca2+-ATPase present in the sarcoplasmic reticulum of the canine slow skeletal muscle fibers was demonstrated by immunoblotting. Monoclonal antibody (II D8) to the cardiac Ca2+-ATPase binds to only one protein band present in the extract from either cardiac or type I (slow) skeletal muscle tissue. By contrast, monoclonal antibody (II H11) to the skeletal type II (fast) Ca2+-ATPase binds only one protein band in the extract from type II (fast) skeletal muscle tissue. These immunopositive proteins coelectrophoresed with the Ca2+-ATPase of the canine cardiac sarcoplasmic reticulum and showed an apparent Mr of 115,000. It is concluded that the Ca2+ ATPase of cardiac and type I (slow) skeletal sarcoplasmic reticulum have at least one epitope in common, which is not present on the Ca2+-ATPase of sarcoplasmic reticulum in type II (fast) skeletal myofibers. It is possible that this site is related to the assumed necessity of the Ca2+-ATPase of the sarcoplasmic reticulum in cardiac and type I (slow) skeletal myofibers to interact with phosphorylated phospholamban and thereby enhance the accumulation of Ca2+ in the lumen of the sarcoplasmic reticulum following beta-adrenergic stimulation. PMID- 2965995 TI - Amber nonsense mutations in regulatory and structural genes of the nitrogen control circuit of Neurospora crassa. AB - Neurospora crassa possesses a set of nitrogen-regulated enzymes whose expression requires a lifting of nitrogen catabolite repression and specific induction. The nit-2 gene is a major regulatory locus which appears to act in a positive way to turn on the expression of these nitrogen-related enzymes whereas the nit-4 gene appears to mediate nitrate induction of nitrate and nitrite reductase. The nit-3 gene specifies nitrate reductase and is subject to control by both nit-2 and nit 4. Many new nit-2, nit-3, and nit-4 mutants were isolated in order to obtain amber nonsense mutations in these loci which were suppressible by the suppressor gene, Ssu-1. A nit-2 nonsense mutant was isolated which has altered regulatory properties for control of nitrate reductase. L-amino acid oxidase, and uricase, and which may encode a truncated regulatory protein. Four nit-3 nonsense mutations were isolated, each of which completely lacks nitrate reductase activity, which is restored to markedly different levels by suppression with Ssu 1. Studies of heat activation and thermal lability of nitrate reductase suggest a qualitative alteration of the enzyme occurs in two of the Ssu-1 nit-3 strains. PMID- 2965996 TI - Construction and physiological characterization of mutants disrupted in the phosphofructokinase genes of Saccharomyces cerevisiae. AB - The structural genes coding for the two kinds of subunits of phosphofructokinase in yeast have been cloned previously (Heinisch 1986). The coding regions were defined by S1-mapping. They were disrupted in vitro by insertion of a LEU2 marker. These constructions were then used for substitution of the respective chromosomal copies. That the disruption of the PFK-genes had in fact occurred was confirmed by Southern blot analysis. Furthermore, in Northern blots shorter transcripts were detected in the respective disruption mutants. Using polyclonal antibodies the alpha-subunits were not detectable in pfk1-disruptions whereas the beta-subunits were undetectable in pfk2-disruptions. Physiological characterization showed that the single disruption mutants still fermented glucose to ethanol and CO2. They accumulated fructose-6-phosphate and glucose-6 phosphate over wild type levels and showed decreased levels of fructose-1,6 bisphosphate. In addition an accumulation of sedoheptulose-7-phosphate was observed, a metabolite not detectable in wild type cells. A haploid yeast strain containing both disrupted copies of the PFK-genes is not capable of growing on rich medium containing 2% glucose. The accumulation of glucose-6-phosphate, fructose-6-phosphate and sedoheptulose-7-phosphate is much more pronounced in such mutants, whereas the fructose-1,6-bisphosphate concentration decreases below the level of detection. PMID- 2965998 TI - [Evaluation of post-traumatic spinal conditions]. PMID- 2965997 TI - [Comparison of findings in hypertrophy of the ventricular myocardium in rats after administration of thyroxine and ligation of the aorta]. PMID- 2965999 TI - [What should the pediatrician know about social service benefits for handicapped children and their families? I]. PMID- 2966000 TI - [Radionuclide uroflowmetry and ureteral measurement in girls with diseases of the urinary tract]. PMID- 2966001 TI - [Studies on racial difference in distribution of apoE phenotypes and frequencies of apoE alleles--comparison of Chinese and Western populations]. PMID- 2966002 TI - [Expression of seven oncogenes in HL-60-AR cells and their changes after chemically induced differentiation]. PMID- 2966003 TI - [Studies on the effect of the combined use of dl-15methyl-prostaglandin F2 alpha methyl ester with diphenoxylate hydrochloride in early pregnancy in rats]. PMID- 2966004 TI - [Nucleic acid hybridization analysis of the regulatory effect of the cytoplasmic factor on the malignancy of myeloma cells. II. Expression of the myc oncogene in homo- and hetero-cellular hybridization]. PMID- 2966005 TI - [Effects of sodium molybdate on sister chromatid exchange in V79 cells induced by MNU and MNNG]. PMID- 2966006 TI - [Effects of poriatin on mouse peritoneal macrophages]. PMID- 2966007 TI - [Effects of lung volume and alveolar surface tension on pulmonary vascular resistance]. PMID- 2966008 TI - [Experimental study on the quantification of the invasive process of tumor cells in vivo using a computer-image processing system]. PMID- 2966009 TI - [Computer application in DNA sequence analysis of hepatitis B virus]. PMID- 2966010 TI - [Serological and histopathological studies on viral hepatitis of the Citellus dauricus]. PMID- 2966012 TI - [Rapid microbiological identification system for Enterobacteriaceae]. PMID- 2966011 TI - [Tripterygium wilfordii in the treatment of the nephritis of anaphylactoid purpura]. PMID- 2966013 TI - [Experimental studies on transvenous catheter endomyocardial biopsy in dogs]. PMID- 2966014 TI - Sexual maturation in girls with severe mental handicap. AB - It is becoming accepted that sex education of handicapped adolescents should not be neglected. For such education to be effective, it should begin before sexual maturity. In an attempt to provide a starting guideline for girls attending ESN(S) schools in the Manchester area, a cross-sectional survey was performed to determine mean age of menarche. The results showed the group as a whole to be on average 13 months later than normal. However, Down's syndrome formed a large identifiable subgroup with a mean menarcheal age 11 months earlier than normal, leaving the non-Down's group 21 months later than normal, and 2 years 8 months behind the Down's group. The implications for education are discussed. PMID- 2966015 TI - Augmented expression of atrial natriuretic polypeptide gene in ventricles of spontaneously hypertensive rats (SHR) and SHR-stroke prone. AB - Tissue levels of atrial natriuretic polypeptide (ANP) and ANP messenger RNA (mRNA) in the atrium and ventricle were measured simultaneously in spontaneously hypertensive rats (SHR) and its substrain, SHR-stroke prone (SHRSP), and these levels were compared with those in control Wistar-Kyoto rats (WKY). At 27 weeks of age with established hypertension and ventricular hypertrophy, ANPmRNA levels in ventricles from SHR and SHRSP were markedly increased, and total contents of ventricular ANPmRNA in SHR and SHRSP were approximately 50% and 250%, respectively, of the corresponding atrial contents. However, levels and total contents of atrial ANPmRNA in SHR and SHRSP were similar to those of WKY, and the total content of ventricular ANPmRNA in WKY was only 6% of the content of atrial ANPmRNA. ANP concentrations in ventricles of SHR and SHRSP were increased in association with the augmentation of ANPmRNA levels. During the prehypertensive stage at 6 weeks of age, slight increases in levels and total contents of ANPmRNA and ANP in the ventricle were observed only in SHRSP. These results demonstrate that the expression of the ANP gene is markedly augmented in ventricles of SHR and SHRSP, especially of SHRSP, at the stage of established hypertension and ventricular hypertrophy, and they also suggest that these genetically hypertensive rats are one of the best animal models to investigate the biosynthesis, storage, and secretion of ventricular ANP. PMID- 2966016 TI - Characteristics and outcome of patients in whom reperfusion with intravenous tissue-type plasminogen activator fails: results of the Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) I trial. AB - To examine the outcome of patients with persistent coronary artery occlusion despite treatment with intravenous tissue-type plasminogen activator (t-PA), the clinical course of 96 patients with persistent occlusion after 90 min of therapy was evaluated. All patients underwent cardiac catheterization 90 min after initiation of intravenous t-PA. Immediate coronary angioplasty (PTCA) was attempted when the infarct-related artery failed to reperfuse unless the vessel was technically unsuitable or the infarct was thought to be small. No baseline differences could be found between these 96 patients and 288 patients who achieved perfusion with the same protocol. Although patients with and without successful perfusion after t-PA had similar clinical courses before cardiac catheterization, those without perfusion had more complications (ventricular fibrillation, severe bradycardia, hypotension) during catheterization. PTCA achieved reperfusion with less than 50% residual stenosis in 73% of the 86 patients in whom it was attempted, while 16% were left with a high-grade (greater than 50%) residual stenosis and PTCA failed in 11%. Mortality was highest in the nine patients with complete PTCA failure (44%), compared with a 6% mortality in the 63 patients with an insignificant residual stenosis after PTCA and a 14% mortality in the 14 patients with reperfusion, but a greater than 50% residual stenosis after PTCA. In 10 patients with small infarcts (six), unsuitable anatomy (two), or "spontaneous" drug induced (but later) opening before contemplated PTCA (two), PTCA was not attempted and no mortality occurred. The in-hospital reocclusion rate after successful PTCA was 29%, despite the use of heparin and antiplatelet agents.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966018 TI - The effect of hypertension and left ventricular hypertrophy on the lower range of coronary autoregulation. AB - These studies were performed to test the hypothesis that left ventricular hypertrophy arising as a complication of chronic hypertension is associated with impaired coronary autoregulation. Twelve dogs with hypertension and left ventricular hypertrophy (one-kidney, one-clip model) and 11 normal dogs were instrumented and subsequently studied while conscious. Circumflex pressure, measured with an intracoronary catheter, was adjusted to 100, 75, and 40 mm Hg with a hydraulic occluder that was placed proximally. At each circumflex pressure, myocardial perfusion was measured with radioactive microspheres. Reduction of circumflex pressure over this range did not significantly alter heart rate, left atrial pressure, or arterial pressure. In normal dogs, reduction of circumflex pressure did not alter total myocardial perfusion or the transmural distribution of perfusion. In contrast, in dogs with hypertension and left ventricular hypertrophy, circumflex subendocardial perfusion decreased 46% when pressure was decreased from 100 to 40 mm Hg (p less than .05 compared with normal). Autoregulation was quantified for each third of myocardium with the use of autoregulatory gain values (1 = perfect autoregulation; 0 = the absence of autoregulation). For pressure changes of 100 to 75 mmHg, values for autoregulatory gain were near unity for all layers of myocardium in both groups of animals. When pressure was decreased from 75 to 40 mm Hg, values for autoregulatory gain among the normal and hypertensive groups were, respectively: for subepicardium 1 +/- 0.2 (mean +/- SE) vs 0.9 +/- 0.2 (p = NS), for the midwall 0.8 +/- 0.2 vs 0.5 +/- 0.2 (p = NS), and for the subendocardium 0.8 +/- 0.1 vs 0.1 +/- 0.2 (p less than .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966017 TI - Coronary arterial thrombolysis with combined infusion of recombinant tissue-type plasminogen activator and urokinase in patients with acute myocardial infarction. AB - To determine whether tissue-type plasminogen activator (t-PA) and urokinase (UK) act synergistically to achieve coronary thrombolysis, incremental doses of both drugs were infused intravenously over 60 min. In 146 consecutive patients treated 3.0 +/- 1.0 hr from symptom onset, coronary angiography was performed 90 min after the start of the infusion and at 7 days. The groups of patients treated by different dose regimen were as follows: group I, 14 patients treated with t-PA 25 mg and UK 0.5 million U; group II, 20 patients given t-PA 25 mg and UK 1.0 million U; group III, 24 patients given t-PA 1.0 mg/kg and UK 0.5 million U; group IV, 33 patients treated with t-PA 1.0 mg/kg and UK 1.0 million U; and group V, 55 patients given t-PA 1.0 mg/kg and UK 2.0 million U. In groups I and II, patency of the infarct-related vessel at 90 min was only 36% and 42%, respectively. With 1 mg/kg t-PA and increasing doses of UK (groups III to V), patency ranged from 72% to 75% (overall 73%). Repeat catheterization at 7 days demonstrated reocclusion in groups III to V in 10 of 110 (9%). The patency and reocclusion rates in groups III to V were not significantly different from those in our previous study of 386 patients treated with t-PA alone (150 mg over 6 to 8 hr). In that study the patency rate of the infarct-related vessel at 90 min was 75% (p = .66) and reocclusion occurred in 15% (p = .11).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966019 TI - Reduction of atrial natriuretic factor circulating levels by endogenous sympathetic activation in hypertensive patients. AB - The effects of endogenous activation of sympathetic nervous system on systemic and regional hemodynamics and on plasma levels of atrial natriuretic factor (ANF) were studied in subjects with essential hypertension. Stimulation of sympathetic nervous system was reflex-induced by a selective deactivation of carotid baroreceptors obtained by increasing external neck-tissue pressure (NTP) by means of a neck chamber. The effects of graded levels (+30, +45, and +60 mm Hg) and one single and sustained level (+45 mm Hg for 15 min) of NTP were studied. As expected, NTP caused reflex increases in blood pressure, heart rate, and forearm vascular resistance, whereas atrial pressures did not change significantly and cardiac output tended to increase. In the studies based on graded levels of NTP, immunoreactive ANF (irANF) progressively fell (from 31.7 +/- 10 to 13.3 +/- 4 fmol/ml; p less than .05) and the changes in irANF were significantly correlated with those observed in FVR (r = -.671, p less than .001). Both hemodynamic and irANF changes were prevented by adrenergic blockade (phentolamine + propranolol). During +45 mm Hg NTP for 15 min, the levels of irANF fell both in the pulmonary artery and in the inferior vena cava. The irANF arteriovenous difference also fell during this maneuver. These data show that, in hypertensive patients, factors other than atrial wall tension may influence ANF release. They also show that endogenous sympathetic activation may reduce ANF release. PMID- 2966020 TI - A new method of inhibiting glycolysis in blood samples. AB - The maintenance of hydrogen ion concentration in blood samples at pH 5.3-5.9 immediately inhibits glycolysis. This effect is due to the inhibition of all glycolytic enzymes, as shown by measurement of various glycolytic intermediates. At the inhibitory pH at 25 degrees C, the glucose content did not decrease over a period of 8 h, but it did decrease by 1.3 +/- 1.1 mg/dl over 24 h. PMID- 2966022 TI - Population studies of Huntington's disease in Wales. AB - Long-term surveillance of Huntington's disease families living in South Wales has been undertaken since 1973. We report the updated data on prevalence and births in 101 kindreds. The trend in the births at risk of Huntington's disease has been compared with a control population in North Wales. PMID- 2966021 TI - Atrial natriuretic peptide in volume expansion-induced natriuresis in man. AB - The role of atrial natriuretic peptide (ANP) in the extracellular volume expansion (ECVE) induced natriuresis was examined in normal man under basal conditions and following dopamine blockage. Hypotonic ECVE was induced by drinking of 20 ml/kg tap water and subsequent intravenous infusion of 2 1 0, 9% saline over a period of 4 hours. This maneuver caused an increase in the plasma concentrations of ANP from 25.8 +/- 3.4 (means +/- SEM) to 59.7 +/- 6.7 fmol/ml. There was a dissociation between ANP response and urinary sodium excretion. A transient rise in glomerular filtration rate (GFR), plasma dopamine and a continuous decrease in plasma renin activity, aldosterone, vasopressin, and noradrenaline were observed. The natriuretic response to ECVE was blunted during dopamine blockade by metoclopramide, but plasma ANP, renin activity, catecholamine and vasopressin levels were not affected. However, plasma aldosterone rose. Our data are compatible with the concept that intrarenal dopamine and raised plasma concentration of ANP contribute to the natriuretic response to ECVE, but these hormonal changes do not completely explain the underlying mechanisms. PMID- 2966023 TI - Psychological aspects of amniocentesis: anxiety feelings in three different risk groups. AB - A review of the literature about parents' experiences with amniocentesis is given in the first part of this paper. In the second part the results of a follow-up study in Belgium are presented. Three groups of women who had amniocentesis performed because of advanced maternal age, a previous child with Down syndrome or a previous child with neural tube defect, respectively, were interviewed at home about their experiences. Anxiety feelings were different between groups but also showed considerable variation within each group. The overall psychological evaluation of the procedure was positive, so that the majority of the women would opt for amniocentesis in a subsequent pregnancy and would recommend it to others. Later follow-up contact by mailed questionnaire revealed that almost all women elected for their subsequent pregnancies to be monitored by amniocentesis. PMID- 2966024 TI - High affinity aldosterone binding sites (type I receptors) in rat heart. AB - 1. The use of the receptor stabilizing agent sodium molybdate, and of RU26988 to exclude [3H]-aldosterone binding from Type II glucocorticoid receptors, has enabled the characterization of high affinity Type I aldosterone binding sites in rat atrial and ventricular cytosols. 2. In adult male and female rats the affinity of binding (Kd 4 degrees C) is approximately 1-2 nmol/l for both atria and ventricles; specificity of binding is similar to that for Type I sites in classical aldosterone target tissues (aldosterone = corticosterone much greater than dexamethasone). 3. Levels of atrial Type I sites are higher than the corresponding levels in ventricle in both males and females, whereas for Type II (classical glucocorticoid) receptors the reverse is the case; levels of both Type I and Type II sites fall over the age range examined (40 days-6 months). 4. The physiological function(s) of cardiac Type I sites, and their in vivo mineralocorticoid or glucocorticoid selectivity, remain to be explored. PMID- 2966025 TI - Sonographic findings in fetuses with common chromosome abnormalities. AB - This article has detailed the common prenatal sonographic findings that have been detected in fetuses with Down syndrome, trisomy 13, trisomy 18, Turner syndrome, and triploidy. It should be noted that not all fetuses with these five chromosomal abnormalities will have sonographically detectable malformations and that some may not be easily recognizable even at birth. It is crucial that the approach to the patient with a sonographically detected fetal malformation be thorough, systematic, and expeditious. The first step is to attempt to accurately define the abnormality. Next, syndromes with which this malformation are associated should be researched and a differential diagnosis constructed. Based on this differential, accompanying findings with which this abnormality is often associated should be looked for sonographically. A detailed history including pedigree information, possible teratogen exposure, consanguinity, and ancestry is imperative. In some cases a Mendelian disorder may be identified by obtaining a thorough prenatal genetic history. In virtually all cases of prenatally detected fetal malformations, chromosome analysis is indicated. Amniocentesis is the most common means of fetal chromosome analysis, but usually takes at least 2 weeks for results. In cases where the gestation is approaching the legal limit for elective termination, rapid karyotyping by percutaneous umbilical cord sampling should be considered and referral to a center familiar with this procedure is suggested. Chromosome analysis from fetal blood takes approximately 3 days to accomplish. Alternatively, rapid karyotyping by transabdominal chorionic villus biopsy may also be accomplished and has been performed as late as 37 weeks gestation. This procedure may be especially advantageous in cases where oligohydramnios is present. If a chromosomal abnormality is identified, genetic counseling should be arranged to discuss prognosis, cause, recurrence risks and to help the parents through this difficult time. If applicable, the option of pregnancy termination should be offered. Not all parents faced with a chromosomally abnormal fetus will wish to terminate their pregnancy and others may be too late in gestation to consider this option. In these cases, discussions with other families raising similarly affected children, social workers, clergy, psychiatrists, and geneticists may be helpful to the parents. Late pregnancy prenatal cytogenetic diagnosis may be beneficial both to the parents in helping prepare them psychologically and emotionally for an adverse outcome and in guiding obstetric management.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966026 TI - Relationship of stressful housing conditions to the onset of diabetes mellitus induced by multiple, sub-diabetogenic doses of streptozotocin in mice. AB - The effect of the stress of crowded housing conditions (10 mice/cage) on the onset of diabetes after multiple, sub-diabetogenic doses of streptozotocin (MSZ) in male C57BL/KsJ mice was investigated. Prior to MSZ treatment, the group-housed and individually-housed animals had similar plasma glucose levels, while the former group's plasma corticosterone (CS) levels were elevated (54 +/- 8 ng/ml, p less than 0.03; 166% of the latter group). The group-housed animals became hyperglycemic (253 +/- 23 mg/dl) 2 days after the MSZ (day 7), with maximum hyperglycemia (506 +/- 23 mg/dl) developing by day 10. The individually housed animals did not become hyperglycemic until day 10 (303 +/- 24 mg/dl, p less than 0.001), and did not reach maximal hyperglycemia until between days 31 and 46, when plasma glucose levels were no longer different from the group-housed mice (507 +/- 37 mg/dl). There was a significant and progressive rise in CS levels of the stressed animals, reaching 218 +/- 25 ng/ml at day 46. The rise in CS of the unstressed animals was not significant until day 46, when the mean value reached 96 +/- 19 ng/ml (p less than 0.001 vs. basal). However, even at the conclusion of the experiment, the mean CS in the stressed animals was still 227% of that in the unstressed group (p less than 0.001). These studies demonstrate that the effects of stress (biochemically documented as an increase in CS levels) act synergistically with streptozotocin to promote an earlier onset of diabetes mellitus in males of this murine strain. PMID- 2966028 TI - Physical disability: the role of the physical scientist in the health service. A report of the Institute of Physical Sciences in Medicine. AB - The Report of the Royal College of Physicians on Physical Disability in 1986 and Beyond gives details of a Medical Disability Service to redress the present imbalance in the provisions for the physically disabled. An integral part of this service should be the provision of adequate scientific and technical resources, which could be established by making four or five new Medical Physics and Clinical Engineering appointments annually in each Region for the next ten years. PMID- 2966027 TI - A comparison of measurements of cerebral blood flow in the rabbit using laser Doppler spectroscopy and radionuclide labelled microspheres. AB - Laser Doppler spectroscopy has been evaluated for the measurement of cerebral blood flow (CBF) by correlation with simultaneous measurements by radionuclide labelled microspheres. The experimental procedures were carried out on five anaesthetised rabbits. The cortical tissue was exposed by means of a small burr hole and illuminated by a helium neon laser (632.8 nm). Reflected light was detected using a silicon photodiode, and CBF was calculated continuously from the power of the frequency weighted Doppler spectrum in the reflected light. Three successive measurements of CBF were made using the microsphere technique. Following an initial baseline measurement, CBF was increased by an infusion of metaraminol and then reduced by controlled haemorrhage. Laser Doppler spectroscopy provided continuous monitoring of blood flow fluctuations and during the haemorrhage it was possible to demonstrate CBF autoregulation until the mean blood pressure fell below 6.7 kPa (50 mmHg). A regression analysis was performed between the simultaneous CBF measurements from the two techniques using a least squares best fit straight line analysis (r = 0.92, P less than 0.001). It was concluded that the flow computed from laser Doppler spectroscopy varied linearly with CBF and offers the unique advantage of continuous and instantaneous measurements even during nonsteady state flow. PMID- 2966029 TI - Atrial natriuretic peptide during acute treatment of congestive heart failure. AB - Atrial natriuretic peptide (ANP) induces potent diuretic/natriuretic, vasorelaxing and aldosterone inhibitory effects. Increased plasma levels in congestive heart failure (CHF) have been reported. The aim of this study was to investigate plasma immunoreactive ANP (ir-ANP) levels during acute treatment of CHF. Seven patients with CHF underwent cardiac catheterization. Ir-ANP plasma levels were followed up to two h after administration of an orally given phosphodiesterase inhibitor (Milrinone); a substance with positive inotropic and peripheral vasodilating properties. In all patients cardiac output increased and cardiac filling pressures decreased markedly. Initially high ir-ANP plasma levels decreased. Our patients did not have an increased blood volume. It is concluded that plasma ir-ANP levels in the pulmonary artery rapidly decrease when atrial pressure is reduced. These data suggest that atrial pressure is the major determinant for release of ir-ANP in man and that the ability to respond quickly to changes in cardiac filling pressures is maintained in patients with severe CHF. Plasma ir-ANP levels may also become useful as an index of the degree of heart failure and serve as a tool in monitoring response to drug therapy. PMID- 2966030 TI - Comparison of forearm plethysmographic methods with brachial artery pulsed Doppler flowmetry in man. AB - Blood flow in the forearm was evaluated using simultaneous measurements of pulsed Doppler flowmetry and strain-gauge plethysmography in 32 normal subjects and 91 patients with sustained essential hypertension. The two determinations of blood flow were strongly correlated (r = 0.58). Measurements with strain-gauge plethysmography reflected changes in blood flow velocity but were poorly correlated with changes in arterial diameter as measured by pulsed Doppler flowmetry. The latter method permits evaluation of instantaneous variations in blood flow velocity and detection of active modifications of arterial diameter. PMID- 2966031 TI - T-cell gene rearrangements and the diagnosis of T-cell neoplasms. AB - Rearrangements of the T-cell antigen receptor genes serve as unique, clonal tumor markers of T-cell neoplasms. This approach provides a reliable and sensitive diagnostic tool to document both clonality and lineage of T-cell lymphoproliferative processes. PMID- 2966032 TI - Structure activity relationship of cholesterol and steroid hormones with respect to their effects on the Ca2+-stimulated ATPase and lipid fluidity of synaptosomal plasma membranes from dog and rabbit brain. AB - 1. Binding of liposomal-[14C]-cholesterol into dog brain synaptosomal plasma membranes (SPM) follows a sigmoid path (Hill coefficient h = 1.96 +/- 0.12). 2. Ca2+-stimulated ATPase activity increased (approx. 95%) at a concentration range (0.1-0.4 mM) of liposomal cholesterol and testosterone (up to 10 microM). While progesterone up to 10 microM decreased the enzyme activity (approx. 70%) in dog and rabbit brain SPM. 3. Fluorescence anisotropy, [(r0/r)-1]-1, of 1,6-diphenyl 1,3,5-hexatriene (DPH) was 1.04 +/- 0.04 in rabbit brain SPM and 1.72 +/- 0.09 in dog brain SPM. 4. Lipid phase separations at 23.5 +/- 1.2 degrees C in dog brain SPM and at 17.2 +/- 0.9 degrees C in rabbit brain SPM were observed. In dog brain SPM it increased to 33.4 +/- 1.8 degrees C, while in rabbit brain SPM was abolished after treatment with liposomal cholesterol. 5. The allosteric inhibition of Ca2+-stimulated ATPase by Na+ ions showed a Hill coefficient h = 1.72 +/- 0.15 and h = 1.48 +/- 0.08 for dog and rabbit brain SPM respectively, which increased to h = 2.83 +/- 0.55 and h = 2.34 +/- 0.35 after treatment with liposomal cholesterol. PMID- 2966034 TI - Viral hepatitis in dialysis units: a changing scenario. PMID- 2966033 TI - A new low-dose estrogen oral contraceptive combination: effect on endocrine parameters and lipid status. AB - In this study the variations of endocrine and metabolic parameters of 39 women treated with a new oral contraceptive combination containing 20 mcg of ethinylestradiol and 150 mcg of desogestrel were examined. Gonadotropins and androgen presented a significant decrease, showing the efficiency of suppression of the pituitary-ovarian activity and of the contraceptive effect. The significant increase of SHBG and the corresponding reduction of free testosterone reflect the estrogenic environment of this pill and the low androgenicity of desogestrel. During the observation period, plasma levels of total cholesterol, trglycerides, LDL-C and HDL2-C were not altered, whereas total phospholipids, HDL C and HDL3-C increased significantly. Besides a general positive effect on the main lipid parameters involved in atherogenic process, this pill also presented an optimal subjective tolerability. PMID- 2966035 TI - Multicomponent noninvasive physiologic monitoring of circulatory function. AB - A multicomponent noninvasive monitoring system for high-risk critically ill patients was developed and tested in a series of 58 high-risk surgical patients. The system was comprised of a thoracic electric bioimpedance (TEB) method for cardiac output estimation, laser-Doppler velocimeter for small vessel flow, transcutaneous oxygen (PtcO2) sensor for tissue oxygenation, pulse oximeter for arterial hemoglobin saturation (SaO2) and finger pressor sensor for arterial pressure. In a series of critically ill patients, this noninvasive system was compared with the widely used invasive monitoring system with systemic and pulmonary artery catheters. Despite theoretical limitations of TEB as a measure of cardiac output, there was a satisfactory correlation (r = .83) of cardiac output measured by thermodilution (TD) and TEB throughout a wide range of severe illnesses. Moreover, changes in TEB cardiac output satisfactorily tracked changes in TD cardiac output (r = .91). There was poor correlation between TD cardiac output and BP or heart rate. The incidence of abruptly changing physiologic patterns was observed with simultaneously monitored invasive and noninvasive systems. Descriptions were made of the central circulation, peripheral perfusion, and pulmonary function; common interactive responses of these circulations were also evaluated. Monitored events defined as abrupt reductions in these variables often occurred from reduced cardiac output, PtcO2 or PtcO2/PaO2 index, and infrequently from unanticipated reductions in SaO2. Recovery from the nadir of the monitored event was associated usually with improvements in flow, PtcO2, or both. Hemodynamic and oxygen transport patterns of a few commonly encountered patterns are described. PMID- 2966036 TI - Evidence for a role of endorphins in the cardiovascular pathophysiology of primate shock. AB - Using the opiate receptor antagonist naloxone, we tested the hypothesis that endorphins act on opiate receptors to cause cardiovascular depression in primate shock. Mean arterial pressure (MAP), cardiac output, and left ventricular contractility (LV dP/dtmax) were measured in 34 anesthetized cynomolgus monkeys. Hemorrhagic shock was induced by bleeding into a heparinized reservoir to achieve (t = 0) and maintain MAP at 45 mm Hg. At t = 60 min, the reservoir was clamped and the animals were treated with 2 mg/kg plus 2 mg/kg.h naloxone (n = 5) or 0.9% NaCl as a control (n = 5). There were no significant differences in the cardiovascular responses to naloxone and saline when acid-base balance and core body temperature were not controlled. Pressor responses to naloxone, however, were present in proportion to arterial pH and body temperature. When these factors were controlled, naloxone (n = 6) significantly increased MAP and LV dP/dtmax by 48% and 83%, respectively, whereas saline (n = 6) had no significant effect. Blood was reinfused at t = 120 min, and survival rate at 72 h was significantly (p = .01) higher with naloxone (3/6) than saline controls (0/6). In the endotoxic shock model, cynomolgus monkeys were treated with 2 mg/kg plus 2 mg/kg.h naloxone (n = 6) or 0.9% NaCl (n = 6) when MAP reached 75 mm Hg or its nadir 60 to 90 min after Escherichia coli endotoxin, 5 mg/kg iv. Naloxone significantly increased MAP and LV dP/dtmax by 24% and 22%, respectively, whereas saline had no effect. Survival rate at 48 h was significantly (p = .01) higher with naloxone (6/6) than saline (1/6). Plasma beta-endorphin and beta-lipotropin concentrations rose three to five-fold in both shock models and were not affected by treatment. We conclude that endorphins are activated in primate shock and act on opiate receptors to contribute to the cardiovascular depression found with hemorrhage and endotoxemia. PMID- 2966037 TI - Practical ethics in pediatrics. PMID- 2966038 TI - Reported adverse reactions to and consumption of nonsteroidal anti-inflammatory drugs in Denmark over a 17-year period. AB - Over the 17-year period 1969-1985, 2,721 reports of 3,521 suspected adverse drug reactions (ADRs) associated with non-steroidal anti-inflammatory drugs (NSAIDs) were submitted to the Danish Committee on ADRs. The results are presented together with the consumption of each drug during the same period. The total sale of NSAIDs showed a four-fold increase during the 17 years, the average corresponding to a permanent intake by 2.2% of the population. The number of reported ADRs per defined daily dose (DDD) sold was markedly lower for "older" drugs like the butazones, indomethacin, ibuprofen, naproxen, ketoprofen, and fenoprofen than for the drugs marketed during the last decade. These differences could not be accounted for by the well-known biases attached to spontaneous ADR reporting. Of 67 fatal reactions, 25 were due to bleeding or perforation of a gastric ulcer, mostly during treatment with indomethacin and naproxen, and in elderly people, and 27 were caused by bone marrow depression or leukaemia, begun mostly during treatment with butazones, but some with indomethacin and naproxen as well. It is pointed out that all reports on fatal bone marrow depression associated with butazones were submitted before 1976 and that the fact that none have been received since that time could be a result of better understanding of the proper dosage of the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966039 TI - Inspiratory muscle strength in asthma. AB - Augmentation of inspiratory muscle strength (Pimax) represents an adaptive response to airway obstruction. We explore the possibility that respiratory muscle weakness may herald hospital admission during acute bronchospasm. The Pimax measured 81 +/- 25 percent of a predicted value in 20 patients with acute bronchospasm (forced expiratory volume in one second, 36 +/- 17 percent predicted). Pimax was related to both hyperinflation (functional residual capacity, as percent predicted) and body weight (subjects were 122 +/- 29 percent ideal body weight), but not to the degree of airway obstruction per se. Furthermore, measurements of axial (craniocaudal) motion of the rib cage and asynchrony of rib cage and abdominal motions during tidal breathing did not correlate with either the degree of air flow obstruction or Pimax. We conclude that little if any respiratory muscle weakness occurs with bronchospasm. Furthermore, Pimax does not correlate with the degree of airway obstruction and does not explain abnormalities of rib cage and abdominal motion associated with asthma. PMID- 2966041 TI - Foster home recruitment: a market research approach to attracting and licensing applicants. AB - Reports differ concerning the relative recruitment productivity of the various possible resources. This report from Hennepin County, Minnesota, of a three-year recruitment study involving a large sample, with well-detailed findings, offers much to be considered by foster family care services. PMID- 2966040 TI - Recruiting adoptive parents for children with developmental disabilities. AB - One agency describes a systematic approach to recruitment of prospective adoptive families for children with developmental disabilities. The recruitment methodology, preliminary conclusions, and implications for future recruitment of families are presented. PMID- 2966043 TI - [Normal CT anatomy of the temporal bone]. PMID- 2966042 TI - [Atlanto-axial instability in Down's syndrome]. PMID- 2966044 TI - [Computed tomography in the diagnosis of intraventricular tumors]. PMID- 2966045 TI - [CT diagnosis of intracranial cholesteatoma]. PMID- 2966046 TI - [Delayed CT omnipaque myelography in 67 cases of clinically suspected syringomyelia]. PMID- 2966047 TI - [Clinico-radiologic analysis of malignant lymphoma of the colon (report of 17 cases)]. PMID- 2966048 TI - [Clinical application of transcatheter internal iliac artery embolization]. PMID- 2966049 TI - [Evaluation of pneumography combined with needle biopsy in the diagnosis of soft tissue masses (a report of 55 cases)]. PMID- 2966050 TI - [Diagnosis of pulmonary lesions by xerotomography (an analysis of 35 cases)]. PMID- 2966051 TI - [X-ray diagnosis of diastematomyelia (a report of 14 cases)]. PMID- 2966052 TI - [An experimental study of sodium morrhuate as an agent for arterial embolization]. PMID- 2966053 TI - [Autopsy of 2 siblings with Huntington's disease]. PMID- 2966054 TI - The efficacy of loop colostomy for complete fecal diversion. AB - In an attempt to evaluate the real efficacy of loop colostomy for fecal diversion, the authors studied 62 patients previously colostomized under emergency conditions. Radiologic series of the abdomen were taken after 200 gm of barium meal. The results showed that the colostomy provided complete diversion of the radiologic contrast in 53 patients (85 percent) and incomplete diversion in nine patients (15 percent). Analysis of the results revealed that incomplete fecal diversion was: 1) observed as of the 86th postoperative day, with a significantly higher frequency following the 10th postoperative month, and 2) significantly correlated with either colostomy retraction or prolapse. The authors present a diagram showing a possible interaction of factors responsible for incomplete fecal diversion in loop colostomy and conclude that: 1) retraction is probably the basic contributing factor for colostomy failure; 2) the prolapse, once reduced, propitiates sinking of the stoma, facilitating colostomy failure; 3) the common assumption that loop colostomy eventually fails to provide complete fecal diversion is further supported; 4) loop colostomy assures, over its usual duration, a satisfactory defunctionalization of the colon; and 5) use of improved techniques of colostomy construction may prolong complete fecal diversion. PMID- 2966056 TI - Malignant peritoneal mesothelioma. A clinical and laparoscopic study of ten cases. AB - A retrospective study of 10 cases of malignant peritoneal mesothelioma from two centers is reported. All cases were initially diagnosed between 1967 and 1986 utilizing laparoscopy and the histologic interpretation of laparoscopic biopsy samples. Subsequently, the original diagnosis was confirmed by two independent pathologists employing both histological and histochemical techniques. In five cases immunohistochemical studies were also performed. The clinical findings and course of the disease were similar to other reported series. Laparoscopic findings of mesothelioma were indistinguishable from metastatic peritoneal neoplasms. However, the presence of homogeneous spreading of nodules, plaques, or fleshy masses on both parietal and visceral peritoneum; the absence of direct or indirect signs of other abdominal neoplasms; and the absence of hepatic metastases or the possible presence of nodules or plaques on Glisson's capsule without any parenchymal involvement, when observed, allowed the laparoscopist to suspect the disease in four of 10 cases. Laparoscopy may be useful in detecting mesothelioma. The diagnosis is mainly morphological, but even morphology has its inherent limitations. Further studies are necessary to improve the diagnostic accuracy of this tumor. PMID- 2966055 TI - Effects of intravenous ethanol on hepatic and pancreatic blood flow in dogs. AB - Changes in hepatic and pancreatic blood flow in response to ethanol infusion were determined simultaneously and continuously in anesthetized dogs, using a transit time ultrasonic flowmeter and a laser-Doppler flowmeter. In addition, the effect of intravenous ethanol on exocrine pancreatic secretion was investigated. With a background infusion of secretin, ethanol (1.3 g/kg body wt) was infused intravenously over a 40-min period. Ethanol infusion significantly increased blood flow in the common hepatic artery (by 49%, at the time of the cessation of ethanol infusion), and this increased flow was maintained for 60 min after the cessation of ethanol infusion. In contrast, blood flow in the portal vein was not altered significantly by ethanol. Pancreatic blood flow and secretion showed no significant difference from those seen in the controls. Our data suggest that intravenous ethanol induces a redistribution of the splanchnic blood flow. The increased hepatic arterial flow seen in response to ethanol may play an important role in preventing ethanol-induced hypoxic liver damage. PMID- 2966057 TI - [Laparoscopy in fever of unknown origin]. PMID- 2966058 TI - [Reopening of long-segment occluded aortocoronary venous bypasses. Short- and long-term results]. AB - Recanalization of an occluded aortocoronary vein bypass was achieved in six out of seven patients (aged 57 +/- 4 years) 3-40 months after operation and 2-16 weeks after occlusion. The recanalization was done with a specially modified catheter introduced into the stump of the occluded vessel: urokinase was infused followed by balloon dilatation. Subsequent treatment included administration of nitrates, calcium antagonists, aspirin and heparin for at least three months. Repeat coronary angiography after 3-6 months (mean 4.3 +/- 1.0 months) demonstrated vessel patency in four of the six bypasses. One of them, severely stenosed, was again successfully dilated. It is concluded that bypass recanalization can be attempted if the occlusion occurred only a few weeks previously, angiography demonstrates a bypass stump and the state of the peripheral coronary artery makes good outflow likely. PMID- 2966059 TI - Efficacy and tolerability of an injectable formulation of tiaprofenic acid in patients with acute back and neck pain. PMID- 2966060 TI - A comparison of the insulin and insulin-like growth factor I receptors from rat brain and liver. AB - Recent studies of the human brain insulin receptor raise the possibility that some of the differences described between rat brain insulin receptors and rat liver insulin receptors may have been due to the presence of insulin-like growth factor I (IGF-I) receptors in the brain. This study details side by side comparison of the alpha-subunits from insulin and IGF-I receptors in rat brain and liver. Insulin and IGF-I receptors are present in rat liver and brain, although IGF-I receptors are at a very low concentration in rat liver. These insulin and IGF-I receptors are clearly separate entities based on relative ligand affinities, antigenicity, and apparent molecular size. The alpha-subunits of the brain receptor for both insulin and IGF-I are about 10K lower in molecular size than the corresponding alpha-subunits from liver. These brain alpha-subunits are less sensitive to neuraminidase digestion than the corresponding liver alpha subunit, although they do bind to wheat germ agglutinin. After treatment with endoglycosidase-F the deglycosylated apoproteins for both receptors from liver and brain have similar mol wt, suggesting that the observed differences were due to differences in N-linked glycosylation. The significance of differences in N linked glycosylation between the brain receptors for insulin and IGF-I and the corresponding receptors in liver is not known. PMID- 2966061 TI - Characterization of insulin-like growth factor I receptors in the median eminence of the brain and their modulation by food restriction. AB - High affinity binding sites for 125I-labeled [Thr59]insulin-like growth factor I (IGF-I) were measured in rat median eminence by in vitro autoradiography with slide-mounted sections of frozen rat brain. Specific binding of 0.1 nM iodo [Thr59]IGF-I to brain slices reached maximum by 12 h at 4 C and was unchanged at 24 h. Densitometry by computer digital image analysis of autoradiographic images indicated that specific binding of iodo-[Thr59]IGF-I to the median eminence was reversible. The specificity of binding was evaluated with competition of iodo [Thr59]IGF-I with unlabeled [Thr59]IGF-I, rat IGF-II (multiplication-stimulating activity), and porcine insulin. All were recognized by the binding site, but the rank order of potency was [Thr59]IGF-I greater than IGF-II greater than insulin. Somatostatin was completely ineffective. Further, an antibody against the rat IGF II receptor did not block binding of iodo-[Thr59]IGF-I to the median eminence. Fourteen days of food restriction (75% of food intake of controls) resulted in significant weight loss and reduction of plasma immunoreactive IGF-I in six food restricted rats (0.9 +/- 0.1 U/ml) compared with values in six controls (2.6 +/- 0.5 U/ml; P less than 0.001). Binding of 125I-labeled [Thr59]IGF-I in the median eminence was significantly increased in the food-restricted rats, primarily due to an increase in the concentration of iodo-[Thr59]IGF-I-binding sites in the median eminence; the affinity (Kd) of binding was unchanged. The results indicate that the median eminence has type I IGF-I receptors, which become more numerous under metabolic conditions associated with decreased caloric intake and lowered plasma IGF-I levels. PMID- 2966062 TI - Receptor-mediated endocytosis and lysosomal processing of insulin-like growth factor I by mitogenically responsive cells. AB - The processing of insulin-like growth factor I (IGF-I) by MG-63, an IGF-I responsive human osteosarcoma cell line, has been investigated. At 37 C, the binding of [125I] IGF-I to monolayers of MG-63 cells reaches a maximum after about 1 h and slowly declines thereafter. The addition of the lysosomotropic agents methylamine, chloroquine, and monensin to the binding medium prevents the decline in [125I]IGF-I binding observed in the untreated cells and causes a 1.5- to 3-fold increase in cell-associated radioactivity after 4 h. Leupeptin, an inhibitor of lysosomal proteases, and colchicine, an inhibitor of endosomal transport, also increase cell-associated [125I]IGF-I. Three observations indicate that the increased radioactivity associated with the treated cells is the result of intracellular accumulation of the ligands and not the result of an increase in cell surface IGF binding. First, no increase in [125I]IGF-I binding is observed in cells preincubated with methylamine at 37 C but transferred to 4 C (where endocytosis is inhibited) before the addition of the radiolabeled ligands. Second, the increased radioactivity bound by methylamine-treated cells is not removed by washing the cells with dilute acid, a treatment that removes surface bound [125I]IGF-I. Third, in leupeptin-treated cells [125I]IGF-I accumulates in a subcellular fraction with properties characteristic of lysosomes. Both alpha IR-3 (100 nM), an antibody that specifically inhibits binding to the type I IGF receptor, and high concentrations of insulin (900 nM) inhibit the accumulation of [125I]IGF-I by methylamine-treated cells, indicating that internalization of IGF I occurs through the type I IGF receptor and not through the type II IGF receptor or the IGF-binding protein(s) that is also present on these cells. These results demonstrate that in MG-63 cells IGF-I is endocytosed via the type I IGF receptor and that the endocytosed hormone is degraded, at least in part, in lysosomes. These findings are similar to those described for the processing of insulin and other growth factors by their target cells and extend further the homology between IGF-I and these other agents. PMID- 2966063 TI - Stimulation of growth hormone secretion by central administration of atrial natriuretic polypeptide in the rat. AB - Intracerebroventricular (icv) injection of alpha-human atrial natriuretic polypeptide (alpha hANP) or alpha-rat ANP (0.6 and 3 nmol/rat) elicited an increase in plasma GH levels both in conscious freely moving rats and in urethane anesthetized rats when given at the trough of spontaneous GH secretion. Antiserum specific for rat GRF did not affect the plasma GH increase induced by icv injection of alpha hANP. Intracerebroventricular injection of alpha ANP (3 nmol/rat) failed to stimulate GH secretion in conscious rats pretreated with cysteamine (30 mg/100 g BW, sc), a depletor of somatostatin (SRIF), and in conscious rats during constant iv infusion of SRIF (55 ng/ml). GH release induced by iv injection of synthetic rat GRF (200 ng/100 g BW) was exaggerated by alpha hANP (3 nmol/rat, icv) in conscious rats. These results suggest that central ANP stimulates pituitary GH secretion possibly by inhibiting SRIF release from the hypothalamus in the rat. PMID- 2966064 TI - Sodium excretion and atrial natriuretic peptide levels during mineralocorticoid administration. A mechanism for the escape from hyperaldosteronism. AB - Urinary sodium excretion initially decreases when mineralocorticoid levels are increased, but if high plasma levels of hormone are maintained, sodium excretion rises to again equal sodium intake. To ascertain if atrial natriuretic peptide (ANP) plays a role in reestablishing sodium balance during mineralocorticoid ingestion, 0.3 to 0.5 mg per day of fludrocortisone were administered for 18 days to four healthy male subjects. The average daily intake of sodium was regulated at 180 +/- 2 meq. ANP levels rose from a mean of 91.7 +/- 13.0 pg/ml during the control week to 179.7 +/- 39.2 pg/ml during the final week on fludrocortisone (p less than 0.05). Urinary sodium excretion fell 27% immediately after fludrocortisone administration was initiated but returned to baseline levels in an average of 5 days. Levels of ANP, normalized for each subject to the mean of his control week values, correlated with the amount of sodium excreted in the subsequent 24 hours (p less than 0.05). Simultaneous with the rise in ANP values, levels of plasma renin activity (PRA) and aldosterone decreased. ANP concentrations throughout the study were inversely correlated with PRA and aldosterone levels (p less than 0.001 for both correlations). Values of serum osmolality and plasma arginine vasopressin did not change significantly during the study. The results obtained demonstrate that increased secretion of ANP is associated with escape from the sodium retaining effect of chronically high mineralocorticoid levels in man and suggest that ANP plays a prominent role in the mechanism of this escape. PMID- 2966065 TI - Activation of the receptor kinase domain of the trk oncogene by recombination with two different cellular sequences. AB - A new chimeric oncogene, trk-2h, has been generated by recombination of two segments of MDA-MB231 human breast carcinoma cell line DNA after transfection in NIH/3T3 cells. The rearranged DNA segments form a fused transcriptional unit. Sequences at the 3' end are homologous to the tyrosine kinase receptor moiety found in the trk oncogene which resembles a truncated growth factor receptor lacking part of its extracellular domain (Martin-Zanca et al., 1986). The 5' sequence of the trk-2h oncogene is contributed by a gene which is expressed in all human cells tested, and is not related to any known gene. Transfection of the receptor kinase domain DNA fragment into NIH/3T3 cells generated another oncogene, trk-3mh, which contains a mouse-specific sequence fused 5' to the receptor kinase. All three trk recombinants have the receptor kinase moiety fused to an activating amino terminus at the same nucleotide in their transcriptional product. PMID- 2966067 TI - Tryptic cleavage inhibits but does not uncouple Ca2+ATPase of sarcoplasmic reticulum. AB - Sarcoplasmic reticulum Ca2+ATPase is cleaved by trypsin at two sites, T1 and T2. Cleavage at T1 is complete, whereas only about 50% of the Ca2+ATPase is digested at the T2 site. In the absence of Ca2+ ionophor, Ca2+-ATPase activity of the digested enzyme remains virtually unchanged. In the presence of Ca2+ ionophor, however, the calculated specific activity of the doubly cleaved Ca2+ATPase is decreased by about 40%. The decrease in Ca2+ transport activity is much more rapid than cleavage of the T2 site, and could be correlated with an increased leak of Ca2+ from the digested vesicles. We obtained evidence that this leakiness is independent of the digestion of the Ca2+ATPase itself and is presumably due to the digestion of some other components of the sarcoplasmic reticulum vesicles. Examination of steady-state phosphoenzyme levels resulting from phosphorylation by ATP and Pi, or dephosphorylation by ADP or ADP/EGTA revealed no difference between the digested and the undigested Ca2+ATPase indicating no change in the equilibria caused by the T2 cleavage. Analysis of the substrate concentration dependence of the Ca2+ATPase activity also led to the conclusion that the digestion at T2 reduced the Vmax of ATP hydrolysis but leaves the Km unchanged. The above results are consistent with the model that cleavage at the T2 site reduces the turnover rate of the Ca2+ATPase reaction cycle by about 40% by slowing down or altering the rate-limiting step without affecting the equilibrium constants of the examined steps. We found no evidence of true uncoupling of Ca2+ transport from ATP hydrolysis correlated with cleavage at the T2 site. PMID- 2966066 TI - Three-dimensional reconstruction of maltoporin from electron microscopy and image processing. AB - Two dimensional crystals of maltoporin (or phage lambda receptor) were obtained by reconstitution of purified maltoporin trimers and Escherichia coli phospholipids by detergent dialysis. Two different trimer packing forms were observed. One was hexagonal (a = 7.8 nm) and one rectangular (a = 7.8 nm, b = 13.6 nm). In this paper we describe the three-dimensional structure of maltoporin, deduced from the study of the rectangular form by electron microscopy and image processing. At a resolution of approximately 2.5 nm, maltoporin trimers form aqueous channel triplets which appear to merge into a single outlet at the periplasmic surface of the outer membrane. The pore defined by maltoporin has a similar structure to that outlined by the matrix protein. From the results of functional studies by conductance measurement, it is concluded that the three channels defined by maltoporin act, contrary to those formed by the porin (OmpF protein), as a single conducting unit. A tentative outline of the maltoporin promoter is given. Maltoporin appears to be constituted by three different domains: a major rod-like domain spanning the membrane, a minor domain located near the periplasmic surface of the membrane and finally a central domain responsible for the splitting of the channel. PMID- 2966068 TI - Plasma concentrations of atrial natriuretic peptide in patients with artificial and transplanted hearts. AB - Plasma atrial natriuretic peptide (ANP) concentrations were measured by radioimmunoassay 1 to 12 days after operation in six patients with artificial hearts. The mean levels were significantly higher in artificial heart recipients (58.2 pg.ml-1) than in controls (12.6 pg.ml-1). The mechanism underlying the raised values is not clear, although it is suggested that elevated atrial pressure may have been the stimulus for higher ANP release. In 3 patients plasma ANP concentrations were also measured 1 to 5 days after orthotopic transplantation. In all of them ANP concentrations had increased by 20 to 74% despite lower right atrial pressure. An increase in atrial tissue mass may have contributed to the raised plasma ANP after orthotopic transplantation. It also suggests that the functioning sympathetic nervous system is not a necessary condition for ANP release. PMID- 2966069 TI - Ganglioside-dependent adhesion events of human neuroblastoma cells regulated by the RGDS-dependent fibronectin receptor and proteoglycans. AB - Human neuroblastoma cells (Platt and La-N1) adhere and extend neurites on a ganglioside GM1-binding substratum provided by cholera toxin B (CTB). These adhesive responses, similar to those on plasma fibronectin (pFN), require the mediation of one or more cell-surface proteins [G. Mugnai and L. A. Culp (1987) Exp. Cell Res. 169, 328]. The involvement of two pFN receptor molecules in ganglioside GM1-mediated responses on CTB have now been tested. In order to test the role of cellular FN binding to its glycoprotein receptor integrin, a soluble peptide containing the Arg-Gly-Asp-Ser (RGDS) sequence was added to the medium. It did not inhibit attachment on CTB but completely inhibited formation of neurites; in contrast, the RGDS peptide minimally inhibited attachment or neurite formation on pFN. Once formed, neurites on CTB became resistant to the peptide. In order to test the role of cell-surface heparan sulfate proteoglycan (HS-PG), two approaches were used. First, the HS-binding protein platelet factor-4 (PF4) was used to dilute CTB or pFN on the substratum or, alternatively, added to the medium. Diluting the substratum ligand with PF4 had no effects on attachment on either CTB or pFN. However, neurite formation on CTB was readily inhibited and on pFN partially inhibited; the effects of PF4 were far greater than a similar dilution with nonbinding albumin. When PF4 was added to the medium of cells, attachment on either substratum was unaffected as was neurite outgrowth on pFN, revealing differences in PF4's inhibition as the substratum-bound or medium-borne component. In contrast, PF4 in the medium at low concentrations (1 microgram/ml) was highly inhibitory for neurite formation on CTB. The second approach utilized the addition of bovine cartilage dermatan sulfate proteoglycan (DS-PG), shown to bind to pFN as well as to substratum-bound CTB by ELISA, or cartilage chondroitin sulfate/keratan sulfate proteoglycan (CS/KS-PG) to the substratum or to the medium. At low concentrations, DS-PG but not CS/KS-PG actually stimulated neurite formation on CTB while at higher concentrations DS-PG completely inhibited attachment and neurite formation. While DS-PG partially inhibited attachment on pFN, it had no effect on neurite formation of the attached cells. Neuroblastoma cells adhered to some extent to substrata coated only with DS-PG, indicating "receptors" for PGs that permit stable interaction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966070 TI - The effect of recombinant murine interferon-gamma on the hematopoietic and immunological parameters of mice bearing metastatic Lewis lung carcinoma tumors. AB - The in vitro and in vivo effects of murine recombinant interferon-gamma (rIFN gamma) on hematopoietic and immune parameters of normal mice and of mice bearing metastatic variant Lewis lung carcinoma (LLC-C3) tumors were assessed. The in vitro addition of rIFN-gamma to bone marrow or spleen cells from normal and LLC C3-bearing mice reduced their capacity to grow into colonies in soft agar (CFU) and minimized their immune suppressive activities. In vivo studies showed that when LLC-C3 tumor-bearing mice were injected with rIFN-gamma for 2 days prior to sacrifice, there was a reduction in femoral bone marrow cellularity, CFU, and suppressor cell activity. In contrast, spleen cells of tumor-bearing mice that were injected with rIFN-gamma showed reduced blastogenesis, and increased spleen cellularity, CFU, and suppressor cell activity. Thus, short-term rIFN-gamma treatment of LLC-C3-bearing mice may be beneficial with regard to the bone marrow because it caused a decrease in hematopoiesis and suppressor cell activity, whereas it may be detrimental in the spleen because it appeared to stimulate hematopoiesis and increase splenic suppressor cell activity. The dichotomy between the in vitro versus in vivo effects of rIFN-gamma on splenic hematopoiesis and suppressor activity may be due to the stimulation of production of colony-stimulating factor (CSF) activities by spleen cells of rIFN-gamma treated mice. Our results suggest that the tumor stimulation of hematopoiesis and its associated appearance of immune suppressor cells can be both positively and negatively altered by rIFN-gamma. PMID- 2966071 TI - Occurrence of platelet basic protein, a precursor of low affinity platelet factor 4 and beta-thromboglobulin, in human platelets and megakaryocytes. AB - beta-thromboglobulin antigen, a platelet-specific secreted protein, occurs in three forms: platelet basic protein, low affinity platelet factor 4, and beta thromboglobulin. The combined level of beta-thromboglobulin antigen in megakaryocytes, measured by radioimmunoassay, was 13 +/- 7 micrograms/10(6) cells (SD, n = 6). The relative proportions of the three forms of beta-thromboglobulin antigen present within platelets and megakaryocytes were determined in cells lysed with trichloroacetic acid to minimize artifactual proteolysis. Samples were analyzed by isoelectric focusing in polyacrylamide gel with quantitative immunological detection on a nitrocellulose transfer of the gel. In platelets, the major species found was low affinity platelet factor 4 with precursor platelet basic protein as only 25% +/- 11% (SD, n = 16) of total beta thromboglobulin antigen. In megakaryocytes partially purified both from normal bone marrow aspirates and from whole marrow specimens obtained after surgery, platelet basic protein was a higher proportion of beta-thromboglobulin antigen (49% +/- 13% SD, n = 11) than was the case in platelets. beta-thromboglobulin itself was never detected under the conditions of cell lysis used. Our results suggest that platelet basic protein is synthesized in megakaryocytes and that its cleavage is associated with an earlier stage of cell development than simply maturation to platelets. Further support for the precursor status of platelet basic protein was found in the expression of predominantly this antigenic form in a human erythroleukemia cell line. PMID- 2966072 TI - Altered immunological reactivity in alveolar macrophages from patients with sarcoidosis. AB - Lung macrophages may play an important role in the pathogenesis of pulmonary sarcoidosis. In this study, the ability of pulmonary macrophages and blood monocytes from sarcoidosis patients, normal controls and disease controls to provide the accessory signal necessary for the concanavalin A-induced activation of normal blood T cells was examined. Blood monocytes from all groups supplied a significantly greater accessory signal than lung macrophages. The accessory capacity of lavage macrophages from sarcoidosis patients varied over a wide range and correlations were sought between these values and other parameters of disease activity. Whilst there was no correlation with clinical parameters, accessory function of alveolar macrophages correlated significantly with the percentage of T helper cells in bronchoalveolar lavage (BAL) fluid (p less than 0.05) and, more closely, with the T helper:T suppressor ratio in BAL fluid (p less than 0.01). This interrelationship between macrophage activity and the T cell infiltrate favours the probability that both cell types participate in the sarcoid disease process and raises the possibility that T cells of both helper and suppressor phenotypes contribute to the pathogenesis. PMID- 2966073 TI - [Effect of nicotinic acid, nicotinamide and its combination with ziksorin and phenobarbital on the UDP glucuronyl transferase activity of the hepatic endoplasmic reticulum in the rat]. AB - Nicotinic acid and nicotinamide (100 mg/kg) increase the activity of the rat liver microsomal uridine diphosphate-glucuronyltransferase by 55 and 73.8%. Administration of nicotinamide in combination with ziksorin or phenobarbital enhanced the enzyme-inducing effects of the latter. PMID- 2966075 TI - Evidence that stimulation of gluconeogenesis by fatty acid is mediated through thermodynamic mechanisms. AB - We have studied the stimulatory effects of palmitate on the rate of glucose synthesis from lactate in isolated hepatocytes. Control of the metabolic flow was achieved by modulating the activity of enolase using graded concentrations of fluoride. Unexpectedly, palmitate stimulated gluconeogenesis even when enolase was rate-limiting. This stimulation was also observed when the activities of phosphoenolpyruvate carboxykinase and aspartate aminotransferase were modulated using graded concentrations of quinolinate and aminooxyacetate, respectively. Linear force-flow relationships were found between the rate of gluconeogenesis and indicators of cellular energy status (i.e. mitochondrial membrane and redox potentials and cellular phosphorylation potential). These findings suggest that the fatty acid stimulation of glucose synthesis is in part mediated through thermodynamic mechanisms. PMID- 2966074 TI - Site directed mutants of human interleukin-1 alpha: a 1H-NMR and receptor binding study. AB - Mutant human interleukin-1 alpha proteins were constructed by oligonucleotide directed mutagenesis. Six different mutants were tested for receptor binding activity and showed no alteration with respect to the wild-type protein. Analysis of these mutants by nuclear magnetic resonance spectroscopy confirmed the structural integrity of the mutant proteins and permitted the sequence specific assignment of the histidine and tryptophan residues. PMID- 2966077 TI - Secretion and biosynthesis of atrial natriuretic factor by cultured adrenal chromaffin cells. AB - In our previous work, the existence of the precursor and mature forms of atrial natriuretic factor (ANF) within the bovine chromaffin granules has been reported. To confirm the endogenous character of these peptides, we demonstrate that nicotinic activation and depolarization by KCl increase their co-secretion from cultured chromaffin cells. The increase of intracellular levels of these atrial peptides by phorbol ester is potentiated by addition of forskolin. The release of ANF and their de novo synthesis within the cultured chromaffin cells emphasize the usefulness of this model in the study of the mechanisms of release and storage of these peptides in the neuronal tissues. PMID- 2966076 TI - Localization of the pro-sequence within the total deduced primary structure of human beta-hexosaminidase B. AB - The beta subunit of beta-hexosaminidase (beta-N-acetylhexosaminidase, EC 3.2.1.52) is synthesized in the rough endoplasmic reticulum as a prepropolypeptide. After the loss of the signal peptide and formation of an enzymatically active dimer, the pro-enzyme is either secreted from the cell or transported into the lysosome for processing to its mature form. In order to characterize the early posttranslational events we have purified nearly 1 mg of pro-hexosaminidase B from the NH4Cl containing medium of fibroblasts derived from a patient with the infantile form of Tay-Sachs disease. The partial N-terminal sequence was mapped to a position 42 residues C-terminal to the first in-frame ATG (Met residue) and 79 residues N-terminal to the known mature N-terminus. This position corresponds to that predicted for the cleavage of a 17 amino acid signal peptide generated through the use of the third rather than the first in-frame ATG as the initiation site for protein synthesis. PMID- 2966078 TI - Post-translational modification of the beta-subunit of the human fibronectin receptor. AB - Monoclonal antibody DH12, directed against the beta-subunit of the fibronectin receptor recognizes a doublet of proteins (100 and 110 kDa) in Western blots of solubilized whole fibroblasts. Pulse-chase experiments with [35S]methionine in human skin fibroblasts suggested that the two proteins might be metabolically related as precursor (100 kDa) and product (110 kDa). Endo H digestion and [3H]fucose labeling suggested that maturation converted the high-mannose oligosaccharides (100 kDa) to the endoglycosidase H resistant complex type (110 kDa). This was supported by N-glycanase digestion and by chemical deglycosylation which showed a single polypeptide. Surface iodination of intact cells labeled only the presumed mature beta-subunit. PMID- 2966079 TI - The long-term consequences of prenatal and perinatal infections. Subnormality. PMID- 2966080 TI - Principles of development of model health care programmes. Report on a WHO meeting. PMID- 2966081 TI - Changes of enzyme activity in the rat liver at different age in circadian cycle. AB - Most of the biological processes in the living organisms of both animals and man are known to be of rhythmical nature. Variability of enzymatic activity in circadian cycle depends on many factors among other on age, sexual maturity, diet as well as medication. The results obtained in our studies indicate, that the activity changes of acid phosphatase and ATP-ase Mg++ dependent in the liver of all the examined age groups were of 24 hour circadian rythm. As to the acid phosphatase activity the results of this experiments showed that in circadian cycle in all examined age groups there is only one peak of elevated activity. ATP ase Mg++ dependent showed two activity peaks appearing at the same hour both in 30 and 60 days old animals. It should be noticed that the activities of ATP-ase Mg++ dependent in 100 day old animals were two times higher than in 30 and 60 days old rats. PMID- 2966082 TI - Studies on the structure of the boundary tissue of the white rat seminiferous tubules. AB - The studies on boundary tissue of the white rat seminiferous tubules with light and electron microscopy were carried out. The wall of the tubules consists of four layers: two cellular and two amorphous ones. In cellular external sheath the characteristic intercellular fissures a network of hexagonal meshes were seen resembling the honey-combs. PMID- 2966083 TI - Analysis of deoxynivalenol as its trifluoroacetyl ester by gas chromatography electron ionization mass spectrometry. AB - Mass spectra of the trifluoroacetyl esters of the trichothecenes deoxynivalenol, diacetoxyscirpenol and T-2 toxin, as well as zearalenone, were obtained by gas chromatography-electron ionization mass spectrometry. Quantitation of deoxynivalenol as its trifluoroacetyl ester by selected ion monitoring was then possible down to 10 pg in the mass spectrometer (equivalent to 1 ng/g in a sample). The method was tested by extracting and analysing mouldy wheat which had previously given a positive result for deoxynivalenol when analysed by thin layer chromatography. The method was also suitable for the analysis of diacetoxyscirpenol and T-2 toxin; however, quantitation of zearalenone was unsuccessful because of the instability of the derivative. PMID- 2966085 TI - Composition changes in hepatic microsomal cytochrome P-450 during onset of streptozocin-induced diabetes and during insulin treatment. AB - Monospecific polyclonal antibodies to five constitutive hepatic microsomal cytochromes P-450 were prepared. These antibodies were used to monitor alterations in the content of the enzymes in livers of diabetic male rats. Within 3 wk of onset of streptozocin-induced diabetes, immunodetectable levels of RLM3 and RLM5 were decreased by 85 and greater than 95%, respectively. Insulin treatment for 1 wk reversed the decline in these isozymes and restored RLM3 to 60% and RLM5 to 53% of levels found in the untreated rat. After a 2nd wk of therapy, these levels were returned to 86 and 92%, respectively. In contrast, the levels of RLM5b and RLM6 were elevated in diabetes 1.7- and 8-fold, respectively. Insulin treatment for 1 wk only slightly decreased the levels of RLM5b but completely reduced RLM6 levels to those seen in age-matched untreated rats. After the 2nd wk of insulin treatment, the level of RLM5b was almost completely restored to normal, with no additional change in the RLM6 level. The level of a fifth enzyme, RLM5a, was not markedly altered by diabetes or by insulin treatment. The results suggest there are at least three types of responses by constituents of the cytochrome P-450 population to diabetes: no change in the microsomal content, a rapid increase when insulin level declines and restoration when insulin is supplied, and a rapid decline when insulin level declines and a restoration by insulin treatment. PMID- 2966084 TI - Insulin-like growth factor I (IGF-I) receptors on Sertoli cells from immature rats and age-dependent testicular binding of IGF-I and insulin. AB - Insulin-like growth factor I (IGF-I) binding to cultured Sertoli cells from immature rats was quantitatively evaluated. The binding of 125I-IGF-I to the Sertoli cells was specific, time- and pH-dependent and reversible. Scatchard analysis yielded a Kd of 3.5 X 10(-9) M and a binding capacity of 2080 fmol/mg protein. Competition with IGF-I resulted in a half-maximal displacement by 2 nM IGF-I, whereas insulin up to a concentration of 100 nM gave virtually no displacement of IGF-I binding. Similarly, the gonadotropic hormones follitropin and lutropin did not compete with 125I-IGF-I binding. In previous studies, it was shown that cultured Sertoli cells from immature rats bind insulin with a Kd of 1.8 X 10(-9) M and a binding capacity of 8.5 fmol/mg protein. The binding of IGF I and insulin to a total testis membrane fraction was studied using testes from immature and adult rats. In testis from 21-day-old rats, the maximal specific binding was relatively high for IGF-I (871 +/- 50 fmol/g wet weight) and relatively low for insulin (118 +/- 11 fmol/g wet weight). In adult testis, the maximal specific binding of IGF-I was 324 +/- 40 fmol/g wet weight and that of insulin was 330 +/- 17 fmol/g wet weight. The binding of IGF-I and insulin expressed as fmol bound per testis was increased 6-fold and 45-fold, respectively, between the age of 21 days and adult age. It is discussed that the numbers of receptors for IGF-I and insulin in testis may be developmentally regulated, and that IGF-I may be more important than insulin with respect to testis development and Sertoli cell maturation in the immature rat. PMID- 2966086 TI - Developmental regulation of insulin and type I insulin-like growth factor receptors and absence of type II receptors in chicken embryo tissues. AB - Chicken embryos are a suitable model for studying the role of insulin, insulin like growth factors I and II (IGF-I and IGF-II), and their receptors in embryogenesis. We show that plasma membranes from heart, liver, and limb buds, as reported earlier for brain, each have a distinct developmental profile for insulin receptors and type I IGF receptors. In heart and limb buds, IGF binding is higher than insulin binding, but in liver, insulin receptors dominate. Expression of these receptors is, therefore, developmentally regulated and tissue specific. The wide distribution of high-affinity receptors capable of mediating insulin and IGF actions in early organogenesis further supports the possible importance of this family of peptides for differentiation and growth in vertebrates. In all chicken embryo tissues studied, both IGF-I and IGF-II appeared to bind to a type I IGF receptor. We have not detected a receptor with the peptide binding and structural characteristics of the mammalian type II IGF receptor. The type II receptor was absent in embryos, liver from newly hatched chicks, and adipocytes from older chicks, which suggests that the chicken may lack this subtype of IGF receptor. PMID- 2966087 TI - T cell and OKM1-positive monocyte populations in the intestinal lamina propria mucosae and in the peripheral venous blood in Crohn's disease. AB - Nineteen patients with Crohn's disease were examined prior to operation. Peripherally circulating mononuclear cells were examined for the presence of the cell surface markers recognized by monoclonal antibodies OKT3, OKT4, OKT8 and OKM1. After surgical treatment samples of the mucosal lesions were obtained from the resected bowel segments of these patients. The inflammatory infiltrate in the lamina propria mucosae was examined immunohistologically for the presence and distribution of the cell surface markers recognized by Dako T11, OKT4, OKT8 and OKM1. In comparison to normal controls, the patients with Crohn's disease had a relative peripheral lymphocytopenia with a relative monocytosis. The intestines afflicted by active Crohn's disease showed a significant increase in macrophages and T cells in the lamina propria mucosae; both T helper and T suppressor cell populations had equally increased without a significant change in the T helper/suppressor ratio. These findings suggest a change in the equilibrium state between intravascular and intestinal mucosal lymphocytes and monocytes in the active phase of the disease. The new equilibrium can be characterized as an intestinal lymphomonocytosis with a relative lymphocytopenia and monocytosis in the peripheral blood. PMID- 2966089 TI - [Leuko- and erythrocytic deficiency of the enzyme glucose-6-phosphate dehydrogenase in populations with a high incidence of this anomaly]. PMID- 2966088 TI - Prospective study of chemotherapy in patients with metastatic gastrinoma. AB - Ten consecutive patients with metastatic gastrinoma that increased in size over time were studied prospectively during treatment with monthly cycles of streptozotocin (3 g/m2), 5-fluorouracil (1.2 g/m2), and adriamycin (40 mg/m2) to determine the response rate and time-courses of changes during chemotherapy and to assess various methods of evaluating the effect of chemotherapy. Forty percent of patients demonstrated an initial objective response (greater than or equal to 25% decrease in tumor size with no new lesions) and 60% failed chemotherapy (greater than or equal to 25% increase in tumor size or appearance of new lesions). The mean dose of streptozotocin was 27 g/m2 with objective responses occurring at 3.7 +/- 0.7 mo and failures at 4.5 +/- 0.7 mo. Responses lasted 9.7 +/- 2.8 cycles and no complete responses occurred. Survival was not significantly different in responders versus nonresponders (26 +/- 11 vs. 15 +/- 4.8 mo, p greater than 0.1). Changes in serum gastrin concentration, basal acid output, or sensitivity to a given dose of histamine H2-receptor antagonist did not reflect changes in tumor size. Computed tomography and angiography were the best methods to assess changes in tumor size during chemotherapy, whereas liver-spleen scan and ultrasound were relatively insensitive. All patients developed side effects with chemotherapy: 100% had vomiting, 80% alopecia, 40% transient proteinuria, and 20% leukopenia. The present results indicate that chemotherapy with streptozotocin, 5-fluorouracil, and adriamycin is much less effective in patients with extensive metastatic gastrinoma than previously reported. Computed tomography scanning is the method of choice to assess changes in tumor size. Changes in serum gastrin concentration, acid secretion, or tumor size assessed by liver-spleen scan or ultrasound are not sensitive indicators of the tumor response during chemotherapy. PMID- 2966090 TI - [The role of the chalone-antichalone system in hematology]. PMID- 2966091 TI - Detection of new MHC mutations in mice by skin grafting, tumor transplantation and monoclonal antibodies: a comparison. AB - Two mechanisms of major histocompatibility complex (MHC) mutations have been described in mice; gene conversion and homologous but unequal recombination. However, our knowledge of mutations in MHC is incomplete because studies have been limited almost exclusively to two haplotypes, H-2b and H-2d, while hundreds of haplotypes exist in nature; it has been biased by the use of only one procedure of screening for mutation, skin grafting. We used three procedures to screen for MHC mutations: (1) conventional techniques of skin grafting, (2) syngeneic tumor transplantation and (3) typing with monoclonal anti-MHC antibodies (mAbs) and complement. The faster technique of tumor transplantation detected mutants similar to those discovered by skin grafting technique. Screening with mAbs allowed us to detect both mutants that are capable of rejecting standard skin grafts and those that are silent in skin grafting tests, and which therefore resulted in a higher apparent mutation frequency. Two mutants of the H-2a haplotype were found that carry concomitant class I and class II antigenic alterations. Both MHC mutants silent in skin grafting tests and mutants carrying concomitant class I and class II alterations have never been studied before and are expected to reveal new mechanisms of generating MHC mutations. 1 Ethyl-1-nitrosourea (ENU) failed to induce de novo MHC mutations in male mice in our skin grafting series. PMID- 2966092 TI - [Heritability of fertility in human populations and the structure of Crow's index]. AB - Heritability of fertility was determined in populations with natural character of reproduction (Middle Asia Republics) and in populations with family planning (Middle Russia). The material was collected by interviewing the married women of postreproductive age. The heritability of fertility was estimated as a doubled coefficient of correlations between sisters for effective fertility. In populations with natural character of reproduction the heritability was about 50% and it did not differ from O in the populations with family planning. The values of heritability were used in the components of the Crow index analysis. The main contribution into broad-sense heritability of fertility comes from additive component, as shown in an Asia population (Turkmen). The values of heritability of fecundability (86%) and of postpartum sterility (66%) have been calculated for the same population. PMID- 2966093 TI - Identification of the DNA binding domain of the phage lambda cII transcriptional activator and the direct correlation of cII protein stability with its oligomeric forms. AB - The bacteriophage lambda transcriptional activator protein cII is a DNA-binding protein that coordinately regulates transcription from phage promoters important for lysogenic growth. We have genetically and structurally characterized more than 80 different single amino acid substitutions in this 97-amino-acid protein. A subset of 25 of these variant proteins was utilized for detailed biochemical analysis, which allows us to define specific domains critical for sequence selective DNA recognition, nonspecific DNA binding, and protein oligomerization. The mutation studies also demonstrated the remarkable correlation of oligomeric structure of cII protein to its stability within the bacterial host. An Escherichia coli HtpR- strain has been identified that greatly stabilizes these highly unstable cII mutants. PMID- 2966095 TI - [Percutaneous transluminal coronary angioplasty (PTCA). Indications and results- status after 10 years' experience]. PMID- 2966096 TI - [Monoaminergic regulation of the levels of beta-endorphin-like immunoreactivity (beta-ENDi) in rat hypothalamic nuclei]. PMID- 2966094 TI - Down's syndrome: a pathology involving the lack of balance of reactive oxygen species. AB - Mongolism, or Down's syndrome (DS) is an inborn pathology due to trisomy 21. Apart from mental deficiency and anatomical abnormalities, premature aging is characteristic for this disease. This paper mainly reviews data on the generation of reactive oxygen species (ROS) and antioxidative defense mechanisms in DS, and on the sensitivity of DS cells to model sources of ROS. Changes in the antioxidative defense of DS cells are due mainly to the location of the gene for (Cu,Zn)-superoxide dismutase on chromosome 21. It is suggested that a disturbance of balance of ROS may be a key point in the pathogenesis of DS. PMID- 2966097 TI - Atrial natriuretic peptide in infants and children. AB - Atrial natriuretic peptide (ANP) was measured in the plasma of 192 normal infants and children aged 1 day to 18 years. Plasma ANP was high during postnatal adaptation, particularly in premature infants. In 96 infants and children aged 4 months to 18 years, plasma ANP was similar to values obtained in 7 healthy adult volunteers (23.9 +/- 11.9 vs. 25.7 +/- 4.6 fmol/ml). There was no significant relationship between ANP and age. ANP is elevated about twofold in full-term neonates being 3-4 days of age, and returned to normal thereafter. It is concluded that ANP is raised during the postnatal adaptation. This hormone is possibly involved in the postnatal volume contraction and may antagonize vasoconstrictor hormones that are elevated during the postnatal period. PMID- 2966098 TI - PGM2 *2 (Atkinson) exists in Germany. PMID- 2966099 TI - Controversy concerning paternal age effect in 47, + 21 Down's syndrome. PMID- 2966100 TI - Hyperreactivity of adult BALB/c mice tolerized at birth with TNP-ovalbumin. AB - BALB/c mice injected with 0.2 mg TNP-ovalbumin (OA) within 24 hours after birth showed reduced levels of functionally active TNP-specific B cells and, accordingly, of plaque-forming cells (PFC) after challenge with carrier-bound TNP until the age of 8 weeks. Yet, when B cells of tolerized mice were cultured in the absence of antigen, a significant number of anti-TNP PFC were detected. Challenge of neonatally tolerized mice at older age with T-dependent or T independent antigens led to a continuously increasing response towards TNP, which was dominated by IgG-producing B cells. At the age of 8 months, a five-fold augmentation of TNP-specific B cells and of anti-TNP antibodies (AB), as compared to animals treated accordingly as adults, was observed. Clonal analysis of regulatory cells revealed 2 populations of helper (Th1 and Th2) and suppressor (Ts1 and Ts2) T cells in spleen cells (SC) of tolerized mice. In SC of mice immunized as adults, Th1 and Th2, but only one Ts populations were observed. The frequencies of Th1 and Ts1 were in the same range in animals immunized neonatally or as adults. After challenge, frequencies of regulatory cells remained constant in animals immunized as adults. But, in neonatally tolerized mice, challenge resulted in increased frequencies of Th1 and Th2; Ts1 remained constant, and concomitantly the frequency of Ts2 declined significantly. The data are interpreted as newborn tolerance being due to transient B cell anergy via receptor blockade as well as inactivation of AB-producing cells. Neither deficiency in TNP-specific help nor dominance of TNP-specific suppression is involved in maintainance of tolerance, but tolerance appears to be sustained by interference of TNP-specific regulatory cells with anti-idiotypic regulatory cells (Th2, Ts2). Supposing a system of circular network interactions, activation of anti-idiotypic clones will be counterregulated/balanced by activation of antigen-specific clones. Thus, decreasing idiotypic connectivity during life may result in overshooting reactivity of neonatally tolerized mice at older age. PMID- 2966102 TI - Suppression of the activity of natural killer-like cells by peritoneal macrophages obtained from Lewis lung tumor-bearing, Propionibacterium granulosum KP-45-treated, or normal, untreated mice. AB - Peritoneal adherent cells (PAC) obtained from Propionibacterium granulosum KP-45 treated or Lewis lung carcinoma-bearing BDF1 mice suppressed in vitro the NK-like cytotoxic activity of murine splenocytes against YAC-1 tumor target cells. Maximum inhibition occurred when suppressor and effector cells were preincubated together for 18 h, but the effect was demonstrable also when the two groups of cells were mixed only at the onset of the 4-h cytotoxic assay (i.e. without previous contact). Inhibitory cells appeared to be mostly macrophages, as judged by adherence to plastic and morphologic features, and as little as 5 to 20% of PAC, relative to the total number of co-incubated cells, were required for the clear demonstration of the effect. In addition to activated also normal, resident PAC obtained from untreated animals inhibited the NK cell-mediated cytotoxicity, but the effect was significantly pronounced only when 20% of suppressor cells were incubated overnight with effector splenocytes. The results favor the hypothesis that both functionally activated as well as resting macrophages operate as important regulators of the activity of NK cells in vivo. PMID- 2966101 TI - The bovine autologous mixed leukocyte reaction: a proliferative response of non-T cells under the control of monocytes. AB - The culture of bovine monocyte-depleted peripheral blood mononuclear cells (PBM) (responder) with irradiated autologous undepleted PBM (stimulator) resulted in a strong proliferative response, which was called the bovine autologous mixed leukocyte reaction (MLR). A comparative study was made between this autologous MLR and the bovine allogenic MLR, with respect to the inductive requirements and the nature of responding and stimulator cells. In both the autologous and allogenic MLR, the monocyte was responsible for inducing proliferation: low concentrations of purified monocytes (greater than 95% pure) induced strong proliferative responses, while monocyte-depleted PBM (less than 0.2% monocytes) were unable to induce proliferation. Both MLRs were blocked by monoclonal antibodies specific for class II antigens of the major histocompatibility complex. The magnitude of the autologous MLR was regulated by monocytes: unirradiated monocytes induced or suppressed autologous proliferative responses depending on their concentration in the culture. Proliferating cells, identified by quantifying DNA with chromomycin A3, were all within the population of large cells. In the allogenic MLR, 50% of these large cells were T cells, as defined by monoclonal antibodies. In contrast, all of the proliferating cells in the autologous MLR were non-T cells. PMID- 2966103 TI - Self versus non-self carrier in primary and secondary anti-TNP B cell response: I. Specificity of help. AB - Humoral anti-hapten responses are supposed to require carrier-specific help. Yet, "TNP-specific" help can be activated with TNP coupled to syngeneic lymphocytes. To further clarify the role of hapten-specific vs. carrier-specific helper T cells (TH) in the humoral anti-TNP response, BALB/c mice were immunized with TNP coupled to cellular or soluble self or non-self carriers, and we analyzed primary and secondary B cell responses as well as the apparent specificity of help. TNP bound to syngeneic red blood cells (sRBC), syngeneic albumin (sA) and syngeneic polyclonal or monoclonal immunoglobulin (sIg/smIg) and TNP coupled to non-self carriers initiated equivalent primary anti-TNP responses. On the other hand, a secondary anti-TNP response was only obtained with heterologous carriers or with smIg. Even with heterologous carriers or with smIg, the magnitude of the secondary anti-TNP response exceeded only slightly the amplitudes of a primary anti-TNP response. Furthermore, if animals were challenged shortly after priming, they appeared un/-hyporesponsive. In vitro analysis revealed that in the primary as well as in the secondary anti-hapten response, TNP-specific TH were involved, i.e., the primary response to sIg, sA and sRBC was exclusively due to TNP specific help, the response after priming with TNP-smIg or TNP-heterologous carrier was due to the additive effects of carrier- and TNP-specific help. Since "carrier-specific" help with smIg was independent of the antibody specificity as well as of the Ig class, we suppose that smIg activated idiotype-specific TH, which functioned like "carrier" TH. Two mechanisms were responsible for the low magnitude of the secondary anti-hapten response: competition for carrier-specific help (using heterologous carriers), and antibody (AB)-dependent suppression. PMID- 2966104 TI - Lipopolysaccharide-induced suppression of DTH-reactivity to histocompatibility antigens. I. Kinetic aspects and specificity. AB - The effects of bacterial lipopolysaccharide (LPS) on the development of DTH reactivity to alloantigens in mice were investigated. DTH to a particular set of alloantigens could be suppressed by treatment of responder mice with a single intravenous (i.v.) injection of 100 micrograms LPS and a simultaneous subcutaneous (s.c.) injection of the appropriate allogeneic spleen cells. The suppression lasted at least 60 days and affected the afferent limb of the DTH response as well as the efferent limb. The suppression could be adoptively transferred to naive syngeneic recipient mice by spleen cells, but not by immune serum, and proved to be antigen-specific. In spite of this specificity, the DTH response against unrelated "third-party" alloantigens could be suppressed as well, provided the latter were presented during the induction-phase of DTH together with the alloantigens that had been used for the induction of suppression. PMID- 2966105 TI - Non-specific suppressor cells in murine bone marrow chimeras: their possible role in GVHD-associated immunodeficiency. AB - Suppressor cells against several mitogen-induced responses were detected in the spleen of murine bone marrow chimeras, regardless of intravenous (i.v.) or intrasplenic (i.s.) bone marrow transplantation (BMT). According to the time course of the suppressor activity against Con A, PHA, and PWM, they were readily detected at 11-21 days after BMT and thereafter, either gradually decreased or remained at a plateau level. In contrast, the suppressor activity against the LPS stimulated response increased at 39-52 days as compared to 24-34 days after BMT. Characterization studies of suppressor cells early (11-21 days) after BMT revealed that those in the i.v. and i.s. chimeras were composed of host-derived plastic dish adherent and/or anti-Thy 1.2 antibody-insensitive spleen cells in general. On the contrary, those in the i.v. and i.s. chimeras that possessed severer GVHD were mainly composed of host-derived plastic dish non-adherent spleen cells. Since the suppressor activity was higher in chimeras with severe graft-versus-host disease (GVHD) than in conventional chimeras, suppressor cells against the mitogen-induced responses may be related to the immunodeficiency associated with GVHD. Particularly, plastic dish non-adherent suppressor cells may closely relate to GVHD-associated immunodeficiency as compared with plastic dish adherent suppressor cells. PMID- 2966106 TI - CR3 receptor on platelets and its role in the prostaglandin metabolic pathway. AB - The human C3R receptor, which binds C3bi, present on the surface of monocytes, granulocytes and natural killer cells, can be detected by several monoclonal antibodies, OKM1, Mo1 and Mac-1 and also by RM2.184 which detects a polymorphism of the receptor. Platelets have been considered to lack complement receptors on their cell surface; however, we now describe the detection of CR3 receptors on human platelets by radioimmunoassay using both OKM1 and RM2.184 antibodies. Using OKM1, immunoprecipitation studies with 125I-labelled platelets revealed the typical CR3 complex with an alpha chain of 165,000 daltons and beta chain of 95,000 daltons. By immunofluorescence, megakaryocytes were also found to be OKM1+. However, platelet CR3 does not merely bind C3bi, but the binding of the OKM1 antibody to platelet CR3 selectively blocks platelet functions of aggregation and serotonin release induced by arachadonic acid but not by other ligands (ristocetin, ADP, L-epinephrine, collagen and thrombin). The studies demonstrate an important role of platelet CR3 in both complement binding and in prostaglandin metabolic pathways. PMID- 2966107 TI - [Seborrheic dermatitis: use of ketoconazole cream and shampoo. Double-blind study versus placebo]. PMID- 2966109 TI - [Treatment of acne with antiandrogens: validity of treatment with spironolactone]. PMID- 2966108 TI - [Allergic dermatitis caused by contact with ethylenediamine]. PMID- 2966110 TI - [Immunohistochemical study of nodulo-cystic acne during therapy with isotretinoin. Preliminary data]. PMID- 2966111 TI - Intradermal route for prophylaxis against hepatitis B: loss of potency in relation to time. PMID- 2966112 TI - Efficacy of low-dose intradermal hepatitis B vaccine. PMID- 2966113 TI - Immunogenicity and side effects of low dose intradermally administered hepatitis B vaccine. PMID- 2966114 TI - The challenge of quality. PMID- 2966115 TI - Quality assurance through external controls. AB - Concern since the early part of this century for the quality of health care services has led to external controls in the United States that are more extensive, more intrusive, and more complex than they are elsewhere in the world. I suggest that this is the result of the decentralized nature of our health care system and the dominance of the American legal system and its adversarial orientation. This in turn has led to external control processes that focus on quantifiable, but often secondary or tangential, aspects of service that measure what is minimally acceptable rather than what is optimal. Perhaps it would be more fruitful for external control to intervene at a higher level by creating expectations for the process of quality assurance--that is, to create a system of external controls over internal controls. This, in turn, may lead to a quality assurance system focused on the more central, if more subjective, dimensions of health care. PMID- 2966116 TI - Competition and the quality of care: the importance of information. AB - Whether competitive mechanisms sacrifice quality in their efforts to control the costs of health care will depend on the ability of consumers to make informed choices of providers and the ability of providers to make informed decisions in using services in diagnosis and treatment. The availability of relevant information on the quality of providers and the effectiveness of services to both consumers and providers will determine the effects of competition on quality. We describe the types of information needed, how it can be developed, and the roles that government and the private sector must play in developing and disseminating this information. PMID- 2966117 TI - The challenges and prospects for quality assessment and assurance in ambulatory care. AB - As hospital cost containment has moved care into ambulatory settings, quality assurance requirements have followed. These requirements should focus on primary care as a means to prevent disease and limit disability while saving hospital costs. Methods for external and internal quality assurance already exist for primary care, although refinement and development of less expensive techniques is needed. Uniform quality assurance requirements that satisfy all external reviewers should be developed and applied evenhandedly to fee-for-service and prepaid providers. A top priority is the development of a uniform computerized data base for primary care encounters. PMID- 2966118 TI - Quality of care: past achievements and future challenges. AB - The heightened attention currently given to quality of care offers a unique opportunity for major advances. But to make the most of it, the greater energy and resources now available have to build on what is already known about quality of care, rather than largely ignore and therefore rediscover what has been learned in the past about, for example: how quality can be defined and measured, the relation between outcomes and the process of care, and the distinction between quality assessment and determinations of efficacy. If future efforts are rooted in what is known about these and other aspects of quality of care, they can yield substantial insights into how to improve quality, rather than simply how to measure it, and how to do it for more than inpatient hospital care. PMID- 2966119 TI - Long-term care: variations on a quality assurance theme. AB - Quality assurance in long-term care (LTC) diverges from approaches in acute care because of inherent features of LTC: the slower pace, the reliance on "low technology" services, the goals of LTC, and the multiple transitions across care sites typical of LTC users. LTC quality assurance activities should focus on outcomes, using process measures only when they are related to desirable outcomes or are intrinsic to respectful treatment of patients. Outcome measures should compare what is achieved with what can be reasonably expected. Because LTC services can be given interchangeably at various LTC sites (e.g., home care, day care, nursing homes), a data base must be developed that permits monitoring and comparing outcomes across diverse LTC programs. PMID- 2966120 TI - HMOs and the quality of care. AB - Various studies of selected health maintenance organizations (HMOs) suggest that the quality of their medical care is comparable to that delivered in the fee-for service (FFS) sector. Unfortunately, these studies are based on a small number of mature HMOs that were operating in an environment less focused on competition and cost containment than is the case today. It is the thesis of this paper that the current environment increases the importance of quality assessment and assurance for both HMO and FFS providers. I suggest that HMOs may meet the increased demand by patients and payers for quality medical care by focusing attention on population-based outcome measures. PMID- 2966121 TI - Quality-of-care research in mental health: responding to the challenge. AB - Quality-of-care research in mental health is in the developmental stages, which affords an opportunity to take an integrative approach, building on principles from efficacy, effectiveness, quality assessment, and quality assurance research. We propose an analytic strategy for designing research on the quality of mental health services using an adaptation of the structure, process, and outcome classification scheme. As a concrete illustration of our approach, we discuss research on a particular target population-patients with chronic schizophrenia. Future research should focus on developing models of treatment, establishing criteria and standards for outcomes and processes, and gathering data on community practices. PMID- 2966122 TI - Quality assessment and assurance: unity of purpose, diversity of means. AB - In this paper I attempt to advance our understanding and appreciation of the several contributions to this anthology by placing their more important features in a unifying framework. In this way I hope to show their historical antecedents, their interrelationships, and their linkages to the central problems and purposes of quality assessment and assurance. But above all, I celebrate our rededication to quality in health care. PMID- 2966123 TI - Patient satisfaction as an indicator of quality care. AB - In this review of the theoretical and empirical work on patient satisfaction with care, the most consistent finding is that the characteristics of providers or organizations that result in more "personal" care are associated with higher levels of satisfaction. Some studies suggest that more personal care will result in better communication and more patient involvement, and hence better quality of care, but the data on these issues are weak and inconsistent. Further research is needed to measure specific aspects of medical care and the ways in which patient reports can complement other sources of information about quality. In addition, more research on the determinants of satisfaction and the relationship between quality and satisfaction among hospitalized patients is recommended. PMID- 2966124 TI - The current interest in quality is nothing new. PMID- 2966125 TI - Outcome measurement: concepts and questions. AB - Outcome measurement-a central concept of quality of care-has both conceptual appeal and limitations as a practical assessment tool. The degree to which outcomes can be directly related to processes of care continues to be especially problematic. I view the continued debate about whether processes or outcomes are the preferable measure of quality as fundamentally unproductive, because both are needed. To strengthen our understanding of both measures in ascertaining quality of care, I suggest that work in four areas is needed: more definitive evidence of process and outcome linkages; stronger relationships between technology assessment and quality assessment; improved reliability and validity of outcome measures as screening tools; and continued development of health status measures. PMID- 2966126 TI - Is efficacy the gold standard for quality assessment? AB - "Efficacy"-medical care that is achievable under optimal conditions-is generally considered to be the appropriate standard for quality assurance. In this paper I argue that what is needed is a broadened definition of efficacy that includes the appropriateness of an intervention as well as the level of technical skill used in its provision. To establish efficacy, so defined, I suggest that well-designed nonexperimental, as well as experimental, studies be undertaken that employ registries and data banks, as well as formal clinical trials, and that decision analysis be used to synthesize the results. This will require a significant effort and investment in outcome studies of medical and surgical technologies. With growing recognition by the public that quality can be highly variable-aided by the Health Care Financing Administration's publication of hospital mortality and morbidity data-the medical care system will ignore outcome studies and quality assurance at its own peril. PMID- 2966127 TI - Assessing clinical decision making: is the ideal system feasible? AB - While caring for patients, physicians make a variety of decisions. Can current methods adequately determine whether these decisions are correct? If not, what improvements are needed? This paper begins with a review of several explicit methods to assess physician decision making. It then describes a more comprehensive system that would use Bayesian logic to assess whether a physician responded appropriately to the needs of an individual patient. Although sophisticated branching logic may be theoretically desirable, it may not be feasible. The paper concludes by proposing an explicit, potentially practical method that would judiciously use branching logic. PMID- 2966128 TI - Future policy directions for quality assurance: lessons from the Health Accounting experience. AB - More than 25 years ago I proposed a philosophic approach for the ongoing, systematic management of quality assurance in the U.S. health care system. Health Accounting, based on the premise that successful quality assurance must evolve into a professional discipline that integrates the health and the management sciences, called for an analysis of the benefits and "disbenefits" of health care services as rigorous as that used by business and industry to analyze fiscal profits and losses. In this paper, I review the origins and development of Health Accounting, assess what we have learned from our experience of the past 25 years, and discuss the implications of this experience for promoting the growth of quality assurance in the United States. PMID- 2966129 TI - Organized practice and the quality of medical care. AB - Larger, more complex medical care organizations have the inherent capability to improve the quality of the care they deliver because of the improved competency that follows higher volumes of service, the interdependence of staff, and the emergence of responsible leadership in large organizations. The potential for slackened physician-patient relationships, however, could jeopardize that quality. We suggest that professional associations can counterbalance the negative influences of large organizations. We envision that the changing political and economic environment of medical practice, along with the greater professional and public scrutiny of care in highly visible large organizations, will act together to exert pressure on organized practices to examine and demonstrate quality clinical practice. PMID- 2966130 TI - Quality and cost: choices and responsibilities. AB - Practitioners are at least obligated to provide the most effective care most efficiently. But when added benefits are small relative to added cost, practitioners may stop short of the most effective care in obedience to patient preferences. If society imposes a different standard of optimal care, I suggest that health care professionals will respond in one of three ways: oppose social intervention, adopt the social optimum, or take an intermediate position by accepting the social specification of optimal care but safeguarding the individual practitioner's role as an advocate for each patient and the profession's role as an advocate its view of the public good. PMID- 2966131 TI - Haber's syndrome and Dowling-Degos disease. PMID- 2966132 TI - Abdominal adipose tissue and serum HDL-cholesterol: association independent from obesity and serum triglyceride concentration. AB - It has been recently shown that an excess of abdominal fat was associated with reduced serum HDL-cholesterol level, supporting previous studies that have reported a relationship between fat distribution and cardiovascular risk factors. Since a negative relationship has been observed between serum triglyceride and HDL-cholesterol concentrations, the associations between body fat distribution and HDL-cholesterol level was studied with control over serum triglycerides in a sample of 429 healthy adult men. The relative distribution of subcutaneous fat, as reflected by the trunk to extremity skinfolds (T/E) ratio and the absolute amount of subcutaneous abdominal fat, obtained by the measurement of the abdominal skinfold thickness, were significantly correlated with serum triglycerides (r = 0.27 and 0.35 respectively, P less than 0.0001), with serum HDL-cholesterol (r = -0.14, P less than 0.01, and -0.26, P less than 0.0001) and with the serum HDL-cholesterol/total cholesterol ratio (r = -0.25 and -0.39, P less than 0.0001). Analysis of variance on two factors, the T/E ratio and the body mass index (BMI), revealed significant and independent effects of adiposity and relative distribution of subcutaneous fat on serum lipids and HDL-cholesterol (0.05 greater than P less than 0.001). However, when comparable analyses of variance were performed to study the respective contributions of obesity (as measured by the BMI) and the absolute amount of abdominal fat (as measured by the abdominal skinfold), most of the variance in serum triglycerides, cholesterol and HDL-cholesterol was explained by abdominal fat alone and not by the BMI. Therefore, it appeared that with the exception of the HDL-cholesterol/total cholesterol ratio, the association between obesity and serum lipids and HDL cholesterol was, in the present sample, primarily explained by the amount of abdominal fat. As reported by others, serum triglyceride level was negatively correlated with HDL-cholesterol (r = -0.33, P less than 0.0001) and the HDL cholesterol/total cholesterol ratio (r = -0.54, P less than 0.0001). After cholesterol and HDL-cholesterol scores were adjusted for the effect of triglycerides, the T/E ratio was no longer associated with serum HDL-cholesterol whereas the abdominal skinfold remained significantly correlated with serum HDL cholesterol concentration (r = -0.16, P less than 0.01). These results suggest that a portion of the association between body fat topography and serum HDL cholesterol is mediated by the effect of fat distribution on serum triglycerides.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966133 TI - Photodynamic effects of Photofrin II on cell division in human NHIK 3025 cells. AB - Human cervix carcinoma cells of the line NHIK 3025 were exposed to light after 18 h incubation with Photofrin II. After this photodynamic treatment cells in the interphase were retarded with respect to entry into mitosis for a period which increased with increasing light dose. Following the prolonged interphase, an increase in the mitotic index was observed, giving rise to a 3-fold higher level of mitotic cells compared to the control level. Staining of methanol-fixed cells with the DNA-specific dye mithramycin indicated that the increase in mitotic index was due to a prolongation of the metaphase. For all the light doses studied most of the metaphase cells could be characterized as three-group metaphases or c metaphase-like structures for the first 8 h after treatment. An approximately 10 fold increase above the control level in the number of tripolar mitoses was also observed. A 2h incubation in a Photofrin II-free medium after the 18 h incubation with Photofrin II and before light exposure reduced the fluorescence of the cells by 30 per cent. However, this wash-out period had no effect on the increase in mitotic index after light exposure. A light dose corresponding to 80 per cent survival (as assayed on asynchronous cells) was given to cells in mitosis after Photofrin II incubation. This treatment delayed more than 90 per cent of the metaphase cells from entering the anaphase for at least 1 h. Cells photodynamically treated in the anaphase and telophase entered the interphase at a similar rate as control cells. These observations indicate a temporary block in the initiation of the anaphase and a prolongation of the metaphase. A microscopic study of cells immunologically stained for beta-tubulin 1 h after photodynamic treatment indicated that the organization of the spindle apparatus was disturbed by the photodynamic treatment. Such perturbations are suggested to be the cause of the observed accumulation of cells in mitosis. PMID- 2966134 TI - Prognosis after meso-caval interposition shunt. Experience of eighty-six consecutive cases over a period of ten years. AB - From January 1st, 1975, up to January 1st, 1986, a meso-caval interposition shunt was performed in 86 patients (electively with one exception), using a 16 or 18 mm lumen Dacron prosthesis. The criteria for selection for operation were a liver volume of 1000-2500 ml, a residual portal perfusion of 15 to 30%, exclusion of an activity of the liver process and stenosis of the hepatic artery or coeliac trunk. The cause of portal hypertension was liver cirrhosis in 77 patients (89.5%), prehepatic block in five (4.5%), liver fibrosis in three patients (3.5%), and myeloproliferative disease in one patient (1%). The study represents a prospective assessment and included 52 men and 32 women with a mean age of 43 years. Intraoperative flow measurement demonstrated a residual portal perfusion in all 17 patients measured after the shunt. In-hospital mortality was 8%, late mortality 39%; the fate of four patients is unknown. The main causes of death were liver failure and hepatocellular carcinoma. The rate of encephalopathies over the whole period was 10.5%. The actual survival rate was over 70% for five and over 50% for ten years. Postoperative angiographic studies confirmed by sequential scintigraphy demonstrated a residual portal perfusion in 75% of the patients postoperatively, 60% after six months and in 38% after two years. Thus it could be demonstrated that the meso-caval interposition shunt is converted to a total shunt in the long term. The total shunt patency rate was 90%. The meso caval interposition shunt is certainly no ideal procedure but is useful in the selective or semiurgent management of patients with liver cirrhosis and portal hypertension. PMID- 2966135 TI - Modulation of in vitro immune reactions by platelet activating factor and a platelet activating factor antagonist. AB - Platelet activating factor (PAF) was found to suppress primary and secondary mixed lymphocyte reactions and BN52021, a naturally occurring PAF antagonist, blocked PAF-mediated suppression and enhanced the mixed lymphocyte reactions. The effect of delayed addition of PAF or BN52021 24 h or later after the initiation of cultures reduced the suppressive and enhancing effects, respectively. The removal of the antagonist BN52021 from mixed lymphocyte cultures up to 72 h after their initiation also was found to eliminate the potentiating effect of this antagonist. The continuous presence of PAF in mixed lymphocyte cultures used to generate cytotoxic lymphocytes suppressed the generation of effector cells while the addition of BN52021 elicited an enhanced level of cell-mediated cytotoxicity in such cultures. BN52021 enhanced the cytotoxic activity in such cultures irrespective of the presence of exogenous interleukin-2, suggesting that the antagonist-mediated enhancement is not due to the enhanced production of interleukin-2 by cells in the mixed cultures. The experiments reported here provide evidence for a role of PAF in modulating the complex interactions that take place in the initiation of cellular immune reactions. Furthermore, the results of these experiments indicate that the immunoregulatory action of PAF can be modulated by an antagonist that exerts its action independently of the production of the lymphocyte growth factor, interleukin-2. PMID- 2966136 TI - Protein tyrosine kinases, protein serine kinases, and the mechanism of action of insulin. PMID- 2966137 TI - [Symptoms of alcoholism from the viewpoint of the dermatologist]. AB - There is a wide range of dermatological symptoms that are indicative of alcoholism. In the literature little attention is paid to these manifestations. In the present study 201 alcoholics were examined before beginning treatment and were compared with 42 teetotallers. The symptoms observed and their causes are discussed. PMID- 2966138 TI - [Impetigo herpetiformis]. AB - We report the case of a 30-year-old patient who had impetigo herpetiformis during hormonal contraception and during two successive pregnancies. The diagnostic features of impetigo herpetiformis are reviewed. An association with HLA-Cw 6 is recognized. A large post-traumatic spleen cyst probably influenced the course of the disease. Postpartum therapy with oral glucocorticosteroids and etretinate was successful. PMID- 2966139 TI - Antibiotic prophylaxis in cardiac surgery: serum and tissue levels of teicoplanin, flucloxacillin and tobramycin. AB - In this study, the pharmacokinetics of teicoplanin have been studied in serum, fat and bone during and after cardiac surgery with two dose (400 and 200 mg) and three dose (400 mg each) regimens in a total of 49 patients. For comparison, 20 other patients, who had received a regimen of flucloxacillin (500 mg qds for five days) and tobramycin (1.5 mg/kg initially then 80 mg tds for three days), were similarly investigated. The lowest mean serum level of teicoplanin during operation was 6.9 mg/l in the two dose regimen and 9.7 mg/l with the three dose regimen. Mean serum levels of flucloxacillin and tobramycin fell to 7 and 1.4 mg/l respectively by the end of operation. At the end of bypass, fat washings contained a mean of 1.2 mg/l of teicoplanin and bone washings a mean of 6.3 mg/l. Mean tobramycin levels were 0.4 and 0.9 mg/l respectively and flucloxacillin less than 1 mg/l in washings from both tissues. The low levels of teicoplanin in fat may explain an excess of Gram-positive sternal infections in the accompanying clinical trial. PMID- 2966140 TI - Antibiotic prophylaxis in cardiac surgery: a prospective comparison of two dosage regimens of teicoplanin with a combination of flucloxacillin and tobramycin. AB - Teicoplanin, a new glycopeptide antibiotic, has a serum half-life of 47 h and excellent activity against Gram-positive bacteria, including methicillin resistant staphylococci, making it a potentially useful drug for cardiac surgical prophylaxis. In two prospective randomized studies, we have compared it with a broad spectrum regimen of an aminoglycoside and flucloxacillin. In the first trial, teicoplanin (400 mg on induction of anaesthesia and 200 mg 24 h later), was compared with tobramycin (80 mg tds for three days) and flucloxacillin (500 mg qds for 5 days) in 314 patients. Teicoplanin prophylaxis resulted in a significantly greater number of sternal wound infections (P less than 0.01), due to Gram-positive bacteria. Furthermore, Gram-negative bacteria were responsible for more respiratory and urinary infections after teicoplanin prophylaxis. In the second trial, comprising 203 patients, the teicoplanin dose regimen was changed to three doses of 400 mg but this did not improve the rates of infection. PMID- 2966141 TI - In-vitro activity of enoxacin against gonococcal isolates in comparison with that of five other antibiotics. AB - The in-vitro activity of enoxacin was investigated against 305 clinical isolates of Neisseria gonorrhoeae, including both penicillinase-producing strains (PPNG) and non-penicillinase producing strains (NPPNG), and compared with the activity of cefuroxime, acrosoxacin, spectinomycin, benzylpenicillin and ampicillin. Enoxacin was more active than the other antimicrobials tested, with an MIC90 value for the NPPNG organisms of 0.08 mg/l and for the PPNG organisms of 0.16 mg/l. Ten of the 305 isolates tested were resistant to spectinomycin; MICs of less than or equal to 0.08 mg/l were obtained for enoxacin against these organisms. PMID- 2966142 TI - Handicapped children and the law: children afflicted with AIDS. PMID- 2966144 TI - Structural analysis of staphylococcal bacteriophage phi 11 attachment sites. AB - The lysogenization of bacteriophage phi 11 in Staphylococcus aureus occurs by site-specific recombination. The DNA segments containing the attachment sites on the host chromosome, the phage genome, and the two junctions created by insertion of the prophage were cloned, and the nucleotide sequences were determined. The attachment sites share a very short common sequence of 10 base pairs. PMID- 2966143 TI - Modulation of Escherichia coli RecBCD activity by the bacteriophage lambda Gam and P22 Abc functions. AB - Plasmids that express the bacteriophage lambda gam gene or the P22 abc2 gene (with and without abc1) at controllable levels were placed in Escherichia coli and tested for effects on the activity of RecBCD. Like Gam, Abc2 inhibited the ATP-dependent exonuclease activity of RecBCD, apparently not by binding to DNA. However, Abc2-mediated inhibition was partial, while Gam-mediated inhibition was complete. Both Abc2 and Gam inhibited host system-mediated homologous recombination in a Chi-containing interval in the chromosome of a hybrid lambda phage; Abc2 inhibited it more strongly than Gam. Gam but not Abc2 spared a phage T4 gene 2 mutant from restriction by RecBCD; Abc2 exhibited weak sparing activity in combination with Abc1 and substantial activity in combination with both Abc1 and P22 homologous recombination function Erf. Either Gam or the combination of the lambda recombination functions Exo and Bet was sufficient to induce a mode of plasmid replication that produced linear multimers. The combination of Abc2, Abc1, and Erf also exhibited this activity. However, Erf was inactive, both by itself and in combination with Abc1; Abc2 had weak activity. These results indicate that Gam and Abc2 modulate the activity of RecBCD in significantly different ways. In comparison with lambda Gam, P22 Abc2 has a weak effect on RecBCD nuclease activity but a strong effect on its recombination-promoting activity. PMID- 2966145 TI - Purification and properties of the ATPase solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius. AB - A novel ATPase was solubilized from membranes of an acidothermophilic archaebacterium, Sulfolobus acidocaldarius, with low ionic strength buffer containing EDTA. The enzyme was purified to homogeneity by hydrophobic chromatography and gel filtration. The molecular weight of the purified enzyme was estimated to be 360,000. Polyacrylamide gel electrophoresis of the purified enzyme in the presence of sodium dodecyl sulfate revealed that it consisted of three kinds of subunits, alpha, beta, and gamma, whose molecular weights were approximately 69,000, 54,000, and 28,000, respectively, and the most probable subunit stoichiometry was alpha 3 beta 3 gamma 1. The purified ATPase hydrolyzed ATP, GTP, ITP, and CTP but not UTP, ADP, AMP, or p-nitrophenylphosphate. The enzyme was highly heat stable and showed an optimal temperature of 85 degrees C. It showed an optimal pH of around 5, very little activity at neutral pH, and another small activity peak at pH 8.5. The ATPase activity was significantly stimulated by bisulfite and bicarbonate ions, the optimal pH remaining unchanged. The Lineweaver-Burk plot was linear, and the Km for ATP and the Vmax were estimated to be 1.6 mM and 13 mumol Pi.mg.-1.min-1, respectively, at pH 5.2 at 60 degrees C in the presence of bisulfite. The chemical modification reagent, 7 chloro-4-nitrobenzo-2-oxa-1,3-diazole, caused inactivation of the ATPase activity although the enzyme was not inhibited by N,N'-dicyclohexylcarbodiimide, N-ethyl maleimide, azide or vanadate. These results suggest that the ATPase purified from membranes of S. acidocaldarius resembles other archaebacterial ATPases, although a counterpart of the gamma subunit has not been found in the latter. The relationship of the S. acidocaldarius ATPase to other ion-transporting ATPases, such as F0F1 type or E1E2 type ATPases, was discussed. PMID- 2966146 TI - Amino acid sequence of a peptide from keratan sulfate II-core protein linkage regions. AB - Keratan sulfate II was prepared from the proteolytic digest of pig nucleus pulposus proteoglycan. The polysaccharide chains containing the fragment peptides of the core protein at their reducing terminal were subjected to anhydrous HF solvolysis reaction and one of the glycopeptides from the keratan sulfate II-core protein linkage regions was isolated. The amino acid sequence of the peptide was deduced to be Ala-Pro-Ser-Pro-Gly, which is different from those reported for the attachment sites of chondroitin sulfate on core proteins from various sources. The results provided the first solid amino acid sequence for the keratan sulfate II-core protein linkage regions and suggested that the amino acid sequence of the core protein might determine the distribution of chondroitin sulfates and keratan sulfates along the core protein of the proteoglycan molecule. PMID- 2966147 TI - Effects of heparin and other acid mucopolysaccharides on the activity of a cytolytic pore-forming protein (perforin) from cytotoxic T-lymphocytes. AB - A cytolytic protein (perforin) was rapidly purified from a cell line of mouse cytotoxic T-lymphocytes (CTL) by DEAE-cellulose, heparin-Sepharose, and phenyl Sepharose chromatographies. The purified perforin was activated by heparin, the half maximal concentration being 3-10 ng/ml, depending on the calcium concentration. Other acid mucopolysaccharides, such as chondroitin sulfates A and C, keratan polysulfate, and heparin sulfate, also enhanced the lysis of erythrocytes by perforin, but the concentrations required for activation were more than 100-fold higher than that of heparin. Chondroitin, hyaluronic acid, and keratan sulfate, however, had no effect on the perforin activity. It was suggested that heparin potentiates the lytic activity of perforin and acid mucopolysaccharides may actually be involved in target cell lysis by CTL. PMID- 2966148 TI - Separation of SH-modified myosin subfragment-1 (A1) isozyme into two distinct equimolar fractions by an affinity chromatography. AB - Our previous kinetic studies indicated that SH-modified myosin subfragment-1 A1 isozyme (S1(A1] contains at least two different types of active sites (Emoto, Y., Kawamura, T., & Tawada, K. (1985) J. Biochem. 98, 735-745). In those studies we have modified highly reactive SH-groups in S1(A1) with thimerosal. In this work, we separated the modified S1(A1) into two equimolar fractions by affinity chromatography with agarose-ADP. For the separation, Mg2+ in the elution buffer was indispensable. Although the two fractions appeared to have the same number of modified SH-groups per mol of S1, they had different enzymic and fluorescent properties. SH-modification with an excess of thimerosal for a much longer duration did not change any of the results: not the chromatographic profile, the properties of the two fractions, nor the number of modified SH-groups. Hence the two different populations were not generated by incomplete modification. After reduction with dithiothreitol, however, the differences between the two fractions disappeared. When we separately re-modified the reduced fractions and re chromatographed them, in each case we again obtained two fractions, which had the same properties as the two fractions obtained from the original modification with thimerosal. These results demonstrate that the active site heterogeneity in SH modified S1(A1) had no intrinsic origin in the unmodified S1: it was introduced by the SH-modification, but by an unknown mechanism(s) other than incomplete modification. PMID- 2966149 TI - Steady-state kinetic study on the inhibition of the adenosinetriphosphatase activity of dynein from Tetrahymena cilia by glycerol. AB - The characteristics of glycerol-induced inhibition of the dynein ATPase extracted from Tetrahymena cilia were investigated. Fifty percent inhibition was observed at about 15% (v/v) glycerol with the 22S dynein Mg-ATPase. Ethylene glycol was equally inhibitory, while sucrose, a kind of polyol, was less effective. The glycerol-induced inhibition of the 22S dynein Mg-ATPase was not influenced by pH or by raising the ionic strength of the assay solution. An aqueous glycerol solution treated with anion or cation exchanger or charcoal was equally inhibitory to a non-treated solution. The inhibition was most likely to be due to glycerol or ethylene glycol itself, not to a contaminant. The inhibition of the 22S dynein Mg-ATPase was apparently noncompetitive: only the Vmax was reduced without a significant change in the apparent Km. The dynein ATPase is known to be inhibited potently by vanadate. Glycerol reduced the sensitivity of the dynein ATPase to the vanadate-induced inhibition. Glycerol exhibited a decelerating effect on the rate of the oxygen exchange between phosphate and water catalyzed by 22S dynein in the presence of ADP and Mg2+. If it is assumed that the rate constants of the ATP hydrolysis step are not affected by glycerol, it may be implied that the phosphate release from the E.ADP.P1 intermediate was decelerated by glycerol and that the deceleration of the phosphate release paralleled the reduction of the overall ATPase activity over a wide range of glycerol concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966150 TI - Biochemical evidence supporting a mechanism for cap-independent and internal initiation of eukaryotic mRNA. AB - The role of eukaryotic initiation factor (eIF)4B in translation is somewhat uncertain, although it appears to stimulate a variety of activities of eIF-4A and eIF-4F. Using the model RNA-dependent ATP hydrolysis assay, the ability of eIF-4B to stimulate eIF-4A and eIF-4F was investigated. The most dramatic effect of eIF 4B is to increase the affinity of eIF-4A for RNA; no effect is seen on the affinity of eIF-4A for ATP. This is not the case for eIF-4F where stimulation occurs primarily through an increase in Vmax and not a change in the affinity for RNA. The finding that eIF-4A and eIF-4B can bind to an mRNA (lacking in secondary structure), with essentially the same degree of effectiveness and affinity as would occur for natural mRNAs in the presence of eIF-4A, eIF-4B, and eIF-4F, suggests a possible role for eIF-4A and eIF-4B in both cap-independent and internal initiation. PMID- 2966151 TI - K-252a, a novel microbial product, inhibits smooth muscle myosin light chain kinase. AB - Effects of K-252a, (8R*, 9S*, 11S*)-(-)-9-hydroxy-9-methoxycarbonyl-8-methyl 2,3,9,10-tetrahydro-8, 11-epoxy-1H,8H,11H-2,7b,11a-triazadibenzo[a,g]cycloocta [cde]trinden-1-one, purified from the culture broth of Nocardiopsis sp., on the activity of myosin light chain kinase were investigated. 1) K-252a (1 x 10(-5) M) affected three characteristic properties of chicken gizzard myosin-B, natural actomyosin, to a similar degree: the Ca2+-dependent activity of ATPase, superprecipitation, and the phosphorylation of the myosin light chain. 2) K-252a inhibited the activities of the purified myosin light chain kinase and a Ca2+ independent form of the enzyme which was constructed by cross-linking of myosin light chain kinase and calmodulin using glutaraldehyde. The degrees of inhibition by 3 x 10(-6) M K-252a were 69 and 48% of the control activities with the purified enzyme and the cross-linked complex, respectively. Chlorpromazine (3 x 10(-4) M), a calmodulin antagonist, inhibited the native enzyme, but not the cross-linked one. These results suggested that K-252a inhibited myosin light chain kinase by direct interaction with the enzyme, whereas chlorpromazine suppressed the enzyme activation by interacting with calmodulin. 3) The inhibition by K-252a of the cross-linked kinase was affected by the concentration of ATP, a phosphate donor. The concentration causing 50% inhibition was two orders magnitude lower in the presence of 100 microM ATP than in the presence of 2 mM ATP. 4) Kinetic analyses using [gama-32P]ATP indicated that the inhibitory mode of K-252a was competitive with respect to ATP (Ki = 20 nM). These results suggest that K-252a interacts at the ATP-binding domain of myosin light chain kinase. The direct action of the compound on the enzyme would explain the multivarious inhibition of myosin ATPase, of superprecipitation, and of the contractile response of smooth muscle. PMID- 2966152 TI - Structural analogs of human insulin-like growth factor I with reduced affinity for serum binding proteins and the type 2 insulin-like growth factor receptor. AB - Four structural analogs of human insulin-like growth factor I (hIGF-I) have been prepared by site-directed mutagenesis of a synthetic IGF-I gene and subsequent expression and purification of the mutant protein from the conditioned media of transformed yeast. [Phe-1,Val1,Asn2, Gln3,His4,Ser8, His9,Glu12,Tyr15,Leu16]IGF-I (B-chain mutant), in which the first 16 amino acids of hIGF-I were replaced with the first 17 amino acids of the B-chain of insulin, has greater than 1,000-, 100 , and 2-fold reduced potency for human serum binding proteins, the rat liver type 2 IGF receptor, and the human placental type 1 IGF receptor, respectively. The B chain mutant also has 4-fold increased affinity for the human placental insulin receptor. [Gln3,Ala4]IGF-I has 4-fold reduced affinity for human serum binding proteins, but is equipotent to hIGF-I at the types 1 and 2 IGF and insulin receptors. [Tyr15,Leu16]IGF-I has 4-fold reduced affinity for human serum binding proteins and 10-fold increased affinity for the insulin receptor. This peptide is also equipotent to hIGF-I at the types 1 and 2 IGF receptors. The peptide in which these four-point mutations are combined, [Gln3,Ala4,Tyr15,Leu16]IGF-I, has 600-fold reduced affinity for the serum binding proteins. This peptide has 10 fold increased potency for the insulin receptor, but is equipotent to hIGF-I at the types 1 and 2 IGF receptors. All four of these mutants stimulate DNA synthesis in the rat vascular smooth muscle cell line A10 with potencies reflecting their potency at the type 1 IGF receptor. These studies identify some of the domains of hIGF-I which are responsible for maintaining high affinity binding with the serum binding protein and the type 2 IGF receptor. In addition, these peptides will be useful in defining the role of the type 2 IGF receptor and serum binding proteins in the physiological actions of hIGF-I. PMID- 2966153 TI - Dicyclohexylcarbodiimide interaction with sarcoplasmic reticulum. Inhibition of Ca2+ efflux. AB - Dicyclohexylcarbodiimide (DCCD), a hydrophobic carboxyl reagent, inhibited Ca2+ release from Ca2+-loaded sarcoplasmic reticulum vesicles, induced by elevated pH, tetraphenylboron, ATP + Pi, or membrane modification with acetic anhydride. Under the conditions used, the same concentrations of DCCD were required for inhibition of Ca2+ release, Ca2+-ATPase activity, and Ca2+ uptake. On the other hand, free Ca2+ or alkaline pH prevented the inhibition by DCCD of Ca2+-ATPase and coupled Ca2+ transport but not that of Ca2+ release. Moreover, several hydrophilic carboxyl reagents inhibited Ca2+-ATPase but not Ca2+ release. We suggest that a carboxyl residue(s), located in a hydrophobic region of a protein(s), is involved in the control of Ca2+ release, where DCCD interaction with this group blocks Ca2+ release. This group is distinct from the one involved in the inhibition of Ca2+-ATPase. DCCD also inhibited [3H]ryanodine binding to junctional sarcoplasmic reticulum membranes. The presence of Ca2+ or an alkaline pH only slightly affects the degree of inhibition of ryanodine binding by DCCD. Incubation of the membranes with [14C]DCCD resulted in labeling of 350-, 170-, 140-, 53-, and 30 kDa proteins in addition to the Ca2+-ATPase. The involvement of one or all of the DCCD-labeled proteins in Ca2+ release and ryanodine binding is discussed. PMID- 2966154 TI - Interactions of low density lipoprotein2 and other apolipoprotein B-containing lipoproteins with lipoprotein(a). AB - Studies were undertaken to investigate potential interactions among plasma lipoproteins. Techniques used were low density lipoprotein2 (LDL2)-ligand blotting of plasma lipoproteins separated by nondenaturing 2.5-15% gradient gel electrophoresis, ligand binding of plasma lipoproteins by affinity chromatography with either LDL2 or lipoprotein(a) (Lp(a)) as ligands, and agarose lipoprotein electrophoresis. Ligand blotting showed that LDL2 can bind to Lp(a). When apolipoprotein(a) was removed from Lp(a) by reduction and ultracentrifugation, no interaction between LDL2 and reduced Lp(a) was detected by ligand blotting. Ligand binding showed that LDL2-Sepharose 4B columns bound plasma lipoproteins containing apolipoproteins(a), B, and other apolipoproteins. The Lp(a)-Sepharose column bound lipoproteins containing apolipoprotein B and other apolipoproteins. Furthermore, the Lp(a) ligand column bound more lipoprotein lipid than the LDL2 ligand column, with the Lp(a) ligand column having a greater affinity for triglyceride-rich lipoproteins. Lipoprotein electrophoresis of a mixture of LDL2 and Lp(a) demonstrated a single band with a mobility intermediate between that of LDL2 and Lp(a). Chemical modification of the lysine residues of apolipoprotein B (apoB) by either acetylation or acetoacetylation prevented or diminished the interaction of LDL2 with Lp(a), as shown by both agarose electrophoresis and ligand blotting using modified LDL2. Moreover, removal of the acetoacetyl group from the lysine residues of apoB by hydroxylamine reestablished the interaction of LDL2 with Lp(a). On the other hand, blocking of--SH groups of apoB by iodoacetamide failed to show any effect on the interaction between LDL2 and Lp(a). Based on these observations, it was concluded that Lp(a) interacts with LDL2 and other apoB-containing lipoproteins which are enriched in triglyceride; this interaction is due to the presence of apolipoprotein(a) and involves lysine residues of apoB interacting with the plasminogen-like domains (kringle 4) of apolipoprotein(a). Such results suggest that Lp(a) may be involved in triglyceride-rich lipoprotein metabolism, could form transient associations with apoB-containing lipoproteins in the vascular compartment, and alter the intake by the high affinity apoB, E receptor pathway. PMID- 2966155 TI - Identification of the phosphorylation sites in early region 1A proteins of adenovirus type 5 by amino acid sequencing of peptide fragments. AB - We have mapped the positions of three of the phosphorylation sites on the 289 and 243 residue (289R and 243R) early region 1A (E1A) proteins of human adenovirus type 5 (Ad5). These proteins, which play roles in both transcriptional control and oncogenic transformation, have identical sequences except for the presence in 289R of 46 additional internal amino acids. Phosphorylation was detected exclusively at serine residues. E1A proteins purified from [35S]methionine- or [32P]orthophosphate-labeled Ad5-infected cells were digested with trypsin, and two phosphopeptides were isolated by reverse-phase chromatography and subjected to automated Edman degradation. The major species was shown to contain a single phosphorylation site at Ser-219. The second phosphopeptide was shown to contain at least one phosphorylation site at Ser-231. A third phosphorylated tryptic peptide could not be eluted from the column but was isolated using an E1A specific rat monoclonal antibody. Following subcleavage by Staphylococcus aureus V-8 protease, this peptide was shown to contain at least one phosphorylation site at Ser-89. The present data indicate that both the 289R and 243R E1A proteins are phosphorylated at the same sites, at least one in the amino terminal half of the molecule, and at least two toward the carboxyl terminus. PMID- 2966157 TI - Neurospora tryptophan synthase. Characterization of the pyridoxal phosphate binding site. AB - Tryptophan synthase, which catalyzes the final step of tryptophan biosynthesis, is a multifunctional protein that requires pyridoxal phosphate for two of its three distinct enzyme activities. Tryptophan synthase from Neurospora crassa, a homodimer of two 75-kDa subunits, was shown to bind 1 mol of pyridoxal phosphate/mol of subunit with a calculated dissociation constant for pyridoxal phosphate of 1.1 microM. The spectral properties of the holoenzyme, apoenzyme, and reconstituted holoenzyme were characterized and compared to those previously established for the heterotetrameric (alpha 2 beta 2) enzyme from Escherichia coli. The Schiff base formed between pyridoxal phosphate and the enzyme was readily reduced by sodium borohydride, but not sodium cyanoborohydride. The active site residue that binds pyridoxal phosphate, labeled by reduction of the Schiff base with tritium-labeled sodium borohydride, was determined to be lysine by high performance liquid chromatography analysis of the protein hydrolysate. A 5400-dalton peptide containing the reduced pyridoxal phosphate moiety was generated by cyanogen bromide treatment, purified and sequenced. The sequence is 85% homologous with the corresponding sequence obtained for yeast tryptophan synthase (Zalkin, H., and Yanofsky, C. (1982) J. Biol. Chem. 257, 1491-1500); the lysine derivatized by pyridoxal phosphate is located at the same relative position as that in the yeast and E. coli enzymes. PMID- 2966156 TI - Effects of phosphorylation of light chain residues threonine 18 and serine 19 on the properties and conformation of smooth muscle myosin. AB - Smooth muscle myosin can be phosphorylated by myosin light chain kinase at the serine 19 and threonine 18 residues of the two 20,000-dalton light chains (Ikebe, M., Hartshorne, D. J., and Elizinga, M. (1986) J. Biol. Chem. 261, 36-39). These studies with myosin and heavy meromyosin (HMM) compare the effects induced by phosphorylation of serine 19 (M2P and HMM2P) and serine 19 plus threonine 18 (M4P and HMM4P). Formation of M4P altered the KCl dependence of viscosity and Mg2+ ATPase and higher values were maintained at lower ionic strengths, compared to M2P or dephosphorylated myosin (Mo). This is consistent with the stabilization of the 6 S conformation. The tendency for aggregation, as judged by light scattering, followed the sequence M4P greater than M2P greater than Mo. Filaments formed with M4P were more resistant to dissociation by ATP compared to filaments of M2P. Phosphorylation of HMM2P doubled Vmax of actin-activated ATPase with little effect on the apparent affinity for actin. The Mg2+-ATPase of HMM4P exhibited a higher activity at low ionic strength compared to HMM2P and HMMo. Hydrodynamic differences were detected at low ionic strength in the presence of ATP by sedimentation velocity measurements with HMM4P, HMM2P, and HMMo. Proteolysis by papain indicated an increased susceptibility of the head-neck junction of HMM4P compared to HMM2P. These data suggest that the phosphorylation of threonine 18 in addition to serine 19 change the conformation of myosin and HMM and this is associated with altered biological properties. PMID- 2966158 TI - Mafenide-induced pseudochondritis. AB - Wound infections following burns of the ear can result in the devastating complication of chondritis, requiring resection of cartilage. To prevent this, it has become common practice to dress the burned ear with mafenide acetate. We have observed six hypersensitivity reactions to the mafenide that occurred following several weeks of continuous use of the drug. The reaction mimics chondritis, causing edematous, erythematous, pruritic ears with a profuse serous exudate. There is no associated fever, systemic signs, or pain on motion of the cartilage. Treatment consists of stopping the mafenide. Recovery occurs within 72 hours. Differentiating between chondritis, with its required surgical and antibiotic treatment, and a hypersensitivity reaction is necessary to avoid further iatrogenic injury. PMID- 2966159 TI - Fungal microabscesses in immuno-suppressed patients--CT appearances. AB - Disseminated fungal infections in leukemic patients treated with cytotoxic drugs are an increasing problem. Confirmation of such infection either by clinical or laboratory examination can be extremely difficult. The findings in four such patients, using computed tomography, are described here. The presence of small non-enhancing lesions in the liver, spleen, or kidney in such patients is strongly suggestive of deep-seated fungal infections. PMID- 2966160 TI - Radiologic features of Deve's accessory lobe. AB - In 10-15% of the population a fissure, situated at the same level as the minor fissure, partially or completely separates the superior segment from the basal segments of the lower lobe, more commonly on the right. A review of 25 patients showed that the accessory lobe produced by this fissure has a characteristic appearance when consolidated or collapsed. On the frontal view the consolidation is sharply marginated inferiorly by the accessory fissure, and it does not obscure the mediastinal margin. On the lateral view it has a triangular shape bounded by the oblique and accessory fissures. When there is loss of volume the accessory fissure is elevated. PMID- 2966162 TI - The radiology of sacral cysts. AB - The radiological findings in 10 patients with sacral cysts were retrospectively reviewed and classified. The cysts were an incidental finding on computed tomography (CT) in four patients. The expansion of sacral foramina or the sacral canal as seen on plain films suggested the diagnosis in three. In only two of the five patients who had myelograms did the cysts fill with contrast. In eight, CT showed remodelling and expansion of the sacral foramina, or the canal, or both, by a homogeneous mass with a density of 5-20 Hounsfield units. One of the patients underwent magnetic resonance imaging which confirmed that the lesion was fluid-filled. We found that sacral cysts can be either symptomatic or asymptomatic, that they may or may not communicate with the subarachnoid space, and that they have a characteristic CT appearance. PMID- 2966161 TI - Clinical-radiologic-pathologic conference: a family with lung nodules discovered by radiographic survey. PMID- 2966163 TI - Ultrasonography and CT of multiple bile duct hamartomas simulating dilated intrahepatic ducts. PMID- 2966164 TI - Duplication of the colon: varied presentations of a rare congenital anomaly. AB - Duplication of the colon is a rare congenital anomaly which may produce diagnostic difficulties for both clinician and radiologist. We report three patients who illustrate the spectrum of clinical and radiographic features which may be encountered. Radiology is essential in assessing the full extent of this condition including its various presentations, associated congenital anomalies, and anticipated complications. PMID- 2966165 TI - Magnetic resonance imaging, computed tomography, and radionuclide scintigraphy in detection of liver metastases. AB - A series of 100 patients with suspected hepatic metastases was studied with magnetic resonance (MR), unenhanced computed tomography (CT), and radionuclide (RN) scintigraphy. Each set of images was read by three clinicians using a five point scale to allow receiver operating characteristic (ROC) analysis using truth data derived from clinical review. Performance was measured by the areas under the ROC curves (0.940 +/- .018 for MR, 0.951 O +/- .013 for CT and 0.943 +/- .013 for RN) which were statistically not significantly different. We conclude that at their present level of development these three diagnostic examinations have equivalent performance and that MR is not superior in the detection of hepatic metastases. PMID- 2966166 TI - Information explosion in radiology. AB - We studied the increase in the number of articles in the radiologic literature in the years 1970, 1978, and 1985 using the Medlars program which provides reproducibility, a recognized bibliographic pool, and a wide coverage of the medical literature. A dramatic increase in the number of articles was found; the increase has both positive and negative implications for the profession. A similar analysis of monographs using the Catline file and the same subject headings also showed a significant increase in the volume of material relating to certain radiologic subspecialties. The problems arising from the attempt to identify and extract relevant information from this overwhelming mass of radiological data should be addressed. PMID- 2966167 TI - Chernobyl--the radiological impact on Canada. AB - On 26 April 1986, an accident at a Ukrainian nuclear reactor at Chernobyl triggered the release of large quantities of fission products into the atmosphere. After 7 May 1986 measurable quantities of ruthenium-103, iodine-131, cesium-134, and cesium-137 were detected in environmental sampling carried out in all regions of Canada. Maximum airborne concentrations for each radionuclide were of the order of a few mBq.m-3 and contaminated milk samples on average contained less than 1Bq.L-1 of iodine-131 and cesium-137. The mean value of the effective dose equivalent for an adult Canadian in the two months following the accident is calculated to be 0.28 microSv. As this total radiation dose is about 10(-33) of the dose from natural background during the same period, the resultant radiological detriment is concluded to be negligible. PMID- 2966168 TI - Construction of three-dimensional compensators using a simplified moire camera. AB - We describe a simple technique for fabrication of tissue compensators using a simplified moire contouring camera mounted on a commercial simulator. It is shown that high-quality optical enlargement of moire photographs can be employed to make tissue compensators quickly using 0.4 mm lead sheets. Phantom measurement shows that this technique accurately compensates an irregularly shaped surface. PMID- 2966169 TI - Opinion: contrast media, ionic and non-ionic, a personal view. PMID- 2966170 TI - Eosinophilic prostatitis simulating invasive prostatic carcinoma: CT appearance. AB - We report the first description of the computed tomographic findings in a patient with eosinophilic prostatitis, a rare inflammatory disorder of the prostate. The appearances were indistinguishable from advanced invasive prostatic cancer but showed almost complete resolution following corticosteroid therapy. PMID- 2966171 TI - Contrast layering in a choledochal cyst: a new CT observation. AB - A choledochal cyst is an uncommon congenital abnormality and may present with a wide range of clinical symptoms. I here report a new finding from the use of computed tomography, namely layering of intravenous contrast agent within the cyst. This direct sign gives a definitive diagnosis when clinical or other imaging features may not clearly establish the diagnosis. PMID- 2966172 TI - Permanent subcutaneous acupuncture needles: radiographic manifestations. AB - Metallic acupuncture needles are occasionally placed permanently in subcutaneous tissues. We report the radiographic appearances of these needles in two patients, each showing many dorsal subcutaneous needles in parallel longitudinal lines along traditionally delineated meridians between the neck and buttocks. PMID- 2966173 TI - The diagnosis of a pleural lipoma by CT and fine-needle biopsy to avoid thoracotomy. AB - We report a patient with a rare benign pleural lipoma diagnosed using computed tomography and fine-needle biopsy, thus avoiding unnecessary exploratory surgery. PMID- 2966175 TI - The rapid evolution of a large mycotic aneurysm of the abdominal aorta. AB - Computed tomography (CT) and ultrasonography (US) have been shown to be effective, noninvasive methods for the detection of established mycotic aortic aneurysms. We report a patient in whom CT and US were performed immediately before and after the development of a large mycotic abdominal aortic aneurysm, providing a unique opportunity to observe its rapid evolution. PMID- 2966174 TI - Radiologic treatment of a patient with the "steakhouse syndrome": case report. AB - A method of treating the "steakhouse syndrome" (esophageal obstruction due to meat impaction), employing air insufflation via a nasoesophageal tube with esophageal hypotonia, is described as applied in one patient. The technique is a modification of tube esophagography and can be attempted when the combination of an effervescent agent and a hypotonic drug has been unsuccessful. PMID- 2966176 TI - Fracture of the polyethylene acetabular cup in total hip arthroplasties. AB - Fracture of the acetabular cup is an uncommon complication in total hip arthroplasties, with only six examples described in the orthopedic literature. A patient is presented to illustrate the plain radiographic findings of acetabular cup fracture and to discuss the possible mechanisms involved. PMID- 2966177 TI - Benign, atraumatic, bronchoesophageal fistulae. AB - We report two patients with bronchoesophageal fistulae resulting from benign causes. The first resulted from erosion of an acquired middle esophageal diverticulum. The second had the prolonged clinical course of a presumed congenital fistula. The importance of barium examinations in evaluating chronic respiratory complaints is emphasized. PMID- 2966178 TI - Pyogenic spondylodiscitis with disc space widening. PMID- 2966180 TI - A monoclonal antibody against a laminin-heparan sulfate proteoglycan complex perturbs cranial neural crest migration in vivo. AB - INO (inhibitor of neurite outgrowth) is a monoclonal antibody that blocks axon outgrowth, presumably by functionally blocking a laminin-heparan sulfate proteoglycan complex (Chiu, A. Y., W. D. Matthew, and P. H. Patterson. 1986. J. Cell Biol. 103: 1382-1398). Here the effect of this antibody on avian neural crest cells was examined by microinjecting INO onto the pathways of cranial neural crest migration. After injection lateral to the mesencephalic neural tube, the antibody had a primarily unilateral distribution. INO binding was observed in the basal laminae surrounding the neural tube, ectoderm, and endoderm, as well as within the cranial mesenchyme on the injected side of the embryo. This staining pattern was indistinguishable from those observed with antibodies against laminin or heparan sulfate proteoglycan. The injected antibody remained detectable for 18 h after injection, with the intensity of immuno-reactivity decreasing with time. Embryos ranging from the neural fold stage to the 9-somite stage were injected with INO and subsequently allowed to survive for up to 1 d after injection. These embryos demonstrated severe abnormalities in cranial neural crest migration. The predominant defects were ectopic neural crest cells external to the neural tube, neural crest cells within the lumen of the neural tube, and neural tube deformities. In contrast, embryos injected with antibodies against laminin or heparan sulfate proteoglycan were unaffected. When embryos with ten or more somites were injected with INO, no effects were noted, suggesting that embryos are sensitive for only a limited time during their development. Immunoprecipitation of the INO antigen from 2-d chicken embryos revealed a 200-kD band characteristic of laminin and two broad smears between 180 and 85 kD, which were resolved into several bands at lower molecular mass after heparinase digestion. These results indicate that INO precipitates both laminin and proteoglycans bearing heparan sulfate residues. Thus, microinjection of INO causes functional blockage of a laminin-heparan sulfate proteoglycan complex, resulting in abnormal cranial neural crest migration. This is the first evidence that a laminin-heparan sulfate proteoglycan complex is involved in aspects of neural crest migration in vivo. PMID- 2966179 TI - Characterization of the thermotolerant cell. II. Effects on the intracellular distribution of heat-shock protein 70, intermediate filaments, and small nuclear ribonucleoprotein complexes. AB - Here we further characterize a number of properties inherent to the thermotolerant cell. In the preceding paper, we showed that the acquisition of the thermotolerant state (by a prior induction of the heat-shock proteins) renders cells translationally tolerant to a subsequent severe heat-shock treatment and thereby results in faster kinetics of both the synthesis and subsequent repression of the stress proteins. Because of the apparent integral role of the 70-kD stress proteins in the acquisition of tolerance, we compared the intracellular distribution of these proteins in both tolerant and nontolerant cells before and after a severe 45 degrees C/30-min shock. In both HeLa and rat embryo fibroblasts, the synthesis and migration of the major stress-induced 72-kD protein into the nucleolus and its subsequent exit was markedly faster in the tolerant cells as compared with the nontolerant cells. Migration of preexisting 72-kD into the nucleolus was shown to be dependent upon heat-shock treatment and independent of active heat-shock protein synthesis. Using both microinjection and immunological techniques, we observed that the constitutive and abundant 73-kD stress protein similarly showed a redistribution from the cytoplasm and nucleus into the nucleolus as a function of heat-shock treatment. We show also that other lesions that occur in cells after heat shock can be prevented or at least minimized if the cells are first made tolerant. Specifically, the heat-induced collapse of the intermediate filament cytoskeleton did not occur in cells rendered thermotolerant. Similarly, the disruption of intranuclear staining patterns of the small nuclear ribonucleoprotein complexes after heat-shock treatment was less apparent in tolerant cells exposed to a subsequent heat-shock treatment. PMID- 2966182 TI - Atrial natriuretic factor is detectable in human cerebrospinal fluid. AB - To determine whether atrial natriuretic factor (ANF) is detectable in human cerebrospinal fluid (CSF), I measured ANF in CSF samples obtained from subjects undergoing myelography for various neurological disorders. Immunoreactive ANF, measured by RIA, ranged from undetectable (less than 9.7) to 36 pmol/L. All measurable samples diluted in parallel in the RIA. Reverse phase high pressure liquid chromatography and subsequent RIA confirmed the presence of ANF, with most of the immunoreactivity eluting at the position of standard human ANF-(99-126). Simultaneous plasma ANF levels were measured in most individuals and the plasma concentrations were less than those in CSF; most of the immunoreactivity in plasma also eluted at the position of human ANF-(99-126). Thus, ANF is present in CSF in substantial amounts in some subjects with neurological disorders. Further, the CSF levels exceeded the circulating plasma ANF levels in most subjects. These results suggest that ANF in CSF is at least partially derived from production in the brain and/or is concentrated in the CSF. PMID- 2966181 TI - Glycoprotein Ic-IIa functions as an activation-independent fibronectin receptor on human platelets. AB - Soluble fibronectin binds specifically to glycoprotein (GP) IIb-IIIa on thrombin activated platelets, and this binding is not observed with platelets of patients with Glanzmann's thrombasthenia (GT) which lack GPIIb-IIIa. Here we report that GT platelets retain the ability to interact with fibronectin-coated surfaces. Adhesion to fibronectin does not require platelet activation and is inhibited by soluble fibronectin, antibodies specific for fibronectin, peptides containing the sequence Arg-Gly-Asp and polyclonal antibodies specific for band 3 of the chicken embryo fibroblast fibronectin receptor (anti-band 3). Using anti-band 3, we have purified a second fibronectin receptor from human platelets, a heterodimer composed of glycoproteins previously designated GPIc and GPIIa. The GPIc-IIa complex is found on both GT and normal platelets and appears to be identical to the GP138 kD-GP160 kD complex recently immunopurified by Giancotti et al. (1986. Exp. Cell Res. 163:47-62) and by Sonnenberg et al. (1987. J. Biol. Chem. 268:10376-10383). In this report, we provide the first evidence that GPIc-IIa actually mediates adhesion of platelets to fibronectin-coated surfaces. GPIc-IIa thus represents a second functional fibronectin receptor, distinct from GPIIb IIIa, that is largely responsible for the adhesion of nonactivated platelets to fibronectin-coated surfaces. PMID- 2966183 TI - Hormonal and clinical effects of ketoconazole in hirsute women. AB - The effects of ketoconazole on pituitary-ovarian function and adrenal function were evaluated in nine hirsute women treated with 400-1200 mg/day for 1-6 months. High dose (800-1200 mg/day) ketoconazole treatment decreased serum androstenedione, dehydroepiandrosterone, and testosterone (T) concentrations, while that of 17 alpha-hydroxyprogesterone increased, suggesting a steroidogenic block at the level of 17,20-desmolase. The decreased serum T and increased sex hormone-binding globulin concentrations led to a significant decrease in the free androgen index. Serum estradiol, cortisol, and dehydroepiandrosterone sulfate concentrations did not change. The serum LH concentration and the LH to FSH ratio increased during treatment, suggesting a negative feedback effect of T on pituitary LH secretion. The hormonal changes that occurred during high dose ketoconazole therapy persisted during subsequent low dose (400 mg/day) treatment. The therapeutic effect of ketoconazole on hirsutism manifested itself at 6 months. We conclude that ketoconazole reduces excessive androgen production in a dose-dependent manner and decreases the free androgen index in hirsute women; these changes are accompanied by significant alleviation of hirsutism. PMID- 2966185 TI - Excretion of loratadine in human breast milk. AB - The excretion of loratadine, a new nonsedating antihistamine, into human breast milk was studied in six lactating nonpregnant volunteers. Each volunteer received one 40-mg loratadine capsule. Milk and blood were collected before and at specified times (to 48 hours) after dosing. Plasma and milk loratadine concentrations were determined by a specific radioimmunoassay, and those of an active but minor metabolite, descarboethoxyloratadine, by high performance liquid chromatography (HPLC). Breast milk concentration-time curves of both loratadine and descarboethoxyloratadine paralleled the plasma concentration-time curves. For loratadine, the plasma Cmax was 30.5 ng/mL at 1.0 hour after dosing and the milk Cmax was 29.2 ng/mL in the 0 to 2 hour collection interval. Through 48 hours, the loratadine milk-plasma AUC ratio was 1.2 and 4.2 micrograms of loratadine was excreted in breast milk, which was 0.010% of the administered dose. For descarboethoxyloratadine, the plasma Cmax was 18.6 ng/mL at 2.2 hours after dosing, whereas the milk Cmax was 16.0 ng/mL, which was in the 4 to 8-hour collection interval. Through 48 hours, the mean milk-plasma descarboethoxyloratadine AUC ratio was 0.8 and a mean of 6.0 micrograms of descarboethoxyloratadine (7.5 micrograms loratadine equivalents) were excreted in the breast milk, or 0.019% of the administered loratadine dose. Thus, a total of 11.7 micrograms loratadine equivalents or 0.029% of the administered dose were excreted as loratadine and its active metabolite. A 4-kg infant ingesting the loratadine and descarboethoxyloratadine excreted would receive a dose equivalent to 0.46% of the loratadine dose received by the mother on a mg/kg basis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966184 TI - Use of the MMPI and MCMI in predicting outcome of lumbar laminectomy. AB - This study compared the abilities of the MMPI and Millon Clinical Multiaxial Inventory (MCMI), using discriminant analyses, to predict outcome after lumbar laminectomy for chronic back pain. Sixty-nine males and 60 females with lumbar discogenic disease completed MMPIs and MCMIs before surgery and were classified as either having good or fair/poor surgical outcomes based on self-reported pain relief, return to work, restriction of activities, and medication use. Results showed both the MMPI and MCMI to have moderate ability to predict surgery outcome. A slightly higher classification hit rate was obtained with both instruments when age, sex, employment status, and presence of compensation/litigation issues also were entered into the prediction equation. Results indicate the need for caution in using either instrument to make predictions of surgery outcome in individual cases. PMID- 2966186 TI - Dentists' liability for handicap discrimination against AIDS patients. PMID- 2966187 TI - Evaluation and management of the dental patient with cancer I: Complications associated with chemotherapy or bone marrow transplantation. PMID- 2966188 TI - Coping with AIDS: issues of accountability in the dental office. PMID- 2966189 TI - Caregiver interactions with autistic children. AB - Caregiver interactions with young autistic children were contrasted with those involving caregivers and developmentally matched mentally retarded and normal infants. Caregivers of autistic children were similar to other caregivers in their responsiveness to child nonverbal communication bids and in their engagement in mutually sustained play. Caregivers of autistic children were similar to caregivers of mentally retarded children in their greater use of control strategies. However, these two groups of caregivers differed in the particular strategies they used to shape their children's behavior. Caregivers of mentally retarded children pointed to objects while caregivers of autistic children spent more time physically holding their children on task. Individual differences within the autistic sample indicated that caregivers regulated their children's behavior less and showed more mutual play and positive feedback to more communicatively able autistic children. These findings suggest that caregivers respond differentially to the specific deficiencies shown by their children. PMID- 2966190 TI - Rebound increase in plasma renin and vasopressin following graded infusions of atrial natriuretic peptide in man. AB - The effects of graded infusions of atrial natriuretic peptide (ANP) on hemodynamics, renal function, plasma renin activity (PRA) and plasma arginine vasopressin (AVP) were investigated in a two part study in 6 volunteers. Three 30 min iv infusions of either saline control or ANP at graded rates of 4, 10 and 40 pmol kg-1 min-1 were given. ANP infusions were associated with a significant increase in sodium clearance from 1.08 +/- 0.21 to 2.83 +/- 0.50 ml/min, an increase in hematocrit and a net fluid loss. Plasma AVP remained constant during ANP infusions but increased significantly afterwards when plasma ANP concentrations were falling rapidly, and this was accompanied by marked antidiuresis. PRA fell by 23% during the saline control infusions and by 50% during ANP infusions. Following cessation of ANP infusions there was a significant rebound increase in PRA. No changes were observed in blood pressure, heart rate, glomerular filtration rate or renal plasma flow. These results suggest an interaction in man between ANP and the hormones renin and AVP. PMID- 2966191 TI - Life table analysis and pathologic observations in male mice of a long-lived hybrid strain (Af X C57BL/6)F1. AB - We have utilized a long-lived (Af X C57BL/6)F1 hybrid strain of mice for a variety of aging studies. In this report we have characterized the life expectancy and pattern of spontaneous deaths in 202 mice, malignant and nonmalignant lesions in 64 male mice dying spontaneously, and organ weights and lesions in 39 male mice killed at selected ages. The maximum age observed was 41.5 months. The principal causes of death were malignant lymphoma and alveologenic neoplasms, which were present in 56.3% and 45.3%, respectively, of the mice dying spontaneously. A variety of other neoplastic and non-neoplastic lesions that are not infrequently seen in older mice were observed in these mice. Neoplasms seen in these mice that are rare in other mice included disseminated mast cell tumors in two mice and gastric adenocarcinoma in one mouse. In comparing the diseases observed in this hybrid strain with those reported for the parent strains, there was an incidence of malignant lymphoma similar to the C57BL/6 parent, an incidence of alveologenic neoplasms intermediate between the parent strains, and a markedly reduced incidence of amyloidosis. This study provides a detailed background of baseline hematologic and morphologic data in a long-lived hybrid of two commonly used strains of mice. PMID- 2966192 TI - Hepatitis B vaccine: persistence of antibody following immunization. AB - We examined the persistence of hepatitis B surface antibody following hepatitis B vaccination in a group of health care workers. Of the 187 vaccine recipients screened, 39 were found to be antibody negative. Life table analysis showed the proportion retaining antibody at one, two, three, and four years from the first dose of vaccine was 92%, 84%, 73%, and 55% respectively. The likelihood of retaining antibody was significantly decreased as the age of the vaccine recipient increased. These results indicate that a large proportion of vaccine recipients no longer had detectable antibody four years after receipt of the vaccine. It remains to be shown if such individuals who have lost antibody are susceptible to the development of clinical hepatitis. PMID- 2966193 TI - Increased platelet and vascular smooth muscle reactivity to low-dose adrenaline infusion in mild essential hypertension. AB - During low-dose adrenaline infusion, platelet count, platelet size, plasma beta thromboglobulin (BTG) and forearm vascular resistance (FVR) were measured in twelve 40-year-old men with mild, untreated hypertension. The average platelet count increased from 195 to 226 X 10(9)/l (P less than 0.001), platelet size from 7.31 to 7.53 X 10(-15)/l (P less than 0.01), BTG from 0.61 to 1.08 nmol/l (P less than 0.02) and FVR decreased from 97 to 58 (arbitrary units; P less than 0.001) during the infusion. The change in platelet count reflects splenic release of platelets, the change in plasma BTG reflects platelet release reaction, while the reduced FVR reflects vascular smooth muscle cell relaxation. In 11 normotensive men aged 40 years, platelet count increased from 187 to 201 X 10 g/l (P less than 0.01) during an equal low-dose adrenaline infusion. This increase in platelet count is significantly less than in the hypertensive group (P less than 0.01). There was statistically no significant change in platelet size, BTG or FVR in the normotensive group. Arterial adrenaline rose from 0.5 to 2.5 nmol/l in the hypertensive and from 0.5 to 2.4 nmol/l in the normotensive group. A third group of 12 normotensive men received saline infusion: neither platelet parameters nor FVR changed in this group. Thus, a small and equal dose of adrenaline elicited a greater increase in platelet count, an enhanced platelet release reaction and a more pronounced forearm vasodilation in hypertensive than in normotensive subjects. PMID- 2966194 TI - Monoclonal anti-idiotypic antibodies to an I-J interacting molecule inhibit suppression in an H-2 restricted way. AB - We have obtained 10 mAb from two independent fusions that are anti-idiotypic to the combining site of an anti-I-Jk antibody. These mAb block Ts cell function isn a genetically restricted manner in vitro and in vivo and recognize a determinant on macrophage membranes. In addition, they do not affect the I-Ek-restricted activation of a Th cell line specific for pigeon heart cytochrome c. We conclude that these mAb may recognize a molecule other than conventional I-Ek on cells interacting with Ts cells that is involved in mediating Ts activity. The molecule recognized may be a modified I-Ek molecule or a molecule not encoded by the genes encoding I-Ek. PMID- 2966195 TI - Cyclosporine blocks the activation of antigen-dependent cytotoxic T lymphocytes directly by an IL-2-independent mechanism. AB - The immunosuppressive drug Cyclosporin A (cyclosporine) inhibits the reactivation of quiescent Ag-dependent CTL in the presence of IL-2. Both proliferation and the regeneration of cytotoxicity are inhibited. The cytotoxic cells that are inhibited are Ag dependent for activation, whereas derived, Ag-independent, but still IL-2-dependent, cytotoxic cells are insensitive to cyclosporine. Cyclosporine also directly inhibits the effector phase of the cytotoxic cells, although not completely. The generation of primary CTL in mixed cultures was also blocked by cyclosporine in the presence of IL-2, in a time-dependent way that indicated that the sensitive time was early during the cultures. The CTL generated in primary cultures were significantly inhibited by cyclosporine in the assay, but this inhibition was less than for the cloned CTL lines. PMID- 2966196 TI - Modulation of class I and class II histocompatibility antigens on human T cell lines by IFN-gamma. AB - IFN-gamma is an immunomodulatory agent which is known to induce or enhance the expression of class II histocompatibility Ag (Ia Ag) on many lymphoid cells and cell lines of diverse origin. However, we have observed that IFN-gamma did not induce the expression of Ia Ag on Ia- human T cell lines. Neither did IFN-gamma enhance the expression of Ia Ag on Ia+ T cells. However, IFN-gamma was able to enhance the expression of class I histocompatibility Ag (HLA-A,B,C Ag) on a number of the T cell lines tested. Experiments with 125I-labeled IFN-gamma showed a relatively small degree of specific binding to these T cell lines. More extensive studies on two of the T cell lines demonstrated 1000 and 2600 IFN-gamma binding receptor sites/cell and binding affinities of 4.0 X 10(-10) M and 7.3 X 10(-10) M. Thus, although IFN-gamma can bind to human T cell lines and enhance class I histocompatibility Ag on these cells, IFN-gamma alone does not appear to regulate expression of class II histocompatibility Ag on T cell lines. PMID- 2966198 TI - Selective modulation of two human monocyte Fc receptors for IgG by immobilized immune complexes. AB - Two types of IgG FcR, FcRI and FcRII, are constitutively expressed by human monocytes. FcRI (identified by mAb 32.2) binds human (h) IgG, FcRII (identified by mAb IV.3) has a low affinity for hIgG but interacts strongly with murine (m) IgG1. These receptors can be assayed by using indicator E sensitized by hIgG (EA hIgG) or mIgG1 (EA-mIgG1), respectively. We further characterized these two FcR by modulation studies by using substrate-immobilized immune complexes containing rabbit IgG, goat IgG, or one of the mouse Ig classes or subclasses. After incubating monocytes in microtiter wells containing such immune complexes, binding of the two types of indicator red cells on the apical surface of the monocytes was quantitated using a photometric assay employing the pseudoperoxidase activity of E. No effect on the binding of sensitized E was observed after incubation of monocytes with immune complexes containing mouse IgE, IgA, or IgM, or F(ab')2 fragments of rabbit IgG. High concentrations of immune complexes containing IgG of mouse, rabbit, or goat, however, were able to induce a decrease in binding of both types of sensitized E, suggestive of modulation of both FcRI and FcRII. At lower concentrations of immune complexes, more selective patterns of modulation emerged. Under these conditions, immune complexes containing mIgG1 or mIgG2b, or, surprisingly, goat IgG induced a selective decrease in the binding of EA-mIgG1 (FcRII modulation), while immune complexes containing mIgG2a or rabbit IgG mainly affected the binding of EA-hIgG (FcRI modulation). By using anti-FcR mAb IV.3, it was confirmed that FcRII was modulated from the apical surface of monocytes after incubation on immune complex coated substrates. Selectivity of FcR-modulation was demonstrated by showing that under these conditions binding of anti-C receptor mAb, and several other anti monocyte mAb did not decrease. PMID- 2966197 TI - Modulation of human polymorphonuclear leukocyte IgG Fc receptors and Fc receptor mediated functions by IFN-gamma and glucocorticoids. AB - Human polymorphonuclear neutrophils (PMN) normally express two distinct types of IgG Fc gamma R, the 40-kDa Fc gamma R referred to as Fc gamma RII and the low affinity 50- to 70-kDa Fc gamma R designated Fc gamma RIII. A third type of Fc gamma R, the 72-kDa high affinity receptor known as Fc gamma RI, is also detectable on PMN that have been activated by IFN-gamma. Using mAb that discriminate among the three known types of Fc gamma R, we examined the effects of IFN-gamma and glucocorticoids on human PMN Fc gamma R expression. We also studied effects of IFN-gamma and the synthetic glucocorticoid dexamethasone (DEX) on antibody-dependent cytotoxicity (ADCC) of chicken erythrocytes and phagocytosis of IgG-coated ox RBC by human PMN. In 20 donors studied, we found that treatment of PMN with 400 U/ml IFN-gamma induced a 9- to 20-fold increase in the number of Fc gamma RI sites per cell, and DEX inhibited this induction of Fc gamma RI by 39 to 73%. Similarly, DEX significantly reduced the IFN-gamma stimulation of ADCC and phagocytosis. IFN-gamma had no effect on expression of Fc gamma RII or Fc gamma RIII. Fc gamma RI and Fc gamma RII expression was unaltered by 24 h of treatment with DEX alone, but Fc gamma RIII expression was sometimes increased by about 20% on PMN cultured with DEX. Nevertheless, we found a small but significant inhibition of ADCC and phagocytosis by 200 nM DEX. Our results indicate that Fc gamma RI plays a major but not exclusive role in the regulation of ADCC and phagocytosis by IFN-gamma and DEX. PMID- 2966199 TI - Peritoneal B cells respond to phorbol esters in the absence of co-mitogen. AB - B cells obtained by irrigation of the peritoneal cavity differ from splenic B cells in signaling requirements for the initiation of DNA synthesis. Splenic B cells are stimulated to enter S phase by phorbol esters in conjunction with a second signal provided by calcium ionophore; however, splenic B cells are not stimulated by phorbol ester alone. In contrast, peritoneal B cells from NZB and BALB/c mice were stimulated to incorporate tritiated thymidine by each of the phorbol esters, PMA and phorbol dibutyrate, acting alone. Stimulation of peritoneal B cells was apparent when cells were cultured at lower than usual cell densities, and responses were unaffected by coculture with splenic B cells. Responding cells adhered to plastic petri dishes coated with anti-mouse IgM antibody, but were not completely removed by treatment with anti-Ly-1.2 antibody plus C. These results indicate that phorbol esters constitute a complete signal that stimulates some peritoneal B cells to enter S phase. PMID- 2966200 TI - Mechanism of protection from graft-vs-host disease in murine mixed allogeneic chimeras. I. Development of a null cell population suppressive of cell-mediated lympholysis responses and derived from the syngeneic bone marrow component. AB - Splenocyte populations from whole body-irradiated recipients of mixed T cell depleted (TCD) syngeneic and allogeneic (complete H-2 disparity) bone marrow, or of TCD syngeneic marrow alone, contain cells with the ability to suppress the generation of cell-mediated lympholysis responses in vitro. This activity, which is present by 8 days after bone marrow transplantation and persists for several weeks, has been analyzed for possible veto-like or other specificity. Although reproducible patterns of suppression were observed, depending both on host strain and on the genetic combination of the response examined, the overall suppression in vitro most closely resembles that which has been ascribed to "natural suppressor" cells in other systems. The suppression appears to be mediated by a non-T cell, non-B cell, nonadherent, asialo GM1-negative population. Cold target inhibition and CTL activity of chimeric cells have been ruled out as factors contributing to the observed suppression. Significantly, in mixed chimeras, suppression was found to be mediated exclusively by cells which were syngeneic to the recipient in both recipient strains tested. The rapid development of this suppressive activity may explain the resistance to graft-vs-host disease conferred on whole body-irradiated mice by the addition of TCD syngeneic marrow to an allogeneic graft-vs-host disease-producing inoculum. PMID- 2966201 TI - CD4 molecules are associated with the antigen receptor complex on activated but not resting T cells. AB - To explore the relationship between CD4 and CD3/Ti on the T cell surface, we have studied a panel of Ag-specific Th cell lines and clones, as well as resting and mitogen-activated CD4+ cells. Our results show that exposure of Th cells to their specific antigenic stimuli, but not to irrelevant stimuli, induced the rapid disappearance of approximately 20 to 35% of CD3 and CD4 molecules. The modulation of these molecules was detected in less than 1 h, became maximal at 12 h, and recovered thereafter in parallel. Treatment of Th cells with anti-CD4 antibody prevented Ag-induced modulation of CD3, and treatment with anti-CD3 blocked modulation of CD4. In the absence of Ag, treatment of these cells with an antibody (WT-31) directed at a conformational determinant within CD3/Ti or with the combination of anti-CD3 antibody and goat anti-mouse Ig, also resulted in significant modulation of CD4. Similar treatment of PHA-activated CD4+ T cells with anti-CD3/Ti antibodies also induced CD4 modulation; however, the same antibodies failed to affect CD4 expression on fresh resting T cells. These results indicate that on activated, but not resting T cells, CD4 molecules can be physically associated with CD3/Ti. We postulate that this association is essential for efficient Th cell activation, and further that the ability of anti CD4 antibodies to inhibit helper functions is due to their prevention of CD4 CD3/Ti interaction on the T cell surface. PMID- 2966202 TI - Activation of B cells in vivo by a Fab/Fc fragment of a monoclonal anti-IgD antibody requires an interaction between the antibody fragment and a cellular IgG Fc receptor. AB - To investigate activation of B lymphocytes in vivo by an interaction between B cell surface Ig (sIg) and an anti-Ig antibody, we compared the abilities of a divalent IgG2b anti-IgD mAb, H delta a/1, and its univalent Fab/Fc fragment to enhance B cell sIa expression in vivo. The Fab/Fc fragment consists of a single Fab linked to Fc, and can interact with C and cellular Fc receptors. Although injection of BALB/c mice with either intact H delta a/1 or H delta a/1 Fab/Fc enhanced splenic B cell sIa expression, Ia expression was enhanced more by intact H delta a/1. Furthermore, injection of mice with 24G2, a mAb to the B cell and macrophage IgG2b Fc receptor, completely blocked the ability of 20 to 500 micrograms of H delta a/1 Fab/Fc to enhance B cell sIa expression, but had no effect on enhancement of B cell sIa expression by 100 micrograms of intact H delta a/1. This effect of 24G2 was mediated by its blocking of IgG2b receptor function rather than simply by its binding to B lymphocytes, since a mAb to the B cell IgE receptor did not interfere with the ability of H delta a/1 Fab/Fc to enhance B cell sIa expression. The different effects of 24G2 on B cell activation by intact H delta a/1 and H delta a/1 Fab/Fc were not a result of differences in the abilities of the intact antibody and its Fab/Fc fragment to activate B cells, since 24G2 did not interfere with the ability of AMS-15, a IgG2a anti-IgD mAb, to slightly increase B cell sIa expression. These observations indicate that a univalent ligand for B cell sIg can activate B lymphocytes in vivo through an IgGFc-IgGFc receptor-dependent interaction. PMID- 2966203 TI - Successful therapy of Theiler's virus-induced demyelination (DA strain) with monoclonal anti-Lyt-2 antibody. AB - The effects of therapy with mAb to T cell subsets were studied in mice with Theiler's murine encephalomyelitis virus-induced demyelination. mAb GK1.5 (directed at class II-restricted T cells) and mAb 2.43 (directed at class I restricted T cells) depleted the appropriate subset of T cells in lymph nodes, spleens, and peripheral blood for 6 to 8 wk after a single i.p. injection of 1 mg of purified mAb. Early treatment with mAb GK1.5 (days -1, 0, and +1 relative to virus injection) resulted in death, encephalitis, and increased demyelination in the majority of animals tested. Treatment with mAb 2.43 resulted in less meningeal inflammation and fewer demyelinating lesions in the spinal cord, irrespective of whether the mAb was given early or after demyelinating disease was established (days 15, 16, and 17). Beneficial response to mAb therapy did not correlate with titers of virus isolated from the central nervous system or serum. These results indicate an important role of class II-restricted T cells during early disease in preventing overwhelming encephalitis; class I-restricted T cells may be critical during demyelination. PMID- 2966204 TI - T-B collaboration in the in vitro anti-Sm autoantibody response of MRL/Mp-lpr/lpr mice. AB - Spontaneous production of autoantibodies to the Sm nuclear Ag is highly specific for SLE and the SLE-prone MRL mouse strains. Our previous studies have demonstrated that in vitro anti-Sm production in MRL/1pr mice requires the presence of T cells. In the present investigation, these T cells were found to express the L3T4+/Lyt-2- phenotype, unlike the aberrant L3T4-/Lyt-2-"double negative" 1pr T cells, and to utilize the L3T4 determinant in generating help for the anti-Sm response. The generation of anti-Sm did not require the presence of Sm-specific Th cells, as help could also be provided by T cells activated to an irrelevant Ag, or by nonspecific factors such as IL-2. There was no evidence for suppressor cell regulation of anti-Sm, even in animals negative for this specificity. These studies indicate that ongoing production of anti-Sm in MRL/1pr mice is dependent on the presence of T cells with a normal mature surface phenotype, and that these cells act in part through the elaboration of lymphokines. They further show that the anti-Sm status of individual MRL/1pr mice is not due to the action of suppressor cells. Because T cells appear to act primarily in a permissive fashion for the anti-Sm response, it is likely that events underlying the initial generation of Sm-specific B cell precursors are critical in determining whether an individual animal develops the Sm serologic specificity. Once these cells have arisen, clonal expansion of Sm-specific B cells may proceed in the presence of activated T cells or some of their products. PMID- 2966205 TI - Monoclonal antibodies to mouse complement receptor type 1 (CR1). Their use in a distribution study showing that mouse erythrocytes and platelets are CR1 negative. AB - mAb to murine C receptor type 1 (CR1) were produced and three of them were characterized. One antibody, designated as 8C12, immunoprecipitated a protein of 190,000 Mr from a detergent extract of surface-labeled spleen cells and stained spleen B but not T lymphocytes in fluorescent flow cytometry. It inhibited both CR1-mediated rosette formation and the cofactor activity of CR1 for factor I mediated cleavage of C3b, suggesting that it recognizes the ligand-binding site of CR1. The two other antibodies, designated as 7G6 and 7E9, recognized different epitopes from that recognized by 8C12, and they cross-reacted with a protein of 150,000 Mr that is present in a spleen extract. The distribution of CR1 in murine hemopoietic cells was studied by binding experiments with radiolabeled 8C12 and fluorescent flow cytometry. When CR1 was not detected by 8C12 alone, the two other antibodies were used in combination with 8C12 to confirm the negative results. Almost all B lymphocytes from the spleen, lymph nodes, and peripheral blood were CR1 positive. Most of the Thy-1-positive lymphocytes from these tissues were CR1 negative. Thymus lymphocytes were also CR1 negative. Peritoneal macrophages and chemotactic factor stimulated but not unstimulated peripheral blood granulocytes were CR1 positive. In contrast to human E, mouse E were CR1 negative. This pattern of distribution was consistent with previous results obtained by rosette assays. Although mouse platelets cause immune adherence hemagglutination with C3b-bearing SRBC, they are CR1 negative. Three other lines of evidence also indicated that platelets are CR1 negative. First, no band of CR1 was demonstrated by immunoprecipitation with 8C12 of an extract of surface labeled platelets. Second, 8C12, which inhibited rosette formation by lymphocytes, alone or in combination with 7G6 and 7E9, did not inhibit immune adherence between platelets and C3b-bearing SRBC. Third, polyclonal rabbit IgG prepared from anti-mouse CR1 antiserum did not inhibit immune adherence by platelets. These results strongly suggest that the C3b-binding factor(s) on mouse platelets is different from CR1 and that processing of C3b-bearing immune complexes in mouse blood may be mediated by a new and as yet unidentified C3b binding factor(s). PMID- 2966206 TI - Biochemical characterization of an Fc gamma receptor purified from human neutrophils. AB - The Fc receptor identified by mAb 3G8 (Fc gamma RIII) was isolated by mAb affinity chromatography from 0.5 to 2 x 10(10) neutrophils yielding 33 to 149 micrograms of protein. Iodination of the purified protein identified a polypeptide of broad electrophoretic mobility from Mr 47 to 70 kDa and occasionally a fainter polypeptide at 100 to 130 kDa, which may be dimerized receptor. Two-dimensional isoelectric focusing gel electrophoresis illustrated multiple diffuse polypeptides ranging from a pI of less than 4.7 to 6.5. Treatment of the purified receptor with neuraminidase shifted the mobility of these polypeptides to a more basic pI, ranging from 6 to 8, illustrating the presence of sialic acid residues on Fc gamma RIII. The glycoprotein nature of Fc gamma RIII was characterized by several criteria. The receptor bound to Con A Sepharose. Treatment of Fc gamma RIII with endoglycosidase H or F, which cleave high mannose and biantennary complex N-linked oligosaccharides, respectively, failed to alter the electrophoretic mobility of the Fc gamma R. Peptide N:glycosidase F, which cleaves all classes of N-linked oligosaccharides, reduced the Mr of Fc gamma RIII by 60% to reveal two poorly resolved polypeptides centered at Mr 25 kDa and ranging from Mr 16 to 28 kDa. Chemical deglycosylation with trifluoromethanesulfonic acid, which cleaves O- and N-linked oligosaccharides except for the asparagine-linked N-acetylglucosamine, reduced the Mr of Fc gamma RIII to 21 to 36 kDa. These results demonstrate that Fc gamma RIII is an acidic complex sialoglycoprotein and suggest that there may be 8 to 15 N-linked oligosaccharide chains on Fc gamma RIII. PMID- 2966207 TI - The biosynthesis and assembly of T cell receptor alpha- and beta-chains with the CD3 complex. AB - The biosynthesis, processing, and assembly of the TCR alpha- and beta-chains with each other and with the CD3 complex were investigated on both cell surface positive (TCR+CD3-) and negative (TCR-CD3-) cell lines. The results indicate that 1) in cell surface TCR-CD3- cell lines (MOLT 3, CCRF-CEM), TCR-beta, but not alpha-chains are present intracellularly. TCR-beta-CD3 complexes are readily found in these cell lines, but no evidence for final processing or cell surface expression of such incomplete TCR-CD3 complexes is observed. 2) In the cell surface TCR+CD3+ cell line HPB-ALL, both alpha- and beta-chains are present intracellularly. Whereas non-glycosylated forms of TCR-beta chain can be detected, only more mature forms of TCR alpha-chains are detected indicating that the alpha-chains are more rapidly glycosylated than the beta-chains. 3) The large majority of the intracellular alpha- and beta-chains is not disulfide linked and a small fraction of these is associated with CD3. 4) Only small amounts of the total intracellular TCR chains are found as CD3-associated disulfide-linked alpha beta-heterodimers. 5) Final processing of TCR chains for cell surface expression takes place after formation of these TCR-alpha beta-CD3 complexes. Thus, both the TCR alpha- and beta-chains are over-produced and only relatively small amounts of these chains form CD3-associated heterodimers that are processed for cell surface expression. Analogous results were obtained with a non-leukemic CTL clone. Based on these observations, a model for the biosynthesis and assembly of the TCR-CD3 complex is presented. PMID- 2966208 TI - The role of L3T4+ and Lyt-2+ T cells in the IgE response and immunity to Nippostrongylus brasiliensis. AB - The role of L3T4+ and Lyt-2+ T cells in protective immunity to Nippostrongylus brasiliensis (Nb) was studied in BALB/c mice that were depleted of either the L3T4+ or Lyt-2+ T cell population by injection with rat mAb specific for the appropriate determinant. Host responses to Nb infection including spontaneous elimination of adult worms, development of intestinal mucosal mast cell hyperplasia and the generation of a polyclonal IgE response were all completely blocked by 0.5 mg anti-L3T4 antibody administered simultaneously with Nb inoculation. However, administration of 0.5 mg of anti-Lyt-2 antibody at the same time and 7 days after inoculation with Nb had no effect on any of these responses. Injection of anti-L3T4 antibody as late as 9 days after Nb inoculation interfered with spontaneous cure of Nb infection and anti-L3T4 antibody injection 11 days after Nb inoculation inhibited serum IgE levels measured on day 13 by 50%. In addition, administration of anti-L3T4 antibody at the time of the peak serum IgE response, 13 days after Nb inoculation, accelerated the decline in serum IgE levels. Injection of previously Nb-infected mice with anti-L3T4 antibody at the time of a second Nb inoculation prevented the development of a secondary IgE response but did not affect immunity to Nb infection based on finding no adult worms in the intestines of these mice. These data indicate that 1) L3T4+ T cells are required for spontaneous cure of Nb infection, development of intestinal mucosal mast cell hyperplasia, and the generation and persistence of an IgE response during primary infection with Nb and 2) L3T4+ T cells are required for a considerable time after inoculation for optimal development of these responses. However, L3T4+ T cells are not required for all protective responses in immune mice. In contrast, our data indicate that considerable depletion of the Lyt-2+ T cell population has no significant effect on either worm expulsion or the generation of serum IgE responses. PMID- 2966209 TI - Anti-CD3 and phorbol myristate acetate regulation of MHC unrestricted T cell cytotoxicity. Lack of a requirement for CD3/T cell receptor complex expression during tumor cell lysis. AB - The effects of anti-CD3 mAb on MHC-unrestricted cytotoxic activity of NK depleted PHA-activated human T cells were examined. Anti-CD3 mAb had variable effects on killing of K562 or Daudi targets. Whereas lower concentrations of OKT3 often inhibited lysis of either target, higher concentrations (greater than 1 micrograms/ml) frequently increased K562 killing and always augmented Daudi lysis. However, lysis of the renal cell carcinoma, Cur, was consistently inhibited by OKT3 over a broad concentration range. Such variable effects were not related to differential regulation of heterogeneous subsets of effector cells, as similar patterns of OKT3-mediated modulation of tumor cell lysis by T cell clones was also observed. Another IgG2a anti-CD3 mAb, 64.1, and either F(ab')2 fragments of OKT3 or intact OKT3 in the presence of aggregated human Ig were found to inhibit lysis of Cur, K562, and Daudi targets consistently. Additional experiments were carried out to determine whether modulation of CD3 accounted for the inhibitory effects of the anti-CD3 mAb. PMA was noted to cause modulation of CD3 from the surface of PHA or alloantigen-activated T cells, and the combination of anti-CD3 and PMA caused even more marked modulation of CD3. Whereas preincubation with PMA and/or anti-CD3 decreased alloantigen-specific cytotoxic T cell function in relative proportion to the loss of CD3 expression, no consistent relationship between CD3 expression and the capacity of PHA activated T cells to kill Cur targets was noted. PMA alone caused no consistent alteration of Cur lysis. Moreover, in the presence of PMA, anti-CD3 mAb caused no significant inhibitory effect on Cur lysis, in spite of increased modulation and in some cases virtual total loss of surface CD3 expression. These findings indicate that when FcR interactions are prevented, anti-CD3 mAb consistently inhibit MHC-unrestricted cytotoxicity by PHA-activated T cells. Despite this, the data support the conclusion that CD3/TCR complex interactions with target cells are not required for either target cell recognition or triggering of lysis by MHC unrestricted cytotoxic T cells. PMID- 2966210 TI - High-dose irradiated splenic stimulator cells show no endogenous interleukin-2 production but stimulate clonally developing helper T cells to produce interleukin-2. AB - The effect of various physical or chemical treatments of splenic stimulator cells on their endogenous, mitogen-inducible IL-2 production and on their ability to induce IL-2 production in clonally developing helper T lymphocytes was investigated. While most methods (T cell depletion by monoclonal antibodies plus complement, glutaraldehyde fixation, heat inactivation and high-dose irradiation) effectively suppressed the endogenous IL-2 production of splenic stimulator cells, only T cell depletion and high-dose (6000-10,000 R) irradiation sustained their stimulatory capacity. High-dose irradiated stimulator cells induced high numbers of clonally developing helper T lymphocytes to secrete IL-2. Moreover, this induction was found to be antigen-specific. Hence, high-dose irradiation is a simple, rapid and reliable method for the treatment of stimulator cells, especially when large numbers of cultures are to be screened. PMID- 2966211 TI - Characterization of T lymphocyte responses during primary infection with human immunodeficiency virus. AB - The present study was undertaken to determine T cell response to primary human immunodeficiency virus (HIV) infection. No significant difference in T cell subsets was found between subjects who later seroconverted and a group of controls. Six subjects had multiple enumerations of T cell subsets done at the time of seroconversion. Initially, total lymphocyte and T cell subset counts were reduced. An inversion of the CD4+:CD8+ ratio due to a rise in the level of CD8+ cells was found later, followed by an appreciable increase in the number of CD8+ cells and further inversion of the CD4+:CD8+ ratio. Finally, the CD8+ cell count returned toward normal but remained higher than the CD4+ cell count; the inverted ratio was maintained. Lymphocyte hyporesponsiveness to mitogens and antigens was found during the seroconversion illness in one subject. In three of five subjects for whom data were available, an increase in the absolute number of CD8+ cells followed a decrease in the serum HIV antigen level. PMID- 2966212 TI - Glasgow Coma Scale scores in the patient post cardiopulmonary resuscitation. AB - This study was conducted to examine the difference in pre- and post-resuscitation Glasgow Coma Scale (GCS) scores in patients who suffered a cardiopulmonary arrest. Patients who arrested in an acute hospital setting were compared in terms of Glasgow Coma Scale scores with patients who arrested outside an acute hospital setting. The subjects for this study were 39 patients who arrested in the hospital and 16 patients who arrested outside the hospital setting. Data were collected over a period of one year using chart review. The data for this study included: GCS scores at one, two, four, twelve, and twenty-four hours post arrest; age of the subjects; sex; site of arrest; and etiology of arrest. Results indicated that hospitalized arrest subjects had significantly higher GCS scores than out-of-hospital arrest subjects. Results further indicated that younger patients and those with myocardial infarctions had significantly higher GCS scores post-arrest. This study has implications for serial neurological assessment of the patient post-arrest. PMID- 2966213 TI - Neuromuscular respiratory failure: what the nurse knows may make the difference. AB - The incidence of pulmonary compromise in neuromuscular disease is a well established phenomenon. Classic symptoms of impending respiratory failure may be absent or nebulous however, in the neuromuscular patient. It is for this reason, that the neuroscience nurse should develop expertise in both the neuro and pulmonary sciences. The focus of this article will be on two neuromuscular patient populations at high risk of respiratory failure--myasthenics and those with Guillain-Barre Syndrome. PMID- 2966215 TI - Are we creating our own ethical dilemmas? PMID- 2966216 TI - Continuing education in neuroscience nursing. AB - Since its inception in 1968 the American Association of Neuroscience Nurses (AANN) has devoted the majority of its energy and resources to establishing and maintaining excellence in continuing education (CE). Building and sustaining a program for CE is both challenging and rewarding. AANN, accredited as a provider and approver of CE in nursing by the American Nurses' Association, has awarded hundreds of contact hours to individuals and AANN chapters. It now has the authority to approve CE for any organization or individual meeting specific criteria. In order to promote ongoing growth of its CE program, and thus the knowledge of nurses caring for neuroscience patients, AANN encourages groups and individuals to provide CE and offers this article as assistance. The article defines terms, addresses components of development of CE and offers the opportunity to earn contact hours. PMID- 2966214 TI - A method for quantifying the area of closed pressure sores by sinography and digitometry. AB - Neurologically impaired and elderly patients who remain in the same position for prolonged periods risk developing pressure sores. The most insidious of these are closed pressure sores. Conservative treatment of closed pressure sores frequently is the only treatment option. However, no systematic method to assess the effectiveness of conservative treatment modalities exists. The purposes of this study were to develop a method for quantifying the area of closed pressure sores by means of sinography and digitometry and to determine the utility of this method for monitoring healing. A convenience sample of 40 veterans with closed pressure sores was selected. Four individuals independently traced 116 regions from 30 sinograms of closed pressure sores from a subsample of ten patients. A fifth individual calculated areas of closed pressure sores in units, using digitometry. Findings and nursing implications will be presented. PMID- 2966217 TI - Validation of defining characteristics: retrospective design. AB - The American Association of Neuroscience Nurses (AANN) has accepted the concept of nursing diagnosis as one component of a general nursing framework which is designed to direct and evaluate the care of individuals with neuroscience dysfunction. Continuing development of nursing diagnosis as a conceptual basis for neuroscience practice is dependent upon several factors, one of which is research-based identification and validation of defining characteristics associated with nursing diagnoses commonly used in the care of patients with neurologic dysfunction. Defining characteristics are observable signs and symptoms present in the person with the problem. Identification and validation of defining characteristics are essential to the continued development and use of nursing diagnosis in nursing practice. Without clear, specific defining characteristics, neuroscience nurses are unable to effectively assess patients for the signs and symptoms necessary to formulate nursing diagnoses, monitor patient status or evaluate nursing interventions. One way to validate defining characteristics is the use of retrospective design. PMID- 2966218 TI - Pathology, treatment and management of posterior fossa brain tumors in childhood. AB - Brain tumors are the second most common childhood malignancy. Between 1975 and 1985, 462 newly diagnosed patients were treated at the Children's Hospital of Philadelphia; 207 (45%) tumors arose in the posterior fossa and 255 (55%) appeared supratentorially. A wide variety of histological subtypes were seen, each requiring tumor-specific treatment approaches. These included primitive neuroectodermal tumor (n = 86, 19%), astrocytoma (n = 135, 30%), brainstem glioma (n = 47, 10%), anaplastic astrocytoma (n = 32, 7%), and ependymoma (n = 30, 6%). Because of advances in diagnostic abilities, surgery, radiotherapy, and chemotherapy, between 60% and 70% of these patients are alive today. Diagnostic tools such as computed tomography and magnetic resonance imaging allow for better perioperative management and follow-up, while the operating microscope, CO2 laser, cavitron ultrasonic aspirator and neurosurgical microinstrumentation allow for more extensive and safer surgery. Disease specific treatment protocols, utilizing radiotherapy and adjuvant chemotherapy, have made survival common in tumors such as medulloblastoma. As survival rates increase, cognitive, endocrinologic and psychologic sequelae become increasingly important. The optimal management of children with brain tumors demands a multidisciplinary approach, best facilitated by a neuro-oncology team composed of multiple subspecialists. This article addresses incidence, classification and histology, clinical presentation, diagnosis, pre-, intra- and postoperative management, long term effects and the team approach in posterior fossa tumors in childhood. Management of specific tumor types is included as well. PMID- 2966219 TI - Infections related to intracranial pressure monitoring. AB - Infection related to the placement of intracranial pressure (ICP) monitoring systems has been examined closely by practitioners. Review of the literature identifies many factors contributing to infection. In this two-year retrospective study of 52 patients who underwent ICP monitoring, 12 variables were studied: diagnosis, type of monitor, date and place of insertion, duration of monitoring, drainage, frequency of system change or irrigation and personnel involved, dressing changes, range of patient temperatures, culture results, concurrent infections and antibiotic therapy. Practices that minimized infection rates in our patient population are identified as well as implications for nursing practice and quality assurance measures for ICP monitoring. PMID- 2966221 TI - [Simple fixation of the vaginal stump to the abdominal wall in uterine prolapse]. PMID- 2966220 TI - Perigraft-reaction after implantation of vascular prostheses. Pathogenesis, clinical picture and treatment. AB - This report is based on 18 perigraft reactions in 2237 implanted vascular prostheses (1970-1985). The incidence of this complication is 6.8-9.5/1000 both for Dacron-double-velour and expanded PTFE-prostheses. The clinical picture is characterized by localized or extended fluid accumulation around the artificial graft (perigraft cysts or "swimming prosthesis"). Including 271 reports from the literature, 75% of vascular prostheses in extraanatomical positions were affected. The time interval between vascular substitution and clinical manifestation is on average 23.5 months. The pathogenesis of this aseptic late complication is still unclear. Several disposing factors are under discussion: (a) physico-chemical irritation of the surrounding tissue by the fabric itself (release of monomers, emulgators etc.), (b) mechanical trauma (continuous movement of the graft in the tissue bed, i.e. insufficient tissue fixation resulting in gaps and exudation of fluid round the prosthesis), (c) latent or manifest renal insufficiency. The partial or total removal of the affected prosthesis and its substitution by another type of vascular prosthesis offers the best chance of definitive cure. Repeated puncture of perigraft cysts produces frequent recurrencies and the danger of secondary infections. Therefore only an exchange of the graft can be recommended. PMID- 2966223 TI - [Combined therapy with bromocriptine, clomiphene and gonadotropin for polycystic ovary syndrome patients failing to respond to clomiphene alone]. AB - The purpose of this study is to investigate the ovulatory effect of the combined therapy with bromocriptine (Brc), clomiphene (Cl) and HMG. We investigated 25 patients with polycystic ovary syndrome (PCOS) who had anovulatory normoprolactinemia and failed to respond to Cl therapy alone. The results were as follows: 1) The ovulation rate with Method I(Brc/Cl) was 64.0% (16/25) in 25 cases, and 59.8% in 122 cycles. 2) Resting levels of serum dehydroepiandrosterone sulfate in the effective group with Method I were significantly lower than those in the non-effective group. 3) With Method I, serum prolactin (PRL), LH and testosterone were significantly decreased, while estradiol and progesterone concentrations were significantly increased in the effective group. However, there were no significant changes in the hormone levels except PRL in the non effective group. 4) Out of the 9 nonrespondent Method I cases, 5 additional cases ovulated with both Method II(Brc/Cl+ a small dose of HMG) and Method III(Brc/Cl+ an increased dose of HMG-HCG). The total ovulatory rate was 84.0% (21/25) in 25 cases and 59.4% (85/143) in 143 cycles. 5) The total pregnancy rate was 43.8% (7/16). Six cases were normal pregnancies, one case was a twin pregnancy and there was no case of abortion. Our results suggested that the combined therapy with Brc, Cl and HMG had almost the same therapeutic effect as the HMG-HCG therapy in severe cases of PCOS. PMID- 2966222 TI - [A case of stromal endometriosis (endolymphatic stromal myosis) in the effective treatment with progestogen]. PMID- 2966225 TI - Breast reconstruction. PMID- 2966224 TI - Protein S deficiency in sickle cell anemia. AB - To investigate the status of the protein C-protein S anticoagulant pathway in sickle cell disease, we measured protein C, total and free protein S, and C4b binding protein levels in 20 subjects with sickle cell disease (Hb SS or SC). Mean total and free protein S levels were both significantly lower in subjects with sickle cell disease than in normal individuals, but greater reductions were observed for free S. The free protein S level was below the mean -2 SD for normal subjects in 12 subjects with sickle cell disease; the total protein S level was below this level in three subjects. Mean C4b-binding protein levels were normal in subjects with sickle cell disease, both during painful crisis and in the steady state, and no correlation was observed between the levels of C4b-binding protein and free protein S, suggesting that the low free protein S level was not caused by increased levels of C4b-binding protein. Crossed immunoelectrophoresis of plasma samples from eight subjects with sickle cell disease showed marked reductions in free protein S, with normal levels of protein S bound to C4b binding protein. In contrast to the protein S level, mean protein C activity was normal in subjects with sickle cell disease, both during painful crisis and in the steady state. However, the protein C level was below the mean -2 SD for normal subjects on at least one occasion in four subjects with sickle cell disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966226 TI - Dentists' professional and ethical responsibilities for HIV-positive patients and patients with AIDS. Federation Dentaire Internationale. PMID- 2966227 TI - Infectious disease control practices of Massachusetts dentists. PMID- 2966228 TI - An investigation of young infants' perceptual representations of speech sounds. AB - The present study examined the ability of newborns and 2-month-olds to detect phonetic differences between syllables. By relying on the modified high-amplitude sucking procedure, which did not permit the infants to use a simple same different response, the present experiments tapped the perceptual representations of the speech sounds. Infants as young as a few days old displayed some capacity to represent differences in a set of syllables varying in their phonetic composition, although there was no convincing evidence that their representations were structured in terms of phonetic segments. Finally, evidence of developmental changes in speech processing were noted for the first time with infants in this age range. The change noted was a tendency from global toward more specific representations on the part of the older infants. PMID- 2966229 TI - A multiplicative model of the utility of survival duration and health quality. AB - Survival duration and health quality are fundamentally important aspects of health. A utility model for survival duration and health quality is a model of the subjective value of these attributes. We investigate the hypothesis that the utility (subjective value) of survival duration and health quality is determined by a multiplicative model. According to this model, there are separate subjective scales for the utility of survival duration and health quality. If F(Y) equals the utility of surviving Y years, and G(Q) equals the utility of living in health state Q, then the multiplicative model proposes that F(Y)G(Q) equals the utility of surviving Y years in health state Q. This model provides a simple explanation for several intuitively compelling relationships. First, the distinction between better-than-death and worse-than-death health states corresponds to the assignment of positive or negative utilities to different health states. Second, a zero duration of survival removes any reason to prefer one health state over any other, just as multiplying the utility of health quality by zero eliminates differences between the utilities of different health states. Third, the subjective difference between Y years in pain and Y years free from pain increases as Y increases as if the difference in utility between pain and no pain were being multiplied by the utility of surviving Y years. A critical prediction of the multiplicative model is the hypothesis that preferences between gambles for health outcomes satisfy a property called utility independence. Individual analyses revealed that most subjects satisfy utility independence, thereby supporting the multiplicative utility model. Some subjects appear to violate a fundamental assumption of utility theory: They appear to violate the assumption that a single utility scale represents both the ordinal preference relations between certain outcomes and the subjective averaging that underlies the utility of gambles. The violation is inferred from an inconsistency between preferences for multiattribute outcomes when they are viewed as certain outcomes and when they are viewed as the outcomes of gambles. PMID- 2966230 TI - Pragmatics of measuring recognition memory: applications to dementia and amnesia. AB - This article has two purposes. The first is to describe four theoretical models of yes-no recognition memory and present their associated measures of discrimination and response bias. These models are then applied to a set of data from normal subjects to determine which pairs of discrimination and bias indices show independence between discrimination and bias. The following models demonstrated independence: a two-high-threshold model, a signal detection model with normal distributions using d' and C (rather than beta), and a signal detection model with logistic distributions and a bias measure analogous to C. C is defined as the distance of criterion from the intersection of the two underlying distributions. The second purpose is to use the indices from the acceptable models to characterize recognition memory deficits in dementia and amnesia. Young normal subjects, Alzheimer's disease patients, and parkinsonian dementia patients were tested with picture recognition tasks with repeated study test trials. Huntington's disease patients, mixed etiology amnesics, and age matched normals were tested by Butters, Wolfe, Martone, Granholm, and Cermak (1985) using the same paradigm with word stimuli. Demented and amnesic patients produced distinctly different patterns of abnormal memory performance. Both groups of demented patients showed poor discrimination and abnormally liberal response bias for words (Huntington's disease) and pictures (Alzheimer's disease and parkinsonian dementia), whereas the amnesic patients showed the worst discrimination but normal response bias for words. Although both signal detection theory and two-high-threshold discrimination parameters showed identical results, the bias measure from the two-high-threshold model was more sensitive to change than the bias measure (C) from signal detection theory. Three major points are emphasized. First, any index of recognition memory performance assumes an underlying model. Second, even acceptable models can lead to different conclusions about patterns of learning and forgetting. Third, efforts to characterize and ameliorate abnormal memory should address both discrimination and bias deficits. PMID- 2966231 TI - Problem structure and the use of base-rate information from experience. AB - This article is concerned with the use of base-rate information that is derived from experience in classifying examples of a category. The basic task involved simulated medical decision making in which participants learned to diagnose hypothetical diseases on the basis of symptom information. Alternative diseases differed in their relative frequency or base rates of occurrence. In five experiments initial learning was followed by a series of transfer tests designed to index the use of base-rate information. On these tests, patterns of symptoms were presented that suggested more than one disease and were therefore ambiguous. The alternative or candidate diseases on such tests could differ in their relative frequency of occurrence during learning. For example, a symptom might be presented that had appeared with both a relatively common and a relatively rare disease. If participants are using base-rate information appropriately (according to Bayes' theorem), then they should be more likely to predict that the common disease is present than that the rare disease is present on such ambiguous tests. Current classification models differ in their predictions concerning the use of base-rate information. For example, most prototype models imply an insensitivity to base-rate information, whereas many exemplar-based classification models predict appropriate use of base-rate information. The results reveal a consistent but complex pattern. Depending on the category structure and the nature of the ambiguous tests, participants use base-rate information appropriately, ignore base-rate information, or use base-rate information inappropriately (predict that the rare disease is more likely to be present). To our knowledge, no current categorization model predicts this pattern of results. To account for these results, a new model is described incorporating the ideas of property or symptom competition and context-sensitive retrieval. PMID- 2966232 TI - The role of context in interpreting facial expression: comment on Russell and Fehr (1987). AB - In their article, "Relativity in the Perception of Emotion in Facial Expressions," Russell and Fehr (1987) argued that context is the principal determinant in interpreting facial expressions of emotion. They questioned the biological bases for emotion suggested by Darwin and supported by many cross cultural studies. We suggest that their results occurred because the target faces they used were emotionally neutral or ambiguous. We also argue that their findings can be interpreted as supporting the communicative importance of the face. PMID- 2966233 TI - Is it smart to play the horses? Comment on "A day at the races: a study of IQ, expertise, and cognitive complexity" (Ceci & Liker, 1986). AB - Ceci and Liker (1986b) presented data that they contended shows two things: (a) Handicapping harness races is a cognitively complex undertaking that can be captured by a multiple regression model, and (b) neither overall skill at handicapping nor the complexity of the mental model used is related to standard measures of intelligence. The first contention is not at issue. But the second contention, that handicapping performance is unrelated to IQ, is not supported by the data presented; in fact, the opposite conclusion seems more likely. The purpose of this comment is to point out errors frequently made in individual differences research concerning population definition, sample selection, dependent and independent variable reliability and validity and interpretation of results. PMID- 2966234 TI - The relationship between glycosyltransferase production and membrane fatty acid composition of Streptococcus sanguis NCTC 7865 grown in the presence of protonmotive force inhibitors. AB - The fatty acid composition of Streptococcus sanguis NCTC 7865 was not altered by changing the cation composition (Na+/K+) of the growth medium; glucosyltransferase (GTF; EC 2.4.1.5) also remained constant. In contrast, fructosyltransferase (FTF-S; EC 2.4.1.10) production was reduced by at least 50% in medium with a high Na+ concentration. Growth in the presence of ionophores (gramicidin, nigericin or valinomycin) resulted in an increased proportion of saturated fatty acids, principally octadecanoic acid (C18:0), while the proportion of unsaturated fatty acids, predominantly octadecenoic (C18:1) and hexadecenoic (C16:1) acids, decreased. GTF-S production was reduced in the presence of ionophores whereas FTF-S production was completely abolished. Tween 80 significantly increased both GTF-S production and the proportion of unsaturated fatty acids in the cytoplasmic membrane; FTF-S production was unaltered by Tween 80. The production of GTF-S was inversely proportional to the C18:0:C18:1 fatty acid ratio of the cytoplasmic membrane. It was concluded that FTF-S production is directly influenced by protonmotive force (pmf), whereas GTF S production is affected more by the physical properties of the cytoplasmic membrane, in particular its fatty acid composition. However, as perturbations in pmf generation can lead to variations in membrane fatty acid composition it can be argued that pmf indirectly influences GTF production by changing the saturated:unsaturated or C18:0:C18:1 fatty acid ratio of the cytoplasmic membrane. PMID- 2966235 TI - Selective alteration in blood-brain barrier and insulin transport in iron deficient rats. AB - Nutritional iron deficiency induced in rats causes a significant reduction in level of brain nonheme iron and is accompanied by selective reduction of dopamine D2 receptor Bmax. Our previous studies have clearly demonstrated that these alterations can be restored to normal by supplementation with ferrous sulfate; however, neither brain nonheme iron level nor dopamine D2 receptor Bmax can be increased beyond control values even after long-term iron therapy. The possibility that iron deficiency can induce the breakdown of the blood-brain barrier (BBB) was examined. A 70 and 100% increase in brain uptake index (BUI) for L-glucose and insulin, respectively, were noted in iron-deficient rats. However, the BUI for valine was decreased by 40%, and those for L-norepinephrine and glycine were unchanged. In addition, it was demonstrated that in normal rats insulin is transported into the brain. The data show that iron deficiency selectively affects the integrity of the BBB for insulin, glucose, and valine transport. Whether the effect of iron deficiency on the BBB is at the level of the capillary endothelial cell tight junction is not yet known. However, this study has shown that an important nutritional disorder (iron-deficiency anemia) has a profound effect on the BBB and brain function. PMID- 2966236 TI - Effects of polyhydric and monohydric compounds on the stability of type I receptors for adrenal steroids in brain cytosol. AB - We have shown previously that unoccupied type I receptors for adrenal steroids in brain cytosol lose their capacity to bind [3H]aldosterone ([3H]ALDO) in a time- and temperature-dependent manner. Based on reports that sugars and polyvalent alcohols are capable of stabilizing a variety of globular proteins, we attempted in the present study to stabilize type I receptors by including polyhydric compounds in our brain cytosol preparations. However, contrary to expectations, adjusting cytosol to a 10% (g/dl) concentration of ethylene glycol, glycerol, erythritol, xylitol, ribitol, or sorbitol failed to stabilize these receptors at 0 degree C and in fact produced a slight reduction in [3H]ALDO binding capacity. The magnitude of this reduction was greater when cytosol was incubated for 2 h at 22 degrees C prior to incubation with [3H]ALDO. In contrast to these results, when brain cytosol was adjusted to a 10% (g/dl) concentration of the monohydric compound, ethanol, a significant increase in [3H]ALDO binding to type I receptors was found. Under identical conditions, methanol and propanol failed to have a significant effect on the binding capacity of these receptors. When cytosol was aged for 2 h at 22 degrees C, all three of these monohydric compounds produced a marked loss in the [3H]ALDO binding capacity of type I receptors. An investigation of various doses of ethanol at 0 degree C on the subsequent binding of [3H]ALDO yielded an inverse U-shaped curve with 10% ethanol producing the highest level of specific binding, as reflected by an increase in maximal binding in Scatchard plots, and 40% ethanol producing a complete loss in type I receptor binding capacity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966237 TI - Hexosaminidase A deficiency presenting as juvenile progressive dystonia. PMID- 2966240 TI - Management of von Willebrand's disease with desmopressin. PMID- 2966239 TI - Dermatitis from plastic tote boxes impregnated with an antistatic agent. AB - An outbreak of dermatitis occurred among employees of a microelectronics firm. In a cross-sectional epidemiologic investigation, we found that dermatitis of the hands or arms had occurred among 14 of 29 (48.3%) employees of the incoming inspection department where plastic tote boxes recently purchased from one manufacturer had been used, compared to only one case among 17 (5.9%) employees in another department which had not used these boxes. Affected workers could detect an oily film on the surfaces of these new boxes, but not on older ones. We identified the oily film to be a surface accumulation of bis-hydroxyethyl-tallow amine (BHETA), an antistatic agent with which the tote boxes had been impregnated. Subsequent toxicologic investigation established that BHETA could provoke both follicular and nonfollicular irritant dermatitis, and was also a potential skin sensitizer. Antistatic agents should be considered as potential causes of dermatitis among employees who handle electrical parts transported in plastic boxes, particularly when affected employees can detect an oily film on the box surfaces. PMID- 2966241 TI - House Bill 2067 and HIV-positive patients: legal implications for dental practice. PMID- 2966238 TI - Modulation by RU 41740 of oxidative metabolism and migration of rat polymorphonuclear leukocytes collected after induction of immune and nonimmune inflammations. AB - The activity of an immunomodulating agent, RU 41740, on the behavior of polymorphonuclear leukocytes (PMNs) elicited by two models of inflammation was studied. We observed the effect produced on migration and oxidative metabolism after incubation in vitro, in various drug concentrations, or administration in vivo. A stimulation of oxidative metabolism and an inhibition of chemotaxis was observed with the two types of PMNs. However Arthus-elicited PMNs were less reactive. This might be related to a nonspecific deactivation of the cells after induction of such an immune inflammatory reaction. This evidenced that PMN reactivity greatly varies with the physiopathological state of the cells. PMID- 2966243 TI - [Differences in hemodynamic responses to dynamic exercise between borderline and sustained hypertension]. AB - To observe the effects of anatomical distribution of left ventricular hypertrophy (LVH) and the differences in blood pressure on cardiac function, borderline hypertensives (group A) and mild or moderate sustained hypertensives (group B) were studied at rest and during dynamic exercise and their results were compared with those of normal controls (group C). LVH was estimated by M-mode echocardiography, and cardiac function was assessed by radionuclide (RI) first pass and multigate methods. In each subjects, 0.5 watt/kg and 1.0 watt/kg loads of ergometer exercise were administered for 16 min in the supine position. The main results were as follows: 1. In borderline hypertension, there was significant hypertrophy of the interventricular septum, and the left ventricular mass index was greater than that in normal controls. 2. The cardiac index in borderline hypertension did not differ from that of normal controls, but that of sustained hypertension was significantly reduced. 3. Significant positive correlations were observed between blood pressure and the left ventricular mass index at rest and during exercise. 4. No difference in left ventricular ejection fraction (LVEF) at rest was observed among the three groups, but that in sustained hypertension during exercise was markedly less than that of the other groups. LVEF at 1.0 watt/kg load and delta LVEF correlated inversely with the left ventricular mass index. 5. Left ventricular end-systolic volume determined by an RI count method decreased significantly during exercise in normals and borderline hypertension, while that in sustained hypertension was not reduced during exercise. Accordingly, cardiac function in the latter group was judged to be impaired. 6. In borderline hypertension, regional LVEF in the septal portion was slightly reduced at rest and during exercise. This reduction might have been caused by an imbalance of hypertrophy between the septal and free wall portions. In sustained hypertension, regional septal LVEF was maintained at rest, but was markedly less during exercise as compared with the other groups. Regional LVEF in the free wall portion was similar in degree to that of LVEF at rest and during exercise in the three groups. 7. Though the peak filling rate (PFR) in borderline hypertension did not differ from that in controls, that of sustained hypertension was significantly reduced at rest and during exercise. Significant positive correlations were obtained between PFR and LVEF at rest and during exercise. In summary, elevated blood pressure at rest and during exercise was thought to be the most important causative factor of LVH in hypertension.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966244 TI - Uterine and umbilical artery Doppler velocimetry in postterm pregnancy. AB - Uterine and umbilical artery Doppler velocimetry was carried out on 149 women whose pregnancies went to 41 weeks or beyond. Flow velocity was not altered even in the presence of other signs suggestive of fetal compromise. It appears that the postdate syndrome is not associated with significant alterations in the maternal or umbilical blood flow prior to the onset of labor. PMID- 2966242 TI - [Evaluation of coronary reperfusion for acute myocardial infarction by emission CT using technetium-99m pyrophosphate]. AB - Twelve patients with acute transmural myocardial infarction (AMI) were treated with percutaneous transluminal coronary angioplasty (PTCA) following intracoronary thrombolysis using urokinase, and underwent technetium-99m stannous pyrophosphate (Tc-99m-PPi) imaging 9.2 +/- 2.1 hours after the onset of chest pain. The imaging was performed with emission computed tomography (ECT). Compared to planar imaging, this allowed more accurate detection of small myocardial infarcts and accurate measurements of infarcts irrespective of their location was also made. Early Tc-99m-PPi images were obtained to test the hypothesis that an early, abnormal Tc-99m-PPi image suggest successful reperfusion. The results were presented for two groups of patients: three with unsuccessful reperfusion (Group A) and nine with successful reperfusion (Group B). Eight of the nine patients with successful reperfusion had positive acute Tc-99m-PPi images. On the contrary, all the three patients for whom reperfusion failed had negative acute Tc-99m-PPi images. We also examined the feasibility of estimating infarct size using positive Tc-99m-PPi images in patients with successful reperfusion during the early phase of AMI. The Tc-99m-PPi uptake score (Tc-US) was used to measure infarct size in this study. Areas of increased Tc-99m-PPi uptake within myocardial infarcts were threshold at 60% of the peak activity. The Tc-US of each patient was obtained to sum the scores of all myocardial segments using a scoring system with a maximum score of 108. Using this method, Tc-US ranged from 2 to 39. The correlation of Tc-US with the peak serum creatine kinase level was significant (r = 0.91).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966245 TI - Dopamine D-2 receptor imaging radiopharmaceuticals: synthesis, radiolabeling, and in vitro binding of (R)-(+)- and (S)-(-)-3-iodo-2-hydroxy-6-methoxy-N- [(1-ethyl 2-pyrrolidinyl)methyl]benzamide. AB - In developing central nervous system (CNS) dopamine D-2 receptor imaging agents, enantiomers, R-(+) and S-(-) isomers, of 3-[125I]iodo-2-hydroxy-6-methoxy-N-[(1 ethyl-2- pyrrolidinyl)methyl]benzamide, [125I]IBZM, were synthesized, and their in vitro binding characteristics were evaluated in rat striatum tissue preparation. The (S)-(-)-[125I]IBZM showed high specific dopamine D-2 receptor binding (Kd = 0.43 nM, Bmax = 0.48 pmol/mg of protein). Competition data of various ligands for IBZM binding displayed the following rank order of potency: spiperone greater than (S)-(-)-IBZM greater than (+)-butaclamol much greater than (R)-(+)-IBZM greater than (S)-(-)-BZM greater than dopamine greater than ketanserin greater than SCH23390 much greater than propanolol. The results indicate that [125I]IBZM binds specifically to the dopamine D-2-receptor with stereospecificity. The [123I]IBZM is potentially useful as an imaging agent for the investigation of dopamine D-2 receptors in humans. PMID- 2966246 TI - Homoallylic amines related to zimeldine. A comparative study on neuronal serotonin and norepinephrine reuptake based on conformational analysis. AB - A number of tertiary and secondary homoallylic amines, i.e. (Z)- and (E)-4-(4 bromophenyl)-4-(3-pyridyl)-3-buten-1-ylamines, were synthesized in diastereomerically pure forms. The compounds were evaluated as neuronal norepinephrine (NE) and serotonin (5-HT) uptake inhibitors under in vitro and ex vivo conditions and compared with the tricyclics amitriptyline and nortriptyline having homoallylic side chains and with the corresponding diastereomers in the zimeldine series having allylic side chains. The Z isomers of the new homoallylic derivatives (3Z, 4Z) were specific 5-HT uptake inhibitors in analogy with the corresponding allylic derivatives zimeldine (1Z) and norzimeldine (2Z). Likewise, the selectivity profile of the homoallylic (3E, 4E) and the allylic (1E, 2E) derivatives was comparable. In general, the homoallylic compounds were less potent inhibitors than their allylic counterparts. The similarities and discrepancies were evaluated in terms of conformational preferences determined by CAMSEQ molecular mechanics calculations. Homonorzimeldine (4Z) can accommodate energetically favored, but less populated, conformations having amino nitrogen atom to aromatic ring center distances comparable to those in norzimeldine. These facts correlate to retained 5-HT selectivity but diminished potency of 4Z compared to 2Z. PMID- 2966247 TI - Synthesis and pharmacological evaluation of 5,6-exo-epoxy-7 oxabicyclo[2.2.1]heptane derivatives. AB - 1 alpha,2 beta(5Z),3 beta(1E,3S),4 alpha,5 alpha,6 alpha]-7-[5,6-Epoxy-3- (3 cyclohexyl-3-hydroxy-3-methyl-1-propenyl)-7-oxabicyclo[2.2.1]-hept-2- yl]-5 heptenoic acid (31) and [1 alpha,2 beta(5Z),3 beta(1E,3S),4 alpha,5 alpha,6 alpha]-7-[5,6-epoxy-3-[3-hydroxy-5-(p-hydroxyphenyl)-1- pentenyl]-7 oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid (37) were found to be selective TxA2 antagonists at the platelet and pulmonary thromboxane receptors. An efficient stereospecific synthesis of these compounds and a series of structural analogues is described. Compounds 31 and 37 both inhibited the bronchoconstriction induced by arachidonic acid in the anesthetized guinea pig. PMID- 2966249 TI - Aggregated early intervention effects for Down's syndrome persons: patterning and longevity of benefits. AB - Pooled findings from 21 early intervention demonstration studies for Down's syndrome infants and children yield consistency of short-term benefits in the growth of finer motor skills, simple social repertoire and DQ/IQ scores, but conflicting evidence in support or not of benefits in the gross motor, linguistic and cognitive/academic domains. Support for the tenacity of gains, on follow-up to the early years of primary schooling, is disappointing. It is recommended that: (1) intervention programmers view the key working assumptions and ideological positions governing present practices more critically; (2) intervention curricula reflect the unique biological and behavioural properties of the syndrome, taking into account individual differences which are independent of etiological label; and (3) care delivery systems be based more fully on multidisciplinary collaboration, especially between the health sciences and education fields. PMID- 2966248 TI - Impaired monocyte macrophages function in patients with Crohn's disease. AB - To investigate the monocyte/macrophages function in patients with Crohn's disease (CD), phagocytosis, monocyte polykaryon formation and accessory cell function were studied. The number of yeast particles/monocyte in patients with CD was significantly higher (p less than 0.005) than that in healthy donors (HD). A significant increase (p less than 0.01) of fusion index was shown in CD patients in the absence of recombinant gamma-interferon (IFN gamma), while only a small amount of fusion was seen in HD. Accessory cell function of the monocytes from patients with CD was reduced, but 3H-thymidine incorporation was increased when the lymphocytes from patients with CD were cultured with the monocytes from HD. These results suggest that peripheral blood monocytes from patients with CD have abnormal functions, which may be involved in the pathogenesis of the granuloma and giant cell formation in CD patients. PMID- 2966250 TI - Mortality and survival of the Down's syndrome population in Western Australia. AB - From 1976 to 1984, there were 149 Down's syndrome live births in Western Australia. Where previously the coexistence of congenital heart disease was a major determinant of early mortality, current survival rates do not reflect this. Survival rates have significantly increased over the last decade: particularly so for those with coexistent congenital heart disease. Projected frequencies for Down's syndrome persons in Western Australia are expected to significantly increase. This rise in prevalence will have major economic effects on the planning and delivery of health care for this population. PMID- 2966252 TI - Assessment of cardiac size on portable chest films. AB - Using both anthropomorphic phantoms and human patients, the authors have shown that the effects on cardiothoracic (CT) ratio of changing from the posteroanterior (PA) to the anteroposterior (AP) projection are much greater than the effect of reducing the anode-to-film distance (AFD). This is because, in the AP projection, not only is the transverse diameter (TD) of the heart much farther from the film (and its image is therefore enlarged), but the TD of the chest is usually closer to the film and its image is reduced in size. These factors, operating in different directions, cause a major change in the CT ratio. The authors have shown that small hearts are magnified more than large hearts by changing from PA to AP, and that the position of the TD of the thorax (whether it is farther anterior or farther posterior) markedly affects what happens to the CT ratio when the AFD is changed. Because of this, it is impossible to predict accurately what effects a given AFD and projection will have on the CT ratio. However, using approximate correction factors (-12.5% of CT ratio for a 40 in AFD, AP film and -10% of CT ratio for a 72 in AFD, AP film) a clinically useful determination can be rapidly made as to whether the heart is enlarged or not. PMID- 2966251 TI - Hepatitis B in a hospital for the mentally subnormal in South Wales. AB - The resident population in a long-stay hospital for the mentally handicapped was surveyed in order to assess evidence for past and present infection with the Hepatitis B virus. The authors found a 0.5% prevalence of carriage of Hepatitis Be Antigen and do not recommend mass vaccination of staff and residents in this hospital. PMID- 2966254 TI - [Emergency 4-vessel aortocoronary bypass graft following acute coronary occlusion after pecutaneous transluminal coronary angioplasty: a case report]. PMID- 2966253 TI - [Percutaneous transluminal angioplasty of the subclavian and carotid arteries]. PMID- 2966255 TI - [Review of side effects of anti-tuberculosis drugs--with special reference to the endocrinological approach to acneiform eruptions on the skin]. PMID- 2966256 TI - First-use reactions during hemodialysis: a definition of subtypes. AB - We believe that it is worthwhile to separate out two distinct types of first-use reactions: a hypersensitivity type (type A) characterized by anaphylactic signs and symptoms and a non-specific type (type B) characterized primarily by chest pain and back pain. Further research is needed to better define the etiology, the epidemiology, and the pathogenesis of these two subgroups of the first-use syndrome. PMID- 2966257 TI - Dialyzer membranes: syndromes associated with first use and effects of multiple use. PMID- 2966258 TI - [Role of opioid peptides in the pathogenesis of vestibulo-autonomic disorders]. AB - The study was carried out using 12 noninbred male cats and 14 white rats. In response to vestibulo-autonomic disorders the rats showed a decrease of beta endorphin in the midbrain, medulla oblongata and hypothalamus as well as a reduction of met-enkephalin in the hypothalamus and medulla oblongata. The concentration of met-enkephalin in the adrenals increased and that of beta endorphin in blood did not change. This may be attributed to the intraneuronal redistribution of opioids and their transfer to the pituitary or release into the cerebrospinal fluid. Opioid variations give evidence that vestibuloautonomic disorders in rats do not stimulate the pituitary-adrenal system. The cats were exposed to vestibulo-autonomic disorders and subsequent intracerebroventricular administration of regulatory peptides or injection of opiate receptor blockers into the chemoreceptor trigger zone. It was demonstrated that naloxone, gamma endorphin and des-Tyr-gamma-endorphin were effective in protecting the vestibular function whereas ICI 154, 129 (a selective antagonist of delta-receptors) was practically ineffective. PMID- 2966259 TI - Suppressor cell generation during normal wound healing. AB - We have previously shown that 10 day healing wounds in rats contain wound mononuclear cells (WMNC) which inhibit normal lymphocyte mitogenic and allogeneic responses. In the present study we sought to further characterize the WMNC and define their mechanism of action. Polyvinyl alcohol sponges implanted in wounds were harvested and processed 10 days postwounding. The resultant WMNC suspension contained less than 15% macrophages. By FACS analysis, 69.5 +/- 11.4% (mean +/- SD of eight separate experiments) of the cells expressed the all T cell marker (W3/13), while 47.7 +/- 11.9% stained with the T helper/effector marker (W3/25) and 49.5 +/- 18.8% expressed the T suppressor/cytotoxic phenotype (OX8) (Th/Ts ratio = 0.96 +/- 0.13). When various numbers of WMNC were cocultured with 5 X 10(5) PHA-stimulated rat thymic lymphocytes, as few as 500 WMNC inhibit normal blastogenesis. Long-term (72 or 144 hr) culture of WMNC revealed that they maintain their suppressive activity. Furthermore, the conditioned media of long term cultures also significantly suppressed thymic lymphocyte PHA blastogenesis, suggesting that the WMNC secrete suppressive cytokines. Large doses of human recombinant IL-2 or indomethacin did not abrogate the inhibitory effect of WMNC. We conclude that the healing wound is normally infiltrated by suppressor lymphocytes which generate immune inhibitory cytokines. PMID- 2966260 TI - Quantitation of absolute flow in coronary artery bypass grafts using digital subtraction angiography. AB - Coronary angiography defines the location and size of obstructive lesions, but does not assess their physiological significance. To assess a new method to measure the blood-flow waveform, reversed saphenous vein grafts from the left subclavian artery to the left anterior descending coronary artery were placed in five mongrel dogs. Contrast material was injected selectively into the vein graft while obtaining fluoroscopic images from AP and 45 degrees LAO projections. Blood flow was measured under baseline, low-flow, and hyperemic conditions using an electromagnetic flow probe (EM). Seventeen radiographic determinations of mean blood flow (range 18-130 ml/min) were linearly correlated to simultaneous EM measurements (r = 0.91 and 0.88, respectively). Contrast material injections changed EM flow measurements by an average of 35%, which though large, is less than with other radiographic methods. The computed blood-flow waveforms had a time resolution of 1/30 sec and were in good agreement with EM waveforms measured simultaneously. Clinical application of this radiographic method for determining the blood-flow waveform may allow early prediction of coronary artery bypass graft closure. PMID- 2966261 TI - Blood flow velocity of internal mammary artery and saphenous vein grafts to the coronary arteries. AB - Doppler-derived blood flow velocity measurements were used to characterize the hemodynamics of 66 internal mammary artery grafts and 60 saphenous vein grafts to the coronary arteries at operation. Pulsed Doppler spectral analysis of centerstream graft flow demonstrated predominantly diastolic flow with a variable, multiphasic flow pattern in systole. The magnitude and configuration of the graft flow velocity waveform varied with graft type and whether the runoff was to single or multiple arteries. At operation, peak diastolic flow velocity was greater (P less than 0.0001) in internal mammary artery grafts to a single outflow artery (71 +/- 2 cm/sec) compared with single vein grafts (31 +/- 4 cm/sec). Sequential grafts demonstrated increased flow velocity and forward flow throughout the pulse cycle, indicative of low outflow resistance. Analysis of the phasic flow patterns permitted an assessment of functional graft patency. Technical errors (anastomotic stricture, internal mammary pedicle torsion) were identified in three grafts with low or absent diastolic flow. Vasospasm of the internal mammary artery was associated with high flow velocity throughout the pulse cycle. Observed differences in patency and the development of intimal hyperplasia between internal mammary artery and saphenous vein grafts may be related to graft hemodynamics. PMID- 2966262 TI - 17th annual UCLA symposium. Abstracts: Biological and molecular aspects of atrial factors. PMID- 2966263 TI - Coronary angioplasty: a cardiologist's perspective. PMID- 2966264 TI - Coronary bypass grafting after failed elective and failed emergent percutaneous angioplasty. Relative risks of emergent surgical intervention. AB - Emergency coronary artery bypass grafting after failed elective percutaneous transluminal coronary angioplasty can be performed with acceptable complication rates. Recently, however, a new class of patients with unsuccessful angioplasty has evolved with the use of thrombolytic therapy and emergent angioplasty as treatment for developing acute myocardial infarction. The efficacy of surgical intervention after failure of angioplasty in this setting has not been demonstrated. This report compares the results of coronary bypass done emergently after either failed elective or failed emergent angioplasty. Between March 1984 and September 1986; 1350 angioplasty procedures were performed at our institution, 393 for acute myocardial infarction. Of the 111 patients who came to operation, 42 had had unsuccessful elective angioplasty and 69 unsuccessful angioplasty done in the clinical setting of an evolving acute myocardial infarction detected by electrocardiographic criteria. Twenty-one of the 42 patients having unsuccessful elective angioplasty (group I) and 32 of the 69 with unsuccessful emergent angioplasty (group II) underwent emergency coronary artery bypass grafting. A retrospective nonparametric statistical comparison of the two groups was performed. Age, preoperative ejection fraction, distribution of vessels undergoing angioplasty, and number of vessels bypassed were not statistically different. All group II patients received thrombolytic therapy, and a reperfusion catheter was used in over half the patients in each group. Three group I and six group II patients required a preoperative balloon pump, and half the patients in each group required postoperative inotropic support. One patient in group I (4.7%) and two patients in group II (6.2%) died (no significant difference). Only five patients in group I (23.8%) and 11 in group II (34.3%) had enzymatic and electrocardiographic evidence of an acute myocardial infarction at discharge. Six patients in group II (15.6%) required reexploration for bleeding, versus none in group I (p = 0.04). Nonhemorrhagic complication rates, mean in patient and acute care days, total hospital charges, and blood product utilization rates were not statistically different. These data indicate that emergency coronary artery bypass grafting can be performed when necessary in the setting of failed emergent percutaneous transluminal coronary angioplasty with results comparable to coronary bypass after failed elective angioplasty. PMID- 2966266 TI - The interferon gamma receptor. AB - Interferon-gamma (IFN gamma) can regulate in vitro and in vivo a number of functions on a variety of cell types including some with no known immune functions. Recent work has indicated that IFN gamma exerts its effects upon interaction with a specific receptor on the cell surface. An IFN gamma receptor has been detected in a large variety of cell types of human and murine origins. Species specificity of IFN gamma appears to be a function of ligand-receptor interaction. In the human system, the IFN gamma receptor is a protein of 90 kd molecular weight. IFN gamma is internalized and degraded at a constant rate. The continuous uptake of IFN gamma is due to the presence of an intracellular receptor pool and to a mechanism of receptor recycling. By comparing the relationship between receptor occupancy and biologic response induction in macrophages, two activation mechanisms became apparent. Induction of certain functions such as H2O2 secretion, expression of Fc receptor or IA appeared to require only a single round of receptor occupancy. However, induction of more complex functions such as non-specific tumoricidal activity appeared to require three to four rounds of receptor occupancy. These results thus support the concept that IFN gamma induces different activities by triggering a different pathway at the cell surface and/or inside of the cell. PMID- 2966265 TI - Trends in invasive treatment of single-vessel and double-vessel coronary disease. AB - We examined our practice of invasive therapy for one- and two-vessel coronary disease to assess the impact of the randomized trials of coronary surgery and the current use of angioplasty. We first reviewed our results with coronary artery bypass graft in equivalent patients in the Coronary Artery Surgery Study with one and two-vessel disease between 1976 and 1981. Among 1376 patients, hospital mortality was 0.07%, and 5-year survival was 95.2% +/- 0.8%. To define trends in invasive therapy, which have since occurred, we compared 100 patients with one- and two-vessel disease in each of three groups: 1979 coronary artery bypass graft, 1984 coronary artery bypass graft, and 1984 percutaneous transluminal coronary angioplasty. Preoperative characteristics in the average 1979 and 1984 patients were similar; however, in 1984, patients who had a coronary artery bypass graft were older than patients who had percutaneous transluminal coronary angioplasty (61.5 versus 56.7 years, p less than 0.01), they required more heart medications (2.1 versus 1.5, p less than 0.01), had more previous infarctions (0.8 versus 0.5, p less than 0.01), and more patients had an ejection fraction of less than 50% (34% versus 7%, p less than 0.01). Patients who had angioplasty had a shorter postoperative stay (median number of days 7, 7, 2, p less than 0.01). Freedom from major complications was similar among the groups (91%, 87%, 85%). Unstable symptoms were the most frequent indication for invasive therapy (approximately 80%), whereas long-term symptoms, those considered in the randomized trials, occurred in relatively few patients. The number of patients without at least one definite indication for invasive therapy was 13%, 3%, and 11%, p less than 0.05, suggesting that the indications for the 1984 coronary artery bypass graft group have become more restrictive since the 1979 coronary artery bypass graft group. Indications for the 1984 percutaneous transluminal coronary angioplasty group remained less restrictive, being similar to those for the 1979 coronary artery bypass graft group. A continuing trend toward the use of percutaneous transluminal coronary angioplasty was evident, as 56% of the 1979 coronary artery bypass graft group of patients and 32% of the 1984 coronary bypass group of patients would be offered percutaneous transluminal coronary angioplasty rather than coronary artery bypass graft on the basis of 1986 percutaneous transluminal coronary angioplasty criteria. The p values were obtained with analysis of variance or chi 2 test. PMID- 2966267 TI - Heterogeneity of insulin and insulin-like growth factor I binding in a human Burkitt type ALL cell line during the cell cycle and in three Burkitt type ALL sublines. AB - The expression of hormonal and growth factor receptors in leukemic cell lines might be heterogeneous due to the admixture of different cell cycle phases and the presence of yet unidentified sublines. Therefore, cell cycle-specific separation of Burkitt type ALL cells was performed by counterflow elutriation, and by limited dilution procedure three sublines of this cell line were obtained. Counterflow elutriation enriched to 60-80% purity for G1-, S-, and G2-phase which was shown by DNA flow cytometry. Insulin and insulin-like growth factor I (IGF-I) binding was investigated in the G1, S, and G2 phase. Insulin binding sites decreased from 10 to 15,000 per cell in G1 to 1,000-5,000 in S and increased to 40-50,000 in G2. The affinity of insulin binding remained constant during the cell cycle. IGF-I binding sites increased from 2,000 per cell in G1 to 5,000 in S and 15,000 in G2. The affinity of IGF-I binding decreased from G1 toward S and then remained constant in G2. The three isolated clonal sublines differed in numbers of insulin and IGF-I binding sites/cell without differences in affinity. The fact that IGF-I shows higher affinity binding during G1 than during S and G2 Burkitt type ALL cells suggests that IGF-I might be important for initiation of proliferation. The reduction in insulin binding sites during S-phase may indicate refractoriness of the cell to insulin during DNA replication. PMID- 2966268 TI - The atrial natriuretic factor in hypertension. PMID- 2966269 TI - [Efficacy of laparoscopy in the diagnosis of ascites of indeterminate origin]. PMID- 2966270 TI - Beneficial actions of the thromboxane receptor antagonist, AH-23,848, in acute myocardial ischemia. AB - Thromboxane A2 (TxA2) production increases significantly during acute myocardial ischemia. Since TxA2 induces platelet aggregation, coronary vasoconstriction and has a direct cytolytic effect, thromboxane receptor antagonists would be expected to be beneficial in acute myocardial ischemia. A new thromboxane A2 receptor antagonist, AH-23,848, was studied in a cat model of acute myocardial ischemia. Myocardial ischemia was induced by ligation of the left anterior descending (LAD) coronary artery. Thirty minutes later, AH-23,848 or vehicle was given as a bolus (1 mg.kg-1) followed by a continuous infusion (1 mg.kg-1.h-1). AH-23,848 effectively reduced the S-T segment elevation while vehicle treated cats showed an increase. From direct myocardial biopsies, it was also seen that AH-23,848 prevented the loss of creatine kinase (CK) activity from the ischemic myocardium. Furthermore, the loss of amino-nitrogen compounds was also significantly reduced (p less than 0.05) by treatment with the receptor antagonist. This protective effect was not due to an indirect reduction of myocardial oxygen demand since blood pressure, heart rate or their product was unaltered by AH-23,848 administration. Moreover, the specificity of AH-23,848 to thromboxane receptors was confirmed in isolated cat coronary arteries and in cat platelets. These experiments demonstrate that blockade of the thromboxane receptor by AH-23,848 is an effective means of preventing acute myocardial ischemic damage in the cat, and thus thromboxane A2 plays a role in propagating the extension of ischemic damage during acute myocardial ischemia. PMID- 2966272 TI - [The "kissing" technic in coronary angioplasty]. PMID- 2966271 TI - Two monoclonal antibodies recognizing subpopulations of canine T lymphocytes with or without suppressor/cytotoxic functions. AB - Two monoclonal antibodies (mAb) were studied in regards to their ability to differentiate functionally distinct subsets of canine lymphocytes. MAb DT-2 (IgG2a) reacts with 33-55% of canine peripheral blood mononuclear cells (PBMC), which are also surface immunoglobulin (SIg) negative. Cytolytic treatment with DT 2 eliminates the ability of PBMC to respond to allogeneic cells and mitogens but does not affect alloantigen-primed cytotoxic or suppressor cells. MAb E11 (IgG3) reacts with 15-40% of PBMC which are also SIg negative. Cytolytic treatment with E11 affects responses of PBMC to allogeneic cells or mitogens to a lesser extent than DT-2 but eliminates cytotoxic and suppressor cells among alloantigen-primed lymphocytes. Thus, canine helper cells appear to be Thy 1+, 7.2+, DT-2+, DLy 6+, DLy 1+, E11-, SIg-, and cytotoxic/suppressor cells Thy 1+, 7.2+, E11+, DT-2+, DLy 6-, DLy 1-, SIg-. PMID- 2966273 TI - Concentration(s) of atrial natriuretic hormone in the plasma of rats with streptozotocin-induced diabetes mellitus. AB - Blood was withdrawn from rats injected 1, 3, 6 and 12 weeks previously with the diabetogenic agent streptozotocin (55 mg/kg, I.V.) or saline. Analysis of plasma showed that while the streptozotocin-treated animals displayed significantly (p less than 0.05) diminished triiodothyronine levels and significantly (p less than 0.05) elevated osmolalities at all time points after injection, immunoreactive ANF levels were unchanged. Thus, there would appear to be no direct relationship between plasma atrial natriuretic factor levels and plasma triiodothyronine levels or plasma osmolalities in the diabetic rat. PMID- 2966274 TI - Cardiac effects of fentanyl and sufentanil within clinical concentrations on the isolated heart. PMID- 2966276 TI - Dental evaluation and treatment prior to head and neck radiation. PMID- 2966277 TI - AIDS news that every dentist should know. PMID- 2966275 TI - To hospitalize or not to hospitalize? PMID- 2966278 TI - [Viremia in hedgehogs infected with the tick-borne encephalitis virus]. PMID- 2966279 TI - [Current status of foci of tick-borne relapsing fever in Kashka-Darya Province]. PMID- 2966281 TI - Ir gene control of help and suppression. PMID- 2966280 TI - [Surgical treatment and result of high aortic occlusion]. AB - Twenty-one patients with high aortic occlusion treated at our institution from 1967 to 1986 were reviewed. There were seventeen men aged from 39 to 78 (mean age: 61.0) underwent surgical intervention. All patients presented clinical manifestations of vascular insufficiency of the lower limbs; rest pain in eleven patients, intermittent claudication in nine and the others. Sexual impotence was present in eight patients. Renal artery involvement was seen in one case, and renovascular hypertension was observed in this patient. Hypertension and ischemic heart disease were present in twelve cases, cerebrovascular insufficiency in one case, diabetes mellitus in three cases. The following surgical treatments were performed; end-to-end Y-shaped Dacron graft implantations from the infrarenal abdominal aorta to the common femoral arteries in six patients, onlay V-shaped Dacron graft implantations in three patients, axillofemoral extra-anatomic bypass in four patients, and amputation only in one. The hospital mortality was 18% (3/17). Twelve patients discharged from the hospital are followed up (average period was 118 months), but the follow up was lost in two patients. There were two late deaths, which course was not related to operations. The prognosis of high aortic occlusion after anatomic bypass is good, thus it was concluded that anatomic bypass with Y-shaped or V-shaped Dacron graft was recommended and extra anatomic bypass might be performed only in a high risk patient. PMID- 2966283 TI - Selective induction of antigen nonspecific suppressor cells with normal gut epithelium as accessory cells. PMID- 2966282 TI - Oral tolerance and the induction of T cell unresponsiveness. PMID- 2966284 TI - Isotype regulation by helper T cells and lymphokines. PMID- 2966285 TI - The effect of ethanol upon early development in mice and rats. XII. The effect of chronic biparental or paternal alcohol intake upon preimplantation development in mice. PMID- 2966286 TI - The pathological process. I. A theoretical attempt at a definition of the pathologic concept. PMID- 2966287 TI - Ultrastructural aspects of bronchial epithelial dysplasias. PMID- 2966288 TI - DNA nuclear content in borderline ovarian tumors. PMID- 2966289 TI - The clinical importance of the calcaneal tendon vasculature (tendo calcaneus). PMID- 2966290 TI - Relationships between cancer site and personal habits based on an autopsy series. PMID- 2966291 TI - Morphological data regarding the efficiency of the omentoplasty to assure favourable conditions by prevention of the recurrent ureteral strictures in retroperitoneal fibrosis (an experimental study). PMID- 2966292 TI - Histoenzymological changes in the small intestine and liver of Wistar rats after intragastric administration of procaine-based drugs. PMID- 2966293 TI - Some considerations on the synaptic architectonics of the cerebral cortex. PMID- 2966294 TI - Damage-resistant DNA synthesis in eukaryotes. AB - The molecular basis of sensitivity of ionizing radiation and other damaging agents is not clearly defined in eukaryotes. While a large number of mutants have been described only a few have been demonstrated to have a defect in the repair of damage to DNA. An interesting characteristic of a sub-group of these mutants, in different species extending throughout the phylogenetic scale, is the presence of damage-resistant DNA synthesis. This phenomenon is observed in cells from individuals with the genetic disorder ataxia telangiectasia, in HeLa cells treated with fluorodeoxyuridine prior to UV irradiation, in mutants of the fungus Neurospora crassa, the slime mould Dictyostelium discoideum, the fruit fly Drosophila melanogaster and possibly in the "wasted" mouse mutant. In the case of ataxia telangiectasia sensitivity is only observed to ionizing radiation or radiomimetic chemicals whereas sensitivity to a wider spectrum of mutagens is reported for the lower eukaryotic mutants. In all cases a reduced inhibition of DNA synthesis is obtained after exposure to an agent to which the cell type is hypersensitive. It is unclear how damage-resistant DNA synthesis contributes to increased sensitivity in these cells, but is unlikely to be the major mechanism predisposing to radiation-induced cell death. The description of a derivative of an ataxia telangiectasia cell line with normal sensitivity to radiation but still maintaining resistant DNA synthesis partially uncouples radioresistant DNA synthesis and radiosensitivity. This paper is designed to review the phenomenon of damage-resistant DNA synthesis in a number of mutants. PMID- 2966295 TI - Fanconi's anemia lymphocytes: effect of caffeine, adenosine and niacinamide during G2 prophase. AB - In this investigation peripheral blood lymphocytes from 3 Fanconi's anemia (FA) patients, 2 FA heterozygotes and 4 normal subjects were treated with caffeine and/or adenosine, and/or niacinamide during G2 prophase. Caffeine dramatically increased breakage levels in homozygote and heterozygote cells. Niacinamide and adenosine decreased the amount of chromosomal aberrations detected in FA homozygote and heterozygote lymphocytes treated and untreated with caffeine during G2 prophase. Caffeine sensitivity of heterozygote lymphocytes is proposed as a new clinical test to explore heterozygosis in individuals of FA families. PMID- 2966296 TI - Effect of the homokaryotic state of the uvs-2 allele in Neurospora crassa on formaldehyde-induced killing and ad-3 mutation. AB - Formaldehyde was tested for its killing and mutagenic activities in the ad-3 forward-mutation test in Neurospora crassa. The test was conducted in 3 two component heterokaryons (dikaryons) of N. crassa in order to determine the effect of the uvs-2 allele, which causes a defect in nucleotide excision repair, on formaldehyde-induced killing and the induction of ad-3 mutants. These dikaryons were homokaryotic for uvs-2+ (H-12), homokaryotic for usv-2 (H-59), and heterokaryotic for uvs-2 (H-71). Formaldehyde induced killing and ad-3 mutants in H-12, but the presence of uvs-2 in the homokaryotic state (H-59) resulted in a 9 fold increase in killing and a 40-fold increase in the induction of ad-3 mutants. This increased sensitivity to formaldehyde-induced killing and mutation conferred by uvs-2 in the homokaryotic state (H-59 vs. H-12) is similar to that noted by others in Escherichia coli. Salmonella typhimurium and Saccharomyces cerevisiae. The dikaryon heterokaryotic for uvs-2 (H-71) has the same sensitivity to formaldehyde-induced ad-3 mutation as H-12, indicating that uvs-2 is recessive to uvs-2+. PMID- 2966297 TI - Inhibiting effects of nicotinamide on urethane-induced malformations and tumors in mice. AB - The antipellagratic vitamin, nicotinamide, significantly suppressed urethane induced malformations, when it was given intraperitoneally to pregnant JCL:ICR mice immediately after a single subcutaneous injection of urethane (1.0 mg/g) on the 9th day of gestation. The level of inhibition increased with the doses of nicotinamide: 33.0, 55.8, and 70.0% at doses of 0.1, 0.3, and 0.5 mg/g, respectively. Polydactyly and tail anomalies were markedly suppressed by the post treatment with nicotinamide, while cleft palates were less effectively suppressed. Nicotinamide was still effective, when it was given during the period of 24-48 h after urethane treatment. Furthermore, dietary administration of nicotinamide also reduced urethane-induced malformations. The level of inhibition was 39.4 and 61.1% at 0.5 and 1.0% of nicotinamide in the diet, respectively. Higher doses of nicotinamide (3 and 5% in diet) also inhibited urethane-induced malformations, but not so effectively as lower doses. The inhibiting effects of nicotinamide on the spontaneous incidence of cleft lips and palates in CL/Fr mice were significant at a low dose (0.5% in diet), but not at a higher dose (1.0%). When [carbonyl-14C]nicotinamide was given to pregnant mice, nicotinamide and small amounts of nicotinamide adenine dinucleotide (NAD+), but not nicotinic acid, were detected chromatographically in the fetus and placenta, indicating that nicotinamide or NAD+ acts directly on the fetus to suppress urethane-induced malformations. A preliminary study revealed that urethane-induced lung tumorigenesis in JCL:ICR mice was also inhibited by post-treatment with nicotinamide in the diet. The level of inhibition was proportional to the dose of nicotinamide, that is, 35.0 and 62.8% at 1.0 and 2.5% of nicotinamide in the diet, respectively. PMID- 2966298 TI - Effects of cell fusion and deoxynucleosides on sister-chromatid exchanges in B lymphoblastoid cell lines from 5 Bloom syndrome patients. AB - The effect of cell fusion and deoxynucleosides (deoxyadenosine, dA; deoxyguanosine, dG; deoxycytidine, dC; thymidine, T) on sister-chromatid exchanges (SCEs) in Bloom syndrome (BS) was studied in two types of BrdU (bromodeoxyuridine)-sensitive and BrdU-resistant B-lymphoblastoid cell lines (LCLs) with respect to cellular proliferation in BrdU-labeled culture conditions. Cell fusion between BrdU-sensitive and BrdU-resistant BS B-LCLs did not exhibit complementation, although when any of the BS B-LCLs (retaining high SCE character) labeled with BrdU were fused with non-labeled normal cells, the hybrid cells had a normal level of SCE at the first mitosis after fusion. Deoxycytidine addition showed no effect on SCEs in normal cells but decreased SCEs in BS cells from the baseline level of 70 SCEs/cell to about 60 SCE/cell. Purine deoxyribonucleosides (dG and dA) caused a significant concentration-dependent increase in SCE frequency both in normal and BS cells. Although T caused a 2-fold increase in normal SCEs, it highly decreased BS SCE from 70 SCEs/cell to 35 SCEs/cell. FrdU did not greatly affect BS SCE in the presence of BrdU and T. These observations indicate strongly that BS cells may have a low thymidine pool compared with normal cells, which could account for a more efficient BrdU substitution in the DNA thus potentiating the template effect on SCE. PMID- 2966299 TI - [Contact allergy for medications containing plant extracts]. PMID- 2966300 TI - [Percutaneous transluminal coronary angioplasty in patients with unstable angina pectoris]. PMID- 2966301 TI - [Effect of beta-endorphin on the development of denervation changes in the membrane of the muscle fiber in the rat]. AB - A decrease in the resting membrane potential, an increase in the input resistance, appearance of the anode breakdown and tetrodotoxin-resistant action potentials and extrajunctional acetylcholine sensitivity following denervation were revealed in surviving rat diaphragm muscle fibres. Addition of beta endorphin (1.10(-8) M) to the culture medium prevented on increase in the input resistance and reduced a degree of the extrajunctional acetylcholine sensitivity, but did not inhibit the appearance of the anode breakdown and tetrodotoxin resistant action potentials. Naloxone (1.25.10(-6) M) did not avert the effect of beta-endorphin, more over it exerted a beta-endorphin-like effect on the denervated muscle fibres. It is suggested that beta-endorphin can be the factor which is responsible for the neurotrophic control of passive electrical properties of the muscle fibre membrane and neurotrophic regulation of the membrane sensitivity to acetylcholine. PMID- 2966302 TI - Reconstructive surgery for renovascular hypertension secondary to arteriosclerosis and fibrodysplasia. III. The early and late effects of surgery on hypertensive target organ damage. PMID- 2966303 TI - Technical note: in vitro evaluation of an in-line Doppler flow-metering system for carotid artery shunts. AB - Carotid artery shunts are used extensively during carotid artery surgery to maintain cerebral perfusion. Blood flow through such shunts may be compromised by thrombosis, incorrect placement, or inadvertent clamping of the shunt. Currently, however, no direct method exists to detect poor shunt flow that might precipitate cerebral ischemia. A carotid artery shunt system that continuously monitors blood flow rates has been developed. This system utilizes a Doppler crystal embedded in the wall of a silicone elastomer shunt. The crystal ranges through a "liquid lens" that enables it to be placed without violation of the shunt lumen. Because the crystal is at a fixed angle (45 degrees) to the axis of blood flow and the diameter of the lumen remains constant, a linear relationship should exist between flow rates and the Doppler velocity signal. This hypothesis was tested in vitro using a pulsatile pump and both a starch-water solution and whole blood. Doppler velocity meter readings were compared to timed volume collections over a wide range of flow rates. A direct linear relationship between the Doppler flowmeter and timed volume collections existed, and the system was accurate to within 4.7%. This device may be useful in laboratory studies of carotid shunt dynamics and in clinical practice for early detection of correctable shunt flow abnormalities that could lead to cerebral injury. PMID- 2966304 TI - The effect of epileptiform discharges on neurohormonal release in epileptic patients with complex partial seizures. AB - We compared the nocturnal plasma prolactin (PRL) and beta-endorphin (B-E) concentrations prior to and after sleep deprivation (SD) in eight epileptic patients with complex partial seizures. After the period of SD (1) the mean number of interictal epileptiform discharges and the mean plasma PRL levels showed a significant rise during light non-REM stages of sleep, and (2) mean nocturnal plasma PRL and B-E concentrations showed a moderate rise during the first few hours of sleep, significant only for plasma PRL. In a patient with multiple complex partial seizures during sleep, the levels of plasma PRL and B-E concentrations were closely related to ictal discharges. The data obtained in this stress-free environment suggest a centrally mediated interaction between the release of PRL and B-E, in relation to epileptic discharges. PMID- 2966305 TI - Huntington's disease mortality in the United States. AB - The first analysis of multiple cause-of-death data for Huntington's disease in the United States was performed, using data from the National Center for Health Statistics for the period 1971 and 1973 through 1978. The overall mortality rate was 2.27 per million population per year, approximately 80% higher than the corresponding rate for deaths in which Huntington's disease was listed as the underlying cause of death. Age-specific mortality rates peaked around age 60. Rates were similar for both sexes, and higher in whites than nonwhites. Age adjusted rates were similar for United States whites and reported values from the Scandinavian countries. The leading causes of death were pneumonia and heart disease. Other common causes of death were nutritional deficiencies; mental disorders; cerebrovascular disorders; and accidents, poisonings, and violence. Suicide was rarely reported. PMID- 2966306 TI - Sarcoplasmic reticulum adenosine triphosphatase deficiency with probable autosomal dominant inheritance. AB - We report a family in which four members in two generations (mother, her son, and two daughters) suffered from impaired muscle relaxation aggravated by exercise. Muscle biopsies from two sisters showed moderate degree of histochemical type 2 fiber atrophy and excess of internal nuclei. Microscopic immunocytochemistry, using a monoclonal antibody raised against purified chicken SR-ATPase, revealed severe reduction of the immunoreactive ATPase of SR was markedly decreased on Western blots of muscle proteins. This family appears to have a clinically, electromyographically, and biochemically distinct metabolic myopathy associated with deficiency of SR-ATPase, with a probable autosomal dominant inheritance pattern that is phenotypically similar to recently described recessive cases. PMID- 2966308 TI - Protein gene product 9.5 expression in the lungs of humans and other mammals. Immunocytochemical detection in neuroepithelial bodies, neuroendocrine cells and nerves. AB - Immunoreactive protein gene product (PGP) 9.5 was demonstrated by immunocytochemistry in the innervation of human, monkey, pig, rabbit, rat, mouse and hamster lung. In most species PGP 9.5 immunoreactivity was furthermore obvious in the cytoplasm of the neuroepithelial bodies and neuroendocrine cells of the respiratory epithelium. Both a polyclonal and a monoclonal antibody to PGP 9.5 were applied and their characteristics compared. In several species, PGP 9.5 appears to be a useful histological marker for both the innervation and the diffuse neuroendocrine system (DNES)-type cells in the respiratory mucosa and might be added to the growing list of the pan-neuroendocrine markers. PMID- 2966307 TI - [Arterial hypertension with hypokalemia]. AB - After a careful review of the literature on physiopathological role of the renin angiotensin-aldosterone system in cases of arterial hypertension with hypokalaemia, a simple diagnostic procedure is suggested to differentiate between this and secondary hypertensions which, though few, are often curable. Three clinical cases examined in the medical department of Casalpusterlengo Hospital are presented in which the study of the renin-angiotensin system proved decisive for a correct diagnosis of the hypertension. PMID- 2966310 TI - Identification of a 5-HT1 recognition site in human brain membranes different from 5-HT1A, 5-HT1B and 5-HT1C sites. AB - In human caudate and cortex membranes, [3H]serotonin ([3H]5-HT) labels 5-HT1A and 5-HT1C recognition sites which show nanomolar affinity for 8-OH-DPAT (8-hydroxy-2 (di-n-propylamino)-tetralin) and mesulergine respectively, whereas no 5-HT1B binding could be identified. However, the majority of the sites labelled by [3H]5 HT (greater than or equal to 60% in cortex, 90% in caudate) are different from 5 HT1A, 5-HT1B and 5-HT1C sites. Competition experiments were performed in human caudate membranes incubated with [3H]5-HT in the presence of 100 nM 8-OH-DPAT and 100 nM mesulergine. Under those conditions, [3H]5-HT labelled an apparently homogeneous population of 5-HT1-like sites which display nanomolar affinity for tryptamines (5-carboxamido-tryptamine, (5-CT) greater than 5-HT greater than or equal to 5-methoxytryptamine (5-MeOT) greater than tryptamine) and some ergolines (metergoline greater than methysergide). In contrast, these sites showed low affinity for drugs with high affinity and/or selectivity for 5-HT1A (8-OH-DPAT, buspirone), 5-HT1B (21-009, RU 24969), 5-HT1C (mesulergine, mianserin) and 5-HT2 sites (ketanserin, cinanserin). The pharmacological profile of these sites is different from that of 5-HT1A, 5-HT1B, 5-HT1C, 5-HT2 and 5-HT3 sites but is consistent with the pharmacology of a 5-HT1-like receptor. It is very similar to that of the 5-HT1D site recently described in bovine brain by Heuring and Peroutka. PMID- 2966309 TI - D1- and D2-receptor antagonists induce catalepsy via different efferent striatal pathways [corrected]. AB - The D1- and D2-receptor antagonists SCH 23390 and raclopride produced a dose dependent catalepsy as studied by a vertical grid test in the male rat. The benzodiazepine antagonist Ro 15-1788 antagonized catalepsy induced by the D2 receptor antagonist raclopride while it failed to block the action of SCH 23390. The antimuscarinic drug scopolamine reduced and the gamma-aminobutyric acid (GABA) agonist muscimol enhanced catalepsy induced by both SCH 23390 and raclopride. The results suggest that D2-antagonist-induced catalepsy is mainly mediated via activation of the strio-pallidal GABA pathways whereas D1-receptor antagonist induced catalepsy is mainly mediated via increased activity in the nigrothalamic GABA pathways. PMID- 2966312 TI - Antibodies to calcium/phospholipid binding protein (calelectrin) recognize neurons, astrocytes and Schwann cells in the nervous system of rat. AB - We report on the identification of proteins Mr 32 kDa, 34 kDa, 68 kDa in rat brain or sciatic nerve which cross-react (by immunoblotting) with antibodies to purified Ca2+/phospholipid binding protein calelectrin from Torpedo marmorata. Immunocytochemical staining revealed that anti-calelectrin antibodies labeled cell bodies and processes of cortical neurons and astrocytes in the central nervous system (CNS). In sciatic nerve, calelectrin-like immunoreactivity was expressed in the cytoplasm of Schwann cells of adult as well as newborn rat. Additional staining occurred at periaxonal sites in mature nerve. The localization of calelectrin-related proteins in distinct cell types of the mammalian nervous system suggests a cell-specific role of this new group of proteins in Ca2+-mediated membrane processes involved in neural function. PMID- 2966311 TI - Intraparenchymal allografts in the mouse brain in relation to immunocytochemical identification of T lymphocyte subsets. AB - In a study of intracerebellar allografts of mice brainstem anlagen (embryonic day 12-14), we examined immunocytochemically the expression of two different types of T lymphocytes in and around the grafts. Helper/inducer and cytotoxic/suppressor T cells were identified with anti-L3T4 and anti-Lyt-2 monoclonal antibodies, respectively. Allografts into major histocompatibility complex (MHC)-compatible recipients showed no histological signs of rejection such as marked neovascularization and cellular infiltrates even 6 months after transplantation, but those into MHC-incompatible recipients generally had rejection reactions within one month after transplantation. In the latter cases, the L3T4/Lyt-2 ratio for the T lymphocytes in the infiltrates of the grafts was 1.03 +/- 0.14 (mean +/ S.D.), suggesting that both helper/inducer and cytotoxic/suppressor T cells may play important roles in the mediation of intraparenchymal brain allograft rejection. PMID- 2966313 TI - Serotonin-1A receptor activation in hippocampal CA1 neurons by 8-hydroxy-2-(di-n propylamino)tetralin, 5-methoxytryptamine and 5-hydroxytryptamine. AB - Serotonin (5-HT) usually induced a slow hyperpolarization lasting several minutes on first drop-application onto CA1 neurons. Subsequent applications always caused a briefer (less than 2 min) hyperpolarization, usually followed by a depolarization. 8-Hydroxy-2(di-n-propylamino)tetralin, a 5-HT1A receptor agonist, and 5-methoxytryptamine, a 5-HT1 receptor agonist, produced only the long-lasting hyperpolarization. The application of 5-HT agonists caused a persistent prolongation of the post-spike train afterhyperpolarization. These observations suggest that the long-lasting hyperpolarization produced by 5-HT may be mediated by the activation of the 5-HT1A receptor subtype. PMID- 2966314 TI - Tonic pain time-dependently affects beta-endorphin-like immunoreactivity in the ventral periaqueductal gray matter of the rat brain. AB - beta-Endorphin-like immunoreactivity (B-EP-LI) levels have been investigated in the ventral periaqueductal gray matter (vPAG) of rats killed 30, 60 or 120 min after the subcutaneous injection of dilute formalin (0.08 ml, 5%) in one fore- or hindpaw, or comparable handling. B-EP-LI was estimated by radioimmunoassay, using an anti-camel B-EP serum directed against the C-terminal portion of B-EP molecule. In both fore- and hindlimb groups vPAG B-EP-LI values were significantly increased 60 and 120 min after the injection relative to controls. Values from animals killed 120 min after formalin injection were higher than the ones at 30 and 60 min, forelimb effects being quantitatively more pronounced. The increase in B-EP-LI appeared distributed along the whole rostrocaudal extent of the region. PMID- 2966315 TI - Salvage PTCA in patients who are not candidates for coronary surgery. PMID- 2966316 TI - Chronic back pain. PMID- 2966317 TI - Dental care of AIDS patients with tuberculosis. PMID- 2966318 TI - [Information sources for hospital planning--perspectives of need-oriented indicator systems]. PMID- 2966319 TI - [Phenomenology and biographical method as a principle of social medicine assessment, especially of depressions]. PMID- 2966320 TI - [Changes in the pathogen spectrum in bacterial meningitis]. PMID- 2966321 TI - [AIDS and syphilis--parallel cases in history?]. PMID- 2966322 TI - [Organochlorine compounds in breast milk]. PMID- 2966323 TI - [Increased incidence of listerioses in Stuttgart 1984-1987]. PMID- 2966325 TI - [Incidence and significance of legionellas in air conditioning equipment]. PMID- 2966324 TI - [Recommendations for chemical disinfection of hospital rooms and spaces]. PMID- 2966326 TI - [What can a patient education brochure accomplish?]. PMID- 2966327 TI - [Treatments for children with behavioral disorders with therapeutic emphasis on educational experience]. PMID- 2966328 TI - Sequelae and dental management of the head and neck radiation patient. PMID- 2966329 TI - Dental evidence aids diagnosis of Prader-Willi syndrome. PMID- 2966330 TI - Dental implications of Alzheimer's disease. PMID- 2966331 TI - [Down's syndrome with unusual karyotype: iso-pseudo-dicentric chromosome 21]. PMID- 2966332 TI - Compensation and non-compensation chronic pain patients compared for DSM-III operational diagnoses. AB - Two hundred and eighty-three mixed chronic pain patients, consecutive admissions, were diagnostically evaluated as per DSM-III, Axis I, Axis II or personality type psychiatric operational criteria. Controlling for primary organic treatment diagnosis, age and race, statistical comparisons were made between male compensation patients (n = 93) and male non-compensation patients (n = 23) and between female compensation patients (n = 38) and female non-compensation patients (n = 28) for all DSM-III diagnoses. Male compensation patients were significantly overrepresented for these diagnostic groups: conversion disorder (somatosensory type); combined personality disorders; and passive-aggressive personality disorder. Male non-compensation patients were significantly overrepresented for these diagnostic groups: no diagnosis on Axis I; combined personality types; and compulsive personality type. Female compensation patients were significantly overrepresented for conversion disorder (somatosensory) only. Female non-compensation patients were significantly overrepresented for generalized anxiety disorder and combined anxiety syndromes. Compensation chronic pain patients may be at risk for some psychiatric disorders not previously identified: conversion disorder (somatosensory), and personality disorders. PMID- 2966333 TI - Neck and shoulder pain associated with hepatic arterial chemotherapy using an implantable infusion pump. PMID- 2966335 TI - [Flow cytometry in the detection of residual disease]. AB - Flow cytometry is a biological analysis technics whose applications may be used for minimal residual disease evaluation. From these applications we retained the DNA content measurement (with DNA index), surface markers analysis, and flow karyotyping. The threshold cell aneuploidy detection depend of the DNA index of leukemia cell population. The establishment of a relation between cell surface markers and malignant character of the cell is difficult to obtain. Study of CALLA antigen in leukemic patients have shown variations of antigenic density with different categories of studied patients. Malignant cells studied at the diagnosis or at the relapse of leukemia have a mean antigenic density higher than normal cells. This fact could be used to evaluate minimal residual disease as shown with recent clinical observations. PMID- 2966334 TI - Acute and chronic treatment with selective serotonin uptake inhibitors in mice: effects on nociceptive sensitivity and response to 5-methoxy-N,N dimethyltryptamine. AB - The tail-flick and increasing temperature hot-plate tests were employed to study the effects of acute or chronic treatment with zimelidine, alaproclate or chlorimipramine on nociception and response to 5-methoxy-N,N-dimethyltryptamine (5-MeODMT) in mice. A single dose of the serotonin (5-HT) uptake inhibitors produced antinociception in the hot-plate test but not in the tail-flick test. After chronic administration, reduced tail-flick latencies were demonstrated 24, 48, 72 and 144 h after withdrawal of zimelidine treatment, 48 h after withdrawal of alaproclate and 48 and 96 h after withdrawal of chlorimipramine treatment. The hot-plate response temperatures were slightly lowered after chronic zimelidine treatment but not after treatment with alaproclate or chlorimipramine. The response to 5-MeODMT was not altered by a single dose of the 5-HT uptake inhibitors, however, after withdrawal of chronic treatment this response was increased in the tail-flick test but not in the hot-plate test. It was concluded that acute and chronic treatment with 5-HT uptake inhibitors modulate nociception differently, and that chronic treatment induces supersensitivity of spinal postsynaptic 5-HT receptors. Different modulation of different 5-HT receptor subpopulations by these compounds may possibly contribute to the test-dependent results. PMID- 2966336 TI - [Changes in the ATPase activity of cell structures of the myocardium and brain in sudden cardiac death]. PMID- 2966337 TI - [Ambulatory care of children with diseases of the urinary system]. PMID- 2966338 TI - [Methods of increasing the effectiveness of the rehabilitation of obese children]. PMID- 2966340 TI - Brushing up on oral care. PMID- 2966341 TI - Carers: flexible services. PMID- 2966339 TI - In vitro splicing of adenovirus E1A transcripts: characterization of novel reactions and of multiple branch points abnormally far from the 3' splice site. AB - During the analysis of the in vitro alternative splicing of the natural E1A transcript of adenovirus, other minor reactions were detected (Schmitt et al., 1987, Cell 50, 31-39). We report here their characterization. The first reaction concerns the excision of a 216 nucleotide intron delineated by the 9S 5' splice site and a 3' splice site 216 nucleotides downstream. It can occur on the premRNA transcript and the 13S and 12S mRNA species. Strikingly, the reaction uses one of 3 branch points located 51, 55 or 59 residues upstream of the 3' splice site, a distance which is unusually long since all the branch points mapped up to now are located between 18-37 nucleotides of the 3' splice site. The dramatic accumulation of the corresponding lariat intermediates, likely related to this long spacing indicates that the second splicing step is relatively unefficient. The second kind of reaction analysed is a cryptic splicing which uses a 3' splice site generated by the junction of the 13S mRNA exons, and leads to the formation of psi 12S and psi 9S mRNAs. In vitro, this reaction occurs only from a 13S mRNA transcript, and not from the 13S mRNA newly formed in the splicing assay, consistent with what has been observed in vivo. Thus, both the well known alternative and the minor reactions occurring in vivo from E1A premRNA and mRNAs are detected in vitro, implying that most of the alternative splicing machinery is reconstituted in the in vitro system. PMID- 2966342 TI - Localization and identification of immunoreactive atrial natriuretic factor (ANF) in the frog ventricle. AB - The localization of ANF-like immunoreactivity in the ventricle of the frog Rana ridibunda was examined by the indirect immunofluorescence and the immunogold techniques, using an antiserum against synthetic ANF (Arg 101-Tyr 126). At the optic level, an appreciable number of positive cardiocytes was observed in the frog ventricle. Electron microscopic studies showed that all secretory granules present in ventricular cardiocytes contain immunoreactive ANF. The immunoreactive material has been characterized by Sephadex G-50 gel chromatography and reverse phase high performance liquid chromatography (RP-HPLC). After gel filtration, ANF like immunoreactivity eluted in 3 peaks. The major immunoreactive peak corresponded to high molecular weight material, while one peak co-eluted with synthetic ANF (Arg 101-Tyr 126). Further analysis of frog ventricular extracts by RP-HPLC revealed that the low molecular weight material has the same retention time as synthetic ANF, suggesting a high degree of sequence homology between amphibian and mammalian ANF. These results indicate that in amphibians, ventricular cardiocytes synthesize a peptide immunologically and chemically related to mammalian ANF. PMID- 2966343 TI - Identification of protease 3.4.24.11 as the major atrial natriuretic factor degrading enzyme in the rat kidney. AB - We have identified a metalloendoprotease from rat kidney cortex that cleaves the cysteine-phenylalanine bond (Cys7-Phe8) within the 17 amino acid ring structure of atrial natriuretic factor (ANF). Cleavage at this site represents the major ANF degradative activity in rat kidney, and is inhibited by the known metalloendoprotease inhibitors, thiorphan, phosphoramidon and zincov with IC50 values in the nanomolar range. Since these are specific inhibitors of protease 3.4.24.11, both protease 3.4.24.11 and ANF degrading activities were monitored during purification. Both activities copurified at each chromatographic step. Furthermore, purified protease 3.4.24.11 cleaved ANF specifically at the Cys7 Phe8 bond. It is concluded from this work that the major ANF degrading enzyme in rat kidney is protease 3.4.24.11. PMID- 2966344 TI - Atrial natriuretic peptide and vasopressin in human plasma. AB - Using a specific radioimmunoassay for atrial natriuretic peptide (ANP), plasma immunoreactive ANP was measured in 17 normal subjects and 83 patients with various diseases. Plasma ANP concentration in normal subjects was 14.1 +/- 1.7 pg/ml (mean +/- S.E.). Relatively high plasma ANP concentrations were detected in patients with diabetes mellitus, hyperthyroidism, atrial fibrillation and liver cirrhosis. Plasma ANP concentrations in the patients correlated positively with mean arterial blood pressure and plasma AVP concentrations. Plasma ANP concentrations in the patients also had positive correlations with left atrial dimension and left ventricular diastolic dimension determined by echocardiography. Another positive correlation was observed in the patients between plasma AVP concentrations and mean arterial blood pressure. These results suggest that ANP is a volume regulatory hormone but also that ANP may be involved in the blood pressure regulating system. PMID- 2966346 TI - Forms of immunoreactive beta-endorphin in the intermediate pituitary of the holostean fish, Amia calva. AB - Acid extracts of the intermediate pituitary of the holostean fish, Amia calva, were fractionated by gel filtration chromatography and analyzed with radioimmunoassays specific for N-acetylated beta-endorphin and C-terminally amidated alpha-MSH. In these extracts beta-endorphin-related immunoreactive material and alpha-MSH-related immunoreactive material were present in roughly equimolar amounts. The immunoreactive beta-endorphin-sized material was tested for opiate receptor binding activity using a beta-endorphin radioreceptor assay. The results of these studies were negative. The immunoreactive beta-endorphin sized material was further analyzed by cation exchange chromatography at pH 2.5. Two major and three minor peaks of immunoreactive material were isolated. Peak 5 exhibited a net charge of +7 at pH 2.5 and represented 53% of the total immunoreactivity recovered. Peak 2 with a net charge of +3 at this pH represented 38% of the total immunoreactivity recovered. The minor forms, Peaks 1, 3 and 4, exhibited net charges of +2, +4 and +6, respectively. The apparent molecular weights of Peaks 2 and 5 were determined on a Sephadex G-50 column. Peak 2 had an apparent molecular weight of 2.7 Kd and Peak 5 had an apparent molecular weight of 3.5 Kd. Reverse phase HPLC analysis of Peak 5 indicates that this form of Amia beta-endorphin had chromatographic properties similar to salmon beta-endorphin II. These results would suggest that N-terminal acetylation and C-terminal proteolytic cleavage are important post-translational modifications of the forms of Amia beta-endorphin. PMID- 2966347 TI - [The role of magnesium in calcium lithiasis]. PMID- 2966345 TI - NH2-terminal fragment of rat pro-atrial natriuretic factor in the circulation: identification, radioimmunoassay and half-life. AB - A radioimmunoassay was developed to measure the NH2-terminal counterpart of rat pro-atrial natriuretic factor (pro-ANF) in plasma. Synthetic rat ANF (Asp 11-Ala 37) coupled to bovine serum albumin was used to immunize New Zealand rabbits. The antiserum demonstrated good immunoreactivity towards rat ANF (Asn 1-Arg 98), (Asn 1-Tyr 126), (Asp 11-Ala 37) and even human ANF (Asn 1-Ser 30). The standard curve had an ED80 of 9.5 +/- 2.5 and ED50 of 44.0 +/- 10.5 fmol/tube. Immunoreactive ANF NH2-terminal peptide was measured directly in rat plasma without prior extraction. In fact, extraction of ANF NH2-terminal from plasma by C18 silica gel chromatography revealed inconsistent recovery and a lack of parallelism. Morphine (0.75 mg/100 g), chosen to elicit increased ANF (Ser 99-Tyr 126) secretion, elevated its plasma concentration from 54.1 +/- 3.2 to 190.8 +/- 55.8 fmol/ml after 20 min. At the same time, the immunoreactive NH2-terminal fragment rose from 378 +/- 16 to 1181 +/- 201 fmol/ml. The identity of this immunoreactive material was verified following affinity chromatography and reverse-phase high performance liquid chromatography (HPLC) of plasma from morphine-treated rats. Molecular sieving and amino acid sequencing demonstrated that it appears to be consistent with or identical to rat ANF (Asn 1-Arg 98). The disappearance rate of ANF (Asn 1-Arg 98) was studied by injecting radioactive material into anesthetized rats. The exponential decay was analyzed by a two-compartment model in which the fast and slow components had a half-life of 2.5 +/- 0.3 and 54.8 +/- 3.9 min, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966348 TI - [Immune disorders in liver disease of alcoholic origin]. PMID- 2966349 TI - [Percutaneous placement of ureteral prostheses. 150 cases]. AB - Percutaneous placement of ureteral stents was attempted on 150 occasions with an 85 p. 100 success rate. Whether released or not, this type of prosthesis ensures that the urine is drained internally from kidney to bladder. Stenting is effective as palliative treatment of malignant stenosis or curative treatment of benign stenosis, as it maintains the ureteral lumen diameter after balloon catheter dilatation. It also dries up post-operative urinary fistulae and it controls and promotes their healing. Because of the satisfactory results obtained, the percutaneous technique is increasingly preferred to surgery performed under difficult conditions. PMID- 2966350 TI - [Acute cholecystitis after placement of biliary endoprosthesis. Treatment by percutaneous cholecystostomy]. AB - Seven cases of acute cholecystitis (4 women and 3 men, mean age 73 years) were observed in a series of 192 patients treated by endoprosthesis for cancerous obstruction of the bile ducts between october, 1984 and october, 1986. The suspected clinical diagnosis was confirmed by ultrasonography. Cholecystostomy was performed by percutaneous puncture under ultrasonic guidance. A catheter was positioned in the gallbladder by the Seldinger technique in 3 cases and by the trocar technique in 4 cases. Pain in the right hypochondrium was relieved in all patients immediately after emptying of the gallbladder. Five patients were cured. One patient developed purulent peritonitis which was treated by surgery. A female patient died of her pancreatic cancer 3 days after cholecystostomy. Provided a number of precautions are taken to prevent leakage of the infected bile into the peritoneal cavity, percutaneous cholecystostomy is the treatment of choice for acute cholecystitis consecutive to biliary endoprosthesis. PMID- 2966351 TI - [Evaluation of the methodological quality of randomized therapeutic trials]. AB - A grid devised to evaluate the methodological quality of randomized therapeutic trials is presented. Fourteen items are analysed: description of the principal criterion of assessment, criteria of inclusion, number of subjects seen and excluded, number of subjects randomized to each group, number of subjects excluded who are alive or lost to follow-up and reasons for the fall-outs, blind character of the doctors, patients and persons responsible for the assessment criteria, calculation of the number of subjects required before starting the trial, method of randomization and its blind character, analysis and discussion of covariables, statistical tests used, taking into account of the fall-outs and power of the trials with non-significant results. This grid, used for the analysis of 527 randomized trials, showed that about one-half of them were of mediocre quality, the most frequent defects encountered being the multiplicity of assessment criteria, the lack of description of the subjects excluded, the absence of calculation of the number of subjects required before starting the trial and the small number of subjects in trials with non-significant results. PMID- 2966352 TI - [Clinical value of the determination of urinary antitrypsin activity]. AB - Urinary trypsin inhibitory capacity is mainly due to the excretion of a glycoprotein which is immunologically related to the inter alpha-trypsin inhibitor and may be a proteolytic degradation product of that substance. It was tested in 133 subjects divided into 7 groups: 24 healthy controls (group A), 21 patients with bacterial infection (group B), 37 with bacterial infection under antibiotic therapy (group C), 25 with connective tissue disease (group D), 8 with infected connective tissue disease (group E), 14 with cancer (group F) and 4 with infected cancer (group G). Urinary trypsin inhibitory capacity level was very low in controls (3.32 +/- 0.8 U/g urinary creatinine), but it was dramatically increased when infection was present (149.67 +/- 23.6 U/g urinary creatinine). This test appeared to be more effective than serum C-protein measurement simultaneous carried out in the same patients. Urinary trypsin inhibitory capacity is not related to the degree of proteinuria in the urine sample, but it is increased in patients with chronic renal failure excluded from this study. Thus, its measurement is a sensitive, easy and useful test for detecting and monitoring infections. The return to its physiological value is a very good argument in favour of therapeutic effectiveness. PMID- 2966353 TI - [Left ventricular hypertrophy in arterial hypertension]. AB - Arterial hypertension is frequently and at an early stage complicated by left ventricular hypertrophy, i.e. an increase in muscular mass due to the proliferation of myofibrillae. This in fact is a physiological mechanism aimed at maintaining systolic function and systemic blood flow rate. Left ventricular hypertrophy may be associated with myocardial alterations, such as increase of collagen, abnormalities of diastolic function, reduced contractility, increased cell excitability and disorders of coronary perfusion. It is responsible for a higher risk of cardiovascular mortality. Antihypertensive treatments, therefore, must not only bring blood pressure down to normal values, but also reduce the myocardial mass. In order to avoid a detrimental effect on coronary reserve, it is highly desirable that arterial hypertension and left ventricular hypertrophy regress simultaneously. Regression of the myocardial hypertrophy associated with arterial hypertension is observed with most antihypertensive drugs, except vasodilators that act directly on the vascular smooth muscle, probably due to stimulation of the sympathetic system. Diuretics also have an inconstant beneficial effect on left ventricular hypertrophy. When a choice has to be made between two drugs that have the same antihypertensive activity, it is the one that also brings about an early and lasting regression of myocardial hypertrophy which must be prescribed. PMID- 2966354 TI - [Actinomycosis on an intra-uterine device: what is the cause?]. PMID- 2966355 TI - [Value of theophylline in the prevention of postoperative hypoxemia in abdominal surgery]. PMID- 2966356 TI - [Acromegaly with subnormal growth hormone in a diabetic]. PMID- 2966357 TI - [Comparative bacteremia in septicemic children with catheters in pediatric resuscitation]. PMID- 2966358 TI - [A totally implantable venous catheter in infection. Preliminary results]. PMID- 2966359 TI - [High-dose ketoprofen in surgery of the spine]. PMID- 2966360 TI - [Lyme disease with multiple joint involvement and a major inflammatory syndrome]. PMID- 2966361 TI - [Reconstruction of the posterior wall of the pharynx. Free transplants with vascular microanastomoses]. PMID- 2966362 TI - [Hygromas of the elbow and HIV infection. 6 cases]. PMID- 2966363 TI - [Macular anetoderma and primary Gougerot-Sjogren syndrome]. PMID- 2966364 TI - [Insulin of man and human insulins. Medical and economic aspects in 1987]. PMID- 2966365 TI - [Metabolism of proteins and postnatal growth in the premature infant]. PMID- 2966366 TI - [Peripheral bronchopulmonary cancers. Diagnostic efficiency of fibroscopic samples. Prospective study on 561 patients]. AB - The purpose of this french multicentric study was to evaluate the diagnostic accuracy of samples collected by fibroscopy in peripheral lung cancer with normal endoscopy. Five hundred and sixty-one patients entered the study; a tumour had been diagnosed in 350 of them (62 p. 100). Among these 350 patients, 147 were examined with the help of a light-amplifier screen, and a positive diagnosis was made in 97 (66 p. 100). Guided sample collection in the pathological area (biopsy, brushing, transcatheter aspiration) proved much more rewarding than unguided sample collection (biopsy of the bronchus, simple aspiration or cytology of 3-day sputum): 45 p. 100 positive results versus 18 p. 100 (P less than 0.001). However, sputum cytology still had good diagnostic value since it provided by itself the diagnosis in 14 p. 100 of the cases. Results were significantly inferior in tumours less than 3 cm in diameter. PMID- 2966367 TI - [Vesico-ureteral-renal reflux in the adult. Preliminary results of endoscopic treatment]. AB - Vesico-ureteric reflux in adult patient may now be treated endoscopically by injecting teflon under the ureteral orifices. Twenty-six ureteral units were treated by this technique in 18 patients. Correction of the reflux was obtained in 22 units (85 p. 100); the technique was ineffective in 4 units (15 p. 100). No complication was observed in this series; in particular, there was no stenosis of the intramural ureter. These encouraging preliminary results have to be confirmed on a long-term basis in large series of patients. PMID- 2966368 TI - [A simple test for the detection of respiratory allergy to current pneumallergens]. AB - Phadiatop is a new in vitro test used for screening patients with allergic respiratory diseases. This test, based on the RAST procedure, is performed with a paper disc to which a balanced mixture of relevant inhalant allergens has been coupled. One hundred and one adults ranging in age from 18 years to 73 years and 38 children (4 months to 15 years) were studied. We found a good correlation between this new test and total IgE (adults: 74.3 p. 100, children: 68.4 p. 100), specific IgE (adults: 91.1 p. 100, children: 100 p. 100) and skin test (adults: 79.2 p. 100, children: 97.1 p. 100). There was a high correlation between clinical atopy assessment and the result of Phadiatop, its efficiency was 83 p. 100 in adults and 97 p. 100 in children. PMID- 2966369 TI - [The implantable defibrillator. Long-term clinical results]. AB - Between october, 1982 and december, 1986, 17 patients were implanted with an automatic defibrillator. The thoracic route was used in 6 patients with installation of an epicardial patch and an endocavitary electrode. The subcostal route was used in the remaining 11 patients, with 2 epicardial patches. The device, badly tolerated by 2 of the first 6 patients, created no problem in the 11 patients of the second group. One patient died of pulmonary embolism on the 6th post-operative day. At follow-up, there was no inappropriate triggering of the AID-B: 8 patients had neither arrhythmia nor shock, and 8 received from 2 to 55 shocks with documented arrhythmia. Two patients died suddenly: one after end of life of the device, the other after 3 years, while the device was still working. The implantable defibrillator technique has improved and local complications are now seldom observed. The device is reliable and constitutes an effective weapon against sudden death caused by ventricular arrhythmia. PMID- 2966370 TI - [Thyroid ectopia and Basedow's disease]. AB - Graves' disease was diagnosed in a 35-year old woman with a history of myxoedema in childhood. Clinical data, ultrasonography and radioisotope scanning showed that the disease had developed in an ectopic subhyoid thyroid gland. Although numerous thyroid diseases associated with an ectopic thyroid have been described, Graves' disease does not seem to be frequent since only one case has been published so far. PMID- 2966371 TI - [Management in cases of accidental contamination by iodine radioisotopes]. AB - Iodide can reduce radioactive iodine thyroid uptake and whole-body irradiation. Maximal effectiveness is obtained when 3 conditions are fulfilled: adequate dosage: potassium iodide 130 mg (i.e. iodide 100 mg) as tablets in adults and older children, 50 mg in infants under one year of age; prompt administration after contamination and daily treatment as long as the contamination persists. Depending on the importance and duration of the contamination, different dosages may be required. In any case, children must be given priority for prophylactic measures. PMID- 2966372 TI - [A rare but severe accident in ascending phlebography: skin necrosis]. PMID- 2966373 TI - [Pneumopathies caused by metapramine]. PMID- 2966374 TI - [First French controlled trial on nicotine-releasing chewing gum versus placebo gum]. PMID- 2966376 TI - [Myasthenia appearing after the ablation of a thymoma. Two cases]. PMID- 2966375 TI - [Intra-operative anaphylactic shock caused by the latex in surgical gloves. 5 cases]. PMID- 2966377 TI - [Multifactorial etiology of overdose in antivitamin K administration]. PMID- 2966378 TI - [How does one develop an autoimmune disease? Probable pathogenic role of a membrane protein associated with systemic lupus erythematosus]. PMID- 2966379 TI - [Pharmacokinetics of antibiotics in the premature infant]. PMID- 2966380 TI - [A rare cause of hemoperitoneum. Spontaneous rupture of a hepatic artery, the site of septic thrombosis]. AB - An exceptional case of spontaneous haemoperitoneum due to rupture of a thrombotic hepatic artery is reported. The thrombosis was of infectious origin and due to an untreated 3-week old dental abscess. The different causes of such thromboses, the different steps in the diagnostic approach, the consequences of hepatic artery obstruction and the different therapeutic possibilities are discussed. PMID- 2966381 TI - [Incidence of waterborne lead poisoning in an internal medicine department in a region with acid soil]. AB - Between 1980 and 1985 patients living in the Vosges mountains and admitted to the internal medicine department of a university hospital were systematically investigated for chronic lead poisoning. The delta-aminolevulinic acid dehydrase assay was used for screening, and when necessary confirmation was obtained with calcium EDTA-induced plumburia test and lead assays in domestic water. Among 366 patients with no suspicion of lead poisoning, the condition was proven in 52 cases and found probable in 15 cases, i.e. in 18 p. 100 of these patients. Obviously, figures cannot be extrapolated to the entire population of the Vosges, but they indicate that lead-induced morbidity is widespread and that saturnism constitutes a major health problem. In one-half of the cases, chronic lead poisoning affected people over 65 while patients of that age group accounted for only 14 p. 100 of non lead-poisoned patients. Prolonged tolerance to the intoxication explains why saturnism has for so long passed unnoticed in the Vosges region, and it may be feared that the same applies to other regions with the same geological features as the Vosges. PMID- 2966382 TI - [Influence of pregnancy on the development of Hodgkin's disease]. AB - This retrospective study concerns 13 patients in whom Hodgkin's disease was diagnosed during pregnancy or immediately after delivery (group I) and 12 patients with Hodgkin's disease who had one or several pregnancies while under treatment (group II). In group I, Hodgkin's disease was diagnosed in early pregnancy in 4 patients who all had therapeutic abortion: 3 remain in prolonged complete remission and 1 had a late relapse; 9 cases were diagnosed in late pregnancy or after delivery: 3 were treatment failures, 2 had a relapse and 4 remain in complete remission. In group II patients, 3 pregnancies occurred during initial chemotherapy and were interrupted; 5 pregnancies occurred during subsequent radiotherapy or (for earlier patients) maintenance chemotherapy, and 4 of them were interrupted; 9 pregnancies occurred within 2 years of completing treatment, and 7 after 2 years. Of the 12 patients in group II, only 2 had a relapse whereas 10 remain in complete remission. Although they should be interpreted with caution, these data suggest that Hodgkin's disease diagnosed in late pregnancy or after delivery might be more active, and they justify therapeutic abortion when diagnosis is made in early pregnancy. They do not indicate a high risk of relapse in treated Hodgkin's disease patients during a subsequent pregnancy, even if it occurs shortly after treatment. PMID- 2966383 TI - [Cerebral protection in carotid artery surgery. The role of the intraluminal shunt. Results of a national survey]. AB - Clamping of the carotid artery made necessary by surgery of that vessel produces haemodynamic disturbances which may result in a neurological deficit accident at recovery from anaesthesia. Evaluating the patient's tolerance to carotid clamping and using brain protection methods during clamping increase the safety of the operation. The different methods of protection used in France were examined in an enquiry conducted among members of the French college of vascular surgery, with special attention to measurement of residual pressure in the clamped internal carotid artery (performed by 60 p. 100 of surgeons) and its corollary which is intraluminal shunting in case of hypotension. This type of enquiry concerning controverted technical procedures should provide information on the best way of improving surgical results. PMID- 2966384 TI - [Retroperitoneal fibrosis]. AB - Neoplastic and non-neoplastic retroperitoneal fibrosis share the same macroscopic and histological features and the same clinical signs which mainly consist of abdominal pain followed by signs of compression. All carry a risk of extension to adjacent structures and may therefore produce serious complications. They may also be associated with a "systemic" syndrome with fever, altered general condition and high erythrocyte sedimentation rate. The necessary laboratory and radiological examinations are detailed. Non-neoplastic retroperitoneal fibrosis may be due to a variety of causes, the most frequent of which today are medications and aortic atheroma. Surgery is usually needed when the disease is life-threatening or of poor functional prognosis. Some authors have suggested that surgery should be combined with corticosteroid therapy. In some cases, regression of the fibrosis could only be obtained by giving steroids in high doses. PMID- 2966385 TI - [A case of porocephalosis disclosed by echography during evaluation of hepatic amebiasis]. PMID- 2966386 TI - [Preparation for surgery of a pheochromocytoma with a calcium channel blockader]. PMID- 2966387 TI - [Detection and surveillance of women at risk of HIV infection attending a maternity clinic in Paris. A 24-month evaluation]. PMID- 2966388 TI - [Convulsions 12 hours after the ingestion of benzodiazepine with a short half life]. PMID- 2966389 TI - [Sequential high-dose corticosteroid therapy in multiple myeloma]. PMID- 2966390 TI - [Optic neuropathy in Mediterranean boutonneuse fever]. PMID- 2966391 TI - [Pulmonary sarcoidosis with bullous development]. PMID- 2966393 TI - [Reasons for the repeat hospital admission of patients with active pulmonary tuberculosis and the characteristics of their treatment]. PMID- 2966394 TI - [Complicated forms of tuberculosis in children and adolescents today]. PMID- 2966392 TI - ["Antisorbitol" preparations: problems and prospects (review of the literature and personal data)]. PMID- 2966395 TI - [Reasons for poor-grade fluorograms]. PMID- 2966396 TI - [Mass fluorography in polyclinics]. PMID- 2966397 TI - A C-terminal, calmodulin-like regulatory domain from the plasma membrane Ca2+ pumping ATPase. AB - A cDNA that encodes what appears to be the inhibitory domain of the plasma membrane calcium-pumping ATPase (Ca2+-ATPase) has been isolated by screening a lambda gt11 bovine brain cDNA library with antibodies prepared against the human erythrocyte membrane Ca2+-ATPase. This screening resulted in isolation of a bacteriophage containing a 1.5-kilobase cDNA insert encoding a 71-residue polypeptide, the remainder being a large 3' terminal noncoding region. A portion of this deduced peptide sequence was identical to that of a peptide isolated from a V8 protease digest of the human erythrocyte Ca2+-ATPase except for 1 residue. Antibodies purified by immunoabsorption to the fusion protein containing this cDNA-encoded polypeptide reacted only with those fragments of a limited trypsin digest of the human erythrocyte Ca2+-ATPase that contain the inhibitory domain. Moreover, these antibodies were able to partially stimulate basal enzyme activity and block further activation by calmodulin. The encoded polypeptide bears homology to the glutamic acid-rich regions N-terminal to the Ca2+-binding loops of calmodulin and to a lesser extent with the loops themselves. This encoded polypeptide also represents the C terminus of the Ca2+-ATPase. Portions of the isolated cDNA were homologous to the 3' noncoding region of the sarcoplasmic reticulum Ca2+-ATPase cDNA, indicating a possible mechanism for the evolution of these distinct membrane Ca2+ pumps. PMID- 2966398 TI - Calcium influx and the Ca2+-calmodulin complex are involved in interferon-gamma induced expression of HLA class II molecules on HL-60 cells. AB - Interferon gamma (IFN-gamma) induces HLA-DR and -DQ molecules and causes an accumulation of transcripts in HL-60 cells. Experiments were, therefore, designed to investigate the intracellular signaling molecules regulating the appearance of HLA class II molecules. The expression of HLA class II (DR and DQ) molecules induced by IFN-gamma was blocked by a calmodulin antagonist, W7, but not by a protein kinase C inhibitor, H7. Furthermore, a direct activator of protein kinase C, phorbol 12-myristate 13-acetate, was unable to induce HLA class II (DR) molecule expression. These results suggest that IFN-gamma induces HLA class II molecules on HL-60 cells by way of a calcium-calmodulin pathway and not by way of a protein kinase C pathway. Calmodulin is activated by a transient rise in the cytosolic free calcium. In fact, IFN-gamma evoked a calcium influx into HL-60 cells, whereas depletion of Ca2+ from culture medium resulted in a failure of IFN gamma to induce DR expression. Furthermore, the calcium ionophore A23187 by itself induced DR molecule expression. These results suggest that IFN-gamma stimulates calcium influx by a so-called receptor-mediated calcium channel and activates the calmodulin branch of the calcium messenger system, resulting in the induction of DR molecules on the surface of HL-60 cells. PMID- 2966399 TI - Conversion of encapsulated 5-fluoro-2'-deoxyuridine 5'-monophosphate to the antineoplastic drug 5-fluoro-2'-deoxyuridine in human erythrocytes. AB - The fluoropyrimidine deoxyribonucleotide 5-fluoro-2'-deoxyuridine 5' monophosphate (FdUMP) was encapsulated in human erythrocytes by a procedure based on hypotonic hemolysis and isotonic resealing. Encapsulated FdUMP (up to 9 mumol/ml of packed erythrocytes) did not affect erythrocyte metabolism or morphology. Hemolysates were found to catalyze efficient dephosphorylation of FdUMP to yield nearly stoichiometric amounts of the corresponding deoxyribonucleoside 5-fluoro-2'-deoxyuridine (FdUrd), an antineoplastic drug showing selective cytotoxicity toward liver metastases from colorectal carcinomas. The dephosphorylation reaction had an apparent Km of 7.7 +/- 1.2 mM FdUMP at pH 7.4 and was remarkably slower at pH 8.2. ATP, GTP, and UTP inhibited both the disappearance of FdUMP and the formation of FdUrd in hemolysates. The enzyme responsible for the FdUMP-to-FdUrd conversion was identified with the deoxyribonucleotide-specific isozyme of erythrocyte pyrimidine 5'-nucleotidase (EC 3.1.3.5). Intracellular formation and subsequent release of FdUrd were observed in intact erythrocytes loaded with FdUMP. Inhibition of FdUrd release from these erythrocytes was obtained by raising the pH intracellularly and, alternatively, by coencapsulation of ATP. Autologous FdUMP-loaded erythrocytes might be used as endogenous bioreactors designed for time-programmed and liver targeted delivery of FdUrd. PMID- 2966401 TI - Effect of Ca2+ on cross-bridge turnover kinetics in skinned single rabbit psoas fibers: implications for regulation of muscle contraction. AB - The effect of Ca2+ upon the rate constant of force redevelopment following a period of isotonic shortening with immediate restretch to the starting sarcomere length was studied in rabbit psoas fibers at 5 degrees C. Control experiments support the assumption that the rate constant of force redevelopment represents isometric cross-bridge turnover kinetics (fapp + gapp), where fapp and gapp are the rate constants characterizing the transitions from the non-force-generating states to the force-generating states and back to the non-force-generating states, respectively. Parallel measurements of the rate constant of force redevelopment and of force, stiffness, and fiber ATPase during isometric contraction allow the effect of Ca2+ upon fapp and gapp to be determined. Analysis reveals that Ca2+ has a marked effect upon fapp, while gapp remains approximately unchanged. Furthermore, in the range above 25-30% of maximum Ca2+ activation, regulation of force, stiffness, and ATPase is mediated through changes in fapp. Below this range, however, it cannot be ruled out that, in addition, cross-bridges are also switched in and out of the turnover process ("recruitment"). As a consequence of regulation through turnover kinetics, both Ca2+ sensitivity and the slope of force-pCa (-log[Ca2+]) relations are shown to be affected by the ratio fapp/gapp, which may represent an important mechanism of modulation of contractile function in addition to modulation through changes within the regulatory protein system. PMID- 2966400 TI - Paradoxical relationship between atriopeptin plasma levels and diuresis natriuresis induced by acute volume expansion. AB - Surgical removal of one or both atrial appendages was employed in rats to reduce the intrinsic stores of atriopeptin (AP). In conscious rats (with intact baroreceptor reflexes), bilateral or unilateral atrial appendectomy suppressed the diuresis and natriuresis produced by acute volume expansion. Surprisingly, volume expansion (with 4% bovine serum albumin in saline at 1.5 ml/kg per min for 15 min) did not result in an increase in plasma AP immunoreactivity (APir) in control or atrial-appendectomized conscious rats. Previous studies demonstrated that acute volume expansion in anesthetized animals caused increased plasma APir. Indeed, we found that volume expansion causes comparable diuresis-natriuresis in conscious and chloral hydrate-anesthetized rats, but only the latter group exhibits an increase in plasma APir. Brattleboro rats, which are deficient in vasopressin, exhibit the same response as Long-Evans controls in that acute volume expansion in conscious animals produces a pronounced diuresis and natriuresis but no APir release, but when these same animals are anesthetized, there is a simultaneous induction of diuresis-natriuresis and APir release by volume expansion. Plasma AP does not increase in conscious rats despite a large volume load, 30-40% of the total blood volume given in 15 min, and the natriuresis-diuresis appears to also be independent of vasopressin. On the other hand, the diuresis induced by acute volume expansion in anesthetized rats seems dependent on the elevated APir, since rats made autoimmune to AP (which are nonresponsive to exogenous AP infusions) exhibit a diuresis in conscious but not anesthetized rats. We therefore conclude that the participation of AP in volume homeostasis is more likely in pathophysiological states and that another mechanism or possibly another atrial factor mediates the diuresis-natriuresis induced by volume expansion in conscious rats. PMID- 2966402 TI - EORTC protocol 30805: a phase III trial comparing orchidectomy versus orchidectomy and cyproterone acetate and low dose stilboestrol in the management of metastatic carcinoma of the prostate. PMID- 2966404 TI - Quality of life assessment in prostatic cancer. PMID- 2966403 TI - Lessons from phase III-trials on the hormonal treatment of prostatic cancer. I: Results of EORTC-trials 30761 and 30762. PMID- 2966405 TI - ICI 176,334: a novel non-steroidal, peripherally-selective antiandrogen. PMID- 2966406 TI - LHRH-analogues therapy for metastatic prostate cancer. PMID- 2966407 TI - Advantages and disadvantages of pure antiandrogens and of antiandrogens of the cyproterone acetate-type in the treatment of prostatic cancer. PMID- 2966409 TI - Analgesia in defeated mice: evidence for mediation via central rather than pituitary or adrenal endogenous opioid peptides. AB - Mice subjected to defeat in a social conflict paradigm display an analgesic response that is apparently mediated by endogenous opioids. It is blocked by naloxone and shows full cross-tolerance to and from morphine. The present study investigated the contribution of sources of endogenous opioids outside of the central nervous system, namely the pituitary and adrenal glands. Treatment known to enhance (metyrapone pretreatment), reduce (2% saline in the drinking water) or block (dexamethasone pretreatment) the release of beta-endorphin from the anterior pituitary did not affect the display of analgesia in defeated mice. Similarly, treatments known to enhance (reserpine pretreatment) or block release of enkephalins (removal of the adrenals or hexamethonium pretreatment) from the adrenal medulla also failed to influence defeat-induced analgesia in the expected manner. If anything, adrenalectomy enhanced and reserpine pretreatment suppressed the analgesic response to defeat. The data are discussed in terms of providing evidence that defeat-induced analgesia is mediated primarily by endogenous opioids released and acting within the central nervous system. PMID- 2966408 TI - A comparison between surgical orchidectomy and LH-RH analogue ('Zoladex', ICI 118,630) in the treatment of advanced prostatic carcinoma--a multi-centre clinical study. PMID- 2966410 TI - Behavioral differentiation between effects elicited at dopamine D-1 and D-2 receptors in rats with normosensitive DA receptors. AB - The interaction between D-1 and D-2 receptor agonists was investigated in three behavioral test models. In two models (circling behavior in rats with hemitransection; stereotyped behavior in naive rats) an enabling role of D-1 receptor agonists in expressing the effect of D-2 agonists was found: D-1 agonists alone were ineffective, D-2 agonists had low maximum effects, whereas the combination induced marked activity. Consistent with this, the effect of combined D-1/D-2 receptor stimulation was antagonized either by D-1 or D-2 antagonists. In contrast, the effect of D-1 and D-2 agonists and antagonists were differentiated in drug discrimination studies, using the cue induced by the D-1 agonist SK & F 38393 or by the D-2 agonist (-)-NPA versus saline. The results thus indicate a functional relevance of D-1 receptors and show that the coupling between D-1 and D-2 receptors critically depends on the specific behavior under study. PMID- 2966411 TI - Cartilaginous histology of the cleft lip nose: proving the extrinsic etiology. AB - In an effort to prove the etiology of the cleft lip nasal deformity, 23 patients with unilateral cleft lip underwent biopsy through the midportion of the columella from mucosa to mucosa. This tissue "sandwich" contained an internal control of cleft and noncleft medial crus cartilage. With the use of special stains and examination under the microscope at low, medium, and high powers, sections were evaluated on the basis of presence of abnormal chondrocytes, number of binucleate chondrocytes, number of nucleated chondrocytes, number of lacunae, perichondrial thickness, and cartilage thickness. In each specimen examined, there were no significant differences between cleft and noncleft sides, proving histologically the previous subjective observation that the deformity is extrinsic, due to distortion of the lower lateral cartilage by abnormal vectors of force. PMID- 2966412 TI - The superiorly based rectus abdominis flap: predicting and enhancing its blood supply based on an anatomic and clinical study. AB - A detailed investigation of the blood supply of the superiorly based rectus abdominis muscle flap and the transverse rectus abdominis musculocutaneous flap was done to improve the understanding of variations in flap viability and to explain the survival of the flap after internal mammary artery division and radiation. The study involved dissections of the internal mammary and superior epigastric systems, evaluation of pertinent angiograms, and impressions from observations of the vascular anatomy correlated with flap survival in over 600 clinical dissections. There is a diffuse intrathoracic collateral network involving the internal mammary system, with multiple branches and intercommunications on the same side, as well as across the midline. This enhances flap predictability and survival in some patients with internal mammary artery division or compromise. There is also a laterally based blood supply to the flap from the costomarginal artery at the costal margin which is sometimes well developed and may prevent flap compromise if preserved. PMID- 2966413 TI - Functional and structural evaluation of the vasculature of skin flaps after ischemia and reperfusion. AB - Free radicals and other toxic oxygen species play a role in the pathogenesis of ischemic organ damage. The abdominal skin flap has been used as a model to study the effects of superoxide dismutase on the survival of ischemic skin. We have evaluated the evolution of functional and structural injury to the vasculature after ischemic injury in superoxide dismutase-treated and control skin flaps. Ischemia was induced by creating abdominal skin flaps and occluding either the venous or both the venous and arterial blood supplies. Superoxide dismutase was administered immediately after the occlusion was released. At 1 hour of reflow, erythrocyte stasis, platelet deposition, neutrophil adherence, and injury to the endothelium of the large vessels and of the microvasculature were evident. The blood flow in the ischemic skin was only 3 percent of normal. Superoxide dismutase caused no change in the ultrastructure of the vasculature and a marginal decrease in vascular permeability in the ischemic skin at 1 hour of reflow. Increased fluorescent staining of the skin was evident after 24 hours of reflow in the superoxide dismutase-treated flaps. These findings indicate that injury to vascular endothelium by ischemia and reperfusion plays a role in the evolution of skin necrosis. PMID- 2966414 TI - Resolution of back pain with the wide abdominal rectus plication abdominoplasty. AB - Back pain is classified most easily into three types (1) muscle spasm, (2) herniated nucleus pulposus (HNP), and (3) facet syndrome and/or nerve compression. It is possible the wide abdominal rectus plication procedure will be beneficial to all three groups, but currently I limit the procedure to those back pain patients who are improved with a back brace or corset and those without back pain undergoing elective abdominoplasty. Contraindications would be (1) anticipating additional pregnancy, (2) back pain not relieved or improved by corset or brace, (3) severe or progressive disk signs, and (4) bladder and/or bowel dysfunction. Further detailed studies of the mechanism of action and of the biomechanical effect as evaluated by conventional radiology, CAT scan, and nuclear magnetic resonance imaging, as well as physical measurement, are in progress and will be forthcoming. PMID- 2966415 TI - Emotional disorder and its assessment within the severe head injured population. AB - This study examined the usefulness of self-report measures of emotional disorder in a relatively acute, severely head injured population (i.e. individuals who had sustained their injuries within the previous two years). The General Health Questionnaire, the Leeds Scales of Depression and Anxiety, and Visual Analogue Scales of Depression and Anxiety were administered to 39 severely head injured subjects and 35 'close others' who were required to give their opinions on how they thought the head injured subjects were feeling. Twenty head injured subjects were re-tested within a week. Results indicated that the head injured are able to reliably complete self-report scales as indicated by the 'close others' similar reports. The head injured seem to be relatively aware of their overt behaviours and emotions, at least when utilizing the General Health Questionnaire and the Leeds Scale of Depression. According to the measures of emotional disposition used in the present study, emotional disorder was found to be common. From the results it was further suggested that lack of a close confiding relationship predicts emotional disorder and may be used to identify those subjects at risk. Overall, the results provide confidence for the use of self-report scales as a preliminary approach to the identification of emotional disorder in the severely head injured. PMID- 2966416 TI - Hierarchy of response to handicaps. PMID- 2966417 TI - Endovascular stenting of veins and grafts: preliminary clinical experience. AB - Stenotic lesions of veins and bypass grafts are often difficult to dilate and have a high frequency of recurrence. In an effort to provide an endoluminal mechanical support, the new concept of transluminal vascular stenting was applied in four patients with stenoses of nonarterial vessels, including two with postoperative venous stenoses, one with a stenosed mesenteric artery graft anastomosis, and one with a long stenosis of the basilic vein distal to a hemodialysis shunt graft. All four were successfully treated with percutaneous transluminal angioplasty followed by endovascular stenting. All but one of the stented segments were patent, with no significant restenosis after a follow-up of 4 1/2-12 months. There have been previous reports of transluminal vascular stenting in the arterial system, and the preliminary results from this study suggest that endovascular stenting also may play an important role in the treatment of venous and graft stenoses. However, further follow-up and careful patient selection will have to be done to establish the long-term benefit of this new procedure. PMID- 2966418 TI - Lumbar spine: postoperative MR imaging with Gd-DTPA. AB - Thirty patients with failed back surgery syndrome were studied to evaluate the effectiveness of magnetic resonance (MR) imaging with gadolinium diethylenetriaminepentaacetic acid/dimeglumine (Gd-DTPA) in differentiating postoperative epidural fibrosis (scar) from recurrent disk herniation. Pre- and postcontrast MR images were interpreted without access to other diagnostic, surgical, or pathologic findings. Seventeen patients had surgical and pathologic correlation of the MR findings at 19 disk levels. The precontrast studies had a sensitivity, specificity, and accuracy of 100%, 71%, and 89%, respectively. The enhanced MR studies correctly depicted the character of abnormal epidural soft tissue in 17 patients at all 19 levels. Scar showed heterogeneous enhancement on the early T1-weighted spin-echo images obtained within 10 minutes after contrast material administration. Herniated disk did not show significant enhancement on the early studies but showed variable degrees of enhancement on delayed images in nine of 12 cases. Other criteria were found to be less useful than the pattern of enhancement. Results indicate that precontrast and early postcontrast T1-weighted spin-echo studies are highly accurate in separating epidural fibrosis from herniated disk. PMID- 2966419 TI - Close-arterial administration of the thromboxane mimetic U-46619 induces damage to the rat gastric mucosa. AB - The pro-ulcerogenic actions of the thromboxane mimetic, U-46619 on the rat gastric mucosa have been investigated, utilizing a novel technique which allows administration directly into the left gastric artery. Local intra-arterial infusion of U-46619 (100-500 ng/kg/min for 10 min) induced dose-dependent macroscopic damage in both the corpus and antral regions, characterized as vasocongestion, disruption and haemorrhage, with deep penetrating ulcers in the antral mucosa. Vascular congestion, epithelial cell and glandular disruption was observed histologically in both corpus and antral regions. Local intra-arterial infusion of lower doses of U-46619 (25-100 ng/kg/min) significantly disrupted the mucosa in the presence of 10% ethanol in a concentration which itself did not induce macroscopic damage. The damaging actions of U-46619 were substantially reduced by pretreatment with the thromboxane-receptor antagonist, BM 13,177 (5mg/kg i.v.) or 16,16-dimethyl PGE2 (5 micrograms/kg s.c.). These findings support the role of endogenous thromboxane A2 as a local mediator of gastric injury. PMID- 2966420 TI - [Dermatitis exfoliativa caused by carbamazepine]. PMID- 2966421 TI - [Regional arterial chemotherapy of hepatic metastases secondary to colorectal carcinoma]. PMID- 2966422 TI - [Risk factors in the closure of lateral colostomies]. PMID- 2966423 TI - [Effect of streptozotocin at the insulin pre-receptor, receptor and post-receptor level in adipocytes of rats]. AB - The relationship among plasma insulin disappearance, insulin binding to specific receptors in fat cells and antilipolytic insulin activity in streptozotocin diabetic rats has been studied. Male Wistar rats were injected streptozotocin (65 mg/kg body weight) or saline by cardiac puncture. Decreased insulin levels and increased insulin degradation together with an increase in insulin binding were found in diabetic rats. The increase in insulin binding was related to an increase in the number of insulin receptors rather than to a change in receptor affinity. These findings at the pre-receptor and receptor levels could be correlated with an increase in antilipolytic insulin activity. However our results suggest that the mechanism for insulin action occurred through a potentiation of the norepinephrine lipolytic activity. PMID- 2966424 TI - [Questions regarding demarcation in occupational therapy]. AB - Work therapy (Arbeitstherapie) has become an intermediate link between medical and vocational rehabilitation. It is distinct from insurable employment primarily in that it is guided by a medical treatment plan directed at the rehabilitative needs of the patient. Delimitation problems as against insurable employment mostly occur due to the requirement that the work therapy setting should be realistic. This entails a risk of abuse. Where work therapy is provided in practice, greater note should be taken of today's degree of differentiation in vocational rehabilitation, which has made available a great many avenues so that direct provision of vocational measures to the patient may be more meaningful. Preparatory vocational rehabilitation services may be considered if the patient's working capacity needs to be improved and is amenable to goal-directed enhancement. It is not necessary that this should occur in courses. It, rather, can also be done through individual measures. And it is moreover possible to include general education goals in an occupation-related manner. PMID- 2966425 TI - [Promoting exercise and sports in vocational rehabilitation of handicapped patients--a neglected aspect of rehabilitation?]. AB - Directed primarily at preventive and compensatory objectives, sports activities in the framework of vocational education are intended to enhance essentially the physical and psychic well-being of the young people in later employment. They even involve goals that are pertinent to occupational qualification. The reality of motor promotion of disabled young trainees through sports, however, does look unfavorable. Physical-motor deficits in the trainees, explained in terms of their particular biography, make up for an additional, acquired problem that can have far-reaching implications for their occupational future. Difficulties in view of systematic promotion, as well as possibilities and initial efforts towards improvement, are dealt with on the example of the vocational education centre for physically disabled young people of the Annastift, Hannover. Intensive motor promotion in more severely disabled young people during their vocational training has entailed noticably favourable results relative to coordination and physical fitness. Besides its effect of general activation, comprehensive, holistic sports motor training above all seems to have a generalized effect on everyday and work related tasks (such as gripping, walking). PMID- 2966426 TI - [Sports in vocational schools in an occupational training center for handicapped patients--prerequisites, responsibilities and organization]. AB - Sports activities in the vocational education centres are viewed as an indispensable rehabilitation measure. Its contents and organizational structure however is to a large extent left to the individual centres. Implementation of effective sports is endangered by the lack of homogeneity in the young trainee population as regards type and severity of the disability, age, maturity. After setting out the present situation and the function of sports in a vocational education centre for physically disabled young people, a school course system is presented that, by achieving a relatively homogenous composition of the sports groups, enables sport-oriented instruction to be realized. In this course system, the young people are allocated to the various courses through sport selection and counselling, based on the criteria of inclination and suitability. The range of sports offered also includes sports for the very severely handicapped, which are described briefly. PMID- 2966427 TI - [Comparison of Japan and USA in the treatment of ischemic heart disease]. PMID- 2966428 TI - [Fluctuation of atrial natriuretic polypeptide during the acute exacerbation of chronic respiratory failure]. PMID- 2966430 TI - [Genetic epidemiology of Huntington chorea in Chile]. PMID- 2966429 TI - [Paroxysmal atrial fibrillation, a cause of systemic embolism?]. PMID- 2966431 TI - [The history of the development of rheumatologic services in the Chuvash A.S.S.R]. PMID- 2966432 TI - [Current status and ways of improving the specialized rheumatologic services for the population of the Georgian S.S.R]. PMID- 2966433 TI - Effect of pentagastrin and cimetidine on gastric blood flow measured by laser Doppler flowmetry. AB - Endoscopic laser Doppler flowmetry has been used to study the effect of the secretagogue pentagastrin and the H2-receptor antagonist cimetidine on gastric blood flow in 24 healthy subjects. The subcutaneous injection of pentagastrin, 0.6 micrograms/kg body weight, caused a significant increase in gastric acid secretion. This increase did not provoke any significant change in gastric blood flow, measured in seven defined areas of the oesophagus and stomach. Furthermore, no significant correlation was found between the level of basal and maximal acid secretion and blood flow. The infusion of 200 mg cimetidine caused a significant reduction of gastric blood flow in five of the seven investigated areas (P less than 0.05 and P less than 0.01). Cimetidine had no effect on blood pressure or skin blood flow, measured by laser Doppler flowmetry. The study was performed without previous stimulation of acid secretion, and blood flow reduction was measured in both the corpus and the antrum of the stomach. The effect of cimetidine is therefore hardly a result of the acid-reducing effect of the drug. In this study no significant relationship could be shown between the gastric wall blood flow and acid secretion. PMID- 2966434 TI - Self-reactive delayed type hypersensitivity induced in mice by syngeneic lymphoblasts. III. Immunological characterization of the small and large antigens of the blast cells. AB - X-irradiated or normal A mice injected with syngeneic concanavalin A-induced lymphoblasts (syn-Con A blasts) developed inflammatory responses in their footpads 24 to 72 h after the injection of syngeneic lipopolysaccharide-induced lymphoblasts (syn-LPS blasts) into these tissues. This response was designated syngeneic delayed type hypersensitivity (syn-DTH). The Con A blast extracts contain small (apparent MW of 6000-7000) and large (apparent MW of 160,000 175,000) syn-DTH-stimulating antigens, which are found in the total volume (low molecular weight fraction) and the void volume (high molecular weight fraction), respectively, of AcA 44 gel filtrations of this extract. The small and large antigens exhibit different immunological properties. The small antigen of A mouse lymphoblasts induced syn-DTH in X-irradiated (250 rad) mice but not in normal mice, and this immunological activity was elicited with syngeneic but not allogeneic lymphoblasts. The syn-DTH induced with the small antigen was inhibited by Lyt-1+2+, I-Jk+ suppressor T cells or a factor extracted from these cells. In contrast to the small antigen, the large antigen of A mouse lymphoblasts induced syn-DTH in both normal and X-irradiated mice, and this immunological activity was elicited by both syngeneic and allogeneic LPS lymphoblasts. The small and large antigens do not immunologically cross-react, but their immunogenicity is not affected by ultraviolet irradiation, indicating that the immune response against both of them is relatively class II-independent. The possibility that the cellular autoanti-lymphoblast response observed in our studies is in fact a mechanism that down-regulates the lymphoblast activity and thus suppresses the immune response is discussed. PMID- 2966435 TI - Differences between primed allogeneic T-cell responses and the primary mixed leucocyte reaction. Primed T cells become independent of the blocking effects of monoclonal antibodies against IL-1 beta and the CD5, CD11a (LFA-1), and CD11c (p 150,95) molecules. AB - Recent investigations have demonstrated that the primary mixed lymphocyte reaction (MLR) is dependent on certain accessory molecules, e.g. CD4 and LFA-1. We have compared the requirements of the primary MLR and the responses of alloreactive, primed lymphocytes (PL) by inhibition studies using monoclonal antibodies (MoAb) directed against (i) adhesion molecules belonging to the CD11 cluster of leucocyte antigens (CD11a, LFA-1; CD11b, MAC1 = CR3; and CD11c, p 150,95); (ii) various T cell-related antigens (CD2, CD4, CD5 and CD8); and (iii) recombinant IL-1 beta. The CD5-, CD11a- and CD11c-reactive MoAb significantly inhibited the primary MLR (inhibition = 25%, P less than or equal to 0.01; 48%, P less than or equal to 0.01 and 13%, P less than or equal to 0.05, respectively) but these MoAb did not inhibit the primed lymphocyte reaction (PLR). The CD11b reactive MoAb had no significant influence on either of the responses. CD2- and CD4- reactive MoAb significantly inhibited both primary MLR (greater than 80%, P less than or equal to 0.01) and to a lesser extent the PLR (40-65%, P less than or equal to 0.01). A MoAb reactive with IL-1 beta inhibited the primary MLR (38%, P less than 0.01) and the purified protein derivative (PPD) induced lymphocyte transformation response (42%, P less than or equal to 0.01) of peripheral blood mononuclear cells (PBMC), whereas primed allogeneic responses to PBMC and Epstein Barr virus (EBV) cell lines were unaffected by this MoAb. In addition, preliminary data indicated that PL seemed neither to bind exogenous IL-1 (as opposed to CD4+ PBMC) nor to possess membrane-bound IL-1. The differences between 'virgin' and primed, allogeneic T-cell responses indicate that profound changes in the functional capability of the responding T-cell population take place during the bulk expansion. The results indicate that during repeated priming with alloantigen and bulk expansion, the proliferative response of T lymphocytes becomes independent of (i) the interaction with the CD11 adhesion molecule(s), (ii) the CD5 molecule, and (iii) the cytokine IL-1 beta. PMID- 2966436 TI - [Lyme carditis]. AB - In 1980 a large series of patients with Lyme carditis was first described. The most frequent symptom was atrioventricular block, more unusual were other arrhythmias, nonspecific ECG alterations, cardiomegaly and pericarditis. We report the case of a 51-year-old man with Lyme carditis which presented as atrial fibrillation and atrioventricular block 2 degrees. Gallium-67 scan was negative. The arrhythmias ceased during therapy with doxycycline. The clinical findings, diagnosis and therapy of Lyme carditis are discussed. PMID- 2966437 TI - Immunotherapy of the nonobese diabetic mouse: treatment with an antibody to T helper lymphocytes. AB - Spontaneous diabetes mellitus was blocked in nonobese diabetic mice by treatment with a monoclonal antibody against the L3T4 determinant present on the surface of T-helper lymphocytes. Sustained treatment with the monoclonal antibody led to cessation of the lymphocytic infiltration associated with the destruction of the insulin-producing beta cells. Moreover, the mice remained normoglycemic after the antibody therapy was stopped. These studies indicate that immunotherapy with monoclonal antibodies to the lymphocyte subset may not only halt the progression of diabetes, but may lead to long-term reversal of the disease after therapy has ended. PMID- 2966438 TI - Anti-acne drug poses dilemma for FDA. PMID- 2966440 TI - [Use of physostigmine in anesthesiology]. PMID- 2966439 TI - [Gastrodyne, a glycopyrrolate in anesthesiology practice]. PMID- 2966441 TI - [A case of Sjogren's syndrome associated with palmoplantar pustulosis]. PMID- 2966443 TI - Clinical observations and lessons learned in the treatment of patients with thoracoabdominal aortic aneurysms. AB - Thoracoabdominal aortic aneurysms can be repaired successfully with acceptable rates of morbidity and mortality. Twenty-three men and seven women (an average age of 67 years) underwent 23 elective and seven emergency operations. Pulmonary complications were the most common, but renal insufficiency and paraplegia were the most serious postoperative problems. The average time of suprarenal aortic occlusion was 47 minutes, but neither renal insufficiency nor paraplegia was directly related to suprarenal clamp time. Four deaths occurred after elective procedures, two from postoperative bleeding (one death from a technical error) and two deaths from multisystem organ failure. Four late deaths were caused by myocardial infarction. The remaining patients are alive at two to 79 months after infarction. DeBakey's technique (multiple sidearm grafts from the main aortic graft) was used in the first three procedures, and the graft inclusion technique of Crawford, in the remainder. The graft inclusion technique reduced operating time and loss of blood by 50 per cent and intraoperative fluid requirements by 33 per cent. Hypothermia was minimized by extraperitoneal, rather than intraperitoneal, abdominal aortic exposure, heated ventilation and warmed intravenous fluids. Selective renal cooling was performed by catheter perfusion of the renal arteries. Extreme care must be taken in making openings and attaching grafts to visceral arteries to avoid troublesome hemorrhage. In contrast with patients with infrarenal aneurysms, those with thoracoabdominal aneurysms require prolonged ventilatory support and have considerably higher fluid requirements. Precise surgical technique is mandatory. PMID- 2966442 TI - The systemic activation of platelets by Dacron grafts. AB - Dacron (polyester fiber), a stimulus to platelet aggregation in vitro, accumulates platelets to a greater extent in vivo than autogenous artery, polytetrafluoroethylene (PTFE) or human umbilical vein (HUV). We conducted a series of experiments using the ex vivo shunt in the baboon to determine whether or not systemic activation of platelet function was produced by a Dacron graft. Two 5 centimeter segments of 4 millimeter internal diameter graft materials were placed in series in the ex vivo shunt perfused at 25 milliliters per minute flow rate for two and one-half hours. Deposition of autologous Indium 111 labeled platelets was monitored. The ex vivo shunt procedures were divided into two groups, both with PTFE as the proximal graft: one with a distal Dacron graft (n = 21), the second with PTFE or HUV distally (n = 17). In this study, an increase in platelet deposition on the proximal PTFE graft represents systemic platelet activation caused by the distal graft. Increased platelet deposition on PTFE was noted at all time points in the presence of a Dacron graft (p less than 0.05). This property of Dacron has important clinical implications, potentially accelerating the progression of vascular disease, increasing the failure rate of composite grafts and subsequent arterial reconstruction. PMID- 2966444 TI - [Characteristics of the course of non-fatal ischemic heart disease (data of a prospective study of the open population of men over 40]. AB - A total of 2096 males born in 1916-1935, residing by 1976 in the Petrogradsky District of Leningrad, were twice examined (with an interval of 7.6 yrs) using epidemiological methods. This group was equal to 53.6% of the persons subjected to the first screening. Data on the dead and those who had not come to the second screening, were not analyzed. The total frequency of nonfatal CHD from 1976 up to 1984 raised from 12.6 to 19.2%. As was shown by the assessment of a WHO questionnaire with relation to angina of effort and an analysis of ECG at rest, CHD was undetectable during the second screening in some of the examinees with CHD detected during the first screening. Further on the angina syndrome was not reaffirmed in 52.3% of the patients with previously diagnosed angina, "ischemic" ECG changes were not recorded in 37.1% of the persons with previously detected ischemic codes. Positive ECG time course was noted in 52.1% of the persons with ECG changes typical of CHD detected during the first screening. In spite of a marked tendency to an increase in the total CHD frequency a long-term observation demonstrated a possibility of a favorable time course of chronic CHD and remission in this disease. PMID- 2966445 TI - [Nutrition, the prevalence of ischemic heart disease and risk factors of its development in men aged 40-49 years in the city of Frunze in relation to ethnic groups]. AB - An epidemiological survey was conducted among men aged 40 to 59 to study interrelationships of the nutritional patterns in 2 ethnic groups of population and the prevalence of CHD and risk factors of its development. The nutritional habits of the indigenous Kirghiz population in the city of Frunze differed significantly from those of the alien population (Russians, Ukrainians, etc.) and were characterized by a higher consumption of proteins and complex carbohydrates. Nutrition of the alien population included mostly simple carbohydrates: sugar, fat and monounsaturated fatty acids. Coronary heart disease, arterial hypertension, smoking, excess body mass, low physical activity were equally common among the Kirghiz and Europeans. Hypercholesterinemia was noted more frequently in Europeans whereas hypertriglyceridemia was more common among the Kirghiz. PMID- 2966446 TI - [Pathology of the external respiratory system (data of mass screening) and general risk factors in non-infectious diseases]. PMID- 2966447 TI - [Effectiveness of complex sanatorium-health resort rehabilitation of patients with chronic nonspecific lung diseases on the southern coast of Crimea]. AB - The paper is concerned with the results of a study of efficacy of rehabilitation of 1174 patients with chronic nonspecific pulmonary diseases in sanatoria. On the basis of clinical criteria and the results of bicycle ergometry 4 clinicofunctional groups of patients were defined. A tactical approach to the differentiated use of different types of therapeutic, physical and psychic rehabilitation was worked out for these groups. An index method for assessment of the efficacy of rehabilitation based on 5 groups of symptoms was proposed. Therapeutic efficacy in different clinicofunctional groups varied from 89.6% in the 1st group to 71.8% in the 4th group; it was higher among the patients with chronic nonobstructive bronchitis (from 89 to 98%) than among the patients with chronic obstructive bronchitis and bronchial asthma (from 70 to 80%). PMID- 2966448 TI - [Clinico-genetic characteristics of hereditary nephritis in different populations]. AB - Some data are presented on the clinical features of a course of hereditary nephritis in persons of 13 nationalities residing in the central zone of the RSFSR, in Central Asia and East Slovakia (the Czechoslovak Socialist Republic). At least 2 types of hereditary nephritis transmission (the dominant x-chromosome linked one was more common than the autosomal-dominant one) not differing in their clinical course were revealed. Basing on a study of the clinical features of a course of disease in 123 autobred and 52 inbred families a more severe course of nephritis was observed in children from the inbred families. PMID- 2966449 TI - The preparticipation sports examination in Special Olympics athletes. PMID- 2966450 TI - The frequency of type I heterozygous protein S and protein C deficiency in 141 unrelated young patients with venous thrombosis. AB - The frequency of heterozygous protein C and protein S deficiency, detected by measuring total plasma antigen, in a group (n = 141) of young unrelated patients (less than 45 years old) with venous thrombotic disease was studied and compared to that of antithrombin III, fibrinogen, and plasminogen deficiencies. Among 91 patients not receiving oral anticoagulants, six had low protein S antigen levels and one had a low protein C antigen level. Among 50 patients receiving oral anticoagulant therapy, abnormally low ratios of protein S or C to other vitamin K dependent factors were presented by one patient for protein S and five for protein C. Thus, heterozygous Type I protein S deficiency appeared in seven of 141 patients (5%) and heterozygous Type I protein C deficiency in six of 141 patients (4%). Eleven of thirteen deficient patients had recurrent venous thrombosis. In this group of 141 patients, 1% had an identifiable fibrinogen abnormality, 2% a plasminogen abnormality, and 3% an antithrombin III deficiency. Thus, among the known plasma protein deficiencies associated with venous thrombosis, protein S and protein C deficiencies (9%) emerge as the leading identifiable associated abnormalities. PMID- 2966451 TI - Plasma protein S activity measured using Protac, a snake venom derived activator of protein C. AB - Functional activity of protein S, a cofactor of activated protein C-dependent inhibition of blood coagulation, in human plasma was measured by using Protac, a snake venom derived activator of protein C. This assay appeared to be specific for protein S, because 1) the activated partial thromboplastin time of protein S depleted plasma depended on the purified protein S added in the presence of Protac; and 2) the level of protein C in plasma sample (0 to 10 micrograms/ml) had no influence on the clotting time. The cofactor activity of protein S in the plasma of normal men (n = 16) and women (n = 14) was 99.4 +/- 23.8% and 98.6 +/- 24.5% respectively. The protein S activity in the plasma of pregnant women at pre and post-partum (n = 14), and that in the plasma of patients under warfarin therapy (n = 20) were 46.2 +/- 18.9%, 45.8 +/- 19.6% and 24.0 +/- 15.7%, respectively. In these plasmas, the levels of protein S activity were lower than those of total protein S antigen, but were similar to those of free protein S antigen. In 16 patients out of two families with congenital protein S deficiency, the protein S activity, the free antigen and the total antigen were 9.4 +/- 6.9%, 13.3 +/- 4.6% and 57.4 +/- 20.7%, respectively. There was no significant relationship between the level of protein S activity and that of a complemental C4b-binding protein antigen in any of these patients. PMID- 2966453 TI - [Laparoscopy in infertility. A comparison of findings and treatment]. PMID- 2966454 TI - [Thyroid function in adults with Down's syndrome. A study from an institution]. PMID- 2966452 TI - Physiological coagulation inhibitors (protein S, protein C and antithrombin III) in severe preeclamptic states and in users of oral contraceptives. AB - Protein C, protein S and antithrombin III were evaluated in normal pregnancy, severe preeclampsia and chronic hypertension with superimposed severe preeclampsia. The same study was performed on a group of 10 normal women using oral contraceptives. In normal pregnancy a significant decrease in the level of free and total PS was observed in the 2nd trimester of pregnancy and was sustained throughout the remaining months. No significant changes in the levels of protein C and antithrombin III were observed during normal pregnancy. In preeclamptic states a significant decrease in protein C was observed. It was more evident in severe preeclampsia when compared with the normal pregnancy group at similar gestational age. No statistically significant differences in protein S were found when the normal and pathological groups were compared. Antithrombin III decreased only slightly in the severe preeclamptic group. The decrease in protein C and antithrombin III levels in severe preeclampsia could be related with the microthrombotic state that these patients may present. However, the role played by protein S, which decreases during normal pregnancy and in preeclampsia, is not clear. A decrease in the level of total protein S was observed in the group of women using oral contraceptives. No significant changes in protein C and antithrombin III levels were observed in this group. PMID- 2966455 TI - [The eruption time and status of the deciduous teeth and bite of young children living in rural districts of Georgia]. PMID- 2966456 TI - [Results of a dental examination of children, native inhabitants of the Nenets Autonomous Okrug]. PMID- 2966457 TI - [Academic, scientific and practical cooperation as an efficient form of work to introduce the comprehensive prevention of stomatological diseases in children in rural localities]. PMID- 2966458 TI - [Teaching of pedodontics and the prevention of stomatological diseases in children]. PMID- 2966459 TI - [The role of the student olympiad in stimulating academic activities]. PMID- 2966460 TI - [Forensic medicine expertise on hair]. PMID- 2966461 TI - [85th anniversary of the Department of Forensic Medicine of the Pavlov Leningrad Medical Institute of the Order of the Red Banner of Labor]. PMID- 2966462 TI - The substitution of carrier priming of helper function in the common American newt, Notophthalmus viridescens by lectins and human lymphokines. AB - There is no clear evidence that helper function is thymus dependent in the Common American newt, Notophthalmus viridescens. Here we test the capacity of concanavalin A, wheat germ agglutinin, human rIL-1 and rIL-2, reagents which stimulate T cell activities in other species, to substitute for carrier priming in the newt. Cytofluorimetric analyses have been used to demonstrate specific IL 2 receptor binding sites on newt splenocytes. Competitive pre-binding with rIL-2 tested whether anti-IL-2 receptor antibody binding sites would bind rIL-2. While Con A can substitute for carrier priming in the newt only when it is presented on Sepharose or agarose particles, wheat germ agglutinin cannot, even when it is injected in particulate form. Additionally, human rIL-1 can serve as an effective substitute for carrier priming, but rIL-2 cannot. The cytofluorimetric data are in agreement with the functional data in that they suggest that human rIL-2 may not bind newt splenocytes. Our data which show shared lectin specificities with T cell regulated helper function in another amphibian species are consistant with the possibility that T-like cells are responsible for helper function in this species. PMID- 2966463 TI - Thymic influence on the T-lymphocyte self MHC repertoire. I. Helper T-lymphocyte precursors. AB - We measured the frequencies of helper T-cell precursors in spleens of allogeneic thymus-grafted nude mice to determine whether allogeneic thymus engraftment resulted in clonal deletion of helper T-cells reactive to thymic major histocompatibility complex alloantigens, thereby producing tolerance to the thymic alloantigens. C3H thymus-grafted nudes had nearly normal numbers of C3H reactive helper T-cell precursors, whereas C57BL/6 thymus-grafted nudes had significantly reduced numbers of C57BL/6-reactive helper T-cell precursors. Additional evidence suggested that tolerance was not due to a paucity of helper T cell precursors: a) there was no correlation between the helper T-cell precursor frequency and the ability to mount cytotoxic responses against the thymic alloantigens, and b) exogenous helper factors did not break cytotoxic T lymphocyte tolerance to thymic alloantigens. Thus, we conclude that immune tolerance resulting from engraftment of allogeneic thymic tissue is not necessarily due to clonal deletion of specific helper T-cell precursors. PMID- 2966464 TI - Cloning of human CD3+ T cells and CD3- natural killer cells and requirements for long-term expansion. PMID- 2966465 TI - Construction of rat-mouse T hybridoma clones from rat kidney allograft infiltrating cells with helper activity. PMID- 2966467 TI - A non-cytotoxic T cell clone from a human kidney graft infiltrate inhibits factor VIII synthesis by donor kidney endothelial cells. PMID- 2966466 TI - Induction of "tolerance" in monoclonal human helper T lymphocyte populations during culture under conditions of chronic alloantigenic stimulation. PMID- 2966468 TI - Modulation of graft-versus-host associated cytotoxic T cell suppression by preimmunization of the recipient. PMID- 2966469 TI - Different functional activities of T cell lines isolated from a rejected renal allograft and peripheral blood lymphocytes of renal transplant recipients. PMID- 2966470 TI - T lymphocyte infiltrates in inflammatory synovia are oligoclonal. PMID- 2966471 TI - Human CD3+4-8- TCR alpha beta- gamma+ blood T cells, their lytic potential against tumor cells, and their recognition using WT31 monoclonal antibody. PMID- 2966472 TI - Evidence for a cyclosporine-resistant pathway of T cell activation triggered via the CD3/T cell receptor complex. PMID- 2966473 TI - In vitro effects of cyclosporines A and G on activation of an autoimmune T cell line. PMID- 2966475 TI - In vitro effects of cyclosporine A on allogeneic and autologous lymphocyte stimulation: influence of HLA phenotypes. PMID- 2966474 TI - Immunosuppressive effect of cyclosporine metabolites from human bile on alloreactive T cells. PMID- 2966476 TI - The immunobiology of experimental lung transplantation with cyclosporine immunosuppression: cytolytic T lymphocytes and delayed type hypersensitivity in rejecting and tolerant recipients. PMID- 2966477 TI - Effects of cyclosporine A on T lymphocytes and accessory cells from human blood. PMID- 2966478 TI - Synergistic effects of cyclosporine and rabbit antithymocyte sera on suppression of the one-way allogeneic mixed lymphocyte response. PMID- 2966479 TI - Additive effect of cyclosporine and verapamil on the inhibition of activation and function of human peripheral blood mononuclear cells. PMID- 2966480 TI - Important role of cyclosporine for the induction of immunologic tolerance in adult hosts. PMID- 2966481 TI - Comparison of binding to peripheral blood lymphocytes between active and inactive derivatives of cyclosporine. PMID- 2966482 TI - A study of the correlation between cyclophilin binding and in vitro immunosuppressive activity of cyclosporine A and analogues. AB - In order to establish whether CyP is the pharmacologically relevant CsA receptor, the CyP binding v immunosuppressive activity was measured for an extensive, structurally varied group of CsA analogues. Overall, CyP binding was found to parallel immunosuppressive activity. Other than MeAla6-CsA, the few exceptions to the correlation could be ascribed to cellular metabolism. These results strongly implicate CyP or a related protein in the mechanism of action of cyclosporine. PMID- 2966483 TI - Cyclosporine pharmacokinetics and pharmacodynamics after oral administration in the rat. PMID- 2966484 TI - In vivo fluorescence microscopy reveals cyclosporine G to be less nephrotoxic than cyclosporine A. PMID- 2966487 TI - Treatment with clodronate in patients with hypercalcemia secondary to malignancy. AB - Dichloromethylene bisphosphonate (clodronate, Cl2MDP) is a synthetic analogue to pyrophosphate, which inhibits increased bone resorption. This drug was given to 12 patients with hypercalcemia secondary to advanced malignant disease. Clodronate in a daily dose of 1.6 to 3.2 g generally caused a return of the serum calcium values to normal within 5-10 days with a concomitant improvement of symptoms related to the hypercalcemia. Side effects were few. Thus, clodronate appears to be a valuable adjunct for the medical management of patients with malignancy-associated hypercalcemia. PMID- 2966485 TI - Cyclosporine A acts as a potent co-stimulator in up-regulation of IgE antibody synthesis and expression of Fc receptor for IgE--a unifying hypothesis of the cellular mechanisms of cyclosporine A. PMID- 2966486 TI - Association of HLA and other genetic markers in South Indian patients with pulmonary tuberculosis. AB - Histocompatibility antigens (A, B & C loci) and 23 other single gene characters were studied in 204 pulmonary tuberculosis patients belonging to a single endogamous group in South India. None of the previously reported associations with HLA antigens was confirmed, nor any new one found. The blood O and Rh negative associations were also not confirmed, although a new association with the Jk blood group system appears possible. Of particular interest is the association with the phosphoglucomutase (PGM1) system, which parallels that found in a different population located some 1000 km away. Relative risks were calculated to measure the resistance of individuals with the PGM1*2+ allele. PMID- 2966488 TI - [State of the left atrium in the open atrioventricular canal]. PMID- 2966489 TI - The histidine-221 to tyrosine substitution in v-mos abolishes its biological function and its protein kinase activity. AB - The viral mos gene encodes a cytoplasmic transforming protein termed p37mos. Evidence gathered from a number of experimental approaches is consistent with p37mos having a serine/threonine protein kinase activity. To gain further understanding of the p37mos-associated biochemical activity, we constructed a mutation in the v-mos gene by oligonucleotide-directed mutagenesis yielding a histidine to tyrosine substitution at residue 221 in p37mos. Based upon nucleotide sequences, the histidine residue at the corresponding position is conserved in all the serine/threonine protein kinases from yeast to man, and is absent in protein-tyrosine kinases. The mutant p37mos (Tyr-221) was expressed in yeast and assayed for kinase activity. The mutant protein was inactive as judged by a loss of autophosphorylation activity in vitro, thus providing further support for the conclusion that p37mos is a protein kinase. When the mutant v-mos gene was introduced into a retroviral vector, pDD102, and assayed for focus forming ability on NIH/3T3 cells, it was found to be inactive at both 37 and 30 degrees. In contrast, the wild-type v-mos had transforming activity at both temperatures. These results extend our earlier findings on the correlation between transforming ability and protein kinase activity. A histidine to tyrosine substitution at the corresponding position of the v-mos protein and the yeast CDC28 gene product causes a similar effect on the kinase activity. Therefore, this residue and/or the sequence near the N-terminal side of the conserved predicted phosphate transfer domain, near the middle of the complete catalytic domain, might be specifically involved in the catalytic activity of serine/threonine protein kinases in general. PMID- 2966490 TI - Isolation of mutations of the phage Mu ner gene. AB - A method was devised which allows the easy detection of mutations within the ner gene of Mu DNA. This method is based upon the observation that a transcriptional gene A-galK fusion containing the complete ner gene and the cts62 allele does not express the galK gene in an Escherichia coli strain lacking functional integration host factor under inducing conditions (white colonies on MacConkey galactose plates at 42 degrees) In contrast, a gene ner-galK fusion which lacks part of the ner gene exhibits GalK activity (red colonies) on MacConkey galactose plates at 42 degrees. After mutagenesis of a plasmid carrying a transcriptional gene A-galK fusion, putative ner mutants could be identified on indicator plates. Cloning experiments locate the mutation(s) to the right of the HindIII site which is situated within the early promoter of Mu DNA. One of the mutants was sequenced and revealed two substitutions: one within the-10 region of the early promoter, and another near the end of the ner gene. The former lesion was shown to be pleiotropic. PMID- 2966491 TI - [Effect of vitamin A deficiency on ATPase activity and the transport functions of erythrocyte membranes]. AB - Influence of vitamin A deficiency on ATPase activity in erythrocytes was studied experimentally. It was shown that activity of Mg2+, Na+, K+-ATPase increased, while that of Na+, K+-ATPase decreased under such conditions. The assay of K+ and Na+ in erythrocytes and plasma revealed diminished K+ content in erythrocytes and its elevated level in plasma, Na+ concentration in erythrocytes rose. The author considers that disorders of protein biosynthesis (at the level of translation) in hypovitaminosis A are responsible for the results obtained. PMID- 2966492 TI - [Benzo(a)pyrene content of grain and the products of its processing]. AB - The content of benz(a)pyrene (BP) was studied in the grain, different sorts of flour and in talkan, a Kazakh national food product, prepared from fried grain. It was found that the grain contained 0.33 micrograms/kg of BP, flour--33-58% of that in the grain. The carcinogen content in flour varied with its kind: the highest BP concentration was detected in rye flour, the lowest--in high-quality wheat flour. In talkan, prepared from fried wheat, the content of BP was twice as much compared to the non-fried wheat, due to the intensity of frying. A man, who makes use of talkan during his life, may receive a significant amount of BP which can be conducive to raising the incidence of tumors of the digestive organs, especially of the esophagus. PMID- 2966493 TI - [Improved sanitary bacteriological control of dairy production]. AB - Investigation of bacterial contamination in curds and sour cream, and experimental studies on survival rate of a number of pathogenic and opportunistic microorganisms in these products have permitted standardization of the products with respect to coliform bacteria. A permissible level has been determined for bacterial contamination of curds and sour cream. PMID- 2966494 TI - Industrial accidents--worker compensation laws and the medical response. PMID- 2966495 TI - The working environment of the working poor: an analysis based on workers' compensation claims, census data and known risk factors. AB - Analysis of 1980 U.S. Census data reveals that a sizeable percentage of adult females employed full-time (working more than 2080 hours in 1979) worked in female-dominated service, sales and factory occupations and had family incomes which placed them in poverty or impoverishment status. Workers' compensation data on women in these occupations, drawn from the 27-state Supplemental Data System, show that they filed approximately 250,000 claims for compensation in 1980. Analysis of the claims by nature of injury and body part affected is consistent with the published medical literature. Injury to the back is the leading complaint filed and large numbers of hand and wrist injuries are also reported. Compensation data, however, are usually reflective of acute conditions, especially traumatic injury, not of chronic illnesses and injuries. Additional data are therefore drawn from the medical and scientific literature. This review shows the presence of cancer-causing agents, infectious agents, reproductive toxins, safety hazards and social stressors in these occupations. Recommendations for future programs and needs are presented. PMID- 2966496 TI - [Percutaneous peripheral laser angioplasty: initial clinical results]. AB - Laser angioplasty has developed as a new method for the treatment of peripheral arterial occlusive disease. In 19 patients with high grade stenoses or obstructions of the superficial femoral, popliteal or posterior tibial arteries (Fontaine stage IIa-IV) percutaneous transluminal laser angioplasty was performed using a novel laser catheter system. The laser catheter itself is made of polyethylene. Its distal tip is formed ovally and marked X-ray densely. A silica fiber (core diameter 0.6 mm) for delivering the laser energy is inserted into the laser catheter. Through a sheath with hemostatic valve, laser catheter and silica fiber are introduced into the artery and then advanced to the stenosis over a guide wire. During laser angioplasty, laser catheter and silica fiber are rotated around the guide wire. We use a cw-Nd: YAG laser with a wavelength of 1064 nm. The mean degree of stenosis decreased from 92 +/- 12% before to 31 +/- 19% after laser angioplasty. By conventional balloon angioplasty a further reduction of the degree of stenosis down to 15 +/- 20% was achieved. The mean systolic Doppler ankle-arm pressure ratio improved from 0.56 +/- 0.25 before laser angioplasty to 0.89 +/- 0.24 after combined laser and balloon angioplasty. In seven patients, clinically non-significant distal embolization occurred. In no patient there was a perforation of the arterial wall. Up to now, digital subtraction angiography 3 months after laser angioplasty has been performed in five patients and showed patency of all lesions. The mean systolic Doppler ankle-arm pressure ratio was 0.84 +/- 0.20. PMID- 2966497 TI - [Morphology of the coronary arteries following percutaneous transluminal coronary angioplasty with hemopericardium]. AB - Autopsy findings in a patient with cardiac tamponade following percutaneous transluminal coronary angioplasty (PTCA) are reported. Angiography during PTCA demonstrated coronary dissection without extravasation of contrast medium at the site of angioplasty. Histologic examination of the dilated coronary segment revealed longitudinal splitting of intima and media as well as circumferential subintimal and submedial dissection. Hemorrhage into plaque, intima, media, adventitia and dissection lumina was found. Disruption of the adventitia was not present. Cardiac tamponade may occur transadventitially without complete disruption of the coronary artery and without extravasation of contrast medium. PMID- 2966498 TI - ["Current city district-oriented counseling of the elderly"--forms of approach and intervention patterns]. AB - The Federal Ministry for Youth, Family, Women and Health in the Federal Republic of Germany supported a research project on counselling processes with the elderly. The overruling aim is to enable them to live at home as long as possible. A model of preventative intervention was developed and tested by a group of gerontologists and social workers in Kassel. Every fifth inhabitant aged 60 and over in a working class district of Kassel was interviewed (n = 106) with respect to his/her living conditions and need for care. As a second step, all aged persons at risk (i.e. with low income, in social isolation, with health handicaps - as well as those of a very old age) were selected (n = 53). Two social workers contacted them at home and attempted to build up a relation of confidence with a view to active counselling measures. This paper reports some important statistical results and specifies the concepts of counselling procedures developed. They are basically biographically and help oriented, encompassing intervention in acute situations of need as well as counselling in the more restricted, traditional sense of the term. PMID- 2966500 TI - [Development of new plate tests for the detection of microbial hydrolysis of esters and oxidation of 2-hydroxycarboxylic acids]. AB - The application of the thin-agar-layer coated filter culture technique (Z. Naturforsch. 42c, 1082 (1987)) has been extended to the detection of ester hydrolysis and 2-hydroxyacid oxidation by microbial colonies. The former was performed by spraying with bromocresol purple and the latter with a salicylhydrazide reagent. Under the optimized conditions, the hydrolysis activity of more than 20 mumol/h.g-wet cells and the oxidation activity of more than 40 mumol/h.g-wet cells were usually detected directly on the filter-plate cultures of bacteria, yeasts and molds. PMID- 2966499 TI - [Stereoisomeric aromatic compounds XIX: Asymmetric reduction of 4(5) oxocarboxylic acids with baker's yeast]. AB - Asymmetric reduction of 4(5)-oxocarboxylic acids (esters) by baker's yeast and cyclization in acidic media yields optically active gamma(delta)-lactones. The evaluation of their chirality and optical purity was carried out by HPLC (HRGC) analysis of the corresponding 1,4(1,5)-diols via diastereomeric esters with (R) Mosher acid (MTPA) and (S)-O-acyllactic acids respectively. By increasing the 4(5) alkyl side chain 4R(5R) configurated gamma(delta)-lactones with high ee values are generated. PMID- 2966501 TI - 4-(2'-Carboxyphenyl)-4-oxobutyryl coenzyme A ester, an intermediate in vitamin K2 (menaquinone) biosynthesis. AB - Enzyme preparations from Mycobacterium phlei, Escherichia coli and Galium mollugo cell suspension cultures were incubated in the presence of 4-(2'-carboxyphenyl)-4 oxobutyrate (i.e. o-succinylbenzoic acid, OSB, 1), ATP, coenzyme A and Mg2+. The main product isolated from the incubation mixture was 4-(2'-carboxyphenyl)-4 oxobutyryl coenzyme A ester (2) as determined by comparison with synthetic coenzyme A esters. Synthetic and enzymically formed 4-(2'-carboxyphenyl)-4 oxobutyryl coenzyme A ester (2) was shown to be enzymically converted to an intermediate in vitamin K2 biosynthesis viz. 1,4-dihydroxy-2-naphthoic acid (5). The enzymic formation of 2-(3'-Carboxypropionyl)benzoyl coenzyme A ester (3) and 4-(2'-carboxyphenyl)-4-oxobutyryl-di-coenzyme A ester (4) was also observed. They appeared in minor amounts, however. These esters were not convertible to 1,4 dihydroxy-2-naphthoic acid (5). PMID- 2966502 TI - Twisted fibrils are a structural principle in the assembly of interstitial collagens, chordae tendineae included. AB - X-ray diffraction analysis of connective tissue samples, which contain type I and type III collagen shows that twisted collagen fibrils are a general principle of assembly. The occurrence of twisted fibrils in native wet Chordae tendineae, skin and Aorta is combined with a shorter axial periodicity of about 65 nm. This shorter D period is shown to be directly related to the tilt of the molecules, which have to be curved to build-up twisted fibrils. PMID- 2966503 TI - Recognition of HLA class II molecules by antipeptide antibodies elicited by synthetic peptides selected from regions of HLA-DP antigens. AB - Repeated immunizations of rabbits with chemically synthesized peptides from selected regions of HLA-DP histocompatibility antigens resulted in the production of specific antibodies that were then isolated from the immune sera by chromatography on Sepharose-peptide immunoadsorbents. The purified antibodies, when tested with an enzyme-linked immunosorbent assay, specifically bound to the inciting fragments; moreover, two of them recognized glycoproteins extracted by nonionic detergents from human chronic lymphocytic leukemia cells, as revealed by binding assays. The results suggest that amino acid stretches 51-61 of the alpha chain and 80-90 of the beta chain of HLA-DP histocompatibility antigens are likely exposed on the surface of the protein molecule. The specific recognition of DP regions is strongly suggested by the difference in the binding of those antibodies to soluble membrane proteins, as compared to the binding of monomorphic anti-Class II monoclonal antibodies to the same antigens. PMID- 2966504 TI - Efficacy of sustained-release radioprotective drugs in vivo. AB - In previous publications from this laboratory we suggested the use of radioprotective drugs in a sustained-release form as a practical way to cope with their high toxicity and quick metabolism and excretion. Cysteine and cysteamine, well-established radioprotectants, were used as model drugs and compressed at various concentrations (0-65%) into an insoluble tablet matrix, composed of ethylcellulose and stearic acid at various ratios and compression pressures. We demonstrated in vitro that when the release rate of the radioprotectants was measured under nitrogen, the kinetic data conformed with the Higuchi square root of time equation, indicating that the release of both drugs correlates with Higuchi's diffusional mechanism. In the present in vivo study, tablets containing cysteine or cysteamine in a slow-release matrix were implanted, into the stomachs of female rats. The rats were irradiated at various time intervals up to 12 h after implantation and their survival recorded daily. Utilizing a 1:3 ratio of ethylcellulose:stearic acid as a matrix, the protective effect of the drugs was remarkable eight hours after tablet implantation. The results reported indicate that slow-release tablet formulation is a possible method for delivering of radioprotective drugs over an extended period of time. PMID- 2966505 TI - Evidences for circadian rhythmicity in the per0 mutant of Drosophila melanogaster. AB - per0 Mutants of Drosophila melanogaster which are exposed to light-dark cycles (LD) with different Zeitgeber period (T) have a limited range of entrainment. Entrained flies show a characteristic phase relationship of activity to the LD which depends on the period of the driving cycle as expected by oscillator theory. Both facts are taken as evidence that per. possesses endogenous oscillators and that the per gene product is not concerned with central clock structures but rather might be responsible for the mutual coupling between the individual oscillators in a multioscillatory system controlling locomotor activity. PMID- 2966507 TI - [Melting point, volume and stability of blood under high pressure]. AB - The melting pressure and the specific volume of human blood was measured up to pressures of 1000 bar or more. The melting pressure curve has a slope of -138 bar K-1. The compressibility of blood is nearly twice the value of blood serum. After application of high pressure the blood exhibits some haemolysis most probably due to the effect of the steel surface of the autoclave. PMID- 2966506 TI - Conformational changes in proteins induced by low temperatures: an infrared study. AB - Infrared spectra of hemoglobin (met-hemoglobin) and myoglobin were recorded in the temperature range -110 degrees C to 30 degrees C. On cooling hydroalcoholic solutions of hemoglobin, the spectra indicate a conformational change (revealed by the appearance of a band at 1665 cm-1) compatible with the appearance of distortions in its alpha-helical structure. In the case of myoglobin smaller effects are observed. These conformational changes are entirely reversible and do not occur in frozen aqueous solutions. PMID- 2966508 TI - [Proliferative activity and regulatory subpopulations of T lymphocytes in various forms of alopecia]. PMID- 2966509 TI - [Treatment of severe forms of acne by combined administration of Roaccutan and vitamin E]. PMID- 2966510 TI - [Epidemiology and pathogenesis of occupational skin diseases in automobile factory workers]. PMID- 2966511 TI - [Experience with nizoral treatment of patients with rubromycosis]. PMID- 2966512 TI - [On the centenary of the Leningrad V.M. Tarnovskii Scientific Medical Society of Dermatologists and Venereologists]. PMID- 2966513 TI - [Limit of the reparative capacity of deciduous dental pulp]. PMID- 2966514 TI - [Use of xenon-133 clearance as a diagnostic procedure in periodontology. The determination of physiological circulation indices in the area of the maxillary and mandibular anterior teeth. Age and sex dependence]. PMID- 2966515 TI - [Precision of odontometric measurements]. PMID- 2966516 TI - [The significance of various Streptococcus mutans serotypes for the progression of caries in the dentin]. PMID- 2966517 TI - [Effect of metal ceramic restorations on the microbial composition of plaque]. PMID- 2966519 TI - [Degeneration and regeneration of the endodontium and periodontium as a replica of the pattern of phylogenetic tooth development]. PMID- 2966518 TI - [Relation between functional disorders of the temporomandibular joint and the cervical spine]. PMID- 2966520 TI - [Cranial scoliosis after unilateral carotid ligation]. PMID- 2966521 TI - [Long-term results following retroauricular full-thickness skin grafts for vestibular reconstruction]. PMID- 2966522 TI - [Salivary gland tumors in childhood and adolescence. A clinico-pathological analysis]. PMID- 2966523 TI - [Lactate dehydrogenase activity in the pulp of periodontally damaged, impacted and partially impacted teeth]. PMID- 2966524 TI - [Characteristics of a dentinal wound of the porcine tooth as viewed by scanning electron microscopy]. PMID- 2966525 TI - [Does calcium hydroxide promote the survivability of antibiotically active streptomycetes on the hard cavity floor?]. PMID- 2966526 TI - [The quality of stimulated dentinogenesis after one-time stepwise deep caries therapy of deciduous molars]. PMID- 2966527 TI - [193 cases of gastroschisis and omphalocele--postoperative results]. AB - Treatment was applied to 97 cases of omphalocele and 96 cases of gastroschisis at the Dortmund Department of Paediatric Surgery over the past 20 years. The survival rate was 122. Follow-up checks were recently applied to 56 of those former patients, after nearly ten years had elapsed from surgery. Thirty-eight of these patients were clinically examined, while questionnaires were completed for the rest. Primary closures had been performed on 50 per cent of the cases, while the defects in the other children were closed in two stages, using dura implantation or silastic pouches, or were conservatively treated. Accompanying malformations were recorded from 21 per cent of the gastroschisis cases and from 28 per cent of those with omphalocele. Overall mortality accounted for 37 per cent, with mortality in the wake of receptive operations being as high as 40 to 50 per cent, the latter rate not depending on the primary approach. One and the same risk was found to exist for conservative treatment (applicable only to closed omphalocele) and primary surgical closure, as may be seen from statistical evaluation. The highest rate of relaparotomy occurred in the wake of dura implantation and use of silastic pouches. PMID- 2966528 TI - [Epidemiological surveillance of diphtheria infection in the territory of the Azerbaijan SSR]. AB - A complex of measures carried out with a view of effecting the epidemiological surveillance of diphtheria infection has made it possible to study the level and intensity of immunity to diphtheria and tetanus, determined in the passive hemagglutination test, in the child and adult population and to reveal groups, least protected against diphtheria infection. Bacteriological studies on the biological properties of Corynebacterium diphtheriae have revealed the contamination of various groups of children with this infective agents and permitted its complete characterization with the determination of its sero-, phago- and corycinovariants. PMID- 2966529 TI - [Antigenuria in plague-infected great gerbils]. AB - In the urine of plague-infected great gerbils Yersinia pestis capsular antigen was detected by means of diagnostic preparations, both commercial and experimental (based on monoclonal antibodies). The antigen was detected in many urine samples taken from the animals over a prolonged period. The incidence and duration of antigenuria were found to be related to the survival time of great gerbils after infection and the level of antibodies in their blood. The number of animals with antigenuria markedly exceeded the number of animals from which Y. pestis was isolated, especially at a later period after infection. Examinations of urine samples from live great gerbils trapped in natural foci appears to be a method more effective in epizootiological survey than the bacteriological analysis of the animals. PMID- 2966530 TI - [The epidemic process in mumps infection]. AB - Manifestations of the epidemic process of a parotitis infection can be explained by the theory of the self regulation of parasitic systems. A characteristic feature of epidemic parotitis is the formation of epidemic foci in the absence of parotitis cases and without the penetration of the infective agent from the outside. The epidemic wave of parotitis infection decreases as the virulence of the infective agent attenuates due to its passage through persons gaining immunity in the course of the epidemic. The avirulent infective agent persists in the body of some immune carriers till a sufficient stratum of susceptible subjects accumulates in the chain of the agent circulation. The analysis points to the autonomous character of the epidemic process not only among the urban and rural population, but also among separate social and age groups of the population within one town or settlement. The findings evidence an independent formation of the epidemic variant of the infective agent in individual schools and preschool institutions. PMID- 2966531 TI - [Dynamics of the etiological structure of salmonellosis and of the biological properties of the Salmonellae isolated in a selected territory of the Moldavian SSR 1982-1985]. AB - The etiological structure of Salmonella infections and the biological properties of salmonellae, isolated in one of the regions of the Moldavian SSR where the epidemic process of Salmonella infections reflected the regularities observed on the whole territory of this republic, were studied. Changes in the predominant serovars at the period of 1982-1985 in comparison with the preceding years were shown. Salmonellae belonging to the dominating serovars were characterized by more pronounced drug resistance in comparison with other salmonellae. Some parameters of the epidemic process of Salmonella infections were found to be related to the biological properties of the causative agents of these diseases. PMID- 2966532 TI - [A new method for the surgical treatment of paralytic neurogenic bladder]. AB - As a result of topographo-anatomical examination of 30 cadavers, the authors suggest a new method for the surgical management of the paralytic form of neurogenic bladder developing in traumatic injury of the lumbar-sacral segments of the spinal cord or the cauda equina roots. The method consists in remuscularization of the neurogenic bladder by means of grafts formed from the lateral parts of the rectus abdominis muscles together with the inferior epigastric vessels supplying them. The method was used in operations of 14 patients. The period of restoration of controlled micturition ranged from 3 to 4 weeks. The volume of residual urine did not exceed 50 ml and reduced to zero with time. The outbreaks of pyelonephritis ceased. PMID- 2966533 TI - [Correlation of pain and roentgenologic findings in the spine in the aged]. PMID- 2966534 TI - [Expression and suppression of the immune response in experimental chronic cutaneous leishmaniasis]. PMID- 2966535 TI - [Independence of the delayed hypersensitivity response from protection against Leishmania mexicana pifanoi]. PMID- 2966537 TI - Warm tub bath during labor. Effects on plasma catecholamine and beta-endorphin like immunoreactivity concentrations in the infants at birth. AB - In this study 13 women bathed in a warm tub bath during the first stage labor after a strictly normal pregnancy. The control group consisted of 9 women who did not want warm tub bath. Apgar scores at 1 and 5 minutes were recorded and plasma adrenalin, noradrenalin were measured in umbilical arterial blood at birth, together with pH, pCO2 and base excess and beta-endorphin-like immunoreactivity in umbilical venous blood. No differences were found between the groups. No beneficial effect was found on infant condition of a warm tub bath during labor. PMID- 2966538 TI - Laparoscopic sterilization with Filshie Clips. AB - A study comprising 221 women sterilized laparoscopically with Filshie Clips is presented. The pregnancy rate was 1.36%, which is considered acceptable as the method was new in the Department and performed by 10 surgeons with different degrees of skill in laparoscopy. PMID- 2966536 TI - The hypothalamo-neurohypophysial system in streptozotocin-diabetic rats: ultrastructural and immunocytochemical evidence for alterations of oxytocin- and vasopressin-containing neuronal profiles. AB - The hypothalamic paraventricular and supraoptic nuclei and the neurohypophysis of rats were investigated 8 weeks after streptozotocin (STZ)-induction of type 1 diabetes. Vasopressin (VP)- and oxytocin (OT)-containing neuronal profiles were examined using the pre-embedding peroxidase-antiperoxidase technique for electron microscopy. Ultrastructural alterations were observed in the somata, dendrites and axons of VP- and OT-labelled profiles. There was no evidence, however, for alterations in the synapses associated with VP- or OT-labelled somata, dendrites and axons. The results indicate that both VP- and OT-containing neuronal profiles are involved in the ultrastructural reorganisation of the hypothalamo neurohypophysial complex during diabetic conditions. Depletion of VP- and OT containing axon profiles in the neurohypophysis may suggest increased release of both neurohypophysial hormones in STZ-induced diabetes. PMID- 2966539 TI - Incarcerated hernia following laparoscopy. PMID- 2966541 TI - Cardiac rhabdomyoma. A case report with reference to atrial natriuretic peptide. AB - An autopsy case of cardiac rhabdomyoma in a male infant is reported. Many nodules of rhabdomyoma were present in all four cardiac chambers and were microscopically composed of ovoid, glycogen-laden cells and typical "spider cells". Atrial natriuretic peptide (ANP) was immunohistochemically demonstrated in both normal myocytes and rhabdomyoma cells of both atria, but not in normal myocytes and rhabdomyoma cells of both ventricles. Ultrastructurally, atrial specific granules were present in atrial rhabdomyoma cells and normal atrial cardiocytes, and these showed ANP immunoreactivity with protein A-gold technique. It could be said that the localization and intracellular distribution of ANP in this cardiac rhabdomyoma were closely similar to those of normal human heart. With regard to the presence of ANP, cardiac rhabdomyoma cells arising in atria seemed to differ from those in ventricles, although many tumor nodules occurred in both atria and ventricles. Furthermore, it seemed that cardiac rhabdomyomas could also be divided into two parts: 1) an atrial part with ANP, and 2) a ventricular part without ANP. Therefore, this study confirms the hypothesis that cardiac rhabdomyoma is a hamartoma rather than a true neoplasm. PMID- 2966540 TI - Suppressive effect of cyclosporin A on the induction of chronic serum sickness nephritis in the rat. AB - The suppressive effect of cyclosporin A (CyA) on the development of antibody induced glomerulonephritis was analyzed in rats with serum sickness nephritis. The induction of serum sickness was established by preimmunization and subsequent continuous administration of the same antigen starting 1 month later. When CyA was given in the preimmunization stage, neither induction of glomerulonephritis nor specific antibody response was observed. In contrast, when CyA was administered in the stage of continuous sensitization, mild glomerulonephritis as well as specific antibody response developed. This phenomenon implies that the significant inhibition of the occurrence of immune complex nephritis results from the suppression by CyA of its primary response to the sensitizing antigen. PMID- 2966542 TI - Effects of a single donor-specific blood transfusion on the survival of rat cardiac allografts. AB - It is now well recognized that pre-transplant donor-specific blood transfusion (DST) has a beneficial effect on the survival of allografts. To determine the optimal interval between DST and transplantation, and to analyze the mechanisms of this effect, the survival of cardiac allografts to rats which received a single DST was examined. The cardiac allograft survival was found to be prolonged when the DST was performed 1 to 6 weeks before grafting. In addition, recipient rat sera collected 1 to 6 weeks after a single DST showed significant inhibition of a mixed lymphocyte reaction (MLR). This MLR inhibition correlated with prolongation of survival of histoincompatible rat cardiac allografts. It thus appears that a single DST given from 1 to 6 weeks before transplantation has a beneficial effect on allograft survival and that MLR inhibition may be essential for inducing the effect of transfusion on organ transplantation. PMID- 2966543 TI - Effect of phenol and halogenated phenols on energy transfer reactions of rat liver mitochondria. AB - The in vitro effects of phenol and p-halogenated phenols on mitochondrial energy transfer reactions were examined using isolated rat liver mitochondria. The relationship between physiochemical properties of phenolic compounds and their effects on mitochondria were studied. Phenol and p-halogenated phenols induced the release of K+ ions from mitochondria, suggesting a change in permeability to K+ ions. A decrease in the respiratory control index, an increase in K+ release and stimulation of latent ATPase activity were observed with these compounds in the descending order of p-iodophenol, p-bromophenol, p-chlorophenol, p fluorophenol and phenol. The concentrations of the phenolic compounds resulting in fifty percent inhibition of the respiratory control index and those resulting in fifty percent release of K+ ions significantly correlated with Hammett's substituent constant (sigma) and the hydrophobic binding constant (pi) of the compounds. PMID- 2966544 TI - Transducing chemical energy into mechanical function: a comparative view. AB - The energetics of muscle contraction can be understood in terms of the major cellular ATPases. The twitch isometric transduction efficiency is relatively constant across muscle types and species. Although many of the factors that alter the shape of the enthalpy:load relation in isotonic twitch contractions have been identified our molecular understanding is unsatisfactory and more studies are needed of mammalian muscles working closer to 37 degrees C. The thermodynamic efficiency of CB activity seems quite high, probably in excess of 70%. During maintained (tetanic) force there can be greater than a 1000 fold difference in energy usage across muscle types and there are factors that can down regulate CB activity: these factors remain to be fully identified in both skeletal and smooth muscles. The very diversity of muscle types and the different biochemical solutions that have emerged to match energy supply and demand should lead to important insights into the contractile mechanism. The corollary however also applies, it may be dangerous to take results obtained in one muscle type under a particular set of conditions, and extrapolate those findings to muscles in general. PMID- 2966545 TI - Huntington's disease. AB - Huntington's disease is characterized by striatal degeneration, leading to progressive choreoathetoid movements, dementia and behavioral changes. It is inherited as an autosomal dominant illness with complete penetrance. New technology has localized the aberrant gene, and presymptomatic and even prenatal testing may eventually be available. Although no definitive treatment currently exists, judicious use of neuroleptics, appropriate referrals and a strong therapeutic alliance with the family may be helpful. PMID- 2966546 TI - Central and regional hemodynamic effects of oral enoximone in congestive heart failure: a double-blind, placebo-controlled study. AB - Twelve patients with congestive heart failure underwent a double-blind, placebo controlled study for the purpose of examining the central and regional hemodynamic effects of first-dose (1 and 2 mg/kg) oral enoximone, a new phosphodiesterase III inhibitor. Enoximone augmented cardiac output, generally through a positive chronotropic response. Indices of left ventricular contractility, specifically stroke volume, delta P/delta t, fractional shortening rate, and the duration of the preejection period, were only modestly enhanced by enoximone. At 2 mg/kg, systemic vascular resistance fell below baseline values without affecting systemic blood pressure; these parameters were not altered by the 1 mg/kg dose. Both pulmonary artery pressure and pulmonary vascular resistance dropped below baseline and below placebo control for the 2 mg/kg dose. Enoximone at 2 mg/kg lowered right and left heart filling pressures below baseline. Examination of regional hemodynamic responses to both doses demonstrated a reduction in limb vascular resistance and an increase in limb blood flow proportional to the concomitant increase in cardiac output. Renal and hepatic-splanchnic blood flow and vascular resistances were not altered by enoximone. First-dose oral enoximone (1 and 2 mg/kg) alters hemodynamics in heart failure by predominant vasodilatation, particularly of limb-musculoskeletal and pulmonary vascular beds, some positive chronotropism, and modest positive inotropism. PMID- 2966547 TI - Atrial natriuretic peptide in congestive heart failure: postural changes and reset with chronic captopril therapy. AB - The response of atrial natriuretic peptide (ANP) to standing and the acute and chronic administration of the vasodilator, captopril, were examined in 15 patients with chronic congestive heart failure. In the supine position, ANP was increased (361 +/- 202 pg/ml); however, only a loose correlation existed between atrial pressures and ANP (right atrial vs ANP, r = 0.03, p = N.S., and pulmonary capillary wedge pressures vs ANP, r = 0.4, p = N.S.). Standing and acute captopril therapy decreased atrial pressures and ANP but did not significantly improve the correlation between atrial pressures and ANP. With chronic captopril therapy, atrial pressures remained decreased; however, ANP had returned to control values (370 +/- 254 pg/ml vs 361 +/- 202 pg/ml) and failed to decrease with standing (370 +/- 259 pg/ml lying vs 351 +/- 194 pg/ml standing). These changes in ANP (with chronic captopril therapy) persisted even once the hemodynamic effects of captopril had dissipated (3 hours post dose). This study indicates that in patients with severe chronic congestive heart failure (1) only a poor correlation exists between ANP and atrial pressures and (2) with chronic captopril therapy ANP increases to higher levels for a given atrial pressure and fails to decrease with standing despite a decrease in atrial pressures. PMID- 2966548 TI - The functional anatomy of the human coronary sinus. AB - The human coronary sinus is an important part of the cardiac venous system that serves as an anatomic landmark as well as a conduit for diagnostic and therapeutic procedures. We studied 50 human coronary sinuses of hearts of normal and increased cardiac weight in order to ascertain various functional anatomic features of the coronary sinus and differences between hearts of normal and increased weight. In the hearts of normal weight (195 to 300 gm) the Thebesian valve covered the ostium of the coronary sinus an average of 41%, with complete coverage in 20%. The valve of Vieussens covered the origin of the coronary sinus an average of 59%. The average volume of the coronary sinus was 1.26 +/- 0.45 cc. In hearts of increased weight (365 to 675 gm), the Thebesian valve covered the ostium an average of 26% and the valve of Vieussens covered the origin of the coronary sinus an average of 56.5%. The average volume of the coronary sinus was increased to 1.76 +/- 0.73 cc (p less than 0.005). Thus knowledge of these functional anatomic features and the differences in hearts of increased cardiac weight allows for better utilization of the human coronary sinus for diagnostic and therapeutic purposes. PMID- 2966549 TI - Acute myocardial infarction after normal maximal exercise test and reperfusion by emergency coronary angioplasty. PMID- 2966550 TI - Changes in coronary blood flow and myocardial metabolism during aortic balloon valvuloplasty. AB - The effects of balloon inflation on myocardial perfusion and metabolism were studied during aortic valvuloplasty in 17 patients with aortic stenosis, including 6 with associated coronary artery disease. Coronary sinus flow and blood samples were obtained before and during the first inflation, and 5 to 10 minutes after the last inflation. During inflation, coronary blood flow decreased (272 +/- 111 standard deviation to 166 +/- 92 ml/min; p less than 0.05), myocardial oxygen uptake fell and transcardiac lactate handling shifted from extraction to production (35 +/- 54 to -41 +/- 48 mumol/min; p less than 0.01). At the end of the procedure, aortic valve area had increased from 0.51 +/- 0.22 to 0.81 +/- 0.48 cm2 (p less than 0.002). Coronary sinus flow increased slightly above control values (+6%; difference not significant) and myocardial oxygen and lactate uptakes were back to control values. However, myocardial alanine production had increased from -3.6 to -6.6 mumol/min (p less than 0.05) and glutamine production was reduced or replaced by extraction (-3.3 +/- 2.1 to 3.5 +/- 3.8 mumol/min; p less than 0.05). Recovery of coronary flow, oxygen and lactate uptakes was not significantly different in patients with or without coronary artery disease, although the former patients tended to have less glutamine extraction and less improvement in their ejection fraction at the end of the procedure. Thus, aortic balloon valvuloplasty produces brief episodes of low-flow ischemia. Recovery of oxidative metabolism is almost immediate after deflation and no detrimental effect seems to persist at the end of the procedure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966551 TI - Electrocardiographic detection of left ventricular hypertrophy in the presence of left anterior fascicular block. AB - The hitherto available electrocardiographic indexes for the detection of left ventricular hypertrophy in the presence of left anterior fascicular block do not provide a reliable diagnosis. Consequently, a new index based on the behavior of the QRS complex in left anterior fascicular block in the frontal and horizontal plane was constructed and its value assessed by echocardiographic measurements. The new index SIII + (R + S) maximal precordial greater than or equal to 30 mm was applied to the electrocardiograms of 50 patients without myocardial infarction and without right bundle branch block, showing a specificity of 87%, a sensitivity of 96%, a positive predictive value of 89% and a negative predictive value of 95%. Echocardiographic measurements were used as reference. Compared with the electrocardiographic indexes used so far (which were also applied to the 50 electrocardiograms), the new index showed a comparable high specificity and a distinctly superior sensitivity. The apparent paradox--why the electrocardiographic diagnosis of left ventricular hypertrophy is easier in the presence rather than in the absence of left anterior fascicular block--is discussed. PMID- 2966552 TI - Acute hemodynamic effects of intravenous isradipine. AB - The acute hemodynamic effects of isradipine, a new dihydropyridine calcium antagonist, were evaluated in 16 men referred for elective cardiac catheterization. Low-dose (0.007 mg/kg, n = 8) and high-dose (0.015 mg/kg, n = 8) isradipine was administered intravenously over 10 minutes and the hemodynamic alterations assessed 10 minutes after completion of infusion. Low-dose isradipine caused increases in heart rate (68 +/- 9 to 79 +/- 12 beats/min, p less than 0.001) (mean +/- standard deviation), cardiac index (3.0 +/- 0.7 to 4.1 +/- 0.9 liter/min/m2, p less than 0.001) and coronary sinus blood flow (114 +/- 27 to 162 +/- 74 ml/min, p less than 0.01), and significant decreases in mean aortic pressure (104 +/- 17 to 92 +/- 10 mm Hg, p less than 0.01), systemic and coronary vascular resistance. High-dose isradipine caused similar effects: the heart rate increased (72 +/- 6 to 84 +/- 14 beats/min, p less than 0.005), as did the cardiac index (3.0 +/- 0.6 to 4.6 +/- 0.9 liter/min/m2, p less than 0.001) and coronary sinus blood flow (122 +/- 48 to 166 +/- 47 ml/min, p less than 0.025). In addition, there were increases in the stroke volume index (43 +/- 10 to 55 +/- 8 ml/m2, p less than 0.001) and left ventricular stroke work index (69 +/- 12 to 79 +/- 12 g-m/m2, p = 0.05) after the high-dose infusion. Vascular resistance declined significantly in the systemic, pulmonary and coronary beds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966553 TI - Heart rate response to intracoronary contrast media injection before and after percutaneous transluminal coronary angioplasty. PMID- 2966554 TI - Usefulness of echocardiographic left ventricular hypertrophy in predicting new coronary events and atherothrombotic brain infarction in patients over 62 years of age. PMID- 2966555 TI - Correlates of supranormal (ejection fraction greater than or equal to 85%) left ventricular performance. PMID- 2966556 TI - Results of percutaneous transluminal coronary angioplasty for unstable angina pectoris in patients 70 years of age and older. AB - Between November 1980 and November 1985, 54 patients ages greater than or equal to 70 years underwent percutaneous transluminal coronary angioplasty for unstable angina, defined as recent-onset (less than 1 month) angina, new onset of rest angina (greater than or equal to 2 episodes) or accelerating class III or IV angina. In these 20 men and 34 women, disease was 1-vessel in 34 (63%) and multivessel in 20 (37%). The mean (+/- standard deviation) ejection fraction was 0.62 +/- 0.12. Angioplasty was successful in 43 patients (80%). In the 11 unsuccessful cases, emergency coronary artery bypass grafting for acute occlusion was performed in 3 and elective coronary artery bypass surgery in 8. There were no deaths. Two patients (4%) sustained Q-wave myocardial infarctions. The mean duration of follow-up for the total group was 37 months (6 to 73 months). Of the 43 patients with successful dilation, 4 died, 1 had an non-Q-wave myocardial infarction and 8 had symptomatic restenosis (4 underwent successful repeat angioplasty, 1 had repeat percutaneous transluminal coronary angioplasty and then bypass surgery, 1 had repeat bypass surgery alone and 2 had medical therapy). At last follow-up, 3 patients had stable class III or IV angina and 31 patients (72%) were angina-free. PMID- 2966557 TI - A symposium: Interventional Cardiology--1987. June 8-9, 1987, Sonoma, California. Proceedings. PMID- 2966558 TI - Interventional cardiology--an overview. PMID- 2966559 TI - Interventional catheterization in pediatric congenital and acquired heart disease. AB - The present status of catheter-directed therapy in pediatric congenital and acquired heart disease is reviewed. Balloon pulmonary valvotomy, established as the procedure of choice for children and adults with isolated pulmonary stenosis, has now been extended to neonates with critical pulmonary stenosis. Balloon mitral and aortic valvotomy are being performed for congenital and acquired stenoses, although indications and optimal techniques remain to be established. Balloon angioplasty of coarctation effectively relieves obstruction in native and post-operative aortic arch stenoses, but remains controversial due to late aneurysm formation. Balloon angioplasty of branch pulmonary artery hypoplasia and stenosis, despite a relatively low success rate and significant complications, remains the procedure of choice due to lack of better therapeutic options. Transcatheter closure of congenital and acquired vessels and defects is being performed with increasing frequency. Embolization of aortopulmonary collaterals and shunts using Gianturco coils is safe and effective in selected patients. Although Rash-kind umbrellas are most commonly used for closure of a patent ductus arteriosus, their use, especially in closing intracardiac defects, remains investigational. PMID- 2966560 TI - Percutaneous transluminal angioplasty in patients with prior myocardial revascularization surgery. AB - Direct myocardial revascularization surgery using either the saphenous vein or internal mammary artery has become the definitive surgical treatment for coronary artery occlusive disease. Certain patients who have undergone these procedures, however, have recurrent myocardial ischemia due to progression of disease in unbypassed vessels, to obstruction in the arteries distal to the insertion of the bypass conduit, or to disease of the conduit itself. Balloon angioplasty may be used to relieve myocardial ischemia in these situations; however, initial studies suggested a low primary success rate coupled with excessive mortality and morbidity. Improvements in patient selection, equipment and technical expertise now allow angioplasty to be performed in this patient population with results comparable to that in the general coronary angioplasty population. Of the 3,016 angioplasty procedures performed between September 1980 and June 1987, 236 patients had previously undergone revascularization surgery. The primary success rate was 93% (390 of 419 stenoses successfully dilated). Overall, clinical restenosis was observed in 39%, including a 43% restenosis rate in patients undergoing only saphenous vein graft angioplasty. This did not differ appreciably from the restenosis rate in postbypass patients undergoing angioplasty of only native vessels (37%) or internal mammary arteries (42%). Emergency revascularization surgery was required in 7 of 236 patients (3%), each of whom had myocardial infarction. One of 236 patients (0.4%) died. Thus, angioplasty may be used to relieve recurrent myocardial ischemia in patients with prior direct myocardial revascularization procedures with a high initial success rate and acceptable risk. Early (less than 6 months) restenosis is not infrequent and remains the largest obstacle to a satisfactory clinical outcome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966562 TI - Angioplasty techniques for stenoses involving coronary artery bifurcations. AB - Branch occlusion during coronary angioplasty is an infrequent but potentially serious complication. The overall incidence of branch occlusion during dilatation of a primary vessel is 5%. Branch vessels most jeopardized by dilatation generally have a complex plaque that not only involves the target vessel but also extends into the origin of the branch vessel. Branches free of pathology at their origin generally have an exceedingly low incidence of occlusion during adjacent balloon dilatation. Side branches at risk for occlusion should be "protected" if the branch vessel is of an important size that could be dilated with a conventional dilatation catheter. The advent of lower profile dilatation catheters and guidewires has provided an opportunity to introduce several pieces of dilatation hardware into the coronary system through a single guiding catheter. Several techniques are described for both "protecting" and dilating side branches, either simultaneously or secondarily, after balloon dilatation of a primary vessel. PMID- 2966561 TI - Initial success and long-term follow-up of percutaneous transluminal coronary angioplasty in chronic total occlusions versus conventional stenoses. AB - Coronary angioplasty was attempted in 1,074 consecutive patients, including 169 patients with total (100%) occlusion (group 1), 102 patients with functional total (99%) occlusion (group 2) and 711 patients with conventional (70 to 95%) stenoses (group 3). After exclusion of 92 patients with acute myocardial infarction, the mean age of the patients was 57 +/- 12 years, including 727 men (74%) and 255 women (26%). Although there were no differences between groups with respect to anginal symptoms or extent of coronary artery disease, the primary success rate (by lesion) varied according to lesion severity, and was 63%, 78% and 90% for groups 1, 2 and 3, respectively (p less than 0.001). Only 3 patients (1%) with chronic coronary artery occlusion (groups 1 and 2) required emergency surgery because of side-branch occlusion in 2 patients and guidewire fracture in 1 patient, whereas emergency surgery was required in 14 patients (2%) in group 3. Long-term follow-up averaging 19 +/- 11 months was available for 95% of patients. Although the combined likelihood of death or nonfatal myocardial infarction was below 6% for each group at 2 years (difference not significant), the likelihood of death, nonfatal infarction, coronary bypass surgery or repeat percutaneous transluminal coronary angioplasty was higher (41%) in group 1 than in groups 2 (28%) or 3 (28%) (p less than 0.001). Thus, coronary angioplasty can be performed safely and effectively in patients with chronic total occlusion, although neither the primary success rate nor the long-term follow-up are as favorable as in patients with conventional stenoses. PMID- 2966563 TI - Use of percutaneous transluminal coronary angioplasty: results of a current survey. AB - To evaluate the current use of percutaneous transluminal coronary angioplasty (PTCA) in the treatment of coronary artery disease, 50 operators experienced in angioplasty (mean experience 949 +/- 706 cases) were surveyed. Although the use of angioplasty is still skewed toward less extensive disease (80% of 1-, 64% of 2 and 39% of 3-vessel disease), it is now estimated that 39 +/- 16% of patients undergoing diagnostic angiography are ultimately referred for PTCA, whereas only 28 +/- 13% are referred to bypass surgery. PTCA thus accounts for 58% of revascularization. Although approximately half of current PTCA patients are clinical candidates for bypass surgery, much of the growth of angioplasty has consisted of treating patients who are either too high a risk for bypass, or who have symptoms with medical therapy that are too mild to warrant surgery. Whereas elective procedures for stable angina (60 +/- 18%) are still the predominant indication for PTCA, a substantial fraction of procedures are performed for unstable angina (27 +/- 10%) or for acute myocardial infarction (13 +/- 15%). Despite a 91% current success rate, 9% of cases are unsuccessful, including 3% in which emergency bypass surgery is required. The most common reasons for unsuccessful procedures are failure to cross the lesion with a wire or balloon (52%), failure to dilate the lesion (4%), and abrupt closure of the dilated segment (44%) due predominantly to extensive local dissection. These data, based on a pooled experience of some 46,000 PTCA procedures, offer a unique glimpse into the current expanded application of this technique. PMID- 2966564 TI - "High-risk" percutaneous transluminal coronary angioplasty. AB - Of 6,500 percutaneous transluminal coronary angioplasty procedures performed between June 1980 and June 1987, 3,501 (1,604 single lesion and 1,897 multiple lesion) were performed in "low-risk" patients with a procedure-related mortality of 0.2 to 0.3%. In comparison, several clinical variables were identified that increased procedural risk by up to 50-fold. These factors include left main dilatation (n = 103, mortality 3.9%), left main equivalent dilatation (n = 77, mortality 2.6%), ejection fraction less than or equal to 40% (n = 664, mortality 2.7%), age greater than or equal to 70 years (n = 1,038, mortality 1.4%), dilatation of all 3 vessels (n = 305, mortality 1.3%), combined diagnostic catheterization and angioplasty for unstable angina (n = 193, mortality 1.5%), and percutaneous transluminal coronary angioplasty for acute myocardial infarction (n = 446, mortality 8.5%). Important considerations in the selection and management of these high-risk patients are discussed. PMID- 2966565 TI - Impact of different reperfusion modalities on ventricular function after acute myocardial infarction. AB - Single-plane contrast ventriculography was performed on admission and before hospital discharge in more than 200 patients with acute myocardial infarction participating in a series of prospective clinical trials including intracoronary streptokinase, percutaneous transluminal coronary angioplasty (PTCA), intravenous tissue plasminogen activator (rt-PA) and thrombolysis (intravenous rt-PA or streptokinase) followed by PTCA. Both global ejection fraction (EF) and regional wall motion of the infarct zone were measured to assess serial changes. Patients treated with intracoronary streptokinase 3.6 +/- 1.8 hours after symptom onset had no increase in EF (mean change 1 +/- 6%, difference not significant [NS]), but patients treated with primary PTCA at 3.0 +/- 1.2 hours did (mean improvement 8 +/- 7%, p less than 0.001). Patients treated with sequential intravenous streptokinase and PTCA 2.6 +/- 1.3 hours after symptom onset showed similar improvement in EF (mean change 6 +/- 12%, p less than 0.002). Patients treated with rt-PA had no change in EF whether treated with rt-PA alone or rt-PA followed by immediate angioplasty (mean change -2 +/- 8% and 0.5 +/- 8%, p = NS, respectively). When angioplasty was used in patients with persistent occlusion after thrombolytic therapy, EF improved in those who had received intravenous streptokinase (mean change 10 +/- 7%, p less than 0.002), but not those who had received rt-PA (+0.5%, p = NS). However, infarct zone regional wall motion improved in patients treated with intracoronary streptokinase (+0.59 +/- 0.79 standard deviation/chord, p less than 0.05), primary PTCA (+1.32 +/- 1.32, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966566 TI - Predictors, therapeutic options and long-term outcome of abrupt reclosure. AB - To ascertain the clinical and procedural factors that relate to the occurrence and management of abrupt reclosure, data on 1,160 consecutive patients who underwent coronary angioplasty between December 1981 and December 1986 were analyzed. Abrupt reclosure occurred in 54 patients (4.7%), developing during the dilatation procedure in 43 patients (80%) and delayed by 11 +/- 2 hours after the procedure in 11 patients (20%). Patients were divided into 3 groups based on subsequent management: (1) 22 patients (40%) in whom the involved vessel was reopened by additional dilatation, (2) 14 patients (26%) in whom the involved vessel was not reopened but the patient was treated medically, and (3) 18 patients (33%) in whom abrupt reclosure was managed by emergency bypass surgery, yielding an overall emergency surgery rate of 1.6%. Patients whose abrupt reclosure was reopened were more likely to have luminal thrombus (54 vs 28%) and less likely to have extensive local dissection after dilatation (14 vs 61%; p less than 0.025) than patients undergoing emergency surgery. Patients treated medically despite continued vessel occlusion were more likely to have collateral blood flow to the occluded zone (54 vs 11%, p less than 0.01) than patients undergoing surgery. While 10 of 18 patients (56%) undergoing emergency surgery sustained a myocardial infarction, all patient subgroups appeared to do well during long-term follow-up. PMID- 2966567 TI - Restenosis after percutaneous transluminal coronary angioplasty. AB - Restenosis after successful percutaneous transluminal coronary angioplasty (PTCA) occurs 4 to 6 months after the procedure in 25 to 30% of the patients. Although PTCA has become far more effective with improved primary angiographic success rates and decreased complication rates, restenosis rates have not changed since the initial experience. Recurrent arterial stenoses appear to be due to fibrocellular proliferation at the site of the initial PTCA. This proliferative response is probably due to platelet adhesion and subsequent activation of the usual tissue injury responses. Fortunately, restenosis seems to be confined to the period soon after the initial PTCA since the long-term, 3- to 8-year studies demonstrate that restenosis occurs infrequently after that. There are certain predisposing characteristics of patients for restenosis: men with a short duration of symptoms with disease of the proximal left anterior descending arteries who are diabetic and continue to smoke cigarettes after PTCA. Inadequate dilatation of the arteries by PTCA and procedures that result in smooth dilatations without any evidence of dissection are associated with increased risk of restenosis. However, most of these patient and procedural characteristics are not controllable. Studies in which procedural and postprocedural variables have been manipulated have been disappointing. Currently, no alterations in techniques or pharmacologic management have proved effective in decreasing the incidence of restenosis. PMID- 2966568 TI - Additional improvement of stenosis geometry in human coronary arteries by stenting after balloon dilatation. AB - The purpose of this study was to assess the early changes in stenosis geometry after insertion of intravascular stents in human coronary arteries. Morphologic changes were evaluated by quantitative coronary angiography (using automated edge detection) and by calculation of the theoretical pressure decrease across the dilated and stented stenosis from the Poiseuille and turbulent resistances assuming a coronary blood flow of either 1 or 3 ml/s. Twenty-six patients were studied before and after angioplasty, as well as immediately after stent implantation. The stented coronary artery was the left anterior descending artery in 19 cases, the circumflex artery in 2 cases, the right coronary artery in 2 cases and a coronary artery bypass vein graft in 3 cases. After stent implantation, an additional increase in minimal luminal cross-sectional area of the dilated vessel was observed, suggesting that the self-expanding stainless steel endoprosthesis used in this study has a dilating function in addition to its stenting role. PMID- 2966569 TI - Perfusion balloon catheter. AB - An autoperfusion balloon catheter was developed to allow passive myocardial perfusion during balloon inflation, through a central lumen and multiple side holes in the shaft proximal and distal to the balloon. This report reviews preliminary experimental animal data and initial human clinical experience with this device. In our first study with this device, the duration of inflation in dogs was compared with the maximal duration of inflation using a standard angioplasty catheter. Coronary arteriography was performed to demonstrate distal coronary blood flow through the perfusion balloon catheter. Electrocardiographic recordings and repeated left ventriculograms were performed to detect evidence of ischemia during standard and perfusion and balloon catheter inflations. The average inflation time was 3 +/- 1 minute for the standard catheter and 37 +/- 10 minutes for the perfusion catheter. Each dog had evidence of severe myocardial ischemia during standard inflation, yet none of the animals had ST-segment elevation, ventricular arrhythmia or wall motion abnormality during dilatation with the perfusion catheter. In a second experiment, the effect of prolonged balloon inflations (30 minutes) on intimal hyperplasia was evaluated in the rabbit model. Results of this study showed reduction of intimal and medial hyperplasia after 4 weeks in iliac arteries in rabbits treated with prolonged inflations compared with the contralateral vessel in rabbits treated with standard angioplasty. Initial clinical results from patients treated with this new catheter are presented. The availability of an effective autoperfusion catheter should allow for testing the hypothesis that prolonged inflations could alter the acute angioplasty success rate and long-term restenosis rate. PMID- 2966570 TI - Role of laser and thermal ablation devices in the treatment of vascular diseases. AB - Since the first coronary angioplasty in 1977, both the number and complexity of interventional procedures have grown dramatically. Continuous-wave and pulsed lasers may further extend the capabilities of balloon angioplasty. Fiberoptic catheters may be used to transmit continuous-wave laser energy to ablate plaque via thermal mechanisms. Pulsed laser systems (such as the excimer) are technologically more complex than the continuous-wave systems, but may prove superior in small vessels given their ability to ablate plaque with minimal associated effects. On the other hand, modifications of the fiber-optic tip, such as the placement of a metal cap, have yielded even better results than current bare fiber systems. Such laser thermal techniques have proved a useful adjunct to balloon dilatation in peripheral vessels, but further research is necessary to determine their effect on coronary arteries. New, nonlaser technologies, however, may provide simpler power sources for thermal angioplasty. Although balloon angioplasty remains the cornerstone of interventional vascular therapy, new technologies should help to further expand the indications for nonsurgical interventions. PMID- 2966571 TI - Advances in balloon technology and reperfusion devices for peripheral circulation. AB - Major improvements in balloon and wire technology have been occurring for several years and, consequently, the approach to conventional angioplasty has been simplified. Results in isolated stenotic segments in renal circulation and within the iliac and superficial femoral vessels have generally been 95% technically successful, with 1-year patency rates of 80% in the superficial and renal sites and 90% in the iliac vessels. The procedure has been extended to include more distal vessels including the popliteal and tibial circulation as well as more complex lesions involving the mesenteric circulation and pre-Leriche syndromes with stenotic disease involving the infrarenal segment of the abdominal aorta. Successful results have also been achieved in stenotic atherosclerotic disease involving the subclavian artery and the associated subclavian steal syndromes. Preliminary results are also being reported in a few selected series of vertebral and carotid artery angioplasty. Totally occluded segments in the iliac and peripheral circulation had previously been ammenable only to surgical bypass procedures. A series of new recanalization devices are under investigational evaluation for reperfusing these totally obstructed segments. Although technical recanalization in segments as long as 20 cm can be accomplished in approximately 75% of these patients, long-term patency is totally dependent on popliteal-tibial outflow. For these reasons, diffuse disease and totally obstructive disease in the superficial femoral vessels can have recurrence rates as high as 45%, and reestablishing tibial outflow is equally as important as successfully recanalizing the totally obstructed superficial femoral vessels. PMID- 2966572 TI - Role of percutaneous transluminal coronary angioplasty in multivessel disease. AB - Although percutaneous transluminal coronary angioplasty (PTCA) is being widely performed in patients with multivessel disease, the eventual role it will play will depend on several factors, including the immediate and long-term results, procedural risks and restenosis. An important consideration is that of completeness of revascularization. This is based on cardiac surgical experience, which has documented that if revascularization is complete, the clinical outcome will be improved. The importance of this concept has been borne out in practice. Although complete revascularization is ideal, it cannot be achieved in a substantial number of patients with multivessel disease because of the presence of old total occlusion that cannot be dilated, diffuse and distal disease or a planned dilation strategy. However, many patients with successful dilation but incomplete revascularization do well. In these patients, attempts are made to identify and then dilate a "culprit" lesion. Dilation of these most physiologically important stenoses often results in an excellent short-term outcome. Currently, 2 studies have been initiated to compare the role of PTCA with that of coronary artery bypass grafting for the treatment of patients with multivessel disease. For these studies, initial success and long-term outcome in terms of morbidity and mortality as well as cost considerations will be assessed. The results of these studies will help to put into perspective the complementary roles of PTCA and coronary artery bypass grafting. PMID- 2966575 TI - Cruveilhier-Baumgarten syndrome. PMID- 2966574 TI - Plasma concentrations of immunoreactive atrial natriuretic peptide in hospitalized cirrhotic and noncirrhotic patients: evidence for a role of deficient atrial natriuretic peptide in pathogenesis of cirrhotic ascites. AB - Atrial natriuretic peptide(s) (ANP), are thought to be released from the cardiac atria in response to distension. If decreased effective circulating blood volume is important in pathogenesis of ascites, plasma ANP levels would be expected to be decreased in ascitic subjects because of decreased atrial distension. To test this hypothesis, we measured plasma ANP by competitive radioimmunoassay in three groups of fasted, supine hospitalized subjects: nine noncirrhotic control subjects, 12 cirrhotics without ascites, and 17 cirrhotics with moderate to marked ascites. Immunoreactive plasma ANP concentrations were 195 +/- 41, 171 +/- 31, and 137 +/- 34 pg/ml (m +/- SD), respectively, in the three groups. The mean concentration in the group with cirrhosis and ascites was significantly (p less than or equal to 0.01) les than those of the other two groups, which did not differ from one another. These results support the concept that decreased effective circulating volume plays a role in pathogenesis of cirrhotic ascites, and that a relative deficiency of ANP plays a role in the sodium retention of decompensated cirrhosis. PMID- 2966573 TI - Transluminal atherectomy for occlusive peripheral vascular disease. AB - Sixty-one patients with occlusive peripheral vascular disease were treated with transluminal atherectomy, a catheter-mediated technique for removal of atheroma. The technique was performed using 7Fr, 9Fr or 11Fr atherectomy catheters. Mean percent diameter stenosis was reduced from 71 to 23%, by removal of 831 atheromatous specimens in 949 passes of the cutting element through 136 stenoses in 61 patients. All specimens removed were sent for histopathologic examination to determine the components of the atheroma removed, which differed for specimens removed from original vs restenotic lesions. Percent stenosis was reduced to less than 45% in 118 of 136 stenoses (87%). Complications included 1 thrombus, which resolved after intraarterial infusion of streptokinase and 1 probable distal embolization without sequelae. Three angiographic dissections occurred without impairment of blood flow. There were no instances of acute occlusion, vascular spasm or vessel perforation. Six-month follow-up angiography was performed showing that patients who had a residual stenosis less than 30% after initial atherectomy had a lower restenosis rate (18%) than patients with initial residual stenoses greater than 30% (52%); this result demonstrated the importance of performing more complete atherectomy. Transluminal atherectomy appears to be an effective, predictable and safe method for removing occlusive atheromatous deposits from peripheral arteries. PMID- 2966576 TI - Immunological profile of chest x-ray-negative, asymptomatic asbestos workers. AB - Several immunologic parameters, both humoral and cellular, were studied in the serum and peripheral blood lymphocytes derived from chest x-ray-negative, asymptomatic asbestos workers. All humoral and cellular parameters were intact, except the con-A-induced T cell suppressor activity and T cell division in autologous mixed lymphocyte reaction, which were significantly elevated in the asbestos plant workers. The significance of these increased T cell activities in asbestos exposed people is not clear, and further clinical and immunological follow-up is warranted. PMID- 2966577 TI - Assuming the risks: occupational disease in the years before workers' compensation. AB - In the late 19th and early 20th centuries, new knowledge of work-related illness became part of discourses in several institutional spheres on the relationship between aspects of the economy and workers' health. Appeals courts and state legislatures invoked this knowledge in their deliberations on legislation to ease the coercive aspects of the employment relationship. Insurers used the knowledge to help determine what types of coverage would be available to different occupational groups. In the courts, a narrow compensation remedy evolved for illness caused by employers' failure to warn of latent risks. This small, legal zone of protection of workers' health was separated out from the massive amount of uncompensated, preventable work-related illness. PMID- 2966578 TI - Something better for occupational disease victims. PMID- 2966579 TI - Abnormalities in CD4+ T-lymphocyte subsets in inflammatory rheumatic diseases. AB - The monoclonal antibodies anti-2H4 and anti-4B4 identify the suppressor-inducer (CD4+2H4+) and helper-inducer (CD4+4B4+) subpopulations of CD4 (T4+) lymphocytes, respectively. The cell surface phenotype of peripheral blood lymphocytes and synovial fluid lymphocytes in patients with rheumatoid arthritis and other inflammatory joint diseases was analyzed by use of these and other well characterized anti-T-cell monoclonal antibodies. In the synovial fluid of patients with rheumatoid arthritis, there was a markedly decreased percentage of T4+2H4+ suppressor-inducer cells (3.1 +/- 1 percent) and an increased percentage of T4+4B4+ helper-inducer cells (29.1 +/- 9 percent) as compared with the proportions found in the peripheral blood of normal individuals (T4+2H4+: 19.0 +/ 6 percent, T4+4B4+: 23.0 +/- 7 percent). Moreover, patients with other chronic and acute inflammatory joint diseases exhibited highly similar synovial T-cell findings to those of the patients with rheumatoid arthritis (T4+2H4+: 4.2 +/- 3 percent, T4+4B4+: 33.1 +/- 9 percent). In contrast, there were no significant differences between the normal control subjects and patients with rheumatoid arthritis in the percentage of T4+2H4+ cells in peripheral blood lymphocytes, nor were there significant differences between normal control subjects, patients with rheumatoid arthritis, and patients with other joint diseases (osteoarthritis, gout, B27+ spondyloarthropathy, and psoriatic arthritis) in the number of T4+4B4+ cells or in the T4/T8 ratio of peripheral blood lymphocytes. However, very low numbers of T4+2H4+ (suppressor-inducer) peripheral blood lymphocytes were seen in a subgroup of patients, including five of seven with Reiter's syndrome and several patients with systemic rheumatic disease syndromes. In addition, although the percentage of T4+2H4+ cells in peripheral blood lymphocytes of patients with osteoarthritis (13.7 +/- 7 percent) and gout (14.3 +/- 7 percent) was decreased compared with that of normal controls (19.0 +/- 6 percent) (osteoarthritis versus normal controls p less than 0.025), this difference appeared to reflect alterations due to age rather than disease. Consistent with the phenotypic changes observed, synovial T cells were also functionally defective, since autologous mixed lymphocyte reaction-activated T4 cells from the synovial fluid of patients with rheumatoid arthritis failed to exhibit suppressor-inducer activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966580 TI - Role of atriopeptin in blood pressure regulation. PMID- 2966581 TI - The atrial natriuretic factor in hypertension. AB - Plasma immunoreactive atrial natriuretic factor (IR-ANF) concentration measured by radioimmunoassay after extraction on Sep-Pak cartridges was studied in 64 control normotensive subjects, 25 patients with labile essential hypertension, 67 patients with mild essential hypertension (diastolic pressure between 90 and 105 mm Hg and no left ventricular hypertrophy) and 9 patients with moderate to severe essential hypertension (diastolic pressures between 105 and 120 mm Hg). An additional group of 16 patients under medication but without effective control of their blood pressure and with diastolic pressure above 110 mm Hg also was studied. Results show that plasma IR-ANF concentrations are within normal range in patients with labile, mild, and moderate hypertension. In view of the reported increased right and left atrial pressures and distension in patients with mild and moderate hypertension, these findings strongly suggest a state of hyporesponsiveness of the atria to release ANF. PMID- 2966582 TI - On-the-job back exercises. PMID- 2966583 TI - Congenital defects of abdominal wall. PMID- 2966584 TI - Selective inhibition of c-myc expression by the ribonucleic acid synthesis inhibitor mithramycin. AB - Expression of the c-myc proto-oncogene has been shown to correlate with the rate of cellular proliferation and malignant transformation in a number of cell types. JEG-3 choriocarcinoma cells demonstrate c-myc transcript levels that are greater than those of nonmalignant trophoblastic tissue at any stage of gestation. Southern blot analysis documents c-myc gene amplification in JEG-3 cells, with a gene copy number of approximately 20. The methylation pattern and genomic structure of the amplified c-myc oncogene in JEG-3 choriocarcinoma cells are identical to those of normal placenta. Treatment of JEG-3 cells with mithramycin, a ribonucleic acid synthesis inhibitor, results in a dramatic decrease in c-myc expression relative to that of the c-Ha-Ras gene. The apparent selectivity of mithramycin for c-myc expression represents the only example, to date, of the selective pharmacologic modulation of oncogene expression. PMID- 2966585 TI - Pulsed Doppler flow-velocity waveforms before and after intrauterine intravascular transfusion for severe erythroblastosis fetalis. AB - Pulsed Doppler studies of the fetal and maternal circulations were carried out before and after 64 intrauterine transfusions performed on 24 fetuses. A model was derived for the prediction of hematocrit before the first transfusion: Hematocrit = 7.778 - (0.088 x peak velocity in descending aorta) + (0.968 x gestational age [weeks]) - (10.911 if hydrops present) (r = 0.876, p less than 0.0001). An alternative formula, excluding hydrops, was slightly less predictive: Hematocrit = 45.312 - (56.261 x umbilical cord Pourcelot index) - (0.128 x peak velocity in descending aorta) + (1.042 x gestational age) r = 0.822, p less than 0.001). Neither model was accurate in the prediction of hematocrit before second or subsequent transfusions. A third model was derived from second-transfusion data: Hematocrit = 40.524 - (0.045 x peak velocity in descending aorta) - (10.693 x pulsatility index of maternal uterine artery) (r = 0.81, p less than 0.003). However, this model was unable to predict hematocrit before third or later transfusions. No changes in Doppler parameters before and after transfusion were found. We conclude that pulsed Doppler ultrasound may be helpful in the evaluation of isoimmunized pregnancies, in differentiating anemic from normal fetuses. It does not appear to be useful in determining the timing of later transfusions. The lack of change before and after transfusions suggests that these vessels will not provide significant information concerning the effect of rapid volume and hematocrit changes in the fetus. PMID- 2966586 TI - Steroid receptors and clinical outcome in patients with adenocarcinoma of the endometrium. AB - Progesterone receptor content was measured in tissue samples from 175 patients with endometrial adenocarcinoma by use of the dextran-charcoal method. The estradiol receptor content was determined in 138 of these samples. Ninety-two tumors (52.6%) tested positive for progesterone receptors (greater than 50 fmol/mg cytosol protein) and 111 (80.4%) tested positive for estradiol receptors (greater than 6 fmol/mg). Median follow-up was 27.3 months (range 1 to 152 months). Progesterone receptor status correlated significantly with grade, histology, adnexal spread, age, and recurrence rate in stage I cancer. There was no correlation between progesterone receptor status and clinical stage, myometrial invasion, peritoneal cytology, retroperitoneal lymph node involvement, or spread to the cervix. Estradiol receptor status correlated with adnexal spread and recurrence rate. Recurrence in patients with stage I disease was significantly more common if tumors were negative for progesterone receptor (16 of 43, 37.2%) than if they were positive (four of 57, 7%; p less than 0.001). Recurrence was also more common if tumors were negative for estradiol receptor (seven of 17, 41.2%) than if they were positive (eight of 63, 12.7%; p = 0.02). In recurrent or advanced disease, response to progestin was independent of estradiol receptor content, but tumors positive for progesterone receptors responded significantly more often than those lacking progesterone receptors. Overall survival was superior for patients with progesterone receptor-positive tumors (p = 0.001). Although survival in clinical stages I and II was also superior in patients with lesions positive for progesterone receptors (p = 0.13), there was no statistical difference in survival between patients with progesterone receptor-positive or -negative cancers and surgical stages I and II disease (p = 0.12). Estradiol receptor status had no apparent correlation with survival. PMID- 2966587 TI - Use of an agonistic analog of gonadotropin-releasing hormone (nafarelin) to treat leiomyomas: assessment by magnetic resonance imaging. AB - The purposes of this study were to investigate the effect of a superactive agonistic analog of gonadotropin-releasing hormone, nafarelin, on uterine leiomyomas and to assess the use of magnetic resonance imaging in monitoring uterine and myoma size. Eleven women with uterine leiomyomas were treated with 800 micrograms of nafarelin per day for 6 months. Serum gonadotropin and estradiol concentrations were suppressed during treatment. The mean +/- SEM serum luteinizing hormone level decreased from 11.1 +/- 1.4 to 5.6 +/- 0.42 mlU/ml and follicle-stimulating hormone from 9.5 +/- 0.66 to 7.5 +/- 0.72 mlU/ml by 3 months of treatment (p less than 0.01). The estradiol level decreased from a pretreatment follicular phase mean +/- SEM of 43 +/- 8.3 to 19.8 +/- 3.1 (p less than 0.05) and 14.8 +/- 2.2 pg/ml (p less than 0.01) at 3 and 6 months of treatment, respectively. Mean pretreatment androgen levels (testosterone, androstenedione, and dehydroepiandrosterone sulfate) were low in these women and did not change significantly during treatment. Ten women had magnetic resonance imaging, which provided excellent resolution of individual uterine myomas. As assessed by magnetic resonance imaging, the largest myoma decreased in size in nine of 10 women; the mean decrease was 46% +/- 9%. Uterine volume decreased in all 10 patients; the mean decrease was 57% +/- 7%. In several women myomas reenlarged after discontinuance of nafarelin treatment. Posttreatment myomectomy was carried out in four women; there was minimal blood loss and no surgical complications. These data indicate that suppression of ovarian estrogen production with nafarelin is associated with a decrease in uterine myoma size in many women but that myomas may regrow with reinstitution of ovarian function. Magnetic resonance imaging is an excellent method by which to monitor treatment as changes in the size of the uterus, as well as individual myomas, can be assessed. The optimal use of gonadotropin-releasing hormone analogs may be in perimenopausal women or as presurgical treatment to decrease uterine and myoma size to facilitate myomectomy. PMID- 2966588 TI - Qualitative assessment of maternal uterine and fetal umbilical artery blood flow and resistance in laboring patients by Doppler velocimetry. AB - The purpose of this study is to evaluate the effect of uterine contractions during labor on both the uterine and the umbilical circulations. Twenty-seven patients in active labor were studied by continuous-wave Doppler velocimetry. Umbilical, left uterine, and right uterine arterial waveforms were obtained before, during, and after peak uterine contractions, and the ratio of maximum systolic and minimum diastolic velocities was calculated. Fifteen patients showed absent flow in end diastole on the uterine artery waveform and had significantly higher intra-amniotic pressures (64.5 +/- 3.5 mm Hg) during the peak amplitude of the uterine contraction compared with the 12 patients with maintained end diastolic flow (46.5 +/- 2.6 mm Hg; p less than 0.05). During the peak amplitude of the uterine contractions the 12 patients maintaining end-diastolic flow had significantly higher systolic/diastolic ratios in the uterine artery (6.5 +/- 1.5) compared with either before or after a contraction (2.1 +/- 0.15 and 2.0 +/- 0.2, respectively; p less than 0.05). Also, these 12 patients showed a linear relationship between the systolic/diastolic ratio and the intrauterine pressure. However, no differences were observed in the umbilical artery systolic/diastolic ratios before, during, or after a contraction in the intensity range studied. On the contrary, during contractions an increase in uterine artery resistance occurs with decreased or absent end-diastolic flow, which bears an inverse linear relationship to the intensity of the contraction. This suggests that during uterine contractions the human fetus continues to have uninterrupted fetoplacental blood flow, whereas the degree of interruption or reduction in uteroplacental blood flow is dependent on the intensity of uterine contraction. PMID- 2966589 TI - Indications of a differentiated regulation of sound transmission by the middle ear muscles of the rat. AB - Measurement of the middle ear muscle reflex is an important tool in audiologic examination; however, the precise function of the tensor tympani and stapedius muscle is not fully understood. The function of the middle ear muscles in speech discrimination and noise-induced hearing loss is the main object of our study. Morphologic and enzyme-histochemical properties of the middle ear muscles of the rat indicate a complex, fine-tuned function of the middle ear muscles. Both middle ear muscles are merely composed of relatively small fast-twitch fibers. Almost all fibers possess an enzymatic profile that allows aerobic as well as anaerobic metabolism. The innervation of the muscles is extensive, and motor end plates are well developed. In a parallel study, middle ear muscle contraction (to be analyzed by electromyography) will be correlated with a change of the sound transmission characteristics of the middle ear (measured by the electrocochlear and brainstem auditory evoked responses). Preliminary results of the electrophysiologic measurements (electrocochleogram and brainstem auditory evoked response) are presented. PMID- 2966590 TI - Oxidative potential in developing rat diaphragm, EDL, and soleus muscle fibers. AB - To examine the effect of postnatal development on changes in oxidative potential of fibers of specific types (I, IIa, IIb, and IIc) in the rat diaphragm, determinations of succinate dehydrogenase (SDH) activity were made using microphotometric measures of optical density. Samples of the costal region of the diaphragm were extracted from 56 male Wistar rats ranging in age from 8 to 85 days and subgrouped into seven developmental periods (1, 2, 3, 4, 6, 9, and 12 wk). For type I fibers, increases of 17% (P less than 0.05) in SDH activity occurred during 2nd wk, remained elevated through 4th wk, and increased further (P less than 0.05) to 137% of 1-wk values by the end of 6th wk. No further increases were noted between 6 and 12 wk. A similar maturational trend was evident for type IIa fibers, although SDH activities remained higher throughout development when compared with type I fibers. In contrast, SDH in type IIb fibers, although increasing by 14% during the first two measurement weeks (P less than 0.05), declined from 6 to 9 wk before ultimately reaching a value similar to 3 wk. SDH activity was also assessed in a typical slow- (soleus) and fast-twitch (extensor digitorum longus, EDL) muscle of the hindlimb to contrast their development with that of the diaphragm. Generally, SDH in type I and IIa fibers was approximately 40 and 20% higher, respectively, in the diaphragm than in matched fiber types in the other muscles throughout development (diaphragm greater than EDL greater than soleus).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966591 TI - Transport of L-proline by luminal membrane vesicles from pars recta of rabbit proximal tubule. AB - The mechanism of renal transport of L-proline by luminal membrane vesicles prepared from proximal straight tubules (pars recta) of rabbit kidney was investigated. The following picture emerges from transport studies: an electrogenic and Na+-requiring system confined to this region of nephron exists for transport of L-proline with a high affinity (Km = 0.16 mM) and low capacity (Vmax = 3.5 nmol.mg protein-1.15 S-1). Lowering the pH from 7.5 to 5.5 increased the affinity (Km lowered from 0.16 mM at pH 7.5 to 0.08 mM at pH 5.5) without changing the maximal capacity of this system. Modification of histidyl residues of the intact luminal membrane vesicles by diethyl-pyrocarbonate (DEP) completely abolished the transient renal accumulation of L-proline. Simultaneous presence of Na+ and L-proline (10 mM) protects against DEP inactivation of renal transport of radioactive L-proline. We propose that a histidyl residue may be at or close to the active site of L-proline transporter in vesicles from the pars recta. PMID- 2966592 TI - Hormonal determinants of sodium excretion in rats with experimental high-output heart failure. AB - The present study evaluates the inter-relationship between the alteration in atrial natriuretic factor (ANF) and the renal handling of Na in rats with chronic aortocaval (a-v) fistula, an experimental model of congestive heart failure. Balance studies in these animals showed two distinct patterns of Na excretion: some rats developed progressive Na retention [urinary sodium excretion (UNaV) less than 100 mueq/24 h], whereas others compensated and returned to normal Na balance (UNaV greater than 1,200 mueq/24 h). Base-line plasma ANF levels were equally elevated in Na-retaining and compensated rats with a-v fistula (588 +/- 70 vs. 621 +/- 114 pg/ml, P, NS). However, the response of the two groups to exogenous administration of synthetic rat ANF-(101-126) in incremental doses varied greatly. ANF infusion increased the fractional Na excretion (FENa) in compensated animals from 0.12 +/- 0.03 to 2.6 +/- 0.5%, whereas the rise in FENa in Na-retaining animals was markedly blunted (0.11 +/- 0.06 to 0.89 +/- 0.35%). A similar pattern of ANF action was observed on the glomerular filtration rate and urine flow. The blunted response to ANF in the Na-retaining animals was associated with a marked increase in plasma renin activity (PRA) (35.6 +/- 6.9 vs. 4.5 +/- 0.7 ng ANG I.ml-1.h-1 in sham control rats, P less than 0.05) and plasma aldosterone levels (729.3 +/- 28.2 vs. 42.6 +/- 18.4 ng/dl in sham control rats, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966593 TI - Interrelationships among atrial peptides, renin, and blood volume in pregnant rats. AB - To determine how changes in intravascular volume are sensed by atrial and renal volume receptors during pregnancy and the puerperium, circulating atrial natriuretic peptide (ANP) levels and plasma renin activity (PRA) were measured in conscious chronically catheterized rats on days 9-10, 15-16, 19-20, and 22 of pregnancy, on the first postpartum day, and in nonpregnant controls. Blood volume measured in a separate group of anesthetized rats increased progressively during pregnancy, and circulating ANP levels tended to decline, although not significantly below the nonpregnant value of 132 +/- 9 pg/ml (mean +/- SE). PRA remained similar to the nonpregnant value of 5.5 +/- 0.6 ng angiotensin I.ml-1.h 1 until day 22 of pregnancy, when it rose to 14.1 +/- 1.8 ng angiotensin I.ml-1.h 1 (P less than 0.001 vs. nonpregnant). In pregnant rats, PRA was suppressed after uninephrectomy and chronic administration of deoxycorticosterone and saline, demonstrating the ability of this hormonal system to respond appropriately to further increments in volume induced during pregnancy. On the first postpartum day plasma ANP rose to 268 +/- 26 pg/ml, and PRA fell to 3.6 +/- 0.4 ng angiotensin I.ml-1.h-1 (P less than 0.005 and P less than 0.05 vs. nonpregnant values, respectively). Thus it appears that the increased blood volume in normal pregnancy is not sensed by renal or atrial volume sensors, presumably because it is accommodated by an enlarged maternal vascular compartment. In the puerperium, however, due to the decreased size of the maternal vascular compartment, atrial and renal volume sensors recognize the intravascular volume as expanded.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966594 TI - Monitoring the neuromuscular junction. PMID- 2966595 TI - Radioreceptor assay for atrial natriuretic factor. AB - Interest in accurate measurement of atrial natriuretic factor (ANF) in biological fluids and various tissues has been stimulated by recent data indicating the possible role of ANF in the homeostasis of salt and water. The presence of high affinity binding sites for ANF in rat glomeruli has allowed us to develop a rapid, sensitive, and simple radioreceptor assay (RRA). A saturable high-affinity binding site on the membranes of rat glomeruli has been characterized by a dissociation constant of 33 pM and binding capacity of 396 fmol/mg protein. Rat plasma extracts or atrial homogenates or standards were incubated with radioiodinated ANF and a preparation of rat glomerular membranes. The receptor bound and free radioactivity were separated by filtration on Whatman GF/C paper after 1 h incubation at room temperature. The sensitivity of the RRA was 2.08 fmol. The effective concentration of standard ANF that displaced 50% of labeled receptor-bound ANF (EC50) was 43.3 +/- 2.6 fmol/ml (n = 7). Both intra- and interassay coefficients of variation were smaller than 11%. This RRA assay has been compared with radioimmunoassay (RIA). High correlations for 19 plasma extracts and 34 atrial homogenates (r = 0.973 and r = 0.954, respectively) tested by RRA and RIA were obtained. This good correlation between the two methods suggests that the immunoreactive material found in rat plasma and atrial homogenates also displays biological activity. PMID- 2966596 TI - Kinetics of vanadate dissociation: estimation of the rate by inhibitor inactivation. AB - Vanadate (+5) is a potent inhibitor of a variety of ATPases including dynein ATPase. We describe a method useful for estimating the functional dissociation rate of vanadate from the active site which does not rely on classical physical separation techniques. The method involves spectrophotometrically monitoring the enzymatic activity as the inhibitor dissociates from the enzyme and is inactivated by norepinephrine. Norepinephrine effectively reverses vanadate inhibition by reducing vanadate (+5) to oxovanadium (+4). This reduction by norepinephrine is sufficiently fast for these purposes--addition of vanadate after norepinephrine shows no inhibition of ATPase activity. The mathematical estimation procedure is generally useful for estimation of dissociation rates of other reversible inhibitors which can be quickly inactivated after dissociation from the enzyme. The rate of dissociation of vanadate from dynein with ATP and 2 N3ATP as substrates using this method was estimated to be in the ranges 0.0023 0.0042 and 0.0057-0.0075 s-1, respectively. These rates permit estimation of the rates of vanadate association with dynein by using the reported dissociation constant for vanadate. The results are consistent with the very fast and potent inhibition of dynein ATPase activity observed. PMID- 2966597 TI - Ultrastructural identification of the primitive muscle spindle in the Xenopus laevis larvae. AB - The initial formation of muscle spindles was studied with electron microscopy using the toe muscle of Xenopus laevis. At the larval stage 57 (Nieuwkoop and Faber 1967), muscle spindles were first identified primarily by the presence of sensory endings associated with a thin bundle of myotubes, e.g. intrafusal (IF) myotubes which were partly invested by a single cellular layer. The number of IF myotubes per spindle was 5 to 6; the adult complement. IF- and extrafusal (EF) myotubes were almost identical in their size and structure. A few thinner IF myotubes with scarce myofibrils were also present. The reticular zone had been undeveloped. Sensory endings were smaller in size and in number per spindle than those in the adult, forming irregular beaded chains with occasional tubular expansions. The endings and IF myotubes were rarely in direct contact, being frequently interposed by a satellite cell and its process. Incipient fusimotor endings were widely distributed from the juxta-equatorial to the polar region. Large cored vesicles resembling the neurosecretory vesicles occurred in sensory and motor endings as well as in intramuscular nerve fibers. The vesicles may be involved in the neuronal influence upon the spindle differentiation. The results were compared with the formative process of mammalian spindles. PMID- 2966598 TI - Motor and sensory innervation of muscle spindles in the neonatal rat. AB - Neural and muscular elements of three muscle spindles from the soleus muscles of 4-day-old rats were reconstructed by electron microscopy of skip-serial transverse ultrathin sections. Each spindle contained four encapsulated intrafusal fibers, including a minimum of one bag1, one bag2 and one chain fiber. The fibers were innervated by unmyelinated motor and sensory axons. The primary and secondary afferents approached the spindles as single axons and terminated on the central region of the intrafusal fibers. Single profiles of terminal axons occupied the sites of sensory neuromuscular junctions, similar to adult sensory endings. No morphological features suggested retraction of afferents from 4-day postnatal spindles. Motor axons approached spindles tightly packed in bundles of 5-20 axons and terminated in the juxtaequatorial and polar regions of both bag and chain fibers. Multiple profiles of terminal axons were visible for each intrafusal motor ending. More motor axons innervated 4-day postnatal spindles and a greater number of axon terminals were visible in immature intrafusal motor endings than in adult spindles. The data suggest that postnatal maturation of motor innervation to intrafusal fibers involves the elimination of supernumerary motor nerve inputs. Synapse elimination in the development of the fusimotor system might represent a mechanism whereby individual gamma axons adjust the number of spindles they innervate. PMID- 2966599 TI - The anionic matrix at the rat glomerular endothelial surface. AB - The anionic macromolecules at the glomerular endothelial cell surface are visualized only when stained with cationic stains. We investigated the arrangement and composition of this anionic matrix at the luminal surface. Rat kidneys were perfused with anionic ferritin (pI 4.5), ferritin (pI 7.4), or cationized ferritin (CF, pI 8.3). Anionic ferritin (pI 4.5) did not bind to the capillary wall, ferritin (pI 7.4) bound discontinuously only to the laminae rarae of the basement membrane, but cationized ferritin (CF, pI 8.3) bound as a thick continuous layer to the cell plasmalemma and bound to the anionic matrix in the fenestral spaces. These observations show that an anionic matrix lines the entire capillary lumen surface, fills the fenestrae, and is interposed between the blood and the basement membrane at the fenestrae. The anionic constituents at the capillary luminal surface were identified by in vivo digestion with specific enzymes. Absence of CF binding following digestion with specific enzymes was taken to indicate the presence of the particular glycoprotein known to be susceptible to the enzyme used. Neuraminidase digestion revealed that anionic sites over the surface plasmalemma are mainly from sialoproteins. In contrast, the matrix in fenestral channels contains heparan sulfate, hyaluronic acid, and sialoproteins. Papain digestion showed no glycolipids at the luminal surface. The functions of this continuous anionic layer located at the luminal surface of glomerular capillaries have not yet been established. PMID- 2966600 TI - Continuous infusion epidural analgesia in labor: the effect of adding sufentanil to 0.125% bupivacaine. AB - The purpose of this study was to determine if the addition of sufentanil would improve the quality of analgesia obtained with 0.125% epidural bupivacaine infusions given to women in active labor. Forty healthy laboring women were randomly assigned to two equal groups. Group 1 had epidural analgesia instituted with the bolus injection of 10-15 ml 0.125% bupivacaine containing sufentanil 2 micrograms/ml, followed 30 minutes later by initiation of an epidural infusion of 0.125% bupivacaine containing sufentanil 1 microgram/ml at a rate of 10 ml/hr. Group 2 had epidural analgesia instituted with 10 or 15 ml of 0.25% bupivacaine with an epidural infusion of 0.125% bupivacaine begun 30 minutes later at a rate of 10 ml/hr. Infusion rates were altered as required to maintain an analgesic level to the tenth thoracic dermatome. Analgesia assessed by visual analog pain scores was significantly better in group 1. Significantly fewer epidural injections were required in group 1, and less motor weakness occurred in these patients. The addition of sufentanil to epidural bupivacaine infusions given in labor improves analgesia and reduces "top-up" requirements. PMID- 2966601 TI - Application of tapered teflon dilator to PTCA. AB - A technique using a tapered teflon dilator was applied to percutaneous transluminal coronary angioplasty (PCTA). Seven of 10 cases were successfully dilated by using this technique. These lesions could not be dilated in the usual manner using standard PTCA balloons. This technique converted 7 of 10 failed procedures into technical successes and is the basis of this report. PMID- 2966602 TI - [Atrial natriuretic factor. Physiological properties and physiopathological role]. PMID- 2966603 TI - Regionally selective manifestations of neostriatal aging. AB - Presynaptic and postsynaptic markers of the cholinergic and dopaminergic systems have characteristic topographical distributions within the striatum. Aside from the dopaminergic afferents, several other afferent systems exhibit a heterogeneous distribution in the striatum. The net result is that each part of the striatum receives a specific and unique combination of afferents. Moreover, the intrinsic striatal systems also have unique distributions, so each part of the striatum consists of a unique combination of afferent and intrinsic neurotransmitter systems. In view of these points, one may expect that the striatum is functionally very complex, integrating information from a wide variety of brain areas. One may also assume from these facts that the striatum is a functionally heterogeneous structure. Consistent with that conclusion, behavioral and pharmacological studies show that interruption of neurotransmission in localized regions of the striatum produces very specific behavioral and physiological effects. Age-related neurochemical changes are also confined to specific striatal regions. Which regions are affected will depend on a variety of factors, including the neurochemical parameter studied and the species or strain of animal. However, we still do not know what factors make a particular striatal area vulnerable to the effects of aging or disease. Moreover, a question that remains to be answered is whether the regions that are affected by neurodegenerative diseases are the same ones affected during normal aging. If so, then this may provide a clue as to why neurodegenerative diseases of the basal ganglia increase in frequency with advancing age. Nevertheless, discrete regional neurochemical alterations may underlie specific symptoms of these diseases. Further study of this relationship may provide the basis for treatments that better target the source of the symptoms. Not only would this increase the effectiveness of the treatment, it would help reduce potential side effects. This may be particularly important, for example, with respect to the use of tissue explants in the treatment of diseases of the basal ganglia. PMID- 2966604 TI - Agonist binding to striatal dopamine receptors in aging. The ternary complex of receptor and guanine nucleotide binding regulatory protein. PMID- 2966605 TI - In vivo measurement of dopamine receptors in human brain by positron emission tomography. Age and sex differences. PMID- 2966606 TI - Toxicity of trimetrexate on hemopoietic progenitor cells in normal mice. AB - Single increasing doses of methotrexate (MTX) and trimetrexate (TMQ) were administered to normal mice. Survival of hemopoietic progenitor cells assayed as CFU-S and GM-CFC was determined 24 hr after drug injection. The survival of each population in TMQ-treated animals was not statistically different from that observed in mice treated with MTX. No difference was observed in time-survival curves of hemopoietic progenitor cells comparing TMQ to MTX. TMQ toxicity at the hematological level thus seems comparable to that of MTX. PMID- 2966608 TI - In vitro antibacterial activity of trospectomycin (U-63366F), a novel spectinomycin analog. AB - Trospectomycin (U-63366F) is a novel spectinomycin analog with broad-spectrum antibacterial activity. The in vitro activity of this analog was compared with that of spectinomycin and other reference antibiotics against 411 clinical isolates of aerobic and anaerobic bacteria. MICs were determined by agar or broth dilution methods. The stability of trospectomycin in the presence of an enzyme extract derived from spectinomycin-resistant Escherichia coli was determined. Trospectomycin was more active than spectinomycin (4- to 32-fold) against strains of numerous bacterial species, including staphylococci, streptococci, Haemophilus influenzae, Branhamella catarrhalis, Neisseria gonorrhoeae, Proteus species, Bacteroides species, Clostridium difficile, Clostridium species, and Chlamydia trachomatis. Trospectomycin demonstrated a moderate level of activity (comparable to that of spectinomycin) for most species of the family Enterobacteriaceae tested and was generally cross resistant with spectinomycin. Trospectomycin was susceptible to inactivation by crude enzyme preparations from spectinomycin inactivating strains of E. coli. Trospectomycin inhibited a variety of clinically important organisms, including agents of sexually transmitted diseases and pelvic inflammatory disease. Clinical studies with this novel aminocyclitol antibiotic are in progress. PMID- 2966607 TI - Low-level colonization of hospitalized patients with methicillin-resistant coagulase-negative staphylococci and emergence of the organisms during surgical antimicrobial prophylaxis. AB - By use of techniques that have been developed to detect small numbers of methicillin-resistant staphylococci, we cultured samples from the nares and subclavian and inguinal areas of 29 patients before and after cardiac surgery and 10 patients before and after coronary angioplasty. Methicillin-resistant coagulase-negative staphylococci were recovered before the surgical or angioplasty procedure from 74% of patients. The quantitative recovery of methicillin-resistant isolates before cardiac surgery or coronary angioplasty was compared with the number of methicillin-resistant staphylococci detected at the same site 3 days after the procedure. In cardiac surgery patients (who received antibiotic prophylaxis), 17 of the 28 sites (61%) in which low-level colonization with methicillin-resistant strains was detected preoperatively contained high levels of methicillin-resistant staphylococci postoperatively. In contrast, coronary angioplasty patients (who did not receive antibiotic prophylaxis) did not have any of the 14 sites containing low levels of methicillin-resistant strains before angioplasty emerge to harbor high levels of methicillin-resistant staphylococci after angioplasty. Methicillin-resistant coagulase-negative staphylococci from each site in which high levels of methicillin-resistant staphylococci emerged postoperatively were paired with preoperative isolates from the same site. Identical antibiograms and plasmid profile patterns were demonstrated for seven of the pre- and postoperative isolate pairs, suggesting that the high levels of methicillin-resistant coagulase-negative staphylococci detected on the skin or in the nares after cardiac surgery were derived from methicillin-resistant organisms present at the site preoperatively in much smaller numbers. PMID- 2966609 TI - In vitro drug susceptibility of Chlamydia sp. strain TWAR. AB - The in vitro susceptibilities of eight Chlamydia sp. strain TWAR isolates were tested against tetracycline, erythromycin, penicillin, ampicillin, sulfisoxazole, and a new drug, trospectomycin. The ranges of inhibitory concentrations of these antimicrobial agents, except for sulfonamide, were similar to those for Chlamydia trachomatis. Sulfisoxazole was not inhibitory at the highest nontoxic concentration tested. PMID- 2966610 TI - Bioavailability of oral trimetrexate in patients with acquired immunodeficiency syndrome. AB - The combination of the lipophilic antifolate trimetrexate and the rescue agent leucovorin has shown promise in the treatment of Pneumocystis carinii pneumonia in patients with acquired immunodeficiency syndrome. The pharmacokinetic behavior of trimetrexate administered either by intravenous bolus or orally was studied in six patients with acquired immunodeficiency syndrome with a reversed-phase high pressure liquid chromatography assay. The mean clearance following bolus injection was 38 ml/min per m2, with a range of 15 to 55 ml/min per m2. The postdistributive half-life ranged from 6 to 16 h. With oral administration, the mean bioavailability was 44% (range, 19 to 67%). An oral dose of 60 mg/m2 (162 mumol/m2) resulted in concentrations in plasma that approximated those achieved with a 30-mg/m2 (81-mumol/m2) intravenous dose. The toxicity of this combination regimen was minimal. It appears that the oral route is a practical route of administration for trimetrexate in patients with acquired immunodeficiency syndrome requiring long-term outpatient treatment or prophylaxis for P. carinii pneumonia. PMID- 2966611 TI - Acquired progressive lymphangioma as a flat erythematous patch on the abdominal wall of a child. AB - An erythematous patch was noted on the abdominal wall of an 8-year-old boy. The lesion showed a prolonged initial clinical course, followed by rapid later growth, finally reaching 3.7 X 7.0 cm in size over four years. Despite the harmless clinical appearance, the lesion was histologically characterized by tortuous vascular channels with some cellular atypia. Immunoperoxidase staining disclosed no factor VIII-related antigen or reaction to Ulex europaeus I lectin on tumor cells. There has been no recurrence three years after local excision. Although many features in our case resemble those reported in the literature under the term low-grade angiosarcoma, our preferred designation for such cases is acquired progressive lymphangioma, rather than angiosarcoma, because of their benign behavior. PMID- 2966612 TI - Changes in the transmural distribution of glucose-metabolizing enzymes across the left and right ventricular wall of rat heart during growth and ageing. AB - The age-related changes in the activities of five glucose-metabolizing enzymes (hexokinase, HK; glucose-6-phosphate dehydrogenase, G6P-DH; aldolase, ALD; phosphofructokinase, PFK; and lactate dehydrogenase, LDH) were investigated in the walls of left and right ventricles of rats of various age-groups (1-24 months). Age-related changes were found in the activities of all of the enzymes in both ventricles during growth (with significant decreases between 2 and 6 months of age) and in the levels of PFK and LDH in the left ventricle during ageing (with a significant increase between 12 and 24 months of age). The distribution of the enzyme activities across the wall of both ventricles was quite uniform in young, adult and mature rats (the distribution of G6P-DH activity in the left ventricle wall at 2 months of age was the only notable exception) but became non-uniform in the old rats with regard to G6P-DH, PFK, LDH and probably HK in the left ventricle and G6P-DH and HK in the right ventricle. These data support the hypothesis that alterations connected with ageing do not lead to a generalized decline of cardiac metabolic capacity, and that they are also the result of specific adaptive modifications, perhaps related to alteration in the distribution of the work load and/or of nutrition across the ventricular wall. PMID- 2966613 TI - Acute effects of maprotiline, doxepin and zimeldine with alcohol in healthy volunteers. AB - In a double-blind and cross-over trial, 12 healthy volunteers received single oral doses of maprotiline 75 mg, doxepin 25 mg, zimeldine 200 mg and placebo, alone and with alcohol (1 g/kg), at one-week intervals. Objective tests of performance (tracking, choice reaction, flicker fusion, body sway, nystagmus, Maddox wing) and ratings of subjective feelings were done before the drug intake (baseline) and 1 1/2, 3, 4 1/2, 6 and 7 1/2 hr after it. Maprotiline and doxepin proved subjectively sedative, whilst doxepin and zimeldine prolonged choice reaction time. An increase of alcohol effects was seen clearly after doxepin, to lesser extent after maprotiline and not at all after zimeldine. On the contrary, zimeldine antagonized alcohol-induced impairment in the tracking test. PMID- 2966614 TI - Cerebellar influence on the endorphinergic neural system. PMID- 2966615 TI - [Treatment of femoro-popliteal thrombosis by peripheral angioplasty. Value of the laser]. AB - In a series of 197 dilatations of the peripheral arteries performed over a 5-year period (1980-1985), 38 concerned complete thrombosis (less than 12 cm long) of the femoral and popliteal arteries. While in patients with femoral stenosis (n = 49) the primary success rate was 95 p. 100 with an annual recurrence rate of 8 p. 100, in patients with complete thrombosis the primary success rate was only 52 p. 100, and the recurrence rate was 30 p. 100 at 6 months. Such results would justify the use of new therapeutic methods. Percutaneous laser thermal angioplasty is now being evaluated, and it appears that the primary success rate is considerably increased by combined laser and balloon angioplasty. Recurrences raise a much more complex problem and require a multifactorial management with improved dilatation techniques, anti-platelet or anticoagulant treatment, abstention from smoking and close surveillance. PMID- 2966616 TI - Families as caregivers: the limits of morality. AB - There is a developing trend to look to families to provide care and support of those in need of rehabilitation. A widespread assumption is that family care is superior care and, with modest degrees of support, families can provide that care. Yet we may question the ethical limits of the obligation of a family or family member to provide care when the demands are severe. Psychologic and moral problems that confront caregivers are examined. In some cases caregivers must sacrifice their present and future welfare. That the moral claim made upon them may seem a justifiable one in many respects does not mean that it will be endurable; that it is endurable does not mean that it is justifiable. The problem is exacerbated by lack of a supportive culture, one that rewards and honors those who take on heroic duties. Caregivers may be socially isolated. If heroic demands are to be made on family members, a richer moral culture is required, not just the provision of improved social services. PMID- 2966617 TI - Admission to rehabilitation centers: selection of patients. AB - Patients are selected for admission to rehabilitation centers. Some patients are too sick or medically unstable to treat; others' disabilities are irremediable. Resources should not be expended upon patients who will not benefit from treatment. The current process of patient selection requires providers to consider both medical and nonmedical factors. Medical factors include diagnosis, prognosis, secondary complications, functional performance and prognosis, and ability to learn. Nonmedical assessment includes social, vocational, financial, and personal factors. There is a potential for subjectivity in the way these variables are considered. Practitioners may be unaware of bias in their decision making. Patients may be disadvantaged by lack of family or financial resources. Both providers and patients suffer when providers are forced by expectations of utilitarianism to deny treatment to deserving patients. It would be helpful if patients and families were informed about the process of selection of rehabilitation candidates and given the right of redress if rejected. Guidelines delineating standards for admission to rehabilitation centers could assist patient selection. Finally, rehabilitationists must seek to further demonstrate the efficacy of their treatment in order to better identify persons who will benefit from rehabilitation services. PMID- 2966618 TI - Justice in the allocation of public resources to disabled citizens. AB - This paper examines several crucial questions concerning justice in the allocation of public resources to disabled citizens. It challenges the current failure to means-test certain programs, and it argues that fundamental choices about levels of funding and about priorities in nonentitlement programs cannot be justified without a better theory of social justice. PMID- 2966619 TI - Dystonia musculorum deformans: three cases treated on a rehabilitation unit. AB - Dystonia musculorum deformans is a rare involuntary movement disorder of unknown etiology. We treated three brothers in one family on our rehabilitation unit, resulting in definite improvement in their ability to walk and control the dystonic movements, and in their speech and swallowing. All three benefited from orthoses to prevent equinovarus and maintain ankle stability. Intense concentration, such as needlepoint, seemed to reduce the amount of extra motion. Performing another activity during dystonic movements, such as pointing to a communication board, also seemed to give some relief. A special seating system using a collapsible wheelchair, a low center of gravity, and antitip wheels reduced one brother's frequency of falling. Swallowing finely cut foods was not a problem with any of our patients, although one had to be instructed in a special technique. An electronic communication board was very helpful. Our success in these cases should encourage others working in rehabilitation to consider such patients and to apply the principles of rehabilitation, particularly the total team approach, in their care. PMID- 2966620 TI - [The chalone-antichalone system, and mechanisms regulating granulocytopoiesis]. AB - The normal physiological balance between granylocytic chalone and antichalone is shown to be significantly upset where an inflammatory focus is present, with segmented neutrophils activated by cell breakdown products producing, for the most part, a stimulator of cell proliferation. It is also shown that the production of tissue-specific regulators in inflammation depends on the functional state of the entire mature neutrophil pool. The mechanisms of intrasystemic granulocytopoiesis regulation have been unraveled. PMID- 2966621 TI - Enzyme-histochemical differences in fibre-type between the human major and minor zygomatic and the first dorsal interosseus muscles. AB - Human masticatory muscles, innervated by the trigeminal nerve, differ in fibre type composition from limb and trunk muscles, but the anterior and the posterior belly of the human digastric muscle, innervated by the trigeminal and facial nerves, respectively, do not. The major and minor zygomatic muscles from adult males, which originate from the second branchial arch and are supplied by the facial nerve, were analysed enzyme-histochemically and compared with the first dorsal interosseus hand muscle, which has spinal innervation and, like the masticatory and facial muscles, a large cortical representation. Both zygomatic muscles had a marked predominance of type II fibres, the minor one having the largest proportion (89.1 per cent) ever reported in human skeletal muscle. Besides type I, IIA, IIB, and a few type IIC fibres, there was a large group with an ATPase reaction at pH 4.6, between that of type IIA and type IIB, and termed IIAB. This fibre-type profile may reflect a special isomyosin composition. Type I and II fibres were of about equal diameter, corresponding to that of type I fibres in the masticatory muscles. Individual and intra-muscular variability in fibre size and shape was considerable. The unusually high frequency of type II fibres in the zygomatic muscles suggest that they have fast-contraction properties and relatively large motor units, and therefore are poorly adapted to finely-graded movements. The absence of muscle spindles supports this view. The hand muscle had a chequer-board pattern of type I, IIA and IIB fibres, similar to that of large limb and trunk muscles, with no difference between its two heads.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966623 TI - Stress responses in lambs docked and castrated surgically or by the application of rubber rings. AB - A comparative study has been made in lambs 3 to 6 weeks of age of the behavioural responses and changes in plasma immunoreactive beta-endorphin (ir beta-endorphin) and cortisol after docking or docking plus castration by the application of rubber rings or by surgery. The use of rubber rings on lambs at this age was characterised by very agitated behaviour indicative of considerable distress for a period of up to 1 h. In contrast, surgery was accompanied by some post operative shock exhibited by reduced exploratory and social behaviour, at least in the lambs exposed to docking plus castration. In the latter group there were highly significant increases in both plasma ir beta-endorphin and cortisol concentrations that may be consistent with the induction of stress-induced analgesia. We conclude that surgery caused less distress than the rubber rings, and is therefore preferable for docking and castration of lambs 3 to 6 weeks of age. PMID- 2966624 TI - The post-accident syndrome: variations in the clinical picture. AB - Three hundred and twenty-seven subjects involved in civil accident litigation and referred by solicitors for either plaintiff or defendant were examined. A precise psychiatric diagnosis was applicable in only a minority of subjects, although psychiatric symptoms, including pain, anxiety and depression, were prominent. Overall the clinical picture was an amorphous one. Cluster analysis was performed to examine a variety of clinical, demographic and historical variables. This showed four stable groups, which are here described as: stoic, depressive, phobic motor accident and prior claimants. It is suggested that the phenomenological approach taken here, paying particular attention to clinical sub-groups, may be a more useful route towards the understanding and treatment of post-accident psychiatric disturbance than is the existing but confused approach, in which motivation has been a prime focus of interest. The particular sub-groups demonstrated here, if replicated, could form the basis for such an approach. PMID- 2966625 TI - A legal viewpoint on the relevance of phenotypic variation to risk assessments. PMID- 2966626 TI - A novel biological effect of atrial natriuretic hormone: inhibition of mesangial cell mitogenesis. AB - We have investigated the effect of atrial natriuretic hormone on serum-induced mitogenesis in cultured rat mesangial cells. Synthetic peptides, atriopeptin 28 and atriopeptin 24, dose-dependently decreased thymidine incorporation, with a half-maximal effect at approximately 1 nM and a maximal inhibition of approximately 60%. Moreover, atriopeptin 28 significantly decreased the clonal proliferation of mesangial cells. Atriopeptin 28 also decreased resting cytosolic Ca but had no effect on the increase induced by serum, relative to the lower baseline established by atriopeptin 28. Nevertheless, the overall effect of atriopeptin 28 on Ca was to attenuate the serum-induced increase, relative to the original resting level. These results therefore provide evidence for a novel biological effect of atrial natriuretic hormone and suggest that the antimitogenic effect may be mediated by atriopeptin-induced alterations of intracellular Ca dynamics. We speculate that atrial natriuretic hormone may be a modulator of mesangial cell mitogenesis in vivo. PMID- 2966627 TI - Dopamine D1 receptors with enhanced agonist affinity and reduced antagonist affinity revealed by chemical modification. AB - In order to investigate the possibility that there may be two conformationally distinct dopamine D1 binding sites, the effect of lysine-modifying agents on striatal dopamine D1 receptors was investigated. Treatment with the distilbene derivative, 4,4'-diisothiocyanostilbene-2,2'-disulfonate, (DIDS), resulted in an irreversible D1 receptor inactivation that was associated with a 70% loss of binding sites. The remaining DIDS-insensitive sites displayed both a decreased affinity (approximately 5 fold) for the D1 antagonist SCH-23390 and an enhanced affinity of dopaminergic agonists (approximately 10 fold) for the agonist high affinity form of the receptor. Pretreatment with Gpp(NH)p, a non-hydrolysable guanine nucleotide, prevented the formation of the agonist high-affinity form, indicating that these sites are G-protein-linked. Prior occupancy of D1 receptors with dopaminergic agonists and antagonists afforded no protection against DIDS inactivation, suggesting that a site outside the ligand binding subunit of the D1 receptor was modified. Taken together, these data suggest that [3H]SCH-23390 labels two conformationally distinct populations of dopamine D1 receptors. PMID- 2966628 TI - Acute, subchronic and chronic toxicity of the new angiotensin converting enzyme inhibitor ramipril. AB - Acute, subchronic and chronic toxicity studies of 2-[N-[(S)-1-ethoxycarbonyl-3 phenylpropyl]-L-alanyl]-(1S, 3S, 5S)-2-azabicyclo[3.3.0]octane-3-carboxylic acid (ramipril, Hoe 498) were conducted in mice, rats, dogs and monkeys. The acute toxicity in mice, rats, and dogs after oral and intravenous administration is very low. There were no significant differences in LD50 values between male and female animals. Treatment periods up to 6 months in rats and monkeys and up to 12 months in dogs revealed consistent compound-related effects with respect to the kidneys and the hemopoietic system probably due to exaggerated pharmacological activity. Species-specific lesions in the gastric fundal mucosa were noted exclusively in rats. No adverse effects were seen at doses of 0.25 mg/kg/d in rats, 2.5 mg/kg/d in dogs, and 0.5 mg/kg/d in monkeys. It is concluded that ramipril can be safely used in humans. PMID- 2966629 TI - Immunopharmacological actions of the new antiallergic drug butyl 3'-(1H-tetrazol 5-yl)oxanilate. 1st communication: effects on type I to type IV allergic reactions in animal models. AB - The effects of butyl 3'-(1H-tetrazol-5-yl)oxanilate (WP-833), a new antiallergic drug, on type I to type IV allergic reactions were investigated by employing various animal models. WP-833 (i.v. and p.o.) dose-dependently inhibited homologous or heterologous passive cutaneous anaphylaxis (PCA) mediated by rat or mouse immunoglobulin E (IgE) in rats. Homologous PCA caused by guinea pig IgE was also inhibited by WP-833. In addition, WP-833 had inhibitory actions upon homologous PCA induced by rat or guinea pig IgG. However, WP-833 showed no inhibition of rat skin reactions caused by histamine, serotonin and bradykinin, contrasting with the inhibition of prostaglandin E1-induced skin reaction. Furthermore, both adrenalectomy and propranolol treatment exerted no influences on the inhibition of IgE-mediated homologous PCA in rats by WP-833. In contrast to above findings demonstrating that WP-833 clearly inhibited type I allergic reaction, systemic Forssman shock in guinea pigs and reversed cutaneous anaphylaxis in rats (type II), passive Arthus reaction in rats (type III), and contact dermatitis and tuberculin reaction in mice (type IV) were unaffected by WP-833 even in higher doses than in those capable of completely inhibiting type I allergic reaction. PMID- 2966630 TI - Self-care practices to prevent low back pain. PMID- 2966631 TI - Effects of long-term seropositivity to human immunodeficiency virus in a cohort of homosexual men. AB - The long-term effects of HIV infection were evaluated by comparing data from two visits a mean of 18 months apart in groups of 148 persistently seropositive and 287 persistently seronegative homosexual men. At each visit, the seropositive men exhibited lower CD4 counts, CD4/CD8 ratios, hemoglobin concentrations and lymphocyte counts, and higher C1q binding, IgG and IgA levels. More important, the decline of the CD4/CD8 ratio and the rise of the C1q binding, IgG and IgA, progressed significantly in the seropositive group between visits. Seropositive men were at elevated risk of developing constitutional symptoms and generalized lymphadenopathy. An association was present between development of symptoms and inversion of the CD4/CD8 ratio. The 11 seropositive men who have progressed to AIDS had lower CD4 counts and CD4/CD8 ratios, and higher C1q binding, IgG and IgA, than 134 seropositive AIDS-free men a mean of 21.4 months prior to diagnosis. The AIDS group demonstrated greater decline between visits in the CD4 count, hemoglobin and white blood count (WBC) than the seropositive AIDS-free group. The present data document the long-term effects of HIV infection in a seropositive cohort and suggest the possibility of a subgroup particularly susceptible to the progressive effects of HIV that precede the development of the acquired immunodeficiency syndrome (AIDS). PMID- 2966632 TI - Decreased expression of the C3b/C4b complement receptor (CR1) in AIDS and AIDS related syndromes correlates with clinical subpopulations of patients with HIV infection. AB - Expression of the C3b/C4b receptor (CR1) was studied on erythrocytes of 153 individuals infected with HIV and 104 age-matched normal individuals by measuring the uptake of 125I-labelled monoclonal anti-CR1 antibody. The mean number of CR1 sites on erythrocytes of asymptomatic seropositive individuals (822 +/- 270; mean +/- s.d.) and of patients with persistent generalized lymphadenopathy (PGL; 775 +/- 320) did not differ significantly from that of normal subjects. The number was significantly lower in patients with AIDS-related complex (ARC; 543 +/- 233; P less than 5 x 10(-3)) and further decreased in patients with AIDS (442 +/- 271; P less than 1 x 10(-4)), whether they presented with Kaposi's sarcoma (KS) or opportunistic infections. An additional finding was that of decreased expression of antigenic and functional CR1 in neutrophils from patients with AIDS, as assessed by radioimmunoassay of CR1 in detergent-solubilized cells and the capacity of intact cells to form rosettes with C3b-coated erythrocytes. Low numbers of CR1 on cells from patients with AIDS were not due to occupation of the receptor by C3 fragments on immune complexes. The correlation that was observed between decreased numbers of CR1 on erythrocytes and clinical subpopulations of symptomatic HIV-infected patients suggests that CR1 expression on erythrocytes may represent a valuable marker of the severity and natural history of HIV associated disease. PMID- 2966633 TI - Elevated soluble interleukin-2 receptor levels in serum of human immunodeficiency virus infected populations. AB - Soluble interleukin-2 receptor (SIL-2R) levels in sera were quantitated in asymptomatic intravenous drug abusers (IVDA) and in patients with lymphadenopathy or AIDS. The mean SIL-2R level in serum of normal controls was 158 +/- 19 compared to 368 +/- 35 U/ml in serum of HIV-seronegative asymptomatic IVDA. The mean SIL-2R in serum of HIV-seropositive asymptomatic IVDA was 609 +/- 85 U/ml and in patients with lymphadenopathy was 745 +/- 79 U/ml. In addition, AIDS patients with Pneumocystis carinii pneumonia, Kaposi's sarcoma, or both had elevated mean levels of SIL-2R values with a broad range. This elevated level of SIL-2R may reflect excessive cell surface IL-2R expansion by the infected cells. PMID- 2966634 TI - Obesity and postmenopausal bone loss: the influence of obesity on vertebral density and bone turnover in postmenopausal women. AB - This study was undertaken to evaluate the effect of obesity on the postmenopausal bone mass. Bone mineral density, measured by dual photon absorptiometry of the lumbar spine, serum osteocalcin (OC), fasting urinary calcium to creatinine (Ca:Cr), serum estradiol (E2) dehydroepiandrosterone (DHA) and testosterone (T) were measured in 176 women aged 45-71 years. Women were divided into four groups according to their menopausal status and their weight: 49 perimenopausal, 28 obese perimenopausal, 49 obese postmenopausal. Within each population (perimenopausal and postmenopausal), mean age was the same, only weight was significantly different (p less than 0.0001). For the two groups of postmenopausal women mean interval since menopause (YSM) was the same (5.8 +/- 3 and 5.4 +/- 5 yr). Comparison between groups revealed a significant effect of menopausal status and obesity on BMD and bone turnover. As compared to perimenopausal women, BMD was lower, OC and Ca: Cr higher only in nonobese postmenopausal women. E2, T, DHA did not differ between the two groups of postmenopausal women. The results of this study suggest that even moderate obesity can play a protective role on postmenopausal bone loss. PMID- 2966635 TI - Chronic instrumentation techniques for haemodynamic measurements. AB - Research in chronically instrumented experimental animals has shown basal anaesthesia may cause a considerable disruption of the autonomic control of the cardiovascular system, and a profound change in basal haemodynamics, myocardial function and coronary haemodynamics--variables which are of fundamental importance in studying the impact of various anaesthetics on the cardiovascular system. It is, therefore, important to make sequential haemodynamic measurements which enable comparisons to be made of the conscious v. the anaesthetized state. The present article describes some of the chronic instrumentation techniques used to study the effects of anaesthetic agents on overall haemodynamics, coronary blood flow and regional myocardial function in comparison with the conscious state. It is suggested that the effects of anaesthesia on the integrative control of the circulation are not sufficiently recognized. PMID- 2966636 TI - A general practice study investigating the effect of minocycline (Minocin) 50 mg bd for 12 weeks in the treatment of acne vulgaris. PMID- 2966638 TI - Localization and chemical synthesis of fibronectin peptides with melanoma adhesion and heparin binding activities. AB - Tumor cell adhesion to the extracellular matrix is an important consideration in tumor metastasis. Recent results show that multiple adhesion-promoting domains for melanoma cells can be purified from proteolytic digests of fibronectin [McCarthy, J. B., Hagen, S. T., & Furcht, L. T. (1986) J. Cell Biol. 102, 179 188]. Monoclonal antibodies were generated against a tryptic/catheptic 33K heparin binding fragment of fibronectin derived from the carboxyl terminal of the A chain. This region contains a tumor cell adhesion-promoting domain(s). The amino-terminal sequence was determined for this fragment, as well as a tryptic 31K fragment which is located to the carboxyl-terminal side of the 33K heparin binding fragment in A chains of fibronectin. The partial sequence data demonstrate that arginyl-glycyl-aspartyl-serine (RGDS) or the related arginyl glutamyl-aspartyl-valine (REDV) is not present in the 33K heparin binding fragment, confirming earlier results which demonstrated that cells adhere to this fragment by an RGDS-independent mechanism. Two monoclonal antibodies, termed AHB 1 and AHB-2, recognized epitopes common to heparin binding fragments derived from the carboxyl terminus of both the A and B chains of fibronectin. Monoclonal antibody AHB-2 inhibited melanoma adhesion to the 33K heparin binding fragment of fibronectin in a concentration-dependent manner, whereas monoclonal antibody AHB 1 had no effect on adhesion to this fragment. Neither monoclonal antibody inhibited adhesion to intact fibronectin. However, monoclonal AHB-2 potentiated the inhibitory effect of suboptimal levels of exogenous RGDS on cell adhesion to intact fibronectin. AHB-2 recognized an epitope common to both the A- and B-chain carboxyl-terminal heparin binding region of fibronectin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966637 TI - Recognition and repair of the CC-1065-(N3-adenine)-DNA adduct by the UVRABC nucleases. AB - The recognition and repair of the helix-stabilizing and relatively nondistortive CC-1065-(N3-adenine)-DNA adduct by UVRABC nuclease has been investigated both in vivo with phi X174 RFI DNA by a transfection assay and in vitro by a site directed adduct in a 117 base pair fragment from M13mp1. CC-1065 is a potent antitumor antibiotic produced by Streptomyces zelensis which binds within the minor groove of DNA through N3 of adenine. In contrast to the helix-destabilizing and distortive modifications of DNA caused by ultraviolet light or N-acetoxy-2 (acetylamino)fluorene, CC-1065 increases the melting point of DNA and decreases the S1 nuclease activity. Using a viral DNA-Escherichia coli transfection system, we have found that the uvrA, uvrB, and uvrC genes, which code for the major excision repair proteins for UV- and NAAAF-induced DNA damage, are also involved in the repair of CC-1065-DNA adducts. In contrast, the uvrD gene product, which has been found to be involved in the repair of UV damage, has no effect in repairing CC-1065-DNA adducts. Purified UVRA, UVRB, and UVRC proteins must work in concert to incise the drug-modified phi X174 RFI DNA. Using a site-directed and multiple CC-1065 modified (MspI-BstNI) 117 base pair fragment from M13mp1, we have found that UVRABC nuclease incises at the eighth phosphodiester bond on the 5' side of the CC-1065-DNA adduct on the drug-modified strand.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966639 TI - Further characterization of DNA helicase activity of mouse DNA-dependent adenosinetriphosphatase B (DNA helicase B). AB - The DNA helicase activity of DNA-dependent ATPase B purified from mouse FM3A cells [Seki, M., Enomoto, T., Hanaoka, F., & Yamada, M. (1987) Biochemistry 26, 2924-2928] has been further characterized. The helicase activity was assayed with partially duplex DNA substrates in which oligonucleotides to be released by the enzyme were radiolabeled. Oligonucleotides with or without phosphate at the 5' termini or with a deoxy- or dideoxyribose at the 3'-terminal nucleotides were displaced by this enzyme with essentially the same efficiency and with the same ATP (and dATP) and Mg2+ requirements. Thus, there was no strict structure requirement for both ends of duplex regions of substrates to be unwound by the enzyme. Shorter strands were released more readily than longer strands up to the length of 140 bases. The attachment of the enzyme to a single-stranded DNA region was a prerequisite for the neighboring duplex to be unwound; the enzyme-catalyzed unwinding was inhibited competitively by the coaddition of single-stranded DNAs which act as cofactors of the ATPase activity. Their activities as the inhibitor of helicase were well correlated with those as the cofactor of ATPase. The helicase B was found to migrate along single-stranded DNA in the 5' to 3' direction by the use of single strands with short duplex regions at both 3' and 5' ends as substrate. A possible role of this enzyme in DNA replication in mammalian cells is discussed. PMID- 2966640 TI - Patterns of proteolytic cleavage and carbodiimide derivatization in sarcoplasmic reticulum adenosinetriphosphatase. AB - Two series of experiments were carried out to characterize (a) peptide fragments of sarcoplasmic reticulum (SR) ATPase, based on proteolysis with different enzymes and distribution of known labels, and (b) specific labeling and functional inactivation patterns, following ATPase derivatization with dicyclohexylcarbodiimide (DCCD) under various conditions. Digestion with trypsin or chymotrypsin results in the initial cleavage of the SR ATPase in two fragments of similar size and then into smaller fragments, while subtilisin and thermolysin immediately yield smaller fragments. Peptide fragments were assigned to segments of the protein primary structure and to functionally relevant domains, such as those containing the 32P at the active site and the fluorescein isothiocyanate at the nucleotide site. ATPase derivatization with [14C]DCCD under mild conditions produced selective inhibition of ATPase hydrolytic catalysis (EP + H2O in equilibrium E + Pi) without significant incorporation of the 14C radioactive label. This effect is attributed to blockage of catalytically active residues by reaction of the initial DCCD adduct with endogenous or exogenous nucleophiles. ATPase derivatization with [14C]DCCD under more drastic conditions produced inhibition of calcium binding, 14C radioactive labeling of tryptic fragments A1 and A2 (but not of B), and extensive cross-linking. Intermolecular and, to some extent, intramolecular cross-linking were prevented by exogenous nucleophiles. The presence of calcium during derivatization prevented functional inactivation, radioactive labeling of fragment A2, and internal cross-linking of fragment A1. It is proposed that both A1 and A2 fragments participate in formation of the calcium binding domain and that the labeled residues of fragment A2 are directly involved in calcium complexation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966642 TI - Olfactory deficits and Down's syndrome. PMID- 2966643 TI - (18O)quinolinic acid: its esterification without back exchange for use as internal standard in the quantification of brain and CSF quinolinic acid. AB - In the process of developing a high-sensitivity negative chemical ionization gas chromatographic/mass spectrometric assay for brain and cerebrospinal fluid (CSF) levels of quinolinic acid (QUIN, 2,3-pyridine dicarboxylic acid), (18O4)QUIN was prepared. Its properties as an internal standard were compared with those of the structural isomer 2,4-pyridine decarboxylic acid (2,4-PDC) previously used by others. All oxygen atoms in QUIN were labeled by heating in 3 N HCl/(18O)water for 48 h at 80 degrees C. Back-exchange of (18O4)QUIN was prevented during derivatization to an electron-capturing dihexafluoropropanol ester by using trifluoroacetylimidazole as catalyst instead of perfluroacyl anhydrides. When mixtures of QUIN and (18O4)QUIN and/or 2,4-PDC were followed through a procedure to isolate and quantify brain and CSF QUIN, the variability in the ratio of QUIN:2,4-PDC was greater than for QUIN: (18O)QUIN. We conclude that (18O)QUIN is the preferred internal standard in gas chromatographic/mass spectrometric quantification of brain and CSF QUIN, and that (18O)-labeled carboxylic acids may be esterified effectively without back-exchange using acylimidazole reagents. PMID- 2966641 TI - Secretion of the lysosomal acid triacylglycerol hydrolase precursor by J774 macrophages. AB - J774, thioglycollate-elicited mouse peritoneal and BCG-induced rabbit alveolar macrophages all contain high levels of a triacylglycerol hydrolase (EC 3.1.1.3) (TGase) with optimal activity at pH 6.5. The J774 macrophages, a cell line deficient in the calcium-independent mannose 6-phosphate receptor, were found to secrete large quantities of the TGase into the culture medium. In contrast, mouse peritoneal and rabbit alveolar macrophages, which are both mannose 6-phosphate receptor-competent cell types, secreted much lower amounts of neutral TGase. The enzyme was localized in the lysosomes of rabbit alveolar macrophages. Addition of 25 mM NH4Cl induced a 6-fold increase in TGase secretion by alveolar macrophages, while 50 mM NH4Cl induced a 12-fold increase in TGase secretion. NH4Cl had no effect on TGase secretion by J774 macrophages. The TGase secreted by J774 macrophages was internalized by I-cell disease fibroblasts, increasing the cellular content of TGase 10-fold after 8 h. Internalization was inhibited 70% by the addition of 2 mM mannose 6-phosphate to the culture medium, but was not affected by 2 mM mannose or glucose 6-phosphate. After internalization, the neutral TGase was converted to a TGase with a pH optimum of 5.1. These data are consistent with the spontaneous release of a lysosomal enzyme precursor from a calcium-independent mannose 6-phosphate receptor-deficient cell line, indicating that the neutral TGase previously reported in several types of macrophages may be the precursor of the lysosomal acid TGase. PMID- 2966644 TI - Anti-bromodeoxyuridine monoclonal antibody: an alternative tool for the identification of replicated DNA at the electron microscope level. AB - A new method for identifying the replicated DNA at the electron microscope level is described. Cells were first exposed in vitro to 5-bromodeoxyuridine (BUdR) in conjunction with 5-fluorodeoxyuridine (FUdR) and BUdR incorporated into DNA was then detected on Lowicryl-embedded sections by immunogold technique using a monoclonal anti-BUdR antibody. After using this method, chromatin and chromosomes are strongly labelled. PMID- 2966645 TI - Disabled living centres. PMID- 2966646 TI - Benzydamine oral rinse and rash. PMID- 2966647 TI - Severe toxic erythema caused by diltiazem. PMID- 2966648 TI - Postural effects on F wave parameters in lumbosacral root compression and canal stenosis. AB - The effects of standing, and lying with the back extended, on F wave minimal latency and F chronodispersion were studied in patients with lumbar canal stenosis, lumbosacral root compression syndromes, and low back pain. Standing for 3 min produced increased F chronodispersion in lumbar canal stenosis and root compression syndromes but had less effect on the F minimal latency. In patients with low back pain these variables were unchanged by these postural manoeuvres. These observations illustrate the value of adapting electrophysiological investigations to those clinical features that induce symptoms in these disorders. PMID- 2966649 TI - Cytochemical localization of Ca2+-ATPase activity in peripheral nerve. AB - We used an electron microscopic cytochemical method to determine the localization of Ca2+-ATPase in rat peripheral nerve. We found that reaction product occurred along most cytoplasmic membranes in the dorsal root ganglia (DRG). Unmyelinated axons demonstrated reaction product on the axolemma diffusely along their length. Myelinated fibers, in contrast, had reaction product limited to the axolemma in the paranodal region. Internodal axolemma never showed reaction product and nodal axolemma was only occasionally stained, usually in sections reacted for the maximum times. Schwann cell plasma membranes uniformly showed reaction product. The restricted localization of Ca2+-ATPase to the paranodal region of myelinated fibers suggests that calcium efflux may occur principally at those sites. PMID- 2966650 TI - Can the bad guys be defeated? PMID- 2966651 TI - Host response to myeloma: I. Induction of cytotoxic and suppressor T cells by in vivo immunization with MOPC 104E plasmacytoma. AB - Spleen cells from BALB/c mice immunized with mitomycin C-treated MOPC 104E plasmacytoma cells demonstrated negligible cytotoxic activity (less than 10% specific cytotoxic activity) in the 51Cr release assay. These cells exhibited increased cytotoxic activity when they were secondarily sensitized in vitro with mitomycin C-treated MOPC 104E cells. Spleen cells from normal mice showed tumor specific cytotoxic activity when cocultured with mitomycin C-treated tumor cells at the optimal responder to stimulator ratio of 25:1. The level of cytotoxic activity obtained by in vivo primed and secondarily in vitro sensitized spleen cells did not exceed the level of activity obtained by in vitro primary sensitized cells. Significant suppression of the cytotoxic activity of in vitro primary sensitized cells was observed when cocultured with in vivo primed spleen cells during primary sensitization in vitro at a responder to suppressor cell ratio of 1:1. Suppressor cells of in vivo primed mice were removed by treatment with anti-Thy 1.2 and complement. These results suggest that spleen cells from in vivo primed mice consisted of at least two subpopulations of cells, a cytotoxic (prekiller) and suppressor T cells. Attempts to induce cytotoxic cells in vivo might have failed because of the appearance of suppressor T cells. PMID- 2966652 TI - [Expression of the human insulin gene in transgenic mice]. AB - Transgenic mice which specifically express the human insulin gene in the pancreatic beta cells have been obtained. Expression is demonstrated by the presence in the serum of human insulin (determined by a radioimmunoassay for the human C-peptide), the level of which increases upon glucose stimulus, and by the presence of human insulin mRNA in pancreas, but not in the other tissues tested. This result indicates that nucleotide sequences necessary for tissue-specific expression are within the transferred gene and/or in its vicinity. It also suggests that the regulatory molecules are quite similar if not identical in human and mouse. PMID- 2966653 TI - [Effects of the growth factors EDGF I and EDGF II on wound healing of the corneal epithelium in rabbits]. AB - EDGF I and II, the retinal forms of basic and acid FGF, isolated from adult bovine retina, are both able to increase the rate of wound healing of rabbit corneal epithelium. PMID- 2966654 TI - [Gene expression of D-beta-hydroxybutyrate dehydrogenase. III. Molecular cloning of a DNAc]. AB - In order to continue the molecular studies of D-beta-hydroxybutyrate dehydrogenase (BDH) undertaken in our laboratory for several years, we have initiated a genetic approach which consists in the BDH cDNA cloning from a rat liver cDNA library. The immunoscreening method allowed to isolate a clone which exhibits a DNA insert shorter than the expected full length BDH cDNA. PMID- 2966655 TI - [Sulfation and mitogenic activities of growth factors in human serum: importance of compounds of molecular weight lower than 1,000 in the global expression of activity of this serum on 3 different cellular types]. AB - A human serum ultrafiltrate contains compounds needed for a maximal expression of sulfation and mitogenic activities of the corresponding retentate. These low molecular weight (less than 1,000) molecules have no effect, by their own, on 35SO4(2-) uptake in chick embryo cartilage, but show a significant synergistic effect with serum growth polypeptides. Tested alone, they display a slight mitogenic activity as measured by 3H-thymidine incorporation in chick embryo fibroblasts or in lymphocytes activated by phytohemagglutinin. Here they also act in a synergistic way on mitogenic activities of growth factors from human retentate. A fraction (P2B), partially purified from human plasma ultrafiltrates, produces the same synergistic response with human retentate in the three cellular systems (cartilages, fibroblasts, activated lymphocytes). However, concentrations which lead to optimum responses are very different according to the cell type studied. These results suggest the existence of several compounds which can each act specifically on a cellular system. PMID- 2966656 TI - [Effects of the level of polyunsaturated fatty acids of the diet on the functioning and depletion of energy substrates in the rat heart during perfusion by left atrial route]. AB - The effects of semi-liquid diets containing 6.6% in weight of refined sunflower seed oil (SSO) or hydrogenated coconut oil (HCO) on cardiac endogenous substrates and functional parameters of rats hearts were compared to a standard laboratory chow during seven days. No difference appeared for cardiac glycogen and lipid constituents. Cardiac performance, measured through left atrial perfusion was enhanced by SSO diet and HCO one altered it. A significative phospholipid depletion appeared during the 45 minutes perfusion only in the HCO group. PMID- 2966657 TI - [Effects of the ingestion of oxidized fish oils on hepatic microsomal enzyme activities and membrane fluidity in rats. Influence of tocopherol overload]. AB - Rats fed a dietary peroxidized fish oil showed an increase in cytochrome P-450 content and ethoxy-coumarin deethylase (ECDE) activity in liver microsomes. Administration of DL-alpha-tocopherol led to different effects according to the extent of the peroxidation in the fish oil. In rats fed a de-peroxidized oil, the inductive effect of phenobarbital on UDP-glucuronosyltransferase (UDGPT) activity was depressed by tocopherol. By the same time, induction of P-450 and ECDE remained unchanged, that of epoxide hydrase slightly increased. By contrast tocopherol strongly potentiated the inductive effect of phenobarbital toward UDPGT activity (group I substrates) in rats fed the peroxidized fish oil. The modification of the inductive effect of phenobarbital in combination with tocopherol on UDPGT activities was concomitant with an increase in seric transaminase activity and with a reverse effect as revealed from the study of the rate of fluorescent probes penetration in microsomes. The possible toxicity of the strong dose of tocopherol is discussed. PMID- 2966658 TI - [Substrate specificity and kinetic properties of neutral maltase of human granulocytes]. AB - A neutral maltase immunologically similar to this of kidney exist in human granulocytes. We have studied some kinetic properties of this enzyme on a microsomal fraction of granulocytes. Its optimal pH is very closed of 6.8 and this enzyme, highly specific for maltose, hydrolysis very weakly the nigeriosis. Maltotriose, maltotetraose and maltopentanose are inhibitors of this enzyme, which is not inhibited by all disaccharides studied. PMID- 2966659 TI - [Autonomic cardiovascular regulation and permanent catheterization in normotensive conscious (WKY) and spontaneously hypertensive (SHR) rats]. AB - In cardiovascular research, methods of indwelling catheterism have been frequently described. In the present work, we used normotensive and SHR rats to compare carotid catheterism to left ventricular catheterism, this last method being proposed by some investigators for cardiac index measurement. Our results of plasma catecholamines and autonomous nervous system activity show that ventricular catheterism results in an important disturb in cardiovascular regulation and question its validity to study cardiac function. PMID- 2966660 TI - [Effects of nicardipine on the spontaneous beat of isolated atria of guinea pigs]. AB - Nicardipine was found to produce a concentration-dependent depression of the isometric contraction of the isolated, spontaneously beating atria of the guinea pig. It also depressed the atrial rate of the isolated, spontaneously beating atria of the guinea-pig. The effect of the increasing concentrations of nicardipine on the heart rate was negligibly weaker than its effect on the isometric contraction. A time-dependent depression of the isometric contraction and the atrial rate after the addition of a single dose of nicardipine was also found up to the 10th min. Calcium almost completely, isoprenaline completely and aminophylline partially antagonized the depressive action of nicardipine on the isometric contractility of the atria. Only isoprenaline antagonized the negative chronotropic action of higher doses of nicardipine. It is possible that these substances restore the contractility by compensating the calcium balance, previously changed by nicardipine, or by producing an increase in the intracellular cAMP content (isoprenaline and aminophylline). PMID- 2966661 TI - Keratan sulfate content in the superficial and deep layers of osteophytic and nonfibrillated human articular cartilage in osteoarthritis. AB - Very thin slices of the superficial and deep layers of osteophytic and apparently normal articular cartilage from six human osteoarthritic femoral heads were digested with papain. The digests were analyzed for keratin sulfate content using an enzyme-linked immunosorbent assay (ELISA) with inhibition step, and for chondroitin sulfate and collagen contents using biochemical assays. Although there were marked differences in Safranin-O staining of the superficial and deep layers of osteophytic cartilage, these two layers had identical high ratios of chondroitin sulfate/collagen. Keratan sulfate was present only in small amounts in osteophytic cartilage. However, the deeper layer contained significantly more of this glycosaminoglycan. The deeper layer of articular cartilage contained approximately twice as much chondroitin sulfate and six times more keratan sulfate relative to collagen than the superficial layer. The results of this study showed that this new sensitive approach, which requires as little as 200 micrograms wet cartilage as starting material, provides important qualitative and quantitative information about the major constituents of the matrix. PMID- 2966662 TI - Carboxyhemoglobinemia, polycythemia, cardiomegaly, and cardiovascular function in the anesthetized rat. AB - The effects of carbon monoxide (CO), polycythemia (PC), and cardiomegaly (CM) on cardiovascular function were investigated in adult rats in which the latter two conditions were induced by 500 ppm CO inhalation for 5-6 weeks. Using an anesthetized open-chest preparation, these rats were compared with normal rats. With CO + PC + CM present, resting cardiac index, stroke index, stroke work, and minute work were elevated (heart rate also in the conscious state), while left ventricle end-diastolic pressure (LVDP) was normal. With PC + CM after CO washout, cardiac index and stroke index returned to normal at normal LVDP. Minute work, peripheral resistance, heart rate, and blood pressure, however, remained above normal. With CM alone, minute work, +dP/dtmax, +dF/dtmax, peripheral resistance, blood pressure, and LVDP declined from the condition with PC + CM. Although most cardiovascular parameters increased in the three conditions above with acutely increased LVDP, only with CM alone was performance augmentation normal. The results (i) reveal several characteristics of the hemodynamic response to chronic carboxyhemoglobinemia, (ii) suggest that the transient hypertension attending CO elimination in the presence of PC results from rapid reversal of peripheral vasodilatation, (iii) demonstrate decreased cardiac functional reserve with CO and (or) polycythemia upon preload challenge, and (iv) provide evidence for the benign nature of CO-induced cardiomegaly alone, on heart function. PMID- 2966663 TI - Interactions of cholecystokinin, beta-endorphin, and their antagonists on passive avoidance behavior in rats. AB - The effects of cholecystokinin octapeptide (CCK-8), cholecystokinin tetrapeptide amide (CCK-4), beta-endorphin, proglumide, and naloxone on passive avoidance behavior were studied in rats. Intracerebroventricular (i.c.v.) injection of beta endorphin (1-10 micrograms) had no significant influence on the latency of the avoidance response in intact rats. Also, beta-endorphin (0.05-5 micrograms, i.c.v.) did not affect the response in rats treated with electroconvulsive shock (ECS). The preventive effect of CCK-8 (0.1-1.0 micrograms, i.c.v.) on ECS-induced amnesia was partly antagonized by beta-endorphin (0.05-10 micrograms, i.c.v.). Intraperitoneal (i.p.) injection of naloxone (1-10 mg/kg) could not prevent ECS induced amnesia, but continuous subcutaneous infusion of this drug (2 mg/day, 7 days) completely abolished the amnesia. Naloxone (1 and 10 mg/kg, i.p.) also partly antagonized amnesia induced by proglumide (1 and 10 micrograms, i.c.v.) and prevented it when induced by CCK-4 (5 and 10 micrograms, i.c.v.). The results indicate the facilitating action of naloxone and the inhibitory effect of beta endorphin on memory, suggesting that the endogenous opiate systems are involved in some way in the memory processes. PMID- 2966664 TI - Ethanol-induced cardiac hypertrophy: effects of peripheral sympathectomy. AB - The development of cardiac hypertrophy was examined in rats that had undergone sympathectomy with 6-hydroxydopamine. After 4 days, the rats were given severely intoxicating doses of ethanol or isocalorically paired quantities of maltose dextrin by intubation at 8-h intervals up to 48 h. The ethanol and sugar intubations were applied in a nutritionally adequate, liquid diet mix. The extent of the peripheral sympathectomy was evident from the absence of detectable quantities of noradrenaline in hearts of animals injected with the neurotoxin and in the reduced levels of excreted noradrenaline. The adrenal medullary catecholamine contents of sympathectomized rats were unchanged in the absence of ethanol; there were reduced quantities of adrenaline in the medullae of rats given ethanol. The adrenal glands of rats given ethanol were larger than those from control animals. Urine samples from sympathectomized and control rats, given ethanol, displayed equivalent increases in excreted adrenaline and noradrenaline. Increases in relative cardiac weight were evident in hearts from sympathectomized animals after 4 days of sympathectomy, and this change reached significance in the hearts from 6-hydroxydopamine-treated rats after a further 2 days on the control diet. Hearts from animals exposed to ethanol showed a marked, rapid development of cardiomegaly; after 24 h there was an increased mass of some 17%, which was sustained over the remaining 24-h period. The proportion of cardiac protein did not differ in the large hearts from ethanol-treated animals and those from their controls, hence myocardial oedema could not account for the increase in weight.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966665 TI - The action of quinolinate in the rat spinal cord in vitro. AB - The responses of dorsal horn neurones to the excitatory amino acids quisqualate, kainate, N-methyl-D-aspartate (NMDA), and quinolinate have been examined in an in vitro preparation of the rat spinal cord. The antagonism of these responses by iontophoretically applied D-(-)-2-amino-5-phosphonovalerate (DAPV), kynurenate, and acridinate was tested, and the results were compared with data obtained from the spinal cord in vivo. The pattern of antagonism was similar in both preparations, although the potencies of agonists and antagonists were found to be significantly greater in vitro. The antagonism of amino acid induced firing of neurones was also recorded during the application of DAPV and kynurenate in the bathing medium. Dose-response curves and IC50 values were determined for these antagonists against all four agonists. The responses to quinolinate were antagonized differently from those to NMDA, quisqualate, or kainate, suggesting that quinolinate does not act specifically through the NMDA receptor as it does in other regions, nor does it appear to act via two or more of the three archetypal amino acid receptors. These findings suggest that a fourth amino acid receptor responsible for quinolinate's action in the spinal cord may exist. PMID- 2966666 TI - Cyclosporin A metabolites suppress T-cell proliferation by concanavalin A and in a mixed lymphocyte reaction. AB - To ascertain if cyclosporine metabolites (CMs) have immunosuppressive activity, bile, whole blood and urine taken from patients after cholecystectomy and from a liver transplant recipient on cyclosporin A (CsA) were assayed to determine their effect on T-cell proliferation induced by concanavalin A (Con A) and in a two-way mixed-lymphocyte response. Bile and whole blood from the liver transplant patient completely suppressed Con A proliferation and the mixed lymphocyte response (MLR) at dilutions at which normal bile and whole blood have no suppressive activity, but no such activity was noted from the urine. The CMs were separated into six peaks (fractions) by high-pressure liquid chromatography (HPLC). Metabolites were identified by internal standards and HPLC/mass spectrophotometric analysis. Cyclosporine metabolite fractions 2 and 3 from bile and blood had immunosuppressive activity similar to parent CsA whereas fractions 5 and 6 demonstrated less but substantial immunosuppressive activity. Fractions 2, 3 and 5 demonstrated an ability to inhibit two-way MLR similar to parent CsA; the other metabolite fractions were able to inhibit the MLR but to a lesser extent. These results demonstrate that a number of CMs have immunosuppressive effects similar to those of the parent compound, and this may account for the lack of correlation between whole blood CsA levels and immunosuppressive activity. PMID- 2966667 TI - Clinical trial of high-dose oral medroxyprogesterone acetate in the treatment of metastatic breast cancer and review of the literature. AB - Recent studies have suggested that there are benefits from the use of high-dose parenteral medroxyprogesterone acetate (MPA) in the treatment of metastatic breast cancer. The present study was designed to assess the efficacy and toxicity of high-dose oral MPA in women with clinical parameters suggesting potentially hormonally sensitive metastatic breast cancer. The first 28 patients received 800 mg/day, and 11 of them had received no previous hormone (NPH) therapy. The response rate (complete plus partial) was 63% for those receiving NPH and 12% for those receiving previous hormone (PH) therapy. Toxicity was significant at these doses, especially for women treated for more than 5 weeks. Toxic effects included excessive weight gain, Cushingoid facies, worsening of diabetes mellitus, and other stigmata suggestive of hypercorticism. Nineteen other patients were treated at 400 mg/day with a 60% response rate for 10 NPH patients and 44% for patients with PH treatment. Toxicity was less severe in these patients. The median time to treatment failure was 23 weeks, and to survival, 119 weeks for all treated patients. Moderately high (400 mg/d) and higher dose (800 mg/d) oral MPA are capable of inducing reasonable response rates in patients with NPH treatment. The toxicity of these regimens was significant--profound weight gain was dose limiting in some patients. While effective, high-dose oral MPA is unlikely to supplant tamoxifen as first-line therapy in metastatic breast cancer. PMID- 2966668 TI - Effects of antiestrogen and progestin on immune functions in breast cancer patients. AB - Several immunologic variables were evaluated in 14 patients with untreated primary breast cancer and 20 postmastectomized patients undergoing tamoxifen (TAM) or high-dose medroxyprogesterone acetate (MPA) treatment. Immunologic evaluation in the peripheral blood included lymphocyte count, definition of T lymphocyte subsets by monoclonal antibodies (OKT3, OKT11, OKT4, and OKT8), and lymphocyte blastogenic response to phytohemagglutinin (PHA) and Concanavalin A (Con A). Moreover, the in vitro effect of TAM and MPA on the blastogenic response of peripheral lymphocytes from normal female subjects was tested. Primary breast cancer patients did not differ from controls in any of the variables tested. Similarly, the immunologic variables of the group treated with TAM were normal, with the exception of a slight reduction of the OKT4+/OKT8+ ratio. In MPA-treated patients, a reduction of the percentage of OKT4+ cells and a decrease of the OKT4+/OKT8+ ratio were observed. Moreover, response to PHA was reduced sharply. However, the addition of interleukin-2 (IL-2) to the culture medium restored PHA response. Likewise, the in vitro addition of MPA to peripheral blood lymphocytes from normal female subjects resulted in a sharp dose-dependent depression of PHA response while TAM was ineffective completely. The inhibitory effect of MPA was not evident when IL-2 was added simultaneously to the culture medium. These results show that the administration of high-dose MPA may alter immunocompetence as defined by T-lymphocyte subsets and response to mitogens. The latter effect may be related to a diminished production of IL-2. In contrast, TAM does not appear to have a significant immunodepressant action either in vitro or in vivo. PMID- 2966669 TI - Down's syndrome and acute myelofibrosis. Time study of DNA content during the progression to leukemia. AB - Flow cytometry (FCM) for the determination of cellular DNA content was performed on multiple bone marrow biopsy specimens from a 3-year-old boy with Down's syndrome and myelofibrosis. A rapidly fatal acute nonlymphocytic leukemia developed within 3 months after initial bone marrow evaluation. The clinical and morphologic changes corresponded to the development of aneuploidy as determined by FCM and cytogenetic analysis. These findings support the clinical observations of the premalignant potential of myelofibrosis in Down's syndrome. PMID- 2966670 TI - Significance of bacterial flora in abdominal irradiation-induced inhibition of lung metastases. AB - We have previously reported that abdominal irradiation prior to i.v. injection of syngeneic tumor cells reduced metastases in lung. Our report described an investigation of the significance of intestinal organisms in the radiation effect. We found that eliminating intestinal organisms with antibiotics totally abolished the radiation effect. Monoassociation of germ-free mice revealed that the radiation effect was observable only for Enterobacter cloacae, never for Streptococcus faecium, Bifidobacterium adlesentis, or Escherichia coli. After abdominal irradiation of regular mice, E. cloacae multiplied in cecal contents, adhered to mucous membranes, invaded the cecal wall, and translocated to mesenteric lymph nodes. Intravenous administration of E. cloacae in place of abdominal irradiation inhibited metastases. E. cloacae-monoassociated mice developed fewer metastases than germ-free mice, and the reduction was further enhanced by abdominal irradiation. We concluded that abdominal irradiation caused the invasion of E. cloacae from the mucous membrane of the intestine and inhibited formation of lung metastases. PMID- 2966672 TI - New chemotherapeutic drug sensitivity assay for colon carcinomas in monolayer culture. AB - Ten previously untreated colon carcinomas were tested for chemotherapeutic drug sensitivity in primary monolayer culture. Colon carcinomas were partly digested to groups of epithelial cells which plated with a mean efficiency of 42 +/- 9% (SE) on a collagen I-bovine serum albumin substrate in serum-free medium, producing patches of tightly adherent epithelial cells. The cultured cells were judged epithelial by the presence of cytokeratins, an epithelial cell surface epitope, junctional complexes, and brush borders. Each carcinoma was plated in 40 to 60 Petri dishes (35 mm), yielding a mean of 28 +/- 8 (SE) colonies per dish (6832 +/- 1952 cells). Drugs tested in duplicate plates were mitomycin C, cisplatin, streptozotocin, and 5-fluorouracil at 0.1, 1, 10, and 100 micrograms/ml, and at 0.1, 1, and 2x the peak tolerated drug concentration in serum. Twenty-four h after plating, any nonadherent cells were removed, and the adherent tumor cells were continuously exposed to the drugs for 3 days. Each drug induced colony lysis in a dose-dependent manner in responsive tumors. Drug resistant, cycling cells were identified by [3H]thymidine incorporation in colonies which were not lysed by drug treatment. Each of the ten carcinomas exhibited inherent resistance to one or more chemotherapy drugs within the concentration ranges clinically achievable. PMID- 2966671 TI - Schedule-dependent therapeutic gain from the combination of fractionated irradiation and cis-diamminedichloroplatinum (II) in C3H/Km mouse model systems. AB - The interaction of cis-diamminedichloroplatinum(II) (c-DDP) with daily fractionated radiotherapy was studied in the SCCVII tumor, the duodenum, and the lungs of C3H/Km mice. The experimental end points were the time required for treated tumors to reach 3 times their treatment size, the survival of stem cells in the duodenal crypts, and the breathing rate measured early (19-23 weeks) and late (41-46 weeks) after treatment. In the 8 treatment schedules that were evaluated, radiation was delivered in 5 daily doses of 2-7 Gy, for total doses of 10-35 Gy; and c-DDP was administered either daily (2.4 or 1.6 mg/kg/day) or as a single bolus (8 or 12 mg/kg). Schedule 2, in which 2.4 mg/kg c-DDP was administered immediately before X-ray on 5 consecutive days produced the highest degree of enhancement of radiation effect (expressed as dose-effect factor); and the next greatest enhancement was produced by 12 mg/kg c-DDP administered 24 h before the start of fractionated daily radiotherapy. Those schedules also caused some enhancement in the normal tissues, but the dose-effect factors for those tissues were lower than for the tumor, which was reflected in the finding of maximal therapeutic gain factors for those same schedules. There was little or no enhancement nor were the therapeutic gain factors significantly greater than 1.00 when the 2 modalities were administered more than 24 h apart. Thus, for both normal tissue toxicity and antitumor effect there is striking schedule dependence with respect to both sequence and timing of these 2 modalities. This is of major relevance in clinical treatment planning. PMID- 2966673 TI - [The effect of a single administration of captopril on portal flow as measured by the Doppler method]. PMID- 2966674 TI - Secretory morphology of the intermediate lobe of the rat pituitary incubated in vitro. AB - The ultrastructure of the incubated intermediate lobe of the rat pituitary and the morphological effect of isoproterenol stimulation on its cells were studied under in vitro conditions. The general structure of isolated neurointermediate lobes maintained for 2-3 h in vitro was well preserved, and the presence of intact nerve terminals establishing synaptic contacts with the glandular cells of the intermediate lobe was confirmed. Removal of the intermediate lobe from central inhibition leads to increased hormonal secretion, which was reflected by large Golgi areas and the appearance of secretory images. However, no obvious degranulation or peripheral migration of the secretory granules after 2-3 h in vitro was seen. The secretory granules varied in electron density; totally electron-lucent granules were regularly observed and exocytotic phenomena were shown. In addition, more extensive invaginations suggesting secretion by compound exocytosis were seen. A three-fold increase in the beta-endorphin secretion during a 4-min stimulation with 10(-6) M isoproterenol did not induce any morphometrically detectable changes in the incubated cells. This indicates that only a minor fraction of the total granule content is mobilized during an acute increase in secretory activity. PMID- 2966676 TI - The balloon on a wire device: a new ultra-low-profile coronary angioplasty system/concept. AB - A new ultra-low-profile coronary angioplasty device, a "balloon probe," is presented. This device consists of a balloon on a coronary guidewire and is the lowest profile dilatation system presently available. The balloon material is polyethylene terephthalate, a new polymer with high inflation pressure limits and very low compliance. This report discusses this device and its usage in coronary artery disease. PMID- 2966675 TI - Distribution of atrial natriuretic polypeptide (ANP)-containing cells in the rat heart and pulmonary vein. Immunohistochemical study and radioimmunoassay. AB - The distribution of atrial natriuretic polypeptide (ANP) was immunohistochemically surveyed in the rat heart and lung using an antiserum raised against alpha-human ANP. The ANP-immunoreactive cells were seen to be distributed in the atrial walls and proximal portions of the pulmonary vein and venae cavae, but were absent from the aorta, pulmonary arteries, trachea, bronchus, and alveolar cells. The immunoreactive cells were present in a narrow region just beneath the endothelium of the pulmonary vein and vena cavae, and, ultrastructurally and immunocytochemically, were seen to be striated muscle cells with ANP-containing specific granules similar to those seen in atrial cardiocytes. A radioimmunoassay for ANP revealed a content of 604 +/- 51 pg/mg wet weight in the pulmonary vein, and 3343 +/- 1620 pg/mg wet weight in the venae cavae. In addition to the atrial wall, the proximal portion of both the pulmonary vein and venae cavae are suggested to be constituents of an ANP-producing organ. PMID- 2966677 TI - Coronary angioplasty at the time of initial diagnostic coronary angiography in patients with unstable angina. AB - Of the last 200 consecutive patients undergoing PTCA procedures at our institution, 29 (15%) had unstable angina; and angioplasty was performed at the time of diagnostic coronary arteriography. There were 26 males and three females with an age range of 31-82 (mean 57) years. Factors favoring PTCA at the time of initial coronary arteriography included clinical indications for revascularization, appropriate anatomy based on high-quality fluoroscopy, and availability of emergency surgery if required. Of 34 coronary lesions in 29 patients, 19 involved the anterior branch of the left anterior descending coronary artery (LAD), eight the circumflex branch (Cx); and seven the right coronary artery (RCA). Five patients had two vessels dilated (one LAD + RCA, two LAD + Cx, and two RCA + Cx). Of the coronary artery lesions, 19 were concentric, seven were eccentric, 20 were single and discrete, six were long or multiple in the same vessel; eight vessels were totally occluded, and in nine patients there was good collateral circulation. Twenty-nine (85%) arteries were successfully dilated. Of the unsuccessful cases, one was from failure to cross a totally occluded lesion, and three residual lesions and/or postdilatation pressure gradients remained significant. One patient required emergency aortocoronary bypass surgery because of total occlusion of the LAD immediately post-PTCA. There were no postprocedural myocardial infarcts or deaths. It is concluded that, in selected patients with unstable angina, PTCA can be performed successfully and with low risk at the time of initial diagnostic coronary arteriography. This approach offers certain clinical financial advantages. PMID- 2966678 TI - Blood velocity measurements during selective coronary angiography before and after percutaneous transluminal coronary angioplasty. AB - The velocity of blood flowing down a coronary artery may provide an index of myocardial perfusion, independent of the need for measuring the amount of myocardium supplied by a vessel. The velocity of the leading edge of contrast material was therefore measured before and after percutaneous transluminal coronary angioplasty in 15 patients utilizing digitized images from routine coronary angiography. The velocity (mean +/- SD) before percutaneous transluminal coronary angioplasty in the 15 patients was 11.9 +/- 6.0 cm/s, increasing to 21.7 +/- 8.7 cm/s after (P less than 0.01). There was a correlation between the percent change in velocity and the change in percent stenosis before and after percutaneous transluminal coronary angioplasty (r = 0.65; P less than 0.001). The mean absolute interobserver and intraobserver variabilities for the velocity measurements were 2.1 and 1.8 cm/s, respectively. Measurement of coronary flow velocity from data obtained at the time of routine coronary angiography is an easily performed reproducible technique, which may be used to assess the results of an intervention such as percutaneous transluminal coronary angioplasty. PMID- 2966679 TI - Application of a tapered teflon dilator to percutaneous transluminal coronary angioplasty. AB - A tapered teflon dilator has been applied to 12 cases of percutaneous transluminal coronary angioplasty (PTCA). The technique was first used to predilate stenoses that could not initially be crossed with low profile over the wire PTCA balloons. In the first five cases, three angioplasties that would have been failures were converted to successes by this method. The dilator was used in place of a balloon catheter as an initial step to assess the ability of a guidewire to cross a lesion to reduce cost in five additional cases. Modification of an existing family of peripheral vascular dilators is described. The method of application of this technique to PTCA is outlined. PMID- 2966680 TI - Successful combined percutaneous aortic balloon valvuloplasty and coronary angioplasty in two critically ill patients. AB - This report describes combined aortic balloon valvuloplasty and coronary angioplasty in two critically ill patients with aortic stenosis and coronary artery disease. PMID- 2966681 TI - Ectopic expression of a minor Drosophila opsin in the major photoreceptor cell class: distinguishing the role of primary receptor and cellular context. AB - We have used P-element-mediated transformation to introduce the cloned Rh1 rhodopsin gene into the germ line of Drosophila and fully rescue the visual phenotype of mutant ninaE flies. A transcriptional fusion between the ninaE promoter and the structural gene for a minor opsin (Rh2) that is not normally expressed in the R1-R6 photoreceptor cells was used to demonstrate that Rh2 rhodopsin can photoactivate the R1-R6 transduction cascade, but with different spectral sensitivity. In addition, we show that two mutants that specifically affect the R1-R6 cells, ninaA and rdgB, do not directly affect expression of the ninaE gene. PMID- 2966682 TI - Envelope glycoprotein of HIV induces interference and cytolysis resistance in CD4+ cells: mechanism for persistence in AIDS. AB - Masking of host cell receptors following retroviral infection is the basis for the phenomenon of virus interference. Amphotropic retrovirus vectors were used to express the HIV envelope glycoprotein in a human CD4+ cell line. Envelope expression is accompanied by a reduction in the level of surface CD4 receptor molecules and correlates with the presence of intracellular envelope-CD4 receptor complexes. Cells expressing the HIV envelope acquire a cytolysis-resistant phenotype such that infection with HIV leads to a non-cytopathic persistent virus infection. Furthermore, phorbol ester-mediated stimulation of viral replication in persistently infected cells results in renewed cytolytic effects which, due to the absence of CD4 in the cell population, are absolutely independent of syncytium formation. This study elucidates the mechanism by which viral persistence is initiated and maintained in the course of AIDS. PMID- 2966683 TI - Control of helper-T-cell proliferation by recognition of Ia and Mac-1 antigens on phagocytic cells of the thymic reticulum. AB - Phagocytic cells of the thymic reticulum (P-TR) have been previously described as being Ia-positive, Mac-1-positive accessory cells which pursue a close relationship with thymocytes. They form rosettes with thymocytes, and these rosettes are inhibited by antibody directed against the complement receptor type 3 CR3 (anti-Mac-1). P-TR induce the proliferation of syngeneic thymocytes. In the present paper, we show that thymocytes enriched in mature medullary type are induced to proliferate in coculture with syngeneic P-TR, while the cortical type does not. After 5 days of culture, 85% of the thymocytes are of helper L3T4+Lyt-2 phenotype. As previously shown by others for syngeneic reactions, antibodies directed against related class II antigens (anti-I-A and anti-I-E) block this helper-T-cell syngeneic proliferation. A new finding is the blockage of helper-T cell proliferation by anti-Mac-1 as well as with anti-LFA-1 antibodies, showing that accessory molecules may be as important as specific recognition of class II antigen molecules in the control of thymocyte proliferation and hence in thymocyte selection. Mac-1, like LFA-1, belongs to a novel family of differentiation antigens involved in cell interactions. The blockage of cell recognition and interaction between P-TR and thymocytes by either anti-Ia or anti Mac-1 during the early induction phase of the syngeneic response leads to its inhibition. We demonstrate that P-TR/thymocyte interaction stimulates the enhanced expression of IL-2 receptors on thymocytes, a step which is necessary for helper-T-cell proliferation. The mechanism of syngeneic proliferation inhibition by anti-Ia, anti-Mac-1, and LFA-1 antibodies may be the prevention of IL-2 receptor expression on thymocytes, and/or the inhibition of IL-2 secretion. Although this is an in vitro model, which may not totally reflect in situ situation, our results indicate that thymic accessory cells may participate in a positive selection process which leads to helper-T-cell proliferation. PMID- 2966684 TI - Replacement of the promoter of the yeast plasma membrane ATPase gene by a galactose-dependent promoter and its physiological consequences. AB - In order to probe the physiological role of the yeast plasma membrane ATPase we have replaced the constitutive promoter of its gene by a galactose-dependent promoter. The resulting cells stop growing on glucose medium when the preformed ATPase is diluted to 20% of normal. There is a correlation between ATPase activity and both proton efflux from the cells and amino acid transport. A large proportion of growth-arrested cells appear enlarged and with several buds containing nuclei. PMID- 2966685 TI - [What every pediatrician should know about the benefits and social services for handicapped children and their families. II]. PMID- 2966686 TI - The effect of antidepressant drugs on 24-hour rhythms of tryptophan metabolism in the rat. AB - The effect of three antidepressant drugs, imipramine hydrochloride, clomipramine hydrochloride and zimelidine dihydrochloride on plasma free and total TRY concentrations has been examined. The drugs were administered to male Wistar rats in the drinking water at a concentration of 200 micrograms ml-1 for up to 15 days. The effects of TRY concentration were complex and, after 2 days, inconsistent. After 14 days, the overall effect of imipramine was to lower free TRY levels and abolish 24-hr variation in free TRY concentration. Zimelidine and clomipramine tended to increase free TRY, particularly at 1300h so that the overall effect was to delay the peak in plasma-free TRY until later in the day. The possible significance of these findings to the mechanism of action of antidepressant drugs in clinical practice is discussed. PMID- 2966687 TI - On the role of energy metabolism in Neurospora circadian clock function. AB - Neurospora crassa (bdA) mycelia were kept in liquid culture. Without rhythmic conidiation the levels of adenine nucleotides undergo circadian changes in constant darkness. Maxima occur 12-17 hr and 33-35 hr after initiation of the rhythm, i.e., at CT 0-6 hr. Pulses of metabolic inhibitors such as vanadate (Na3Vo4), molybdate (Na2MoO4 : 2 H2O), N-ethylmaleimide (NEM), azide (NaN3), cyanide (NaCN) and oligomycin phase shift the circadian conidiation rhythm of Neurospora crassa. Maximal advance phase shifts are observed at about CT 6 with all inhibitors. Pulses of N,N'dicyclohexylcarbodiimide (DCCD) and light phase shift the conidiation rhythm following a phase response curve different from those of the other agents (maximal advance at about CT 18-24). The phase shifts with DCCD and light are significantly larger in the wild type compared to the mitochrondrial mutant poky. Such differences are not found in PRCs of the protein synthesis inhibitor cycloheximide. [31P] NMR spectra of wild type Neurospora crassa and the clock mutants frq 1 and frq 7 which differ in their circadian period lengths did not reveal differences in the concentrations of adenine nucleotides, pyridine nucleotides or sugar phosphates. Starvation causes drastic changes of the levels of adenine nucleotides, phosphate and mobile polyphosphate without effecting phase or period length of the circadian rhythm. PMID- 2966688 TI - Lack of variation over 24-hours in response to stimulation of 5-HT1 receptors in the mouse brain. AB - Two behavioural measures of activity at 5HT1 receptors in mice: RU 24969-induced hyperactivity and 8OH-DPAT hypothermia, were measured at 3-hr intervals throughout the 12-12 L-D cycle. Neither parameter showed significant circadian variation. PMID- 2966689 TI - Natural anticoagulant proteins in adult nephrotic syndrome. PMID- 2966690 TI - Heart with circulatory failure secretes and processes atrial natriuretic peptide in a manner different from normal heart. AB - The role of catecholamine in atrial natriuretic peptide (ANP) secretion and its secretory mechanism in normal humans is not well defined; therefore, we studied the relationship among ANP, catecholamine, and atrial pressures in 25 patients without cardiovascular disease and in 35 patients with chronic congestive heart failure (CHF, 20 in mitral valve disease and 15 in dilated cardiomyopathy). In patients without cardiovascular disease, right atrial pressure at rest showed a positive correlation (r = 0.80, p less than 0.001) with ANP concentration, whereas left atrial pressure did not. The relation narrowed (r = 0.82) when the bicycle ergometer exercise in the supine position was conducted. Neither adrenalin nor noradrenalin significantly correlated with ANP concentration. In patients with mitral valve disease and dilated cardiomyopathy, the significant relations (r = 0.56 p less than 0.001, r = 0.85 p less than 0.001, respectively) between left atrial pressures and ANP concentrations at rest were observed, and following exercise, induced more significant relations. Right atrial pressures did not correlate positively with ANP concentrations. The increments of ANP concentrations induced by exercise load were markedly reduced compared with those of patients without cardiovascular disease. Although concentrations of both noradrenalin and adrenalin in patients with mitral valve disease and dilated cardiomyopathy at rest were much higher than those without cardiovascular disease, only noradrenalin had a highly positive correlation with ANP concentrations (r = 0.88 p less than 0.001, r = 0.78 p less than 0.001, respectively). Furthermore, the circulating ANP molecular weight forms in all patients studied were analyzed by gel chromatography coupled with radioimmunoassay.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966691 TI - Steroid synthesis inhibition by ketoconazole: sites of action. AB - Ketoconazole is an antifungal agent that, in high doses, inhibits testicular and adrenal steroid synthesis. The ability of ketoconazole to block steroid synthesis has prompted us to use it in the treatment of advanced prostatic carcinoma. This study was designed to determine the site of steroid synthetic blockade that was induced by ketoconazole. Twelve patients with metastatic prostate carcinoma on long term high dose ketoconazole therapy were compared with 12 control volunteers. Values of serum progesterone, 17-hydroxyprogesterone, androstenedione, dehydroepiandrosterone sulphate, testosterone, and cortisol were measured in a baseline state and after Cosyntropin and human chorionic gonadotropin stimulation. Baseline data showed that serum levels of testosterone, androstenedione, and dehydroepiandrosterone sulphate were lower and that plasma progesterone, luteinizing hormone, and adrenocorticotropin were higher in the ketoconazole group. With Cosyntropin, plasma cortisol, androstenedione, and dehydroepiandrosterone sulphate increased only in the control group. With human chorionic gonadotropin, testosterone increased only in the control group. Basal 17-hydroxyprogesterone and progesterone rose after Cosyntropin only in the ketoconazole group. Following human chorionic gonadotropin, progesterone rose in the ketoconazole group but not in the control group. These results suggest that ketoconazole is a potent inhibitor of steroid synthesis. The major site of action appears to be in the inhibition of 17-20 desmolase. A moderate blockade of 17 hydroxylase may be present. There is a marked inhibition of 21- and/or 11 hydroxylase. The ability of ketoconazole to inhibit steroid synthesis should have therapeutic potential in the treatment of steroid dependent disease. Frequent high dose ketoconazole therapy can inhibit adrenal steroid synthesis, which can be important for patients undergoing stressful situations. PMID- 2966692 TI - Increase in specific binding of insulin-like growth factor (IGF) II to type 1 IGF receptors on erythrocytes of hypopituitary children receiving growth hormone therapy. AB - Specific receptor binding for insulin-like growth factors (IGFs) is measurable in young erythrocytes. Cells of similar age, Fraction A, can be reproducibly obtained by dextran gradient centrifugation from 5-10 ml of blood. We now report IGF-II specific binding to Fraction A erythrocytes from normal children and children with growth hormone deficiency. Normal controls (Group 1) were 5 male volunteers (14.7 +/- .6 years, mean +/- SEM) and 10 children with constitutional short stature (11.4 +/- 1.6 years) who had normal 6-hour daytime growth hormone profiles and plasma IGF-I values. Twelve growth hormone deficient children (Group 2), aged 13.7 +/- 1.1 years, had samples taken after 2 months without growth hormone therapy and again following 2 months with growth hormone (0.1 U/kg 3 times per week) therapy. The percent of total erythrocytes in Fraction A did not differ in the two groups of children. Group 1 had IGF-II specific binding of 10.2 +/- 0.6% (per 3 X 10(9) cells). IGF-II specific binding was less in Group 2 at 6.6 +/- 0.8% (p less than 0.002). With growth hormone therapy, IGF-II specific binding increased to 10.4 +/- 1.0% (p less than 0.02), a value not different from that seen in Group 1. Corresponding plasma IGF-II and IGF-I values showed a positive correlation with IGF-II specific binding (r = 0.54 and r = 0.56 respectively, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966693 TI - The pharmacokinetics of etodolac in serum and synovial fluid of patients with arthritis. AB - The pharmacokinetics of etodolac have been evaluated in five patients with arthritis given 200 mg etodolac, twice daily, at 12-hour intervals, for 7 days. Albumin and total protein concentrations were markedly lower in synovial fluid than in serum, and etodolac free fraction was significantly higher. Etodolac readily penetrated into the synovial fluid, and in the postdistributive phase the concentration of free etodolac (i.e., the drug responsible for pharmacologic activity) remained higher than that in serum at all times. No differences in the half-life of etodolac elimination were noted. PMID- 2966694 TI - Effects of aspirin, naloxone and placebo. AB - This study was designed to examine the effects of aspirin, naloxone and placebo treatment on serum beta-endorphin concentration and joint pain in patients with rheumatoid arthritis (RA). Ten patients with definite or classical RA were studied. All treatments were administered in a randomized sequence. On each study day, the following measurements were carried out at specified time intervals: serum beta-endorphin concentration, serum salicylate concentration and joint pain score on a visual analogue horizontal scale. We conclude that in patients with rheumatoid arthritis suffering from chronic joint pain, serum beta-endorphin does not appear to play a role in pain relief. PMID- 2966695 TI - Allergic contact dermatitis from dibrompropamidine cream. PMID- 2966696 TI - Allergic and irritant contact dermatitis from diethyl-beta-chloroethylamine. PMID- 2966697 TI - Contact dermatitis from Emulgin RO/40, an emulsifier in Hioxyl cream. PMID- 2966698 TI - Allergic contact dermatitis caused by an acetone-formaldehyde condensate. PMID- 2966699 TI - Occupational dermatitis from Mesonia chinensis. PMID- 2966701 TI - Sensitisation to dichlorobenzoyl chloride. PMID- 2966700 TI - Allergic contact dermatitis to laureth-4. PMID- 2966702 TI - Patch testing with dithranol and its degradation products. PMID- 2966703 TI - Acne and folliculitis due to diesel oil. PMID- 2966704 TI - Occupational dermatitis from gold plating. PMID- 2966705 TI - Identification of undetected corticosteroid allergy. PMID- 2966706 TI - Contact allergy to imidazole antimycotics. AB - Between 1977 and 1986, 9 patients with contact allergy to the active ingredient of imidazole antimycotics were found at the Department of Dermatology, University of Heidelberg. The number of positive reactions decreased in the following order: miconazole (6), clotrimazole (3), econazole (3), isconazole (3), and oxiconazole (1). When 5 patients were tested with a series of imidazoles in different concentrations and vehicles (petrolatum, ethyl methyl ketone, ethanol), petrolatum turned out to be the least effective one. The active ingredient at 1% in ethanol seems to be the most suitable choice for routine patch testing. Bifonazole may be the therapeutic alternative for patients sensitive to miconazole or clotrimazole, since no cross reactivity was observed. PMID- 2966707 TI - Occupational dermatosis among chrome platers. AB - 14 (38%) of 37 chrome platers in 17 chrome electroplating factories surveyed had occupational contact dermatitis, chrome ulcers, or both. 7 had chrome ulcers, 6 had contact dermatitis and 1 had both. Another 16 (43%) workers had scars suggestive of previous chrome ulcers. Mucosal irritation was present in 57% of the workers. The most common was throat irritation (49%) followed by nasal irritation (41%). Mucosal irritation was more common in hard chrome platers, while skin ulcers and dermatitis were more common in bright chrome platers. Nasal septum perforation was seen in 1 worker. Skin ulceration appeared to be a more specific sign for occupational dermatosis in chrome platers than dermatitis when the prevalence rates were compared to controls. Of the 7 workers with chrome ulcers, only 1 was allergic to chromate. Of the 6 workers with dermatitis, 2 were allergic to chromate and 1 to nickel. The worker with ulceration and dermatitis was negative to chromate and nickel. Irritant factors are therefore important in the aetiology of contact dermatitis in these chrome platers. PMID- 2966708 TI - Occupational allergic contact dermatitis in beauticians. AB - We patch tested 13 beauticians with hand dermatitis between 1982 and 1986. They were all young female novice beauticians or those in training. The onset of their allergic dermatitis was noticed within 1 month to 1 year of their starting this occupation. Definite positive reactions to products were seen from hair dyes (as is, open test) (6/12), cold permanent wave primary solutions (as is, open test) (7/13) and a shampoo (1% aq., closed test) (1/13). Positive reactions to allergens were seen with para-phenylenediamine (1% pet) (12/13), ammonium thioglycolate (5% aq., open test) (3/7), para-toluylenediamine (1% pet) (7/9), para-aminophenol (1% pet) (1/4), ortho-aminophenol (1% pet) (1/4), Quinoline yellow SS (0.5% pet) (1/4), nickel sulfate (2.5% pet) (1/12), cobalt sulfate (2.3% pet) (1/12), thimerosal (0.05% pet) (1/12) and procaine hydrochloride (1% pet) (1/12). Study of the prognosis showed that 5 out of 12 cases could continue their occupation, but 4 cases had persistent hand dermatitis despite protecting their hands from hair dyes with gloves, 7 cases quitted their jobs, and in 5 their hands healed while 2 cases continued to have atopic hand dermatitis. A personal or family history of atopy was frequent among the cases, so we recommend that those who have such a history should not become beauticians. PMID- 2966709 TI - Occupational allergic contact dermatitis from anaerobic acrylic sealants. AB - Anaerobic sealants are acrylates that polymerize in the absence of oxygen. They are used principally in the engineering and electronic industries. Their use can cause occupational dermatitis in the workers who handle them. In this work, we report our experience with 6 patients who presented with allergic contact dermatitis to these sealants. PMID- 2966710 TI - Evaluation of the sensitising potential of 4 polyamines present in technical triethylenetetramine using 2 animal species. AB - Contact sensitivity to 4 polyamines present in technical grade triethylenetetramine was investigated using the 'VAA mouse' assay and the guinea pig maximization test (GPMT). The criteria used to assess sensitivity in the GPMT classed all substances as having the same degree of sensitising potential. The mouse assay ranked N-(2-piperazin-1-ylethyl)ethylenediamine as having a significantly greater sensitising potential than the other compounds. PMID- 2966711 TI - Assessment of erythema in irritant contact dermatitis. Comparison between visual scoring and laser Doppler flowmetry. AB - Assessment of erythema in experimentally-induced irritant contact dermatitis has been performed visually and using the laser Doppler flowmeter (LDF). A close correlation was shown between the 2 methods (r = 0.9079, p less than 0.001), with the LDF producing mean blood flow values which were able to discriminate between the different visual scores. Of the 100 patch tests evaluated, 3 gave poor correlations between their visual and LDF readings, including 2 dithranol reactions and 1 sodium hydroxide response. Patch tests with no visible erythema had blood flow values similar to those of normal untreated skin. Although the LDF was an easy instrument to operate, it was not considered suitable for use in the routine patch test clinic, due mainly to the unacceptable length of time required to measure each patch test. PMID- 2966712 TI - Occupational contact dermatitis from vitamin K3 sodium bisulphite. PMID- 2966713 TI - Photodermatitis from anthraquinone. PMID- 2966714 TI - Allergic contact dermatitis from methyl heptine and methyl octine carbonates. PMID- 2966715 TI - Epidemic of occupational contact dermatitis from ranitidine. PMID- 2966716 TI - Epoxy resin dermatitis in a stained glass window maker. PMID- 2966717 TI - Reactions to cadmium. PMID- 2966718 TI - Occupational vitiligo. PMID- 2966719 TI - "Thin mint" cookie dermatitis. PMID- 2966720 TI - Contact urticaria to naphtha present in a solvent. PMID- 2966721 TI - An unfortunate complication of self-therapy for seborrheic dermatitis. AB - The death of a young boy who practiced self-therapy of seborrheic dermatitis of the scalp is reported. Practitioners should be aware that peer pressure in today's teenage society, coupled with misinterpretation of therapeutic protocols, may result in the application of unorthodox treatment regimens. PMID- 2966722 TI - Tretinoin-sunscreen mixture in the treatment of acne vulgaris. AB - In an open design study of fifty-four patients with grade I or grade II acne vulgaris, the combination of 0.1 percent tretinoin cream (Retin-A) and a sunscreen with sun protection factor 15 (Sundown) was evaluated. Overall study results of the forty-six patients who could be evaluated demonstrated a decrease in papule count of 25.9 percent, a decrease in closed comedones of 49.1 percent, and a decrease in open comedones of 36.3 percent by the end of the eight-week study period. These results indicate that the addition of a noncomedogenic sunscreen to tretinoin did not compromise its effectiveness and successfully prevented photoaccentuation reactions. PMID- 2966723 TI - Older volunteers and youths with disabilities team up to find jobs. PMID- 2966724 TI - Project pairs: a peer-assisted swimming program for the severely handicapped. PMID- 2966725 TI - Preparing the learning disabled adolescent for adulthood. PMID- 2966726 TI - [Ergonomic considerations on the daily working conditions in the dental office]. PMID- 2966727 TI - [The cancer patient and his place in oro-dental practice]. PMID- 2966728 TI - A new silicone-prosthesis in the modified Thiersch operation. AB - Experience with a new silicone prosthesis in the modified Thiersch operation for rectal procidentia in 16 extremely poor-risk patients is presented. The technique of implantation, structural details of the prosthesis, and the clinical results are described. The use of a new silicone prosthesis in the modified Thiersch procedure is a viable alternative in this group of patients. Surgical technique is a primary determining factor in preventing complications. PMID- 2966729 TI - Teicoplanin: a new glycopeptide antibiotic complex. AB - The chemistry, microbiology, pharmacokinetics, clinical efficacy, and adverse effect profile of teicoplanin are reviewed and, where appropriate, compared with vancomycin. Teicoplanin is a glycopeptide antibiotic with potent bactericidal activity against a wide variety of aerobic and anaerobic gram-positive bacteria. In contrast to the structurally related vancomycin, teicoplanin has a prolonged elimination half-life of approximately 60 hours and it may be safely administered by the intramuscular route. Adverse effects of teicoplanin include ototoxicity, nephrotoxicity, skin rash, eosinophilia, neutropenia, and transient elevation of serum aminotransferases. Teicoplanin may be beneficial as an alternative to vancomycin for patients with poor vascular access and in those requiring long term outpatient therapy. The role of teicoplanin in the treatment and prophylaxis of gram-positive infections will ultimately depend on its unfolding safety and efficacy profile. PMID- 2966730 TI - [Computer-assisted recording and processing of patch test data. A model]. AB - In the Department of Dermatology of the University of Graz (Austria) since 1985 an electronic data system is used for registration and evaluation of patch test data. The installed system (hardware and software), database and the possible methods for evaluation are presented. The necessity of global solution of the problem with possible access to complete database (patient data, product declaration and literature) is emphasized. PMID- 2966731 TI - [Skin and systemic disease caused by occupational contact with p-tert butylphenol. Case reports]. AB - Three patients who occupationally had been in contact with p-tert.-Butylphenol (ptB) developed vitiligo. Patch tests were negative to ptB. Antithyroid and antiparietal cell antibodies in all three patients indicate a possibly beginning systemic disorder such as already described in literature as the symptom triad of "vitiligo, hepatosplenomegalia and struma". PMID- 2966732 TI - Skin telangiectasia in workers of an aluminium processing plant. AB - Telangiectases were detected in 57 (46.7%) of 122 workers employed at the electrolysis department of the aluminium plant Lozovac/Sibenik, Yugoslavia. Of 116 workers employed in the smeltery of the same plant, telangiectases were observed in 19 persons (16.4%) and in a group of 121 public transportation workers from Sibenik 15 men (12.4%) displayed enlarged blood vessels. Statistical evaluation revealed the difference in number between workers showing telangiectases engaged in electrolysis and the other two samples to be significant. It may be assumed that telangiectases were caused by hydrogen fluoride and other fluorides. Further clinical parameters, as well as working conditions, are reviewed. PMID- 2966733 TI - Systemic contact-type dermatitis by derivatives of adamantane? AB - The possibility of system skin reactions caused by amantadine in patients sensitized for tromantadine is discussed. The practical question arises how to handle patients allergic tromantadine patients which may be helped by therapy with amantadine. PMID- 2966735 TI - The impact of computers on the writing process. PMID- 2966734 TI - Ultrastructural localization of erythrocyte cytoskeletal and integral membrane proteins in Plasmodium falciparum-infected erythrocytes. AB - The distributions of ankyrin, spectrin, band 3, and glycophorin A were examined in Plasmodium falciparum-infected erythrocytes by immunoelectron microscopy to determine whether movement of parasite proteins and membrane vesicles between the parasitophorous vacuole membrane and erythrocyte surface membrane involves internalization of host membrane skeleton proteins. Monospecific rabbit antisera to spectrin, band 3 and ankyrin and a mouse monoclonal antibody to glycophorin A reacted with these erythrocyte proteins in infected and uninfected human erythrocytes by immunoblotting. Cross-reacting malarial proteins were not detected. The rabbit sera also failed to immunoprecipitate [3H]isoleucine labeled malarial proteins from Triton X-100 and sodium dodecyl sulfate (SDS) extracts of infected erythrocytes. These three antibodies as well as the monoclonal antibody to glycophorin A bound to the membrane skeleton of infected and uninfected erythrocytes. The parasitophorous vacuole membrane was devoid of bound antibody, a result indicating that this membrane contains little, if any, of these host membrane proteins. With ring-, trophozoite- and schizont-infected erythrocytes, spectrin, band 3 and glycophorin A were absent from intracellular membranes including Maurer's clefts and other vesicles in the erythrocyte cytoplasm. In contrast, Maurer's clefts were specifically labeled by anti-ankyrin antibody. There was a slight, corresponding decrease in labeling of the membrane skeleton of infected erythrocytes. A second, morphologically distinct population of circular, vesicle-like membranes in the erythrocyte cytoplasm was not labeled with anti-ankyrin antibody. We conclude that membrane movement between the host erythrocyte surface membrane and parasitophorous vacuole membrane involves preferential sorting of ankyrin into a subpopulation of cytoplasmic membranes. PMID- 2966736 TI - Sustained release verapamil in renal hypertension. AB - In 14 patients with arterial hypertension secondary to chronic renal parenchymal disease and impaired renal function, 24-h ambulatory and casual blood pressure readings plasma, angiotensin II, aldosterone, arginine vasopressin and atrial natriuretic peptide, creatinine clearance, plasma lipids and lipoproteins, and body weight were determined after consecutive 3-week periods on placebo and sustained release verapamil 240 mg/day. Verapamil reduced the mean 24-h ambulatory blood pressure from 152/104 to 142/97 mm Hg. Blood pressure was significantly reduced during the daytime and the evening, but not at night. Casual blood pressure was also significantly reduced from 176/106 mm Hg to 154/96 mm Hg. No significant changes were found in the hormones, creatinine clearance, plasma lipids and lipoproteins, heart rate or body weight. The atrial natriuretic peptide level was significantly correlated with the calculated creatinine clearance (r = -0.797). Thus, sustained release verapamil 240 mg as a single daily dose had a moderate hypotensive effect in patients with chronic renal disease without inducing tachycardia, activation of the renin-angiotensin aldosterone system, or increasing body weight, and without altering renal function and plasma lipids and lipoproteins. The negative correlation between atrial natriuretic peptide and glomerular filtration rate supports the hypothesis that the extracellular volume increases during progression of renal disease. PMID- 2966737 TI - Long-term depletion of CD8+ T cells in vivo in the rat: no observed role for CD8+ (cytotoxic/suppressor) cells in the immunoregulation of experimental allergic encephalomyelitis. AB - A method is described for the long-term depletion of CD8+ T cells in the rat. Employing Lewis rats depleted of this cell type it is shown that such animals acquire, and subsequently recover from both passively and actively induced experimental allergic encephalomyelitis (EAE) in a comparable fashion to normal animals. Furthermore, after spontaneous recovery from the initial bout of disease, CD8+ T cell-depleted rats do not show any signs of relapse and, like normal rats following recovery from active EAE, are still resistant to further attempts at disease induction. Overall, these data do not support a role for the classic cytotoxic/suppressor (CD8+) T cell in the immunoregulation of EAE in the Lewis rat. PMID- 2966738 TI - A CD3- subset of CD4-8+ thymocytes: a rapidly cycling intermediate in the generation of CD4+8+ cells. AB - T lymphocytes with the surface phenotype CD4+8- and CD4-8+ are considered to be representative of functionally mature cells. We show here that adult murine thymus contains a subpopulation of CD4-8+ cells that differ from CD4-8+ cells found in the periphery in that they do not express the T cell receptor-associated CD3 molecular complex. Such CD3-4-8+ thymocytes are cortisone sensitive and rapidly cycling in situ. Furthermore, in contrast to mature T cells, most CD3-4 8+ thymocytes express low levels of CD5 and high levels of the B2A2 antigen. CD3 4-8+ thymocytes fail to respond to a variety of mitogenic stimuli in vitro but do give rise upon short-term culture to CD4+8+ cells. It is suggested that CD3-4-8+ thymocytes represent a transitional stage of thymus differentiation between the CD4-8- and CD4+8+ compartments. PMID- 2966740 TI - An adjunct trait of HEL/I-Ab-specific T helper cell is sensitivity to antigen specific immunosuppression. AB - The present study tests whether the specific inhibition of helper T (Th) cell (and T hybridomas) by suppressor T (Ts) cells is a phenotypic trait of Th cells correlating with their acquired specificity for antigen/major histocompatibility complex or a genotypic trait not related to selection of the T cell repertoire for antigen. To do this we took advantage of the fact that H-2d parental strains of mice commonly restrict recognition of chicken egg-white lysozyme to the L3 peptide (a.a. 105-129) and H-2b parental mice to the L2 peptide (a.a. 13-105). F1 hybrids of these strains display two subsets of lysozyme-reactive T cells, one for each parental phenotype. Using (B10 X B10.D2)F1 mice reconstituted with B10.D2 bone marrow, we were able to develop genetic H-2d T cell clones that could express an atypical specificity, that is L2/I-Ab. Clones of this type, like genetic H-2b, are also sensitive to the inhibiting effects of HEL-activated Ts cells. To overcome some of the drawbacks of using heterogeneous populations of T, B and accessory cells in our assays, we constructed T hybridomas from HEL-immune, chimeric lymph node T cell blasts which respond to a unique antigen/major histocompatibility complex with production of the lymphokine interleukin 2. Our results indicate that all HEL/I-Ab-specific T cells (helper and hybridomas) are inhibited by suppression regardless of the T cell's haplotype at the H-2 locus: H 2b (B10), H-2d (D2) or H-2b,d (BDF1). Furthermore, there is a strict correlation between the antigen and I-A specificity: I-Ab-restricted T cells recognize non-L3 determinants even though some are derived from H-2d mice. PMID- 2966739 TI - Signal transduction through CD4 receptors: stimulatory vs. inhibitory activity is regulated by CD4 proximity to the CD3/T cell receptor. AB - The binding of antibody to the CD4 molecule inhibits mobilization of cytoplasmic free calcium ([Ca2+]i) in response to CD3 cross-linking on resting T cells. Similarly, when CD3 and CD4 are independently and simultaneously cross-linked, calcium mobilization is inhibited when compared to that induced by cross-linking CD3 alone. In contrast, when anti-CD4 and anti-CD3 are cross-linked together, calcium mobilization is substantially higher than from CD3 cross-linking alone. A heteroconjugate consisting of covalently bound CD3 and CD4 monoclonal antibodies (mAb) retains the ability to mobilize [Ca2+]i in CD4 cells at protein concentrations approximately two orders of magnitude lower than the free CD3 mAb, and the activity of the heteroconjugate is inhibitable by free CD4 mAb. The CD3/CD4 heteroconjugate also shows significantly greater activity in stimulation of inositol phosphate IP1, IP2 and IP3 synthesis in T cells than the CD3 mAb alone, and again the activity is inhibited by free CD4 mAb. The activity of the CD3/CD4 heteroconjugate is not simply due to oligomerization, since CD3/CD3 or CD4/CD4 homoconjugates or homoconjugate mixtures did not show increased activity. Other heteroconjugates (CD3/CD5 and CD3/CD28) were not different than the CD3/CD3 homoconjugate in their ability to increase [Ca2+]i. Purified CD4 T cells that do not respond to CD3 mAb in solution do respond to the CD3/CD4 heteroconjugate in solution by proliferating in the presence of a CD28 mAb, with a significant fraction of CD4 cells entering the second cycle within the first three days of stimulation. The CD3/CD4 heteroconjugate co-modulates the CD3 and CD4 receptors, indicating that the heteroconjugate is not simply anchoring the T cell receptor to the T cell surface like anti-CD3 on a solid surface. These results suggest that CD4 plays an active role in signal transduction when brought into close physical proximity to the CD3/T cell receptor complex during major histocompatibility complex class II-restricted antigen presentation. PMID- 2966741 TI - T cell proliferation induced by anti-CD3 antibodies: requirement for a T-T cell interaction. AB - In the presence of interleukin 2 (IL2), soluble anti-CD3 monoclonal antibodies can stimulate highly purified normal T lymphocytes to proliferate. In these experiments HLADR+ T cells constituted 13 to 20% of the total cell population, and other HLADR+ cells, such as monocytes and B lymphocytes, constituted less than 1% of the population. When the HLADR+ T cells were removed from the total T cell population by cytofluorometric sorting, the residual HLADR- T cells failed to respond to soluble anti-CD3 plus IL2. When the separated HLADR+ T cells were recombined with the HLADR- T cells, a response was again found. This response was dependent on the dose of HLADR+ T cells added in the mixture. Irradiation individually of the HLADR+ and DR- T cells revealed that the proliferation in the cell mixture was predominantly, if not exclusively, by the HLADR- T cells. The ability of the HLADR+ T cells to provide a signal necessary to this proliferation was radioresistant. These data indicate that under the conditions of these experiments HLADR+ ("activated") T cells provide a signal necessary to the responsiveness of previously resting T cells. PMID- 2966742 TI - The effect of the thromboxane receptor antagonist BM 13.177 on experimentally induced coronary artery thrombosis in the pig. AB - We studied the effect of pretreatment with two doses of the thromboxane antagonist BM 13.177 and its solvent on the development of electrically induced coronary artery thrombosis in pigs. Results were compared with those obtained in animals pretreated with intravenously administered acetylsalicylate and its solvent. The effects of both compounds on the overall cardiovascular performance (heart rate, mean arterial blood pressure, cardiac output, systemic vascular resistance) were minimal. In the animals receiving solvent or acetylsalicylate the time to occlusive coronary thrombosis was 33 +/- 4 and 32 +/- 6 min, respectively. BM 13.177, in a dose of 5 mg.kg-1, did not modify the time to thrombotic occlusion (35 +/- 7 min), but in six of the eight animals that had received 10 mg.kg-1 BM 13.177, there was no occlusion within 120 min. In the acetylsalicylate-treated animals, collagen-induced platelet aggregation and plasma thromboxane B2 declined by 72 and 82%, respectively. The decreases were 46 and 20%, respectively, with the higher dose of BM 13.177. It is concluded that, in this porcine model of coronary artery thrombosis, the thromboxane antagonist BM 13.177 effectively suppressed formation of occlusive thrombi whereas acetylsalicylate was ineffective at a dose that lowered arterial thromboxane levels. PMID- 2966743 TI - Antihypertensive effects of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) in conscious dogs. AB - Low doses of 8-OH-DPAT (10 and 25 micrograms/kg) administered subcutaneously (s.c.) to renal hypertensive mongrel dogs caused decreases in systolic blood pressure which persisted for 3 h. Mild salivation and hyperventilation were observed with both doses. A short-lasting (less than 60 min) depressor response was seen with 100 micrograms/kg s.c. Prominent hyperventilation and salivation accompanied this response. A still higher dose (250 micrograms/kg s.c.) induced tremor, signs of anxiety and occasional vomiting in addition to the hyperventilation and salivation. Paradoxically, no cardiovascular activity was noted at this dose. PMID- 2966744 TI - Isoenzyme analysis of Echinostoma liei: comparison and hybridization with other African species. AB - Isoenzyme analysis was carried out on the laboratory strain of Echinostoma liei. The results were compared with those from a preliminary study on Echinostoma caproni, Echinostoma togoensis, and Echinostoma sp. (A. Voltz, J. Richard, B. Pesson, and J. Jourdane, 1986, Annales de Parasitologie Humaine et Comparee, 61, 617-623). Isoelectrofocusing showed characteristic phenotypes for phosphoglucomutase (PGM, EC 5.4.2.2) and glucosephosphate isomerase (GPI, EC 5.3.1.9). The four experimental strains were monomorphic. Their genotypes were defined. Isoenzyme analysis of F1-hybrids and their F2 descendants indicated the subunit structure of both isoenzymes and showed that they were encoded by independent genes. Finally, it also suggested that the four strains corresponded to variants of the same species. PMID- 2966745 TI - Role of endopeptidase-24.11 in the inactivation of atrial natriuretic peptide. AB - The circulating form of atrial natriuretic factor is a 28-residue peptide containing a 17-residue disulphide-linked ring. It has important actions on the kidney, largely on its haemodynamics, and at other sites including the adrenal cortex and CNS. It has a short half-life in vivo and is rapidly inactivated when incubated with kidney microvillar membranes. Of the battery of peptidases present in that membrane, only one, endopeptidase-24.11, is responsible for initiating the attack, and this commences with hydrolysis of the Cys7-Phe8 bond within the ring. Hydrolysis at this and other points has been shown to inactivate the peptide and this information has pointed the way to the synthesis of resistant analogues. PMID- 2966746 TI - Anoxic brain function: molecular mechanisms of metabolic depression. AB - An examination of the kinetic parameters of phosphofructokinase, pyruvate kinase and glycogen phosphorylase, and the cellular concentration of fructose 2,6 bisphosphate during anoxia in the turtle Pseudemys scripta showed that the total activity of glycogen phosphorylase, and the phosphofructokinase inhibition constants for citrate and ATP were decreased in anoxic turtle brain. These results suggest that the ability of turtle brain to survive extended periods of anoxia is the result of metabolic rate depression regulated, at the molecular level, by enzyme inactivation through anoxia-induced covalent modification. PMID- 2966747 TI - The conversion of phosphoserine residues to selenocysteine residues on an opal suppressor tRNA and casein. AB - This study has been undertaken in order to elucidate the mechanisms of incorporation of Se into glutathione peroxidase (GSHPx), in which selenocysteine corresponds to the opal termination codon UGA on the mRNA. We studied the above mechanisms using an opal suppressor tRNA, prepared from bovine liver, and casein as a model protein for the GSHPx apo-enzyme which might contain phosphoserine. The results showed that opal suppressor tRNA did not accept selenocysteine (lower than 0.1 mmol/mol) under the standard conditions. A trace amount of phosphoseryl tRNA was converted to selenocysteyl-tRNA by incubation with H2Se and some enzymes. Meanwhile, a number of phosphoserine residues in casein were converted to selenocysteine residues by incubation with H2Se and enzymes. These results suggest that opal suppressor tRNA plays a role in synthesizing GSHPx via co- and/or post-translational mechanisms. PMID- 2966748 TI - The cleavage of beta-chain in bovine fibrinogen DH fragment (95 kDa) leads to a significant increase in its anticlotting activity. AB - It is shown that in the presence of Ca2+ plasmin converts bovine fibrinogen fragment DH (95 kDa) into DLA fragment by the cleavage of its beta-chain Arg372 Thr373 bond. DLA fragment consists of two components (82 and 12 kDa) held together by non-covalent bonds and has 3.5-fold higher anticlotting activity than DH fragment. The DH to DLA fragment conversion leads to the destabilization of thermolabile domains of the latter without the loss of their compact structure. The results obtained show that the activation of DH fragment by the cleavage of its Arg372-Thr373 bond bears some resemblance to the general activation of proenzyme into enzyme. PMID- 2966749 TI - [Osteochondrosis of the lumbar spine]. PMID- 2966750 TI - Handicapped children: there is progress! PMID- 2966751 TI - Techniques for transporting the handicapped patient in the dental setting. PMID- 2966752 TI - Transesophageal real-time two-dimensional Doppler echography--a new method for the evaluation of azygos venous flow. PMID- 2966753 TI - More on tuberculous peritonitis and laparoscopy. PMID- 2966754 TI - [On the problem of the definition of ovarian pregnancy]. AB - We report on a case of true ovarian pregnancy, and discuss the anatomical criteria required for the diagnosis of this rare type of ectopic pregnancy. PMID- 2966756 TI - Karyological identification of two taxa of the Anopheles balabacensis complex from Burma. AB - Giemsa and Hoechst staining of neuroblast chromosomes were used to identify two strains of the Anopheles balabacensis complex from Burma. The laboratory colony of Kwan-ka-thaung (KKT) strain is shown to correspond to A. dirus A, while the laboratory colony of Taikkyi (TKK) strain corresponds to A. dirus C as defined by earlier studies on material from Thailand. PMID- 2966755 TI - Changes in DNA base sequence induced by gamma-ray mutagenesis of lambda phage and prophage. AB - Mutations in the cI (repressor) gene were induced by gamma-ray irradiation of lambda phage and of prophage, and 121 mutations were sequenced. Two-thirds of the mutations in irradiated phage assayed in recA host cells (no induction of the SOS response) were G:C to A:T transitions; it is hypothesized that these may arise during DNA replication from adenine mispairing with a cytosine product deaminated by irradiation. For irradiated phage assayed in host cells in which the SOS response had been induced, 85% of the mutations were base substitutions, and in 40 of the 41 base changes, a preexisting base pair had been replaced by an A:T pair; these might come from damaged bases acting as AP (apurinic or apyrimidinic) sites. The remaining mutations were 1 and 2 base deletions. In irradiated prophage, base change mutations involved the substitution of both A:T and of G:C pairs for the preexisting pairs; the substitution of G:C pairs shows that some base substitution mechanism acts on the cell genome but not on the phage. In the irradiated prophage, frameshifts and a significant number of gross rearrangements were also found. PMID- 2966758 TI - [B suppressor cells]. PMID- 2966757 TI - High-level production of hepatitis B viral X protein in Escherichia coli using gene II promoter of bacteriophage M13. AB - Region X is one of the four open reading frames (ORFs) of hepatitis B virus (HBV) and encodes a polypeptide of 154 amino acids (aa). A 584-bp BamHI-BglII fragment of the HBV DNA containing the major part of ORF X which encodes 145 aa was inserted into the BglII site within the gene II of bacteriophage M13. The insertion resulted in an in-phase gene II-X fused protein of 174 aa under the control of the gene II promoter. Cells harboring plasmids (pML alpha X.59 and pMLX.12d) derived from the above construct overproduced the 19-kDa fused protein in Escherichia coli at a level of 10%-20% of total cellular protein. The fused protein was recognized by the anti-X antibodies. This is the first demonstration of using gene II promoter of M13 to express a foreign gene efficiently. PMID- 2966759 TI - Regional enteritis-like enteropathy in a patient with agammaglobulinemia: histologic and immunocytologic studies. AB - A 24-year-old man with agammaglobulinemia developed a form of chronic inflammatory bowel disease over the past 18 years characterized by recurrent diarrhea, malabsorption, and protein-losing enteropathy. In the most recent admission he presented with abdominal cramps and active intestinal bleeding. Radiologic studies showed distal ileal irregularities and strictures that led to two distal intestinal and ileocecal resections. The gross pathologic appearance of these specimens was consistent with regional enteritis. Microscopically, healing ulcers, mucosal irregularities, and a prominent lymphocytic infiltrate without plasma cells or granulomas were observed. Immunocytochemical studies revealed a prominent T-helper cell and a modest T-suppressor/cytotoxic lymphocyte population in the lamina propria. Early and late B-cell differentiation markers were not detected in any of the cells. The immunocytologic findings suggest that T-helper lymphocytes proliferated without inhibition to stimulate non-existent B cells. The study confirms the occurrence of a regional enteropathy-like lesion in the total absence of B-cell function. PMID- 2966760 TI - Linkage between the loci for the Lp(a) lipoprotein (LP) and plasminogen (PLG). AB - A locus, LP, that determines quantitative variation of Lp(a) lipoprotein phenotypes is linked to the plasminogen (PLG) locus (peak lod score = 12.73). This linkage relationship assigns a locus with alleles that have an affect on risk for coronary artery disease to the long arm of chromosome 6. PMID- 2966761 TI - beta-Amyloid gene is not present in three copies in autopsy-validated Alzheimer's disease. AB - Recently, it has been suggested that Alzheimer's disease is associated with a duplication of the amyloid precursor protein gene localized to chromosome 21q21. In this study, a cloned DNA probe (B2.3), complementary to the sequence coding the beta-amyloid peptide, and DNA polymorphisms adjacent to this sequence were used to determine the number of copies of the beta-amyloid gene in DNA isolated from human blood and brain. Individuals with trisomy 21 (Down syndrome) who were heterozygous for the polymorphisms showed a gene-dosage effect, with one allele exhibiting twice the autoradiographic intensity as the other. Heterozygous individuals with Alzheimer's disease and controls showed equal intensities of the two allelic bands, suggesting that there are only two copies of the beta-amyloid gene in these individuals. In individuals with Alzheimer's disease and in controls who were homozygous for these polymorphisms, the number of copies of the beta-amyloid gene was determined by comparing the autoradiographic intensity of beta-amyloid alleles to that of DNA fragments detected by a reference probe. No difference was detected between these two groups. PMID- 2966762 TI - The role of helper T cell products in mouse B cell differentiation and isotype regulation. PMID- 2966763 TI - The role of IL4 and IL5: characterization of a distinct helper T cell subset that makes IL4 and IL5 (Th2) and requires priming before induction of lymphokine secretion. PMID- 2966764 TI - Regulation of cellular and humoral immune responses to collagen type I or collagen type II. AB - We have investigated the characteristics of antigen-specific reductions in murine immune responses to rat collagen type I (R-CI), chick collagen type II (C-CII) or bovine collagen type II (B-CII). Intravenous pretreatment with the appropriate soluble collagen or collagen-coupled spleen cells led to the development of antigen-specific reduced immune responses, the former treatment being more effective than the latter. In the case of CII, pretreatment with R-CI or non related antigens was ineffective. However, pretreatment with denatured bovine CII, native bovine-CII or chick-CII led to immune hyporesponsiveness for either the homologous or heterologous CII molecule. A delayed development of the diminished immune responses was observed for the cell-mediated immune response (CMI), as measured by in vivo delayed-type hypersensitivity (DTH), in that no reduction was evident at Day 7 but a significantly decreased response was observed at Day 14. Collagen-specific IgG and IgM antibody responses were consistently reduced by the pretreatment and remained reduced during the study period. The antigen-specific hyporesponsive state was not sensitive to cyclophosphamide treatment and was not transferable with hyporesponsive spleen cells. Additionally, we have induced unresponsiveness to CII by treating mice with an antibody directed to T helper cells (GK1.5). This treatment led to profound reductions in CII CMI responses as well as CII antibody levels. However, this unresponsive state is not permanent and not transferable with spleen cells from treated mice. These two types of procedures, soluble B-CII i.v. or GK1.5 treatment, not only resulted in CII hyporesponsive states, but also produced delayed onset and decreased incidence of arthritis in the appropriate strains. PMID- 2966766 TI - In vitro generation of adherent mononuclear suppressor cells to Toxoplasma antigen. AB - Adherent mononuclear cells have been found to suppress the lymphocyte proliferation, of T lymphocytes of patients with various chronic infections, to pathogen-specific antigens. To explore mechanisms involved in the generation of these suppressor cells, we established an in vitro method for the generation of suppressor-adherent mononuclear cells. Adherent mononuclear cells separated from mononuclear cells from subjects with serological evidence of chronic Toxoplasma infection could be induced, by preincubation with Toxoplasma antigen for 8 days, to suppress the proliferative response to autologous mononuclear cells to Toxoplasma antigen (TA) (mean suppression = 47%) and tetanus toxoid (TT) (mean suppression = 39%) compared to the proliferative response of autologous mononuclear cells co-cultured with no antigen. When adherent cells were removed after 1-day culture there was no significant suppression of the lympho proliferative response to TA or TT. Induction of the adherent suppressor cell depended on the presence of CD4-positive T cells and not CD8-positive T cells. Adherent suppressor cells acted directly on the proliferative response of CD4 cells to antigen. The adherent cells contained 90 +/- 5% esterase-positive cells. In cell-mixing experiments, equal numbers of CD8-positive T cells pretreated in a similar manner did not have a suppressive effect. However, pretreated CD4 positive cells did have a suppressive effect at higher concentrations of cells than found in the adherent cells. Indomethacin did not alter the suppressive effect. These studies demonstrate the induction of adherent suppressor cells in vitro and implicate the macrophage and CD4-positive T cells as the suppressor cells. PMID- 2966765 TI - Active role of T cells in promoting an in vitro autoantibody response to self erythrocytes in NZB mice. AB - The in vitro anti-self erythrocyte antibody response of NZB spleen cells appears to be influenced directly by T cells. Thy-1+, L3T4+ helper T cells are required for: (i) the generation in vitro of MRBC-specific IgM and IgG AFC by spleen cells from 'autoimmune' (9-12-month old) NZB mice and (ii) the generation in vitro of MRBC-specific IgM and IgG AFC by spleen cells depleted of suppressor cells from pre-autoimmune (2-3-months-old) NZB mice which show no clinical signs of an anti MRBC response. It is evident from the present and previous studies that the anti MRBC autoantibody response is regulated in pre-autoimmune spleen cell populations by Ly2+ T cells. Ly2-T cells from both pre-autoimmune and autoimmune mice in sufficient numbers can overcome this normal regulation and promote the anti-MRBC response in cultures of unfractionated pre-autoimmune spleen cells. Ly2- T cells isolated from autoimmune NZB mice were consistently more active in this than the Ly2- T cells isolated from pre-autoimmune mice, suggesting an enrichment of MRBC reactive Ly2- T cells in autoimmune NZB mice. The Ly2- T cells from autoimmune NZB mice greatly enhance the autoimmune anti-MRBC response relative to a modest enhancement of the response to a foreign antigen, SRBC, produced by the same cells. These data indicate that T cells play an important role both in supporting the autoantibody response to MRBC and in disrupting tolerance, leading to autoimmunity in NZB mice. PMID- 2966767 TI - Peptide hormones and the regulation of sodium excretion. PMID- 2966768 TI - Effect of atrial natriuretic factor on calcium fluxes in adrenal glomerulosa cells. AB - We studied the effect of atrial natriuretic factor (ANF) on calcium influx and efflux in rat adrenal glomerulosa cells stimulated by angiotensin II (Ang II) or potassium ion, and observed how ANF inhibits the initial and sustained phases of the aldosterone response to Ang II or K+ using a superfusion system of dispersed adrenal glomerulosa cells. K+ (8 mM) significantly increased Ca2+ influx rate compared with basal rate (0.91 +/- 0.10 vs 0.42 +/- 0.04 nmol/min/10(6) cells; p less than 0.01). ANF (10(-8) M) did not inhibit the K+-induced increase in Ca2+ influx rate (0.99 +/- 0.18 nmol/min/10(6) cells). Ang II (10(-9) and 10(-8) M) stimulated Ca2+ influx rate (10(-9) M Ang II, 0.62 +/- 0.02; 10(-8) M Ang II, 0.71 +/- 0.09 vs basal, 0.44 +/- 0.03 nmol/min/10(6) cells; p less than 0.05), while ANF (10(-8) M) did not change the Ca2+ influx rate increased by Ang II (ANF + 10(-9) M Ang II, 0.62 +/- 0.06; ANF + 10(-8) M Ang II, 0.69 +/- 0.14 nmol/min/10(6) cells). In the Ca2+ efflux study ANF (10(-8) M) was perfused through the cells 10 minutes before the start of perfusion with Ang II (10(-9) M) or K+ (12 mM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966769 TI - Supernormal contractility in primary hypertension without left ventricular hypertrophy. AB - Forty-three subjects with uncomplicated primary hypertension and without echocardiographic left ventricular hypertrophy and 54 normotensive volunteers were studied by two-dimensional targeted M-mode echocardiography to evaluate systolic function and contractility before the development of compensatory hypertrophy. The ratio of peak systolic pressure to end-systolic dimension was used to assess left ventricular performance and was divided for either posterior wall thickness or cross-sectional area to generate hypertrophy-independent indices of inotropic state. Fractional shortening was normal in the hypertensive group, despite the increase in end-systolic stress. Systolic pressure/dimension ratio was higher in hypertensive subjects (p less than 0.001), as were hypertrophy-independent indices of inotropic state (p less than 0.005), which were inversely correlated to left ventricular mass (p less than 0.001). Values in 11 hypertensive subjects were above the upper confidence limit of the normal shortening/stress relation, which provides a load-independent measure of inotropic state. They showed high hypertrophy-independent indices of inotropic state (p less than 0.01), while the other hypertensive subjects did not. High fractional shortening, wall stress, and systolic pressure (p less than 0.01) were found in the subgroup with supernormal performance, while left ventricular mass was not different from that of other subgroups, depicting inadequate left ventricular hypertrophy. The duration of hypertension was the same in the subgroups. Supernormal inotropic state could be considered one form of primary adaptation to high wall stress that serves to maintain systolic ventricular performance. PMID- 2966770 TI - Atrial natriuretic factor in mild to moderate chronic renal failure. AB - The relationship between kidney function and plasma immunoreactive atrial natriuretic factor (irANF) levels as well as the effects of synthetic human ANF (99-126) were investigated in 13 patients with mild to moderate chronic renal failure. Under basal conditions, glomerular filtration rate averaged 39 +/- 5 (SEM) ml/min/1.73 m2 and blood pressure (BP) averaged 166/107 +/- 7/2 mm Hg; 12 patients were hypertensive. Plasma irANF levels were significantly increased (98 +/- 16 vs 42 +/- 4 pg/ml in healthy control subjects; p less than 0.001) and correlated (p less than 0.05-0.005) inversely with hematocrit (r = -0.65) and positively with systolic BP (r = 0.75) or fractional sodium excretion (r = 0.75). Human ANF-(99-126) infusion for 45 minutes at 0.034 microgram/kg/min augmented (p less than 0.05-0.01) diuresis and urinary sodium, chloride, calcium, phosphate, and magnesium excretion. During the subsequent 45 minutes of human ANF-(99-126) infusion at a rate of 0.077 microgram/kg/min, diuresis and electrolyte excretion remained elevated (p less than 0.05-0.01). Glomerular filtration rate and effective renal plasma flow were not significantly modified, but filtration fraction rose progressively (p less than 0.01). Human ANF-(99-126) infusion decreased BP (p less than 0.05-0.01), produced hemoconcentration (hematocrit + 7%; p less than 0.01) without negative body fluid balance, and increased (p less than 0.01-0.001) plasma norepinephrine, insulin, and serum free fatty acids; plasma aldosterone and renin activity were unaltered during but rose after cessation of human ANF-(99-126) infusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966771 TI - Autologous mixed lymphocyte-tumor reaction and autologous mixed lymphocyte reaction. II. Generation of specific and non-specific killer T cells capable of lysing autologous tumor. AB - The specific and non-specific nature of autotumor cytotoxicity induced in autologous mixed lymphocyte-tumor culture (AMLTC) and autologous mixed lymphocyte culture (AMLC) was studied in patients with carcinomatous pleural effusions. Small- and medium-sized blood lymphocytes that were isolated by centrifugation on discontinuous Percoll gradients did not lyse autologous, freshly isolated effusion tumor cells. In vitro activation of the small lymphocytes, but not of the medium lymphocytes, with autologous tumor cells generated cytotoxic potential restricted to autologous tumor. When stimulated with autologous non-malignant non T cells, the medium lymphocytes, but not small lymphocytes, were triggered to cytotoxicity that acted not only on autologous tumor cells but also on allogeneic tumor cells, T blasts, and tumor cell lines. Experiments using monoclonal antibodies (MAb) and complement (C') showed that both types of killer cells were CD2+ CD3+ CD16- T cells. Autotumor cytotoxicity developed in AMLTC was mediated by the CD4- CD8+ T cell subset in 6 of 9 cases and the CD4+ CD8- subset in the other 3 cases. In contrast, cytotoxicity induced in AMLC was exerted exclusively by the CD8+ subset. The enrichment of blasts from cultured T cells on discontinuous density gradients enhanced autotumor killing activity, with no reactivity recorded for blast-depleted, resting T cells. Addition of mitomycin-C treated large granular lymphocytes (LGL) to AMLTC abolished the induction of autotumor killer cells, whereas non-specific killer cells were generated in AMLC irrespective of the presence of LGL. These results indicate that stimulation of autoreactive T cells in AMLTC and in AMLC could induce 2 distinct types of autotumor killer cells. PMID- 2966772 TI - High tumour prolactin receptor content and lack of increase in serum prolactin levels as predictors of good response to endocrine therapy in rat mammary cancer. AB - Correlations between anti-neoplastic activity of medroxyprogesterone acetate (MPA), on the one hand, and serum prolactin (PRL) levels as well as tumour PRL and insulin receptor content, on the other, were investigated in female rats bearing dimethylbenzanthracene (DMBA)-induced mammary tumours. Changes in liver PRL receptor concentrations were also studied. MPA was injected for 15 days. Regression was observed in 16 out of 50 (32%) tumours from rats treated with MPA. Twenty-seven out of 50 (54%) continued to grow regardless of treatment. Stasis was seen in the remaining 7 tumours (14%). Serum PRL levels increased significantly in rats with tumours which were non-responsive to MPA. Concentration of PRL receptors in the liver of all animals was reduced by MPA treatment. A remarkable increase occurred only in those mammary tumours which responded to therapy. The concentrations of PRL receptors in the tumours non responsive to MPA were similar to those detected in control tumours. Unlike PRL receptors, tumour insulin receptor levels were not modified by MPA treatment. Five out of 14 tumours (35.7%), previously growing in spite of MPA administration, regressed when bromocriptine was added to MPA. A significant reduction in serum PRL levels occurred in all rats undergoing the latter treatment. No difference was observed between responsive and non-responsive animals; on the contrary, the PRL receptor content of responsive tumours increased significantly in comparison with that of non-responsive tumours. PMID- 2966773 TI - Plasma levels of beta-endorphin in uraemic patients during haemodialysis. AB - Plasma levels of beta-endorphins (h-beta-END) are reported to increase under condition of stress. The authors have determined the plasma levels of h-beta-END in six uraemic patients before, during and after haemodialysis. They have found a significant increase at each subsequent time. It is suggested that the cause of the increasing h-beta-END concentrations is the intradialytic stress. PMID- 2966774 TI - Augmentation of the generation of OK-432 activated killer cells after a single dose of mitomycin C in cancer patients. AB - Effect of mitomycin C (MMC) administration on the generation of cytotoxic cells induced by in vitro activation of peripheral blood mononuclear cells (PBM) with OK-432, a bacterial immunopotentiator, was studied in patients with various carcinomas. Following i.v. injection of a single dose of 12 mg/m2 MMC, the ability of PBM to generate OK-432 activated killer cells was markedly increased. Thus, the cytotoxic activity observed 7 days after MMC administration was significantly augmented as compared to that before treatment. Therefore, the ability to generate lymphokine activated killer (LAK) cells was examined, and significantly increased capacity was observed 5 and 7 days after MMC injection. Then, the OK-432 activated killer cell activity significantly correlated with the LAK activity. After treatment, the distribution of lymphocyte subsets exhibited a significant decrease in the percentage of OKT8+ cells. Leu-11+ cells were also reduced. The results appear to indicate that the imbalance in T-cell subsets and the increase in the ability to induce LAK cells may be related to the augmenting effect of MMC administration on the generation of OK-432 activated killer cells in cancer patients. PMID- 2966776 TI - [Dental patients with special physiological and pathological conditions]. PMID- 2966775 TI - Human gamma interferon induction by staphylococcal protein A: effector cells, kinetics and the effect of prostaglandin, indomethacin, ibuprofen and aspirin. AB - Soluble staphylococcal protein A (SpA) induces the synthesis of gamma-interferon (gamma-IFN) by human peripheral blood lymphocytes (PBL). To investigate the kinetics of this gamma-IFN induction and the effector cells involved, we used a highly purified SpA preparation, PBL from healthy volunteers, and a CPE inhibition gamma-IFN assay with Sindbis virus in human fibroblasts. The production of SpA-induced gamma-IFN (SpA-gamma-IFN) peaked 48 h after the addition of SpA to cultures of PBL and decreased after 72 h. Subpopulations of PBL were purified by depletion using specific monoclonal antibodies and complement; CD4+ or OKT4+ (T4: helper/inducer) cells were able to produce SpA gamma-IFN in the absence of CD8+ or OKT8+ (T8: suppressor/cytotoxic) or B-cells. PBL pre-incubated with SpA for more than 72 h inhibited gamma-IFN production by autologous fresh PBL; this inhibition segregated with the T8 subpopulation and was not due to cytotoxicity. SpA-gamma-IFN titers increased markedly when CD3+ or OKT3+ (T3) or T4 cells were incubated with a small number (2-10%) of adherent monocytes, whereas larger numbers (greater than 20%) decreased the yield of SpA gamma-IFN. This decreased yield was probably mediated by prostaglandin E2 (PGE2) of monocyte origin: the presence of PGE2 was demonstrable in these cultures by radioimmunoassay, and the addition of indomethacin reversed the inhibitory effect of large numbers of monocytes; further, treatment of T-cells with exogenous PGE2 also led to an inhibition of SpA-gamma-IFN. Ibuprofen and aspirin also had an effect comparable to indomethacin on SpA-gamma-IFN production. These observations indicate that the production of SpA-gamma-IFN is by T4 lymphocytes, is enhanced by limited numbers of accessory cells (monocytes), and is also regulated by T8 cells via monocyte PGE2. PMID- 2966777 TI - [Clinical evaluation of hemodialysis patients in dentistry]. PMID- 2966778 TI - The fate of immunoreactive opsin following phagocytosis by pigment epithelium in human and monkey retinas. AB - Polyclonal and monoclonal antibodies to human rhodopsin were used to identify and localize this principal glycoprotein of the photoreceptor outer segment discs on thin sections of human and monkey retinal pigment epithelium (RPE) and on immunoblots of RPE subcellular fractions following gel electrophoresis. Antiopsin was visualized with protein A-gold labeling by electron microscopy or peroxidase linked second antibody on immunoblots. In immunocytochemical studies using polyclonal antibodies, the rod outer segments (ROS) were heavily labeled whereas cone outer segments labeling was variable and more sparse. Phagosomes and other small bodies in the RPE, interpreted as secondary lysosomes, were labeled. In contrast, lipofuscin granules, osmiophilic residual bodies of the lysosomal system of the RPE, were negative. No reactive sites were found in Bruch's membrane or in drusen. A monoclonal antibody (MAB) specific for the amino terminus of human opsin and another MAB specific for the carboxy terminal region of bovine opsin produced labeling patterns similar to, but about one-half the density obtained with polyclonal antibodies. In the immunoblot analyses, the lipofuscin granule fraction from a sucrose density gradient of human RPE homogenates was positive for rhodopsin only in those specimens that were found, upon ultrastructural examination, to contain recognizable phagosomes. When phagosomes were lacking and therefore did not contaminate the lipofuscin granule fraction, the immunoblots were negative for opsin. Melanolipofuscin granule fractions were uniformly negative for opsin. We conclude that the superficial hydrophilic antigen binding sites on the opsin molecule for which the antibodies are specific have been altered or destroyed by lysosomal enzyme digestion within the phagolysosomal system of the RPE prior to formation of definitive lipofuscin granules. Thus, these antibodies are of limited value in revealing the ultimate fate of the whole rhodopsin molecule, eg, the hydrophobic sequences that are the most likely residues in lipofuscin granules. PMID- 2966779 TI - AIDS dental treatment program. PMID- 2966780 TI - Comparison of two types of communication methods used after cardiac surgery with patients with endotracheal tubes. AB - In this study we compared two types of communication techniques (planned and unplanned) for effectiveness in communication in the early postoperative intubation period with patients who had cardiac surgery. In this quasi experimental study we used a sample of 40 patients undergoing open heart surgery who participated in preoperative teaching. The control group (n = 20) relied on the experience and creativity of the nurse to provide a method of postoperative communication. The experimental group (n = 20) was introduced to a communication board before surgery and used the board during the postoperative intubation period. Each patient completed an open-ended patient interview, a patient satisfaction questionnaire, and a visual analogue scale to assess satisfaction with communication after discharge from the intensive care unit. An independent t test indicated that a planned method of communication does significantly increase patient satisfaction in the early postoperative intubation period (t = 2.09, p = 0.05, n = 35). Content analyses of the patient interviews further supported this finding. Validity of the patient satisfaction questionnaire was supported by a high correlation (r = 0.70) with results from the visual analogue scale. Nurse satisfaction with communication during intubation was analyzed by use of a nurse satisfaction questionnaire and open-ended questions that were completed by the nurse after caring for the patients in this study. A dependent t test (t = 1.25, not significant, n = 20) and content analysis indicated that planned communication did not increase nurse satisfaction; however, it did add to the repertoire of methods that nurses currently use in trying to communicate with their patients. PMID- 2966781 TI - Preparation and biodistribution of [125I]IBZP: a potential CNS D-1 dopamine receptor imaging agent. AB - Carrier free [125I]IBZP, R-(+)-8-[125I] iodo-2,3,4,5-tetrahydro-3-methyl-5-phenyl 1H-3-benzazepine-7-ol), was prepared from the corresponding uniodinated compound by an oxidative iodination reaction with chloramine-T and sodium [125I]iodide. After purification by column chromatography the desired uptake was obtained with a high purity (greater than 95%). The agent showed good localization in brain after i.v. injection in rats, with an uptake of 2.7, 1.2 and 0.8% dose/organ at 2, 15 and 30 min post injection, respectively. The regional distribution in rat brain, as measured by in vivo autoradiography, displayed a high uptake in the caudate putamen, accumbens nucleus and substantia nigra, regions known to have a high concentration D-1 dopamine receptors. The uptake ratio of striatum/cerebellum increased with time; at 30 s and 2 h after injection the ratio was 1.1 and 5.3, respectively. The specific uptake in the D-1 dopamine receptor regions can be blocked by pretreatment with SCH-23390, a selective D-1 dopamine receptor antagonist (SCH-23390). The corresponding iodine-123 (t1/2 = 13 h, gamma energy 159 keV) labeled agent may be suitable for SPECT imaging of CNS D 1 dopamine receptors. PMID- 2966782 TI - Preparation and biodistribution of [125I]IBZM: a potential CNS D-2 dopamine receptor imaging agent. AB - A new CNS D-2 dopamine receptor imaging agent [125I]IBZM, (S)-3-[125I]-iodo-N-[(1 ethyl-2-pyrrolidinyl)] methyl-2-hydroxy-6-methoxybenzamide, was prepared by either an exchange reaction or by the chloramine-T method. After an i.v. injection, the agent easily passed through the blood-brain barrier and localized in the rat brain. At 2, 15, 30 and 60 min after the injection, the brain uptake was 2.9, 2.3, 1.8 and 0.7% dose/organ, respectively. Regional uptake ratio of striatum/cerebellum (target to nontarget ratio) increased from 1.4 at 30 s to 10.3 at 2 h after the i.v. injection. Digital autoradiography of rat brain sections showed high regional uptake in the caudate putamen and accumbens nucleus, areas known to have a high concentration of the D-2 dopamine receptor. The specific uptake at the D-2 dopamine receptor site was blocked by pretreatment with spiperone, a selective D-2 antagonist. When labeled with 123I (t1/2 = 13 h, 159 keV), [123I]IBZM may be useful for imaging the CNS D-2 dopamine receptor. PMID- 2966783 TI - Comparison of in vivo D-2 dopamine receptor binding of IBZM and NMSP in rat brain. AB - In vivo biodistribution of S- and R-isomers of [125I]IBZM in rats showed a significant initial brain uptake (3.20 and 2.67% dose/organ at 2 min, respectively). The wash-out from the brain was slower for the S-isomer. The striatum to cerebellum ratio for [125I]S-IBZM decreased with an increasing dose of cold carrier or spiperone, suggesting that the brain uptake is stereospecific and saturable, and may be related to the binding of D-2 dopamine receptors. In a dual isotope digital autoradiography study [125I]IBZM and [3H]NMSP(N methylspiperone) show comparable regional cerebral distribution in rats. PMID- 2966784 TI - Effect of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) and dichloromethylidene bisphosphonate (Cl2MBP) on the structure of the organic matrix of heterotopically induced bone tissue. AB - The effect of 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) and dichloromethylidene-bisphosphonate (Cl2MBP) on the structure of the organic matrix of heterotopically induced bone in guinea pig was studied. Heterotopic bone formation was induced by transplantation of allogenic urinary bladder epithelium. Starting from the day of transplantation the animals were treated subcutaneously with HEBP and Cl2MBP with a dose of 12.5 mg P/kg/day during 35 days. The control group was injected with 0.9% NaCl solution. The advantage of heterotopic bone induction as an experimental model is the fact that the applied drugs act on de novo bone formation. Collagen fibers were treated as markers of bone because their size and spatial arrangement reflect the structure and maturity of organic matrix of this tissue. Decalcified histological sections of induced bone, taken 35 days after implantation of inductor, were stained by the picrosirius method. This staining enhances the natural birefringency of collagen fibers and allows for better and specific visualization of collagen fibers bundles under polarizing microscope. In this way the amount of information in the analysed image is increased. Thirty five microphotographs were analysed from each of the investigated groups with the use of optical diffractometry. The radial distribution of light intensity in diffraction patterns was analysed what allowed to evaluate spatial frequencies connected with the width of collagen bundles in induced bone tissue. Since the spatial arrangement of collagen fibers in newly formed bone is random, analysis of angular distribution of light intensity in diffractograms was not performed. Using discriminant analysis the significant differences between all three studied groups of animals were found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966785 TI - Immunohistochemical demonstration of embryonic myosin heavy chains in adult mammalian intrafusal fibers. AB - Serial cross sections of rat, rabbit and cat intrafusal fibers from muscle spindles of normal adult hindlimb muscles were incubated with a monoclonal antibody against embryonic myosin heavy chains. Intrafusal fiber types were identified by noting their staining patterns in adjacent sections incubated for myofibrillar ATPase after acid or alkaline preincubation. In rat and rabbit muscle spindles dynamic nuclear bag1 fibers reacted strongly at the polar and juxtaequatorial regions. Static nuclear bag2 fibers reacted weakly or not at all at the polar region, but showed a moderate amount of activity at the juxtaequator. At the equatorial region both types of nuclear bag fibers displayed a rim of fluorescence surrounding the nuclear bags, while the areas occupied by the nuclear bags themselves were negative. Nuclear chain fibers in rat and rabbit muscle spindles were unreactive with the specific antibody over their entire length. In cat muscle spindles both types of nuclear bag fibers presented profiles which resembled those of the nuclear bag fibers in the other two species, but unlike in rat and rabbit spindles, cat nuclear chain fibers reacted as strongly as dynamic nuclear bag1 fibers. PMID- 2966786 TI - Distribution of sarcoplasmic reticulum Ca-ATPase and of calsequestrin at the polar regions of rat, rabbit and cat intrafusal fibers. AB - Sarcoplasmic reticulum (SR) Ca2+-pumping ATPase (Ca-ATPase) and calsequestrin (CaS) were visualized by indirect immunofluorescence at the polar regions of adult rat, rabbit and cat intrafusal fibers. The immunohistochemical reaction products were regarded as histochemical markers of the SR and as valid indicators of the distribution of the two Ca2+-sequestering proteins. Static nuclear bag2 fibers displayed lower levels of both Ca-ATPase and CaS than the other two intrafusal fiber types. Nuclear chain fibers presented the highest Ca-ATPase levels and, together with dynamic nuclear bag1 fibers, they also exhibited relatively high amounts of CaS. The level of Ca-ATPase was lower in bag 1 fibers than in nuclear chain fibers, but not as low as in bag2 fibers. The comparatively high levels of Ca-ATPase and CaS seen in nuclear chain fibers coincided with their reported faster contractile speeds compared to nuclear bag fibers. PMID- 2966787 TI - Development of lymphocyte subsets in pigtailed macaques. AB - The early development of eight lymphocyte subsets was determined for pigtailed macaque infants from 0 to 800 days of age using two-color flow cytometry and fluorescein- and R-phycoerythrin-conjugated monoclonal antibodies specific for human leukocyte antigens. Four major lymphocyte subsets in monkeys (B, CD4+ T, CD8+ T, and NK cells) could be further divided using two-color analysis. In neonates, the frequency of lymphocyte subpopulations having surface phenotypes found principally on dense, resting cells (IgD+ B cells, Lp220+ CD4+ T cells, and CD18dull CD8+ T cells) was much higher than subpopulations having phenotypes present principally on buoyant, activated cells (IgD- B cells, Lp220- CD4+ T cells, CD18bri CD8+ T cells). There was a complete absence of two CD18bri CD8+ subsets (CD8dull and CD8bri) during the first 300 days of life. The relative proportion of lymphocyte subsets with resting phenotype decreased with increasing age, while the subpopulations associated with activation gradually increased with age. These findings suggest that during early development immunocompetent cells gradually differentiate into activated lymphocytes. PMID- 2966789 TI - Preplacement low-back screening for high-risk areas. PMID- 2966788 TI - Sensitivity of activated human lymphocytes to cyclosporine and its metabolites. AB - Alloreactive T cells generated as clones from mixed lymphocyte cultures, or propagated from heart or liver transplant biopsies, were tested for secondary proliferation measured in the primed lymphocyte test in the presence of Cyclosporine A and metabolites fractionated from human bile. Significant differences were observed in Cyclosporine A sensitivity between various cell cultures ranging as high as 100-fold. The liver is the primary site of Cyclosporine A metabolism, which yields a number of hydroxylated and N dimethylated derivatives that are eventually secreted into the bile. Bile was collected from adult liver transplant patients on Cyclosporine A therapy and following extraction with diethyl ether, separated by high pressure liquid chromatography. Thirteen fractions were tested for their effect on lymphocyte proliferation in concanavalin A activation, mixed lymphocyte cultures and primed lymphocyte test assays. The strongest immunosuppressive effect was found with fraction 8, which contained metabolite M17, which has a single hydroxylation in position 1. Only three other fractions 9, 10, and 13, which contained metabolites M1, M18, and M21, respectively, exhibited immunosuppressive activity, albeit much lower than that of Cyclosporine A. Differences in Cyclosporine A sensitivity among alloreactive T cells followed similar patterns with Cyclosporine A metabolites. Thus, the assessment of the Cyclosporine A effect must consider differences in drug sensitivity of lymphocytes involved in transplant immunity and the generation of metabolites with immunosuppressive activity. PMID- 2966790 TI - Identification of fluorescent glycopeptide derivatives by two consecutive high pressure liquid chromatographic procedures. AB - Reversed phase high pressure liquid chromatography (HPLC) was used to separate individual components of the complex glycopeptide antibiotic teicoplanin in microgram quantities with gradient elution. Each of eight different fractions was then subjected to a specific and highly sensitive HPLC method, which has been developed for the determination of teicoplanin concentrations in biological specimens. This analytical procedure includes precolumn derivatization with fluorescamine and isocratic elution. The fluorescent teicoplanin derivatives were identified by comparing their retention times in both HPLC procedures. Derivatization resulted in increased hydrophobicity and improved chromatographic separation, but the order of elution of the different compounds was not changed. The antimicrobial activity of the individual underivatized fractions correlated with their respective contents of total teicoplanin A2, whereas the pseudo aglycone A3 appeared less active. Similar techniques have the potential to be applied to other complex glycopeptide antibiotics. PMID- 2966791 TI - Mechanical role of expiratory muscles during breathing in upright dogs. AB - To examine the mechanical effects of the abdominal and triangularis sterni expiratory recruitment that occurs when anesthetized dogs are tilted head up, we measured both before and after cervical vagotomy the end-expiratory length of the costal and crural diaphragmatic segments and the end-expiratory lung volume (FRC) in eight spontaneously breathing animals during postural changes from supine (0 degree) to 80 degrees head up. Tilting the animals from 0 degree to 80 degrees head up in both conditions was associated with a gradual decrease in end expiratory costal and crural diaphragmatic length and with a progressive increase in FRC. All these changes, however, were considerably larger (P less than 0.005 or less) postvagotomy when the expiratory muscles were no longer recruited with tilting. Alterations in the elastic properties of the lung could not account for the effects of vagotomy on the postural changes. We conclude therefore that 1) by contracting during expiration, the canine expiratory muscles minimize the shortening of the diaphragm and the increase in FRC that the action of gravity would otherwise introduce, and 2) the end-expiratory diaphragmatic length and FRC in upright dogs are thus actively determined. The present data also indicate that by relaxing at end expiration, the expiratory muscles make a substantial contribution to tidal volume in upright dogs; in the 80 degrees head-up posture, this contribution would amount to approximately 60% of tidal volume. PMID- 2966792 TI - Development of an animal model for investigating disparate myocardial effects of obesity and hypertension. AB - An experimental model for investigating the disparate effects of obesity and hypertension on the heart was developed by ligation of the aorta of male Sprague Dawley rats made obese through ad libitum feeding. Experimental obesity was associated with an increased body fat and cardiac muscle mass, yet a normotensive systemic arterial pressure. Aortic ligation produced an elevated mean arterial pressure and resting heart rate, whereas body weight was similar to that of normotensive lean control rats. Obesity and hypertension together were associated with a significantly increased percent body fat, mean arterial pressure, and left ventricular mass compared with lean controls, whereas pressure and left ventricular weight were greater than those observed in rats with only obesity or hypertension. Cardiac adaptations corrected for body weight indicated that left ventricular weight increased as a function of body weight and body fat, but hypertension produced left ventricular adaptations independent of these variables. These initial studies indicate an additional contribution of hypertension to the left ventricular adaptations of obesity, and this model could therefore be used in future investigations concerning the cardiovascular effects of the simultaneous occurrence of these separate diseases. PMID- 2966793 TI - Coronary vasodilator reserve, capillarity, and mitochondria in trained hypertensive rats. AB - To test the hypothesis that exercise training can reverse the decrements in coronary reserve, capillary density, and mitochondrial volume density evident during established hypertension, we trained spontaneously hypertensive (SHR) and normotensive (WKY) rats on a treadmill over a 3-mo period. At 7 mo of age we used microspheres to evaluate myocardial perfusion in conscious rats. Exercise training did not alter hypertension or left ventricular hypertrophy but did increase maximal O2 consumption in both SHR and WKY. A decrement in left and right ventricular coronary reserve in SHR, compared with WKY, was indicated by 1) a smaller increment in myocardial perfusion during maximal vasodilation with dipyridamole and 2) a higher minimal coronary vascular resistance per unit mass. Exercise training had no significant effect on any index of myocardial perfusion in SHR or WKY. A 12% decrement in capillary numerical density in the endomyocardium of SHR was not reversed by exercise training. We estimated the volume densities of mitochondria, myofibrils, and sarcoplasm using electron microscopy and point-counting stereology on perfusion-fixed hearts. None of the parameters in either SHR or WKY was changed by exercise training. It is concluded that exercise training does not reverse the decrements in coronary reserve and capillary numerical density associated with hypertension in adult rats. Moreover the previously observed enhancement of mitochondrial volume density due to exercise in young hypertensive rats was not observed in adult SHR. PMID- 2966794 TI - Hypoxia does not increase CSF or plasma beta-endorphin activity. AB - The ability of moderate (30-50 Torr arterial PO2) and severe (less than 30 Torr arterial PO2) hypoxia to generate endogenous opioids that modulate ventilation was studied in unanesthetized goats. Ventilation and its components, arterial blood gas tensions and pH, and plasma and cerebrospinal fluid (CSF) beta endorphin activity were measured before and after 4 h of sustained moderate or severe hypoxia. Ventilation, as expected, increased with hypoxia. There were no significant changes in either plasma or CSF beta-endorphin activity after sustained hypoxia. To rule out elaboration of endogenous opioids other than beta endorphin after hypoxia, naloxone or saline was administered to five of the seven goats exposed to 4 h of severe hypoxia, and their ventilatory responses were compared for 30 additional min of hypoxic breathing. No significant differences in ventilation occurred in the two treatment groups during this time period. We conclude that, unlike increases in airway resistance, moderate and severe hypoxia do not cause the elaboration of endogenous opioids that modify respiratory output in unanesthetized adult goats. The apparent ability of hypoxia to cause elaboration of endogenous opioids in the neonate may represent a maturational phenomenon. PMID- 2966796 TI - Morgan Smith, M.D. 1868-1935. PMID- 2966795 TI - Influence of medium composition and culture conditions on glutathione S transferase activity in adult rat hepatocytes during culture. AB - Glutathione S-transferase (GST) activity was measured in adult rat hepatocytes during either pure culture or coculture with another rat liver cell type in various media. Addition of nicotinamide, selenium, or dimethylsulfoxide, deprivation of cyst(e)ine and the use of two complex media were tested. Whatever the conditions used, after a constant decrease during the first 24 h, GST remained active over the whole culture period (1-2 wk). However, various patterns were observed: GST activity either remained relatively stable to approximately 50% of the initial value or showed a moderate or strong increase. The highest values were found in pure hepatocyte cultures maintained in the presence of nicotinamide or dimethylsulfoxide. Similar changes were observed using 1-chloro 2,4-dinitrobenzene or 1,2-dichloro-4-nitrobenzene as substrates for GST. Addition of 10(-4) M indomethacin resulted in 37 to 60% inhibition of enzyme activity. Thus, these results demonstrate that GST remained expressed during culture but its levels markedly varied depending on the medium composition and type and age of culture. PMID- 2966797 TI - Desipramine and cutaneous reactions in pediatric outpatients. AB - The association of adverse cutaneous effects with administration of the tricyclic antidepressant drug desipramine was systematically reviewed in a clinic population of 205 child psychiatry outpatients over a 3-year period. Transient maculopapular rashes developed in 12 children and adolescents treated with desipramine. The rashes had a benign course, disappearing completely in 2 to 7 days, and were not associated with clinical or chemical hepatitis or any other physical symptoms. PMID- 2966798 TI - Thyroid hormone markedly increases the mRNA coding for sarcoplasmic reticulum Ca2+-ATPase in the rat heart. AB - Previous findings have shown that thyroid hormone markedly increases the speed of diastolic relaxation in the heart. This thyroid hormone-dependent change is also accompanied by an increased Ca2+ pumping ability in the sarcoplasmic reticulum. In an effort to determine the underlying cause of improved Ca2+ transport, mRNA levels of the slow Ca2+-ATPase of the sarcoplasmic reticulum were quantified on Northern blots. In hypothyroid rat hearts, the steady state level of Ca2+-ATPase mRNA was only 36% of control levels, whereas hyperthyroid rat heart mRNA levels were 136% of control. Ca2+-ATPase mRNA responded rapidly to T3, as the mRNA level was significantly increased by 2 h and normalized by 5 h after T3 injection into hypothyroid rats. The well established effect of thyroid hormone on improved myocardial contractility and increased speed of diastolic relaxation may in part relate to specific alterations in the level of the mRNA coding for Ca2+-ATPase, resulting in increased pump units. PMID- 2966799 TI - The binding sites of insulin-like growth factor I (IGF I) to type I IGF receptor and to a monoclonal antibody. Mapping by chemical modification of tyrosine residues. AB - The surface topography of IGF I(insulin-like growth factor I) was investigated by chemical modification of amino acid residues in free IGF I and bound to type I IGF receptor or to monoclonal antibody MAB43. Tyrosine residues were modified either by chloramine-T or lactoperoxidase catalyzed iodination. In the free IGF I molecule, all 3 tyrosine residues, A19 (Tyr-60), B25 (Tyr-24), and C2 (Tyr-31), were iodinated. Monoclonal antibody MAB43 protected IGF I against modification at tyrosine residue A19, and in the type I IGF receptor-IGF I complex, all 3 tyrosine residues were shielded against iodine incorporation. These results allow the prediction of the binding domains in the IGF I molecule. The minimal receptor binding site in IGF I would include amino acid residues B25 to C2 and, possibly, the C-terminal part of the A-domain with tyrosine residue A19. PMID- 2966800 TI - Atrial granules contain an amino-terminal processing enzyme of atrial natriuretic factor. AB - At least three enzymes have been identified in atrial tissue homogenates that are capable of processing pro-atrial natriuretic factor to active atrial peptides. The atrial peptides possess potent natriuretic, diuretic, vasorelaxant, and hemodynamic properties, and their existence has implicated the mammalian heart as an endocrine organ. We have purified and characterized a serine proteinase (Mr approximately equal to 70,000) associated with atrial granules that preferentially hydrolyzes the Arg-Ser bond in the synthetic substrates Gly-Pro Arg-Ser-Leu-Arg, benzoyl-Gly-Pro-Arg-Ser-Leu-Arg, and benzoyl-Gly-Pro-Arg-Ser-Leu Arg-Arg-2-naphthylamide, the Arg-2-naphthylamide bond in the substrate benzoyl Gly-Pro-Arg-2-naphthylamide, and the Arg-Ser bond in a 31-residue substrate (Gly96-Tyr126 peptide) corresponding to residues Arg98-Ser99 in pro-atrial natriuretic factor. The Gly96-Tyr126 peptide contains the putative processing site in pro-atrial natriuretic factor and the sequence for the bioactive peptides. Our results indicate that the minimum processing site sequence is -Gly Pro-Arg-Ser-Leu-Arg-Arg- and that the Ser99-Tyr126 natriuretic peptide is the predominant hydrolytic product. After prolonged incubation or at high enzyme concentrations, the Ser103-Tyr126 natriuretic peptide may also be formed. The Ser103-Arg125 natriuretic peptide was only a very minor product. The doublet of basic amino acids is not the primary processing site in pro-atrial natriuretic factor, but their presence may influence cleavage at the single Arg residue "upstream." Our findings are consistent with the idea that the pro-protein and the processing enzymes are packaged into the secretory granule and in response to the proper stimulus, the pro-protein is processed to the active peptides, probably during the process of secretion. The processing pathway of pro-atrial natriuretic factor is discussed. PMID- 2966801 TI - Lectins modulate the internalization of recombinant interferon-alpha A and the induction of 2',5'-oligo(A) synthetase. AB - After binding to specific cell surface receptors, interferon-alpha (IFN-alpha) along with its receptor is internalized by the cells. However, the physiological significance of the internalization of IFN is not known. We have found that the lectin concanavalin A (ConA), which does not inhibit the binding of 125I-rIFN alpha A, inhibits both the internalization of 125I-rIFN-alpha A and the rIFN alpha A-induced increase in the levels of 2',5'-oligo(A) synthetase mRNA and enzymatic activity in the B lymphoblastoid cell line Daudi. The reduced level of IFN-induced 2',5'-oligo(A) synthetase in ConA-treated cells was due neither to direct inhibition of the enzymatic activity nor to generalized inhibition of protein or RNA synthesis. The dose-response curves were similar for the effect of ConA to inhibit 125I-rIFN-alpha A internalization and 2',5'-oligo(A) synthetase induction. The correlation between the ConA-mediated inhibition of both 125I-rIFN alpha A internalization and 2',5'-oligo(A) synthetase induction suggests that internalization of rIFN-alpha A plays a role in the responses to rIFN-alpha A. However, since ConA inhibits protein mobility in the plasma membrane, it is possible that ConA is also preventing aggregation of IFN receptors or interactions between IFN receptors and signal transducing proteins in the plasma membrane that may be necessary for responses to IFN. PMID- 2966802 TI - Plasminogen activation by tissue plasminogen activator in the presence of stimulating CNBr fragment FCB-2 of fibrinogen is a two-phase reaction. Kinetic analysis of the initial phase of slow plasmin formation. AB - Plasminogen activation by tissue-type plasminogen activator (t-PA) is stimulated by fibrin. In a purified system maximal fibrin-enhanced plasmin formation occurs with a delay after an initial phase of slow plasmin formation (lag phase). In the present study purified stimulating CNBr-fragment FCB-2 of fibrinogen was used, and kinetics of plasminogen activation by t-PA were analyzed with respect to the lag phase. At constant FCB-2 concentration the duration of the lag phase decreased with increasing concentrations of t-PA and plasminogen. During this period the rate of plasmin formation/min increased linearly with time with a slope dependent on the initial concentrations of FCB-2, plasminogen, and t-PA. Plasmin pretreatment of FCB-2 resulted in a dose- and time-dependent shortening of the lag phase, and at plasmin concentrations greater than or equal to 1 nM and preincubation times greater than or equal to 3 min maximal plasmin formation occurred without a lag phase. Kinetics during the phase of maximal and constant plasmin formation were not influenced by plasmin pretreatment of FCB-2. We therefore conclude that maximal t-PA-dependent plasmin formation in a system stimulated by purified FCB-2 requires plasmin modification of FCB-2. PMID- 2966803 TI - In vivo [3H]spiperone binding: evidence for accumulation in corpus striatum by agonist-mediated receptor internalization. AB - The processes of receptor internalization and recycling have been well-documented for receptors for hormones, growth factors, lysosomal enzymes, and cellular substrates. Evidence also exists that these processes also occur for beta adrenergic, muscarinic cholinergic, and delta-opiate receptors in frog erythrocytes or cultured nervous tissue. In this study, evidence is presented that agonist-mediated receptor internalization and recycling occurs at the dopamine receptor in rat corpus striatum. First, the in vivo binding of the dopamine antagonist [3H]spiperone was increased by both electrical stimulation and pharmacologically induced increases of dopamine release. Conversely, depletion of dopamine with reserpine decreased in vivo [3H]spiperone binding, but the same reserpine treatment did not alter its in vitro binding. Second, the rate of dissociation of [3H]spiperone from microsomal membranes prepared from rat striatum following in vivo binding was fivefold slower than its dissociation following in vitro equilibrium binding. Mild detergent treatment, employed to disrupt endocytic vesicle membranes, increased the rate of dissociation of in vivo bound [3H]spiperone from microsomal membranes to values not significantly different from its in vitro bound dissociation rate. Third, treatment of rats with chloroquine, a drug that prevents receptor recycling but not internalization, prior to [3H]spiperone injection resulted in a selective increase of in vivo [3H]spiperone binding in the light microsome membranes. The existence of mechanisms that rapidly alter the number of neurotransmitter receptors at synapses provides dynamic regulation of receptors in response to varied acute stimulation states. PMID- 2966804 TI - Prevention of ischemic and postischemic brain edema by a novel calcium antagonist (PN200-110). AB - The effect of PN200-110, a novel calcium antagonist, on the formation of brain edema was examined with rats using a middle cerebral artery (MCA) occlusion model. PN200-110 was effective in preventing the formation of brain edema in 6-h ischemia and in 3-h reperfusion following 3-h ischemia, which were cases in which great accumulations of calcium were autoradiographically observed. Furthermore, PN200-110 diminished the excessive accumulation of calcium in the MCA area involved. These results indicate that an inhibition of the massive influx of calcium into brain cells by PN200-110 may partially ameliorate cell damage, resulting in prevention of brain edema. PMID- 2966805 TI - High-performance liquid chromatographic assay of the metabolites of thymoxamine. PMID- 2966806 TI - Simultaneous micro-determination of nicotinamide and its major metabolites, N1 methyl-2-pyridone-5-carboxamide and N1-methyl-4-pyridone-3-carboxamide, by high performance liquid chromatography. AB - A simultaneous micro-determination of nicotinamide and its major metabolites, N1 methyl-2-pyridone-5-carboxamide (2-py) and N1-methyl-4-pyridone-3-carboxamide (4 py) by high-performance liquid chromatography is described. The method employs a 7-ODS-L (250 mm X 4.6 mm I.D., particle size 7 microns) column eluted with 10 mM potassium dihydrogenphosphate-acetonitrile (96:4, v/v; pH adjusted to 3.0 by the addition of concentrated phosphoric acid) at a flow-rate of 1.0 ml/min. The UV detector was set at 260 nm. The detection limits for nicotinamide, 2-py and 4-py were 10 pmol (1.22 ng), 2 pmol (304 pg) and 2 pmol (304 pg), respectively, at a signal-to-noise ratio 5:1. Isonicotinamide was used as an internal standard. The technique was applied to the analysis of rat and human urines. The total analysis time was ca. 15 min. PMID- 2966807 TI - Peripheral blood lymphocytes from thermal injury patients are defective in their ability to generate lymphokine-activated killer (LAK) cell activity. AB - We have previously shown that natural killer (NK) cell activity against K562 tumor cells is severely depressed in thermal injury patients. In this study we have investigated whether the low NK cell activity present in peripheral blood lymphocytes (PBL) from thermal injury patients could be enhanced by in vitro culture with interleukin 2 (IL2) and whether PBL obtained from these patients could generate lymphokine-activated killer (LAK) cell activity against NK insensitive tumor targets. NK cell activity in PBL obtained from 12 different patients was greatly enhanced against K562 tumor cells after in vitro culture with IL2 for 3 days. In contrast, PBL obtained from these patients and incubated with IL2 had little to no cytotoxic activity when measured against a number of NK insensitive tumor targets. The failure of PBL obtained from thermal injury patients to generate LAK cell activity was observed regardless of the culture time or the amount of IL2 added to the cultures. PBL from thermal injury patients demonstrated reduced proliferative responses to IL2 and, more importantly, contained suppressor cells which could inhibit the generation of LAK cell activity of normal PBL obtained from control individuals. These results clearly show that in some thermal injury patients NK cell activity can be enhanced by IL2 but these patients are defective in their ability to generate LAK cell activity. PMID- 2966808 TI - Immediate effects of intravenous IgG administration on peripheral blood B and T cells and polymorphonuclear cells in patients with myasthenia gravis. AB - Five patients with myasthenia gravis, who received treatment with intravenous 7S gamma-globulin were monitored for changes in immunological status. Serum immunoglobulin G increased from an average of 1.4 to 4.7 g/dl during the 5-day course of therapy. Specific antibody to the acetylcholine receptor present in three of five patients did not change. A transient decrease in total peripheral blood leukocytes was observed in five patients due to decreases in the absolute number of polymorphonuclear cells and lymphocytes in the circulation. Lymphocyte surface marker studies revealed that the percentage of surface immunoglobulin positive cells increased in all patients from an average of 13 to 26% by day 5 of therapy; however, the percentage of HLA-Dr- and Leu 12 (CD19)-positive B cells did not change. Lymphoid cells positive for the Leu 11 (CD16) marker doubled from an average of 11 to 24% during the 5-day course of therapy. Surface Ig-positive cells and Leu 11 (CD16)-positive cells returned to pretreatment values by 7 days posttherapy. Helper/suppressor cell ratios slowly decreased in all patients from an average of 2.9 to 2.2 by 1 week posttherapy and remained low for several weeks. PMID- 2966809 TI - Combined homozygous factor H and heterozygous C2 deficiency in an Italian family. AB - Three of four children in a family have homozygous (less than 1% of normal) deficiency of factor H of the complement system and both parents, who are first cousins, are heterozygous for the same defect. The father and two of the H deficient siblings also have a partial C2 deficiency. One of the children with combined deficiencies is affected by systemic lupus erythematosus with nephritis. No increased susceptibility to infections has been observed in the family. H deficiency is inherited in an autosomal codominant manner and is independently transmitted from C2 deficiency and HLA haplotypes. In the homozygous state it is associated with very low serum concentrations of B and C3, barely demonstrable as activated molecules. C5 is greatly reduced (less than 5%). Also, properdin and C6 9 are decreased. The findings in this family demonstrate that the occurrence of systemic lupus erythematosus in one of the children affected by a combined deficiency of factor H and C2 raises the question whether this pathology is related to the complete factor H or to the heterozygous C2 deficiency. Complete H deficiency is not necessarily accompanied by overt illness. PMID- 2966811 TI - Sterilization and disinfection procedures. A survey of Georgia orthodontists. PMID- 2966810 TI - Abnormal antibody responses in patients with persistent generalized lymphadenopathy. AB - Persistent, generalized lymphadenopathy (PGL) is a recognized component of human immunodeficiency virus (HIV) infection. We conducted longitudinal studies of B and T cell function in seven homosexual men with HIV infection and PGL. All seven had abnormal antibody-mediated immunity as studied by sequential assessment of in vivo antibody responses after immunization with the T-dependent neoantigens bacteriophage phi X 174 and keyhole limpet hemocyanin (KLH), the T-independent tetradecavalent pneumococcal polysaccharide vaccine, and the recall antigens diphtheria and tetanus toxoid. Compared to HIV-negative heterosexual controls, PGL patients responded with lower antibody titers and, following immunization with phage, failed to develop immunologic memory and to switch from IgM- to IgG isotype antibody. In vitro antigen-induced antibody production was markedly diminished; and some patients showed depressed mitogen responses. There was a correlation between the degree of compromised immunity and the clinical condition; those with the most severe symptoms showed the most extensive immune deficiency. Yet despite obvious immunologic impairment five of the seven men have remained clinically stable over a 3-year follow-up period. PMID- 2966812 TI - Fibronectin degradation products containing the cytoadhesive tetrapeptide stimulate human neutrophil degranulation. AB - We investigated whether adhesive glycoproteins, such as fibronectin or fibrinogen, could function to provide a nidus for neutrophil degranulation. Elastase release in recalcified plasma was normal in afibrinogenemic plasma, but 73% less in plasma depleted of fibronectin. Proteolytic digests of fibronectin, but not intact fibronectin (50-1,000 micrograms/ml), induced a concentration dependent release of neutrophil elastase and lactoferrin. MAbs N293, which recognized the mid-molecule of fibronectin, N294, which was directed toward the 11-kD cell adhesive fragment, and N295, generated against the amino terminal of the 11-kD fragment, inhibited the release of elastase by 7, 24, and 60%, respectively. The cytoadhesive tetrapeptide portion of fibronectin, Arg-Gly-Asp Ser (250-1,000 micrograms/ml), released 1.94 +/- 0.10 micrograms/ml of elastase from 10(7) neutrophils, in contrast to the lack of release by the control hexapeptide, Arg-Gly-Tyr-Ser-Leu-Gly. Plasmin appeared to be the enzyme responsible for fibronectin cleavage, since neutrophil elastase release in plasma that had been depleted of plasminogen was decreased and reconstitution of plasminogen-deficient plasma with purified plasminogen corrected the abnormal release. Plasmin cleaved fibronectin to multiple degradation products, each less than 200 kD. This fibronectin digest released 1.05 microgram/ml of elastase from 10(7) neutrophils. We suggest that the activation of plasminogen leads to the formation of fibronectin degradation products capable of functioning as agonists for neutrophils. PMID- 2966813 TI - Clearance and early hydrolysis of atrial natriuretic factor in vivo. Structural analysis of cleavage sites and design of an analogue that inhibits hormone cleavage. AB - This study examines the clearance and early hydrolysis of atrial natriuretic factor (ANF) in vivo. Radiolabeled ANF was cleared from the circulation of the rat with biphasic kinetics; the majority (90%) of ANF cleared with a t1/2 of 15 s, the remaining peptide was cleared with a t1/2 of 5 min. Microsequence analysis of ANF peptides recovered from the circulation of rats revealed five major degradation products of the intact hormone. The first cleavage occurred between amino acids 12 and 13 of the hormone and would inactivate ANF. Over time, additional fragments of the hormone were generated, including fragments of 6, 7, 21, and 24 amino acids in length. Whole body radioautography of rats injected with [123I]-ANF revealed the kidney as a predominant organ involved in clearance of ANF. Subsequent amino acid sequence analyses of radiolabeled ANF exposed to the kidney in vivo indicated that this organ generated four of the five major hydrolysis products observed in circulation, namely, the 6, 7, 16, and 21 amino acid fragments of the hormone. In an attempt to stabilize ANF in vivo, a synthetic analogue of the hormone was prepared that contained the amino acid analogue, aminoisobutyric acid, substituted at position 13. This analogue completely abolished the in vivo cleavage of ANF at this site. These studies demonstrate the usefulness of a protein chemistry approach in characterizing hormone metabolism in vivo and designing analogues with enhanced in vivo stability to cleavage. PMID- 2966816 TI - Mandated interventions and clinical trials: a time for proof. PMID- 2966814 TI - Pulmonary alveolar type II epithelial cells synthesize and secrete proteins of the classical and alternative complement pathways. AB - The serum complement system is a major mediator of inflammation reactions. Two of the complement proteins, the third (C3) and fifth (C5) components, are precursors of potent phlogistic molecules, C3a and C5a. C5a has potent chemotactic activity and plays an active role in pulmonary inflammation. We present evidence suggesting that several complement proteins, including C5, are synthesized locally in the lung in alveolar type II epithelial cells. Lung tissue from normal mice synthesized and secreted C5 protein similar to the C5 protein in mouse serum, whereas lung tissue from C5-deficient mice did not. Lung tissues from both normal and C5-deficient mice synthesized C3. Rat lung tissue synthesized and secreted C5, as well as C2, C4, C3, and factor B. Cultures of type II cells (95% type II cells, 5% macrophages) regularly synthesized all these proteins. In contrast, cultures of macrophages alone synthesized large amounts of C2 and factor B, and in some experiments C3 and C4, but never C5. The C5 synthesized by the rat cells was slightly larger than serum C5 (200 kD compared with 180 kD) and was not processed to the two-chain molecule seen in serum. Rat lung tissue and purified type II cells contained C5 mRNA with the same molecular mass as the C5 mRNA in rat liver and in mouse lung and liver. Human type II cells also synthesized C5, as well as C2, C4, C3, and factor B. Human pulmonary macrophages synthesized only C2, factor B, and, in some experiments, C3. Synthesis of complement proteins in cells that line the alveolar wall may provide a local source of these proteins for inflammatory responses in the lung. Local synthesis of complement proteins could be regulated independently of the synthesis in the liver. PMID- 2966815 TI - Requirements for the construction of antibody heterodimers for the direction of lysis of tumors by human T cells. AB - We constructed a series of MAb heterodimers consisting of the J5 (anti-common acute lymphoblastic leukemia antigen [CALLA]) antibody and antibodies to a variety of structures present on the surface of activated human T cells, including CD3 antigen (T cell receptor-associated glycoproteins), CD2 antigen (T11/E-rosette receptor), CD25 antigen (IL-2 receptor), and the transferrin receptor. We tested the ability of these heterodimers to direct a CD2 + CD3 + CD8 + CD4 - CD25 + transferrin receptor + MHC-restricted human cytolytic T lymphocyte (CTL) clone to lyse a CALLA + human tumor in vitro. Only heterodimers containing an anti-CD3 antibody or activating antibodies to CD2 could direct the clone to lyse these human tumor targets, even when the clone was additionally activated with anti-CD3 or anti-CD2 antibodies. Our findings may have implications in the design of strategies for the use of such reagents in the treatment of human neoplasia. PMID- 2966817 TI - Dicentric chromosome in the bone marrow of a child with megakaryoblastic leukaemia and Down's syndrome. AB - A two year old girl with Down's syndrome (constitutional karyotype: 47 + 21), presenting with pancytopenia, developed acute megakaryoblastic leukaemia (AMKL). Her bone marrow contained an abnormal clone with a novel dicentric chromosome derived from chromosomes 5 and 7 (karyotype 46, XX, -5, -7, +dic (5;7) (p 13; p 11.2), +21. This case provides further evidence for a connection between chromosome 21 and this unusual form of childhood leukaemia, and raises questions about the loss of short arm material from chromosomes 5 and 7 compared with the more usual monosomy or long arm loss. PMID- 2966818 TI - Artificial caries around restorations in roots. AB - An artificial caries technique was used to produce caries-like lesions in the cavity walls adjacent to microfilled resin restorations with and without dentin bonding agent, and glass-ionomer cement restorations, in the roots of extracted teeth. The lesions had histological characteristics similar to those of natural lesions and to those of the lesions produced by other experimental methods. The zonal pattern of the lesions was examined by means of polarized light microscopy and microradiography, and the depth of the lesions was measured for assessment of the microleakage around the restorations. Three cavity cleaning regimes-water spray alone, 50% citric acid, and a proprietary cleaner-were compared in conjunction with the three restorative methods. Cavity cleaning with the citric acid or the proprietary cleaner, and the use of a fluoride-releasing restorative material, acted to reduce the depths of the lesions. Glass-ionomer cement restorations in cavities cleaned with citric acid showed lesions significantly shallower than those treated with other restorations. The glass-ionomer cement was soluble in the acidic environment but promoted the precipitation of minerals in the lesions. The use of a dentin bonding agent did not affect the depth of demineralization around the microfilled resin restorations. PMID- 2966819 TI - Wear and microhardness of a silver-sintered glass-ionomer cement. AB - Knoop Hardness and pin-and-disc-wear measurements were made on a commercial silver-sintered glass-ionomer cement. The objective was to determine whether the incorporation of a bonded-metal-to-glass filler would enhance durability as determined by the above measurements. As with the previous work on conventional (non-metalized) glass-ionomer cements, the specimens were preconditioned at 37 degrees C in air, water, 0.02 mol/L lactic acid (pH 2.67), and heptane. The influence of these media on the microhardness of the silver-sintered material was about the same as that on the conventional materials. Storing in air produced dehydration, which increased the hardness considerably. Heptane storage increased the hardness less, but this increase is attributed to continued curing during storage. After storage in water, the hardness was essentially unchanged; the influence of increased cure is believed to be offset by softening or plasticization from water uptake. Lactic acid produced a decrease in hardness from chemical dissolution as seen from the SEM observations. In most cases, in particular for the air-stored specimens, the wear resistance was enhanced markedly over that of the conventional materials evaluated previously. The exception was the lactic acid-stored specimens for which little, or no, improvement was observed during early periods of wear. The incorporation of silver appeared to provide lubrication, thus reducing wear. However, catastrophic failure from brittle fracture was still a problem, but its occurrence was less frequent. PMID- 2966820 TI - Marginal sealing of curing contraction gaps in Class V composite resin restorations. AB - When one uses composite resins, the curing contraction, the stiffness of the material, and the strength of the vulnerable dentinal bond are important factors in determining the marginal adaptation of the restoration. Calculations based on these intrinsic material properties have indicated that both bulk placement and incremental placement of the restorative material in the cervical cavity inevitably lead to marginal gap formation. Sealing of this gap with an unfilled low-viscosity resin, directly after the composite resin is cured, may lead to perfectly closed restoration margins, provided that composites with low linear curing contraction and low Young's modulus are used. These conditions were experimentally shown to be valid for laboratory and clinical situations. PMID- 2966821 TI - Assessing composite resin wear in primary molars: four-year findings. AB - Many investigators have reported minimal wear of resin restorations in primary molars. Until recently, quantitative wear assessments have not been reported. The purpose of this investigation was (1) to present and contrast data from two 48 month clinical trials wherein quantitative wear assessments were used to evaluate wear of resin restorations in primary molars, and (2) to compare these results with those using the USPHS method of wear assessment. The data were collected from two different clinical trials. One was conducted at the University of California in San Francisco by Tonn and Ryge (TR Trial). The other was conducted at the University of North Carolina at Chapel Hill by Oldenburg, Vann, and Dilley (OVD Trial). The OVD Trial had a sample size of 45 patients with 106 restorations; the TR Trial had 44 patients with 96 restorations. The restorations in both trials were Class I and II restorations in occlusion. They were placed with the experimental light-cured posterior composite resin F-70, later marketed as Ful-Fill. At baseline, six, 12, 24, 36, and 48 months, restorations were evaluated by two evaluators trained in USPHS criteria as well as by a quantitative method of wear assessment, the standard cast technique as described by Leinfelder. Interstudy comparisons were made by Chi-square tests (USPHS evaluations) and ANOVA (quantitative assessments). The results showed that resin restorations in primary molars exhibited progressive wear with time in service. The rate and quantity of wear were very similar to those seen in permanent posterior teeth over 36 months. Findings agree with previous reports that the USPHS evaluation criteria are insensitive in detecting early wear in primary molars. PMID- 2966822 TI - Inhalant allergens as contactants in patients with atopic dermatitis. PMID- 2966823 TI - Assessment of depression in Kuwait by principal component analysis. AB - One hundred depressed inpatients were examined by the WHO schedule for Standardized Assessment of Depressive Disorders (SADD). A common core of symptoms is shared with patients in other studies from Western, Middle-Eastern and international studies. However, pathoplastic cultural influences are manifest in a number of symptoms, notable among which are metaphorical descriptions of symptom intensity by the overwhelmed patient, infrequency of feelings of hopelessness and suicidal attempts, masking of guilt feelings by a front of somatization and a linkage of body weight and sexual functions to health in general. Evidence is provided for a continuum-type unimodal distribution of the principal components studied. PMID- 2966824 TI - The importance of Axis II in patients with major depression. A controlled study. AB - Using a naturalistic study design, we compared 76 depressed patients having a DSM III personality disorder (PD) to a control group of 152 depressed patients with no personality disorder. The patients with major depressive disorder (MDD) and a PD were more likely to have had a younger age of onset, to have had prior hospitalizations, to have a longer duration of episode, to have reported more suicidal thoughts, and to have had more suicide attempts both before and after discharge. Patients with MDD and PD were also more likely to have a family history of alcoholism or antisocial personality and less likely to have dexamethasone nonsuppression, although the latter was not statistically significant. Patients with MDD and PD were less likely to receive electroconvulsive therapy (ECT), equally likely to have received antidepressants, and more likely to receive neither ECT nor antidepressants. Patients with MDD and PD had a poorer response to 'adequate' antidepressants, but a similar response to ECT and 'inadequate' antidepressants. Overall, 91 (60%) of MDD patients and 32 (42%) of MDD plus PD patients were recovered at hospital discharge (X2 = 6.43, P less than 0.025). We conclude that the presence of PD in patients with MDD is associated with a different clinical presentation, family history, and poor recovery at hospital discharge. PMID- 2966825 TI - Clomipramine for panic disorder: I. The first 10 weeks of a long-term comparison with imipramine. AB - Clomipramine and imipramine treatments were compared in a sample of 152 panic disorders. Diagnosis was according to the positive criteria of DSM-III-R, but without exclusion of comorbid affective or personality disorders. The 2-year design provides non-blind treatment under typical clinical practice conditions, and it includes random assignment, periodic assessment with standardized measures, and comparable, flexible drug dosages. Findings on six outcome measures in the first 59 cases to complete 10 weeks showed both tricyclics to be markedly and equally effective for blocking panic attacks, alleviating phobic avoidance, and reducing nonspecific aspects of anxiety. Clomipramine's predominantly serotonergic action seemed not to determine a different action spectrum. During the first 2 weeks, clomipramine was significantly and unexpectedly superior to imipramine in both antipanic and antiphobic actions. These results require replication under double-blind conditions. PMID- 2966826 TI - Lithium prophylaxis of schizoaffective disorders: a prospective study. AB - A prospective study was carried out in order to provide an answer to the following questions. (1) Is lithium effective as a prophylactic agent in broadly defined schizoaffective disorders? (2) Taking for granted that schizoaffective disorders represent a heterogeneous group of conditions, in which schizoaffective patients is lithium effective? (3) Are there any clinical, historical or biological predictors of response to lithium prophylaxis in schizoaffective patients? (4) What are the minimum plasma lithium levels required for effective prophylaxis in schizoaffective disorders? The study confirmed the efficacy of lithium prophylaxis in broadly defined schizoaffective disorders, but showed that this treatment is relatively ineffective in schizoaffective patients with a prominent schizophrenic-like component in their clinical picture and in those diagnosed cross-sectionally as schizodepressive. The only successful predictor of response was a previous bipolar course of the illness (which was associated with a positive outcome of prophylaxis). Plasma lithium levels in the range of 0.45 0.60 mEq/l did not prove to be useful for prophylactic purposes in schizoaffective disorders. PMID- 2966827 TI - The effects of dieting and weight loss upon the stimulation of thyrotropin (TSH) by thyrotropin-releasing hormone (TRH) and suppression of cortisol secretion by dexamethasone in men and women. AB - The effects upon basal hormone levels and neuroendocrine responses of a weight reducing diet allowing 1200 kcal daily were determined in male and female volunteers. Thyrotropin (TSH) responses to thyrotropin-releasing hormone (TRH) were unchanged in men but attenuated in women; this effect was associated with a fall in basal TSH in women, not in men. Rates of non-suppression of cortisol in response to oral dexamethasone (1 mg) were unchanged during dieting although basal morning cortisol levels rose in males and females. The implications for the use of the TRH test and the dexamethasone suppression test (DST) in depressive illness are discussed. PMID- 2966828 TI - Familial subtypes of depression: a longitudinal perspective. AB - An 11-year follow-up of depressed women systematically interviewed and classified by familial subtypes during hospitalization tended to confirm findings of a gradation in severity of illness with depressive spectrum disease being a milder form. Depressive spectrum subjects had shorter subsequent episodes of depression than the other depressives and were less likely to require hospitalization for depression. Depressive spectrum subjects were at greater risk than the others for the development of alcoholism and they experienced more social maladjustment, which suggests implications for treatment strategies and future research. PMID- 2966829 TI - Lymphocyte glucocorticoid receptor binding in depression: normal values following recovery. AB - The number of glucocorticoid receptor sites in lymphocytes and plasma cortisol concentrations were measured in 20 patients who had recovered from major depressive disorder and 20 healthy control subjects. The number of glucocorticoid receptor sites in lymphocytes from the recovered depressed group was not significantly different from that of the control group. Although the mean plasma cortisol concentration in recovered depressives was higher than in control subjects, the difference only just reached significance. This study shows that the reduction in glucocorticoid receptor numbers which occurs during acute depressive illness does not persist on recovery and is, therefore, state dependent. PMID- 2966830 TI - The Diagnostic Melancholia Scale (DMS): dimensions of endogenous and reactive depression with relationship to the Newcastle Scales. AB - The two diagnostic Newcastle Scales for depression have been evaluated in a drug trial with antidepressants. By use of latent structure analysis (Rasch models) it was found that two dimensions are necessary for describing the diagnosis of depression, one for endogenous features and one for reactive features. Of the depressed patients 50% had a pure endogenous depression, 14% had a pure reactive depression, 32% had mixed endogenous and reactive depression, and 4% had uncertain diagnosis. In the pure endogenous depression group 77% had a monotonically non-decreasing improvement curve during treatment whereas in the other diagnostic categories around 50% had such an improvement. PMID- 2966831 TI - MAOI treatment response: multiaxial assessment. AB - While studying the effectiveness of the MAOI isocarboxazid for the treatment of depression, we noted that many patients experienced a reduction of symptoms without equivalent improvement in other areas of their lives. We evaluated four outcome areas: symptoms, work, family functioning and social functioning. After 6 weeks on medication, symptoms improved the most, significantly more so than the other three areas. For the group of patients who completed 24 weeks on medication, all four outcome areas were further improved compared to the 6-week levels, with the improvement in work functioning reaching statistical significance. We conclude that the assessment of treatment outcome is more complex than the simple measurement of symptom reduction, and that different outcome areas are likely to improve at different rates and to different extents. PMID- 2966832 TI - Affective changes with estrogen and androgen replacement therapy in surgically menopausal women. AB - Otherwise healthy surgically menopausal women who had been receiving either an intramuscular estrogen-androgen (E-A) combined preparation or estrogen alone (E) long-term had more positive moods than an untreated control group (CON) coincident with their higher levels of circulating estradiol (E2). Women who received both E2 and testosterone (T) felt more composed, elated, and energetic than those who were given E alone. These findings confirm that mood covaries with circulating estradiol levels in generally healthy, nondepressed women. Possible mechanisms of action of the sex steroids on affect are discussed. PMID- 2966833 TI - The unipolar-bipolar depressive dichotomy and the relationship between afternoon prolactin and cortisol levels. AB - Afternoon prolactin and cortisol levels were measured in 29 patients suffering from a current major depressive episode. Among the 15 unipolar depressed patients the afternoon prolactin and cortisol levels were positively correlated, but 14 bipolar depressed patients did not show a similar relationship, and had prolactin levels lower than the unipolar patients. This finding adds to the growing list of ways in which the neurobiology of bipolar and unipolar depression may differ. PMID- 2966834 TI - Survival and cardiac event rates in the first year after emergency coronary angioplasty for acute myocardial infarction. AB - One year survival and event-free survival rates were analyzed in 342 patients with acute myocardial infarction who were consecutively enrolled in a treatment protocol of early intravenous thrombolytic therapy followed by emergency coronary angioplasty. Ninety-four percent of the patients achieved successful reperfusion, including 4% with failed angioplasty whose perfusion was maintained by means of a reperfusion catheter before emergency bypass surgery. The procedural mortality rate was 1.2% and the total in-hospital mortality rate was 11%. Ninety-two percent of surviving nonsurgical patients who underwent repeat cardiac catheterization were discharged from the hospital with an open infarct-related artery. The related cumulative 1 year survival rate for all patients managed with this treatment strategy was 87%, and the cardiac event-free survival rate was 84%. The 1 year survival for hospital survivors was 98% and the infarct-free survival rate was 94%. Multivariable analysis identified the following factors as independent predictors of subsequent cardiovascular death: cardiogenic shock, greater age, lower ejection fraction, female gender and a closed infarct-related vessel on the initial coronary angiogram. Among patients with cardiogenic shock, despite a 42% in-hospital mortality rate, only 4% died during the first year after hospital discharge. Similarly, the in-hospital and 1 year postdischarge mortality rates were 19 and 4%, respectively, for patients with an initial ejection fraction less than 40, and 25 and 3%, respectively, for patients greater than 65 years. An aggressive treatment strategy including early thrombolytic therapy, emergency cardiac catheterization, coronary angioplasty and, when necessary, bypass surgery resulted in a high rate of infarct vessel patency.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966835 TI - The long-term investment of coronary angioplasty for acute myocardial infarction. PMID- 2966836 TI - Coronary angioplasty: a therapeutic option for symptomatic patients with two and three vessel coronary disease. AB - Coronary angioplasty is a widely applied revascularization procedure for patients with multivessel coronary artery disease. However, follow-up in this patient subgroup is relatively limited. From 1983 to 1986, coronary angioplasty was performed in 349 and 121 patients with, respectively, two- and three-vessel coronary disease with a primary success rate of 83 and 88%. The in-hospital mortality rate was 2.8% (13 of 470 patients). Complete revascularization was achieved in 128 patients. Among the 397 patients with a successful outcome, 373 (94%) were followed up greater than or equal to 1 year; 79% were free of death, nonfatal myocardial infarction or the need for coronary bypass grafting, and 82% of patients had symptomatic improvement by at least one angina functional class. A second coronary angioplasty procedure was required in 13% of patients. After a mean follow-up period of 27 months, an increased incidence of coronary bypass grafting was noted in patients with incomplete versus complete revascularization (16 versus 7%, p less than 0.05). Among the 222 patients who had repeat cardiac catheterization performed an average of 7 months after angioplasty, 103 were symptomatic; 50% of the 222 patients had at least one vessel with greater than or equal to 50% restenosis and 14% of patients had multiple restenoses. In conclusion, coronary angioplasty can be performed with a high initial success rate and marked symptomatic improvement in patients with multivessel coronary disease. However, in this group's experience, the majority of patients selected for coronary angioplasty with multivessel coronary disease will have incomplete revascularization that can be predicted in the majority of patients before the procedure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966837 TI - Percutaneous transluminal coronary angioplasty in two and three vessel coronary disease: information and speculation. PMID- 2966838 TI - Incomplete revascularization in multivessel percutaneous transluminal coronary angioplasty: the role for stress thallium-201 imaging. AB - Evaluation of patients with multivessel coronary disease for percutaneous transluminal coronary angioplasty raises the question: Is incomplete revascularization an acceptable procedure in these patients, or does complete revascularization need to be performed, as in coronary artery bypass grafting? To provide an answer the present study utilized exercise thallium imaging as a guide to the performance of angioplasty in 85 patients with multivessel coronary disease. Preangioplasty exercise thallium imaging helped to identify the primary stenosis ("culprit lesion") in 93% of patients. Two weeks to 1 month after dilation of this lesion, repeat thallium imaging identified two patient groups: Group 1, 47 patients with no evidence of ischemia in a second vascular distribution and Group 2, 38 patients with evidence of further angioplasty. In Group 2 47% of patients had angioplasty of a second vessel and 79% required multivessel angioplasty at 1 year follow-up. In contrast, only six Group 1 patients (13%) required angioplasty of a second vessel at 1 year. Thus, incomplete revascularization may be an acceptable approach in many patients with multivessel coronary disease. Stress thallium-201 imaging may be a useful technique in the evaluation and management of these patients. PMID- 2966839 TI - Responsiveness of atrial natriuretic factor to reduction in right atrial pressure in patients with chronic congestive heart failure. AB - In patients with congestive heart failure, atrial natriuretic factor may serve as a counter-regulatory hormone, offsetting the vasoconstrictive and volume retentive effects of the sympathetic nervous system, the renin-angiotensin aldosterone system and vasopressin. Indeed, the plasma levels of atrial natriuretic factor and the vasoconstrictor hormones are often simultaneously elevated in these patients. It is not known, however, whether atrial natriuretic factor remains responsive to sudden reductions in atrial pressure in patients with chronic heart failure, or is unresponsive like the vasoconstrictor systems. To examine this issue, the plasma concentrations of atrial natriuretic factor and the vasoconstrictor hormones were measured in 20 normal subjects and 12 patients with chronic congestive heart failure during incremental lower body negative pressure, an intervention that lowers atrial pressure. In the normal subjects, incremental lower body negative pressure at -10, -20 and -40 mm Hg decreased central venous pressure and pulse pressure. At maximal lower body negative pressure, plasma atrial natriuretic factor levels decreased from 51 +/- 5 to 27 +/- 3 pg/ml (p less than 0.01), whereas increases occurred in plasma levels of norepinephrine (194 +/- 11 to 385 +/- 70 pg/ml, p less than 0.01), renin activity (1.4 +/- 0.2 to 3.9 +/- 0.1 ng/ml per h, p less than 0.01) and vasopressin (1.3 +/- 0.1 to 6.4 +/- 2.4 pg/ml, p less than 0.05). In the patients with congestive heart failure, lower body negative pressure also reduced central venous pressure. Baseline plasma atrial natriuretic factor levels were markedly elevated, averaging 438 +/- 138 pg/ml, and decreased to 317 +/- 87 pg/ml at maximal lower body negative pressure (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966840 TI - Left atrial spontaneous echo contrast in mitral valve disease: an indicator for an increased thromboembolic risk. AB - The incidence of left atrial spontaneous echo contrast was evaluated in 52 patients with isolated or predominant mitral valve stenosis (Group 1) and 70 other patients who had undergone mitral valve replacement (Group 2). All patients were studied by conventional transthoracic and transesophageal two-dimensional echocardiography. Spontaneous echo contrast could be visualized within the left atrium in 35 Group 1 patients (67.3%) (including 7 patients with sinus rhythm) and 26 Group 2 patients (37.1%) (all with atrial fibrillation). Patients with spontaneous echo contrast had a significantly larger left atrial diameter and a greater incidence of both left atrial thrombi and a history of arterial embolic episodes than did patients without spontaneous echo contrast. Association between spontaneous echo contrast and left atrial thrombi and a history of arterial embolization (considered individually or in combination) showed a high sensitivity and negative predictive value. It is concluded that spontaneous echo contrast is a helpful finding for identification of an increased thromboembolic risk in patients with mitral stenosis and after mitral valve replacement. PMID- 2966841 TI - Physical disability and depression: a longitudinal analysis. PMID- 2966842 TI - Interferons-alpha/beta- and -gamma-resistant Friend cell variants exhibiting receptor sites for interferons but no induction of 2-5A synthetase and 67K protein kinase. AB - A number of Friend leukemia cell variants with a interferon-gamma (IFN-gamma) resistant phenotype have been isolated. They appear resistant to the antiproliferative action of IFN-gamma and to the induction of the antiviral state assessed by Friend leukemia virus release and vesicular stomatitis virus yield. Selection was performed via a prolonged exposure to increasing amounts of highly purified recombinant IFN-gamma of wild-type Friend cells or of variant clones thereof already resistant to IFN-alpha/beta (Affabris et al., 1982, Virology 120, 441-452). Only the clones derived from IFN-alpha/beta-resistant variants showed a phenotype fully resistant to IFN-gamma treatment while keeping their previously acquired resistance to IFN-alpha/beta. These cells are not deficient in high affinity receptors for IFN-gamma so that their resistant phenotype appears to be mediated by events distal to binding of IFN-gamma to its receptors. Furthermore, analysis of IFN-induced dsRNA-dependent 2-5A synthetase and 67K protein kinase enzymatic activities, biochemical markers for cellular responses to IFN, showed that both these activities were not induced in IFN-alpha/beta and IFN-gamma resistant clones when treated with either type of IFN. Accordingly, no increased expression of 2-5A synthetase mRNA(s) could be detected by probing poly(A)+ enriched RNA from cells exposed to IFN-alpha/beta or IFN-gamma treatment with murine or human specific cDNAs. On the other hand, no major changes in restriction patterns of 2-5A synthetase gene(s) were observed in these variant cells by restriction endonuclease digestion and Southern blotting. In addition, analysis of 2-5A synthetase mRNA induction, performed on wild-type cells, showed that the kinetic of induction due to IFN-gamma treatment is slower than that obtained with IFN-alpha/beta. PMID- 2966843 TI - [Study on the immune response of mice receiving bone allografts]. AB - The immune response of bone allografts has not been well understood. The present study was performed to examine the MLC response, CML activity, and alloantibody production of inbred mice receiving fresh and frozen bone allografts. A) Fresh allografts: High MLC response was observed in an early phase of the culture, suggesting the activation and proliferation of helper T cells. Percent cytotoxicity of CML assay was 24% on the 9th day after grafting, indicating the production of cytotoxic T cells. High alloantibody titer was observed on the 30th day after grafting, further indicating the production of the cytotoxic alloantibody. B) Frozen allografts: No significant difference was observed between frozen allografts and fresh isografts (control group) in these immunological responses. These results indicated that the antigenicity of the frozen bone decreased greatly so that matching of the major histocompatibility complex may not be required. PMID- 2966844 TI - An update on hepatitis B viral disease. PMID- 2966846 TI - Virus-triggered immune suppression in mice caused by virus-specific cytotoxic T cells. AB - Normal mice infected with 10(5) infectious doses of lymphocytic choriomeningitis virus (LCMV, WE isolate) generated a reduced or no T cell-independent IgM and/or T cell-dependent IgG response to a subsequent vesicular stomatitis virus Indiana (VSV-IND) injection; this transient immune suppression lasted for weeks to months. Connatally infected LCMV-carrier mice or acutely infected T cell deficient nude mice had normal anti-VSV IgM and IgG or IgM responses respectively. LCMV-infected nude mice transfused with helper cell-depleted LCMV specific immune spleen cells were immunosuppressed. Normal mice infected with LCMV but treated with a rat anti-CD8 mAb (that had been shown previously to eliminate cytotoxic T cells in vivo) and then infected with VSV exhibited a normal anti-VSV IgM and IgG response. Since no IFN-alpha or -beta was detected on, or after, day 6 of LCMV infection, neither LCMV alone, nor IFN induced by it caused the observed immune suppression; the presented evidence suggests that LCMV immune CD8+ T cells were responsible for it. It is conceivable that a similar pathogenesis where virus-specific cytotoxic T cells may destroy virus-infected cells essentially involved in an immune response (APC, T helper cells, etc.) may be involved in other virally triggered immune suppression or in AIDS. PMID- 2966847 TI - Age related changes in the concentration of serum keratan sulphate in children. AB - Serum was obtained from 155 children at the time of admission to hospital for elective surgery. The concentration of serum keratan sulphate was determined by an ELISA which uses an antibody specific for keratan sulphate, a molecule found predominantly in cartilage. Concentrations of keratan sulphate rise progressively during the first four years of life (0-2: mean = 357 micrograms/l; 2-4: mean = 422 micrograms/l) and then remain high until 12 years of age (mean = approx. 500 micrograms/l). At this time, concentrations drop markedly (13-year olds: mean = 377 micrograms/l; 14-year olds: mean = 318 micrograms/l). After age 15, concentrations continue to fall toward the concentrations found in normal adults. Serum concentrations did not show significant differences with respect to disease category, sex or race but were found to vary, sometimes markedly, from child to child at any one age. The results suggest human cartilage undergoes significant changes in metabolic activities during maturation. Measurements of keratan sulphate concentration in serum may prove useful in studying the biochemical and physiological bases of these changes and in monitoring growth or endochondral ossification during maturation. PMID- 2966845 TI - Distinct molecular forms of human T cell receptor gamma/delta detected on viable T cells by a monoclonal antibody. AB - A second type of TCR molecule has been identified on human and murine T lymphocytes, which involves the protein products of the gamma and delta genes. T lymphocytes bearing this receptor may constitute a separate cell lineage with a distinct immune function. We have produced an mAb, which specifically detects human TCR-gamma/delta in native as well as denatured states, this in contrast to previously used anti-gamma chain peptide sera, which only reacted with denatured protein. The receptor occurs in different molecular forms, with or without interchain disulphide bonds, in which a delta chain may or may not be detected by cell surface iodination. The mAb is reactive with all these receptor forms. Therefore, this antibody could be used to determine the expression of TCR gamma/delta on viable human T lymphocytes. In normal individuals, TCR-gamma/delta was found on a subset composing 2-7% of CD3+ lymphocytes in peripheral blood and 0.1-1.0% in thymus. The majority of these cells do not express the CD4 or CD8 antigens, although a significant percentage of CD8+ cells was found. TCR gamma/delta+ cells in peripheral blood are resting lymphocytes, as judged by ultrastructural analysis. T cell clones with different receptor types can display MHC-nonrestricted cytolytic activity, which is shown to be induced by the culture conditions, most likely by growth factors such as IL-2. This strongly suggests that TCR-gamma/delta does not play a role in target cell recognition in MHC nonrestricted cytotoxicity. The anti-TCR-gamma/delta antibody can specifically induce cytotoxic activity in clones expressing the receptor, but in addition inhibit growth factor induced cytotoxicity, which indicates a regulatory role of the TCR-gamma/delta/CD3 complex in MHC-nonrestricted cytotoxicity. PMID- 2966848 TI - The effects of water-soluble vitamins on the expansion of rabbit blastocysts in vitro. AB - The vitamin requirements for culture of rabbit morulae to expanded blastocysts were examined. Early morulae were cultured for 5 days either in a control complete medium containing all the 11 water-soluble vitamins of F10 culture medium (biotin, pantothenate, choline, inositol, niacinamide, pyridoxine, riboflavin, thiamine, folic acid, B12, and lipoic acid) or in media with each vitamin omitted individually. Blastocyst diameters were measured at the end of culture. The omission of inositol, pyridoxine, riboflavin, and niacinamide resulted in large statistically significant decreases in blastocyst expansion. The omission of B12 resulted in a significant increase in blastocyst expansion indicating that the level present in F10 is toxic to rabbit blastocysts. PMID- 2966849 TI - Lambda transducing phage and clones carrying genes of the cysJIHDC gene cluster of Escherichia coli K12. AB - DNA from each of two specialized transducing lambda phage, lambda dcysJIHD and lambda cysJ, has been analysed by heteroduplex mapping. The segment of the Escherichia coli chromosome carried by lambda dcysJIHD was shown to be large, approximately 18 kb in length, and to replace a large length of lambda DNA, approximately 11 kb, which includes the genes for integration and recombination. Thus lambda dcysJIHD is a bio-type transducing phage. lambda cysJ was shown to have lost very little lambda DNA and to carry about 8 kb of bacterial DNA. Sites for several restriction endonucleases were mapped in the DNA from each phage and cloning experiments located some of the genes of the cluster in relation to the restriction map. Cysteine regulation of the cloned cysJ and cysD genes was shown as well as cysteine regulation of beta-galactosidase in some constructs. The direction of transcription of the cysD gene was established, and from physical evidence the size of the 'silent section' between the cysH and cysD genes was estimated to be at least 11 kb. PMID- 2966850 TI - Respiration during sleep in Huntington's chorea. AB - In view of recent reports on lower brainstem dysfunction in Huntington's chorea, we studied respiration during sleep in 12 patients with Huntington's chorea (HC) and in controls. There were no statistically significant differences between patients and controls with respect to apnea periods, respiratory frequency and time elapsed between minimal and maximal value of the respiratory curve. No statistically significant differences in respiratory variability were observed between patients and controls. In the present study, no indication was found for dysfunction of lower brainstem structures involved in respiration in HC. PMID- 2966851 TI - Changes in nocturnal sleep in Huntington's and Parkinson's disease. AB - There have been no reports of studies on nocturnal sleep or quantitative studies of sleep spindle density in Huntington's disease, whereas sleep in Parkinson's disease has been extensively reported. We therefore examined nocturnal sleep changes in patients with Parkinson's and Huntington's disease and compared them with a control group. The previously reported decrease in sleep spindle density was observed in patients with Parkinson's disease. The patients with Huntington's disease, however, had a significant increase in sleep spindle density, the nocturnal sleep pattern being normal in other respects when compared with the control group. This observation may provide a further diagnostic criterion in Huntington's disease. PMID- 2966852 TI - Reflex changes in muscle spindle discharge during a voluntary contraction. AB - 1. This study was undertaken to determine whether low-threshold cutaneous and muscle afferents from mechanoreceptors in the foot reflexly affect fusimotor neurons innervating the plantar and dorsiflexors of the ankle during voluntary contractions. 2. Recordings were made from 29 identified muscle spindle afferents innervating triceps surae and the pretibial flexors. Trains of electrical stimuli (5 stimuli, 300 impulses per second) were delivered to the sural nerve at the ankle (intensity: 2-4 times sensory threshold) and to the posterior tibial nerve at the ankle (intensity: 1.5-3 times motor threshold for the small muscles of the foot). The stimuli were delivered while the subject maintained an isometric voluntary contraction of the receptor-bearing muscle, sufficient to accelerate the discharge of each spindle ending. This ensured that the fusimotor neurons directed to the ending were active and influencing the spindle discharge. The effects of these stimuli on muscle spindle discharge were assessed using raster displays, frequencygrams, poststimulus time histograms (PSTHs) and cumulative sums ("CUSUMs") of the PSTHs. Reflex effects onto alpha-motoneurons were determined from poststimulus changes in the averaged rectified electromyogram (EMG). Reflex effects of these stimuli onto single-motor units were assessed in separate experiments using PSTHs and CUSUMs. 3. Electrical stimulation of the sural or posterior tibial nerves at nonnoxious levels had no significant effect on the discharge of the 14 spindle endings in the pretibial flexor muscles. The electrical stimuli also produced no significant change in discharge of 11 of 15 spindle endings in triceps surae. With the remaining four endings in triceps surae, the overall change in discharge appeared to be an increase for two endings (at latencies of 60 and 68 ms) and a decrease for two endings (at latencies of 110 and 150 ms). The difference in the incidence of the responses of spindle endings in tibialis anterior and in triceps surae was significant (P less than 0.05, chi 2 test). 4. For both triceps surae and pretibial flexor muscles the electrical stimuli to sural or posterior tibial nerves had clear effects on the alpha-motoneuron pool, whether assessed using surface EMG or the discharge of single-motor units. Based on EMG recordings using intramuscular wire electrodes, the reflex effects differed for the gastrocnemii and soleus. 5. In this study, reflex changes in the discharge of human spindle endings were more difficult to demonstrate than comparable changes in the discharge of alpha motoneurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2966853 TI - Light- and electron-microscopical study of phosphoprotein B-50 following denervation and reinnervation of the rat soleus muscle. AB - The neuron-specific phosphoprotein B-50 was originally identified as a phosphoprotein in synaptic plasma membranes isolated from adult brain tissue. In this paper we study the reinnervation of the soleus muscle, a target muscle of sciatic nerve axons, using affinity-purified anti-B-50 antibodies. Light microscopical evaluation of the reinnervation process revealed that the period of muscle fiber reinnervation corresponds closely with the time in which high B-50 immunoreactivity was observed in the nerve fibers that invade the muscle and in the newly formed neuromuscular junctions. Upon completion of reinnervation, B-50 immunoreactivity decreased. In the newly innervating terminals, B-50 was associated with presynaptic vesicular structures and with the presynaptic plasma membrane. In intact mature neuromuscular junctions, virtually no B-50 immunoreactivity could be detected with either light- or electron-microscopic procedures. These observations corroborate the association of high levels of B 50/GAP43 during axon outgrowth and support the concept that B-50 may be a key molecule in the reconstruction of axonal structures. We also observed an unexpected transient increase in B-50 immunoreactivity in the degenerating neuromuscular junctions. This observation cannot be explained in terms of increased neuronal synthesis of B-50, since the degenerating axon processes have been completely disconnected from their cell bodies. Thus, our evidence implies that a rise of B-50 immunoreactivity can be associated with stages of neuronal degeneration as well as with those of neuronal differentiation and axon outgrowth. PMID- 2966854 TI - Immunohistochemical localization of atrial natriuretic peptide in primary aldosteronism. AB - Using the immunoperoxidase method with specific antibody, we examined the presence of atrial natriuretic peptide (ANP) in adrenal tissues of a patient with primary aldosteronism and those derived from a patient with Cushing's syndrome. We found cells immunopositive for ANP in primary aldosteronism but not in Cushing's syndrome. In primary aldosteronism, we noted positively stained cells mainly but not exclusively in the adenomatous tissue. These results demonstrate for the first time the presence of immunoreactive ANP in adrenal tissues of a patient with primary aldosteronism and suggest that adrenal ANP may have potentially important impacts on aldosterone secretion in this disorder. PMID- 2966855 TI - Special education for handicapped and disabled children: classification, programs, and trends. PMID- 2966856 TI - Intrathecal beta-funaltrexamine antagonizes intracerebroventricular beta endorphin- but not morphine-induced analgesia in mice. AB - We have reported previously that beta-endorphin and morphine administered supraspinally produce analgesia by activating different descending pain inhibitory systems in rats. The descending system activated by beta-endorphin involves a spinal endorphinergic system whereas the descending system activated by morphine does not. To determine if this differential action of intraventricular beta-endorphin and morphine also occurs in mice, the effects of pretreatment with intrathecal (i.t.) and i.c.v. beta-funaltrexamine (beta-FNA) on analgesic response induced by i.c.v. and i.t. beta-endorphin and morphine were studied in mice. beta-FNA (2.5 micrograms) was injected i.t. or i.c.v. 24 hr before beta-endorphin or morphine administration and hot-plate and tail-flick responses were measured. Intrathecal beta-FNA attenuated i.c.v. beta-endorphin- but not i.c.v. morphine-induced analgesia. On the other hand, i.t. beta-FNA blocked both i.t. beta-endorphin- and morphine-induced analgesia, but was more effective in blocking the effects of i.t. morphine than beta-endorphin. At the supraspinal sites, beta-FNA administered i.c.v. was found to antagonize i.c.v. morphine-induced analgesia but not i.c.v. beta-endorphin-induced analgesia. The present results in mice are consistent with previous studies in rats and indicate that beta-endorphin and morphine activate different supraspinal opioid receptors. Also, analgesia produced by these two opioids resulted from activation of different descending pain inhibitory systems. The spinal endorphinergic system was involved in the production of i.c.v. beta-endorphin-, but not morphine induced analgesia. PMID- 2966857 TI - Mucosal protective action of histamine against gastric lesions induced by HCl in rats: importance of antigastric motor activity mediated by H2-receptors. AB - The present study was undertaken to investigate the possible mechanism of histamine cytoprotection against 0.6 N HCl-induced gastric lesions in rats by 1) examining functional alterations such as acid secretion, gastric motor activity and mucosal vascular permeability in response to histamine and by 2) comparing the effects of histamine with those of 2-(2-pyridil)-ethylamine (PEA), an H1 agonist and of dimaprit, an H2-agonist. Histamine (3-20 mg/kg s.c.) dose dependently increased acid secretion, inhibited motor activity and reduced the mucosal lesions in response to 0.6 N HCl. Similar effects were observed dose dependently with dimaprit (10-40 mg/kg s.c.), but not with PEA (10 mg/kg s.c.). The protective action of both histamine and dimaprit was attenuated significantly by cimetidine (100 mg/kg s.c.) and indomethacin (5 mg/kg s.c.), but not by tripelennamine (10 mg/kg s.c.), which by itself inhibited significantly motor activity and the lesions. Stimulation of acid secretion caused by histamine as well as dimaprit was antagonized significantly by cimetidine, whereas antigastric motor effects of these agents were decreased significantly by both cimetidine and indomethacin. Histamine and PEA increased significantly the vascular permeability as measured by Evans blue, but the increased vascular permeability caused by 0.6 N HCl was reduced markedly by both histamine and dimaprit. These results suggest that the mucosal protective action of histamine may be mediated at least partly by endogenous prostaglandins through stimulation of H2-receptors, and may be associated with the effect on gastric motor activity but not with that on the mucosal vasculature. PMID- 2966858 TI - Phospholipid N-methylation-dependent alterations of cardiac contractile function by L-methionine. AB - Isolated rat, rabbit and guinea pig hearts exhibited an initial negative inotropic (20-30%) effect followed by a positive inotropic response (60-80%) upon perfusion with 300 microM L-methionine. In contrast, frog hearts did not show any delayed positive inotropic effect, whereas initial negative inotropic effect (25%) of L-methionine was seen. In subsequent studies using rat hearts, methionine was found to induce a dose-dependent increase in contractile force which correlated linearly (r = 0.93) with incorporation of methyl groups into tissue N-methylated phospholipids. The presence of adenosine, L-homocysteine thiolactone and erythro-9-(2-hydroxy-3-nonyl) adenine mixture in the perfusion medium inhibited the contractile effects of L-methionine as well as the incorporation of 3H-methyl groups by about 75%. Cycloleucine, an inhibitor of S adenosylmethionine synthase, and methyl acetimidate, a blocker of the phosphatidylethanolamine polar groups, inhibited phospholipid N-methylation and prevented the contractile changes due to L-methionine. The initial negative inotropic effect of methionine was attenuated by lowering the concentration of Na+, whereas the delayed positive inotropic effect was dependent on the concentration of Ca++ in the perfusion medium. Ryanodine, a blocker of the sarcoplasmic reticular Ca++ release, prevented the positive inotropic effect of methionine whereas verapamil, a well known Ca++ antagonist, blocked the initial depressant effect and reduced the delayed positive inotropic response. Marked alterations in the sarcolemmal and sarcoplasmic reticular calcium transport activities were seen upon perfusing the hearts with methionine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966859 TI - High-resolution polyacrylamide gradient gel electrophoresis (PGGE) of isoenzymes from five Naegleria species. AB - High-resolution polyacrylamide gradient gel electrophoresis (PGGE) was used to separate isoenzymes of 12 Naegleria strains: one N. australiensis, two N. lovaniensis, one N. jadini, two N. gruberi isolated from environmental samples, and six N. fowleri strains isolated from patients with primary amoebic meningoencephalitis. Of the eight enzymes studied, seven showed zymograms with interspecific variation that identified all the species tested. Although the six N. fowleri strains were biochemically the most homogeneous, they showed intraspecific isoenzyme variation that allowed them to be grouped into four zymodemes. The PGGE technique, which separates isoenzymes by their molecular shape, is both sensitive and economical. It offers an addition or an attractive alternative to isoelectric focusing which has commonly been used to aid species identification of Naegleria by separating isoenzymes by their isoelectric point. PMID- 2966860 TI - De novo partial trisomy 15q (proximal type). AB - This report describes a retarded girl with strabismus, high arched palate, antimongoloid slant, low set ears, hearing loss, micrognathia, short neck, and an anteriorly displaced anus. She was found to have a de novo partial trisomy of the proximal part of the long arm of chromosome 15. PMID- 2966861 TI - Unknown syndrome: mental retardation with postaxial polydactyly, congenital absence of hair, severe seborrhoeic dermatitis, and Perthes' disease of the hip. AB - We report a six year old male with mental retardation, postaxial polydactyly and syndactyly, atrichia congenita totalis, severe seborrhoeic dermatitis, recurrent staphylococcal skin sepsis, and Perthes' disease of the hip. His birth may have resulted from an incestuous mating. PMID- 2966862 TI - Transcellular ion currents and extension of Neurospora crassa hyphae. AB - Hyphae of Neurospora crassa, like many other tip-growing organisms, drive endogenous electric currents through themselves such that positive charges flow into the apical region and exit from the trunk. In order to identify the ions that carry the current, the complete growth medium was replaced by media lacking various constituents. Omission of K+ or of phosphate diminished the zone of inward current, effectively shifting the current pattern towards the apex. Omission of glucose markedly reduced both inward and outward currents; addition of sodium azide virtually abolished the flow of electric current. Growing hyphae also generate a longitudinal pH gradient: the medium surrounding the apex is slightly more alkaline than the bulk phase, while medium adjacent to the trunk turns acid. The results suggest that Neurospora hyphae generate a proton current; protons are expelled distally by the H+-ATPase and return into the apical region by a number of pathways, including the symport of protons with phosphate and potassium ions. Calcium influx may also contribute to the electric current that enters the apical region. There seems to be no simple obligatory linkage between the intensity of the transcellular electric current and the rate of hyphal extension. Calcium ions, however, are required in micromolar concentrations for extensions and morphogenesis of hyphal tips. PMID- 2966863 TI - Psychometric analysis of the audiovisual taxonomy for assessing pain behavior in chronic back-pain patients. AB - Sixty chronic back-pain patients were administered the audiovisual taxonomy of pain behavior during their first and last weeks in an inpatient multidisciplinary pain clinic. Audiovisual total score provided a useful index of pain behavior with a suitable frequency and reliability, while offering unique variance as a measure of treatment outcome. Patients' pain behaviors upon admission to the pain program were positively correlated with the following background variables: receiving worker's compensation, pounds overweight, and number of back surgeries. Patients' pain behaviors upon completion of the pain program were significantly correlated with their preferences for pain treatment modalities. High levels of pain behavior correlated with a preference for treatments of ice and heat. Low levels of pain behavior correlated with a preference for physical therapy, social work, lectures, and relaxation. It was suggested that treatment outcome in a multidisciplinary pain clinic is more immediately related to patients' coping styles and their choice of pain treatment modalities than to their demographics and personalities. PMID- 2966864 TI - Sonographic monitoring of ovarian volume during LHRH analogue therapy in women with polycystic ovarian syndrome. AB - Polycystic ovarian disease is characterized by menstrual disorders, infertility, obesity, and large ovaries. Large ovaries with multiple cysts are the direct cause of the high incidence of ovarian hyperstimulation during ovulation induction. Lately, gonadotropin-releasing hormone (GnRH) analogues have been employed to decrease ovarian steroidogenesis and thus reduce the incidence of ovarian hyperstimulation. In this study the ovarian size was ultrasonographically assessed during chronic GnRH analogue treatment, revealing a significant reduction in ovarian volume. This decrease in volume results in a reduced incidence of hyperstimulation, and we think the ultrasonic scanning can be effectively used to assess the success of GnRH treatment. PMID- 2966865 TI - T4+ T helper cell function in vivo: differential requirement for induction of antiviral cytotoxic T-cell and antibody responses. AB - This study documents the differential requirements of T4+ T helper cells in the induction of virus-specific cytotoxic T-lymphocyte (CTL) and antibody responses during acute lymphocytic choriomeningitis virus infection. Two monoclonal antibodies (GK1.5 and RL172.4) directed against the L3T4 (T4) molecule were used for depleting T helper cells from mice. Depletion of T4+ cells caused a pronounced suppression of antiviral antibody response (20-fold decrease) but had minimal effect on virus-specific CTL response (less than 2-fold reduction). Despite the elimination of greater than 90% of T helper cells, anti-L3T4-treated mice were able to generate a CTL response of sufficient magnitude to control the viral infection. In contrast, depletion of Lyt2+ T cells abrogated the CTL response and the ability to eliminate virus. Thus, our results underscore the importance of the Lyt2+ T-cell subset in controlling infection with this virus and show that a deficiency of T4+ T cells is likely to have a more severe effect on antibody production than on CTL responses. PMID- 2966867 TI - Intraoperative duplex scanning after carotid artery reconstruction: a valuable tool. AB - The operative management of extracranial cerebrovascular occlusive disease requires meticulous surgical technique. Intraoperative duplex imaging (10 MHz B mode ultrasound combined with 7.5 MHz Doppler spectrum analysis) was used in an attempt to recognize technical problems that might result in serious neurologic injury. Eighty-four carotid arteries were studied in 76 consecutive patients undergoing carotid endarterectomy or reconstruction. Indications for operation included carotid territory transient ischemic attack (n = 36 [43%]), amaurosis fugax (n = 16 [19%]), stroke (n = 11 [13%]), asymptomatic stenosis (n = 11 [13%]), vertebrobasilar transient ischemic attack (n = 7 [8%]), recurrent stenosis (n = 2 [2%]), and trauma (n = 1 [1%]). Arterial anatomy was assessed ultrasonographically and duplex-derived velocity measurements were obtained to assess hemodynamics. Increased velocity measurement was noted to correlate with ultrasound image of stenosis. Technical errors were identified in 17 patients (22%), including six persons with residual internal carotid artery stenoses, three persons with hemodynamically significant stenoses caused by "kinked" internal carotid arteries, eight persons with debris or intimal flaps in their external carotid arteries, and one person with an occluded external carotid artery. The intraoperative duplex scan led to corrective measures during the surgical procedure in eight patients (11%). No new ipsilateral brain injury was noted in the 76 patients postoperatively. Intraoperative duplex scanning accurately identified residual stenosis, intraluminal thrombus, and loose debris. Duplex scanning altered intraoperative decision making in a significant proportion of patients studied. PMID- 2966866 TI - Use of recombinant retroviruses to study the regulation of integrated adenovirus early promoters. AB - Adenovirus E1A gene products are capable of modulating the expression of a variety of integrated genes. To study the mechanisms by which this regulation occurs, recombinant retroviruses have been utilized to establish cell lines containing an integrated copy of either the adenovirus E2 or E3 promoter adjacent to the bacterial guanine phosphoribosyl transferase (GPT) gene. These cell lines have been characterized with respect to both basal and E1A-induced levels of GPT gene expression. Cell lines with low levels of GPT gene expression showed increased expression in the presence of E1A, whereas cell lines with high basal levels of GPT gene expression had decreased GPT RNA levels in the presence of E1A. Further characterization of these cell lines revealed E1A modulation of the accumulation of RNA initiating at a retrovirus promoter adjacent to the E2 or E3 promoter. The use of the GPT gene as a marker of E2 or E3 promoter activity has allowed the isolation of cell lines which have spontaneously increased their levels of GPT RNA. A preliminary characterization of four of these cell lines has indicated that GPT gene expression is increased as a result of cis activation of the E2 promoter. PMID- 2966868 TI - Mental stress: 'occupational injury' of 80s that even pilots can't rise above. PMID- 2966869 TI - [Screening for antagonistic agents to the lethal toxicity of neocarzinostatin. II. Effects of various drugs in inhibiting the toxicity of neocarzinostatin in vivo]. AB - In clinical chemotherapy with neocarzinostatin (NCS) against cancers, side effects such as leukopenia, anorexia, vomiting and nausea were mainly observed when parenteral administration was used. To prevent these adverse side effects without changing the anticancer activity of the drug, we attempted to apply the two-route-infusion chemotherapy using NCS and antidotes for the NCS treatment devised by Baba. This report presents the results of our study on effects of some antidotes on the acute toxicity of NCS in mice and also on the antitumor activity of NCS against Sarcoma-180 in mice (ICR-JCL strain) when used with tiopronin. The results are summarized as follows. 1. LD50 values of NCS administered via intravenous route increased 2.3- to 3.2-fold when 150, 300, 500 or 1,000 mg/kg of tiopronin was administered subcutaneously together with NCS, 1.3- to 1.4-fold when 50 or 100 mg/kg of sodium thioglycolate was used. When antidotes were given prior to the administration of NCS, 1.8- to 5.4-fold increase in LD50 values of NCS resulted with 300, 500 or 1,000 mg/kg of tiopronin administered 1 hour prior to NCS, 2.3-fold increase resulted with 2,000 mg/kg reduced glutathione, 1.2-fold increase with 100 mg/kg of sodium thioglycolate and 1.9-fold increase with 1,000 mg/kg of L-cysteine monohydrochloride monohydrate. Furthermore, 4.8- to 13.1-fold increase in LD50 of NCS occurred when 150, 300, 500 or 1,000 mg/kg of tiopronin was administered 15 minutes prior to NCS. When these antidotes were administered 1 hour after the administration of NCS, however, no changes in the LD50 value occurred. 2. The LD50 value of NCS given intraperitoneally increased 1.6- to 5.8 fold when 150, 300, 500 or 1,000 mg/kg of tiopronin was administered intravenously at the same time as NCS, 1.4- to 1.6-fold when tiopronin was given 1 hour prior to NCS, intraperitoneally and 1.3- to 1.7-fold when it was given 1 hour after NCS. 3. It was recognized that the acute toxicity of NCS was the most effectively reduced by tiopronin, but only slightly by glutathione, sodium thioglycolate or L-cysteine monohydrochloride monohydrate. The action of tiopronin was the most effective when it was given subcutaneously 15 minutes prior to NCS administered intravenously. 4. The combination chemotherapy on Sarcoma-180 in mice using NCS intraperitoneally and tiopronin intravenously was markedly effective when these agents were given simultaneously. PMID- 2966871 TI - [Evaluation of the effects of beta-lactamase on the concentration of ampicillin in the bronchoalveolar spaces by bronchoalveolar lavage in rats]. PMID- 2966870 TI - Primary hypothyroidism in severe chronic heart failure. AB - Thyroid function was studied in 40 patients with chronic heart failure. Thyroid antibodies and microsome antibodies were negative in all cases. Serum T4, and T3 concentrations showed significant inverse correlation with cardiothoracic ratio, mean right atrial pressure, pulmonary artery systolic pressure, and peripheral venous pressure. Serum T4, T3 concentrations showed significant correlation with PaO2, serum albumin, and serum cholinesterase. Serum TSH concentrations increased with increasing cardiothoracic ratio. Histological examinations showed fibrosis and atrophy of the thyroid gland in 2 cases. These findings suggest the possible development of primary hypothyroidism as a result of chronic heart failure. PMID- 2966872 TI - Effects of atrial natriuretic factor in rats with experimental high-output heart failure. AB - The effects of atrial natriuretic factor (ANF) were evaluated in rats with chronic aorto-caval (A-V) fistula. In this experimental model of high-output heart failure, the animals display elevated atrial pressure and systemic vasodilation, but avidly retain sodium. Experiments were performed on Munich Wistar rats, 8 to 14 days after placement of an infrarenal surgical anastomosis (side-to-side, 0.9 +/- 0.2 mm) and on sham operated controls. Infusion of synthetic ANF (3-28) intravenously (5 micrograms/kg prime; 0.17 microgram/kg.min) resulted in increases in urine flow (V) and fractional sodium excretion (FENa) and decreases in blood pressure (BP) that were significantly attenuated in rats with A-V fistula compared to controls. To control for the lower baseline BP that was present in rats with A-V fistula, a second series of studies was performed in which renal perfusion pressure was reduced in normal rats to 110 mm Hg with a servocontrolled pneumatic cuff. ANF infusion to this group led to significant increases in glomerular filtration rate (GFR), V and FENa that were greater than those seen in rats with A-V fistula (FENa = 2.7 +/- 0.3% vs. 0.48 +/- 0.12%, P less than 0.05). Thus, the moderately reduced BP in rats with A-V fistula did not account for the blunted response to ANF. To investigate whether the renal sensitivity to ANF is altered in this model, an additional series of experiments were performed in which ANF was infused over a range of doses (0.08 to 2.5 micrograms/kg.min) to both groups of rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2966873 TI - Cyclosporine A induced remission of relapsing nephrotic syndrome in children. AB - We treated 20, steroid resistant or steroid dependent and cyclophosphamide or chlorambucil treated, relapsing nephrotic patients with oral cyclosporine A for eight weeks. Cyclosporine A was started at 7 mg/kg/day and titrated to maintain HPLC level of 100 to 200 ng/ml. Of 20 patients, 14 had a complete remission and the remaining 6 had a reduction in their proteinuria. The mean serum albumin of the 14 responders rose from 2.1 g/dl to 4.1 g/dl (P less than 0.00001) after cyclosporine A therapy. The mean serum cholesterol of the 14 responders decreased from 394 mg/dl to 184 mg/dl (P less than 0.0001) after cyclosporine A therapy. The mean creatinine clearance of the 20 patients (104 ml/min/1.73 m2) was unchanged (107 ml/min/1.73 m2) after eight weeks of cyclosporine A. By life table analysis, 40% of the responders show a sustained remission of up to a year. Cyclosporine A responders had a higher T3 cell count prior to therapy compared to nonresponders (69 +/- 5.54% vs. 61 +/- 6.4%, P less than 0.02). Pre-therapy interleukin-2 levels measured in 10 patients were normal or supranormal in 8, 6 of whom were treatment responders. Two patients with low interleukin-2 levels were nonresponders. Cyclosporine A can be used to induce a remission in relapsing nephrotic patients, and short-term cyclosporine A therapy does not produce nephrotoxicity. PMID- 2966874 TI - [Eye manifestations in multihandicapped children with cerebral disorders]. PMID- 2966875 TI - [Laparoscopic diagnosis and laser therapy of tuberculous peritonitis]. PMID- 2966877 TI - An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Pasteurella pneumotropica in murine colonies. AB - An ELISA for the detection of class specific IgG antibodies to Pasteurella pneumotropica was developed for the serological diagnosis of infections in mouse colonies. Heat inactivated whole cell preparations of an isolate of P. pneumotropica biotype Heyl (strain P 166) served as antigen for the ELISA procedure and for immune serum production in germ-free Han:NMRI mice. Cross reactions with the autochthonous flora of Han:NMRI SPF-mice were not observed, but were evident when a P. pneumotropica antiserum was tested against other antigens of the Pasteurella-Actinobacillus group. According to the reclassification of this bacterial group proposed by Mutters et al. (1), strains of the following species were tested: P. anatis, P. canis, P. dagmatis, P. langaa, Pl multocida sub. multocida, P. pneumotropica biotype Jawetz, P. stomatis, Actinobacillus equuli and A. lignieresii. Clear cross reactions could be shown with P. pneumotropica biotype Jawetz and A. equuli and to a lesser extent with P. anatis. Antibody formation profiles after nasal infection of Han:NMRI mice exhibited a primary rise of IgG-type antibody titer between 17 to 21 days post infection. Investigations of different mouse colonies free and infected with P. pneumotropica revealed good correlations between serological and bacteriological findings. PMID- 2966876 TI - [Compression of both lungs by severe right heart dilatation in congenital pulmonary atresia]. AB - Neonatal cyanosis due to pulmonary atresia is seen in one-third of all cyanotic neonates with congenital malformation of the heart. In both types of this disorder survival is dependent on patency of the ductus arteriosus. Consequently the first days of life most often show dramatic changes in clinical status, sometimes similar to that in patients with transposition of the great arteries. Intensive care treatment of the baby, rapid diagnostic management and early operative intervention may improve the poor prognosis. Because of the numerous cardiopulmonary and diagnostic complications, only 50 percent of patients will reach 4 weeks of age if not operated. We describe an unusual case of type 2 pulmonary atresia with intact ventricular septum in which pulmonary complications led to irreversible cardiac failure. Aneurysmatic enlargement of the right atrium caused compression atelectasis of nearly all parts of the lungs, followed by a unilateral pneumothorax after postnatal emergency ventilation. Clinical and circulatory status deteriorated. Successful pharmacological or operative therapy was impossible at this point. We want to stress upon the fact that - despite the hopeless situation of our patient - we were able to establish diagnosis by improvised procedures, which can be performed in every ICU. PMID- 2966878 TI - Naturally occurring subclinical Corynebacterium kutscheri infection in laboratory rats: strain and age related antibody response. AB - Naturally occurring subclinical Corynebacterium kutscheri infection was analyzed by antibody response related to the strain of rats. Wistar-Lewis, Wistar and Spraque-Dawley rats were high responders in seroconversion rates and antibody titers, while Brown Norway and Fischer rats were low responders. The antibody response was related to age also. Some young rats had maternal antibody to C. kutscheri, but antibody disappeared before 8 weeks of age. Rats were antibody negative for several months thereafter and became antibody-positive after 6 months of age. The antibody response was highest at 8 to 9 months of age in subclinical C. kutscheri infection. This antibody response was very late, compared to the antibody response to Sendai virus and Mycoplasma infections. PMID- 2966879 TI - Chronic intrathecal cannulation affects hypothalamic beta-endorphin and met enkephalin concentrations. AB - Beta-endorphin concentrations decrease and met-enkephalin concentrations increase in the hypothalamus of rats bearing a chronic intrathecal cannula. The modification of the concentrations of beta-endorphin is already present on day 1 after surgery, whereas met-enkephalin is affected starting on day 6, and both peptides are still modified on day 10. These results indicate that chronically cannulated rats are not to be considered normal animals. PMID- 2966880 TI - The nature of lysosomal enzyme glycosylation in ALL: relevance to abnormal beta hexosaminidase expression. AB - The glycosylation of beta-hexosaminidase was investigated in the transformed cell line, CCRF/CEM, derived from a human acute lymphoblastic leukaemia, and comparisons were made with enzyme from normal human skin fibroblasts. A series of studies including neuraminidase sensitivity, lectin chromatography, Biogel P4 chromatography of [3H]-mannose-labelled glycopeptides and endoglycosidase susceptibility, provided clear evidence that in CCRF/CEM cells, beta hexosaminidase was abnormally glycosylated. The results indicate that leukemia associated changes in beta-hexosaminidase expression are probably due to increased sialylation of highly-branched complex oligosaccharides. PMID- 2966881 TI - Single-fiber laser Doppler flowmetry. A method for deep tissue perfusion measurements. PMID- 2966882 TI - Quantitation of mu mRNA by in situ hybridization reveals a correlation between B maturation associated antigens and IgM gene activation in acute lymphatic leukemias. AB - Acute lymphatic leukemias, expressing the common acute leukemia antigen were investigated for expression of early T- and B-cell associated markers and the activation of the gene for the heavy chain of the immunoglobulin M. The gene activation as determined by quantification of the mu mRNA with a fluorochrome labeled gene probe in situ in individual cells showed a wide spectrum of positivity which was correlated to increasing expression of B-maturation markers. There was also a correlation between the amount of cellular immunoglobulin as determined by immunofluorimetry. Quantitation of mRNA in individual cells may allow to determine the degree of gene activation in antigenically defined individual cells and may thus contribute a new tool for classification of normal and malignant cells. PMID- 2966884 TI - Nocturnal fluctuations of plasma atrial natriuretic peptide in normotensive and hypertensive men. AB - Plasma levels of immunoreactive atrial natriuretic peptides (ANP) were measured at 10-min intervals during night-sleep in 4 normotensive and in 4 moderate, essential hypertensive subjects. The mean ANP levels ranged from 24.3 to 27.9 pg.ml-1 for the normal subjects. These mean levels were not significantly different in the hypertensive subjects (range: 26.3 to 37.2 pg.ml-1). Fluctuations, often of small amplitude, were observed around this mean, without any defined periodicity. Changes in plasma ANP were not associated with changes in heart rate. Analysis of the ANP profiles and the concomitant sleep stage patterns did not reveal any temporal relationship between ANP fluctuations and specific sleep stages or waking periods. The ANP profiles did not differ between the groups, which indicates no abnormality in ANP secretion in moderate essential hypertension. PMID- 2966883 TI - Altered placental insulin and insulin-like growth factor-I receptors in diabetes. AB - We have compared the characteristics of IGF-I and insulin receptors in placentas of normals and insulin dependent diabetic patients. Specific binding of both IGF I and insulin in placental membranes from patients with good glycemic control (as reflected by blood hemoglobin content) was unaltered while that in the placental membranes from the patients with poor glycemic control was increased to approximately 20% of the normals. This observed small but significant (p less than 0.05) increase in binding of IGF-I and insulin to placental membranes from diabetic patients with poor glycemic control was further magnified, approximately twice (p less than 0.001) the normal, when the membrane receptors were purified by lectin chromatography. The kinetic analysis of IGF-I and insulin binding in both membranes and lectin purified receptors revealed that the increased binding of insulin and IGF-I to the placentas from diabetic patients with poor glycemic control was due to an approximately 2 fold increase (p less than 0.001-0.05) in the receptor numbers without any significant changes of the affinities. The molecular characteristics of the receptors in these diabetic patients, as revealed by the cross-linking studies, did not reveal any changes when compared to the normals. The parallel changes of IGF-I and insulin receptors, shown here, are in accordance with the homologous nature of these two receptors. The increased receptor numbers of these two interrelated hormones in placentas of diabetics with poor glycemic control may be relevant to the altered placental functions in diabetic pregnancy. PMID- 2966885 TI - Enhanced stereotypic behavior by chronic treatment with bromocriptine accompanies increase of D-1 receptor binding. AB - Different effects of chronic treatment with bromocriptine (BRO) on D-1 and D-2 receptors in the rat were studied through behavioral observation and DA receptor binding assays. Chronic BRO led to enhancement of stereotypy to apomorphine (APO), at the same time it increased the density of D-1 receptor binding by 43% and decreased that of D-2 receptor binding by 21%. Our data suggest that BRO has different effects on D-1 and D-2 receptors and the behavioral hypersensitivity caused by BRO may be relevant to the proliferation of D-1 receptor. PMID- 2966887 TI - Anti-idiotypes against a monoclonal anti-haloperidol antibody bind to dopamine receptor. AB - Anti-idiotypic antibodies were raised in rabbits by immunization with a monoclonal anti-haloperidol antibody. Some of these anti-idiotypic antibodies bind in a concentration dependent manner to bovine striatal membranes. Following affinity purification, these antibodies inhibit haloperidol binding to striatal membranes and deplete [3H]-spiperone binding sites from a solubilized preparation of striatal membranes. It is thus concluded that these anti-idiotypic antibodies are an internal image of haloperidol and as such can interact with D2-dopamine receptors. PMID- 2966886 TI - Evidence for dopamine D-2 receptors on cholinergic interneurons in the rat caudate-putamen. AB - The aziridinium ion of ethylcholine (AF64A) is a neurotoxin that has demonstrated selectivity for cholinergic neurons. Unilateral stereotaxic injection of AF64A into the caudate-putamen of rats, resulted in a decrease in dopamine D-2 receptors as evidenced by a decrease in [3H]-sulpiride binding. Dopamine D-1 receptors, labeled with [3H]-SCH 23390, were unchanged. The efficacy of the lesion was demonstrated by the reduction of Na+-dependent high affinity choline uptake sites labeled with [3H]-hemicholinium-3. These data indicate that a population of D-2 receptors are postsynaptic on cholinergic interneurons within the striatum of rat brain. PMID- 2966888 TI - [Scientific organization of work in the operations of a urology department]. PMID- 2966889 TI - Ribosomal DNA inheritance and recombination in Neurospora crassa. AB - The genetic segregation of ribosomal DNA (rDNA) in Neurospora crassa was analyzed by exploiting restriction fragment length polymorphisms in the nontranscribed spacer (NTS) sequences of nine laboratory wild-type strains and wild-collected strains. In an analysis of random spore progeny from seven crosses, and of ordered tetrads from two of those crosses the rDNA was shown to be inherited in a simple, stable Mendelian fashion, exhibiting an approximately 1:1 ratio of the two parental rDNA types. No meiotic recombinants were detected among the progeny, indicating that non-sister-chromatid crossing over is highly suppressed in the rDNA region. The basis for this suppression of meiotic recombination is not known. PMID- 2966890 TI - Skin blood flow from gas transport: helium xenon and laser Doppler compared. AB - A study was designed to compare three independent measures of cutaneous blood flow in normal healthy volunteers: xenon-133 washout, helium flux, and laser velocimetry. All measurements were confined to the volar aspect of the forearm. In a large group of subjects we found that helium flux through intact skin changes nonlinearly with the controlled local skin temperature whereas helium flux through stripped skin, which is directly proportional to skin blood flow, changes linearly with cutaneous temperature over the range 33 degrees to 42 degrees. In a second group of six volunteers we compared helium flux through stripped skin to xenon-133 washout (intact skin) at a skin temperature of 33 degrees, and we found an essentially linear relationship between helium flux and xenon measured blood flow. In a third group of subjects we compared helium flux blood flow (stripped skin) to laser doppler velocimetric (LDV) measurements (intact skin) at adjacent skin sites and found a nonlinear increase in the LDV skin blood flow compared to that determined by helium over the same temperature range. A possible explanation for the nonlinear increases of helium flux through intact skin and of LDV output with increasing local skin temperature is that they reflect more than a change in blood flow. They may also reflect physical changes in the stratum corneum, which alters its diffusional resistance to gas flux and its optical characteristics. PMID- 2966891 TI - Malignant mesothelioma. PMID- 2966892 TI - [Hemotrophogenic fecundity of mass species of black flies (Diptera, Simuliidae) in the center of the European portion of the USSR]. PMID- 2966893 TI - [Identification and initial results of the study of Lyme disease in northwestern USSR]. PMID- 2966894 TI - [Formation of enterobiasis foci at a new building site (Atommash) and their eradication]. PMID- 2966896 TI - The structural gene for a phosphorus-repressible phosphate permease in Neurospora crassa can complement a mutation in positive regulatory gene nuc-1. AB - van+, a gene encoding a phosphorus-repressible phosphate permease, was isolated by its ability to complement nuc-1, a positive regulatory locus that normally regulates van+ expression. This was unexpected because the nuc-1 host already contained a resident van+ gene. Plasmids carrying van+ complemented a nuc-2 mutation as well. Probing of RNA from untransformed wild-type (nuc-1+) and constitutive (nuc-1c) strains by van+ probes indicated that levels of the van+ transcript were subject to control by nuc-1+. Probing of the same RNAs with a cosmid clone, containing approximately 15 kilobases of upstream and downstream DNA, revealed no other detectable phosphorus-regulated transcripts within this 40 kilobase region of the chromosome. PMID- 2966895 TI - A retrovirus expressing the 12S adenoviral E1A gene product can immortalize epithelial cells from a broad range of rat tissues. AB - An epithelial cell-transforming virus could be of great use, both in the culture of epithelial cell lines and in the study of carcinogenesis. Since the adenoviral E1A gene has been shown to partially transform some epithelial cells from primary rat cell cultures, we constructed retrovirus vectors containing either the 12S or 13S E1A cDNA sequences to facilitate the transfer of these genes into a variety of primary cell types. The 12S E1A virus induced proliferation and immortalization of epithelial cells in rat kidney, liver, heart, pancreas, and thyroid primary cultures. In the two cases tested, heart and liver cultures, E1A immortalized cells were nontumorigenic, but could be completely transformed by subsequent introduction of the ras oncogene. To our surprise, the 13S virus had a greatly reduced immortalization potential. We discuss these data in light of the model of Spindler et al. (K. R. Spindler, C. Y. Eng, and A.-J. Berk, J. Virol. 53:742-750, 1985), in which the 12S E1A protein is required for the complete induction of the cellular DNA replication machinery in the quiescent human epithelial cells in which adenoviruses normally replicate. PMID- 2966897 TI - Sonographic identification of Down's syndrome. PMID- 2966898 TI - Effects of acute intermittent exposure to cigarette smoke on hypothalamic and preoptic catecholamine nerve terminal systems and on neuroendocrine function in the diestrous rat. AB - Diestrous female rats were exposed to the smoke from one to four cigarettes. Exposure to unfiltered cigarette smoke produced dose- and time-dependent reductions of catecholamine levels and dose- and time-dependent increase in catecholamine utilization in the various hypothalamic and preoptic dopamine and noradrenaline nerve terminal systems. These effects were counteracted by pretreatment with the ganglion blocking agent mecamylamine (1 mg/kg). Exposure to cigarette smoke was also found to produce a dose- and time-dependent inhibition of serum prolactin, LH and FSH levels which was counteracted by pretreatment with mecamylamine. Exposure to the smoke from one cigarette (but not from four cigarettes) increased serum TSH levels. In combination with tyrosine hydroxylase inhibition the exposure to cigarette smoke produced a dose- and time-dependent inhibition of plasma ACTH levels, an action which was counteracted by pretreatment with mecamylamine. The results demonstrated a sex difference (cf. Anderson et al. 1985c), in the nicotine-induced changes of TSH and ACTH secretion despite a general increase in hypothalamic and preoptic dopamine and noradrenaline utilization in both the male and the diestrous female rat. The differences in the neuroendocrine actions of acute intermittent exposure to cigarette smoke in the diestrous rat and the normal male rat are discussed. PMID- 2966899 TI - Nicorandil shortens action potential duration and antagonises the reduction of Vmax by lidocaine but not by disopyramide in guinea-pig papillary muscles. AB - Conventional microelectrode techniques were used to examine whether or not nicorandil, which shortens action potential duration (APD), modifies the lidocaine- or disopyramide-induced time-dependent reduction of Vmax in guinea-pig papillary muscles. First, effects of 0.1 and 1 mmol/l nicorandil were examined on the frequency dependence of Vmax and on the recovery process of Vmax. Second, the frequency-dependent reduction of Vmax by 20 mumol/l antiarrhythmic drugs was examined in the presence and absence of 1 mmol/l nicorandil at stimulation frequencies of 1/120 Hz - 5 Hz. Third, the recovery process of Vmax in the presence of 20 mumol/l antiarrhythmic drugs was examined, with and without 1 mmol/l nicorandil, by applying test stimuli at various diastolic intervals after conditioning stimuli. 1 mmol/l nicorandil greatly shortened APD90 to 30-40% of control without changing the frequency dependence of Vmax, the recovery process of Vmax, and the resting potential. The lidocaine-induced, frequency-dependent reduction of Vmax was significantly antagonised by 1 mmol/l nicorandil, but the disopyramide-induced reduction was not. The recovery process of Vmax slowed in the presence of lidocaine was antagonised by 1 mmol/l nicorandil as follows: the time to get the full recovery of Vmax was shortened by nicorandil with a significant decrease in the zero time-intercept (from 0.54 to 0.38) but with an insignificant change in the recovery time constant (from 130 ms to 121 ms). In contrast, the recovery process of Vmax slowed in the presence of disopyramide (a zero time intercept of 0.13 and a recovery time constant of 50 s) was not significantly antagonised by 1 mmol/l nicorandil. In conclusion, nicorandil having an action potential-shortening action antagonises the lidocaine-induced, time-dependent reductions of Vmax, but not the disopyramide-induced reductions. These results suggest that: (1) lidocaine and disopyramide preferentially bind to inactivated and activated sodium channels, respectively, because lidocaine's effects are dependent on and disopyramide's effects are independent of APD (during which sodium channels are in the inactivated state); and (2) nicorandil is a useful drug for estimating whether a sodium channel-blocking action of class I antiarrhythmic drugs is due to an inactivated channel block or an activated channel block. These time-dependent reductions of Vmax by both lidocaine and disopyramide were well simulated by the guarded receptor hypothesis. PMID- 2966900 TI - Dementia in Down's syndrome: cerebral glucose utilization, neuropsychological assessment, and neuropathology. AB - We measured the cerebral metabolic rate for glucose (CMRglc) with positron emission tomography and [18F]2-fluoro-2-deoxy-D-glucose in a 47-year-old man with trisomy 21 Down's syndrome (DS) and autopsy-confirmed Alzheimer's disease. Dementia was evident from a confirmed history of cognitive decline, memory loss, and personality change. CMRglc in the subject was compared with the mean obtained in 13 healthy younger DS subjects, aged 19 to 33 years. Test scores of general intelligence, visuospatial ability, language, and memory function showed poorer performance in the older subject compared with the younger group. Mean hemispheric CMRglc in the older DS subject was 28% less than in the young DS group, and marked hypometabolism was evident in parietal and temporal lobe association cortices. At autopsy, extensive neuropathology was noted, especially in the parietal and temporal cortical regions, more so than reported in DS subjects without documented dementia. This study is the first complete assessment of cerebral metabolism, neuropsychological competence, and neuropathology in a DS subject with a documented course of dementia, and demonstrates the superimposition of Alzheimer type dementia on previous mental retardation. PMID- 2966901 TI - Late-onset muscle phosphofructokinase deficiency. AB - A 75-year-old man had a 10-year history of slowly progressive limb weakness without cramps or myoglobinuria. Clinical, morphologic, and biochemical studies showed muscle phosphofructokinase (PFK) deficiency. Erythrocyte PFK activity in his asymptomatic daughter was 63% of normal, compatible with a carrier state. The chronic myopathic variant of muscle PFK deficiency appears to be transmitted as an autosomal recessive trait and may be due to a distinct genetic defect. PMID- 2966902 TI - [Current trends in the diagnostic and therapeutic approach to varicocele]. PMID- 2966903 TI - [Vasculogenic impotence and the Doppler velocimeter examination]. PMID- 2966904 TI - [Use of cyproterone acetate in the treatment of female acne]. PMID- 2966905 TI - Relation between brain dopamine loss and D2 dopamine receptor density in MPTP monkeys. AB - The relationship between dopamine (DA), dihydroxyphenylacetic acid (DOPAC) or homovanillic acid (HVA) concentrations and [3H]spiperone binding to D2 DA receptors in caudate nucleus, putamen and nucleus accumbens following DAergic lesion with MPTP in monkeys was investigated. The level of DA depletion varied from 37% to 100% while a mean elevation in [3H]spiperone binding density of 114.8 +/- 4.7% of control (P less than 0.01) was observed when DA depletion was at least 90% of control values. A logarithmic correlation (y = -7.19 In x +111.15; r = -0.54, P less than 0.01) between DA concentrations (x) and the density of [3H]spiperone binding sites (y) was observed in all brain regions. The correlation between DOPAC or HVA concentrations and [3H]spiperone binding was best represented by linear rather than logarithmic equations. These results indicate that supersensitivity of DA receptors develops after MPTP lesion in monkeys when the number of remaining nerve fibers becomes too few to compensate for the DA loss. PMID- 2966906 TI - Hepatitis B immunisation. PMID- 2966907 TI - Massive ovarian cyst: a comprehensive surgical approach. AB - The massive ovarian cyst (greater than 25 pounds) presents a challenging opportunity for surgical intervention and potential cure of a debilitating problem. Although infrequently seen, such a patient requires awareness of the unique problems removal of these large masses may cause. Successful management requires a comprehensive approach before, during, and after the operative procedure. There are three keys to optimal management: 1) a multisystem, team approach, 2) intact cyst removal, and 3) a special incision and reconstructive technique. PMID- 2966908 TI - A comparison of laparoscopy and laparotomy for the treatment of ectopic pregnancy. AB - A case-control method was used to examine differences in outcome between laparoscopy and laparotomy for the treatment of ectopic pregnancy. Cases and controls, 25 each, were matched by criteria selected to minimize differences in preoperative morbidity and technical difficulty. Patients treated laparoscopically had significantly shorter hospital stays (1.34 +/- 0.8 versus 3.92 +/- 1.1 days; mean +/- SD; P less than .01) and less operating time (77.5 +/ 26.1 versus 103.6 +/- 26.7 minutes; P less than .05). In addition, a laparoscopic approach resulted in a shorter convalescence (8.7 +/- 7.8 versus 25.7 +/- 16.2 days; P less than .01) and reduced postoperative analgesia requirements (0.84 +/- 2.3 versus 4.64 +/- 2.9 doses; P less than .01). We conclude that laparoscopic treatment of ectopic pregnancy is well tolerated by patients and, in comparison with laparotomy, requires fewer medical resources, thereby significantly reducing cost. PMID- 2966909 TI - Work-related therapy for the injured reduces return-to-work barriers. AB - A work hardening program such as the one Mary experienced is designed to assist with case resolution and maximum rehabilitation as expediently and cost effectively as possible. For the worker who cannot successfully return to work due to injury or illness, work hardening is a means of determining what the barriers are to returning to work. As case history showed these barriers often are much more than physical. However, this does not mean that the worker has deep rooted psychological problems. Most of the time, the worker is demonstrating a normal situational reaction to the injury, and once identified, the issues can be resolved expediently. PMID- 2966910 TI - Muscle weakness is leading cause for nurses' lower back injuries, pain. PMID- 2966911 TI - [Cooperative prevention assessed in general practice--a survey of public health offices in North Rhine-Westphalia on the existence of societies for health promotion]. PMID- 2966912 TI - [Health counseling for adults--development of the health counseling office in the Spandau district of Berlin]. PMID- 2966913 TI - [Smoking education campaigns]. PMID- 2966914 TI - [Breast feeding counseling in a baby group--trends in medical education at a social welfare office]. PMID- 2966915 TI - [Behavior disorders of children]. PMID- 2966916 TI - [HIV register of the entire population--a theoretical public health epidemiologic model or a plausible illusion?]. PMID- 2966917 TI - [Treatment and disinfection of swimming and bathing water with reference to Public Health Guideline 19,643]. PMID- 2966918 TI - [Help for AIDS--between social work and organized politics. The article by I. Schafer in Das Offentliche Gesundheitswesen (1987: 389-93) inspires me to comment]. PMID- 2966920 TI - An N-terminal region of adenovirus E1a essential for cell transformation and induction of an epithelial cell growth factor. AB - A new region of the adenovirus E1a protein essential for immortalization and transformation of primary rat kidney cells has been identified. This region is located between amino acid residues 18 to 20 in an N-terminal domain that is not conserved among the various adenovirus serotypes. The transformation defective mutant (18-0) mapping in this region is not impaired in its ability to trans activate the viral E2 promoter and to repress the activity of certain enhancer elements. Mutant 18-0 appears to have only a partial defect in the induction of cellular DNA synthesis in quiescent primary cells suggesting that the N-terminal region plays a role in immortalization and transformation by a mechanism that may not fully depend on induction of cellular DNA synthesis. Mutant 18-0 and another transformation defective mutant (125-7) mapping between amino acid residues 125 to 127 in a conserved domain are defective in the induction of an epithelial cell growth factor, suggesting that growth factor induction may be important for some aspect of adenovirus mediated immortalization and transformation. PMID- 2966919 TI - [ATPase activity and the contractile capacity of the muscle tissue in chick embryos during development]. AB - The value of ATPase activity of the myofibril preparations and the value and duration of actomyosin superprecipitation were estimated for different muscles during the chick embryonic development. The ATPase level increases during embryogenesis 4.5-fold, in the leg muscle this change takes place distinctly earlier than in the leg muscle. The value and rate of actomyosin superprecipitation also markedly increase, to a lesser extent for m. soleus than for m. pectoralis. It is suggested that these changes and differences are mainly due to the delay in synthesis of certain types of the embryonic myosin light chains. PMID- 2966921 TI - Molecular mechanisms of a t(8;14)(q24;q11) translocation juxtaposing c-myc and TcR-alpha genes in a T-cell leukaemia: involvement of a V alpha internal heptamer. AB - We have recently described a t(8;14)(q24;q11) translocation which appeared secondarily in a non-established acute T-cell leukaemia and involved the 3' region of c-myc proto-oncogene and the alpha chain of the T-cell receptor gene (TcR-alpha). In order to elucidate the mechanism of the translocation, we have isolated breakpoint regions from normal and recombinant chromosomes. Our results show that the translocation occurred at the 3' end of a V alpha segment in the proximity of recombination signal sequences (5'-heptamer-23bp spacer-nonamer-3'). Interestingly, an inverted heptamer internal to the V alpha segment was found at two nucleotides 5' of the break. Nucleotide sequence analysis also revealed the presence of homologous signal sequences on chromosome 8, suggesting that the recombination enzymatic system played an important role in the generation of the translocation. This hypothesis is supported by the addition of N nucleotides and the loss of only three nucleotides during the rejoining process. These data established the involvement of a V alpha segment and its recombination signals in the mechanisms of t(8;14) translocations in T-cell leukaemias. PMID- 2966922 TI - Murine c-fms cDNA: cloning, sequence analysis and retroviral expression. AB - We have isolated and sequenced a cDNA clone coding for the murine c-fms gene. The 3677 nucleotide cDNA clone codes for a protein of 976 amino acids, which has 76% and 75% homology to the v-fms and human proteins, respectively. The predicted membrane protein, c-fms, has an amino terminal signal sequence and external, transmembrane and cytoplasmic domains. The homology between murine c-fms and v fms or human c-fms is the strongest (90%-95%) in the cytoplasmic domain and weakest (59%-63%) in the external domain. The c-fms clone was inserted into a retroviral vector containing a neomycin resistance gene and cell lines resistant to G418 were isolated. These cell lines expressed protein which was immunoprecipitated by anti-c-fms antibodies and of the same size as murine c-fms. These cells were also found to specifically bind human CSF-1. PMID- 2966923 TI - Changes in heparan sulfate pattern but not in oncogene expression correlate with tumor growth in spontaneous transformation of cells. AB - For the clonal analysis of spontaneous malignant transformation of cells, a clonal cell line of low tumorigenicity, derived from a mouse embryo mass culture, was injected into syngeneic animals in a sufficient dose (10(7) cells) to produce tumors. Cell lines and clones produced from several such tumors had acquired a 10(4)- to 10(5)-fold higher level of tumorigenicity. Anchorage-independent growth did not co-select with tumorigenicity. There was no evidence for overexpression of proto-oncogenes in the highly tumorigenic clones; p53 protein and mRNA levels were also essentially equal and low. There was a specific structural difference in the O-sulfate residues in oligosaccharides of heparan sulfates in all the tumor cell lines when compared with their parent clone, similar to that observed before in SV40 virus-induced cell transformation (Winterbourne and Mora, 1978). PMID- 2966924 TI - Synthesis, post-translational processing, and autocrine transforming activity of a carboxylterminal truncated form of colony stimulating factor-1. AB - The mononuclear phagocyte colony stimulating factor encoded by a 1.6 kilobase pair human cDNA is synthesized as a homodimeric transmembrane glycoprotein that is released from the plasma membrane by proteolysis. Premature termination of the CSF-1 coding sequence upstream of its carboxylterminal transmembrane-spanning segment and expression of the truncated CSF-1 cDNA in either bovine papilloma virus or retrovirus vectors led to the synthesis of a soluble, biologically active growth factor that was rapidly secreted from cells. Like the full-length CSF-1 precursor, the truncated polypeptide was rapidly assembled through disulfide bonds immediately after synthesis and acquired asparagine-linked oligosaccharide chains that underwent progressive post-translational modifications during intracellular transport. Soluble CSF-1 encoded by the truncated cDNA stimulated the formation of bone marrow-derived mouse macrophage colonies in semisolid medium and induced transformation of mouse NIH-3T3 cells when coexpressed with the human c-fms proto-oncogene product (CSF-1 receptor). Compared to results obtained with the full-length CSF-1 cDNA, the efficiency of transformation obtained with the truncated CSF-1 gene was reduced, in spite of the fact that transfected cultures produced similar levels of the extracellular growth factor. The results indicate that CSF-1 amino acid residues 1-158 (together with the aminoterminal signal peptide at residues -32 to -1) are sufficient for biological activity and that CSF-1 cDNAs encoding either membrane bound or soluble precursors are active in autocrine transformation. PMID- 2966925 TI - Expression of two c-fms-related gene products in rat muscular stem cells. AB - Two c-fms related transcripts 2.0 and 3.7 kb long are continuously expressed during the growth of rat myogenic cells (L6 alpha 1 line) and both are progressively eliminated when cell monolayers reach contact inhibition and differentiate. The 3.7 kb species apparently spans the three domains (intracytoplasmic, transmembrane, extracellular) of the putative c-fms gene(s) while the 2.0 kb species corresponds to the intracytoplasmic domain. In cell-free translation assays, these transcripts directed the synthesis of two proteins of 116 kD and 69 kD respectively. Both transcripts were found to exist in the various lines of rat myoblasts and muscular tumors investigated, and also at low levels in fetal and neonatal muscles. These data and other ones, still preliminary, strongly suggest that two types of c-fms-related gene products have a role in the normal and neoplastic growth of muscular cells. PMID- 2966926 TI - Atlantoaxial instability in individuals with Down syndrome: a fresh look at the evidence. AB - In 1984, the Committee on Sports Medicine of the American Academy of Pediatrics published in this journal a statement on the remarkably high incidence of atlantoaxial instability among individuals with Down syndrome. On the assumption that this instability, demonstrable through a specified series of lateral x-ray films of the neck, constituted a predisposition to cervical spine dislocation with subsequent spinal cord compression, the Academy supported and made more specific a series of recommendations that had originated from the Kennedy Foundation a year previously. In essence, for those persons who are found to have the radiographic sign of instability, participation in sports should be restricted. Because the implementation of these recommendations could deprive tens of thousands of individuals with Down syndrome of activities that are emotionally and physically beneficial and because of the rarity of reported cervical dislocations associated with injury, a case review was done. Included were those cases cited as support for the recommendations along with additional reports that had been omitted and a few cases reported subsequently. Little support for the hypothesis that atlantoaxial "instability" is a predisposing factor to "dislocation" was found, although much was found to indicate an urgent need for carefully designed longitudinal studies. Because nearly all of the cases of actual dislocation were preceded by at least several weeks of readily detectable physical signs, a physical examination with careful attention to neurologic signs prior to participation in sports is more predictive of potential or impending dislocation than the radiologic criteria currently recommended. PMID- 2966927 TI - Atlantoaxial instability and Down syndrome. PMID- 2966928 TI - A new and fast method for preparing high quality lambda DNA suitable for sequencing. AB - A method is described for the rapid purification of high quality lambda DNA. The method can be used from either liquid or plate lysates and on a small scale or a large scale. It relies on the preadsobtion of all polyanions present in the lysate to an "insoluble" anion-exchange matrix (DEAE or TEAE). Phage particles are then disrupted by combined treatment with EDTA/proteinase K and the resulting DNA is precipitated by the addition of the cationic detergent cetyl (or hexadecyl)-trimethyl ammonium bromide-CTAB ("soluble" anion-exchange matrix). The precipitated CTAB-DNA complex is then exchanged to Na-DNA and ethanol precipitated. The resultant purified DNA is suitable for enzymatic reactions and provides a high quality template for dideoxy-sequence analysis. PMID- 2966929 TI - Expression of chromosome 21 specific sequences in normal and Down's syndrome tissues. AB - Using RNA isolated from age and sex matched normal and Down's Syndrome foetal liver and brain tissues, Northern blots were prepared and probed with 4 chromosome 21 specific sequences. The results show that no consistent pattern of expression emerges when Down's Syndrome tissue is compared with normal tissue but the results are very different from the 3/2 ratio of expression which may be expected. Two sequences 21.3 and 26C show only minor differences in expression in trisomy 21 liver samples but significant changes in their expression pattern when normal and Down's Syndrome brain samples are compared. The other sequences, JG77 and JG90 show a 5 fold higher degree of expression in Down's brain but when liver samples are compared one of these sequences shows equal levels of expression in normal and Down's Syndrome samples and the other shows a decrease in expression level in Down's Syndrome samples. PMID- 2966930 TI - The effect of buthobendin on human erythrocyte membrane ATPases. AB - The effect of buthobendin (CravitenR) on the ATPase activities/total (Na, K, Mg) ATPase, and the ouabaine-sensitive (Na, K-ATPase) and insensitive (Mg-ATPase) fractions/from human erythrocyte membrane was examined in the presence or absence of liposomes. The activities of both ATPase fractions increased by 250% in the presence of 10 mmol/l phosphatidylcholine. Buthobendin inhibited both ATPases, but especially Na, K-ATPase. This inhibitory effect was much greater in the hydrophobic milieu of liposomes. The inhibitory effect was proportional to the drug concentration and was stereospecifically related only to the 2S, 2'S isomer. Isomer 2R, 2'R and epinephrine used for comparison did not show any similar inhibitory influence. The decrease in ouabaine-sensitive ATPase activity under the influence of buthobendin might cause changes in cation distribution and cell membrane polarization. An inhibitory effect was also observed in vivo, after single iv administration of buthobendin to patients with ventricular arrhythmias and elevated activities of both erythrocyte membrane ATPases. Complete normalization of cardiac rhythm in about 30% of patients was accompanied - in the sensitive group of patients - with a decrease in Mg-ATPase activity without any changes in elevated Na, K-ATPase activity. In the group of patients resistant to buthobendin therapy both ATPases remained unchanged. PMID- 2966931 TI - Treatment of metastatic prostate carcinoma with the depot LRH analog Zoladex. AB - A long-acting LRH agonist (ICI 118630, Zoladex) was given by monthly subcutaneous injection to 25 patients with previously untreated symptomatic advanced prostatic carcinoma. The medication was well tolerated with the only side effect being hot flushes in 15 patients. Subjective improvement occurred in 22 patients, and disease remission or stabilization judged by objective criteria was seen in 21 and 18 patients from the total group at 3 and 6 months of treatment, respectively. Twelve of 18 patients followed for 1 year were still in objective remission/stabilization. Prostate volume measured by ultrasound decreased by a mean value of 75% and urine flow increased significantly. There were significant falls in serum testosterone and gonadotrophin levels and significant although lesser reductions in serum androstenedione and dehydroepiandrosterone. These changes were accompanied by significant reductions in serum acid and alkaline phosphatase and a rise in serum osteocalcin. Four patients (16%) experienced an initial tumor flare. Although only a small number of patients were studied, Zoladex appeared to be a well-tolerated agent for treatment of prostatic carcinoma, with an initial clinical response similar to that seen with standard endocrine therapy. PMID- 2966933 TI - [Arthroscopy of the hip]. PMID- 2966932 TI - Complications of chiropractic treatment for back pain. AB - Back pain often causes patients great despair, and they expect the primary care physician or orthopedic surgeon to provide a quick, simple solution. Rest and analgesia are the most commonly prescribed treatments, and muscle relaxants, heat, traction, and physiotherapy are also used. If these treatments do not help, the patient may search for relief through faith healing, acupuncture, chiropractic treatment, or other nonconventional forms of treatment. Although chiropractic treatment is a popular alternative, its long-term effect is questionable and the medical literature contains numerous reports of patients whose condition worsened as a result of it. Physicians should be aware of the dangers of chiropractic treatment, particularly in patients with severe spondylitic changes, osteoporosis, fractures, tumors, ankylosing spondylitis, infections, or signs of nerve root pressure. PMID- 2966935 TI - [Etiological diagnosis of syncope and brief loss of consciousness in patients over 65. Results of a multicenter prospective study of 188 cases]. AB - In a prospective multicentre study a series of 188 patients over 65 years of age admitted for syncope (90) or transient loss of consciousness (98) was collected. There were 120 women and 68 men; mean age was 78.7 +/- 6.6 years; 140 patients had a significant history mainly of arterial hypertension and coronary disease; 81 patients were taking drugs (4.1 on average) and notably diuretics, antihypertensive agents, antiarrhythmic agents, nitrites and psychotropic drugs. The most frequent diagnoses were arrhythmia (20.2 p. 100), postural hypotension (15.4 p. 100), reflex and vasodepressive syncope (12.2 p. 100) and coronary disease (5.3 p. 100). Taken together, these diagnoses could be divided into cardiac diseases (26.5 p. 100), extracardiac diseases (54.2 p. 100), diagnosis unknown (19 p. 100). The condition was iatrogenic in 45 patients (24 p. 100). The means by which the diagnosis was reached were studied: in two-thirds of the cases, careful physical examination and electrocardiography alone provided a diagnosis. Seven patients died during their stay in hospital. Among the 181 survivors 148 (81.7 p. 100) were discharged and returned home. PMID- 2966934 TI - [Prevention of hemorrhagic recurrence in cirrhotic patients. Is sclerotherapy better than propranolol?]. AB - The effectiveness of oral propranolol and sclerotherapy in preventing recurrent bleeding after an endoscopically proven haemorrhage from oesophageal varices was compared in Pugh's grade B and C patients divided into two successive therapeutic groups. Group I patients (n = 32) were given oral propranolol, while sclerotherapy was performed in group II patients (n = 32), 23 of whom simultaneously received propranolol. There was no difference between groups I and II in the severity of the initial bleeding and liver failure (Groups I/II, B 19/18, C 13/14), in the withdrawal of alcohol (group I 25, group II 17) and in compliance with either treatment (group I 25, group II 27). Obliteration of the oesophageal varices was achieved in 69 p. 100 of group II patients within a median number of 4 courses and 46 days, and in 84 p. 100 of the patients of this group who survived for more than 3 months. One year after the beginning of treatment rebleeding had occurred in 43 p. 100 of either group I or group II patients. The 1-year survival rate was 72 p. 100 in group I and 62 p. 100 in group II (non significant). The causes of death were the same in both groups. Irrespective of the treatment assigned, death was 2.1 times more frequent (P = 0.02) and rebleeding was 1.2 time more frequent (P = 0.08) in grade C than in grade B patients. PMID- 2966936 TI - [1-year prognosis of syncope and brief loss of consciousness in patients over 65. A multicenter study of 188 cases]. AB - The one-year outcome of syncope and transient loss of consciousness was studied prospectively in 188 patients aged over 65 admitted to internal medicine departments. Thirty-seven patients (19.6 p. 100) died. This mortality rate at one year was 2.34 times higher than that of a non-hospitalized french population of the same age group (standardized mortality ratio, SMR = 2.34, P less than 0.001). The frequency of sudden death (7 cases) was much higher than that observed in the reference population (0.05 expected deaths, P less than 0.0001). Overmortality was clearly apparent in groups with an initial diagnosis of heart disease (SMR = 2.36, P less than 0.01) or neurological disease (SMR = 4.25, P less than 0.001). The relapse rate was 28 p. 100 globally and up to 43 p. 100 in cardiac patients. In the group with iatrogenic symptoms treatment was appropriately corrected in 86 p. 100 of the patients, and none of these relapsed. One year after the initial episode 11 p. 100 of the surviving patients had been institutionalized. PMID- 2966938 TI - [Cowden's disease]. PMID- 2966937 TI - [Pharmacokinetics of prednisolone in children. Study of a correlation with tolerability and therapeutic effect in nephrosis]. AB - In patients with nephrotic syndrome the response to corticosteroids and the way these drugs are tolerated are extremely variable. The purpose of this study was to investigate possible correlations between the pharmacokinetic values of prednisolone and the main clinical criteria of effectiveness and safety. The study was performed on 18 children under corticosteroid therapy: 16 with nephrotic syndrome and 2 with a systemic disease. Measurements were performed by radiocompetition with transcortine after an oral dose of 1 mg/kg bodyweight. Pharmacokinetic values varied considerably, with peak plasma levels ranging from 1.2 to 6.1 micrograms/ml between 20 and 120 minutes, and T 1/2 values of 77 to 648 minutes. Within this scattering of values, some patients were clearly outside the mean T 1/2 value (3.1 +/- 1 hours) due to a particularly fast or slow metabolisation of the drug. In patients with nephrotic syndrome no correlation was found between pharmacokinetic values and criteria of clinical effectiveness, such as the time and dosage required to obtain remissions and the duration of these remissions. Hypoalbuminaemia had no influence on the metabolism of prednisolone. In contrast, there was a correlation between pharmacokinetic values and side-effects, since patients who presented with side-effects also had a significantly greater area under plasma concentration versus time curves. This pharmacokinetic test may be used when corticosteroids produce unusual or unexplainable therapeutic results or adverse reactions. PMID- 2966939 TI - [Ovarian localization disclosing lymphoma a posteriori]. PMID- 2966940 TI - [Pemphigus induced by alpha-mercaptopropionylglycine]. PMID- 2966941 TI - [Pseudoalcoholic hepatitis during treatment with nicardipine]. PMID- 2966942 TI - [Surgical excision of intramyocardial cirsoid aneurysm]. PMID- 2966943 TI - [Isolation of Mycobacterium tuberculosis hominis from hemocultures in acquired immunodeficiency syndrome]. PMID- 2966944 TI - [Legionnaires' disease with agranulocytosis of favorable course]. PMID- 2966945 TI - [Rheumatoid purpura following influenza vaccination]. PMID- 2966946 TI - [Study of changes in behavior of Parisian intravenous drug addicts]. PMID- 2966947 TI - [Study of the central motor pathway by electromagnetic cortico-spinal cord stimulation]. PMID- 2966948 TI - [Precursors of polypeptide hormones: the model of proopiomelanocortin. From the molecular puzzle to regulation of gene expression]. PMID- 2966949 TI - [Dietetics of mucoviscidosis. Toward a normolipid diet]. PMID- 2966950 TI - [Value of postoperative radiotherapy in various tumors of the pituitary gland]. AB - Already advocated by Cushing to complement surgery of pituitary adenomas, radiotherapy remains useful in these benign tumours despite controversies concerning long-term results, pituitary cell resistance and potential harmful effects on the optic nerve, glands and skin. Radiotherapy was applied to 36.6 p. 100 of 150 patients with pituitary tumours operated upon by the same surgical team. Some very good results were obtained, notably in invasive and recurrent tumours. In a case of chromophobe adenoma operated twice at 1 year interval, radiotherapy (4,500 rads) after the second operation resulted in complete cure confirmed by computerized tomography 15 years later. Similarly, in an 8-year old girl reoperated upon for severe recurrent suprasellar craniopharyngioma 3 years after the initial excision, post-operative radiotherapy (5,000 rads) produced excellent remission which persists after 19 years. Thus, even with the modern diagnostic and microneurosurgical procedures, radiotherapy remains, for many authors, a useful and safe method to improve the prognosis of pituitary tumours. PMID- 2966951 TI - [Joint complications in patients with chronic renal failure hemodialyzed for over 10 years. 40 cases]. AB - A retrospective study of 40 patients with chronic renal failure who underwent haemodialysis for more than 10 years (mean: 153 months) showed that 18 patients (45 p. 100) had arthralgia in the shoulders, hands, wrists and knees, 13 (32 p. 100) had carpal tunnel syndrome requiring surgery, and 20 (50 p. 100) were found to have bone cavities in the humeral head, external supra-acetabular region, carpus and patella. Aluminium overload was present in 47 p. 100 of the patients, and amyloid deposits were found in 10 of the 12 patient operated upon for carpal tunnel syndrome. This study confirms the frequency in patients under long-term haemodialysis of an articular pathological entity consisting of arthralgia, carpal tunnel syndrome, juxta-articular bone cavities and amyloid deposits which are now known to be made of beta 2-microglobulin. The initial lesion seems to affect the synovial membrane; it appears to be facilitated by age and is often associated with aluminium overload. The mechanism(s) responsible for amyloid deposits remain (s) to be elucidated. PMID- 2966952 TI - [Reduction of postprandial gastroesophageal reflux with sodium alginate in suspension. Multicenter pH study in 21 patients]. AB - The purpose of this study was to ascertain the pharmacological effectiveness of sodium alginate liquid in the treatment of gastroesophageal acid reflux. The pharmacological test selected was 3 hours postprandial oesophageal pH, because of the physico-chemical properties of this drug. Twenty-one patients of both sexes were included in this study. The initial pH measurement served both as inclusion criteria, by confirming the presence of gastric acid reflux, and as reference for the evaluation of the effect of sodium alginate liquid by a second pH determination after treatment. After treatment with sodium alginate liquid, the number and mean duration of episodes of reflux, as well as the time spent at each pH level, had significantly decreased as compared to the initial value. The gastroesophageal reflux score was significantly reduced during the second hour in recumbent posture, but none of the different parameters studied separately (number and mean duration of episodes of reflux and time spent at each pH level) was significantly decreased. This study demonstrates the pharmacological effectiveness of the drug during the post prandial period and indicates that it is more effective in upright than in supine position. PMID- 2966953 TI - [Hypoglycemia caused by overdose of a new anti-arrhythmia agent: cibenzoline. 3 cases]. AB - Severe fasting hypoglycaemia were observed in three patients treated with cibenzoline, a new class I antiarrhythmic drug. Retrospective analysis of electrocardiographic changes and serum level determinations showed that these reactions were contemporaneous with intoxication by overdosage of the drug. None of the patients was taking any other drug known to produce this adverse reaction, and in the absence of any other cause of hypoglycaemia, we concluded that it was related to the administration of cibenzoline. Thus, cibenzoline is yet another class I antiarrhythmic agent producing hypoglycaemia as an adverse reaction. Serum insulin levels measured in two of the three patients during hypoglycaemia were inappropriately high, suggesting that cibenzoline might stimulate insulin secretion. PMID- 2966955 TI - [Recurrent pneumococcal meningitis associated with C3 deficiency]. PMID- 2966954 TI - [The role of the platelet-activating factor (PAF-acether) in renal pathology]. AB - Platelet activating factor (PAF-acether) is emerging as a potentially crucial mediator of kidney physiopathology. Several experimental studies indicate that the mediator is implicated, directly or via other agonists, in several renal diseases. PMID- 2966956 TI - [Perforations of the small intestine caused by tablets of potassium chloride]. PMID- 2966957 TI - [Meningoradiculoencephalitis in Lyme disease. A case with major regressive mental disorders]. PMID- 2966958 TI - [Relations between sleep stages and asthma crisis: continuous study of electroencephalography and capnography]. PMID- 2966959 TI - [Polymorphism of DNA in renal polycystosis]. PMID- 2966960 TI - [HLA-B27 inflammatory spondylarthropathy, psoriasis and HIV infection]. PMID- 2966961 TI - Atrial natriuretic factors: a review. PMID- 2966962 TI - Mutation of aspartic acid-351, lysine-352, and lysine-515 alters the Ca2+ transport activity of the Ca2+-ATPase expressed in COS-1 cells. AB - Full-length cDNAs encoding neonatal and adult isoforms of the Ca2+-ATPase of rabbit fast-twitch skeletal muscle sarcoplasmic reticulum were expressed transiently in COS-1 cells. The microsomal fraction isolated from transfected COS 1 cells contained immunoreactive Ca2+-ATPase and catalyzed Ca2+ transport at rates at least 15-fold above controls. No differences were observed in either the rates or Ca2+ dependency of Ca2+ transport catalyzed by the two isoforms. Aspartic acid-351, the site of formation of the catalytic acyl phosphate in the enzyme, was mutated to asparagine, glutamic acid, serine, threonine, histidine, or alanine. In every case, Ca2+ transport activity and Ca2+-dependent phosphorylation were eliminated. Ca2+ transport was also eliminated by mutation of lysine-352 to arginine, glutamine, or glutamic acid or by mutation of Asp351 Lys352 to Lys351-Asp352. Mutation of lysine-515, the site of fluorescein isothiocyanate modification in the enzyme, resulted in diminished Ca2+ transport activity as follows: arginine, 60%; glutamine, 25%; glutamic acid, 5%. These results demonstrate the absolute requirement of acylphosphate formation for the Ca2+ transport function and define a residue important for ATP binding. They also demonstrate the feasibility of a thorough analysis of active sites in the Ca2+ ATPase by expression and site-specific mutagenesis. PMID- 2966963 TI - Hypothyroidism and pituitary contents of immunoactive met-enkephalin and beta endorphin in male rats of different ages. AB - A comparison of the effect of PTU-induced hypothyroidism on the contents of immunoactive met-enkephalin and beta-endorphin in the pituitaries from rats aged 2 1/2, 12, and 18 months was made. In all ages, there was significant reduction of IR-met-enkephalin content in the anterior lobe and IR-beta-endorphin content in the neuro-intermediate lobe after PTU treatment. There was a significant age related decrease in IR-beta-endorphin content in the anterior lobe. Rats of all three ages responded to PTU treatment with an increase in serum TSH level and a drastic reduction in serum T3 and T4 levels. The results indicate that there was no age-related difference in the change of pituitary opioid peptide contents in response to hypothyroidism. PMID- 2966964 TI - Adrenocortical, beta-endorphin and behavioral responses to graded stressors in differentially reared rats. AB - Isolation rearing has long been suspected to alter hormonal and behavioral responses to stress. Two experiments were conducted to test the hypothesis that isolates are more timid or fearful than socially reared rats when exposed to novel test environments. In both, isolate response to 3 graded stressors was compared to that of socially-reared rats. In the first experiment, animals were handled, shocked or not treated prior to testing to produce three levels of conditioned fear. They were then tested on four paradigms previously shown sensitive to conditioned fear: open field activity, emergence latency, auditory startle, and latency to accept food from the experimenter. In the second experiment, rats were given a 0-, 5- or 20-min forced swim, then sacrificed for analysis of plasma corticosterone and pituitary and hypothalamic beta-endorphin. It was found that isolates showed little evidence of enhanced behavioral timidity, although rearing effects were seen on all 4 behavioral measures. Plasma corticosterone levels increased in a graded fashion over the course of the forced swim, but there was no effect of rearing conditions. While there were no effects of rearing or stress on hypothalamic beta-endorphin, pituitary beta-endorphin content was lower in females than in males, and isolate males had lower pituitary endorphin than social males. In summary, these experiments provide no evidence that isolation rearing produces a primary, global increase in fearfulness, but identify several behavioral and hormonal differences associated with differential housing in rats. PMID- 2966965 TI - Duration of effect of oral diphosphonate therapy in Paget's disease of bone. AB - The effects of the diphosphonates etidronate and clodronate were studied in 144 patients with Paget's disease. All five programmes of treatment tested induced a similar suppression of disease activity as judged by serum alkaline phosphatase concentrations, but the proportion of patients responding and the duration of responses differed significantly between programmes. The proportion responding to etidronate 5-10 mg/kg/day for six months was less than for other regimens, and the most sustained response was after treatment with clodronate 1600 mg daily for six months. More complete biochemical suppression was associated with the more prolonged responses irrespective of the regimen used. PMID- 2966966 TI - A painful adolescent back. PMID- 2966967 TI - Legal case briefs for nurses. MT.: earning capacity: workers' compensation; R.I.: change in codition: workers' compensation. PMID- 2966968 TI - Effects of hypertrophy on the coronary circulation. PMID- 2966969 TI - Thromboxane receptors can modulate gastric acid secretion in the rat. AB - The effects of PGE2 and the thromboxane A2 mimetic, U-46619, have been investigated on gastric secretion in the rat isolated gastric mucosa. Both compounds produced concentration-related inhibitions of histamine-induced secretion whereas only U-46619 inhibited methacholine-stimulated and basal secretion, and neither compound had any effect on the secretory response to dbcAMP. Indomethacin had no effect on the antisecretory activity of PGE2 but markedly reduced the potency of U-46619 suggesting that endogenous prostaglandins play a role in the U-46619 responses. However, direct inhibitory effects of U 46619 were seen at high concentrations. The thromboxane receptor antagonist AH23848, at concentrations selective for thromboxane receptors, had no effect on responses to PGE2 but markedly inhibited the effects of U-46619. We conclude that the antisecretory profile of U-46619 differs from that of PGE2. U-46619 has both direct and indirect antisecretory effects and these are mediated via thromboxane receptors in the rat gastric mucosa. PMID- 2966970 TI - [Tinea pedis in workers in the Lubiana dinnerware porcelain factory in Lubiana near Koscierzyna]. PMID- 2966971 TI - [Benzoyl superoxide in the treatment of acne vulgaris]. PMID- 2966972 TI - [Treatment of acne vulgaris by local use of erythromycin]. PMID- 2966974 TI - [Diagnosis of right ventricular overloading by means of the magnetocardiography]. PMID- 2966973 TI - [Infectious endocarditis in hypertrophic myocardiopathy: problems of management]. PMID- 2966975 TI - [Glucocorticoids and lipocortin]. PMID- 2966976 TI - [Benign intracranial hypertension and minocycline]. AB - A 19 year-old woman complained of headache and nausea occurring while she was taking minocycline for acne. Examination showed bilateral papilloedema and a bilateral VIth nerve palsy. Symptoms and signs rapidly resolved after the drug was stopped. Benign intracranial hypertension due to tetracyclines is well known in infants. It is rare in adults. Its pathophysiology remains unknown. The role of vitamin A is inconsistent. Others biological factors or personal susceptibility could be involved. PMID- 2966977 TI - Rhinophyma: treatment by excision and silver impregnated amniotic membrane. PMID- 2966978 TI - [Alloplastic replacement of the anterior cruciate ligament. Histomorphologic studies of ruptured Dacron ligaments]. PMID- 2966979 TI - Progesterone therapy for malignant peritoneal cytology surgical stage I endometrial adenocarcinoma. AB - Twenty-five patients with surgical stage I endometrial adenocarcinoma and malignant peritoneal cytologic washings were treated with progesterone therapy. Twenty-two patients have undergone a second-look laparoscopy and repeat cytologic washings, one of whom also underwent a third-look laparoscopy. Two patients refused second-look laparoscopy, and in a third patient, a laparoscopy was medically contraindicated; the three patients have no evidence of disease at 15, 46, and 64 months, respectively, and are off therapy. Of the 22 patients who underwent second-look laparoscopy, 21 (95%) had no evidence of recurrent endometrial carcinoma and repeat peritoneal results of cytology were negative for malignant cells; one patient (5%) had persistent malignant peritoneal cytology at second-look laparoscopy but had no evidence of disease at third-look laparoscopy after an additional year of progesterone therapy. All 25 patients (100%) are off progesterone therapy and remain clinically free of disease from 12 to 64 months after discontinuation of therapy. PMID- 2966980 TI - Preclinical studies with trimetrexate: a review of conclusions and unanswered questions. PMID- 2966981 TI - Safety and tolerance of trimetrexate: results of a phase II multicenter study in patients with metastatic cancer refractory to conventional therapy or for which no conventional therapy exists. PMID- 2966982 TI - Trimetrexate as a single agent in patients with advanced head and neck cancer. AB - Trimetrexate is a nonclassical folate antagonist. It was evaluated in 43 patients, previously not exposed to chemotherapy, with incurable and/or demonstrated progression of squamous cell head and neck cancer following surgery and/or radiotherapy. Trimetrexate was initially administered at 8 mg/m2 intravenous (IV) daily for 5 days, repeated every 3 to 4 weeks with dose adjustments depending on patient tolerance. Thirty-eight of the 43 patients entered in this study were evaluable for response. The 28 males and 10 females had a median age of 60 years. Their median performance status on the Zubrod scale was 1. Thirty-seven patients (97%) had received prior radiotherapy. The disease had metastasized in 84% of the patients with dissemination primarily to the subcutaneous soft tissue (40%). Ten patients (26%; with a 95% confidence interval of 12% to 40%) achieved a partial response (PR) with a median duration of 12.2 weeks (median time to PR was 3.2 weeks). The median survival of the 43 patients was 17.3 weeks with 10 patients alive at last follow-up. Hematologic grade 3 to 4 toxicities included leukopenia (19%), thrombocytopenia (9%), and granulocytopenia (5%). Nonhematologic grade 3 to 4 toxicities possibly attributable to trimetrexate treatment included stomatitis (7%). PMID- 2966983 TI - Trimetrexate: a phase 2 study in previously treated patients with metastatic breast cancer. AB - Trimetrexate, a nonclassical antifolate, was administered to 28 patients with progressive, metastatic breast cancer. The starting dose was 8 mg/m2 administered intravenously (IV) as a short infusion daily for 5 days, repeated every 3 weeks with dose adjustments depending on patient tolerance. The median age of the 28 females in this study was 58.5 years. Their median performance status on the Zubrod scale was 1. Nine of the patients were estrogen-receptor positive. The sites of metastasis included the pulmonary system (50%), bone (40%), and liver (35%). Twenty patients were evaluable for response. Three patients (15%; [95% confidence interval 0.0, 30.6]) achieved a partial response with a median time to partial response of 9.1 weeks and a median duration of partial response of 13.1 weeks. Two of the responders were estrogen-receptor positive. Median survival for all 28 patients was 41 + weeks with 15 patients alive at the last follow-up. Hematologic grade 3-4 toxicity occurred as follows: leukopenia (21%), thrombocytopenia (18%), and granulocytopenia (18%). There was no significant nonhematologic toxicity seen. PMID- 2966984 TI - Phase 1 and 2 studies of trimetrexate administered in combination with fluorouracil to patients with metastatic cancer. PMID- 2966985 TI - Treatment of Pneumocystis carinii pneumonia with trimetrexate in acquired immunodeficiency syndrome (AIDS). AB - In vitro studies have shown that trimetrexate, a lipid-soluble analogue of methotrexate, is 1500 times more potent than trimethoprim as an inhibitor of dihydrofolate reductase from Pneumocystis carinii. Furthermore, trimetrexate is readily taken up by P carinii, while performed folates such as leucovorin are not. These observations suggest that the combination of trimetrexate plus leucovorin, which can specifically protect mammalian host tissues from the toxic effects of the antifolate, may be useful in the treatment of pneumocystis pneumonia. This concept was tested in a clinical study of 49 patients with acquired immunodeficiency syndrome (AIDS) and P carinii pneumonia who were treated for 21 days with trimetrexate and leucovorin. Patients were divided into three groups: 16 patients who were unable to tolerate or had failed both pentamidine isethionate and trimethoprim-sulfamethoxazole therapy were treated with trimetrexate plus leucovorin (Group I); 16 patients who were unable to tolerate sulfonamide therapy were treated with trimetrexate with leucovorin as initial therapy (Group II); and 17 patients in whom trimetrexate with leucovorin plus sulfadiazine was used as initial therapy (Group III). Response and survival rates were 69% and 69% in Group I; 63% and 88%, respectively, in Group II; and 71% and 76%, respectively, in Group III. Toxicity was minimal. The results indicate that trimetrexate with leucovorin is safe and effective for initial therapy in AIDS patients with P carinii pneumonia and in those intolerant or unresponsive to standard therapies. PMID- 2966986 TI - Trimetrexate: overall clinical results. PMID- 2966987 TI - Clinical pharmacology and metabolism of trimetrexate. PMID- 2966988 TI - Thrombosis of the superior mesenteric vein in a patient with recurrent spontaneous venous thrombosis caused by familial protein S deficiency. PMID- 2966989 TI - [Characteristics of intracardiac hemodynamic disorders in patients with dilated cardiomyopathy]. PMID- 2966990 TI - [Current methods of follow-up and correction of the performance of the physical rehabilitation of patients with ischemic heart disease at sanatoria of the Caucasian Mineral Waters health resort]. AB - A total of 1055 CHD patients aged 45 to 70 (680 men and 375 women) were examined during physical rehabilitation using tele-electrocardiographic and monitor control in the Caucasian Mineral Waters health resorts. A retrospective analysis of 400 case histories of CHD patients treated in the same sanatoria in 1985 without the use of present-day methods of control over physical training, was taken as control. Short- and long-term results showed that therapeutic efficacy was higher in the group where the levels of physical exercises were corrected by regular tele-electrocardiographic and monitor control. PMID- 2966991 TI - [Analysis of different lymphocyte subpopulations in patients with nonspecific ulcerative colitis]. AB - A complex of methods based on a rosette forming reaction, was used to study the content of different lymphocytic populations in the peripheral blood of persons with nonspecific ulcerative colitis. Imbalance in the system of immunocompetent cells was shown to develop in this disease: a decrease in the amount of suppressors (T gamma and theophylline sensitive lymphocytes) and a increase in the amount of immature postthymic cells: cells forming rosettes with autologous erythrocytes, and theophylline dependent lymphocytes. PMID- 2966992 TI - [Effectiveness of long-term treatment of patients with hypertension]. AB - Dispensary care was provided to 122 patients with essential hypertension. During a 3-year follow-up period BP decrease in regularly treated patients (44) was accompanied by the restoration of hemodynamic cardiac function and regression of myocardial hypertrophy and ischemia. BP stabilization, transformation of the blood circulation of hemodynamic type, an increased degree of myocardial hypertrophy and ischemia were noted in irregularly treated patients. PMID- 2966993 TI - Genetics of susceptibility to 6-aminonicotinamide-induced cleft palate in the mouse: studies in congenic and recombinant inbred strains. AB - In a search for genetic differences in susceptibility to cleft palate, congenic and recombinant inbred strains of mice were treated with 6-aminonicotinamide or control injections. Of six loci tested, only the chromosome segment marked by N acetyl transferase was found to affect susceptibility to 6-aminonicotinamide induced cleft palate. This chromosome segment is known to affect glucocorticoid induced cleft palate and phenytoin-induced cleft lip with or without cleft palate in these strains of mice. PMID- 2966994 TI - Observations with anteovin in dermatological disorders. PMID- 2966995 TI - Comparison of benefit against risk of ketoconazole therapy. PMID- 2966996 TI - [Disease and depression of productivity in raising swine caused by mycotoxins]. AB - Mycotoxins are natural substances, which are poisonous for man and animals. Mycotoxicological investigations, carried out from 1982 to 1986, showed that aflatoxins (B1, B2, G1, G2), trichothecenes (T-2 toxin, HT-2 toxin, diacetoxyscirpenol, monoacetoxyscirpenol, neosolaniol and deoxynivalenol) ochratoxin A and zearalenone are to be expected in feedstuffs. Further, sclerotia of Claviceps purpurea were found in cereals repeatedly, which demonstrates that also ergot alkaloids can be found in feedstuffs. The determined toxin concentrations often do not suffice to produce the described "classic" clinical symptoms and pathological changes. On the contrary the continuous intake of small amounts leads to chronic intoxications which are characterized by loss of weight, insufficient gain in weight, fertility disorder or increased susceptibility for infectious diseases. As feedstuffs are mostly contaminated by different kinds of toxinogenic fungi and as fungal strains are frequently able to produce several mycotoxins simultaneously, it must be assumed that naturally occurring mycotoxicoses are not monocausal, but multitoxic diseases. Although we have improved analytical methods, naturally occurring mycotoxicoses still cannot always be cleared up completely. PMID- 2966997 TI - Double-blind study of traveller's diarrhoea using Nifuroxazide. PMID- 2966998 TI - Beneficial effect of atrial natriuretic factor on ischemically injured kidneys in the rat. A new approach to improve early renal function. AB - In a rat experimental study we investigated whether the atrial natriuretic peptide by itself is able to improve early renal function after an ischemic injury. Two groups of Wistar male rats underwent a right nephrectomy and a left renal artery occlusion for 30 min and were infused for 2 hr after ischemia with isotonic saline or rat atrial natriuretic peptides (alpha ANF: 28 amino acids (AP 28) and atriopeptin III (AP 24): 24 amino acids). ANF infusion increased the urinary flow (P less than 0.001), the urinary sodium concentration (P less than 0.001), the sodium excretion rate (P less than 0.0001), and improved the glomerular filtration rate (GFR) recovery (P less than 0.02) determined at the end of the 2-hr infusion period. AP 24 exhibited higher natriuretics activities than AP 28. The effect of both peptides upon GFR recovery was equivalent. These effects of ANF observed after acute ischemia suggest that this peptide may be beneficial on the resumption of renal function in the early phases following transplantation. PMID- 2966999 TI - Characterization of peripheral blood CD8/11 cells in bone marrow transplant recipients. II. Two distinct populations of CD8/11 cells. AB - Flow cytometric analysis of mononuclear cell surface antigens identified 2 distinct populations of CD8/11 cells in the peripheral blood of recovering bone marrow transplant (BMT) recipients. These populations were distinguished based on differential CD8 antigen expression. One population expressed high-surface density CD8 (CD8 bright/11); the other population expressed low-surface density CD8 (CD8 dim/11). The surface expression of CD11, the C3bi receptor on lymphocytes, is similar in CD8 right/11 and CD8 dim cells. Thus, although 13 BMT recipients had elevated CD8/11 cells (mean = 27% +/- 9%; normal range mean = 7.4 +/- 3.3%), the percent of CD8 bright/11 cells versus CD8 dim/11 cells varied. The majority of cells in patients with a predominant CD8 right/11 phenotype did not express CD16 (Fc gamma receptor). In contrast, the CD8 dim/11 cells coexpressed CD16. When functional studies were performed, we observed high numbers of CD8 right/11 cells correlated with low natural killer (NK) lytic activity, while patients with less than 25% CD bright/11 cells had high NK activity. Analysis of interleukin 2 (IL-2) production revealed that none of the peripheral blood mononuclear cells (PBMC) from BMT patients synthesized IL-2 at less than 1.5 months posttransplant. However, cells from some patients began to synthesize IL-2 at approximately 2 months posttransplant. It is likely that both populations of CD8/11 cells have an impact on the regeneration and regulation of the immune system in BMT recipients. PMID- 2967000 TI - Effect of blood transfusion on IL-2 production. AB - Spleen cells from B10.A mice transfused with B10.D2 blood suppress the immune responses of normal B10.A to B10.D2 in coculture as early as 2 days posttransfusion. In addition, the ability of B10.A mice to respond in cell mediated lymphocytotoxicity (CML) is significantly impaired as early as 2 days after B10.D2 transfusion. Experiments were performed to characterize the cells mediating the suppressive effect and to determine whether the inability of transfused mice to generate a cytotoxic response is due to an inhibition of IL-2 production. To characterize the suppressor cells, spleen cells from B10.A mice were assayed 2 or 16 days after B10.D2 transfusion for the ability to suppress mixed lymphocyte culture (MLC) and CML responses of normal B10.A mice in coculture. The putative suppressor cells were either passed over a Sephadex G-10 or nylon wool column, treated with anti-Thy antibody or left untreated before addition to the coculture. Untreated cells from transfused mice suppressed the CML response of normal B10.A both 2 and 16 days posttransfusion, while the effect on the MLC response was inconsistent. Passage of the cells over Sephadex G-10 or nylon wool before assaying abrogated the suppressive effect, while treatment with anti-Thy antibody had no effect. These results suggest that the suppressor cells appearing shortly after blood transfusion have the characteristics of macrophages and not T lymphocytes. To determine the effect of transfusion on IL-2 production, cells from transfused mice were assayed for their ability to produce IL-1 and IL 2 and for the formation of IL-2 receptors. In addition, the effect of exogenous IL-1 and IL-2 on restoring the CML response of transfused mice to normal was assayed. The production of IL-1 by transfused mice was normal, while the production of IL-2 was significantly suppressed both 2 and 16 days posttransfusion. Activated cells from normal and transfused mice showed equal ability to absorb IL-2, indicating that IL-2 receptor formation is normal after transfusion. The addition of exogenous IL-2, but not IL-1, to CML cultures containing cells from transfused mice as responders restored the response to normal. These results indicate that the inability of transfused mice to respond in CML is due, at least in part, to an inability to produce IL-2. This could be mediated by prostaglandins released by activated macrophages. PMID- 2967001 TI - Enhancement of tumor growth by suppressor factors derived from a natural suppressor/cytotoxic cell line. PMID- 2967002 TI - [Multihandicapped children in the region of Copenhagen. Evaluation of occurrence and of the pattern of referral of children with extensive mental and physical handicaps in the County of Copenhagen and the Municipalities of Copenhagen and Fredriksberg 1970-1985]. PMID- 2967003 TI - [Occurrence and survival of newborn infants with Down syndrome in Denmark 1980 1985]. PMID- 2967004 TI - [Itching in a factory]. PMID- 2967005 TI - [Antabuse and nickel allergy]. PMID- 2967006 TI - [Measurement of blood circulation by Doppler ultrasound]. PMID- 2967007 TI - [The course of a vaccination program for hospital staff]. PMID- 2967008 TI - Low back pain disorders. PMID- 2967009 TI - Angioplasty: long-term experience of the Mayo Clinic. PMID- 2967010 TI - [Causes of eye enucleation during the 10-year period in the Mordov ASSR]. PMID- 2967012 TI - Virus-lymphocyte interactions. III. Biologic parameters of a virus variant that fails to generate CTL and establishes persistent infection in immunocompetent hosts. AB - Viruses that cause in vivo persistent infections avoid the host's immunologic surveillance machinery. A major component of that armamentarium is virus-specific MHC-restricted cytotoxic T lymphocyte (CTL) response of the host. Studies with lymphocytic choriomeningitis virus (LCMV) have uncovered a parental virus (CTL+) that in immuno-competent adults induces CTL and terminates acute infection and a variant (CTL-) that fails to elicit CTL responses and establishes a persistent state (R. Ahmed et al. (1984) J. Exp. Med. 160, 521-540). The biologic properties, similarities, and differences between CTL+ and CTL- viruses as regards their interactions with lymphocytes of newborn and adult mice is recorded here. CTL+ and CTL- viruses persist in lymphocytes of newborn inoculated mice, primarily within the T helper subset. Approximately 2% of lymphocytes express viral nucleic acid sequences while only 0.04% score as infectious centers suggesting incomplete viral replication. These levels were maintained over the course of infectious. In contrast, CTL- virus but not CTL+ persists in lymphocytes of mice inoculated when adults. Lymphocytes easily scored as infecting centers but rarely displayed nucleic acid sequences suggesting a different balance of incomplete to complete virion replication. Further, infectious centers decreased by 10-fold from the 3rd to 68th day of infection and the total numbers of T lymphocytes in the circulation decreased suggesting CTL- may replicate in and destroy lymphocytes of adult mice. In the following paper the primary nucleotide structure of the LCMV small RNA segment, the segment responsible for generation of CTL and encoding the proteins recognized by CTL, for CTL+ and CTL- viruses is reported. PMID- 2967011 TI - Antibody development and cellular immune responses in sheep immunized and challenged with Sarcocystis tenella sporocysts. AB - Four specific-pathogen-free (SPF) sheep were experimentally infected with 10(3) or 10(4) Sarcocystis tenella (syn. S. ovicanis) sporocysts and another two sheep served as uninfected controls. All sheep were challenged 49 days later by infection with 2.5 X 10(5) sporocysts and their humoral and cellular responses to infection and challenge were assessed weekly by enzyme immunoassays and lymphocyte transformation assays. The control sheep died from acute sarcocystosis 29-30 days after challenge, whereas the immunized sheep survived and were protected against acute disease. Specific IgM and IgG antibodies were detected in the immunized sheep from 28 days after infection onwards. Lymphocytes collected before and after challenge did not exhibit any significant differences in their responses to stimulation with S. tenella cystozoite or sporozoite antigens. Furthermore, lymphocytes collected before challenge did not differ from the controls in their responses to stimulation with the mitogens lipopolysaccharide or phytohaemagglutinin. However, lymphocytes collected after challenge did exhibit increased blastogenic responses to stimulation with both mitogens from 21 28 days after challenge onwards. The infected sheep were necropsied 46 days after challenge, and histological and ultrastructural studies revealed numerous infiltrates of lymphocytes, histiocytes and plasma cells in the skeletal muscles, sometimes in association with degenerating parasitic cysts and macrophage myophagia. Parasites were not completely eliminated nor prevented from further establishment, therefore the protective immunity was not sterile but rather a state of premunition. PMID- 2967013 TI - [Interaction in the work of an evacuation distribution center and of a provincial health department during World War II]. PMID- 2967014 TI - [Effect of nicotinamide on lipid peroxidation]. AB - Nicotinamide (10-100 mM) caused a decrease in total "fast" flash of chemoluminescence, in rates of NADPH- and ascorbate dependent lipid peroxidation in microsomal fraction of rat liver tissue. Content of cytochrome P-450 (carbonyl complex) as well as rates of amidopyrine N-demethylation and aniline p hydroxylation were also decreased in microsomal fraction. At the same time, inhibition of chemoluminescence was found after addition of 50, 100 mM nicotinamide to blood plasma or to solution of oxidized oleic acid. With an increase in nicotinamide concentration its inhibitory effect on lipid peroxidation was more distinct. PMID- 2967015 TI - [Effect of thymalin on the development of experimental hyperlipidemia and atherosclerosis]. AB - Thymalin, which is a polypeptide preparation from thymus, exhibited hypolipidemic and antiatherosclerotic actions in rabbits maintained on a cholesterol-containing diet during three months. At the same time, thymalin caused favourable effect on the functional activity of lymphocytes impaired in hyperlipidemic animals, i.e. it normalized the T-suppressor activity and sensitivity to atherogenic lipoproteins. PMID- 2967016 TI - [Isolation of poxvirus (Poxviridae, Orthopoxvirus, the cowpox complex) from the root vole Microtus (M.) oeconomus Pal. 1776 in the forest tundra of the Kola Peninsula]. PMID- 2967017 TI - [Successful treatment of a tubal pregnancy by local and systemic administration of prostaglandin]. AB - The recent trend towards conservative management of tubal pregnancies, as well as first reports on the efficacy of systemic or local administration of prostaglandins prompted us to investigate the therapeutic effect of PGF2 alpha injected directly into a non-ruptured tubal pregnancy, followed by systemic administration of a synthetic longer-acting PGE 2 derivative for 3 days. Serial determinations of hormone parameters in the follow up of this patient verified that this combination treatment is capable of dealing with tubal pregnancy without further surgical treatment. Hysterosalpingography performed 6 weeks afterwards showed tubal patency on both sides. PMID- 2967018 TI - [Medical and social status of users of vehicles for the handicapped in a rural district]. PMID- 2967019 TI - Caring for special populations in the dental office. PMID- 2967021 TI - [Percutaneous transluminal angioplasty in abdominal angina--stenosis of the celiac-mesenteric trunk]. PMID- 2967020 TI - Laser Doppler flux and vasomotion in patients before and after transluminal angioplasty for limb salvage. PMID- 2967022 TI - [Examination of rat embryo cell cultures for the occurrence of Mycoplasma pulmonis using an autoradiographic method]. PMID- 2967023 TI - Familial pseudohypoaldosteronism. AB - The clinical course of two siblings with a severe form of pseudohypoaldosteronism was followed over a period of seven and five years respectively. Both children persistently had a high sodium-potassium excretion ratio in the urine, sweat, saliva, and stools as well as high serum concentrations of aldosterone and renin and an increased urinary excretion of tetrahydroaldosterone. Despite sustained treatment with sodium chloride (10-40 mmol/kg/d) and cation exchange resin (sodium polystyrole sulfonate 0.5-2 g/kg/d) they repeatedly developed episodes of salt wasting and hyperkalemia which occurred mainly during uncomplicated respiratory tract infections. Aldosterone receptor characteristics were studied in the cytosol of the rectal mucosa at ages 2.5 years and 6 months respectively. Compared to age matched controls there was a decreased affinity for aldosterone at the low affinity binding site. Among the members of the family, the father and one of his sisters had high concentrations of sodium in the sweat and an increased urinary excretion of tetrahydroaldosterone. PMID- 2967024 TI - Atrial natriuretic peptide and atrial pressures in newborns with transposition of the great arteries. AB - The relation of atrial pressure to atrial plasma concentration of atrial natriuretic peptide (ANP) was studied in four newborns undergoing catheterization because of transposition of the great arteries. In three patients, mean left atrial pressure clearly exceeded right atrial pressure (12 +/- 7 vs. 2 +/- 3 mmHg; mean +/- SD). In one patient mean atrial pressures were identical (4 mmHg). In all patients the ANP concentration in the left atrium exceeded that of the right (833 +/- 464 vs. 415 +/- 366 pg/ml; mean +/- SD). There was a significant (r = 0.97; p less than 0.01) correlation between pressure and ANP concentration in the left atria. In the right atria, no linear correlation existed between pressure and ANP concentration. ANP concentration in the left atrium decreased after a lowering of the left atrial pressure by atrial septostomy. In these patients, the left atrium seems to be the main source of circulating ANP. PMID- 2967025 TI - Concentrations of human atrial natriuretic peptide in the cord blood and the plasma of the newborn. AB - Concentrations of human atrial natriuretic peptide (hANP) in the cord blood and the plasma were measured in 25 newborns. The level of hANP in 0 to 1 post-natal days (212.8 +/- 118.1 pg/ml; mean +/- SD) was significantly higher than that in cord blood (69.7 +/- 53.2 pg/ml) (p less than 0.005). There were no significant differences in the levels of hANP at the ages of 0 to 1, 4 and 6 post-natal days. The level of hANP did not show any significant correlation with urinary excretion of Na, urinary Na/Cr or Na/K ratios. Further evaluations should be made in order to clarify the role of hANP during the early post-natal period. PMID- 2967026 TI - [Simultaneous determination of multi-component vitamins by CPA spectroscopic computing analysis]. PMID- 2967027 TI - [A new polycyclic arylhydrocarbon type inducer of drug metabolizing enzymes--the contragestational agent DL-111-IT]. PMID- 2967028 TI - Atrial natriuretic peptide during exercise in patients with coronary heart disease before and after single dose atenolol and acebutolol. AB - Plasma atrial natriuretic peptide (ANP) was measured during dynamic exercise in 10 patients with coronary heart disease before and after single dose atenolol 50 mg and acebutolol 200 mg, respectively. Systolic blood pressure, heart rate and the rate-pressure product increased during exercise before and after beta blockade, but levels were lower after beta-blockade. Plasma ANP levels at rest were unchanged after atenolol, but rose after acebutolol (p less than 0.01). During exercise plasma ANP increased significantly both before and after beta blockade, but plasma ANP levels were higher after acebutolol at all workloads (p less than 0.05), whereas plasma ANP levels after atenolol were higher at 125 W exclusively (p less than 0.05). The augmented ANP levels during exercise after beta-blockade probably reflect catecholamine-stimulated ANP release, whereas the elevated plasma ANP levels after acebutolol at rest might be a beta-adrenoceptor mediated ANP release due to the intrinsic sympathomimetic effect of acebutolol. PMID- 2967029 TI - Peripheral angioplasty and the newer circulatory interventions: whose responsibility? PMID- 2967030 TI - Maternal age-specific rates of 47,+21 and other cytogenetic abnormalities diagnosed in the first trimester of pregnancy in chorionic villus biopsy specimens: comparison with rates expected from observations at amniocentesis. AB - Results are presented on chromosome analyses made on 4,481 embryos or fetuses studied through chorionic villus sampling (CVS) in whom there was no known bias to presence of a chromosome abnormality except advanced parental age. We excluded from the analysis most cases in which mosaicism was diagnosed or in which there were cytogenetic discrepancies among samples obtained from the conceptus. There remain 48 cases of 47,+21, 39 cases of other nonlethal abnormalities, and 12 lethal abnormalities diagnosed in 4,481 studied. A regression analysis (restricted to the 3,848 cases diagnosed in the 35-49-year maternal age interval) was done on rates of (1) 47,+21, (2) other abnormalities excluding lethals or (3) including them, and (4) all abnormalities excluding lethals or (5) including them. The model used was y = exp(bx + c), where y is the rate of abnormality, x is maternal age at time of CVS (the modal age of the procedure was 10 gestational weeks from the last menstrual period), and b and c were, respectively, (1) 0.288 and -15.527; (2) 0.272 and -15.173; (3) 0.253 and -14.141; (4) 0.282 and -14.753; and (5) 0.271 and -14.195. We also derived rates of abnormalities at the time of CVS that would be predicted from rates (of nonmosaics) at amniocentesis after adjustment for the difference in gestational age between the usual times that these two procedures are done. The difference between the numbers of abnormalities predicted on the basis of these adjusted amniocentesis rates and the numbers observed at CVS provides an estimate of the spontaneous loss of embryos and fetuses between the usual gestational ages of these procedures. In these data, for 47,+21 the estimated proportion lost is 21% but the result is not significant at the .05 level. For other abnormalities excluding lethals the estimated spontaneous loss is 29% (P approximately .05); including lethals it is 44%. For all abnormalities, excluding lethals, pooled together, the estimate is 24%; including lethals it is 33%. The last three values are all significant at the .05 level or lower. The observed rates of abnormalities at CVS would be approximately 10% to 15% higher if one pooled diagnosed mosaics with the nonmosaics, but the estimated proportion of spontaneous fetal loss would be lower. PMID- 2967031 TI - Atrial natriuretic peptide in man. PMID- 2967032 TI - Prognostic value of T lymphocyte subset ratio in idiopathic membranous nephropathy. AB - The behavior of T lymphocyte subsets was studied in 39 Italian patients with nephrotic syndrome due to idiopathic membranous nephropathy. They took part in a long-term prospective and randomized therapeutic trial based on the 6-month administration of methylprednisolone and chlorambucil. The lymphocyte subsets were evaluated by monoclonal antibodies at the beginning of the trial and at the end of the follow-up period in 23 treated and in 16 untreated patients. Our data seem to suggest that a higher helper-inducer/suppressor-cytotoxic cell ratio before therapy may be a good prognostic index of improved proteinuria. Moreover, the therapeutic schedule does not seem to induce a long-lasting abnormality in cellular immunity. PMID- 2967033 TI - Acute renal failure, skin rash, and eosinophilia associated with aztreonam. AB - A case of acute renal failure associated with aztreonam therapy is reported. A man, age 70, with infections due to Klebsiella pneumoniae and Enterobacter cloaca was treated for 9 days with aztreonam and presented with a generalized maculopapular rash, fever, eosinophilia, worsening renal function and increased IgE levels and eosinophiluria. Aztreonam was then discontinued and the patient was treated with supportive measures and steroids and his renal failure reversed over the course of 5 days. This appears to be one of the first cases of acute renal failure associated with aztreonam. PMID- 2967034 TI - Veralipride: alternative antidopaminergic treatment for menopausal symptoms. AB - The vasomotor hot flushes and increased perspiration symptomatic of the menopausal period reflect the adaptation of the body to the lowering of its preset basal temperature resulting from the interference of various central neurotransmitters of suprahypothalamohypophyseal origin. The present double-blind study was conducted to test the efficacy of veralipride, a synthetic antidopaminergic molecule, in eliminating the symptoms of menopause in 50 patients. Results indicated a total elimination of both hot flushes and excessive perspiration in 63% to 80% of the patients treated. The beneficial effects persisted up to 3 months of follow-up. Veralipride significantly increased dehydroepiandrosterone sulfate and estradiol levels. High values of prolactin were found, and some patients showed slight breast discharge; these changes disappeared 48 hours after the drug was stopped. PMID- 2967035 TI - Reappearance of end-diastolic velocity in a pregnancy complicated by severe pregnancy-induced hypertension. AB - A case is reported in which previously absent end-diastolic velocities in the umbilical artery reappeared after treatment in a pregnancy complicated by hypertension. This observation is not consistent with the suggestion that abnormal waveforms are associated with obliteration of the tertiary stem villus arterioles. PMID- 2967036 TI - [Clinico-statistical characteristics of the indicators of physical development of newborn infants in a region with a high birth rate]. PMID- 2967037 TI - [Experience with the organization of perinatal services in the city]. PMID- 2967038 TI - Brain glucose utilization in childhood Huntington's disease studied with positron emission tomography (PET). AB - Brain glucose metabolism was measured in two children with early-onset Huntington's disease, using positron emission tomography with fluorodeoxyglucose (FDG) as the tracer. A marked (48%) hypometabolism was found at the level of the caudate nuclei, but other areas of the brain, particularly the cerebral cortex, were not significantly affected. Despite its different clinical presentation, Huntington's disease in children is characterized by brain metabolic alterations similar to those found in adult patients. PMID- 2967039 TI - The clinical neuromuscular pharmacology of mivacurium chloride (BW B1090U). A short-acting nondepolarizing ester neuromuscular blocking drug. AB - Mivacurium chloride (BW B1090U), a bis-benzylisoquinolinium diester compound, was found to undergo hydrolysis in vitro by purified human plasma cholinesterase in a pH-stat titrator at 88% of the rate of succinylcholine at pH 7.4, 37 degrees C and 5 microM substrate concentration. In 72 consenting ASA Physical Status I-II patients receiving nitrous oxide/oxygen-narcotic-thiopental anesthesia, the neuromuscular blocking effect of mivacurium was assessed following bolus doses from 0.03 to 0.30 mg/kg, as well as during and following continuous infusions from 35 to 324 min in length. The calculated ED95 for inhibition of adductor pollicis twitch evoked at 0.15 Hz was 0.08 mg/kg. At 0.1 mg/kg, 96% block developed, onset to maximum block required 3.8 +/- 0.5 min, and recovery to 95% twitch height occurred 24.5 +/- 1.6 (SE) min after injection. At 0.25 mg/kg, onset was 2.3 +/- 0.3 min; 95% recovery developed within 30.4 +/- 2.2 min, an increase in duration of action of only 24% versus 150% higher dosage. Comparative recovery indices from 5 to 95% or from 25 to 75% twitch heights did not differ significantly among all dosage groups from 0.1 to 0.3 mg/kg (range 12.9 to 14.7 and 6.6 to 7.2 min, respectively). In 38 patients who received mivacurium by continuous infusion (duration 88.1 +/- 7.1/47.1 min, SE/SD) for maintenance of 95 +/- 4% twitch inhibition, the mean 5-95% and 25-75% recovery indices after discontinuation of infusion were 14.4 +/- 0.6 and 6.5 +/- 0.3 min (P greater than 0.5 vs. all single bolus doses). The train-of-four (T4) ratio, within 2.6 +/- 0.5 min after 95% twitch recovery following bolus doses, averaged 79.5 +/- 1.8% (n = 32). Similarly, after discontinuation of infusions, the T4 ratio reached 73.4 +/- 1.9% within 3.4 +/- 1.9 min after 95% twitch recovery (n = 33). Antagonism of residual block was seldom indicated, but, to test ease of reversal, eight patients electively received neostigmine (0.06 mg/kg) with atropine (0.03 mg/kg) at 67 to 93 (76.6 +/- 3.5) % block. Twitch returned to 95% of control within 4.5 to 9.5 (6.3 +/- 0.5) min after neostigmine. Mivacurium may offer increased versatility in providing clinical muscle relaxation in a variety of situations. Further studies seem appropriate. PMID- 2967040 TI - Protein C and S response to danacrine in end-stage renal disease. PMID- 2967041 TI - [Disseminated Bowen's disease 20 years after the handling of radioactive products in the clock manufacture]. PMID- 2967042 TI - [Toxidermatitis after oral pristinamycin]. PMID- 2967043 TI - [Seborrheic eczema (pityriasporosis)]. PMID- 2967044 TI - [What's the significance of the HLA system in dermatology?]. PMID- 2967045 TI - [Intermediate-term trial treatment of arrhythmia in hypertrophic myocardiopathy with sotalol]. AB - Unlike amiodarone, beta-blockers have proved ineffective in the treatment of severe ventricular rhythm disorders encountered in hypertrophic myocardiopathy. We tried to evaluate the efficacy of sotalol, a betablocker with anti-arhythmic properties, like amiodarone, on the supraventricular and ventricular rhythm disorders of this disease. 13 patients (6 men and 7 women, mean age 53.5 +/- 26.5 years) were evaluated with a 48 hours electrocardiographic recording, before and 8 and 120 days after a sotalol treatment at a mean dose of 290 mg/day (160 to 640 mg). The diagnosis of hypertrophic myocardiopathy was established on standard clinical, phonomecanographic (13 cases), sonocardiographic (13 cases) and haemodynamic (9 cases) criteria. Five patients presented bursts of supraventricular tachycardia on DO. Their number decreased to 4 on D8 and to 3 to D120. Six patients presented Lown's class III or IV ventricular rhythm disorders on DO. Their number went from 2 on D8 to 4 on D120. Therefore, in this short series, sotalol has shown a non-negligible efficacy on the rhythm disorders encountered in hypertrophic myocardiopathy. PMID- 2967046 TI - The erythroid membrane skeleton: expression and assembly during erythropoiesis. AB - One aspect of erythropoiesis entails the biogenesis of a membrane skeleton. Analysis of the expression and assembly of the various components of this multisubunit structure illustrates the problem of spatial and temporal regulation of cytoskeletal morphogenesis, departures from simple self-assembly, and the various levels of regulation that might be operative in this aspect of cellular morphogenesis in development. Additionally, it reveals a new insight into the mechanisms underlying the molecular defects in certain hereditary hemolytic anemias. PMID- 2967047 TI - Pathogenesis of membranous nephropathy. AB - Membraneous nephropathy is the most common cause of idiopathic nephrotic syndrome in adults. Recent studies of the pathogenesis of the subepithelial glomerular immune deposits that characterize this disease have revealed new mechanisms of glomerular immune deposit formation involving cell surface antigens and have documented the role of the C5b-9 membrane attack complex of complement in mediating renal injury. Understanding these mechanisms may help us understand the pathogenesis of several other immune-mediated diseases and has implications for possible therapeutic interventions in MN. PMID- 2967048 TI - Targeting a spinal stimulator to treat the 'failed back surgery syndrome'. PMID- 2967049 TI - [Clinical application of monoclonal antibody-drug conjugates in colorectal carcinoma]. AB - Monoclonal antibody, A7, produced from a mouse splenocyte immunized against human colon cancer was used as drug carrier for colon cancer. A7 had not ADCC and ADMC activity but had ACD activity. Anticancer drug, mitomycin C (MMC), and neocarzinostatin (NCS), were covalently bound to A7 to form the conjugates, A7 MMC, and, A7-NCS. In vitro cytotoxic effect of the conjugates on SW1116 was much stronger than that of free MMC or free NCS. The conjugates, A7-NCS, administered in nude mice brought about the highest NCS concentration in tumor, while normal IgG-NCS distributed evenly in all the tissues. The conjugates showed strong antitumor effect on colon cancer transplanted in nude mice. Forty one patients with colorectal cancer including 10 patients with postoperative metastasis were given A7-NCS. The immunoperoxidase and drug concentration studies of the resected specimens revealed that NCS was found to be localized specifically in cancer. There was no serious adverse effect in the patients receiving the conjugate. Of eight patients with postoperative liver metastasis, three showed evidence of tumor reduction on CT scan and three claimed pain relief. The conjugate was of no benefit to the patients with multiple lung metastasis and peritoneal metastasis. PMID- 2967050 TI - [Antitumor activity and toxicity to the immune system and intestine, of the fluorinated pyrimidines FUra, 5'-DFUR, tegafur and UFT]. AB - Antitumor activity and the toxicities of 5-fluorouracil (FUra) and its depot form derivatives with different mechanisms such as 5'-DFUR, tegafur and UFT, were compared in mice, and the therapeutic indices were measured. 5'-DFUR was less toxic to immune organs and the functions than those by other fluorinated pyrimidines. The therapeutic indices of 5'-DFUR obtained by using parameters of particular side effects, reduction of the number of GM-CFU in the bone marrow, shrinkage of the thymus, functional disorders in the immune systems, were several times higher than those of tegafur and UFT. On the other hand, there were no marked differences in the intestinal toxicities among the compounds. 5'-DFUR was only about two times less toxic on the intestinal tract than tegafur and UFT when the incidence-of diarrhea and damage to the duodenum were compared. These studies suggest that the difference in the mechanisms converting to FUra is a result of the toxicities causing different extent. PMID- 2967051 TI - [Comparative studies on the antitumor activity of the fluorinated pyrimidines 5' DFUR, tegafur, UFT and FUra on various murine tumors]. AB - Antitumor activities of fluorinated pyrimidines by oral administration were compared with various murine tumor models. 5'-Deoxy-5-fluorouridine (5'-DFUR) showed a better antitumor activity in terms of the growth inhibition and increase of the survival time than those of 5-fluorouracil(FUra), tegafur and UFT, particularly with respect to the chemotherapeutic indices. The activity of these fluorinated pyrimidines were further investigated in more detail with mice bearing colon 26 adenocarcinoma to suggest some means to optimize their treatment regimens in clinical trials. 5'-DFUR showed the activity irrespective of the size of tumor mass at the time of start of therapy. Additionally only 5'-DFUR was safely administered to the mice daily for long period up to more than 100 days, suppressing the tumor growth and increasing the survival to great extent. PMID- 2967052 TI - [Reductive effect on ascites, of neocarzinostatin in patients with ovarian cancer]. AB - Neocarzinostatin (NCS) was administered intravenously by drop infusion to 16 patients with ovarian cancer, and intrathoracically to one patient. In addition to evaluation of the efficacy of the chemotherapy, we mainly investigated its effect on ascites and pleural effusion, and carried out continuous measurements of the abdominal circumference in 3 patients. NCS relieved thoracic effusion in 1 patient. We observed that abdominal circumference decreased in 10 patients after administration of NCS. The results obtained from continuous measurement of abdominal circumference revealed that NCS significantly reduced ascitic retention in ovarian cancer. PMID- 2967053 TI - [Echocardiography with color Doppler. Another diagnostic method destined for obsolescence?]. PMID- 2967055 TI - [15th National Congress of Cardiology. Villahermosa, Tabasco, 8-12 November 1987. Abstracts]. PMID- 2967056 TI - [Prevention of adverse reactions to drugs]. PMID- 2967054 TI - [Prevention of coronary atherosclerosis. II: Topography and morphology of "protruding" lesions. Epidemiologic, diagnostic and therapeutic implications]. AB - Topographic and morphologic aspects of coronary atherosclerotic "protruding" lesions were investigated in 119 accidental deaths in males living in Mexico City. Morphology and topography varied according to the arterial trunk studied and age. Left anterior descending and circumflex lesions were almost always confined to the initial 5 cms and if a lesion was present after the 3rd cm there was always a proximal lesion. Right coronary lesions occasionally were found distally even in the absence of proximal lesions. Third decade lesions were usually not calcified. Stenosis if present was rarely multivascular. Some fourth decade lesions were calcified; plurivascular stenosis was present in some cases. Fifth decade lesions show sequential stenotic lesions in the same vessel; calcium and plurivascular stenosis were often observed. However the features observed in the 3rd decade could be observed in the older subjects. Reference is made to lesions which are found in both the left main trunk and the left anterior descending. Epidemiological and clinical applications related with preventive programs are mentioned. PMID- 2967057 TI - [Atypical chest pain with normal coronary vessels: clinical study and follow-up of 22 patients]. AB - 22 patients with normal coronarography but with angina pectoris or angina-like chest pain without evidence of coronary spasms are reviewed. We studied clinical characteristics, basal and exercise electrocardiography and other tests used to obtain a diagnosis, although only in one patient were we able to find an esophageal pathology which caused the clinical picture. After 20.7 +/- 14.4 months follow-up no patients had myocardial infarction or acute coronary events. Most of them still experience chest pain, are physically limited and use antianginal drugs. The fact that their coronarograms were normal decreased significantly the number of admissions to hospital (2.4 +/- 3.3 vs 0.26 +/- 0.95, P less than 0.02) and the amount of drugs prescribed. The lack of improvement of most of them seems to justify the need for other studies looking for a positive diagnosis and other types of treatment. PMID- 2967058 TI - [Metabolic disorders in survivors of myocardial infarct]. AB - Several lines of evidence have clearly established the role of lipoproteins as risk factors for the development of atherosclerosis. Epidemiologic studies from different countries have found that about one third of myocardial infarction survivors under 60 years of age are hyperlipidemic. The acute stress reaction occurring in the first hours following an acute myocardial infarction causes distinct changes in the patient's metabolic profile, these changes include a significant reduction of total cholesterol and cholesterol associated with low density lipoproteins and a usually mild elevation of blood glucose. With the purpose of establishing the prevalence and severity of lipoprotein disorders found in myocardial infarction survivors living in Mexico city we conducted a prospective study of 106 consecutive admissions to the coronary care unit at the National Institute of Cardiology with the fully proven diagnosis of acute myocardial infarction, we included only patients younger than 60 years of age that could be sampled within the first 72 hours of the appearance of typical symptoms, at this time the coronary risk factor profile was assessed and blood samples were drawn (acute sample). After three months of the diagnosis we sampled 81 of the original 106 patients (chronic sample). The comparison of these 81 patients showed remarkable differences in the lipid values obtained on each sample. The mean value for total cholesterol in the acute sample was 225 mg/dl whereas the corresponding value for the chronic sample was 240.5 mg/dl (p less than 0.005). This difference was also highly significant for the low density fraction. On the basis of the chronic sample analysis we estimated a prevalence of hyperlipoproteinemia of 35.8%. (II: 18.5%, III: 2.5%, IV: 14.8%), an additional subgroup of 10 patients (12.3%) had the hypo-HDL phenotype raising the number of subjects at risk for atherosclerosis to as high as 48.1% considering only the lipoproteins. The prevalence figures for the rest of the risk factors were as follows: 70.3% for tobacco smoking, 35.8% for Systemic Arterial Hypertension, 33.4% for Obesity and 30.8% for Diabetes Mellitus. Among the group of 81 patients, 17 were known diabetics, eight additional cases of Diabetes Mellitus were diagnosed at the chronic phase (two with fasting hyperglycemia and six with diagnostic oral glucose tolerance tests). The "acute plase" glycemia for these eight subjects was significantly higher (mean: 98.4 mg/dl) than the corresponding value for the non diabetic patients (mean: 83.4 mg/dl p less than 0.002), the seventeen known diabetics had a mean glycemia of 150.6 mg/dl in the acute sample.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967059 TI - [Countercurrent aortography by contrast injection into a peripheral artery]. AB - Countercurrent aortography performed through left brachial artery in 8 children, aging 27 days to 42 months. Systemic-pulmonary anastomosis was possible to be assessed in 3 out of 4 cases, even in one aging 42 months. Aortic coarctation could be evaluated in another 3 patients. In the last case with a clinical suspection of vascular ring, countercurrent aortography disclosed an anomalous right subclavian artery. We concluded that the procedure is safe and useful to assess some conditions involving thoracic aorta, even in older children. PMID- 2967060 TI - [Myocardial infarct without pathologic Q waves]. AB - Neither the presence nor the absence of pathological Q waves are related to the transmural or subendocardial localization of an acute myocardial infarction. The prognosis of an acute myocardial infarction without pathological Q waves is controversial. We studied 23 hospitalized patients with prolonged, typical chest pain, enzymatic elevation, an EKG without pathological Q waves but with changes in ventricular repolarization and a positive scintigraphy with radioactive pyrophosphates. The patients were classified in two groups according to the pattern of pyrophosphate uptake; group A had grade 2+ focal uptake (N = 13) and group B with grade 2+ diffuse uptake (N = 10). The level of serum glutamic transaminase was significantly greater in group A as compared to group B (p less than 0.05). However, levels of creatine kinase and lactic dehydrogenase were no different. The clinical hospital course was similar in both groups. Eleven patients were followed by 31.7 months: 30.4% had chronic angina, one underwent coronary bypass surgery, another suffered a recurrent myocardial infarction and there were no deaths. The diagnosis of acute myocardial infarction should be suspected in patients with prolonged angor pectoris, electrocardiographic changes in ventricular repolarization, increased enzymatic levels and a positive scintigraphy with a grade 2+ diffuse myocardial uptake of Tc 99m pyrophosphates. The diagnosis is certain if the uptake has a focal distribution. PMID- 2967061 TI - [Simultaneous test of ventricular function and myocardial perfusion, during exercise, using a new agent labeled with Tc99m]. AB - One a tribute of a Tc 99m labeled myocardial agent is the possibility to measure both ventricular function and myocardial perfusion with a single injection. To assess this, normal volunteers, 14 patients with coronary artery disease (CAD) and two suffering from cardiomyopathy with normal coronaries, were injected with 8-10 mci carbomethoxy-isopropyl-isonitrile or 20 mci Rp-30 Tc 99m at peak semi recumbent bicycle exercise and again at rest. Thirty msec per frame first pass data, and 5 min static anterior, 40(0-) and 70(0-) left anterior oblique images were obtained. Standard Thallium 201 stress test were also done, within one month, and were at the same level of exercise. The left ventricular ejection fraction (EF) increased with exercise (69%-76%) in normal patients. All studies showed normal myocardial perfusion on exercise. In CAD patients the EF increased in some patients who had ischemia. Perfusion images with Tc 99m during exercise and at rest had an identical correlation with Thallium 201. The results support the concept of dual ventricular function and perfusion studies using a single Tc 99m labelled myocardial agent, and suggest that this could become the standard radionuclide stress tests in the future. PMID- 2967062 TI - [Electrophysiologic characteristics of the transplanted heart]. AB - The electrophysiologic characteristics of the denervated human heart were assessed in 20 cardiac transplants recipient from the Cardiology Department, at the Hospital of San Pablo, Barcelona, Spain. We studied the donor and the recipient sinus node function at rest and exercise test. Holter recording of 24 hours was performed in a few cases. At rest, in 14 patients the intrinsic heart rate of the donor atrium was slower when we used the technique of Jose. When we analyzed all the results we did not find statistical differences in the heart rate of either: the donor or the recipient atria (r = 0.58, p less than 0.01). However we observed a marked increase in heart rate of the donor with exercise test and normal activity (Holter), probably reflecting an increase in circulating catecholamines (exercise test: base-line 116 +/- 16 vs maximum exercise 140 +/- 10, p less than 0.001; Holter: minimal rate 64 +/- 9.66 vs maximal rate 112 +/- 23, p less than 0.001). We also describe the cardiac arrhythmias including two sudden deaths associated with ventricular tachyarrhythmias and in five patients sinus node disfunction in relation with episodes of acute rejection. We concluded that is important to perform a standard 12 lead electrocardiogram, exercise testing, Holter recording and electrophysiologic study as part of the ongoing routine evaluation of surviving cardiac transplant patients. PMID- 2967063 TI - [Hypercapnia induced by acetazolamide and its effect on pulmonary circulation. Experimental study]. AB - In order to evaluate the isolated hemodynamic effects of acute hypercapnea on pulmonary circulation, we designed a canine model in which vascular pressures. PaO2, and arterial pH were maintained within normal limits. Six mongrel dogs were studied. Hypercapnea was achieved by a single intravenous doses of acetazolamide (120 mg/Kg) while maintaining mechanical ventilation constant. Both arterial and mixed venous PCO2 increased from 27 to 41 mm Hg and from 31 to 46 mm Hg respectively, and remained constant through the rest of the experiment (up to 3 Hs). With hypercapnea, total pulmonary vascular resistance increased from 312 +/- 156 to 435 +/- 173 d.s.cm-5 (p less than 0.05) and the stroke index decreased from 20.7 +/- 8.3 to 13.2 +/- 4.8 ml/beat (p less than 0.05). There were no changes either in pulmonary capillary wedge pressure or in the right ventricular end diastolic pressure. The above changes suggest a pulmonary vasoconstrictor effect of hypercapnea in the absence of other known vasoactive factors. PMID- 2967064 TI - [Echocardiographic diagnosis of the right tri-atrial heart]. AB - In this report we describe the two-dimensional echocardiographic features of a case of cor triatriatum dexter. In this situation the right atrium is divided by a membrane in two chambers. Considering the anatomic characteristics of this case we added a new type in the classical classification proposed by Doucette et al. PMID- 2967065 TI - [Infectious Brucella endocarditis in a case of corrected transposition of the great arteries]. AB - We describe a case of a 14 year old male who developed infections endocarditis due to Brucella sp. He had a corrected transposition of great vessels with valvular pulmonary stenosis and mitral insufficiency. Vegetations were demonstrated by echocardiogram on the pulmonary valve as well as on left atrioventricular valve. He had also proliferative nephritis. Prolonged antimicrobial treatment results in complete healing. Because the rarity of this association we inform this observation. PMID- 2967066 TI - [Radiologic characteristics of cardiogenic pulmonary edema in the elderly]. AB - The radiologic appearance of atypical cardiogenic pulmonary edema (ACPE) is presented in 10 cases admitted from 1983 to 1985, with age ranges from 74 to 89, and with diagnosis of ischemic heart disease, with myocardial infarction in 50% of them. Clinically they had asthenia, adynamia and anorexia in 80%, cough and weight loss in 50%. All of them had tachycardia, pulmonary rales and 50% pericardial rub. ECG showed in 80% anterior subepicardial ischemia, 60% posteroinferior subepicardial ischemia, 60% bifascicular block, and 50% left anterior fascicular block. Chest films were interpreted at first as pulmonary fibrosis in 90% of the cases with superior lobe involvement in 50%. Heart enlargement was present in 50%. A chronic lung disease was disclosed on clinical and pulmonary physiological grounds. It is concluded that asthenia, adynamia and anorexia were atypical manifestations of heart failure in the elderly. Silent myocardial infarction was observed in half of our patients and it was complicated with pericardial involvement in 50%. Irregular distribution of fluids in pulmonary edema was attributed to anatomic changes in elder lung. These atypical behaviour of pulmonary edema, has been misinterpreted on radiologic basis with pulmonary infection, tumours, metastasis or fibrosis. Those radiologic changes disappeared or improved in 72 hrs. with treatment of left ventricular failure. PMID- 2967067 TI - [Electrovectocardiographic manifestations of left ventricular and biventricular growth]. AB - The basic criteria for the electrical diagnosis of left ventricular and biventricular enlargements are discussed on the basis of the myocardial depolarization and repolarization sequence. Left ventricular dilatation secondary to isolated diastolic overloading increases the manifestation of the main vectors resulting from the activation of this ventricle. These changes reflect the proximity of the left ventricular walls to the exploring electrodes. The above mentioned vectors appear as tall R waves and wide ventricular curves with counterclockwise rotation on the three planes. If the diastolic overload is a isolated phenomenon, T waves are positive and asymmetric on the left leads while the T loop, of secondary type, is concordant in its orientation with the R loop. This fact is due to a prolonged duration of the repolarization phase of the left ventricle. Global left ventricular hypertrophy produced by a sustained systolic overloading increases the magnitude and manifestation of all the vectors resulting from the depolarization of this ventricle (I, II l, III l) owing to the prolonged duration of the corresponding activation fronts. When LBBB is also present, the first septal vector is not evident. In extreme degrees of the systolic overload, the T wave is inverted and shows morphologic secondary characteristics in left leads, and the T loop opposes the R loop on frontal and horizontal planes. The directional changes of the repolarization fronts of free left ventricular walls can satisfactorily explain these features. Left ventricular hypertrophy of a segmentary type, such as that observed in idiopathic myocardiopathy, generally increases the magnitude and manifestation of septal vector I and II left. When both ventricles are hypertrophied, the electromotive forces originating in the more severely affected heart chamber predominate in electrical records. PMID- 2967068 TI - [Applications of color-coded Doppler 2-dimensional echocardiography]. AB - Conventional Doppler techniques have been used for sometime, but are limited by personal skill and experience, as well as by the fact that it can sample only one specific cardiac site at a time. The introduction of color-coded Doppler flow imaging systems allows by using multiple sample volumes an immediate display of intracardiac blood flow, spatially-oriented and super-imposed in two-dimensional echocardiography images, that not only increase temporal resolution, but which integrates anatomic and functional analysis. Likewise, the objective visualization of abnormal jet lesions permits the application of continuous-wave Doppler echocardiography with a high veracity for derived pressure gradients. Color-coded Doppler echocardiography emerges as the most promising, simple and non-invasive technique for obtaining confident quantitative information in diverse cardiac pathologic conditions and research. PMID- 2967069 TI - [Neuroendocrine changes in chronic cardiac insufficiency]. AB - Throughout the course of chronic congestive heart failure cardiac and peripheral compensatory mechanisms are at play, most of them under the influence of the neuroendocrine system. The reserves of heart rate and contractility are regulated essentially by the noradrenergic system (NAS), but this mechanism is partial and transient owing to the gradual decrease in the density and sensitivity of myocardial beta-adrenergic receptors induced by overstimulation. Adaptation of the heart to exercise may be reduced. This escape phenomenon is also observed with almost all cardiotonic drugs which interfere with cyclic adenosine monophosphate (cAMP), in contrast with the paradoxically favourable effects of beta-blockers in small doses or of drugs that are both agonists and antagonists of beta-adrenergic receptors. The mechanisms which contribute to the induction of left ventricular hypertrophy are imperfectly known. The noradrenergic system and the renin-angiotensin-aldosterone system (RAAS) are probably not the only ones involved. The setting in action of Frank-Sterling heterometric regulation, at first during exercise then permanently, requires an increase in filling pressure obtained by venous constriction (predominantly controlled by the NAS) and, mostly, by an increase in circulating blood volume. NAS and RAAS intervene in the kidneys to produce water-and-salt retention.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967071 TI - [Morphofunctional changes in the myocardium after extensive bilateral resection of the lungs]. AB - In the experiment, performed on 89 dogs, after 67-75% resection of the lungs, functional morphology of the myocardium has been studied by means of histological, electron microscopical and morphometrical methods. Bilateral extensive resections of the lungs are accompanied with hypertrophy of the cardiac muscle, that is revealed at all the levels of its structural organization. Simultaneously, the hypertrophy develops against the background of dystrophic and destructive changes of the myocardium, their depth and extensiveness correlate with duration of the postoperative observations. If an extrapulmonary anastomosis is formed between the inferior lobular pulmonary artery and the corresponding vein before the resection of 67-75% of the lungs, it eliminates the acute overloading of the right ventricle, makes better conditions for development of compensatory-adaptive processes in the myocardium, prevents advancement of the cardio-pulmonary deficiency during the postoperative period. PMID- 2967070 TI - Evaluation of naloxone therapy for Escherichia coli sepsis in the baboon. AB - This study evaluated the effects of naloxone hydrochloride in the treatment of Escherichia coli-induced shock in baboons. The baboons were studied for 12 hours and monitored for survival times. All baboons were intravenously infused for two hours with E coli and treated as follows: group 1, E coli (control); group 2, E coli plus naloxone hydrochloride, 0.5 mg/kg bolus plus 0.5 mg/kg/h for 9.5 hours; and group 3, E coli plus naloxone hydrochloride, 2.0 mg/kg bolus plus 2.0 mg/kg/h for 3.8 hours. Naloxone was administered after arterial pressure had reached the nadir (more than two hours following initiation of E coli infusion). Mean arterial pressure was supported by the lower dose of naloxone; however, sustained leukopenia and neutropenia were not reversed by its infusion. Naloxone prevented the increase in plasma beta-endorphin level and blunted the increase in plasma cortisol level. Despite these effects, naloxone did not prevent multiple-organ disease and did not decrease mortality. PMID- 2967072 TI - [Coronary transluminal angioplasty: evaluation by the stress test associated with myocardial scintigraphy with thallium-201]. PMID- 2967073 TI - Lp(a) phenotyping by immunoblotting with polyclonal and monoclonal antibodies. AB - A new method that allows rapid phenotyping of genetic Lp(a) glycoprotein types in large numbers of samples is described. The method is based on sodium dodecyl sulfate gel electrophoresis of reduced serum or plasma in horizontal slab gels followed by immunoblotting with polyclonal anti-Lp(a) lipoprotein or monoclonal anti-Lp(a) glycoprotein antibodies. Phenotyping of 194 unrelated, healthy subjects resulted in Lp(a) allele frequencies of Lp(a)B = 0.013, Lp(a)S1 = 0.032, Lp(a)S2 = 0.106, Lp(a)S3 = 0.096, Lp(a)S4 = 0.156, and Lp(a)O = 0.600, and confirmed the recently recognized association of Lp(a) glycoprotein phenotype with Lp(a) lipoprotein concentration. The new procedure is suitable for large scale population, genetic, and epidemiologic studies and may be important for atherosclerotic risk assessment. PMID- 2967074 TI - The International Classification of Impairments, Disabilities and Handicaps: its usefulness in classifying and understanding biopsychosocial phenomena. AB - The International Classification of Impairments, Disabilities and Handicaps (ICIDH) was developed because of the growing awareness that ICD-9 failed to reflect many of the problems that people bring to health care systems. The ICIDH was designed to classify the consequences of disease such as the disruption of daily activity and the social disadvantage that accompany illness. However, since its introduction, the ICIDH has largely been ignored by psychiatry, despite its conceptual strength. In particular, it provides a framework for applying the biopsychosocial model and studying the phenomena of mental illness. The latter is an important issue because of clinical psychiatry's inherent weakness in distinguishing between symptoms of disease and the psychosocial consequences, a particular inadequacy of the DSM-III diagnostic criteria. The utility of the ICIDH is shown in the study of the phenomena of panic disorder and agoraphobia, where I conclude that agoraphobia should be classified as a disability/handicap and not a disorder. PMID- 2967075 TI - Absence of back pain and tachycardia in the emergent presentation of abdominal aortic aneurysm. PMID- 2967077 TI - [Psychiatric morbidity in primary care centers]. PMID- 2967078 TI - Down's syndrome in Hyderabad, India. AB - A cytogenetic analysis of Down's syndrome revealed straight trisomy 21 in 96.5 per cent affected. Remaining 3.5 per cent were found to have chromosomal mosaicism. In addition, a normal karyotype was observed in 2 individuals with Down's phenotype. A survey of Down's syndrome in Hyderabad gave an incidence of 1.17 per 1000 or 1 in 853 live births. There was a significant increase in the mean maternal age of the affected over the controls. Sex ratio was also higher in our sample of Down's syndrome as compared to the general population. PMID- 2967076 TI - Inhibition of thymidylate synthase in intact L1210 cells by ara-C, daunomycin, hydroxyurea and 3,4-dihydroxybenzylamine. AB - The use of an in situ assay for thymidylate synthase has shown that a variety of clinically important drugs, including arabinofuranosylcytosine, hydroxyurea, and daunomycin, inhibit thymidylate synthase in intact cells. In contrast to the inhibition observed with 5-fluorodeoxyuridine, inhibition occurs by an indirect mechanism, is delayed in onset, and is incomplete. Inhibition occurred at concentrations that corresponded to those that inhibit DNA synthesis, suggesting that this phenomenon might contribute to the biological action of these agents. Since the inhibition of thymidylate synthase by this indirect mechanism appears to be a general property of drugs that inhibit DNA synthesis, our findings may have important implications for the mechanism of killing of tumor cells, as well as the rationale for combination regimens. PMID- 2967079 TI - Atrial natriuretic factor in pregnancy-induced hypertension and preeclampsia: increased plasma concentrations possibly explaining these hypovolemic states with paradoxical hyporeninism. AB - Plasma immunoreactive atrial natriuretic factor 99-126 (ir ANF), plasma volume, plasma renin activity, and plasma aldosterone were measured during pregnancy in 14 normotensive nonpregnant women, 15 normotensive pregnant women, 35 patients with pregnancy-induced hypertension (PIH), and in ten patients with preeclampsia (PE). Repeated measurements were carried out 2 months after delivery in a subgroup of the same patients. The plasma levels of ANF were found to be higher in pregnant normotensive women than in nonpregnant normotensive women, but the decrease of plasma ANF 2 months after delivery was not significant on the basis of seven paired data, so that it cannot presently be stated with certainty that pregnancy per se stimulates ANF secretion. Still higher levels of ANF were found in PIH and, especially, in PE. A positive correlation was found in the pooled population of normotensive and hypertensive pregnant women between plasma ANF and mean arterial pressure. A greater decrease of plasma ANF was found after delivery in the hypertensive patients than in the normotensive controls. This excludes an absolute deficiency of ANF secretion in the pathogenesis of hypertension. These findings suggest a compensatory role of ANF in the prevention of blood pressure increase. Plasma renin activity (PRA) and plasma aldosterone concentrations were higher in normotensive pregnant women than in normotensive nonpregnant women. Compared to normal pregnancy, plasma volume was decreased in PIH (-17%) and in PE (-25%), whereas PRA was less increased in both groups and plasma aldosterone concentration was less increased only in the PE group. The simultaneous high levels of plasma ANF may explain this inappropriate hypostimulation of renin secretion by hypovolemia in these hypertensive states. PMID- 2967080 TI - Atrial antinatriuretic factor in the developing Dahl hypertensive rat. AB - In order to determine the developmental pattern of atrial concentrations of atrial natriuretic factor (ANF) in the Dahl hypertension-prone rat, atrial ANF concentrations were measured in inbred hypertension-prone (S/JR) and hypertension resistant (R/JR) Dahl rats at 5, 15, 25, and 51 days of age. In both strains, atrial ANF concentrations peaked at 15 days of age. Atrial ANF concentrations did not differ between the two strains from 5 to 25 days of age. However, by 51 days of age, atrial ANF concentrations in the S/JR rat were significantly greater than those of the R/JR rat. Combining these data with developmental patterns of plasma renin activity in S/JR rats suggests the possibility that the S/JR rat may become intravascularly volume-expanded between 25 and 51 days of age. This volume expansion may contribute to the etiology of hypertension in this model of essential hypertension. PMID- 2967081 TI - Expression of heat shock genes of Neurospora crassa: effect of hyperthermia and other stresses on mRNA levels. AB - Neurospora crassa mycelium was heat shocked for intervals varying from 15-180 min. Heat shock mRNA was monitored by hybridization of Northern blots with the Drosophila hsp-70 gene probe and an inducible member of the yeast hsp-70 gene family, YG100. A 2.7 kilobase (kb) transcript, with homology to these two probes, was detected in cultures shocked for 15 min; its levels increased up to 60-90 min and declined thereafter. Sodium arsenite, too, induced the synthesis of this transcript. An additional, constitutively synthesized 2.4-kb transcript was revealed by hybridization with the yeast probe. The synthesis of this message was terminated during heat shock. Hybridization of Northern blots with the Drosophila actin gene probe demonstrated two size classes, 1.85 and 1.63 kb; the former decreased dramatically following heat shock. Recovery, as assessed by the disappearance of the 2.7-kb hsp-70-mRNA and restoration of the 1.85-kb actin message to the prestress levels, was essentially complete within 60 min of transfer to 28 degrees C. In vitro translations of RNA from stressed cells showed the heat shock messages to be stable and readily translatable. RNA of cells subjected to heat shock plus CdCl2 showed a higher content of messages for heat shock proteins of 70, 80, and 90 kilodaltons. PMID- 2967082 TI - Directional probability information and Down syndrome: a training study. AB - In previous studies we have found that subjects with Down syndrome did not respond to changes in directional probability within a complex motor task in the manner of other subjects with mental retardation (Blais & Kerr, 1986; Kerr & Blais, 1985). Only after extended practice were significant changes noted in their reaction time (RT) performance; however, movement time (MT) data remained relatively unchanged (Kerr & Blais, 1987). In the present study, subjects were given training relative to the directional probability component of the task (choice of direction). The subjects with Down syndrome did improve their subsequent performance, but primarily in terms of MT. Although they did adopt a pattern of responding to changes in directional probability (RT) similar to that of subjects without mental retardation, the nature of the change reflected the extent of previous exposure to the task. PMID- 2967083 TI - The use of preventive health services by disabled children. AB - As a measure of the use of general pediatric health services we assessed disabled children's receipt of preventive health care in relation to a control group of nondisabled children, matched by age, family size, and region of residence. The study and control subjects were identified in a household survey conducted in Minnesota in 1976 and ranged in age from 1 to 18 years. The proportion who made a preventive health visit was nearly identical in both groups. Binary variable multiple regression showed that 9 percent of the variation in outcome was explained by the independent variables, which included demographic and socioeconomic characteristics and access to health care. Disability did not contribute significantly to the explained variation, but family structure, mother's education, and mother's use of preventive health services reached the 95 percent level of significance. The results suggest that children in a community who are identified as disabled are not at a disadvantage, in comparison with the nondisabled, in gaining access to preventive health services. The use of such services by all children appears to be low when information on using school health services is not available. PMID- 2967084 TI - The HIV positive patient. PMID- 2967085 TI - Assessment of left ventricular performance during percutaneous transluminal coronary angioplasty: a study by intravenous digital subtraction ventriculography. AB - Left ventricular performance during percutaneous transluminal coronary angioplasty was assessed in 52 patients by intravenous digital subtraction ventriculography. After injection of contrast into the right atrium ventriculograms were obtained before and during balloon inflation. In 37 patients they were also obtained after the procedure. A 12 lead electrocardiogram was monitored throughout. During balloon inflation the left ventricular ejection fraction fell (from 73% to 57%) in all but one patient; the decreases in patients with single vessel or multivessel disease were similar. The fall in left ventricular ejection fraction during percutaneous transluminal coronary angioplasty of the left anterior descending artery (19%) was significantly greater than that during balloon inflation in the right coronary (10%) or circumflex (8%) coronary arteries. It also reduced anterobasal, anterior, and apical segmental shortening while right coronary percutaneous transluminal coronary angioplasty affected inferior and apical segments. In 33 (63%) patients the ST segment was altered during balloon inflation. The fall in left ventricular ejection fraction correlated significantly with the magnitude of both ST segment elevation (r = 0.637) and ST depression (r = 0.396). Left ventricular ejection fraction and regional wall motion returned to baseline values after the procedure. Balloon inflation during percutaneous transluminal coronary angioplasty produces considerable abnormalities of global and regional left ventricular performance and this indicates the presence of myocardial ischaemia, which may not be apparent on electrocardiographic monitoring. Intravenous digital subtraction ventriculography is useful for monitoring left ventricular performance during controlled episodes of coronary occlusion produced by balloon inflation. PMID- 2967087 TI - Patterns of diastolic dysfunction in left ventricular hypertrophy. AB - The relative sensitivities of and interrelations between different measurements of diastolic function were studied in 50 patients with left ventricular hypertrophy diagnosed on anatomical grounds. Isovolumic relaxation time, the interval from minimum cavity dimension to mitral valve opening and relative dimension increase during this period, and the peak rate of dimension increase and wall thinning during rapid ventricular filling were measured by digitised M mode echocardiography. The relative heights of peak early diastolic and atrial velocities (a/E) and the time for decline of early diastolic velocity to half its peak value (velocity half time) were measured on continuous wave and pulsed Doppler and the relative height of the "a" wave was measured by apexcardiogram. All sets of values except those of the interval from minimum dimension to mitral opening were unimodally distributed, and all differed significantly from those in 20 age matched controls. The relative height of the "a" wave on the apexcardiogram (90% values were abnormal) was the most sensitive method of studying left ventricular diastolic function and peak rate of dimension increase was the least sensitive. Though none of the correlations was high, there were individual associations between peak rate of dimension increase, a/E, peak wall thinning rate, and velocity half time, and independently between delay in mitral valve opening and dimension change during this period. Other values seemed to be independent of one another, suggesting a different physiological basis. It is concluded that these various abnormal values do not reflect a single underlying disturbance of diastolic function. There are at least four possible discrete abnormalities: prolongation of isovolumic relaxation; incoordination during isovolumic relaxation; reduced rate of rapid filling; and an increase in the relative amplitude of the "a" wave probably caused by increased passive stiffness. These may be present singly or in combination in any patient. PMID- 2967086 TI - Perforation thresholds and safety factors in in vivo coronary laser angioplasty. AB - Laser angioplasty can cause early (acute perforation) or late (stenosis or aneurysm) complications. To find how much intravascular laser energy can be delivered via a 100 microns core optical fibre passed down a balloon angioplasty catheter without causing angiographic abnormalities up to 10 days later, argon laser energy was delivered percutaneously under radiographic screening to the coronary circulation of 12 normal closed chest dogs. With the balloon inflated, sequential laser pulses were delivered to the same site. Angiograms were recorded before, immediately, and again at one week, after laser delivery. There were two laser-induced perforations (both fatal). Mechanical perforation with the 100 microns fibre occurred four times, but there were no haemodynamic sequelae. To find the acute perforation threshold of similar sized arteries to energy delivered via the bare 100 microns core fibre, the tip of which was held in contact with the luminal surface, 32 argon laser pulses were delivered transluminally in vivo to separate sites in normal rabbit iliac and canine coronary arteries. The acute perforation threshold with energy delivered via the angioplasty catheter lay between 6 and 10 J and that without the balloon angioplasty catheter lay between 3 and 4 J. After delivery of up to 6 J via a balloon angioplasty catheter, there were no angiographic abnormalities at one week. Fibre optic transluminal delivery of laser energy may improve the primary success rate of, and perhaps widen the indications for, coronary angioplasty. PMID- 2967088 TI - Plasma atrial natriuretic polypeptide concentrations during and after reversion of paroxysmal supraventricular tachycardias. AB - Plasma concentrations of immunoreactive atrial natriuretic polypeptide were raised in 22 of 23 patients with paroxysmal supraventricular tachycardia and in all seven patients with atrial flutter. Plasma concentrations of atrial natriuretic polypeptide rose soon after the onset of supraventricular tachycardia. A sample taken 30 minutes after reversion to sinus rhythm (pharmacological or non-pharmacological) showed a significant fall in 19 of the 23 patients with paroxysmal supraventricular tachycardia and all seven patients with atrial flutter. Because atrial natriuretic polypeptide has powerful natriuretic and diuretic properties, an increase may contribute considerably to the polyuria that is often associated with episodes of supraventricular tachycardia. PMID- 2967089 TI - Disposition of fosinopril sodium in healthy subjects. AB - 1 Fosinopril sodium is the first phosphorus-containing angiotensin-converting enzyme (ACE) inhibitor to be studied clinically as an antihypertensive agent. It is an ester prodrug that is hydrolysed in vivo to the active diacid ACE inhibitor, SQ 27, 519. 2 In a three-way crossover study, nine healthy male subjects (age range 20-34 years) each received an intravenous 7.5 mg dose of SQ 27, 519-[14C] and two oral 10 mg doses of [14C]-fosinopril sodium, administered as a capsule and in solution. 3 After the intravenous dose of SQ 27, 519, the 0 to 96 h recovery of radioactivity averaged 44 and 46% of the dose in urine and faeces, respectively, indicating substantial biliary secretion. Only intact SQ 27, 519 was detected in the plasma, urine, and faeces following the intravenous dose of SQ 27, 519. 4 After oral doses of fosinopril sodium, about 75% of the radioactivity in plasma and urine was present as SQ 27, 519; the remainder corresponded mainly to a beta-glucuronide conjugate of SQ 27, 519 (15-20%), and a monohydroxylated analogue of SQ 27, 519 (about 5%). Negligible amounts of fosinopril sodium were present, indicating complete hydrolysis of the prodrug. 5 For the solution and capsule doses, respectively, the oral absorption of fosinopril sodium averaged 32% and 36% and the oral bioavailability of SQ 27, 519 averaged 25% and 29%. 6 The average values for clearance (39 ml min-1), renal clearance (17 ml min-1), Vss (10 1), and plasma protein binding (approximately 95%), indicated that SQ 27, 519 was slowly cleared from the body and not distributed extensively into extravascular sites. PMID- 2967090 TI - Complement factors in fetal and maternal blood and amniotic fluid during the second trimester of normal pregnancy. AB - Complement factors (C3, C4, C5; Factors B, H and I) were measured in maternal and fetal serum and amniotic fluid obtained from 55 women with singleton pregnancy undergoing diagnostic fetoscopy at 15 to 28 weeks gestation. Maternal serum levels were consistently 10 times higher than fetal levels which in turn were 10 times higher than levels in amniotic fluid. Spearman rank correlation analysis at weeks 20 to 22 (n = 20) revealed a statistically significant correlation between maternal and fetal levels of C3 and Factors B and I, and between maternal and amniotic fluid levels of Factors B and I. A significant increase in fetal levels of C3, C4 and Factor H, and in amniotic fluid levels of C3 and Factor B was seen in relation to advancing gestational age. These differences were not seen in maternal serum during the short interval of pregnancy studied. These data confirm earlier assumptions of fetal synthesis of complement factors, and provide normal reference ranges of complement factors in fetal blood and amniotic fluid. PMID- 2967091 TI - Cell volume-sensitive Na+-ATPase activity in rat kidney cortex cell membranes. AB - A ouabain-insensitive, K+-independent, sodium pump, has been demonstrated in guinea-pig and rat kidney proximal tubular cells. This pump is thought to be distinct from the ouabain-sensitive Na+/K+ pump. We present evidence here indicating the modulation of the biochemical expression of the Na+ pump, i.e. the ouabain-insensitive Na+-ATPase, by the cell volume in rat kidney proximal tubular cells. Thus, basolateral plasma membranes from swollen cells show a ouabain insensitive Na+-ATPase activity 10-times higher than that in membranes from control cells. If the swollen cells recover their volume, the activity decreases ten times to control values. The ouabain-sensitive Na+/K+-ATPase is not affected by changes in the cell volume. PMID- 2967092 TI - A role for Ca2+ in mediating hormone-induced biphasic pepsinogen secretion from the chief cell determined by luminescent and fluorescent probes and X-ray microprobe. AB - In isolated chief cells from the guinea pig, cholecystokinin (10 nM) and a high concentration of ionomycin each caused a biphasic pattern of pepsinogen secretion. The initial fast response to cholecystokinin was not dependent on medium Ca2+ ans was mimicked by low concentration of ionomycin (100 nM). Inositol 1,4,5-trisphosphate caused a similar fast release from permeabilized cells. The slow component of release was dependent on medium Ca2+, however, and was mimicked by the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) (100 nM) or the diacylglycerol analogue 1-oleoyl-2-acetylglycerol (OAG) (100 microM). Ionomycin (100 nM) and TPA (and/or OAG), when applied together, reproduced the biphasic pattern of pepsinogen secretion, suggesting that the signalling pathways utilized by both types of agonist contribute to the response evoked by cholecystokinin hormone stimulation. Both fura-2 and aequorin were used to monitor changes of intracellular Ca2+. Three pathways were found to contribute to the Ca2+ transient. A rapid release of Ca2+ from intracellular store(s), a rapid Ca2+ entry from the extracellular space, and a more sustained Ca2+ entry from the extracellular space. Cholecystokinin induced a rapid increase in cytoplasmic Ca2+ ([Ca2+]i) as estimated with fura-2 and aequorin. This rise was reduced but not abolished upon removal of extracellular Ca2+, suggesting that both Ca2+ entry from the extracellular space and Ca2+ mobilization from the intracellular store(s) contribute to the initial, fast component of the Ca2+ transient. A second, more sustained component of the Ca2+ transient induced by cholecystokinin was abolished by lanthanum. TPA and OAG induced a biphasic Ca2+ transient that could be detected only with aequorin. The late, sustained component of this response was again abolished by lanthanum as well as by removal of extracellular Ca2+. It appears that the late component of the Ca2+ transient is dependent on Ca2+ influx from the extracellular space and is too localized to be detected by fura-2. Prestimulation of cells with TPA or OAG prevented the aequorin transient caused by cholecystokinin and vice versa, suggesting that TPA, OAG and cholecystokinin activate the same pathways of Ca2+ entry into the cytosol from the intracellular store(s) or the extracellular space. The stimulation-sensitive Ca2+ pool was examined with electron probe X-ray microanalysis. It appears to be restricted to an area enriched in secretory granules or peripheral endoplasmic reticulum just beneath the apical plasma membrane and in close association with the microtubular-microfilamentous system.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967093 TI - Serial [18F]N-methylspiroperidol PET studies to measure changes in antipsychotic drug D-2 receptor occupancy in schizophrenic patients. AB - An indirect approach to the relationship among drug dose, plasma level, and the competition between a labeled neuroleptic drug [18F]N-methylspiroperidol (18F NMS) for binding sites in striatal tissue in normal and schizophrenic subjects is described. The slope of the line plotting the ratio of activity in the striatum (As) to activity in the cerebellum (Ac) versus time up to 5 hr postinjection of 18F-NMS is taken as a marker of site occupancy. An inverse relation between labeled competitor uptake and drug plasma level has been demonstrated for the classes of antipsychotic drug studied. Striatal uptake studies showed a progressive increase in all subjects following drug withdrawal up to 156 hr postwithdrawal. Uptake and clearance of 18F-NMS in cerebellar tissue was not appreciably affected by antipsychotic medication or drug withdrawal. PMID- 2967094 TI - Inhibition of the inductive activity of demineralized bone matrix by different percutaneous implants. AB - Teflon tubes with either Gore-Tex (PTFE) or Dacron felt sleeves were implanted percutaneously in rats, in conjunction with demineralized bone matrix. This resulted in different inflammatory reactions, as well as in chondrogenesis and osteogenesis in the subcutaneous tissues. Although induction of osteogenesis by the demineralized bone matrix in the vicinity of the foreign material was inhibited, circumferential bone formation was highly reproducible. The prospect of utilizing demineralized bone matrix in order to enhance acceptance of percutaneous tubes is discussed. PMID- 2967095 TI - Ambiguous phenotypes and genotypes in 16 children with acute leukemia as characterized by multiparameter analysis. AB - Ambiguous phenotypes and genotypes were observed in 16 children with acute leukemia. Surface marker, cytogenetic, molecular genetic, and DNA flow cytometric analyses as well as standard morphologic and cytochemical studies were used to divide the patients into three groups. The first group comprised five children with acute leukemia whose blast cells were morphologically lymphoid, while immunophenotyping disclosed simultaneous expression of early pre-B cell and myeloid features. Molecular genetic studies showed evidence of heavy-chain immunoglobulin (Ig) gene rearrangements in all patients. Cytogenetic data, available in three of these children, revealed t(4;11). In five of the 16 patients, morphologic and surface marker analyses indicated the coexistence of two separate cell populations, one with myeloid and the other with early pre-B cell features. Further evidence of B cell commitment in these patients was provided by demonstration of Ig heavy-chain gene rearrangements in all five patients. Surprisingly, one of the five patients showed oligoclonal Ig heavy chain as well as monoclonal gene rearrangement for the beta chain of the T cell receptor (beta-TCR). The last group consisted of four cases with otherwise typical acute lymphoblastic leukemia (ALL), early pre-B cell phenotype, and coexpression of myeloid or T cell-associated antigens, and two children with unequivocal acute myeloid leukemia (AML) and coexpression of T cell antigens. Gene rearrangement of Ig heavy-chain could be demonstrated in five of six patients, additional Ig light-chain gene rearrangement in two children with ALL, and bigenotypic features (Ig heavy-chain and beta-TCR gene rearrangement) in one patient. In none of the 16 patients did flow cytometry disclose clonal abnormalities of leukemic cell DNA content. Based on these findings, we suggest that malignant transformation in the first and second group of patients took place at a stage ontogenetically close to the pluripotent stem cell, whereas ambiguous phenotypes in the third group resulted from aberrant gene expression or insufficient reagent specificity. PMID- 2967096 TI - Defective helper function of purified T4 cells and excessive suppressor activity of purified T8 cells in patients with B-cell chronic lymphocytic leukemia. T4 suppressor effector cells are present in certain patients. AB - We investigated helper and suppressor functions to B-cell responses and T-T cell interactions of purified T4 and T8 cells from 20 untreated patients with B-cell chronic lymphocytic leukemia (CLL). Appropriate mixtures of purified T4 or T8 cells from patients with CLL were cultured with purified B cells or T4 and B cells from normal donors for 7 days with pokeweed mitogen (PWM). IgM, IgA, and IgG produced were determined in the supernatants of these cultures by a heavy chain-specific enzyme-linked immunosorbent assay (ELISA) and compared to those obtained by the corresponding mixtures of T4, T8, and B cells from normal donors. Purified T4 cells from 14 of 20 patients with CLL exhibited defective helper function (P less than .001) to immunoglobulin (Ig) production by purified B cells from normal donors. Purified T4 cells from 6 of these 14 patients were able to suppress significantly (P less than .001) and in a concentration-dependent manner Ig production by mixtures of T4 and B cells from normal donors, in the absence of T8 cells. These suppressor effector T4 cells from certain patients were partially radiosensitive. Purified T8 cells from 8 of 20 patients with CLL exhibited excessive suppressor activity. These cells significantly suppressed (P less than .001), Ig production by mixtures of T4 and B cells from normal donors to a degree significantly higher (P less than .005) than that observed by equal numbers of T8 cells from normal donors. This inhibition was dependent on the numbers of the T8 CLL cells added to the cultures. Excessive suppressor activity by T8 CLL cells was at least in part radiosensitive in four of eight patients. These results demonstrate a wide range of immunoregulatory T-cell abnormalities in patients with CLL. Naturally occurring T4 suppressor effector cells, directly inhibiting Ig production by mixtures of T4 and B cells, in the absence of T8 cells, are present in certain patients with CLL. PMID- 2967097 TI - A study of the action of amlodipine on adrenergically regulated sodium handling by the kidney in normotensive and hypertensive rats. AB - 1. An investigation was undertaken to examine the effect of calcium channel blockade, induced by amlodipine, on the ability of the renal sympathetic nerves to cause an antidiuresis and anti-natriuresis in normotensive Sprague Dawley and spontaneously hypertensive rats anaesthetized with pentobarbitone. 2. Low frequency renal nerve stimulation in normotensive rats, which did not change renal blood flow, caused a 15% reduction in glomerular filtration rate and was associated with falls in urine flow of 37%, absolute sodium excretion of 47%, and fractional sodium excretion of 38%. The magnitude of these renal excretory changes was unaffected by prior administration of amlodipine at either 200 micrograms kg-1 plus 50 micrograms kg-1 h-1 or 400 micrograms kg-1 plus 100 micrograms kg-1 h-1. Amlodipine given in the higher dose, decreased basal levels of blood pressure and increased basal urine flow and sodium excretion. 3. In spontaneously hypertensive rats, renal nerve stimulation minimally affected renal haemodynamics but decreased urine flow, absolute and fractional sodium excretion by 29%, 31% and 24%, respectively. 4. Similar renal nerve stimulation in spontaneously hypertensive rats given amlodipine at 200 micrograms kg-1 plus 50 micrograms kg-1 h-1 or 400 micrograms kg-1 plus 100 micrograms kg-1 h-1 caused minimal changes in renal haemodynamics and in the excretion of water and sodium. The higher dose of drug resulted in decreased blood pressure and increased basal rates of urine flow and sodium excretion. 5. These data show that in spontaneously hypertensive rats but not normotensive rats, calcium channel blockade inhibited the ability of the renal nerves to stimulate the reabsorptive processes for sodium at the renal tubule. This indicated that in spontaneous hypertension the post-receptor mechanisms had changed and become more dependent on the inward movement of calcium. PMID- 2967098 TI - Morphine does not stimulate prolactin release during lactation. AB - The ability of morphine to stimulate prolactin and growth hormone (GH) release was investigated in male rats and in female rats during diestrus, proestrus and lactation. In agreement with previous reports, acute morphine administration produced an increase in circulating levels of prolactin in male and in diestrous and proestrous female rats. In contrast to these results, morphine administration (10 or 15 mg/kg, s.c.; 5 mg/kg, i.v.; 5 or 10 micrograms, i.c.v.) did not produce an increase in prolactin levels in lactating dams. Morphine stimulates prolactin release in part by decreasing dopamine turnover in the tuberoinfundibular neurons in the median eminence. In order to assess the functional activity of these neurons during lactation, haloperidol (0.1 or 0.5 mg/kg, i.v.) was given to lactating dams. There was a significant increase in prolactin levels following haloperidol administration, suggesting that these dopaminergic neurons are participating in the modulation of prolactin release during lactation. In contrast to the insensitivity of the lactating rat to morphine stimulation of prolactin release, the intraventricular administration of two other opiate receptor agonists, beta-endorphin (10 or 20 micrograms) and [D-Ala-D Leu]enkephalin (DADLE; 5 or 10 micrograms), produced significant increases in circulating levels of this hormone. The GH response to morphine, beta-endorphin and DADLE was also measured in these same rats. All these opiate receptor agonists stimulated GH release in male rats and in female rats during diestrus and proestrus as well as during lactation. These observations suggest that the suckling stimulus during lactation renders the rat refractory to morphine stimulation of prolactin release, possibly as a result of down-regulation of the mu-opiate receptor subtype. PMID- 2967099 TI - The insulin-like growth factor-II (IGF-II) receptor of rat brain: regional distribution visualized by autoradiography. AB - The presence of insulin-like growth factor-II (IGF-II) in brain and cerebral spinal fluid prompted us to investigate the distribution of receptors for this peptide in rat brain slices. Human 125I-IGF-II (10 pM) was incubated for 16 h at 4 degrees C with thaw-mounted slices of rat brain from 11 different brain regions. Incubations in the absence or presence of excess unlabeled human IGF-II or insulin were performed and the labeled tissues were exposed to X-ray film for 4-7 days. Autoradiographs showed dense labeling in the granule layers of the olfactory bulbs, deep layers of the cerebral cortex, pineal gland, anterior pituitary, hippocampus (CA1-CA4, and dentate gyrus), and the granule cell layers of the cerebellum. Unlabeled IGF-II eliminated most of the binding in these brain regions while insulin produced only a minimal reduction in the amount of 125I-IGF II bound. These results indicate that a neural receptor for IGF-II is uniquely distributed in rat brain tissue supporting the notion that this peptide might play an important role in neuronal functioning. PMID- 2967100 TI - Intraparenchymal fetal striatal transplants and recovery in kainic acid lesioned rats. AB - Striatal kainic acid (KA) lesions induce behavioral and biochemical deficits which resemble symptoms encountered in patients suffering from Huntington's disease. In rats with KA lesions, fetal striatal transplants have shown to reverse the pervasive nocturnal hyperactivity induced by the lesion. In the present study 4.6 mm3 of fetal striatal tissue were delivered bilaterally into the anterodorsal portion of the lesioned caudate nucleus. Care was taken to deliver the transplant within the host parenchyma and away from the lateral ventricles. Locomotor behavior analyzed using the Digiscan animal activity monitors before and after the transplants demonstrated a reversal of the hyperactivity following transplants in 70% of lesioned animals. Microinjections of horseradish peroxidase delivered into the globus pallidus and substantia nigra of a small group of functionally recovered transplanted animals, did not reveal evidence for reinnervation between host nigra or pallidum and the transplant at 10 weeks post-transplantation. Other laboratories have reported anatomical connections by 6 months post-transplantation. Ventricular/brain ratios demonstrated that intraparenchymal transplants significantly reduced the ventricular dilation following KA lesion. These results suggest that functional recovery can be obtained when the transplant is immersed into the host's striatal parenchyma regardless of the existence of long-range anatomical connections. PMID- 2967101 TI - Three cases of diflunisal hypersensitivity. PMID- 2967102 TI - Are physicians' opinions about chiropractors changing? PMID- 2967103 TI - Biological activities of 5-fluorouracil and its prodrug 5'-deoxy-5-fluorouridine in rats. AB - The antitumor activity and toxicity of 5-fluorouracil (FUra) and 5'-deoxy-5 fluorouridine (dFUR) were compared in female Fischer rats bearing transplanted dimethylhydrazine-induced colon tumors. The therapeutic effects of FUra and dFUR were not affected by the initial tumor size, but were dependent on the dose and duration of treatment. The maximal response rate of 80-90% cures was obtained with 7-day infusions of 35 mg-kg-1-day-1 FUra or 500 mg-kg-1-day-1 dFUR. The host toxicity of FUra and dFUR in tumor-bearing or normal rats included gastrointestinal and central nervous system disturbances. Toxicity related death was preceded by a greater than 20% animal weight loss and other signs of gastrointestinal disturbances. The maximal therapeutic dose of FUra was identical to the toxic dose which caused 40% death in normal rats. By contrast, the maximal therapeutic dose of dFUR did not cause toxic death, and the threshold lethal dose of dFUR was 40% higher than the maximally therapeutic dose, indicating a better therapeutic index for dFUR in this rat tumor. PMID- 2967104 TI - Removing the barriers. PMID- 2967105 TI - Laminin-dependent and laminin-independent adhesion of human melanoma cells to sulfatides. AB - Sulfatides (galactosylceramide-I3-sulfate) but not neutral glycolipids or gangliosides adsorbed on plastic promote adhesion of the human melanoma cell line G361. Direct adhesion of G361 cells requires densities of sulfatide greater than 1 pmol/mm2. In the presence of laminin, however, specific adhesion of G361 cells to sulfatide or seminolipid (galactosylalkylacyl-glycerol-I3-sulfate) but not to other lipids is strongly stimulated and requires only 25 fmol/mm2 of adsorbed lipid. The effects of laminin and sulfatide on adhesion are synergistic, suggesting that laminin is mediating adhesion by cross-linking receptors on the melanoma cell surface to sulfatide adsorbed on the plastic. Although thrombospondin binds to sulfatides and G361 cells, it does not enhance, but rather inhibits direct and laminin-dependent G361 cell adhesion to sulfatide. In contrast, C32 melanoma cells also adhere specifically to sulfatide, but adhesion of these cells is not enhanced by laminin or inhibited by antibodies to laminin that block laminin-dependent adhesion of G361 cells. Thrombospondin is a potent inhibitor of C32 cell adhesion to sulfatide. Fucoidan, which inhibits laminin binding to sulfatide, inhibits laminin-dependent adhesion of G361 cells by 50% at 0.2 micrograms/ml. Several other tumor cell lines also attach directly on sulfatide-coated surfaces. Laminin stimulates adhesion to sulfatide of three of the six cell lines tested. The ability of laminin to promote adhesion of tumor cells to sulfatide suggests that binding to sulfatide could participate in laminin-mediated cell-cell adhesion. Thus, many tumor cell lines can attach on sulfatide substrates using endogenous sulfatide binding proteins, and in some cells laminin but not thrombospondin can promote tumor cell adhesion to sulfatide. PMID- 2967106 TI - A newly designed angioplasty catheter: "the Gemini balloon". AB - A triple-lumen angioplasty composite balloon polyethylene catheter that incorporates a central lumen to ultimately accommodate a 0.035-inch wire is described. The two additional lumens are responsible for either simultaneous or independent expansion of the angioplasty balloons. Balloon dimensions vary from 3 mm through 8 mm, with any of the combinations being possible. Its principal indication will be in those extremely tight lesions that initially require dilatation with the lower-profile balloon followed by the more appropriate, larger-sized, balloon, thus eliminating the necessity of catheter exchange and an additional balloon. PMID- 2967107 TI - Suppressor activation in autoimmune thyroiditis in mice. PMID- 2967108 TI - Down syndrome and Cu/Zn-SOD: an additional interpretation. PMID- 2967109 TI - Mitochondrial protein import: identification of processing peptidase and of PEP, a processing enhancing protein. AB - Transport of nuclear-encoded precursor proteins into mitochondria includes proteolytic cleavage of amino-terminal targeting sequences in the mitochondrial matrix. We have isolated the processing activity from Neurospora crassa. The final preparation (enriched ca. 10,000-fold over cell extracts) consists of two proteins, the matrix processing peptidase (MPP, 57 kd) and a processing enhancing protein (PEP, 52 kd). The two components were isolated as monomers. PEP is about 15-fold more abundant in mitochondria than MPP. It is partly associated with the inner membrane, while MPP is soluble in the matrix. MPP alone has a low processing activity whereas PEP alone has no apparent activity. Upon recombining both, full processing activity is restored. Our data indicate that MPP contains the catalytic site and that PEP has an enhancing function. The mitochondrial processing enzyme appears to represent a new type of "signal peptidase," different from the bacterial leader peptidase and the signal peptidase of the endoplasmic reticulum. PMID- 2967110 TI - Purification and characterization of two forms of rat interleukin-2. AB - Rat IL-2 produced by spleen cells in culture with concanavalin A was purified using gel filtration, hydrophobic chromatography, and ion-exchange chromatography. At least two forms of rat IL-2 were found to be separable by ion exchange chromatography. These two forms have been designated form I and form II. Form I of rat IL-2 was purified by a factor of 1297 and found to have a pI of 6.4. Form II was purified by a factor of 669 and found to have a pI between 5.4 and 6.1. Lectin chromatography was used to demonstrate that these two forms most likely differ in the extent of glycosylation. In the presence of tunicamycin the production of form II was significantly reduced. The two forms of rat IL-2 differ in their abilities to promote a mixed-lymphocyte reaction. Their differences in glycosylation may be the reason for these differences in activity. PMID- 2967111 TI - Similarities between LAK cells derived from human thymocytes and peripheral blood lymphocytes: expression of the NKH-1 and CD3 antigens. AB - Human thymocytes are devoid of NK cells but develop lymphokine-activated killer (LAK) activity after culture with recombinant interleukin-2 (rIL-2). The most active precursor for this activity appears to be a CD3-negative cell. The purpose of these studies was to compare the phenotype and functional activities of thymocyte and peripheral blood lymphocyte (PBL) LAK cells. Following culture, rIL 2-activated thymocytes resemble PBL-generated LAk and PBL NK cells. For each of these populations, lytic activity is highest in NKH-1-positive cells. Two-color fluorescence of each population also indicates that NKH-1+ cells are highly granular, as measured by staining with the lysosomotropic vital dye quinacrine. PBL, PBL-derived LAK cells, and thymus-derived LAK cells have a portion of cells that express both CD3 and NKH-1. However, approximately 60-80% of NKH-1+ cells lack detectable CD3. This suggests that both CD3+ and CD3- cells may be capable of LAK activity. Thymic-derived LAK cells respond to interferon in a manner very similar to NK and PBL-derived LAK cells, but lack the NK-associated CD16 antigen. Thus, despite the absence of NK cells in the thymus, it is possible to generate thymocyte LAK activity which bears a strong resemblance to LAK activity derived from peripheral blood lymphocytes. PMID- 2967112 TI - [Etiopathogenic importance of anaerobic non-sporulating bacteria in maxillary sinusitis]. PMID- 2967113 TI - [Detection of hepatitis B surface antigens using monoclonal antibodies]. PMID- 2967114 TI - [Determination of immunocomplexes. III. A method using a solid surface for immunocomplex binding and use of C1q subcomponent of the first complement component (C1q SP)]. PMID- 2967115 TI - [Practical aids for the diagnosis of urogenital chlamydiosis]. PMID- 2967116 TI - [Initial experience with the detection of Campylobacter pylori]. PMID- 2967117 TI - [Use of the latex agglutination reaction in the identification of coagulase positive and coagulase-negative Staphylococci]. PMID- 2967118 TI - [An epidemic of enterotoxicosis from ice cream containing staphylococcal TSST-1]. PMID- 2967120 TI - [Moellerella wisconsensis--the first isolation and identification of a new genus and species of the family Enterobacteriaceae in Czechoslovakia]. PMID- 2967119 TI - [The hemolytic properties of strains of Morganella morganii]. PMID- 2967121 TI - [Microbicidal effects of Chlordetal--a new chlorine disinfective agent recommended for a single-stage method of disinfection]. PMID- 2967122 TI - Regulation of the trehalose-6-phosphate synthase complex in Saccharomyces. I. Interconversion of forms by phosphorylation. AB - Trehalose-6-phosphate synthase is another example of an enzyme of carbohydrate metabolism, in Saccharomyces, which could be regulated by interconversion of forms. Deactivation was mediated both in vivo and in vitro by a cyclic AMP dependent protein kinase. Reversibility of this process was obtained by a phosphatase treatment leading to an increase in activity. The phosphorylated, less active form of the enzyme proved to be more susceptible to activation by ATP.Mg. Mutants with well defined lesions in the cyclic AMP-dependent protein kinase system were used to corroborate our findings of a possible regulatory mechanism of trehalose-6-phosphate synthase activity by interconversion of forms. PMID- 2967123 TI - Regulation of synthesis and secretion of acid and alkaline phosphatases in Neurospora crassa. AB - We show that N. crassa represses the production of acid phosphatase at pH higher than 8.0, irrespective of the carbon source used, whereas production was stimulated by sucrose at slightly acidic pH. The same profile of acid phosphatase production was observed in the pho-2A, pho-3A, nuc-1A, nuc-2A and pregc mutant strains. We also show that acid phosphatase synthesized by the pregc mutant strain grown on high phosphate medium has pronounced differences when compared to the enzyme synthesized by the wild-type strain grown on low phosphate medium in terms of heat stability, steady-state kinetic properties and DEAE-cellulose chromatography. In addition, the synthesis and/or secretion of only phosphate repressible alkaline phosphatase is affected by mutations in acu-1, and acu-5 and acu-7 genes. These results, which indicate distinct pathways for the synthesis and secretion of acid and alkaline phosphatases in N. crassa, contradict the dosage titration model proposed by Metzenberg et al. (1974) whereby the synthesis of these enzymes should occur through a single hierarchical regulatory circuit as a response to phosphate starvation. PMID- 2967125 TI - Enzyme histochemical and morphological phenotype of amphophilic foci and amphophilic/tigroid cell adenomas in rat liver after combined treatment with dehydroepiandrosterone and N-nitrosomorpholine. AB - Male Sprague-Dawley rats were investigated after N-nitrosomorpholine (NNM) treatment with concomitant and subsequent administration of dehydroepiandrosterone (DHEA) for development of pre-neoplastic and neoplastic liver lesions. In addition to clear, acidophilic, mixed cell and basophilic foci, a hitherto undescribed lesion type demonstrating a unique morphological and histochemical phenotype was observed in animals receiving both NNM and DHEA. The cells of the majority of these lesions for which we propose the designation amphophilic foci were characterized by increased granular acidophilia and randomly scattered cytoplasmic basophilia. Histochemically, reduced glycogen content and elevated activity of glucose-6-phosphate dehydrogenase (G6PDH), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), acid phosphatase (AP), succinate dehydrogenase (SDH) and catalase (CAT) were evident. The lack of gamma glutamyl transpeptidase (GGT) or glutathione S-transferase placental form (GST-P) in foci of this type allowed clear differentiation from other NNM-induced focal lesions while suggesting certain similarities to pre-neoplastic cells induced by hypolipidemic agents. Similar enzyme histochemical patterns were characteristic for foci and later appearing nodules (adenomas) composed of amphophilic/tigroid cells the basophilic material of which was increased and frequently arranged in long striped bands. DHEA treatment, while not itself inducing any preneoplastic foci, was thus associated with altered phenotypic expression of foci and adenomas generated by NNM. PMID- 2967124 TI - Plasmid recovery from transformants and the isolation of chromosomal DNA segments improving plasmid replication in Neurospora crassa. AB - The efficient recovery of plasmid DNA from Neurospora crassa transformants is described. Lithium acetate-treated spores were transformed with plasmid DNA and grown in mass in liquid culture. The resulting mycelial growth was harvested and plasmid DNA was extracted and used to transform E. coli to ampicillin resistance. Although at low frequency, routine recovery of plasmid pSD3 which carries the Neurospora qa-2+ gene and pBR322 sequences has been demonstrated. About 10% of the recovered plasmids carried deletions and transformed Neurospora at a higher frequency. The liquid culture procedure was also used in attempts to isolate autonomously replicating sequences (ars). In order to select for a stable vector which contains an ars sequence, a clone bank containing a selectable marker (qa 2+) and Neurospora chromosomal BamHI fragments was constructed and used to transform Neurospora. Several plasmids isolates resulting from a screening of the clone bank showed an improvement in the efficiency of recovery from Neurospora transformants. The properties of one such isolated plasmid, pJP102, suggest that it may contain an ars sequence. Some potential applications of these results for cloning in Neurospora and other filamentous fungi are discussed. PMID- 2967126 TI - Dehydroepiandrosterone and two structural analogs inhibit 12-O tetradecanoylphorbol-13-acetate stimulation of prostaglandin E2 content in mouse skin. AB - Dehydroepiandrosterone, a naturally occurring adrenal steroid, is a highly effective tumor chemopreventive agent in laboratory mice and rats, inhibiting spontaneous breast cancer and chemically induced tumors of the lung, colon, skin, liver and thyroid. Dehydroepiandrosterone blocks three processes that have been implicated in experimental tumorigenesis: (i) carcinogen activation through the mixed-function oxidases, (ii) 12-O-tetradecanoylphorbol-13-acetate stimulation of superoxide anion production in neutrophils, and (iii) 12-O-tetradecanoylphorbol 13-acetate stimulation of [3H]thymidine incorporation in mouse epidermis. All of these effects of dehydroepiandrosterone very likely result from glucose-6 phosphate dehydrogenase inhibition and a lowering of the NADPH cellular pool. It is now reported that oral administration of dehydroepiandrosterone (0.2% in the diet) for two weeks inhibits the stimulation in prostaglandin E2 content in mouse epidermis produced by topical application of 12-O-tetradecanoylphorbol-13 acetate. Two synthetic steroids, 16 alpha-fluoro-5-androsten-17-one and 16 alpha fluoro-5 alpha-androstan-17-one, which are more potent inhibitors of the above three processes in tumorigenesis and are also more effective than dehydroepiandrosterone in inhibiting skin papilloma development in the mouse, are more active in suppressing prostaglandin E2 induction by 12-O-tetradecanoyl phorbol-13-acetate. These two structural analogs, which also lack specific side effects associated with dehydroepiandrosterone treatment, may find application as cancer chemopreventive drugs in humans. PMID- 2967127 TI - Serum Lp(a) level as a predictor of vein graft stenosis after coronary artery bypass surgery in patients. AB - Although the serum lipoprotein fraction Lp(a) has been associated with coronary artery atherosclerosis, its relationship to narrowing of saphenous vein grafts has not previously been elucidated. We therefore measured serum Lp(a) levels in 167 symptomatic patients undergoing cardiac catheterization who had had coronary artery bypass surgery 0.7 to 14.3 years earlier. Lp(a), total cholesterol, and total triglyceride levels were compared with the degree of saphenous vein graft stenosis to test for any association. Serum Lp(a) levels were significantly associated with the degree of stenosis of saphenous vein grafts (r = .24, p = .002). Mean Lp(a) levels (mg/dl) in the 135 patients with stenosis were almost double (32.0 +/- 32.7, mean +/- SD) those in the 32 patients with no graft stenosis (16.7 +/- 22.6; p = .002). Graft stenosis was not associated with previous myocardial infarction, hypertension, obesity, diabetes, or smoking. Serum cholesterol levels (mg/dl) were slightly higher in the stenosis group (251.3 +/- 69) than in the no-stenosis group (231.8 +/- 48.8), but the difference was of borderline significance (p = .06). A stepwise increase in mean Lp(a) was found in groups of patients with increasing vein graft stenosis. At a serum Lp(a) level of 31.6 mg/dl or above, 92% of the patients demonstrated vein graft stenosis. Thus, patients with elevated Lp(a) levels have an increased risk of developing saphenous vein graft stenosis after coronary bypass surgery. PMID- 2967128 TI - Time course of regression of left ventricular hypertrophy after aortic valve replacement. AB - To assess the time course and extent of regression of myocardial hypertrophy after removal of the inciting hemodynamic stress, 21 patients with either aortic stenosis or aortic insufficiency were studied preoperatively, after an intermediate period (1.6 +/- 0.5 years), and late (8.1 +/- 2.9 years) after aortic valve replacement, and results were compared with those in 11 control patients. After aortic valve replacement there was significant hemodynamic improvement, with a fall in the left ventricular end-diastolic volume index (164 +/- 73 to 105 +/- 35 ml/m2, p less than .01), a fall in left heart filling pressure (19 +/- 9 to 12 +/- 5 mm Hg, p less than .01), and maintenance of the cardiac index (3.3 +/- 0.8 to 3.5 +/- 0.8 liters/min/m2, NS) and left ventricular ejection fraction (60 +/- 13% to 64 +/- 10%, NS). By the late study the cardiac index (4.0 +/- 0.6 liters/min/m2, p less than .01) and left ventricular ejection fraction (66 +/- 15%, p less than .05) had further increased and were significantly greater than before surgery. For the group as a whole, the left ventricular muscle mass index fell 31% after surgery by the time of the intermediate postoperative study (174 +/- 38 vs 120 +/- 29 g/m2, p less than .01), and a further 13% from the intermediate to the late study (105 +/- 32 g/m2, p less than .05). At the preoperative study left ventricular muscle mass index was greatest in those patients with aortic insufficiency (191 +/- 36 g/m2), and greater in those with aortic stenosis (158 +/- 33 g/m2) than in control subjects (85 +/- 9 g/m2, p less than .05). At the intermediate postoperative study left ventricular muscle mass index remained significantly higher in both those with preoperative aortic insufficiency (128 +/- 29 g/m2) and those with stenosis (114 +/- 27 g/m2) than in the control subjects (p less than .01). By the time of the late postoperative study there were no longer any significant differences in left ventricular muscle mass index. Thus, the regression of myocardial hypertrophy is a process that occurs over many years after correction of the primary hemodynamic abnormality. As this process of myocardial remodeling occurs, continued improvement in cardiac function may occur, and the improvement occurring between the intermediate and late postoperative studies at a slight but constant afterload excess (inherent in the relative stenosis of the aortic prosthesis) suggests that the hypertrophied myocardium is operating at a reduced level compared with normal myocardium. PMID- 2967129 TI - Interaction of prostaglandins and angiotensin II in the modulation of renal function in congestive heart failure. AB - Despite a dramatic fall in renal blood flow, glomerular filtration rate is usually preserved in patients with congestive heart failure until the terminal stages of the disease. This maintenance of renal function appears to be achieved in part by the synthesis of two vasoactive factors within the kidney--angiotensin II and prostaglandins--which are rapidly released whenever renal perfusion is compromised or sympathetic nerve traffic to the kidneys is increased. Although these two hormonal systems exert opposite effects on systemic and renal blood flow and sodium and water excretion, both act to preserve glomerular filtration rate: prostaglandins by a vasodilator action exerted primarily on the afferent arteriole and angiotensin II by a vasoconstrictor effect on the efferent arteriole. Consequently, when the synthesis of these hormones is experimentally blocked, renal function deteriorates, especially in subjects with marked renal hypoperfusion and sodium depletion; these two factors interact to determine the importance of intrarenal hormonal release in the modulation of renal function. Clinically, four specific factors have been identified that predispose patients with heart failure to the development of functional renal insufficiency after treatment with converting-enzyme or cyclo-oxygenase inhibitors: (1) marked renal hypoperfusion, (2) vigorous diuretic therapy, (3) diabetes mellitus, and (4) intensity of hormonal inhibition within the kidney. This last risk factor may provide the basis for differentiating among enzyme-inhibitory drugs and suggests that renal insufficiency in low-output states may be minimized by the development of therapeutic agents that block hormonal synthesis selectively at sites that are critical to the disease process but spare the homeostatic tissue-based enzyme systems that exist within the kidney. PMID- 2967130 TI - Use of polyethylene glycol in radioimmunoassays involving tritium labels. PMID- 2967131 TI - Rapid, simplified radioimmunoassay of arginine-vasopressin and atrial natriuretic peptide in plasma. AB - We have improved a radioimmunoassay for arginine-vasopressin (AVP) and atrial natriuretic peptide (ANP) by using Sep-Pak C18 cartridges to extract AVP and ANP from acidified plasma. The analytes are co-eluted by use of a mobile phase consisting of 1,2-dimethoxyethane and 40 g/L aqueous trifluoroacetic acid (95/5, by vol). After rapid evaporation of the solvents, AVP and ANP are assayed by a nonequilibrium radioimmunoassay method in which commercially available antibodies and radiolabeled antigens are used. The bound fractions are separated from the free by use of polyethylene glycol with human gamma globulin and rabbit anti human IgG as the second antibody. This results in very low nonspecific binding: 0.44% for the ANP assay, 0.70% for AVP. The minimum detectable amount of ANP is 0.39 pg per tube; for AVP, it is 0.13 pg per tube. Compared with other published methods, this method is substantially more reliable, economical, and easily established in a clinical chemistry laboratory. PMID- 2967132 TI - Secretion of atrial natriuretic peptide and digoxin-like immunoreactive substance during pregnancy. AB - We investigated the secretion of atrial natriuretic peptide (ANP) and digoxin like immunoreactive substance (DLIS) during pregnancy, labor, and the puerperium, as measured in maternal and umbilical cord plasma. There were no significant changes in maternal concentrations of ANP during all three periods, and the concentrations were similar to those found in nonpregnant controls and in umbilical cord plasma. Maternal concentrations of DLIS increased significantly in the second half of pregnancy, peaked during labor, then decreased abruptly within 24 h of expulsion of the infant and placenta to values approaching the nonpregnant range. DLIS concentrations in umbilical cord plasma, however, were significantly higher than in maternal plasma during labor. The abrupt fall in DLIS in maternal plasma and the absence of a significant difference in DLIS concentrations between arterial and venous cord plasma suggest that, during pregnancy, the fetus, not the placenta, is the source of DLIS in maternal plasma. PMID- 2967133 TI - Effects of haemolysis and prolonged cold storage of human plasma on the alpha atrial natriuretic peptide concentration. PMID- 2967134 TI - An enzymatic method for the alpha-amylase assay which comprises a new procedure for eliminating glucose and maltose in biological fluids. AB - An alpha-amylase assay in biological fluids, characterized by a new procedure for eliminating glucose and/or maltose, was developed. The reagent includes thermostable glucokinase, glucosephosphate isomerase and phosphofructokinase obtained from a thermophile Bacillus stearothermophilus. Up to 4,000 mg/dl glucose or 600 mg/dl maltose had no effect on the measured value of alpha-amylase activity when measured at 37 degrees C, even at a serum volume fraction in the reagent of 1/50. Alpha-Amylase activity was monitored by the absorbance increase at 340 nm due to NADPH production. The assay has a high degree of precision, with the within-run and day-to-day coefficients of variation being 2.17% at 75.0 U/l and 2.49% at 112 U/l, respectively, and is linear up to about 2,500 U/l. Regarding interferences, bilirubin, urate, ascorbate, pyruvate, EDTA, sodium fluoride and others were found to have no effect on the assay, and the reagent in solution is stable for about 2 wk at low temperatures (6-8 degrees C). PMID- 2967135 TI - Alpha-form of atrial natriuretic peptide released from dog atrium during atrial distension. AB - To assess the released form of atrial natriuretic peptide (ANP) during atrial distension, we compared the form of ANP before and during atrial distension in anesthetized dogs, using radioimmunoassay combined with high performance liquid chromatography (HPLC). When increasing the left atrial pressure (LAP) from 2.2 +/ 0.9 mmHg to 9.4 +/- 1.3 mmHg, the plasma ANP concentration in the coronary sinus was increased by 190% over the control, during left atrial distension. The major form of the released ANP extracted in the blood sample obtained from the coronary sinus was alpha-form. However other unidentified immunoreactive peaks, preceding and following the main peak were also discernible. These results suggest that the major form of ANP released during left atrial distension was the alpha-form. PMID- 2967136 TI - Inhibition of antisuppression by the acute murine graft-versus-host reaction. AB - Profound suppression of both humoral and cell-mediated immunity is a significant systemic effect of graft-versus-host reactions. Although no complete explanation has been advanced for this immunosuppression suppressor cells have been implicated. The data presented in this paper indicate that acute GVH reactions in (C57BL/6J X A/J) F1-hybrid mice induced by the injection of A/J cells severely disrupts the function of the antisuppressor T-cell pathway at both its induction and effector stages. Results show that within 3 weeks of induction of the reaction, Ly1+-T antisuppressor inducer cells lose their ability to generate the serum factor that mediates antisuppression. This factor is normally taken up by and activates Ly2+ T cells which then inhibit suppressor T-cell function. The data also reveal that Ly2+ T cells collected 2 weeks after induction lose their ability to be activated by the antisuppressor factor produced in normal mice. These cells are thus unable to function as antisuppressor effector cells. The uptake of the antisuppressor factor by Ly2+ T cells depends on the expression of Ia antigens on the surface of these cells. Experiments have shown that these antigens are absent from the surface of T cells derived from mice with GVH reactions. This finding may provide an explanation for the inability of these cells to function as antisuppressor effectors. Antisuppression is an important T cell pathway that is intimately associated with the regulation of immune function. It is possible that the immunosuppression arising in mice with GVH reactions may stem, in part, from unopposed suppressor T-cell activity that results from widespread interference by the reaction with a pathway that normally inhibits suppressor cell activity. PMID- 2967137 TI - Sex differences in regulation of contact sensitivity reaction in mice. 1. Influence of sex on the generation of contrasuppressor and afferent suppressor cells. AB - It is well known that humoral and cell-mediated immune responses are better in females than in males. Females also develop autoimmunity more easily than males. Contact sensitivity, one of the forms of cell-mediated immunity, is controlled at the afferent and efferent phases by complex interactions of regulatory T cells. Our present experiments indicate that T suppressor afferent (Ts-aff) and T contrasuppressor cells (Tcs) are generated in the mouse in a sex-dependent fashion. These two types of regulatory cells are induced by antigen-antibody complexes containing various immunoglobulin isotypes. Females require fewer antigen (Ag)-IgG1 complexes to produce Tcs cells, but more Tcs cells after antigenic stimulation in females tips the balance toward better immune responsiveness. It remains to be established whether the peculiarities in generation of regulatory cells in female mice are relevant to the pathogenesis of autoimmune diseases which predominantly affect females. PMID- 2967138 TI - Impairments in functional subsets of T-suppressor (CD8) lymphocytes, monocytes, and natural killer cells among asbestos-exposed workers. AB - Peripheral blood leukocytes from asbestos-exposed workers were analyzed by dual color flow cytometry using monoclonal antibodies that identify developmental (HLA DR) and functional (Leu 8) subsets of T helper, suppressor lymphocytes, and monocytes. An increase in the number of T suppressor cells was closely associated with a decrease in T lymphocyte functions while numerical defects in activated monocytes (Leu M3+Ia+) and natural killer cells (Leu 7+) were correlated with a depressed Th/Ts ratio. Furthermore, among asbestos-exposed workers with depressed T cell functions we have demonstrated a significantly higher number of the effector Ts (Leu 2+ Leu 8-) subset which regulates both the Th/Ts lymphocyte system as well as B cells and NK cell activities. These findings identified changes in the T suppressor feedback regulatory loop as being responsible for the immunoregulatory imbalance among long-term asbestos workers. In double blind analyses of demographic and radiographic data these phenotypic changes were not correlated with age, smoking history, or duration of exposure but were associated with radiographic evidence of asbestos-associated effects. This correlation established a direct link between asbestos exposure and the subsequent development of immune dysfunction. PMID- 2967139 TI - A renorenal suppression in response to atrial natriuretic peptide in the unilateral ischaemic rat. AB - 1. The present study was carried out to investigate the haemodynamic and natriuretic effects of atrial natriuretic peptide (ANP) in normal rats and in rats with unilateral ischaemia. 2. Twenty-four hour unilateral ureter occlusion gave rise to a marked vasoconstriction in the ipsilateral kidney after its release. Intravenous infusion of ANP doubled p-aminohippurate clearance and inulin clearance and elicited massive natriuresis in the hydronephrotic kidney, while in the contralateral kidney these clearance values were decreased and there was a lack of natriuresis. The responses in the latter kidney were also different from those in the normal rat, in which significant natriuresis was elicited by ANP even though a decrease in p-aminohippurate clearance occurred. 3. After the control kidney in the unilateral hydronephrotic rat was denervated either mechanically or by pretreatment with prazosin, ANP induced a natriuresis in both kidneys. Furthermore, the renal denervation prevented the decrease in inulin clearance in the control kidney after ANP administration. 4. These findings suggest that the renal response to ANP may depend on the vascular tone before administration, and that renal nerve activity may modify the effects of ANP on renal haemodynamics and sodium excretion. PMID- 2967140 TI - Psychological factors and metabolic control: time for reappraisal? PMID- 2967141 TI - Application of modelling techniques to the assessment of insulin sensitivity in man. PMID- 2967142 TI - Early-onset type 2 diabetes vs maturity-onset diabetes of youth: evidence for the existence of two discrete diabetic syndromes. PMID- 2967143 TI - Type 2 diabetes in Arab patients in Kuwait. AB - We describe the characteristics of 75 Arab Type 2 diabetic patients in Kuwait. Their age (+/- SD) at onset was 41 +/- 10 years, and fasting serum C-peptide concentration was 0.32 +/- 0.23 nmol/l (n = 51). Fifty-three percent (37/70) possessed HLA-DR3 or -DR4 epitopes, and 64% (47/73) had a family history of diabetes. Data review suggested that they could be segregated into two groups, those under 40 years old at onset (32 +/- 6 years, n = 37), and the remainder (48 +/- 6 years, n = 38) (p less than 0.001). Those in the former group had a significantly higher frequency of a family history of diabetes than those in the latter group (92% vs 38%, p less than 0.001) suggestive of a greater genetic influence on the development of Type 2 diabetes in those with early onset disease. PMID- 2967144 TI - A measure of treatment satisfaction designed specifically for people with insulin dependent diabetes. AB - The psychometric properties of a diabetes-specific treatment satisfaction scale were examined with responses from 128 adults with insulin-dependent diabetes who had used one of three treatment options for a period of 12 months. The reliability of the seven-item measured was found to be satisfactory (Cronbach's Alpha = 0.76) and factor analyses indicated three useful sub-scales (Perceived General Management; Perceived Compatibility with Lifestyle; Perceived Frequency of Hypo/hyperglycaemia). Use of the treatment satisfaction measure in a feasibility study of continuous subcutaneous insulin infusion (CSII) demonstrated the measure's ability to distinguish between three treatment groups (CSII, intensified conventional therapy and conventional therapy). People choosing to use CSII reported significantly greater improvements in satisfaction than those choosing either from of conventional therapy (F = 36.6; df 2, 125; p less than 0.001). If used in conjunction with measures of blood glucose control, the Treatment Satisfaction measure offers the opportunity for a more holistic appraisal of outcomes in studies evaluating and comparing treatments for insulin dependent diabetes. PMID- 2967145 TI - Insulin compatibility with polymer materials used in external pump infusion systems. AB - In a study designed to mimic actual user conditions for external insulin pump infusion, the insulin quality after passage through the infusion set was assessed by various analytical methods, including high performance liquid chromatography. The two infusion sets tested consisted of, firstly, a polyvinylchloride/rubber syringe and a polyvinylchloride catheter sterilized by gamma irradiation and, secondly, a polyethylene/polypropylene syringe connected to a polyethylene catheter and sterilized by ethylene oxide. The insulin solution delivered through the PVC infusion set showed a reduction of preservative to less than 30% of the initial content and increased formation of chemical transformation products of insulin varying from twice the reference level during the first day to more than three times on the third day. By contrast, the polyethylene/polypropylene infusion system showed only a minor decrease in preservative content and no increase in chemical transformation. These effects were observed irrespective of the brand of insulin and were not affected by increase of the zinc content of the insulin solution. Investigation of the influence of the sterilization methods performed on polyvinylchloride and polyethylene catheters revealed that gamma irradiated polyvinylchloride catheters were markedly harmful to the insulin solution, whereas ethylene oxide sterilization did not influence the chemical stability of insulin. PMID- 2967146 TI - An amperometric needle-type glucose sensor tested in rats and man. AB - An amperometric glucose-measuring 25 gauge (0.5 mm diameter) needle-type sensor has been developed using a glucose oxidase and dimethyl ferrocene paste behind a semi-permeable membrane situated over a window in the needle. Electron transfer results in direct current generation. Sensors have been tested subcutaneously in the abdomen both in anaesthetized rats (40 sensors, 11 rats) and in normal, conscious man (20 sensors, 10 subjects). In rats the blood glucose was modulated by glucose and by insulin infusion. In man the glucose concentrations were rapidly changed by use of a glucose clamp at 12 mmol/l plasma concentration for 2 h, after which the glucose returned to normal. In rats the median correlation between glucose change was 0.83 with an interquartile range from 0.70 to 0.92, and in man the median correlation was 0.80 with an interquartile range 0.67 to 0.86. Hysteresis, a measure of the accuracy on the upswing and downswing, was not a problem and cross-correlation showed no phase-lag. There were quantitative differences between in vitro calibration and the performance in vivo, reflecting the different conditions of use. The current in response to a glucose concentration was stable over 6.0 h in rats and 4.5 h in man. PMID- 2967147 TI - The prevalence of proteinuria detected by Albustix in a defined diabetic population. AB - All the known diabetic patients (917) from a defined population (90,660) were called for review by a single observer. A total of 842 (92%) attended and proteinuria, identified using Albustix (0.3 g/l or more), was found in 57 cases (6.8%), but in 9 this was in association with a urinary tract infection. Diastolic blood pressure, an ulcerated or amputated lower limb, and smoking category were found to be the only significant predictors of proteinuria after a multiple logistic regression analysis. A serum creatinine greater than 150 mumol/l was found in 29 (3.8%) of the 768 diabetics in whom it was measured. However, proteinuria was only present in 7 of the diabetics with impaired renal function. In those aged less than 65 years, the prevalence of proteinuria with impaired renal function was 0.75%. PMID- 2967148 TI - Diabetes in hypertensive women: an effect of antihypertensive drugs or the hypertensive state per se? AB - A total of 226 women participating in a longitudinal study who were found to be hypertensive (initial values greater than or equal to 160 mmHg for the systolic blood pressure and/or greater than 95 mmHg for the diastolic blood pressure) were followed up with respect to the risk of developing diabetes during a 12-year study period. For each year of follow-up, information on whether each participant was taking antihypertensive drugs or not was collected. As a result there were 1339 'treatment years' and 1449 'non-treatment years' of follow-up. All 16 women who developed diabetes during the follow-up period were taking antihypertensive drugs at the time of onset of their diabetes. A similar observation was made when 145 women within the upper age-specific decentiles of systolic blood pressure were followed up in the same way. These results suggest that in hypertensive subjects antihypertensive drugs rather than the hypertensive state per se predispose to diabetes mellitus. PMID- 2967149 TI - Renal artery stenosis in hypertensive diabetic patients. AB - The prevalence of secondary forms of hypertension in diabetes is unknown. One hundred and five of 465 patients randomly selected from a diabetic clinic population were found to be hypertensive. Hypertensive patients aged less than 70 years were screened for renal artery stenosis using intravenous digital subtraction angiography. Two angiograms were technically unsatisfactory. All 18 insulin-dependent patients successfully screened had normal renal arteriograms. Five of 24 non-insulin-dependent patients had unilateral renal artery stenosis but functional tests did not clearly suggest that renal artery stenosis was causing the hypertension in these cases. No patient was referred for surgery or angioplasty. We conclude that renal artery stenosis is common in hypertensive non insulin-dependent diabetics but may not, in many cases, be the cause of the hypertension. The criteria for investigating diabetic hypertensives for renal artery stenosis should be no different from those used in the general hypertensive population. PMID- 2967150 TI - Depression and diabetes. AB - The incidence and prevalence of depression in diabetic patients in the United Kingdom is unknown. Since depression may influence blood glucose control which in turn may be related to the development of diabetic complications, it is important to estimate its prevalence in diabetic patients. The prevalence of depression was investigated in a group of Caucasian and West Indian, insulin-(IDDM) and non insulin-dependent (NIDDM) adult diabetics and a non-diabetic comparison group. Prevalence of depression was 8.5% for both groups and a further 19.2% and 14.6%, respectively, had borderline depression. Presence of depression was unrelated to sex, ethnic group, duration or type (IDDM or NIDDM) of diabetes and social class but significantly related to type of accommodation, marital status, and amount of social contact. A higher percentage of diabetics with psychiatric symptoms had one or more current complications compared to 'normal' diabetics. Diabetics suffer from a similar amount of depression to non-diabetics, but psychiatric symptoms may be related to the frequency of diabetic complications. PMID- 2967151 TI - Management of non-insulin-dependent diabetes mellitus in Europe: a concensus view. AB - Non-insulin-dependent diabetes mellitus (NIDDM) is a very common disorder, but less attention is paid to it than to the more dramatic but less prevalent insulin dependent diabetes. The majority of patients receive their care from primary health care physicians or doctors without particular training in diabetes. Care is highly variable and often inadequate. In the USA this problem has been tackled by promoting a national education programme involving extensive use of television together with written guidelines for management. These latter have been looked at with interest by many diabetologists in Europe, but it has been felt that the guidelines were not entirely suitable for European practice. As a result an initiative was taken to generate more suitable materials for European use. Diabetologists from 14 European countries, representing the European NIDDM Policy Group (EUR-NIDDM-PG) therefore met in Amsterdam in 1986 with the blessing of the Council of the European Region of the IDF. The aims were first to establish whether a concensus view on management of non-insulin-dependent diabetes could be achieved, and if this was possible then to produce written materials for use in the care of people with non-insulin-dependent diabetes aimed particularly at the primary health care sector and the general physician. A remarkable degree of concensus was reached considering the different backgrounds and specialties represented by the group. Discussions were lengthy but constructive and were pervaded by a determination to improve care for such patients in Europe. The first output from the group is the short concensus document which follows.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967152 TI - A passport to improved diabetes care. PMID- 2967153 TI - Ten years experience in a diabetic day centre. PMID- 2967154 TI - Do diabetic children and their parents learn from each other? PMID- 2967155 TI - Antepartum pituitary necrosis in diabetes mellitus. AB - Pituitary infarction occurred during pregnancy in two insulin-dependent diabetic patients aged 32 and 33 years. The diagnosis was delayed in each case before being confirmed biochemically and radiologically. A decreasing insulin requirement and recurrent hypoglycaemia made it possible to determine the exact time that pituitary infarction occurred during pregnancy. The clinical features of these cases should alert physicians to the possibility of pituitary infarction in diabetic pregnancy and minimize the delay in diagnosis. PMID- 2967156 TI - Side-room tests to screen for microalbuminuria in diabetes mellitus. AB - Three side-room tests (latex bead immunoagglutination test, LBT; 25% sulphosalicylic acid test, SST; microalbutest, MAT) for the detection of microalbuminuria in diabetics are described and their screening potential and practicability assessed. One hundred insulin-dependent diabetics attending a diabetic clinic provided an early morning urine sample (Albustix-negative) which was subjected to each of the three tests, and urinary albumin concentration (UA) was assayed by RIA. Tests were assessed in random order by two trained operators using a semiquantitative grading scale with 100% concordance between 10 observers. All test results greater than or equal to trace +ve were sufficiently sensitive (sensitivity greater than or equal to 90%) in detecting UA greater than 15 mg/l, but MAT exhibited a significantly reduced specificity (69%) and positive predictive value (58%). For a reference UA greater than 30 mg/l, LBT and SST results greater than or equal to trace +ve and MAT results greater than or equal to +ve showed a sensitivity of 100%, a specificity greater than 85% and a positive predictive value greater than 60%. Reagent shelf-life was shortest with LBT. SST involved centrifugation or filtration. Technical skill required was highest with LBT and lowest with MAT. Costs were slightly higher with LBT than SST and were not available for MAT. PMID- 2967157 TI - Interaction of gossypol with sperm macromolecules and enzymes. AB - The interaction of gossypol with sperm macromolecules is an important process for the gossypol action. It is of our interests to study the nature of this interaction in order to understand the mechanism of action of gossypol. Our experimental results support the thesis that gossypol inhibits sperm motility by blocking ATP production and utilization. The drug acts on the mitochondria, suppressing oxygen consumption, inhibiting pyruvate dehydrogenase and ATPase activities and probably on the motility apparatus by blocking dynein ATPase activity and preventing protein phosphorylation. The nature of the chemical bond in the interaction of gossypol and macromolecules is through the Schiff base formation and by the hydrophobic and nucleophilic attraction. PMID- 2967158 TI - Retroregulation of bacteriophage lambda int gene expression. PMID- 2967159 TI - Tail length determination in double-stranded DNA bacteriophages. PMID- 2967160 TI - Single-stranded DNA phage origins. PMID- 2967162 TI - Fixed drug eruption due to pseudoephedrine. PMID- 2967161 TI - An unusual cutaneous reaction secondary to allopurinol. AB - A 60-year-old man presented with erythematous papules and plaques on his body as well as a violaceous indurated nodule on his face. A cutaneous drug reaction secondary to allopurinol was considered. Examination of biopsy specimens revealed a dense lymphocytic infiltrate of the skin as is seen in lymphocytoma cutis. The lesions cleared promptly after use of allopurinol was discontinued. A literature review showed that an eruption of this kind, including a nodule on the face that histologically simulated a lymphocytic infiltrate of the skin, has not yet been reported secondary to allopurinol. PMID- 2967163 TI - Polymorphous drug eruption due to nifedipine. AB - A patient who underwent coronary bypass surgery and was treated with nifedipine subsequently developed an erysipelaslike erythematous itching plaque on both shins. Histopathologic examination of a biopsy specimen from the area showed findings compatible with a lichen-planus-like drug eruption. An awareness of varied skin reactions produced by nifedipine may reduce the suffering of patients and help prevent unnecessary local and general treatments. PMID- 2967164 TI - Antitussive properties of inhaled bronchodilators on induced cough. AB - We have studied the antitussive effects of two anticholinergic agents, oxitropium bromide (200 micrograms) and ipratropium bromide (80 micrograms), and a combined beta-agonist and anticholinergic preparation containing fenoterol hydrobromide (200 micrograms) and ipratropium bromide (80 micrograms), in 16 normal and ten asthmatic volunteers in a double-blind, randomized, placebo-controlled crossover trial. Cough was induced by inhalation of ultrasonically nebulized distilled water and hypotonic saline solution. All treatments significantly reduced the cough response to inhaled distilled water aerosol when compared with placebo (p less than 0.001). There was no difference between oxitropium bromide and ipratropium bromide (p greater than 0.05), but the combination preparation displayed a greater antitussive effect than either oxitropium bromide (p less than 0.05) or ipratropium bromide (p less than 0.025). Cough frequencies in response to hypotonic 0.18 and 0.32 percent saline aerosol were lower than those obtained with distilled water (p less than 0.005) for all treatments. Asthmatic patients coughed less frequently than normal volunteers in response to all solutions when placebo was given (p less than 0.05), but there is no evidence to suggest that the response to treatment was different in the two groups. Our results suggest that inhaled anticholinergic bronchodilators alone or in combination with beta 2-adrenergic agonists might be effective in the treatment of pathologic cough. PMID- 2967165 TI - Current management of severe exercise-related cardiac events. AB - Severe cardiac events during either exercise testing or cardiac rehabilitation are described in five patients. The three events related to exercise testing involved patients with atypical chest pain which, in retrospect, probably reflected unstable angina pectoris or coronary vasospasm (or both). Prompt coronary angiography with subsequent percutaneous transluminal coronary angioplasty was performed. Two cardiac arrests (one fatal) occurred during participation in cardiac rehabilitation programs. One was caused by primary ventricular fibrillation. Electrophysiologic testing was used in determining the efficacy of the drug. The fatality was due to an acute myocardial infarction. On site intravenous thrombolytic therapy might prove useful in similar events in the future. Comparisons of surveys published in the 1970s with those in the 1980s show that the mortality and morbidity during exercise testing remain low and that similar rates during cardiac rehabilitation are even lower; however, these cases exemplify that emergency situations can and do arise, necessitating prompt intervention and aggressive subsequent management. PMID- 2967166 TI - [Laparoscopy in children]. PMID- 2967167 TI - Effects of bile salts on plasma concentration of beta-endorphin-like immunoreactivity in men. AB - The effects of bile salts on the release of beta-endorphin-like immunoreactivity (beta-END-LI) were investigated in men using a specific radioimmunoassay developed by the authors. Plasma beta-END-LI was determined after extraction by the acid-acetone method (recovery: 73 +/- 5%). Oral administration of 400 mg of sodium taurocholate caused a rise in plasma beta-END-LI from 9.9 +/- 0.5 pmol/liter to 21.3 +/- 1.2 pmol/liter after 30 min and 18.1 +/- 0.5 pmol/liter after 60 min, with return to the initial value after 90 min. Oral administration of chenodeoxycholic acid (CDCA) also increased plasma beta-END-LI from a basal level of 8.4 +/- 0.7 pmol/liter to 18.7 +/- 0.8 pmol/liter after 30 min. Oral administration of ursodeoxycholic acid (UDCA) increased plasma beta-END-LI from 7.3 +/- 0.3 pmol/liter to 30.6 +/- 0.2 pmol/liter after 30 min. In gel chromatography, the beta-END-LI released after UDCA administration separated into two components, which eluted in the same positions as human beta-lipotropin and human beta-endorphin, respectively. These results suggested that bile salts may participate the release of beta-END-LI. PMID- 2967168 TI - Synteny mapping of the genes for 21 steroid hydroxylase, alpha A crystallin, and class I bovine leukocyte antigen in cattle. AB - Synteny mapping of the genes for bovine alpha A crystallin (CRYA-1), steroid 21 hydroxylase (21OH), and a class I major histocompatibility locus bovine leukocyte antigen (BoLA) was accomplished by Southern blot analysis of DNA from bovine hamster hybrid somatic cells. Comparison of the distribution patterns of CRYA-1, 21OH, and BoLA sequences among the hybrid cells revealed that the genes are located on two different chromosomes in the bovine genome. CRYA-1 mapped with superoxide dismutase (SOD1 in syntenic group U10, while 21OH and BoLA were concordant with glyoxylase (GLO1) in syntenic group U21. Therefore, the syntenic array of CRYA-1, 21OH, and BoLA in cattle is different from that observed in rodents, where the genes are closely linked to each other and to GLO1, and consistent with reported linkages in man. PMID- 2967169 TI - [Background monitoring of mutagenesis in fishery bodies of water]. PMID- 2967170 TI - [High-frequency angioplasty in coronary disease. Initial clinical experiences]. AB - After extensive animal experiments a new method of high-frequency current coronary angioplasty (HFCA) via a specially developed catheter system was used in ten patients with haemodynamically significant coronary artery stenoses. In eight patients the stenoses affected the anterior interventricular branch (AIVB), in one patient each the circumflex branch and the right coronary artery, respectively. In nine patients the stenosis cross-section was reduced by at least 20% (from a mean of 91.9% +/- 5.8% to 52.6 +/- 17.5%). In three patients HFCA was followed by balloon angioplasty. The total number of applications per patient ranged from 3 to 12 (mean total duration of current flow: 3.9 +/- 1.0 s). There was only one severe complication, in a patient with subtotal AIVB occlusion after initially successful HFCA: balloon angioplasty successfully restored the lumen to the post-HFCA state. The new technique of HFCA is worthy of further development. It promises to be a valuable addition or alternative to balloon coronary angioplasty. PMID- 2967171 TI - [Significance of acetylsalicylic acid in the secondary prevention of peripheral arterial occlusive diseases]. PMID- 2967173 TI - [Phototherapy of tumors using photosensitizing agents]. AB - The experimental and clinical data on a new method of cancer treatment- photodestruction of tumours treated with the photosensitizing preparations--are reviewed. Particular emphasis is given to porphyrin compounds. The problems are considered dealing with accumulation of these compounds in cells and tissues, with the mechanism of photoalteration of sensitized tumours, with testing of the method on transplanted and spontaneous animal tumours as well as with the data on the phototherapy of human tumours with a hematoporphyrin-derivative used as a photosensitizer. The prospects for the method development are evaluated. PMID- 2967172 TI - [Morphological correlates of the endocrine function of the heart]. PMID- 2967174 TI - Both type I and II insulin-like growth factor receptor binding increase during lactogenesis in bovine mammary tissue. AB - Bovine GH is a potent stimulant of lactation, and the insulin-like growth factors I and II (IGF-I and -II) are believed to mediate GH's growth-promoting actions. Since all of IGF's known actions are mediated through its receptor subtypes, we analyzed the distribution of IGF receptor subtypes in lactating and nonlactating bovine mammary tissue. Analysis of competition curves showed that IGF-I had greater potency than IGF-II in competing with [125I]IGF-I for binding to membranes prepared from both lactating and nonlactating animals. An insulin concentration of 4 micrograms/ml displaced less than 40% of the [125I]IGF-I bound to membranes prepared from both lactating and nonlactating animals, indicating that a high percentage of [125I]IGF-I was bound to the type II receptor. Lactation was associated with an increase in the total amount of [125I]IGF-I bound, and this change was due to an increase in binding to both receptor subtypes. Specifically, membranes prepared from lactating animals had a 3-fold increase in binding competed for by insulin and a 2-fold increase in binding not competed for by insulin. Affinity cross-linking of [125I]IGF-I to membranes prepared from both lactating and nonlactating animals, followed by polyacrylamide gel electrophoresis (PAGE) and autoradiography, showed that 260K and 135K bands were present. Competition experiments indicated that unlabeled IGF-I effectively competed for binding to the 260K band, whereas insulin did not. Binding to the 135K band could be inhibited by both IGF-I and insulin. The intensity of the labeled bands showed that type II receptors were relatively more abundant than type I receptors in membranes from both lactating and nonlactating animals. Membranes prepared from lactating animals showed both 135K and 127K species of the type I receptor, whereas nonlactating animals showed only the 135K band. We conclude that type I and II receptors are present in bovine mammary tissue, and type II predominate. Lactation is associated with increases in the concentration of both receptor subtypes, especially type I receptors. Lactation may be associated with structural changes in the type I receptor. These changes in receptor distribution could play a role in modulating the physiological effects of the IGFs on mammary tissue. PMID- 2967175 TI - Insulin-like growth factor-binding proteins in hypophysectomized rat liver: characterization and subcellular localization. AB - We have characterized binding proteins for insulin-like growth factors (IGFs) in hepatic subcellular fractions and in the washed supernatants of these fractions in normal and hypophysectomized (hypox) rats. In the course of assessing IGF-II binding sites on rat liver microsomes, we observed that [125I] IGF-II binding to the microsomal membranes of hypox rats was much lower than that in normal rats. Paradoxically, binding increased in hypox animals at low concentrations (0.5-5 ng/ml) of unlabeled IGF-II. After resuspension and centrifugation (washing) of the microsomes, no difference was found in [125I]IGF-II binding to hypox vs. normal microsomes. However, the binding of [125I]IGF-II to the washing supernatant (SN) from hypox rat microsomes was greater than binding to that from normal animals. Binding to SN was inhibited by unlabeled IGF-II in a dose dependent manner. Scatchard analyses indicated that the affinity constant for binding by hypox SN was higher than that of normal SN and the microsomal fractions of both hypox and normal rats. After further subfractionation of the liver, no binding activity was found in SN from plasmalemma, whereas it was about 20% of input counts per min of [125I]IGF-II in SN from combined Golgi-endosome fractions of hypox rat liver. We next compared IGF-binding moieties in microsomal SN with those in plasma using cross-linking of [125I]IGF-II followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In normal rat plasma, we observed the presence of 42K, 39K, 31K, and 27K binding complexes. In hypox rat plasma only a 42-39K doublet was found. In the SN of normal rat microsomes, the predominant complex migrated at 39K and was distinguishable only after acidification. In the SN of hypox rat microsomes, the 42K complex was predominant, with a minor 34K complex. These studies have identified IGF-binding moieties in hepatic tissues, particularly in hepatic vesicular elements, which interfere in the binding of IGF-II to membrane receptors. Their presence in these receptor-rich elements may influence IGF binding to intracellular receptors and, hence, the biological activity of the peptide. PMID- 2967176 TI - Enkephalin convertase in the heart: similar disposition to atrial natriuretic factor. AB - Enkephalin convertase (EC; carboxypeptidase-E or -H) is a carboxypeptidase-B-like enzyme proposed to be involved in the synthesis of enkephalins and other neuropeptides. In this study we have characterized and localized EC in the rat heart and examined its correspondence with atrial natriuretic factor. Heart homogenates bind [3H]guanidinoethylmercaptosuccinic acid ([ 3H]GEMSA), a selective ligand for enkephalin convertase, with specificity and high affinity (Kd = 5-10 nM). The pharmacology of these sites matches that of convertase catalytic activity and of [3H]GEMSA binding in other tissues. The binding sites in the heart can be purified 4000-fold using p-aminobenzoylarginine affinity chromatography, and the purified sites have Co2+ stimulated carboxypeptidase activity identical to EC. By subcellular fractionation studies [3H]GEMSA-binding sites are localized to the granule fraction with ANF immunoreactivity. [3H]GEMSA autoradiography localizes EC in the heart to the left and right atria, with very low levels in mature and ventricles. The distribution, biochemical properties, and developmental course of EC suggest that it may be involved in the synthesis of atrial natriuretic factor. PMID- 2967177 TI - Distribution of insulin-like growth factor II receptor immunoreactivity in rat tissues. AB - Antibodies specific for the insulin-like growth factor II (IGF-II) receptor were used to study its distribution in a number of rat tissues and cell lines in order to determine which cells might be responsive to local or circulating IGF-II. In cultured 18-54,SF and B104 neuroblastoma cells, plasma membrane and cytoplasmic staining corresponding to Golgi apparatus could be seen, consistent with the glycoprotein nature of this receptor. Antibody binding was also seen in the central nervous system, confined primarily to the choroid plexus, and the vascular and ependymal elements. Some staining was seen in the parenchyma of the brain, in addition to binding around nerve sheaths and axon bundles. There were high levels of immunoreactivity in all three lobes of the pituitary, including vascular and cellular elements. In liver, highest levels of immunoreactivity occurred in the sinusoidal cells. In lung, IGF-II receptor immunostaining was seen in the alveoli and around the bronchioles. Staining in kidney was observed in glomeruli, tubules, and Bowman's capsules. Lower levels of immunostaining were seen in skeletal muscle, located primarily around the muscle sheaths. Localization of IGF-II receptor to cells of known function in different tissues will help elucidate the role of this ligand-receptor system in regulating growth and metabolism. PMID- 2967178 TI - Studies on binding and mitogenic effect of insulin and insulin-like growth factor I in glomerular mesangial cells. AB - The mesangial cells, as part of their smooth muscle cell function, are actively involved in regulating glomerular hemodynamics. Their overlying endothelium is fenestrated; therefore, these cells are directly exposed to plasma substances, including hormones such as insulin and insulin-like growth factor I (IGF-I). These peptides may contribute to the mesangial sclerosis and cellular hyperplasia that characterize diabetic glomerulopathy. We report herein the characterization of the receptors and the mitogenic effects of IGF-I and insulin on mouse glomerular mesangial cells in culture. The IGF-I receptor was characterized on intact cells. The Kd of the IGF-I receptor was 1.47 X 10(-9) M, and the estimated number of sites was 64,000 receptors/cell. The binding was time, temperature, and pH dependent, and the receptor showed down-regulation after exposure to serum. The expression of the receptor did not change on cells at different densities. The specific binding for insulin was too low to allow characterization of the insulin receptor on intact cells. However, it was possible to identify the insulin receptor in a wheat germ agglutinin-purified preparation of solubilized mesangial cells. This receptor showed the characteristic features of the insulin receptor, including pH dependence of binding and a curvilinear Scatchard plot. The mitogenic effects of insulin and IGF-I on mesangial cells were measured by the incorporation of [3H]thymidine into DNA. IGF-I was more potent than insulin. The half-maximal response to IGF-I stimulation occurred at 1.3 X 10(-10) M, and a similar increase with insulin was observed at concentrations in the range of 10( 7) M, suggesting that this insulin action was mediated through the IGF-I receptor. These data show that the mouse microvascular smooth muscle cells of the glomerulus express a cell surface receptor for IGF-I in vitro and that this peptide is a potent mitogen for these mesangial cells. It may, therefore, play a role in glomerular proliferative lesions. The insulin receptor is present in small numbers and does not mediate mitogenesis in mesangial cells. PMID- 2967179 TI - Structural determinants of ligand recognition by type I insulin-like growth factor receptors: use of semisynthetic insulin analog probes. AB - We undertook a systematic analysis of the structural determinants necessary for ligand recognition by the type I insulin-like growth factor (IGF) receptor by investigating the binding of semisynthetic insulin analogs to IGF receptors from human placental cell membrane fragments. Analogs were prepared by synthetic and semisynthetic methods. Three groups of insulin analogs were synthesized: the first group contained insulin analogs modified at the amino-terminal position of the insulin A chain and included acetyl-insulin and human proinsulin; the second group included analogs in which B chain residues B26-B30 [despentapeptide insulin (DPI)], B25-B30 (deshexapeptide insulin), and B24-B30 (desheptapeptide insulin) were removed; the third group contained insulin analogs in which B chain residues B26-B30 were removed (DPI) and phenylalanine(B25) substituted with other amino acids, including alanine, serine, leucine, and tyrosine. Half-maximal inhibition of binding of radiolabeled IGF-I to placental cell membrane fragments was used as an index of relative binding affinity (K1/2). To determine further if semisynthetic insulin analogs bound to the type I IGF receptor, placental membrane fragments were affinity labeled with radiolabeled IGF-I in the presence and absence of submaximal concentrations of unlabeled hormone, insulin, or semisynthetic analogs, and the labeled proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Insulin had a 500 times lower affinity for the type I IGF receptor than IGF-I [K1/2 = 140 +/- 69 nM (mean +/- SD)] whereas proinsulin and acetyl insulin had a more than 100 times lower affinity than insulin for this receptor type. Removal of insulin B chain amino acid residues 26-30 (DPI) did not negatively affect the binding of the insulin derived peptide and actually increased the apparent affinity of ligand-receptor association approximately 2-fold. However, further removal of phenylalanine(B25) (deshexapeptide insulin) and phenylalanine(B24) (desheptapeptide insulin) decreased the binding of ligand to the type I IGF receptor progressively by several orders of magnitude. Substitution of phenylalanine(B25) of DPI with tyrosine, a substitution that actually increased the homology of this analog to IGF-I, resulted in a 4- to 5-fold increase in the relative apparent affinity of the analog for the type I IGF receptor (K1/2 = 31 +/- 4 nM). On the other hand, substitution of phenylalanine(B25) with alanine, serine, and leucine decreased the relative apparent binding affinity approximately 2- to 8-fold.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967180 TI - Effects of food deprivation and high fat diet on immunoreactive beta-endorphin levels in brain regions of Zucker rats. AB - The levels of immunoreactive beta-endorphin (ir-beta-EP) were measured in the brain and pituitary of lean Zucker rats subjected to food deprivation for 72 h and to a high fat diet, and in fatty Zucker rats after food deprivation for 72 h. Ir-beta-EP was increased in the neurointermediate (NI-) pituitary lobe but reduced in the medulla-pons of fatty rats when compared to lean littermates fed ad libitum. Food deprivation decreased ir-beta-EP in the cortex and medulla-pons of lean rats and in the cortex, midbrain and NI-pituitary of fatty rats. In contrast, ir-beta-EP was increased in the anterior pituitary of lean rats and in the striatum of fatty rats after deprivation. The high fat diet produced a decrease in ir-beta-EP in the cortex, midbrain and NI-pituitary with an increase in the striatum and hypothalamus of lean rats. These results suggest that the ir beta-EP concentration could be differentially affected in different brain regions of Zucker rats by changes in the energy balance. PMID- 2967181 TI - Isolation and characterization of two proteoheparan sulfate species of calf arterial tissue. AB - When calf aortic tissue, preincubated under organ culture conditions in the presence of [35S]sulfate, was submitted to a sequential collagenase and elastase digestion and guanidinium chloride extraction, the bulk of proteoheparan sulfate was obtained in the elastase fraction. Ion-exchange chromatography on DEAE cellulose of the elastase digest under dissociative conditions yielded a proteoglycan fraction that contained heparan sulfate as the sole glycosaminoglycan. The proteoheparan sulfate fraction was resolved into a high molecular-mass (P-HS 1) and a low-molecular-mass (P-HS 2) fraction by gel filtration on Sephacryl S-400. P-HS 1 has a Mr of 175,000 and possesses four heparan sulfate side-chains (Mr 32,000) covalently bound to the protein core via a galactose- and xylose-containing polysaccharide-protein binding region. The protein core (Mr 38,000), which was obtained after deglycosylation of PG-HS 1 with trifluormethane sulfonic acid, contained in addition a few N-glycosidically linked oligosaccharide units representing a complex type with terminal neuraminic acid residues. P-HS 2 is a single-chain peptidoheparan sulfate of Mr of 38,000 containing one heparan sulfate chain (Mr 32,000) linked to a polypeptide (Mr 6000). The ratio of specific radioactivities of P-HS 1 and P-HS 2 was 1:0.66. PMID- 2967182 TI - Polymerization of actin modified with fluorescein isothiocyanate. AB - Solution properties of skeletal muscle actin, modified at lysine-61 with fluorescein isothiocyanate (FITC) [Burtnick, L.D. (1984) Biochim. Biophys. Acta 791, 57-62], were re-examined in this work by light scattering, analytical ultracentrifugation, fluorescence, electron microscopy and myosin ATPase activity measurements. Fluorescence measurements using trace amounts of actin labeled with N-(1-pyrenyl)iodoacetamide showed that the FITC modification inhibited but did not block completely the polymerization of actin by KCl and MgCl2. Sedimentation velocity runs of FITC-actin, incubated with 100 mM KCl and 2 mM MgCl2, revealed the presence in these solutions of polymeric, oligomeric and monomeric species. The critical concentration for FITC-actin polymerization under these conditions was 12 microM. As judged by electron microscopy, FITC-actin polymers were similar to but generally shorter than standard F-actin filaments. Light scattering measurements indicated that FITC modification inhibited also the polymerization of actin by myosin subfragment 1 (S1) but the resulting complexes were indistinguishable from standard, decorated actin filaments. MgATPase measurements showed that FITC-actin, polymerized by preincubation with S1, activated the MgATPase activity of S1 while the monomeric labeled protein did not. Thus, in analogy to native actin, the activating function of FITC-actin depended on the formation of actin filaments. Results presented in this study suggest that the region around lysine-61 of actin plays an important role in actin-actin contact and is less crucial to actomyosin interaction. PMID- 2967183 TI - Exercise blood pressure and heart rate reduction 24 and 3 hours after drug intake in hypertensive patients following 4 weeks of treatment with bisoprolol and metoprolol: a randomized multicentre double-blind study (BISOMET). AB - In a 4-week randomized, double-blind study, 87 patients with essential hypertension received either 10 mg bisoprolol (B) or 100 mg metoprolol (M) once daily (o.d.). The effects of the beta blockers on systolic blood pressure, heart rate and rate-pressure product during exercise, 24 h (E2) and 3 h (E3) after administration (p.a.) were compared with the values obtained in the baseline exercise test (E1). 24 hours p.a. the effects of B were significantly stronger than of M (E1-E2: B vs M; P less than 0.01) whereas 3 h p.a. no significant differences were detectable between B and M. The residual effects 24 h p.a. in relation to the effects 3 h p.a. (E1-E2/E1-E3) were significantly greater with B (86-93%) than with M (53-66%). In contrast to the findings with 100 mg M o.d., 10 mg bisoprolol o.d. guarantees a persistent reduction in exercise blood pressure and heart rate throughout the entire dosage interval of 24 h. PMID- 2967184 TI - A comparison of once daily bisoprolol, 5 and 10 mg, and atenolol 100 mg in the treatment of angina pectoris. AB - The efficacy and safety of the new cardioselective beta-blocker bisoprolol 5 mg and 10 mg once daily in the treatment of angina pectoris was compared with atenolol 100 mg once daily. 19 patients with coronary artery disease and angina of effort completed a randomised, three-way crossover study which consisted of six-week treatment periods preceeded by a two-week placebo washout. Treadmill exercise stress tests were carried out approximately 24 hours after each dose, at the end of the placebo period and after 2 and 6 weeks of each active treatment period. Patients recorded their angina attack rate and GTN consumption on diary cards. Number of anginal attacks, GTN consumption, resting and exercise systolic blood pressure, heart rate and double product were reduced, and both exercise duration and time to 1 mm ST-segment depression during exercise were increased to a similar extent by all three treatment regimens (P less than 0.01, paired t test). The only differences between the treatments were that the reductions in heart rate, double product and exercise blood pressure tended to be smaller after 6 weeks of bisoprolol 5 mg. Generally, there was no significant difference between the improvements measured at week 2 of each phase and at week 6 (P greater than 0.05, analysis of variance). All three dosage regimens were well tolerated. The data indicate that there is no difference between the safety and efficacy of bisoprolol 10 mg and atenolol 100 mg once daily in the treatment of angina pectoris. In addition, the 10 mg dose of bisoprolol would seem to have only a small advantage over the lower 5 mg once daily dose. PMID- 2967185 TI - Bisoprolol in the treatment of angina pectoris: a double blind comparison with verapamil. AB - In order to verify the anti-ischaemic effect of a new beta-blocking agent, bisoprolol, a double blind parallel groups trial was carried out in comparison with verapamil. 26 patients with a history of spontaneous and/or effort angina were studied. After a two-week treatment with placebo, they were randomized in two groups. One group was treated for 4 weeks with bisoprolol 10 mg o.d. and for the following 4 weeks with bisoprolol 20 mg o.d. The other group received verapamil 80 mg t.i.d. for the first 4 weeks and 120 mg t.i.d. for the remaining 4 weeks. Throughout the study isosorbide dinitrate 20 mg b.i.d. was administered and sublingual nitroglycerin was allowed when necessary. 21 patients completed the study. Both bisoprolol and verapamil significantly reduced the number of angina episodes and nitroglycerin tablets consumption, as well as ischaemic episodes recorded on Holter ECG. The total number and severity of ectopic ventricular beats were reduced too. On multistage treadmill exercise test, both drugs increased effort time and time to ST depression = 1 mm, and reduced ST depression and double-product. The effect of bisoprolol on double product was greater than that of verapamil because of the better control of heart rate. The relationship ST/double product suggested that beta-blockers act essentially through the reduction of myocardial oxygen consumption and verapamil possibly with an additive effect on coronary circulation. Radionuclide ventriculography showed no deterioration of rest ventricular function with both drugs. In conclusion, bisoprolol and verapamil showed a satisfactory anti-ischaemic effect, with good tolerability. PMID- 2967186 TI - Efficacy and safety of bisoprolol and atenolol in patients with mild to moderate hypertension: a double-blind, parallel group international multicentre study. AB - Three hundred and fifteen patients were randomly allocated to treatment for six months with bisoprolol 5 or 10 mg day-1 or atenolol, 50 mg day-1, in a double blind, double-dummy parallel group, international multicentre study. Two hundred and ninety-two (175 men and 117 women) were eligible for statistical follow-up. Their mean age was 52.6 years (range 28-70). All patients had a supine diastolic blood pressure of 95-120 mmHg on two occasions during the four weeks of placebo treatment. Twenty-four patients ended the study prematurely and a further 19 had their regimes changed because of an insufficient effect. The reasons for drop-out were similar in the three treatment groups. Thus, 249 patients continued to receive the treatment they were allocated to, with 80, 83 and 86 patients in the three respective groups. The sex and age distributions and the number of previously treated hypertensives were similar in the three groups. At the end of placebo treatment the supine blood pressures in the three groups (bisoprolol 5 or 10 mg day-1 or atenolol 50 mg day-1, respectively) were 163.9/102.5, 157.4/101.8 and 160.0/102.2 mmHg, respectively. The systolic blood pressure was higher (P less than 0.05) in the group receiving bisoprolol 5 mg day-1 than in the 10 mg day-1 group. After 26 weeks of treatment the supine blood pressures in the three groups were 150.6/90.8, 142.0/89.1 and 148.6/91.7 mmHg, respectively. The largest estimated difference in blood pressure reduction was 4.6/2.3 mmHg between the group receiving bisoprolol 10 mg day-1 and the group receiving atenolol 50 mg day 1. A reduction in mean blood pressure of greater than or equal to 10 mmHg was noted in 66% of the patients in the bisoprolol group (10 mg day-1), in the other groups 66 and 59%, n.s. Bisoprolol is effective, well-tolerated and safe in the treatment of hypertension. A daily dose of 5 mg seems recommendable for the majority of hypertensive patients and seems equipotent with 50 mg day-1 of atenolol in the present study. PMID- 2967187 TI - Comparison of bisoprolol with nifedipine for treatment of essential hypertension in the elderly: comparative double-blind trial. AB - Beta blockers are often considered to be less effective for blood pressure control in elderly patients with essential hypertension than calcium antagonists. We therefore compared the efficacy and tolerability of the new beta blocker bisoprolol with those of nifedipine. Fifty-nine patients over 60 years of age with essential hypertension (supine diastolic blood pressure, DBP: 95-115 mmHg) took part in a randomized double-blind study. After 14 days on placebo the patients received either 10 mg bisoprolol o.d. or 20 mg nifedipine SR b.i.d. for 4 weeks. The doses were doubled in patients whose DBP remained greater than 90 mmHg, for a further 4 week period. Blood pressure was measured about 24 hours after bisoprolol and about 12 hours after nifedipine administration. Fifty-six patients were available for efficacy analysis. After 4 weeks there was a distinct decrease of SBP and DBP in both groups (bisoprolol: -22 +/- 16; -15 +/- 9 mmHg; nifedipine: -24 +/- 17; -17 +/- 7 mmHg) with no significant difference between the treatments. DBP was normalized in 22/28 (79%) patients on bisoprolol and in 24/28 (86%) patients on nifedipine (bisoprolol vs nifedipine: NS). There were 7/29 bisoprolol and 14/30 nifedipine patients who experienced at least one side effect. Overall the tolerability of bisoprolol was considered to be better than that of nifedipine (P = 0.043). In conclusion, bisoprolol 10-20 mg o.d. proved to be as equally effective as nifedipine SR 20-40 mg b.i.d. for the treatment of essential hypertension in the elderly. PMID- 2967188 TI - Food intake, neuroendocrine and temperature effects of 8-OHDPAT in the rat. AB - Administration of 8-hydroxy-2(di-n-propylamino)tetralin (8-OHDPAT) to rats produced dose-dependent decreases in food intake and hypothermia, increases in plasma prolactin and corticosterone, and a decrease in plasma growth hormone. 8 OHDPAT administration also induced the serotonin behavioral syndrome at all doses. Pretreatment with metergoline did not affect the 8-OHDPAT-induced behavioral syndrome or decrease in food intake but attenuated the prolactin increase and, furthermore, potentiated 8-OHDPAT-induced hypothermia. Pretreatment with ritanserin or naloxone did not modify 8-OHDPAT-induced changes in food intake, temperature or prolactin. Similarly, pretreatment with phenoxybenzamine, propranolol, clonidine, haloperidol and methiothepin also did not attenuate 8 OHDPAT-induced decreases in food intake. Administration of pindolol alone produced hyperthermia, decreased food intake and enhanced prolactin secretion. Pindolol thus appears to act as a partial 5-HT agonist in addition to being an antagonist at central 5-HT receptors. PMID- 2967189 TI - Hyperinsulinemia of the genetically obese (fa/fa) rat is decreased by a low dose of the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH DPAT). AB - Changes in glycemia and insulinemia were determined in conscious lean (FA/?) and obese (fa/fa) rats after acute administration of the 5-HT1A receptor agonist 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). The intravenous injection of a low dose of 8-OH-DPAT (150 micrograms/kg) to lean rats rapidly promoted hyperglycemia. This modification was associated with a slight increase in insulinemia. The injection of 8-OH-DPAT markedly decreased basal hyperinsulinemia in obese rats while inducing hyperglycemia. Further evidence of the strong inhibitory effect of 8-OH-DPAT on insulin release was obtained in lean and obese rats during glucose tolerance tests. Intracerebroventricular injection of 8-OH DPAT (45 micrograms/animal) triggered hyperglycemia and markedly decreased insulinemia in both lean and obese rats. This hypoinsulinemic effect of 8-OH-DPAT was more pronounced in the obese than in the lean animals. Measurement of the food intake elicited by 8-OH-DPAT (500 micrograms/kg s.c.) showed that the hyperphagic action of the 5-HT1A agonist was the same in FA/? and fa/fa rats. It is suggested that: (i) hyperinsulinemia of the genetically obese rat may be diminished by a low dose of 8-OH-DPAT; (ii) 5-HT1A autoreceptor-mediated regulation of serotonergic activity is not different in lean (FA/?) and obese (fa/fa) rats; (iii) 8-OH-DPAT could be of potential therapeutic use for some aspects of the pathology of type II diabetes. PMID- 2967190 TI - Structure-activity relationships of alpha-human atrial natriuretic peptide. AB - The spasmolytic activity of synthetic alpha-human atrial natriuretic peptide (alpha-hANP) and its related peptides was determined in vitro using the chick rectum and the rat aorta. Natriuretic activity was also measured in the anesthetized rat, alpha-hANP-(7-28), with the NH2-terminal hexapeptide truncated, had greater spasmolytic and natriuretic effect than did alpha-hANP-(1-28). These responses were reduced by truncation of the COOH-terminal residues. alpha-hANP-(7 23), the cyclic structure of alpha-hANP-(1-28), exhibited weak aortic relaxation and natriuretic activities. However, alpha-hANP-(7-23) produced a greater relaxation than did alpha-hANP-(1-28) in the chick rectum. Elimination of Gly at position 9 reduced the spasmolytic and natriuretic activity. Substitution of amino acid residues at position 8, 12 and 13 changed these activities. Analogues containing the ethylene linkage instead of the disulphide bond had weak biological activity. These results indicate that the size of the 17-amino acid ring and the COOH-terminal residues of alpha-hANP are important for the expression of spasmolytic and natriuretic activity. PMID- 2967191 TI - Gonadotrophin- and androgen precursor-stimulated testosterone secretion by interstitial cells from Mongolian gerbil testes: influence of plasma proteins and elevated temperature. AB - Interstitial cells isolated from Mongolian gerbil testes have been used to investigate the effects of plasma proteins and incubation temperature on HCG- and androgen precursor-stimulated testosterone secretion. Short term (15 min) incubation of interstitial cells with various precursors resulted in a significant increase of testosterone release. On the other hand, no stimulatory effect of HCG (10 mIU) could be observed. Precursor (e.g. progesterone) stimulated testosterone secretion linearly increased with cell concentrations (0.5 x 10(5) to 4.0 x 10(5) cells/0.7 ml medium, r = less than 0.99, p less than 0.001). In the presence of 50% horse plasma, progesterone-stimulated testosterone secretion was even more pronounced. Similarly, also gerbil, rat, calf or human plasma significantly increased progesterone-stimulated testosterone output. Interestingly, this effect was markedly reduced in the presence of cortisol. While incubation of interstitial cells for 15 min at either 40 degrees C or 42 degrees C had no significant effect on androgen precursor- or HCG-stimulated testosterone secretion, incubation of cells at 44 degrees C resulted in a drastic reduction of HCG-stimulated testosterone release, without affecting progesterone- or DHEA-stimulated testosterone secretion. Taken the simplicity to make interstitial cells unresponsive to HCG into account, heat-treated cells might prove to be a versatile tool to distinguish between HCG- and protein-/androgen precursor-stimulated testosterone secretion in vitro. PMID- 2967193 TI - Laparoscopic and transvaginal ova recovery: the effect on ova quality. AB - Transvaginal follicle aspiration guided by transvaginal ultrasound for ova recovery is rapidly gaining popularity in many centers practicing in vitro fertilization and embryo transfer (IVF-ET). Cycle outcome following this new method has not been directly compared to the traditional, laparoscopic recovery technique. To this end, the authors evaluated multiple parameters in 66 laparoscopic (group A), and 44 transvaginal ova recovery procedures (group B) in patients undergoing IVF-ET. No statistically significant differences could be demonstrated between the groups in all but the rate of ova fertilization. The rate of fertilization was higher in the ova recovered by transvaginal follicle aspiration (59.6 versus 69.2%; P less than 0.01). No difference could be demonstrated between the groups in the other parameters examined, which included the number of ova recovered (5.7 +/- 0.4 versus 6.0 +/- 0.7), ova maturity (87 versus 84% intermediate ova), rate of polyspermic fertilization (3.9 versus 5%), rate of cleavage (88 versus 91%), cleavage stage at transfer (3.7 +/- 0.8 versus 3.5 +/- 0.4 cells per embryo), number of embryos transferred per patient (2.7 +/- 0.1 versus 3.3 +/- 0.2), and pregnancy rates. The potential detrimental effects of general anesthesia and CO2 pneumoperitoneum present during laparoscopy but not ultrasound guided recovery on ova quality may underlie the observed difference in fertilization between the groups. PMID- 2967192 TI - The effect of an acute maternal stress on beta-endorphin and growth hormone releasing factor in the rat fetus. AB - Pregnant rats were injected with saline or L-tyrosine methylester HCl (200 mg/kg) and subjected to an acute forced immobilization stress on day 20 of gestation. At 10, 30, 60, and 120 minutes after the onset of stress, their fetuses were dissected out, and the contents of hypothalamic and pituitary immunoreactive beta endorphin (IR-beta-EP) and hypothalamic immunoreactive growth hormone-releasing factor (IR-GRF) were determined by specific radioimmunoassays. The maternal stress arose a significant decrease of hypothalamic IR-beta-EP at 30 minutes, while pituitary IR-beta-EP slightly elevated at 30 minutes, then declined at 60 minutes. Hypothalamic IR-GRF showed a gradual increase during the maternal stress. Tyrosine supplementation tended to attenuate stress-induced changes in hypothalamic and pituitary IR-beta-EP, but the response of hypothalamic IR-GRF was less modified by tyrosine. These results showed the functional changes in fetal central beta-EP and GRF under maternal stress in the late gestational life, and suggested that catecholaminergic regulations participate, at least in part, in the fetal neuroendocrine response to maternal stress. PMID- 2967194 TI - Salpingoscopy: an adjunctive technique for evaluation of the fallopian tube. AB - The success of tubal surgery is dependent upon both the nature and extent of tubal damage as well as the expertise of the surgeon. The present study describes the new technique of salpingoscopy to further appraise the status of tubal health by examination of the endosalpinx, utilizing readily available equipment. There were no complications during salpingoscopy attributable to the endoscopic procedure itself, except for a minimal amount of bleeding occasionally noted on the tubal serosa at the site of forceps stabilization. A discordance of 23.5% was noted overall between the fimbrial appearance at surgery and salpingoscopic examination. This new technique of tubal evaluation may eventually help the reproductive surgeon to more accurately counsel the patient in the choice between tubal reconstructive surgery or IVF-ET, one of the most difficult decisions in reproductive surgery today. PMID- 2967195 TI - Doppler assessment of uterine blood flow changes in patients with fibroids receiving the gonadotropin-releasing hormone agonist Buserelin. AB - Uterine arterial blood FVWs were studied by doppler ultrasound in eight patients with large uterine fibroids who were treated with the GnRH agonist Buserelin intranasally for 4 months. At the end of treatment, reductions in the uterine and discrete fibroid volumes were associated with significant increases in the mean RI values for the uterine (P less than 0.01) and fibroid (P less than 0.001) vasculature, thus indicating reduction in blood flow. PMID- 2967196 TI - Gonadal steroids modulate human monocyte interleukin-1 (IL-1) activity. AB - Interleukin-1 (IL-1), a monocyte and macrophage product, is the mediator of a wide variety of immune responses, including fever, and increases during the luteal phase of the menstrual cycle, concomitantly with progesterone (P). These studies were undertaken to assess the possible relationship between steroidogenesis and IL-1 activity elaborated by human peripheral monocytes. IL-1 was measured with the D-10 thymocyte stimulation bioassay and dose response curves as a function of both estradiol (E2) and P were established. Low concentrations of both E2 (10(-9) M-10(-10) M) and P (10(-8) M-10(-9) M) resulted in maximal IL-1 stimulation. At higher concentrations of both E2 (10(-7) M) and P (10(-7) M-10(-5) M) there was a significant reduction (P less than 0.05) in IL-1 activity. However, neither E2 nor P production by human granulosa-luteal cells cultured with increasing amounts of human IL-1 (0 to 20 IU/ml) was affected in either the absence or presence of human chorionic gonadotropin (hCG, 5 IU/ml). Thus, human peripheral monocyte IL-1 activity appears to be modulated by gonadal steroids; however, a reciprocal relationship between IL-1 and gonadal steroidogenesis was not observed. PMID- 2967198 TI - Access through mobility. PMID- 2967197 TI - [The contractile function and calcium-transport system of the myocardium in aging]. AB - Experiments with different species of animals and different models (in situ heart, isolated perfused heart, isolated papillary muscle) revealed a reduction of functional capacity of ageing heart. A decrease of Ca2+ uptake by sarcoplasmic reticulum and mitochondria and diverse age-dependent shifts were found in sarcolemma. The data obtained suggest that the development of the cardiac contractile function disorders in ageing largely depends of the age-related changes in the Ca2+ transport system. PMID- 2967199 TI - AIDS and dentistry: the legal and ethical issues. A DM conference. PMID- 2967200 TI - AIDS: what dentists are doing about it. PMID- 2967201 TI - The dental hygienist's role in a pediatric oncology center. PMID- 2967202 TI - The subject of AIDS. PMID- 2967204 TI - Teamwork--a different approach. PMID- 2967203 TI - An approach to the dental care of handicapped adults: the hygienist's role. PMID- 2967205 TI - Dental care for the mentally handicapped--a duty or a pleasure? PMID- 2967206 TI - Reports toxic reaction after four years of glove use. PMID- 2967207 TI - Finding dependable disability insurance. PMID- 2967208 TI - There are alternatives to non-allergenic gloves. PMID- 2967209 TI - Fixed drug eruption due to chlormezanone. PMID- 2967210 TI - Fixed drug eruption in an 8-month-old infant. PMID- 2967211 TI - Mechanism of inhibition of beta-hexosaminidase B from human liver by short- and medium-chain monocarboxylic acids. AB - Short- and medium-chain monocarboxylic acids showed an inhibitory effect on enzymatic activity of beta-hexosaminidase B (Hex B) when 4-methylumbelliferyl-2 acetamido-2-deoxyglucopyranoside (MU-GlcNAc) was used as substrate: 1. Two groups were distinguished according to the chain length of the monocarboxylic acids: the first was only constituted by acetic acid (C2) whereas the second group exhibited a broader chain length specificity for medium-chain monocarboxylic acids (between C6 and C9). 2. Both groups were reversible competitive inhibitors (Km = 0.52 +/- 0.15 mM; KiC2 = 21.4 +/- 3.0 mM; KiC7 = 3.4 +/- 0.5 mM). Competition experiments between C2 and C7 (as representent of medium-chain monocarboxylic acids group) demonstrated that these inhibitors were bound to different subsites. 3. Competition experiments between C2 and 2-acetamido-2-deoxy-D-galactonolactone (GalNAcLone) (a competitive inhibitor of lysosomal hexosaminidases) demonstrated that these two inhibitors were mutually exclusive, i.e. they were probably bound at the same subsite. This feature and the structural analogy of C2 with the acetyl residue of GalNAcLone (and of the saccharidic part of the substrate) suggested that C2 bound to the substrate site where the N-acetyl residue of the beta-N-acetyl hexosaminide was positioned. 4. The inhibitory effect of medium chain monocarboxylic acids (C6 and C9) was dependent on their physical state. Below the critical micellar concentration (CMC), detected by a dye spectral shift method, no significant inhibition was detected, but as extensively reported using C7, an obvious inhibitory effect occurred at concentrations higher than CMC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967212 TI - Aggregate formation from thermolabile intermediates in the maturation of the thermostable P22 tailspike. PMID- 2967213 TI - Characterization of the insulin-like growth factor (IGF) receptor in K562 erythroleukemia cells; evidence for a biological function for the type II IGF receptor. AB - Erythroleukemia cells (K562) were found to bind insulin-like growth factor-II (IGF-II) about 10 times as much as IGF-I, insulin and human growth hormone. The specific binding of IGF-II increased as a function of cell number in a range of 0.5-6 X 10(6) cell/ml. Kinetic studies revealed that binding was time and temperature dependent and showed a broad pH optimum. Specificity studies showed no inhibition of 125I-IGF-II binding by insulin or unrelated peptide hormones. The half-maximal inhibition of 125I-IGF-II binding to K562 cells was achieved at 87 and 28 ng/ml of IGF-I and IGF-II respectively. However, the addition of 7.5 and 1.7 ng/ml of unlabeled IGF-I and IGF-II respectively increased the binding of 125I-IGF-II by 55%. This 'hook' effect was greatly reduced when K562 cell membranes were used. Scatchard analysis of IGF-II binding showed a comparable equilibrium constant with either intact cells (Ka = 8.9 X 10(8) M-1) or microsomal membranes (Ka = 5.4 X 10(8) M-1). Cross-linking studies indicated that both 125I-IGF-II and 125I-IGF-I bound to an entity of 215 kDa which increased to 260 kDa under reducing conditions. Both IGF-I and II stimulated 3H-thymidine incorporation into K562 cells whereas insulin was without effect. These data show that both IGF-I and II bind predominantly to a type II-IGF receptor in K562 cells. Since both peptides stimulate 3H-thymidine incorporation in these cells it is possible that the type II-IGF receptor is mediating an anabolic biological response in these cells. PMID- 2967214 TI - Diabetes prevention in BB rats by frequent blood withdrawal started at a young age. AB - The BB rat diabetic syndrome has been prevented by various immunosuppressive and reconstitution measures. We observed an effect of multiple blood samplings on diabetes incidence and examined its immunological correlates. Individual litters were divided into two groups; one was sampled and the other was sham sampled as the control group. Sixty-four diabetes-prone and 59 non-diabetes-prone rats were studied. The sampled rats had blood removed at 15 (28% of total blood volume), 30 (30%), 50 (21%), 75 (16%), and 120 days of age. The sham-sampled control rats had blood removed only at 120 days of age. The incidence of diabetes in the sampled group was markedly lower than that of their sham-sampled littermates (22 vs. 78%). This result was associated with a correction of their OX19+ (pan-T lymphocytes) and W3/25+ (helper/inducer) T-lymphocyte-number defects. An increase in lymphocyte subsets was also seen in the non-diabetes-prone BB rats, significant for all but the OX19+ cells. Islet pathology and pancreatic insulin content were consistent with metabolic outcomes. The effect of blood withdrawal thus has implications for understanding the pathogenesis of both the diabetes syndrome and the lymphopenia of the BB rat. Furthermore, it suggests that a stimulation of lymphopoiesis by blood withdrawal (analogous to that of erythropoiesis) may be a hitherto unrecognized physiological response in normal animals. PMID- 2967215 TI - Increased activity of intestinal acyl-CoA: cholesterol acyltransferase in rats with streptozocin-induced diabetes and restoration by insulin supplementation. AB - We examined the activities of intestinal acyl-CoA:cholesterol acyltransferase (ACAT) and cholesterol esterase, enzymes regulating cholesterol absorption, in rats with streptozocin-induced diabetes (STZ-D) to clarify the effect of diabetes on cholesterol absorption. Three weeks after the induction of diabetes, plasma cholesterol levels were slightly but significantly increased in diabetic rats compared with control animals, whereas a far more remarkable increase in plasma cholesterol was observed in diabetic rats when fed an atherogenic diet containing 1% cholesterol, 0.5% cholic acid, and 5% lard. Microsomal ACAT activity in intestinal mucosa was three times higher in diabetic than in control rats. However, no significant difference in the enzyme activity could be detected between diabetic animals fed control chow and those fed the atherogenic diet. Furthermore, insulin supplementation given to diabetic rats caused a reduction of enzyme activity to the levels found in control animals. In contrast, cholesterol esterase activity in rat intestinal mucosa was unaffected by either the induction of diabetes or the atherogenic diet feeding. In conclusion, we disclosed that apparent ACAT activity in intestinal mucosa is elevated in STZ-D rats. Therefore, we postulate that enhancement of CoA-dependent cholesterol esterification in the intestine might be one of the major factors responsible for hypercholesterolemia in diabetes. PMID- 2967216 TI - Disability in the Third World. PMID- 2967218 TI - More on Doppler flowmetry and portal hypertension. PMID- 2967217 TI - Culture of mouse pancreatic islets in different glucose concentrations modifies B cell sensitivity to streptozotocin. AB - There have previously been divergent data published regarding the effects of glucose on the diabetogenic effects of streptozotocin. In order to further explore this issue, two separate sets of experiments were performed. In the first, mouse pancreatic islets were maintained in culture for 3 days at different glucose concentrations (5.6, 11.1 and 28 mmol/l) and then exposed to streptozotocin. After another 3 days in culture at 11.1 mmol/l glucose, the B cell function was evaluated by measurement of glucose-stimulated insulin release, the number of islets recovered after culture, and the islet DNA and insulin contents. In the second group of experiments islets were first maintained in culture at 11.1 mmol/l glucose, then treated with streptozotocin and subsequently cultured for 6 days at the different glucose concentrations given above. It was found that islets maintained in a medium containing 28 mmol/l glucose before or after streptozotocin exposure showed less signs of damage than islets cultured in 11.1 mmol/l glucose. A similar, but less pronounced, decreased sensitivity to streptozotocin was found in islets precultured in 5.6 mmol/l glucose, in comparison with those islets cultured in 11.1 mmol/l glucose. Culture at 5.6 mmol/l glucose just after streptozotocin treatment did not induce any improvement in islet survival or function. It is suggested that the increased damage induced by streptozotocin to islets precultured at 11.1 mmol/l glucose, in comparison with 5.6 mmol/l glucose, can be related to the fact that an increased metabolic activity of B cells render them more susceptible to the toxin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967219 TI - [Beta-endorphin in premature labor and in mature newborn infants following vaginal and abdominal delivery]. AB - Umbilical venous plasma concentrations of beta-endorphin in 20 premature and 30 mature newborn infants after vaginal delivery or elective caesarean section were determined by specific RIA. In the premature infants the beta-endorphin levels after vaginal and abdominal delivery were significantly higher than in mature newborns. Our data indicate that for premature infants the delivery is more stressful and therefore most likely the release of beta-endorphin from the pituitary is more pronounced. Since high opioid levels can cause respiratory and circulatory difficulties, premature infants are exposed to risk of these problems. PMID- 2967220 TI - [Pelviscopic adhesiolysis. Successes in the treatment of chronic abdominal pain caused by adhesions in the lower and middle abdomen]. AB - Inspecting 200 patients with abdominal pain pelviscopically, we found adhesions and no other pathology in 62 (32%). Three groups were defined according to the extent of the adhesions. In 52 patients (84%), complete adhesiolysis was achieved pelviscopically. At discharge 3-5 days post-operatively, 38 of these patients (73%) were free from complaints; in 15%, the pain had lessened. 6 months after pelviscopy, the patients were again asked about their complaints. 39 (75%) of the patients with complete adhesiolysis answered the questionnaire. 15 (38%) were completely free from complaints after 6 months; in 8 others, the pain had lessened; in 60% of them, the abdominal pain had lasted for more than 6 months. Our results justify an attempt at complete adhesiolysis in all patients with abdominal pain, if adhesions are detected pelviscopically. PMID- 2967221 TI - [Postnatal diagnosis of the most important hemoglobinopathies and erythrocyte enzymopathies]. PMID- 2967222 TI - [Role of predisposing factors in the development of hypoplastic anemia]. PMID- 2967223 TI - [Hygienic aspects of work arrangements for persons with limited work capacity in modern poultry plants]. PMID- 2967224 TI - [History of the creation of a health organization in Nizhnii Novgorod (the city of Gorki)]. PMID- 2967225 TI - [25th anniversary of the Department of Radiation Hygiene of the Leningrad Kirov Institute of Advanced Training of Physicians]. PMID- 2967226 TI - [Incidence of nonspecific lung diseases among rural and urban populations]. PMID- 2967227 TI - [Epidemiological research on the late sequelae of exposure to occupational factors]. PMID- 2967228 TI - [Industrial hygiene in the fabrication of ceramic tile from the wastes of picture tube manufacture]. PMID- 2967229 TI - [Working conditions in the isolation of heavy and rare metals from the wastes of copper smelting production]. PMID- 2967230 TI - "I won't accept AIDS patients...". PMID- 2967231 TI - In vitro growth regulation of endometrial carcinoma cells by tamoxifen and medroxyprogesterone acetate. AB - The growth inhibitory effects of medroxyprogesterone acetate (MPA) and tamoxifen (TAM) were tested on three long-established endometrial carcinoma cell lines (HEC 1, KLE, and RL95-2) and on UM-EC-1, a new endometrial carcinoma cell line established in our laboratory. MPA and TAM were used in growth experiments either alone, simultaneously, or sequentially. The MCF-7 breast cancer cell line was used as a control. None of the endometrial carcinoma cell lines showed significant sensitivity to 0.1-10 microM MPA. In contrast, 10 days exposure to 5 microM TAM induced 83 and 70% growth inhibition in HEC-1 and KLE cultures, whereas the growth of UM-EC-1 was inhibited by 99.7% and RL95-2 cultures by 100%. TAM-induced growth inhibition was reversible since all cell lines resumed logarithmic growth when TAM was removed from the culture medium. Addition of 17 beta-estradiol (E2) to the culture medium did not accelerate recovery, and reversal of TAM-induced growth inhibition was not seen when TAM and E2 were added simultaneously. This is consistent with our finding that, except for MCF-7, these cell lines did not show detectable estrogen receptor (ER) activity in assays performed at the time of these experiments. When treated sequentially with TAM and MPA, all cell lines resumed logarithmic growth when medium containing TAM was replaced with medium containing MPA. Simultaneous exposure to 5 microM MPA and 5 microM TAM resulted in a slight additive growth inhibitory effects only in KLE cultures. Our results show that MPA does not have growth inhibitory effects in these endometrial carcinoma cell cultures, whereas TAM exerts a potent inhibitory effect that is not reversed by estrogen and may thus be mediated through a mechanism different from blockade of ER. In vitro results with the UM-EC-1 cell line correlated with the clinical response of the cell line donor. Her disease progressed during postoperative MPA therapy, but subsequently she responded to TAM therapy. PMID- 2967232 TI - Trace elements during 2 years' oral contraception with low-estrogen preparations. AB - In 17 healthy women, taking either a combination of 0.030 mg of ethinyl estradiol and 0.150 mg of levonorgestrel (n = 9) or a combination of 0.030 mg of ethinyl estradiol and 0.150 mg of desogestrel (n = 8) for oral contraception, the profiles of iron, calcium, copper, and zinc levels were investigated. The blood samplings were performed prior to oral contraception and after 3, 12, and 24 months' contraception, as well as within 2 months after discontinuation of the pill. No changes occurred in iron, calcium, and zinc levels. On the other hand, the copper level was significantly increased during oral contraception, yet returned to the initial level after discontinuation of contraception. No differences occurred between the two preparations for oral contraception. PMID- 2967233 TI - Progestagen-dependent effect on some plasma proteins during oral contraception. AB - Seventeen healthy women received a combination of 0.030 mg of ethinyl estradiol and 0.150 mg of levonorgestrel or a combination of 0.030 mg of ethinyl estradiol and 0.150 mg of desogestrel for 2 years as oral contraception. Serum levels of sex hormone binding globulin, transcortin, ceruloplasmin, and pregnancy associated protein were measured before contraception, during 3, 6, 12, 18, and 24 months of treatment, and 2 months after stopping the pill. Oral contraception with both preparations induced a similar, significant rise in both ceruloplasmin and pregnancy-associated protein. Sex hormone binding globulin levels rose significantly with the ethinyl estradiol-desogestrel, but not with the ethinyl estradiol-levonorgestrel combination. Transcortin increased with both preparations, more with the ethinyl estradiol-desogestrel combination. PMID- 2967234 TI - Absence of immunoreactive luteinizing hormone following gonadotropin-releasing hormone agonist therapy in women with endometriosis. AB - Pituitary desensitization following infusion of gonadotropin-releasing hormone (GnRH) is measurable if bioactivity instead of immunoreactivity is considered. We hypothesized that GnRH agonist therapy induces the same kind of desensitization, but that radioimmunoassays (RIA) for gonadotropins based on polyclonal antibodies cannot show this effect because they recognize inactive fragments of gondadotropins. To test this hypothesis we measured luteinizing hormone (LH) with two different assays: one RIA was based on a polyclonal rabbit anti-hLH, while the other one was an immunoradiometric assay (IRMA) based on 2 different mouse monoclonal anti-hLH. LH measurements were performed on plasma samples obtained from 13 women with laparoscopically proven endometriosis and treated with microcapsules of the GnRH agonist D-Trp6-GnRH (Ferring) once a month. The correlation between LH measurements with both assays in 36 control plasma samples and in another 13 samples obtained before treatment in women with endometriosis was excellent (r = 0.959). In contrast, in women treated with GnRH agonist, the RIA yielded values ranging from undetectable to 12 mIU/ml, whereas 60 out of 66 values were undetectable with the IRMA. We conclude that the monoclonal anti-hLH antibodies in the IRMA either recognize an epitope close to the active site and/or do not recognize the biologically inactive LH fragments which are known to be produced during GnRH agonist therapy. Thus, monoclonal-antibody-based IRMA provide a new and interesting clinical tool to follow the effects of therapies which desensitize the gonadotropic function of the pituitary. PMID- 2967235 TI - [Usefulness of paraspinal muscle electromyography in evaluating low-back pain]. PMID- 2967236 TI - [Late results of the surgical treatment of aortic isthmus stenosis by patch plasty]. AB - To assess long-term outcome after repair of coarctation of the aorta by the patch graft technique of Vossschulte, analysis of the follow-up studies of 66 patients was carried out. At the time of operation, all patients were older than one year of age, mean age 14.4 years (range 1 1/2 to 58 years). The follow-up examinations were performed at a mean of nine years (range five to 17 years) after surgery. Angiographic studies were carried out in all patients with hypertension (ten cases) as well as in those who had undergone surgery ten or more years previously (19 cases). Hypertension was attributed to recurrence of the stenosis in two patients, renal factors were considered responsible in three and in five patients the hypertension was regarded as essential. No patient showed evidence of aneurysm at the site of repair. The fact that this finding is in contrast with the results of other investigators seems most likely related to the surgical technique employed in which the integrity of the intimal membrane is vigently respected to avoid changes in the structure of the aortic wall. PMID- 2967237 TI - Effect of supplementary vaccination in healthy non-responders to hepatitis B vaccination. AB - Among 831 healthy hospital employees vaccinated in accordance with a standard hepatitis B vaccination schedule (three doses of 20 micrograms i.m.), we found 38 (4.6%) hypo- and non-responders with an anti-HBs level less than 10 IU/l. Additional vaccinations with up to three standard doses of hepatitis B vaccine were given to 26 individuals, nine of whom developed anti-HBS titers higher than 10 IU/l after one additional vaccination. Two subjects received no further vaccination. The remaining group of 15 persons were given 2 more injections. After six vaccinations, nine subjects had an anti-HBs response of more than 10 IU/l, while six still had no detectable anti-HBs. In conclusion, supplementary vaccination of healthy hypo- and non-responders after standard hepatitis B vaccination induced an anti-HBs titer greater than 10 IU/l in 38% after one and in 75% after three additional doses of 20 micrograms of hepatitis B vaccine given intramuscularly. PMID- 2967238 TI - Verapamil induces protection of alloxan but not streptozotocin-induced diabetes in rats. PMID- 2967240 TI - Dynamic exercise stimulates ANF secretion by mechanisms independent of prostaglandins. PMID- 2967239 TI - Exercise induced increase in plasma atrial natriuretic peptide and effect of sodium loading in normal man. AB - Atrial tachyarrhythmias and atrial pacing are associated with increased cardiac secretion of atrial natriuretic peptide (ANP) in man. Using treadmill exercise to exhaustion, we have studied the effect of exercise induced tachycardia on plasma immunoreactive ANP (IR-ANP) and vasoactive hormones in 6 normal men before and after 6 days of sodium loading (salt supplements and 0.4 mg 9 alpha fludro hydrocortisone daily for 4 days). Similar increases in heart rate and plasma catecholamine levels occurred during exercise in both studies. Sodium loading increased resting supine plasma IR-ANP (P less than 0.037) and suppressed plasma renin and aldosterone, including the renin-aldosterone response to exercise. Plasma IR-ANP increased more than 3-fold during exercise to 48 +/- 7 before and 66 +/- 12 pmol/l after sodium loading (P greater than 0.1). When the response of individual subjects was examined, there was no significant correlation between change in plasma IR-ANP and change in heart rate or catecholamine levels in either exercise study. Exercise induces greater increments in plasma IR-ANP than either acute or chronic sodium loading in normal men and may be a useful and rapid means of assessing the heart's ability to secrete ANP. PMID- 2967241 TI - Physiological changes in androgen plasma levels with elapsing of time from castration in adult male rats. AB - In the present study we investigated in adult male rats the effects of castration on Dehydroepiandrosterone (DHEA), Androstenedione (delta 4), Testosterone (T) and Dihydrotestosterone (DHT) plasma levels: five days (group II), seven weeks (group III) and eleven weeks (group IV) after orchiectomy. The same hormone assays were performed in rats approximately 60 days of age which underwent a sham-operation for orchiectomy (group I). Our data show that five days following orchiectomy (group II) delta 4, T and DHT were decreased with respect to sham-operated rats. (Group I: delta 4: 83.3 +/- 14.9 (SEM) ng/dl (n = 12); T: 435.32 +/- 51.45 (n = 12); DHT: 51.47 +/- 6.54 (n = 12); Group II: delta 4: 44.81 +/- 6.09 (n = 12) P = 0.05; T: 25.54 +/- 2.88 (n = 12) P less than 0.01; DHT: 12.9 +/- 2.51 (n = 12) P less than 0.01). Seven weeks afterwards T and DHT remained significantly lower (group III: T: 54.37 +/- 12.21, n = 16) (P less than 0.01; DHT: 33.22 +/- 4.49 (n = 16) P less than 0.01) while eleven weeks after all steroids were significantly decreased with respect to the values observed in sham-operated rats. (Group IV) delta 4: 32.01 +/- 5.7 (n = 10) P less than 0.01: T: 27.29 +/- 7.05 (n = 10) P less than 0.01; DHT: 29.03: 5.34 (n = 10) P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967242 TI - Atrial natriuretic factor in cirrhosis with ascites: plasma levels, cardiac release and splanchnic extraction. AB - The aim of this study was to determine the plasma levels, cardiac release and splanchnic extraction of atrial natriuretic factor in cirrhosis with ascites. The plasma concentration of immunoreactive atrial natriuretic factor in samples obtained from an antecubital vein was measured in 18 healthy volunteers and in 35 cirrhotics with ascites. In 11 of these cirrhotics and in 11 patients admitted to the hospital for the study of a thoracic pain who had no clinical or hemodynamic signs of cardiac failure (control group), the plasma levels of immunoreactive atrial natriuretic factor in samples from the coronary sinus, right atrium, pulmonary artery, hepatic vein and femoral vein were determined and the coronary sinus blood flow measured by thermodilution. Cirrhotic patients showed significantly higher plasma levels of immunoreactive atrial natriuretic factor in each vascular territory studied than did control subjects (coronary sinus: 101.2 +/- 10.6 vs. 26.1 +/- 4.7 fmoles per ml; right atrium: 32.5 +/- 5.8 vs. 9.4 +/- 3.5; pulmonary artery: 36.8 +/- 10.1 vs. 7.5 +/- 2.4; hepatic vein: 10.7 +/- 2.0 vs. 2.7 +/- 0.8; femoral vein: 18.2 +/- 2.4 vs. 3.1 +/- 0.9; antecubital vein: 14.7 +/- 1.6 vs. 4.0 +/- 0.8). The coronary sinus blood flow was also higher in cirrhotics (200 + 22 ml per min) than in controls (105 +/- 7 ml per min). Consequently, the estimated cardiac release and cardiac production of immunoreactive atrial natriuretic factor were strikingly increased in cirrhotics (13,334 +/- 2,007 and 5,484 +/- 1,734 fmoles per min, respectively) as compared to control subjects (1,669 +/- 338 and 1,431 +/- 350 fmoles per min, respectively; p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967243 TI - A human complement-fixing monoclonal anti-human lymphocyte antibody of rat origin. AB - MARCH 1E11 is an IgM monoclonal anti-human lymphocyte antibody of rat origin with the capacity to utilise both human and rabbit complement. The antibody reacts with all thymocytes and with all peripheral blood T and B lymphocytes. The treatment of human bone marrow or human peripheral blood mononuclear cells (PBMC) with MARCH 1E11 and either pooled human serum or autologous serum as a complement source resulted in cytolysis of greater than 99% of OKT3-positive lymphocytes. Under these conditions, progenitor cell recovery (colony-forming unit (CFU-c), burst forming unit--erythroid (BFU-e) and colony-forming unit--mixed (CFU-mix)) was greater than 90% of that of untreated cells. The response of treated marrow or PBMC to phytohaemagglutinin stimulation and in mixed leucocyte reactions demonstrated a reduction in thymidine incorporation to values similar to those obtained for unstimulated cells. The antibody does not cause modulation of the cell surface antigen and does not react significantly with non-lymphoid tissues. This monoclonal antibody may be useful for in vitro elimination of T lymphocytes from allogeneic bone marrow used for transplantation. The antibody may also be useful for treatment protocols requiring lymphoid depletion or immunosuppression as in organ transplantation. PMID- 2967244 TI - Functional deficiencies of spleen dendritic cells in autoimmune MRL/lpr mice. AB - We investigated functional aspects of splenic dendritic cells (DC) obtained from MRL/MpJ-lpr/lpr (MRL/l) mice, and their normal counterparts, MRL/MpJ-+/+ (MRL/+) mice. In vitro antigen-presenting activity of DC obtained from MRL/l mice was impaired compared with that of DC from MRL/+ mice. Another major functional aspect of DC, i.e., stimulator cell activity in the allogeneic mixed lymphocyte reaction (MLR), was also found to be impaired in MRL/l mice. MRL/l mice and MRL/+ mice were also examined for co-operation between macrophages (M luminal diameter) and DC. We found peritoneal resident M luminal diameter obtained not only from MRL/+ mice but also from MRL/l mice released a soluble factor which enhanced the activity of DC from the respective mice. Furthermore, DC from both MRL/l and MRL/+ mice responded to the culture supernatant of a mouse macrophage hybridoma clone, S44, which releases a factor that enhances the antigen-presenting activity of DC, and their activity as stimulator cells in the allogeneic MLR was enhanced. However, the degree of enhancement was less in MRL/l mice than in MRL/+ mice. PMID- 2967245 TI - The effect of dietary restriction on mouse T cell functions. AB - Forty percent dietary restriction on 9-weeks-old C3H/He mice caused decrease of the weight of central lymphoid organs in parallel with the reduction of body weight. However, the percentage of splenic T cells was dramatically increased in diet-restricted mouse spleen cells. Generally, normal mouse spleen cells contained about 30% of Thy 1.2+ T cells, but the restricted mouse spleen cells contained 80% Thy 1.2+ T cells. Ly 1+, L3T4+ T cells, but not Ly 2+ T cells, also increased in diet-restricted mouse compared with the unrestricted mice. In parallel with the dramatic changes of splenic T cells, spleen cells obtained from diet-restricted mice showed higher immunological responses against alloantigen and interleukin 2. It was also demonstrated than nylon-passed splenic T cells obtained from diet-restricted mice showed higher levels of T cell responses against r-IL-2 and alloantigen, indicating that dietary restriction modulates T cell functions themselves. PMID- 2967246 TI - Antimicrobial sensitivity of Neisseria gonorrhoeae to spectinomycin & rosoxacin. PMID- 2967247 TI - Monoclonal antibodies to Staphylococcus aureus laminin-binding proteins cross react with mammalian cells. AB - We and others have previously shown that some microorganisms, including bacteria, express on their surfaces receptors that specifically recognize extracellular matrix proteins, such as laminin, fibronectin, or both. The ability of microorganisms to adhere and to invade might depend on the existence of receptors which could, thus, be correlated with pathogenicity. In the present paper, we report the isolation of five stable cell lines that were producers of monoclonal antibodies to Staphylococcus aureus laminin receptors. One of these antibodies, which was of the immunoglobulin M isotype, blocked the binding of laminin to bacteria before and after fixation and recognized the putative 52-kilodalton laminin-binding protein in whole bacterial extracts. Also, purified receptor was isolated by immunoaffinity chromatography and shown to bind laminin. Furthermore, the same antibodies bound the 67-kilodalton putative receptor from mouse melanoma cells and gave positive immunofluorescence reactions against mammalian tumor cells. These data strongly suggest either the evolutionary conservation of at least some sequences in both procaryotic and eucaryotic laminin-binding proteins or convergent evolution and positive selection of epitopes cross-reacting with laminin. Some of these antibodies to the procaryotic protein could therefore become useful markers for the expression of laminin receptors by cancer cells. PMID- 2967249 TI - Hepatitis B in a nonresponder to hepatitis B vaccine. PMID- 2967250 TI - Mast cell degranulation as a major event in the effector phase of delayed-type hypersensitivity induced by cloned helper T cells. AB - A kinetic study (0-72 h) was performed on the cellular composition of the exudate in delayed-type hypersensitivity (DTH) induced by the injection of cloned helper T cells into the footpad of C57BL/6J mice. In this model the reaction was maximal at 24 h, as assessed by local swelling. After an initial phase of inflammatory cell infiltration, we observed pronounced degranulation of mast cells, occurring at 15-19 h. Administration of antihistamines shortly before this time significantly inhibited the increase in footpad thickness at 24 h, suggesting mast cell mediators as a major cause of swelling in DTH. Furthermore, a new arrival of inflammatory cells, particularly eosinophils, was clearly correlated with mast cell degranulation. The latter, which might function to suppress local reactions, persisted as the only intact cell type after 48 h. Supplementary examination of peripheral blood smears showed successive peaks of neutrophils (at 3 h), monocytes (at 15 h) and finally eosinophils (after 20 h). These results suggest a local and systemic cascade in DTH which depends on mast cell degranulation for its full expression. PMID- 2967248 TI - Sequence analysis of the glucosyltransferase A gene (gtfA) from Streptococcus mutans Ingbritt. AB - The complete nucleotide sequence of a 2.4-kilobase fragment containing the glucosyltransferase A gene (gtfA) of Streptococcus mutans Ingbritt (serotype c) was determined. The gtfA gene contains 481 codons and specifies a protein of molecular weight 55,665. There is no evidence of a signal peptide in the protein or that the glucosyltransferase A enzyme is secreted. No sequence homologies were observed between the gtfA gene or protein and the gtfI or gtfB gene and its protein. PMID- 2967251 TI - Syncope as the presentation of unstable angina. AB - A 71-year-old woman presented with syncope in association with unstable angina. Coronary angiography and subsequent postmortem findings demonstrated severe coronary arterial disease. The importance of the association of syncope with unstable angina and possible underlying mechanisms are discussed. PMID- 2967252 TI - The combined use of diastolic counterpulsation and coronary dilation in unstable angina due to multivessel disease under unstable hemodynamic conditions. AB - Sixteen patients with multivessel ischemic heart disease and severely jeopardized myocardium required intra-aortic balloon counterpulsation subsequent to a deterioration in hemodynamics during or following a coronary angioplasty procedure. They had all suffered unstable angina which was refractory to intensive medical therapy, consisting of a combination of nitroglycerin, beta adrenergic antagonists, and calcium blockers. Thirty angioplasties had been attempted (1.9 artery stem/patient) with a primary success rate of 90%. The symptoms of prolonged myocardial ischemia had disappeared, and the patient's blood pressure had normalized. No complications were associated with the use of the mechanical circulatory assistance. There were no deaths related to the procedure itself, and no myocardial infarctions. Emergency surgery was not required. One patient did die in hospital, however, due to cerebrovascular accident which occurred 4 days after removal of the mechanical circulatory support. Two also died suddenly later. One patient also required later elective coronary arterial bypass surgery and another needed repeated coronary dilation. The 12 remaining patients are asymptomatic at a follow-up with mean value of 22 months. Temporary intra-aortic diastolic counterpulsation is a useful adjunct to coronary angioplasty in patients with multivessel unstable angina and compromised hemodynamics. PMID- 2967253 TI - Transluminal balloon dilation of resected coarcted segments of thoracic aorta: histological study and clinical implications. AB - The effect of experimental balloon dilation on seven excised specimens of classic coarctation was assessed histologically. Widening of the lumen was achieved in all specimens by tears in the vessel wall. The tears extended through the intima in one specimen and through both intimal and medial layers in six specimens. In view of the extent of wall damage, we postulate that there would be serious long term complications following balloon dilation for the treatment of native coarctation. PMID- 2967254 TI - The nail. PMID- 2967255 TI - [Postoperative hemorrhagic complication in a patient with type I von Willebrand disease following surgical extraction of an impacted tooth after preparation with DDAVP: a case report]. PMID- 2967256 TI - Changes in skin perfusion after sympathetic block with guanethidine. Laser Doppler flowmetry in human volunteers. AB - Total forearm blood flow and skin microcirculation have been measured by occlusion plethysmography and by laser Doppler flowmetry (skin blood cell flux) before and after the induction of regional sympathetic block with Guanethidine. Total forearm blood flow more than doubled while the skin blood cell flux increased by 50% comparing simultaneous flows of blocked and control arms. The increase lasted for 3 days during which period the flux pattern registered by the laser Doppler flowmeter in the blocked extremities showed regular variations oscillating around the mean flux in an almost sinus wave fashion. In non-blocked arms the flux pattern registered by the laser Doppler flowmeter was highly irregular. We conclude that regional sympathetic block with Guanethidine results in an increased skin microcirculation in healthy human volunteers and that the increase lasts for 3 days. No valid conclusions can be made from this study concerning the nutritive benefit of the microcirculatory change. PMID- 2967257 TI - Constant-pressure perfusion on the isolated rat liver: local oxygen supply and metabolic function. AB - Isolated rat livers were perfused with a haemoglobin-free Krebs-Ringer solution containing 35 g albumin/1. During a 3 hour perfusion total flow, O2-uptake rate and local O2-supply remained constant. During the same period of time the ATP content increased from a value of 0.59 +/- 0.17 mumol/g to 1.81 +/- 0.42 mumol/g. The low initial value was due to the ischaemic period during preparation of the organ. The glycogen content increased by 13.4 mg/g, indicating that the organ did not lose the ability for glyconeogenesis. In addition to an increase in glycogen, there was a continuous increase in the glucose concentration of the perfusate, while lactate was taken up by the organ. The results indicate that the isolated organ can be perfused for at least 3 hours without development of disturbances in local O2-supply. PMID- 2967258 TI - Plasma fibrinopeptide A, beta-thromboglobulin, and platelet factor 4 in diabetic retinopathy. AB - We examined plasma levels of fibrinopeptide A, beta-thromboglobulin, and platelet factor 4 in diabetic patients. Among diabetic patients (n = 33) plasma levels of fibrinopeptide A, beta-thromboglobulin and platelet factor 4 were significantly higher than in controls (n = 41). In the subgroups of diabetic patients with (1) minimal (n = 13), and (2) moderate-severe (n = 14) retinopathy only plasma fibrinopeptide A levels were significantly higher than in controls. Among the total group of diabetic patients plasma levels of fibrinopeptide A increased significantly with increasing severity of retinopathy. These results suggest that diabetic retinopathy is associated with in vivo activation of blood coagulation factors and that this activation increases with advancing retinopathy. PMID- 2967259 TI - Corneal endothelial morphology in the rat. Effects of aging, diabetes, and topical aldose reductase inhibitor treatment. AB - Increases in corneal endothelial cell polymegathism and pleomorphism are characteristic of diabetic human and dog corneas. This study investigated the rat as a model for age- and diabetes-related changes in endothelial cell morphology. As in most mammals, aging of the normal rat results in a progressive decrease in cell density as well as reduced numbers of hexagonal endothelial cells and increased coefficient of variation of cell size after age 34 weeks. Streptozotocin-induced diabetes produced an early progressive increase in the coefficient of variation of cell size and decrease in percentage of hexagonal cells so that diabetic rats were significantly different from age-matched normal rats by 24 weeks of age. Topical treatment with the aldose reductase inhibitor, AL 1576, begun immediately after diabetes induction, prevents endothelial cell changes and cataract formation. Topical aldose reductase inhibition also reverses endothelial cell changes when treatment is begun 8 weeks after streptozotocin injection. These results indicate that the rat is a good model for studying diabetes-induced corneal endothelial changes and that topical aldose reductase inhibitors may be effective in preventing or reversing diabetic corneal endothelial cell changes. PMID- 2967260 TI - Diminished cerebrospinal fluid beta-endorphin concentration in monkeys with arachnoiditis. AB - Altered concentrations of brain beta endorphin and spinal cord encephalin have been reported in mice with iatrogenic arachnoiditis. We measured beta-endorphin concentration in cerebrospinal fluid (CSF) in normal monkeys and in monkeys with various degrees of arachnoiditis resulting from myelography and spinal surgery and found a significant negative correlation between the arachnoiditis score and the lumbar CSF beta-endorphin concentration. This study suggests that an assay of CSF beta endorphin can be used to detect the alterations in neurotransmitters in arachnoiditis. PMID- 2967261 TI - Effects of angioplasty balloon inflation time on arterial contractions and mechanics. AB - The purpose of this study was to characterize and compare the immediate effects of short and long angioplasty inflation times on arterial contractions and passive mechanics and thereby determine if there is a potential advantage obtained by increasing the duration of balloon inflation. In each of 10 nonatherosclerotic New Zealand rabbits, one external iliac artery was dilated for 20 seconds, and the contralateral artery was dilated for 2 minutes. Although angioplasty stretched the arteries 27% and 30% for the short and long dilations respectively, the pre- and post-angioplasty arteriographic diameters were not different. Both short and long dilations had equal effects on passive biomechanics: circumferential wall stress was increased (P less than .01); wall thickness was decreased (P less than .01); the incremental elastic modulus was increased (P less than .01). In vitro studies of arterial rings demonstrated that maximal active contractile force in response to KCl (70 mM) was significantly (P less than .05) less for dilated arteries than for undilated arteries. More importantly, maximal active force after the 2-minute dilations was significantly (P less than .05) less than after the 20-second dilations. These results suggest that, when dilating normal arteries, increasing angioplasty balloon inflation time from 20 seconds to 2 minutes offers no mechanical advantage but produces more smooth muscle cell dysfunction, which may reduce vasospasm and restenosis after angioplasty. PMID- 2967262 TI - Sanctioned assisted suicide: "separate but equal" treatment for the "new illegitimates". PMID- 2967263 TI - The salvage of the lower limbs: choice of the prosthetic material in femoro distal and axillo-femoral bypass. AB - The choice of the prosthetic material in arterial surgery of patients with rest pain and/or impending gangrene of the lower limbs still presents several problems. The poor run-off, the small caliber of the distal vessels, the crossing of the joint areas and the length of the bypass may lead to early occlusion of the prosthesis. This experience is based on 268 femoro-distal (214 femoro popliteal below the knee and 54 femoro-tibial) and on 121 axillo femoral/popliteal bypasses (89 axillo-femoral, 28 axillo-bifemoral and 4 axillo popliteal). In the 389 surgical procedures we have employed the autologous saphenous vein in 208 cases, Polytetrafluoroethylene (PTFE) straight or tapered in 66, PTFE external supported (EXS) in 33, Dacron in 12, homologous saphenous vein in 7 and PTFE EXS Thin Wall in 5. In 58 cases a composite graft (autologous saphenous vein plus synthetic prosthesis) was used. The cumulative long term (12 96 months) patency is 75.96% for autologous saphenous vein bypass, 62.12% for PTFE, 75.75% for PTFE EXS, 41.66% for Dacron, 42.85% for homologous saphenous vein and 84.48% for the composite graft. All the PTFE EXS Thin Wall grafts are still patent (12 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967264 TI - Short- and long-term results of angioplasty for multiple coronary stenoses. AB - Sixty-eight patients underwent percutaneous transluminal coronary angioplasty (PTCA) for multiple coronary stenoses. Lesions involved one coronary vessel in 24 patients, two in 34, and three or more vessels in 10 patients. The primary patient success rate was 96%, and we successfully dilated 162 (91%) of 179 significant (i.e., greater than 70% obstructive) coronary lesions, mean 2.4 lesions/patient. Complete coronary revascularization was achieved in 24 patients (Group 1), while major narrowings were not dilated either by intention or due to failure of dilation in 42 others (Group 2). The clinical state of the patients improved markedly after successful PTCA, and 44 (72%) of 61 were in the New York Heart Association Functional Class I or II 3 to 24 months later. In keeping with the clinical results, there was a significant improvement in treadmill exercise performance, in both groups. Patients with complete revascularization, however, tended to exercise longer (NS) to a greater heart rate (P less than 0.004) with less or no ischemic ST depression on ECG (P less than 0.04). Complications were few and limited to patients undergoing PTCA for unstable angina pectoris or following acute myocardial infarction. One patient (1%) died, two (3%) had Q-wave infarction and one patient sustained a cerebral embolus during catheterization. Restenosis was observed in 6 (29%) of 21 consecutive patients recatheterized 6 to 12 months (or sooner for symptoms) after successful PTCA. Four patients underwent a repeat PTCA, one was referred for coronary artery bypass grafting (CABG), and in one restenosis was minor and symptoms were mild. PTCA should be considered in patients with multiple coronary stenoses in whom all or most of the significant lesions can be dilated. Partial revascularization by PTCA is clinically useful and may be preferable to CABG in patients in whom the surgical risk is increased. PMID- 2967265 TI - Melphalan attenuates streptozotocin-induced diabetes in C57BL/6 and C3H3 mice. PMID- 2967266 TI - The effects of immunosuppressive pharmacological agents on the induction of cytotoxic and suppressor T lymphocytes in vitro. AB - The immune system is regulated by the interactions among several distinct functional subsets of T cells. The action of several commonly used immunosuppressive drugs on the activation of cytotoxic T lymphocytes (CTL) and suppressor T lymphocytes (Ts) in the primary mixed lymphocyte reaction (MLR) was investigated. Cyclosporin A, hydrocortisone, and azathioprine were all found to inhibit both CTL and Ts activation when present at pharmacological doses in culture. When these drug-inhibited cultures were reconstituted with interleukin 2, however, clear differences between the effects of these drugs was observed. Cyclosporin A and hydrocortisone allowed the selective activation of Ts in the presence of interleukin-2, while azathioprine inhibition was not reversed by interleukin-2. Thus, CTL precursors appear to be directly inhibited by all of these drugs, but Ts precursors apparently are not inhibited by cyclosporin A or hydrocortisone provided interleukin-2 is present. These findings are discussed in terms of the activation requirements of CTL vs. Ts and the implications of the selective activation of alloantigen-specific Ts for prevention of allograft rejection. PMID- 2967267 TI - Enhancement of human T lymphocyte functions by prothymosin alpha. I. Augmentation of mixed lymphocyte culture reactions and soluble protein-induced proliferative responses. AB - Prothymosin alpha (ProT alpha), a 115-amino-acid thymic polypeptide, was tested for its effect on soluble antigen, allo- and auto-antigen-induced human T-cell proliferation. ProT alpha enhanced the secondary T-cell proliferative response to ovalbumin (OVA)- and keyhole limpet haemocyanin (KLH)-pulsed antigen-presenting cells (peripheral blood monocytes). Maximum enhancement (20-fold for OVA and 23 fold for KLH) occurred when suboptimal concentrations of either OVA or KLH were employed. Subset depletion experiments showed that the helper/inducer T-cell subpopulation was responsible for the observed enhancement. In the mixed lymphocyte reaction (MLR), ProT alpha enhanced autoantigen- (autoMLR; 9- to 14 fold) as well as the alloantigen- (alloMLR; 8- to 10-fold) induced T-cell proliferation when suboptimal ratios of the participating cells were used. Preincubation of the stimulating (autologous or allogeneic monocytes) with ProT alpha induced significantly higher T-cell proliferation in both primary and secondary MLR responses as compared to that induced by non-treated monocytes. In contrast, T lymphocytes pre-incubated with ProT alpha did not show enhanced proliferative activity when tested subsequently in the MLR. Suboptimal numbers of T cells exhibited high proliferative activity when pre-incubated with ProT alpha in the presence of autologous monocytes. These studies suggest that ProT alpha potentiates T-cell proliferative responses not directly, but via monocytes which are included in the cultures either as antigen-presenting cells or accessory and/or stimulator cells. The importance of ProT alpha in pathologically occurring defective cellular immune response systems discussed. PMID- 2967268 TI - [Oral surgery in patients in the "3d age" with systemic diseases]. PMID- 2967269 TI - Ultrastructural localization of steroid sulphatase in cultured human fibroblasts by immunocytochemistry: a comparative study with lysosomal enzymes and the mannose 6-phosphate receptor. AB - Immunocytochemistry was used to study the subcellular localization of steroid sulphatase in cultured human fibroblasts. Ultra-thin cryosections were incubated with antibodies raised against steroid sulphatase purified from human placenta and immune complexes were visualized with gold probes as electron dense markers. Steroid sulphatase was found in rough endoplasmic reticulum, Golgi cisternae and in the trans-Golgi reticulum, where it co-distributes with lysosomal enzymes and the mannose 6-phosphate receptor. The enzyme was not detected in lysosomes. Steroid sulphatase was also found at the plasma membrane and in the endocytic pathway (i.e. coated pits, endosomes and multivesicular endosomes). These may be the sites where sulphated oestrogen precursors are hydrolysed. Also here, it co localizes with lysosomal enzymes and the mannose 6-phosphate receptor. It is concluded that microsomal steroid sulphatase and lysosomal enzymes share several cellular compartments. PMID- 2967271 TI - [Depo-Provera--antineoplastic agents which is also a contraceptive]. PMID- 2967270 TI - Histochemical heterogeneity of intrafusal muscle fibres in slow and fast skeletal muscles of the rat. AB - Intrafusal muscle fibres of the slow soleus (Sol) and fast vastus lateralis (VL) muscles of the rat were studied histochemically. Serial transverse sections were incubated for the localization of succinate dehydrogenase (SDH), alpha glycerophosphate dehydrogenase (GPD) and adenosine triphosphatase (ATPase). The latter was examined further after preincubation in acidic solution held at either low or room temperature (RT). The bag2 intrafusal fibres in both muscles displayed high regular and acid stable ATPase, but low SHD and GPD activities. Bag1 intrafusal fibres showed low to moderate regular ATPase, a regional heterogeneity after RT acid preincubation (low activity in juxtaequatorial and high in polar zones), moderate SDH, but low GPD reactions. In both muscles the chain fibres usually exhibited high ATPase for both regular and cold acid preincubated reactions, but usually low activity after RT acid preincubation; they had high SDH but variable GPD activities. In Sol muscle, however, approximately 25% of spindles contained chain fibres that showed high acid-stable ATPase reaction after both cold and RT acid preincubation. In contrast, chain fibres in some VL spindles had a characteristically low ATPase reaction even after cold acid preincubation. This study, therefore, has delineated the existence of an inherent heterogeneity among chain fibres (with respect to their histochemical reactions) in muscle spindles located within slow and fast muscles and also between those found within populations of either Sol or VL muscle spindles. PMID- 2967272 TI - An antitumor polypeptide antibiotic neocarzinostatin: the mode of apo-protein- chromophore interaction. AB - The mode of neocarzinostatin-chromophore (NCS-chr)-apo-neocarzinostatin (apo-NCS) interaction in neocarzinostatin (NCS) complex has been described. The NCS-chr release from the NCS complex in the presence of various reagents, which destroy the high-order structure of the protein under various pH conditions, was examined. We found that (i) sodium dodecylsulfate, Nonidet P 40, 8 M urea, and 2 propanol did release NCS-chr from NCS, (ii) no NCS-chr release is detected below pH 7, but it is enhanced at high pH and (iii) beta-naphthol as a model of naphthalenecarboxylic acid derivative and D-galactosamine as a model of N methylfucosamine of NCS-chr did release NCS-chr from NCS. These observations indicate that the binding of NCS-chr to apo-NCS may be due to not only ionic interaction between the acidic side chain of apo-NCS and the basic center of an aminosugar moiety of NCS-chr but also hydrophobic interaction between the hydrophobic amino acids of apo-NCS and hydrophobic moieties of NCS-chr. Apo-NCS is a very hydrophilic protein, since it has an high hydrophilic amino acid content. So, local hydrophobicity, local hydrophilicity and secondary structure of apo-NCS were predicted. Hydrophobic residues of apo-NCS predominantly located in beta-sheet structures near the carboxyl-terminus. These predictions are in good agreement with the results suggesting that NCS-chr bound carboxyl-terminal 43-peptide of apo-NCS in our previous result. PMID- 2967273 TI - Effect of zearalenone on days 7 to 10 postmating on intrauterine environment and migration of embryos in sows. AB - On d 7 to 10 postmating, first-litter sows were fed either a control diet or a diet containing zearalenone (ZEN; 1 mg/kg body weight). Surgery was performed on either d 9, 11 or 13 postmating to collect blastocysts and uterine flushings. The rostral and caudal portion of each uterine horn was flushed with phosphate buffered saline, and the blastocysts were separated from the recovered solution. Uterine flushings were analyzed for total Ca, Mg, Zn, estradiol-17 beta (E2 17 beta) and progesterone (P4). Administration of ZEN did not affect the number of blastocysts recovered or the position of embryos within the uterus on d 9 or 11. Blastocysts recovered on d 13 were filamentous and could not be enumerated. Total Ca in uterine flushings of control sows was higher (P less than .001) on d 11 than on d 9 or 13, but intrauterine Ca of ZEN-treated sows did not vary by sampling day (P greater than .05) and was lower (P = .01) than that of controls on d 11. Total intrauterine Mg of ZEN sows was greater (P = .002) than of control sows on d 11 and 13, and total intrauterine Zn of ZEN sows was greater than that in control sows on d 13. There were no differences in total intrauterine P4 or E2 17 beta among ZEN-treated and control sows on d 9, 11 or 13 postmating. Serum concentrations of 13, 14-dihydro-15-ketoprostaglandin F2 alpha (PGFM) increased from d 9 to 13 in control and ZEN-treated sows, but there were no differences between treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967274 TI - Identifying reinforcers for persons with profound handicaps: staff opinion versus systematic assessment of preferences. AB - We evaluated a systematic means of determining stimulus preferences among seven profoundly handicapped persons. Preferences were determined by observing student approach responses to individual stimuli. Results indicated that there were differential stimulus preferences across the multiply handicapped participants. However, results of the systematic assessment did not coincide with the results of a more traditional, caregiver-opinion method of assessing student preferences. A second experiment was then conducted with five participants to evaluate whether stimuli that were assessed to consistently represent preferences would function as reinforcers in skill training programs. Results indicated that stimuli that were systematically assessed to represent student preferences typically functioned as reinforcers when applied contingently. However, preferred stimuli as reflected by caregiver opinion did not function as reinforcers unless those stimuli were also preferred on the systematic assessment. Results are discussed in terms of assisting profoundly handicapped persons by (a) improving the effectiveness of training programs by increasing the likelihood of using stimuli that have reinforcing value and (b) increasing the overall quality of life by providing preferred stimuli in the routine living environment. PMID- 2967276 TI - Rib cage and abdominal expiratory muscle responses to CO2 and esophageal distension. AB - The present study compared the responses of rib cage and abdominal expiratory muscles to chemical and mechanical stimuli. In pentobarbital-anesthetized spontaneously breathing dogs, electromyograms (EMG) were recorded from the triangularis sterni (TS) and transverse abdominis (TA) muscles using bipolar intramuscular wire electrodes. During resting oxygen breathing, both muscles were electrically active during expiration. Progressive hyperoxic hypercapnia significantly augmented the expiratory activity of both the TA and the TS. However, the mean percent increases in electrical activity in response to CO2 were substantially greater for the TA than for the TS at all PCO2 levels greater than 50 Torr (P less than 0.01). Occlusion of the airway at end inspiration significantly delayed the onset of TS EMG (from 0.35 +/- 0.07 to 3.35 +/- 0.67 sec; P less than 0.002) and decreased TS EMG rate of rise (P less than 0.002), but did not significantly alter these parameters for the TA. Esophageal distension increased TS EMG in all dogs (by mean of 220 +/- 64%; P less than 0.01), but in contrast decreased TA EMG in all dogs (by a mean of 63 +/- 12%; P less than 0.001). The response to esophageal distention occurred in a graded manner and appeared to be mediated predominantly via vagal afferents. We concluded that expiratory muscles of the rib cage and abdomen manifest substantial differences in their electrical responses to chemoreceptor, pulmonary stretch receptor, and esophageal mechanoreceptor stimuli.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967275 TI - Transverse abdominis length changes during eupnea, hypercapnia, and airway occlusion. AB - The abdominal muscles accelerate airflow during expiration and may also influence the end-expiratory volume and configuration of the thorax. Although much is known about their electrical activity, the degree to which they change length during the respiratory cycle has not been previously assessed. In the present study we measured respiratory changes in transverse abdominis length using sonomicrometry in 14 pentobarbital sodium-anesthetized supine dogs and compared length changes to simultaneously recorded tidal volume and transverse abdominis electromyograms (EMG). To determine muscle resting length at passive functional residual capacity (LFRC), the animals were hyperventilated to apnea. The transverse abdominis was electrically active in all animals during resting O2 breathing (eupnea). During inspiration the transverse abdominis lengthened above resting length in all 14 dogs by a mean of 3.7 +/- 1.1% LFRC; during expiration the transverse abdominis shortened below resting length in 13 of 14 dogs by a mean of 4.2 +/- 0.9% LFRC. Increasing hyperoxic hypercapnia (produced in 9 animals) progressively heightened transverse abdominis EMG and progressively increased the extent of muscle shortening below resting length (to 12.6 +/- 3.2% LFRC at a PCO2 of 90 Torr). During single-breath airway occlusion substantial inspiratory lengthening of the transverse abdominis occurred, both during O2 breathing and during CO2 rebreathing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967278 TI - Healthy mothers, healthy babies. PMID- 2967277 TI - Effects of exercise on cardiac myosin isozyme composition during the aging process. AB - The effects of aging and exercise on isoforms of cardiac myosin and Ca2+ activated actomyosin adenosinetriphosphatase (ATPase) activity were examined in Fischer 344 rats. Rats were divided into running (R) and age-matched sedentary (S) groups. The groups initiated their exercise program at either 3, 4, or 18 mo of age. Rats were killed at 10, 12, 24, or 27 mo of age. ATPase activity decreased 25% in the S group and 28% in the R group from 12 to 27 mo of age. The myosin isozyme patterns shifted in both S and R groups from a predominantly V1 isozyme form (63.8%) at 10 mo of age to a more equal distribution of isozyme forms at 24 mo (V1, V2, and V3 comprising 40.0, 27.8, and 31.9%, respectively). Age-related shifts in myosin composition occurred despite chronic endurance training at an intensity of approximately 75% maximum O2 consumption. Improvement of cardiac performance through training during aging is not accompanied by attenuating shifts in myosin isozyme composition. PMID- 2967279 TI - HIV-related malignancy. PMID- 2967280 TI - Interdisciplinary approach to arthritis rehabilitation. PMID- 2967281 TI - Controlling the medical magazine monster (or how not to be buried alive under a mountain of journals). PMID- 2967282 TI - Maintaining professional autonomy: strategies for staying in charge. PMID- 2967283 TI - Interruption of the inferior vena cava above and between the renal veins. PMID- 2967285 TI - The effects of phosphorylation and dephosphorylation of brain myosin on its actin activated Mg2+-ATPase and contractile activities. AB - Purified bovine brain myosin contained approximately 1 and 3 mol of protein-bound phosphate/mol myosin in the light chains and heavy chains, respectively. Large portions of this light chain- and heavy chain-bound phosphate (about 0.8 and 2.4 mol, respectively) were removed by incubation with a brain phosphoprotein phosphatase and potato acid phosphatase, respectively. Upon phosphorylation of the dephosphorylated brain myosin with myosin light chain kinase and casein kinase II, about 1.6 and 3.0 mol of phosphate was incorporated into the light chains and heavy chains, respectively, while much lower levels of phosphate were incorporated into the non-dephosphorylated brain myosin under the same conditions. The actin-activated Mg2+-ATPase activity of brain myosin rephosphorylated with myosin light chain kinase was about twice as high as that of dephosphorylated brain myosin (about 30 and 15 nmol phosphate/mg/min, respectively). On the other hand, whereas the rephosphorylated brain myosin superprecipitated rapidly with F-actin, the rate of superprecipitation of the dephosphorylated brain myosin was extremely low. Under appropriate conditions, a loose network of tiny superprecipitates, which formed initially throughout the solution, contracted to form eventually a large and dense particle. These results indicate that phosphorylation of the light chains of brain myosin is a prerequisite for the contraction of brain actomyosin. The role of phosphorylation of the heavy chains by casein kinase II remains to be elucidated. PMID- 2967286 TI - Comparison of sarcoplasmic reticulum Ca2+-adenosine triphosphatase from vitamin E deficient dystrophic rabbit skeletal muscle with iron-ascorbate-treated and untreated enzyme. AB - Sarcoplasmic reticulum Ca2+-ATPase from rabbit skeletal muscle has an Arrhenius curve of enzyme activity with a discontinuity at about 20 degrees C. Preparations treated with FeSO4 and ascorbic acid and from a vitamin E-deficient dystrophic rabbit have 22% of the normal activity and a linear Arrhenius curve (Promkhatkaew, D., Komaratat, P., & Wilairat, P. (1985) Biochem. Int. 10, 937 943). All three preparations were cross-linked to the same extent by dimethyl suberimidate and copper-phenanthroline reagent at temperatures above and below the temperature of the Arrhenius discontinuity. Both iron-ascorbate-treated Ca2+ ATPase and that from a vitamin E-deficient animal had 50% of the normal sulfhydryl content, but the disulfide and free amino contents were unaltered. These observations suggest that loss of sulfhydryl groups through lipid peroxidation, both in vivo and in vitro, resulted in reduction of Ca2+-ATPase activity and loss of the break in the Arrhenius plot. Changes in Ca2+-ATPase polypeptide aggregational state could not account for the discontinuity in the Arrhenius curve as revealed by the similar extent of cross-linking of the three enzyme preparations at temperatures above and below the temperature of the Arrhenius discontinuity. PMID- 2967284 TI - Coincident localization of secretory and plasma membrane proteins in organelles of the yeast secretory pathway. AB - Immunoelectron microscopy of Saccharomyces cerevisiae cells embedded in Lowicryl K4M has been used to localize invertase and plasma membrane (PM) ATPase in secretory organelles. sec mutant cells incubated at 37 degrees C were prepared for electron microscopy, and thin sections were incubated with polyclonal antibodies, followed by decoration with protein A-gold. Specific labeling of invertase was seen in the lumen of the endoplasmic reticulum, Golgi apparatus, and secretory vesicles in mutant cells that exaggerate these organelles. PM ATPase accumulated within the same organelles. Double-immune labeling revealed that invertase and PM ATPase colocalized in secretory vesicles. These results strengthen the view that secretion and plasma membrane assembly are biosynthetically coupled in yeast. PMID- 2967287 TI - Reaction of two heads of gizzard myosin with ATP. AB - The reaction intermediates formed by the two heads of smooth muscle myosin were studied. The amount of myosin-phosphate-ADP complex, MPADP, formed was measured from the Pi-burst size over a wide range of ATP concentrations. At low concentrations of ATP, the Pi-burst size was 0.5 mol/mol myosin head, and the apparent Kd value was about 0.15 microM. However, at high ATP concentrations, the Pi burst size increased from 0.5 to 0.75 mol/mol myosin head with an observed Kd value of 15 microM. The binding of nucleotides to gizzard myosin during the ATPase reaction was directly measured by a centrifugation method. Myosin bound 0.5 mol of nucleotides (ATP and ADP) with high affinity (Kd congruent to 1 microM) and 0.35 mol of nucleotides with low affinity (Kd = 24 microM) for ATP. These results indicate that gizzard myosin has two kinds of nucleotide binding sites, one of which forms MPADP with high affinity for ATP while the other forms MPADP and MATP with low affinity for ATP. We studied the correlation between the formation of MPADP and the dissociation of actomyosin. The amount of Pi-burst size was not affected by the existence of F-actin, and when 0.5 mol of ATP per mol of myosin head was added to actomyosin (1 mg/ml F-actin, 5 microM myosin at 0 degrees C) most (93%) of the added ATP was hydrolyzed in the Pi-burst phase. All gizzard actomyosin dissociated when 1 mol of ATP per mol myosin head was added to actomyosin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967288 TI - Identification and characterization of the calmodulin-binding domain of neuromodulin, a neurospecific calmodulin-binding protein. AB - Neuromodulin (formerly designated P-57) is an abundant, neural specific, calmodulin-binding protein which exhibits higher affinity for calmodulin in the absence of free Ca2+ than in the presence of free Ca2+. In this study a series of proteolytic fragments of neuromodulin were systematically screened for calmodulin Sepharose binding activity. A 9-amino acid fragment, designated M1-C1 and having the sequence RGHITRKKL, was identified as the putative CaM-binding domain of neuromodulin. Two heptadecapeptides, designated FP57-Phe and FP57-Trp, were synthesized, each containing the M1-C1 sequence and the four flanking amino acids from each site. The FP57-Trp peptide contained a tryptophan residue in place of the native phenylalanine. Anti-FP57-Phe antibody binding to neuromodulin was inhibited by preincubation of antibodies with excess FP57-Phe. 125I-CaM gel overlay of neuromodulin was inhibited by anti-FP57-Phe antibodies. Addition of CaM to FP57-Trp increased peptide tryptophanyl fluorescence. In the presence of Ca2+, the stoichiometry of the FP57-Trp.CaM complex was 1:1, FP57-Trp binding to CaM was competitive with neuromodulin. The Ca2+-independent dissociation constant of the FP57-Phe.CaM complex was 0.41 microM. The Ca2+-dependent affinity of the complex could not be measured directly but appeared to be significantly greater than the Ca2+-independent affinity. PMID- 2967289 TI - Multiple very late antigen (VLA) heterodimers on platelets. Evidence for distinct VLA-2, VLA-5 (fibronectin receptor), and VLA-6 structures. AB - After removal of very late antigen (VLA) 2 material from a radiolabeled detergent lysate of platelets, another VLA heterodimer was precipitated using antibody to the common VLA beta subunit. This structure was identified as VLA-5 because it contained VLA beta plus an alpha subunit that was (i) recognized by anti-alpha 5 antibodies and (ii) cleaved by V8 protease to yield a characteristic alpha 5-like pattern of peptide fragments. Besides VLA-2 and VLA-5, a third heterodimer, here named VLA-6, was also present on platelets. VLA-6 (an alpha 6 beta complex) was defined using the monoclonal antibody GoH3 (Sonnenberg, A., Janssen, H., Hogervorst, F., Calafat, J., and Hilgers, J. (1987) J. Biol. Chem. 262, 10376 10383). Although it resembled VLA-5 in size, VLA-6 was different from VLA-5 because (i) removal of the alpha 5 subunit did not remove alpha 6, (ii) removal of alpha 6 by the GoH3 antibody did not remove alpha 5, (iii) the alpha 5 and alpha 6 subunits had very distinct one-dimensional V8 peptide maps, and (iv) the alpha 6 and alpha 5 subunits had distinct migration patterns on two-dimensional O'Farrell gels. The beta subunit of VLA-6 was identified as the common VLA beta subunit because (i) it was recognized by anti-VLA beta antibody and (ii) it yielded a V8 protease cleavage map characteristic of beta. VLA-6 was not readily seen in anti-VLA beta immunoprecipitations, apparently because the alpha 6 subunit is only loosely or partially associated with the VLA beta subunit. Because VLA-5 and VLA-6 both closely resemble the previously defined Ic-IIa platelet protein complex, it is likely that there is more than one platelet "Ic" protein complexed with IIa. PMID- 2967291 TI - Cloning and expression of cDNA for human endonexin II, a Ca2+ and phospholipid binding protein. AB - Endonexin II is a member of the family of Ca2+-dependent phospholipid binding proteins known as annexins. We cloned human endonexin II cDNA and expressed it in Escherichia coli. The apparent size and Ca2+-dependent phospholipid binding properties of purified recombinant endonexin II were indistinguishable from those of the placental protein. A single mRNA of approximately 1.6 kilobase pairs was found to be expressed in human cell lines and placenta and was in close agreement with the length of the cDNA clone (1.59 kilobase pairs). The cDNA predicted a 320 amino acid protein with a sequence that was in agreement with the previously determined partial amino acid sequence of endonexin II isolated from placenta. Endonexin II contained 58, 46, and 43% sequence identity to protein II, calpactin I (p36, protein I), and lipocortin I (p35), respectively. The partial sequence of bovine endonexin I was aligned with the sequence of endonexin II to give 63% sequence identity. Like these other proteins, endonexin II had a 4-fold internal repeat of approximately 70 residues preceded by an amino-terminal domain lacking similarity to the repeated region. It also had significant sequence identity with 67-kDa calelectrin (p68), a protein with an 8-fold internal repeat. Comparing the amino-terminal domains of these four proteins of known sequence revealed that, in general, only endonexin II and protein II had significant sequence identity (29%). Endonexin II was not phosphorylated by Ca2+/phospholipid-dependent enzyme (protein kinase C) even though it contained a threonine at a position analogous to the protein kinase C phosphorylation sites of lipocortin I, calpactin I, and protein II. PMID- 2967292 TI - The post-translational processing of rat pro-atrial natriuretic factor by primary atrial myocyte cultures. AB - Primary cultures of neonatal rat atrial myocytes were maintained in two different serum-free media for up to 25 days. Reversed-phase high performance liquid chromatography coupled with atrial natriuretic factor (ANF)-specific radioimmunoassay demonstrated that the cultures maintained in our previously described serum-free medium (Glembotski, C.C., and Gibson, T. R. (1985) Biochem. Biophys. Res. Commun. 132, 1008-1017) secreted primarily ANF-(1-126)-like material, whereas those cultures maintained in a different formulation of medium secreted mostly ANF-(99-126)-like material. Cultures that secreted ANF(99-126) like material were biosynthetically labeled with [35S]cysteine followed by immunoprecipitation of secreted ANF and analysis by reversed-phase, size exclusion, and ion-exchange high performance liquid chromatography. The labeled ANF-(99-126)-like peptide was shown to be chromatographically indistinguishable from other synthetic peptides related to ANF-(99-126). Labeled ANF purified from extracts of the cultured cells was chromatographically indistinguishable from authentic ANF-(1-126), and could be cleaved specifically by thrombin into labeled ANF-(99-126)-like material. These results indicate that primary atrial myocytes maintained under certain serum-free conditions are capable of secreting ANF related material that is chromatographically indistinguishable from ANF-(99-126), the known circulating form of the hormone. Additional preliminary studies suggest that the presence of glucocorticoids in the culture medium may confer ANF processing ability on cultured myocytes. PMID- 2967293 TI - Binding, internalization, and intracellular localization of interleukin-1 beta in human diploid fibroblasts. AB - This study demonstrates internalization of interleukin-1 (IL-1) via its cell surface receptor on human diploid fibroblasts and shows intracellular localization of IL-1 beta. Binding experiments at 8 degrees C using confluent fibroblast monolayers revealed 5,000-15,000 IL-1 receptors/cell that bound both IL-1 alpha and IL-1 beta. Incubation of monolayers with 125I-IL-1 beta (10(-9) M) at 8 degrees C and then at 37 degrees C for various times up to 8 h revealed a t1/2 for internalization of receptor-bound IL-1 beta of about 1.5 h. In addition, it was shown that IL-1 beta internalized via receptors was undegraded and retained binding activity. Electron microscopic autoradiography of monolayers incubated with 125I-IL-1 beta, as above, showed a progressive increase in the ratio of cytoplasmic to cell surface-associated grains. Grains at the cell surface were primarily localized at cell processes or attachment sites, frequently close to intra- and extracellular filamentous material. During incubation at 37 degrees C, most grains were free in the cytoplasm, with few present in lysosomes or vesicles. After 1 h, approximately 15% of the grains were over nuclei. Control cultures incubated at 37 degrees C with 125I-IL-1 beta and 100-fold excess unlabeled IL-1 beta showed increased uptake of label into lysosomes and little into nuclei. This study shows that IL-1 receptors are primarily located at fibroblast processes and that receptor-mediated internalization of the ligand is slow. Nuclear localization apparently requires IL-1 receptor-specific internalization of IL-1 beta, suggesting a possible role for this process in eliciting the IL-1 signal. PMID- 2967294 TI - Control of glycoprotein synthesis. Purification and characterization of rabbit liver UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N acetylglucosaminyltransferase I. AB - UDP-N-acetylglucosamine:alpha-3-D-mannoside beta-1,2-N acetylglucosaminyltransferase I catalyzes an essential first step in the conversion of high mannose to hybrid and complex N-glycans (Schachter, H. (1986) Biochem. Cell Biol. 64, 163-181; Oppenheimer, C.L., and Hill, R.L. (1981) J. Biol. Chem. 256, 799-804), i.e. the addition of GlcNAc to (Man alpha 1-6(Man alpha 1-3)Man alpha 1-6)(Man alpha 1-3)Man beta 1-4GlcNAc-OR to form (Man alpha 1 6(Man alpha 1-3)Man alpha 1-6)(GlcNAc beta 1-2Man alpha 1- 3)Man beta 1-4GlcNAc OR. The enzyme has been purified from Triton X-100 extracts of rabbit liver by chromatography on CM-Sephadex, Affi-Gel blue, UDP-hexanolamine-Sepharose, and a novel adsorbent in which UDP-GlcNAc is linked to thiopropyl-Sepharose at the 5 position of uracil. The enzyme exists in crude liver extracts in two molecular weight forms separable on Sephadex G-200. The low molecular weight form was purified 64,000-fold with a specific activity of 19.8 mumol/min/mg. The pure enzyme was free of N-acetylglucosaminyltransferase II-V activities. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single major band of Mr 45,000 and two minor bands of Mr 54,000 and 50,000. All three bands showed retarded elution from an affinity column in which the acceptor substrate for the transferase was covalently linked to Sepharose. Kinetic analysis indicated a largely ordered sequential mechanism with UDP-GlcNAc binding to the enzyme first and UDP leaving last. Studies with synthetic analogues of the substrate Man alpha 1-6(Man alpha 1-3)Man beta 1-4GlcNAc showed that an unsubstituted equatorial hydroxyl on carbon 4 of the beta-linked Man residue was essential for enzyme activity. PMID- 2967290 TI - Transient inhibition of DNA synthesis by 5-fluorodeoxyuridine leads to overexpression of dihydrofolate reductase with increased frequency of methotrexate resistance. AB - Transient but incomplete suppression of DNA synthesis by a single exposure of an asynchronous population of cells to 5-fluoro-2'-deoxyuridine (FdUrd) increases the frequency of appearance of methotrexate (MTX)-resistant colonies. This increase was greater than 10-fold following a 6-h incubation of cells with 3 microM FdUrd prior to selection in MTX, an interval one-half the normal L1210 cell cycle time. During this period of exposure to FdUrd, DNA synthesis decreased to 25% of control rates and cells accumulated at the G1/S interface. The 6-h incubation with FdUrd resulted in greater than a 2.5-fold increase in the dihydrofolate reductase protein level in the treated cell population, which was accounted for, at least in part, by increased de novo synthesis of the enzyme as assessed by [35S]methionine labeling. This increase in dihydrofolate reductase was associated with a decrease in growth inhibition by MTX. A brief reversal (2 h) of FdUrd-induced DNA synthesis inhibition by the addition of thymidine eliminated the amplification of dihydrofolate reductase and the enhanced emergence of MTX-resistant clones. Beyond this, an analysis of clones that survive MTX selection indicates that the dihydrofolate reductase gene copy in cells spontaneously resistant to 50 nM MTX and those which resulted after the additional pretreatment with FdUrd for 6 h are comparable with a 2-4-fold amplification of enzyme in most clones. These studies demonstrate that FdUrd enhancement of dihydrofolate reductase expression can have a profound effect upon the incidence and expression of MTX resistance and that dihydrofolate reductase gene amplification may be another basis for antagonism between these agents. PMID- 2967295 TI - The Nu1 subunit of bacteriophage lambda terminase. AB - The maturation and packaging of bacteriophage lambda DNA are catalyzed by the phage terminase enzyme. Terminase is composed of two protein subunits, gpNu1 and gpA. The holoenzyme is multifunctional in vitro; it binds to and cleaves lambda DNA at the cos site (where cos represents cohesive-end site), packages DNA into lambda proheads, and is also a DNA-dependent ATPase. The genes of the two subunits have been cloned separately into powerful expression vectors which allow for very high levels of protein overproduction. The gpNu1 protein has been purified to homogeneity and has a monomeric molecular weight of 21,200, in close agreement with the Mr of 20,444 expected from its amino acid sequence. Both gel filtration and sedimentation velocity centrifugation indicate that the native gpNu1 protein exists as a Mr greater than 500,000 aggregate. The sequence of the first 20 amino acids and the overall composition both match those predicted by the nucleotide sequence of the Nu1 gene. Purified gpNu1 is able to complement gpA containing extracts in both lambda DNA packaging and cos cleavage assays. The Nu1 gene amino acid sequence predicts DNA binding by the protein, and gpNu1 does show specific binding to lambda DNA by filter binding assays. Also, as predicted from its sequence, gpNu1 exhibits ATPase activity; but in contrast to the holoenzyme, this activity is DNA-independent. PMID- 2967297 TI - Cell morphology, proliferation and collagen synthesis of human fibroblasts cultured on sepiolite-collagen complexes. AB - The growth and morphology as well as collagen biosynthesis of human fibroblasts obtained and cultured on sepiolite-collagen complexes have been studied. No differences on cell morphology and growth properties nor collagen synthesis were observed when compared with standard culture substrates. The type I/type III ratio of biosynthesized collagen by fibroblasts cultured on sepiolite-collagen complexes was about 5-6 with no difference when compared to control conditions. This normal behavior was also observed for the type I/type III procollagens. According to these studies the sepiolite-collagen complexes do not modify the studied features of the fibroblasts. PMID- 2967296 TI - Transfection of genes encoding the T cell receptor-associated CD3 complex into COS cells results in assembly of the macromolecular structure. AB - The T cell antigen receptor (TCR) consists of a disulfide-linked TCR-alpha/beta heterodimer that is both structurally and functionally associated with a set of four non-covalently linked membrane proteins termed CD3-gamma, -delta, -epsilon, and -zeta. An additional protein described recently, CD3-omega, has been suggested to play a role in assembly of the CD3 complex on the basis of its transient association with the CD3 proteins early during biosynthesis. Association of all the proteins seems to be a prerequisite for intracellular transport, since mutants lacking either the TCR-alpha or -beta protein do not express the CD3 complex on the cell surface. CD3-cDNAs were transfected into COS cells in order to study the protein-protein interactions ruling the assembly of the CD3 macromolecular structure. CD3-delta-epsilon, CD3-gamma-epsilon, and CD3 gamma-delta-epsilon intermediates could be detected. These data indicated that a CD3 core structure could be formed in the absence of the other members of the complex (CD3-zeta, -omega, TCR-alpha, and -beta). Both the individual CD3 chains and the assembled CD3.gamma.delta.epsilon complexes could not be detected on the cellular surface but in an intracellular compartment, probably the endoplasmic reticulum or the cis Golgi. The transfection experiments allowed us to identify the 25-kDa member of the murine CD3 complex as CD3-epsilon m. Furthermore, a 23 kDa glycoprotein seen upon metabolic labeling of human T cells was shown to be an immature form of the CD3-gamma h protein. PMID- 2967298 TI - Surgery for malignant extradural tumours of the spine. AB - We have reviewed 41 patients with malignant extradural tumours of the spine treated by anterior decompression for cord compression, or uncontrolled back pain or both. An anterior operation alone was performed in 37 cases, four had combined or staged anterior and posterior decompression. An anterior operation on its own achieved major neurological recovery in 18 of the 33 cases with neurological loss (56%); only four remained unchanged. Eleven had minor improvement but not enough to allow them to walk or to regain bladder function. No patient with complete paraplegia gained a useful neurological recovery. Back pain was improved in 30 of the 41 patients (73%), sound internal fixation being important in this respect. There were four early deaths and another 23 died from disseminated disease after a mean survival of 4.1 months. Fourteen patients are still alive with a mean survival of 14 months. PMID- 2967299 TI - Proteolytic activation of latent transforming growth factor-beta from fibroblast conditioned medium. AB - Transforming growth factor-beta (TGF beta) is produced by most cultured cells in an inactive form. Potential activation mechanisms of latent TGF beta were studied using fibroblastic (NRK-49F and AKR-MCA) cell-conditioned medium as a model. Active TGF beta was monitored by radioreceptor and soft agar assays as well as by antibody inhibition and immunoprecipitation. Little or no TGF beta was detected in untreated conditioned medium. Treatment of the medium with extremes of pH (1.5 or 12) resulted in significant activation of TGF beta as shown by radioreceptor assays, while mild acid treatment (pH 4.5) yielded only 20-30% of the competition achieved by pH 1.5. In an effort to define more physiological means of TGF beta activation, the effects of some proteases were tested. Plasmin and cathepsin D were found to generate 25-kD bands corresponding to the active form of TGF beta as shown by immunoprecipitation analysis of radiolabeled cell-conditioned medium. Plasmin treatment of the medium resulted in activity that was quantitatively similar to that of mild acid treatment as measured by radioreceptor and soft agar assays. In addition, the plasmin-generated activity was inhibited by anti-TGF beta antibodies. Sequential treatments of AKR-MCA cell-conditioned medium with mild acid followed by plasmin or plasmin followed by mild acid gave activation comparable to either treatment alone. The data suggest that conditioned medium may contain at least two different pools of latent TGF beta. One pool is resistant to mild acid and/or plasmin and requires strong acid or alkali treatment for activation. A second pool is activated by mild pH change and/or plasmin. Activation of this form of latent TGF beta may take place by dissociation or proteolytic digestion from a precursor molecule or hypothetical TGF beta-binding protein complex. PMID- 2967300 TI - Distribution of a 69-kD laminin-binding protein in aortic and microvascular endothelial cells: modulation during cell attachment, spreading, and migration. AB - Affinity chromatography and immunolocalization techniques were used to investigate the mechanism(s) by which endothelial cells interact with the basement membrane component laminin. Bovine aortic endothelial cells (BAEC) membranes were solubilized and incubated with a laminin-Sepharose affinity column. SDS-PAGE analysis of the eluted proteins identified a 69-kD band as the major binding protein, along with minor components migrating at 125, 110, 92, 85, 75, 55, and 30 kD. Polyclonal antibodies directed against a peptide sequence of the 69-kD laminin-binding protein isolated from human tumor cells identified this protein in BAEC lysates. In frozen sections, these polyclonal antibodies and monoclonal antibodies raised against human tumor 69-kD stained the endothelium of bovine aorta and the medial smooth muscle cells, but not surrounding connective tissue or elastin fibers. When nonpermeabilized BAEC were stained in an in vitro migration assay, there appeared to be apical patches of 69 kD staining in stationary cells. However, when released from contact inhibition, 69 kD was localized to ruffling membranes on cells at the migrating front. Permeabilized BAEC stained for 69 kD diffusely, with a granular perinuclear distribution and in linear arrays throughout the cell. During migration a redistribution from diffuse to predominanately linear arrays that co-distributed with actin microfilaments was noted in double-label experiments. The 69-kD laminin-binding protein colocalized with actin filaments in permeabilized cultured microvascular endothelial cells in a continuous staining pattern at 6 h postplating which redistributed to punctate patches along the length of the filaments at confluence (96 h). In addition, 69 kD co-distribution with laminin could also be demonstrated in cultured subconfluent cells actively synthesizing matrix. Endothelial cells express a 69-kD laminin-binding protein that is membrane associated and appears to colocalize with actin microfilaments. The topological distribution of 69 kD and its cytoskeletal associations can be modulated by the cell during cell migration and growth suggesting that 69 kD may be a candidate for a membrane protein involved in signal transduction from extracellular matrix to cell via cytoskeletal connections. PMID- 2967301 TI - Heparan sulfate proteoglycans in the substratum adhesion sites of human neuroblastoma cells: modulation of affinity binding to fibronectin. AB - Tissue culture substratum adhesion sites from EGTA-detached Platt human neuroblastoma cells were extracted with a buffer containing ocytlglucoside, NaCl, guanidine hydrochloride, and a variety of protease inhibitors, an extraction which resulted in quantitative solubilization of the 35SO4 = -radiolabeled proteoglycans and 3H-leucine-radiolabeled proteins. Of the sulfate-radiolabeled material, the vast majority was heparan sulfate proteoglycan (Kav = 0.15 on Sepharose C14B columns) and the remainder was chondroitin sulfate chains (no single chains of heparan sulfate were observed). This extract was then fractionated on DEAE-Sephadex columns under two different buffer elution conditions. Under DEAE-I conditions in low ionic strength acetate buffer, two major peaks of 35SO4 = -radiolabeled material (A,B) and a minor peak (C) could be resolved in the NaCl gradient; however, three-fourths of the material required 4 M guanidine hydrochloride to elute it from the column (peak D). Under DEAE-II conditions in acetate buffer supplemented with 8 M urea, the vast majority of the proteoglycan material could be eluted in the NaCl gradient as peak AB. Peak D material was shown to contain aggregated proteoglycan, along with nonproteoglycan protein, which high concentrations of urea or guanidine could dissociate, but not nonionic or zwitterionic detergents. Three different affinity chromatography systems were used to further characterize these components. Approximately 60% of peak A heparan sulfate proteoglycan from DEAE-I binds to the hydrophobic matrix, octyl-Sepharose, while 80% of the proteoglycan in DEAE-I peak D binds to this hydrophobic column. A sizable fraction of peak A proteoglycan fails to bind to plasma fibronectin but does bind to platelet factor-4 affinity columns. In contrast, peak AB proteoglycan from DEAE-II columns yields a much higher proportion of molecules which do bind to fibronectin. To examine the basis for these differences in affinity binding, nonproteoglycan protein from these adhesion sites was mixed with peak AB proteoglycan prior to affinity chromatography; proteoglycan binding to fibronectin decreased markedly while binding to platelet factor-4 was unaffected. This modulating activity involves the binding of nonproteoglycan protein in adhesion site extracts to both fibronectin on the column, as well as to heparan sulfate proteoglycan itself, and it could not be mimicked by a number of known proteins in adhesion site extracts or several other proteins. These results demonstrate selectivity and specificity in this modulation and indicate that a previously unidentified protein(s) is responsible.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967302 TI - Sulfated glycosaminoglycans modify growth factor-induced neurite outgrowth in PC12 cells. AB - Glycosaminoglycans (GAGs), localized on the surfaces of cells and in the basement membrane, modulate the growth and differentiation of many cell types. Recent studies have shown that heparin, a GAG found in mast cells, potentiates the ability of acidic fibroblast growth factor (aFGF) to induce neurite outgrowth in pheochromocytoma (PC12) cells. We examined the effect of a variety of GAGs on aFGF, basic fibroblast growth factor (bFGF), and nerve growth factor (NGF) induced neurite outgrowth in PC12 cells. The effects observed were dependent upon the specific GAG, the concentration of the GAG, and the growth factor. Heparin potentiated aFGF-induced neurite outgrowth in a concentration-dependent fashion; potentiation increased with increasing heparin concentrations of 0.01-100 micrograms/ml. At concentrations greater than 100 micrograms/ml, heparin potentiation decreased. The maximally active concentration of heparin (100 micrograms/ml) increased the potency of aFGF 102-fold. Increasing concentrations of heparan sulfate, dermatan sulfate, and chondroitin sulfate correlated with increasing aFGF potentiation. The maximally active concentrations of heparan sulfate (100 micrograms/ml), dermatan sulfate (10 mg/ml), and chondroitin sulfate (1 mg/ml) increased the activity of aFGF 11-, 110-, and 11-fold, respectively. Hyaluronic acid did not affect the neurite outgrowth-promoting activity of aFGF. Heparin also altered the activity of bFGF; increasing concentrations of heparin (0.01-1 micrograms/ml) correlated with increased potentiation. At concentrations greater than 1 microgram/ml, heparin concentration was inversely correlated with potentiation. Chondroitin sulfate only increased the percentage of neurite bearing cells at concentrations greater than 10 micrograms/ml. Maximally active concentrations of heparin (1 microgram/ml) and chondroitin sulfate (1 mg/ml) increased the potency of bFGF 5-fold. The highest concentration of heparan sulfate studied (1 mg/ml) inhibited the activity of bFGF. Dermatan sulfate and hyaluronic acid (0.01-1000 micrograms/ml) had no effect on bFGF activity. Heparan sulfate and chondroitin sulfate showed concentration-dependent potentiation of NGF; maximally active concentrations of heparan sulfate (100 micrograms/ml) and chondroitin sulfate (1 mg/ml) increased the potency of NGF 3-fold, whereas heparin, dermatan sulfate and hyaluronic acid had no effect. None of the GAGs had any effect on PC12 neurite outgrowth when added alone. The specificity of the activity of the GAGs was verified by selective enzyme degradation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967303 TI - Dyslipidemia and accelerated atherosclerosis. PMID- 2967305 TI - Divergent correlations of circulating dehydroepiandrosterone sulfate and testosterone with insulin levels and insulin receptor binding. AB - We evaluated the insulin response to a standard oral glucose tolerance test (OGTT) and in vitro insulin binding to erythrocytes (RBC) in 26 women from 3 groups: Group NW, normal women (n = 11); Group DS, women (n = 9) with elevated serum DHEAS concentrations, greater than 400 micrograms/dl (greater than 10.84 mumol/L); and Group IR, women (n = 6) with elevated basal plasma insulin concentrations (IRI). There was a significant linear correlation between the area under the insulin response curve (IRI-AUC) and serum testosterone (T) (r = 0.78, p = 0.0001). Using stepwise multiple linear regression, IRI-AUC was characterized as a function of both serum T and DHEAS; positively with T and negatively with DHEAS. In vitro (n = 17), there was a positive correlation between RBC-insulin binding and serum DHEAS (r = 0.54, p = 0.029) and a negative correlation between RBC-binding and T (r = -0.57, p = 0.017). We conclude that DHEAS may enhance insulin binding and action and that DHEAS and T have divergent functional relationships with IRI. DHEAS and T may therefore exert opposing effects on insulin secretion and action. PMID- 2967304 TI - Glucocorticoid and mineralocorticoid stimulation of atrial natriuretic peptide release in man. AB - To investigate the influence of a mineralocorticoid and a glucocorticoid on plasma immunoreactive atrial natriuretic peptide (irANP) and possible functional correlates, eight normal men received in random order 9 alpha-fludrocortisone acetate (9 alpha F; 0.6 mg/day), prednisone (50 mg/day), and placebo each for 9 days. Their diet contained 130 mmol sodium and 75 mmol potassium daily. The mean supine plasma irANP levels were similar on days 2, 4, and 9 of placebo treatment [25 +/- 10 (+/- SE), 27 +/- 5, and 27 +/- 6 pmol/L, respectively]. Mean plasma irANP levels were 76 +/- 42 (P less than 0.05), 89 +/- 34, and 93 +/- 29 pmol/L (P less than 0.01), respectively, on days 2, 4, and 9 during 9 alpha F administration, and 68 +/- 37 (P less than 0.05), 83 +/- 41, and 48 +/- 18 pmol/L on the same days during prednisone administration. Compared with the placebo period, sodium intake minus urinary output during 9 alpha F administration averaged +41 mmol at the time of blood sampling on day 2, +112 mmol on day 4, and +149 mmol on day 9; body weight was unchanged on day 2 and increased by 0.7 and 1.1 kg on days 4 and 9, respectively. Escape from 9 alpha F-induced renal sodium retention occurred on days 5 and 6. During prednisone administration, sodium intake minus urinary output and body weight did not change. Plasma volume and BP rose significantly during 9 alpha F (P less than 0.05) but not during prednisone administration. Plasma renin, aldosterone, and norepinephrine (NE) decreased during 9 alpha F treatment (P less than 0.05 to less than 0.01); during prednisone treatment, plasma aldosterone levels were lower on day 9 only. Cardiovascular pressor responsiveness to angiotensin II was enhanced during 9 alpha F but not prednisone administration, while blood pressure reactivity to NE was not significantly modified. These findings demonstrate that 9 alpha F and prednisone in high doses provoke remarkably similar increases in plasma irANP, but that the glucocorticoid-induced rise in plasma irANP is due to a mechanism other than sodium and volume retention. PMID- 2967306 TI - In vitro activation of T lymphocytes from human immunodeficiency virus (HIV) seropositive blood donors. I. Soluble interleukin 2 receptor (IL2R) production parallels cellular IL2R expression and DNA synthesis. AB - We investigated the relationship of soluble interleukin 2 receptor (sIL2R) production to cellular IL2R expression and DNA synthesis by mitogen-stimulated mononuclear cells from blood donors seropositive for human immunodeficiency virus (HIV). SIL2R was measured using an enzyme-linked immunosorbent assay which employed 2 anti-IL2R monoclonal antibodies recognizing distinct IL2R epitopes. Decreased phytohemagglutinin-induced DNA synthesis and cellular IL2R expression were accompanied by decreased levels of sIL2R in cell culture supernatants. Similar findings were observed for pokeweed mitogen-induced responses. There was no detectable spontaneous secretion of sIL2R into culture supernatants by unstimulated mononuclear cells from either HIV-seropositive or control seronegative donors. These findings indicate that the in vitro T-cell activation defects which characterize HIV infection include decreased sIL2R production, as well as decreased cellular IL2R expression and DNA synthesis. Further, they show that assessment of supernatant sIL2R levels can be used as a valid, reliable assay for T-cell activation. PMID- 2967307 TI - Natural killer-cell activity in cyclosporine-treated renal allograft recipients. AB - We prospectively studied natural killer (NK)-cell activity in 16 cyclosporine treated renal transplant recipients. NK function remained intact in the group as a whole in the initial 6 months following transplantation. The percentage of CD16 positive cells within the peripheral blood mononuclear-cell population was highly correlated with NK activity both prior to and following transplantation in the absence of rejection. During rejection, the correlation was poor. A marked increase in NK activity occurred during 9 of 12 rejection episodes; similar increases in NK activity were rarely observed in the absence of rejection. Significant infiltrates of NK cells, as determined by expression of CD16, were not demonstrated in stained biopsy specimens obtained from rejecting allografts. Pretransplant NK activity did not predict clinical outcome of the allograft. Our results indicate that NK cells are activated during allograft rejection in cyclosporine-treated patients, but their exact role in the rejection process is unknown. PMID- 2967308 TI - Functional analysis of CD8 lymphocytes in long-term surviving patients after bone marrow transplantation. AB - The recovery of T-cell populations after bone marrow transplantation (BMT) is characterized by a persistent expansion of CD8 lymphocytes. Previously, we have shown that beyond 1 year posttransplantation the CD8 lymphocytes consist, to a large extent, of CD8+ HNK1+ cells that suppress, like normal CD8 lymphocytes, immunoglobulin production in vitro. We have further investigated the functional capabilities of CD8 lymphocytes, mostly HNK1+ (from 50 to 77%), in seven long term BMT patients. As normal, patient CD8 lymphocytes do not suppress (1) phytohemagglutinin (PHA)-induced interleukin 2 (IL2) receptor expression and IL2 responsiveness by normal T cells or (2) the mixed lymphocyte reaction of donor cells. Also as normal, patient CD8 lymphocytes can be activated into potent cytotoxic effectors. Therefore, under the present experimental conditions, the increase in the absolute number of CD8 lymphocytes in the long-term BMT patients is characterized by an expansion of the CD8+ HNK1+-cell subpopulation and a normal suppressor/cytotoxic potential on a per-CD8+ cell basis. PMID- 2967309 TI - Methionine-enkephalin as immunomodulator therapy in human immunodeficiency virus infections: clinical and immunological effects. AB - Enkephalins have been shown to enhance T cell-mediated immune responses and natural killer-cell activity in vitro. We have studied the effects of infusions of methionine-enkephalin on immune functions and clinical courses in seven patients with various stages of infection with human immunodeficiency virus (HIV). All patients were clinically stable at the time of entry into the study. Each received 10 micrograms/kg of methionine-enkephalin in an intravenous infusion three times weekly for up to 12 weeks. Evaluation of cellular immunity (T-cell subsets, in vitro interleukin-2 production and interleukin-2 receptor expression, T-cell responses to mitogens and antigens, and delayed hypersensitivity skin tests) as well as clinical and toxicity monitoring was performed prior to treatment, at 2-week intervals during treatment, and after the cessation of treatment. Increases in interleukin-2 receptor expression were seen on lymphocytes collected on one occasion from each of two patients 30 min postinfusion. Studies done 24 hr after infusions revealed increases in interleukin-2 production in one patient, but when pre- and posttreatment values were compared there were no significant changes in numbers of circulating T cells of any phenotype or in T-cell responses to mitogens or antigens. None of the patients with Kaposi's sarcoma had regression of tumor; one patient dropped out of the study at week 5 because of deteriorating clinical status and progression of tumor. There were no adverse reactions or evidence of toxicity. We conclude that methionine-enkephalin appears to enhance temporarily selected immune responses in patients with HIV infection, however, in the schedule used in this study it was not clinically efficacious. PMID- 2967310 TI - Acitretin improves psoriasis in a dose-dependent fashion. AB - Acitretin, a metabolite of etretinate, was given to 38 patients for the treatment of psoriasis. During the first 8 weeks patients received either placebo, 10 mg, 25 mg, 50 mg, or 75 mg of acitretin daily in a double-blind manner. The dosages of 10 mg and 25 mg daily did not achieve any statistically significant improvement in psoriasis over placebo; however, both the 50 and 75 mg dosages were statistically significantly better than placebo. Side effects were primarily mucocutaneous and occurred in most patients receiving 25 mg or more of acitretin daily. After the double-blind period, patients continued treatment in an open fashion until they had received a total of 24 weeks of acitretin therapy. Most patients received 50 mg of acitretin daily, which adequately cleared their psoriasis. After approximately 3 months without acitretin, most patients required retreatment. Subsequent 24-week courses of therapy were generally effective and well tolerated. The most common laboratory abnormalities were elevations of triglyceride, cholesterol, and liver transaminase levels. The efficacy and side effects of acitretin appear to be similar to those of etretinate; the principal advantage of acitretin is its shorter half-life. Although acitretin is a potent teratogen, its rapid elimination makes it a viable treatment for psoriasis among women of childbearing potential. PMID- 2967311 TI - Cutaneous and immunologic reactions to phenytoin. AB - Phenytoin (diphenylhydantoin; Dilantin) is a highly effective and widely prescribed anticonvulsant and antiarrhythmic agent. Since 1938 it has been invaluable in the treatment of grand mal and psychomotor epilepsy. Hydantoin derivatives have been used medicinally for more than a half-century. In recent years dermatologists have broadened the indications for phenytoin use to include recessive dystrophic epidermolysis bullosa, linear scleroderma, and pachyonychia congenita. In spite of widespread use and popularity, it is interesting that the frequency of complications relating to drug therapy remains low, relatively speaking. Nevertheless, a broad spectrum of cutaneous and immunologic reactions to phenytoin have been reported. These range from tissue proliferative syndromes (side effects), drug hypersensitivity syndromes (allergic effects), and a possible linkage with lymphoma (idiosyncratic effects). Therapeutic and toxic reactions to this commonly prescribed drug are comprehensively reviewed, analyzed, and summarized in this monograph. PMID- 2967312 TI - Cystic acne in psoriatic patients undergoing etretinate therapy. PMID- 2967313 TI - Phagocytic activity of milk leukocytes during chronic staphylococcal mastitis. AB - Receptors for IgG on milk leukocytes were detected by rosette formation using sensitized erythrocytes. The percentage (41) of milk leukocytes from uninfected glands forming sensitized erythrocyte rosettes was significantly greater than the percentage (13) of leukocytes from glands with chronic staphylococcal mastitis. A greater percentage of polymorphonuclear leukocytes than macrophages formed sensitized erythrocyte rosettes, regardless of the infection status of the gland from which they were obtained. Both IgG-receptor and nonimmunologic receptor mediated phagocytosis were greater for milk leukocytes from uninfected glands than for milk leukocytes from chronically infected glands. Preincubation of normal milk leukocytes in whey prepared from mastitic milk resulted in a decrease in their capacity to form sensitized erythrocyte rosettes as well as a reduction in their phagocytic capacity. Immune complexes prepared in vitro also reduced the phagocytic capacity of normal milk leukocytes and inhibited their capacity to form sensitized erythrocyte rosettes. These data indicate a factor, possibly consisting of immune complexes, was present in secretions from glands chronically infected with staphylococci. This factor reduced the phagocytic capacity of milk leukocytes. PMID- 2967314 TI - Comparison of operant behavioral and cognitive-behavioral group treatment for chronic low back pain. PMID- 2967315 TI - Role of chemical mediators after antigen and exercise challenge in children with asthma. AB - Changes in chemical mediators after antigen challenge and exercise challenge tests were studied in children with asthma. Chemical mediators studied after antigen challenge and exercise challenge included histamine, leukotrienes (LTB4, LTC4, and LTD4), and neutrophil chemotactic factor of anaphylaxis (NCA). The pharmacologic modification of immediate and late-phase reactions was evaluated for procaterol (beta 2-agonist), cromolyn sodium, and prednisolone. Histamine levels were noted to rise in patients who had a dual response of a mild to moderate nature, but did not change in patients who had severe asthma. During exercise challenge cromolyn sodium inhibited both immediate and late-phase reactions but also inhibited plasma generation of NCA. Prednisolone, on the other hand, did not affect immediate reactions, but blocked late-phase reactions to exercise and also decreased NCA generation. Procaterol inhibited the generation of LTD4 and also inhibited NCA when compared with placebo. Exercise challenge did not alter levels of plasma histamine or of LTB4 or LTC4. PMID- 2967316 TI - Clinical evaluation of the plasma levels of immunoreactive atrial natriuretic peptide in elderly patients with heart diseases. AB - Plasma levels of immunoreactive atrial natriuretic peptide (ANP) were estimated in 69 elderly patients over 60 years of age (mean 76.4 years) with or without heart diseases and in ten young, healthy volunteers (mean 33.0 years) to evaluate the clinical significance of ANP in the elderly. Plasma ANP levels in nine patients without heart diseases were significantly (P less than .01) higher than in the ten young, healthy subjects (mean +/- SD, 46.0 +/- 22.0 vs 22.1 +/- 6.3 pg/mL) and a significant positive correlation was observed between ANP level and age in these subjects (r = 0.60, P less than 0.01). Plasma ANP levels in 60 patients with heart diseases (158.4 +/- 158.5 pg/mL) were significantly (P less than 0.05) greater than in nine patients without heart diseases. Plasma ANP levels in patients with congestive heart failure or atrial fibrillation were 285.8 +/- 185.2 or 223.0 +/- 185.9 pg/mL, respectively; each of these values was significantly (P less than 0.01) higher than in patients without heart diseases. In three patients with paroxysmal atrial fibrillation, plasma ANP levels during atrial fibrillation were three times greater than when atrial fibrillation returned to normal sinus rhythm (377.3 +/- 78.5 vs 101.1 +/- 68.5 pg/mL). These results indicate that plasma ANP levels increase with advancing age, and that increased ANP levels are associated with various heart diseases in elderly subjects, possibly through stretch of the atrial wall. PMID- 2967317 TI - Corticosteroids modulate the binding of recombinant interferons alpha and gamma in Namalva cells. AB - To investigate possible mechanisms of interaction between corticosteroids and interferons (IFNs), the specific binding of recombinant human IFNs alpha 2 and alpha in Namalva cells after 72 h culture with dexamethasone (10(-8) M to 10(-6) M) was evaluated. Exponentially growing cells were incubated with different concentrations of the radiolabelled IFNs, with or without an excess of unlabelled IFN. The parameters of the interaction between each IFN and its specific receptor were analyzed by the Scatchard method. In the dose range tested, dexamethasone induced a dose-dependent inhibition of Namalva cells growth, which reached about 35% at 10(-6) M. The specific binding of IFN-alpha 2 was decreased to a maximum of 40%, for dexamethasone concentrations greater than or equal to 10(-7) M. The decrease in binding induced by the corticoid was additive with the down regulation induced by IFN-alpha 2 itself. On the contrary, the specific binding of IFN-alpha was increased by dexamethasone in a dose-dependent fashion within the tested range. The maximal increase in the number of sites per cell was about 60%, with a slight decrease in affinity. These results suggest that complex interactions might arise between corticosteroids and IFNs in the course of their clinical use. PMID- 2967319 TI - [Ovarian cysts and celioscopy. Apropos 226 cases]. AB - Tumours of the adnexae are frequent. In most cases it is necessary to look at them directly and to treat them surgically. Laparatomy is a heavy technique. For this reason we use the laparoscope to diagnose cystic lesions and to treat benign tumours. This retrospective study on 226 patients has given us the opportunity to assess the value of the laparoscope in these conditions. From the diagnostic point of view laparoscopy is reliable. No malignant tumour failed to be diagnosed (100% reliable), the anatomopathological examination of specimens in benign conditions was never wrong (100% reliable). Laparoscopy therefore is a reliable way of diagnosing the type of ovarian cyst. 180 patients were treated using the laparoscope (79.5%). The patients were followed up for 1 to 5 years. We have not had a recurrence of a cyst treated in this way. Each time a malignant tumour was diagnosed or even suspected the classical surgical approach by laparotomy was always carried out. For a trained surgeon laparoscopy seems to be a reliable method of diagnosing and treating. It possesses the very well-known advantages of minimal operative trauma and far less cost. PMID- 2967320 TI - [Analogs of GnRH and fertilization in vitro. Value of the use of GnRH analogs in the stimulation and monitoring of ovulation for fertilization in vitro]. AB - Gn RH analogues seemed to us to be very useful in in-vitro fertilisation. They make it easier to programme oocyte recovery. They avoid an endogenous surge of LH, and they particularly seemed to increase significantly the percentage of pregnancies that continue after embryo transfer. PMID- 2967318 TI - Acromegalic cardiomyopathy. An echocardiographic study. AB - Eighteen acromegalic patients (A) and 18 controls without clinical evidence of cardiac involvement and/or endocrine disease (C), matched for sex, age, body surface area, and blood pressure (BP), were investigated by M-mode (2-D derived) echocardiography, to clarify the prevalence and the possible determinants of left ventricular hypertrophy (LVH). Seven patients in each group were hypertensive (BP greater than 160/95 mmHg). Left ventricular mass (LVM) was 183.1 +/- 60.0 g/m2 in A and 130 +/- 25.9 g/m2 in C. A LVM above 140 g/m2 (that is the upper normal range in our laboratory) was found in 15/18 A and 2/18 C. The LVH was concentric (h/r greater than 0.45) in 12/15 A and 1/2 C. Systolic function indexes (% FS, end-systolic stress/end-systolic volume), cardiac index and total peripheral resistance index (as determined by echo) were within the normal range and similar in both groups. No correlation was found between LVM and BP, LVM and GH plasma levels, LVM and Sm-C levels. A significant correlation was found between LVM and duration of the disease (r 0.44; p less than 0.05). Our data confirm that LVH is an early and frequent finding in acromegaly. Its prevalence is not entirely accounted for by such factors as body size, BP or increased cardiac output. Metabolic factors may play a major role, and a long lasting exposition to increased GH levels seems the most relevant determinant of LVH. PMID- 2967321 TI - [Intrauterine growth retardation and the decision factors in its management: role of Doppler velocimetry]. PMID- 2967322 TI - Teicoplanin--a new agent for gram-positive bacterial infections. PMID- 2967323 TI - Practical applications of decision analysis. AB - The number and complexity of available tests and therapies is expanding and financial resources are shrinking. Clinicians must now consider how to give the best care while limiting health expenses. Decision analysis is a tool used to guide clinicians in the practice of medicine, and to supplement and enhance intuitive decision making based on clinical experience. PMID- 2967324 TI - T cell-dependent activation of B cell proliferation and differentiation by immobilized monoclonal antibodies to CD3. AB - The capacity of mAb directed at the CD3 molecular complex (64.1) to induce T cell dependent B cell proliferation and differentiation was examined. Coculture of B cells with mitomycin C-treated T4 cells (T4 mito) stimulated by immobilized 64.1 resulted in marked B cell proliferation and Ig-secreting cells (ISC) generation in the absence of any additional stimulation. The magnitude of the B cell responses induced by immobilized 64.1-stimulated T4 mito was far greater than that induced by other stimuli, such as Staphylococcus aureus plus factors produced by mitogen-activated T cells, PWM, or soluble 64.1. The induction of maximal B cell responsiveness required direct contact between activated T cells and responding B cells. Of note, immobilized 64.1 also induced B cell proliferation and ISC generation in the presence of mitomycin C-treated T8 cells. By contrast, immobilized 64.1 stimulated T4 or T8 cells that had not been treated with mitomycin C induced very modest ISC generation and suppressed B cell responses supported by T4 mito even in the presence of exogenous IL-2 or factors produced by mitogen-activated T cells. The interactions between T and B cells in these cultures not only induced B cell responses, but also enhanced the production of IL-2 by activated T cells. Increased IL-2 production was facilitated when culture conditions afforded the opportunity for contact between B cells and activated T cells. These results indicate that the establishment of interactions between B cells and anti-CD3-stimulated T4 or T8 cells provides all of the signals necessary for proliferation and differentiation of B cells without other stimuli and also augments the production of lymphokines by the activated T cells. The data emphasize the role of Ag-nonspecific interactions between B cells and T cells in promoting polyclonal responses of both cell types. PMID- 2967325 TI - Suppressor T cells and self-tolerance. Active suppression required for normal regulation of anti-erythrocyte autoantibody responses in spleen cells from nonautoimmune mice. AB - Spleen cells from young, nonautoimmune strains of mice cultured with syngeneic E do not develop a significant anti-mouse E response in vitro, consistent with a state of self-tolerance to this Ag. In order to study the role of active suppression in regulating mouse RBC-(MRBC) specific cells in nonautoimmune cell populations, the effect of depleting T cell subsets on the generation of anti MRBC autoantibodies by nonautoimmune spleen cells was determined. Spleen cells from young BALB/c and C57BL/6 mice were found to generate significant numbers of IgM and IgG anti-MRBC autoantibody-forming cells in culture with MRBC after depletion of Ly-2+ cells by anti-Ly-2 and C treatment. The response which develops is Ag dependent, Ag specific, and dependent upon L3T4+ Th. The magnitude and isotype of this response is similar to the anti-MRBC response generated by spleen cells from 12-mo-old, autoimmune NZB mice and young NZB mice also treated to remove Ly-2+ cells. Addition of isolated Ly-2+ T cells, but not L3T4+ or Ly-2- T cells, to spleen cells depleted of Ly-2+ cells restores apparently normal regulation of the anti-MRBC response in vitro. These data demonstrate that control of a specific autoantibody response to MRBC by nonautoimmune spleen cell populations requires active regulation by an Ly-2+ T cell subset. PMID- 2967326 TI - Characterization of murine IL-1 beta. Isolation, expression, and purification. AB - One cDNA clone encoding a truncated murine IL-1 beta (M IL-1 beta) sequence was isolated from a murine macrophage cDNA library. We reconstituted the coding sequence of the 152-residue mature protein and expressed it in Escherichia coli. rM IL-1 beta was purified to homogeneity and characterized by oligonucleotide and NH2-terminal sequence analysis. Purified rM IL-1 beta exhibited biologic activity equivalent to 7.8 x 10(7) units/mg in the murine thymocyte proliferation assay and 9.9 x 10(3) units/mg in the human gingival fibroblast PGE2 production assay, indicative of species specificity. The isoelectric point of rM IL-1 was found to be 8.85. The circular dichroism spectrum revealed that the secondary structure of M IL-1 is indistinguishable from that of the human protein. Receptor binding studies indicated the rM IL-1 bound to murine EL-4.1 thymoma cells in a specific and dose-dependent fashion with an affinity of 32 pM. Competition binding data suggested that murine and human IL-1 compete for a single class of receptor. Antisera were generated in rabbits against both murine and human IL-1. Results of ELISA binding and antisera neutralization assays indicated that there are common antigenic sites between the two IL-1 beta molecules. These domains are of functional importance because they are capable of mediating the neutralization of biologic activity. PMID- 2967327 TI - Enhancement of suppressor T cell activity by injection of anti-IFN-gamma monoclonal antibody. AB - In the present study, the contribution of IFN-gamma to the generation of helper activity in mice was investigated by use of anti-mouse IFN-gamma rat mAB (AN 18.17.24). This mAb was alum precipitated and injected i.p. before or after carrier priming. Results show that spleen cell helper activity is markedly inhibited by anti-IFN-gamma mAb injection. This inhibition is time and dose dependent, and counteracted by IFN-gamma administration. Thus, the anti-IFN-gamma mAb appears to inhibit helper cell activity by neutralization of the IFN-gamma required for the antibody response. Moreover, AN 18.17.24 mAb injection results in increased activation of Lyt-2+ T cells which markedly suppress Th activity. These findings altogether indicate that besides the activation of macrophages and Th, IFN-gamma seems to exert a negative interference in suppressor T lymphocyte circuits and, as a consequence, to inhibit immunosuppression. PMID- 2967328 TI - Islet beta-cytotoxic monoclonal antibody against glycolipids in experimental diabetes induced by low dose streptozotocin and Freund's adjuvant. AB - Diabetes was induced in BALB/c mice by four injections of a subdiabetogenic dose (40 mg/kg) of streptozotocin in combination with CFA. The treatment increased the plasma glucose from 5.8 +/- 0.1 to 22.1 +/- 1.3 mmol/liter (n = 9). The diabetic animals had circulating islet cell surface antibodies (75%), and a monoclonal islet cell surface IgM antibody, K56aF3, generated from one of the diabetic BALB/c mice, mediated C-Dependent cytotoxicity against insulin-producing cells and inhibited glucose-stimulated insulin release from isolated rat islets. Solid phase assay on thin layer chromatograms showed no binding of the K56aF3 antibody to glycolipids prepared from relevant cells. However, testing against a series of glycolipids of various non-pancreatic origins showed a preferential binding to a nine-sugar glycolipid isolated from human erythrocytes carrying an unusual blood group A determinant (type 3). It is suggested that this mAb may be associated with the development of diabetes following a combination of polyclonal activation and non-diabetogenic doses of streptozotocin. PMID- 2967329 TI - Tumor necrosis factor-mediated cytotoxicity involves ADP-ribosylation. AB - The mechanism of TNF-mediated cytotoxicity was studied in several cell lines, including L929 murine fibroblasts. TNF caused a time- and dose-dependent increase of ADP-ribosylation in L929 target cells parallel to cell death. During the course of TNF-mediated cytotoxicity in the presence of actinomycin D, an increase in ADP-ribosylation became apparent between 4 and 6 h after exposure to TNF. Intracellular NAD+ and ATP levels decreased parallel to but not preceding cell death. Two inhibitors of ADP-ribosylation, namely 3-aminobenzamide and nicotinamide, prevented TNF-mediated cytotoxicity. Another target, the human cervical carcinoma cell line ME-180, showed an increase in ADP-ribosylation when treated with TNF, and the cytotoxic action of TNF on this target cell was inhibited by these two inhibitors. In the absence of actinomycin D, treatment of L929 cells with TNF also increased ADP-ribosylation, and the cytotoxic action of TNF was inhibited by nicotinamide. These results indicate that ADP-ribosylation may be involved in the TNF-mediated cytotoxic reaction. PMID- 2967330 TI - IL-4 is an essential factor for the IgE synthesis induced in vitro by human T cell clones and their supernatants. AB - The property of 109 CD4+ T cell clones (TCC) to induce IgE synthesis in vitro in human B cells was compared with their ability to produce IL-2, IL-4, and IFN gamma in their supernatants (SUP) after 24-h stimulation with PHA. A significant positive correlation was found between the property of TCC to induce or enhance spontaneous IgE synthesis and their ability to release IL-4. In contrast, there was an inverse relationship between the IgE helper activity of TCC and their ability to release IFN-gamma, whereas no statistical correlation between the property to induce IgE synthesis and to produce IL-2 was observed. The ability of PHA-SUP from 71 CD4+ TCC to induce IgE synthesis in B cells was also investigated. Twenty-nine SUP (all derived from TCC active on IgE synthesis) induced production of substantial amounts of IgE in target B cells. There was a correlation between the amount of IgE synthesized by B cells in response to these SUP and their IL-4 content. An even higher correlation was found between the IgE synthesis induced by these SUP and the ratio between the amount of IL-4 and IFN gamma present in the same SUP. Like IL-4-containing SUP, rIL-4 also showed the ability to induce IgE production in B cells from both atopic and nonatopic donors. The addition to B cell cultures of anti-IL-4 antibody virtually abolished not only the IgE synthesis induced by rIL-4, but also that stimulated by TCC and their SUP. In contrast, the IgG synthesis induced by TCC SUP was not or only slightly inhibited by the anti-IL-4 antibody. These data indicate that IL-4 is an essential mediator for the IgE synthesis induced in vitro by human TCC and their SUP in the absence of a polyclonal activator, whereas IFN-gamma seems to exert a negative regulatory effect on the production of IgE. PMID- 2967331 TI - Functional heterogeneity among human inducer T cell clones. AB - Analysis of mouse CD4+ inducer T cells at the clonal level has established that a dichotomy among CD4+ T cell clones exists with regard to types of lymphokines secreted. Mouse T cell clones designated Th1 have been shown to secrete IL-2 and IFN-gamma, whereas T cell clones designated Th2 have been shown to produce IL-4 but not IL-2 or IFN-gamma. To determine if such a dichotomy in the helper inducer T cell subset occurred in man, we examined a panel of human CD4+ helper/inducer T cell clones for patterns of lymphokine secretion and for functional activity. We identified human T cell clones which secrete IL-4 but not IL-2 or IFN-gamma, and which appeared to correspond to murine Th2 clones. In marked contrast to murine IL-2 secreting Th1 clones which do not produce IL-4 or IFN-gamma, we observed that some human T cell clones secrete IL-2, and IFN-gamma as well as IL-4. Southern blot analysis indicated that these multi-lymphokine-secreting clones represented the progeny of a single T cell. IL-4 secretion did not always correlated with enhanced ability to induce Ig synthesis. Although one T cell clone which secreted IL-2, IL-4, and IFN-gamma could efficiently induce Ig synthesis, another expressed potent cytolytic and growth inhibitory activity for B cells, and was ineffective or inhibitory in inducing Ig synthesis. These results indicate that although the equivalent of murine Th2 type cells appears to be present in man, the simple division of T cells into a Th1 and Th2 dichotomy may not hold true for human T cells. PMID- 2967333 TI - Giant haemangiopericytoma of abdominal wall. PMID- 2967332 TI - Anti-proliferative effect of IFN-gamma in immune regulation. I. IFN-gamma inhibits the proliferation of Th2 but not Th1 murine helper T lymphocyte clones. AB - A biphasic dose-response curve was observed when the IL-1-dependent HTL clone D10 was exposed to IL-1 plus supernatants from some activated T cell clones but not others. The active component that inhibited proliferation at high concentrations of these supernatants appeared to be IFN-gamma based on the following findings: 1) the biphasic pattern of responsiveness correlated with the presence of IFN gamma in the supernatants; 2) an anti-IFN-gamma mAb augmented the proliferation of D10 cells to these supernatants; 3) rIFN-gamma inhibited profoundly the response of D10 cells stimulated with rIL-1 plus supernatant from activated D10 cells or with rIL-1 plus rIL-4; 4) the response of D10 cells to rIL-1 plus rIL-2 also was inhibited by rIFN-gamma, although to a lesser extent. The proliferation of an additional Th2 clone stimulated with rIL-1 plus rIL-4 or rIL-2 also was inhibited by rIFN-gamma, implicating IFN-gamma as an inhibitory lymphokine for Th2 cells in general. rIFN-gamma did not affect the proliferation of two Th1 clones, nor did it affect the proliferation of an unconventional HTL clone which produces both IL-4 and IFN-gamma and proliferates in response to IL-2 or IL-4 in an IL-1-independent fashion. The proliferation of D10 cells stimulated by Ag or by immobilized anti-CD3 antibody also was blocked by rIFN-gamma, whereas IL-4 production in response to these stimuli was unaffected, indicating that proliferation and not general cell function was specifically inhibited. Collectively, these data implicate IFN-gamma as a suppressive factor for the proliferation of the subset of HTL designated Th2, and suggest that the relative amounts of the various lymphokines present during an immune response may direct which T cell types increase in number. PMID- 2967334 TI - Keratinocyte urokinase-type plasminogen activator is secreted as a single chain precursor. AB - Urokinase-type plasminogen activator (uPA) is produced and secreted by cultured human keratinocytes as a single chain precursor. UPA in keratinocyte conditioned medium is not susceptible to inhibition with diisopropylfluorophosphate (DFP), and it has an apparent molecular weight of 55 kD under both reducing and nonreducing conditions. Cleavage of keratinocyte uPA by plasmin results in the formation of a 96 kD complex comprised of activated uPA and PA inhibitor 2. PA extracted from normal human epidermis is only partially inhibited by DFP, suggesting that precursor uPA is also present in vivo. The synthesis of uPA as a precursor with reduced enzymatic activity as well as decreased affinity for inhibitors is likely to be a mechanism by which normal epidermis regulates plasminogen activation in vivo. PMID- 2967335 TI - Effects of isotretinoin on the neutrophil chemotaxis in cystic acne. PMID- 2967336 TI - Prolonged immunity after late booster doses of hepatitis B vaccine. PMID- 2967338 TI - Lateral segregation of sterol and channel proteins in the mitochondrial outer membrane induced by phospholipase A2: evidence from negative-stain electron microscopy using filipin. AB - The channel protein in the mitochondrial outer membrane of Neurospora crassa aggregates laterally into crystalline arrays by the action of phospholipase A2. When mitochondrial outer membranes are reacted with filipin and examined by negative-stain electron microscopy, filipin-sterol complexes are found everywhere on the membranes except on the crystalline channel arrays. This suggests that the channel-rich membrane domains may have a relatively low content of accessible sterol. It is proposed that in vitro segregation of protein and lipid membrane components by phospholipase A2 may reflect a mechanism by which the endogenous enzyme organizes the native mitochondrial membrane into functional domains. PMID- 2967337 TI - Hepatitis B vaccination in high-risk workers. PMID- 2967339 TI - Ultramicroscopic observations on the pseudointimal coating in implanted dacron vascular grafts. AB - The common use of Dacron arterial prostheses produced enormous improvement in vascular surgery, yet some serious complications such as thrombosis or infection persist, which are connected to the utilization of this material. Light and electron microscopic studies may allow a better understanding of such complications. Scanning electron microscopy and semithick sections were used to examine dacron arterial prostheses removed for failure after long-term implantation. Semithick sections showed periprosthetic inflammatory reaction and the neointimal surface never exhibited a regular endothelial layer (at S.E.M.). PMID- 2967340 TI - Quantitative evaluation of carotid blood flow before and after surgery. AB - Blood flow in 49 patients suffering from stenosing lesions of the supraaortic trunks was measured using an ultrasonic volume flow metre (VFM) before and after operation. Results showed a statistically significant increase in the values of carotid flow, after operation. This non-invasive quantitative determination of blood flow provides the surgeon with often essential information for correct operation and subsequent follow-up. PMID- 2967341 TI - [Laparoscopic diagnosis and evaluation of danazol or gestrinone therapy for endometriosis in sterility]. AB - In forty-six sterile women, endometriosis externa was diagnosed and classified laparoscopically. 21 patients were treated with Gestrinone 5mg-10mg/week and 25 patients were treated with Danazol 300mg-400mg/day for 6 months. The effects of these hormonal treatments were evaluated by second-look laparoscopy according to adhesion severity, number of blueberry spots and chocolate cyst size, as well as dysmenorrhea and other complaints. The results were as follows; 1) Dysmenorrhea was relieved in 60.0% of the Gestrinone-treated group and 45.5% of the Danazol group. 2) Adhesion was weakened or partially separated spontaneously in 66.7% of the Gestrinone group and 63.6% of the Danazol group. 3) Blueberry spots decreased in number or paled in 61.9% of the Gestrinone group and 75.0% of the Danazol group. 4) Chocolate cyst size became smaller in 60.0% of the Gestrinone group and 77.8% of the Danazol group. 5) Peritoneal fluid volume was not decreased after the hormonal treatments but the prostaglandin E2 concentration in peritoneal fluid was decreased (p less than 0.05) after Gestrinone therapy. 6) The patients complained of some side effects, liver function especially was disturbed in 48.0% of the Danazol group and 9.5% of the Gestrinone group. Hoarseness was complained of in 33.3% of the Gestrinone group and 12.0% of the Danazol group. 7) Finally, 23.8% of the Gestrinone group and 28.0% of the Danazol group conceived after the hormonal treatments. PMID- 2967342 TI - The aerobic capacity of locomotory muscles in the tufted duck, Aythya fuligula. AB - The locomotory muscles of the tufted duck, Aythya fuligula (L.), were analysed for mass, aerobic and anaerobic enzyme activities, fibre-type proportions, capillarity, mitochondrial and myoglobin content. The estimated aerobic capacity of the muscles correlated well with the muscles' maximal oxygen uptake both when measured during swimming and when predicted for steady-state flight. The results suggest that exercise performance in birds cannot be predicted purely on the basis of muscle mass (see Butler & Woakes, 1985); the specific enzyme complement of each muscle must also be taken into account. The delivery of oxygen to mitochondria is facilitated by the dense capillarity and high myoglobin content of the muscles. PMID- 2967343 TI - Forgotten but not gone: savings for pictures and words in long-term memory. AB - Five experiments examined the relearning of words, simple line-drawing pictures, and complex photographic pictures after retention intervals of 1 to 10 weeks. For those items that were neither recalled nor recognized, the identical item was relearned better than an unrelated control item, as measured by a recall test following relearning. This relearning advantage in recall held for all three classes of material and extended to the cross-modality case (i.e., picture-word and word-picture) and the same-referent case (i.e., two pictures of the same object). However, recognition tests of relearning failed to detect this same relearning advantage for apparently forgotten items. Taken together, these findings conflict with the existing account of savings. Most fundamental, the classic argument that relearning serves a trace-strengthening function is undetermined by the observed recall-recognition contrast. An alternative explanation of savings is suggested wherein relearning assists retrieval of information, thereby affecting recall in particular. PMID- 2967344 TI - Persistent repetition priming in picture naming and its dissociation from recognition memory. AB - The current experiments investigated the longevity of repetition priming and dissociations between different memory measures. Picture-naming latencies revealed robust repetition priming in four separate studies: Previously named pictures were named faster than new pictures. The magnitude of this naming facilitation was stable across 1 to 6 weeks. The apparent temporal invulnerability of repetition priming was in marked contrast to the decline in episodic recognition memory across 6 weeks, suggesting a dissociation between implicit and explicit memory. Additional evidence of this dissociation was observed within each session: Naming facilitation for repeated pictures occurred regardless of whether those particular pictures were consciously recognized. PMID- 2967345 TI - Frequency encoding of token and type information. AB - Two experiments examined how attention to stimulus attributes affects knowledge of frequency of occurrence. In Experiment 1, orienting tasks were used to direct subjects' attention to either the category membership or the initial letters of words. In Experiment 2, subjects' attention to words, category membership, and initial letters was directed with explicit instructions. The results of these two experiments suggest that attention to specific stimulus attributes may be necessary to initiate the encoding of frequency information. We discuss the implications of these results for claims that the encoding of frequency of occurrence is automatic. PMID- 2967346 TI - Contextually relevant aspects of meaning. AB - A series of six experiments investigated whether inferences about contextually relevant aspects of meaning were encoded into memory during reading. In all the experiments, subjects studied short paragraphs. Then, test sentences were presented that expressed relevant aspects of meaning that had not been explicitly stated in the paragraphs. For example, for a paragraph about searching for the correct color to paint a picture of a tomato, a relevant aspect of meaning would be that tomatoes are red. The test sentences were presented either immediately following the relevant paragraph or after a delay. With immediate testing, it was argued that the facilitation obtained in verification latency could result from processes occurring either when the context was read or when the test sentence was verified. With delayed testing, evidence was found to support the hypothesis that contextually relevant aspects of meaning are incorporated into the memory representation of the paragraph, but such evidence was obtained only when the retrieval environment encouraged the use of newly learned information in the decision process on the test sentence. PMID- 2967347 TI - Formation of complexes between long tail fibres and substructural elements of phage T4D. AB - Complexes of substructural elements of bacteriophage T4 (baseplates, baseplate core complexes) with long tail fibres were obtained for the first time by complementation in vitro. A study of the organization of the complexes was carried out by PAGE, electron microscopy and sedimentation analysis. About 90% of baseplates and baseplate-core complexes were combined with fibres. However, the number of the attached fibres varied from one to six. On the basis of the data obtained, we proposed that the attachment of long tail fibres can occur before the assembly of the whole bacteriophage. PMID- 2967348 TI - Alexithymia and somatization. A Rorschach study of four psychosomatic groups. AB - The construct of alexithymia has been postulated as a predisposing factor in psychosomatic illness. The alexithymia construct has achieved wide currency in psychosomatic research and theorizing despite its doubtful psychometric foundations. Also, the question of between-group variability in alexithymia has not been addressed. In this study we proposed and tested a Rorschach measure of alexithymia on four groups of psychosomatic patients (back pain, gastrointestinal, dermatology, migraine headache). It was hypothesized that psychosomatic groups would be more alexithymic than nonpatients. To examine the question of between-group variability in alexithymia, it was hypothesized that back pain patients would be more alexithymic than other psychosomatic groups. Both hypotheses were supported. Additionally, exploratory comparisons between psychosomatic groups revealed a number of differences between the groups in basic personality processes. The findings show promise for the use of the Rorschach test as an alexithymia measure. Based on the current study, heterogeneous grouping of psychosomatic patients in research designs and treatment programs appear to be a highly questionable procedure. PMID- 2967349 TI - Characterization of [3H]paroxetine binding in rat brain. AB - The binding of the 5-hydroxytryptamine (5-HT, serotonin) uptake inhibitor [3H]paroxetine to rat cortical homogenates has been characterized. The effect of tissue concentration was examined and, with 0.75 mg wet weight tissue/ml in a total volume of 1,600 microliter, the binding was optimized with an apparent dissociation constant (KD) of 0.03-0.05 nM. Competition experiments with 5-HT, citalopram, norzimeldine, and desipramine revealed a high (90%) proportion of displaceable binding that fitted a single-site binding model. Fluoxetine and imipramine revealed, in addition to a high-affinity (nanomolar) site, also a low affinity (micromolar) site representing approximately 10% of the displaceable binding. The specificity of the [3H]paroxetine binding was emphasized by the fact that 5-HT was the only active neurotransmitter bound and that the serotonin S1 and S2 antagonist methysergide was without effect on the binding. Both 5-HT- and fluoxetine-sensitive [3H]paroxetine binding was completely abolished after protease treatment, suggesting that the binding site is of protein nature. Saturation studies with 5-HT (100 microM) sensitive [3H]paroxetine binding were also consistent with a single-site binding model, and the binding was competitively inhibited by 5-HT and imipramine. The number of binding sites (Bmax) for 5-HT-sensitive [3H]paroxetine and [3H]imipramine binding was the same, indicating that the radioligands bind to the same sites. Lesion experiments with p-chloroamphetamine resulted in a binding in frontal and parietal cortices becoming undetectable and a greater than 60% reduction in the striatum and hypothalamus, indicating a selective localization on 5-HT terminals. Together these findings suggest that [3H]paroxetine specifically and selectively labels the substrate recognition site for 5-HT uptake in rat brain. PMID- 2967350 TI - Guanine nucleotide regulation of [125I]beta-endorphin binding to rat brain membranes: monovalent cation requirement. AB - The binding of [125I]beta h-endorphin to rat brain membranes was investigated in the presence of GTP and guanylyl-5'-imidodiphosphate. In contrast to the binding of the mu-selective opioid agonist, [3H][D-Ala2,MePhe4,Glyol5]enkephalin, and the delta-selective opioid agonist, [3H][D-penicillamine2, D penicillamine5]enkephalin, [125I]beta h-endorphin binding was not affected by GTP or guanylyl-5'-imidodiphosphate in a concentration-dependent manner in the absence of cations. However, in the presence of NaCl, the inclusion of either GTP or guanylyl-5'-imidodiphosphate resulted in a concentration-dependent inhibition of [125I]beta h-endorphin binding. This inhibition was significantly greater than the decrease in [125I]beta h-endorphin binding observed in the presence of sodium alone. Although GTP most potently inhibited [125I]beta h-endorphin binding in the presence of sodium, inhibition of [125I]beta h-endorphin binding by GTP was also observed in the presence of the monovalent cations lithium and potassium, but not the divalent cations magnesium, calcium, or manganese. The effect produced by GTP in the presence of NaCl was mimicked by GDP, but not by GMP or other nucleotides. Unlike [125I]beta h-endorphin, the binding of the putative sigma receptor agonist, (+)-[3H]SKF 10,047, was not significantly altered by GTP or guanylyl-5' imidodiphosphate in the absence or presence of sodium. PMID- 2967351 TI - Effect of ethanol administration of striatal D1 and D2 dopamine receptors. AB - Chronic in vivo exposure of rats to ethanol in a complete liquid diet for 14 or 21 days produced a behavioral tolerance to the acute injection of ethanol. After 21 days, but not 14 days, of chronic exposure, there was a significant increase in the maximum density of striatal D1 and D2 dopamine receptors without a change in these receptors' affinities. A 24-h withdrawal from the 21-day exposure did not alter the observed increase in density. Both the level and duration of ethanol exposure appear to be important variables for demonstration of an increase in striatal D1 and D2 dopamine receptors. PMID- 2967352 TI - Brain quinolinic acid in Huntington's disease. AB - Concentrations of the endogenous neurotoxic tryptophan metabolite, quinolinic acid (QA), were measured in postmortem brain tissue obtained from patients with Huntington's disease (HD) and matched controls, using a gas chromatography/mass spectrometry method. There was no significant difference in either the putamen or the frontal cortex between the HD and control groups. These results do not support the hypothesis that increased QA is responsible for neuronal degeneration in HD. PMID- 2967353 TI - Technetium-99m MAG3 kit formulation: preliminary results in normal volunteers and patients with renal failure. AB - Previous studies have shown that [99mTc]mercaptoacetyltriglycine (MAG3) purified by high performance liquid chromatography (HPLC) is a very promising new renal imaging agent which has characteristics very similar to [131I]orthoiodohippurate. An easily prepared kit formulation has been developed and evaluated in ten normal volunteers and three patients on hemodialysis. The average radiochemical purity was 96.6%. There were no adverse reactions. In the volunteers, the relative uptake +/- 1 s.d. was 49.1% +/- 2.6% for the right kidney and 50.9% +/- 2.6% or the left kidney. Urine activity was 71.4% +/- 6.4% of the injected dose at 30 min and 94.4% +/- 2.2% at 180 min. The 60-min plasma clearance was 340.0 +/- 79.0 ml/min and the volume of distribution was 5.15 +/- 1.1I. Approximately 0.5% of the injected dose was present in the gallbladder at 30-60 min postinjection. Gut activity was not present 30-60 min postinjection but reached 1% of the injected dose by 3 hr. In the hemodialysis patients, approximately 1% of the injected dose was present in the gallbladder and 0.5% in the gut at 30-60 min; gut activity increased to approximately 5% at 3 hr. In summary, results using the kit formulation compare favorably to previously published data using the HPLC purified material. Based on these preliminary results, the kit formulation is expected to have widespread clinical utility. PMID- 2967354 TI - Effects of altered physiologic states on clearance and biodistribution of technetium-99m MAG3, iodine-131 OIH, and iodine-125 iothalamate. AB - Technetium-99m mercaptoacetyltriglycine [( 99mTc]MAG3) is a new renal radiopharmaceutical with biologic properties similar to iodine-131 orthoiodohippuric acid [( 131I]OIH). MAG3 may be used as a replacement for [131I]OIH and/or [99mTc]DTPA. For this reason, we compared the effects of several potential adverse clinical conditions on the clearance and biodistribution of MAG3, OIH and a GFR marker. To simulate renal failure, five mice underwent bilateral renal pedical ligation. Twenty-four hours after surgery they were injected with MAG3 and OIH and killed 2 hr postinjection. Compared to sham operated controls, liver activity for MAG3 and OIH increased from 0.2% to 14.1% and 0.1% to 13.9%, respectively, while intestinal activity increased from 1.3% to 8.9% for MAG3 and 0.2% to 7.7% for OIH. Constant infusion studies were performed in rats to evaluate the effects of increased plasma organic acid levels, mannitol diuresis, dehydration, and acid/base imbalance on the clearance of OIH, MAG3, and [125I]iothalamate. No differences were noted between the OIH and MAG3 clearances following diuresis and dehydration and the differences involving acid/base imbalance were minimal. Dehydration depressed the clearance of [125I]iothalamate more than that of OIH or MAG3. Para-aminohippurate (PAH) infusion inhibited the clearance of MAG3 more than OIH supporting proximal tubular transport for MAG3; PAH had no effect on [125I]iothalamate. In summary HPLC purified MAG3 behaved similarly to OIH under adverse physiologic conditions and the data continue to support the use of MAG3 as a potential clinical substitute for OIH. PMID- 2967355 TI - Preparing the company physician to testify at legal proceedings. AB - Company physicians are frequently required to testify as to their findings and opinions in cases where employees' health jeopardizes their work status. The company physician may face conflicting ethical obligations in weighing the physician-patient relationship against the needs of the employer. If the employee seeks outside health care assistance, the company physician may have to testify against other health care professionals. The three most common forums in which the company physician may be asked to submit medical reports and/or testify are arbitration hearings, workers' claims for Social Security disability, and workers' compensation insurance benefits. Company physicians should be aware of the nature of each type of proceeding and they should be prepared to render persuasive expert testimony. PMID- 2967356 TI - The predicament of backache. PMID- 2967357 TI - Allergic eye reaction to photocopier chemicals. PMID- 2967358 TI - Psychiatric disorders and the Workers' Compensation Board. PMID- 2967359 TI - [Experimental model for otitis media with effusion induced by the type III hypersensitivity reaction]. PMID- 2967360 TI - Management of protein S deficiency. PMID- 2967361 TI - Investigation of the distribution of cementum-associated lipopolysaccharides in periodontal disease by scanning electron microscope immunohistochemistry. PMID- 2967362 TI - Progenitor cell kinetics during guided tissue regeneration in experimental periodontal wounds. PMID- 2967363 TI - Regulation of collagen production in fibroblasts cultured from normal and phenytoin-induced hyperplastic human gingiva. PMID- 2967364 TI - Polyclonal induction of IgG antibody forming cells by stimulation with Actinomyces viscosus T14V. PMID- 2967366 TI - The effect of interleukin-1 beta on hyaluronic acid synthesized by adult human gingival fibroblasts in vitro. PMID- 2967365 TI - Indomethacin or flurbiprofen treatment of periodontitis in beagles: effect on crevicular fluid arachidonic acid metabolites compared with effect on alveolar bone loss. PMID- 2967367 TI - Variance components analysis of data from periodontal research. PMID- 2967369 TI - Permeability of rodent junctional epithelium to exogenous protein. PMID- 2967368 TI - Effects of minocycline on fibroblast attachment and spreading. PMID- 2967370 TI - Gingival tissue-produced inhibition of platelet aggregation and the loss of inhibition in streptozotocin-induced diabetic rats. PMID- 2967371 TI - The effects of service dogs on social acknowledgments of people in wheelchairs. AB - Able-bodied people often exhibit behaviors that show them to be socially uncomfortable upon encountering a physically disabled stranger. These behaviors include less eye contact, gaze avoidance, greater personal distance, and briefer social interactions. This study examined whether persons in wheelchairs with service dogs receive more frequent social acknowledgement from able-bodied strangers than people in wheelchairs without dogs receive. Behaviors of passersby were recorded by an observer who followed a person in a wheelchair at a distance of 15 to 30 feet. Observations were made in public areas amid pedestrian traffic, areas such as shopping malls and a college campus. The behaviors of passersby to the person in a wheelchair, with or without a service dog, were recorded, including smiles, conversation, touch, gaze aversion, path avoidance, or no response. Results indicated that both smiles and conversations from passersby increased significantly when the dogs were present. These findings suggest that the benefits of service dogs for their owners extend beyond working tasks to include enhanced opportunities for social exchange. The service dogs substantially reduced the tendency of able-bodied people to ignore or avoid the disabled person. PMID- 2967372 TI - Interventional radiology. Part 2: Vascular procedures. PMID- 2967374 TI - Occurrence and pharmacological significance of metabolic ortho-hydroxylation of 5 and 8-hydroxy-2-(di-n-propylamino)tetralin. AB - Aromatic ortho-hydroxylation in the liver might be one of several possible reasons for the low bioavailabilities of the potent, centrally acting dopaminergic and serotoninergic agonists 5- and 8-hydroxy-2-(di-n propylamino)tetralin, respectively. In vitro and in vivo experiments showed that such an oxidative metabolism did indeed take place. However, the amount of hydroxylated metabolites found in the brain was estimated to represent only 0.3% of the total amount of drug administered. The O-methylation rates of these catechols were also measured in vitro and showed that 5,6-dihydroxy-2-(di-n propylamino)tetralin is a poor substrate for catechol O-methyltransferase (COMT) and that its 7,8-dihydroxy isomer is virtually devoid of substrate activity. No O methylated metabolites were detected in the in vivo samples analyzed. A new synthetic strategy was applied to achieve the isomeric catechols studied. 5 Methoxy- or 8-methoxy-2-(di-n-propylamino)tetralin was lithiated in the ortho position and the metalated species was subsequently quenched in nitrobenzene, yielding the methoxy hydroxy isomers, which were heated in 48% aqueous HBr to achieve the corresponding catechols. PMID- 2967373 TI - Back pain at Greenwich. PMID- 2967375 TI - Synthesis and interaction with uridine phosphorylase of 5'-deoxy-4',5 difluorouridine, a new prodrug of 5-fluorouracil. AB - 5'-Deoxy-4',5-difluorouridine (4'-F-5'-dFUrd) (10) has been synthesized on the basis of the rationale that the labilization of the glycosidic linkage caused by the 4'-fluoro substituent might allow this compound to be a better prodrug form of the anticancer drug 5-fluorouracil (FUra) than is the widely studied fluoropyrimidine 5'-deoxy-5-fluorouridine (5'-dFUrd). The rate of solvolytic hydrolysis of the glycosidic linkage of 4'-F-5'-dFUrd at pH 1 was about 500-fold faster than that of 5'-dFUrd. Since uridine phosphorylase is thought to be the enzyme that causes degradation of 5'-dFUrd in vivo to generate FUra, we compared the substrate interactions of 5'-dFUrd and 4'-F-5'-dUrd with this enzyme. The Vmax for hydrolysis of 4'-F-5'-dFUrd to FUra by uridine phosphorylase was about 5 fold greater than that of 5'-dFUrd, whereas the Km value of 4'-F-5'-dFUrd was 10 fold lower. The combination of these two factors results in 4'-F-5'-dFUrd having a 50-fold higher value of V/K than does 5'-dFUrd. Against L1210 cells in culture, the IC50 value for growth inhibition by 4'-F-5'-dFUrd was 3 X 10(-7) compared to 3 X 10(-6) for 5'-dFUrd. PMID- 2967376 TI - Central dopaminergic and 5-hydroxytryptaminergic effects of C3-methylated derivatives of 8-hydroxy-2-(di-n-propylamino)tetralin. AB - A number of stereochemically well defined C3-methylated derivatives of the potent 5-hydroxytryptamine (5-HT) receptor agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) have been synthesized, and their stereochemical characteristics have been studies by use of NMR spectroscopy, X-ray crystallography, and molecular mechanics calculations. The compounds were tested for activity at central 5-HT and dopamine (DA) receptors, by use of biochemical and behavioral tests in rats. In addition, the ability of the cis- and trans-8 hydroxy-3-methyl-2-(di-n-propylamino)tetralins (15 and 11) to displace [3H]-8-OH DPAT from 5-HT1A binding sites was evaluated. The stereoselectivity of the interaction of 11 and 15 with 5-HT receptors was much greater than that of 8-OH DPAT. Observed rank order of potencies in the 5-HT1A binding assay corresponds to that in the in vivo biochemical assay. PMID- 2967377 TI - A proposed mechanism of action for ALG and ATG in severe acquired aplastic anaemia. AB - In this study we have shown that various batches of anti-lymphocyte globulin (ALG), anti-thymocyte globulin (ATG) and human intravenous immunoglobulin (IV Ig) all contain antibodies with the capacity to block lymphocyte receptors for the Fc region of IgG, i.e., Fc gamma receptors. These antibodies may exert an effect on Fc gamma receptor bearing suppressor T cell function in vivo. Since T cells have been implicated in the pathogenesis of acquired severe aplastic anaemia, it is conceivable that administration of Fc gamma receptor blocking antibodies in the form of ALG, ATG or IV Ig preparations may be important in the treatment of the disease. The level of Fc gamma receptor blocking produced by these IgG preparations was however found to vary from batch to batch and one lymphocyte donor to another. In vivo Fc gamma receptor modulation will therefore only occur when the "correct" batch of IgG is used, thus affording a possible explanation for the variable clinical response of patients to this type of therapy. PMID- 2967379 TI - Intraaortic balloon pump placement through dacron aortofemoral grafts. AB - Occasionally, severe coronary insufficiency necessitates the placement of an intraaortic balloon pump in a patient with a preexisting synthetic aortofemoral bypass graft. Our experience with two such patients suggests that insertion of the intraaortic balloon through a limb of the aortofemoral graft is a safe and effective technique. PMID- 2967380 TI - Graft appendiceal fistulas. AB - We report the fifth case of a graft appendiceal fistula. The four previously reported cases are reviewed. This is the first report of the utility of the tagged red blood cell study in the evaluation of a patient with a graft appendiceal fistula. PMID- 2967378 TI - Relative effectiveness of alternative androgen withdrawal therapies in initiating regression of rat prostate. AB - From a large number of potentially effective androgen withdrawal regimens including bilateral orchiectomy, estrogens, antiandrogens and LHRH agonists alone or in combinations, we compared the ability of 12 different treatment options to mimic the acute results of surgical castration on the rat prostate. Agents were administered s.c. in clinical doses to groups of male rats daily for three days. On day 4 the prostatic tissue was removed and analyzed by conventional methods for whole-tissue and nuclear concentrations of dihydrotestosterone, nuclear androgen receptor and cytoplasmic androgen receptor. Castration-like changes were most pronounced with the synergistic combinations of cyproterone acetate + low dose diethylstilbestrol, and megestrol acetate + low-dose diethylstilbestrol. Comparing the effectiveness of single agents, low-dose diethylstilbestrol was superior to cyproterone acetate, megestrol acetate, flutamide, leuprolide and RU23908. Leuprolide combined with flutamide was superior to leuprolide + cyproterone acetate, leuprolide + cyproterone acetate + low-dose diethylstilbestrol or leuprolide + RU23908 after three days of administration; however, this advantage disappeared when the treatments were extended to seven days. The observations indicate that the most potent androgen withdrawal therapies such as cyproterone acetate + low-dose diethylstilbestrol and megestrol acetate + low-dose diethylstilbestrol at best approximate but do not surpass the early effects of surgical castration. During the same time course, other regimens are characterized by a slower onset of action and a lesser degree of suppression of androgenic mechanisms within the cell. PMID- 2967381 TI - Failed percutaneous transluminal renal angioplasty: experience with lesions requiring operative intervention. PMID- 2967383 TI - A piece of my mind. Walk with me. PMID- 2967382 TI - FDA ponders approaches to curbing adverse effects of drug used against cystic acne. PMID- 2967384 TI - Efficacy of percutaneous transluminal coronary angioplasty in patients with acute myocardial infarction--comparison with intracoronary thrombolysis. AB - We retrospectively compared the efficacy of percutaneous transluminal coronary angioplasty (PTCA) and intracoronary thrombolysis (ICT) in patients with acute myocardial infarction (AMI). The ICT group consisted of 62 consecutive patients who underwent ICT before the introduction of PTCA for AMI and who were considered to be candidates for PTCA based on review of their cine-films. The PTCA group consisted of 92 consecutive patients who underwent PTCA thereafter. The reperfusion rate was significantly higher in the PTCA group than in the ICT group (92.4% vs 71.4%, p less than 0.01) and the residual stenosis was significantly lower in the former. Furthermore, the incidences of reinfarction and post infarction angina were significantly lower in the former than in the latter (3.3% vs 12.9%, p less than 0.05 and 6.5% vs 29.0%, p less than 0.001 respectively). Although the degree of improvement in left ventricular function was influenced by the result of reperfusion, it was not affected by the reperfusion method. Therefore, PTCA did not improve left ventricular function more than ICT unless ICT alone failed to achieve reperfusion. PMID- 2967385 TI - [Effects of isoflurane anesthesia and surgery on plasma concentrations of alpha hANP and other stress hormones in man]. PMID- 2967386 TI - Appearance of a helper activity by mouse serum in cultured lymphoid cells. AB - Manipulations which inhibit the development of non-specific suppressor T cells did not inhibit the development of a helper activity in culture. These manipulations include culturing only those spleen cells nonadherent to Sephadex G 10, CY-treatment of spleen donor mice, or culturing spleen cells in the presence of MS instead of FCS. Further investigation of the helper cell developed in cultures supplemented with MS indicated that both in vivo and in vitro immunizations with SRBC induced the most effective helper activity, and SRBC ghosts or even sonicated SRBC-ghosts were also effective for in vitro immunization. CM containing IL 2 inhibited the development of a helper activity when added on Day 0 but not on Day 4. rIL 2, however, slightly augmented the development of a helper activity even when added on Day 0. The kinetics of anti SRBC PFC response was not changed in the presence of the helper cell and augmentation of the response by the helper cell was seen on all the days tested. Augmentation occurred without further SRBC addition to anti-SRBC PFC generation cultures whereas it did not occur without "intact" responder spleen cells. The helper cell was resistant to X-irradiation and to the treatment with anti-Thy 1.2 plus C' but sensitive to the treatment with anti-Lyt 1.2 plus C'. The activity of nonspecific suppressor T cells dominated over that of the helper cell and the helper activity appeared only when the ratio of suppressor cells to helper cells was reduced to less than 1:4. PMID- 2967387 TI - [A case of honeymoon hepatitis type B during the vaccination against HBV]. PMID- 2967388 TI - [Long-term follow-up study of HBsAb titers in subjects given HB vaccine and timing for additional vaccination]. PMID- 2967389 TI - [Intraplatelet serotonin levels in renal and collagen diseases]. PMID- 2967390 TI - [Experience in manufacturing equipment for the handicapped: welfare of the handicapped observed by an industrial designer]. PMID- 2967391 TI - [Profile: Ms. Chizuko Hagiuda, an actress who delivers messages of life while performing in a wheelchair]. PMID- 2967392 TI - [Interrelations between cardiac rhythm disorders and heart enlargement in patients with exertion-induced stenocardia and pain-free forms of ischemic heart disease]. AB - Data from 24-h ECG monitoring of 137 patients with angina of effort or painless IHD were compared with respective X-ray evidence of heart enlargement. The patients were identified at an epidemiological screening for multifactorial IHD prevention among male residents of Kaunas, aged 40-59 yrs. A correlation was established between the enlargement of the cardiac volume and left compartments and the severity of arrhythmias. PMID- 2967393 TI - [Cardiac rhythm disorders in patients with chronic cor pulmonale]. AB - Continuous ECG monitoring for many hours combined with echocardiography of the right heart, and determination of systolic pulmonary arterial blood pressure (by Burstin's test) and oxygen saturation of arterial blood were conducted in 68 patients with chronic pulmonary heart resulting from chronic nonspecific pulmonary diseases and 18 normal subjects, aged 30-50 years. Three degrees of right-ventricular hypertrophy have been identified on the basis of the electrocardiographic Minnesota code criteria, confirmed echocardiographically. The incidence of cardiac arrhythmias increased dramatically as hypertrophy progressed. Other arrhythmic risk factors are pulmonary arterial hypertension and arterial hypoxemia. PMID- 2967394 TI - [Organizational aspects of proctological services at a hospital]. PMID- 2967395 TI - [Anaerobic gas infection after operations on the abdominal organs]. PMID- 2967397 TI - [Laparoscopic hernioplasty]. PMID- 2967396 TI - [Plastic surgery of posterior wall of the inguinal canal using the pyramidal muscle]. PMID- 2967398 TI - [A case of massive hydropericardium in hypothyroidism simulating cardiomegaly]. PMID- 2967399 TI - [Potentials and results of percutaneous transluminal coronary angioplasty]. PMID- 2967400 TI - Percutaneous transluminal balloon valvuloplasty for congenital pulmonary valvar stenosis: the first case in Singapore and review of the literature. PMID- 2967402 TI - Epidemiological studies of work-related injuries among law enforcement personnel. PMID- 2967401 TI - Occupational health experience in the construction of Phase I of the Mass Rapid Transit System in Singapore. PMID- 2967403 TI - Efficacy of hepatitis B vaccine. PMID- 2967404 TI - Hematoporphyrin derivative uptake and photodynamic therapy in pancreatic carcinoma. AB - Little information is currently available concerning the uptake of porphyrins by pancreatic tumors, or the effect of photodynamic therapy (PDT) on pancreatic cancer. In Syrian golden hamsters (n = 33), the organ distribution of 125I labeled dihematoporphyrin ether (DHE) was studied in a pancreatic cancer model. In the same animal model the effect of PDT was studied using a gold vapor laser for energy delivery 3 hr after the injection of DHE (n = 7). DHE was 2.4 times more concentrated in the pancreatic tumor than in the nontumorous pancreas at 3 hr. Simultaneously there was a considerable accumulation of DHE in the surrounding gastrointestinal tract, causing perforation of the duodenum and jejunum with resultant death in four (57%) animals after PDT. Photodynamic therapy caused extensive tumor necrosis without any obvious effect on the nontumor-bearing pancreas. Damage to the surrounding tissue in the hamster indicates that precautions should be taken if PDT is to be used clinically in pancreatic cancer. Intratumoral injection of DHE may give higher drug concentrations with greater specificity for tumor treatment. PMID- 2967405 TI - Oncogene expression in T-cell lymphoproliferative disorders. AB - We have investigated the expression of oncogenes and other related genes in eleven patients with T-cell lymphoproliferative disorders and ten patients with other hematologic malignancies. The phenotypes of the T-cell disorders were determined using monoclonal antibodies specific for helper or suppressor subsets. RNA preparations were isolated from peripheral blood mononuclear cells and/or lymph node sections, 5'-end labeled with gamma-32P-ATP, and hybridized under stringent conditions to an excess of nitrocellulose-bound specific cloned DNA; autoradiographs were analysed by microdensitometry. Results revealed increased expression of K-ras, v-fps, transferrin receptor, alpha-tubulin and alpha interferon in at least five of six helper T-cell lymphoproliferative disorders, while five of five suppressor T-cell disorders demonstrated levels of hybridization to these clones no higher than background. However, studies of T suppressor disorders demonstrated enhanced levels of beta-interferon-specific RNA in five of five patients, an increase apparent in three of six T-helper chronic lymphoproliferative disorders. These results demonstrate different patterns of gene expression evident in T-helper and T-suppressor abnormalities. PMID- 2967406 TI - Analysis of application possibilities of autoregressive modelling to Doppler blood flow signal spectral analysis. PMID- 2967407 TI - Investigation and improved performance of optical fibre probes in laser Doppler blood flow measurement. PMID- 2967408 TI - [Linear IgA dermatosis and pulmonary tuberculosis in an adult. Apropos of a case]. AB - Linear IgA bullous dermatosis is a rare entity. Only a few cases are reported in association with another systemic disease. A patient with linear IgA bullous dermatosis and pulmonary tuberculosis is presented, and differential diagnosis with dermatitis herpetiformis and bullous pemphigus is made. PMID- 2967409 TI - [Treatment of erythrodermic and pustulous psoriasis with a combination of methotrexate and etretinate]. AB - We present herein our experience concerning the use of etretinate and methotrexate as concurrent therapies in three patients with severe psoriasis. On this regimen, rapid clearing of the psoriatic lesions has been achieved in all patients. The efficacy, safety, indications and dosage schedules for this combination therapy are discussed. No significant side effects have been observed in our patients. However, caution must be exercised in the use of this experimental combination regimen, which should only be used in severe psoriasis. PMID- 2967410 TI - [Pseudo-acne fulminans caused by isotretinoin]. AB - The use of isotretinoin in therapeutical dermatology has proved to be of great benefit in a series of cutaneous processes, especially in cystic and conglobate acne, where it produces excellent results with a dose of 1 mg/kg/daily. However, its use is not without complications, the majority of which are well known, and doubtlessly others will be brought to light. As for a case of pseudoacne fulminans, the possible etiopathogenic mechanisms of this type of reaction, are under discussion. PMID- 2967411 TI - [Palmoplantar keratoderma of the variant type. Study of 3 families]. AB - We have studied 7 patients that belong to three families diagnosed of varians type palmoplantar keratoderma. This type is characterized by the variety of its clinical manifestations, both personal and familial, as also by the great influence of external factors on its evolution. We point out the stable evolution of the disorder and its resistance to general and topical treatment. PMID- 2967412 TI - Dracontiasis in antiquity. PMID- 2967413 TI - Role of central histaminergic mechanism in behavioural depression (swimming despair) in mice. AB - The role of the central histaminergic system in depression was studied by using swimming despair test in mice - a behavioural model of depression. In this test, immobility of mice reflects a state of depression. Intracerebral (ic) injection of histamine (50-200 micrograms) increased significantly the immobility. The H1 receptor blocker mepyramine (2.5-20 mg/kg ip) had no effect while H2-receptor blocker cimetidine (100-200 micrograms ic) caused a significant decrease in immobility. The histamine induced facilitation was blocked completely by cimetidine and antidepressant drugs-imipramine and desipramine, but remained unaffected in mice pretreated with mepyramine or atropine. The H2 agonist impromidine (20-40 micrograms ic) also enhanced significantly, the immobility which was blocked by cimetidine and antidepressant drugs. It has been concluded that central H2-receptors facilitate depression and antidepressant drugs block central H2-receptors. PMID- 2967414 TI - Molecular forms of atrial natriuretic peptides in dog atrium and plasma. AB - The level of immunoreactive ANP (iANP) as determined by radioimmunoassay, exhibits a fairly even distribution throughout the canine atria. However, the maximal concentration was found in the appendages of both the left and right atrium, and the level was significantly higher on the left side in all analyzed localizations. Reverse-phase HPLC of atrial extracts, coupled with RIA, revealed the presence of three iANP fractions. The dominant fraction (65% of iANP) has an estimated molecular weight 16,300, showing that it corresponds to the hormone precursor. By contrast, in the plasma extract, 92% of iANP coelutes with synthetic human ANP99-126, thus confirming the structural identity of dog and human ANP. PMID- 2967415 TI - Linoleic acid requirement of rats fed trans fatty acids. AB - The amount of linoleic acid required to prevent undesirable effects of C18 trans fatty acids was investigated. In a first experiment, six groups of rats were fed diets with a high content of trans fatty acids (20% of energy [en%]), and increasing amounts of linoleic acid (0.4 to 7.1 en%). In a second experiment, four groups of rats were fed diets designed to compare trans fatty acids with saturated and cis-monounsaturated fatty acids of the same chain length at the 2 en% linoleic acid level. After 9-14 weeks, the oxygen uptake, lipid composition and ATP synthesis of heart and liver mitochondria were determined. The phospholipid composition of the mitochondria did not change, but the fatty acid compositions of the two main mitochondrial phospholipids were influenced by the dietary fats. Trans fatty acids were incorporated in all phospholipids investigated. The linoleic acid level in the phospholipids, irrespective of the dietary content of linoleic acid, increased on incorporation of trans fatty acids. The arachidonic acid level had decreased in most phospholipids in animals fed diets containing 2 en% linoleic acid. At higher linoleic acid intakes, the effect of trans fatty acids on the phospholipid arachidonic acid level diminished. However, in heart mitochondrial phosphatidylethanolamine, trans fatty acids significantly increased the arachidonic acid level. Despite these changes in composition, neither the amount of dietary linoleic acid nor the addition of trans fatty acids influenced the mitochondrial function. For rats, a level of 2 en% of linoleic acid is sufficient to prevent undesirable effects of high amounts of dietary C18 trans fatty acids on the mitochondrial function. PMID- 2967416 TI - [Experience in the development of an automated information retrieval system in roentgenoradiology]. AB - The first version of an automated data retrieval system in radiology, radiobiology and oncology has been developed in the Research Institute of Oncology and Medical Radiology of the Ministry of Health, Byelorussian Soviet Socialist Republic. The system is realized on the basis of a packet of applied programs of an automated document processing system, computerized data-bases of the All-Union Scientific and Technical Information Institute and the ES-1022 computer. The system functions in the following modes: 1--selective propagation of information on 194 fixed requests of users; 2--personal search in the dialogue mode; 3--updating of data files. The use of the automated system made it possible to enhance the effectiveness and quality of document search as compared to conventional forms of operation. PMID- 2967417 TI - [The role of solvents in the development of skin changes]. AB - The dermatological tests covered 289 rubber industry workers occupationally exposed to organic solvents and rubber. The control group involved 95 subjects. All the subjects underwent epidermal tests with routine allergens, naephtha and 11 rubber allergens. Altogether, dermatoses have been diagnosed in 32.8% of persons exposed to solvents and rubber, 12.1% of subjects developed dermatitis of hands and forearms, whereas 2.8%--trophic lesions of nails. 6.2% were sensitized to rubber allergens, 13.5%--to chromium compounds, and 3.8%--to cobalt compounds. The number of cases of hand dermatitis and nail lesions was found to increase with age and length of employment. Similarly, the allergy prevalence increased with age and duration of exposure to solvents and rubber. In 40% of dermatitis affected subjects, sensitization to rubber allergens was an aetiological factor, in 60%--hand dermatitis resulted from irritating effects of organic solvents. PMID- 2967418 TI - Sandhoff's disease (type II GM2 gangliosidosis). Clinical, biochemical and ultrastructural study of a case. AB - A case of GM2 gangliosidosis is reported: the diagnosis has been made by clinical findings showing macular cherry-red spot and a progressive neurologic symptomatology with epilepsy, by electron microscopic observation in conjunctival fibroblasts of numerous vacuoles ("zebra bodies") and by absence of total hexosaminidase activity in leukocytes. An abnormal increase of urinary oligosaccharides has also been found. PMID- 2967419 TI - Heritability of variation of plasma cortisol levels. AB - Heritability of the variation of the plasma concentrations of total and unbound cortisol, cortisol binding globulin (CBG), and dehydroepiandrosterone sulfate (DHEA-S) was investigated in 20 monozygotic (MZ) and 20 dizygotic (DZ) male twin pairs. Three plasma samples collected between 8 AM and 9:30 AM were pooled for the assays. Heritability was calculated from the intraclass correlation [2(rMZ - rDZ)]. The mean age, total and unbound cortisol, CBG, and DHEA-S were not significantly different between the MZ and DZ groups of twins. The heritability index for variability of the plasma content of steroids was 45.4% (p less than .05) for total cortisol, 50.6% (P less than .05) for unbound plasma cortisol, 57.8% (P less than .05) for DHEA-S, and 32.4% (P greater than .05) for CBG. The data were analyzed by factor analysis, and heritability estimates were corrected for factors including age, smoking, drinking, exercise, and degree of obesity. These factors did not account for the variation in hormone values in twin pairs. Factor analysis of the three quantitative measurements, cortisol, percent free cortisol, and DHEA-S, provides no evidence for shared factors. The correlation coefficients between age and CBG and total and unbound plasma cortisol concentrations were insignificant. The correlation coefficient between total plasma cortisol levels and CBG was 0.57, which indicates that CBG accounts for 32% of the variation of plasma cortisol concentrations. The results suggest that genetic factors have a decided influence on the variation of the concentration of cortisol of DHEA-S in normal adult men. PMID- 2967420 TI - Immobilized enzymes for thrombolytic therapy. PMID- 2967422 TI - Further evidence for heterogeneity of Fc gamma-receptors on guinea pig splenic B and T lymphocytes: analysis using monoclonal antibodies to two distinct types of Fc gamma-receptor. AB - In our previous paper, we reported that guinea pig splenic lymphocytes expressed two distinct Fc-receptors for homologous IgG (Fc gamma Rs), one monospecific for IgG2 (Fc gamma 2R) and the other bispecific for IgG1 and IgG2 (Fc gamma 1/gamma 2R), when analyzed by EA-rosette assay. These Fc gamma Rs on the cells were further studied by using two monoclonal antibodies toward the Fc gamma Rs on guinea pig peritoneal macrophages (anti-Fc gamma 1/gamma 2R and anti-Fc gamma 2R antibody). The anti-Fc gamma 1/gamma 2R antibody completely inhibited the rosette formation of splenic lymphocytes with IgG1-sensitized sheep erythrocytes [EA(IgG1)]. On the other hand, EA(IgG2)-rosette formation was inhibited partially by anti-Fc gamma 2R but not by anti-Fc gamma 1/gamma 2R antibody. Complete inhibition of the EA (IgG2)-rosette formation was achieved by simultaneous additions of both anti-Fc gamma 2R and anti-Fc gamma 1/gamma 2R antibodies. The binding of IgG2 antibody complexed with ovalbumin to the cells was partially inhibited by either anti-Fc gamma R antibody, and complete inhibition occurred in the presence of both the antibodies, indicating that two types of Fc gamma R, Fc gamma 1/gamma 2R, and Fc gamma 2R, are expressed on the cells. The determination of these Fc gamma Rs on B and T lymphocytes by two-color flow cytometry showed that about 52% of B lymphocytes expressed Fc gamma 1/gamma 2R alone and 32% of the cells expressed both the Fc gamma Rs. On the other hand, about 12% of T lymphocytes was found to express Fc gamma 2R alone and the cells expressing Fc gamma 1/gamma 2R were in the minority (3.8%). T lymphocytes expressing both the Fc gamma Rs were not detected. These results show that guinea pig B lymphocytes bear two types of Fc gamma Rs and are heterogeneous with regard to their Fc gamma Rs and that T lymphocytes express Fc gamma 2R mainly. PMID- 2967421 TI - Bacteriophage lambda DNA packaging: a mutant terminase that is independent of integration host factor. AB - Lambda+ is able to grow in Escherichia coli cells lacking integration host factor (IHF), producing a burst of approximately 25% that produced in IHF+ cells. In vitro, however, we find that the lambda DNA packaging enzyme terminase is strongly dependent on IHF in both cos cleavage reactions and DNA packaging reactions. The cos59 mutation renders lambda dependent on IHF in vivo. The cos59 mutation is a deletion of 3 base pairs at the XmnI site in the cohesive end site (cos) of lambda. Variants of lambda cos59 that were able to grow in the absence of IHF were isolated and found to carry a mutation, called ms1, in the Nu1 gene, which codes for the small subunit of terminase. The Nu1ms1 mutation results in a change of the 40th amino acid of the Nu1 gene product from leucine to phenylalanine. The Nu1ms1 terminase was independent of IHF in packaging reactions in vitro. The results indicate that the mutation either renders terminase: (1) able to utilize some host protein other than IHF, or (2) totally independent of host factors. PMID- 2967423 TI - [Significance of psychological studies of patients with lumbar pain]. PMID- 2967424 TI - [Beta-thromboglobulin levels in the neonatal period]. PMID- 2967425 TI - Prevention of perinatal transmission of hepatitis B virus: prenatal screening of all pregnant women for hepatitis B surface antigen. PMID- 2967426 TI - Lysophospholipid-mediated alterations in the calcium transport systems of skeletal and cardiac muscle sarcoplasmic reticulum. AB - The effects of various lysophospholipids on the calcium transport activity of sarcoplasmic reticulum (SR) from rabbit skeletal and canine cardiac muscles were examined. The lipids decreased calcium transport activity in both membrane types; the effectiveness being in the order lysoPC greater than lsyoPS, lysoPG greater than lysoPE. The maximum inhibition induced by lysoPC, lysoPG and lysoPS was greater than 85% of the normal Ca2+-transport rate. In cardiac SR lysoPE had a maximal inhibition of about 50%. Half maximal inhibition of calcium transport by lysoPC was achieved at 110 nmoles lysoPC/mg SR. At this concentration of lysoPC, the (Ca2+ + Mg2+)-ATPase and Ca2+-uptake activities were inhibited to the same extent (about 60%) in skeletal sarcoplasmic reticulum, while in cardiac sarcoplasmic reticulum, there was less than 20% inhibition of the Ca2+ + Mg2+ ATPase activity. Studies with EGTA-induced passive calcium efflux showed that up to 200 nmoles lysoPC/mg SR did not alter calcium permeability significantly in cardiac sarcoplasmic reticulum. In skeletal muscle membranes the lysophospholipid mediated decrease in calcium uptake correlated well with the increase in passive calcium efflux due to lysophosphatidylcholine. The difference in the lysophospholipid-induced effects on the sarcoplasmic reticulum from the two muscle types probably reflects variations in protein and other membrane components related to the respective calcium transport systems. PMID- 2967428 TI - Characterization of the human monocyte high affinity Fc receptor (hu FcRI). AB - The high affinity Fc receptor (FcRI) of a human monocytic cell line, U937, was further characterized using a previously described murine monoclonal antibody, FcRmAb32. This antibody immunoprecipitated a 70 K cell surface glycoprotein. A solid phase ligand binding assay and a solid phase immunoprecipitation assay were combined to confirm that the 70 K cell surface glycoprotein immunoprecipitated by FcRmAb32 is an IgG binding protein. N-glycanase digestion shows that at least 20% of the relative mobility of the 70 K FcRI glycoprotein is due to N-linked carbohydrate. FcRmAb32 immunoprecipitated a 70 K glycoprotein from biosynthetically labelled U937 cells that co-migrated with the surface iodinated glycoprotein on 2-dimensional gel electrophoresis. A 50 K protein, that is biosynthetically labelled but not accessible to surface iodination, which, bound to control antibodies was also present in FcRmAb32 immunoprecipitates. FcRmAb32 only bound the mature fully glycosylated form of FcRI. The 70 K FcRI was not phosphorylated constitutively nor when U937 cells were stimulated by PMA. PMID- 2967427 TI - Two distinct Fc gamma Rs on guinea-pig polymorphonuclear leukocytes differ from each other in their eliciting activities for O2- generation. AB - When guinea-pig polymorphonuclear leukocytes (PMNs) are stimulated with hen ovalbumin (OA)-complexed IgG antibodies, they generate superoxide anion (O2-). This reaction was found to depend on the IgG isotype used for preparation of the immune complexes; OA-complexed IgG2 antibody (OA-IgG2) induced 3-4 times more intensively O2- generation than OA-complexed IgG1 antibody (OA-IgG1). The O2- generation with OA-IgG1 was almost completely inhibited by a monoclonal antibody to the Fc gamma R binding both IgG1 and IgG2 (Fc gamma 1/gamma 2 R), whereas that with OA-IgG2 was only slightly inhibited. Since guinea-pig PMNs are capable of binding OA-IgG2 not only through Fc gamma 1/gamma 2 R but also through another Fc gamma R which is specific for IgG2 alone (Fc gamma 2 R), the O2- generation with OA-IgG2 may be mainly mediated by Fc gamma 2 R. In addition, cytochalasin B was found to enhance markedly the O2- generation with OA-IgG1, though that with OA IgG2 was only slightly affected. The results so far obtained indicate that Fc gamma 1/gamma 2 R and Fc gamma 2 R differ from each other in their activities for triggering O2- generation, namely activation of the respiratory burst NADPH oxidase. Furthermore, differing from the activation of the NADPH oxidase mediated by Fc gamma 2 R, that by Fc gamma 1/gamma 2 R was shown to be suppressed by some cytochalasin B-inhibitable factor or process though its biochemical nature is unknown. PMID- 2967429 TI - Appearance of acceptor-bound C3b on HLA-DR positive macrophages and on stimulated U937 cells; inhibition of Fc gamma-receptors by the covalently fixed C3 fragments. AB - The appearance and the functional role of acceptor-bound C3b during differentiation of human monocytes into macrophages were studied. Acceptor-bound C3b could be detected by the immune adherence (IA) test parallel to the expression of antigenic determinants specific to mature cells--i.e. on days 4-5 of culture. Consequently, the capacity of these phagocytes to fix C3b covalently via C3b-acceptors (C3bAs) can be considered as one of the signs of their activation/differentiation. All the mature macrophages positive in the IA test were also found to express HLA-DR antigens on their membrane. Using solubilized extracts of stimulated, 35S-cysteine-labelled cells of the human monocytic cell line, U937, we demonstrate that C3 synthesized by these cells can bind to C3bAs of the same cells. Covalently fixed C3 fragments were found to inhibit Fc gamma receptor-mediated ingestion of immune complexes and also antibody-dependent cellular cytotoxicity of monocyte-derived macrophages. PMID- 2967430 TI - [Analgesic intolerance in asthmatic children]. AB - The different clinical presentations of analgesics-intolerance are presented in four asthmatic children. Analgesics-intolerance is rare in children and both sexes are equally affected. The affected children have either a severe mixed asthma and often a chronic sinusitis and nasal polyps, or a chronic urticaria. Often, the first manifestation occurs several years after onset of asthma and is triggered by respiratory infections. Ingestion of most analgesics may cause severe bronchial obstruction, urticaria, angioedema, collaps and rhinitis. The diagnosis can be established by an unequivocal history, or, in uncertain cases, by an inhalation challenge with lysin-acetylsalicylate. The best prophylaxis and therapy is to avoid all responsible drugs. The inhibitory effect of most analgesics on the cyclooxygenase initiates impairments in the metabolism of prostaglandins and leukotrienes, which are suspected to cause the bronchial obstruction in intolerant patients. PMID- 2967431 TI - The myotonic mouse mutant ADR: physiological and histochemical properties of muscle. AB - The muscle physiology and histochemistry of a hereditary neuromuscular syndrome of the mouse, "arrested development of righting response" (ADR), was studied. The speed of single twitches of fast ADR limb muscles was normal up to an age of about 60 days but decreased at later ages. At any age between 10 and 120 days postnatal, fast and slow muscles of the mutant displayed after-contractions of 1 3 (5) seconds duration. These coincided with electrical after-activity of muscle, as demonstrated by electromyography. After-contractions and EMG signals were suppressed by the membrane-stabilizing drug tocainide. These physiological data suggest that ADR is a myotonia. With a few exceptions, limb and trunk muscles of ADR animals showed a uniform oxidative phenotype with a lack of large diameter glycolytic fibers. The histochemical muscle phenotype of the ADR mouse was partially reversed by a long-term treatment with tocainide. PMID- 2967432 TI - Cigarette smoking and levels of adrenal androgens in postmenopausal women. AB - An antiestrogenic effect of cigarette smoking has been suggested, principally on the basis of data on premenopausal women. We examined the relation between cigarette smoking and endogenous sex-hormone levels in a population of 233 white, postmenopausal women 60 to 79 years of age. Current cigarette smokers had significantly higher mean plasma levels of the adrenal androgens dehydroepiandrosterone sulfate and androstenedione than nonsmokers. Mean levels for smokers and nonsmokers were 3.1 mumol per liter (116 micrograms per deciliter) and 2.3 mumol per liter (86 micrograms per deciliter), respectively (P less than 0.001), for dehydroepiandrosterone sulfate, and 27.8 nmol per liter (797 pg per milliliter) and 22.5 nmol per liter (643 pg per milliliter), respectively (P = 0.002), for androstenedione. A dose-response relation was apparent for these hormones; mean plasma levels increased concomitantly with cigarette consumption. The differences in hormone levels remained after adjustment for age and body-mass index. Mean levels of estrone, estradiol, testosterone, and sex-hormone-binding globulin did not differ between smokers and nonsmokers. These results suggest that the possible decreased risk of breast and endometrial cancer associated with cigarette smoking may not be mediated through lower levels of endogenous estrogen, at least in postmenopausal women, and they raise questions about the role of androgens in disease mechanisms in older populations. PMID- 2967433 TI - Aspirin and dipyridamole in the prevention of restenosis after percutaneous transluminal coronary angioplasty. AB - To examine the role of antiplatelet therapy in the prevention of arterial restenosis after percutaneous transluminal coronary angioplasty (PTCA), we conducted a randomized, double-blind, placebo-controlled study in 376 patients. The active treatment consisted of an oral aspirin-dipyridamole combination (330 mg-75 mg) given three times daily, beginning 24 hours before PTCA. Eight hours before PTCA, the oral dipyridamole was replaced with intravenous dipyridamole at a dosage of 10 mg per hour for 24 hours, and oral aspirin was continued. Sixteen hours after PTCA, the initial combination was reinstituted. Treatment was continued in patients with a successfully dilated vessel until follow-up angiography four to seven months after PTCA--or earlier, if symptoms dictated. Of 249 patients who underwent follow-up angiography, 37.7 percent of patients receiving the active drug had restenosis in at least one segment, as compared with 38.6 percent of patients taking placebo (P not significant). The number of stenotic segments was virtually the same in the two groups. Among the 376 randomized patients, there were 16 periprocedural Q-wave myocardial infarctions- 13 in the placebo group and 3 in the active-drug group (6.9 percent vs. 1.6 percent, P = 0.0113). Although the use of this antiplatelet regimen before and after PTCA did not reduce the six-month rate of restenosis after successful coronary angioplasty, it markedly reduced the incidence of transmural myocardial infarction during or soon after PTCA. Thus, the short-term use of antiplatelet agents in relation to PTCA can be recommended. PMID- 2967434 TI - Restenosis after successful coronary angioplasty. Pathophysiology and prevention. PMID- 2967435 TI - The major Fc receptor in blood has a phosphatidylinositol anchor and is deficient in paroxysmal nocturnal haemoglobinuria. AB - Fc receptors on phagocytic cells in the blood mediate binding and clearance of immune complexes, phagocytosis of antibody-opsonized microorganisms, and potently trigger effector functions, including superoxide anion production and antibody dependent cellular cytotoxicity. The Fc receptor type III (Fc gamma R III, CD 16), present in 135,000 sites per cell 1 on neutrophils and accounting for most of FcR in blood, unexpectedly has a phosphatidylinositol glycan (PIG) membrane anchor. Deficiency of Fc gamma R III is observed in paroxysmal nocturnal haemoglobinuria (PNH), an acquired abnormality of haematopoietic cells affecting PIG tail biosynthesis or attachment, and is probably responsible for circulating immune complexes and susceptibility to bacterial infections associated with this disease. Although a growing number of eukaryotic cell-surface proteins with PIG tails are being described, none has thus far been implicated in receptor-mediated endocytosis or in triggering of cell-mediated killing. Our findings on the Fc gamma R III raise the question of how a PIG-tailed protein important in immune complex clearance in vivo and in antibody-dependent killing mediates ligand internalization and cytotoxicity. Together with our results, previous functional studies on Fc gamma R III and Fc gamma R II suggest that these two receptors may cooperate and that the type of membrane anchor is an important mechanism whereby the functional capacity of surface receptors can be regulated. PMID- 2967436 TI - The Fc gamma receptor of natural killer cells is a phospholipid-linked membrane protein. AB - Three types of receptor for the Fc (constant) region of human immunoglobulin G have been described; FcRI, a high-affinity (Ka approximately equal to 10(8) M-1) receptor expressed on monocytes; FcRII (CD32), a low-affinity (Ka approximately equal to 10(6) M-1) receptor expressed on B cells, granulocytes, macrophages and platelets; and FcRIII (CD16, FcRIo), a low-affinity receptor expressed on macrophages, neutrophils, eosinophils, natural killer cells and a subset of T cells believed to comprise the suppressor cells. Anti-CD16 antibodies block natural killer-cell mediated antibody dependent cellular cytotoxicity (ADCC). Binding of aggregated IgG to CD16 on natural killer cells leads to the expression of lymphocyte activation antigens, mediator release, morphological changes and lytic activity. We report here the isolation of a complementary DNA clone encoding CD16 determinants which gave rise to IgG binding of the expected affinity and subtype specificity in COS cells, and which proved to encode a phospholipid anchored protein. A single messenger RNA transcript was found in all positive RNA samples, and N-glycanase treatment showed the form found in COS cells was identical to the form present on peripheral blood mononuclear cells (PBMCs). We also show that CD16 is most closely related to the alpha-form of the murine IgG 2b/1 receptor and propose that extracellular contacts mediate the signal initiated by IgG binding. PMID- 2967438 TI - North Carolina Workers' Compensation. PMID- 2967437 TI - The PI-linked receptor FcRIII is released on stimulation of neutrophils. AB - Human phagocytic cells express receptors for the constant (Fc) region of immunoglobulin G. Neutrophils carry Fc receptor II (FcRII; CDw32) and FcRIII (CD16) which both bind IgG-containing immune complexes, leading to phagocytosis of the complex and activation of the neutrophil. We find that patients with paroxysmal nocturnal haemoglobinuria (PNH) have only about 10% of the normal levels of FcRIII on their neutrophils, whereas the expression of FcRII is unaffected. We show that FcRIII is a phosphatidyl inositol (PI)-anchored protein in neutrophils. Analysis of FcRIII expression in cells of PNH patients, known to be deficient in PI-linked proteins, suggests FcRIII is not PI-linked in monocytes. We find that the synthesis of FcRIII in neutrophils from PNH patients appears normal, indicating that the defect lies in the PI linkage. This lipid linkage of the receptor on neutrophils suggests that its release may be important for its function, and indeed FcRIII release was observed on stimulation of neutrophils by an inflammatory bacterial peptide (f-Met-Leu-Phe), suggesting a role for FcRIII shedding in inflammatory reactions. Activation of the PNH neutrophils with IgG-coated latex beads appeared normal (although binding of dimer IgG complexes was reduced), indicating that FcRII, rather than FcRIII, is involved in neutrophil stimulation. PMID- 2967439 TI - [Relation between low backache and posture habits in women in The Netherlands and Japan]. PMID- 2967440 TI - Protective effect of 13-cis-retinoic acid on hematoporphyrin derivative-induced cell photosensitivity in vitro. AB - The influence of 13-cis-retinoic acid on the light-caused injury of hematoporphyrin-pretreated nontransformed 10 T1/2 fibroblasts and N2a neuroblastoma cells was investigated. Retinoid reduced the percentage of damaged cells as assessed by the plating efficiency assay. N2a neuroblastoma cells were found to be better protected than 10 T1/2 cells by 13-cis-retinoic acid on hematoporphyrin light-induced injury. PMID- 2967441 TI - Retrograde amnesia in patients with Alzheimer's disease or Huntington's disease. AB - Retrograde amnesia (RA) was studied in patients with Huntington's disease (HD) or Alzheimer's disease (AD) using an updated version of the remote memory battery originally developed by Albert, Butters and Levin. Regardless of whether remote memory was measured by unaided recall or cued recall, HD patients exhibited deficits that were equally severe across decades. RA was more severe in AD than in HD patients and the AD patients recalled significantly more items from the 1940s and 50s than from the 60s, 70s or 80s. The AD patients also displayed dysnomia, while the HD patients did not. Naming difficulties appeared to contribute to the poor overall performance of the AD patients, but did not account for the temporal gradient of their RA. These findings, like recent reports focusing on these patients' ability to learn new information and to search semantic memory, indicate that the processes underlying AD and HD patients' memory failures are distinct. PMID- 2967442 TI - The mdx mouse skeletal muscle myopathy: I. A histological, morphometric and biochemical investigation. AB - Skeletal muscle has been examined in a colony of the mdx strain of myopathic mice. Sixty-five mice from 22 to 303 days of age, showed extensive and recurrent areas of necrosis and regeneration of muscle fibres, often accompanied by active cellular infiltration. Morphometry of the soleus muscle revealed an abnormal proportion of small and large muscle fibres; over half of the muscle fibres contained 'central' (non-peripheral) nuclei. No histochemical muscle fibre-type grouping was detected. Serum activities of muscle-derived enzymes were greatly elevated in all animals and probably reflect enzyme leakage from damaged muscle fibres. Histological evidence of a cardiomyopathy was found in 13 mice. The mdx myopathy thus shows features seen in Duchenne muscular dystrophy. Mdx differs from Duchenne dystrophy principally in that it exhibits a greater degree of compensatory muscle regeneration and an absence of fibro-fatty replacement of muscle fibres. PMID- 2967443 TI - Sequential formation of beta-endorphin-related peptides in porcine pituitary. AB - Lipotropin and peptides related to beta-endorphin were extracted from the anterior pituitary and the pars intermedia of porcine pituitary and were resolved by gel exclusion and ion exchange chromatography. Possible heterogeneity in the structure of the lipotropin was investigated by identifying the C-terminal fragment released by limited proteolysis with trypsin; the cleavage was restricted to the carboxyl group of arginine residues by employing citraconylation to protect the epsilon-NH2 groups of lysine. The lipotropin obtained from both regions of the pituitary gave rise to the same C-terminal peptide which contained the 31-residue sequence of beta-endorphin; none of the 26 and 27-residue forms was detected. In contrast, the beta-endorphin-related peptides that were isolated directly from the pars intermedia exhibited a high degree of C-terminal proteolysis: they were present principally as the 26- and 27 residue peptides. The results demonstrate that lipotropin differs from beta endorphin in that it occurs exclusively in the form that contains the full C terminal sequence. It is concluded that during biosynthesis lipotropin undergoes conversion to beta-endorphin before proteolysis takes place at the C-terminus. The processing reactions that convert lipotropin to beta-endorphin 1-31 and beta endorphin 1-31 to beta-endorphin 1-27 are thus ordered and not competitive. The results also indicate that glycylglutamine, the bioactive C-terminal dipeptide of lipotropin, is formed from beta-endorphin and not from lipotropin. PMID- 2967445 TI - [Skin-sandwich free-graft in the treatment of recurrent inguinal hernias]. PMID- 2967444 TI - Presence of beta-hexosaminidase A alpha-chain mRNA in two different variants of GM2-gangliosidosis. AB - Tay-Sachs disease displays a variety of forms on the clinical and biochemical level. On the molecular level it has been shown, that poly (A)+ RNA preparations from fibroblasts of patients with classical Tay-Sachs disease lack detectable alpha-chain message when analyzed by Northern blotting with complementary DNA encoding the alpha-chain of human beta-hexosaminidase A. In this report the p beta H alpha-5 clone was used to investigate whether patients with two different variants of Tay-Sachs disease also lack the alpha-chain message. On the basis of RNA hybridization analyses, we could show that our patients which synthesize an altered alpha-chain, as judged by testing enzyme activity and substrate specificity, have the 2.1 kb mRNA which is also seen in healthy control patients. PMID- 2967446 TI - [A rare congenital malformation of the bile ducts: presence of hepato-cholecystic ducts with agenesis of the common bile duct]. PMID- 2967447 TI - Back injury and low back pain. PMID- 2967448 TI - Comparison of a fenoterol/ipratropium combination with salbutamol from metered dose inhalers in subjects with chronic partially reversible airways obstruction. AB - We have compared the acute bronchodilator response following salbutamol 200 micrograms and fenoterol 200 micrograms with ipratropium 80 micrograms, from metered dose inhalers in ten subjects with severe partially reversible airflow obstruction. Treatment with the combination resulted in a similar degree of bronchodilation but with a significantly longer duration of action. PMID- 2967449 TI - Nebulised bronchodilator therapy in acute severe asthma. PMID- 2967450 TI - Attitudes towards cardiopulmonary resuscitation in a group of people with severe disability. AB - In a prospective survey of fifty admissions to a young disabled unit, more than 75% of people with a severe disability expressed a wish to have cardiopulmonary resuscitation should they suddenly collapse. The opportunity to make an informed decision was positively received by all patients, and the information gained is of value in guiding resuscitation decisions. PMID- 2967451 TI - Antibody response to half dose hepatitis B vaccine. PMID- 2967452 TI - Exfoliative dermatitis: presenting sign of fallopian tube carcinoma. AB - A woman presented with severe exfoliative dermatitis and a pelvic mass subsequently found to be fallopian tube carcinoma. After resection of the tumor and four courses of cisplatin, doxorubicin, and cyclophosphamide, the skin condition cleared. PMID- 2967453 TI - Spontaneous resolution of fetal cystic hygroma in Down's syndrome. AB - We describe a case of spontaneous resolution of a cystic hygroma between 14-16 weeks' gestation in a fetus with trisomy 21 who, at termination at 19 weeks, revealed only mild webbing of the neck. Redundant nuchal skin folds are among the most common features of trisomy 21. Our case supports the hypothesis that this redundant skin of the fetal neck represents early cystic hygromas, which resolve in utero before 16 weeks' gestation. PMID- 2967454 TI - Idiopathic hydrops fetalis successfully treated in utero. AB - Using ultrasound, nonimmunologic hydrops fetalis can be diagnosed antenatally, but the antenatal management remains debatable. To treat a fetus with idiopathic hydrops fetalis, we injected albumin into the fetal abdomen in utero. After this treatment, the hourly fetal urine production rate increased significantly, and edema and ascites disappeared subsequently at two weeks and three weeks, respectively. Edema and ascites were absent at birth. The child remains well at 18 months of age. PMID- 2967455 TI - Differential transformation of primary human embryo retinal cells by adenovirus E1 regions and combinations of E1A + ras. AB - The efficiency of transformation of primary human embryo retinal (HER) cells by the adenovirus E1 region (E1A + E1B) depended on the virus serotype whereas transformation by E1A alone was a rare event regardless of serotype. Activated human c-Ha-ras and N-ras genes co-operated differentially with different E1As for HER transformation but were ineffective without E1A. Ras + E1A co-transformants containing Ad 12 E1A established directly from foci, in contrast to those containing Ad 2 or Ad 5 E1A. A spectrum of activated ras gene expression was found in stable co-transformants with mRNA and protein levels being lower in Ad 12 E1A + N-ras than Ad 2 E1A + N-ras cell lines. Down regulation of E1A transcription in the absence of E1B was found in Ad 2 E1A + Ha-ras transformants only but E1A protein levels were similar to those in Ad 2 E1A + N-ras or Ad 5 E1A + E1B cell lines. HER cell transformants which contained Ad 12 E1A were more tumourigenic than those which contained the Ad 2 or Ad 5 E1A. This unique transformation system shows that stable malignant transformation of primary human cells in vitro is a complex process requiring the combined activities of two or more types of genes. PMID- 2967456 TI - A comparative study of infections with rodent isolates of Giardia duodenalis in inbred strains of rats and mice and in hypothymic nude rats. AB - Giardia isolated from mice and rats have been identified as Giardia duodenalis by morphological criteria. No differences in the electrophoretic mobilities of 28 enzymes were detected between the mouse and rat isolates or between these isolates and human and cat isolates. Infections with both rodent isolates have been studied in several strains of inbred rats. No significant differences were detected between the rat strains, with the mouse isolate producing a self limiting infection and the rat isolate a chronic infection. After the primary infection was eradicated with metronidazole, all strains of rats were resistant to reinfection with the homologous isolate. Both isolates produced chronic infections in hypothymic nude rats. BALB/c mice were found to be relatively resistant to primary infection with either isolate but C3H mice became infected chronically with the mouse isolate and experienced a prolonged infection with the rat isolate. These findings resemble those observed with infections with G. muris in the same strains of mice (Roberts-Thomson & Mitchell 1978). It is suggested that infections with G. duodenalis in rats may offer a model for giardiasis that is based on organisms related closely to G. lamblia. Comparative studies using the two rodent isolates may lead to a better understanding of how the parasite establishes as a chronic infection and which antigens induce protective immune responses. PMID- 2967457 TI - Biliary antibody response in rats infected with rodent Giardia duodenalis isolates. AB - Using a sensitive ELISA, specific serum and bile anti-Giardia IgM and IgA responses were studied in rats infected with two strains of Giardia duodenalis: a rat isolate which produces a chronic infection and a mouse isolate which produces a self-limiting infection. Paired samples of serum and bile were collected from groups of DA (RT1avl) rats at various times during primary and secondary infections. Antibody responses to both organisms were similar. Only IgA anti Giardia antibodies were detected in bile whereas both IgM and IgA antibodies were detected in serum. Biliary IgA antibody titres increased throughout the course of the primary infection and remained at high levels for at least 10 weeks. Biliary IgA titres increased 16-fold during the secondary infection with both isolates. Serum IgA anti-Giardia titres also increased but more slowly than the titres in bile. Serum IgM antibody responses were observed against both organisms during the primary and secondary infections. Trophozoites harvested from the intestinal lumen during primary infections were examined for surface-bound IgA by immunofluorescence microscopy. IgA was detected on 3% of trophozoites on day 7 after infection but on over 70% of trophozoites by the 10th day. The data demonstrate the occurrence of a secretory IgA immune response in rats infected with both G. duodenalis isolates, some of which is directed against surface antigens of the trophozoites. PMID- 2967458 TI - [The mixed lympho-epidermal culture: a predictive test of the graft-versus-host reaction in receivers of bone marrow grafts]. AB - Mixed lymphocyte reactions (MLR) and mixed epidermal cell-lymphocyte reactions (MECLR) were performed concurrently before grafting in patients scheduled to receive a bone marrow graft from an HLA-identical sibling for an hematological malignancy. For each donor-recipient pair, the ratio of the counts per minute (cpm) for MECLR to the cpm for MLR was calculated. This ratio was significantly higher in the group of patients who subsequently developed grade II to IV graft vs-host (GVH) disease as compared to the group who had grade O-I GVH. Statistical analysis showed that this parameter was the most significant risk factor for acute GVH, as it was the leading factor selected by stepwise discriminant analysis. The other significant risk factors for acute GVH were previous pregnancies in female donors and chronic myeloid leukemia in recipients. Thus, these parameters allow more accurate evaluation of the risk for GVH in individual recipients before grafting and therefore of the indications for T-cell depletion. PMID- 2967459 TI - Viral hepatitis. AB - Recent research has led to a greater understanding of the mechanisms and management of the various forms of viral hepatitis. The clinician can rapidly arrive at a precise diagnosis using serologic markers to complement epidemiologic data. In addition, effective immunoprophylaxis is possible; thus, disease spread can be minimized. PMID- 2967460 TI - Relative contribution of ribcage and abdomen during augmented breaths in infants. AB - Changes in the relative contribution of ribcage and abdomen during augmented breaths were assessed in two groups of infants (less than 2 weeks and 4-12 weeks old). In both groups ribcage and abdominal movements increased during augmented breaths in quiet sleep. In the older infants the relative increase was greater for the ribcage (P less than 0.05) when expressed either as a ribcage to abdominal movement ratio or as a percent of the respective movements during control breaths. In contrast, no significant change of the relative contribution was observed in the younger age group. During active sleep paradoxical movement of the ribcage, observed during tidal breaths, continued during the first phase of the augmented breath; however, during the second phase of inspiration, the ribcage moved outward in all but one infant. Our results show that marked increases in tidal volume are associated with greater increases in ribcage contribution in infants beyond the neonatal period. This may reflect differences in recruitment patterns of intercostal muscles during development. PMID- 2967461 TI - [Non-hematological side effects of cytostatic drugs used in children]. PMID- 2967462 TI - [Joint work experience of a department of pediatrics with public health organs and institutions in medical care for a pediatric population]. PMID- 2967463 TI - [Beta-N-acetylhexosaminidase activity of the blood serum of children with different forms of central nervous system pathology]. PMID- 2967464 TI - Isolation of the gene coding for the alpha subunit of the human high affinity IgE receptor. PMID- 2967465 TI - The integration host factor of Escherichia coli binds to multiple sites at plasmid R6K gamma origin and is essential for replication. AB - Examination of the effect of the himA and himD mutants of E. coli on the maintenance of plasmid R6K has revealed that the gamma origin-containing replicons cannot be established in any of the mutants deficient in the production of E. coli Integration Host Factor (IHF). Contrary, the R6K derivatives containing other origins of the plasmid (alpha and/or beta) replicate in a host lacking functional IHF protein. We show that IHF protein binds specifically to a segment of the replication region which is essential for the activity of all three R6K origins. Mapping the IHF binding sequence with neocarzinostatin showed that the protein protects three segments of the origin: two strong binding sites reside within an AT-rich block, while the third, considerably weaker site is separated from the other two by a cluster of the seven 22 bp direct repeats. These seven repeats have been shown previously to bind the R6K-encoded initiator protein pi. We also demonstrate that the establishment of pi-origin complexes prior to IHF addition prevents the binding of the IHF protein to the gamma origin. The binding sequences of IHF and pi proteins do not overlap, therefore, we propose that the binding of pi protein alters the structure of the DNA and thereby prevents the subsequent binding of IHF protein. PMID- 2967466 TI - [Prevention of the horizontal transmission of hepatitis B in children]. AB - The active immunisation by means of vaccination is the best strategy against the B hepatitis virus. Hevac B vaccine was administered to sixty one children (30 boys and 31 girls) aged 1 month 14 years co-inhabitants with chronic carries of virus. After administering three dosages, seroconversion, giving protecting titres (greater than or equal to mU/ml) was obtained in 57 children. Levels in three boys and one girl were not protective. Retarded immunity examination showed alterations of T-Lymphocyte subpopulations only in the girl. The four non responders received a supplementary vaccination a month later, beside the booster dose given one year later (T12). Only two boys had no seroconversion. In spite of the four vaccination four children had antibody titers below 100 mU/ml. PMID- 2967467 TI - [Laparoscopy in oncological pediatrics]. AB - Laparoscopy permits direct inspection of the peritoneal cavity. Its role is very important in oncology both in the adult and in the pediatric patient. From 1973 to 1985, 147 laparoscopies were performed on 137 patients, aged 2-16; indications were: definition of the nature of neoplastic disease in 8 cases, staging of known neoplasia in 132, restaging in 7. On Hodgkin's disease, splenic involvement in 8/113 cases was revealed by laparoscopy with biopsy. Histological examination was positive in 4 out of 22 cases of ovarian tumors. Morbidity was 1.2%. Although a laparotomy performed after laparoscopy has shown some false negatives, in most cases the clinical and instrumental follow-up has proved reliability of laparoscopic examination. PMID- 2967468 TI - 5 alpha-reductase activity in epithelium and stroma of prostates from intact and castrated dogs treated with androstenedione, the aromatase inhibitor 1-methyl-1,4 androstadiene-3,17-dione, and cyproterone acetate. AB - In addition to the histology of epithelial and stromal elements of prostates from intact dogs (group 0) and castrated dogs (group I), the latter of which were treated with androstenedione (group II), androstenedione plus the aromatase inhibitor 1-methyl-1,4-androstadiene-3,17-dione (group III), or androstenedione plus aromatase inhibitor and cyproterone acetate (group IV) (Habenicht and El Etreby: The Prostate 11:133-143, 1987) it was of interest to study the influence of such in vivo treatment on the prostatic 5 alpha-reductase, which is responsible for the cellular conversion of testosterone to 5 alpha dihydrotestosterone. Michaelis constants (KM) and maximal activities (Vmax) of 5 alpha-reductase were determined under optimized incubation conditions in mechanically separated epithelium and stroma. The metabolites were separated by high-performance liquid chromatography and determined radiometrically. The main results were: 1) The mean KM (nM +/- SEM) was significantly (P less than .001) higher in epithelium (892 +/- 132) than stroma (70 +/- 11). The same was true concerning the Vmax (pmol.mg protein-1.h-1 +/- SEM) in epithelium (54.6 +/- 5.8) as compared to stroma (13.0 +/- 2.0). 2) No specific in vivo or in vitro effect of the aromatase inhibitor on the KM and Vmax data was found. 3) In prostates of intact dogs and dogs of group II the proportion of epithelial 5 alpha-reductase exceeded distinctly that of stromal 5 alpha-reductase. 4) In groups I, III, and IV the proportion of epithelial 5 alpha-reductase was rather low. These data were discussed in the light of the histological findings. PMID- 2967469 TI - [Mechanism of antigen presentation by B cells to helper T cells]. PMID- 2967470 TI - [New methods of locating and treating malignant neoplasms by using a hematoporphyrin derivative and photoradiation]. PMID- 2967471 TI - Good backs and bad backs: a general practice survey. PMID- 2967472 TI - Pharmacokinetics of etretin and etretinate during long-term treatment of psoriasis patients. AB - The aromatic retinoic acid derivative etretin has recently been introduced in the treatment of severe psoriasis and other dyskeratoses. Hitherto, the use of the carboxylic acid ester analogue, etretinate, has been hampered by an extremely long elimination half-life of up to 120 days for this drug. Seven patients of either sex from whom we recently reported single-dose pharmacokinetics have been studied after 1 and 3 months multiple dose administration of the drugs. Four were given etretin and three etretinate. Etretin, both as drug and as metabolite, was absorbed faster than etretinate as judged from t-lag, tm and t 1/2 ka. Etretin as drug was eliminated faster than the metabolite etretin, t 1/2 beta 2.39 +/- 1.16 days compared to 6.51 +/- 2.06 days. In patients receiving etretinate the terminal disposition or elimination half-lives for cisetretin (t 1/2 lambda 3 15.9 +/- 9.9 days) were longer than for the metabolite etretin and exhibit a pronounced interindividual variation from 4.25 to 22.8 days. Similarly, cis etretin accumulated very marked in comparison to the metabolite etretin of the drug etretinate. Assuming 40% systemic availability for both drugs, the central compartment of distribution constituted about 12-32% in case of etretin and about 0.8-3.6% in case of etretinate of the calculated apparent total volume of distribution at steady state, which showed mean values of 3.5 and 39.6 1.kg.-1, respectively, presumably reflecting the higher lipophilic nature of the latter compound. PMID- 2967473 TI - First trimester prenatal diagnosis of Sandhoff's disease. AB - Chorionic villus sampling was performed on two patients with a previous family history of Sandhoff's disease. Total beta-hexosaminidase (Hex) activity in case 1 was within the normal range (case 1: 6365 mumol/h/g protein; control range: 3227 24 495 mumol/h/g protein). The beta-hexosaminidase isoenzyme pattern was found to be normal. These results were confirmed on cultured amniotic fluid cells. In case 2, the total Hex activity was 672 mumol/h/g protein, i.e., 7 per cent of the control mean (10,085 mumol/h/g protein), and chromatography demonstrated that more than 50 per cent of this activity was due to the abnormal isoenzyme beta hexosaminidase S (Hex S). The fetus was predicted to be affected by Sandhoff's disease and this was confirmed on fetal tissues after termination of pregnancy. This study demonstrates that a fetus affected by Sandhoff's disease can be reliably diagnosed during the first trimester of pregnancy. PMID- 2967474 TI - [Detection of dislocation of the hip in the newborn infant and infant. Contribution of ultrasonics]. PMID- 2967475 TI - [What is a severe epilepsy?]. PMID- 2967476 TI - [Purulent pleurisy caused by anaerobic bacteria. A retrospective study of 19 cases]. AB - A retrospective clinical, biological, radiological and evolutive study of 19 cases of pleural empyema caused by anaerobic organisms diagnosed between 1980 and 1986 was carried out. These 19 cases accounted for 30.6 p. 100 of all cases of pleural empyema diagnosed during the same period. A local or general contributory factor was found in all patients; false passage, gastrointestinal pathology and buccal or dental diseases were the most frequent aetiological circumstances. The clinical picture was rather torpid, with a body temperature below 38 degrees C in 42 p. 100 of the cases, which delayed the diagnosis: the mean time interval between onset and diagnosis was 20 days. In nearly one half of the cases, blood stained expectoration was present and air-fluid levels were visualized at standard radiography and computerized tomography, which is the best exploratory method to evaluate the size and appearance of the pleural lesion, to guide percutaneous drainage and later to assess possible sequelae. The predominant anaerobic flora consisted of Gram-positive cocci and Bacteroides spp; in 70 p. 100 of the cases the anaerobic organism(s) was (were) sensitive to penicillin G. The course of the disease was favourable in all patients. Surgery was performed in 3 of the 19 patients on account of chest wall gangrene due to Clostridium perfringens in 1 case and of the presence of multiple fluid pockets making drainage ineffective in 2 cases. PMID- 2967477 TI - [Comparative serum pharmacokinetics of prednisone and prednisolone methylsulfobenzoate after oral administration]. AB - Pharmacokinetic values of prednisone and prednisolone were measured in the serum of 6 healthy volunteers after oral administration of either prednisolone methylsulfobenzoate 30 mg or prednisone 30 mg. Peak serum concentrations of prednisolone obtained after dosing with prednisone occurred earlier and were higher (473 +/- 106 ng/ml) than those obtained after dosing with prednisolone methylsulfobenzoate (232 +/- 70 ng/ml; P less than 0.01). Similarly, areas under the 0-8 h concentration curves were significantly greater after dosing with prednisone than after dosing with prednisolone methylsulfobenzoate (prednisolone: P less than 0.001; prednisone: P less than 0.02). The differences may be due to prednisolone methylsulfobenzoate not being absorbed as well as prednisone. These kinetic data may warrant a reappraisal of the therapeutic equivalence of the two drugs taken for granted in France. PMID- 2967479 TI - [Aerosol pulmonary scintigraphy. A new test in the diagnostic strategy of pneumocystis carinii pneumonia in patients with acquired immunodeficiency syndrome]. AB - In eleven patients with AIDS and suspected Pneumocystis carinii pneumonia we measured the pulmonary clearance and half-life of aerosolized 99m Tc-DTPA (diethylenetriamine pentaacetate). We correlated the half-life with gallium scans and bronchoalveolar lavages. In all nine patients with Pneumocystis, this half life was shorter than normal value, documenting a significant increase of lung epithelial permeability (p less than 0.001). Six of these 9 patients had abnormal gallium scans. However, in two patients with normal chest X-rays, PaO2 and gallium scans, only the reduced half-life of 99mTc-DTPA evidenced acute lung disease. After treatment, the results of the DTPA half-life correlated with the presence or absence of Pneumocystis. This method is, therefore, a sensitive initial diagnostic test in patients with suspected Pneumocystis pneumonia. A short half-life should prompt bronchoalveolar lavage even when PaO2 and/or chest X-rays and gallium scans are normal. PMID- 2967478 TI - [AIDS and HIV infection in Guadeloupe]. AB - By December 31, 1986, 38 cases of AIDS had been notified in Guadeloupe (French West Indies). Transmission of the disease was predominantly heterosexual; 79 p. 100 of the patients were devoid of risk factor. Sex ratio was 2.2, but the disease, which initially was mostly observed in men, seems to be developing rapidly in women. Fearing an increase in infantile mortality due to AIDS, the authors recommend routine screening of pregnant women for anti-HIV antibody. PMID- 2967480 TI - [Antibiotherapy in gangrene of the perineum]. AB - Fifty cases of extensive and necrotizing perineal cellulitis ("perineal gangrene") are reported. The disease, remarkable for its many anatomical and bacteriological varieties, is regarded as a therapeutic requiring multidisciplinary management. Bacteriological samples were polymicrobial in 40 cases. Streptococcus D and aerobic Gram-negative bacilli were isolated in 21 patients. The initial antibiotic treatment had to be modified on ten occasions in 36 patients who were given penicillin G in high doses and on five occasions in 14 patients who received a beta-lactam antibiotic active against anaerobes. Nineteen patients (38 p. 100) died; death was directly related to the sepsis in 16 cases. The death rate was the same in the two treatment groups (11/36 and 5/14) and similar to that found in the literature. The authors advocate an ureidopenicillin, fosfomycin and imidazole combination as a substitute for the conventional penicillin G, aminoglycoside, imidazole combination, since the former is active against Streptococcus D and has better tissue penetration. PMID- 2967481 TI - [The centrimetric test: a diagnostic test of certainty in carpal tunnel syndrome]. PMID- 2967482 TI - [Double suborbital neuralgie or "hijab syndrome"]. PMID- 2967483 TI - [Complete atrioventricular dissociation in a patient with systemic scleroderma]. PMID- 2967484 TI - [Kaposi's sarcoma after renal transplantation. A potentially severe opportunistic disease]. PMID- 2967485 TI - [Hyponatremia induced by biguanides. Case report]. PMID- 2967486 TI - [Encephalopathy caused by an ifosfamide-mesna combination]. PMID- 2967487 TI - [Post-traumatic breast cancer: 14 cases. Biological interpretation]. PMID- 2967488 TI - [Cryptococcal meningitis in a patient with rheumatoid arthritis]. PMID- 2967489 TI - [Epidemiology and development of chronic renal insufficiency]. PMID- 2967490 TI - [Varioliform gastritis induced by formol inhalation]. PMID- 2967491 TI - [Clinical and social hygiene characteristics of tuberculosis mortality]. PMID- 2967492 TI - [Role of the environment in the epidemiology of tuberculosis]. PMID- 2967493 TI - Effect of alpha-human atrial natriuretic factor on hemodynamics and hormone levels in aldosterone-producing adenoma (3 case reports). PMID- 2967494 TI - Ultrastructural and electron microscope autoradiographic studies of cytotoxic effect of Y-HpD on cancer cell line. PMID- 2967495 TI - Characterization of cDNA encoding human placental anticoagulant protein (PP4): homology with the lipocortin family. AB - A cDNA library prepared from human placenta was screened for sequences encoding the placental protein 4 (PP4). PP4 is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade. Partial amino acid sequence information from PP4 derived cyanogen bromide fragments was used to design three oligonucleotide probes for screening the library. From 10(6)independent recombinants, 18 clones were identified that hybridized to all three probes. These 18 recombinants contained cDNA inserts encoding a protein of 320 amino acid residues. In addition to the PP4 cDNA we identified 9 other recombinants encoding a protein with considerable similarity (74%) TO PP4, which was termed PP4-X. PP4 and PP4-X belong to the lipocortin family, as judged by their homology to lipocortin I and calpactin I. PMID- 2967496 TI - The cross-pathway control gene of Neurospora crassa, cpc-1, encodes a protein similar to GCN4 of yeast and the DNA-binding domain of the oncogene v-jun-encoded protein. AB - Expression of the gene cpc-1 is required for cross-pathway-mediated regulation of amino acid-biosynthetic genes in Neurospora crassa. We have cloned cpc-1 and present an analysis of its structure and regulation. The cpc-1-encoded transcript contains three open reading frames, two of which are located in the 720 nucleotide leader segment preceding the cpc-1 coding region. The two leader open reading frames, if translated, would produce peptides 20 and 41 residues in length. The deduced amino acid sequence of the cpc-1 polypeptide, CPC1, contains segments similar to the DNA-binding and transcriptional activation domains of GCN4, the major cross-pathway regulatory protein of yeast. The structural and functional similarities of CPC1 and GCN4 proteins suggest that cpc-1 encodes the analogous transcriptional activator of N. crassa. Messenger RNA measurements indicate that cpc-1 is transcriptionally regulated in response to amino acid starvation. The segment of CPC1 similar to the DNA-binding domain of GCN4 also is similar to the DNA-binding domains of the avian sarcoma virus oncogene-encoded v JUN protein and human c-JUN protein. PMID- 2967497 TI - 3-Hydroxyanthranilate oxygenase activity is increased in the brains of Huntington disease victims. AB - An excess of the tryptophan metabolite quinolinic acid in the brain has been hypothetically related to the pathogenesis of Huntington disease. Quinolinate's immediate biosynthetic enzyme, 3-hydroxyanthranilate oxygenase (EC 1.13.11.6), has now been detected in human brain tissue. The activity of 3 hydroxyanthranilate oxygenase is increased in Huntington disease brains as compared to control brains. The increment is particularly pronounced in the striatum, which is known to exhibit the most prominent nerve-cell loss in Huntington disease. Thus, the Huntington disease brain has a disproportionately high capability to produce the endogenous "excitotoxin" quinolinic acid. This finding may be of relevance for clinical, neuropathologic, and biochemical features associated with Huntington disease. PMID- 2967499 TI - Platelet aggregation and vasopressin receptors in patients with diabetes mellitus. AB - Plasma vasopressin, vasopressin-induced platelet aggregation, and platelet vasopressin receptors were investigated in 10 normal subjects and 14 diabetic patients free of microangiopathy. Basal plasma vasopressin concentration was identical in two groups. Platelet aggregation induced by vasopressin as well as by epinephrine was not significantly altered in the diabetic patients. However, exploration of platelet V1-vasopressin receptors revealed in the diabetic group a dramatic reduction in the number of binding sites without alteration of the receptor affinity for tritiated vasopressin. Thus vasopressin-induced platelet aggregation in uncomplicated diabetes mellitus remains normal despite a decrease in the number of vasopressin receptors presumably due to alterations of the platelet membrane structure. PMID- 2967498 TI - Cis-acting sequences that modulate atrial natriuretic factor gene expression. AB - Nucleotide sequences necessary to direct transcription of the gene encoding atrial natriuretic factor (ANF) in neonatal and fetal hearts have been defined by using expression of the prokaryotic marker gene chloramphenicol acetyltransferase (CAT) as a functional assay. Hybrid ANF-CAT genes were introduced into primary cultured cardiocytes by electroporation. A 3.4-kilobase (kb) fragment containing sequences on the 5' side of the ANF gene promoted significant CAT activity in atrial but not ventricular cardiocytes derived from 1-day-old rats. Deletion analysis of putative regulatory regions demonstrated that 2.4 kb of 5' ANF sequences were sufficient for high-level atrial transcription, whereas hybrid genes containing less than 700 base pairs of ANF sequences promoted less CAT activity. Cardiocytes derived from embryonic ventricles expressed the 3.4-kb ANF CAT hybrid gene at levels comparable to atrial cells, suggesting that the nucleotide sequences controlling developmental regulation of ANF expression are contained in this 5' region. Nucleotide sequence analysis of this 3.6-kb region identified segments that may contribute to the regulated expression of the ANF gene. PMID- 2967500 TI - [The antiandrogen cyproterone acetate. Its history from discovery to marketing]. PMID- 2967501 TI - [Effects of dithiocarbamates on the mitotic activity of thymocytes]. AB - The effects of dithiocarbamates on the mitotic activity of thymocytes were studied in vitro and in vivo. The mitotic activity was determined by counting of the metaphases blocked by colchicine. Using thymocytes of mice the mitotic activity in short-time cultures was inhibited (50%) by the following concentrations of the substances under study (EC50-values in mol/l): tetraethylthiuramdisulfide; (1; disulfiram) 4.2.10(-9); diethyldithiocarbamate (2) 3.2.10(-8), Cu-2, 2.6.10(-8), Zn-dimethyldithiocarbamate (3; Ziram) 1.9.10( 8), Zn-ethylene-bis-dithiocarbamate (4; Zineb) 2.3.10(-6). Using rat and guinea pig thymocytes the ED50- values for 1 and 2 were found in the same concentration range as compared with thymocytes of mice. There was no impact on the vitality of cells by the mitosis inhibiting concentrations. In vivo the mitotic activity of thymocytes in mice was decreased only after application of high doses (500 mg/kg i.p.) of 2. After depletion of reduced glutathione (GSH) by the pretreatment of mice with diethylmaleate the effects of 2 were increased in vivo. In vitro GSH (10(-4) mol/l) decreased the mitosis inhibiting activity of 2 in mice thymocytes and the inhibition of the SH-enzyme ecto-ATPase in rat thymocytes. It is supposed that the inhibition of mitosis is due to the reaction of dithiocarbamates with SH groups of microtubules and other functionally important proteines. The formation of tiyl radicals from dithiocarbamates may play a certain role in redox processes which can be influenced by GSH. PMID- 2967502 TI - Stress and anxiety in the disabled patient. AB - This article focuses on patients' feelings of anxiety and fear and the relationship of these feelings to stress. A model is described that identifies four steps in the perception of environmental stress and the emotional responses to it. A second model describes the stages of responses to stress. Both models are applied to the patient experience, and suggestions are offered for basic facilitative physical therapist behaviors. A strong request is made for physical therapists to observe the affective behavior of patients, perceive it in the context of stress, and respond effectively. PMID- 2967503 TI - Powered mobility vehicles as aids in independent locomotion for young children. Suggestion from the field. AB - Both vehicles are good training devices for future mobility. The independent mobility provided by this equipment gave each child a boost of self-esteem and independence. PMID- 2967504 TI - Do we possess adequate models of childhood epilepsies? PMID- 2967505 TI - Sympathetic innervation of blood vessels in rats with aortic constriction hypertension. AB - The adrenergic innervation of blood vessel wall was studied in various vascular beds of adult rats with experimental hypertension induced by the constriction of the aorta between the origins of both renal arteries. A moderate expansion of body fluids was demonstrated in this hypertensive model. The decrease of the density of adrenergic plexus in the vessel wall as well as the diminished catecholamine fluorescence were found only in renal vessels. These changes were pronounced in the left renal artery and vein even if the left kidney was not subjected to elevated blood pressure. Thus the alteration of vascular adrenergic innervation in hypertensive rats is not a consequence of high blood pressure but it seems to be a part of neurohumoral pathogenetic mechanisms. PMID- 2967506 TI - Collagen in the aortae of spontaneously hypertensive (SHR) rats. AB - A biphasic pattern of collagen biosynthesis was found in the aortae of spontaneously hypertensive (SHR) rats; the time course of the rate of biosynthesis is similar to that described in the heart by Sen and Bumpus. In comparison to age-matched controls, collagen biosynthesis is elevated in the SHR rats, diminishes during the first fourteen weeks and rises again at the stage of established hypertension. In the period of established hypertension, the increased rate of collagen biosynthesis was associated with a pronounced rise of the collagen type I to type III ratio. On the other hand, in the pre-hypertensive stage, the proportion of collagen type III clearly exceeds the proportion of collagen type I in SHR rats. PMID- 2967507 TI - Elastic properties of the rat respiratory system related to age. AB - The experiments were performed on male rats of the Wistar strain under urethane anaesthesia (1.3 g/kg i.p.). Changes of oesophageal and tracheal pressures were registered in a group of 30 spontaneously breathing, supine rats, of 295 +/- 13 g average body weight during lung inflations with 1-5 ml of air. In another group of 25 rats of 70 +/- 6 g average body weight (young rats) we made the measurements during inflation with volumes 0.5-2 ml. The measurements were also performed in a group of 10 paralyzed, ventilated rats with 347 +/- 24 g average body weight and inflations 1-5 ml. Compliance of the lungs (CL), chest wall (CW) and of the respiratory system (Crs) was calculated from the linear part of the pressure-volume curve during inflation. The results indicate: 1. Cw is significantly (p less than 0.001) higher in young (134.7 ml.kPa-1.kg-1) than in adult rats (44.1 ml.kPa-1.kg-1). CL (related to body weight) is not significantly different in young and adult rats. 2. Cw is significantly (p less than 0.001) higher than CL. 3. No difference was observed in CLs Cw and Crs between paralyzed and spontaneously breathing animals. PMID- 2967508 TI - Circaseptan rhythm of corticosterone and lipids in the serum of male rats. AB - After adaptation to standard conditions and to a 12:12 h light: dark regimen, groups of young SPF male Wistar rats were killed at 8 h on 17 consecutive days and their serum corticosterone and lipid concentrations were determined. Using a combination of a Fourier harmonic analysis and an analysis of variance, the circaseptan rhythm of corticosterone, total cholesterol and triacylglycerol oscillations was determined; it is unlikely that the nonesterified fatty acid concentration has a 7-day rhythm. The circaseptan variation of metabolic and hormonal indicators is evidently adaptive in character and its existence ought to be taken particularly into account in the evaluation of long-term experiments. PMID- 2967509 TI - Effect of repeated stress and administration of phenobarbital on the lymphoid tissue and on protein metabolism in the lymphocytes of infant and adult rats. AB - Further study of the response to chronic stress stimulation in the early postnatal phase showed that the i.p. injection of physiological saline (stress stimulation) induced lymphopenia, a 50% decrease in the incorporation of 3H leucine into isolated lymphocytes and a decrease in the weight of the thymus in 7 day-old male rats. No such changes were observed in adult animals. If repeated doses of phenobarbital were administered to stressed young rats, however, lymphopenia did not occur and the rate of the incorporation of 3H-leucine into isolated lymphocytes was not different from the control value; the protein content of the lymphocytes was significantly raised, however. In adult animals, phenobarbital increased the rate of incorporation of 3H-leucine into the lymphocytes. The repeated administration of phenobarbital reduced the weight of the thymus in both young and adult animals, but a decrease in spleen weight was recorded only in the young animals. A single i.p. injection of ACTH or dexamethasone caused lymphopenia and slowed down the incorporation of 3H-leucine into the lymphocytes of both young and adult animals. The results show that the striking decrease observed in the rate of the liver metabolism of corticosterone in suckling young rats not injured by repeated stress stimulation is accompanied by significant changes in the lymphoid tissue. PMID- 2967510 TI - Effect of the antimuscarinic agent pirenzepine on the in vivo biliary secretion of dogs in response to various stimuli. AB - Pirenzepine is known to be an antiulcer drug with antimuscarinic activity. The present work shows the effect of pirenzepine dihydrochloride on biliary secretion in dogs under normal conditions and after the application of different nervous and humoral stimuli. Pirenzepine (3 mg/kg) was orally administered to unanaesthetized dogs one hour before feeding. This treatment diminished the increase in biliary secretion as well as the intracholedochal pressure that usually followed feeding. On the other hand, a 0.75 mg/kg dose of the antimuscarinic drug intravenously administered to anaesthetized dogs, significantly reduced the increase in intracholedochal pressure produced after the injection of acetylcholine or cholecystokinin (CCK-PZ). Finally, the same dose of pirenzepine eliminated the effect of vagal electrical stimulation on intracholedochal pressure. These results suggest that the effect of pirenzepine on biliary secretion is mainly due to its action on the emptying of the gallbladder. PMID- 2967512 TI - The influence of mixed venous pO2 on the natriuretic activity of cat plasma. AB - Plasma was taken from anaesthetised cats with high or low mixed venous pO2. Different plasma fractions were then administered to anaesthetised rats to examine the effect on sodium excretion. It was found that only the low molecular weight fraction of the plasma from cast with high mixed venous pO2 showed any natriuretic activity. A lipid soluble extract from the same plasma also increased sodium excretion. It is concluded that an increase in mixed venous pO2 in anaesthetised cats leads to the release or activation of a natriuretic factor, which has a molecular weight below 700 and is lipid soluble. PMID- 2967513 TI - Differential DC amplifier for recording small and fast concentration changes with ion-selective microelectrodes. PMID- 2967511 TI - Effects of copper, cadmium and chromium cations on the freshwater fish Clarias batrachus L. AB - The data on the effects of cations such as Cu2+, Cd2+ and Cr6+ on the changes in the biochemical parameters in a freshwater fish, Clarias batrachus L., showed an increase of the protein content in the liver, kidney, stomach, intestine, testis and ovary, and a decrease in the muscle after Cu2+ and Cd2+ treatment as compared with control data; but the Cr6+ did not cause any changes of protein concentration in the kidney and testis. The administration of Cu2+ and Cd2+ increased the concentration of free amino acids in all the fish organs, whereas the Cr6+ did not changes this concentration in the muscle. A decrease in dry weight, and an increase in tissue permeability after these treatments were recorded in all the organs studied. In general, the above biochemical parameters of the organs were affected by treatments of the above cations in the following order: Cd greater than Cu greater than Cr over control values of C. batrachus, and their effects were markedly pronounced in the liver and kidney, followed by the intestine, stomach, muscle, testis and ovary in this species. PMID- 2967514 TI - On the role of calcium in the mechanism of ADH action. AB - With an increased influx of Ca2+ in the cytoplasm, the response of cells to ADH in the urinary bladder of the frog was lowered by addition of ionophore A23187 from the side of the basolateral cell membrane, but inhibited when it was added from the apical cell membrane. The removal of calcium by EGTA from the serosal surface was accompanied by a sharp increase of osmotic permeability not only to water, but also to inulin; while when calcium was removed from the mucosal surface of the urinary bladder, osmotic permeability was not changed. After being added to the Ringer solution from the outer surface of the apical cell membrane, the inhibitors of Ca2+ channels (verapamil, Ni2+, Mn2+, Co2+) decreased the effect of ADH. These data indicate that Ca2+ applied onto the outer surface of apical plasma membrane plays an important role in the action of ADH. PMID- 2967515 TI - Social perceptions of the effects of Down syndrome facial surgery: a school-based study of ratings by normal adolescents. AB - This study examines how 277 normal adolescents from five Israeli schools perceive the Down syndrome face before and after plastic surgery. A seven-point Likert scale was used to rate slides of normal and Down syndrome faces on four dimensions. Down syndrome patients were seen as less attractive, intelligent, good-hearted, and socially appealing than normal individuals. The slides were in nonapparent order and contained preoperative and 1-year postoperative views of eight plastic surgical patients. Paired t tests were used to examine operative changes, and all four dimensions showed overall postoperative improvement (p less than 0.001), but case-to-case variation was considerable. A linear relationship was found between change in appearance ratings and change in intelligence ratings. Findings suggest that when improvements in facial appearance are realized, peer normal social perceptions of the Down syndrome child may be enhanced. The relationship between school placement, intellectual level, and surgical decision making is discussed. PMID- 2967516 TI - The extended latissimus dorsi flap in repair of anterior abdominal wall defects. AB - Large abdominal wall defects (ventral hernias) can be difficult to repair. Some defects are not amenable to primary repair or the use of synthetic mesh because of repeated recurrence or wound infection. In complicated situations such as that mentioned above, the extended latissimus dorsi muscle flap has been used to repair upper and middle abdominal wall defects. This method has been utilized in six patients, and there has been no recurrence of the defect or evidence of a lumbar hernia. The follow-up has been from 7 to 66 months. The extended latissimus dorsi muscle flap has proven to be an excellent alternative in the repair of complicated abdominal wall defects. PMID- 2967517 TI - Hormonal basis during pregnancy for the onset of maternal behavior in the rat. AB - This article reviews the current state of our knowledge about the hormonal basis of maternal behavior in the rat. Considered are the ovarian hormones estrogen and progesterone, the pituitary hormones beta-endorphin and prolactin, and the hormone oxytocin, secreted by several hypothalamic nuclei and associated brain regions. The hormones of pregnancy, estrogen and progesterone, prime the female to respond to a terminal rise in estrogen that stimulates a high level of maternal responsiveness even before parturition begins. Studies on the role of prolactin, using hypophysectomy, prolactin release blockers and anterior pituitary and prolactin replacement, indicate that prolactin is required for the ovarian hormones to be effective in stimulating maternal behavior. During the latter half of pregnancy, placental lactogen may displace prolactin in this role. Although prolactin serves as a chronic stimulus for maternal behavior, it also may act over a short period. Oxytocin stimulates maternal behavior in a specific strain of rat, but not in other strains, and only when administered introcerebroventricularly (ICV) in estrogen-primed females. The decline in the high brain levels of beta-endorphin around parturition has been proposed as a requirement for the onset of maternal behavior; morphine blocks the onset of maternal behavior and disrupts ongoing maternal behavior and maternal aggression in lactating females. However, blocking beta-endorphin action at parturition interferes with pup cleaning and eating of the placenta as well. PMID- 2967518 TI - Fawn hooded rats are subsensitive to the food intake suppressant effects of 5-HT agonists. AB - The food intake suppressant effects of three serotonin agonists, m-CPP (a selective 5-HT1B agonist), 8-OHDPAT (a selective 5-HT1A agonist) and fenfluramine (a 5-HT releasing agent) were compared in three different rat strains: Wistar, Sprague-Dawley (SD) and Fawn-Hooded (FH) rats. Administration of all three serotonin agonists produced dose-dependent decreases in 1 h food intake in all three strains. FH animals were significantly less sensitive to the food intake suppressant effects of all three serotonin agonists than either Wistar or SD rats. Body weight gain over the 9-week course of the study was also significantly less in FH animals than either Wistar or SD animals. These findings support some other data that Fawn Hooded rats, a strain with a peripheral platelet serotonin storage disorder, also possess altered central nervous system serotonergic function. PMID- 2967519 TI - [Response to vaccination against the etiologic agent of viral hepatitis B (HBV) in a population at risk]. AB - We report the results of the vaccinations against hepatitis B carried out with vaccine H-B-VAX of the commercial firm (Merck Sharp and Dohme), on 102 U.S.L. 32 hospital workers in Portomaggiore (Ferrara, Italy). The vaccine rendered 92% of the subjects inoculated immune and the incidence of secondary effects was low. PMID- 2967520 TI - Tissue repair and repopulation in the tumor bed effect. AB - These experiments were designed to study the kinetics and magnitude of cell repair and repopulation in tissues whose damage results in the tumor bed effect. The right hind thighs of mice were irradiated with single doses or two equal gamma-ray fractions. Interfraction intervals ranging from 30 min to 24 h (to measure the kinetics of repair from sublethal damage) and 6 and 12 weeks (to determine the extent of repopulation) were used. One day after the second radiation dose 5 X 10(5) FSA tumor cells were inoculated into the center of the irradiated field. Radiation dose-response curves were obtained by calculating the time required for tumors to reach 12 mm diameter. No recovery occurred within 6 h of the radiation delivery as measured by this assay. Some recovery, 3.2-4.6 Gy above a single radiation dose, occurred when the interval between two fractions was 24 h. With increasing interfraction intervals of 6 and 12 weeks further dose sparing occurred in the amount of 5.0-6.9 and 7.5-8.3 Gy, respectively. The data suggest that repopulation is the major contributor to the radiation dose-sparing recovery of stromal tissue and that some proliferative response may occur as early as 1 day after the first irradiation. PMID- 2967521 TI - Prophylaxis of hepatitis B infection. PMID- 2967522 TI - Tumor immunotherapy by local injection of interleukin 2 and non-reactive lymphocytes. Experimental and clinical results. PMID- 2967523 TI - Cellular mechanisms of tumor rejection in vivo and enhanced induction of anti tumor protective immunity applicable to tumor-specific immunotherapy. PMID- 2967524 TI - Down's syndrome and the molecular biology of chromosome 21. PMID- 2967525 TI - [The Nazi physicians Aquillin Ullrich and Heinrich Bunke in the legal process concerning euthanasia]. PMID- 2967526 TI - [Dermatitis seborrheica and HIV infection]. PMID- 2967527 TI - [Continuous-wave ultrasonography in the diagnosis of portal hypertension]. PMID- 2967529 TI - [Aging change of carotid blood flow]. PMID- 2967528 TI - [Characterization and physiological development of receptors for insulin-like growth factors I and II (IGFs) in the mammary gland of ewes]. AB - Membrane preparations were obtained by differential centrifugations of ewe mammary gland homogenates. These membrane preparations contained specific receptors for IGF1 and IGF2 which possess high affinities for their specific ligands (Ka .5 to 1.5 10(9) M-1). Maximum binding of 125I IGF1 was obtained after 48 h at 4 degrees C. This binding was inhibited by unlabelled IGF1 (ED50 = 14 ng/ml), partially inhibited by high concentrations of insulin (50 micrograms/ml). Prolactin (oPRL), growth hormone (bGH) or relaxin (Rel) were without effect. Maximum binding of 125I IGF2 was obtained after 6 h at 20 degrees C. This binding was inhibited by unlabelled IGF2 (ED50 = 44 ng/ml), partially inhibited by IGF1 (ED50 = 200 ng/ml) and unmodified by INS, PRL, bGH or Rel. Receptor numbers for IGF1 were significantly higher (p less than 0.01) on day 100 of pregnancy (N = 480 +/- 17 fmoles/mg proteins) compared to day 20 of lactation (N = 174 +/- 21 fmoles/mg). The numbers of IGF2 receptors were always higher than those of IGF1 receptors. During pregnancy the numbers of IGF2 receptors (N = 1,860 +/- 157 fmoles/mg) were also higher than during lactation. These results suggest that the mammary gland may constitute a target organ for IGFs. These factors could be involved in the regulation of mammary gland development and during cell differentiation. PMID- 2967530 TI - [Behcet's disease complicated by acute myocardial infarction treated with percutaneous transluminal coronary angioplasty]. PMID- 2967531 TI - [Physio-pathogenesis of the acquired immunodeficiency syndrome]. PMID- 2967532 TI - [Dermatologic manifestations of the acquired immunodeficiency syndrome]. PMID- 2967534 TI - [Renovascular hypertension due to transplantation-induced stenosis of the iliac artery. Successful therapy with transluminal angioplasty]. PMID- 2967536 TI - [Dwelling tomorrow]. PMID- 2967535 TI - [Crown-root restorations with plastic materials. Significance of mixed glass metal cements]. PMID- 2967533 TI - Sports medicine and the wheelchair athlete. AB - International competitions for the wheelchair-confined are now a major feature of the world of sport. They are helpful in improving both mood state and physiological function, while improving long term prognosis. Immediate medical problems are much as in other types of competition, but there are also specific problems (bladder infections, pressure sores, intolerance of environmental extremes, and injuries related to wheelchair use). Disability classification, based on the anatomical or functional level of a lesion, provides a reasonably fair basis for competition. Most of the functional data to date relate to strength (isometric and isokinetic) and aerobic power (measured in a wheelchair or on an arm ergometer). While the inactive patient is often severely limited, wheelchair athletes may have a greater functional capacity than sedentary normals. The principles of training for the wheelchair-confined are much as in the able-bodied, although because the arm muscles are small, much of the training response may be peripheral rather than central. The margin between an effective stimulus and overtraining is also reduced. Involvement in a training programme not only increases physiological function, but also counters depression, increasing the subject's sense of self-efficacy. Biomechanicians are now contributing increasingly to wheelchair sport, improving the design of competitive wheelchairs, improving the mechanical efficiency of participants, and helping to reduce the risks of injury. The benefits of wheelchair sport are now clearly established, and family physicians should do more to encourage the involvement of the wheelchair-confined. PMID- 2967537 TI - Interleukin-2-induced proliferation of CD4-CD8- human thymocytes. In vitro expression of CD3 and CD8 antigens and cytolytic activity. AB - Human thymocytes lacking both CD4 and CD8 differentiation antigens were prepared by treating total thymocyte suspensions with a mixture of anti-CD4 and anti-CD8 monoclonal antibodies and complement. The resulting populations contained less than 2% CD4+, CD8+ or WT31+ cells and variable percentages (less than 20%) of CD3+ cells. These cell populations were cultured in recombinant IL-2 in the presence of peripheral blood mononuclear cells as feeder cells. Cells underwent extensive proliferation accompanied by a progressive increase of CD3+ and CD8+ cells. On the other hand, appearance of neither WT31+, alpha/beta-positive T cell receptor (TCR), nor CD4+ cells could be observed in several independent experiments. Functional analyses revealed the appearance and the progressive increase of cytolytic activity against the natural killer (NK)-sensitive K562 cells as well as the NK-resistant fresh melanoma cells. Experiments of T cell cloning indicated that both the expression of CD8 and CD3 antigens and the appearance of cytolytic activity were consequent to cell maturation occurring at the level of CD4-CD8- non-cytolytic cell precursors. In these experiments, more than 30% of cells underwent clonal expansion and all the clonal progenies obtained displayed cytolytic activity and expressed the CD3+WT31- surface phenotype. The expression of CD8 was variable, whereas no CD4+ clones could be obtained. Cells expressing such surface phenotype are known to belong to the TCR gamma-positive T lymphocyte subset lacking the typical alpha/beta TCR and thus appear to be the only T cell type capable of in vitro proliferation and maturation under easily reproducible culture conditions. PMID- 2967538 TI - Look at people--not handicaps. PMID- 2967539 TI - T and B cell specific immune responses to purified protein derivative in the cerebrospinal cavity may be maintained and regulated independently of systemic immune control. AB - In vivo activated T cells could be isolated from cerebrospinal fluid (CSF) of a patient suffering from chronic meningitis of unclear origin. Although the patient's skin reactivity to purified protein derivative (PPD) was negative, and peripheral T cells did not proliferate to this antigen in vitro, the majority of T cell clones from CSF specifically recognized PPD on either autologous or allogeneic HLA class II compatible macrophages. Remarkably, peripheral blood mononuclear cells potently suppressed the PPD-specific proliferative responses of healthy donors. The selective enrichment of oligoclonal IgG in the CSF but not in the patient's serum further indicated T and B cell responses lacking systemic feedback control. Analyses of a persisting immune stimulation in the CSF provide a potent diagnostic tool and may explain neurological complications as observed in a number of autoimmune diseases and chronic infections. PMID- 2967540 TI - Role of accessory cells in the activation of pure T cells via the T cell receptor CD3 complex or with phytohaemagglutinin. AB - The ability of different subpopulations of blood mononuclear cells to serve as accessory cells in the activation of CD4+ and CD8+ T cells via Ti-CD3 or with phytohaemagglutinin (PHA) was studied. Pure CD4+ or CD8+ T cells did not respond to particle-bound anti-CD3 monoclonal antibodies (MoAb) or PHA, whereas responses were seen when non-T cells served as accessory cells. Removal of class II positive cells from peripheral blood mononuclear cells (PBMC) or from non-T cells diminished, but did not completely abolish, the responses in both T cell subsets, indicating that the accessory cells are mainly found among the class II-positive cells. However, the class II molecules themselves were not involved, as demonstrated in antibody-blocking experiments. Removal of monocytes decreased the ability of non-T cells to serve as accessory cells for both CD4+ and CD8+ cells in PHA activation. In contrast, the removal of monocytes resulted in an enhanced activation by anti-CD3 MoAb in CD4+ T cells, while the activation of CD8+ T cells was less affected. Positively selected B cells were effective accessory cells in anti-CD3 and PHA activation. Furthermore, Epstein-Barr virus (EBV)-transformed B cell lines were very potent accessory cells both in anti-CD3 and PHA activation of T cells, and showed the strongest accessory cell function observed in this system on a per cell basis. PMID- 2967541 TI - Regulation of prostatic growth. Growth-limiting mechanisms in the rat ventral prostate. AB - The possibility that a prostatic chalone is a factor regulating growth of the gland was investigated in rats submitted to hemiprostatectomy to reduce the level of a hypothetical circulating chalone. No evidence of regeneration was found 4 weeks postoperatively. Orchiectomized rats with intrasplenic, intramuscular or intraprostatic implants of ventral prostatic tissue from other orchiectomized rats, were given androgen. The cell divisions in the prostatic tissue did not end until the total cell number was 150-170% of that in control rats. These results indicate that other factors than a prostatic chalone have a dominant role in the mechanism limiting prostatic growth. As introduction of an artificial stroma led to increased prostatic growth, it appears likely that the fibromuscular stroma is one of the factors controlling growth of the prostatic epithelial cells. PMID- 2967542 TI - [Regression of myocardial hypertrophy in aortic valve disease following aortic valve replacement]. AB - In patients with aortic valve disease successful valve replacement leads to a significant decrease in left ventricular angiographic mass, which 14-15 months postoperatively ranges between 31 and 37% of the preoperative value. Postoperative normalization of angiographic mass occurs in 2/3 of the patients with aortic stenosis and in 1/3 of the patients with aortic insufficiency. The degree of preoperative hypertrophy determines whether angiographic mass does or does not normalize after valve replacement. In patients with aortic stenosis and combined lesions, left ventricular end-diastolic wall thickness decreases postoperatively and remains unchanged in the patients with aortic insufficiency. Muscle fiber diameter decreases significantly in the first two years after valve replacement and thereafter remains unchanged. The upper limit of normal (20 mu) is not reached. Relative interstitial fibrosis increases early after surgery whereas left ventricular fibrous content remains unchanged during the first two years after valve replacement. Five years postoperatively there was a significant decrease in left ventricular fibrous content both in patients with aortic stenosis and with aortic insufficiency. Hence there appears to be no longer any validity in the earlier concept that a once established interstitial fibrosis in patients with concentric and eccentric hypertrophy is irreversible after removal of the abnormal hemodynamic burden. PMID- 2967543 TI - [Duplex sonography in the diagnosis of peripheral arterial circulation disorders]. AB - From January 1984 to June 1986, 621 duplex scans of the arteries of the lower extremities were performed. 54.8% of the examinations were performed for clinical reasons. In 45.2% the indication was strictly scientific. Clinical and non invasive examinations including pulse-volume recordings and Doppler pressure measurements preceded duplex-scanning. The analysis shows that duplex scanning may replace arteriography in patients with localized stenosis and short occlusions where catheter therapy is possible. Patients with insignificant stenosis or long occlusion may be selected for conservative treatment without arteriography. PMID- 2967544 TI - Examination of the fetal heart in the fetus with intrauterine growth retardation using M-mode echocardiography. PMID- 2967545 TI - Mentally retarded children of eastern India: a biosocial study. PMID- 2967546 TI - Hypertensive concentric left ventricular hypertrophy: when is ventricular ectopic activity increased? AB - The Framingham Study has indicated that patients with left ventricular hypertrophy (LVH) have a greater risk of cardiovascular complications and sudden death than subjects with a normal heart. We have previously demonstrated that ventricular ectopy was more prevalent and complex in hypertensive patients with LVH by electrocardiographic (ECG) criteria than in those without ECG evidence of LVH. The present study was designed to detect and quantify ventricular dysrhythmias in hypertensive patients with early concentric LVH by echocardiography but without LVH by ECG criteria. Continuous ambulatory ECG tracings were recorded for 24 hours in 94 patients with essential hypertension: 37 without LVH, 26 with concentric LVH by echocardiographic but not ECG criteria, and 31 with LVH on both echocardiography and ECG. Patients with LVH by ECG criteria had significantly more premature ventricular contractions (P less than .001) and more complex (higher Lown's class) ventricular ectopy (P less than .001) than hypertensives without LVH or with LVH only by echocardiographic criteria. Prevalence and complexity of ventricular ectopic activity, however, was not affected by mild to moderate concentric cardiac hypertrophy detected echocardiographically. We conclude that unlike LVH shown by ECG, early hypertensive concentric LVH detected echocardiographically is not associated with increased electrical irritability of the myocardium. PMID- 2967547 TI - [Characteristics of the working and living conditions of women district therapists]. PMID- 2967548 TI - [Joint activity of medico-sanitary units and the administration and social organizations of large industrial plants in protecting the health of workers]. PMID- 2967549 TI - The changing face of coronary angioplasty. PMID- 2967550 TI - Ethical considerations in fitness and risk evaluations. AB - Physicians and other health care workers involved in the delivery of occupational health care can be confronted with difficult questions about fairness, justice and professional ethics. These issues and other ethical considerations are discussed. PMID- 2967551 TI - Low back pain: risk evaluation and preplacement screening. AB - As screening tests in a currently asymptomatic population, all of the available methods for predicting low back pain and disability have serious technical, ethical and legal limitations. From a technical point of view, none of the predictive tests appear to have sufficient sensitivity or specificity to justify routine usage. The most sensitive indicator (a past history of low back pain) lacks reliability and specificity. Muscle strength-testing may be predictive of future musculoskeletal injury as part of a well-designed program that considers specific job demands in relation to specific worker capabilities. Unfortunately, the conditions necessary for a well designed program--a large number of predictable high-risk jobs that have measurable specific demands that can be reproduced reliably in a testing situation--are rarely met. In the future, the use of computer-assisted multivariate models analogous to those used in the prediction of cardiovascular risk may be capable of integrating information about an individual's medical history, physical exam, physical capacity and other tests with specific job requirements to give us a more accurate prediction of the future risk of back pain and disability. If such predictive models are ever developed and verified, it would then be appropriate to examine various interventions and their effectiveness in modifying risks for the population and the individual worker. From a legal point of view, all of the techniques described hold potential for significant discrimination against legally protected groups. Making employment decisions with regard to a past history of low back pain or on the basis of an x-ray will lead to systematic age discrimination and discrimination against the handicapped. The use of muscle strength-testing will systematically discriminate against women and certain ethnic groups. The ethical implications of predictive screening for low back pain clearly depends on what is done with the information that is garnered from such tests. If the information is used only to make a safe job placement for an individual, and this placement does not affect the individual's salary or future job possibilities, then the testing program may have a net social value to the extent that it leads to true prevention of low back pain and disability. If the tests are used merely to reduce employer liability by refusing employment to those who are thought to be at "high risk," the technical, legal and ethical limitations will far outweigh any perceived benefits. PMID- 2967552 TI - Dermatologic considerations in worker fitness evaluation. AB - Dermatoses account for a very high percentage of all occupational diseases. The authors review individual factors that increase the risk of work-related skin disease and present information to assist in the evaluation of individuals in preemployment situations and for guidance and counseling prior to occupational choices. PMID- 2967553 TI - [Anti-hepatitis B vaccination in a hemodialysis service]. PMID- 2967554 TI - Transcutaneous incorporation of nonabsorbable monofilament sutures. AB - In rats and guinea pigs, transcutaneous through and through mattress sutures of different materials (silk, nylon and Prolene [polypropylene]) were tied under tension to plicate the abdominal wall or the scrotal wall. Knots were either buried subcutaneously or left outside. Monofilament sutures, with buried knots, cut through and became completely incorporated as the plicated normal tissue flattened out to a normal appearance in about three weeks. Silk sutures and bulky outside knots left suppurating sinuses. Histologic examination at seven weeks confirmed the incorporated nylon or Prolene mattress sutures were surrounded by a sheath of clean scar tissue trailing as a curtain back to the cut through skin. It is concluded that sutures of monofilament nylon or Prolene, with buried knots, can be tied under tension to include outside skin and will cut through cleanly to become incorporated without sinus or suppuration. Such incorporated sutures and their trailing curtains of clean scar tissue can provide an internal buttress against soft tissue displacement with potential clinical application. PMID- 2967555 TI - Effect of pharmacologic agents on the function of the hypothermically preserved dog kidney during normothermic reperfusion. AB - We examined how a combination of pharmacologic agents ("rescue" agents) affect the function of hypothermically preserved dog kidneys at the time of reperfusion. Dog kidneys were preserved either by simple cold storage in EuroCollins' solution for 24 or 48 hours or by continuous perfusion at 5 degrees C in Belzer's gluconate-hydroxyethyl starch solution for as long as 5 days. After preservation, renal functions were measured with the isolated perfused kidney model. Kidneys were reperfused at normothermia either with or without the addition of a combination of rescue agents to the reperfusion medium. The rescue agents studied were allopurinol (1 mmol/L); superoxide dismutase (32,000 U/L); catalase (137,500 U/L); dimethylthiourea (3 mmol/L); glutathione (3 mmol/L); desferrioxamine (0.2 gm/L), for protection against O2 free radical injury and lipid peroxidation injury; verapamil (25 mg/L), as a Ca channel blocker; and ATP-MgCl2 (0.3 mmol/L), to stimulate energy metabolism. The renal functions we measured were glomerular filtration rate (GFR) (creatinine clearance), urine production, perfusate flow, urinary protein concentration, Na reabsorptive capacity, and tissue concentrations of ATP, K, and total tissue water. GFR was reduced by 75% to 90% after all periods of preservation, and the rescue agents had no effect on GFR. Sodium reabsorption was reduced from 98% to a range of 40% to 50% after 48 hours of cold storage or 5 days of machine perfusion and was not increased by rescue agents. There was a time-dependent increase in the amount of urine protein that was not affected by rescue agents. The addition of rescue agents did not affect total tissue water or concentrations of ATP or K in kidneys after normothermic reperfusion. These results demonstrate that pharmacologic agents previously suggested to suppress reperfusion damage in kidneys are not effective in this model. Therefore it is likely that kidneys damage occurs primarily during preservation, which suggest that optimal function on reperfusion calls for the development of better methods of preservation. PMID- 2967556 TI - Levels and plasma distribution of free and C4b-BP-bound protein S in human fetuses and full-term newborns. AB - Levels and plasma distribution of protein S were measured on umbilical cord plasmas from 25 normal full-term newborns and 25 normal fetuses which were between 20 and 31 weeks of gestation. Fetal blood samples were obtained by direct puncture of the umbilical vein under high resolution real-time ultrasound. Total and Free protein S levels were found to be lower in all fetuses and newborns as compared to normal adults. The calculated ratio Total protein S/Free protein S and crossed immunoelectrophoresis patterns indicate that protein S circulates essentially in the free form in fetuses and newborns. These data may be explained by the low levels of C4b-binding protein observed at these stages of development. PMID- 2967557 TI - The effects of oestrogen administration on the plasma free protein S and C4b binding protein. AB - This study was undertaken to evaluate the effects of oestrogen administration (low dose as an oral contraceptive or higher dose as a hormone replacement therapy) on the levels of plasma free protein S and C4b-binding protein. The participants were 59 women aged 18-49 years, divided into 2 groups: A and B. Group A was composed of 22 post-menopausal women on a hormonal replacement therapy programme (HRT) consisting of 2 mgs daily oestradiol valerate for 21 days. Group B was divided into subgroup B1: 18 women who had been on oral contraceptive for at least one year and subgroup B2 (control): 17 women who were not pregnant and not taking any oral contraceptive. In this study were also included two young women who both suffered from severe thromboembolic disease a few months after initiation of oral contraceptive. The first was 25 years old, with congenital moderately decreased prekallikrein (activity and antigen 40% and 45% respectively) and the second was a 21 year-old woman with congenital moderately decreased plasminogen activity and antigen 45%). In both cases, family members with similarly reduced levels of prekallikrein (PK) and plasminogen (PLG) respectively were free from any thromboembolic disease and had normal protein S levels. In Group A, 22 women at the end of the first cycle of treatment, had lower levels of free protein S (p less than 0.001) than before the initiation of HRT. In subgroup B1, the levels of free protein S were found to be significantly lower than in subgroup B2 (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967558 TI - Therapeutic in vivo and in vitro effects of argon dye laser and hematoporphyrin derivative in lung cancer. AB - In clinical and experimental study the therapeutic efficacy of argon dye laser irradiation with hematoporphyrin derivative (HpD) was evaluated in lung cancer. A total of 14 lung cancer cases including 12 squamous cell, 1 adeno- and 1 small cell carcinomas were irradiated superficially 48 hours or more after i.v. injection of 3 mg/kg of HpD (100-300 mW, 20-30 min.). Human adenocarcinoma cells implanted subcutaneously into nude mice were photoirradiated (200 mW, 20 min.) 48 h after i.p. injection of HpD. The in vitro effect of phototherapy was studied in the same cell line after incubation in medium containing HpD compared to untreated, only irradiated or only in HpD incubated cells. Among 3 early stage squamous cell carcinoma cases 2 complete and 1 partial remissions were obtained. Among 11 cases including 10 with advanced and 1 with recurrent disease 7 demonstrated partial remission. In vivo, two of 9 mice had a complete tumor remission. In the in vitro study, tumor cells incubated in 30 micrograms HpD/ml showed severe cytotoxic effects resulting in cell death 12 hours after photo irradiation, whereas cells incubated in 30 micrograms HpD/ml only regenerated after initial cytotoxic reaction. Laser irradiation only had no effects. HpD phototherapy demonstrated a considerable antitumor efficacy and must be considered as one of the promising endoscopic treatments in cases with early stage primary lung cancer. PMID- 2967559 TI - Surgical treatment of coronary aneurysm developed after PTCA. AB - Two patients who developed coronary aneurysm at the site of Percutaneous Transluminal Coronary Angioplasty (PTCA) in proximal LAD accompanied by severe re stenosis just proximal to the aneurysm are described. Both patients underwent Coronary Arterial Bypass Grafting (CABG) to distal LAD to stop anginal attacks refractory to any anti-anginal drugs and to prevent a rupture of a coronary aneurysm. After the operation the anginal attacks disappeared and no trace of coronary aneurysm was visible in the coronary angiogram. PMID- 2967560 TI - [Concern over comprehensive care. Caregiver of invalid older people]. PMID- 2967561 TI - [Reactive changes in the heat-resistant muscle tissue of lake frogs during heat acclimation]. AB - A study was made of the effect of heat acclimation of animals on the individual changes in the muscle heat resistance in populations of Rana ridibunda belonging to European or Asiatic groups of the species. The relationship between the levels of heat resistance of muscle cells and the value of its changes are the same in the populations of these groups. It is believed that this relationship forms on the level higher than populational one. PMID- 2967562 TI - Acne vulgaris in adolescence. PMID- 2967563 TI - [Percutaneous transluminal coronary angioplasty in patients with angina pectoris]. PMID- 2967564 TI - [Exanthema in 2 patients with ulcerative colitis treated with Pentasa]. PMID- 2967565 TI - [Occupational dermatology studies in the occupational medicine clinic in Arhus. A report of the pattern of referral and results]. PMID- 2967566 TI - [Occupational dermatology in Denmark. Comment on the Arhus model]. PMID- 2967567 TI - [Epidural steroids in spinal pain with nerve root irritation]. PMID- 2967568 TI - [Lumbar traction in acute lumbago/sciatica]. PMID- 2967569 TI - [Arrhythmogenic right ventricular dysplasia]. PMID- 2967570 TI - [Development of new drugs based on the structure of natural polypeptides]. PMID- 2967571 TI - [The heart, an endocrine gland. Therapeutic implications]. PMID- 2967572 TI - [Expression of the gene for the atrial natriuretic factor: a marker for ventricular hypertrophy]. PMID- 2967573 TI - [Atrial natriuretic factor in experimental hypertension in the rat]. PMID- 2967574 TI - [Orthostatic hypotension: new therapeutic approaches]. PMID- 2967575 TI - Phosphofructokinase activity in normal diploid mice during development and in trisomy 16 fetal mice. AB - Although several of the genes mapped to human chromosome 21 have been assigned to mouse chromosome 16, it has not yet been possible to do this for the gene for the phosphofructokinase liver type subunit (PFKL). The goal of this study was to determine if there is a 1.5-fold increase of the PFK activity in fetal trisomy 16 mice, which, if present, would be indicative of a gene dosage effect. However, rather than an increase, an almost 100% decrease of the mean PFK activity was observed in fetal trisomy 16 liver at both 14 and 17 days of gestation when compared to littermate controls. This is the first biochemical abnormality detected in trisomy 16 fetal liver. In contrast, no significant differences in the mean PFK activity in homogenized whole fetus or brain trisomy 16 and diploid controls were observed. A developmental maturational effect of the PFK activity was observed in fetal liver from normal diploid mice, with a 3-fold increase of the activity from day 14 to day 18 of gestation and a further 2-fold increase to adulthood. The decreased PFK activity in fetal trisomy 16 liver may therefore be the result of delayed maturation of the liver in trisomic fetuses. PMID- 2967576 TI - Lack of correlation between the grade of methacholine-induced bronchial hyperreactivity and ipratropium bronchodilation in asthmatics. AB - In 46 never-smoking randomly chosen patients with non-allergic asthma, 40 to 60 years old, a methacholine hyperreactivity test and lung function tests were performed after inhalation of different doses of ipratropium bromide (IB). The grade of hyperreactivity was measured as the cumulative dose of methacholine necessary to produce a decrease in the forced expiratory volume in one second of 20% of the lowest post-NaCl value (PD20). The following lung function tests were carried out: Lung volumes, ventilatory capacity including flow-volume curves, airway resistance and nitrogen single-breath wash-out test. The bronchodilator effect, measured as a change in the different lung function tests for different doses of IB given (0.08 mg, 0.15 mg and 0.25 mg), was correlated to the grade of hyperreactivity (PD20 dose). No or only a slight correlation was found between the grade of methacholine-induced hyperreactivity and the bronchodilator effects of the different doses of IB. These results indicate a lack of correlation between an anticholinergic bronchodilator effect and the grade of methacholine induced bronchial hyperreactivity, or possibly an insensitivity of the above mentioned methacholine test. PMID- 2967577 TI - [Patient survival over 20 to 25 years after mitral commissurotomy]. AB - The authors report that 282 of 457 patients (62%) are alive for 20-25 years after mitral commissurotomy. The twenty-five-year survival is higher in women (70%) than in men (30%). PMID- 2967578 TI - [Improvement in the quality of student training in surgery]. PMID- 2967579 TI - A rapid micro-method for the study of antibody-mediated killing of bacteria, with specific application to infection of sheep with Pasteurella haemolytica. AB - A micro-titration plate bactericidal assay was developed to measure complement dependent antibody-mediated killing of Pasteurella haemolytica. Sera and lung washings from specific pathogen-free (SPF) lambs convalescent from a challenge with live, virulent P. haemolytica were bactericidal in the presence of complement. Similar samples from naive SPF lambs had no such activity. Purified IgG derived from a convalescent lamb serum was as bactericidal as the whole serum. Absorption of convalescent serum with lipopolysaccharide from P. haemolytica abolished bactericidal activity, suggesting that this antigen may be a target for antibody in the bactericidal complex. PMID- 2967580 TI - T lymphocyte development in horses. I. Characterization of monoclonal antibodies identifying three stages of T lymphocyte differentiation. AB - Six monoclonal antibodies reacting with equine T lymphocytes at different stages of maturation were selected from antibodies produced against lymphoid cell preparations. EqT12 and EqT13 antibodies identified subsets of cortical thymocytes with high terminal deoxynucleotidyltransferase (TdT) activity and no phytolectin responsiveness. EqT12+ thymocytes were scattered throughout the cortex while EqT13+ thymocytes were located in the subcapsular cortex. EqT12 bound to small numbers of bone marrow cells, splenocytes, and circulating lymphoid cells, but not to mature T lymphocytes. EqT13 bound to very small numbers of bone marrow cells but not to more mature lymphocytes. EqT6 and EqT7 reacted with a large population of cortical thymocytes with high TdT activity and no phytolectin responsiveness. EqT2 and EqT3 bound primarily to medullary thymocytes with low TdT activity. Eq2+ thymocytes responded to phytolectin stimulation while EqT3+ thymocytes did not. EqT2 and EqT3 bound to 33% and 91% of circulating T lymphocytes, respectively. The T lymphocytes bound by both antibodies included cells capable of suppressing a mixed lymphocyte reaction. Thus, EqT12 and EqT13 identify cells with the functional characteristics of prothymocytes. EqT6 and EqT7 identify resident cortical thymocytes, and EqT2 and EqT3 identify a subpopulation of mature T lymphocytes and all mature T lymphocytes, respectively. PMID- 2967581 TI - [Biorhythm of the indicators of lipid metabolism and blood coagulation in patients with ischemic heart disease and its significance in the health resort practice]. PMID- 2967582 TI - [Value of physio-balneological factors in the preventive treatment of patients with ischemic heart disease in a sanatorium-preventorium]. PMID- 2967584 TI - [Use of the electronic-computer technic in health resort treatment and physiotherapy]. PMID- 2967583 TI - [Effectiveness of the treatment of patients in a sanatorium-dispensary]. PMID- 2967585 TI - [Efficacy of the cytologic diagnosis of malignant tumors of the thyroid gland (based on data from the Kuibyshev Oblast Oncologic Clinic)]. AB - The study deals with the possible scope of cytologic procedures in establishing the nature and histologic pattern of malignant tumors of the thyroid. Cytologic procedures proved effective in diagnosing A-cell follicular carcinoma (63.6%), A cell papillary carcinoma (91.7%), B-, C- and squamous cell cancer and lymphosarcoma (100%). PMID- 2967586 TI - [Status of the nervous system in cotton industry workers (weavers and spinners)]. PMID- 2967587 TI - [Laparoscopic diagnosis of infertility of tubal origin]. PMID- 2967588 TI - [Treatment of skin changes after extravenous administration of adriamycin]. PMID- 2967589 TI - [Parenteral contraceptive drugs: depot progestins]. AB - Depot progestins as injectables, implantables or vaginal rings are suitable for contraception in those female patients, in which risk factors (e.g. cardiovascular risk) exclude the use of estrogen-progestin mixtures. In this paper the mode of action, indications, contraindications, advantages and disadvantages of the various methods using depot-progestins are discussed. Injectables contain either medroxyprogesterone acetate or norethistronenantate; both steroids are released slowly within a limited time interval (2 to 4 months) out of a depot. The major effect is a change of the cervical mucus. Side-effects are disturbances of the menstrual cycle (e.g. breakthrough bleedings) as well as an amenorrhea after frequent use (up to 50% of all cases). The subdermal implantables (Norplant 2 or 5) release levonorgestrel out of a depot over a time period of at least 5 years. Steroid plasma levels are lower than in those patients using a progestin-only pill. Side-effects of implantables are disturbances of the menstrual cycle (e.g. breakthrough bleeding); in patients who desire to conceive a child or suffer from undesirable side-effects the implantables can be removed at every time. The progestin releasing vaginal rings are in a stage of controlled clinical trials. The advantages depend on ethe easy mode of administration (implantation or removal). Side-effects are also breakthrough bleedings. PMID- 2967590 TI - Possible nutrient mediators in psoriasis and seborrheic dermatitis. I. Prevalence, etiology, symptomatology, histological and biochemical features. PMID- 2967591 TI - Possible nutrient mediators in psoriasis and seborrheic dermatitis. II. Nutrient mediators: essential fatty acids; vitamins A, E and D; vitamins B1, B2, B6, niacin and biotin; vitamin C selenium; zinc; iron. PMID- 2967592 TI - Metabolism of amlodipine in the rat and the dog: a species difference. AB - 1. Following oral and i.v. doses of 14C-amlodipine to rat and dog, 40-50% of the dose was excreted in the urine indicating that the oral dose was well absorbed. Urinary and faecal excretion in rat was essentially complete within 48 h but was prolonged during 168 h in dog. 2. Metabolite patterns were dissimilar for rat and dog for both urine and faeces. The majority (about 95%) of the urinary metabolites were identified for both species; unchanged drug accounted for 10% and 2% of the urinary radioactivity in rat and dog respectively. 3. In rat, the principal route of metabolism involved cleavage of the 5-methoxy-carbonyl group of both the parent dihydropyridine and its pyridine analogue. In contrast, metabolism in dog involved oxidative deamination of the 2-aminoethoxy-methyl side chain. 4. Secondary metabolism in both rat and dog was similar to that of other calcium channel blockers of the dihydropyridine class, with oxidation to the pyridine form being followed by aliphatic hydroxylation in the 6-position or O dealkylation in the 2-position and lactonization. PMID- 2967593 TI - Metabolism and kinetics of amlodipine in man. AB - 1. The disposition of amlodipine, R,S,2-[(2-aminoethoxy)methyl]-4-(2 chlorophenyl)-3-ethoxycarbonyl- 5- methoxycarbonyl-6-methyl-1,4-dihydropyridine has been studied in two human volunteers using single oral and intravenous doses of 14C-amlodipine. The drug was well absorbed by the oral route while the mean oral bioavailability for unchanged drug was 62.5%. 2. Renal elimination was the major route of excretion with about 60% of the dosed radioactivity recovered in urine. Mean total recovered radioactivity in urine and faeces amounted to 84% for both the oral and intravenous routes. 3. Apart from a small amount of unchanged amlodipine (10% of urine 14C), only pyridine metabolites of amlodipine were excreted in urine. The majority (greater than 95%) of the metabolites excreted in the 0-72 h post-dose period were identified; the major metabolite was 2-([4-(2 chlorophenyl)-3-ethoxycarbonyl-5-methoxycarbonyl-6-methyl- 2-pyridyl]methoxy) acetic acid and this represented 33% of urinary radioactivity. The data indicate that oxidation of amlodipine to its pyridine analogue is the principal route of metabolism with subsequent metabolism by oxidative deamination, de-esterification and aliphatic hydroxylation. 4. For the two volunteers, amlodipine concentrations in plasma declined with a mean half-life of 33 h, while slower elimination of total drug-related material from plasma was observed, consistent with prolonged excretion (up to 12 days) of metabolites in urine and faeces. Only amlodipine and pyridine metabolites were found in the circulation. As these pyridine derivatives have minimal calcium antagonist activity the efficacy of amlodipine in man can most probably be attributed to the parent drug. PMID- 2967594 TI - [Stepwise study procedure of sugar substitutes--preliminary study with enzymes. 4. Glucosyltransferases of Streptococcus mutans AHT]. AB - A continuous procedure for the simultaneous enzymatic measurement of the release of free fructose and glucose was adapted to the kinetic conditions of the synthesis of polysaccharides from sucrose by glucosyltransferases from Streptococcus mutans AHT. Initial velocities, Km of sucrose, and the efficiency of the formation of glucans from sucrose can be determined. Longtime incubations with the isolation of soluble and insoluble glucans as an established method were compared with the new procedure. As examples of the effect of sugar substitutes on glucosyltransferases, data on leucrose, nystose, Palatinit, xylitol, leucritol and polyglucose PL-3 are presented. The results provide a preliminary assessment of sugar substitutes such as non-cariogenic sweeteners. PMID- 2967596 TI - [Keratosis palmoplantaris diffuse circumscripta (Thost-Unna) with micropenis]. AB - We report on a 2-year-old boy showing diffuse palmoplantar keratoderma (Thost Unna's disease) in association with congenital penis hypoplasia. Family history research proved palmoplantar keratoderma autosomally dominant over 5 generations. The boy's 6-month-old brother also shows penis hypoplasia, as yet, however, without signs of palmoplantar keratoderma. So far we are not able to settle the question whether this case represents a clinical entity of its own or merely a chance association of rare symptoms. PMID- 2967595 TI - [Side effects of external ophthalmologic drugs]. AB - 43 patients with suspected adverse reactions to ophthalmic medicaments were tested using three different methods: normal patch test on back skin (ENH), patch test on stripped back skin (HSA) and on scarified skin of the forearm (SKT). The ENH was positive in only 9 patients (20.0%) with one of the used preparations whereas on stripped skin positive reactions were observed in another 14 patients (32.6%). The SKT was solely positive in 10 patients (23.7%). Clinical relevance of these test results has been ascertained in most cases. However, reactions on stripped skin as well as on scarified skin may be false positive. The antibiotics neomycin and gentamicin were the major allergens (13.9% and 9.3%). Some ophthalmic medicaments produced rather severe irritant reactions on scarified skin, confirmed by a positive conjunctival exposure test. In order to detect weak sensitizations or cumulative irritant reactions in patients with long term use of eye medicaments, the application of a method with increased sensitivity such as the patch test on stripped skin or the scarification technique is recommended. PMID- 2967597 TI - [Strategic considerations in the diagnosis and therapy of problem patients with back surgery]. PMID- 2967598 TI - [Measurement of microvascular perfusion by laser Doppler flowmetry. Some uses in clinical practice]. PMID- 2967599 TI - [Comparison of laser Doppler flowmetry and plethysmography]. PMID- 2967600 TI - [Incidence and reasons for late consultations of patients with skin cancer]. PMID- 2967602 TI - [Dental-surgical procedures in hematological disorders]. PMID- 2967601 TI - [Various aspects of the incidence of acne among adolescents]. PMID- 2967603 TI - [Infection control measures in dentistry]. PMID- 2967604 TI - [The handicapped in the dental practice]. PMID- 2967605 TI - [Temporary denture fixation. Retention of the tooth root for temporary fixation of partial and hybrid dentures]. PMID- 2967607 TI - [Experiences during treatment with ether]. PMID- 2967606 TI - [Is the use of low palladium and non-precious metal alloys worthwhile?]. PMID- 2967609 TI - [The Optec-HSP System]. PMID- 2967608 TI - [Probond. A precious metal conserving reconstruction method for metal-ceramics]. PMID- 2967610 TI - [Metal-ceramics]. PMID- 2967611 TI - [Composites as adhesive cements? Initial clinical experiences in crown and bridge prosthetics]. PMID- 2967612 TI - [Transcranial Doppler sonography in healthy persons]. AB - The authors describe the technical parameters and methodology of transcranial Doppler sonography as applied to studying the blood flow in vessels of the brain base. The results of the examination carried out in 25 healthy subjects have shown a high informative value of transcranial Doppler sonography in studying the circulation in the arterial and venous collectors. The new method is compared with the routine Doppler sonography and echopulsography. PMID- 2967613 TI - Measurements of common femoral artery flow velocity in the evaluation of aortoiliac atherosclerosis. Comparisons between pulsatility index, pressures measurements and pulse-volume recordings. AB - Blood flow velocity in the common femoral artery was measured in 52 limbs of 45 patients with radiologic signs of aortoiliac atherosclerosis. Group A limbs had significant obstruction of the aortoilac segment; group B had not. 'Significant' implied a transobstruction pressure gradient of greater than or equal to 10 mmHg at rest or greater than or equal to 20 mmHg after intra-arterial injection of papaverine. Pulse-volume recording (PVR) and non-invasive measurement of the common femoral artery pressure (FAP) were also performed. Pulsatility index (PI) less than 3.9 and PVR amplitude less than 13 mm indicated significant obstruction, whereas PI greater than 5.4 and PVR amplitude greater than 20 mm were observed only in limbs without significant pressure gradient across the aortoiliac segment. PI 3.9-5.4 and PVR amplitude 13-20 mm were found in both groups. FAP and flow velocity during reactive hyperemia did not adequately distinguish the two groups, because of value overlap. In assessing the hemodynamic significance of aortoiliac obstruction, PI and PVR in the common femoral artery may be useful supplements to clinical examination, but measurement of the pressure gradient across the aortoiliac obstruction before and after vasodilation probably is most reliable. PMID- 2967614 TI - Assessment of gastrointestinal blood flow with laser Doppler flowmetry in patients with chronic radiation injury. AB - Laser Doppler flowmetry was used for pre- and intraoperative assessment of gastrointestinal blood flow and vascular damage in eight patients with late radiation injury of the gastrointestinal tract. The flowmeter recordings were compared with reference values previously obtained in unaffected tissue from corresponding gastrointestinal sites. Marked reduction of blood flow was demonstrated in macroscopic areas of radiation injury in all eight patients. Five also showed varying degrees of disturbed blood flow in adjacent macroscopically normal bowel segments with histologically varying severity of radiation injury. In a ninth patient radiation injury was clinically suspected but could not be demonstrated with laser Doppler flowmetry or histologically. The extent of radiation injury often is underestimated from clinical signs, and laser Doppler flowmetry during surgical treatment of late radiation injury to the gastrointestinal tract provides substantial information concerning the extent of vascular damage. PMID- 2967616 TI - Postoperative care: antagonism of drugs used in anaesthesia: muscle relaxants. PMID- 2967615 TI - New observations on the significance of nipplelike protrusions in the nuclei of endocervical cells. AB - Nuclear nippling was noted in the endocervical columnar cells in smears taken from women of child-bearing age who were using a progesterone-only contraceptive. The phenomenon occurred most consistently in smears from women who were amenorrheic, a condition indicative of a marked decrease of estrogen activity. These observations do not support the previous explanations linking nuclear nippling to estrogenic stimulation. Close observation of the progress of these nuclear protrusions showed that they represented an arrested attempt at nuclear division, with the inhibition possibly due to the rapid rise of the progestogen level, which may be caused either by the use of progesterone-only contraceptives or by the onset of ovulation. The presence of nuclear nippling in normal smears may thus establish the occurrence of ovulation; its observation in metastatic tumors should indicate that the primary tumor has a columnar cell component. PMID- 2967617 TI - Solubility of neurofibrillary tangles and ultrastructure of paired helical filaments in sodium dodecylsulphate. AB - Temporal cortex from 14 cases of Alzheimer-type dementia and 6 cases of Down's syndrome, all selected for severe Alzheimer pathology, was homogenised in distilled water, NaOH, or sodium dodecylsulphate (SDS) containing 0.1% beta mercaptoethanol. The homogenates were stained with Congo red, and the neurofibrillary tangles and plaque cores were counted under crossed-polarisation microscopy. The number of tangles and plaque cores in the water-treated extracts was not related to age, sex, post-mortem interval or duration of dementia. The number of tangles after extraction in SDS or NaOH, as a percentage of tangles in water-treated extracts, was 57 +/- 25 (mean +/- SD) for 1% SDS, 43 +/- 17 for 5% SDS and 37 +/- 22 for 0.2 M NaOH. Plaque cores were essentially insoluble in all three agents. The percentage of tangles insoluble in 1% SDS did not correlate with age or post-mortem interval but decreased with increasing duration of dementia. Enhanced tangle solubility with increasing duration of dementia suggests that the nature of tangles changes with time; one possibility is that this reflects transformation of intracellular to extracellular tangles. Paired helical filament (PHF) length and the number of repeats per PHF were measured in electron micrographs of PHF prepared with and without treatment by 1% SDS. There was no significant multimodality of PHF length to suggest that PHF broke at regular intervals. The mean repeat length (PHF length/number of repeats) was greater for PHF isolated in the presence of 1% SDS than in its absence, showing that SDS affects ultrastructure by untwisting PHF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967618 TI - Localisation of calcium-activated adenosine-triphosphatase (Ca2+-ATPase) in intracerebral arterioles in acute hypertension. AB - The plasma membrane calcium-activated adenosine triphosphatase (Ca2+-ATPase) is known to regulate intracellular calcium levels. This enzyme was localised in intracerebral cortical vessels of normotensive and acutely hypertensive rats. Of interest was whether the arterioles that develop increased permeability to horseradish peroxidase (HRP) in acute hypertension demonstrate any alteration in localisation of Ca2+-ATPase as compared to normotensive controls. Rats were injected with HRP intravenously and acute hypertension was induced by a 2-min infusion of angiotensin amide. Following perfusion of fixative, brains were sliced and reacted for demonstration of HRP reaction product and Ca2+-ATPase. Normotensive rats showed discontinuous distribution of Ca2+-ATPase on the outer plasma membranes of endothelial, smooth muscle and adventitial cells of arterioles. The localisation of Ca2+-ATPase in pinocytotic vesicles present in endothelial and smooth muscle cells was quite striking. Focal cortical areas of hypertensive rats showed increased arteriolar permeability to HRP. Permeable arterioles showed marked reduction of Ca2+-ATPase on the outer plasma membranes of endothelium and smooth muscle cells as compared to nonpermeable arterioles of the same animals and arterioles of normotensive controls. The latter finding suggests that calcium may be involved in increased cerebrovascular permeability mechanisms in acute hypertension. PMID- 2967619 TI - [Effects of nicotinamide on sinoatrial conduction time and sinus node recovery time in rabbit hearts]. PMID- 2967620 TI - [Photodynamic action of two hematoporphyrin derivatives on transport and intracellular metabolism of adenosine in hepatoma cells]. PMID- 2967621 TI - Osteoporosis and back pain among the elderly. AB - Questionnaire responses from 120 men and 337 women over the age of 50 years were studied to determine the prevalence of back pain among the elderly. In order to gain a rough indication of the back pain among elderly women which might be due to osteoporosis, the prevalence was compared in the two sexes. The prevalence of back pain without radiation to the legs and concomitant morbidity was found to be similar among men and women up to the 70-79-year age-group. After this age the prevalence was higher in women. Those with exceptional loss of body height or kyphosis had a high prevalence of back pain, while those who had sustained previous hip or radius fractures did not. There was increasing prevalence of back pain among women with increasing number of previous fractures. The study gives little indication of serious morbidity of osteoporosis in the form of back pain before very old age. PMID- 2967622 TI - [Endocrine therapy of prostatic carcinoma with slow release (depot) formulation of the LH-RH analog ICI 118630 (Zoladex)]. AB - To investigate the clinical efficacy, safety and endocrinology of ICI 118630 (Zoladex) depot formulation at 3 different dose levels (0.9, 1.8 and 3.6 mg), 90 patients were randomized to receive either one of the 3 doses from April, 1985 to March, 1986 in 28 centers. The depot preparation was injected subcutaneously every 4 weeks 3 times (for up to 12 weeks). Clinical efficacy was evaluated in terms of tumor response and overall subjective response. In 70 patients eligible for tumor response evaluation, 14 out of 22 (63.6%) in the 0.9 mg group, 11 out of 23 (47.8%) in the 1.8 mg group, and 17 out of 25 (68.0%) in the 3.6 mg group showed clinical improvement, that is, either complete response or partial response. In 72 eligible patients for overall subjective response evaluation, clinical subjective improvement was observed in 75.0, 81.8 and 88.0% of the patients in the 3 groups, respectively. There was no significant difference between the groups. As for endocrinology, there were 75 eligible patients. Endocrinological effect was observed in 23 out of 25 (92.0%) in the 0.9 mg group, 100% in both 1.8 mg and 3.6 mg groups. There was no significant difference between the groups. Castration was achieved by week 3.5 +/- 1.7 of therapy on average and by week 2 in the earliest case. There was no significant difference in incidence of side effects between the 3 groups: 5 out of 26 (19.2%) in the 0.9 mg group, 8 out of 29 (27.6%) in the 1.8 mg group, and 2 out of 30 (6.7%) in the 3.6 mg group. Flares presented as an increase in bone pain in 2 and as ureteric obstruction in 2 all in the 1.8 mg group but none in the other 2 dose groups. These flares disappeared on further treatment with Zoladex. These patients showed a clinical-response. The blood level of Zoladex was dose dependent, reaching its peak at week 2 of therapy in all 3 dose groups. There was no evidence of accumulation. Since these results demonstrate that 3.6 mg produces medical castration earlier, it may well be considered as an optimal dose in men. PMID- 2967623 TI - Improvement in regional wall motion after percutaneous transluminal coronary angioplasty during acute myocardial infarction: utility of two-dimensional echocardiography. AB - In the setting of acute myocardial infarction, 16 patients undergoing successful coronary angioplasty (PTCA) within 6 hours of presentation (group I) and eight patients receiving conventional medical therapy (group II) were studied by serial two-dimensional (2D) echocardiography to assess the functional recovery of myocardium. All patients underwent 2D echocardiograms within 24 hours of presentation and at a minimum of 6 days after admission. Wall motion analysis was quantified with a wall motion score index based on 16 left ventricular wall segments. Wall motion score index improved significantly from early to late echocardiographic study in the patients undergoing PTCA (1.65 +/- 0.29 to 1.40 +/ 0.30; p less than 0.001), whereas the index did not improve in the conventionally treated group (1.54 +/- 0.26 to 1.58 +/- 0.25; p = NS). One patient in group II had a greater than or equal to 10% improvement in wall motion score index compared to 11 of 16 in group I (p less than 0.01). In all cases improvement in wall motion score index was due to improvement in regional wall motion in the area of infarction. In group I, 40 of 77 (52%) infarct zone segments showed improvement of at least one grade, versus 4 of 28 (14%) segments in group II (p less than 0.001). These data indicate that regional myocardial function improves in the majority of patients undergoing successful PTCA as emergency therapy for acute myocardial infarction and that serial 2D echocardiography is an excellent means to quantify this improvement. PMID- 2967624 TI - Preservation of left ventricular ejection fraction during percutaneous transluminal coronary angioplasty by distal transcatheter coronary perfusion of oxygenated Fluosol DA 20%. AB - The cardioprotective efficacy of coronary perfusion during angioplasty was evaluated. Forty-two patients underwent transcatheter infusion of oxygenated Fluosol DA, 20% emulsion (FDA-20), a perfluorocarbon oxygen transport fluid, into the distal coronary artery during balloon inflations. Left ventricular function was continuously monitored by two-dimensional echocardiography, and left ventricular ejection fraction was quantitatively analyzed from the video record by an area-length method with a validated computer algorithm. Each patient had multiple nonperfused and perfused balloon inflations lasting more than 45 seconds. Nineteen of the 42 patients also received control solutions of oxygenated Ringer's lactate and nonoxygenated FDA-20. The ejection fraction of nonperfused sequences fell from a baseline value of 57 +/- 15% to 36 +/- 14% at 45 seconds of inflation time (p less than 0.0005). Falls of similar magnitude were seen in the lactated Ringer's and nonoxygenated FDA-20 perfused balloon inflations. The ejection fraction fall was associated with a 54% rise in end systolic volume (p less than 0.0005) and a 4% rise in end-diastolic volume (p = ns) compared to baseline. Inflations perfused with oxygenated FDA-20 showed a 45 second, left ventricular ejection fraction of 53 +/- 13% (p = ns compared to baseline), which was significantly greater (p less than 0.0001) than the 45 second ejection fraction of the nonperfused, or control solution perfused sequences. Results indicate that the profound fall in ejection fraction occurring during percutaneous transluminal coronary angioplasty can be ameliorated by distal coronary perfusion with an oxygenated perfluorocarbon emulsion. PMID- 2967625 TI - Effects of nitroglycerin and nifedipine on coronary and systemic hemodynamics during transient coronary artery occlusion. AB - Nitroglycerin (NTG) and nifedipine (NIF) have the potential to augment coronary blood flow in addition to reducing peripheral determinants of myocardial oxygen demand as a synergistic protective mechanism during ischemia. To examine these effects, systemic and coronary hemodynamic responses were measured continuously before and during brief periods of myocardial ischemia induced by left anterior descending coronary balloon occlusion in 26 patients undergoing angioplasty (PTCA). Data were compared for two matched occlusion periods, one control and one "drug" occlusion. In 17 patients (NTG group), 200 micrograms of intracoronary NTG was given immediately before coronary occlusion. In nine patients (NIF group), 10 mg of sublingual NIF was given 15 minutes before the "drug" occlusion. NTG significantly but transiently reduced mean arterial pressure (91 +/- 11 to 82 +/- 15 mm Hg, p less than 0.05) and augmented basal coronary blood flow (95 +/- 38 to 127 +/- 54 ml/min, p less than 0.05) but did not alter great vein blood flow (59 +/- 29 vs 61 +/- 29 ml/min) or coronary occlusion pressure (25 +/- 7 to 24 +/- 7 mm Hg) during ischemia. NIF significantly reduced systolic, diastolic, and mean arterial pressure (119 +/- 21 to 95 +/- 8 mm Hg, p less than 0.001) and heart rate-pressure product from control. NIF maintained basal great vein blood flow (125 +/- 41 to 106 +/- 57 ml/min) during reduced myocardial oxygen demand, but did not affect great vein blood flow (73 +/- 29 to 79 +/- 37 ml/min) or coronary occlusion pressures during ischemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967626 TI - Effects of atrial natriuretic factor on coronary hemodynamics and myocardial energetics in patients with heart failure. AB - Synthetic analogues of atrial natriuretic factor (ANF) have been developed for potential use as therapeutic agents in the treatment of congestive heart failure and hypertension. We studied the effects of 14 intravenous infusions of synthetic ANF (anaritide, human ANF 102-126) on coronary hemodynamics and myocardial energetics in six patients with heart failure. ANF infusion caused no change in coronary blood flow and a fall in coronary vascular resistance from 1.22 +/- 0.22 to 1.08 +/- 0.18 mm Hg-min/ml (p less than 0.05). Myocardial oxygen and lactate consumption were unchanged from baseline values. Mean arterial pressure fell from 91 +/- 4 to 78 +/- 3 mm Hg (p less than 0.01), right atrial pressure fell from 10 +/- 1 to 8 +/- 1 mm Hg (p less than 0.01), pulmonary capillary wedge pressure fell from 21 +/- 3 to 16 +/- 2 mm Hg (p less than 0.01), heart rate and cardiac index were unchanged, and systemic vascular resistance fell from 1346 +/- 130 to 1087 +/- 98 dyne-sec/cm5 (p less than 0.05). We conclude that infusion of ANF in hemodynamically effective doses in patients with heart failure decreases coronary vascular resistance with no change in coronary blood flow or myocardial oxygen or lactate metabolism. PMID- 2967628 TI - Transient loss of R wave during percutaneous transluminal coronary angioplasty. PMID- 2967627 TI - Superior vena cava syndrome treated by percutaneous transluminal balloon angioplasty. PMID- 2967629 TI - Atrial natriuretic peptide in AMI. PMID- 2967630 TI - A search for discomfort-inducing factors in carbonless copying paper. AB - Connections between various types of carbonless copying paper and the occurrence of work-related symptoms of the skin and mucous membranes were studied. The analysis started from the exposure factor, i.e., from the carbonless copying papers and not from the exposed persons. In total, 276 paper samples were investigated. One hundred ninety samples were carbonless copying papers for which information was obtained concerning both the handling of the papers and the symptoms occurring among those handling the papers. To distinguish between carbonless copying papers of different makes, gas chromatography of paper extracts was performed, sometimes combined with thin-layer chromatography of the color formers. By coding the paper samples, the analyses of the papers and the interviews of the exposed persons could be performed "blindly." In the carbonless copying paper most frequently associated with complaints, mono-iso-propylbiphenyl (MIPB) was used as solvent for the color formers. It is probable, however, that the symptoms connected to this paper were induced by some component of the paper other than MIPB. A covariation between papers associated with skin symptoms and mucous membrane symptoms was observed. Exposure to carbonless copying papers treated with desensitizing ink (D-ink) covariated statistically with work-related skin irritation. The causative factor was probably two specific D-inks available on the Swedish market at the period of the investigation. PMID- 2967631 TI - The philosophy of occupational safety and health regulation. PMID- 2967632 TI - Results of percutaneous transluminal coronary angioplasty by multiple, relatively low frequency operators: 1986-1987 experience. AB - The initial results of percutaneous transluminal coronary angioplasty (PTCA) in this institution were described previously and were comparable with national statistics despite the comparatively small number of cases performed by individual physicians. Two years later, the number and complexity of cases have increased significantly. In the present study, the experience of 3 physicians (group 1) who performed greater than 100 PTCAs (143 +/- 35, mean +/- standard deviation) was compared with that of 14 physicians (group 2) who performed less than 100 PTCAs (25 +/- 16) during a recent 12-month period. Group 1 and group 2 performed 430 and 351 PTCAs, with a 91 and 84% success rate, respectively (p less than 0.01). Group 1 and group 2 attempted dilatation of 595 and 444 narrowings with an 85 and 81% success rate, respectively (p less than 0.025). The differences reflect outcomes with "complex lesions," with which group 1 had a higher success rate than group 2 (81 vs 69%, p less than 0.05) and a lower complication rate (3.1 vs 7.5%, difference not significant). Outcomes with "simple lesions" were similar for the 2 groups (93 vs 90%). As a result, the conclusions of the previous study should be modified. The present data suggest that while low frequency operators can perform PTCA of "simple lesions" with quite satisfactory results in the setting of an institution in which large numbers of PTCAs are performed and in which an expert team is available for support, outcomes with "complex lesions" are likely to be better in more experienced hands. PMID- 2967633 TI - Stability of multilead ST-segment "fingerprints" over time after percutaneous transluminal coronary angioplasty and its usefulness in detecting reocclusion. AB - Multilead ST-segment recordings taken during percutaneous transluminal coronary angioplasty (PTCA) could function as an individualized noninvasive template or "fingerprint," useful in evaluating transient ischemic episodes after leaving the catheterization laboratory. To evaluate the reproducibility of such ST-segment patterns over time, these changes were analyzed in patients grouped according to the time between occlusion and reocclusion. For the patients in group 1, the study required comparing their "fingerprints" in repeat balloon inflation during PTCA (reocclusion in less than 1 hour), for those in group 2, comparing ST "fingerprints" during PTCA with ST changes during spontaneous early myocardial infarction (reocclusion in 24 hours) and in group 3, comparing ST "fingerprints" with ST changes during repeat PTCA for restenosis greater than 1 month after the initial PTCA. The ST "fingerprints" among the 20 patients in group 1 were identical in 14 cases (70%) and clearly related in another 4 (20%). Of the 23 patients in group 2, 12 (52%) had the same and 8 (35%) had related patterns. Of 19 patients in group 3, 8 (42% had the same pattern and 8 (42%) had related patterns. Thus, ST fingerprints were the same or clearly related with reocclusion in the same patient from less than 1 hour to greater than 1 month after initial occlusion in 87% of patients overall, in 90% in less than 1 hour, in 87% in less than 24 hours and in 84% greater than 1 month later. Multilead pattern ST-segment "fingerprints" may serve as a noninvasive marker for detecting site-specific reocclusion. PMID- 2967634 TI - Results of percutaneous transluminal coronary angioplasty for angina pectoris early after acute myocardial infarction. AB - Between May 1980 and July 1985, 70 patients underwent percutaneous transluminal coronary angioplasty (PTCA) for angina occurring 24 hours after and within 30 days of acute myocardial infarction (32 with Q-wave infarction and 38 with non-Q wave infarction). One-vessel disease was present in 42 (60%) and multivessel in 28 (40%); the mean ejection fraction was 0.56 (greater than or equal to 0.50 in 77% of patients). PTCA was successful in 56 patients (80%) and after introduction of steerable dilating systems in February 1983 this rate became 86%. The success rate for complete occlusions was 76%. The interval from myocardial infarction to PTCA was similar in patients with successful dilation (12.7 +/- 8.1 days) and those without (13.4 +/- 8.0 days). PTCA failed in 14 patients (20%); 8 underwent emergency coronary artery bypass for acute occlusion and 4 of 6 patients whose lesions could not be crossed had elective bypass surgery. There was 1 operative death. No patient sustained a Q-wave infarction. Three patients had non-Q-wave infarctions after technically successful PTCAs. Mean follow-up was 27 months (6 to 67 months). Of the 56 patients successfully dilated, 14 (25%) had 15 cardiac events during follow-up: death (1), non-Q-wave infarction (2), repeat PTCA (7), coronary bypass (4) and recurrence of severe angina (1). The cumulative mortality was 3% and the reinfarction rate was 7% (no Q-wave reinfarctions). Forty-two (60%) of the 70 patients were free of complicating events acutely and during follow-up.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967635 TI - Percutaneous transluminal coronary angioplasty in patients with two or more previous coronary artery bypass grafting operations. AB - Between 1979 and 1986, 65 of 76 patients (86%) (82% men, with a mean age of 58 +/ 8 years) with greater than or equal to 2 previous coronary artery bypass grafting (CABG) operations and symptomatic myocardial ischemia underwent successful percutaneous transluminal coronary angioplasty (PTCA). Sixty-two patients had 2 prior CABG operations, 10 had 3 and 4 had 4. Clinical characteristics included prior myocardial infarctions in 49 (65%), severe angina (class III or IV) in 47 (62%) and left ventricular ejection fraction less than or equal to 35% in 13 (17%). There were 139 lesions dilated: 1 lesion in 39 (51%), 2 in 22 (29%) and greater than or equal to 3 in 15 (20%) patients. Arterial lesions were successfully dilated in 71 of 81 cases (88%), vein grafts in 44 of 53 (83%) and mammary artery grafts in 3 of 5 (60%). In 12 patients, PTCA was used to dilate significant lesions less than 15 days after CABG in vessels which were unable to be bypassed. Significant complications were encountered in 4 patients (5%). These included 3 of 53 vein graft dilatations with embolization (6%), with 1 resulting in infarction and death, and 1 patient dying after emergency CABG. At hospital discharge, 65 patients were clinically improved. An apparent symptom related lesion recurrence occurred in 23 of 65 patients (35%), with 5 patients dying of cardiac causes, 4 having CABG without previous angiography and 12 of 14 patients undergoing repeat successful PTCA (mean time and standard deviation 9 +/ 6 months).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967636 TI - Atrial natriuretic peptide during supraventricular tachycardia and relation to hemodynamic changes and renal function. AB - Changes in plasma levels of atrial natriuretic peptide (ANP) and arginine vasopressin were studied in 5 patients during and after a 30-minute period of induced supraventricular tachycardia (SVT). Immediately after the induction of SVT, plasma ANP levels began to increase, peaked at 32 minutes (+734% increase on average) and then gradually decreased. The mean plasma arginine vasopressin levels decreased during SVT, but the differences were not significant. When plasma ANP levels during SVT were compared with the simultaneously measured hemodynamic variables, a significant positive correlation (r = 0.73, p less than 0.001) was observed between plasma ANP levels and pulmonary capillary wedge pressure. Induced SVT was associated with increased urinary sodium and potassium excretion, increased urine flow and increased free water clearance. Concomitantly, glomerular filtration rate significantly increased (+77%) with an increase in filtration fraction. Although no significant change was observed in plasma renin activity, plasma aldosterone concentrations decreased during and after SVT. These results suggest that increased left atrial pressure stimulates ANP release during SVT and that increased glomerular filtration rate and decreased aldosterone secretion by ANP, in addition to the inhibition of water reabsorption by decreased arginine vasopressin, may be responsible for natriuresis and diuresis associated with SVT. PMID- 2967637 TI - The enigma of insulin resistance and hypertension. PMID- 2967638 TI - Left ventricular dysfunction due to chronic right ventricular pressure overload. PMID- 2967639 TI - Corneal fistulas and their management. AB - We reviewed three representative cases of chronic corneal fistula formation and provide a systematic approach to the assessment and therapeutic alternatives for this problem. In two of our patients, the fistula was managed surgically. The third patient developed endophthalmitis, which resulted in loss of light perception. Chronic corneal fistulization is a rare clinical entity resulting from malapposition of corneal tissue after traumatic, surgical, or infectious perforation. Fistulas may result in prolonged or recurrent hypotony, peripheral anterior synechia formation, or endophthalmitis. Accurate assessment of the risks associated with corneal fistula formation takes into account the type of fistula, its location in the cornea, and the condition of the ocular adnexae. We reviewed the risk factors that will determine the urgency and type of therapy used. PMID- 2967640 TI - Paradoxical organ-specific adaptations to streptozotocin diabetes mellitus in adult rats. AB - Adult male Fisher rats injected with streptozotocin (Stz) to produce diabetes mellitus demonstrated a significant loss of total body weight associated with adipose and muscle tissue wasting. Paradoxically, intestinal mass and length were increased in Stz-treated rats despite catabolism of other tissues. Concomitant with increased intestinal mass, food and water intake increased significantly in Stz-diabetic animals. Renal weight was not reduced despite the fall in total body weight. It is proposed that the adult Stz-diabetic rat responds to a loss of available insulin by polyphagia, polydipsia, and catabolism of adipose and muscle tissue and that a large percentage of available synthetic fuel is devoted to the production of additional intestinal tissue. PMID- 2967641 TI - Aldosterone and PCO2 enhance rubidium absorption in rat distal colon. AB - Recent studies of rabbit colon have indicated the presence of a vanadate sensitive K+-dependent proton pump, suggesting the existence of an H+-K+-ATPase. The participation of such a mechanism for colonic K+ absorption in the rat has not been determined. To this purpose, we attempted to detect the presence of pH linked mechanisms for K+ absorption in rat distal colon using 86Rb as a marker for K+. We found that Rb+ absorption in Na-Ringer directly correlated with the in vitro partial pressure of CO2 (PCO2) in aldosterone-stimulated but not in control rats. Similar studies performed using Na-free Ringer demonstrated that PCO2 markedly augmented Rb+ absorption in both control and aldosterone-stimulated rat colon. Rb+ absorption was inhibited by orthovanadate, SCH28080, and mucosal ouabain in Na-free Ringer, but there was no effect of omeprazole, furosemide, or bumetanide. Barium applied to the serosa was also effective in inhibiting Rb+ absorption, suggesting that Rb+ exit from the cell was conductive. These findings are consistent with the presence of an active K+ pump that is activated by pretreatment with aldosterone and increased in vitro PCO2 and that is inhibited by orthovanadate, SCH28080, and mucosal ouabain. The constellation of findings suggests that participation of an ATPase that is not typical of either Na+-K+ ATPase or H+-K+-ATPase. PMID- 2967643 TI - [Therapy of endocrine disorders in abortion]. PMID- 2967642 TI - Biologic fixation of ligament prostheses and augmentations. An evaluation of bone ingrowth in the dog. AB - The biologic fixation (bone ingrowth) of three prosthetic ligament devices (bovine xenograft Xenotech Laboratories, Inc., Irvine, CA; Gore-Tex, W. L. Gore and Associates, Flagstaff, AZ; and knitted dacron, Stryker, Kalamazoo, MI) and one ligament augmentation device (LAD, braided polypropylene, 3M, St. Paul, MN) were evaluated in vivo. The devices were placed unstressed, in an extraarticular location in the cortical-cancellous bone of the proximal femur and humerus of dogs. Six months following implantation tissue ingrowth was evaluated using high resolution radiography, routine histology, scanning electron microscopy, and biomechanical tensile tests to measure the ultimate (maximum) pull-out strength of the devices. The knitted dacron and Gore-Tex prosthesis showed marked ingrowth of trabecular bone into the interstices of the device with evidence of bone growth around individual fibers of the prosthesis. The bovine xenograft demonstrated bone immediately adjacent to the material with evidence of spicular ingrowth of bone into the natural clefts of the prosthesis. The LAD showed no evidence of bone ingrowth and demonstrated a fibrous tissue interface between the prosthesis and surrounding trabecular bone. Six months following implantation the pull-out strengths of the devices were as follows: knitted dacron, 445.7 +/- 151.0N; Gore-Tex, 438.1 +/- 131.94N; bovine xenograft, 332.8 +/- 64.2N; and LAD, 78.4 +/- 47.9N. Statistical analysis revealed no significant difference between the pull-out strengths of the knitted dacron, Gore-Tex, and bovine xenograft.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967644 TI - Mivacurium chloride (BW B1090U)-induced neuromuscular blockade during nitrous oxide-isoflurane and nitrous oxide-narcotic anesthesia in adult surgical patients. AB - The neuromuscular and cardiovascular effects of mivacurium were studied in 90 adult patients during nitrous oxide-oxygen-isoflurane (n = 45, ISO group) and nitrous oxide-oxygen-narcotic (n = 45, BAL group) anesthesia. Neuromuscular blockade was measured using electromyographic activity of the adductor pollicis muscle after supramaximal stimulation of the ulnar nerve at 2 Hz for 2 seconds at 10-second intervals. To estimate dose-response relations, three subgroups of nine patients in the ISO group received mivacurium doses of 0.025, 0.03, and 0.04 mg/kg, respectively. Similarly, three subgroups of nine patients in the BAL group received mivacurium doses of 0.03, 0.04, and 0.05 mg/kg, respectively. The ED50 and ED95 of mivacurium in each group were estimated from linear regression plots of log dose vs probit of maximum percentage depression of neuromuscular function. The estimated ED50 values for the ISO and BAL groups were 0.029 and 0.041 mg/kg, respectively. The estimated ED95 values for the ISO and BAL groups were 0.045 and 0.058 mg/kg, respectively. Recovery indexes were measured in 26 patients who received ED95 or greater doses of mivacurium in either the ISO or BAL groups. The recovery index was shorter in the BAL group (5.5 +/- 1.6 minutes [n = 10]), than in the ISO group (7.4 +/- 3.0 minutes [n = 16]). The addition of isoflurane (0.5 0.75% end-tidal concentration) to nitrous oxide-narcotic anesthesia augments the degree of neuromuscular blockade from a given dose of mivacurium and also prolongs the recovery index. PMID- 2967647 TI - [Patterns of stomatological care in questionnaire responses of workers at 4 industrial plants]. PMID- 2967646 TI - Analysis of a horse family with a crossing-over between the ELA complex and the A blood group system. AB - A horse family in which a recombination occurred in the chromosome region coding for the serological specificities of the ELA complex and those of the A blood group system of a mare was further analysed by mixed lymphocyte reaction (MLR) and Southern blot hybridization. This family consisted of a stallion, a mare and five full sibs. The stallion and the mare were heterozygous for internationally recognized ELA specificities while only the mare was heterozygous for the A blood group system. MLR between all members of the family confirmed that the stallion possessed two different ELA haplotypes and suggested that recombination in the mare occurred outside the segment delimited by the ELA-A locus and the MLR region. DNA samples from all individuals were investigated by Southern blot analysis using three restriction enzymes (EcoRI, HindIII or TaqI), three human HLA probes (one of class I cDNA and two of class II probes), one cDNA (DR beta) and one genomic (DQ alpha). Class I and class II restriction fragments of the mare segregated in accordance to the ELA specificities and thus clearly confirming that the crossing-over did not occur between the ELA-A gene and the class I, class II region nor between DR beta and DQ alpha subsets. The A blood group genetic determinants would thus be situated outside the ELA region defined by class I and class II genes. PMID- 2967645 TI - Long-term beneficial effects of PTCA on segmental early relaxation in disease of the left anterior descending coronary artery. AB - The relationship between regional left ventricular (LV) motion and global pressure relaxation of the left ventricle remains unclear. To clarify the recent concept of segmental early relaxation in coronary artery disease, the authors investigated two groups of patients. In group I, all 12 patients (mean age 47 +/- 7 years) exhibited evidence of a normal heart after an extensive investigation. In group II, 25 patients (55 +/- 7 years) presented an isolated stenosis of the left anterior descending coronary artery, and they underwent a hemodynamic investigation before and after (six to nine months) a durable successful percutaneous transluminal coronary angioplasty (PTCA). After all conventional hemodynamic measurements had been done, a quantitative frame-by-frame analysis of left ventricular wall motion was conducted. The authors' method is derived from that of Ingels, applying to LV cineangiograms filmed in 30 degrees right anterior oblique view at a 50 frames/second rate. Thus segmental wall motion is analyzed in terms of amplitudes (%), velocities of shortening and lengthening in circumferences/second (circ/sec), and times of events (%). Statistical results took into account the reproducibility of the method. Main results regarding the control state of group II consisted of an asynergic motion of the anterior region taking place from end systole to early diastole: 1. Early end of contraction in anterior segments (% of systolic time interval: 88 +/- 14% vs 96 +/- 6% in group I, p less than 0.001) 2. Asynchronism at end systole (maximal velocity of shortening - 0.4 +/- 2.3 circ/sec in anterior segments vs 0.05 +/- 1.9 in inferior segments, p less than 0.02) 3. An early but poor outward anterior wall motion (anterior lengthening at 0.04 sec after the end of ejection 2.9 +/- 10% in group II versus 5.4 +/- 7.2% in group I, p less than 0.05) These abnormalities are strongly correlated with a significant impairment of peak negative diastolic pressure/diastolic time (dP/dt) (1500 +/- 400 mmHg. sec-1 vs 1850 +/- 410 in group I, p less than 0.02). Long-term beneficial effects of PTCA in group II were characterized by an almost complete normalization, both asynergy and relaxation taking place back within the normal range. The authors conclude that in this kind of patient, peak negative dP/dt could be an index of an asynergic segmental motion, this one being correctly analyzed and quantified on LV cineangiograms with our method. PMID- 2967649 TI - [Xanthine compounds with anti-arteriosclerotic effect in rabbits]. PMID- 2967648 TI - [Fluoride levels in bacterial plaque and saliva and the condition of periodontium in children living in regions with fluoridated and non-fluoridated water supply]. PMID- 2967650 TI - [Use of echocardiographic examinations for the evaluation of anatomical and functional state of the left ventricle in patients with arterial hypertension]. PMID- 2967651 TI - [Retrospective evaluation of prognostic factors in patients with aplastic anemia]. PMID- 2967652 TI - [Studies of the mitral leaflet prolapse syndrome with special reference to angiographic examination]. PMID- 2967653 TI - [Effect of beta-adrenergic blockade on the adaptation of the heart to increased arterial blood pressure]. PMID- 2967654 TI - [Study of the vascular system of the eyes in high myopia]. PMID- 2967655 TI - [Participation of the heart and blood vessels in the regulation of blood pressure after administration of angiotensin II]. PMID- 2967656 TI - [Functional tests in the selection of candidates for saturation diving]. PMID- 2967657 TI - [Elastogenesis in various conditions of culture]. PMID- 2967658 TI - Full thickness abdominal wall resection for recurrent and metastatic neoplasms. A report of three cases. AB - Full thickness infiltration of the abdominal wall by intraabdominal cancer, especially after effective prior therapy, is frequently considered a sign of inoperability. Such patients are treated with irradiation, chemotherapy, or locally destructive means (laser, cryosurgery) with limited success. Full thickness resection of portions of the abdominal wall has not been widely used in such settings because of concerns regarding abdominal wall reconstruction and also because of the perceived noncurative nature of such "radical" procedures. While long-term prognosis in patients with abdominal wall infiltration by cancer depends upon various factors, technical considerations regarding reconstruction need not prevent effective local palliative therapy. Bridging large defects can be successfully accomplished by using available synthetic material of a durable, inert nature. Polyprophylene (Marlex) mesh, a readily available and inexpensive inert fabric, meets all desirable features for such use. This paper details the successful treatment of three patients in whom recurrent carcinoma infiltrated the full thickness of the abdominal wall and caused intractable pain. In all three, full thickness portions of the abdominal wall were resected, segments of intestine and of urinary bladder were also excised, and the defects were closed using a double layer of polypropylene mesh. Complete relief of pain was accomplished and the cosmetic and functional results were excellent. Illustration A depicts procedure in patients one and two. Illustration B depicts procedure in patient three. PMID- 2967659 TI - Tremor induced by trimethoprim-sulfamethoxazole in patients with the acquired immunodeficiency syndrome (AIDS). PMID- 2967660 TI - Azathioprine and hypersensitivity vasculitis. PMID- 2967661 TI - Danazol and erythema multiforme. PMID- 2967662 TI - Hormonal intervention: "buffer hormones" or "state dependency". The role of dehydroepiandrosterone (DHEA), thyroid hormone, estrogen and hypophysectomy in aging. PMID- 2967663 TI - Psychoimmunologic and endorphin function in the aged. PMID- 2967664 TI - Miniature implantable laser Doppler probe monitoring of free tissue transfer. AB - A 2.5-mm fiber-optic laser Doppler flowmetry probe has been applied in an experimental dog model as well as in 5 clinical cases to provide continuous readout of deep tissue perfusion. The rectus abdominis muscle in the dog was used for the experimental verification of the probe, which has a linear correlation with flow rate and a rapid response (6 seconds) to arterial occlusion and venous occlusion (20 seconds). Four of the 5 free tissue transfers survived with the laser Doppler instrument correctly identifying the lack of flow, both intraoperatively and postoperatively, in the failed flap. This probe greatly extends the versatility of laser Doppler flow measurement in the clinical setting and may be nearly an ideal probe for monitoring free tissue transfer, particularly muscle. PMID- 2967665 TI - Mucopolysaccharide polysulphate cream in the prevention of pressure sores--a double blind study. AB - Early pressure areas in 38 cases (33 patients) were randomly treated twice daily with topical applications of either mucopolysaccharide polysulphate (MPS) or the cream base. The patients were assessed weekly for 4 weeks, the main parameter being change in the area of the pressure area. In the MPS group a significant reduction in size was obtained during the 2nd week of treatment, while in the control group a significant reduction was not obtained until week 4. At the end of the treatment, 59% (10/17) of the MPS group were fully healed compared with 21% (4/19) of the control group, the difference being statistically significant, p less than 0.05. MPS resembles heparin chemically, and the results could be attributed to an improvement in the microcirculation and nutrition of the perivascular tissue through an inhibition of microthrombi and perivascular fibrin deposition. PMID- 2967666 TI - Motor unit synchronization in physiologic, enhanced physiologic, and voluntary tremor in man. AB - Synchronization between pairs of single motor units simultaneously recorded from wrist extensor muscles was quantitated in 3 normal subjects during physiologic tremor (PT), beta-adrenergically enhanced physiologic tremor (EPT), and fast voluntary wrist flexion-extension movements mimicking tremor (VT). Cross correlation histograms generated from the two spike trains of each motor unit pair demonstrated central or paracentral peaks in 13/19 recordings during PT, 22/36 during EPT, and 6/7 during VT. Relative peak area was used as a quantitative index of synchronization between the two motor units of each pair. It was lowest in PT, progressively increased in EPT as tremor amplitude increased, and highest in VT. In PT and lower amplitude EPT, the synchronization indexes were higher between motor units that discharged at the same or nearly the same frequency. In contrast, in higher amplitude EPT and VT, motor units with different firing frequencies were sometimes strongly synchronized as a consequence of double discharges in faster-firing motor units that had burst repetition rates in the range of slower-firing motor units discharging as singlets. Greater motor unit synchronization with increasing tremor amplitude in EPT may be secondary to a simultaneous increase in muscle spindle afferent activity from the tremulous muscle. Greatest synchronization in VT presumably reflects near maximal supraspinal and segmental common synaptic input onto motoneurons that generate VT. These results support a longstanding hypothesis that synchronization of motor units is the physiological basis for higher amplitude tremor. PMID- 2967667 TI - Striatal calcium channel antagonist receptors in Huntington's disease and Parkinson's disease. AB - The density of calcium channel antagonist receptors labeled by (+)-[3H]PN 200-110 was reduced by 75% in striata from patients with Huntington's disease, but unchanged in patients with Parkinson's disease, compared with control subjects. These receptors are therefore likely to be localized to neurons with cell bodies in striatum, rather than nigrostriatal nerve terminals or glia, and their loss may contribute to the pathophysiology of basal ganglia disorders. PMID- 2967668 TI - Electroconformational coupling: how membrane-bound ATPase transduces energy from dynamic electric fields. PMID- 2967669 TI - Potent antipneumocystis and antitoxoplasma activities of piritrexim, a lipid soluble antifolate. AB - Piritrexim, a lipid-soluble antifolate, was evaluated for its activity against Pneumocystis carinii and Toxoplasma gondii. The concentration of piritrexim needed to inhibit 50% of the catalytic activity of P. carinii dihydrofolate reductase (DHFR) was 19.3 nM, and that for T. gondii DHFR was 17.0 nM, concentrations that were 40- to over 1,000-fold less than those needed for the inhibition of activity by trimethoprim and pyrimethamine, the antifolates conventionally used in treating these organisms. Piritrexim was able to inhibit replication of T. gondii in a mouse peritoneal macrophage model at concentrations of 0.1 to 1.0 microM. Leucovorin, a reduced folate that can bypass the inhibition of DHFR by antifols in mammalian cells but not in protozoa, did not affect the ability of piritrexim to inhibit T. gondii replication. The addition of sulfadiazine, which alone was ineffective, to piritrexim allowed inhibition of T. gondii replication at lower concentrations of piritrexim than when piritrexim was used alone. These results suggest that piritrexim, alone or combined with a sulfonamide, may be a highly potent antitoxoplasma and antipneumocystis agent that could provide major pharmacologic and clinical advantages over available agents. PMID- 2967670 TI - Synergistic activity between vancomycin or teicoplanin and gentamicin or tobramycin against pathogenic diphtheroids. AB - The in vitro activities of vancomycin and teicoplanin alone and in combination with gentamicin or tobramycin were studied by time-kill techniques with 11 strains of pathogenic diphtheroids (Corynebacterium group JK). The activities of vancomycin and teicoplanin were similar (MIC for 90% of strains tested [MIC90], 1 microgram/ml), as were those of gentamicin and tobramycin (the MIC90 was 1 microgram/ml for five aminoglycoside-susceptible strains, and the MIC90 was greater than 1,024 micrograms/ml for six aminoglycoside-resistant strains). No consistent synergistic killing could be demonstrated by the combination of glycopeptide and aminoglycoside antibiotics at arbitrarily chosen concentrations within the range of clinically achievable levels. However, by careful adjustment of both vancomycin and gentamicin concentrations within a narrow range below the MIC of each antibiotic, synergistic killing could be seen with an aminoglycoside susceptible strain but not with an aminoglycoside-resistant strain. Synergism between glycopeptide and aminoglycoside antibiotics occurs with some diphtheroid organisms, but it may not be clinically relevant. PMID- 2967671 TI - Activity of trospectomycin against Bacteroides fragilis and other Bacteroides species. AB - The in vitro activity of trospectomycin (U-63366; 6'-n-propyl spectinomycin pentahydrate sulfate) was evaluated against 189 clinical isolates of the Bacteroides fragilis group and 65 Bacteroides species isolates. At less than or equal to 8 micrograms/ml, the activity of trospectomycin compared favorably with those of clindamycin and cefoxitin against B. fragilis, Bacteroides distasonis, and Bacteroides vulgatus, and there was no cross resistance to these three drugs among the strains of the B. fragilis group. All the Bacteroides species were susceptible to trospectomycin. The results of this in vitro study indicate that trospectomycin possesses excellent activity against Bacteroides species. PMID- 2967672 TI - Comparison of virulence factors and R plasmids of Salmonella spp. isolated from healthy and ill swine. AB - The antibiotic resistance and virulence profiles of Salmonella spp. isolated from healthy (group 1) and ill (group 2) swine were compared. Parameters studied included colicin and siderophore production; mannose-sensitive hemagglutination of erythrocytes; resistance to the lethal effect of serum complement; resistance to antibiotics; and the transmissibility of these characteristics to recipient organisms. Group 1 (19 isolates) had 14 serotypes, and group 2 (20 isolates) had 2 serotypes. Isolates from group 2 were resistant to more antibiotics and had a greater ability to hemagglutinate erythrocytes and transfer R plasmids to recipient organisms, but a lesser ability to produce siderophore than group 1. All 39 isolates resisted the lethal effects of serum complement. Colicin was produced by 1 of 19 from group 1 and 0 of 20 from group 2. A donor Escherichia coli isolated from a pig with enteritis transferred R plasmids to 62% of group 1 and 0% of group 2 Salmonella spp. when they were used as recipient organisms. A transconjugant from the mating of donor E. coli to a group 1 Salmonella spp. was further able to pass an R plasmid to recipient E. coli and salmonellae. Plasmid isolation from group 1 yielded 1 of 19 strains with a 56-megadalton plasmid, while 20 of 20 strains from group 2 contained three to five plasmids from 2.4 to 60 megadaltons in size. PMID- 2967673 TI - Characterization of rat heart plasma membrane Ca2+/Mg2+ ATPase. AB - The Ca2+/Mg2+ ATPase of rat heart plasma membrane was activated by millimolar concentrations of Ca2+ or Mg2+; other divalent cations also activated the enzyme but to a lesser extent. Sodium azide at high concentrations inhibited the enzyme by about 20%; oligomycin at high concentrations also inhibited the enzyme slightly. Trifluoperazine at high concentrations was found inhibitory whereas trypsin treatment had no significant influence on the enzyme. The rate of ATP hydrolysis by the Ca2+/Mg2+ ATPase decayed exponentially; the first-order rate constants were 0.14-0.18 min-1 for Ca2+ ATPase activity and 0.15-0.30 min-1 for Mg2+ ATPase at 37 degrees C. The inactivation of the enzyme depended upon the presence of ATP or other high energy nucleotides but was not due to the accumulation of products of ATP hydrolysis. Furthermore, the inactivation of the enzyme was independent of temperature below 37 degrees C. Con A when added into the incubation medium before ATP blocked the ATP-dependent inactivation; this effect was prevented by alpha-methylmannoside. In the presence of low concentrations of detergent, the rate of ATP hydrolysis was reduced while the ATP dependent inactivation was accelerated markedly. Both Con A and glutaraldehyde decreased the susceptibility of Ca2+/Mg2+ ATPase to the detergent. These results suggest that the Ca2+/Mg2+ ATPase is an intrinsic membrane protein which may be regulated by ATP. PMID- 2967674 TI - Efficiency of acitretin in the treatment of cutaneous lupus erythematosus. AB - Acitretin (etretin [Ro-10-1670]) is the major metabolite of etretinate with a much shorter elimination half-life. The drug was used in the treatment of 20 patients who had cutaneous lupus erythematosus. All patients responded to treatment, but in five, the result was unsatisfactory. In 15 patients, an excellent (total clearing) or good response (marked reduction of all lesions) was seen. In seven of them, acitretin was superior to previous therapy with antimalarials and/or systemic corticosteroids. In particular, five of six patients with subacute cutaneous lupus erythematosus showed complete clearing of their lesions, usually within two to four weeks. Side effects were the same as with etretinate. It is concluded that acitretin is a highly effective and well tolerated drug in the treatment of cutaneous lupus erythematosus. PMID- 2967675 TI - Contact allergy to clobetasol propionate. AB - Two cases of contact allergy to clobetasol propionate are described. Neither patient had positive epicutaneous tests to other structurally similar fluorinated steroids or to other steroids with the propionate ester, but both had multiple positive tests within the European standard battery. Comparison with previously reported cases reveals that multiple contact allergies are a frequent finding in patients with allergy to this topical steroid. PMID- 2967676 TI - Allergic contact dermatitis to Alstroemeria. AB - Two female florists developed dermatitis of the fingertips. Patch testing revealed allergic contact dermatitis to the flower, Alstroemeria, used in floral arrangements. They had positive patch tests to portions of Alstroemeria, and to tuliposide A, the allergen in this plant. Vinyl gloves were not helpful since tuliposide A readily penetrates through these gloves. Nitrile gloves may be protective since they prevented positive patch test to tuliposide A. PMID- 2967677 TI - Skin lesions due to exposure to methyl bromide. AB - Six patients were occupationally exposed to high concentrations of methyl bromide during a fumigation procedure using adequate airway protection. Within a few hours all patients developed skin lesions, consisting of sharply demarcated erythema with multiple vesicles and large bullae. There was a striking predisposition for parts of the skin that were relatively moist or subject to mechanical pressure, such as axillae, groin, and abdomen. Microscopically, early skin lesions revealed necrosis of keratinocytes, severe edema of the upper dermis, subepidermal blistering, and diffuse infiltration of neutrophils and, to a lesser degree, eosinophils. Two patients developed an urticarial rash approximately one week after the exposure. On histologic examination, these late lesions showed combined features of a spongiotic dermatitis and urticaria. No immunopathologic manifestations were observed. In all patients, the skin returned to normal after four weeks, except for some residual hyperpigmentation. Plasma bromide levels after exposure strongly suggested percutaneous absorption of methyl bromide. PMID- 2967679 TI - Leu-11b (CD16) antigen expression on human keratinocytes. PMID- 2967680 TI - Surgical removal of guidewire fragment following transluminal coronary angioplasty. AB - Following percutaneous transluminal coronary angioplasty, wire fragments retained within the patient usually can be removed without an operation. We report a case requiring surgical removal of a guidewire fragment from the distal circumflex coronary artery. PMID- 2967678 TI - Effect of an orally administered arotinoid, Ro 15-0778, on sebum production in man. AB - 1,2,3,4-Tetrahydro-1,1,4,4-tetramethyl-y-[(E)-alpha-methylstyryl]- naphthalene (Ro 15-0778), a potent inhibitor of sebaceous gland activity in several animal models has been administered to male volunteers in dosages of 1, 2, 3, and 6 mg/kg/day and 2 g/day for 8 weeks. Sebum production was measured before treatment and after 4 and 8 weeks of therapy. At the dosage levels of 1, 2, 3, and 6 mg/kg/day there was no decrease in sebum secretion. With the dosage of 2 g/day, a significant decrease in sebum secretion was seen after 8 weeks of treatment with Ro 15-0778. PMID- 2967681 TI - Measures of central tendency, variability, and relative standing in nonnormal distributions: alternatives to the mean and standard score. AB - There are two major disadvantages to reporting test results in standardized scores such as z-scores or t-scores when describing individuals with disabilities. (1) Raw scores on many tasks from disabled individuals are notoriously nonnormal. They are asymmetric, in that they are skewed toward poor scores. (2) Low z-scores give a false impression of dysfunction because able bodied subjects are often grossly overqualified for the application in question; they may have levels of strength or quickness several orders of magnitude greater than what is needed to do the assessment task. A recommended alternative is to describe the performance of disabled individuals using nonparametric statistics, and to report scores in boxplots, showing the extremes, the median, and the quartiles. Such points are easy to calculate and to interpret, and they are robust against outliers. PMID- 2967682 TI - Self-management for medication reduction in chronic low back pain. AB - It has been demonstrated that pain relief is seldom produced by medication or surgical methods where there is evidence of emotional disturbance, as indicated by the MMPI. A program that attempts to engender a high level of patient responsibility in a population of chronic low back pain patients is described. Self-managed reduction of drug dependence is a major component of this program. The data indicate that the program produces a significant reduction in dependence on opiates, derivatives, synthetic opiates, hypnotics, sedatives, tranquilizers, and analgesics. Follow-up data (with attrition controlled) at six months and 12 months postdischarge do not provide any evidence for deterioration (ie, return to pretreatment levels of drug dependence). Thus, it appears that the programmatic impact is stable over at least a 12-month period postdischarge. Implications of these findings for the low back pain population, as well as other chronic pain populations, are discussed. PMID- 2967683 TI - Lack of inhibition of glycolytic enzymes by the neurotoxic organophosphorus compounds mipafox and methamidofos. AB - Preincubation with Mipafox and Methamidofos as well as Paraoxon (used as control) did not cause inhibition of hexokinase, phosphofructokinase, glyceraldehyde-3 phosphate dehydrogenase and lactate dehydrogenase. This is in contrast with the inhibition of glycolysis by other neurotoxic compounds (hexacarbons, acrylamide, carbon disulfide). PMID- 2967684 TI - [Analysis of the activities of pathology departments in Obninsk during the past 20 years (1966-1985)]. AB - This review of data from pathology departments in the town of Obninsk has shown that over the last 20 years the most frequent causes of death, among males and females alike, were cardiovascular diseases (44%), with malignant neoplasms ranking second (30%) and causing almost twice as many deaths among males as among females. From 63% to 85% of deaths due to malignancy were from epithelial tumors, particularly of the stomach and lungs. A steady increase in the volume of autopsy material and especially of material from intravital morphologic examinations was apparent, with a tendency to a rise in the proportion of biopsy specimens relative to operative specimens, although the latter still predominate. PMID- 2967685 TI - [Current objectives in the organization and progress in pathological services in rural public health]. AB - Specific conditions of pathological service in rural areas confined to a single prosectorium per two or more distant hospitals with limited facilities for dissection and biopsy processing when one specialist on the staff, call for an appropriate organizational frame-work. It involves regional centralization, provision with travelling laboratories for autopsy and rapid biopsy evaluation, additional personnel admission to reduce overwork. A standard notice is developed for registration of the discrepancy between a clinical and pathological diagnosis aimed at perfection of statistically significant estimation of misdiagnosis with a lethal outcome. To ensure a radical change in rural pathologic service and its material supply, it is necessary to establish its independent status under the control of central regional or republican pathologic bureaus and relevant all union research institute. PMID- 2967686 TI - [Vladimir Georgievich Garshin (1887-1956)]. PMID- 2967687 TI - Effect of medroxyprogesterone acetate on peritoneal adhesion. PMID- 2967688 TI - Laparoscopy as a diagnostic and therapeutic technique in uterine perforations during first trimester abortions. PMID- 2967689 TI - Measurement of placental blood flow by the thermocouple method in comparison with the electromagnetic flowmeter method. PMID- 2967690 TI - Evaluation of hearing handicap: a critique of Ward's position. PMID- 2967691 TI - Effect of propranolol upon protein and proteolytic synthesis activity in hypertrophic myocardium. AB - The aim of the study was to investigate the effect of propranolol upon protein synthesis and degradation processes in cell-free subfractions of rat myocardium in experimental cardiac hypertrophy induced by aortic stenosis. It was found that hypertrophy stimulates incorporation of 3H-amino acids by the postmitochondrial supernatant (PMS) by 17 +/- 4% (mean +/- SE). Propranolol 10 6M inhibited protein synthesis in the control and experimental groups by 37 +/- 5% and 34 +/- 7%, respectively. At a concentration of 5 X 10 6M, propranolol-induced inhibition in each group was 74 +/- 4% and 58 +/- 5%, respectively. The activity of neutral proteases in PMS of hypertrophic hearts increased up to 255 +/- 29 IU in comparison to 154 +/- 10 IU in controls; in the cytosol it was unchanged. Propranolol had no effect upon the activity of proteases in both fractions. Activity of calcium-activated neutral proteases (CANP) remained at the same level in control and hypertrophic hearts and was unaffected by propranolol. PMID- 2967693 TI - [Effect of segmental mobilization on lumbar vertebrogenic pain syndromes]. PMID- 2967692 TI - Age-dependent changes of relaxation and its load sensitivity in rat cardiac muscle. AB - The relaxation phase and its load dependence were studied in papillary muscles isolated from the left ventricle of rats of the following ages: 20 days, 2, 8, 18, and 24 months. The myofibrillar ATPase activity and the force-velocity relation were determined in each age group in order to characterize the kinetic properties of the contractile material. Both shortening velocity and myofibrillar ATPase activity showed a progressive reduction with maturation and aging. This observation suggested an age-dependent decrease in cross bridge formation rate. The relaxation phase was characterized by its duration and the maximum rate of tension decline in isometric conditions, and by the speed of relengthening in isotonic conditions. Relaxation became faster and of shorter duration with maturation from 20 days to 2 months and then became slower and of longer duration with further maturation and aging. The sensitivity of relaxation to changes in length or load was evaluated by measuring how much earlier tension declined in the presence of a given length change. An increase in load sensitivity of relaxation was observed during maturation from 20 days to 8 months. This increase was followed by a reduction during aging from 8 to 24 months. Such a biphasic trend of the age-related changes in load sensitivity of relaxation could result from the interplay between the progressive decrease in cross bridge formation rate and a reduction in activation decay rate. The latter was suggested by the prolongation of the relaxation phase and by the maintenance of developed tension during aging. PMID- 2967695 TI - Pharmacological profile of 8-amino octanoic acid substituted atrial natriuretic factor analogs. AB - A series of six analogs of rat atrial natriuretic factor have been prepared by the solid-phase method. The modified analogs contain 8-amino octanoic acid (a simple alkyl spacer) in place of selected tripeptides. Binding affinity to cultured aortic smooth muscle cell membranes suggests that the sequence Arg11 Gly16 is important for binding. Vasorelaxant activity on serotonin contracted rabbit aortic rings indicates that the Phe8-Gly16 sequence must be present for vasorelaxation. In anesthetized rats, the natriuretic and diuretic effects of an IV bolus dose correlate with in vitro vasodilatory activity. The alkyl spacer approach provides a facile method to quickly determine key regions of a large peptide involved in molecular recognition. PMID- 2967694 TI - Xanthine dehydrogenase expression in Neurospora crassa does not require a functional nit-2 regulatory gene. AB - Xanthine dehydrogenase (XDH) is the initial enzyme in the purine catabolic pathway of N. crassa. Secondary nitrogen sources such as purines are metabolized when preferred sources of reduced nitrogen (ammonium or glutamine) are unavailable. XDH synthesis is regulated by glutamine repression and uric acid induction. The nit-2 locus is believed to encode a trans-acting positive regulator essential for the expression of genes encoding enzymes involved in secondary pathways of nitrogen acquisition, such as XDH and nitrate reductase. However, immunoblot analyses and enzyme assays reveal that XDH protein is synthesized and XDH activity is expressed in nit-2 mutants. Nevertheless, XDH responds to nitrogen metabolite repression. The generality that nit-2 is an obligate control element in nitrogen metabolite repression is questioned. Additionally, mutants defective in XDH activity, namely, xdh-1 and the molybdenum cofactor mutants nit-1, -7, -8 and -9, are observed to grow on xanthine but not hypoxanthine. PMID- 2967696 TI - Guanine nucleotide-, and inositol triphosphate-induced inhibition of the CA2+ pump in rat heart sarcolemmal vesicles. AB - The Ca2+ pump of rat heart sarcolemma has been studied via its ATP-dependent Ca2+ transport and (Ca2+ + Mg2+)-dependent ATPase activities. Direct incubation of the sarcolemmal vesicles with micromolar concentration of guanosine 5'-O (thiotriphosphate) (GTP gamma S) results in the reduction of Ca2+ uptake by 34 +/ 10% and ATP hydrolysis by 55 +/- 7%. Similar inhibition of the sarcolemmal Ca2+ pump is also observed with micromolar concentration of inositol trisphosphate (IP3), while GDP or inositol tetrakisphosphate (IP4) has no effect. Based on the evidence that these sarcolemmal vesicles are capable of generating IP3 upon stimulation by GTP gamma S, and that no additive effect is observed when both agents are incubated together with the membranes, it is concluded that the effect of GTP gamma S on the Ca2+ pump is mediated by IP3. The results here show for the first time that plasma membrane Ca2+ pump has a role in the primary Ca2+ signaling. PMID- 2967697 TI - High molecular weight forms of insulin-like growth factor II and its binding protein identified by protein immunoblotting. AB - Insulin-like growth factor (IGF)II is a mitogenic polypeptide which circulates in association with a binding protein(s). Immunoblotting studies were performed in human serum and indicate that:(1)a approximately 200 kDa covalently-linked IGF II/binding protein complex is antigenically related to the 30 kDa binding protein, (2)IGF-II prohormone is associated with this complex, and (3)a major portion of the IGF-II prohormone immunoreactivity in human serum is present in fractions which would not be detected by standard radioimmunoassay methods. Our data provide insight regarding the inter-relationships of IGF-II and its binding protein, and direct evidence for the presence of IGF-II prohormone in human serum. PMID- 2967698 TI - Comparison of binding affinities and calcium current inhibitory effects of a 1,4 dihydropyridine derivative (PN 200-110) in vascular smooth muscle. AB - The binding of (+) (3H) PN 200-110 to high and low affinity sites in mammalian portal vein smooth muscle membranes was characterized. Binding affinities were 0.09 and 30 nM for the high and low affinity sites, respectively, and binding site densities were 45 and 400 fmoles/mg of protein for the respective sites. (+) PN 200-110 blocked both fast and slow calcium currents in isolated cells from portal vein smooth muscle. The blockade of slow calcium current was voltage dependent as PN 200-110 bound with higher affinity to inactivated slow calcium channels (IC50 = 0.03 nM) than to resting channels (IC50 = 0.15 nM). The blockade of fast calcium current was voltage-independent (IC50 = 45 nM). The IC50 values found from electrophysiological experiments for the binding to inactivated slow and fast calcium channels are similar to the Kd values determined by radioligand binding. PMID- 2967699 TI - Pattern of repeating aromatic residues in synexin. Similarity to the cytoplasmic domain of synaptophysin. AB - Synexin was isolated from bovine liver by high resolution cation exchange chromatography and fragmented with cyanogen bromide or trypsin. Peptides were isolated and their amino acid sequences partially determined. Twenty percent of the synexin sequence was determined in one contiguous sequence of 61 residues and a nonoverlapping sequence of 20 residues. The sequence is characterized by a hexapeptide repeat of the form YPXXXX occurring eight times in series, with phenylalanine substituting for tyrosine in two positions. The intervening amino acids (X) are predominantly proline, glycine and alanine. This pattern of periodic aromatic residues suggests the presence of a novel secondary structure and is similar to repeats present in synaptophysin, gliadin and type II keratin. PMID- 2967700 TI - SnRNP core protein enrichment in the nuclear matrix. AB - The D protein (16 kDa) is part of a protein core, common to U1, U2, U5, U4/U6 small nuclear RNA containing ribonucleoprotein particles. Monoclonal antibodies reactive with the D protein were used in quantitative dot blotting and Western blotting to demonstrate that this protein was a component of salt resistant nuclear structures and was enriched greater than 3 to 5-fold in RNAase-protected nuclear matrix preparations. PMID- 2967701 TI - Identification of a protein kinase activity in rabbit reticulocytes that phosphorylates the mRNA cap binding protein. AB - The 25 kDa mRNA cap binding protein can be purified in a partially phosphorylated state and the extent of its phosphorylation appears to be regulated during heat shock and mitosis in mammalian cells. We demonstrated that a nonabundant serine protein kinase activity exists in rabbit reticulocytes that phosphorylates the 25 kDa cap binding protein in both the free (eIF-4E) and complexed (eIF-4F) state. This kinase was not inhibited by the cAMP-dependent protein kinase inhibitory peptide IAAGRTGRRNAIHDILVAA, did not phosphorylate S6 ribosomal protein, did not phosphorylate p220 of eIF-4F as protein kinase C does and no other substrates for this kinase were apparent in reticulocyte ribosomal salt wash. The molecular identity of this kinase, the specific site(s) of eIF-4E that it phosphorylates and its in vivo regulatory role remain to be studied. PMID- 2967702 TI - Interaction of permanently uncharged dopamine analogs with the D-2 dopaminergic receptor. AB - The purpose of this study was to determine if structural analogs of dopamine in which the side chain nitrogen has been replaced by a permanently uncharged monomethylsulfide, monomethylselenide or sulfoxide group are capable of binding to the striatal D-2 dopamine receptor and acting as agonists at this receptor. All the permanently uncharged dopamine analogs were found to bind to the D-2 dopamine receptor as evidenced by their abilities to inhibit significantly [3H]spiperone binding to striatal homogenates. However, the inhibition of [3H]spiperone binding by the uncharged dopamine analogs was incomplete and was almost abolished by the addition of NaCl (125 mM) to the incubation medium or by the addition of dopamine or quinpirole at a concentration that that saturates the high-affinity state of the D-2 dopamine receptor. These effects of NaCl, dopamine and quinpirole suggest that the uncharged dopamine analogs bind primarily to the high-affinity state of the D-2 dopamine receptor. Whether the uncharged monomethylsulfide and sulfoxide analogs could function as dopamine agonists at the striatal D-2 dopamine receptor was assessed by determining the abilities of these compounds to inhibit the K+-evoked release of [3H]acetylcholine from striatal slices. Both the monomethylsulfide and sulfoxide analogs inhibited the K+-evoked release of [3H]acetylcholine, but this inhibitory effect does not appear to be due to the activation of the D-2 dopamine receptor since it was not reversed by the selective D-2 dopamine antagonist, sulpiride. Additionally, the uncharged monomethylsulfide and sulfoxide dopamine analogs were found to antagonize the ability of apomorphine to inhibit the K+-evoked release of [3H]acetylcholine, but this antagonistic effect does not appear to be due to the reversible blockade of the D-2 dopamine receptor since it was not reduced by increasing the concentration of apomorphine. Therefore, while the permanently uncharged analogs of dopamine appear to bind to the high-affinity state of the D 2 dopamine receptor, they are not dopamine agonists or antagonists at the striatal D-2 dopamine receptor involved in regulating the release of acetylcholine. These results suggest that a positive charge may be a requirement for the activation of the striatal D-2 dopamine receptor. PMID- 2967703 TI - Alkylation of the liver plasma membrane and inhibition of the Ca2+ ATPase by acetaminophen. AB - Acetaminophen is activated metabolically to yield reactive species that bind covalently to liver cell macromolecules. The extent of covalent binding correlates with the occurrence and severity of hepatic necrosis. We reported previously [J. O. Tsokos-Kuhn, E. L. Todd, J. B. McMillin-Wood and J. R. Mitchell, Molec. Pharmac. 28, 56 (1985)] that active Ca2+ accumulation of isolated liver plasma membranes is decreased 60-75% after a hepatotoxic dose of acetaminophen in vivo. We now report that the protein of isolated liver plasma membranes was substantially labeled with drug metabolites after administration of [3H]acetaminophen. There was no increase in passive membrane permeability that might cause diminished Ca2+ accumulation. Intravesicular volume and relative purity of the vesicle preparations after acetaminophen were not different from controls. However, (Ca2+,Mg2+)-ATPase, a possible biochemical expression of the Ca2+ pump, was decreased 31% (P less than 0.025) after acetaminophen treatment. ATPase activity in both control and treated groups was enhanced by isolating membranes in the presence of 5 mM reduced glutathione (GSH), but the effects of drug treatment were not reversed. A similar effect of GSH on Ca2+ accumulation was observed previously [J. O. Tsokos-Kuhn, E. L. Todd, J. B. McMillin-Wood and J. R. Mitchell, Molec. Pharmac. 28, 56 (1985)]. These data are consistent with a hypothesis wherein alkylation of membrane proteins by reactive acetaminophen metabolites is a factor in the onset of hepatic necrosis after acetaminophen. They are not consistent with an oxidative stress hypothesis where thiol S thiolation of membrane components is postulated to produce altered membrane permeability or thiol-reversible alterations in membrane protein structure and enzymatic function. PMID- 2967704 TI - Lack of effect of insulin in hepatocytes isolated from streptozotocin-diabetic male rats. AB - Diabetes mellitus is known to affect drug and steroid metabolism in the rat liver. Recently we have demonstrated that in-vitro insulin addition to hepatocytes obtained from normal male rats showed a significant dose-related increase in androstenedione metabolism. We have extended our study this time by using 3- and 21-days streptozotocin (STZ) diabetic and insulin-treated STZ diabetic male rats. Hepatocytes from 3- and 21-days STZ-diabetic rats were resistant to the effect of insulin while insulin-treated diabetic rats indicated partial restoration of insulin effect. Since insulin resistance is a characteristic feature of type II diabetes mellitus, we would like to suggest that STZ-diabetic rats may be a model for type II diabetes mellitus. PMID- 2967705 TI - The phage Mu 'late' gene transcription activator, C, is a site-specific DNA binding protein. PMID- 2967706 TI - Serum keratan sulfate levels in osteoarthritis patients. AB - Serum levels of keratan sulfate (KS), measured by an enzyme-linked immunosorbent inhibition assay, were found to be significantly higher in 31 patients with hypertrophic osteoarthritis (OA) than those in 41 adults without joint disease. Seventy-seven percent of patients with OA, but only 12% of control subjects, had serum levels which were more than 1 SD above the mean of the control group. Following replacement of a single osteoarthritic hip joint, serum KS levels decreased, at first, in all patients. Subsequently, the concentration of serum KS progressively increased; 6 months following surgery, KS levels were similar or close to the preoperative levels in virtually all patients. The results suggest that patients with hypertrophic OA may have a generalized imbalance of cartilage proteoglycan metabolism. Measurements of serum KS are likely to prove most useful in studying this particular subset of patients with generalized OA. PMID- 2967707 TI - Epidemiology of chronic musculoskeletal disorders. PMID- 2967708 TI - Hyperplasia of the mesenterial windows precedes that of the small gut in the streptozotocin-diabetic rat. AB - Hyperplastic growth of the mesenterial windows abutting the small gut occurs in lactating rats (Bergstrom and Norrby 1988) and chronically diabetic rats (Norrby et al. 1983). In the present study, early events in the mesenterial windows and the small gut in streptozotocin-diabetic rats were examined. The area of the mesenterial windows had already increased significantly on day 1 and hyperplasia in terms of increased DNA content, as well as an increase in histamine content (a mast-cell marker), was established from day 2 of diabetes. The increase in total mesenterial window content of DNA, histamine and protein was roughly linear and parallel from day 2 to day 19. The small-gut circumference increased transiently on day 1, but the small-gut mucosal volume was unaffected on days 1 and 2. The small-gut wet weight increased significantly from day 5, whereas elongation was not observed until day 19. The difference in time between the appearance of hyperplasia and the growth of the mesenterial windows and their adjoining gut and the rate with which the hyperplasia proceeds in the two tissues indicate that the regulatory mechanisms of early hyperplastic growth in these tissues are not identical. The factor(s) causing mesenterial window growth and hyperplasia is/are as yet unknown. PMID- 2967709 TI - Application of dynamic computed tomography to physiologic imaging of renal artery stenosis before and after angioplasty. AB - This study describes the application of dynamic Computed Tomography (CT) of the kidneys to the diagnosis and evaluation of the physiology of renal artery stenosis before and after angioplasty. The CT imaging parameters are discussed in relationship to findings from other imaging modalities, which include renal arteriography, intravenous digital subtraction angiography, and radionuclide renography. Time density graphs of the renal cortex and medulla were obtained by the use of region of interest cursors, and the corticomedullary junction time (CMJT) was determined. The CMJT was prolonged in the setting of renal artery stenosis and was improved after angioplasty. Simple visual inspection alone provided sufficient information to diagnose renal artery stenosis. The pathophysiology of renal artery stenosis and the meaning of the abnormal findings is discussed. Dynamic CT provides a unique noninvasive means for cross-sectional physiologic imaging. The role of dynamic CT in the evaluation of renovascular hypertension and physiologic imaging in other areas requires further investigation. PMID- 2967710 TI - The dental management of patients with ankyloblepharon (AEC) syndrome. PMID- 2967711 TI - Comparison of sufentanil-oxygen and fentanyl-oxygen anaesthesia for coronary artery bypass grafting. AB - Haemodynamic variables were compared in 40 adults undergoing coronary artery bypass grafting during anaesthesia induced with either sufentanil 5 micrograms kg 1 or fentanyl 25 micrograms kg-1 in combination with pancuronium 0.1 mg kg-1. Further doses of sufentanil 2.5 micrograms kg-1 or fentanyl 12.5 micrograms kg-1 were given before skin incision and again before sternotomy. All patients were receiving beta-adrenoceptor blocking therapy. Satisfactory induction of anaesthesia was produced with both drugs and opioid supplementation prevented any marked haemodynamic response to skin incision and to sternotomy. Following induction of anaesthesia, sufentanil produced the greater decrease in mean arterial pressure and left ventricular stroke work index which continued throughout the study. This suggests that, in the doses used in this study, sufentanil is preferable to fentanyl in patients with coronary artery disease. PMID- 2967712 TI - Nitrous oxide and day-case laparoscopy: effects on nausea, vomiting and return to normal activity. AB - Patients admitted for day-case laparoscopy were assigned randomly to receive nitrous oxide-oxygen or oxygen, with enflurane, during a standard anaesthetic technique. Postoperative morbidity, in particular nausea and vomiting, and ability to resume normal activity were assessed over the ensuing 48 h. Supplementary administration of propofol during the operative procedure was required significantly more often (P less than 0.05) in the absence of nitrous oxide. There was no significant difference in the incidence of vomiting before discharge when nitrous oxide was omitted. The incidence and severity of nausea over the 48 h following operation was similar in both groups. There was no difference in analgesic or anti-emetic requirements before discharge and the time taken to resume normal activity was similar. It is concluded that nitrous oxide may be avoided readily in day-case laparoscopy without affecting postoperative morbidity or time taken to return to "street fitness" and normal activity. PMID- 2967713 TI - High frequency jet ventilation. PMID- 2967715 TI - Influence of age and severity of dermatitis on the percutaneous absorption of hydrocortisone in children. AB - The results of 55 4-h hydrocortisone absorption tests in 38 children with atopic or seborrhoeic dermatitis were analysed to evaluate the effect of age and severity of the dermatitis on percutaneous absorption of hydrocortisone. The children were divided into three groups on the basis of the severity of the dermatitis. The absorption of hydrocortisone caused a significantly higher mean rise of serum cortisol in 20 children with severe dermatitis (Group A) than in 17 children with moderate dermatitis (Group B). The mean post-application rise of serum cortisol in 18 children with mild dermatitis (Group C) was significantly lower than in the children with moderate dermatitis. There was a significant negative linear correlation between age and the post-application rise of serum cortisol in Groups A and C. In these groups the mean post-application rise of serum cortisol was significantly higher in children aged under 18 months than in children aged 18 months or over. Severe widespread dermatitis and an age under 18 months are two relevant risk factors in the topical use of hydrocortisone. PMID- 2967714 TI - Prognostic value of Langerhans cells in the epidermis of HIV patients. AB - We measured the number of Langerhans cells in clinically normal epidermis and the number of T4 lymphocytes in the plasma in 24 patients infected with HIV, and related these to the stage of the disease. A correlation is demonstrated between the numbers of Langerhans cells and T4 lymphocytes. Numbers of Langerhans cells were lower in patients with disease stages III and IV than in those with disease stage II. Thus Langerhans cell numbers could be of prognostic value in this disease. PMID- 2967716 TI - Drug-induced pseudolymphomatous skin reactions. AB - We present two patients with generalized and three with localized skin eruptions due to various non-anticonvulsant drugs, with a histological picture of a pseudolymphoma of the skin. Cessation of the causative drugs resulted in disappearance of the lesions in all cases. PMID- 2967717 TI - Changes in the plasma levels of vitamin K-dependent proteins C and S and of C4b binding protein during pregnancy and oral contraception. AB - The plasma concentrations of protein S, protein C and C4b-binding protein (C4BP) were analysed during pregnancy, in the postpartum period and in women using oral contraceptives. Free protein S, measured after precipitation of the C4BP-protein S complexes with 5% PEG 6000, was found to be 8.3 mg/l in the control group, which represents 36.3% of the total plasma protein S content (average 23.5 mg/l). The concentration of protein S was significantly decreased during pregnancy, the lowest levels occurring in the second trimester (14.8 mg/l). The values returned to normal within a few days after delivery. The concentration of free protein S was also decreased, down to an average of 3.7 mg/l at delivery, and did not return to normal within the first week postpartum. The mean concentration of protein S in women using oral contraceptives decreased to 17.7 mg/l and the free fraction went down to 6.6 mg/l. Unlike that of protein S, the plasma concentration of protein C increased during pregnancy, reaching a maximum of 135% in the second trimester. Also, it was significantly higher in the postpartum period and in women using oral contraceptives, than in controls. The level of C4BP was increased throughout pregnancy, with a maximum of 143.4% at delivery. These changes in the plasma levels of proteins C and S during pregnancy indicate that the two proteins differ in the regulation of their synthesis. The major decrease in the level of free protein S may predispose to thrombotic episodes during pregnancy, whereas the increased level of protein C may have the reverse effect. These results indicate the importance of taking into account the normal changes in the plasma levels of protein C and S during pregnancy and the use of oral contraceptives, when evaluating patients with increased risk of thromboembolic disease. PMID- 2967718 TI - Plasma concentrations of C4b-binding protein and vitamin K-dependent protein S in term and preterm infants: low levels of protein S-C4b-binding protein complexes. AB - We have determined the plasma concentrations of protein S and C4BP in 25 term and 26 preterm infants by radioimmunoassay. Both the total concentration and the concentration of free protein S were quantified. The concentration of C4BP was very low in preterm infants (mean 6% of the adult level). In term infants, the level had increased to a mean of 18%. Total protein S was decreased both in preterm and term infants, 4.0 mg/l and 6.8 mg/l respectively, as compared to the mean adult concentration, 20.6 mg/l. In preterm infant plasma, free protein S was the predominant (85%) form, probably due to the very low C4BP level. In plasma from term infants, free protein S represented 68% of the total protein S, the corresponding value in adult controls being 37%. The plasma concentration of free protein S in preterm and term infants was 3.3 mg/l and 4.6 mg/l, respectively (mean adult value 7.6 mg/l). These results demonstrate that, while the total protein S concentration in preterm and term infants was very low in comparison to the adult level, the difference in the concentration of the anticoagulant, active, free form of protein S between infants and adults was less pronounced. PMID- 2967719 TI - Structural characterization of heparan sulfate proteoglycan subclasses isolated from bovine aortic endothelial cell cultures. AB - Labeled heparan sulfate proteoglycans (HSPG) were isolated from wounded and confluent cultures of bovine aortic endothelial cells by nondegradative extraction with 4 M guanidine hydrochloride and detergent. HSPG were separated from more highly charged chondroitin or dermatan sulfate proteoglycans by ion exchange chromatography, and subclasses of different hydrodynamic size were isolated by gel filtration. Three major subclasses of HSPG were characterized structurally with respect to the presence and relative size of protein core, the presence and amount of nonsulfated oligosaccharide, and size and structure of heparan sulfate (HS) chains. The largest (600-800-kDa) HSPG subclass (I), isolated from cell layers and media of confluent cultures, bears 38-kDa HS chains on an apparently heterogeneous class of relatively large glycoprotein cores. HSPG II (150-200 kDa), isolated from cell layer or media, has 22-kDa HS chains and smaller core glycoproteins (less than 50 kDa). HSPG III, the subclass of smallest hydrodynamic size, has 13-kDa HS chains and a glycopeptide core of less than 15 kDa. All subclasses bear varying proportions of non-sulfated oligosaccharides of similar sizes. Comparisons of HS chain structure indicated that the different subclasses have similar proportions (49-55%) of N-sulfate, with both O-sulfate and highly N-sulfated blocks of disaccharide distributed similarly along HS chains. In addition, HS chains from subclasses II and III contain sequences that are insensitive to periodate oxidation or heparitinase digestion, suggesting that they contain increased proportions of iduronate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967720 TI - Lymphocyte subsets of left-handers. PMID- 2967721 TI - Behaviorally conditioned enhancement of delayed-type hypersensitivity in the mouse. AB - Cyclophosphamide (CY), previously used to condition suppression of a number of different immune responses, was used to condition an enhancement of a delayed type hypersensitivity (DTH) response in mice. Three weeks before sensitization with sheep red blood cells (SRBC), mice were conditioned by pairing their consumption of a saccharin solution (SAC) with injection of CY. Two, three, and four days after sensitization (Day 0), animals were reexposed to SAC and, on Day +3, they were injected with a low dose of CY. This dose suppressed the DTH response to a challenge with SRBC on Day +4 in both conditioned and control animals. Following rechallenge with SRBC on Days +11 and +18, however, DTH responses were enhanced. Conditioned mice that had been reexposed to SAC showed a significantly greater enhancement than either nonconditioned animals or conditioned mice that were not reexposed to SAC. Thus, enhancement as well as suppression of immune responses can be conditioned with CY. PMID- 2967722 TI - In vivo effects of beta-endorphin on lymphocyte proliferation and interleukin 2 production. AB - Experiments were undertaken in rats to investigate the effects of in vivo infusion of beta-endorphin (BEP) on subsequent Con A-induced proliferation and interleukin 2 (IL-2) production by spleen cells in vitro. BEP administration induced a dose-dependent enhancement of the proliferative response to Con A. Infusion of the opiate antagonist naloxone (NAL) inhibited the Con A response and infusion of NAL prior to BEP resulted in even further inhibition. None of these treatments resulted in detectable alterations in IL-2 production after 48 h in culture. To demonstrate a direct interaction between BEP and lymphocytes, spleen cells were incubated in vitro with varying concentrations of BEP and/or NAL. Enhanced Con A-induced proliferation was observed following incubation with BEP in the range 10(-12) to 10(-9) M (levels comparable to the effective in vivo doses) and this effect was abrogated by NAL pretreatment (10(-6) M). These data indicate a role for BEP in enhancing lymphocyte reactivity which is to some extent dependent on opiate receptors on the cell surface. This report extends the evidence obtained from in vitro experiments implicating endogenous opioids in modulation of host immunity by demonstrating that these effects can be obtained in vivo. PMID- 2967723 TI - An efficient algorithm for sequencing peptides using fast atom bombardment mass spectral data. AB - An efficient algorithm is described for sequencing peptides from sequence ions appearing in fast atom bombardment (FAB) and FAB tandem mass spectra. The following features are incorporated in the algorithm. The members of the set of sequence ions are represented by all possible combinations of N- and C-terminal fragment ions. From the known N- and C-terminating groups and molecular weight (MW) of the peptide, the sequence ions are mathematically re-expressed as N terminal residue ions and arranged in ascending order. The peptide sequence is computed, in a stepwise iterative procedure, from the mass differences between the mathematically re-expressed N-terminal residue ions and the predicted peptide subsequences for the neighboring ions of lower mass. These mass differences correspond to combinations of known amino acid residues which have previously been computed and tabulated, based upon the FAB fragmentation rules for peptides. The algorithm was successfully applied to sequence the following peptides from their respective FAB or FAB tandem mass spectrum: decapeptyl (MW 1310), angiotensin II (MW 1045), and two 'unknown' peptides (MW 1227 and 1485, respectively). Two criteria used to predict the correct peptide sequence from among many possibilities are the minimum number of amino acid residues and the maximum fragmentation probability per amino acid residue. PMID- 2967724 TI - Effects of undernutrition during suckling and of novelty on hypothalamic beta endorphin immunoreactivity in young rats. AB - The present report examines the effect of undernutrition and of exposure to an open-field (novelty) on hypothalamic beta-endorphin immunoreactivity in 21-day old rats. The effect of posttraining beta-endorphin administration in open-field performance was also studied. Undernutrition and novelty had no effect on hypothalamic beta-endorphin. Post-training administration of beta-endorphin did not alter the performance of rats in either nutritional group in the test session. The results suggest that the hypothalamic beta-endorphinergic system of young rats is not activated by novelty and that the effect of undernutrition on this system found in adult rats is not detectable at this age. PMID- 2967725 TI - Studies of peripheral blood T lymphocytes in assessment of disease activity in rheumatic fever. AB - The study included three groups of children: (a) 38 with active rheumatic fever (ARF) and active carditis; 21 seen during their first attack and 17 during recurrence of activity, (b) 47 with inactive rheumatic fever (IARF); the period since activity was less than 3 years in 31 cases and more than 3 years in 16 cases. Using monoclonal antibodies and T lymphocyte blast transformation induced by PHA, we found: (1) low total T lymphocytes, helper-inducer cells and helper inducer/suppressor-cytotoxic ratio which persisted for years; and (2) reduced lymphoblast transformation in active disease. PMID- 2967726 TI - Back pain and sciatica. PMID- 2967727 TI - Hoists. PMID- 2967728 TI - Health care of physically handicapped young adults. AB - Little is known about the health needs of physically handicapped young adults after they become too old for the paediatric service. To assess these needs 104 young people with physical handicaps were given a medical examination and interviewed. They had diverse physical handicaps, many of which were rare (41 subjects); 45 had cerebral palsy and 18 spina bifida. Mental handicap (42 subjects) and a need for special education (28) were also diagnosed. The state of health of all subjects was generally poor. Sixty one subjects had contractures of the lower joints and almost a quarter contractures of the upper joints; 26 had deformed feet. Consequently, many subjects (41) could not walk; only 13 could walk without difficulty. Urinary and bowel incontinence were prevalent (58 and 55 subjects, respectively) and with contractures and pressure sores resulted in skin problems (35 subjects). Kyphoscoliosis was present in 30 subjects, particularly those with spina bifida (10 out of 18), of whom one third had respiratory problems and almost half cardiac problems. Thirty one subjects had epilepsy. Many subjects (49) had difficulty communicating or some visual defect (27). Although over half the subjects had health problems that were severe enough to warrant intervention, less than a third were receiving any form of regular hospital care; regular contact with physiotherapists, speech therapists, and dentists was also poor. Moreover, the original diagnoses and prescriptions had not been reviewed regularly. Teams for handicapped adults should be set up in all district health authorities to provide a clinical and coordination service for all adults with physical and mental handicaps. PMID- 2967729 TI - The coexistence of bradykinesia and chorea in Huntington's disease and its implications for theories of basal ganglia control of movement. AB - Investigation of motor function in a group of 17 patients with Huntington's disease reveals that, in addition to the chorea that many patients exhibit, defects in voluntary motor performance also are evident. Fast simple wrist flexion movements to 15 degrees or 60 degrees were slower, and individual movements showed greater variability than seen in normal subjects. This bradykinesia was most pronounced in those patients who were akinetic and rigid, but also was seen in those with chorea alone; bradykinesia was independent of the drug treatment that the patients were receiving (and was therefore not due to drug-induced parkinsonism). The electromyographic activity of the agonist muscles during such simple but slow movement differed from that seen in Parkinson's disease. The performance of complex movements revealed further deficits. Some patients were unable to combine two movements in a simultaneous or sequential movement task of squeezing the hand and flexing the elbow. Those who could perform these complex movements exhibited slowing of the velocity of the movement and prolongation of the interval between movements. These abnormalities were present in patients with chorea who were not taking neuroleptic drugs. It is argued that they represent an abnormality of motor programming of complex movements, over and above the defect in executing simple movements. The long latency stretch reflexes in wrist flexor muscles and flexor pollicis longus were reduced or absent, but this did not correlate with changes in motor performance, or with the reduced size of the early components of cortical sensory evoked potentials. Bradykinesia is thus shown to be an integral component of the motor disorder of Huntington's disease, in addition to the chorea. The coexistence of bradykinesia and chorea in this illness is compatible with current theories of the role of the basal ganglia in the control of movement. PMID- 2967730 TI - [Blood levels of dehydroepiandrosterone sulfate (DHEA-S) and the risk of breast cancer]. AB - In North American women at low or high risk of developing breast cancer, as assessed by an epidemiologic questionnaire, the plasma concentration of dehydroepiandrosterone sulfate shows a statistically significant circannual variation. In adolescents, in all seasons, circulating dehydroepiandrosterone sulfate is a classifier of the risk of developing breast cancer, a relatively low concentration of this hormone being associated with an increased risk. PMID- 2967732 TI - Back pain. AB - Among the multitude of potential etiologies for low back pain in the elderly, cauda equina syndrome is the only genuine surgical emergency. The consequence of delayed recognition, referral, and decompression may be permanent neurologic deficit. Fortunately, this event is rare. Certain other warning signs and symptoms (as outlined in Fig. 1) should alert the clinician to possible systemic disease and warrant earlier, more aggressive evaluation. Careful history-taking and focused physical examination remain the cornerstones of clinical decision making in managing the geriatric patient with back pain. Laboratory and/or radiologic studies are indicated in carefully defined circumstances. Advanced age, independent of other risk factors, is probably not an indication for obtaining roentgenograms and laboratory studies as part of the initial screening of patients with back complaints. Available epidemiologic studies suggest that the predicament as well as the illness of back pain is common in the elderly, and that most people who suffer from back pain in old age had back complaints before the age of 65. Furthermore, although back complaints are among the most commonly mentioned symptoms by elderly patients to their physicians, they are less often the chief or presenting complaint. Studies that examine the reasons for health care-seeking behavior in older patients with back pain, or that compare older and younger patients in this regard, are lacking. Several factors potentially contribute to age-related differences in patient behavior related to back pain. The removal of employment compensation issues in the postretirement population may alter the relative frequency of physician visits. A higher incidence of serious chronic and/or life-threatening disease in the elderly, as well as perceived threats to independent function, may heighten anxiety about back pain. Conversely, because of the greater prevalence of musculoskeletal and other activity-limiting conditions in the elderly, older patients may believe that back pain is just another burden of aging. These questions await further research. PMID- 2967731 TI - The effect of cyclosporin A on Trypanosoma musculi infection of mice. AB - Cyclosporin A treatment of mice infected with Trypanosoma musculi mimicked the effect of T-cell deprivation in increasing the parasitemia and lowering the antibody responses. However, in contrast to T-cell deprivation, cyclosporin A treated mice were still able to clear the parasites from the blood. As expected, T-cell deprived mice were not responsive to cyclosporin A. It is suggested that antibodies against T-independent antigens of the parasite are ineffective in promoting elimination of the trypanosomes. These findings support the importance of T-helper cells in the final elimination of T. musculi from the blood of mice. PMID- 2967733 TI - Rehabilitation management of homebound elderly with locomotor disability. AB - Functional disabilities, the final common pathway of all medical and surgical conditions, are frequently preventable and treatable. Etiologic diagnosis of functional problems is important so that one does not miss the opportunity to improve and increase independence. Rehabilitation of the elderly can be a special problem. Their habits are longstanding and difficult to change. It is important to establish realistic and attainable goals with the patient. Not every habit needs to be altered. What is more important is that the patient safely achieve maximum independence. An environmental assessment is "key" to a full understanding the limitations of function, and how well management strategies can be integrated. PMID- 2967734 TI - 5-Fluoro-2'-deoxyuridine elimination by the isolated perfused rat liver. AB - The influence of dose and hepatic blood flow on the elimination of 5-fluoro-2' deoxyuridine (FdUrd) by the isolated perfused rat liver were investigated. FdUrd (1-20 mg; 4-81 mumol) was injected into the perfusion reservoir and serial samples were collected for chromatographic determination of plasma FdUrd and 5 fluorouracil concentrations. The decrease in FdUrd concentration from values above 100 microM was linear with time (apparent zero order); at concentrations below 30-40 microM the decline became exponential (apparent first order). Semilogarithmic plots of FdUrd concentration/dose versus time obtained with different doses were not superposable, indicating Michaelis-Menten elimination. At a perfusion rate of 20 ml/min, the apparent Vmax and Km for FdUrd disappearance were 14-19 nmol/ml/min and 161-194 microM, respectively. FdUrd clearance during first-order elimination was 8-11 ml/min. After FdUrd administration, 5-fluorouracil concentration reached 10-15% of the initial FdUrd concentration, then decreased with a half-life of 4-7 min. Fifty-four % of the dose of [2-14C]FdUrd was converted to 14CO2. At a dose of 20 mg, first-order clearance of FdUrd increased from 7 to 12 ml/min as hepatic flow increased from 10 to 30 ml/min. Less than 1% of the dose of [6-3H]FdUrd was incorporated into macromolecules. It was concluded that hepatic elimination of FdUrd is dependent on both dose and blood flow. PMID- 2967735 TI - Immunoglobulin and T-cell receptor gene rearrangements in Hodgkin's disease. AB - We have examined tumor tissue DNA obtained from 32 cases of Hodgkin's disease of the following subtypes: lymphocyte predominance, six; nodular sclerosing, eight; mixed cellularity, 14; lymphocyte depleted, 4; using immunoglobulin and T-cell receptor beta and gamma gene probes. Immunoglobulin heavy chain rearrangements were detected in five patients; in three of them only a minor clonal cell population was visible. T-cell receptor gene rearrangement was not observed in any patient examined. Three patients exhibiting minor clonal immunoglobulin rearrangements showed polyclonal T-cells in the same sample. There was no correlation between the presence and intensity of the rearranged bands and the number of Reed-Sternberg cells. Our data do not confirm recent reports of a frequent occurrence of immunoglobulin or T-cell receptor gene rearrangements in Hodgkin's disease and suggest no possible relation between Reed-Sternberg cells and B- or T-lymphocytes, respectively. PMID- 2967736 TI - Comparison of the molecular properties of muscle phosphofructokinase of young and old rats. PMID- 2967737 TI - Evaluation of in vitro activity of teicoplanin against recent clinical isolates. AB - The in vitro antibacterial activity of teicoplanin, a new glycopeptide antibiotic, previously named teichomycin A2, has been compared to that of five other chemoantibiotics: netilmicin, clindamycin, rifampicin, enoxacin and vancomycin. The minimum inhibitory concentration (MIC) values against 588 gram positive strains, 561 facultative aerobes and 27 anaerobes, recently isolated from clinical specimens, were evaluated. Teicoplanin showed the highest activity against all the tested strains, with a geometrical mean of the MICs (GMM), a MIC50 and MIC90 of 0.123, 0.12 and 0.5 micrograms/ml respectively. PMID- 2967738 TI - Myeloprotective effect of high dose medroxyprogesterone acetate (MPA). AB - Results of clinical studies in advanced and, recently, also in early breast cancer have shown a clear protective effect of high dose medroxyprogesterone acetate (MPA) on the bone marrow toxicity induced by chemotherapy. The underlying mechanism of this phenomenon is unsettled and in vitro studies have yielded contradictory results. The results of these clinical and in vitro studies are summarized. The myeloprotective effect may have important therapeutical implications. PMID- 2967739 TI - Heterogeneity of lipoprotein Lp(a) and apolipoprotein(a). AB - We have purified Lp(a) lipoproteins from sera of four subjects by ultracentrifugation, selective precipitation, and chromatofocusing. Each subject had two forms of serum Lp(a) that were separable by chromatofocusing. We purified apolipoprotein (a) [apo(a)] from the eight isolated Lp(a)s and obtained only one form of apo(a) from each subject. The four apo(a)s seen on sodium dodecyl sulfate polyacrylamide gel electrophoresis had different apparent molecular masses, ranging from 275 to 440 kDa. Chemical deglycosylation of the smallest apo(a) yielded a 235 kDa protein, which may be a core protein structure common to all apo(a)s. We conclude that there are many forms of serum Lp(a) and apo(a). The heterogeneity of serum Lp(a) particles can be ascribed in part to differences in size of apo(a), but other factors must account for the existence within a single patient of different Lp(a)s that contain apparently identical apo(a). One must consider the heterogeneity of Lp(a) when designing assays for this lipoprotein. PMID- 2967740 TI - Measurement and characterization of angiotensin peptides in plasma. AB - We report a method for the extraction of angiotensin peptides from plasma with a mixture of acetone, 1 mol/L HCl, and water (40/1/5 by vol). The method is highly reproducible for the measurement of angiotensin I and angiotensin II in small sample volumes, with analytical recoveries of about 80% for both peptides. We investigated the influence of sample handling and found a standard procedure for blood collection, plasma preparation, and extraction was essential. The method was used to measure angiotensin I and II in rat and human plasma. In rat plasma, the mean (+/- SEM) concentrations of angiotensin I and angiotensin II were determined to be 67 (+/- 8) and 14 (+/- 1) pmol/L (n = 10), respectively. Neither angiotensin I nor angiotensin II was detectable 24 h after bilateral nephrectomy. Acute oral administration of the converting-enzyme inhibitor ramipril caused a significant increase of angiotensin I from 85 (+/- 6) to 257 (+/- 33) pmol/L (n = 10; P less than 0.001) and a significant decrease of angiotensin II from 12 (+/- 1) to 7 (+/- 0.4) pmol/L in rat plasma (n = 9; P less than 0.001). In human plasma, angiotensin I and angiotensin II values of 21 (+/- 1) and 6.6 (+/- 0.5) pmol/L (n = 10) were found. A single oral dose of the diuretic furosemide increased angiotensin I significantly from 21 (+/- 1) to 32 (+/- 1.7) pmol/L (n = 5); P less than 0.001), whereas angiotensin II remained unchanged, 6.6 (+/- 0.5) vs 6.4 (+/- 0.4) pmol/L (n = 5). Extracted peptides could be identified as [IIe5] angiotensin I and [IIe5]-angiotensin II by HPLC in combination with specific radioimmunoassays for angiotensin I and angiotensin II. PMID- 2967741 TI - Increased IgA as a predictor of development of AIDS in HIV-infected subjects. PMID- 2967743 TI - General diseases and dental treatability of the institutionalized elderly Finnish population. AB - The purpose of the present study was to investigate the relationship between such general diseases and dental treatability of the elderly living in municipal old people's homes in Finland. The sampling methods used were PPS sampling (probabilities proportionate to size), and non-randomized and randomized sampling. The subjects were 480 residents of 24 municipal old people's homes in different parts of Finland. Of the subjects 153 were men and 327 women, and their ages ranged from 65 to 100 yr. Data on the general diseases was obtained from the subjects' medical histories. The assessment of dental treatability was based on the subjects' communicative ability and cooperation during interview, on a clinical oral examination, and on the subjects' ability to cope with the stress of an interview and examination. Eleven percent of the subjects had no diagnosed diseases, whereas 28% had three or more. The most common diseases in both men and women were cardiovascular diseases followed by respiratory disorders in men and endocrine disorders in women. The dental treatability was considered very poor or poor in 29% and good in 26% of cases. No clear statistical correlation was found between dental treatability and the number of diseases suffered by the subject. However, cardiovascular diseases and neurologic and mental disorders were found to reduce dental treatability more than other disorders. PMID- 2967742 TI - Comparative effects of verapamil and isradipine on steady-state digoxin kinetics. AB - The effects on the steady-state digoxin pharmacokinetics of verapamil (240 mg/day) and a new dihydropyridine calcium channel blocker, isradipine (15 mg/day), were compared. Nineteen healthy white men, aged 23 to 40 years, ingested 0.25 mg digoxin tablets every 12 hours for two consecutive periods of 2 weeks. Each subject also received one of the calcium channel blockers during one of these periods, with agent and sequence randomized. Analyst-blind RIA serum digoxin determinations demonstrated that the nine subjects who received isradipine, 5 mg t.i.d., had a small increment in peak digoxin level from 2.3 +/- 0.6 to 2.9 +/- 0.7 ng/ml (p less than 0.05) but no significant change in steady state level or AUC over 12 hours. By contrast, the 10 subjects who received verapamil, 80 mg t.i.d., showed significant increases in steady-state (0.9 +/- 0.1 to 1.3 +/- 0.2 ng/ml; p less than 0.001) and peak serum digoxin concentrations (2.5 +/- 0.7 to 3.6 +/- 0.8 ng/ml; p less than 0.001) and in AUC (15.7 +/- 1.7 to 23.6 +/- 2.9 ng . hr/ml; p less than 0.001). Neither calcium channel blocker reduced renal digoxin clearance. Verapamil increases digoxin levels without affecting renal clearance. Isradipine has no clinically important interaction with digoxin. PMID- 2967744 TI - Origin of the fourth component of complement related Chido and Rodgers blood group antigens. AB - We have reviewed the relationship between C4 and its related blood group and discussed the mechanisms whereby a fragment of C4 could become attached to erythrocytes (E). We hypothesize that there is chronic fluid-phase activation of C4 by either C1 to form C4b or spontaneous cleavage of the thioester to form iC4. These activated molecules bind to E. Proteolytic degradation of the bound C4b or iC4 would leave a covalently attached fragment of C4 on E and thereby give rise to the Ch and Rg blood group antigens. This system is of further immunopathologic interest since this 'normal' activation or turnover of C4 is closely regulated. In patients deficient in regulatory proteins, this spontaneous or normal turnover of C4 and C3 may initiate a pathologic condition. PMID- 2967745 TI - Occupational contact dermatitis due to glutaraldehyde in health care workers. AB - Allergic contact dermatitis to glutaraldehyde was found in 13 health-care workers with hand dermatitis. Concomitant sensitivity to other chemicals was noted in 10. The positive patch test response was only evident for every 2nd observation in 4. The eruption persisted for more than 6 months in 10 subjects. In 5, the skin disease forced the worker to leave his occupation. PMID- 2967746 TI - Allergic contact dermatitis due to hexamethylenetetramine in core molding. AB - Hexamethylenetetramine is an odorless powder and slowly liberates formaldehyde in an acid medium or when heated. It is usually used in the manufacture of formaldehyde resins, rubber and the explosive, cyclonite. It is also used as a hardener in core molding processes. It was reported as one of the formaldehyde releasers and as a cause of formaldehyde contact dermatitis. We report an allergic contact dermatitis due to hexamethylenetetramine, with a positive patch test reaction to hexamethylenetetramine (1% pet.) but not to formaldehyde (2% aq.). We concluded that this case was an allergic contact dermatitis from hexamethylenetetramine itself. PMID- 2967747 TI - Epidemiology of nickel allergy: results of a follow-up analysis of patients with positive patch tests to nickel. AB - In a previous paper, we reported on nickel sensitivity in 2400 consecutive patients in 5 countries. 3 years later, we reassessed 104 of the 157 nickel positive patients of this former study and found 13 now to be negative to nickel sulphate. Nearly all the others can no longer tolerate nickel contact. 68 patients were free from nickel dermatitis, 16 showed a very mild eczema or dyshidrosis, and 13 were suffering from chronic hand dermatitis. 42 of 104 patients had changed their occupation, 36 successfully, with clearance of skin lesions. Rehabilitation by avoidance of nickel-containing costume jewelry, wrist watches and clothing buckles, and by change of occupation, is possible and necessary. PMID- 2967748 TI - Occupational airborne contact dermatitis in a florist. PMID- 2967749 TI - Contact allergy to iodine in Japanese sargassum. PMID- 2967750 TI - Specific nuclear elimination in polyploid plasmodia of the slime mold Physarum polycephalum. AB - In growing plasmodia of the myxomycete Physarum polycephalum (G2-phase), three distinct classes of nuclei with a relative DNA content of 1x, 2x, and 4x are observed in the presumed haploid strain CL. The 2x and 4x species comprise up to 35% and 5% of the nuclei. Quantitative cytofluorometric studies of nuclei isolated in either G2- or S-phase or after FUDR treatment (G1 arrest) show that the three nuclear populations undergo a synchronous mitotic cycle and that the relative DNA content of the nuclear fractions in G-2 phase reflects the 2c, 4c, and 8c state. The heterogeneity of the nuclear population does, however, seem to be restricted to the growth phase. During a starvation period of 4 days that always preceeds sporulation (and also meiosis), the 4c nuclear population is reduced to 7%, 8c nuclei are no longer detected. These results suggest that a mechanism exists in Physarum for the selective detection and elimination of polyploid nuclei. PMID- 2967751 TI - Changes in trichloroethylene-treated rat erythrocyte membranes in vitro. AB - Trichloroethylene (TCE) is a widely used organic solvent, the most important toxic effect of which is a narcotic central nervous (CNS) effect. In the present study we have used rat erythrocyte membranes as a nerve cell model for studying the changes in membrane integrity caused by TCE treatment. The parameters determined were osmotic resistance and the activities of acetylcholinesterase (AchE) and adenosinetriphosphatase (ATPase), both of which are integral membrane proteins. TCE had a dose-dependent effect on all these parameters. It increased the osmotic resistance at low concentrations and caused a decrease at high concentrations. Enzyme inhibition was only significant at high solvent concentrations. Decrease of temperature potentiated these effects. Our results indicate that changes in membrane proteins may be the initial factor leading to other changes in the membrane, e.g. increased osmotic resistance. PMID- 2967753 TI - The endocrinology of extrapyramidal system disorders. AB - The study of endocrinologic changes in Parkinson's disease, Huntington's disease, and tardive dyskinesia may elucidate the pathophysiology of these disorders, especially the presence of hypothalamic lesions. There is probably a decrease in PRL concentrations in Parkinson's disease, and there may be an increase in TSH response to TRH stimulation. It is not clear if there is any change in GH concentrations in Parkinson's disease. There appears to be a robust increase in GH concentrations in Huntington's disease, and there may be a small increase in PRL as well. At present no endocrinologic abnormality has been well documented in tardive dyskinesia. PMID- 2967752 TI - Regression of left ventricular hypertrophy during combined atenolol and nifedipine treatment. AB - 36 patients with previously untreated essential hypertension and left ventricular hypertrophy were treated with a fixed dose combination of atenolol 50 mg and sustained release nifedipine 20 mg once daily for a mean period of 12.1 months. Echocardiography showed a significant decrease after a mean period of 7.9 months in interventricular septal thickness (13.6%, p less than 0.01), posterior wall thickness (12.6%, p less than 0.001) and left ventricular mass index (18.3%). After 12.1 months the reductions were 20.7% (p less than 0.001), 22.5% (p less than 0.001) and 30.8% (p less than 0.001), respectively. Posterior wall thickness was significantly reduced, but left ventricular end-systolic and end-diastolic dimensions and fractional shortening remained unchanged. Treatment significantly reduced resting blood pressure from 153/105 mm Hg to 122/79 mm Hg (p less than 0.001), and exercise blood pressure at 100W from 189/109 to 157/93 mm Hg (p less than 0.001). Thus, nifedipine in combination with atenolol produces significant blood pressure reduction accompanied by regression of left ventricular hypertrophy without noticeable changes in left ventricular function. PMID- 2967754 TI - Bioassay for A-56268 (TE-031) and identification of its major metabolite, 14 hydroxy-6-O-methyl erythromycin. AB - A-56268 is a new macrolide which is generally two-fold more potent than erythromycin. A new bioassay is described in which plasma samples are extracted with acetonitrile prior to bioassay. The concentration range for the assay is between 0.05-4.0 micrograms/ml, and the concentrations measured are within 6% of those measured by high-power liquid chromatography. An active metabolite which is as active as erythromycin was identified in the plasma. The plasma half-life and area under the plasma curve values of A-56268, as determined by bioassay, were significantly greater than those of erythromycin. PMID- 2967755 TI - Converting enzyme inhibition and the common carotid circulation in older patients with sustained essential hypertension. AB - Arterial diameter, blood flow and vascular resistance of the common carotid artery were studied using a pulsed Doppler system in patients with uncomplicated sustained essential hypertension and compared with age-matched normal subjects. In hypertensive patients below the age of 45 years, arterial diameter and blood flow remained within the normal range, while vascular resistance was increased. In hypertensive patients over 45 years of age, arterial diameter was also normal, but blood flow was reduced and vascular resistance markedly increased. Thus, both small and large arteries were altered in the common carotid circulation of patients with sustained essential hypertension. Acute oral administration of converting enzyme inhibitor produced dilatation of both small (vascular resistance) and large (arterial diameter) arteries in the carotid circulation. In addition, in older patients, blood flow significantly increased and vascular resistance markedly decreased both in acute and long-term situations. Thus converting enzyme inhibition was able to reverse the abnormalities of the common carotid circulation of older subjects with sustained essential hypertension. PMID- 2967756 TI - Neurochemistry of dementia. AB - New neurotransmitter system abnormalities are being described in Alzheimer's disease, Parkinson's disease, and Huntington's disease at a rapid rate. Both classic and neuropeptide systems are affected in cortical and subcortical regions. Comprehensive understanding of the pathophysiology of these disorders will require understanding the multisystem nature and shared features of these disorders. PMID- 2967757 TI - Antigenic heterogeneity of a human melanoma tumor detected by autologous CTL clones. AB - Peripheral blood lymphocytes from a melanoma patient were stimulated with autologous melanoma cells in mixed lymphocyte tumor cultures (MLTC). After three restimulations, the lytic activity of the responder cells directed against the autologous melanoma cells was higher than that against K-562 and autologous Epstein-Barr virus-transformed B cell line (EBV-B) cells. From these MLTC responder cells, we derived specific cytolytic T cell (CTL) clones that lysed the autologous melanoma cells and did not lyse K-562 or autologous EBV-B cells. Autologous melanoma clones were found that were resistant to some or all of these CTL clones. The autologous CTL clones recognized at least two different antigens (A, B) on the melanoma cells and three types of melanoma clones could be distinguished (A+B+, A+B-, A-B-). This antigenic heterogeneity of melanoma clones was confirmed by testing the CTL clones in cold target competition and also in antigen-dependent CTL proliferation assays performed with very small numbers of stimulator cells. The data further indicated an instability of the expression of a melanoma-associated antigen in the course of a long culture period. Among the melanoma clones that expressed antigen A, one was found to stimulate the proliferation of anti-A CTL clones much more effectively than the others. This represents a new type of heterogeneity among tumor cells which may be of significance for the elicitation of an autologous anti-tumoral immune response. PMID- 2967758 TI - Murine Fc gamma receptor proteins: identification of a previously unrecognized molecule with a monoclonal antibody (12-15). AB - Previously we studied differential expression of cell surface molecules between the metastatic murine lymphoma ESb and an adhesion variant ESb-MP. Here we describe the specificity of a monoclonal antibody (12-15) that showed strong binding to the adhesion variant and weak reactivity against ESb cells. The antibody also reacted to lymphoid but not to macrophage-derived cell lines and immunoprecipitated a molecule of approx. 60-69 kDa from ESb-MP cells. N-terminal sequencing of the antigen revealed identity to the beta protein of mouse Fc gamma receptors. Using monoclonal antibodies against Fc gamma receptors (2.4G2 and K9.361) in immunofluorescence assays and cDNA probes specific for alpha, beta 1 and beta 2 Fc receptor transcripts in Northern blot experiments the differential expression of Fc receptors in ESb and ESb-MP cells was confirmed. Biochemical analysis of endoglycosidase F-treated precipitates revealed that antibody 12-15 reacted to products of all three transcripts with molecular masses for the protein core of 38.5 kDa (beta 1), 34 kDa (beta 2) and 31 kDa alpha). In addition, an unknown protein of 37 kDa (termed beta 3) was identified by antibody 12-15 which could also be detected in ESb cells and EL4 cells. Antibodies 2.4G2 and K9.361 did not react to the beta 3 chain but reacted to varying extents to the other Fc proteins in macrophage and lymphoid cells. Comparison by peptide mapping of the novel beta 3 chain to beta 1, beta 2 and alpha proteins revealed similar, but also distinct peptides. The tissue-specific reactivity of monoclonal antibody 12-15 is likely to be due to a carbohydrate epitope associated with all Fc gamma receptors in lymphoid but not macrophage cell lines. PMID- 2967759 TI - Liver-associated macrophage precursor cells proliferate under impairment of regular hemopoiesis. AB - We reported previously that immature macrophage precursor cells can be isolated from spleen and liver of cyclophosphamide or pyran copolymer-pretreated mice. We now extended our investigations to livers of normal, untreated specific pathogen free mice. Using the response to the macrophage growth factor colony-stimulating factor-1 (CSF-1) and the presence of the mouse macrophage-specific F4/80 antigen as criteria of definition, in the liver of normal mice we could demonstrate macrophage precursor (M phi P) cells by means of proliferation assays and flow cytometric analysis. The amount of M phi P present in the normal liver was significantly increased after administration of pyran copolymer. Also an enhanced proliferative response to CSF-1 as well as augmented natural killer activity and cytostasis of Candida albicans was noted in liver nonparenychymal cells (LNPC) after treatment of bone marrow (BM)-irradiated, splenectomized mice with pyran copolymer. Since the irradiated BM was actually proven to be silent by assessment of BM number and proliferative capacity and by scoring white blood cells, our findings suggest a response of endogenous liver M phi P under the applied conditions. Further evidence for the presence of endogenous liver hemopoietic cells was obtained from transplantation experiments in which LNPC brought about the survival of lethally irradiated mice. The data point towards a significance of the liver in disposing hemopoietic cells to the organism under impairment of regular hemopoiesis. PMID- 2967760 TI - The transmembrane orientation of the epsilon chain of the TcR/CD3 complex. AB - The antigen receptor of the T lymphocytes is one of the most complex eukaryotic membrane structures studied to date. The T cell receptor (TcR) consists of two disulfide-linked glycoprotein chains (alpha/beta or gamma/delta) and is noncovalently associated with a group of small and invariable CD 3 proteins. Four CD 3 chains have been recognized: two highly homologous glycoproteins CD 3 gamma and delta, the more distantly related nonglycosylated CD 3 epsilon chain, and the nonglycosylated CD 3 zeta, the latter being present as a homodimer. The unraveling of the architecture of the TcR/CD 3 complex is crucial to our understanding of the processes underlying its assembly, recognition and transmembrane signaling. The transmembrane orientation of the TcR chains and of CD 3 gamma and CD 3 delta can be directly inferred from their primary structure, based on the presence of concensus N-linked glycosylation sites N-terminal of their transmembrane domains. This prediction can not be made, however, for nonglycosylated molecules like the CD 3 epsilon chain. In order to determine the transmembrane orientation of CD 3 epsilon, anti-peptide antisera directed against the N-termini of the human and murine CD 3 epsilon chains were generated in rabbits. Both antisera stained intact T cells, demonstrating that the N-terminus of the CD 3 epsilon chain was located at the outer surface of the plasma membrane. The anti-human CD 3 epsilon peptide antiserum was found to be mitogenic for peripheral blood T cells, a finding previously reported only for monoclonal anti-TcR/CD 3 reagents. Using a novel transient expression system in murine T lymphocytes, the human CD 3 epsilon chain could be expressed on the surface of CD 3+, but not CD 3- murine T cells, as indicated by fluorescence staining with the anti-peptide antiserum. This experiment confirmed the specificity of the anti peptide antiserum and, perhaps more importantly, indicated that the human CD 3 epsilon chain was correctly assembled in the murine CD 3 complex. Moreover, the anti-human CD 3 monoclonal antibody UCHT1 was found to stain T cells expressing the human CD 3 epsilon chain. PMID- 2967761 TI - Macrophages pulsed with sterol-conjugated benzylpenicilloylated eicosa-lysine induce T cell-mediated suppression of IgE antibody formation in mice. AB - The conjugation of two lipophilic p-oxymethylbenzyl-3 beta-cholestanylsuccinate groups to fully benzylpenicilloylated eicosa-L-lysine [BPO20Lys20(OSuco)2] greatly potentiated its ability to suppress anti-BPO IgE antibody formation in vivo in mice. This enhanced tolerogenicity was due to the ability of BPO20Lys20(OSuco)2 to induce T suppressor cells (Luescher, I.F. et al., Eur. J. Immunol. 1984. 14: 68). In the present study the role of adherent cells in the induction of T suppressor cells by BPO20Lys20(OSuco)2 was investigated. Low numbers of macrophages pulsed in vitro with BPO20Lys20(OSuco)2, but not with the hydrophilic analog BPO21Lys20-OH, suppressed anti-BPO IgE antibody formation upon transfer into syngeneic recipients. This suppression was antigen and IgE specific, and was manifested in naive as well as in previously immunized mice. By cell transfer experiments it was found that the suppression was sensitive to cyclophosphamide. The IgE suppression was not due to carry-over of BPO20Lys20(OSuco)2 by the transferred macrophages since the amount of cell associated antigen was approximately 500-fold too low to account for the observed suppressions. The capability of BPO20Lys20(OSuco)2 to elicit anaphylaxis in passively sensitized rats was significantly lower in comparison to the hydrophilic analog BPO21Lys20-OH. Adherent cells pulsed with BPO20Lys20(OSuco)2, although strongly suppressing IgE antibody formation, failed to elicit any detectable anaphylaxis. The role of adherent cells in the induction of T cell mediated suppression of IgE antibody formation by lipid-modified antigens is discussed and novel therapeutical concepts to achieve desensitization of drug allergic individuals are suggested. PMID- 2967762 TI - Role of the 2H4 molecule in the activation of suppressor inducer function. AB - The monoclonal antibody anti-2H4 recognizes a 220-kDa and 200-kDa glycoprotein and subdivides T4+ cells into distinct subpopulations: the T4+2H4+ inducer of suppression and the T4+2H4- inducer of help. The T4+2H4+ subset has been shown to play a crucial role in the activation of T8+ suppressor cells. In the present study, we attempted to determine whether the 2H4 molecule itself may be involved in initial triggering of suppressor inducer function. The results show that the addition of anti-2H4 antibody to a mixture of B and T cells resulted in a marked suppression of pokeweed mitogen-driven Ig synthesis. When anti-2H4 was added to B cell cultures containing T4+2H4+ or T4+2H4- cells but lacking T8 cells, no suppression was generated. In addition to the requirement for T8 cells, no suppression was generated if T4+2H4+ cells were absent. These results suggest that the anti-2H4 antibody contributes to the activation of the T4+2H4+ lymphocyte subset, which in turn induces T8 cells to suppress B cell Ig synthesis. Biochemical analysis of the T4+2H4+ subset of lymphocytes indicated that the in vitro addition of anti-2H4 antibody resulted in an increased expression of the 220-kDa and 200-kDa structure on T4 cells. We conclude that perturbation of the 2H4 molecule may potentiate the activation of the suppressor inducer subset and that the T200 molecule may be directly involved in suppressor inducer function. PMID- 2967763 TI - Interleukin 4 (BSF-1) induces growth in resting murine CD8 T cells triggered via cross-linking of T3 cell surface structures. AB - To analyze the role of interleukin 4 (IL4, BSF-1) during primary activation of resting (high-density) murine CD8 T cells, a model system was used which bypasses antigen-presenting cells by the use of anti-T3 monoclonal antibodies immobilized on Sepharose beads. In high, but not in low cell density cultures, IL4 alone induced cell growth. In low cell density cultures, further to T3 cross-linking a soluble macrophage product was required as co-stimulator to induce sensitivity to IL4. This co-stimulator activity was unrelated to recombinant (r)IL1, rIL6 and rTNF-alpha (tumor necrosis factor alpha). In primary CD8 T cell responses rIL4 driven growth was about half of that induced by rIL2, and not inhibitable by anti IL2 receptor antibodies. Higher concentrations of IL4 down-regulated cell proliferation. In the course of IL4-driven growth, the proliferating cells acquired sensitivity to the growth-promoting effect of IL2. Activated CD4 or CD8 T cells were found to be equally sensitive to the IL4 and IL2-driven growth pathway. Taken together, these results define a physiologic role of IL4 as growth factor during primary activation of resting CD8 T cells and thus extend the spectrum of target cells for IL4. PMID- 2967764 TI - Further evidence for a sequentially ordered activation of T cell rearranging gamma genes during T lymphocyte differentiation. AB - Recently, we have shown that the majority of human peripheral lymphocytes with a T cell receptor (TcR) gamma/delta receptor use a unique gamma chain recognized by the anti-TigammaA monoclonal antibody. This predominantly expressed gamma protein is encoded by a rearranged gene where the V9 segment has joined the JP segment located upstream of the C gamma 1 region (TRGC1). Peripheral TigammaA+ cells were further studied here to shed light on the relation between the two types of TRG rearrangements namely those involving the first (TRG1) vs. the second (TRG2) J-C region. Thirteen clones reactive with anti-TigammaA were tested; it was found that the second TRG allele (i.e., the one which does not involve V9-JP) of these cells was either in germ-line configuration or, more frequently, rearranged with a downstream V gamma gene joined to a J segment of the TRG1 region. These data suggest that productive rearrangements on TRG1 leading to the production of a surface-expressed gamma chain occur before rearrangements involving the TRG2 region. It supports the view that TRG genes are subjected to a sequentially ordered activation during the process of T lymphocyte differentiation. PMID- 2967765 TI - Allele and locus-specific differences in cell surface expression and the association of HLA class I heavy chain with beta 2-microglobulin: differential effects of inhibition of glycosylation on class I subunit association. AB - The assembly of HLA class I antigens, and the contribution of the single N-linked glycan to this process were examined. We observed a requirement for N-linked glycosylation in the proper assembly and surface expression of HLA-B locus products in particular, although considerable variation was seen within the allelic series of the HLA-A and B loci. We conclude that the single N-linked glycan can contribute in a major way to that conformation of the heavy (H) chain which is competent to associate with the light chain beta 2-microglobulin, and that the presence, rather than the type, of carbohydrate chain is important in this respect. The association of human class I H chains with beta 2-microglobulin shows biphasic kinetics, where an initially rapid phase is followed by a prolonged period during which no further association can be measured. It appears that HLA-C H chains are initially synthesized in amounts similar to HLA-A and B H chains, but associate inefficiently with beta 2-microglobulin, resulting in low expression of HLA-C at the cell surface. The individual stages of assembly and maturation of class I antigens including the transfer from Golgi to cell surface were found to display characteristic allelic variation. PMID- 2967766 TI - Anti-CD27 monoclonal antibodies identify two functionally distinct subpopulations within the CD4+ T cell subset. AB - Anti-CD27 monoclonal antibodies react with a cell surface molecule expressed on medullary thymocytes and a large subpopulation (75%) of peripheral blood T lymphocytes. This study was undertaken to analyze the functional capacities of CD27+ and CD27- subpopulations within the CD4+ subset. In addition, we investigated whether CD27 subpopulations belong to two mutually exclusive T cell sublineages. Proliferation upon lectin stimulation by either phytohemagglutinin or pokeweed mitogen (PWM) was found to be consistently higher in CD4+CD27+ cells compared to CD4+CD27- cells. In contrast, CD27+ and CD27- cells did not differ in anti-CD3, soluble antigen or interleukin (IL)2-induced proliferation. In PWM driven B cell differentiation, CD4+CD27+ cells provided helper activity on IgM production, while CD4+CD27- cells did not. The differences observed between CD4+CD27+ and CD4+CD27- cells in both proliferation and T helper activity on IgM production can, at least in part, be explained by the inadequate production of IL2 by CD27- cells upon lectin stimulation. In contrast, CD27+ and CD27- subpopulations did not differ in the production of interferon-gamma. CD27- cells could be induced to express the CD27 antigen through stimulation of these cells with immobilized anti-CD3 antibodies. After 3 days of culture, approximately 50% of the cells had CD27 membrane expression, whereas after 6 days 80% of the cells express the antigen. We conclude that anti-CD27 antibodies identify two functionally distinct T cell populations within the CD4+ subset. These subpopulations apparently do not belong to two separate T cell sublineages, but may reflect differences in the activation state of CD27+ vs. CD27- peripheral blood T lymphocytes. PMID- 2967767 TI - CD45R gives immunofluorescence and transduces signals on mouse T cells. AB - The expression of CD45R on mouse T cells has been studied. This antigen is expressed on the two higher molecular weight bands of CD45 (or T200) and in mouse it is currently used as a marker of B cells (B220). Here we confirm that CD45R is expressed on some mouse T cell clones. We show that a small but measurable proportion of mouse spleen and peripheral blood lymphocyte T cells gives positive immunofluorescence with B220. Also CD45R-specific antibodies increase the proliferation response to phytohemagglutinin up to 3-fold, thus confirming that CD45R molecules transduce a signal into mouse T cells. PMID- 2967768 TI - Recombinant human interferon-gamma induces increased IgE receptor expression on human platelets. AB - Human recombinant interferon-gamma (IFN-gamma) significantly increased the expression of receptors for IgE (Fc epsilon RII) on blood platelets. Fc epsilon RII was measured by specific binding of 125I-labeled IgE or flow cytometry experiments. Scatchard analysis of 125I-labeled IgE binding curves revealed that treatment with IFN-gamma increased the number of Fc epsilon RII but did not change the value of the association constant of Fc epsilon RII for 125I-labeled IgE. IFN-alpha had no effect on the expression or affinity of Fc epsilon RII. In addition to Fc epsilon RII, IFN-gamma also modified the expression of the glycoprotein IIb-IIIa complex on the platelet membrane. PMID- 2967769 TI - Differential effects of pharmacological modulators of cardiac myofibrillar ATPase activity in normal and myopathic (BIO 14.6) hamsters. AB - The effects of four Ca2+ binding protein modulators (bepridil, perhexiline, calmidazolium and trifluoperazine) on cardiac myofibrillar ATPase activity in Triton-purified myofibrils prepared from normal dogs, normal hamsters and age/sex (male, 16-20 weeks)-matched myopathic hamsters (BIO 14.6) have been quantitated. When compared with normal hamsters, myopathic hamster myofibrils have a markedly depressed maximum MgATPase activity (178 +/- 2 vs. 119 +/- 3 nmol/mg per min, respectively) and a slightly increased requirement of Ca2+ for half-maximal activation (ED50; 0.66 vs. 0.75 microM free Ca2+, respectively). Calmidazolium, trifluoperazine and bepridil lower the ED50 for Ca2+ in myopathic myofibrils. Moreover, bepridil and trifluoperazine increase maximum myofibrillar MgATPase activity in myopathic hamster myofibrils. In contrast, calmidazolium depresses maximum and stimulates basal MgATPase activities in myopathic and normal hamster myofibrils. Qualitatively, different effects are apparent when these agents are examined in canine myofibrils. Thus, the pharmacological effects of Ca2+ binding protein modulators on cardiac myofibrillar ATPase activity are dependent upon species and/or pathological state. It is possible to directly enhance a pathological consequence of cardiac myopathy, depression in maximum myofibrillar MgATPase activity, with pharmacological agents. PMID- 2967770 TI - The thromboxane A2/prostaglandin endoperoxide receptor antagonist activity of CV 4151, a thromboxane A2 synthetase inhibitor. AB - The thromboxane A2/prostaglandin endoperoxide (TXA2/PGH2) receptor antagonist activity of CV-4151, a potent TXA2 synthetase inhibitor, was examined. CV-4151 inhibited guinea pig and human platelet aggregation induced by U-44069 with IC50 values of 1.2 +/- 0.3 X 10(-5) and 1.9 +/- 0.4 X 10(-5) M, respectively, and inhibited the specific binding of [3H]U-46619 to washed guinea pig and human platelets with IC50 values of 1.2 +/- 0.3 X 10(-6) and 5.1 +/- 1.0 X 10(-6) M, respectively. CV-4151 competitively inhibited the contraction of rabbit aortic strips induced by U-44069 with a pA2 value of 5.90. In experiments in mice in vivo, CV-4151 (1 and 10 mg/kg i.v.) significantly inhibited the thrombocytopenia induced by U-44069 in a dose-dependent manner. These results show that CV-4151 has a distinct TXA2/PGH2 receptor antagonist effect, and that this effect together with its inhibition of TXA2 synthetase could be important for the pharmacological action of this compound. PMID- 2967771 TI - Antidepressant drugs increase the concentration of neuropeptide Y (NPY)-like immunoreactivity in the rat brain. AB - Rats were given imipramine or zimeldine orally for three weeks and the brain levels of neuropeptide Y-like immunoreactivity (NPY-LI) were measured. The concentration of NPY-LI in frontal cortical tissue was elevated by 40 and 60%, respectively. Imipramine increased the concentration of NPY-LI in the hypothalamus by 65% while zimeldine was without effect. The concentration of NPY LI in the parietal cortex was unaffected by either of the drugs. These observations support the hypothesis that NPY might be involved in the pathophysiology of depressive illness. PMID- 2967772 TI - Cytoplasmic assembly of snRNP particles from stored proteins and newly transcribed snRNA's in L929 mouse fibroblasts. AB - Newly synthesized snRNAs appear transiently in the cytoplasm where they assemble into ribonucleoprotein particles, the snRNP particles, before returning permanently to the interphase nucleus. In this report, bona fide cytoplasmic fractions, prepared by cell enucleation, are used for a quantitative analysis of snRNP assembly in growing mouse fibroblasts. The half-lives and abundances of the snRNP precursors in the cytoplasm and the rates of snRNP assembly are calculated in L929 cells. With the exception of U6, the major snRNAs are stable RNA species; U1 is almost totally stable while U2 has a half-life of about two cell cycles. In contrast, the majority of newly synthesized U6 decays with a half-life of about 15 h. The relative abundances of the newly synthesized snRNA species U1, U2, U3, U4 and U6 in the cytoplasm are determined by Northern hybridization using cloned probes and are approximately 2% of their nuclear abundance. The half-lives of the two major snRNA precursors in the cytoplasm (U1 and U2) are approximately 20 min as determined by labeling to steady state. The relative abundance of the snRNP B protein in the cytoplasm is determined by Western blotting with the Sm class of autoantibodies and is approximately 25% of the nuclear abundance. Kinetic studies, using the Sm antiserum to immunoprecipitate the methionine-labeled snRNP proteins, suggest that the B protein has a half-life of 90 to 120 min in the cytoplasm. These data are discussed and suggest that there is a large pool of more stable snRNP proteins in the cytoplasm available for assembly with the less abundant but more rapidly turning-over snRNAs. PMID- 2967774 TI - Leukaemoid reaction with megakaryocytic features in newborns with Down's syndrome. AB - A leukaemoid reaction was observed in 3 newborns with Down's syndrome. Thrombocytopenia was present in 2, requiring platelets transfusions in 1, and red cell transfusions were necessary in 2 patients. Blast cells characterization by specific monoclonal antibodies showed a prevalence of megakaryoblasts in all 3 cases. This feature was confirmed in 2 of them by the demonstration of platelet peroxidase (PPO) activity under transmission electron microscopy (TEM). A spontaneous remission of the leukaemoid picture was observed after 2-3 months. However, in 1 case a relapse of the myeloproliferative disorder with the same features of the blast cell population was diagnosed after 16 months. Chemotherapy with low-dose Ara-C, started because of a relevant clinical involvement, induced a complete remission. PMID- 2967773 TI - Antagonism of V2-receptor effect of antidiuretic hormone by atrial natriuretic peptide in man. AB - Human alpha-atrial natriuretic peptide (h-alpha ANP) makes the urine of dehydrated volunteers hypotonic to plasma despite high circulating concentrations of antidiuretic hormone. Urinary dilution with h-alpha ANP also occurs in subjects receiving indomethacin. Therefore, h-alpha ANP antagonises effects of antidiuretic hormone on distal tubular V2-receptors in man, probably without involving prostaglandins. PMID- 2967775 TI - Effect of structure of the initiator codon on translation in E. coli. AB - A set of plasmids carrying different initiator codons--either AUG, or GUG, or UUG, or CUG (as a control) in the hybrid gene lacIZ--was constructed by using synthetic oligonucleotides. GUG and UUG codons were demonstrated to be 2-3 times less effective than AUG in translation initiation. Furthermore, the correlation between the efficiencies of different initiator codons in translation initiation proved to vary, depending on the phase of bacterial growth. The rarely occurring usage in nature of the initiator codons GUG and UUG is supposed to be due to the particular role played by the initiator triplets in regulation of gene expression. PMID- 2967776 TI - [Emergency medical care in eye injuries]. PMID- 2967777 TI - [Steroids' action on dependence of outset, maintenance of gravidity and onset of labor]. AB - Dependence of outset, maintenance of gravidity and onset of labor on different kinds of steroid hormones has long been the focus of debate with the underlying control mechanism of these hormones in question remaining left unestablished still in these days. Of author's particular concern in this paper was the two sided aspect of steroid functions. In fact, the steroid may stimulate mother's organ, fetus and placenta to produce much amount of steroid hormone throughout gravidic interval, while at the same time it may control the two contradictory processes, namely the maintenance of gravity and onset of labor through one and the same route. With a view to solve the complexity referred to above an effort has been made through a series of experiments particularity in the term of: 1. Dependent of a system capable to simultaneously determine concentrations of a variety of steroid hormones using the high performance liquid chromatography (HPLC); 2. Profile-analysis to be made on behavior of different kinds of steroids observable over gravidic interval under a support of HPLC system; 3. Behavior of different kinds of hemogenic hormones subsequent to an active DHAS loading in both group with- and without labor pains; 4. Influence of DHAS and gestation on activity of 11 beta hydroxy steroid dehydrogenase in placenta; Some of the results brought by these experiments, which are of some value to speculate on the mechanism of labor-onset and growth of fetus, may be summarized below in line with the preceding items: 1. The measuring system incorporating HPLC made it possible to determine 8 kinds of steroid hormones under systems of C19 and C21 readily in approx. 45 min., even when pretreatment is made involved in the procedure. It was also demonstrably verified that the system shows an acceptable sensitivity combined with a higher correlation with the RIA-method. 2. Profile was successfully delineated of those steroids which would participate in controlling the labor pains inducing mechanism through a series of analytical works made on the behavior of steroid associated with the process of gravidism. With the approach of expected labor, it was seen that 5 kinds of steroid hormones comprising DHA, testosterone, pregnenolone, 20 alpha OH-progesterone, and cortisol showed a marked increase. In contrast, androstenedione and progesterone begin to fall abruptly ever since several days preceding the date of labor. 3. Gain of DHA, 17 alpha OH-progesterone, due to DHAS-loading was clearly identified in the group with onset of labor pains.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967778 TI - Is etretinate dangerous in Papillon-Lefevre syndrome? AB - A 23-year-old girl affected by Papillon-Lefevre syndrome developed, during Etretinate therapy (1 mg/kg/day), two liver abscesses caused by pyogenic bacteria and a subphrenic abscess. The immunological effects of retinoids may precipitate severe infections in patients with Papillon-Lefevre syndrome, who probably have a basic, even though not always evident, defect of PMN chemotaxis. PMID- 2967779 TI - Acitretin (Ro 10-1670, etretin): overall evaluation of clinical studies. AB - Acitretin (Ro 10-1670, etretin) is the free acid derivative of etretinate. This compound possesses a much shorter half-life of elimination than etretinate and was therefore proposed for clinical development. A total of 635 patients with various dermatoses were studied in 12 main clinical trials performed in Europe and in the United States. It was shown that acitretin is effective in the treatment of psoriasis and other diseases of keratinization. Doses between 25 and 35 mg per day are recommended to initiate treatment since marked improvement or clearing was obtained in the majority of patients within this range. Hypervitaminosis A signs and symptoms have been observed in patients treated with acitretin. It is concluded that the efficacy and safety profile of acitretin resembles that of etretinate. PMID- 2967780 TI - A multicentre placebo-controlled study in general practice to evaluate the efficacy and safety of tizanidine in acute low-back pain. AB - Patients (112) with acute low-back pain of recent onset were recruited to this double-blind, randomized, placebo-controlled parallel group study in general practice to evaluate the efficacy and tolerability of the muscle relaxant, tizanidine. They were treated for 7 days with tizanidine (4 mg three times daily) or matching placebo. Aspirin tablets (300 mg) were taken as required as 'rescue' medication. Symptoms were assessed by the patient and doctor before treatment, and after 3 and 7 days. Patients recorded pain and aspirin consumption in a daily diary. Both treatments were effective. In patients who had taken no medication prior to entry, aspirin consumption was almost halved in the first 3 days of taking tizanidine compared with placebo (P = 0.037). Results for pain at rest, pain at night, restriction of movement and pain on movement suggest that tizanidine may give greater improvement, earlier. No serious drug-related adverse events or abnormal biochemistry or haematology were observed in either group. Drowsiness occurred in 22% of patients taking tizanidine although, in patients with severe acute low-back pain, sedation, analgesia and bed rest might be beneficial and desired. Considerably more patients given aspirin/placebo had gastro-intestinal side-effects (P = 0.018). In conclusion, tizanidine may reduce the need for analgesics and be useful in the treatment of acute low-back pain. PMID- 2967781 TI - Tizanidine and ibuprofen in acute low-back pain: results of a double-blind multicentre study in general practice. AB - This study reports on 105 patients with acute low-back pain given tizanidine (4 mg three times daily) plus ibuprofen (400 mg three times daily) or placebo plus ibuprofen (400 mg three times daily). Patients assessed their pain using visual analogue scales in a daily diary and the doctor assessed their condition at baseline and on days 3 and 7. Both groups were treated effectively, but earlier improvement occurred in patients given tizanidine/ibuprofen, particularly regarding pain at night and at rest. Doctors assessed the helpfulness of treatment: tizanidine/ibuprofen was significantly better than placebo/ibuprofen at day 3 (P = 0.05). Significant differences between treatments in favour of tizanidine/ibuprofen occurred in patients with moderate and severe pain at night (P less than 0.05), at rest (P less than 0.05) and those with moderate or severe sciatica (P less than 0.05). Significantly more patients given placebo/ibuprofen had gastro-intestinal side-effects compared with tizanidine/ibuprofen (P = 0.002). This supports previous work in animals showing that tizanidine mediates gastric mucosal protection against anti-inflammatory drugs. More patients given tizanidine/ibuprofen suffered drowsiness and other central nervous system effects (P = 0.025). In patients with severe acute low-back pain, however, some sedation and bed rest is advantageous. This study shows that tizanidine/ibuprofen is more effective in the treatment of moderate or severe acute low-back pain than placebo and ibuprofen alone. PMID- 2967782 TI - Localization and expression of U1 RNA in early mouse embryo development. AB - We have studied the accumulation and localization of U1 RNA during mouse embryo development by in situ hybridization with a U1 RNA probe and immunofluorescence microscopy using a mouse monoclonal antibody to U1 snRNP. There is a substantial amount of U1 RNA present in the oocyte that is present in both the germinal vesicle and the cytoplasm although the concentration is higher in the nuclear compartment. Following the germinal vesicle breakdown that accompanies ovulation and meiotic maturation, the U1 RNA is uniformly distributed throughout the unfertilized oocyte. In the fertilized egg, the silver grain density from in situ hybridization is higher over pronuclei and this enrichment is maintained at the two-cell and later stages. Similar results were obtained for the distribution of the U1 snRNP as assayed by immunofluorescence microscopy: U1 RNA is predominantly localized in all nuclei except polar body nuclei. The U1 RNA in the oocyte and two-cell embryo is predominantly (greater than 85%) U1a RNA. By the eight-cell stage there is a two to three-fold increase in the amount of total U1 RNA and the proportion of U1b RNA has increased to about 40%. The amount of U1 RNA continues to increase through the blastocyst stage and the proportion of the U1b RNA increases to 60%. PMID- 2967783 TI - [Effect of 2 low-dosage Gestodene or desogestrel containing ovulation inhibitors on sex hormones and lipid metabolism]. AB - In a randomised study on 20 healthy female subjects we compared the effect of two low-dosage antiovulants containing gestodene or desogestrel on ovarial and adrenal function and on fatty metabolism. Both antiovulants produced a moderate inhibition of gonadotropin secretion and a marked suppression of serum levels of oestradiol, testosterone and free testosterone; there was a strong increase in sex hormone-binding globulin, and an increase in cortisol--probably due to the increase on corticosteroid-binding globulin--and a continuous drop in dehydroepiandrosterone S. In all cases there was no significant difference between the effect of both preparations. Whereas there was no change in total and LDL cholesterol, total phospholipids and VLDL triglycerides, there was a significant increase in the concentrations of apolipoprotein B, total triglycerides, the cholesterol and phospholipid properties of VLDL as well as of all components of HDL. The LDL triglycerides were also elevated, whereas the LDL phospholipids dropped. On the whole the changes in lipid metabolism, which indicate a certain predominance of the oestrogen effect, were low. However, it became that the duration of intake exercises considerable influence. PMID- 2967784 TI - [Joint pain in pre- and postmenopausal arthropathia climacterica]. AB - The menopausal syndrome consists of a variety of psychic and physical signs of different intensity. At our outpatient department for climacteric complaints and osteoporosis prevention (1. Dept. of Gynecol. and Obstetrics, Vienna) 820 women, who have been consulting this department for the first time because of prevailing complaints or prophylactic reasons, were registered in the year 1986. In the course of this work we paid particular attention to the symptom "pain in a joint" as a sign of the beginning climacteric period. Referring to our patient population we tried to analyse epidemiologically the frequency and the degree of severity of menopausal arthritic pain as well as the therapeutical response to a hormone substitution therapy. To what extent an endocrinologic explanation can be found and which facts let us assume such a statement, we tried to demonstrate in the discussion. PMID- 2967785 TI - [Prenatal sonographic diagnosis of abdominal wall defects and stenoses of the intestinal tract]. AB - Today prenatal sonographic diagnosis of abdominal defects and stenoses of the intestine is possible without few exceptions during general screening in the 20th week of gestation. In a retrospective study, 56 malformations in this area were investigated, which had been diagnosed at the University Hospital Freiburg during 1973 and 1985. Besides the criteria of sonographic diagnosis the etiology, incidence, prognostic factors, the course of pregnancy and delivery and the postpartal management are described. The rate of direct sonographic diagnosis in this area has increased from 26% up to 91%. The worst prognosis was found in infants with diaphragmatic hernias and exomphalos, whereas infants with stenosis of duodenum and small intestine had the best prognosis. PMID- 2967786 TI - Adaptive responses to illness and disability. Healthy denial. AB - The authors attempt to understand the nature and sources of "resilience" by a comparison of three individuals who showed unusual courage in the face of death, illness, and congenital disability. Various adaptive traits that enabled them to function effectively and remain optimistic are discussed. The authors specifically consider the role of "healthy" denial and argue that it does not fully explain or account for their singular optimism confronting adversity. The term "resilience" is proposed to describe both the particular cognitive style of "hardiness" and the "ego strength" that characterize such patients. Possible sources and origins of resilience are suggested. Finally, implications for the treatment of such patients are considered. PMID- 2967787 TI - [Nondisjunction of the chromosomes in man. Hypotheses and facts]. AB - The data on nondisjunction of chromosomes in man, and particularly on chromosome 21 in Down syndrome, are presented in this review. New experimental data obtained by the differential staining technique are considered. These data enable us to revise a number of hypotheses on the causes and mechanisms of chromosome nondisjunction. PMID- 2967788 TI - [Effect of new progressive technological processes in copper-smelting production on working conditions]. PMID- 2967789 TI - [Dust level and microbial contamination of the air in linen spinning and weaving production]. PMID- 2967791 TI - [Improved working conditions for the operators of buffing machines]. PMID- 2967790 TI - [Information value of additional ECG leads in the diagnosis of cor pulmonale in dust-induced bronchitis]. PMID- 2967792 TI - [Means of decreasing occupational diseases of the locomotor apparatus in mica production]. PMID- 2967793 TI - [Effects of pentoxifylline on diastolic heart function in patients with angina pectoris and an increased left ventricular wall mass]. AB - The combination of coronary heart disease (CHD) with increased left ventricular wall mass (LVWM) appears associated with prolonged isovolumetric relaxation (IVR) and consequently, alterations in the rapid filling phase. Methylxanthine substances may improve relaxation through inhibition of phosphodiesterase activity. Accordingly we examined multiple indexes of left ventricular diastolic function before and after administration of 200 mg pentoxifylline (Trental) intravenously to 18 patients (51.3 +/- 9.0 years, 15 males, three females) with stable angina pectoris and positive exercise-ECG in NYHA class I or II and LVWM greater than 160 g (n = 9) and less than or equal to 160 g (n = 9). Left ventricular pressure (P) and volume (V) measurements were made with a high fidelity-micromanometer before and twelve minutes after administration of pentoxifylline. The time constant of left ventricular isovolumic relaxation (T), usual global left ventricular volumes and derived indexes such as peak filling rate (PFR), time to peak filling rate (TPFR), segmental (relaxation and rapid filling phases) and total pressure-volume relationship before and after pentoxifylline were calculated. Significant differences between these two groups (greater than/less than or equal to 160 g LVWM) were found for end-diastolic volume (68.7 +/- 19.0 to 90.8 +/- 22.6 ml/sqm), end-systolic volume (21.7 +/- 16.0 to 36.1 +/- 14.7 ml/sqm), end-diastolic pressure (15.0 +/- 4.8 to 15.7 +/- 5.1 mm Hg), PFR (3.25 +/- 1.18 to 2.66 +/- 0.71 s-1), T (46.0 +/- 5.7 to 52.7 +/- 7.2 ms), the linear regression of lnP-V (lny = -0.117 x + 4.59 to lny = -0.091 x + 4.75) in the IVR-phase (dp/dtmin less than or equal to x less than or equal to 80 ms) (leftward shift in p-V-relationship when less than or equal to 160 g) and the complet p-V-areas. After pentoxifyl-line-administration there were significant decreases in T in patients with increased LVWM (52.7 +/- 7.2 to 47.7 +/- 5.9 ms) and the P-V-product over the time in the rapid filling phase in patients with LVWM less than or equal to 160 g. Total peripheral resistance and heart rate did not change. These changes in parameters of left ventricular diastolic function in combination with significant improvement of pump function especially in patients with LVWM greater than 160 g after administration of pentoxifylline suggest that improved diastolic function is the result of a direct myocardial effect of pentoxifylline. PMID- 2967794 TI - Balloon angioplasty in congenital heart disease. AB - Balloon angioplasty and valvuloplasty offer an alternative to surgery in many congenital cardiac conditions and are a useful adjunct in others. To yield optimal results, the most desirable improvement in hemodynamics with the least amount of damage to normal tissue, choice of the catheter should take into consideration the size of the balloon, type of shaft and shape of the balloon. Experience has shown that dilatation is best achieved when the size of the balloon is such that there is a combination of circumferential force, the stress exerted when the balloon is near its maximal diameter, and longitudinal force, the extent of which is directly proportional to the deformity of the balloon. If the balloon is too small, little benefit may be accrued from the procedure; if it is too large, there is a risk of danger to adjacent normal structures. A stiff shaft helps to maintain a stable position during inflation and the size of the shaft determines the caliber of the guidewire lumen and the inflation/deflation lumen. A single, circular balloon has the advantage of distributing the dilating forces uniformly during inflation; more recently introduced alternative designs with two or three balloons mounted around the shaft have the advantage of allowing some blood flow to occur even at full inflation but they also have the theoretical disadvantage of not ensuring an even distribution of circumferential force. Dilatation of pulmonary valve stenosis may be considered indicated in the presence of a pressure gradient of 40 mm Hg with a right ventricular pressure of 60 mm Hg as the lower limit. In the newborn, this may be qualified by setting the lower limit of right ventricular pressure as 10 mm Hg below systemic arterial pressure if that is 60 mm Hg or less. On choice of the proper balloon size, approximately 20 to 30 percent greater than the pulmonary root, success may be expected in 90% of the cases and, in general, the initial result appears to be that which persists. Although Doppler echocardiography frequently shows pulmonary regurgitation, this is rarely clinically evident. The procedure is safe and only few complications have been reported. Establishing the indication for aortic valvuloplasty usually requires the presence of a systolic gradient of 60 mm Hg without severe regurgitation; mild aortic regurgitation is not a contraindication to the procedure. Reports have indicated good results in both infants and children. This procedure is not without risk and deaths have been reported.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2967795 TI - Chromosomal aberrations and sister chromatid exchanges in lymphocytes and urine mutagenicity of migraine patients: a comparison of chronic feverfew users and matched non-users. AB - 1. Thirty migraine patients who had taken the leaves, tablets or capsules of feverfew daily for more than 11 consecutive months were compared to 30 feverfew non-user migraine patients who had been individually age- and sex-matched. 2. The frequency of chromosomal aberrations and sister chromatid exchanges (SCE) were determined from lymphocyte cultures established from blood samples taken over a period of several months. Matched pairs were sampled on the same date for two thirds of the cases, and the greatest difference in sampling time of the remainder was 20 days. Also, the mutagenicity of urine samples from 10 feverfew user migraine patients was compared to that from 10 matched non-user migraine patients using the Ames Salmonella mutagenicity test system. Paired samples were given on the same date. 3. The mean frequency of chromosomal aberrations in the feverfew user group was lower than that in the non-user group both in terms of cells with breaks (2.13% vs 2.76%) and in terms of cells with all aberrations (4.34% vs 5.11%). However, this difference was small and not significant. 4. The mean frequency of SCE in the feverfew exposed group was lower than that in the control group (8.78 vs 8.80 SCE/cell), but, this difference was not significant as determined by factorial analysis of variance (P = 0.897). There was a highly significant variance between the frequencies of SCE in the matched pairs of migraine patients but this was not related to age, sex or feverfew exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967796 TI - Etodolac overdose. PMID- 2967797 TI - Maternal influence on the immune response: SMLC reactions between identical and reciprocal F1 hybrids and the role of lactation. AB - Identical and reciprocal adult F1 mice from different strain combinations, either nursed on their own mothers or foster-nursed on mothers from the paternal strain, were used to carry out SMLC assays. The results obtained showed that: (1) in vitro proliferation of F1 T cells was significantly different when splenocytes from identical versus reciprocal hybrids were used as the stimulatory population, splenocytes from one of the members of the reciprocal pair being able to induce higher proliferative responses of T cells from both identical and reciprocal F1 hybrids; (2) foster-nursing of F1 hybrids on mothers from the paternal strain was able to induce permanent alterations in the ability of their splenocytes to stimulate the proliferation of responder F1 T cells. The stimulatory ability of splenocytes from foster-nursed hybrids was indistinguishable from that observed in the reciprocal F1 combination nursed by its own mother. The existence of a maternal effect acting through milk on the outcome of self recognition in the litter is discussed. PMID- 2967798 TI - Epidemiological aspects of plasmid profiles in Shigella dysenteriae type 1 strains isolated from Burma & India. PMID- 2967799 TI - Congenital leukemia in an infant with Down syndrome. PMID- 2967800 TI - Use of a glycopeptide antibiotic, teicoplanin, in the treatment of septicaemia caused by gram-positive bacteria. AB - Teicoplanin, a recently introduced glycopeptide antibiotic, has been used, in combination with other antibiotics, to treat 31 episodes of septicaemia caused by Gram-positive organisms. Teicoplanin has double the activity of vancomycin against many Gram-positive bacteria, but allergic reactions and toxicity appear to be infrequent. A single daily dose is sufficient to maintain therapeutic levels, which is an advantage in conditions requiring long-term treatment. Of the 31 episodes treated, 16 were associated with infective endocarditis, 11 with Hickman catheter infection, two with bone and joint infection, and two with infection of other indwelling prosthetic devices. Staphylococcus epidermidis was isolated in 18 infections, of which seven treatment courses were unsuccessful. One death occurred from an uncontrolled infection, three deaths from underlying disease (one of which had relapsed twice), and one after withdrawal of treatment following febrile reaction. Eleven episodes were cured. Six episodes of Staphylococcus aureus septicaemia were treated, of which two failed to respond, two relapsed, one improved and one was cured. The remaining seven episodes were caused by streptococci (including Streptococcus faecalis), and in all of them cure was achieved despite the lack of consistent serum bactericidal activity in vitro. PMID- 2967801 TI - Open study of teicoplanin in gram-positive infections. AB - Eighteen patients with documented Gram-positive infections which included osteomyelitis, prosthetic infections, endocarditis, skin and soft tissue, and urinary tract infections were treated with teicoplanin. The organisms involved included Staphylococcus aureus (15 isolates of which six were methicillin resistant-MRSA), Staphylococcus epidermidis (two), Streptococcus faecalis (one) and Streptococcus milleri (one). Clinical success occurred in all seven patients with skin and soft tissue Streptococcus milleri (one). Clinical success occurred in all seven patients with skin and soft tissue infections (with bacterial persistence in three out of the seven), in three patients with bacteraemia endocarditis, and in one of the three patients with chronic osteomyelitis. In four patients with prosthetic bone and joint infections, clinical improvement followed removal of prostheses. Adverse effects occurred in two patients and these included one patient with a rise in serum aspartate aminotransferase and bilirubin and one patient with a rise in blood urea, both of which returned to within normal limits on discontinuing the drug. The study showed that teicoplanin is a safe and effective antistaphylococcal agent. PMID- 2967802 TI - [Immunopharmacologic principles of drug allergy]. PMID- 2967803 TI - [Hypertension and the heart]. PMID- 2967804 TI - [A 50-year-old patient with laparoscopic biopsy-detected multiple lipid infarcts of the liver and concomitant large, predominantly arterial hemangioma of the liver]. PMID- 2967805 TI - Medical treatment for older people and people with disabilities: 1987 developments. National Legal Center Staff. PMID- 2967806 TI - Euthanasia in The Netherlands. The present day practice of euthanasia. PMID- 2967807 TI - [Propolis-induced contact allergy]. AB - Six patients with contact dermatitis to propolis are reported (3 women, 3 men; mean age 57 years). All three men had eczema on their hands and had acquired their sensitization in the course of bee keeping. The women had been sensitized by propolis contained in ointments of various kinds. Upon patch testing all six patients reacted to propolis and four also to Peruvian balsam. While ten flavonoids were test-negative, a cinnamic acid ester (1.1-dimethylallyl-3',4' dihydroxy cinnamic acid ester) isolated both from propolis and from poplar buds was positive in two out of three patients; this may be the major allergen. PMID- 2967808 TI - [4 cases of Richner-Hanhart syndrome (tyrosinemia type II) with neurological symptomatology in a Yugoslav family]. AB - Findings are described in four cases of Richner-Hanhart syndrome in a Yugoslavian family; this disorder is transmitted as an autosomal recessive trait. This family was first reported in 1963. Now tyrosinemia and tyrosinuria can be found in all cases, and neurological symptoms are also present in all cases, whereas earlier only one of them had these symptoms. In the meantime, the lesions on the soles of the feet have been totally excised in three cases, partially in one, and split thickness grafts from the normal skin have been applied. The palmar lesions were treated in the same manner in two cases. The possibilities for surgical treatment of this very painful, inherited form of dermatosis and their consequences are discussed. Surgical treatment is recommended for palmar lesions, but not for the plantar form. The pathogenesis of the dermatologic manifestations in this rare metabolic disease is briefly discussed. PMID- 2967809 TI - Serogroup specificity and antimicrobial susceptibilities of Neisseria gonorrhoeae isolated in Mexico City. AB - The serogroup pattern of 87 clinical isolates of Neisseria gonorrhoeae was determined by monoclonal coagglutination and the in-vitro activity of seven antimicrobial agents against the same strains was tested by an agar dilution method. The frequency of resistance to spectinomycin, ampicillin, penicillin, erythromycin, chloramphenicol and tetracycline was 14.9%, 33.3%, 34.4%, 30%, 40.2% and 41.3%, respectively. All strains were susceptible to cefotaxime. Out of 87 strains tested, 29.8% produced beta-lactamases and 4.5% were chromosomally resistant to penicillin. In all instances resistance to a drug was associated with serogroup 1-B except for erythromycin. The results presented here correlate with observations made worldwide. PMID- 2967810 TI - In-vitro teicoplanin-resistance in coagulase-negative staphylococci from patients with endocarditis and from a cardiac surgery unit. AB - Among 31 strains of coagulase-negative staphylococcus (CNS) causing endocarditis in individual patients, 16 had MIC of teicoplanin greater than or equal to 8 mg/l (MIC50, 8; MIC90, 8; MIC range, 0.5-32 mg/l); and 24 had MBC greater than or equal to 16 mg/l (MBC50, 32; MBC90, 64; MBC range, 4-128 mg/l). Greater sensitivity was shown to vancomycin (MIC50, 2; MIC90, 4; MIC range, 1-8 mg/l; MBC50, 2; MBC90, 4; MBC range, 0.5-8 mg/l). Teicoplanin-resistant CNS (MIC, greater than or equal to 8 mg/l) were detected in the anterior nares of two of three patients and six of nine staff, and in the air, of a cardiac surgery unit, and in other series of CNS of clinical origin. The results of in-vitro sensitivity testing of CNS to teicoplanin are dependent on the media and conditions used, and their clinical significance has not been determined. Nevertheless, the findings reported here put in question the use of teicoplanin alone as prophylaxis during valve replacement surgery. PMID- 2967811 TI - Activity of teicoplanin against gram-negative anaerobes. PMID- 2967812 TI - Intraventricular administration of teicoplanin in shunt associated ventriculitis caused by methicillin resistant Staphylococcus aureus. PMID- 2967813 TI - Effects of systolic overload and swim training on cardiac mechanics and biochemistry in rats. AB - We have previously shown that swim conditioning corrects the depressed mechanical function and myosin adenosinetriphosphatase (ATPase) activities associated with renovascular hypertension (HTN) in the rat. The present study was designed to assess the effects of swim conditioning on another form of systolic overload, subdiaphragmatic suprarenal aortic stenosis. Cardiac mechanics in an isolated working heart apparatus and myosin enzymology were studied in four groups of rats: controls (C), animals with chronic systolic overload secondary to aortic constriction (St), swim-conditioning animals (Sw), and animals exposed to a combined load (St-Sw). Heart weight was increased by 23% in St, 27% in Sw, and 36% in St-Sw. In contrast to HTN, cardiac pump and muscle function were not depressed in St. Sw was associated with improved cardiac output, stroke work, and velocity of circumferential fiber shortening. St-Sw showed improved mechanical cardiac performance relative to both C and St. The percent of ventricular myosin of the V1 type and Ca2+-activated myosin ATPase activity relative to C was unchanged in Sw but was depressed in St and St-Sw. These data demonstrate that the salutory mechanical effects of Sw can be superimposed on the systolic overload of St. However, the dissociation between mechanics and myosin enzymology suggests that factors in excitation-contraction coupling other than myosin isoenzyme shifts are responsible for this finding. PMID- 2967814 TI - Does a physical handicap alter the development of mastery motivation during the preschool years? PMID- 2967815 TI - Construction of a recombinase-deficient mutant recA protein that retains single stranded DNA-dependent ATPase activity. AB - The recA1 mutation is a single point mutation that replaces glycine 160 of the recA polypeptide with an aspartic acid residue. The mutant recA1 protein has a greatly reduced single-stranded DNA-dependent ATPase activity at pH 7.5 compared to the wild-type protein. Interestingly, the recA1 protein does exhibit a vigorous ATPase activity at pH 6.2. To explore the molecular basis of this pH effect, we used site-directed mutagenesis to replace aspartic acid 160 of the recA1 polypeptide with an isosteric, but nonionizing, asparagine residue. The new [Asn160]recA protein catalyzes ATP hydrolysis at pH 7.5 with the same turnover number as the wild-type protein. This result suggests that the activation of the recA1 protein ATPase activity that occurs at pH 6.2 may be due, in part, to neutralization of the negatively charged aspartic acid 160 side chain. Although it is an active single-stranded DNA-dependent ATPase, the [Asn160]recA protein is unable to complement a recA deletion in vivo and is unable to carry out the three strand exchange reaction in vitro. Further examination of ATP hydrolysis (under strand exchange conditions) revealed that the ATPase activity of the [Asn160]recA protein is strongly suppressed in the presence of Escherichia coli single stranded DNA-binding protein (a component of the strand exchange assay), whereas the ATPase activity of the wild-type recA protein is stimulated by the E. coli protein. To account for these results, we speculate that ATP may induce specific conformational changes in the wild-type recA protein that are essential to the DNA pairing process and that these conformational changes may not occur with the [Asn160]recA protein. PMID- 2967816 TI - Protease Ti, a new ATP-dependent protease in Escherichia coli, contains protein activated ATPase and proteolytic functions in distinct subunits. AB - In addition to protease La (the lon gene product), Escherichia coli contains another ATP-dependent protease, Ti. This enzyme (approximately 340 kDa) is composed of two components, both of which are required for proteolysis. Both have been purified to homogeneity by conventional procedures using [3H]casein as the substrate. The ATP-stabilized component, A, has a subunit molecular weight of 80,000 upon gel electrophoresis in the presence of sodium dodecyl sulfate, but it behaves as a dimer (140 kDa) upon gel filtration. Component P, which is relatively heat stable, is inactivated by diisopropyl fluorophosphate and can be labeled with [3H] diisopropyl fluorophosphate. It has a subunit size of 23 kDa, but the isolated component behaves as a complex (260 kDa) of 10-12 subunits. The isoelectric point of component A is 7.0 and that of P is 8.2, and their amino acid compositions differ considerably. The purified enzyme has an ATPase activity that is stimulated 2-4-fold by casein and other protein substrates but not by nonhydrolyzed proteins. Component A also shows ATPase activity which can be stimulated by casein. Addition of component P (which lacks ATPase activity) inhibits basal ATP hydrolysis by A and makes this ATPase more responsive to casein. Although component P contains the serine active site for proteolysis, it shows no proteolytic activity in the absence of component A, Mg2+, and ATP or dATP. Other nucleoside triphosphates are not hydrolyzed and do not support proteolysis. Protease Ti has a Km for ATP of 210 microM for hydrolysis of both casein and ATP. Casein increases the Vmax for ATP without affecting the Km. A Mg2+ concentration of 5 mM is necessary for half-maximal rates of ATP and casein hydrolysis. Ca2+ and Mn2+ partially support these activities. Thus, protease Ti shares many unusual properties with protease La (e.g. coupled ATP and protein hydrolysis and protein-activated ATPase), but these functions in protease Ti are associated with distinct subunits that modify each other's activities. PMID- 2967817 TI - Radiation inactivation analysis of chloroplast CF0-CF1 ATPase. AB - Radiation inactivation technique was employed to measure the functional size of adenosine triphosphatase of spinach chloroplasts. The functional size for acid base-induced ATP synthesis was 450 +/- 24 kilodaltons; for phenazine methosulfate mediated ATP synthesis, 613 +/- 33 kilodaltons; and for methanol-activated ATP hydrolysis, 280 +/- 14 kilodaltons. The difference (170 +/- 57 kilodaltons) between 450 +/- 24 and 280 +/- 14 kilodaltons is explained to be the molecular mass of proton channel (coupling factor 0) across the thylakoid membrane. Our data suggest that the stoichiometry of subunits I, II, and III of coupling factor 0 is 1:2:15. Ca2+- and Mg2+-ATPase activated by methanol, heat, and trypsin digestion have a similar functional size. However, anions such as SO3(2-) and CO3(2-) increased the molecular mass for both ATPase's (except trypsin-activated Mg2+-ATPase) by 12-30%. Soluble coupling factor 1 has a larger target size than that of membrane-bound. This is interpreted as the cold effect during irradiation. PMID- 2967818 TI - Regulation of embryonic smooth muscle myosin by protein kinase C. AB - Phosphorylation of the 20-kDa light chain regulates adult smooth muscle myosin; phosphorylation by the Ca2+/calmodulin-dependent enzyme myosin light chain kinase stimulates the actomyosin ATPase activity of adult smooth muscle myosin; the simultaneous phosphorylation of a separate site on the 20-kDa light chain by the Ca2+/phospholipid-dependent enzyme protein kinase C attenuates the myosin light chain kinase-induced increase in the actomyosin ATPase activity of adult myosin. Fetal smooth muscle myosin, purified from 12-day-old fertilized chicken eggs, is structurally different from adult smooth muscle myosin. Nevertheless, phosphorylation of a single site on the 20-kDa light chain of fetal myosin by myosin light chain kinase results in stimulation of the actomyosin ATPase activity of this myosin. Protein kinase C, in contrast, phosphorylates three sites on the fetal myosin 20-kDa light chain including a serine or threonine residue on the same peptide phosphorylated by myosin light chain kinase. Interestingly, phosphorylation by protein kinase C stimulates the actomyosin ATPase activity of fetal myosin. Moreover, unlike adult myosin, there is no attenuation of the actomyosin ATPase activity when fetal myosin is simultaneously phosphorylated by myosin light chain kinase and protein kinase C. These data demonstrate, for the first time, the in vitro activation of a smooth muscle myosin by another enzyme besides myosin light chain kinase and raise the possibility of alternate pathways for regulating smooth muscle myosin in vivo. PMID- 2967819 TI - Upstream sequences confer atrial-specific expression on the human atrial natriuretic factor gene. AB - 5'-Flanking sequences from the human atrial natriuretic factor (hANF) gene were subcloned into a reporter plasmid (pSVOCAT) and transfected into primary cultures of neonatal rat atrial cardiocytes. Hybrid hANFCAT genes containing either 2500 or 409 base pairs of 5'-flanking sequence DNA were expressed at similar levels. When sequences between -409 and -332 were deleted, reporter gene (CAT) activity decreased significantly. Expression of the hANFCAT constructs was specific for atrial cells, as no expression was detected in primary cultures of ventricular cardiocytes or nonmyocardial cells derived from the neonatal hearts. Correct transcription start sites for the transfected hANF genes were confirmed by S1 nuclease mapping and RNase protection analysis. A "gel shift" assay was used to identify a specific cardiac nuclear protein which bound to the 5'-flanking sequence of the hANF gene. A 192-base pair PvuII fragment (-400 to -208) associated with a protein in these extracts in a tissue- and sequence-specific fashion. These findings indicate that the DNA sequence between -409 and -332 in the hANF gene harbors a tissue-specific element whose activity may involve association with a cardiac-specific nuclear protein. PMID- 2967820 TI - Immunoglobulin binding by the regular surface array of Aeromonas salmonicida. AB - The cell surface of Aeromonas salmonicida is covered by a regular surface array composed of a single species of protein, the A-protein (Phipps, B. M., Trust, T. J., Ishiguro, E. E., and Kay, W. W. (1983) Biochemistry 22, 2934-2939). The array, known as the A-layer, is the key virulence factor for this organism. Cells containing the A-layer specifically bound rabbit IgG and human IgM with high affinity (KD = 1.0 X 10(-6) M and 3.3 X 10(-6) M, respectively), but neither isogenic A-protein-deficient strains nor an Aeromonas hydrophila strain also possessing a regular surface array had binding activity. Selective removal of A protein at pH 2.2 inactivated IgG binding. Structurally intact IgG was requisite for binding since both Fab and Fc fragments were inactive. Aeromonas A-protein did not share the same IgG binding sites as Staphylococcus aureus protein A. Purified A-protein bound IgG only weakly, but reassembled A-layer regained binding activity. Protein modification and perturbation of the A-layer indicated that no single amino acid residue was critical for binding, and that the binding site consisted of a native arrangement of at least four A-protein monomers in the layer. PMID- 2967821 TI - Biochemical evidence that the type II insulin-like growth factor receptor is identical to the cation-independent mannose 6-phosphate receptor. AB - Cloning and sequencing of the human type II insulin-like growth factor (IGF) receptor cDNA revealed an 80% deduced amino acid sequence homology with the bovine cation-independent mannose 6-phosphate (Man-6-P) receptor, suggesting identity of the two receptors (Morgan, D. O., Edman, J. C., Standring, D. N., Fried, V. A., Smith, M. C., Roth, R. A., and Rutter, W. J. (1987) Nature 329, 301 307). We have performed biochemical experiments that support this proposal. Rat liver type II IGF receptor, purified by the conventional method of IGF-II affinity chromatography, bound quantitatively to a beta-galactosidase affinity column and was eluted with Man-6-P. Bovine liver Man-6-P receptor, prepared by the conventional method of affinity chromatography on phosphomannan-Sepharose, bound IGF-II with high affinity (Kd = 1 nM). Affinity cross-linking of 125I-IGF II to the Man-6-P receptor and analysis by sodium dodecyl sulfate-gel electrophoresis showed that beta-galactosidase, but not Man-6-P, inhibited the formation of the 250-kDa 125I-IGF-II-receptor complex. The inhibition by beta galactosidase was prevented by coincubation with Man-6-P. 125I-IGF-II did not bind to the 46-kDa cation-dependent Man-6-P receptor. For immunologic studies we purified type II IGF receptors and Man-6-P receptors in parallel from rat placental membranes using either IGF-II- or beta-galactosidase affinity chromatography. A panel of five antisera that previously had been raised against either type II IGF receptor or Man-6-P receptor behaved identically toward type II IGF receptor versus Man-6-P receptor in ligand blocking and immunoprecipitation assays. Our data support the conclusion that the type II IGF receptor and the cation-independent Man-6-P receptor are the same protein and that the IGF-II and Man-6-P-binding sites are distinct. PMID- 2967822 TI - Insulin-like growth factor-I is an essential regulator of the differentiation of 3T3-L1 adipocytes. AB - Murine 3T3-L1 preadipocytes proliferate normally in medium containing fetal calf serum depleted of insulin, growth hormone, and insulin-like growth factor-I (IGF I). However, the cells do not differentiate into adipocytes in the presence of the hormone-depleted serum. Supplementation of the growth medium with 10-20 nM IGF-I or 2 microM insulin restores the ability of 3T3-L1 cells to develop into adipocytes. The cells acquire an adipocyte morphology, accumulate triglycerides, and express a 450-fold increase in the activity of the lipogenic enzyme glycerol 3-phosphate dehydrogenase. The increase in glycerol-3-phosphate dehydrogenase activity is paralleled by the accumulation of glycerol-3-phosphate dehydrogenase mRNA and mRNA for the myelin P2-like protein aP2, another marker for fat cell development. IGF-I or insulin-stimulated adipogenesis in 3T3-L1 cells is not dependent on growth hormone. Occupancy of preadipocyte IGF-I receptors by IGF-I (or insulin) is implicated as a central step in the differentiation process. The IGF-I receptor binds insulin with a 70-fold lower affinity than IGF-I, and 30-70 fold higher levels of insulin are required to duplicate the effects of an optimal amount of IGF-I. The effects of 10-20 nM IGF-I are likely to be mediated by high affinity (KD = 5 nM) IGF-I receptors that are expressed at a density of 13,000 sites/preadipocyte. In undifferentiated cells the IGF-I receptor concentration is twice that of the insulin receptor. After adipocyte differentiation is triggered, the number and affinity of IGF-I receptors remain constant while insulin receptor number increases approximately 25-fold as developing adipocytes become responsive to insulin at the level of metabolic regulation. Thus, preadipocytes have the potential for a maximal response to IGF-I, whereas the accumulation of more than 95% of adipocyte insulin receptors and the appearance of responsiveness to insulin are consequences of differentiation. IGF-I or insulin is essential for the induction of a variety of abundant and nonabundant mRNAs characteristic of 3T3-L1 adipocytes. PMID- 2967824 TI - Matrix plasminogen activator inhibitor. Modulation of the extracellular proteolytic environment. AB - We have previously demonstrated that plasminogen activator inhibitor (PAI-1) is associated with the extracellular matrix of cultured bovine smooth muscle cells (Knudsen, B.S., Harpel, P.C., Nachman, R.L. (1987) J. Clin. Invest. 80, 1082 1089). In this report we describe the physiologic role of PAI-1 during the interaction of the tissue plasminogen activator (t-PA) secreting Bowes human melanoma cell line with endothelial extracellular matrices. In addition we have characterized the t-PA.PAI complexes formed during this interaction in the presence and absence of plasminogen. In the absence of plasminogen, a 104-kDa complex between Bowes t-PA and PAI-1 appears in the supernatant. In the presence of plasminogen, PAI initially prevents plasmin formation on the matrix and protects the matrix from degradation by plasmin. The 104-kDa t-PA.PAI complex is degraded into a 68 and a 47-kDa complex by small amounts of plasmin generated from secreted Bowes t-PA and plasminogen. Analysis of these complexes revealed that t-PA is rapidly cleaved by plasmin within the complex whereas complexed PAI 1 is not further degraded. Matrix-associated PAI-1 may play an important role in the protection of extracellular matrices from remodeling and degradation by cellular t-PA and plasminogen. PMID- 2967823 TI - The mechanism of homologous DNA strand exchange catalyzed by the bacteriophage T4 uvsX and gene 32 proteins. AB - A strand exchange reaction between a single-stranded DNA circle and a homologous linear double-stranded DNA molecule is catalyzed by a mixture of two T4 bacteriophage proteins, the uvsX protein (a DNA-dependent ATPase that resembles the recA protein) and the gene 32 protein (a helix-destabilizing protein). The products are different from those formed in the corresponding recA protein catalyzed reaction; rather than producing a linear single strand plus a nicked circular double-stranded (form II) DNA molecule as the final products, interlinked DNA networks are rapidly generated. Electron microscopy reveals that these networks form from multiple pairing reactions that involve the recombination intermediates. Since the uvsX protein is present in substoichiometric quantities, it presumably recycles to catalyze these successive pairing events. Recycling of the uvsX protein has been more directly examined in an assay that monitors the rate of uvsX protein-catalyzed branch migration. The branch migration reaction is rapidly inhibited by dilution of the uvsX protein or by the addition of a heterologous competitor DNA, showing that the uvsX protein DNA filaments that catalyze strand exchange are dynamic structures. The evidence suggests that individual uvsX protein monomers are continuously entering and leaving the cooperatively formed filament in a cycle that is strongly affected by their ATP hydrolysis. PMID- 2967825 TI - Atrioactivase, a specific peptidase in bovine atria for the processing of pro atrial natriuretic factor. Purification and characterization. AB - A seryl protease which catalyzes conversion of proatrial natriuretic factor (ANF) to the active circulating form, ANF(99-126), was purified from a particulate fraction of bovine atria. The enzyme was solubilized with 1.6 M KCl. The molecular mass of the purified enzyme was 580 kDa on gel filtration, whereas by sodium dodecyl sulfate-polyacrylamide gel electrophoresis a cluster of six bands with molecular masses around 30 kDa was observed. The purified enzyme produced ANF(99-126) from partially purified bovine pro-ANF by the selective cleavage of the arginyl peptide bond in the -Pro97-Arg98-Ser99-sequence in pro-ANF. The enzyme was localized mainly in the microsomal fraction rather than the granule fraction. It is likely that the enzyme selectively cleaves the Arg98-Ser99 peptide bond in pro-ANF during the process of secretion. PMID- 2967826 TI - Post-translational protein modification in the endoplasmic reticulum. Demonstration of fatty acylase and deoxymannojirimycin-sensitive alpha mannosidase activities. AB - We have previously described a hybrid protein, GHHA, that contains a fragment of the influenza hemagglutinin joined to the C terminus of a nearly complete rat growth hormone (Rizzolo, L.J., Finidori, J., Gonzalez, A., Arpin, M., Ivanov, I.E., Adesnik, M., and Sabatini, D.D. (1985) J. Cell Biol. 101, 1351-1362). GHHA was transported from the rough endoplasmic reticulum (ER) to a smooth cisterna, continuous with the rough ER, but proximal to the Golgi apparatus. We have now labeled GHHA with [3H]palmitate, demonstrating that fatty acylation can occur in the ER. As expected for a thioester linkage, the label was released from GHHA by hydroxylamine and identified as palmitic acid by thin-layer chromatography. In a second study, we analyzed the structure of the N-linked carbohydrate chain of GHHA. The N-linked oligosaccharides, all high-mannose type, were released by endoglycosidase H and size-fractionated by high pressure liquid chromatography. The predominant structures were Glc1Man8GlcNAc and Man8GlcNAc, indicating that only 2 or 3 glucose and 1 mannose residues were removed from the original Glc3Man9GlcNAc2. Determination of the structure by acetolysis fragmentation indicated that a single Man8GlcNAc isomer was formed by a deoxymannojirimycin sensitive alpha-mannosidase. This contrasts with a previously characterized ER alpha-mannosidase (Bischoff, J., Liscum, L., and Kornfeld, R. (1986) J. Biol. Chem. 261, 4766-4774) that generates the same isomer, but is deoxymannojirimycin resistant. These data suggest the possibility that different enzymes are partitioned within the ER. PMID- 2967827 TI - In vitro stress measurements in the vicinity of six mechanical aortic valves using hot-film anemometry in steady flow. AB - Based on hot-film anemometry, point velocity measurements in the total cross sectional area 1 and 2 diameters downstream of: Bjork-Shiley Standard, Convex Concave and Monostrut, Hall-Kaster (Medtronic-Hall), St. Jude Medical and Starr Edwards Silastic Ball aortic valves were made. The spatial distribution of Reynolds Normal Stresses (RNS) was visualized three-dimensionally in order to point out where and to what extent the highest RNSs were found. The measurements were made in steady flowing glycerol mixture at flow rates 10, 20 and 30 l. min-1 corresponding to mean velocities of 27, 54 and 81 cm s-1. The highest maximum RNS values were around 250 Nm-2 and were found downstream of the Bjork-Shiley Monostrut and Starr-Edwards Ball valves. The lowest maximum RNSs were found downstream of the St. Jude Medical and Hall-Kaster (Medtronic-Hall) valves (125 140 Nm-2). The Starr-Edwards valve had the highest mean RNS (117 Nm-2) followed by the Bjork-Shiley Monostrut (87 Nm-2). These simplified measurements of artificial heart valve performances concerning RNS, enhance the interpretation of results in more complicated flow models not to say in vivo. PMID- 2967828 TI - The association of cystic fibrosis, gastroesophageal reflux, and reduced pulmonary function. AB - Between 1971 and 1984, 57 patients with cystic fibrosis (CF) and clinically suspected gastroesophageal reflux (GER) underwent a barium-meal examination and routine pulmonary function testing. Reflux was demonstrated in 18 patients; in six of these it was complicated by hiatus hernia, esophagitis, or stricture formation. Compared with 412 CF patients without known GER, the 18 patients with demonstrable reflux had significantly reduced forced expiratory volume and forced vital capacity. GER should be looked for carefully on any barium-meal study in patients with CF; these patients have an increased incidence of reflux, with its implications for lung function, and are not good candidates for surgical intervention. PMID- 2967829 TI - Pain in peripheral arteriography: an assessment of conventional versus ionic and non-ionic low-osmolality contrast agents. AB - Two low-osmolality contrast agents, ioxaglate meglumine/sodium and iohexol were compared with diatrizoate meglumine/sodium in a controlled double blind study of 126 patients undergoing arteriography for peripheral vascular disease to determine which caused the least pain. Discomfort was assessed by means of a visual analog scale rating pain from 0 to 100. Average values for pain were 39 +/ 27 for diatrizoate, 14 +/- 15 for ioxaglate and 21 +/- 22 for iohexol. We found that both low-osmolality agents caused significantly less pain in peripheral arteriography than the traditional agent. The p values were p less than 0.0005 for ioxaglate and p less than 0.005 for iohexol versus diatrizoate. In addition, ioxaglate was found to cause significantly less pain than iohexol (p less than 0.05) in this patient group. PMID- 2967830 TI - Metrizamide compared with iohexol for cervical myelography: a double-blind study. AB - A double-blind comparative study of iohexol and metrizamide for use in cervical myelography through lumbar puncture was carried out in a group of 30 patients. No difference was found in the quality of radiographic examinations of the cervical or lumbar regions nor in the postmyelographic computed tomographic examinations of the cervical spine. The overall incidence of side effects was significantly higher in the metrizamide group (93.4%) than in the iohexol group (40%) (p = 0.002). The differences in incidence of nausea (53% in the metrizamide group compared with 13% in the iohexol group) and vomiting (46.6% with metrizamide, 6.6% with iohexol) were also significant (p = 0.02). Pyramidal signs and seizures occurred only after metrizamide myelography. PMID- 2967832 TI - Cardiac transplantation: postoperative chest radiographs. AB - The preoperative and postoperative chest radiographs of 44 patients undergoing 45 orthotopic cardiac allografts were evaluated retrospectively. Patients were followed from two months to four years postoperatively. Radiographic findings were correlated with echocardiograms and endocardial biopsies. The postoperative cardio-pericardial silhouette stabilized in size by six months. Contrary to previous reports, any marked increase in size of the cardiac silhouette in this series was due to pericardial effusion and not to cardiac rejection. Unusual postoperative features of the chest radiographs included calcification of the residual native left atrium, localized pericardial effusion simulating a left ventricular aneurysm, and a prominent left atrial appendage. PMID- 2967831 TI - MRI of periprostatic venous plexus in staging of early prostatic carcinoma. AB - The periprostatic venous plexus can be observed as a bright rim in coronal magnetic resonance (MR) images obtained by spin-echo (SE) 2060/60 technique. A carcinoma of the prostate which penetrates through the capsule into the periprostatic tissues interrupts or obliterates this rim, whereas an intact rim indicates that the tumor is confined within the prostatic capsule. One hundred patients with proven prostatic carcinoma were prospectively imaged by MR to detect periprostatic involvement. The results were compared with those obtained by cystoscopy and digital rectal examination under general anesthesia. The imaging and clinical methods agreed with each other in 76% of the patients. PMID- 2967833 TI - Clinical evaluation of cervical facet joint infiltration. AB - Twenty-one patients with cervical facet syndrome underwent 39 facet joint infiltrations with corticosteroids using fluoroscopic guidance. There were 22 intra-articular and 17 peri-articular injections. This treatment relieved symptoms in 91% of patients. Follow-up ranged from one week to 12 months. However, symptoms usually recurred, specifically in 71% of patients with complete responses and in 42% of those with partial relief. There was no significant difference in response between patients receiving intra-articular or peri articular injections (chi 2 = 2.283). PMID- 2967835 TI - A new direct portosystemic collateral in canine portal hypertension. AB - Two different models of extrahepatic portal obstruction were prepared in dogs to examine the development of portosystemic collateral pathways. The collateral pathways that developed were of two main types, portosplenic communications with the azygos system and a direct communication between the portal vein complex and the intrathoracic part of the inferior vena cava (IVC). This direct communication between the portal venous system and the IVC was unknown to us prior to these studies. The channel we describe has a potential for directly inducing pulmonary embolism, a possibility not seriously considered so far. PMID- 2967834 TI - Percutaneous drainage and aspiration of fluid collections: emphasis on pancreatic collections and transplant patients. AB - Percutaneous aspiration or drainage was the initial treatment of 104 fluid collections in 92 patients. Our success rate was 91%, with a 7% recurrence, 3% complication rate, and 4% mortality rate. Percutaneous drainage (PD) was beneficial in all but two patients, and open surgical drainage was avoided in 84 patients. Included are 18 patients who had pancreatic fluid collections treated, with one drainage failure and one death in this subgroup. Of these patients, 50% had fistulae discovered after drainage, a higher rate than in the patients without pancreatic disease (33%). There are also 10 patients with transplants and subsequent fluid collections, all successfully treated by PD. We conclude that percutaneous abscess drainage techniques are as valuable in the more difficult pancreatic and transplant patients as they are in other patients. PMID- 2967836 TI - Pulmonary blastoma, report of three cases and a review of the literature. AB - Pulmonary blastoma is one of the least common primary malignant tumors of the lung. There is considerable controversy as to its exact histogenesis; it is uncertain if it arises from a single cell or if it is a mixed tumor arising from two different cell lines. We report three patients with pulmonary blastomas, describing in detail their radiological, cytological, and histopathological features. PMID- 2967837 TI - Development of a hospital radiation emergency protocol. AB - Many hospitals are faced with the development of radiation emergency plans. Details are given of the relevant literature and audio-visual materials to assist in the development of such a plan. PMID- 2967838 TI - The radiological report: what is useful for the referring physician? AB - To assess what constitutes a useful radiological report for referring physicians, we sent a questionnaire to 200 doctors (general practitioners, internists, and surgeons). Questions testing style, length of the report, and several points of content, including mention of clinical correlation, negative findings, and sequence of further investigations, were included. The principal qualities useful to the clinician were clarity, brevity, and clinical correlation. Advice on planning of future investigations was especially valued by general practitioners. PMID- 2967839 TI - ICRP Publication 50: lung cancer risk from indoor exposures to radon daughters. PMID- 2967840 TI - Statement from the 1987 Commission Meeting of the International Commission on Radiological Protection. PMID- 2967841 TI - [Chondromyxoid fibroma: radiologic and radioisotope aspects]. AB - Chondromyxoid fibroma is a relatively rare benign bone tumor whose histologic and radiologic patterns are well known. In this article, we describe a 16-year-old boy with such a tumor in his left tibia. Scintigraphically, this tumor is represented by a "doughnut sign". This aspect, though not pathognomonic, has been reported sporadically in a few diseases: osteoporosis circumscripta cranii, angioblastic meningioma, cranial coccidioidomycosis, and aseptic necrosis of frontal bone. Because of their sites, it is easy to reject these diagnoses in our patient. The most pertinent differential diagnosis with regard to the clinical, radiological, and scintigraphic aspects in the patient is that of a giant cell tumor. PMID- 2967842 TI - Ruptured breast implant: computed tomographic and mammographic findings. AB - We report the computed tomographic and mammographic findings in a patient with a ruptured breast implant. The diagnosis was made by recognition of alteration in prosthesis contour, collapse of the prosthesis envelope, and the presence of free silicone in the breast and axillary region. PMID- 2967843 TI - The ultrasonic features of an unusual cholesterol gallstone. AB - We describe a cholesterol gallstone with an unusual macroscopic appearance and only mild acoustic shadowing on ultrasonography with a 3.5 MHz transducer used in vivo; there was greater shadowing using 5.0 and 7.5 Mhz transducters in vitro. PMID- 2967844 TI - The 1987 Gordon Richards memorial lecture: the future of radiography- cassetteless or filmless? AB - A six-year experience of the first cassetteless radiology department has shown that it produces considerable improvement in speed and efficiency of patient handling, reduced physical labor for the technician, and substantial financial savings, amounting to approximately one third of the total non-medical budget. This is contrasted with the potential cost of a totally digital (filmless) department. The digital acquisition devices and their image processing systems are considerably more expensive. Image display comparable to conventional radiographys (for example the full-sized chest film) is markedly inferior in resolution and totally prohibitive in cost. Image storage and recall of the same amount of data as currently handled by the conventional department are presently beyond the capabilities of any commercial communications system, and the potential cost is completely unacceptable. Projections of the imminent advent of the filmless department are grossly over-optimistic, whereas the development of cassetteless systems offers an alternative route to improved patient handling and increased cost-efficiency. PMID- 2967845 TI - Growth disturbances in the hands following thermal injuries in children. 1. Flame burns. AB - Five children developed growth disturbances following severe burns of their hands. Joint abnormalities, amputations, and periosteal reaction were seen on radiographs, and all had premature epiphyseal fusion in several phalanges or metacarpals, a finding rarely described previously. PMID- 2967846 TI - Growth disturbances in the hands following thermal injuries in children. 2. Frostbite. AB - Nine children who had growth abnormalities of their hands following frostbite were studied. In all nine there was shortening of distal phalanges and some adjacent middle phalanges. Proximal phalanges and metacarpals were rarely involved. Epiphyseal abnormalities included destruction, premature fusion, and fragmentation. Other findings included irregularity of the distal ends of phalanges, abnormal alignment, joint abnormalities, and soft tissue swelling. PMID- 2967847 TI - A case against immediate revascularisation after uni- or bilateral removal of infected aortobifemoral (end-to-side) prosthetic grafts. Report based on three cases. AB - More than 75% of the infections of Dacron aortobifemoral grafts occur in the groin. Early removal of the infected limb will enable the vascular surgeon to leave the abdominal part and opposite limb of the graft in place. In aortoiliac occlusive disease and with end-to-side proximal and distal anastomoses, simple removal of the infected graft will not threaten the viability of the limb. Complicated and often incomplete extra-anatomical revascularisation is therefore not necessary and consequently operative time and risk are reduced. Close pre- and postoperative monitoring of the peripheral circulation and painstaking decubitus prophylaxis are mandatory. After 3 to 6 months "in situ" bypass can be performed to correct claudication. Three patients that have been successfully treated according to this philosophy are presented. PMID- 2967848 TI - Autogenous venous reconstruction in the treatment of aortobifemoral prosthetic infection. AB - The Authors describe a patient, in whom an infected aortobifemoral Dacron prosthesis was successfully replaced by an autogenous venous graft. A review of the literature is presented and it is concluded that autogenous reconstruction of the infected field represents a feasible alternative to extra-anatomical revascularisation in the treatment of selected patients with vascular prosthetic infection. PMID- 2967849 TI - Unusual pseudoaneurysm of Dacron femoro-popliteal graft shaft. A case report. AB - An unusual case of pseudoaneurysm in the mid segment of a Dacron femoro-popliteal graft is reported. The radiological diagnosis was based on computed tomography which correlated well with the surgical findings. A manufacturing defect seems to be the most likely cause of this pseudoaneurysm. PMID- 2967850 TI - Gonadotropin-releasing hormone agonists are unsuccessful in reducing tumoral gonadotropin secretion in two patients with gonadotropin-secreting pituitary adenomas. AB - Whether GnRH agonist treatment leads to reduced gonadotropin secretion and tumor volume in patients with gonadotropin-secreting pituitary adenomas is controversial. We studied the effect of GnRH analog treatment in two such patients, one with a recurrent FSH- and LH-secreting pituitary adenoma (patient 1) and one with a recurrent FSH- and alpha-subunit-secreting pituitary adenoma (patient 2). Patient 1 was treated with 200 micrograms Buserelin daily for 65 days, and patient 2 received three injections of 3 mg [D-Trp6]-LHRH formulated in microcapsules at 21-day intervals. In both patients, plasma FSH, LH (RIA), and alpha-subunit concentrations increased initially and remained above the pretreatment values throughout the treatment period. Plasma LH, measured by immunoradiometric assay, remained well above the detection limit. Plasma bioactive LH and testosterone became undetectable in patient 2, but did not change in patient 1. In neither patient did pituitary tumor size (determined by computed tomographic scan) change during treatment. We conclude that 1) the overall effect of GnRH analogs in patients with gonadotroph cell adenomas is stimulation of gonadotropin release by the tumor, although LH release varies according to how plasma LH is measured, possibly related to the origin of the hormone (normal or tumor gonadotroph cells), and 2) GnRH analog treatment does not reduce tumor size. PMID- 2967851 TI - Ontogeny of beta-endorphin and cortisol in the plasma of children and adolescents. AB - The ontogeny of plasma beta-endorphin (B-E) and cortisol concentrations was studied in normal children (3-11 yr old) and adolescents (12-18 yr old) of several ethnic backgrounds. The morning plasma concentrations did not significantly differ as a function of sex or age for either B-E [males (n = 88), 7.19 +/- 0.39 (+/- SE) pmol/L; females (n = 73), 6.80 +/- 0.50 pmol/L] or cortisol [males (n = 87), 383 +/- 14 nmol/L; females (n = 69), 359 +/- 20 nmol/L]. There was a small but significant positive correlation between plasma B E and cortisol concentrations. When sampled immediately after venipuncture, there were no significant differences in the plasma B-E or cortisol concentrations of children who cried during venipuncture compared with those who did not. These data suggest that morning plasma B-E and cortisol concentrations are constant throughout childhood, adolescence, and into early adulthood. PMID- 2967853 TI - Molecular forms of atrial natriuretic peptide in the atrium of patients with cardiovascular disease. AB - To determine the molecular forms of atrial natriuretic peptide (ANP) in patients with cardiovascular disease, we analyzed the ANP content in the right atrium using reverse phase high performance liquid chromatography and RIA. Tissue from 35 patients, ranging in age from 1 month to 64 yr, was studied. The right atrial ANP content was high in patients with congestive heart failure regardless of age. We found three ANP components (alpha-, beta-, and gamma ANP), as previously found in tissue obtained at autopsy. In patients younger than 20 yr old, the major component was gamma ANP. In older patients, the frequency and content of the two low mol wt forms, especially beta ANP, were increased. The differences in the content and molecular forms of ANP in atrial tissue reflect the pathological state in patients with cardiovascular diseases, as well as the patient's age. PMID- 2967852 TI - Binding of proinsulin and proinsulin conversion intermediates to human placental insulin-like growth factor I receptors. AB - Insulin-like growth factor I (IGF-I) and proinsulin share similarities in both primary and tertiary structure. Proinsulin, endogenously secreted or exogenously administered, would, therefore, be expected to interact with IGF-I receptors. We determined the relative activities of IGF-I, insulin, proinsulin, and the proinsulin conversion intermediates in IGF-I radioreceptor assays using term human placental membranes. Insulin was approximately 0.5% as potent as IGF-I, and proinsulin was only 2% as potent as insulin. The six major proinsulin conversion intermediates were studied; all had activities intermediate between those of insulin and proinsulin. We conclude that the binding of proinsulin and the proinsulin conversion intermediates to IGF-I receptors is not of physiological significance at the concentrations occurring endogenously or after exogenous administration of proinsulin. PMID- 2967854 TI - Preliminary investigation of the properties of somatic cell proteases. AB - Effect of the plasmin inhibitor 6-amino-n-hexanoic acid on somatic cell proteases (equivalent to 2.3 X 10(6) cells/ml) was determined using a model system of casein micelles dispersed in Jenness/Koops buffer (1.5% wt/vol). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to quantitate casein. There was no effect of 120 mM 6-amino-n-hexanoic acid on casein proteolysis by somatic cell proteases. Molecular weights of casein proteolysis products produced by somatic cell proteases were different from those produced by plasmin. Quantitative and qualitative analyses of pasteurized mastitic milk by SDS-PAGE indicated that a portion of the somatic cell proteolytic activity survived pasteurization. Because 6-amino-n-hexanoic acid does not inhibit somatic cell proteases, it can be used to establish the relative contribution of somatic cell proteases and plasmin to total proteolytic activity in mastitic milk. PMID- 2967855 TI - Hemodynamic, renal and endocrine effects of atrial natriuretic peptide infusion in severe heart failure. AB - The cardiac release and total body and renal clearances and the hemodynamic, renal and endocrine effects of increasing doses of atrial natriuretic peptide were investigated in 12 patients with severe chronic congestive heart failure. Immunoreactive arterial plasma levels of atrial natriuretic peptide were 10-fold higher than normal and there was no correlation between aortic atrial natriuretic peptide and cardiac filling pressures. The heart released atrial natriuretic peptide into the coronary sinus. The kidney, though a major clearance site, accounted for only 33% of the total body clearance. Administration of 0.3 micrograms/kg per min atrial natriuretic peptide produced significant changes in heart rate (95 +/- 4 to 85 +/- 4 beats/min) and mean arterial (92 +/- 8 to 77 +/- 9 mm Hg), right atrial (13 +/- 3 to 8 +/- 2 mm Hg) and mean pulmonary artery occluded (27 +/- 3 to 14 +/- 3 mm Hg) pressures. Atrial natriuretic peptide increased cardiac index (2.25 +/- 0.18 to 2.83 +/- 0.3 liters/min per m2) and stroke work index (21 +/- 1.5 to 29 +/- 3.4 g/m2), whereas systemic vascular resistance (1,424 +/- 139 to 1,033 +/- 97 dynes.s.cm(-5)) decreased. Infusion of 0.1 microgram/kg per min atrial natriuretic peptide increased urinary flow 128%, fractional excretion of sodium 133% and fractional excretion of potassium 35%. The filtration fraction increased from 29 +/- 2 to 31 +/- 4%. This represented a disproportionate rise in glomerular filtration rate over renal plasma flow. Plasma aldosterone and norepinephrine decreased whereas plasma renin activity remained unchanged. In association with these hemodynamic, excretory and endocrine changes, the urinary excretion of cyclic guanosine monophosphate doubled. Placebo had no effect. These results showed that, despite high circulating levels of atrial natriuretic peptide, administration of this hormone in heart failure is associated with potentially beneficial hemodynamic, renal and endocrine effects. PMID- 2967856 TI - Triple vessel coronary angioplasty: acute outcome and long-term results. AB - Triple vessel coronary angioplasty, defined as angioplasty of one or more lesions in each of the three major coronary arteries (left anterior descending, left circumflex, right coronary artery) was performed in 50 (11%) of 469 patients who had angioplasty of multiple vessels. There were 32 men and 18 women with a mean age of 56 years. All 50 patients had severe three vessel coronary disease and represent approximately 5% of patients with three vessel disease who had revascularization in this institution; 8 (16%) had previous coronary bypass surgery, and 23 (46%) had previous myocardial infarction. Unstable angina was present in 33 patients (66%) and 96% had Canadian Heart Association class III or IV angina; mean left ventricular ejection fraction was 57 +/- 11%. Angioplasty was performed in 176 vessels (3.5 vessels per patient, range 3 to 6) and in 250 lesions (5 lesions per patient, range 3 to 9); angiographic success was achieved in 240 lesions (96%) and 166 vessels (94%). Success in all vessels attempted was achieved in 40 (80%) of the 50 patients. Clinical success (angiographic success associated with clinical improvement) was obtained in all 50 patients in whom triple vessel angioplasty was performed; none of them required urgent bypass surgery and 5 patients (10%) had a non-Q wave myocardial infarction. In four other patients triple vessel angioplasty was planned but not performed because of failure to dilate the primary vessel; urgent bypass surgery was required in one of these, who developed a Q wave infarction. Thus, overall clinical success in 54 patients was 93%; the incidence rate of myocardial infarction was 11%, and that of urgent surgery 1.8%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967857 TI - Restenosis and progression of coronary atherosclerosis after coronary angioplasty. AB - The relation between restenosis and progression of atherosclerosis in other coronary segments after angioplasty was studied in 98 consecutive patients with 110 coronary stenoses successfully treated with angioplasty. At early angiographic restudy (5 +/- 2 months after angioplasty) 37 patients (38%) had restenosis (defined as a stenosis greater than or equal to 50% of the luminal diameter or loss of greater than or equal to 50% of the gain achieved by angioplasty); progression of atherosclerosis was observed in 4 patients with and 7 without restenosis (13 versus 11%, p = NS). Ninety of the 98 patients underwent a late angiographic restudy a mean of 34 +/- 11 months after angioplasty. Late restenosis was found in one patient. Progression of coronary artery disease (defined as a greater than or equal to 20% decrease in the diameter of a vessel initially narrowed by greater than or equal to 50% or a greater than or equal to 30% decrease when the initial stenosis was less than 50%) was examined in relation to restenosis in 85 of the 90 patients. It occurred in 9 of 27 patients with and 22 of 58 patients without restenosis (33 versus 38%, p = NS). Restenosis developed more rapidly than did progression of disease. Diameter stenosis increased from 35 +/- 8 to 73 +/- 11% at the early restudy in lesions with restenosis; in lesions with disease progression it increased from 9 +/- 18 to 20 +/- 28% (p less than 0.001) at the early restudy to 53 +/- 21% (p less than 0.001) at the late restudy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967859 TI - Alternative communication for patients in intensive care. PMID- 2967858 TI - Determination of the catalytic activity of cyclomaltodextrin glucanotransferase by maltotriose-methylorange assay. AB - A kinetic assay of cyclomaltodextrin glucanotransferase (EC 2.4.1.19), based on the use of methylorange dye as the indicator of cyclodextrins, was studied and the reaction verified with independent HPLC analyses. The assay was optimized for the enzyme of an alkalophilic Bacillus sp. (ATCC 21783). Described enzymological data provide a reasonable background for detection of possible errors. Numerical correlations with previous assays are presented. This method can be adopted as a standard analysis of any cyclomaltodextrin-forming enzyme. PMID- 2967860 TI - Haemorheological abnormalities in arterial hypertension and their relation to cardiac hypertrophy. AB - Haemorheological characteristics were measured in a group of 52 patients with essential arterial hypertension (HT), and were compared with those of a group of normotensive subjects. The relationships between the arterial blood pressure (BP), the echocardiographic indices of left ventricular hypertrophy (LVH), and the haemorheological measurements, were studied. The group of hypertensive patients was found to have a hyperviscosity syndrome with significant elevations of blood viscosity at all shear rates (for gamma = 0.20/s, 29.6 +/- 0.6 versus 28.0 +/- 0.3 mPa.s, P less than 0.01: for gamma = 128/s, 4.2 +/- 0.05 versus 4.1 +/- 0.02 mPa.s, P less than 0.02, of plasma viscosity (1.29 +/- 0.01 versus 1.22 +/- 0.06 cSt, P less than 0.001); of erythrocyte aggregation index (17.8 +/- 0.06 versus 14.6 +/- 0.4, P less than 0.001); of erythrocyte filterability index (13.3 +/- 0.5 versus 8.8 +/- 0.2, P less than 0.001) and plasma fibrinogen level (3.4 +/- 0.9 versus 2.8 +/- 0.6 g/l, P less than 0.02). The haematocrit did not differ from that of normotensive subjects (43.3 +/- 0.6 versus 44.7 +/- 0.5%, NS). The left ventricular mass was increased and was positively correlated with the blood viscosity at a high shear rate (r = 0.38, P less than 0.01) and with the erythrocyte aggregation index (r = 0.47, P less than 0.01). Systolic, diastolic, and mean arterial blood pressures were positively correlated with the left ventricular mass (r = 0.34-0.47, P less than 0.05) and with the erythrocyte aggregation index (r = 0.42-0.46, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967861 TI - Forearm haemodynamics in obese normotensive and hypertensive subjects. AB - Forearm haemodynamics using pulsed Doppler flowmetry were studied in 83 men: 15 non-obese and eight obese normotensive subjects; 30 non-obese and 30 obese hypertensive patients. Mean ages were similar in the four subgroups. The blood pressure of normotensives and hypertensives was identical in obese and non-obese subjects. Blood flow, expressed in ml/min, was significantly higher in obese subjects, whether normotensive or hypertensive. However, when blood flow was expressed per unit litre of forearm volume, it was similar in the four subgroups. Forearm vascular resistance, whether expressed as absolute or normalized values, was significantly higher (P less than 0.001) in non-obese hypertensives. When obese and non-obese hypertensives were compared, the former were characterized by higher values of blood flow velocity and blood flow, and lower values of vascular resistance, whether absolute or normalized values were used. This study shows firstly that forearm vascular resistance in hypertensives is increased exclusively in non-obese subjects, and secondly that obese hypertensives, when compared with non-obese hypertensives, are characterized by a hyperkinetic forearm circulation. PMID- 2967862 TI - The effects of atrial natriuretic peptide and amiloride on renal haemodynamics and the renal kallikrein-kinin system. AB - Anaesthetized rabbits were used to examine the effects of amiloride (an inhibitor of a conductive sodium channel in the distal tubule) and atrial natriuretic peptide (ANP), both singly and in combination, on renal function and the renal kallikrein-kinin system. The administration of ANP (0.05 microgram/kg per min) produced a natriuresis with increases in renal blood flow and glomerular filtration rate. The administration of ANP superimposed on amiloride infusion (5 mg/kg + 0.04 mg/kg per min) showed an additive effect on the natriuresis, although the renal haemodynamic changes were now absent. The infusion of ANP alone increased the urinary excretion of kallikrein and kinins. Prior infusion of amiloride prevented the expected increases in the urinary excretion of kallikrein and kinins after infusion of ANP was superimposed. These results suggest that the observed renal haemodynamic changes could be mediated through renal kallikrein and kinins. The additive effect on sodium excretion might be elicited by the results of alterations in the tubular handling of sodium, although distal tubular function is not modified by ANP. It seems that the renal kallikrein-kinin system is not causally involved in the increased sodium excretion by ANP. PMID- 2967863 TI - T helper cell cytoplasmic granules. Exocytosis in response to activation via the T cell receptor. AB - A series of class II MHC-restricted keyhole limpet hemocyanin-specific Th cell clones were examined for cytoplasmic granules by histochemical techniques and fractionation of their homogenates. All showed granules containing lysosomal enzymes and high levels of trypsin-like activity revealed by a N-alpha benzyloxycarbonyl-L-lysine thiobenzyl ester-esterase assay. Using the latter as a marker for granule contents, granule secretion was observed in response to MHC restricted, Ag-dependent signals presented in vitro, and correlated well with T cell activation as measured by proliferation. However, human rIL-2, a strong inducer of T cell proliferation, did not stimulate N-alpha-benzyloxycarbonyl-L lysine thiobenzyl ester-esterase secretion by itself, nor did it influence the secretion mediated by Ag. Other factors found to induce secretion included immobilized antibodies directed against determinants of the TCR complex, as well as Con A. These results suggest that granule enzyme secretion is another indicator of TCR-mediated activation and provides a possible mechanism for Th cell function via a rapid, local delivery of granule contents to adjacent cells. PMID- 2967864 TI - Autoimmune thyroiditis induced in mice depleted of particular T cell subsets. I. Requirement of Lyt-1 dull L3T4 bright normal T cells for the induction of thyroiditis. AB - T cell-depleted C3H/He or (C57BL/6xC3H/He)F1 (B6C3F1) mice were prepared by adult thymectomy and injection of antithymocyte serum, followed 3 wk later by lethal x irradiation and bone marrow reconstitution. When these T cell-depleted mice were not injected or injected i.v. with normal spleen and lymph node cells treated with either anti-Thy-1, -L3T4 or -Lyt-2 antibody plus C or C alone, none of the groups of mice developed thyroiditis. In contrast, the adoptive transfer of normal cells treated with anti-Lyt-1 plus C resulted in high incidence of the production of antithyroglobulin antibody and the induction of typical thyroiditis lesion. The thyroid was the sole organ involved, because neither typical inflammatory lesion in other organs nor autoantibody such as anti-DNA antibody was detected in mice that exhibited thyroiditis. Analyses of surface phenotypes of cells required for inducing thyroiditis by the adoptive transfer revealed that an appreciable percentage of Lyt-1 dull T cells remained after the treatment of normal lymphoid cells with anti-Lyt-1 plus C. Almost all of these Lyt-1 dull T cells expressed magnitudes of L3T4 or Lyt-2 Ag comparable to those detected on Lyt-1 bright T cells. More important, the induction of thyroiditis was almost completely prevented by either in vitro or in vivo elimination of Lyt-1 dull L3T4+(bright) but not of Lyt-1 dull Lyt-2+(bright) T cells. These results indicate that Lyt-1 dull L3T4+ T cells existing in normal healthy individuals have potential to induce typical thyroiditis which is associated with the production of antithyroglobulin autoantibody, and that the activation and/or function of this T cell subset is regulated by the Lyt-1 bright T cell population coexisting in normal lymphoid cell population. PMID- 2967865 TI - Identification of IL-6 as a T cell-derived factor that enhances the proliferative response of thymocytes to IL-4 and phorbol myristate acetate. AB - Thymocytes undergo a vigorous proliferative response when stimulated with a combination of IL-4 and PMA. We have found that conA-induced supernatants from a number of Th cell clones could enhance the level of IL-4/PMA-induced proliferation of unseparated thymocytes 0.5- to 2-fold and of peanut agglutinin positive thymocytes 2- to 10-fold. These supernatants did not contain IL-2 or IFN gamma, and the enhancing activity could be chromatographically separated from IL 3, -4, -5, and granulocyte/macrophage CSF. The possibility that the thymocyte enhancement factor contained in these supernatants was IL-6 was suggested when murine rIL-6 was found to have similar activity. Further evidence for the identity of these two factors was obtained when an IL-6 assay, based on plasmacytoma growth, was used to test column fractions showing thymocyte enhancement. All fractions active in the thymocyte enhancement assay also had activity in the plasmacytoma growth assay. These observations suggest that the thymocyte-stimulating activity present in the T cell supernatants was due to IL 6. PMID- 2967866 TI - Pertussis toxin inhibits human neutrophil responses mediated by the 42-kilodalton IgG Fc receptor. AB - The effects of pertussis toxin (PT) on human neutrophil responses mediated by the 42-kDa IgG Fc R (Fc gamma R42) were compared with its effects on responses mediated by the FMLP receptor. Pre-treatment of neutrophils with PT completely inhibited FMLP stimulation of superoxide production and blocked over 95% of FMLP stimulated degranulation. PT inhibited superoxide production stimulated by Fc gamma R42 cross-linking by 92%. In contrast, degranulation stimulated by Fc gamma R42 was only partially inhibited, with beta-glucuronidase release inhibited by 54%, lysozyme by 33%, and lactoferrin by 78%. With either stimulus, PT inhibition was maximal in the range from 1.8 to 2 micrograms/ml. Responses to both stimuli declined in a parallel fashion with increasing time of exposure to PT with maximal inhibition occurring after 2 h of exposure. Inhibition of FMLP responses and Fc gamma R42-mediated superoxide production, but not degranulation, correlated with ADP-ribosylation of a 45-kDa membrane protein. Inhibition by PT of Fc gamma R42-mediated responses was not due to a change in receptor number. These data suggest that activation of polymorphonuclear neutrophils via Fc gamma R42 proceeds through two pathways, only one of which is regulated by a PT sensitive G protein. PMID- 2967867 TI - IgE formation and Fc receptor-positive lymphocytes in normal, immuno-deficient, and auto-immune mice infected with Nippostrongylus brasiliensis. AB - The IgE serum levels and IgE FcR-positive lymphocytes (Fc epsilon R) in the spleen and mesenteric lymph nodes (MLN) of normal and immunologically mutant strains of mice were determined before and 14 days after infection with Nippostrongylus brasiliensis (Nbr) parasites. By IgE rosetting of cells immunofluorescently stained for sIg. Thy-1.2, Lyt-2, and L3T4, only sIg+ IgE rosetting lymphocytes were detected in both normal and Nbr-infected mice. IgE high responder mice had the same percentage of Fc epsilon R+ spleen and MLN lymphocytes as low responder mice. After Nbr infection, the percentages of splenic and MLN Fc epsilon R+ cells increased in parallel to a similar increase of sIg+ B cells. Athymic C57BL/6J-nu mice had 62% Fc epsilon R+ spleen and 85% Fc epsilon R+ MLN cells before and after Nbr infection, but IgE serum levels were less than 5 ng IgE/ml. C57BL/6J mice with the viable moth-eaten mutation mev which have almost exclusively Ly-1+ B cells, had less than 1% Fc epsilon R+ lymphocytes and formed only small amounts of IgE. C57BL/6J mice with the lymphoproliferation (lpr) or generalized lymphoproliferative disease (gld) mutations had low numbers of Fc epsilon R+ cells but formed 15 to 30 times more IgE after Nbr infection than control C57BL/6J mice. The IgE response of mice with the beige mutation (bg) did not differ from control mice. Mice with the xid mutation had few Fc epsilon R+ and sIg+ cells but showed high IgE responses. These data demonstrate that Fc epsilon R are typical cell surface markers for approximately 90% of murine Ly-1-, sIg+ B cells and that the number of Fc epsilon R+ cells does not correlate with the capacity of the mice to form IgE. The IgE response to Nbr infection is normal in mice homozygous for the bg mutation, elevated in mice homozygous for the xid, lpr, and gld mutations, and decreased in mice homozygous for the mev and nu mutations. PMID- 2967868 TI - Laparoscopic terminal contraception in camp approach. PMID- 2967869 TI - We believe--in organ transplantation. PMID- 2967870 TI - Nursing care plans in the ambulatory surgical eye center. PMID- 2967871 TI - Marketing a healthcare facility. PMID- 2967872 TI - Liability insurance is essential to professional security. PMID- 2967873 TI - Research: a process essential to nursing practice. PMID- 2967874 TI - Postfusional latency in stereoscopic slant perception and the primitives of stereopsis. AB - Random dot stereograms of slanted surfaces were constructed, each representing one or two slanted surfaces in different relative arrangements and with different axes. Latency to fusion and from fusion to stereoscopic resolution was measured for each stimulus. It was found that latency to fusion was always very brief but that latency to stereoscopic resolution varied markedly, depending upon the orientation and arrangement of the stereoscopic surfaces. A gradient of discontinuities at a surface boundary produced an instant slant response for that surface, whereas a gradient of absolute disparities across the surface did not, except under conditions where vertical declination (a form of orientation disparity) was present. We conclude that stereopsis is not based on the primitives used in matching the images for fusion and that it is, at least initially, a response to disparity discontinuities which play no role in the fusion process. We also conclude that vertical declination is responded to globally as a slant around a horizontal axis but that other forms of orientation disparity are ineffective. The evidence from our experiments does not support the existence of a stereoscopic ability to respond globally to differences in magnification (or spatial frequency). It is suggested that stereoscopic perception of slant around a vertical axis is slow because it results from the integration of local processes. PMID- 2967875 TI - Multitask investigation of individual differences in hemispheric asymmetry. AB - Right-handed subjects (N = 120) participated in four different laterality tasks designed to measure aspects of cerebral hemisphere asymmetry: identification of dichotically presented consonant-vowel syllables (CVs), examination of the effects of concurrent repetition of CVs and concurrent anagram solution on finger tapping by the right and left hands, lateralized identification of CVs presented tachistoscopically to the left and right visual fields, and left/right biases on a free-vision face task involving judgments of emotion. Ear differences in the dichotic listening task were related to the pattern of lateralized interference in the dual-task finger-tapping paradigm. There were no other significant relations between pairs of tasks, but when the present results are considered in the light of other recent experiments, there appears to be a relation between lateral bias on the free-vision face task and visual field differences in tachistoscopic identification. The pattern of results has implications for hypothesized individual differences among right-handers in cerebral dominance for verbal processes, input pathway dominance, and asymmetric arousal of the two cerebral hemispheres. PMID- 2967876 TI - Expectancy and visual-spatial attention: effects on perceptual quality. AB - When we expect important stimuli at a particular spatial location, how does our perceptual sensitivity change over space? Subjects were cued to expect a target stimulus at one location and then required to perform one of the following tasks at that and three other locations: luminance detection, brightness discrimination, orientation discrimination, or form discrimination. The analysis of subjects' performance according to signal detection theory revealed changes in both sensitivity and bias for each of these tasks. Sensitivity was maximally enhanced at the location where a target stimulus was expected and generally decreased with distance from that location. Factors that influenced the gradient of sensitivity were (a) the type of task performed and (b) the spatial distribution of the stimuli. Sensitivity fell off more steeply over distance for orientation and form discrimination than for luminance detection and brightness discrimination. In addition, it fell off more steeply when stimuli were near each other than when they were farther apart. PMID- 2967877 TI - Priming effects between two-dimensional shapes. AB - The role of reference frames in shape perception was examined by investigating priming effects between two-dimensional shapes. Ambiguous shapes and shapes with an unambiguous axis were used, and primes and targets could be orientationally transformed so that for ambiguous shapes, different axes aligned with the vertical. Reliable priming effects were established that were dependent on the structural similarity of primes and targets, the informativeness of the prime, and the interval between the stimuli. Most interestingly, with short interstimulus intervals, differential priming effects were found between ambiguous shapes and unambiguous shapes when an orientational transformation aligned different axes in the ambiguous shape with the vertical. The result was interpreted as demonstrating that priming was determined by the similarity of the frame-based descriptions of primes and targets. The finding occurred irrespective of the informativeness of the prime, suggesting that it reflects obligatory processes in shape perception. Further data showed that under some circumstances prespecifying the orientation of a shape also facilitated its processing; this provided direct evidence that shapes are represented relative to a perceptual reference frame. PMID- 2967878 TI - S-R compatibility between response position and destination of apparent motion: evidence of the detection of affordances. AB - In choice reaction time, stimuli and responses in some combinations (e.g., based on spatial arrangement) are faster than in other combinations. To test whether motion toward a position yields faster responses at that position, a computer generated square in front of one hand appeared to move either toward that hand or toward the other hand. Compatible responses (e.g., motion toward left hand/left response) were faster than incompatible responses, even when that opposed traditional positional compatibility. In Experiment 2, subjects responded to the same stimuli but with both hands left, right, or on the body midline. Medial responses were the fastest, showing that destination, rather than mere relative position, was a critical variable. It was suggested that spatial compatibility effects are not unique to position but apply to a variety of task situations, describable by J.J. Gibson's theory of affordances, in which he claims that one perceives the actions (e.g., catching) permitted in a situation. PMID- 2967879 TI - Understanding the central processing limit in consistent-mapping visual search tasks. AB - Effects of load (i.e., the number of stimuli in the display) have been observed in multiple-frame studies using a consistent mapping of stimuli to responses (e.g., Fisher, 1982, 1984). In a series of four experiments, it is shown that these effects are not the consequence of differences across the high- and low load conditions in either decision noise or peripheral masking. Additionally, it is shown that of two modes of limited capacity (a limited-channel and divided capacity model) considered as possible explanations of load effects in tasks where subjects are required to locate a target, only one--the limited-channel model--is consistent with the results from all three location tasks. Finally, it is argued that the limited-channel model predicts not only the behavior observed in the four consistent-mapping experiments reported in this article but also the behavior observed in several related consistent-mapping tasks (Kleiss & Lane, 1986; Shiffrin & Gardner, 1972). PMID- 2967880 TI - Effects of aging and task difficulty on divided attention performance. AB - We report two experiments that compare the performance of young and older adults on perceptual-motor tasks involving division of attention. Previous studies have shown older people to be especially penalized by divided attention situations, but the generality of this finding was recently challenged by Somberg and Salthouse (1982). The present study was conducted to investigate the possibility that age differences in dual-task performance are amplified by an increase in the difficulty of the constituent tasks, where difficulty was manipulated by varying the central, cognitive nature of the tasks (Experiment 1) or the degree of choice involved (Experiment 2). With the present tasks, strong evidence was found for an age-related decrement in divided attention performance. Contrary to our original expectations, however, it does not seem that division of attention presents some especial difficulty to older people. Rather, division of attention is one of several equivalent ways to increase overall task complexity. In turn, age differences are exaggerated as tasks are made more complex. PMID- 2967881 TI - The distorted room illusion, equivalent configurations, and the specificity of static optic arrays. AB - The distorted room illusion (DRI) and the attendant argument for perceptual ambiguity is critically analyzed from a Gibsonian/ecological point of view. The notions of multiple specification, conflicting information, and perceptual skill are invoked in showing how the ecological approach can accommodate illusion effects that may remain under mobile binocular viewing conditions. Static optic arrays are shown not to be ambiguous. So-called equivalent configurations are found to be analytic artifacts, appearing when the problem of information is treated in geometrical terms without regard for constraints due to physical and ecological regularities. The relative importance of motion-based and motion independent information is discussed. PMID- 2967882 TI - Affine distortions of pictorial space: some predictions for Goldstein (1987) that La Gournerie (1859) might have made. AB - Goldstein (1987) studied the perception of pictures seen from the front and the side. Several distinctions arose from his results and analysis, but only one is central to the reanalysis presented here: The perceived orientation of objects within a picture with respect to the external world is a function of viewer position in front of the picture. For example, the eyes of a portrait subject appear to follow an observer who moves around a gallery. Viewed from many positions, such objects can be said to rotate, following a mobile viewer. Goldstein called this the differential rotation effect because those objects that point directly out of the picture (at 90 degrees) rotate most; those pointing at other angles rotate in decreasing amounts. Goldstein offered no theoretical model and little in the way of explanation for this effect. This Observation offers a model based on the affine geometry and the analyses of La Gournerie (1859). This analysis transforms pictorial space (the space behind a photograph or representational picture) by shears, compressions, and dilations according to the viewpoint of the observer in relation to the composition point of the picture. These effects account for Goldstein's differential rotation effect quite well. PMID- 2967883 TI - Geometry or not geometry? Perceived orientation and spatial layout in pictures viewed at an angle. AB - Cutting (1988) suggests that changes in the perceived orientations of pictured objects that occur with changes in viewing angle are caused by the geometrical changes that accompany these changes in viewing angle. His geometrical analysis does predict the differential rotation effect reported by Goldstein (1979, 1987), but fails to predict other important aspects of the data. Cutting's analysis does, however, support Goldstein's (1987) conclusion that in future research on picture perception it is important to clearly distinguish between the attributes of perceived orientation and spatial layout. PMID- 2967884 TI - Space is to time as vision is to audition: seductive but misleading. AB - Space and time serve two perceptual functions. First, space/time forms a framework for visual and auditory events. Second, spatial and temporal change defines the properties of events and objects. It is at this second level that correspondences (i.e., mappings) between visual and auditory qualities can be hypothesized. Due to the active nature of perceiving, all such mappings illustrate the possible relations between looking and listening. PMID- 2967886 TI - [Robertsonian translocation and genetic counseling]. AB - A collaborative study on 92 Robertsonian translocations is analysed in relation with the methods of ascertainment, the type of rearrangement and potential imbalance of the anomaly. The results are useful in genetic counselling. PMID- 2967887 TI - [Chromosome translocations: study of 232 cases originating from 144 families]. AB - Between 1974 and 1987, 232 translocation carriers have been detected in our Center; they belong to 144 different families. Indications for chromosome analysis were the following: familial studies in relation with a patient suggesting a chromosome anomaly (25.4%); mental retardation with or without malformations (24.6%); 2 or more spontaneous abortions (17.2%); infertility problems, mainly male (16.4%); genetic counseling for a non-chromosomal disease (9.5%); prenatal diagnosis in risk pregnancies (6.9%). The chromosome anomalies detected were the following; balanced Robertsonian fusions (114 cases = 49.1%); balanced translocations (74 cases = 31.9%); unbalanced translocations, Robertsonian fusions included (44 cases = 19%). Two groups may be distinguished: the first one confirms data already known, such as high frequency of balanced translocations in couples with multiple abortions, or in infertile males. The second group on the contrary shows more unusual observations: 4 cases of standard trisomy 21 born to young parents carriers of a balanced translocation not involving chromosome 21; 5 cases of trisomy 13 with 46 chromosomes and a Robertsonian fusion, born to parents carriers of a t(13q; Dq) (twice the mother and thrice the father); 14 cases of apparently balanced translocations, however with an abnormal phenotype; and finally 22 cases of balanced translocations incidentally detected during the course of investigations in patients with a genetic problem generally not associated with a chromosome defect. PMID- 2967885 TI - Muscle force and stiffness during activation and relaxation. Implications for the actomyosin ATPase. AB - Isolated skinned frog skeletal muscle fibers were activated (increasing [Ca2+]) and then relaxed (decreasing [Ca2+]) with solution changes, and muscle force and stiffness were recorded during the steady state. To investigate the actomyosin cycle, the biochemical species were changed (lowering [MgATP] and elevating [H2PO4-]) to populate different states in the actomyosin ATPase cycle. In solutions with 200 microM [MgATP], compared with physiological [MgATP], the slope of the plot of relative steady state muscle force vs. stiffness was decreased. At low [MgATP], cross-bridge dissociation from actin should be reduced, increasing the population of the last cross-bridge state before dissociation. These data imply that the last cross-bridge state before dissociation could be an attached low-force-producing or non-force-producing state. In solutions with 10 mM total Pi, compared to normal levels of MgATP, the maximally activated muscle force was reduced more than muscle stiffness, and the slope of the plot of relative steady state muscle force vs. stiffness was reduced. Assuming that in elevated Pi, Pi release from the cross-bridge is reversed, the state(s) before Pi release would be populated. These data are consistent with the conclusion that the cross bridges are strongly bound to actin before Pi release. In addition, if Ca2+ activates the ATPase by allowing for the strong attachment of the myosin to actin in an A.M.ADP.Pi state, it could do so before Pi release. The calcium sensitivity of muscle force and stiffness in solutions with 4 mM [MgATP] was bracketed by that measured in solutions with 200 microM [MgATP], where muscle force and stiffness were more sensitive to calcium, and 10 mM total Pi, where muscle force and stiffness were less sensitive to calcium. The changes in calcium sensitivity were explained using a model in which force-producing and rigor cross-bridges can affect Ca2+ binding or promote the attachment of other cross-bridges to alter calcium sensitivity. PMID- 2967888 TI - Bovine chromaffin cells have insulin-like growth factor-I (IGF-I) receptors: IGF I enhances catecholamine secretion. AB - The binding of 125I-insulin-like growth factor-I (125I-IGF-I) to bovine chromaffin cells was measured. Chromaffin cell cultures contained 111,000 +/- 40,000 IGF-I binding sites/cell. These sites bound IGF-I with a KD of 1.1 +/- 0.3 nM and had a much lower affinity for insulin. Cross-linking studies showed that 125I-IGF-I bound to a protein that had an Mr of approximately 125,000, similar to the Mr of the alpha subunit of the IGF-I receptor in other tissues. Cells cultured with IGF-I (10 nM) for 4 days exhibited an almost twofold increase in high K+-evoked catecholamine secretion. Insulin was much less potent than IGF-I in enhancing catecholamine secretion. These data indicate that binding of IGF-I to its receptors on chromaffin cells can modulate the function of these cells. PMID- 2967889 TI - Beta-thromboglobulin in cerebral infarction. AB - Plasma beta-thromboglobulin (BTG) was significantly elevated in the acute phase of 116 atherosclerotic thrombotic (p less than 0.0001) and 36 cardioembolic (p less than 0.005) infarcts but normal for 96 lacunes compared with controls. This elevation persisted into the 6th week after the acute event. Among atherosclerotic thrombotic infarcts, the acute beta-thromboglobulin level showed a tendency to correlate with infarct size on CT and predicted mortality at 6 weeks. These results suggest that platelet aggregation plays a primary role in the pathogenesis of atherothrombosis. PMID- 2967890 TI - Increased platelet aggregation and release reaction in myotonic dystrophy. AB - Platelet aggregation (PA) induced by (-)-epinephrine and adenosine diphosphate (ADP) was studied in 16 patients with myotonic dystrophy (MyD) and 14 healthy subjects. Plasma beta-thromboglobulin level (beta-TG), a useful marker of in vivo platelet release reaction, as well as in vitro 5-[14C]hydroxytryptamine (5-HT) release, were also studied. The extent of PA induced by (-)-epinephrine at 1 and 3 min and by ADP at 3 min was significantly higher in the patients than in controls. Plasma beta-TG and ADP- or epinephrine-induced platelet 5-HT release were also increased in MyD patients. These results suggest that an abnormality in release as well as in alpha 2-receptor functioning occurs in the platelets of MyD patients. The relation of this abnormality to changes in Ca2+ fluxes through the platelet membrane is discussed. PMID- 2967891 TI - Clinical features of the failed-back syndrome. AB - This study comprises 78 patients who were treated for chronic back pain at a multidisciplinary, multimodal pain treatment center. These patients were selected from 494 patients examined by the authors because all of their previous medical records, operative notes, and imaging studies were available for review. The records and imaging studies were reviewed independently by a neurosurgeon and an orthopedist, and a retrospective decision was made concerning the historical and physical findings correlated with imaging studies in order to provide justification for the intervention. At the time of admission to the pain treatment center, 16 patients had no physical abnormalities that would explain their back complaint and 16 patients exhibited minor postoperative changes insufficient to cause disabling pain. Twenty-seven patients suffered from a complication of previous surgery, 13 had spondylotic disease, and in six a new diagnosis was established. Comprehensive psychiatric evaluation of the 78 patients revealed that 10 patients had a definitive psychiatric diagnosis, 34 were diagnosed as having a maladaptive personality disorder, and 34 had a normal pre-pain personality. Sixty-seven patients suffered from reactive depression. Fifty-four patients were taking medications at doses higher than usually prescribed, 58 misused narcotics, nine had drug addiction, and 54 were suffering withdrawal symptoms. Of the 78 patients, 64 underwent a total of 171 operations, an average of 2.6 per patient. The authors applied the clinical criteria approved by the American Association of Neurological Surgeons and the American Academy of Orthopedic Surgeons for selection of surgery or chemonucleolysis in the treatment of the herniated disc to these patients. Preoperative imaging studies were normal or demonstrated nonspecific degenerative disc disease in 52 patients. Twenty-six patients had a diagnosis based on radiological findings that warranted surgery. Clinical criteria justifying intervention were met in 25 patients and not met in 53. Imaging and clinical criteria for a second operation were met in 18 (40%) of the patients. After the second operation all patients met the criteria: subsequent surgery was necessary to treat effects of an earlier operation in 73%. These data indicate that many of these patients with failed-back syndrome underwent an original operation based on a persistent complaint of pain, frequently coupled with an underlying psychiatric abnormality, although they did not meet the criteria generally accepted by neurosurgeons for intervention at the time of first surgery. PMID- 2967892 TI - Handicap discrimination considerations in treating the impaired worker: drugs, alcohol, pregnancy, and AIDS in the workplace. AB - In dealing with applicants and employees, administrators must comply with handicap and pregnancy discrimination statutes, any union collective bargaining agreement, and any state or local statutes specifically directed at protecting certain classes of handicapped persons. The employer must make reasonable accommodations which allow the disabled worker to perform the job, although the employer need not implement accommodations which are an undue hardship. Similarly, if the impaired employee cannot safely perform the essential functions of the job, he or she need not be hired or retained. Employers must also recognize their responsibilities and liability under their health benefit plans and other liability statutes, and attempt to minimize their exposure. PMID- 2967893 TI - Cancer as a cause of back pain: frequency, clinical presentation, and diagnostic strategies. AB - Back pain is very common. Rarely, it may be the first manifestation of cancer. Although many advocate selective use of laboratory and x-ray tests for back pain patients, the early detection of cancer may be an important reason to obtain such tests. To develop a diagnostic approach that would identify malignancies while remaining parsimonious, the authors evaluated 1,975 walk-in patients with a chief complaint of back pain. Thirteen patients (0.66%) proved to have underlying cancer. Findings significantly associated with underlying cancer (p less than 0.05) were: age greater than or equal to 50 years, previous history of cancer, duration of pain greater than 1 month, failure to improve with conservative therapy, elevated erythrocyte sedimentation rate (ESR), and anemia. Combining historical features and ESR results led to an algorithm that would have limited x ray utilization to just 22% of subjects while recommending an x-ray for every cancer patient. It would further suggest which patients with negative x-ray findings require further work-up. PMID- 2967894 TI - Gloves and the practice of dentistry. Part 3. Recent developments concerning AIDS. PMID- 2967895 TI - Residual muscular swelling after repetitive eccentric contractions. AB - This investigation illustrates the morphological changes that take place following eccentric exercise and correlates those changes with intramuscular pressure readings. Eight healthy male subjects were asked to exercise their right lower leg anterior compartment eccentrically and their left concentrically. Four hundred submaximal contractions were performed in each exercise regimen over a 20 min period against a load corresponding to 15% of the individual's maximal dorsiflexion torque. Tissue fluid pressures were measured by the slit catheter technique before, during, and after exercise and 48 h later. Needle biopsies of both anterior tibialis muscles were also taken 48 h after completion of the exercise regimens. Overall morphology of the specimens revealed a greater cross sectional fiber area (both type 1 and type 2) in the eccentrically exercised muscle as compared with the concentrically exercised muscle. Scant evidence of inflammation (only 1 of 8 of the "eccentric" muscle samples) and no fiber necrosis was observed. Fiber type proportions were equal on both sides and type 1 fiber biased (70%). Extremely large type 2 fibers were found in 4 of 8 subjects from the eccentric specimens. This incidence correlated significantly with the length of the time to return to resting pressure after eccentric exercise (r = 0.93, p less than 0.001). The percentage of water content was significantly higher in the eccentrically exercised muscle. Based on these findings, we conclude that muscle fiber swelling is a predominant feature following eccentric exercise and is directly associated with delayed muscle soreness. PMID- 2967896 TI - Autonomic arousal feedback and emotional experience: evidence from the spinal cord injured. AB - We interviewed spinal-cord-injured, other handicapped, and nonhandicapped subjects to investigate the relation between the perception of autonomic arousal and experienced emotion. The three groups differed significantly on only one measure of affect intensity, with the spinal-cord-injured subjects more often reporting stronger fear in their lives now compared with the past. In addition, spinal-cord-injured subjects often described intense emotional experiences. Spinal-cord-injured subjects who differed in their level of autonomic feedback differed in intensity on several measures. Subjects with greater autonomic feedback tended to report more intense levels of negative emotions. The findings indicate that the perception of autonomic arousal may not be necessary for emotional experience. There were weak trends in our data, however, suggesting that the perception of arousal may enhance the experience of emotional intensity. The subjective well-being reports of the handicapped groups were comparable to those of nonhandicapped subjects, indicating successful coping with their disability. PMID- 2967897 TI - [The relationship between the levels of trace elements in the serum of mice bearing subcutaneous tumors and the use of laser, hematoporphyrin derivatives, gamma-ray and hyperthermia treatment]. PMID- 2967898 TI - Health education strategies for tropical disease control in school children. AB - This study was concerned with the demonstration of the outcome of health education in the control of malaria, schistosomiasis, dracontiasis and onchocerciasis in rural secondary schools in Nigeria using a simplified version of Green's antecedent model. Green identified three factors which could contribute to the prevalence of a disease or health problem and they are 'predisposing', 'enabling' and 'reinforcing' factors. Thus educational activities were planned in this project that would affect these three factors. The study was of a quasinon equivalent before and after experimental design. It was carried out in 10 secondary schools grouped into two equal clusters (five each) to form experimental and control groups. The post-intervention findings revealed that there was a significant increase in the knowledge of the four diseases among those in the experimental group over those in the control group. The results also showed that when knowledge was supported by enabling and reinforcing factors, desirable changes could result in the school setting among students. PMID- 2967899 TI - Steroid content of the peripheral and adrenal vein in Cushing's syndrome due to adrenocortical adenoma and carcinoma. AB - Selective adrenal vein catheterization was done preoperatively in 11 patients with Cushing's syndrome owing to either an adrenal adenoma (6) or an adrenal carcinoma (5). Peripheral and adrenal venous blood (tumor side) was analyzed for progesterone, 17-hydroxyprogesterone, testosterone, delta-4-androstenedione, dehydroepiandrosterone, dehydroepiandrosterone sulfate, cortisol and 11 deoxycortisol. Comparisons were made of the ratios of precursors to products in the androgen and cortisol production pathways. In adrenal vein samples, cortisol levels were higher in the adenoma patients (p equals 0.06). These patients had a higher ratio of 17-hydroxyprogesterone to delta-4-androstenedione (p less than 0.04). The ratio of 11-deoxycortisol to cortisol was higher in the carcinoma patients (p less than 0.02). In the peripheral samples, the carcinoma patients had higher levels of testosterone and 11-deoxycortisol (p less than 0.02) and progesterone (p equals 0.05). The most striking differences were in the peripheral levels of 17-hydroxyprogesterone, delta-4-androstenedione and dehydroepiandrosterone, which were 10 to 20 times higher in the carcinoma group (p less than 0.005). The ratios of progesterone to 17-hydroxyprogesterone and 17 hydroxyprogesterone to 11-deoxycortisol were higher in the adenoma group (p less than 0.005 and p equals 0.06, respectively). The peripheral blood ratio of 11 deoxycortisol to cortisol was higher in the carcinoma group (p less than 0.03). These data indicate that peripheral and adrenal venous steroid levels and ratios of precursors to products are significantly different in the 2 groups. The carcinomas have a block in the conversion of 11-deoxycortisol to cortisol with a subsequent accumulation in androgen and cortisol precursors. The elevations in 17 hydroxyprogesterone, delta-4-androstenedione and dehydroepiandrosterone, and in the ratio of 11-deoxycortisol to cortisol in the carcinoma group are most striking in the periphery. Peripheral blood steroid measurement in patients with Cushing's syndrome and an adrenal mass aids in the preoperative differentiation of carcinoma from adenoma. PMID- 2967900 TI - [Successful transluminal coronary angioplasty in a patient with ischemic heart disease with severe stenocardia and Wolff-Parkinson-White syndrome]. PMID- 2967901 TI - [Comparative characteristics of serum lipids in healthy persons and in patients with coronary arteriosclerosis living in low and medium altitude areas of Tadzhikistan]. AB - Clinical symptoms, atherogenic (cholesterol, triglycerides, beta-lipoproteins + pre-beta-lipoproteins, low-density lipoprotein cholesterol, very low density lipoprotein cholesterol, atherogenic coefficient) as well as antiatherogenic (alpha-lipoproteins, high density lipoprotein cholesterol and its proportion in total cholesterol) lipoproteins, and phenotypes were examined in 727 individuals (330 normal subjects and 397 atherosclerotic patients), living in two different areas of Tadzhikistan (830 and 2200 m above sea-level). In midaltitude dwellers coronary atherosclerosis was associated with low incidence of clinical and electrocardiographic coronary signs and a comparatively favorable clinical course, apparently due to low atherogenic and high antiatherogenic lipid parameters, the absence of type-4-hyperlipoproteinemia and low incidence of type 2B hyperlipoproteinemia. PMID- 2967902 TI - [The C-cell system of the thyroid in rats following a flight on the Kosmos 1667 biosatellite]. AB - Histological, electron-microscopic and morphometric investigations of the thyroid gland of Wistar SPF male rats (aged 3 months) flown for 7 days on Cosmos-1667 showed that its parenchyma was functionally active and changed but little as compared to the controls. However, at an acute stage of adaptation to microgravity C-cells showed morphological signs of their functional decline: the number of low activity cells and cells whose cytoplasm contained secretory granules increased, the volume of nuclei decreased significantly (by 16.2% as compared to the control), and dystrophic changes seen ultrastructurally appeared. These observations together with the results obtained in prolonged animal flights suggest that in microgravity the synthesis and excretion of the hormone calcitonin diminish. In combination with other factors, the functional decline of C-cells inhibits bone neoformation and enhances bone resorption. PMID- 2967904 TI - [Histological study of the lymphoid organs of rats after a 7-day space flight on the Kosmos 1667 biosatellite]. PMID- 2967903 TI - [Water and electrolyte content of the organs and tissues of male rats following a flight on the Kosmos 1667 biosatellite]. AB - After the 7-day space flight onboard the biosatellite Cosmos-1667 the water, Na, K, Ca and Mg content of the liver, kidney, heart, skin and bone of male rats was measured. No significant changes in the weight or water content of the above organs were seen. The exception was a decrease of water contained in the heart and an increase of water contained in the caudal appendage of the epididymis. After flight the mineral composition of the liver was identical to that after control studies. The K content of the heart of the flight rats was lower and that of Na, Ca and Mg was identical to the parameter in the controls. The K content of the skin and bone increased and the Na content of the skin also grew. In the kidney the Ca content did not change whereas the content of K, Na and Mg decreased significantly. In the testis Na decreased and K increased after flight. Thus, changes in fluid-electrolyte homeostasis at the organ and tissue level can develop within 7 days of space flight. They occur not only in the musculoskeletal system but may also evolve in the nonweight-bearing organs. PMID- 2967905 TI - Influence of cardiac flow rate on turbulent shear stress from a prosthetic heart valve. AB - Elevated turbulent shear stresses associated with sufficient exposure times are potentially damaging to blood constituents. Since these conditions can be induced by mechanical heart valves, the objectives of this study were to locate the maximum turbulent shear stress in both space and time and to determine how the maximum turbulent shear stress depends on the cardiac flow rate in a pulsatile flow downstream of a tilting disk valve. Two-component, simultaneous, correlated laser velocimeter measurements were recorded at four different axial locations and three different flow rates in a straight tube model of the aorta. All velocity data were ensemble averaged within a 15 ms time window located at approximately peak systolic flow over more than 300 cycles. Shear stresses as high as 992 dynes/cm2 were found 0.92 tube diameters downstream of the monostrut, disk valve. The maximum turbulent shear stress was found to scale with flow rate to the 0.72 power. A repeatable starting vortex was shed from the disk at the beginning of each cycle. PMID- 2967906 TI - [The mesoduodenum]. AB - In the present study an anatomical examination of ten bodies was made to describe the fixation of duodenum and pancreas to the posterior abdominal wall. It has become apparent that the shape of the posterior mesoduodenum of the first part of duodenum is rodlike. Due to the secondary retroperitoneal fixation between duodenum and pancreas a vascularized, wide-based borderline lamella remains in the area of second and third parts of the duodenum. The borderline lamella corresponds to the tela subserosa and strengthens Treitz fascia and Gerota fascia in the prerenal area. It covers the inferior vena cava, and is fixed to the adventitia of the abdominal aorta. The retroduodenopancreatical tissue forms a duodenopancreatical layer which contains its own vessels. Now it is possible to understand better the surgical importance of pancreatitis, retroduodenal haematoma and mobilization of duodenum and pancreas. PMID- 2967907 TI - Isotretinoin use in pregnancy. PMID- 2967908 TI - Vulval pruritus caused by by-mouth vitamin preparation. PMID- 2967909 TI - The endocrinology of extrapyramidal system disorders. AB - The study of endocrinologic changes in Parkinson's disease, Huntington's disease, and tardive dyskinesia may elucidate the pathophysiology of these disorders, especially the presence of hypothalamic lesions. There is probably a decrease in PRL concentrations in Parkinson's disease, and there may be an increase in TSH response to TRH stimulation. It is not clear if there is any change in GH concentrations in Parkinson's disease. There appears to be a robust increase in GH concentrations in Huntington's disease, and there may be a small increase in PRL as well. At present no endocrinologic abnormality has been well documented in tardive dyskinesia. PMID- 2967910 TI - An electrophoretic karyotype of Neurospora crassa. AB - A molecular karyotype of Neurospora crassa was obtained by using an alternating field gel electrophoresis system which employs contour-clamped homogeneous electric fields. The migration of all seven N. crassa chromosomal DNAs was defined, and five of the seven molecules were separated from one another. The estimated sizes of these molecules, based on their migration relative to Schizosaccharomyces pombe chromosomal DNA molecules, are 4 to 12.6 megabases. The seven linkage groups were correlated with specific chromosomal DNA bands by hybridizing transfers of contour-clamped homogeneous electric field gels with radioactive probes specific to each linkage group. The mobilities of minichromosomal DNAs generated from translocation strains were also examined. The methods used for preparation of chromosomal DNA molecules and the conditions for their separation should be applicable to other filamentous fungi. PMID- 2967911 TI - Opsin expression in the rat retina is developmentally regulated by transcriptional activation. AB - The gene for rhodopsin, the primary light sensor of the visual system, is specifically expressed in the rod photoreceptor cells of the retina. We show here that in the rat, opsin RNA first accumulates to detectable levels at postnatal day 2 (PN2) and that nascent transcripts can be detected at PN1; this is the time when peak numbers of photoreceptor cells are generated by the final division of their neuroepithelial precursors. Accumulated opsin RNA then increases to reach the adult level, 0.06% of total retinal RNA, at about PN10. The transcription rate of the opsin gene increases to a similar extent over the same time course between PN3 and adulthood, suggesting that transcriptional activation is responsible for the increase in opsin expression. We used the antibody RET-P1 to show that rhodopsin protein is also detectable at PN2 and that the number of cells expressing the protein increases with time in a central-to-peripheral gradient in the retina. This increase in the number of differentiating photoreceptors in the tissue appears to account for much of the increase in opsin gene transcription and RNA accumulation. In situ hybridization to opsin RNA shows that it is restricted to the photoreceptor layer from the time it can first be detected, at PN7. Later in development, when RET-P1 staining shifts to the photoreceptor outer segments, opsin RNA becomes localized to the inner segments, suggesting that the distributions of opsin protein and RNA are related. PMID- 2967914 TI - Characterization of the vasopressin receptor on WRK-1 cells. AB - Specific vasopressin binding to WRK-1 rat mammary tumor cells was assessed and compared with vasopressin-induced alterations in phosphatidylinositol metabolism. Scatchard analysis revealed the presence of two binding sites: a saturable, high affinity site with a dissociation constant of 1 X 10(-9) M and an n of 2700 sites per cell, and a nonsaturable, apparent lower affinity site. The higher affinity site appeared to have V1a specificity and to correlate with vasopressin's ability to stimulate phosphatidylinositol turnover in the cells. PMID- 2967912 TI - Interactions between cell growth-regulating domains in the products of the adenovirus E1A oncogene. AB - Among the various biological activities expressed by the products of the adenovirus E1A gene are the abilities to induce cellular DNA synthesis and proliferation in quiescent primary baby rat kidney cells. The functional sites for these activities lie principally within two regions of the E1A proteins: an N terminal region and a small second region of approximately 20 amino acids further downstream. To study the biological functions of the first domain, we constructed an in-frame deletion of amino acid positions 23 through 107 of the E1A products. This deletion did not impede the ability of the E1A products to transactivate the adenovirus early region 3 promoter in a transient-expression assay in HeLa cells. The ability to induce DNA synthesis in quiescent baby rat kidney cells was, however, lost in the absence of these sequences. Deletion of the small second region induced a form of S phase in which DNA synthesis occurred in the apparent absence of controls required for the cessation of DNA synthesis and progression through the remainder of the cell cycle. These cells did not appear to accumulate in or before G2, and many appeared to have a DNA content greater than that in G2. The functions of both domains are required for production of transformed foci in a ras cooperation assay. Focus formation occurred, however, even when the two domains were introduced on two separate plasmids. This complementation effect appeared to require expression of both of the mutant proteins and did not appear to result merely from recombination at the DNA level. PMID- 2967915 TI - Molecular structure of the chicken vitamin D-induced calbindin-D28K gene reveals eleven exons, six Ca2+-binding domains, and numerous promoter regulatory elements. AB - The seco-steroid hormone 1,25-dihydroxyvitamin D3 is known to induce the expression of a calcium binding protein termed calbindin-D28K in a variety of target tissues. In order to comprehend the mechanism of induction we have cloned and sequenced the chicken calbindin-D28K gene. The gene spans some 18.5 kilobases (kb) of chromosomal DNA from the putative Cap site to the polyadenylation site of the 2.8 kb mRNA. It is split into 11 coding exons by 10 intervening sequences. The promoter region of this gene is markedly G + C-rich (60-80%) extending from 225 to +400. Within this region we find 70 CpG dinucleotides, four G-C boxes, and numerous known promoter regulatory signals. These putative regulatory signals include a TATA box (ATAAATA) at -30 and a CAT box (CCAAT) at -326. Ten additional variant CAT boxes are found in the upstream promoter region (-218 to -770) of this gene. Furthermore we have identified a glucocorticoid-like responsive element at -410 (TCTACACACTGTTCC) and this element overlaps a metal responsive element (TGCACTC) and a variant CAT box (CCAAAT) and juxtaposes an enhancer-like core element (AAATGGT) on its 3'-side. In addition, the calbindin-D28K promoter is composed of a variety of simple repeated sequences, some of which are components of putative regulatory signals. All splice junctions were found to conform to the GT-AG rule. A consensus sequence of the 5'-splice junction reads AG/GTAAG-TTATA. A consensus sequence of the 3'-splice site consists of two elements: a pyrimidine track (mainly T) followed by ACAG/G-T. A two-dimensional model of calbindin-D28K was constructed which projects the existence of 6 alpha helix-loop-alpha-helix regions characteristic of calcium binding domains. The 3' end of the gene consists of a single large (2039 base pair) uninterrupted exon, an organizational feature common to other members of the calcium binding protein gene family which include calmodulin, parvalbumin, Spec I, myosin light chains, etc. Another feature common to the gene family is the presence of the repeated sequence ATTT or TTTA located in the 3'-untranslated exons. These simple repeat sequences could be involved in regulating mRNA degradation by serving as a ribonuclease recognition signal. PMID- 2967913 TI - Factors responsible for the higher transcriptional activity of extracts of adenovirus-infected cells fractionate with the TATA box transcription factor. AB - Extracts of adenovirus-infected HeLa cells have 5- to 10-fold-higher activity for transcription from the major late promoter in vitro than do extracts of mock infected or E1A mutant-infected cells (K. Leong and A. J. Berk, Proc. Natl. Acad. Sci. USA 83:5844-5848, 1986). In this study, we analyzed extracts from mock infected cells and from cells infected with an E1A mutant, pm975, which expresses principally the large E1A protein responsible for the stimulation of transcription. These extracts were fractionated by phosphocellulose chromatography, a procedure which separates factors required for transcription from this promoter (J. D. Dignam, B. S. Shastry, and R. G. Roeder, Methods Enzymol. 101:582-589, 1983), allowing the quantitative assay of individual factors (M. Samuels, A. Fire, and P. A. Sharp, J. Biol. Chem. 257:14419-14427, 1982). Fractions eluted with 0.04, 0.35, and 0.6 M KCl, which contained RNA polymerase II, the upstream factor MLTF, and three general polymerase II transcription factors, had similar activities when prepared from virus-infected or from mock-infected cells. The sequence-specific DNA-binding activity of MLTF was also similar in the virus-infected- and mock-infected-cell extracts. In contrast, the 1.0 M KCl fraction prepared from virus-infected cells consistently exhibited activity severalfold higher than that of the equivalent fraction prepared in parallel from mock-infected cells. E1A protein eluted principally (greater than 80%) in the 0.35 M KCl fraction. Results of others (M. Sawadogo and R. G. Roeder, Cell 43:165-175, 1985) have shown that the 1.0 M KCl fraction, containing 2 to 5% of the unfractionated protein extract, contains a factor which binds specifically to the major late promoter TATA box. These results, together with a recent genetic analysis of the E1B promoter which demonstrated that the TATA box was required for its efficient transcriptional activation (transactivation) by E1A (L. Wu, D. S. E. Rosser, M. Schmidt, and A. J. Berk, Nature (London) 326:512-515, 1987), are consistent with the model that E1A protein indirectly activates the TATA box transcription factor. Consistent with this model was the finding that mutants of the major late promoter containing only the TATA box and cap site region were transcribed at higher rates with extracts from virus-infected cells than with extracts from mock-infected cells. Other models consistent with the results are also discussed. PMID- 2967916 TI - Helper T-cell hybridomas as cellular probes for immunological investigation. PMID- 2967917 TI - Effect of captopril on progressive ventricular dilatation after anterior myocardial infarction. AB - We conducted a double-blind, placebo-controlled trial to determine whether ventricular dilatation continues during the late convalescent phase after myocardial infarction and whether therapy with captopril alters this process. Fifty-nine patients with a first anterior myocardial infarction and a radionuclide ejection fraction of 45 percent or less underwent cardiac catheterization 11 to 31 days after infarction, when they were not in overt congestive heart failure. They were randomly assigned to placebo or captopril and were followed for one year. A repeat catheterization was performed to evaluate interval changes in hemodynamic function and left ventricular volume. Thirty eight male patients were evaluated with maximal-exercise treadmill tests every three months. No differences were detected at base line in clinical, hemodynamic, or quantitative ventriculographic variables. During one year of follow-up, the end-diastolic volume of the left ventricle increased by a mean [+/- SEM] of 21 +/ 8 ml (P less than 0.02) in the placebo group, but by only 10 +/- 6 ml (P not significant) in the captopril group. The left ventricular filling pressure remained elevated with placebo but decreased (P less than 0.01) with captopril. In a subset of 36 patients who were at high risk for ventricular enlargement because they had persistent occlusion of the left anterior descending coronary artery, captopril prevented further ventricular dilatation (P less than 0.05). Patients given captopril also had increased exercise capacity (P less than 0.05). This preliminary study indicates that after anterior myocardial infarction, ventricular enlargement is progressive and that captopril may attenuate this process, reduce filling pressures, and improve exercise tolerance. PMID- 2967918 TI - A case report on a minor contamination of nivalenol in cereals harvested in Canada. AB - An investigation for the occurrence of nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN) in cereals (ten wheat, one rye and one corn) harvested in Canada have been carried out using a procedure, which is rapid and sensitive for Fusarium mycotoxins. NIV, DON and ZEN were detected in 4, 9 and 9 out of ten wheat samples, and their average levels in the positives were 23 ng/g, 1257 ng/g and 9 ng/g, respectively. One rye and one corn were also contaminated with a minor amount of NIV. This is the first evidence for the natural occurrence of NIV in cereals grown in Canada, though its level was far less than DON. PMID- 2967919 TI - [Medical examinations]. PMID- 2967921 TI - More about response to hepatitis B vaccine in hemodialysis patients. PMID- 2967920 TI - Atrial natriuretic peptide and arginine vasopressin in pregnancy and pregnancy induced hypertension. AB - Atrial natriuretic peptide (ANP) and arginine vasopressin concentrations were measured in 9 patients with pregnancy-induced hypertension. The results were compared to those found in 7 normal pregnant women matched for age, duration of pregnancy, and parity. Plasma ANP levels were significantly higher in the pregnancy-induced hypertension patients than in the control group. Plasma arginine vasopressin concentrations, however, were not significantly different in the two populations. The mechanism of the observed rise in ANP concentrations in the patients with pregnancy-induced hypertension is not known. However, it may be related to a rise in intra-atrial pressures secondary to hypertension, an increase in baroreceptor discharge as a result of hypertension, or, less likely, the ANP may be released from extracardiac sites. PMID- 2967922 TI - [Advances in the studies of the pathogenesis, treatment and prevention of genetically determined diseases of the extrapyramidal tracts]. PMID- 2967924 TI - Microsurgical versus standard lumbar discectomy. AB - Two hundred seventy consecutive patients operated upon by standard discectomy were compared to 270 patients treated with microlumbar discectomy. All patients had back and leg pain, as well as positive clinical findings and positive myelograms suggestive of disc herniation. The results demonstrated a 98% success rate in the microsurgical group as compared to a 95% success rate in the standard laminectomy group. Of microdiscectomy patients, 95% had an excellent result, as compared to 89% of the standard laminectomy group. Patients with pending compensation cases did only slightly poorer than those with no secondary gain regardless of what procedure was used. The postoperative hospital stay of the microsurgical patients averaged 3.7 days as compared to 7.1 for the standard technique. The time before return to work was significantly shorter in patients undergoing microdiscectomy. Microdiscectomy proved to be superior in both clinical results and cost effectiveness. PMID- 2967923 TI - [Ossification in the area of the spinal cord arachnoid as the cause of a chronic pain syndrome after spinal fracture]. AB - Calcifications and ossification in the spinal arachnoid are usually asymptomatic and are revealed accidentally during neurosurgical operations or autopsy. A case is described of ossification of the arachnoid developing after vertebral trauma and causing chronic pain. PMID- 2967925 TI - Adipose tissue in the filum terminale: a computed tomographic finding that may indicate tethering of the spinal cord. AB - Adipose tissue in the filum terminale is frequently associated with tethering of the spinal cord in patients with spina bifida occulta. We recently saw a patient with low back pain and no spina bifida occulta, in whom adipose tissue was noted in the area of the filum on an unenhanced computed tomographic (CT) scan. The patient had a tethered cord. This case suggested that, when CT scanning is done as the first imaging study in the evaluation of low back pain, fatty tissue in the area of the filum may be an indicator for tethering of the spinal cord. The present study was undertaken to determine the validity of using CT scan detectable filal fat in the identification of possible tethered spinal cords among a group of patients experiencing low back pain. The presence of fat in the fila of 12 patients with the radiologically and histologically tethered cord syndrome was evaluated and the fila of 47 autopsied patients whose clinical history showed no back pain were examined histologically. There were accumulations of adipose tissue in the fila of 11 of the 12 (91%) patients with the tethered cord syndrome and in the fila of 9 of the 47 patients (17%) in the autopsy series. Of the 9 autopsy patients with fat in their fila, however only 3 patients (6%) exhibited collections of adipose tissue in the CT detection range (2 mm).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2967926 TI - [Platelet aggregation in whole blood with the impedance method in subjects with non-complicated essential arterial hypertension]. AB - Arterial hypertension is considered a major risk factor in atherosclerosis in the pathogenesis of which platelet activity plays a fundamental role. However the data in the literature on platelet function in arterial hypertension do not always agree. The present study was conducted on whole blood, using the impedance metering technique to assess platelet aggregation induced by ADP (10 pg) and collagen (2 mg/ml) in 15 patients with uncomplicated essential hypertension and 25 healthy controls. Analysis of the data shows a statistically significant difference between the aggregation curves of the hypertensive and the healthy subjects with excessive platelet aggregation in those suffering from uncomplicated arterial hypertension. PMID- 2967927 TI - [Therapy of deep venous thrombosis with defibrotide. Pilot study]. PMID- 2967929 TI - A termination codon specifies selenocysteine in glutathione peroxidase. PMID- 2967928 TI - Contribution of cholinergic vasodilators on the increase in cerebral cortical blood flow responses to the intravenous administration of thyrotropin releasing hormone in anesthetized rats. AB - The effect of an intravenous administration of thyrotropin-releasing hormone (TRH) on the regional cerebral blood flow in the sensory cortex was studied in halothane-anesthetized adult Wistar rats. The regional cerebral blood flow was continuously monitored with the laser Doppler flowmetry. The cerebral blood flow increased dose-dependently following the administration of 3 x 10(2) and 3 x 10(3) microns/kg TRH. The systemic blood pressure also increased simultaneously. After maintaining the systemic blood pressure at a constant level via a pressure reservoir system, the TRH-induced increases in the cerebral blood flow continued to be observed. In atropinized animals, the blood pressure increased as high as that indicated in non-atropinized animals following TRH administration, but the responses of the cerebral blood flow in the latter were much attenuated. It was suggested that the cholinergic vasodilative system contributed to the TRH-induced increase in cerebral blood flow. PMID- 2967930 TI - Should nurses receive the hepatitis B vaccine? PMID- 2967931 TI - Hepatitis B vaccination. PMID- 2967933 TI - Low back pain. PMID- 2967932 TI - A combination of hepatitis B vaccine and immunoglobulin does not protect all infants born to hepatitis B e antigen positive mothers. AB - In 42 infants of hepatitis B e antigen positive carrier mothers a randomised study of a low dose (5 micrograms) hepatitis B vaccine regimen starting within 24 hours of birth with or without hepatitis B immunoglobulin (0.25 ml/kg) showed that in the vaccine group 72% were antibody positive at the age of one year and in the vaccine plus immunoglobulin group 81%. In the vaccine group four of 18 infants were not protected and in those infants receiving vaccine plus immunoglobulin three of 21 were HBsAg carriers by 12 months of age. Hepatitis B vaccine either with or without concomitant HBIg did not protect all babies born of HBeAg positive mothers, from developing the HBsAg carrier status. Thus for full protection of all infants a more vigorous immunisation programme will be necessary. PMID- 2967934 TI - Outpatient epidural steroid injections for low back pain and lumbosacral radiculopathy. AB - A survey was done of 250 patients with low back pain and sciatica, treated as outpatients in a pain relief practice, with epidural steroid injections. Repeated injections improved the success rate and provided a safe, cost effective means of treatment without the necessity of hospital admission. PMID- 2967935 TI - Duovent inhaler v Inhalets in patients with asthma and chronic bronchitis. PMID- 2967937 TI - Management of backache. PMID- 2967936 TI - A dose response study comparing Duovent vs salbutamol. AB - The dose response profiles of Duovent (fenoterol 0.1 mg/ipratropium 0.04 mg per puff) and salbutamol (0.1 mg per puff) were determined in a double-blind, controlled study. Twenty-one patients with asthma and nine patients with chronic bronchitis received 1, 2, 4, and 6 puffs of either Duovent or salbutamol on separate days. FEV1, pulse and tremor were recorded at baseline and up to 360 minutes after inhalation of test medication. Duovent and salbutamol produced an effective bronchodilator response at all dose levels, however, at 2, 4 and 6 puffs, the improvement with Duovent was significantly greater than with salbutamol alone in both asthmatic and chronic bronchitic patients. Two puffs of Duovent produced a significantly greater bronchodilation than all doses of salbutamol. There was no difference in the bronchodilator response between 1-6 puffs of salbutamol indicating maximal dose was achieved with one puff. There was an incremental improvement in FEV1 with 1, 2, and 4 puffs of Duovent but no difference between 4 and 6 puffs. There was no difference in pulse rate or tremor between salbutamol or Duovent at any dose level. We conclude that Duovent produces a greater and more prolonged bronchodilator response than salbutamol in both asthmatic and chronic bronchitic patients. PMID- 2967938 TI - Social factors in chronic pain rehabilitation. PMID- 2967939 TI - Atrial natriuretic factor and the heart in health and disease. PMID- 2967941 TI - AIDS update. PMID- 2967940 TI - Antibody responses to recombinant, yeast-derived hepatitis B vaccine in teenage New Zealand children. AB - Three groups of healthy teenage New Zealand children were given 2.5 micrograms, 5 micrograms and 10 micrograms, which is the currently recommended dose, of Merck Sharp and Dohme recombinant yeast-derived hepatitis B vaccine at time 0, 1 and 6 months and tested for antibody responses to vaccine and for other hepatitis B virus markers. Seroconversion rates exceeded 98% in all three groups. Geometric mean titres (GMT) of the anti-HBs increased with higher doses. There was no significant differences in GMT between the sexes. Under the conditions of this study, 2.5 micrograms doses of this vaccine induced an excellent antibody response in children 12-14 years of age. PMID- 2967942 TI - Quality dental education as a fundamental aspect of its responsibility to the community. PMID- 2967943 TI - Serum CA 125 measurements in the diagnosis of endometriosis recurrence. AB - Seventy-one women underwent follow-up laparoscopy for persistent infertility six to 24 months after conservative treatment of endometriosis. The disease was staged according to the revised American Fertility Society classification of 1985. The serum CA 125 concentration was also measured in each patient to evaluate its efficacy in the diagnosis of endometriosis recurrence. The patients with stages I and II endometriosis had serum CA 125 levels not significantly higher than in the patients with negative findings, whereas those with stages III and IV endometriosis presented significantly higher levels (P less than .005 and P less than .001, respectively) than the disease-free women. The sensitivity of serum CA 125 measurements in the diagnosis of endometriosis recurrence was 14.8%, the specificity was 100%, and the predictive values of normal (less than 35 U/mL) and elevated levels were 27 and 100%, respectively. In some cases, serum CA 125 measurements may be used instead of follow-up laparoscopy, or to indicate when laparoscopy should be performed or postponed. PMID- 2967944 TI - Who buys the groceries if you're disabled? PMID- 2967946 TI - When back pain becomes disabling: a regional analysis. AB - Back pain is a common condition and in most cases is not disabling. We have investigated disabling back pain that leads to health care utilization, time lost from work, and high costs. Disabling back pain remains of obscure origin because the focus in studying it has been too narrow. Our indicator of disability is the industrial insurance claim rate for back sprain by county (N = 39) in the State of Washington. After controlling for the size of the labor force and the proportion of workers in occupations that are particularly at risk of back sprain, we determined the effect of 3 socioeconomic factors on the claim rate: the unemployment rate, percentage receiving food stamps, and per capita income. For 2 of the 3 years studied, socioeconomic factors accounted for about one-third of the variance in the claim rate. Even though claimants of industrial insurance are employed, the unemployment rate was significantly related to the claim rate in the 3 years studied. Our interpretation is that disability is a symptom of distress. Where there is a rise in job insecurity and an attendant rise in economic insecurity, there is a greater likelihood that back pain will become disabling. PMID- 2967945 TI - Prediction of pain behavior and functional status of rheumatoid arthritis patients using medical status and psychological variables. AB - This study examined the extent to which the psychological variables of depression, anxiety, and helplessness predicted the pain behavior and functional status of 64 rheumatoid arthritis (RA) patients beyond what could be predicted on the basis of demographic and medical status variables. Pain behavior was evaluated using a standardized observation method, and functional status was assessed using a modified Health Assessment Questionnaire (MHAQ) and rheumatologists' ratings. Regression analyses revealed that a modified rheumatoid activity index and/or disease duration were significant predictors of levels of guarding, rigidity, and total pain behavior. The psychological variables examined did not predict independently RA pain behavior. The rheumatoid activity index explained a significant proportion of the variance in functional status ratings and MHAQ daily function scores. Age, disease duration and depression also were independent predictors of functional status ratings. Thus, depression had a significant relationship with physician ratings of functional status but not with patient self-reports of disability. Psychological factors not examined in this study that might influence RA pain behavior and self-reports of functional status are discussed. PMID- 2967947 TI - Alterations in cerebrospinal fluid concentrations of somatostatin-like immunoreactivity in chronic pain patients. AB - Concentrations of somatostatin-like immunoreactivity in cerebrospinal fluid were significantly reduced in chronic pain patients compared to control patients without chronic pain. This difference was not influenced by demographic or clinical characteristics. Somatostatin has been shown to be a neurotransmitter in animal nociception; pharmacologic doses of this substance have moderated human pain. Our findings provide evidence that somatostatin may be involved in the pathogenesis of the chronic pain state. PMID- 2967948 TI - Bacterial infections in human immunodeficiency virus-infected children. AB - A retrospective review of 71 children infected with human immunodeficiency virus cared for over a 3.5-year period revealed that 44 of 71 (63%) required a bacterial culture and 27 of 71 (37%) had bacteriologically documented infection. There were 125 episodes in 27 patients. Pneumonia (24 of 125 (19%)), upper respiratory tract syndromes (23 of 125 (19%)), urinary tract infection (24 of 125 (19%)) and wound infection (12 of 125 (10%)) were the most common syndromes identified. Bacteremic infections occurred in 35 of 125 (28%), and in 17 of 125 (14%) no other primary source could be identified. Pneumococci (11 of 35 (31%)) and Salmonella (4 of 35 (11%)) were the most common blood isolates; however, a wide spectrum of Gram-positive and Gram-negative pathogens were recovered. Bacterial pneumonia directly contributed to the death of 4 patients, in whom pneumonia caused by Pneumocystis carinii (2), cytomegalovirus (1) or varicella zoster virus (1) also coexisted, respectively. Absolute T4 counts less than 400 and depressed lymphocyte-proliferative responses to diphtheria and tetanus toxoids, Candida antigen and pokeweed mitogen correlated with the occurrence of bacterial infection in human immunodeficiency virus-infected children. Although bacterial infections are a frequent cause of morbidity in human immunodeficiency virus-infected children, they are usually treatable. PMID- 2967949 TI - [Experience in organizing the dispensary observation in the pediatric ward for children with chronic pathology]. PMID- 2967950 TI - [Work experience of the Astrakhan oral rehydration center]. PMID- 2967951 TI - [Diagnostic error in the Bland-White-Garland syndrome and thymomegaly in a nursing infant]. PMID- 2967952 TI - [Pituitary gonadotropic adenoma]. PMID- 2967953 TI - [Postpartum myocardiopathy. Clinical and echographic evaluations of the response to treatment. 30 cases observed in the Sudan-sahelian area]. AB - Thirty cases of postpartum cardiomyopathy observed in a savannah-sahelian region of Africa (Niamey, Republic of Niger) are reported. The dilated cardiomyopathy was diagnosed by comparing clinical signs with electrocardiographic, radioscopic and echocardiographic findings. A simplified therapeutic regimen was used in all cases: rest during 2 months, sodium restriction, diuretic and digoxin therapy (1 tablet of each drug every other day). Patients were followed up for a mean period of 13 months. The course of the disease was evaluated at regular intervals by echocardiography which provided accurate information on left ventricular function. At the time of diagnosis all patients had congestive heart failure. Complete remission was defined as the absence of all signs and a normal social life. Twelve patients answered this definition with a mean follow-up of 13 months. Nine patients (mean follow-up 24 months) were in incomplete remission; a large left ventricular aneurysm had developed in 2 of them. In 6 other patients the follow-up was too short (1 to 8 months) for a prognosis to be formulated. In postpartum cardiomyopathy 3 factors are indicative of a poor prognosis: 1) absence of free interval between delivery and first signs of heart failure; 2) history of cardiomyopathy after previous deliveries; 3) poor response to treatment with persistent cardiomegaly. On the other hand, the severity of the initial cardiomegaly and impairment of ventricular contractility, and the presence of permanent hypertension or pericardial effusion have no significant influence on the response to treatment. Irreversible contraception is justified in great multiparous women. In primiparous and oligoparous women a temporary contraception is necessary, a new pregnancy being conceivable only after complete remission and under close supervision. PMID- 2967954 TI - [Retrospective bacteriological study of mycobacterial infections in patients with acquired immunodeficiency syndrome]. AB - The main species of mycobacteria isolated in 62 of the 316 acquired immunodeficiency syndrome patients admitted to the Claude Bernard Hospital, Paris, between January, 1983 and October, 1986 were studied retrospectively according to their site of isolation and their pathogenic role. Mycobacterium tuberculosis was isolated in 19 cases (from pulmonary specimens in 17 cases); this species was present in 59 percent of our African patients as against 20 percent of our European patients. M. avium intracellulare was isolated in 33 cases (17 from blood, 12 from the lung and 11 from the gastrointestinal tract) and was found in 55 p. 100 of our European patients. Other species that were isolated less frequently were M. xenopi (5 cases), M. kansasii (3 cases), M. aurum, M. chelonae, M. fortuitum, M. gordonae, M. simiae and M. terrae (1 case each). Post mortem specimens obtained from 110 acquired immunodeficiency syndrome patients were cultivated during the same period. In 20 patients, at least one specimen was positive for a mycobacterium: M. tuberculosis in 2 cases, M. avium intracellulare in 18 cases. Twenty-nine of the 33 patients in whom M. avium intracellulare was isolated were considered a posteriori as being infected by this organism. The therapeutic approach varies according to the species involved. No treatment seems to be truly effective against M. avium intracellulare. Pending the results of cultures, no direct bacteriological examination can provide information on the mycobacterial species concerned; however, a conventional antituberculosis treatment may be instituted, particularly in patients from Africa or Haiti. PMID- 2967955 TI - [Listeria rhombencephalitis caused by Listeria monocytogenes with a cerebrospinal fluid initially normal]. AB - A case of rhombencephalitis due to Listeria monocytogenes in a non immunocompromised patient, with initially normal cerebrospinal fluid, was marked by potentially fatal neurovegetative disorders and severe neurological sequelae partly due to delay in diagnosis and treatment. The possibility of Listeria infection should be considered in patients with fever and cranial nerves deficits, in order to initiate an appropriate antibiotic therapy and to keep the subject under close monitoring in an intensive care unit. PMID- 2967956 TI - [Role of toxoplasma and viruses in polydermatomyositis]. AB - The causes of polymyositis are imperfectly known. The role of Toxoplasma, the myotropism of which is significant in acute myositis, may be suspected on the ground of secondary serological data, but it has not yet been confirmed. On the other hand, the role of coxsackie virus, which until recently had been established only on morphological or serological data, has now been ascertained by molecular biology techniques. The best example of experimental viral polymyositis is the disease induced by inoculation of retrovirus to the monkey. This has led to the identification in man of polymyositis revealing an acquired immunodeficiency syndrome. PMID- 2967957 TI - [Renal transplantation in thrombosis of the inferior vena cava. Implantation of the graft vein in the portal vein]. AB - An 8-year old girl with chronic renal failure underwent allogeneic renal transplantation with implantation of the renal vein of the graft on the portal vein, as the inferior vena cava was obstructed by thrombosis. The possible technical obstacles to renal transplantation are reviewed on that occasion, and solutions are suggested. In the presence of thrombosis of the inferior vena cava it seems necessary to determine its level and to look for a patent venous segment all the way up to the diaphragm. If no such segment is found, then the renal vein can be implanted on the portal vein. PMID- 2967958 TI - [Interaction of bilirubin on the oxyhemoglobin saturation measured by photometry]. PMID- 2967959 TI - [Epileptic seizures of the supplementary motor area in hyperglycemia without ketosis]. PMID- 2967960 TI - [Pulmonary alveolar echinococcosis]. PMID- 2967961 TI - [Syndrome of acute respiratory distress with a fatal development in a treatment with D-penicillamine]. PMID- 2967962 TI - [Torulopsis glabrata endocarditis after cardiac surgery for heart valve replacement]. PMID- 2967963 TI - [A rare cause of bilateral serofibrinous pleurisy: ankylosing spondylitis]. PMID- 2967964 TI - [Bilateral apical pulmonary edema: an unusual manifestation of acute mitral insufficiency]. PMID- 2967965 TI - [Infected pseudo-myxoma of the left atrium: large endocardial vegetation of the mitral valve with a favourable development with antibiotic therapy]. PMID- 2967966 TI - [Loss of teeth in scleroderma]. PMID- 2967967 TI - [Outcome of neurosyphilis in the development of acquired immunodeficiency syndrome]. PMID- 2967968 TI - [Persistent fever after appendectomy, disclosing a Yersinia infection in a patient with hemochromatosis]. PMID- 2967969 TI - Putative metabolites derived from dietary combinations of calcium glucarate and N (4-hydroxyphenyl)retinamide act synergistically to inhibit the induction of rat mammary tumors by 7,12-dimethylbenz[a]anthracene. AB - Calcium glucarate and N-(4-hydroxyphenyl)retinamide were evaluated individually and in combination in the diet as preventative chemical agents, by using the induction of rat mammary tumors by 7,12-dimethylbenz[a]anthracene as the test system. When tested separately over 18 weeks, optimal doses of calcium glucarate (128 mmol/kg of diet) or N-(4-hydroxyphenyl)retinamide (1.5 mmol/kg of diet) administered daily inhibited tumor incidence by 50% or 57% and tumor multiplicity by 50% or 65%, respectively. Suboptimal doses of calcium glucarate (32 mmol/kg) and of N-(4-hydroxyphenyl)retinamide (0.75 mmol/kg) inhibited tumor incidence by 15% and 5% but had no inhibitory effect on tumor multiplicity. In contrast, the combination of calcium glucarate (32 mmol/kg) and N-(4-hydroxyphenyl)retinamide (0.75 mmol/kg) inhibited tumor incidence and tumor multiplicity by 50%. Similar synergism was observed with the combination of calcium glucarate (64 mmol/kg) and N-(4-hydroxyphenyl)retinamide (0.75 mmol/kg), the inhibition being 55-60%. HPLC analysis of the bile of female rats injected intraperitoneally with a single dose of the retinamide [60 mg/kg (body weight)] showed that the excretion of the retinamide and its glucuronide were markedly suppressed by pretreatment with an oral dose of calcium glucarate [4.5 mmol/kg (body weight)]. PMID- 2967970 TI - Trans-activation of the human immunodeficiency virus long terminal repeat sequences, expressed in an adenovirus vector, by the adenovirus E1A 13S protein. AB - The human immunodeficiency virus 1 (HIV-1) long terminal repeat (LTR) sequences were inserted into adenovirus in place of the E1 region. The HIV-1 LTR contained in this recombinant adenovirus responds to trans-activation by tatIII in a HeLa cell line constitutively expressing that HIV-1 gene product. In addition, the HIV 1 LTR is activated by the adenovirus E1A 13S, but not 12S or 9S, gene product when it is supplied in trans by a coinfecting wild-type adenovirus. The Rous sarcoma virus LTR, in a similar recombinant adenovirus, is insensitive to tatIII but is also trans-activated by the E1A 13S protein. The action of the 13S E1A and tatIII proteins are additive for the HIV-1 LTR in the context of adenovirus and they appear to act at the transcriptional level. As in HeLa cells, the adenovirus borne HIV-1 LTR is inactive in the absence of a trans-activator in H9 and Jurkat cells, two human leukemic T-cell lines. This suggests that recombinant adenoviruses have diagnostic potential for the detection of trans-activators of the HIV-1 LTR that are present in circulating human lymphocytes. PMID- 2967972 TI - Effect of carcinogenic nitroso compounds on rat kidney proximal tubule NAD levels. AB - An exposure of proximal tubules to alkylating nitrosamines is not necessary to deplete their NAD levels. Further work is needed to test if the poly(ADP-ribose) transferase induction theory accounts for the observed depletion of intracellular NAD. Also the viability of proximal tubule cells in the conditions of the study must be determined. Phosphate uptake by BBM vesicles prepared from chilled proximal tubules compared to those from proximal tubules incubated at 37 degrees C is a useful model for examining the role of intracellular NAD in renal Na+ gradient-dependent Pi transport. More experiments are required to assess the significance of changes in Pi-uptake by BBM vesicles induced by depletion of proximal tubule NAD content. PMID- 2967971 TI - Kinetics of gamma interferon binding and induction of major histocompatibility complex class II mRNA in Leishmania-infected macrophages. AB - Cells of the monocyte-macrophage series must carry out discrete accessory-cell functions during the process of antigen-specific T-cell activation. One of these functions is the cell-surface expression of major histocompatibility complex (MHC) class II gene products, which are involved in the presentation of foreign antigen to T cells. Previously, we reported that murine peritoneal macrophages infected with the obligate intracellular protozoan Leishmania donovani had suppressed responses to gamma interferon (IFN-gamma) for the induction of MHC class II antigen expression. To determine the molecular basis for this suppression, we examined in the present series of experiments the interaction of this organism with cells of the murine macrophage tumor cell line P388D1. When infected with Leishmania, these cells were also markedly unresponsive to IFN gamma for the induction of MHC class II antigen expression. This finding was not the result of a defect at the level of the IFN-gamma receptor. Thus, when 125I labeled IFN-gamma was used, infected macrophages were found to express normal numbers of high-affinity IFN-gamma receptors, and ligand-receptor binding resulted in rapid internalization of labeled IFN-gamma. Despite normal ligand receptor interactions, the induction in infected cells of mRNA encoding MHC (H-2) class II I-A alpha and beta chains in response to IFN-gamma was markedly suppressed. However, infected cells had normal levels of mRNA encoding the cytoskeletal protein actin. These findings indicate that Leishmania interferes with IFN-gamma induction of macrophage MHC class II antigen expression by down regulating lymphokine induction of MHC class II mRNA. Suppression of class II expression by this intracellular parasite may prevent subsequent T-cell recognition of infected macrophages and thus favor parasite survival. PMID- 2967973 TI - Effect of an atrial natriuretic peptide, anaritide, on renal function in dogs on high- and low-sodium diets. AB - These studies examined the effects in dogs of dietary Na intake on plasma "ANF like" immunoreactivity (PAI), and on the renal response to anaritide, a synthetic natriuretic peptide. There were no significant differences between high-Na (120 mEq/day) and low-Na (3 mEq/day) dogs in endogenous PAI, nor was PAI altered by oral or i.v. volume expansion with 0.9% saline. Renal arterial infusion of anaritide did not alter arterial pressure, renal blood flow or the frational excretion of lithium, but increased the fractional excretion of Na to similar degrees in both groups. The peptide increased GFR only in the low-Na group. Extremes of sodium balance do not seem to alter the renal response to anaritide, which apparently does not inhibit sodium reabsorption in the proximal tubule. PMID- 2967974 TI - Response of toad urinary bladder to vasopressin analogs possessing V1 or V2 specificity. AB - The urinary bladder of the toad Bufo marinus has been used to examine the effect on sodium transport, measured by short-circuit current, of natural antidiuretic hormones and several synthetic peptide analogs. In mammals, these synthetic analogs show specificity for different receptors, designated V1 and V2 receptors, whose biological responses are mediated by phosphatidyl inositol breakdown products or adenylate cyclase activity, respectively. All analogs stimulated SCC, with relative potencies AVT greater than AVP greater than Phe2 OVT (V1 agonist) much greater than d(CH2)5Tyr(Me)AVP (V1 antagonist) = d(CH2)5[D-Ile2,Abu4]AVP (V2 antagonist). The V1 and V2 antagonists inhibited the SCC response to AVT and Phe2OVT, with similar inhibitory potencies. We conclude that the stimulation of sodium transport by antidiuretic hormones involves one hormone receptor which does not show the selectivity of mammalian antidiuretic hormone receptors, and may represent a more primitive type of receptor. PMID- 2967975 TI - Mechanisms of increased shortening of sensitized airway smooth muscle. PMID- 2967976 TI - Anti-cholinergic agents and their use in asthma. PMID- 2967977 TI - Acute effects of fenoterol (Berotec) and ipratropium bromide (Atrovent) alone and in combination on bronchial hyperresponsiveness in asthmatic subjects. PMID- 2967978 TI - Fixed dose combination therapy in the treatment of asthma--the case against it. PMID- 2967979 TI - Biochemical mechanisms of oxfenicine cardiotoxicity. AB - Oxfenicine (S-4-OH-phenyl-glycine) was proposed as a compound which would stimulate carbohydrate utilization in the heart and thus reduce oxygen requirement, especially in ischemic heart disease. Oral administration to rats for several weeks gave rise to an increase in heart, liver and kidney weights. The drug damaged mitochondrial metabolism, reducing oxygen consumption and uncoupling oxidative phosphorylation in all three organs. In heart mitochondria creatine phosphate kinase was inhibited and the creatine content of the mitochondria increased. Myocyte membrane functions (Ca uptake as well as Na/K-, Mg- and Ca-ATPases) were inhibited. In all three organs lipids (phospholipids and triglycerides) as well as free fatty acids showed a transient accumulation. PMID- 2967980 TI - Photoinduced degradation and modification of Photofrin II in cells in vitro. PMID- 2967981 TI - The effect of photodynamic therapy alone and in combination with misonidazole or X-rays for management of a retinoblastoma-like tumour. PMID- 2967982 TI - Intracellular fluorescence of photosensitizing porphyrins at different concentrations of mitochondria. PMID- 2967983 TI - Down syndrome. PMID- 2967984 TI - Lower-extremity deep venous thrombosis: further experience with and refinements of US assessment. AB - During a 1-year period, 537 compression ultrasound (CU) examinations were performed in 506 extremities of 485 patients for evaluation of possible deep venous thrombosis (DVT). Decisions regarding therapy were based on the result of this test in both outpatients and inpatients. Doppler ultrasound was also used during the last 7 months of the study, but this procedure did not alter the results obtained with CU alone. Incidental findings (eg, Baker cysts, aneurysms, and lymph nodes) were noted in 42 patients and were thought to be nonthrombotic causes of leg swelling or pain that could not be detected with venography. The reliability and clinical acceptance of CU as a means of diagnosing lower extremity DVT resulted in a 50% yearly reduction in venography, with a concomitant 130% increase in the number of cases of DVT diagnosed. PMID- 2967985 TI - Percutaneous laser thermal angioplasty: initial results and 1-year follow-up in 129 femoropopliteal lesions. AB - Percutaneous peripheral laser thermal angioplasty with a laser-heated metallic capped fiber was used as an adjunct to conventional balloon angioplasty. Initial angiographic and clinical success was achieved in 99 of 129 (77%) femoropopliteal stenoses and occlusions (21 of 22 [95%] stenoses, 17 of 17 [100%] short [1-3-cm] occlusions, 26 of 37 [70%] medium-length [4-7-cm] occlusions, and 35 of 53 [66%] long [greater than 7 cm] occlusions). There was a 4% frequency of vessel perforation without clinical sequelae and no necessity for emergency bypass surgery. The 1-year cumulative clinical patency was 77% for the 99 lesions with an initial clinical success. In the 21 stenoses and 17 short occlusions, the cumulative clinical patency rates were 95% and 93%, respectively. In the longer occlusions (4-7 cm and greater than 7 cm), the clinical patency rates were 76% and 58%, respectively. The initial angiographic and clinical success, as well as the 1-year cumulative clinical patency, for stenoses and short occlusions after laser-assisted balloon angioplasty may be greater than after conventional balloon angioplasty alone. PMID- 2967986 TI - Occlusion during iliac angioplasty: a salvageable complication. AB - During transluminal dilation of the iliac artery, occlusion resulting from dissection occurred in four patients. In all four, the deteriorating clinical findings prompted surgical intervention. In three patients, Fogarty balloon catheters easily passed the occluded segments and specimens much the same as surgical endarterectomy specimens were retrieved. A clamp was used to retrieve the dissected portion of the vessel wall in the fourth patient. Three of four vessels have remained patent for 18 months, 18 months, and 6 months, respectively. One patient underwent bypass surgery 4 months after the occlusion episode for recurrent stenosis in a segment of vessel above the occluded segment, which had also been dilated during the same procedure. It is therefore possible in some cases to salvage vessels occluded during angioplasty, making it unnecessary to resort to aortofemoral or other type of bypass. PMID- 2967987 TI - Common iliac artery occlusion: treatment with pull-through angioplasty. AB - A technique was developed to traverse an occlusion of the common iliac artery when approach from an ipsilateral puncture has been unsuccessful. The technique involves an antegrade approach to the occlusion and allowed successful passage in seven cases. At 2-year follow-up, only one of the seven occlusions had recurred. PMID- 2967988 TI - Wire-loop technique for angioplasty of total iliac artery occlusions. AB - To recanalize occlusions of the iliac artery from an ipsilateral approach, the authors developed a technique in which the guide wire is manipulated through puncture sites in both sides of the groin. This technique was used in four occlusions (three in the common iliac artery and one in the external iliac artery); all occlusions were successfully dilated with balloon catheters. In two cases in which dilation was performed from the contralateral side, the contralateral iliac artery became occluded after the procedure. These occlusions were successfully dilated. PMID- 2967989 TI - [Recalibration of the lumbar canal, an alternative to laminectomy in the treatment of lumbar canal stenosis]. AB - A technique is described for widening the lumbar vertebral canal in the treatment of lumbar stenosis. This method, which preserves part of the posterior arches, has the aim of avoiding the instability which accompanies extensive laminectomies of the whole lumbar spine. The preservation of the posterior arches allows a ligamentoplasty to be made with dacron in stable spines and a combined fusion with internal fixation applied to the posterior arches to be made in cases of spondylolisthesis. The results of a prospective series of 32 patients have confirmed the effectiveness of the method both on the stenosis and on the post operative spinal stability. PMID- 2967990 TI - [Long-term results of interbody anterior arthrodesis in the treatment of common low back pain. Importance of preoperative discography findings]. AB - The results of 50 anterior interbody arthrodeses performed for incapacitating chronic low back pain with disc degeneration were assessed with a mean follow-up of seven years and a rate of examination of 74.5 per cent. Systematic pre operative discography was made of the levels above and below the fused region. The overall functional results were assessed using a new scale of evaluation which showed that only half of the patients were definitely improved at the latest follow-up examination. The outcome in relation to pain radiating to the lower limbs seemed to be the same as in low back pain and there was therefore no need for any posterior procedure. The factors that improved the success rate were an effective fusion, which was obtained in 80 per cent of cases at the first attempt and a healthy disc at the new effective lumbosacral level. The factors that diminished the success rate were mainly those relating to social and working activity, mainly the effects of accidents at work. Even though the results were less than perfect, interbody fusion, which the authors continue to practise, cannot be excluded from the therapeutic methods used to manage low back pain. PMID- 2967991 TI - Immunoassay detection of drugs in racing horses. IV. Detection of fentanyl and its congeners in equine blood and urine by a one step ELISA assay. AB - We have developed and evaluated a one step enzyme-linked immunosorbent assay (ELISA) test for fentanyl as part of a panel of pre- and post-race tests for narcotic analgesics in racing horses. This ELISA test detects fentanyl with an I 50 of about 100 pg/ml. The test is economical in that it can be read with an inexpensive spectrophotometer, or even by eye. The test is rapid, and ten samples, a normal pre-race complement, can be analyzed in about twenty minutes. The test readily detects the presence of fentanyl or its metabolites in equine blood and urine from two and twenty-four hours respectively after administration of sub-therapeutic doses. The two antibodies evaluated also cross-react with the methylated analogs of fentanyl, sufetanil and carfentanil and the test detected these drugs shortly after their administration to horses. When introduced into routine screening, this test, in combination with another immunoassay test previously described, yielded 10 sufentanil positives. As such this test is capable of both improving the quality and reducing the cost of pre-race and post race testing for fentanyl and a number of its congeners in racing horses. PMID- 2967992 TI - [Role of arthrosis in the diagnosis of backache]. PMID- 2967993 TI - [Anatomical review of the abdominal wall and physiopathology of hernias]. PMID- 2967994 TI - Back pain in middle age; occupational workload and psychologic factors: an epidemiologic survey. AB - The prevalence of back pain in a sample of 575 fifty-five-year-old residents of Malmo, Sweden, was studied and predisposing factors were analyzed. The point prevalence of back pain was 29%. Individuals with back pain had been less successful in a childhood intelligence test, had a shorter education and worked at physically more strenuous jobs. They were also less satisfied with their working conditions. PMID- 2967996 TI - Nerve root infiltration in the diagnosis of radicular pain. AB - Clinical and standard radiographic evaluation of patients with lumbosacral radicular symptoms may, on occasion, fail to delineate a cause. This study retrospectively reviews 62 patients who had undergone nerve root infiltration (NRI) and assesses the accuracy and indications for this diagnostic study. Surgical exploration of patients with a Group 1 response (typical pain reproduced by needle placement and then relieved by NRI) confirmed local root pathology in all. Exclusive of patients with arachnoiditis, a Group 1 response showed 85% accuracy in identifying a single symptomatic root. A Group 2 response (typical pain reproduced by needle placement but not relieved by local anesthesia) indicated multiple root involvement. Patients with a Group 3 or Group 4 response (typical pain not reproduced by needle insertion, with or without relief of pain by local anesthesia) were seldom relieved of radicular pain. NRI was most useful in investigation of patients with radicular symptoms in whom other investigations were 1) normal, 2) showed multiple level involvement, or 3) were difficult to interpret because of previous surgery. PMID- 2967995 TI - Painful lumbar end-plate disruptions: a significant discographic finding. AB - Of the 692 discs injected during lumbar discograms, end-plate disruptions with leakage of contrast material into the vertebral bodies were noted in 14 discs. Although gentle pressure was applied during the injections, severe fully concordant pain was reproduced in four (28.3%) discs, moderately severe and fully concordant pain in nine (64.3%) discs, and mild discordant pain in one (7.4%) disc. This is compared to 11.2% of the remaining 678 discs without end-plate disruption that reproduced severe concordant pain, 31.1% with moderately severe concordant pain, 17.1% with mild pain, and 40.6% without any pain reproduction. The difference between pain frequency in discs with end-plate disruption and those without is statistically significant (P less than .001). This suggests that end-plate disruptions may be related to painful segments. PMID- 2967998 TI - [Technical aids]. PMID- 2967997 TI - Surgical management of isthmic spondylolisthesis. AB - In order to determine the factors that produce a successful outcome in the surgical management of spondylolisthesis, 44 consecutive patients who underwent surgical treatment were reviewed. Preoperative evaluation was directed towards determining the presence or absence of nerve root compression. The patients without nerve root compression typically underwent fusion alone, while those with evidence of nerve root compression had decompression and fusion. After an average 36-month follow-up, results were good in 78%, fair in 18%, and poor in 4%. There were five complications, all of which were successfully managed. Two patients required reoperation for pseudoarthrosis. An eventual 100% fusion rate was achieved. Results in the patients with Grade III or Grade IV olisthesis were nearly equivalent to the less severe cases. Nine patients received allograft, with a fusion rate equal to the 32 receiving autogenous bone. This study suggests that in patients who have isthmic spondylolisthesis with mechanical lumbar symptoms only, or in patients with low-back pain with radicular extremity pain caused by nerve root irritation, fusion alone is indicated. In patients with documented radiculopathy resulting from nerve root compression, decompression in addition to fusion is indicated. PMID- 2967999 TI - [Reflections on the impact of malformations of the extremities]. PMID- 2968000 TI - Simultaneous measurement of hepatic arterial and portal venous flows by transit time ultrasonic volume flowmetry. AB - Simultaneous measurement of the hepatic artery and the portal vein was performed successfully upon 15 anesthetized patients during abdominal operations with the use of transit time ultrasonic volume flowmeter. The hepatic arterial flow, portal venous flow and total hepatic flow were 267.3 +/- 21.2, 746.4 +/- 41.3 and 1,010.7 +/- 52.7 milliliters per minute, respectively. The ratio of hepatic arterial flow to portal venous flow was 0.36 +/- 0.03. Temporary occlusion of the portal vein resulted in a significant increase in hepatic arterial flow (23.6 +/- 4.3 per cent, p less than 0.01), whereas temporary occlusion of the hepatic artery did not alter portal venous flow significantly. Occlusion of the common hepatic artery induced a significant decrease in hepatic arterial flow (p less than 0.05), but did not alter portal venous flow significantly. The results of this approach had useful clinical meanings in one patient in whom the Appleby's operation was performed in which sufficient hepatic arterial flow was needed after the common hepatic artery was ligated. Data are presented for hepatic circulation measured by transit time ultrasonic volume flowmeter, which is a good device to use to assess portal venous flow as well as hepatic arterial flow with reproducibility and stability. PMID- 2968001 TI - Removal of neointimal hyperplasia in Dacron grafts by an oscillating ring stripper. PMID- 2968002 TI - The hypotensive mechanism of percutaneous transluminal dilatation (PTD) in renovascular hypertension due to bilateral renal artery stenosis. AB - The hypotensive mechanism of percutaneous transluminal dilatation (PTD) in renovascular hypertension due to bilateral renal artery was investigated in two patients. Blood pressure was monitored continuously before, during and after PTD by use of a new automated blood pressure monitoring device based on finger volume oscillometry. Plasma renin activity was measured repeatedly before, during and after PTD. A hypotensive effect appeared immediately after PTD and blood pressure remained low in the following observation period without any hypotensive medication. In these cases, the hypotension was accompanied by a transient decrease in heart rate immediately after PTD. The hypotensive response to PTD was not parallel to the basal plasma renin activity, suggesting that the renin angiotensin system is not necessarily involved in the maintenance of the hypertension before PTD. The autonomic nervous system seemed to play a certain role. Since the hypotension was accompanied by a transient decrease in heart rate immediately after PTD, the hypotension may be induced either by a decrease in sympathetic tone or by an increase in vagal tone at least just after PTD. It is hypothesized that these changes in autonomic nervous activity are mediated centrally through the renal afferent mechanism in response to rapid changes in renal hemodynamics induced by PTD. PMID- 2968003 TI - Induction of the CD4-8+ suppressor phenotype in CD4+8+ human thymocytes by phorbol myristate acetate. AB - Human thymocytes were separated according to differential agglutination by peanut lectin (PNA). In the fractions obtained, the distribution and quantitative expression of CD antigens was determined by two-color fluorescence flow cytometry. A major population of CD4+8+ and minor populations of CD4+8- and CD4 8+ cells existed in unfractionated thymocytes. Agglutination of human thymocytes by PNA led to a highly effective enrichment of the CD4+8+ phenotype in the PNA+ thymocytes, and left three distinct phenotypes, CD4+8+, CD4+8-, and CD4-8+, in the PNA-thymocytes. After treatment of the CD4+8+ cells in both PNA+ and PNA- fractions for 20 hr with phorbol 12-myristate 13-acetate (PMA), 60%-90% of the cells expressed the CD4-8+ phenotype, whereas the CD4+8+ phenotype was decreased to 1%-3% of the population. In addition, pretreatment of both PNA+ and PNA- thymocytes with PMA induced suppressor activity in these cells, as shown by inhibition of immunoglobulin secretion by pokeweed mitogen (PWM)-stimulated peripheral blood lymphocytes (PBL). PMID- 2968004 TI - Plasma levels of atrial natriuretic peptide in patients with borderline and essential hypertension. AB - Plasma levels of atrial natriuretic peptide (ANP) were measured in outpatients with borderline hypertension (n = 15) and essential hypertension (n = 13) and in normotensive subject (n = 11). There were no significant differences among the three groups in age, serum protein, albumin, or electrolyte levels, plasma renin activity (PRA), or plasma concentrations of aldosterone and cortisol. The plasma ANP levels in the normotensive, borderline hypertensive, and essential hypertensive subjects were 36 +/- 6 pg/ml (mean +/- S.E.), 64 +/- 11 pg/ml, and 82 +/- 14 pg/ml, respectively. The levels in the essential hypertensive subjects were significantly (p less than 0.05) higher than those in the normotensives. In both borderline and essential hypertensives (n = 28), the plasma ANP levels were significantly correlated positively with systolic blood pressure (r = +0.385, p less than 0.05), and negatively with PRA (r = -0.484, p less than 0.05) and serum total calcium (r = -0.516, p less than 0.01). These results suggest that the elevation of circulating ANP in hypertensives is involved in the pathogenesis of hypertension. PMID- 2968005 TI - Intestinal bacterial flora and host defense mechanisms. AB - The protective mechanisms of intestinal bacterial flora against exogenous infections will be discussed in this paper. Experimental data on protective function of Escherichia coli, Enterococcus faecalis and Bacteroides distasonis comprising intestinal flora against oral infection of Shigella flexneri which causes localized infection are presented. Furthermore, the present investigation deals with non-specific defense mechanism which indicate that protective activity of intestinal flora against parenteral infection of Salmonella typhimurium mainly depend upon the larger number of functional kupffer cells in conventional mice than in that of their germfree counterparts. PMID- 2968006 TI - [Status and developmental prospects of hospital oral surgical care for children in Perm and Perm Province]. PMID- 2968007 TI - [Oral hygiene under hardest conditions]. PMID- 2968008 TI - Rapid conjugation in an extremely rapid hydroxylator of debrisoquine: a case report supporting a coregulation of certain phase I and II metabolic reactions. PMID- 2968009 TI - Differential recovery of phenotypically and functionally distinct circulating antigen-presenting cells after allogeneic marrow transplantation. AB - Low-density cells (LDC) prepared from peripheral blood by fractionation over hypertonic metrizamide contain 95% of cells with veiled morphology, almost all of which are HLA-DR-positive and have characteristics of antigen-presenting cells. In normal individuals the monoclonal antibodies RFD1 and RFD2 divide these cells into three phenotypically distinct populations, D1+D2-, D1-D2+ and D1-D2-. The RFD1-positive population is nonphagocytic. We have investigated the recovery of LDC in peripheral blood after (T cell-depleted) marrow transplantation, to assess whether defects in antigen-presenting cell (APC) subpopulations could contribute to the prolonged immune-paresis of marrow graft recipients. We find that APC of donor origin and with apparently normal morphology, phenotype, and function appear within 6 weeks of BMT. By three months the donor-derived nonphagocytic RFD1-positive subset has disappeared, although phagocytic RFD2-positive cells remain. The disappearance of the RFD1-positive subset is associated with a loss of antigen presentation by patients' LDC of the soluble protein antigen tetanus toxoid, though the capacity to present alloantigen and stimulate in a mixed lymphocyte reaction is retained. Donor-derived RFD1-positive cells and soluble antigen-presenting capacity do not reappear for one year or more. This biphasic recovery of RFD1-positive cells contrasted with the continued production of RFD2 positive APC, implies that the phenotypic and functional distinction between APC subpopulations in peripheral blood also reflects a separate ontogeny. Since these marrow graft recipients retain the phagocytic (RFD2-positive) APC but lose the nonphagocytic (RFD1-positive) APC subset, there is now an opportunity to explore the role of each subset in antigen processing and presentation. PMID- 2968010 TI - Evidence that specific cellular immunity cannot account for death of Mus caroli embryos transferred to Mus musculus with severe combined immune deficiency disease. AB - Mus caroli blastocysts transferred to pseudopregnant Mus musculus implant but die about midgestation. Death of the M caroli embryos is associated with infiltration of the fetoplacental units by M musculus lymphocytes, including functional cytotoxic T lymphocytes (CTL). To determine if specific maternal antifetal immune responses are responsible for the initiation of M caroli failure, survival of M caroli embryos transferred to M musculus with severe combined immune deficiency disease (SCID) was studied. C.B.-17 scid mice, homozygous for the scid gene, are reported to be unresponsive to T cell or B cell mitogens and to alloantigens. We have evaluated responses of C.B.-17 scid mice to M caroli xenoantigens in vivo and in vitro. C.B.-17 scid fail to reject M caroli skin grafts, and their spleen cells do not proliferate when mixed with M caroli splenocytes in a 1-way mixed leukocyte culture. Further, neither primary nor secondary challenge of C.B.-17 scid footpads with M caroli splenocytes elicits a delayed-type hypersensitivity (DTH) response. These results suggest that C.B.-17 scid mice are unable to recognize or respond specifically to M caroli xenoantigens. M caroli embryos transferred to C.B.-17 scid mice implant but fail to survive while cotransferred M musculus embryos are normal. Histologic examination of resorbing embryos revealed granulocytes, macrophages, large granular lymphocytes, but very few small lymphocytes at the implantation sites. Thus, it appears that specific maternal antifetal immune responses are not responsible for initiation of M caroli embryo failure in the M musculus uterus. These studies also indicate that efficient resorption of embryos does not require the function of T cells or B cells. PMID- 2968011 TI - An A-chain ricin immunotoxin targeted against rat class II major histocompatibility complex molecules. In vitro and in vivo effects. AB - Studies reported here have demonstrated that an antirat class II major histocompatibility complex molecule A-chain ricin immunotoxin could specifically remove, in a dose-dependent manner, the cells capable of stimulating the rat mixed lymphocyte reaction, an in vitro model of transplant rejection. It was further demonstrated that administration of a monoclonal antibody against class II major histocompatibility complex molecules to isolated rat pancreas grafts by hypothermic perfusion resulted in the targeting of cells bearing class II major histocompatibility complex molecules. But administration of the immunotoxin to isolated pancreases in a similar manner did not prolong their survival when allografted across a major histocompatibility barrier. PMID- 2968012 TI - Antigen-specific suppressor cells induced by cyclosporine. PMID- 2968013 TI - [Metronidazole cream for the local treatment of rosacea]. PMID- 2968014 TI - [Splenic rupture after laparoscopy in a patient with acute pancreatitis]. PMID- 2968015 TI - [Characteristics of free transplantation of the skin to the face]. PMID- 2968016 TI - [Organization of medical services for participants in the revolution in Petrograd in October, 1917]. PMID- 2968017 TI - [Prevention of ligature fistulas after appendectomy]. PMID- 2968018 TI - Comparison by experimental infections in cattle of a Dictyocaulus species occurring naturally in red deer and a dictyocaulus of bovine origin. AB - Dictyocaulus species larvae were obtained from young red deer which had become infected on pastures considered to be carrying the Dictyocaulus species indigenous to the red deer of Scotland. These larvae were cultured to third stage and transmitted to five bovine calves. Five other bovine calves were infected with third stage Dictyocaulus viviparus larvae of bovine origin. Microscopic appearances of both groups of larvae were indistinguishable and their lengths were similar. Results indicated that the Dictyocaulus species derived from deer induced milder though similar clinical and pathological responses in cattle than did the D viviparus derived from cattle. It was concluded that there are strains of different pathogenicity within the species D viviparus, that the deer derived Dictyocaulus species was a strain of D viviparus, and that the hazards to animal health associated with infection by D viviparus in farming systems where red deer and cattle may graze alternately are likely to be acceptable. PMID- 2968019 TI - [Organization of republican endoscopic and cytologic centers for active screening of pretumor and tumor diseases of the cervical canal and corpus uteri]. AB - An experience with screening for precancerous lesions of the cervical canal and corpus uteri is discussed. Groups at risk and high risk for cancer were formed on the basis of the results of outpatient cytologic examination of aspirates obtained from the uterine cavity through a plastic catheter. Complex endoscopic, cytologic and histologic examination of females at high risk for cancer was performed at an endoscopic center. Precancerous lesions of the endometrium and cervical canal were detected in 20% whereas cancer--in 5.2% of screened persons. PMID- 2968020 TI - [Latent viral infection and atherogenesis]. PMID- 2968021 TI - [Diagnostic possibilities of infrared thermography in viral hepatitis]. PMID- 2968022 TI - A case of occult splenic abscess following percutaneous transluminal coronary angioplasty (PTCA): an unrecognized complication of PTCA. PMID- 2968023 TI - [Global and regional left ventricular function following acute coronary artery occlusion and emergency aortocoronary venous bypass operation]. AB - Complications following percutaneous transluminal coronary angioplasty (PTCA), such as coronary artery occlusion, spasm, or dissection, frequently require urgent surgical intervention. Out of 26 patients from our hospital, 12 who underwent emergency coronary bypass grafting because of balloon catheter complications with a total ischemic time of 100-255 min, were restudied in the late postoperative period by clinical examination and by coronary and left ventricular angiography. Left ventricular angiograms were evaluated quantitatively for global and regional function using the AVD system (Siemens Elema, Erlangen). Electronmicroscopic results from myocardial biopsy samples, taken intraoperatively from the presumed center of the ischemic zone, were compared with the clinical results. An improvement of left ventricular function parameters could be seen in patients with reperfusion intervals within 130 min. In the case of intermittent perfusion of the occluded vessel (catheter perfusion with arterial blood, intermittent spasm) operative revascularization proved to salvage jeopardized myocardium. There was a close correlation between the degree of left ventricular dysfunction and histological results, coronary morphology and clinical staging. In conclusion, the efficacy of myocardial protection by emergency bypass surgery depends mainly on coronary morphology and above all on reperfusion intervals. PMID- 2968024 TI - Age-related changes in the skeletal muscle cells. AB - Skeletal muscle cells of young and old humans and CBA/Ca mice were examined electronmicroscopically. The mitochondria in old subjects and mice showed age related changes. The cristae of mitochondria became irregularly spaced, disrupted and replaced by lamellar, myelin-like structures. Giant mitochondria were often visible. They contain lipofuscin in the myofibrils, too, which was often in close relationship with the damaged mitochondria. Comparing these mitochondrial changes with those demonstrated in the lymphocytes in authors' earlier reports, it can be stated that mitochondrial changes occur more frequently in the skeletal muscle cells than in the lymphocytes. PMID- 2968026 TI - [Prevention of occupational skin diseases in workers engaged in the wet spinning of flax]. PMID- 2968025 TI - [Perthes disease--choice of therapy with reference to psychosocial aspects]. AB - Within the scape of a comparative long-term study between conservative and operative therapy of Perthes'-disease the effort was made to estimate the dimension of the psychic and social detraction in addiction to the method of treatment by a detailed inquiry of 116 patients as well as of their accompanying parents. This happened because of the opinion, that not only the formal result should decide on the choice of therapy, but also in a justifiable dimension the way of treatment should be orientated on patients. The analysis of the anamnestic particulars showed, that also under psycho-social aspects the immediate surgical procedure has the advantage over conservative methods of treatment at Perthes' disease. PMID- 2968027 TI - [Biochemical mechanism of lipidoses]. PMID- 2968028 TI - [Mental hygiene of family and marital relationships]. AB - On the basis of clinical, specialized, and sociological examination of 1771 individuals (627, 430, and 714 respectively) the authors have attempted to determine factors and tendencies which adversely affect the durability of marriage, familial and marital harmony, and mental development of children. It is concluded that from the mental hygiene standpoint and with regard to primary psychoprophylaxis of borderline diseases it is necessary to improve moral and sexual education of children and adolescents, to further develop the general education of young people about to be married, to perfect the organization of psychologic and hygienic aid to young families, to initiate scientific and methodological development of the system of lectures aimed at propagation of mental hygiene knowledge among the population. PMID- 2968029 TI - [The EEG characteristics of patients with Huntington's chorea and their clinically healthy relatives]. AB - Clinical and electroencephalographic examination has involved 170 persons (93 patients with Huntington's chorea and their 77 clinically normal relatives). The overwhelming majority of patients and 37% of their clinically healthy relatives have varying degrees of alpha-activity diminution (in 51% of the patients alpha activity was totally absent). The degree of alpha-activity significantly correlated with severity of the disease. The above EEG changes, in combination with the findings of other methods of examination, may be also employed in the diagnosis of initial and subclinical manifestations of the disease. PMID- 2968030 TI - [Signs of developmental dysplasia and function of the genetic apparatus of children and adolescents]. AB - Using scanning cytospectrophotometry and radiometry of beta-particles of 3H thymidine incorporated into DNA the authors studied chromatin in interphase nuclei of lymphocytes in children and adolescents with Down's syndrome, childhood cerebral paralysis, and undifferentiated oligophrenia, as well as in healthy subjects. The findings obtained are compared with the results of cytogenetic examination of metaphase chromosomes and the number of stigmas of dysembryogenesis. Aberrations of metaphase chromosomes were found in 4.4% of patients with childhood cerebral paralysis and oligophrenia while the pattern of changes in interphase chromatin corresponding to the chromosomal imbalance was detected in 36% of these individuals. In all cases they had from 15 to 30 stigmas of dysembryogenesis. PMID- 2968031 TI - [Characteristics of a psychopathy-like syndrome in children with oligophrenia (epidemiological research)]. AB - In an epidemiological survey of mentally retarded children of the school age the psychopathy-like syndrome was found in 16% of cases which accounted for one-third of all complicated forms of oligophrenia. Elevated affective excitability was the most common of all psychopathy-like disorders, followed by the syndrome of home leaving and tramping, the aggressive-sadistic syndrome, and mental instability. Other variants were observed far less frequently. Thirty per cent of the children presented a combination of various manifestations of the studied syndrome. Specific features of the syndromal structure were found to be correlated with the sex and age of children and degree of mental insufficiency. The correlation with antisocial behaviour was also determined. Although the diagnosis of mental retardation and transfer of such children to specialized schools were made earlier than in other groups of oligophrenic children, 4.9% of them continued their education in ordinary schools (i.e. in conditions inadequate for such children) which contributed to aggravation of their social disadaptation. PMID- 2968032 TI - [Problem of the etiology of undifferentiated oligophrenia]. AB - On the basis of examination of 150 families with clinically and etiologically undifferentiated oligophrenias the authors have studied a possible contribution of genetic and psychosocial factors to the formation of mental retardation. The examined families present a high and approximately equal frequency of mild forms of oligophrenia among parents and siblings which confirms a polygenic model of heredity. Familial and psychological factors appear to be involved in the formation of mental retardation. The data obtained may be used for predicting the status of the progeny and for developing therapeutic and corrective measures in cases of oligophrenia. PMID- 2968033 TI - [Organization of the medico-social rehabilitation of mentally retarded adolescents in specialized pediatric boarding homes]. AB - The authors present socio-hygienic characteristics of 1137 adolescents from 68 boarding homes for mentally-retarded children and clinical characteristics of 102 of these adolescents. The latter type of characterization includes the description of the degree of imbecility and attendant psychopathological disturbances. On the basis of the developed model of rehabilitation (organization of differential rehabilitation regimens, individual programme of communal and occupational adaptation of mentally retarded children, the organization of their training to acquire professional skills in specially created conditions of an industrial enterprise) the authors show a positive dynamics of attendant psychopathological disorders. It has been established that a certain proportion of adolescents with marked mental retardation may acquire professional skills and actively participate in socially useful production. PMID- 2968034 TI - Comparison of the facilitative roles of insulin and insulin-like growth factor I in the functional differentiation of granulosa cells: in vitro studies with the porcine model. AB - The facilitative effects of insulin and IGF-I were compared in vitro with regard to induction of differentiated functions of porcine granulosa cells. The monolayers were maintained under serum-free conditions in the absence or presence of porcine FSH (20 micrograms/l), with or without graded doses of insulin or IGF I. Concurrent treatment with IGF-I and FSH produced morphological differentiation and augmented LH/hCG receptor binding together with an enhancement in progesterone and estradiol secretion relative to treatment with FSH alone. IGF-I alone was incapable of exhibiting these effects. Insulin synergized with FSH to facilitate the granulosa cell functions except estradiol secretion. Maximal effective dose of IGF-I was 100 micrograms/l which is within the physiological concentration in vivo, whereas that of insulin was 1.0 mg/l, which is 1000-fold higher than the physiological level. Although the maximal effective doses of IGF I and insulin produced a comparable increment in progesterone secretion and LH/hCG receptor induction, combined treatment with IGF-I and insulin did not prove additive. [125I]IGF-I binding revealed that specific IGF-I receptors with two classes of binding sites are present on porcine granulosa cells. No distinct differences were detected between IGF-I receptors of granulosa cells from small, medium and large follicles. Insulin was approximately 100-fold less active than IGF-I in competing for [125I]IGF-I binding. These findings suggest that porcine granulosa cells possess specific IGF-I binding sites which may mediate the cytodifferentiative actions of insulin-like peptides. Since IGF-I is more potent than insulin in amplifying the actions of FSH and maximally exerts the cytodifferentiative effects at the physiological concentration, it is likely that IGF-I plays the more important role in granulosa cell differentiation in synergy with FSH. PMID- 2968036 TI - Vertebral percutaneous transluminal angioplasty. PMID- 2968035 TI - Conventional dose intravenous pulsatile GnRH therapy does not induce ovulation in polycystic ovarian disease. AB - The value of pulsatile GnRH therapy for induction of ovulation in patients with polycystic ovarian disease remains unclear. Intravenous pulsatile GnRH therapy was administered to a defined group of 5 patients with polycystic ovarian disease; all were infertile, had an LH:FSH ratio of greater than 2:1 on two or more occasions, and had multiple cysts on ovarian ultrasonography. All had failed to respond to clomiphene citrate. The 5 patients received increasing doses of GnRH (5-40 micrograms/pulse) continuously for up to 6 weeks. The response was evaluated by serial hormone levels and ovarian ultrasonography. During nine treatment periods no patient ovulated, and in only one did the LH:FSH ratio revert to normal. Four patients have subsequently had wedge resection of the ovaries and in each case the diagnosis of polycystic ovarian disease was confirmed. Pulsatile GnRH therapy was of no value in the management of this group of infertile patients with strictly defined polycystic ovarian disease. PMID- 2968037 TI - Regulator soluble factors of human IgE synthesis. I. Production and isolation. AB - The system of IgE response undoubtedly has a special interest, given the role of this immunoglobulin on the pathogenesis of allergic diseases. In the last few years the existence of a regulator mechanism that maintains the synthesis of IgE in low but effective levels has been confirmed. This mechanism consists in the equilibrium between the actions of suppressors and enhancers. These two soluble factors present in the serum and capable of suppressing and enhancing IgE response in control rats have been identified; both have a highly selective activity on IgE, exerting their effect in an antigen-independent form and require specie-specificity. Motivated by these findings, we have applied this experimental system used in rats to humans. Healthy controls were selected as low responders of IgE, atopics as high responders, and asthmatic pollinosis considered as the prototype of allergic disease mediated by IgE. After obtaining the factors mentioned by means of a bi-directional mixed culture in a pool of lymphocytes, we proceeded to their isolation by affinity chromatography in Con-A Sepharose columns. The glycoproteins obtained were afterwards submitted to different techniques of characterization: immunoelectrophoresis in polycrilamide gel, chromatography technique, etc. PMID- 2968038 TI - Cardiac involvement in hypertension. AB - Despite improved patient detection and pharmacologic therapy, the effect of treatment of hypertension on mortality from coronary artery-related events remains unresolved. Left ventricular (LV) hypertrophy, a known consequence of hypertension, is associated with an excess mortality independent of other known cardiovascular risk factors. Recently, LV hypertrophy accompanying hypertension has been associated with ominous ventricular arrhythmias. However, it does not necessarily follow that regression of LV hypertrophy will reduce this increased mortality. Diastolic dysfunction, manifested by reduced ventricular distensibility of the hypertrophying left ventricle, appears to be an early characteristic of the hypertensive heart since echocardiographic techniques have demonstrated diastolic filling abnormalities in untreated essential hypertensives even before significant LV hypertrophy appears. Not all antihypertensive agents diminish LV mass and improve diastolic dysfunction. Certain sympatholytic agents, calcium antagonists, beta-adrenergic blockers, and the angiotensin-converting enzyme inhibitors appear to diminish LV hypertrophy. However, future studies are needed to determine if these agents that appear to reverse findings of LV hypertrophy and improve diastolic dysfunction will also reduce risk of coronary artery disease and related events. PMID- 2968040 TI - Survival to age 65 years with tricuspid and pulmonic valve atresia. PMID- 2968039 TI - Sensitivity and specificity of electrocardiographic criteria for left and right ventricular hypertrophy in morbid obesity. AB - To determine the sensitivity and specificity of standard electrocardiographic criteria for left ventricular (LV) and right ventricular (RV) hypertrophy in morbid obesity, resting electrocardiograms and M-mode echocardiograms were obtained in 65 patients whose actual body weight was more than twice their ideal body weight and who were free from hypertension and organic heart disease not directly attributable to obesity. Electrocardiographic criteria for LV hypertrophy were tested using increased LV wall thickness, LV enlargement and increased LV mass (all determined echocardiographically) as diagnostic standards. Electrocardiographic criteria for RV hypertrophy were tested using echocardiographic RV enlargement or RV hypertrophy as a diagnostic standard. Sensitivity values for the electrocardiographic criteria for LV hypertrophy ranged from 0 to 13%, 0 to 20% and 0 to 12% using echocardiographic increased LV wall thickness, LV enlargement and increased LV mass, respectively, as diagnostic standards. Specificity values ranged from 73 to 100%, 87 to 100% and 83 to 100%, respectively, using these diagnostic standards. Sensitivity values for the electrocardiographic criteria for RV hypertrophy ranged from 0 to 16% and specificity values ranged from 95 to 100%. Combining electrocardiographic criteria within groups did not appreciably increase sensitivity and often decreased specificity to unacceptably low levels. The electrocardiogram is very limited in its ability to detect ventricular hypertrophy and chamber enlargement in morbidly obese patients. PMID- 2968041 TI - Effects of percutaneous transluminal coronary angioplasty on atherosclerotic plaques and relation of plaque composition and arterial size to outcome. AB - To delineate their relation to outcome of percutaneous transluminal coronary angioplasty (PTCA), the atherosclerotic plaque composition and coronary artery size in 82 five-mm long segments at 28 PTCA sites were determined in 26 patients having PTCA. The 26 patients were subdivided into 3 groups according to the degree of angiographic patency at the end of the PTCA procedure and to the duration of survival after PTCA (less than or equal to 30 or greater than 30 days): early success (13 patients, 16 PTCA sites and 49 five-mm segments); early failure (4 patients, 4 PTCA sites and 16 five-mm segments) and late success (9 patients, 8 PTCA sites and 17 five-mm segments). The mean percent of plaque comprised of fibrous tissue among the 3 groups was 80 +/- 18%, 71 +/- 23% and 82 +/- 16% (difference not significant); the mean percent of plaque comprised of lipid was 17 +/- 16%, 21 +/- 24% and 16 +/- 15% (difference not significant); and of calcium it was 3 +/- 4%, 8 +/- 10% and 2 +/- 3% (p = 0.01). The mean coronary arterial internal diameter was 3.3 +/- 0.6, 3.9 +/- 1.2 and 3.2 +/- 0.7 mm (p less than 0.02). Plaque tear was present in 1 or more histologic sections in 25 of the 26 patients and the 1 patient without it had the longest interval (nearly 3 years) between PTCA and death. Plaque tear extending from intima into media with dissection was observed only in the early and late success groups (p = 0.03). Hemorrhage into plaque was present in 16 (80%) of 20 PTCA sites in the 2 early groups and in 3 (37%) of 8 sites in the late group (p less than 0.03). Occlusive thrombus (5 of 16, 1 of 4 and 1 of 8) and plaque debris (7 of 16, 1 of 4 and 2 of 8) in residual lumens were insignificantly different among the 3 groups and their 82 five-mm segments. Plaques that had greater than 25% lipid content, however, had an increased frequency of hemorrhage into plaque (p less than 0.004), occlusive thrombus (p = 0.0001) and plaque debris in residual lumens (p less than 0.05). These findings suggest that coronary arterial size and plaque composition are strong determinants of PTCA outcome. The ideal coronary arterial atherosclerotic narrowing for both technically and clinically successful PTCA appears to be a small (less than 3.3 mm in internal diameter) artery in which the plaque contains relatively little calcium and lipid. PMID- 2968042 TI - Estimation of the secretion rate of atrial natriuretic peptide from the coronary sinus in coronary artery disease. AB - Although atrial natriuretic peptide (ANP) is known to be secreted through the coronary sinus into the systemic circulation, its actual secretion rate has not been thoroughly investigated. The immunoreactive ANP concentrations in plasma samples from the ascending aorta and coronary sinus in 11 patients with the coronary artery disease were measured and the coronary sinus flow rate using the continuous thermodilution method was simultaneously determined at the time of sampling. These variables were also determined during the intravenous infusion of synthetic alpha-human ANP at 0.025 microgram/kg.min in 7 of the 11 patients. In the basal state, the plasma concentration of ANP was 61 +/- 6 (standard error) pg/ml in the aorta and 541 +/- 40 pg/ml in the coronary sinus, and the coronary sinus flow index was 57.3 +/- 12.3 ml/min.m2. Thus, the secretion rate of ANP was determined to be 14.4 +/- 2.8 ng/min.m2. The secretion rate of ANP correlated significantly with the plasma concentration of ANP in the aorta (r = 0.65, p less than 0.05). The ANP infusion, which decreased pulmonary artery wedge pressure from 8.0 +/- 0.6 to 6.3 +/- 0.4 mm Hg (p less than 0.01), elevated the plasma concentrations of ANP in the aorta and coronary sinus by 701% (p less than 0.001) and 33% (p less than 0.05), respectively, and decreased the secretion rate of ANP by 40% (p less than 0.05). These results suggest that the circulating plasma concentration of ANP may reflect the secretion rate of ANP and that an increase in circulating ANP directly or indirectly reduces ANP secretion. PMID- 2968043 TI - Electrocardiographic estimate of left ventricular mass versus radiographic cardiac size and the risk of cardiovascular disease mortality in the epidemiologic follow-up study of the First National Health and Nutrition Examination Survey. AB - The prognostic value of a left ventricular (LV) mass index (g/m2) estimated from an electrocardiographic model and radiographic estimates of the relative heart volume (ml/m2) and cardiothoracic ratio for predicting the risk of cardiovascular disease mortality were investigated using Cox regression analysis to adjust for age, systolic blood pressure and history of heart attack in 1,807 men (1,609 white, 198 black) and 2,143 women (1,884 white, 259 black). The study population (ages 35 to 74 years at baseline) was followed from 5 to 12 years (average 9.5 years) for cardiovascular disease mortality. LV mass index and relative heart volume were independent predictors of cardiovascular disease mortality among white men. All 3 cardiac size estimates were independent predictors for cardiovascular disease mortality among white and black women. When LV mass index was used as a dichotomized variable to indicate the presence or absence of LV hypertrophy, the age-adjusted relative risk of cardiovascular disease mortality was 2.48 (95% confidence interval 1.77 to 3.46) for white men, 3.03 (1.49 to 6.16) for black men, 1.86 (1.21 to 2.87) for white women and 2.05 (0.83 to 5.05) for black women. The corresponding prevalence of LV hypertrophy was 15.4% for white men, 36.6% for black men, 20.1% for white women and 17.4% for black women. It is concluded that the electrocardiographic estimate of LV mass index can identify a substantially larger fraction of persons at increased risk for cardiovascular mortality than conventional electrocardiographic criteria for LV hypertrophy and that LV mass index estimated by electrocardiogram is a valuable supplement to radiographic cardiac size estimates in epidemiologic applications. PMID- 2968044 TI - Hepatitis B virus infection in Danish dentists. A case-control and follow-up study. AB - The authors determined the seroprevalence of hepatitis B virus markers among Danish dentists in 1985 in an effort to assess occupational risk factors. A case control study approach was applied using pedodontists as zero-exposure controls. The study further allowed serologic and epidemiologic follow-up of dentists who participated in a similar study in 1976. The authors sought to determine whether exposure to hepatitis B virus in this population sometimes leads to immunization without infection (antibody to hepatitis B surface antigen (anti-HBs) positivity only). A total of 922 dentists (22% of all Danish dentists) gave blood; of these, 254 (28%) participated in the 1976 study. Serum samples were tested using commercial solid phase radioimmunoassays. The prevalences of anti-HBs and antibody to hepatitis B core antigen (anti-HBc) were 7.2% (95% confidence interval (CI) = 5.7-9.0) and 5.2% (95% CI = 3.8-6.6), respectively. Twenty-one dentists (31%) only had anti-HBs (without prior vaccination). Age-standardization of the two data sets (1976 and 1985) indicated no substantial change in seroprevalence. Private practitioners, especially those working in Copenhagen, had an increased odds ratio (OR) of being seropositive compared with that of pedodontists (OR = 3.0 (95% CI = 1.3-6.8) adjusted for age and sex and OR = 2.4 (95% CI = 1.1-5.2) adjusted for age and site of practice). The proportion attributable to occupational exposure was approximately 50%. In a logistic regression model, site of practice and type of specialty were associated with hepatitis B virus seropositivity (p = 0.01 and 0.05, respectively), whereas age, sex, hours of oral surgery per week, and use of gloves were not. Four persons had become seropositive in the follow-up study, for a seroconversion rate of 0.2% per year. Seven out of 18 dentists who were anti-HBs positive in 1976 were found to be anti-HBs negative in 1985. Most of these dentists had values below 10.0 units by S/N ratio (ratio of counts per minute of sample compared with the mean number of counts per minute of negative controls). The findings do not justify institution of a large-scale vaccination program for dentists, but there are certain subgroups of dentists which constitute candidates for vaccination. PMID- 2968045 TI - Fetal cardiac Doppler flow studies in prenatal diagnosis of heart disease. AB - The prenatal diagnosis of fetal cardiac disease has become increasingly accurate as the technology of ultrasound has improved. Although two-dimensional real-time ultrasound remains the primary method of diagnosis, Doppler blood flow velocity estimates can provide valuable pathophysiologic information to support the anatomic diagnosis. We present six cases in which Doppler studies contributed to the accuracy of the diagnosis of fetal heart disease, including tetralogy of Fallot, right and left ventricular hypoplasia, atrioventricular canal defect, double-outlet right ventricle, and pulmonic stenosis. Velocities in these cases are compared with those in normal fetuses. If Doppler flow velocities are not consistent with the observed morphologic changes, further observations are indicated. Inasmuch as most anatomical heart lesions result in altered flow patterns, Doppler investigations of intracardiac and extracardiac flow should be a routine component of the fetal echocardiogram when structural abnormalities are found. PMID- 2968046 TI - The predictability of the small-for-gestational-age infant by real-time ultrasound-derived measurements combined with pulsed Doppler umbilical artery velocimetry. AB - During a 15-month period 373 level II ultrasound examinations were performed in 256 high-risk patients. In addition, pulsed Doppler spectral recordings of blood flow in the fetal umbilical arteries were made. A systolic/diastolic ratio was then calculated for each fetus. Real-time ultrasound-derived estimated fetal weight with the use of biparietal diameter and abdominal circumference was also calculated. The estimated fetal weights were categorized by placing them in a percentile for gestational age according to published nomograms. Complete birth data and outcomes were obtained in all patients. Both the systolic/diastolic ratio and ultrasound-estimated fetal weight grouped by percentile ranking for gestational age were highly predictive (p = 0.001) of babies who were subsequently born small for gestational age. Seventy-nine percent of the infants small for gestational age had umbilical artery systolic/diastolic ratios greater than or equal to 4, whereas only 21% had normal systolic/diastolic ratios. Forty three percent of the infants who were small for gestational age had ultrasound estimated fetal weights less than or equal to 10th percentile for the gestational age at which it was measured. Umbilical artery systolic/diastolic ratios, which reflect an increase in peripheral resistance in the placental circulation, showed a highly predictive and discriminatory index for the evaluation of the fetus suspected of having growth retardation. PMID- 2968047 TI - Systemic immunosuppression induced by peritoneal photodynamic therapy. AB - Although photodynamic therapy is being used increasingly for the diagnosis and treatment of human cancer, its effect on immune responses has received little attention. This aspect was examined in a murine model. Mice given peritoneal photodynamic therapy had markedly decreased contact hypersensitivity responsiveness, not observed with cutaneous photodynamic therapy. The immunosuppression was systemic, because contact hypersensitivity was depressed at distal, unirradiated sites. Photodynamic therapy induced adoptively transferable cells that inhibited contact hypersensitivity responses in naive mice. The immunosuppression was reversible, but persisted for 3 weeks after photodynamic therapy. An acute-phase response characterized by leukocytosis and elevated serum amyloid P levels was observed in mice given photodynamic therapy but not in mice treated with either laser or dye alone. These data suggest a link between the acute-phase response and immunosuppression. Thus, although photodynamic therapy shows promise in cancer treatment, the induction of decreased systemic immunoresponsiveness is an important observation with potentially detrimental consequences. PMID- 2968048 TI - Effect of chlorpromazine on the development of experimental glomerulonephritis and Arthus reaction. AB - Chlorpromazine blocks antibody-mediated redistribution of cell surface antigens in vitro and in vivo and inhibits the development of passive Heymann glomerulonephritis, a disease characterized by in situ formation of immune complexes (Camussi et al J Immunol 1986, 136:2127-2135). The aim of this study was to establish whether chlorpromazine exerts similar effects in other rat models characterized by in situ formation of immune complexes. In glomerulonephritis induced by antibodies reactive with an exogenous antigen "planted" in glomeruli pretreatment with chlorpromazine prevented formation of "humps" and exudative and proliferative lesions. Likewise, chlorpromazine prevented passive reverse Arthus reaction in the skin. In contrast, the drug was ineffective when these lesions were already established, and also failed to inhibit the fulminant course of nephrotoxic serum glomerulonephritis with an enhanced autologous phase. It is proposed that the antiinflammatory effect of chlorpromazine is due to its ability to block the recruitment of inflammatory cells and the release of inflammatory mediators. PMID- 2968049 TI - Antigenic charge as a factor in resistance to immunosuppressive therapy. AB - This study examines the effects of methylprednisolone (MP) on the glomerular lesions induced by the injection of cationic bovine serum albumin (BSA) in rabbits. Male New Zealand white rabbits (n = 55) were treated with cationic BSA alone (n = 13), cationic BSA plus MP (n = 26), native BSA alone (n = 8), or native BSA plus MP (n = 8). Animals receiving BSA alone developed subepithelial immune complex deposits after injections of cationic BSA or mesangial immune complex deposits after injections of native BSA. Subepithelial deposits were inhibited in only 10 of 26 rabbits receiving MP and cationic BSA. Glomerular lesions occurred rather predictably in this group unless complete ablation of antibody production was achieved. Even minimal amounts of detectable antibody were sufficient to support the development of the full-blown renal lesion. Proteinuria was diminished in all animals receiving cationic BSA and MP in whom a beneficial histologic effect was observed. MP inhibited the development of mesangial deposits in all of the animals given native BSA. This inhibition was associated with almost complete ablation of anti-BSA antibody production in this group. Renal perfusion studies (n = 8) demonstrated that the beneficial effects of MP on the development of immune deposits were related to systemic immunosuppression and not to local structural alterations in the glomerulus. Our findings confirm that the pattern of immune response to native and cationic antigens differ. In addition, they demonstrate that cationic antigens appear to require only small amounts of free circulating antibody in order to produce glomerular immune complexes and cause proteinuria. PMID- 2968050 TI - The immunopathology of acute experimental allergic encephalomyelitis. V. A light microscopic and ultrastructural immunohistochemical analysis of fibronectin and fibrinogen. AB - Cellular expression and extracellular deposition of fibronectin and fibrin/fibrinogen in experimental allergic encephalomyelitis (EAE) in guinea pigs (GP) were studied by light and electron microscopic immunohistochemistry of the central nervous system (CNS). Normal GPs had few fibronectin+ and fibrin/fibrinogen+ CNS vessels. Positively stained vessels were more numberous in sensitized GPs prior to and after onset of EAE, and there were extracellular deposits of these molecules in inflammatory foci. Staining was greater in EAE susceptible strain 13 GPs than in sensitized EAE-resistant strain 2 GP. On ultrastructural analysis both molecules were localized on endothelial cell luminal surfaces and on intravascular mononuclear cells. Fibronectin and the macrophage fibronectin receptor (A6F10) were found on mononuclear cells in parenchyma. Endothelial cell luminal surface fibronectin and fibrin/fibrinogen may mark and/or contribute to early immune events in EAE, including mononuclear cell margination and emigration in inflammatory sites. Genetic factors, cytokines, and components of coagulation may regulate these interactions in vivo. PMID- 2968051 TI - Hemodynamic and renal responses to volume expansion in dogs with cardiac denervation. AB - Hemodynamic responses, renal function, and plasma levels of immunoreactive atrial natriuretic factor (irANF) were examined following volume expansion (VE) in normal (N) conscious dogs and in conscious dogs with cardiac denervation (CD). Base-line urine flow was consistently greater (P less than 0.05) in dogs with CD (0.54 +/- 0.06 ml/min) than in N (0.29 +/- 0.03 ml/min) dogs but sodium excretion did not differ between N (2.80 +/- 0.58 mu eq.min-1.kg body wt-1) and CD (3.53 +/ 0.75 mu eq.min-1.kg-1) groups. With VE (18 ml/kg of 3% dextran in saline), mean arterial pressure (MAP) increased (P less than 0.01) by 16 +/- 3 from 103 +/- 4 mmHg in N dogs but did not change from pre-VE base line (103 +/- 2 mmHg) in dogs with CD. At 10 min after VE, urine flow increased more (P less than 0.01) in N dogs (1.39 +/- 0.24 ml/min) than in dogs with CD (0.26 +/- 0.09 ml/min). At that time, increases in sodium excretion were also greater (P less than 0.01) in N (9.13 +/- 1.96 mu eq.min-1.kg-1) dogs than in dogs with CD (1.06 +/- 0.68 mu eq.min-1.kg-1). With VE, increases in irANF plasma levels were not different in N dogs (40 +/- 12 from 34 +/- 5 pg/ml) and in dogs with CD (27 +/- 3 from 45 +/- 7 pg/ml). In dogs with CD, when MAP was increased by aortic constriction to mimic responses observed in N dogs, renal responses were similar to those of N dogs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968053 TI - Right ventricular hypertrophy long after reversal of severe pressure overload in cats. AB - To test the hypothesis that alterations in ventricular size and function may persist long after reversal of a severe pressure overload, right ventricular (RV) hypertrophy was produced in cats by abrupt, severe constriction of the pulmonary artery (PA) for 8 wk, after which pressure overload was completely reversed by surgical removal of the PA band (group III). Despite debanding and recovery from pressure overload (RV peak systolic pressure = 22 +/- 4 vs. 66 +/- 20 mmHg before debanding and 25 +/- 4 mmHg in controls), parameters of hypertrophy 8-10 mo later remained substantially increased: e.g., RV/LV (left ventricular) mass ratio was 0.36 +/- 0.06 in group III vs. 0.24 +/- 0.03 in controls (group I) and 0.43 +/- 0.07 in a separate cohort (group II) studied after 8 wk of banding. Maximum RV stroke work during volume loading was computed from RV micromanometry and PA electromagnetic flow: 20 +/- 8 g-cm/g for group III vs. 51 +/- 14 for group I, P less than 0.05, and 21 +/- 7 for group II. Thus severe pressure overload of the RV may produce changes in size and function that are not fully reversed by complete removal of the overload. PMID- 2968052 TI - Plasma clearance and effects of alpha-hANP infused in patients with end-stage renal failure. AB - The effects and clearance of synthetic atrial natriuretic peptide (alpha-hANP) were investigated in eight patients with end-stage renal failure and six normal volunteers. ANP or vehicle was infused for 1 h at 10 pmol.kg-1.min-1 in random order on two separate occasions. During ANP infusions in end-stage renal patients, microhematocrit rose by 9.8 +/- 2% (P less than 0.005, n = 8), from base-line values of 0.24 +/- 0.02. Serum protein and albumin rose consistently. In contrast, during placebo infusions, no significant changes were seen. Blood pressure, heart rate, plasma renin concentration, serum creatinine, and electrolytes did not change significantly during either study phase. In end-stage renal failure patients, metabolic clearance rate of infused ANP was 1.04 +/- 0.095 l/min and its plasma half-life was 4 min 34 s. In normal volunteers, metabolic clearance rate was 2.6 l/min and its plasma half-life 3 min 30 s. The data suggest that ANP promotes contraction of plasma volume via a mechanism independent of renal function and also indicate that the kidney is not the only organ involved in the ANP metabolism. PMID- 2968054 TI - Aldosterone and renin inhibition by physiological levels of atrial natriuretic factor. AB - This study was designed to examine, in the rat, the inhibition of renin release and aldosterone secretion by physiological plasma levels of atrial natriuretic factor (ANF). Intravascular volume expansion over 30 min with donor blood equal to 3% body weight increased plasma ANF concentration from a base line of 216 +/- 28 to 1,590 +/- 240 pg/ml (P less than 0.001) in sodium-replete rats. Basal plasma ANF levels were decreased to 125 +/- 9 pg/ml in animals fed a low-sodium diet, and the infusion of synthetic ANF into separate groups of these rats at doses of 15 or 45 ng.kg-1.min-1 elevated plasma ANF to 346 +/- 38 and 720 +/- 96 pg/ml, respectively (P less than 0.001 for both values). Infusion of ANF at 15 ng.kg-1.min-1 resulted in a significantly lower plasma renin activity [30 +/- 3 vs. 69 +/- 5 ng angiotensin I (ANG I).ml-1.h-1, P less than 0.05], but there was no difference in aldosterone secretion between control and infused groups (7.00 +/- 0.49 vs. 7.29 +/- 0.95 ng/min, P greater than 0.05). However, the higher ANF dose of 45 ng.kg-1.min-1 did reduce aldosterone secretion approximately 40% to 4.18 +/- 0.36 ng/min (P less than 0.001) with no further suppression in plasma renin activity. In sodium-replete rats, infusion of ANF at 45 ng.kg-1.min-1 resulted in lower plasma renin activity compared with the control noninfused group (14 +/- 4 vs. 22 +/- 2 ng ANG I.ml-1.h-1, P less than 0.05), but aldosterone secretion was not different (P greater than 0.05) between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968055 TI - Renal and endocrine responses in the elderly during head-out water immersion. AB - After overnight food and fluid restriction, seven healthy old (62-74 yr) and eight young (21-28 yr) men were examined before, during, and after 3-h head-out immersion (HOI) in thermoneutral water (34.5 +/- 0.5 degrees C). On separate days, all subjects remained seated in air for 5 h to obtain the time control data. Although HOI induced a reversible increase in urine flow in all subjects, the response was faster and greater in magnitude in the elderly than in the young. Na excretion and osmolal clearance also followed a response pattern identical to that of urine flow; thus the HOI-induced diuresis was entirely osmotic. Endogenous creatinine clearance increased in the elderly at 2 h of HOI, suggesting an age-related modification in kidney hemodynamics. Although there was virtually the same cephalad blood shift (measured by impedance cardiography), mean arterial pressure significantly increased (P less than 0.05) during HOI in the elderly, which also indicated a different response of peripheral circulation to HOI in the elderly. Control level of plasma atrial natriuretic factor (ANF) was nearly twofold greater in the elderly compared with the young. The HOI induced a nearly fourfold increase in ANF in the elderly, whereas that for the young was threefold. Both plasma aldosterone and ADH responses to HOI were attenuated in the elderly compared with the young, which had no correlation with urine flow or Na excretion. It is concluded that the elderly release more ANF at a given cephalad volume expansion compared with the young, but the vasodilative reaction to ANF was attenuated in the elderly. PMID- 2968056 TI - An overview of Japanese occupational health. AB - This paper provides an overview of Japanese occupational health and evaluates the current situation from three perspectives. Major occupational health hazards are assessed using four sources of data, showing patterns similar to those found in other advanced industrial societies. Institutional structures for occupational health policy are then examined, illustrating strengths and weaknesses of the Japanese legal and administrative systems. Trade union activities are presented, indicating the constraints of enterprise unions, and the tendency for a greater orientation toward compensation than prevention. Significant occupational health problems persist among marginal workers in Japan, including women and various minority groups. The analysis demonstrates a record for occupational health in Japan considerably more mixed than the conventional view. PMID- 2968057 TI - A microassay for ATPase. AB - A newly developed microtechnique for quantitating activity of myosin ATPase (EC 3.6.1.32) is more sensitive and less time-consuming than existing spectrophotometric methods. Measurement of ATPase activity using the new method can be accomplished in a final volume of 0.25 ml, allowing the assay to be conducted in individual wells of 96-well microplates commonly used for the enzyme linked immunosorbent assay (ELISA). The microassay is performed by adding purified myosin to microplate wells followed by addition of ATP to initiate the enzymatic reaction. The reaction is subsequently terminated by addition of an acidic solution containing malachite green and ammonium molybdate. The level of inorganic phosphate produced by enzymatic hydrolysis of ATP is measured by scanning the microplates using a microELISA plate reader. An entire 96-well microplate can be scanned in less than 2 min, and data from the microassay can be transferred directly to a microprocessor for statistical analysis. The microassay is capable of detecting between 0.2 and 3 nmol of inorganic phosphate in a reaction volume of 50 microliter, and the ATPase activity of as little as 10 ng of rat cardiac myosin can be measured. The increased sensitivity compared with that of other spectrophotometric assays and ease of performing the microassay enable a detailed analysis of the enzymatic properties of cardiac myosin to be conducted on large numbers of small tissue specimens. Several kinetic properties of rat cardiac myosin were determined using this technique. PMID- 2968059 TI - Possible link between skin infections and HIV. PMID- 2968058 TI - Effect of hypothermic hemodilutional cardiopulmonary bypass on plasma sufentanil and catecholamine concentrations in humans. AB - The effect of hypothermic hemodilutional cardiopulmonary bypass (CPB) on plasma sufentanil and catecholamine concentrations was studied in four groups of ten patients each, receiving four different doses of sufentanil. Samples for measurement of sufentanil were obtained before CPB, at 15, 30, and 45 minutes of CPB, during rewarming, immediately after and 15, 60, and 240 minutes after CPB. In addition, in groups III and IV, which received the highest dose of sufentanil, blood samples were also obtained for measurement of plasma levels of epinephrine and norepinephrine. Sufentanil concentration decreased in all groups with the start of CPB (group I, 2.92 +/- 0.2 to 2.04 +/- 0.2; group II, 3.30 +/- 0.3 to 1.51 +/- 0.2; group III, 7.08 +/- 0.7 to 3.45 +/- 0.3; group IV, 10.33 +/- 0.5 to 4.59 +/- 0.5 ng/ml). No further decreases occurred during CPB but increases occurred with rewarming. The first measurement after CPB approached the concentration before CPB (group I, 2.82 +/- 0.3; group II, 2.56 +/- 0.5; group III, 4.42 +/- 0.4; group IV, 6.10 +/- 0.4 ng/ml). Norepinephrine concentrations demonstrated a wide variability with no significant changes. Epinephrine levels increased significantly during rewarming in both groups (group III, 141 +/- 23 to 279 +/- 79 pg/ml; P less than 0.05; group IV, 105 +/- 24 to 267 +/- 68 pg/ml, P less than 0.05). The stability of plasma sufentanil concentrations during CPB suggest that no measurable metabolism or excretion occurred. The increase with rewarming and after CPB suggest significant sequestration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968060 TI - Effects of loratadine (SCH 29851) in suppression of histamine-induced skin wheals. AB - The efficacy and safety of single oral doses (10, 20, 40, and 80 mg) of loratadine (SCH 29851) in suppressing formation of histamine-induced wheals were assessed in a crossover study in 29 healthy male subjects. One hour prior to dosing and 1, 2, 3, 4, 6, 8, 12, 16, 24, 28, 32, 36, 40, and 48 hours after dosing, histamine and saline were injected intradermally into opposite arms. Measurements of resulting wheal areas showed loratadine suppressed wheal formation significantly better than placebo; suppression was dose related. The mean suppression over 48 hours was 16% in placebo-treated subjects and 35%, 45%, 51%, and 67% in the 10, 20, 40, and 80 mg loratadine-treated subjects, respectively. The onset of action occurred within the first hour. Duration of suppression was dose related, ranging from 12 hours with the lowest dose (10 mg) to 48 hours with the higher doses (40 and 80 mg). Incidence of sedation and other side effects were comparable among all doses of loratadine and placebo. PMID- 2968062 TI - Myths and pitfalls in emergency translaryngeal ventilation: correcting misimpressions. AB - Translaryngeal jet ventilation has been proven an effective emergency airway alternative. However, confusion exists as to the proper cannulae and oxygen sources for this technique. Our study was designed to determine the delivered volumes of gas using cannulae and oxygen sources recommended in previous reports on translaryngeal jet ventilation. From this, we hope to clarify the proper technique of translaryngeal jet ventilation. Using a variety of oxygen sources and cannulae, peak flow rates were measured using a digital flowmeter. Delivered volumes of gases generated with each combination were then calculated. All of the cannulae tested (standard 16-gauge IV cannulae and larger) provided peak flow rates high enough so that predicted tidal volumes would be adequate to maintain adequate ventilation in apneic adults when a 50-psi source was used. Only a 4-mm tracheal cannula provided comparable values when a bag-valve device was used. No cannulae provided sufficient flow rates to ensure adequate ventilation in apneic adults when a demand-valve mechanism of 60 cm H2O driving pressure served as the source. Our observations were consistent with previous clinical studies and suggest that standard translaryngeal jet ventilation cannulae (12 to 16 gauge) must be connected to an oxygen source of 50 psi in apneic adults. Demand-valve devices do not provide sufficient driving pressures for these cannulae. A cannula of 4 mm ID should be placed if only a bag-valve device is available for ventilation. PMID- 2968061 TI - Cytoprotective drugs: a new perspective in the treatment of adverse reactions to foods. AB - Food allergy (FA) and food intolerance (FI) are complex syndromes caused by adverse reactions to foods. Since mucosal permeability of the digestive tract is often increased during this pathology, we evaluated the clinical efficacy of two different cytoprotective drugs in patients with urticaria-angioedema due to FA and FI. These drugs were pirenzepine, an anti-muscarinic (anti-MI) receptor antagonist, and rosaprostol, a synthetic prostaglandin. Further, the results obtained by these schedules of treatment were compared with those achieved by the previously described polyantihistaminic treatment (ie, the association of anti-H1 plus anti-H2 receptor blockers). The present investigation suggests that the cytoprotective drugs are more effective than the antisecretive ones (ie, the anti H2). Recently, anti-H2 drugs and ketotifen were shown to be in vitro inhibitors of the immune response and cromolyn was demonstrated capable of exerting an enhancing effect on T cell proliferation. In the present study we tested the effect of pirenzepine on several immunologic parameters, such as lymphocyte proliferation (through different activation pathways) and lymphokine (interleukin 2 and interferon-gamma) production. Since we found that pirenzepine does not affect the immune response and in consideration of its clinical efficacy, we consider this cytoprotective drug a valuable tool in the treatment of adverse reactions to foods. PMID- 2968063 TI - Pain and suffering. A reappraisal. PMID- 2968064 TI - Antibody to human immunodeficiency virus (HIV) and suboptimal response to hepatitis B vaccination. AB - STUDY OBJECTIVE: To analyze the relation between human immunodeficiency virus (HIV) infection and the antibody response to plasma-derived hepatitis B vaccine. DESIGN: Open-label longitudinal cohort study; blinded laboratory studies. SETTING: University-affiliated municipal hospital. PATIENTS: Homosexually active men with negative assays for hepatitis B surface antigen (HBsAg), hepatitis B core antigen, and antibody to HBsAg; recruited in a sexually transmitted disease clinic or referred from community practitioners. INTERVENTIONS: Immunization with 20 micrograms of plasma-derived hepatitis B virus vaccine intramuscularly, repeated after 1 and 6 months; standardized evaluation at entry and at 1, 2, 6, and 7 months. MEASUREMENTS AND MAIN RESULTS: Low antibody response or nonresponse to vaccination occurred in 7 of 16 HIV-seropositive patients, compared with 6 of 68 HIV-seronegative patients (P = 0.002). Median levels of antibody to HBsAg 7 months after the first vaccine dose were 205.3 sample ratio units for HIV seronegative patients and 15.5 sample ratio units for HIV-seropositive patients. By multivariate analysis, vaccine response was associated with HIV antibody status and not with cytomegalovirus infection, lymphocyte subset results, or impaired cutaneous delayed hypersensitivity. CONCLUSIONS: Infection with HIV is associated with suboptimal antibody response to plasma-derived hepatitis B virus vaccine. Determination of antibody levels after vaccination in HIV-seropositive patients may be warranted. PMID- 2968065 TI - Hepatitis B prevention and human immunodeficiency virus (HIV) infection. PMID- 2968066 TI - Molecular mechanism of regulation of phospholipid metabolism in membranes. PMID- 2968067 TI - Antenatal detection of placental steroid sulphatase deficiency by measurement of urinary 16 alpha-hydroxydehydroepiandrosterone sulphate. AB - A simple gas chromatographic technique for the measurement of 16 alpha hydroxydehydroepiandrosterone sulphate in urine from pregnant women is described. An assessment was made of the effectiveness of the measurement of this oestriol precursor for the antenatal diagnosis of placental steroid sulphatase deficiency. Twenty-two patients whose pregnancies were complicated by subnormal oestrogen excretion for gestation were studied. In nine of these, where placental steroid sulphatase activity was found subsequently in vitro to be normal, the excretion of 16 alpha-hydroxydehydroepiandrosterone sulphate was less than 27 mumol/24 h. In the remaining 13 patients, in whom postnatal in vitro assay demonstrated absence of placental steroid sulphatase activity, urinary excretion of 16 alpha hydroxydehydroepiandrosterone sulphate was 59-360 mumol/24 h. The excretion of this metabolite was below the limit of detection (20 mumol/24 h) in 30 uncomplicated pregnancies. It is concluded that urinary excretion of 16 alpha hydroxydehydroepiandrosterone sulphate greater than 50 mumol/day or a ratio of urinary 16 alpha-hydroxydehydroepiandrosterone sulphate to urinary oestrogen greater than 2.0 correctly identifies, before delivery, those pregnancies in which fetus and placenta are deficient in steroid sulphatase activity. PMID- 2968068 TI - Control of cerebral blood flow. PMID- 2968069 TI - Symptomatic atlantoaxial dislocation in Down's syndrome. PMID- 2968070 TI - [Clinical evaluation of BRMs in gastric cancer patients]. AB - The concept of BRM includes both immunotherapeutic agents and cytokines. We have tested the effects of various kinds of immunopotentiators on gastric cancer patients in terms of (1) the effects on postoperative survival, (2) the effects on tumor size and (3) immunological effects. The results obtained showed that immunopotentiators produced favorable effects. Cytokines should also be evaluated from the same standpoints as those for immunopotentiators. PMID- 2968071 TI - [Oral cytostatic agents in the prevention of urothelial carcinoma. Our experience]. PMID- 2968072 TI - Coronary flow characteristics of left coronary artery in aortic regurgitation before and after aortic valve replacement. AB - In 6 patients with pure aortic regurgitation, the velocity waveform in the left anterior descending coronary artery (LAD) was measured using an 80-channel 20-MHz pulse Doppler velocimeter before and immediately after aortic valve replacement. All patients showed normal coronary angiograms. Flow velocity was analyzed by both zero-cross and fast Fourier transform methods in real time. The LAD flow in severe aortic regurgitation was characterized by an increase in the systolic flow component, a small and rapidly decreased diastolic flow, an irregular velocity profile across the vessel, and a wider velocity spectrum. After aortic valve replacement, systolic flow decreased by 36.3 +/- 21.7% (p less than 0.01), whereas diastolic flow increased by 81.4 +/- 51.8% (p less than 0.05). The ratio of diastolic flow to total LAD flow increased from 63 +/- 13% to 82 +/- 7% (p less than 0.05). The velocity profile became more parabolic and had a narrow spectrum. These results suggest that operation for aortic regurgitation induces beneficial effects on the myocardial inflow immediately after valve replacement. PMID- 2968073 TI - Sickle cell heart disease. Two-dimensional echo and Doppler ultrasonographic findings in the hearts of adult patients with sickle cell anemia. AB - Sickle cell anemia (SCA) is a commonly diagnosed condition in the black population in the United States. With better medical management, patients with SCA are living longer and what were previously uncommon sequelae are being recognized frequently, including those involving the cardiovascular system. Previous reports on the cardiovascular effects of SCA have focused predominantly on children, but the conclusions remain controversial. To study this question further, 40 adult patients with documented SCA were examined using two dimensional and Doppler cardiac ultrasound. Valvular structures were normal despite an increased incidence of flow murmurs. Abnormalities were found more frequently in the left heart than the right, as manifested primarily by increased left ventricular mass, and left ventricular and left atrial dilatation with preservation of systolic function. Pulmonary hypertension, which was present in two thirds of the sample, was minimal to moderate as assessed by our Doppler technique. The effects of SCA on the heart seem to be minimal and similar to those of other anemias, predominantly confined to the left atrium and ventricle with passive elevation of pulmonary pressures. Clinical murmurs were most often physiologic; there was no association with myxomatous valvular degeneration or mitral valve prolapse. PMID- 2968074 TI - Significance of electrocardiographic isolated abnormal terminal P-wave force (left atrial abnormality). An echocardiographic and clinical correlation. AB - An abnormal terminal negative deflection in precordial lead V1 (PTFV1) is occasionally present as an isolated electrocardiographic finding. To determine the significance of this, 41 patients with PTFV1 greater than or equal to 0.04 mms were recalled for a repeated electrocardiogram as well as an echocardiographic examination. Patients were classified as cardiovascular normal (n = 15) or abnormal (n = 26). Left atrial enlargement was the most common echocardiographic abnormality found, but represented less than a third of the total. P-wave prolongation (greater than 110 ms) was present in 30 of the 41 patients and 21 of the 26 abnormal patients. This persisted into the second study, while the PTFV1 fell significantly in both the normal and abnormal groups. Among the P-wave abnormalities found, combinations were used to identify patients most likely to have cardiovascular disease. The most discriminating was an initial abnormal PTFV1 in combination with P-wave prolongation (sensitivity, 82%; specificity, 40%; positive predictive value, 70%; and negative predictive value, 55%). Maximal specificity (93%) and positive predictive value (88%) were achieved when P-wave prolongation and PTFV1 greater than or equal to 0.06 mms were present in both studies, although the sensitivity for this criterion was only 27%. The isolated P-wave abnormality described may be helpful in suggesting the presence of underlying cardiovascular disease and indicate the need for further evaluation. PMID- 2968075 TI - Diagnosis and course of nephrogenic ascites. AB - Nephrogenic ascites is a complex diagnostic problem with poorly understood pathophysiology. Morbidity and eventual mortality from this ongoing problem are significant. The diagnosis of nephrogenic ascites must be established by exclusion. We report the cases of nine patients investigated between 1978 and 1985. Laparoscopy, which was utilized in all nine patients, led to a specific diagnosis in two. We believe that a vigorous diagnostic evaluation, including laparoscopy, is essential in patients with chronic renal failure who develop persistent ascites. PMID- 2968076 TI - Cutaneous vasculitis and its relationship to systemic disease. PMID- 2968077 TI - Fixed drug eruption due to paracetamol. PMID- 2968078 TI - Contraception and the skin. PMID- 2968079 TI - Epicardial release of immunoreactive atrial natriuretic peptides in inside-out perfused rabbit atria. AB - An easy and convenient isolated atrial perfusion technique was developed. The effect of stretch of the atrial subpericardial myocytes was investigated in the inside-out perfused rabbit atria. Graded distension of the inverted atria was induced by changing the elevation of the atrial catheter tip. Intra-luminal volume expansion resulted in an increase in release of immunoreactive atrial natriuretic peptides (irANPs). The response was volume, or pressure dependent. Distension-induced release of irANPs occurred at the reduction of the distension. IrANPs in epicardial perfusate showed both high and low molecular weights. The major peak of irANP was observed at the corresponding fraction to the rat ANP-(1 28) in the Sephadex G-50 gel chromatography. The data suggest that the epicardial release of irANP is stretch-induced response and that the release may be involved in the regulation of cardiac function. PMID- 2968080 TI - Immunocytochemical localization of the Ca2+-ATPase polypeptide in human platelets. AB - Specific polyclonal antibodies raised against purified human platelet Ca2+-ATPase were used with protein A-gold immunocytochemistry to localize this protein in human platelets. Immunolabeling specifically detected Ca2+-ATPase over the surface connected membrane system (SCS) in sections of paraformaldehyde-fixed, Lowicryl-embedded platelets. The maximum density of label, determined by quantitative morphometric techniques, was observed over electron-dense regions within the SCS which may represent specialized structures for uptake and release of Ca2+. Less intense immunolabeling was observed over cytosol and may represent localization over the dense tubular system (DTS) which was not readily visualized under the processing procedures employed. PMID- 2968081 TI - Electroencephalographic effects of the new antidepressant paroxetine in the rabbit. AB - The electroencephalographic (EEG) effect of (-)-trans-4-(4'-fluorophenyl)-3 (3',4'-methylene-dioxyphenoxy-met hyl)piperidine hydrochloride (paroxetine, BRL 29060A) a new antidepressant, was investigated in conscious rabbits with chronic electrode implants and was compared with those of imipramine and amitriptyline. Paroxetine induced an arousal pattern of the spontaneous EEG consisting of low voltage fast waves in the cortex and synchronization of hippocampal theta waves with decreased amplitude, while imipramine and amitriptyline elicited drowsy patterns of the spontaneous EEG. Paroxetine failed to suppress the EEG arousal responses induced not only by auditory stimulation but also by electrical stimulation of the mesencephalic reticular formation, centromedian thalamus and posterior hypothalamus, whereas imipramine and amitriptyline markedly inhibited these responses. The EEG arousal response induced by i.v. injection of physostigmine 0.2 mg/kg was slightly enhanced by paroxetine, while the response was significantly suppressed by imipramine and amitriptyline. Paroxetine, imipramine and amitriptyline showed no significant effect on the photic driving response and recruiting response. Paroxetine did not show any effects on the limbic afterdischarges elicited by either hippocampal or amygdaloid stimulation, while imipramine and amitriptyline caused an initial suppression followed by slight enhancement of these afterdischarges. These results indicate paroxetine to be an antidepressant of a new type which induces a sustained arousal pattern of the spontaneous EEG and has no central anticholinergic action. PMID- 2968082 TI - Sinusoidal fetal heart rate pattern associated with gastroschisis. AB - A case of true sinusoidal fetal heart rate associated with fetal gastroschisis is presented. No other known etiologic factors associated with a sinusoidal pattern were present. The pathophysiologic mechanism in this case is unclear, but fetal anomalies may be associated with elevated arginine vasopressin levels leading to the distinctive sinusoidal fetal heart rate response. PMID- 2968083 TI - Program of the third annual World Congress on Biologically Active Atrial Peptides. Abstracts. PMID- 2968084 TI - Structure and function of human and murine receptors for IgG. PMID- 2968085 TI - The T cell response to the malaria circumsporozoite protein: an immunological approach to vaccine development. PMID- 2968086 TI - Prevention of cement dermatitis in construction workers with iron sulphate. PMID- 2968087 TI - Breast-feeding, dietary intake and weight-for-age of children in rural Burma. PMID- 2968088 TI - Attendance patterns for dental treatment of inoculation risk patients. PMID- 2968089 TI - Treatment of infectious and high-risk patients. PMID- 2968090 TI - Badges of the dental profession. British Society of Dentistry for the Handicapped. PMID- 2968091 TI - 'The dental health of the elderly mentally ill: the carers' perspective'. PMID- 2968092 TI - On the work of the radiologist--separation of image capture from image display. AB - A primary function of the radiologist is that of extracting sufficient information from images, in a finite time, so as to arrive at a patient management decision. Clinically important information must be present in the images and this information must be correctly perceived. This essay discusses the work of the diagnostician viewing prerecorded images. The potentials for enhancing this work by means of different image display techniques are examined. The application of stimulable phosphor computed radiographic techniques are discussed in detail as an example of a technology which clearly separates crucial portions of the imaging process. This separation of functions might improve general radiology in the near future. The principals of the discussion are not specific to any imaging modality or to any individual display technology. PMID- 2968093 TI - Digital radiography of the spine, large bones and joints using stimulable phosphor. Early clinical experience. AB - The value of digital radiography in musculoskeletal disorders was investigated by assessing its ability to depict anatomic structures and common radiologic features as compared with the conventional film-screen combination. The digital image that was frequency modified was superior to conventional films in delineating soft tissue structures and for areas with large attenuation differences. The conventional film-screen system was superior in depicting small anatomic structures and in identifying the zone close to prostheses. This was explained by the high spatial resolution of the conventional film system and the disturbing halo effect around the prosthesis seen with digital images. The halo effect is an overshoot caused by the unsharp masking operator, which was in this series not changed for individual examinations. The exposure (radiation dose) could be reduced to 50 per cent using the digital system, without any loss of information. PMID- 2968094 TI - Digital urography using stimulable phosphor. An experimental study in the rabbit. AB - In a urographic experimental study in six rabbits conventional film-screen images were compared with digitized images. Two digitized images were examined, one simulating the conventional film-screen combination, and one frequency modified. The nephrographic and excretory phases of four different doses of contrast medium (iohexol, 350 mg I/ml) were investigated in each rabbit (4, 2, 1, and 0.5 ml/kg body weight). No differences were found between the different doses of contrast medium for the two imaging systems. With digitized images the exposure (radiation dose) could be decreased to 12.5 per cent of the conventional film-screen combination without any loss of information. PMID- 2968095 TI - Digital radiography using laser stimulated luminescence for evaluation of urinary calcifications. AB - One hundred consecutive patients were investigated with both a conventional film screen combination and digital radiography using a laser stimulated luminescence technique in order to evaluate differences in detecting urinary tract stones. All patients were referred either for follow-up of previously known stones or for symptoms. The images were reviewed by two radiologists who had the single clinical information that the patients might have urinary stones. Following review, the two techniques were compared and a consensus was obtained. One hundred and eighteen stones were found in 41 patients without any statistical differences in sensitivity between the two imaging systems. PMID- 2968096 TI - Non-palpable breast lesions. Diagnostic criteria and preoperative localization. AB - Marking of non-palpable breast lesions for biopsy has become a routine procedure in patients in whom mammography has suggested malignancy. Between October 1981 and December 1985 a localization method was used in 123 patients. In 35 per cent of the biopsies a malignant lesion was disclosed. In patients with a mass only, malignancy was found in 37 per cent of the biopsies. If microcalcifications alone were the main reason for the biopsy, malignancy was disclosed in 25 per cent. If both a mass and microcalcifications were present, 52 per cent of the biopsies disclosed a malignant lesion. PMID- 2968097 TI - Large-image intensifier photofluorography and conventional radiography in pulmonary emphysema. Correlation with computed tomography. AB - Large-screen image intensifier (II) photofluorography was compared with full-size screen-film chest radiography in the diagnosis of pulmonary emphysema in 84 patients. Photospot films and conventional radiographs were interpreted independently by three radiologists. Computed tomography (CT) was used as an independent reference technique, and diagnostic performance of chest radiography in various CT patterns of emphysema was evaluated. The difference in diagnostic sensitivity for emphysema in favor of conventional chest radiography over photofluorography (0.65 versus 0.56) was statistically significant (p less than 0.05). Specificity of the imaging modalities was equal: 0.78 in full-size films and 0.77 in photospot films. All CT patterns of emphysema had great false negative response rates in chest radiography, which is an inaccurate technique for the diagnosis of emphysema. CT is required for reliable radiologic evaluation of emphysema. PMID- 2968098 TI - Increased arterial wall stiffness and thickness in medium-sized arteries in patients with insulin-dependent diabetes mellitus. AB - By means of ultrasonography, arterial wall stiffness, arterial wall thickness, and the elastic modulus of the common femoral artery were estimated in a group of 19 young insulin-dependent diabetics. The ultrasound technique for determination of these parameters is described as well as the echo-anatomy of the arterial wall. In accordance with a previous investigation a significant rise in arterial wall stiffness was found. Furthermore, there was a highly significant correlation between the stiffness and the thickness of the arterial wall. The elastic modulus also correlated to the stiffness. It is concluded that the diabetic macroangiopathy is characterized by an increased stiffness of the arterial wall caused by increased thickness as well as by progressive alterations of the elastic characteristics of the wall tissue. Possible pathogenetic reasons are discussed. PMID- 2968099 TI - Repeat femoral arteriography in hyperlipidemic patients. A study of progression and regression of atherosclerosis. AB - Femoral arteriography was performed in 62 patients with significant hyperlipidemia. Sixty were asymptomatic and two had intermittent claudication. The patients participated in a study aiming to demonstrate whether serum lipid lowering by drugs could influence the development of femoral artery atheromatous disease. Half of the patients were treated with fenofibrate and nicotinic acid and the other half served as a control group. At the first arteriography atherosclerotic lesions were found in 46 of the 62 patients (74%). Arteriography was repeated up to three times without complication. Visual analysis of angiograms revealed considerable inter-observer variation. An attempt was made to assess the angiograms by a computerized method which, however, still needs improvement and a computer designed for image analysis. Most patients had small or moderate atheromatous deposits in the femoral artery at the initial examination, in most cases showing no change during the study period of 18 months. Regression was found in five patients of the treated group, but in none of the control patients as judged by visual gradation (p less than 0.001). PMID- 2968100 TI - Quantitation of atherosclerosis in femoral arteriography with ECG gated exposures. AB - Using a densitometric method introduced in 1977 by Crawford et coll. the volumes of segments of the femoral artery were calculated from two angiographic series in each of 13 patients. ECG gated exposures were used to minimise the error of the method. The reproducibility of the method was found to be better than in a previous study in which ECG gating was not used. The method may therefore be of use in long term angiographic follow-up studies of atherosclerotic lesions, although the impact of different investigation parameters on methodologic precision is still not fully known. PMID- 2968101 TI - Significant reduction of the echogenicity of the compressed cavernous hemangioma. AB - Eleven hyperreflective hepatic nodules in eight patients were examined by means of intraoperative ultrasonography (US) with the transducer placed on the liver surface. The nodules included four cavernous hemangiomas, four hepatocellular carcinomas, one metastatic tumor, and two regenerative nodules in cirrhotic livers. When the relevant part of the liver was compressed during US observation, the echogenicity was reduced only in the case of cavernous hemangioma. This phenomenon is considered to be unique to cavernous hemangioma. PMID- 2968102 TI - Quantitative single photon emission computed tomography applied to a comparison of propanetetraphosphonate and albumin colloid for liver and spleen scintigraphy. AB - A clinical application of a previously developed technique for absolute quantitation of organ uptake of radioactivity is presented. The method, which is based on single photon emission computed tomography involving correction for scattering and attenuation of photons, enables an accurate in vivo determination of the amount of administered activity taken up in a specific organ. The technique was applied to a comparative clinical trial between propanetetraphosphonate and albumin colloid for liver and spleen scintigraphy. The mean uptake of the liver was 73 +/- 9 per cent using propanetetraphosphonate and 63 +/- 10 per cent using albumin colloid. The corresponding figures for the spleen were 6 +/- 3 and 9 +/- 4 per cent, respectively. The activity concentration of the different lobes of the liver, bone marrow and soft tissue was also estimated. Phantom studies showed that the total uptake in the liver could be determined with an accuracy of about 6 per cent. The accuracy of the clinical examinations was estimated to about 10 per cent. PMID- 2968103 TI - Transluminal angioplasty of a stenotic surgical splenorenal shunt. AB - A stenosis of a side-to-side splenorenal shunt was treated by percutaneous angioplasty two years after the performance of the shunt. After dilatation, there was a fall of the splenorenal pressure gradient from 28 to 17 cm H2O and good transanastomotic flow was re-established. As in other arterial and venous territories, angioplasty may be an interesting alternative to surgery. PMID- 2968104 TI - Radiographic findings in overlooked colon carcinomas. A retrospective analysis. AB - Double contrast barium enemas in 21 patients with primarily overlooked colon carcinomas were reviewed. The carcinomas were flat and measured less than 30 mm in 18 cases. These tumours constitute almost 1.4 per cent of the carcinomas diagnosed during a 12-year period. The retrospective analysis revealed the following signs of carcinoma: missing haustral folds, disharmony of interhaustral fold patterns; radiolucent filling defects; local contractions; residue-like masses and decrease of volume. In all cases, at least two secondary signs of carcinoma, range 1.7 to 3, were seen. PMID- 2968105 TI - Transanorectal ultrasonography in anal carcinoma. A prospective study of 21 patients. AB - Twenty-one consecutive patients with anal carcinoma of squamous cell type were evaluated by transanorectal ultrasonography (Bruel & Kjaeer) prior to radiation therapy. The normal anal anatomy, with three distinct layers, was easily demonstrated both in vitro and in vivo. The middle, low echogenic layer corresponded above the dentate line to the muscularis propria and more distally to the internal and external sphincters. A hypoechoic area, representing tumour, was detected in all patients. Using the ultrasound findings, it appeared possible to classify the depth of tumour invasion into four levels with respect to whether or not invasion had reached or penetrated beyond the muscular wall or into adjacent organs. Eighteen of 21 tumours had penetrated the muscular wall. In 3 cases low echogenic, rounded structures, interpreted as enlarged lymph nodes, were identified. The ultrasonographic findings were compared with digital staging. Tumour invasion had penetrated the muscular wall in 2 out of 3 stage T1 patients and in 10 out of 11 stage T2 patients. Prospective studies will show whether estimates of tumour size and depth of invasion in relation to various normal structures, as judged by ultrasonography, are of value prognostically and for the choice of therapy. PMID- 2968106 TI - Ultrasonography and computed tomography in pancreatic malignancy. AB - Previous reports have shown that ultrasonography (US) is inferior to computed tomography (CT) in detecting pancreatic carcinoma. Since our recent observations suggested the opposite we re-evaluated the accuracy of these two methods. Among 36 patients with malignant tumours the US was normal in one, in 4 it showed only secondary signs of tumour, and in 31 it revealed the tumour. The corresponding figures for CT were 2, 9 and 25, respectively. The conclusion is that US should be considered the primary method for imaging the pancreas. PMID- 2968107 TI - Roentgenologic assessment of spondylolisthesis. I. A study of measurement variations. AB - Variations when measuring slip and lumbar index were studied in three lumbar specimens with different degrees of artificially created L5-S1 spondylolisthesis. Lateral radiographs were obtained with the specimens tilted and/or rotated mimicking an every-day examination situation. Slip and lumbar index were measured; slip was estimated according to a measuring method advocated by BOXALL et coll. (3) but slightly modified by us. Six radiologists evaluated each film independently. The magnitude of interobserver and intraobserver errors and variations due to positioning was found to be equally large--about 15 per cent each. Although the total measurement variation was not equal to the sum of errors involved, radiologic evaluation of degree of spondylolisthesis was found to be highly unreliable. PMID- 2968108 TI - Computed tomography in low back pain before and after myelography. A qualitative comparison. AB - Forty-six patients with possible lumbar disk herniation were examined with computed tomography (CT) before and after lumbar myelography with metrizamide. Altogether 90 intervertebral spaces (L4/L5 and L5/S1) were analyzed with regard to the diagnostic information provided by pre- and postmyelography CT. Corresponding to the level of 73 intervertebral spaces (81%) the contrast medium filling of the dural sac was homogeneous, while corresponding to 17 spaces (19%) the filling was inhomogeneous, with layering of contrast medium. The demarcation of the disk margin and of the dural sac was poor at the level of 13 spaces (14%), the majority of which (9 spaces) belonged to the group with homogeneous dural sac filling. At the level of 4 intervertebral spaces the dural sac was isodense with the intervertebral disk. The overall diagnostic information from the premyelography examination was equal to the postmyelography examination at 66 intervertebral spaces (73%). The postmyelography CT examination showed the details better than the conventional CT at 11 intervertebral spaces (12%), and poorer than conventional CT at 13 intervertebral spaces (15%). It is concluded that in general a conventional CT examination is adequate in the evaluation of patients with suggested intervertebral disk herniation. The need for a postmyelography CT examination can be specified on the basis of the findings at myelography. PMID- 2968109 TI - Comparison of two- and three-dimensional methods for assessment of orientation of the total hip prosthesis. AB - A comparison of two- and three-dimensional methods for the determination of the orientation of total hip prostheses was made in a group of 57 patients. The acetabular inclination and the collum-diaphyseal angle measured on a.p. projections (2-D) were adequate in most cases for assessing how vertically the prosthetic component was inserted, but in individual cases with a high degree of version these measurements could be misleading. Anteversion measured in the transverse plane (2-D) was more sensitive to errors than planar anteversion measured as a rotation around the longest diameter of the ellipsoid projection of the acetabular opening, but it gave a rough estimate of the relation of the prosthetic components. Determination of the spatial (3-D) orientation of the components provides a precise estimate of the component relations. PMID- 2968110 TI - The percentage of migration as indicator of femoral head position. AB - In childhood subluxation of one or both hips may develop rather insidiously. For lack of generally accepted objective methods of assessment, ambiguous interpretations of findings in serial examinations are common. Many subluxations are overlooked during the early stages. In order to overcome such disadvantages, determination of the percentage of migration seems to be a reasonably easy and reliable technique facilitating evaluation of impending dislocation. This investigation was carried out in order to establish norms applicable to patients in the pediatric age interval. The 98th percentile of migration increases with age from 16 per cent in patients less than 4 years of age to 24 per cent in patients greater than or equal to 12 years. Higher figures represent subluxation. If the migration exceeds 80 per cent a manifest luxation is present. A difference in migration between the two hips larger than 12 per cent indicates abnormality calling for clinical and radiologic follow-up. PMID- 2968111 TI - Prediction of the stability of minimally displaced fractures of the lateral humeral condyle. AB - Fractures of the lateral humeral condyle run a great risk of subsequent displacement. This is also true--although to a lesser extent--for primarily non displaced or minimally displaced (less than or equal to 2 mm) fractures. The present retrospective investigation aimed at defining radiographic criteria in order to predict the stability of non-displaced or minimally displaced fractures in 159 children. The fractures were classified according to radiographic criteria into 3 groups considered to represent stable, ambiguous and unstable fractures. Additional displacement while immobilized in plaster occurred in 2.6 per cent of the fractures classified as stable and in 44.4 per cent of those classified as unstable (p less than 0.001); in the ambiguous group subsequent displacement occurred in 24.1 per cent. It is concluded that the radiographic criteria should be useful to predict the stability of non-displaced or minimally displaced fractures of the lateral humeral condyle in children. PMID- 2968112 TI - Contrast media induced ventricular fibrillation during coronary angiography in dogs. AB - This study is designed to determine which is more critical for contrast media induced ventricular fibrillation (VF), hyperosmolality or lack of sodium ions. The right coronary artery in the canine model was infused continuously with contrast media (CM) for 25 seconds or until ventricular fibrillation occurred. The test solutions included isotonic saline, 305 mg I/ml of meglumine diatrizoate, 370 mg I/ml of meglumine/Na diatrizoate and 370 mg I/ml of metrizamide. The incidence of VF with meglumine/Na diatrizoate was significantly higher than with metrizamide (p less than 0.05). These observations suggest that the hyperosmolality is more critical than the lack of sodium ions in CM when metrizamide is compared with conventional ionic CM. On the other hand, the results showing that contact time until VF was shorter with meglumine diatrizoate than with meglumine/Na diatrizoate may reflect the importance of sodium ions in CM with similar osmolality. PMID- 2968113 TI - A 'split-away technique' for percutaneous nephrostomy. AB - The currently used techniques for nephrostomy sometimes fail to advance the drainage catheter through the nephrostomy track into the renal pelvis. To solve this problem a new technique has been developed. The improvement with this technique is the dilatation of the track with polypropylene dilators and a split away sheath. The split-away sheath is threaded on the largest dilator. When the dilator is removed the drainage catheter is coaxially passed through the split away sheath into the renal pelvis. The sheath is then removed. The technique has been used successfully in 32 patients without complications. PMID- 2968114 TI - Patient exposure when using 99Tcm-DTPA for evaluation of cerebrospinal fluid shunt patency. AB - The absorbed dose from 99Tcm-DTPA was estimated from 12 investigations in 9 patients who were evaluated for the patency of ventriculo-peritoneal shunts. The distribution of 99Tcm-DTPA in the ventricles, urinary bladder and peritoneal cavity was determined in regions of interest from repeated static gamma camera images. The effective dose equivalent was calculated to be less than 0.10 mSv for an injected activity of 15 MBq. PMID- 2968115 TI - Coronary angioplasty in pregnancy. AB - Myocardial infarction is rare in pregnancy. A 30 year old white primigravida had an anterior infarct at 20 weeks' gestation, which was followed by troublesome angina. Coronary angiography showed a tight stenosis of the left anterior descending coronary artery. This was treated successfully by percutaneous transluminal coronary angioplasty. PMID- 2968116 TI - Low second trimester maternal serum unconjugated oestriol in pregnancies with Down's syndrome. AB - Second trimester maternal serum unconjugated oestriol levels were measured in the stored serum samples from 22 pregnancies associated with Down's syndrome and 110 unaffected control pregnancies, matched for maternal age, gestational age, duration of storage of the serum sample, smoking habits and maternal weight. The serum unconjugated oestriol level of each affected pregnancy was expressed as a multiple of the median (MoM) of its five matched controls. The unconjugated oestriol levels were significantly lower in the affected pregnancies than in the unaffected pregnancies; the median MoM was 0.79 (P less than 0.05). This association between low serum unconjugated oestriol and fetal Down's syndrome early in pregnancy raises the possibility that serum unconjugated oestriol measurement may be added to maternal age and alpha-fetoprotein measurement in the antenatal screening for Down's syndrome. PMID- 2968117 TI - Bacteriological contamination during laparoscopy with dye injection. AB - Peritoneal contamination with bacteria during the laparoscopy and dye procedure was studied in 38 subfertile women by culturing samples of peritoneal fluid, aspirated before, and after, injection of dye solution. Bacteria were carried with the dye into a previously sterile peritoneal cavity in 34 women (90%). These vaginal organisms were present in low concentrations, not exceeding 10(5) organisms/ml. These findings explain the low incidence of acute salpingitis after tubal patency tests, but the selective use of antibiotic prophylaxis is recommended. PMID- 2968118 TI - Diethyl pyrocarbonate reaction with the lactose repressor protein affects both inducer and DNA binding. AB - Modification of the lactose repressor protein of Escherichia coli with diethyl pyrocarbonate (DPC) results in decreased inducer binding as well as operator and nonspecific DNA binding. Spectrophotometric measurements indicated a maximum of three histidines per subunit was modified, and quantitation of lysine residues with trinitrobenzenesulfonate revealed the modification of one lysine residue. The loss of DNA binding, both operator and nonspecific, was correlated with histidine modification; removal of the carbethoxy groups from the histidines by hydroxylamine was accompanied by significant recovery of DNA binding function. The presence of inducing sugars during the DPC reaction had no effect on histidine modification or the loss of DNA binding activity. In contrast, inducer binding was not recovered upon reversal of the histidine modification. However, the presence of inducer during reaction protected lysine from reaction and also prevented the decrease in inducer binding; these results indicate that reaction of the lysine residue(s) may correlate to the loss of sugar binding activity. Since no difference in incorporation of radiolabeled carbethoxy was observed following reaction with diethyl pyrocarbonate in the presence or absence of inducer, the reagent appears to function as a catalyst in the modification of the lysine. The formation of an amide bond between the affected lysine and a nearby carboxylic acid moiety provides a possible mechanism for the activity loss. Reaction of the isolated NH2-terminal domain resulted in loss of DNA binding with modification of the single histidine at position 29. Results from the modification of core domain paralleled observations with intact repressor. PMID- 2968119 TI - Comparison of the active calcium extrusion, calcium buffering capacity and ATPase activity in rabbit reticulocytes and mature red cells. AB - The purpose of the present work was to study the changes in the pattern of calcium homeostasis following the loss of intracellular organella during red cell maturation. Reticulocytes and mature red cells were prepared from anaemic rabbits blood after daily bleeding. Experimental protocols were designed to study the calcium buffering capacity in intact and digitonin-disrupted cells, the calcium pumping rate and, the Ca2+-translocating ATPase activity in the aforementioned red cells subpopulations. In digitonin-disrupted cells, a vesicular calcium pool, sharing the properties of mitochondria, could be detected in reticulocytes but no in mature red cells. Calcium content and calcium buffering capacity were significantly lower in reticulocytes than in mature red cells. The pattern of active calcium extrusion was quite similar in the two cell subpopulations, although reticulocytes had somewhat higher calcium affinity. Besides, an estimation of the calcium pumping rate gave higher values in reticulocytes than in mature erythrocytes. These values were 21 and 9 mmol/l cells per h, respectively. Maximal activities of the high-affinity (Ca2+ + Mg2+)-ATPase and basal Mg2+-ATPase were significantly higher in reticulocytes than in mature red cells, but no differences were observed regarding calcium affinity. The results show that changes in the properties of the Ca2+-translocating ATPase and intracellular calcium buffering systems are mechanisms involved in the process of red cell maturation. PMID- 2968120 TI - The comparative influence of substituted phenols (especially chlorophenols) on yeast cells assayed by electro-rotation and other methods. AB - The toxicity of 31 phenols was studied by electro-rotation of yeast cells. Control yeast cells show both anti-field and co-field rotation, depending upon the field frequency applied. After treatment with supra-threshold amounts of phenols the anti-field rotation is weakened or abolished and a stronger co-field rotation can be seen. The proportion of cells showing the co-field rotation was found to be a sensitive measure of toxicity. Doses of 2.2 mumol/l of pentachlorophenol, or of 0.3 mumol/l of pentabromophenol were detectable after 3 h incubation at pH 4.0. At a given pH, the toxicity of the chlorophenols correlated extremely well with their octanol:water partition coefficients (Pow). The complete set of phenols showed fair overall correlation with Pow, but less good correlation with their acidity constants (pKa). In particular the toxicity of a given phenol was less than predicted from its pKa if the incubation pH was higher than the pKa. Biochemical assays on 23 of the phenols showed that the rotational sensitivity runs closely parallel to the sensitivities of cell growth rate and of the plasmamembrane ATPase, but less closely to the inhibition of purine incorporation. It appears that the electro-rotation method provides a useful and rapid test for the presence of organic ecotoxins. The test enables us to distinguish differences between single cells, and is comparable in sensitivity to biochemical tests that use vesicles or homogenates derived from a cell population. PMID- 2968121 TI - Stimulus-response coupling in marine sponge cell aggregation: lipid metabolism and the function of exogenously added arachidonic and docosahexaenoic acids. AB - Cells of the marine sponge, Microciona prolifera, the most ancient of the animal cells which clump on recognition, resemble neutrophils and platelets in undergoing stimulus-response coupling when exposed to Ca2+ ionophores and phorbol esters. We have studied lipid content and remodelling in sponge cells by thin layer, gas-liquid, and high-performance liquid chromatography (HPLC) analyses supplemented by ultraviolet and mass spectroscopy. Phosphatidylcholine (PC) (35.6%), phosphatidylethanolamine (PE) (27.4%) and phosphatidylserine (PS) (21.4%) constituted the bulk of phospholipids detected. The major fatty acids were all polyenoic; 22:6 (22%), 26:2 (17%) and 26:3 (15%). Arachidonic acid (20:4), present as 2.7% of total phospholipid, and docosahexanoic acid (22:6) were found to elicit aggregation of sponge cells when added (10 microM) in synergy with ionomycin (1 microM), resembling in their effects those of phorbol esters (but not phorbol) and 1-oleyl-2-acetylglycerol (OAG). Moreover, 20:4 and 22:6, as well as phorbol ester and OAG, overcame the block to aggregation imposed by colchicine and vinblastine. Kinetic studies of lipid remodelling showed that aggregating cells diverted [14C]22:6 or [14C]20:4 from triacylglycerol into diacylglycerol and phospholipids; appearance of label in phosphatidic acid and phosphatidylinositol (PI) anteceded labeling of phosphatidylcholine. In unstimulated cells, [14C]22:6 was rapidly incorporated into phosphatidylcholine with little accumulation in phosphatidate. Although 22:6 and 20:4 resembled OAG and phorbol esters in overcoming the effects of colchicine and vinblastine (which had no effects on overall lipid metabolism), they did not reverse the block to aggregation of nordihydroguaiaretic acid (NDGA) (which inhibited lipid metabolism). Under none of these circumstances was 22:6 or 20:4 converted to cyclooxygenase or lipoxygenase products in the course of aggregation: all labeled acyl groups remained present as unmodified fatty acids on alkaline hydrolysis. These data not only extend the observations of Muller et al. (J. Biol. Chem. 262 (1987) 9850-9858) on the role of phosphoinositides and C kinase in marine sponge cell aggregation, but also demonstrate that sponges form diacylglycerols in the process. We suggest that exogenous 22:6 and 20:4 (like phorbol esters or OAG) can substitute for endogenous diacylglycerol in the activation of protein kinase C. PMID- 2968122 TI - Regulatory aspects of mitochondrial phospholipase A2 from rat liver: effects of proteins, phospholipids and calcium ions. AB - This paper deals with the search for specific inhibitors or activators of the mitochondrial phospholipase A2. Convincing evidence for the existence of proteins in the mitochondrial or cytosolic fraction that function as specific regulators of this enzyme was not obtained. The enzymatic activity appeared to be inhibited at low substrate concentrations by lipocortin isolated from human monocytes. However, at higher substrate concentrations, the inhibition disappeared, suggesting either that lipocortin sequestered the phospholipid substrate or that the putative inactive complex of enzyme and lipocortin dissociated in the presence of excess phospholipids. The hydrolysis of the neutral phospholipid phosphatidylethanolamine was stimulated by the presence of cardiolipin and phosphatidylglycerol. It is unlikely that this is caused merely by the negative charge of these phospholipids, since other negatively charged phospholipids did not show this effect. Using a phospholipid extract from mitochondria as substrate, the enzymatic activity as a function of the Ca2+ concentration was determined. Only one enzyme activity plateau was observed. The calculated KCa2+ value of 0.05 mM suggests that the mitochondrial phospholipase A2 could be regulated strictly by the modulation of the free Ca2+ concentration in vivo. The two activity plateaus observed previously upon variation of the Ca2+ concentration using phosphatidylethanolamine as substrate could be explained by a Ca2+-induced transition of the phospholipid structure. PMID- 2968123 TI - [Contribution to studies on the treatment of cancer of the prostate with D-Trp6 LH-RH]. AB - The authors review 55 cases of adenocarcinoma of the prostate, from July 1982 to June 1983. The mean age of these patients was 61.5 years. None of them underwent orchiectomy. They were all treated with an LH-RH analogue: Decapeptyl. In 74.5% of the patients a consistent decrease of tumour mass in the prostate to about half its original size was observed, in association with a shrinking of the metastases. The Karnofsky index improved in 89% of the patients. The authors believe that Decapeptyl is preferable to surgical castration because it allows an improvement in pain symptoms, a regression of the primary tumour and metastases and achieves a better balance of serum levels of testosterone, FSH and LH. PMID- 2968124 TI - [Effects of a lymphocytic chalone-containing preparation on the mitotic activity of mouse thymocytes in vivo]. AB - It has been shown that chalone-containing substance obtained from the rat spleen decreased the mitotic activity of mouse thymocytes upon its intraperitoneal administration to animals. The maximum inhibition was observed 45 min after the injection. Thus, the substance examined has the property of G2 chalone. PMID- 2968125 TI - Activity of immobilized alpha-amylase. AB - 1. alpha-Amylase immobilized on polyamide 11 showed higher specific activity and retention of activity than the derivatives employing polyacrylamide and polyethyleneterephthalate as supports. 2. Polyamide 11 and polyethyleneterephthalate alpha-amylase derivatives exhibited a higher extent of multiple attacks on starch than the water-soluble enzyme whereas the polyacrylamide derivative presented less. 3. The polyamide 11 alpha-amylase derivative acted on amylose-azure in the same way as the water-soluble alpha amylase. PMID- 2968126 TI - The influence of castration and antiandrogenic treatment on the sensitivity of rat seminal vesicle to sympathomimetic drugs. AB - The sensitivity of seminal vesicles to sympathomimetic drugs and the plasma concentrations of testosterone and gonadotropins (LH and FSH) were examined in adult male albino rats divided into three groups: normal, castrated and Androcur treated. Dose-effect curves for adrenaline and noradrenaline were obtained in the presence and absence of cocaine or corticosterone and the parameter pD2 was determined. Castration and Androcur treatment induced a decrease in seminal vesicle sensitivity to drugs (shifting of dose-effect curves to the right, decreased pD2). Cocaine shifted the adrenaline and noradrenaline dose-effect curves to the left in three groups. Corticosterone did not modify the dose-effect curves of adrenaline and noradrenaline. Plasma testosterone levels were decreased by castration and Androcur-treatment although the LH and FSH concentrations were markedly altered by castration but were unaltered by the antiandrogenic treatment. The results suggest that the androgen may play an important role in the sensitivity of the seminal vesicle to sympathomimetic drugs through control of the neuronal uptake of catecholamines. PMID- 2968127 TI - Applicability of the equations of Freundlich and Langmuir to the adsorption of the azo dye procion scarlet on paramorphic colonies of Neurospora crassa. AB - Experiments on the adsorption of Procion Scarlet MX-G by normal hyphae and by paramorphic colonies of Neurospora crassa were performed at pH 2.5, 4.5 and 6.5 at 30 degrees C. The measured adsorption isotherms were evaluated by the Freundlich and Langmuir equations. The removal of dye was most effective at pH 2.5 and more dye was adsorbed per unit mass of cells in the paramorphic cultures than in the normal hyphae. The statistical tests showed Langmuir's equation to give a better fit to the adsorption data. PMID- 2968129 TI - Bilateral turn-table rectus abdominis myocutaneous flaps for an upper abdominal recurrent dermatofibrosarcoma. AB - The use of two "turn-table" rectus abdominis myocutaneous flaps is described using skin extensions from well above the costal margin, to close a large upper abdominal defect. The chest donor site was covered with a deltopectoral flap with primary closure, helped by postoperative arm positioning. PMID- 2968128 TI - Endometriosis. AB - Endometriosis occupies a unique position in medicine. The natural history of the disease is uncertain, its precise aetiology is unknown, the clinical presentation is inconsistent, and the treatment is poorly standardized. This review attempts to draw some conclusions from the growing number of publications devoted to the subject, and to provide guidelines for patient management. PMID- 2968130 TI - Response to D-Trp-6-luteinising hormone releasing hormone (Decapeptyl) microcapsules in advanced ovarian cancer. PMID- 2968131 TI - Vertebral osteomyelitis in the elderly. PMID- 2968132 TI - Provision of aids. PMID- 2968133 TI - Exfoliative dermatitis after amiodarone treatment. PMID- 2968134 TI - Excitatory effects of atrial natriuretic peptide on rat subfornical organ neurons in vitro. AB - Atrial natriuretic peptide (ANP) was applied in the solution bathing the rat subfornical organ in vitro and the response of 170 neurons recorded extracellularly. A significant change in firing rate was found in 20% of neurons tested at a concentration of 1 X 10(-10) M. At higher concentrations 45% of neurons responded. The response was usually a brief excitation. Tests of a range of concentrations on each of 6 neurons showed the maximal excitation was attained at 5 X 10(-9) M. The excitatory responses were generally (41/51) not affected by synaptic blockade indicating a direct action of ANP. The excitatory effect of ANP upon subfornical neurons can be contrasted with the generally inhibitory effect of changes in osmotic pressure. Neurons excited by ANP also responded to 5% osmotic pressure changes in the proportions (28/43) expected from the known response rate to osmotic pressure changes (66%). The commonest pattern of response (16/28) was excitation by hyper-osmotic solutions and inhibition by hypo osmotic solutions. PMID- 2968135 TI - Recognition of ill-defined signals in nucleic acid sequences. AB - A set of programs has been developed for the definition and handling of nucleic acid sequence consensus information. The sequences of known genetic control signals are combined in a matrix. The origins and positions of the signals are recorded. Old matrices can be updated dynamically: new signals are included and obsolete ones deleted. Matrices of several different types are computed optionally. Several of these matrices can be combined to find possible new signals. The use of matrices allows the exact quantification of signal qualities. The described programs are part of a program library named GENEXPERT. Application examples given are the search for tRNA genes and the search for promoters in the bacteriophage lambda genome. PMID- 2968136 TI - [Changes in blood pressure, heart rate and hormones after injection of ANF (atrial natriuretic factor) in patients with primary hyperaldosteronism]. PMID- 2968137 TI - Functional impairment induced by lipophilic cationic compounds on mitochondria. AB - Lipophilic cations, such as rhodamine 123, have selective anticarcinoma activity both in epithelial-derived tumor cells and in tumor cells injected into mice. The mechanism by which rhodamine 123 and safranin have their effect on mitochondrial function was examined. Rhodamine 123 and safranin inhibit the stimulation of mitochondrial respiration by ADP in a similar concentration range. This inhibition occurs whether the mitochondria are respiring on succinate as a substrate or on ascorbate plus tetramethylphenylenediamine. ATP hydrolysis was stimulated twofold by high lipophilic cation concentration. These results demonstrate that rhodamine 123 and safranin affect oxidative phosphorylation in a similar fashion. PMID- 2968138 TI - Atrial natriuretic factors and the brain. Satellite symposium of the first joint meeting of the International Society for Neurochemistry and the American Society for Neurochemistry. Margarita Island, Venezuela, June 6-8, 1987. Proceedings. PMID- 2968139 TI - Atrial natriuretic factors and the brain. PMID- 2968141 TI - Distribution of atrial natriuretic factor-like immunoreactivity in the brain of the frog Rana ridibunda. AB - The localization of atrial natriuretic factor (ANF)-like immunoreactivity in the central nervous system of the frog Rana ridibunda was examined by the indirect immunofluorescence technique, using an antiserum against synthetic ANF (Arg101 Tyr126). Immunoreactive cell bodies were principally found in the dorsal and medial pallium, the medial septal nucleus, the ventrolateral and anteroventral areas of the thalamus, the lateral forebrain bundle, the posterolateral thalamic nuclei, the preoptic nucleus, the dorsal infundibular nucleus, and the anteroventral tegmentum nucleus of the mesencephalon. Numerous cell bodies and a very dense fiber bundle were visualized in the interpeduncular nucleus. All the areas mentioned above contained a high density of immunoreactive fibers. In addition, the amygdala, the infundibular nucleus, the median eminence, and most of the areas of the mesencephalon contained a moderate number of ANF-positive nerve processes. In the frog pituitary, fibers and nerve terminals were found in the peripheral zone of the neural lobe. The intermediate and anterior lobes of the frog pituitary were totally devoid of ANF immunoreactivity. These results indicate that ANF-like material is widely distributed in the frog brain and that ANF may be involved in various brain functions including neuroendocrine regulations. PMID- 2968140 TI - Central interaction of the brain atrial natriuretic polypeptide (ANP) system and the brain renin-angiotensin system in ANP secretion from heart--evidence for possible brain-heart axis. AB - To elucidate the involvement of the brain renin-angiotensin system and the brain atrial natriuretic polypeptide (ANP) system in the regulation of ANP secretion from the heart, the effects of intracerebroventricular administration of angiotensin II and ANP on the plasma ANP level were examined in conscious unrestrained rats. The intracerebroventricular administration of angiotensin II at doses of 100 ng and 1 microgram significantly enhanced ANP secretion induced by volume-loading with 3-mL saline infusion (peak values of the plasma ANP level: control, 220 +/- 57 pg/mL; 100 ng angiotensin II, 1110 +/- 320 pg/mL, p less than 0.01; 1 microgram angiotensin II, 1055 +/- 60 pg/mL, p less than 0.01). The intracerebroventricular injection of angiotensin II at the same doses alone had no significant effect on the basal plasma ANP level. The enhancing effect of central angiotensin II on ANP secretion induced by volume-loading was significantly attenuated by pretreatment with the intravenous administration of the V1-receptor antagonist of vasopressin or with the intracerebroventricular administration of phentolamine. The intracerebroventricular administration of alpha-rANP(4-28) (5 micrograms) had no significant influence on the basal plasma ANP level; however, it significantly attenuated central angiotensin II potentiating effect of volume-loading induced ANP secretion. These results indicate that the brain renin-angiotensin system regulates ANP secretion via the stimulation of vasopressin secretion and (or) via the activation of the central alpha-adrenergic neural pathway, and that the brain ANP system interacts with the brain renin-angiotensin system in the central modulation of ANP secretion from the heart.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968142 TI - Bioactive atrial natriuretic factor-like peptides in rat anterior pituitary. AB - The presence of biologically active atrial natriuretic factor (ANF)-like peptides was demonstrated in rat anterior pituitary. ANF-like immunoreactivity was detected in rat anterior pituitary by specific radioimmunoassay and was extracted from rat anterior pituitary homogenates by heat-activated Vycor glass beads; extracts were purified by reverse-phase high performance liquid chromatography. Two peaks containing ANF immunoreactive material were obtained. The first peak was eluted from the C18 mu Bondapak column at a position similar to the 28-amino acid carboxy terminal peptide (Ser99-Tyr126)-ANF of prohormone. The second peak had the same pattern of elution as the 126-amino acid prohormone, (Asn1-Tyr126) ANF. The biological activity of the smaller molecular weight peptide (28 amino acid) was assessed by its inhibitory effect on 10(-8) M ACTH-stimulated aldosterone secretion in rat zona glomerulosa cell suspension. This ANF-like material also displaced I125-labelled ANF from rat glomerular receptors with a potency similar to synthetic (Arg101-Tyr126)-ANF. Immunocytochemical localization revealed a distribution of ANF-stained cells similar in pattern and location to that of gonadotrophs. These results suggest the existence of biologically active ANF-like peptides and ANF prohormone within the anterior pituitary. However, their role remains to be elucidated. PMID- 2968143 TI - Natriuretic and diuretic action of centrally administered rat atrial natriuretic peptide (99-126): possible involvement of aldosterone and the sympathoadrenal system. AB - Intracerebroventricular (ICV) administration of rat atrial natriuretic peptide (99-126) (rANP) to conscious male hydrated rats resulted in a dose-related increase in urinary volume and sodium excretion over a 6-h period of urine collection. A diminished mineralocorticoid effect on the kidneys may explain the natriuretic phenomenon. This hypothesis was tested by ICV rANP injection (1.25 microgram/5 microL) in conscious hydrated rats pretreated beforehand with d aldosterone (20 micrograms/kg, ip). Although the absolute amount of sodium excreted was reduced, aldosterone did not affect rANP-induced sodium output at 1 and 3 h. Rats that were sham-operated or bilaterally adrenalectomized after 4 days were pretreated with aldosterone and given an oral water load followed by ICV rANP or saline. The possible participation of the peripheral sympathetic nervous system in the central action of rANP was evaluated in rats pretreated with 6-hydroxydopamine. In sympathectomized and adrenalectomized rats natriuresis and diuresis were still evident after rANP. Our results indicate that the natriuretic effect of ICV rANP is independent of mineralocorticoids. Likewise, diuresis and natriuresis can occur in the absence of the adrenal glands and are independent from the neural tone that the adrenergic system exerts on sodium reabsorption. PMID- 2968144 TI - Further evidence for a hypothalamic site of action of atrial natriuretic factor: inhibition of prolactin secretion in the conscious rat. AB - The presence of atrial natriuretic factor (ANF) in the hypothalamus and pituitary gland suggests a possible neuroendocrine action of the peptide. Because ANF has been shown to alter the activity of hypothalamic neurons and to interact with brain dopamine systems, we examined the possibility that it might be involved in the hypothalamic control of prolactin (PRL) and thyroid-stimulating hormone (TSH) secretion. Neither basal not stimulated release of PRL or TSH from cultured dispersed anterior pituitary cells was altered by doses of ANF ranging from 10( 11) to 10(-6) M. Similarly, the in vitro inhibition of PRL release by dopamine was not affected by the presence of ANF (10(-7) M). Plasma levels of PRL and TSH in conscious male rats infused for 30 min with 0.01 or 0.1 microgram ANF-kg-1.min 1 did not differ significantly from those present in saline infused controls. Third-cerebroventricular injection of saline (2 microL) or saline plus ANF (0.02, 0.1, 1.0, or 2.0 nmol) did not significantly alter TSH secretion; however, injection of the two highest doses of ANF resulted in significant inhibition of PRL release. Levels of PRL remained significantly reduced for 90 min after injection of 2 nmol ANF. The results indicate that ANF can act centrally to alter the release of neural factors responsible for the hypothalamic control of lactotroph function. PMID- 2968146 TI - The income adequacy of provincial and territorial social assistance payments to non-institutionalized disabled adults: further evidence why Canada needs a guaranteed annual income policy. PMID- 2968145 TI - The effects of myoinositol on the autonomic neuropathy in the streptozotocin diabetic rat--a freeze fracture study. AB - This report describes a freeze fracture study of rat dorsal sympathetic chain in normal, streptozotocin diabetic on normal diet, and a group fed a 1% myoinositol normal diet. In nonmyelinated fibres, the diabetic group had significant loss of particles on the P face of juxta-axonal Schwann cell membranes, whilst myelinated nerves showed a profound loss of particles on P and E faces. In the group with adjuvant myoinositol the particle numbers were normal in both types of nerve. Axonal plasma membranes showed increased numbers of particles in the P face of the nonmyelinated membranes in the diabetic, but were normal in the myoinositol group. Myelinated axonal membranes showed no significant difference between the diabetic and the normal. Rats on a myoinositol diet showed a significant decrease in the E face particles. These results are consistent with those described in the sciatic myelinated nerves by earlier authors and reinforce the view that myoinositol is an important membrane constituent as phosphatidylinositol and is essential for the expression of normal protein particle numbers. PMID- 2968147 TI - Developing rural health services: an occupational therapy case study. PMID- 2968149 TI - Allergic skin eruption with desipramine treatment. PMID- 2968148 TI - Environmental control of the workplace. PMID- 2968151 TI - Chromosome breaks and fragile sites in leukemic bone marrow cells. AB - Bone marrow cells from leukemic and nonleukemic patients were examined for chromosome breakage in cultures treated with fluorodeoxyuridine (FUdR) and FUdR plus caffeine. The results indicate that the leukemic cells have more chromosome breakage than the nonleukemic cells when thymidylic synthetase is inhibited by FUdR. Addition of caffeine did not enhance this chromosome breakage. These findings of enhanced breakage by FUdR exposure in vitro, nevertheless, may suggest that leukemic cells in general are more susceptible to breakage than normal cells, thereby predisposing the former to secondary chromosome rearrangements. PMID- 2968150 TI - Anaplastic thyroid carcinoma. A clinical, histologic, and immunohistochemical study. AB - Twenty-six cases of anaplastic thyroid tumor were investigated and reclassified using immunoperoxidase techniques. Sections of the neoplasms were stained immunohistologically for the following thyroid associated antigens: (1) thyroglobulin, which shows a positive reaction with follicular cells of the thyroid; (2) calcitonin, which is positive in medullary carcinoma of the thyroid; and (3) leucocyte common antigen (LC), which identifies lymphomata and Factor VIII-related antigen for hemangioendothelioma. Using these methods, five cases were reclassified. Three cases were identified as lymphomata, one case was reclassified as medullary carcinoma of the thyroid, and one case was identified as hemangioendothelioma. Eleven cases were confirmed to be anaplastic carcinoma of the thyroid and ten cases were negative for all the antigens tested. There was a significant difference in the survival of the groups of patients mentioned above. Prognostic data support the suggestion that immunohistochemical methods should be used for the precise classification of anaplastic carcinoma. In this way, tumors such as malignant lymphoma and medullary carcinoma, which resemble anaplastic carcinoma histologically but have a better prognosis, can be identified. This is important for planning surgical procedures and choosing chemotherapy and/or radiotherapy. PMID- 2968152 TI - The linkage of health status changes and disability. AB - Forecasting health and functional status changes in elderly populations is a difficult task because forecasting must describe the linkage of chronic morbidity, and its progression, with different levels and types of disability. In this paper we present a model which links morbidity and disability and, with the use of multiple survey and epidemiological data sources, provide estimates of the long-term reduction in chronic disability that would result from changing the prevalence of specific chronic diseases by the control of major risk factors. The results show that many of the diseases which are currently the principal target of primary prevention have more impact upon mortality, and overall life expectancy, than they do on the age at onset of chronic disability. Thus, in order to increase the average number of years that elderly persons can expect to live with less disability, or less severe forms of disability, new prevention strategies and techniques may have to be developed. PMID- 2968153 TI - Unusually long inflation times during percutaneous transluminal coronary angioplasty. AB - Unusually prolonged balloon inflations (PBI) during percutaneous transluminal coronary angioplasty (PTCA) have been utilized in 26 consecutive patients to establish adequate coronary perfusion after shorter inflation times resulted in severe residual stenosis (N = 15), early arterial closure (N = 5), or extensive dissection (N = 6). Inflations lasted 1.5 to 15 min and 23/27 stenoses were successfully dilated. The average reduction in stenosis was 67% (90%-33%). Chest pain occurred in 34% vs. 7% (P less than 0.01), and ST changes occurred during 66% vs. 57% (NS) of PBI vs. short inflations. Four patients required coronary artery bypass surgery in spite of PBI, but none suffered an infarction. Electrocardiograms were unchanged in 22 patients and showed nonspecific ST-T wave changes in four. Cardiac enzymes obtained in nine patients failed to reveal a significant increase in serum concentration. We conclude that PBI is well tolerated and can successfully dilate lesions not responding to short inflations. PMID- 2968154 TI - Percutaneous femoral transluminal angioplasty of a right brachial artery occluded after Sones coronary angiography. AB - A case of percutaneous transluminal angioplasty (PTA) from the groin to a 7-cm right brachial artery occlusion in a 41-year-old man, having previously undergone Sones coronary angiography 2 months earlier, is presented. The post-PTA angiogram shows near-normal reconstitution of the total obstruction, persisting after 6 months with complete loss of collateral circulation. In a case of total obstruction of the brachial artery, femoral PTA should be considered before resorting to surgical treatment. PMID- 2968155 TI - Clinical experience with the Monorail balloon catheter for coronary angioplasty. AB - The Monorail balloon catheter is distinctly different from other current balloon catheters: the guidewire passes through the balloon itself, exits the catheter proximal to the balloon, and runs alongside its small shaft (3 French) through the guiding catheter. Monorail coronary angioplasty was attempted in 61 patients on 73 lesions with balloons from 2.0 to 3.7 mm. Angiographic success was obtained in 66 lesions (90%). For 15 lesions, balloon exchanges were needed. In three lesions, the Monorail balloon failed to cross the lesion, while a standard balloon succeeded; two lesions could not be crossed with any balloon. Vessel occlusion occurred in four patients: two had emergency surgery without infarct (one died suddenly 4 days later and one had a stroke 1 day later), one was recanalized with a standard balloon, and one had a myocardial infarct. Continuous infusion of urokinase was used until patient 3 in whom problems with the delivery system led to cardiocerebral air embolization (with complete recovery). No thrombotic complications were observed in the subsequent 58 patients with only a bolus of 10,000 U of heparin. The Monorail balloon facilitates contrast injections and balloon exchanges but appears more difficult to pass through tight lesions. Omission of the previously recommended infusion with a thrombolytic agent proved safe. PMID- 2968156 TI - Use of angioplasty guidewire for technically difficult angiography. AB - A new technique for negotiating severe vessel tortuosity or stenosis is described whereby a PTCA guidewire is initially used to cross the tortuosity or stenosed area. After introduction of a long-tipped Sones catheter over the PTCA wire and past the stenosis, the latter is exchanged for the usual 0.035-in guidewire. The Sones catheter is then removed and angiography proceeds as usual. PMID- 2968157 TI - Activation of transcription factor IIIC by the adenovirus E1A protein. AB - The factor(s) responsible for the adenovirus E1A-stimulated transcription of RNA polymerase III genes was localized previously in a chromatographic fraction containing transcription factor IIIC (TFIIIC). In further studies, two distinct forms of TFIIIC, which were chromatographically separable, generated VA gene protein complexes that were distinguished by gel shift assays. The form of TFIIIC that generated the more slowly migrating promoter complex had greater transcriptional activity in vitro, associated more rapidly with the promoter, and formed a more salt-resistant complex. Greater amounts of this more active form of TFIIIC resulted from either E1A expression during infection or growth of the cells in a higher concentration of serum, whereas template commitment assays indicated that overall TFIIIC concentrations remained unchanged during viral infection. The in vitro interconversion of the two forms of TFIIIC by phosphatase treatment suggests that transcriptional activation of RNA polymerase III genes can be mediated by phosphorylation of TFIIIC. PMID- 2968158 TI - An adenovirus E1A protein domain activates transcription in vivo and in vitro in the absence of protein synthesis. AB - We have shown previously that a synthetic peptide of 49 amino acids, encoding mainly adenovirus E1A protein domain 3 (PD3), functions as an autonomous transcriptional activator. Here we provide two lines of evidence showing that E1A transactivation does not require the induction of cellular protein synthesis. First, PD3 rapidly transactivates E1A-inducible early viral genes in the presence of inhibitors of protein synthesis, as demonstrated by microinjection-in situ hybridization experiments. Second, PD3 greatly stimulates transcription of E1A inducible genes in vitro. Mutant PD3 peptides with single amino acid substitutions in conserved cysteine residues are defective in transactivation both in vivo and in vitro. Our findings provide compelling evidence that protein synthesis is not required for E1A transactivation, and support a model in which E1A modifies the activity of a preexisting cellular protein(s) involved in the regulation of transcription. PMID- 2968159 TI - Purification and visualization of native spliceosomes. AB - Mammalian spliceosomes were purified in preparative amounts by gel filtration chromatography and shown to be functional by in vitro complementation experiments. The column fractions containing spliceosomes are enriched in the snRNAs U1, U2, U4, U5, and U6 and a subset of proteins present in the nuclear extract. Splicing intermediates, the entire set of snRNAs, and the enriched proteins can be immunoprecipitated with three different monoclonal antibodies that recognize snRNP determinants. At least one U1 snRNP is present in each spliceosome since the particles are quantitatively immunoprecipitated by an anti U1 snRNP monoclonal antibody. Examination of the spliceosome fractions by EM revealed a relatively homogeneous population of 40-60 nm particles with a striking morphology. Evidence that these particles are spliceosomes is their sensitivity to micrococcal nuclease, their ATP-dependent assembly, and their immunoprecipitation with a trimethyl cap monoclonal antibody. In addition, pre mRNA was visualized in the particles by EM. PMID- 2968160 TI - [An experimental investigation of the colored-IGSS method]. PMID- 2968161 TI - [Knowledge concerning cultivation of arterial smooth muscle cells]. PMID- 2968162 TI - [GFAP immunohistochemical analysis of Alzheimer type II cells in Wilson disease]. PMID- 2968163 TI - [Histological and histochemical changes in ovarian mucinous tumors. A study of 112 cases]. PMID- 2968164 TI - [The correlation between diseases and age (an analysis of 4584 autopsied cases)]. PMID- 2968165 TI - [Antigen-specific detection of circulating immune complexes in sera from patients with lung cancer]. PMID- 2968166 TI - [Ultrastructural study on the tracheal epithelium of hamsters induced by chemical carcinogen]. PMID- 2968167 TI - [Induction of lung tumors in mice by low doses of ionizing radiation and diethylnitrosamine]. PMID- 2968169 TI - [Histological study of experimental bladder tumors]. PMID- 2968168 TI - [A comparative study between ultrastructural changes of the photodynamic effect of photocarcinorin in killing human liver cancer cells]. PMID- 2968171 TI - [Immunohistochemical study of 59 cases of pituitary adenoma]. PMID- 2968170 TI - [Radiation damage of growing bone in rabbits]. PMID- 2968172 TI - [Ultrastructural study of neonatal lungs with respiratory distress syndrome]. PMID- 2968173 TI - [Penicilliosis marneffei]. PMID- 2968174 TI - [Retrospective analysis of infertile patients in a female reproductive endocrinologic clinic]. PMID- 2968175 TI - [Inhibitory effect of ligustrazine on acute and chronic pulmonary hypertension in rats]. PMID- 2968176 TI - [Dilative effect of injection of radix Angelicae, Salvia miltiorrhiza and ligustrazin on the pulmonary vessels in rats]. PMID- 2968177 TI - [Inhibitory effect of poriatin on murine leukemia 1210 cells]. PMID- 2968178 TI - [Immunologic studies on hemoglobin Bart's. IV. Application of anti-human Hb Bart's antibody for diagnosis of alpha-thalassemia traits]. PMID- 2968179 TI - [Sequence analysis of a beta-thalassemia gene from South China]. PMID- 2968180 TI - [Determination of sodium valproate in plasma by GLC]. PMID- 2968182 TI - [Relationship between gossypol concentration and testicular damage]. PMID- 2968181 TI - [Studies on malignancy regulation by heterologous hybridization between reticulocytes and a human plasmocytoma cell line (HMY-2)]. PMID- 2968183 TI - [Human natural infection by Echinochasmus perfoliatus in Anhui Province]. PMID- 2968184 TI - [Method of chemical determination of fusarin C]. PMID- 2968185 TI - Effects of anesthetic induction on myocardial function and metabolism: a comparison of fentanyl, sufentanil and alfentanil. AB - Anaesthetic induction may induce myocardial ischaemia. A prospective randomized trial was instituted to compare the effect on ventricular function and myocardial metabolism of induction with fentanyl (FEN) or its analogues sufentanil (SUF) or alfentanil (ALF) in 96 patients undergoing elective coronary artery bypass grafting (CABG). Haemodynamic, metabolic (coronary sinus oxygen and lactate extraction) and gated ventriculographic measurements were made awake pre induction (PRE), after induction (IND) and after intubation (INT). Induction was performed with FEN 75 micrograms.kg-1, SUF 15 micrograms.kg-1 or ALF 125 micrograms.kg-1 and metocurine. Fentanyl induction was associated with the greatest stability of mean arterial pressure (MAP), cardiac performance, and systolic function without associated myocardial lactate production. SUF produced the greatest depression of systolic function (p less than 0.05) but without haemodynamic instability or myocardial lactate production in all but one patient. Induction with ALF produced the greatest reduction in MAP (p less than 0.05) associated with the greatest decrease in diastolic compliance (p less than 0.05) and 50 per cent incidence of myocardial lactate production (p less than 0.05) with no significant change in coronary blood flow or myocardial oxygen consumption. PMID- 2968186 TI - Pharmacokinetics of sufentanil in patients undergoing renal transplantation. AB - Renal failure and chronic haemodialysis are often associated with alterations in fluid status and plasma proteins. These changes, in turn, may result in pharmacokinetic alterations in affected patients. The purpose of this study was to investigate the pharmacokinetics of sufentanil in chronic renal failure patients undergoing kidney transplantation. Ten male patients were studied. Following induction of anaesthesia each patient received sufentanil 2.0 micrograms.kg-1 IV with subsequent serial plasma sampling for drug measurement from one to 360 minutes. A biexponential equation provided the best fit of the sufentanil concentration data with mean +/- SEM distribution (alpha) and elimination (beta) half-lives of 2.9 +/- 1.3 and 176 +/- 87 minutes, respectively. The mean Vc and Vd beta values were 0.15 +/- 0.05 L.kg-1 and 0.85 +/- 0.16 L.kg-1, respectively; plasma drug clearance was 11.5 +/- 3.7 ml.kg-1.min 1. Mean values for K10, K12 and K21 were 0.15 +/- 0.06.min-1, 0.4 +/- 0.14.min-1 and 0.1 +/- 0.04.min-1, respectively. With the exception of Vd beta, these pharmacokinetic values are similar to those reported in previous studies in general surgical, elderly and burn patients. The Vd beta values observed in this study may have resulted from alterations in drug distribution or elimination following revascularization of the implanted kidneys. Nevertheless, it appears that modification of sufentanil doses is unnecessary in chronic renal failure patients undergoing renal transplantation. PMID- 2968187 TI - The mutagenicity and DNA base sequence changes induced by 1-nitroso- and 1 nitropyrene in the cI gene of lambda prophage. AB - The mutagenicity of 1-nitropyrene and its reduced metabolite 1-nitrosopyrene was determined in the lambda cI gene of an Escherichia coli uvr- lysogen. 1 Nitropyrene induced a mutation frequency of 3.8 x 10(-6), which was approximately 2-fold higher than the background mutation frequency, whereas an equimolar dose of 1-nitrosopyrene induced a much higher mutation frequency of 1.4 x 10(-4). Previous studies have established that both compounds form the same premutagenic lesion, viz. N-(deoxyguanosin-8-yl)-1-aminopyrene in bacterial DNA. In order to determine how this initial premutational lesion is converted to a stable heritable mutation, DNA sequences were determined for 30 mutations induced by 1 nitrosopyrene that mapped between bp 1 and 352 in the lambda cI gene of E.coli lysogens. We show here that these mutations are mainly frameshifts involving the addition or deletion of a single GC or CG base pair. A small proportion of mutations were base substitutions which were equally divided between transitions and transversions. These also occurred primarily at GC or CG sites. PMID- 2968188 TI - Early lesions induced by DHPN in Syrian golden hamsters: influence of concomitant Opisthorchis infestation, dehydroepiandrosterone or butylated hydroxyanisole administration. AB - The effects of concomitant Opisthorchis infestation and dehydroepiandrosterone (DHEA) or butylated hydroxyanisole (BHA) administration on dihydroxy-di-n propylnitrosamine (DHPN) induction of preneoplastic lesions were investigated in Syrian golden hamsters. Whereas parasite infection was primarily associated with first-order ductular proliferation in the liver and a secondary appearance of cholangiofibrotic lesions, DHEA treatment brought about increased carcinogen toxicity and enhanced generation of glutathione-S-transferase P (GST-P)-positive hepatocellular foci, liver cysts and focal proliferative changes in the pancreas. BHA also exerted an enhancing influence on pancreatic but not liver carcinogenesis. The results suggest that whereas alteration of DHPN metabolism by DHEA and BHA treatment effected changes at the initiation level, opisthorchiasis principally exerted an enhancing influence subsequent to carcinogen withdrawal. PMID- 2968189 TI - Phenotypic modulation of hepatocarcinogenesis and reduction in N nitrosomorpholine-induced hemangiosarcoma and adrenal lesion development in Sprague-Dawley rats by dehydroepiandrosterone. AB - Dietary administration of 0.25% dehydroepiandrosterone (DHEA) during and subsequent to 7 weeks treatment with N-nitrosomorpholine (NNM) resulted in significantly reduced development of adrenal cortical lesions and hemangiosarcomas in the liver. In addition, distinct phenotypic modulation of hepatocellular tumours was observed after combined hormone and carcinogen treatment. Thus the neoplasms were characterized by a higher degree of differentiation, lower mitotic rate and reduced potential for metastasis as compared to tumours observed after NNM alone. The data clearly indicate that DHEA exerts an inhibitory effect on both adrenal and hepatocarcinogenesis similar to that earlier reported for neoplasia in lung, thyroid, colon and skin. PMID- 2968190 TI - Computer microtechnology for a severely disabled preschool child. AB - The developments and considerations essential to providing microtechnological aids for a preschool severely disabled child confined to a bed in a hospital ward are described. This single case report illustrates the needs of a quadriplegic boy dependent on a ventilator via a tracheostomy. Provision of a computer, a variety of specially designed switches and software together with a self-driven powered wheelchair maximized expression of his developmental needs. The decision making process of providing the equipment is described as well as the problems encountered. His development in language and cognitive skills is discussed in the context of his change in experience and competence as he learned to control the technological aids. PMID- 2968191 TI - Monensin turns on microtubule-associated translocation of secretory granules in cultured rat atrial myocytes. AB - We have studied the effect of monensin on microtubule-associated translocation of atrial secretory granules in 5-7-day-old cultures of atrial myocytes from adult rats. Atrial granules and microtubules were localized by immunofluorescent microscopy of myocytes double-labeled with primary antibodies against atrial natriuretic polypeptide (ANP) and alpha-tubulin. In control myocytes, fluorescence due to atrial granules was predominantly localized to the perinuclear region containing the Golgi complex. After exposure for 30 minutes to monensin (0.5-5.0 microM), myocytes transiently contained conspicuous linear arrays of atrial granules associated with cytoplasmic microtubules. Thereafter, ANP fluorescence accumulated in subsarcolemmal foci at the cell periphery, while perinuclear ANP fluorescence faded. The monensin-induced redistribution of atrial granules was observable in both serum-containing and serum-free media and was unaffected by reducing external Ca2+ to low values, by inhibition of sarcoplasmic reticulum Ca2+ release with ryanodine, or by both. The redistribution was prevented by pretreatment with nocodazole, which fragmented microtubules and scattered Golgi complexes and the associated atrial granules throughout the cytoplasm. Radioimmunoassay showed that monensin seemingly decreased the rate of ANP secretion into the medium from 0.15 to 0.11 fmol/(hr.myocyte). These results suggest that monensin turns on microtubule-associated translocation of atrial granules from the perinuclear areas to the cell periphery by modifying the interaction between microtubules and atrial granules. Monensin also promotes movement of atrial granules along the microtubules but does not accelerate the release of ANP. PMID- 2968192 TI - Atriopeptin III induces early relaxation of isolated mammalian papillary muscle. AB - Atrial natriuretic peptide, released by mammalian atria in response to volume overload, induces vasodilation and natriuresis. In this study, a direct effect on cardiac mechanical performance was demonstrated. Atriopeptin III (10(-9)-10(-7) M) induced early relaxation and decreased peak twitch of isometric and isotonic twitches of isolated papillary muscles of cat and rat, without affecting maximal unloaded velocity of shortening. This effect resembled the effects of dibutyryl cyclic GMP and of sodium nitroprusside on cardiac muscle. The action of atriopeptin III, but not of dibutyryl cyclic GMP or sodium nitroprusside, was abolished by mechanically or chemically damaging the endocardial endothelial surface. Thus, the early relaxation of cardiac muscle induced by atrial natriuretic peptide may be mediated through receptors on the endocardial endothelium. PMID- 2968194 TI - Reversible induction of right ventricular atriopeptin synthesis in hypertrophy due to hypoxia. AB - Right ventricular hypertrophy produced in rats exposed to 10% oxygen for 3 weeks resulted in a ninefold increase in atriopeptin immunoreactivity (APir) and a 160 fold increase in atriopeptin messenger RNA (AP mRNA) in the right ventricular myocardium. A small but significant increase in left ventricular APir and AP mRNA was also present, probably representing the interventricular septum. Right atrial APir was decreased by 50%, but left atrial APir was not different from normoxic controls. Purification of ventricular tissue extracts by high-performance liquid chromatography revealed primarily the high molecular weight prohormone. The development of right ventricular hypertrophy and right ventricular APir content followed a similar time course, each evident at 7 days of hypoxia and reaching a plateau at 14 days. Hypoxia followed by normoxia caused right ventricular APir to fall to control levels within 3 days, despite persistent right ventricular hypertrophy. This data demonstrates that hypoxia can reversibly induce extra atrial expression of atriopeptin synthesis in the cardiac ventricle. PMID- 2968193 TI - Effects of chronic dobutamine administration on hearts of normal and hypertensive rats. AB - We have previously shown that physical conditioning in the rat improves cardiac mechanics and biochemistry and normalizes the cardiac contractile protein abnormalities associated with renovascular hypertension. Since chronic adrenergic stimulation with dobutamine simulates some aspects of physical conditioning, this study was undertaken to investigate the effects of chronic dobutamine administration on normal and hypertensive rat hearts. Four groups of female animals were studied: controls, dobutamine-treated (2 mg/kg twice daily), renovascular hypertensives, and dobutamine-treated hypertensives. Animals were killed after 8-10 weeks and cardiac histology, myosin biochemistry, and mechanics in an isolated heart perfusion apparatus were studied. Dobutamine, unlike hypertension, was not associated with histological evidence of myocardial damage but did increase cardiac mass by 10% and calcium-activated myosin ATPase activity by 13%. Hypertension was associated with a 24% increase in mass, a 24% decrease in ATPase activity, and a shift in the myosin isoenzyme pattern from V1 to V3. The combined stimuli caused additive hypertrophy (44%) and normalized myosin biochemistry and isomyosin distribution. Dobutamine treatment was not associated with significant improvements in pump or muscle function in control or hypertensive hearts. Thus chronic dobutamine treatment, like physical conditioning, induces a physiological cardiac hypertrophy in rats that is associated with improved myosin enzymology and normalization of the contractile protein abnormalities associated with hypertension. Unlike physical conditioning, however, these biochemical alterations do not result in improved contractile function as measured in an isolated buffer-perfused heart apparatus. PMID- 2968195 TI - Acceleration of the wavefront of myocardial necrosis by chronic hypertension and left ventricular hypertrophy in dogs. AB - Previous studies have shown that hypertension and left ventricular hypertrophy (HT-LVH) increase completed infarct size. Myocardial infarction progresses in a wavefront of myocardial necrosis from the subendocardium to the subepicardium. We tested two hypotheses: First, HT-LVH accelerates the wavefront of myocardial necrosis when compared with normotensive animals; and second, lowering of arterial pressure by infusing nitroprusside 1 hour after coronary artery occlusion exerts a salutary effect on infarct size. To test these hypotheses, systemic hypertension (mean aortic pressure = 141 +/- 3 mm Hg) and left ventricular hypertrophy (18% increase in left ventricular mass) were induced in dogs using a single-kidney, single-clip model. Seventeen adult mongrel dogs were used as controls. We measured mean aortic pressure, heart rate, left atrial pressure, and myocardial perfusion (microspheres) in several groups of normal and HT-LVH awake dogs. In two groups (normal and HT-LVH), 1 hour of circumflex coronary artery occlusion was followed by 4 hours of reperfusion. In two additional groups (normal and HT-LVH), 3 hours of circumflex coronary artery occlusion was followed by 90 minutes of reperfusion. In another group with HT LVH, nitroprusside was infused to reduce mean arterial pressure to 100 mm Hg beginning 1 hour after occlusion and was continued for the duration of reperfusion period (HT-LVH + N). Infarct size was assessed using triphenyltetrazolium chloride stain and risk area was determined using postmortem barium angiography. Fifteen of 17 (88%) control animals survived coronary artery occlusion, whereas only 17 of 42 (40%) dogs with HT-LVH survived coronary occlusion (p less than 0.05). Infarct-to-risk ratios in the various layers of the left ventricular wall were determined for survivors in all groups. After 1 hour of coronary occlusion more than twice as much mid-wall and epicardium was infarcted in the HT-LVH group compared with the control group. After 3 hours of coronary occlusion significantly more endocardium, mid-wall, and epicardium was infarcted in the dogs with HT-LVH. In the nitroprusside-treated HT-LVH dogs, the infarct sizes were similar to control animals. From these data we conclude: 1) the rate of infarction is accelerated in animals with HT-LVH; 2) nitroprusside infused 1 hour after coronary artery occlusion and continued throughout the reperfusion period exerts beneficial effect on infarct size when compared with control animals; and 3) acute coronary artery occlusion in animals with HT-LVH is associated with significantly greater mortality when compared with control animals. PMID- 2968196 TI - Assessment of immediate and long-term functional results of percutaneous transluminal coronary angioplasty. AB - Assessment of the functional significance of coronary artery lesions during cardiac catheterization has recently become possible by calculating coronary flow reserve from both myocardial contrast appearance time and density in the resting and hyperemic states determined from digitized coronary cineangiograms. However, the interobserver and intraobserver variabilities, as well as the short-, medium , and long-term variabilities of the coronary flow reserve measurements, have to be established before this technique becomes an acceptable means of assessing the immediate and long-term functional results of revascularization procedures such as percutaneous transluminal coronary angioplasty (PTCA). Variability was defined as the mean difference and standard deviation of the difference between duplicate determinations of coronary flow reserve. The intraobserver variability (mean difference +/- SD) in the measurement of coronary flow reserve was -0.01 +/- 0.07. Interobserver variability by two observers was +0.08 +/- 0.52. Short-term variability based on the analysis of two coronary cineangiograms taken 5 minutes apart was -0.02 +/- 0.26. Medium-term variability (coronary cineangiographies repeated 1-3 hours apart) was found to be -0.06 +/- 0.52. Long-term variability (coronary cineangiographies repeated 3-5 months apart) was 0.11 +/- 0.63. Having established the reproducibility of this radiographic method, we studied the prospective changes in coronary flow reserve in 25 patients undergoing PTCA for single vessel coronary artery disease. Coronary flow reserve measurements and quantitative coronary cineangiography were performed before, immediately after, and 3-5 months after PTCA. PTCA resulted in an immediate increase in coronary flow reserve from 1 +/- 0.3 to 2.3 +/- 0.6 with a concomitant increase in obstruction area from 0.9 +/- 0.3 to 3.3 +/- 0.7 mm2. Nine of the 25 patients developed restenosis defined as a diameter stenosis greater than 50% at follow up. The other 16 patients had a coronary flow reserve of 3.3 +/- 0.6, which was measured 3-5 months after PTCA. Coronary flow reserve measurement from digitized coronary cineangiograms is a reproducible method for the assessment of the physiological importance of coronary artery obstructions. Short-, medium-, and long-term investigations of the functional results of interventions such as pharmacological therapy or revascularization can be performed reliably with this technique. PMID- 2968197 TI - Cellular mechanisms of myocardial infarct expansion. AB - Infarct expansion is acute regional dilatation and thinning of the infarct zone. There are several possibilities for the mechanism of this alteration in cardiac shape: thinning could be caused by 1) cell rupture, 2) a reduction in the intercellular space, or 3) stretching of myocytes or 4) slippage of groups of myocytes so that less cells are distributed across the wall. To determine the relative contributions of these cellular mechanisms of wall thinning and dilatation, detailed study of transverse histological sections of rat hearts with infarct expansion was performed 1, 2, and 3 days after coronary ligation. The number of cells across the wall was determined in six regions within, adjacent to, and remote from the infarct. Cell counting was performed so that the total number of cells across the wall and the number of cells per unit length (cell density) across the wall were determined. The transmural cell count and the cell density were correlated with the wall thickness in each region. Myocyte cross sectional areas and sarcomere lengths were also measured. The results from the infarct expansion hearts were compared with those of sham-operated control hearts that had been similarly analyzed. To ensure that mechanisms identified in the rat were applicable to human infarct expansion, five hearts from patients who died within 3 days of infarction and two hearts from patients without coronary disease were studied histologically in a similar fashion. Wall thinning occurred in all regions of the rat infarct expansion hearts compared with controls (p less than 0.0001) but, as expected, was most pronounced in the infarct zone. A decrease in the number of cells across the wall accompanied the wall thinning at each site (p less than 0.0001), and this change in cell number was highly correlated with the changes in wall thickness (r = 0.915, p less than 0.001). Cell density increased from controls only within the infarct zone (p less than 0.001) and accounted for at most 20% of the thinning in that region. The change in cell density was attributable to both cell stretch (measured by increased sarcomere length and decreased myocyte cross-sectional area) and a decrease in the intercellular space. A similar strong correlation between wall thinning and decreased number of cells across the wall was identified in the human hearts (r = 0.94, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968198 TI - A case of generalized granuloma annulare responding to hydroxychloroquine. PMID- 2968200 TI - Humoral and cellular immune responses by normal individuals to hepatitis B surface antigen vaccination. AB - The kinetics of the cellular and humoral responses of 30 recipients of hepatitis B vaccine were studied. All individuals exerted an HBsAg blastogenic response sometime throughout the study period but the maximum response was detected on day 28 and 56. The removal of CD8+ cells enhanced significantly the HBsAg response at the times tested, whereas treatment with anti-CD4, anti-CD8, C' and anti-CD4+ C' had no effect. Vaccination also led to the depression of phytohaemagglutinin (PHA) blastogenic response. This response was maximally suppressed 4 to 8 days after immunization at least for the primary and secondary responses and 28 days after the third dose of vaccine. The humoral response to HBsAg was detected only after the second dose of vaccine was given. The results suggest that a CD8+ cell controls the magnitude and intensity of the HBsAg blastogenic response, which may help to explain why several investigators had not been able to detect this response in hyperimmunized individuals. Primary immunization with HBsAg does lead to an expansion of B memory since a secondary response anti-HBsAg was observed. PMID- 2968199 TI - The influence of cyclosporin A on cell-mediated immunity. PMID- 2968202 TI - Peripheral blood dendritic cells in persons with AIDS and AIDS related complex: loss of high intensity class II antigen expression and function. AB - The antigen-specific immune response in HIV sero-positive individuals is depressed or absent. This may be due in part to abnormal co-operation between T lymphocytes and antigen presenting cells (APC). We have isolated from the peripheral blood of healthy heterosexuals and patients with persistent generalized lymphadenopathy (PGL) and AIDS, cells of low density (LDC) with dendritic morphology. These cells are known to be potent APC. The expression of two cell surface antigens on these cells, namely 63D3 (a monocyte related antigen) and Class II antigens was examined. LDC from controls and patients with benign, non-progressive PGL (type A) were found to show biphasic expression of Class II antigens. By contrast, the high intensity Class II expression seen on a small proportion of 63D3 negative cells from controls and patients with PGL type A was absent in patients with PGL type B (showing subtle signs of progressive immunodeficiency) and AIDS. The loss of this population of dendritic cells was reflected in the absence of stimulator activity in autologous and heterologous mixed lymphocyte reactions. Thus, it is possible that the loss of these dendritic cells may contribute to the profound immunological abnormalities seen in AIDS. PMID- 2968201 TI - Defective T-cell colony formation and IL-2 receptor expression at all stages of HIV infection. AB - The T-cell colony assay is a highly sensitive measure of immunological dysfunction. The present study evaluated this in vitro response in asymptomatic HIV-infected homosexuals, those with chronic adenopathy as their only clinical manifestation and patients with either ARC or AIDS. The mean colony count in antibody-positive asymptomatic individuals was significantly reduced when compared to either heterosexual controls or antibody-negative homosexuals. Furthermore, there were no differences in the responses of these antibody positive individuals and those with chronic lymphadenopathy as their only clinical manifestation. By contrast, patients with AIDS or ARC showed a profound defect; this suggests that the colony assay can detect a functional gradient across the spectrum of HIV infections. Colony growth was correlated with the absolute number of T-helper cells and the ability of PHA-stimulated lymphocytes to express IL-2 receptors; no correlation was found with the number of suppressor/cytotoxic cells or in vitro production of IL-2. Recent HIV seroconverters had normal colony counts but impaired ability to express IL-2 receptors. These data suggest a sequential loss of T-cell function as a result of HIV infection; the earliest manifestations are impaired expression of IL-2 receptors and reduced proliferative responses, as measured in the colony assay. PMID- 2968204 TI - Immune complexes and erythrocyte CR1 (complement receptor type 1): effect of CR1 numbers on binding and release reactions. AB - We performed experiments to investigate whether immune complexes opsonized with C3b and iC3b transferred from CR1 on one erythrocyte to CR1 on others, and studied the effect of variation in erythrocyte CR1 number on the transfer reaction. We used populations of cells of different blood groups to study this phenomenon which were separated by differential agglutination with monoclonal anti-group antibodies. The rate of transfer of immune complexes between erythrocytes was related to CR1 concentration of both donor and recipient cells; fastest transfer occurred from donor cells of low CR1 numbers to recipient cells of high CR1. These results were not explained by a difference in the binding constant of immune complexes to erythrocytes bearing different numbers of CR1. In the absence of factor I, complexes partitioned between erythrocytes according to their relative concentrations of CR1 with no release of complexes into solution. In serum, the proportion of complexes bound to donor and recipient erythrocytes was similarly related to their respective CR1 numbers with progressive release of complexes into solution. Erythrocyte CR1 may act as a dynamic buffering system which prevents immune complexes that have bound complement from fixing to vascular endothelium. PMID- 2968203 TI - Defective autologous mixed leukocyte reaction in newly diagnosed type 1 diabetes mellitus. AB - We studied autologous mixed leukocyte reaction (AMLR) in type 1 (insulin dependent) diabetes mellitus (IDDM) patients, their healthy siblings and healthy schoolchildren, Blood samples from the patients were drawn within 1 week after hospitalization and 2 months later. AMLR was significantly depressed in the patients when compared to healthy siblings or other healthy controls. In addition, the mean AMLR responsiveness of the healthy control group exceeded that of healthy siblings. The production of IL-2 in AMLR was impaired in the patient group and the defective AMLR could be restored by addition of exogenous IL-2 in 7 10 patient cultures. However, in 3-10 patients addition of IL-2 induced no enhancement of proliferation. While the patients in general had raised levels of activated T lymphocytes these three patients had higher numbers of activated T cells than other patients. Defective AMLR and presence of activated T cells may be related and may play a role in the pathogenesis of IDDM. PMID- 2968206 TI - Occupational dermatoses. PMID- 2968205 TI - Immunomodulatory properties of cimetidine in ARC patients. AB - The immunomodulatory potency of cimetidine, a histamine H2 receptor antagonist, was investigated in 33 AIDS-related complex (ARC) patients performing detailed immunological and clinical evaluations. Cimetidine was administered orally in daily doses of 1200 mg for a period of 5 months with an interruption of therapy after the first 3 months for an interval of 3 weeks. Significant (P less than 0.05) elevations of immunoglobulins (IgG, IgA), complement C4, B-lymphocytes, and OKT4+ (helper/inducer) cells were found after cimetidine intake. The in vitro lymphocyte proliferative response to plant mitogens was significantly increased, and the in vivo cell-mediated hypersensitivity reaction assessed by intradermal application of seven recall antigens improved significantly. These effects were both reversible with the discontinuation of cimetidine and reproducible with repeated administration of the drug. Clinical data such as performance status, body weight, and fever were influenced favorably (P less than 0.05) by cimetidine. The frequency of diarrhea and the lymph node size were also diminished significantly. The data suggest that cimetidine may at least partially restore immunofunctions in AIDS-related complex. PMID- 2968207 TI - Looking at work dermatologically. AB - I hope that this brief review has been sufficient to prove the importance that looking at patients' work should have in the investigation of suspected occupational dermatoses. For the dermatologist subspecializing in occupational cases, visits are, I believe, a continuing necessity in the weekly routine. For all other dermatologists, even occasional visits can make an important contribution to the accuracy with which diagnoses and advice on prevention can be provided in suspected cases of occupational dermatoses. PMID- 2968208 TI - How to protect the hands. AB - Hand protection entails many problems. There is a wide variety of individual differences in the types of skin among human beings, and an even wider variety of chemicals to be handled and working methods to be learned in various workplaces. There are great differences in the degree of experience and education among job applicants and thus in their ability to understand the importance of instructions on safe working methods and the use of personal protective equipment. Therefore, proper employee selection is an important and demanding task for occupational health care personnel and dermatologists. Despite these complexities, an appropriate means of hand protection is essential in the prevention of many skin disorders and injuries. PMID- 2968209 TI - Prevention of occupational skin disease through use of chemical protective gloves. AB - Selection of chemical protective gloves for use against industrial liquids in the controlled workplace is accomplished by risk analysis, in which the appropriate physical and chemical glove properties needed by the worker to perform the job are determined. Candidate protective gloves are then subjected to chemical permeation testing. Three representative case studies illustrate risk analysis and glove selection. PMID- 2968211 TI - Occupational nail disorders. AB - Evaluation of dystrophic nails must include a thorough assessment of the patient's occupation. Unless this is done, it will not be possible to ascertain the correct diagnosis, and without this, successful therapy is impossible. The spectrum of clinical change exhibited by the nails is limited so that every effort must be made to ensure accuracy. PMID- 2968210 TI - Medicolegal aspects of occupational skin diseases. AB - Skin diseases, including chemical burns, are the most frequently reported of all occupational illnesses. More than 90 per cent of work-related dermatologic problems are contact dermatitis, and there is rarely anything about the location and appearance of these lesions to differentiate clearly from a dermatitis of nonoccupational origin. A complete and time-consuming history must therefore be taken, including documentation of nonwork activities. PMID- 2968212 TI - Patch testing in occupational hand dermatitis. AB - Patch testing is valuable for the management of most forms of hand eczema; it tends to be diagnostically useful, however, only in the nonendogenous forms. Historical and morphologic clues that distinguish hand eczema of endogenous from that of exogenous causes can increase the yield of valuable information from patch testing. Knowledge of the occupations at risk for skin disease and the time required for irritant and allergic contact dermatitis to develop will improve history taking--the key to suspecting an occupational component. PMID- 2968213 TI - Contact urticaria from rubber gloves. AB - Surgical rubber (latex) gloves can cause contact urticaria and even more severe allergic symptoms; the frequency of this allergy may be as high as 3 per cent among hospital employees. Those at high risk are surgeons and instrument nurses, most of whom are atopics. Clinical history of immediate rubber-glove allergy can be misleading, but the diagnosis is easily confirmed by prick, use, and RAST tests. Sensitized persons manage well by using plastic undergloves or vinyl gloves. PMID- 2968215 TI - Dermatitis in the printing industry. AB - Dermatitis is not an uncommon occurrence in the printing trades. The risk of irritation or allergy varies with the different types of printing methods. Once established to the point of disabling the worker, the occupational dermatitis tends to persist. PMID- 2968214 TI - Dermatitis in rubber manufacturing industries. AB - This review describes the history of rubber technology and the manufacturing techniques used in rubber manufacturing industries. The important aspects of the acquisition of allergic and irritant contact dermatitis within the industry are presented for the reader. PMID- 2968216 TI - Predisposing factors in occupational skin diseases. AB - Work-related skin disorders do not appear randomly; they are the result of certain predisposing factors in the environment and the host. These factors can be isolated and recognized, and awareness of these factors by employers and affected employees should have a significant impact on work-related skin disease. The role of a dermatologist in providing this information is essential. PMID- 2968217 TI - Dermatologic problems of office workers. AB - Occupational dermatoses among office personnel are infrequent. Nevertheless, the dermatologist should be aware of the various suspected causes of these dermatoses (poor ventilation, visual display terminals, paper, etc.), since some patients will attribute their cutaneous as well as extracutaneous symptoms to office work. PMID- 2968218 TI - Is the skin affected by work at visual display terminals? AB - According to some reports, mainly from Norway and Sweden, visual display terminal work is suspected of causing skin rashes. In three different studies, we have tried to elucidate the question, and the results indicate that there might be a relation between VDT work and aggravation of some common skin diseases such as rosacea, seborrheic and atopic dermatitis, and acne. Whether this depends on physical, chemical, or psychological factors is still unknown. PMID- 2968219 TI - Prevention of occupational skin disease. AB - Each occupational setting has unique risks for occupational skin disease based on the materials used, the workplace environment, and the skin type of each worker. Virtually all programs for the prevention of skin disease can be designed around the principles outlined in this article. Most problems can be eliminated by well designed prevention programs. All workers are entitled to a safe working environment that reduces to the extent possible the risks of acquiring occupational skin disease. PMID- 2968220 TI - Physicochemical methods for detection of contact allergens. AB - In the same way that it often is necessary to perform chemical and biological analysis of skin and body fluids in the management of skin disease, it may be necessary to analyze the chemical environment. Some analyses, such as those for nickel, chromate, and formaldehyde, require only simple tests. Others require complex and expensive equipment and specialized skills. In the future, it may become necessary to install some of this equipment for routine use in dermatology clinics. In the meantime, the dermatologist needs to be aware of the possibilities. PMID- 2968221 TI - The relationship between hypertrophy and dilatation in the postmortem heart. AB - Confusion may exist at the time of postmortem examination as to whether the diseased heart is dilated, hypertrophied, or both. Ventricular dilatation and ventricular hypertrophy were therefore evaluated by cardiac partition techniques in 441 subjects at autopsy to determine their relationship. Specific weight and surface area of each ventricle were obtained and patients were divided into categories of disease. Wall thickness measurements, a parameter routinely used in the ordinary autopsy, were found to be unreliable in defining hypertrophy. Ventricular surface area (an index of dilatation) was highly correlated with ventricular weight in most disease categories. Exceptions were cardiomyopathy and aortic stenosis, in which hypertrophy predominated. We conclude from these data that dilatation and hypertrophy occur proportionately in the postmortem heart in most disease categories except in cardiomyopathy and aortic stenosis. These findings clarify the relationship of dilatation and hypertrophy at the time of autopsy in most cases. Therefore, uncertainty as to whether cardiac dilatation or hypertrophy is present or which predominates is usually related to the inability to assess these states critically at the time of autopsy when the ordinary pathological methods are used. PMID- 2968222 TI - [A study on Lp(a) lipoprotein in various diseases of the nervous system]. PMID- 2968223 TI - Reversal of tropicamide-induced mydriasis by thymoxamine eye drops. AB - In a study in 12 healthy volunteers, local instillation of thymoxamine eye drops (0.5%), completely reversed the mydriasis produced by tropicamide (0.5%), but only incompletely that by tropicamide (1.0%). The difference between these effects was statistically significant (p less than 0.025). The thymoxamine eye drops were well tolerated. PMID- 2968224 TI - U4 and U6 small nuclear ribonucleoprotein particles. 1. Fractionation and characterization of snRNPs containing U4 and U6 RNAs. AB - The isolation of U4 and U6 small nuclear ribonucleoprotein particles (snRNP) was undertaken, since there has been no reliable method for their fractionation established. The procedure subjects a nuclear extract from HeLa cells to several types of ion exchange chromatography at moderate ionic strength, electrophoresis on agarose gels, transfer of the particles on DEAE cellulose paper, and elution with ammonium chloride. The purified U4 and U6 snRNPs contain U4 and U6 RNAs, respectively, and a set of six polypeptides, present in U4 RNPs and seven polypeptides in U6 snRNP particles. Five major proteins with molecular masses of 70, 64, 47, 40.5 and 24 kD are common to both assemblies. In addition U4 RNPs contain one polypeptide of 25.1 kD which is unique for this class of particles, while U6 RNPs possess two polypeptides with molecular masses of 31 and 18 kD which are not present in U1, U2 or U4 snRNPs. The U4 and U6 RNP particles thus fractionated retain their antigenicity as judged by their reaction with auto antibodies from patients with lupus erythematosus. PMID- 2968225 TI - Quantitative analysis of mouse interferon-beta receptor-mediated endocytosis and nuclear entry. AB - The dynamics of internalization and transport of murine interferon-beta (MuIFN beta) to the cell nucleus was studied in cultured L929 mouse fibroblasts employing postembedding, ultrastructural quantitative immunocytochemistry. Cells preincubated with MuIFN-beta at 4 degrees C were transferred to 37 degrees C for various times, and then fixed and embedded in Lowicryl K4M. Internalized native IFN molecules were identified and quantified on the surface of ultrathin sections with monoclonal (MAb) or polyclonal (PAb) anti-IFN antibodies and a colloidal gold-protein A (CGPA) conjugate. Immunocytochemically recognizable MuIFN-beta molecules entered the cytoplasm by receptor-mediated endocytosis and were translocated via transport vesicles to the nuclear envelope. IFN molecules entered the nucleus through nuclear pores and accumulated maximally in the dense chromatin within 3 min of incubation of the cells at 37 degrees C. The data presented constitute the first direct observation of the dynamics of the extremely rapid transfer of ligand to its intracellular target, in this case, the cell nucleus. PMID- 2968226 TI - Rapid hemodynamic improvement in right ventricular infarction after coronary angioplasty. AB - A patient is described where the hemodynamic disturbance caused by a right ventricular infarction was promptly corrected after coronary angioplasty (PTCA). This indicates that reperfusion may be useful in managing hypotension due to predominant right ventricular infarction. PMID- 2968227 TI - Ipratropium in patients with COPD receiving cholinesterase inhibitors. AB - Three patients with chronic obstructive lung disease (COPD) and myasthenia gravis whose pulmonary symptoms were worsened by therapy with cholinesterase inhibitors were improved by inhaled ipratropium bromide. Two had increases in FEV1 (19 percent, 35 percent) and specific conductance (106 percent, 81 percent) and reductions in dyspnea. The third had no change in airflow with ipratropium, but improved due to decreased bronchial secretions which had limited the use of cholinesterase inhibitors. In contrast, beta agonist bronchodilators had no effect in any of these patients. This experience suggests that ipratropium may be the bronchodilator drug of choice in patients with obstructive lung disease aggravated by cholinesterase inhibitors. PMID- 2968228 TI - [Radiologic diagnosis and treatment of Budd-Chiari syndrome]. PMID- 2968229 TI - [Drug rashes caused by ampicillin]. PMID- 2968230 TI - [Epidemiologic study on dermatitis of workers exposed to N'N-methylene-bis-(2 amino-1,3,4-thiadiazole) and changes in dermal pigment in children in the neighborhood of the factory]. PMID- 2968231 TI - [Experimental study and clinical use of the bovine pericardium in the repair of abdominal defects]. PMID- 2968232 TI - Catecholamine and endorphin responses to delivery in term and preterm lambs. AB - A marked increase in catecholamines and endorphins at birth has been described in animals and man. Because the factors which regulate catecholamine secretion are incompletely understood and because it has been suggested that endogenous opiates are important in the regulation of catecholamine secretion, we designed studies to compare changes in plasma catecholamines at birth with simultaneously measured endorphins. Catecholamines and beta-endorphin-like immunoreactivity (beta-ELI) were measured at birth in term (145 days) and preterm (130 days) lambs. Preterm lambs were given natural sheep surfactant intratracheally to prevent respiration failure. Following umbilical cord cutting, there was a marked increase in circulating norepinephrine (NE) and epinephrine (E) levels. The peak preterm NE (2.2 +/- 0.3 ng/ml at 1 h) was greater than the peak term NE (1.0 +/- 0.2 ng/ml at 5 min). The peak preterm E occurred later and also was greater than the peak term E (4.0 +/- 0.5 ng/ml at 1 h vs. 1.3 +/- 0.4 ng/ml at 15 min, respectively, p less than 0.01). Baseline beta-ELI in term animals (767 +/- 15 pg/ml) was greater than preterm (456 +/- 12 pg/ml). Following cord cutting peak beta-ELI in term animals rose to 867 +/- 201 pg/ml compared to a peak beta-ELI in preterm animals of 1,866 +/- 450 pg/ml. These results and their significance for neonatal adaptation to extrauterine life are discussed. PMID- 2968233 TI - [Morphometric study on sensitization by hematoporphyrin derivative in Ehrlich ascites tumor cells irradiated by gamma-ray]. AB - In this paper, light and electron micrographs of Ehrlich ascites tumor cells radiated by gamma-ray and influenced by hematoporphyrin derivative (HPD) were analysed by morphometric method. It was found that in these cells, the domain of microfilaments treated by HPD expanded extensively, which enhanced the damage of cells induced by gamma-ray. Twenty five minutes after radiation, the cellular fragments increased, resulting in a decrease in the mean diameter of cells and an increase of nuclear volume density and nucleocytoplasmic ratio. In addition, ribosomes on the rough endoplasmic reticulum surfaces fell off increasingly and the quantity of smooth endoplasmic reticula increased. PMID- 2968234 TI - [Allergic contact eczema caused by an acetone-formaldehyde condensate in billboard workers]. AB - We report two cases of occupational contact dermatitis in billboard workers due to employment of a new paste additive. Clinical picture, patch test result, histological pattern of the test reaction and course of the disease indicate that the observed eczema are manifestations of contact allergy and do not represent irritant-toxic dermatitis. The glue is a hydrous solution of a heterogeneous mixture of condensation products of acetone and formaldehyde with a small part of free acetone and formaldehyde. Exact determination of the allergen and identification of the substance group failed. The allergen seems to be a strong sensitizer. PMID- 2968235 TI - An outbreak of acneiform eruption in a polyvinyl chloride manufacturing factory. AB - An outbreak of acneiform eruptions occurred in 14 workers in a polyvinyl chloride (PVC) manufacturing factory. The cause was probably a combination of heat, high humidity and irritation from PVC powder. PVC slurry, scales and powder, to which the workers were exposed, were not comedogenic on rabbit ear. Lowering the temperature and humidity by airconditioning and improving ventilation, together with reduction of dust level at the workplaces, eliminated further outbreak's. PMID- 2968236 TI - Hormonal regulation of type I insulin-like growth factor receptors of Leydig cells in hypophysectomized rats. AB - The effects of hCG and various pituitary hormones on type I insulin-like growth factor (IGF) receptors of purified Leydig cells of hypophysectomized rats were studied. The number of type I IGF receptors of Leydig cells obtained from hypophysectomized rats (18.0 +/- 1.5 fmol/10(6) cells) was lower than that in normal rats (54.6 +/- 5.3 fmol/10(6) cells; P less than 0.05). After a single administration of hCG (10 U, ip), specific binding of [125I]IGF-I to purified Leydig cells increased 3-fold. Scatchard analyses of the binding data suggested that increased binding was the result of an increase in receptor number, whereas binding affinity remained unaltered. Type I IGF receptor increased within 12 h and remained persistently elevated 96 h after hCG treatment. Administration of hCG (10 U, ip) daily for 5 days increased type I IGF receptor levels to 73.2 +/- 8 fmol/10(6) cells (P less than 0.001). FSH caused a small but significant increase in type I IGF receptors. Concomitant administration of FSH and hCG further enhanced IGF-I-binding capacity. IGF-I-binding affinity of Leydig cells treated with FSH or FSH plus LH was not significantly different from that in the control hypophysectomized rats. Daily administration of GH for 5 days also upregulated type I IGF receptors, whereas PRL had no effect. FSH, GH, and PRL administration had no effect on serum testosterone levels. Serum testosterone levels increased to 3.99 +/- 0.35 ng/ml after 5 days of treatment with hCG. Concomitant administration of FSH and hCG caused a further increased in serum testosterone levels (6.13 +/- 0.46 ng/ml; P less than 0.01). The present study suggests that type I IGF receptors of Leydig cells can be up-regulated by LH, FSH, and GH. However, hCG/LH seems to be the most important factor in maintaining and regulating type I IGF receptors of Leydig cells. Steroidogenic and growth promoting effects of hCG and pituitary hormones on Leydig cells may be mediated by increased type I IGF receptors. PMID- 2968237 TI - Thyroxine-induced cardiac hypertrophy: time course of development and inhibition by propranolol. AB - To investigate the mechanism of thyroid hormone-induced cardiac hypertrophy, we have studied the in vivo changes in cardiac size and total myocardial content of both membrane and cytoskeletal enzymes in the rat after the administration of excess thyroid hormone. In response to 50 micrograms T4/day, there is a significant increase in heart rate and heart work associated with an increase in total heart size and protein content. Measurements of the specific activity of Na,K-ATPase and p-nitrophenol phosphatase demonstrate a small but significant increase in specific activity, while the specific activity of myosin ATPase is unchanged. To further probe the mechanism for T4-mediated hypertrophy we studied the in vivo effects of beta-adrenergic blockade on rat heart size. When animals were treated with both T4 and propranolol (10 mg/animal.day) cardiac hypertrophy was prevented. Propranolol alone at this dose did not affect heart rate, heart weight, or serum levels of T4 and T3. The present data suggest that 1) the hypertrophic response of the myocardium to excess thyroid hormone involves cytoplasmic as well as membrane proteins, 2) the increase in total myocardial protein, which can be blocked by propranolol, is indirectly mediated by increases in cardiac work rather than a direct effect of thyroid hormone. PMID- 2968238 TI - The adrenal capsule alters the response of zona glomerulosa cells to atrial natriuretic peptide. AB - Atrial natriuretic peptide (ANP) is a potent inhibitor of potassium-stimulated aldosterone secretion. In the present study, we observed rat alpha ANP to inhibit aldosterone secretion stimulated by 10 mM potassium with an IC50 of 0.15 +/- 0.02 nM (mean +/- SE) in dispersed rat adrenal glomerulosa cells. However, when rat adrenal capsules, which contain the zona glomerulosa, were superfused in vitro, ANP had no effect on aldosterone secretion. Superfusion with 10 mM potassium increased aldosterone secretion 3- to 4-fold above baseline. Addition of 10 nM ANP to the superfusate did not lower potassium-stimulated aldosterone secretion. When this same ANP-containing superfusate was incubated with dispersed adrenal glomerulosa cells, potassium-stimulated aldosterone secretion was inhibited by 90%, proving sustained biological potency of the superfused ANP. Incubation of [125I]iodo-ANP with adrenal capsules for 60 min resulted in 83% degradation of [125I]iodo-ANP, whereas no detectable degradation was observed with dispersed adrenal glomerulosa cells. Removal of blood from the adrenal capsules or culturing the capsules for 48 h did not render them responsive to superfused ANP. In contrast, superfusion of 0.1 mM cycloheximide inhibited potassium-stimulated aldosterone secretion by 90%. These results suggest that the adrenal capsule contains an ANP-degrading enzyme(s). This enzyme may be produced by adrenal glomerulosa cells. The local existence of a degrading enzyme for ANP may allow the zone glomerulosa to regulate its response to ANP. PMID- 2968239 TI - Modulation of atrial natriuretic factor by thyroid hormone: messenger ribonucleic acid and peptide levels in hypothyroid, euthyroid, and hyperthyroid rat atria and ventricles. AB - The effect of thyroid hormone on atrial natriuretic factor (ANF) production was investigated in hypothyroid, euthyroid, and hyperthyroid rats by measuring levels of ANF mRNA and ANF in myocardium. ANF mRNA was quantitated by dot blot hybridization, and ANF by specific RIA. Relative ANF mRNA concentrations (ANF mRNA to 18S RNA) were determined for right atria, left atria, and ventricular apices. The total chamber content of ANF mRNA was estimated (concentration X total chamber RNA) and used as a measure of each tissue's synthetic capacity. For both atrial tissues, ANF mRNA contents were significantly higher in hyperthyroidism. In right atria, mean ANF mRNA contents in hypothyroidism and hyperthyroidism were 41% and 176%, respectively, of that in euthyroidism (P less than 0.05, by analysis of variance). Left atrial ANF mRNA contents in hypothyroidism and hyperthyroidism were 94% and 272%, respectively, of the euthyroid value (P less than 0.05). In contrast, atrial ANF mRNA concentrations did not differ significantly between thyroid states. In ventricle, ANF mRNA content and concentration were both correlated with serum T4 concentration. Ventricular ANF mRNA contents in hypothyroidism and hyperthyroidism were 31% and 178%, respectively, of that in euthyroidism (P less than 0.02). The concentration of ventricular ANF mRNA was also significantly increased in hyperthyroidism (P less than 0.05). Tissue content of ANF increased in the hyperthyroid right atria and decreased in the hyperthyroid left atria and ventricles. These observations suggest that increased ANF production by both atria and, to a lesser extent, by the ventricles contributes to the higher circulating ANF levels reported in hyperthyroidism. Furthermore, hyperthyroidism is associated with a specific increase in ventricular ANF mRNA expression as has been observed in other conditions causing ventricular hypertrophy. PMID- 2968240 TI - Laparoscopic findings in two adults with chickenpox. AB - We report the laparoscopic findings in two previously healthy adults with chickenpox and hepatic involvement. Both patients were admitted to hospital with severe upper right quadrant abdominal pain coinciding with the skin rash. Liver function tests were abnormal. In case 1, round lesions with a necrotic center, 2 mm in diameter, in both hepatic lobes were observed. Morphologically they resembled the rash on the skin. Numerous granular lesions were also demonstrated. In case 2, laparoscopic examination was performed recently when the cutaneous rash had faded. Granular images were scarce; there was enlargement of the liver capsule with small depressed areas of pink color. These lesions were considered similar to those found in case 1, although in a more advanced stage of scarring and resolution. Both patients made a good recovery. PMID- 2968241 TI - Laparoscopic cytology--an evaluation. AB - Cytological sampling by abrasion and/or aspiration was done in a consecutive series of 927 patients at the time of laparoscopy. Cytological examination confirmed the diagnosis of malignancy in 312 out of 335 cases of primary or secondary liver cancer (93.1%). Biopsy was positive in 262 out of 303 patients (86.4%). The combined use of cytology and biopsy provided a positive diagnostic accuracy of 97.6%). The cytological report was positive in 215 out of 239 (89.9%) proven extrahepatic malignancies (gallbladder, gynecological, peritoneal). Biopsy was positive in 180 out of 199 cases (90.4%). The combined use of cytology and biopsy provided an accuracy of 95.8%). There was only one false positive diagnosis of malignancy. The sensitivity of laparoscopic cytology was 93.1% and its specificity 100% in liver disease. Sensitivity was 90.3% and specificity 99.4% in other types of abdominal cancer. In these series, cytological examination added 15.6% positive results (liver) and 11.7% (other lesions) to those of biopsy, which was not performed in 89 patients in whom it was considered too risky or technically too difficult. PMID- 2968242 TI - "Adhesionectomy" under visual control in conjunction with laparoscopy- application of microwave tissue coagulation. PMID- 2968244 TI - Postnatal development of Ca2+-sequestration by the sarcoplasmic reticulum of fast and slow muscles in normal and dystrophic mice. AB - Ca2+-uptake activities of the sarcoplasmic reticulum (SR) were determined with a Ca2+-sensitive electrode in homogenates from fast- and slow-twitch muscles from both normal and dystrophic mice (C57BL/6J strain) of different ages. Immunochemical quantification of tissue Ca2+-ATPase content allowed determination of the specific Ca2+-transport activity of the enzyme. In 3-week-old mice of the dystrophic strain specific Ca2+ transport was already significantly lower than in the normal strain. It progressively decreased with maturation and reached only 40 50% and 30-50% of the normal values in fast- and slow-twitch muscles of adult dystrophic animals, respectively. Tissue contents of calsequestrin were reduced in both types of muscle leading to an increased Ca2+-ATPase to calsequestrin protein ratio. Equal amounts of the Ca2+-ATPase protein (detected by Coomassie blue staining of polyacrylamide gels) were present in SR vesicles isolated by Ca2+-oxalate loading from adult normal and dystrophic fast-twitch muscles. However, the specific ATP-hydrolysing activity of the enzyme was approximately 50% lower in dystrophic than in normal SR. The reduced ATP-hydrolysing activity was correlated with decreased Ca2+-transport activity, phosphoprotein formation and fluorescein isothiocyanate labeling as determined in total microsomal and heavy SR fractions. Although the Ca2+ and ATP affinities of the enzyme were unaltered, its ATPase activity was reduced at all levels of ATP in the dystrophic SR. Taken together, these findings point to a markedly impaired function of the SR and an increase in the population of inactive SR Ca2+-ATPase molecules in murine muscular dystrophy. PMID- 2968243 TI - Mutagenicity of products generated by the reaction between several antiparasitic drugs and nitrite. AB - Drugs containing secondary aliphatic amines, heterocyclic nitrogen, or secondary aliphatic amido groups (chloroquine, dehydroemetine, mebendazole, and piperazine) and pyrimidine derivatives such as pyrantel pamoate were reacted in vitro with sodium nitrite at pH 3.7 and became mutagenic for Salmonella typhimurium strain TA1535. The products derived from the nitrosation of chloroquine and dehydroemetine required metabolic activation by mammalian hepatic S9 to be mutagenic. The N-nitroso derivatives of mebendazole, piperazine, and pyrantel pamoate were mutagenic with and without S9, although more activity was noted in the presence of S9 with the nitrosated compounds formed from mebendazole and piperazine. Under identical conditions, no mutagenic products were detected from quaternary ammonium salts such as bephenium hydroxynaphthoate or drugs containing tertiary heterocyclic amino groups, such as iodochlorhydroxyquin. PMID- 2968245 TI - The isolation and characterisation of human atrial natriuretic factor produced as a fusion protein in Escherichia coli. AB - Human atrial natriuretic factor [ANF(1-28)] has been isolated from a fusion protein produced in Escherichia coli. ANF(1-28) was linked to a naturally occurring E. coli protein, chloramphenicol acetyltransferase, via unique cleavage sequences susceptible to either human thrombin digestion, or the chemical action of 2-(2-nitrophenylsulphenyl)-3-methyl-3'-bromoindolenine (BNPS-skatole). The linker sequences were Gly-Val-Arg-Gly-Pro-Arg and Trp respectively. The liberated ANF was purified by reversed-phase HPLC. Optimised cleavage conditions released 5 10% (by mass) of the maximal yield of ANF(1-28) from the fusion protein with the thrombin-susceptible linker, whilst a 2-5% (by mass) yield was observed from the fusion protein with the tryptophan linker after BNPS-skatole treatment. The purified cleavage products were biologically active and shown to comprise intact ANF(1-28). Fast-atom-bombardment mass spectrometry confirmed [MH]+ of 3079 m/z, consistent with ANF(1-28). PMID- 2968246 TI - Expression of atrial natriuretic factor as a cleavable fusion protein with chloramphenicol acetyltransferase in Escherichia coli. AB - Recombinant fusion proteins containing human atrial natriuretic factor, ANF(1-28) joined to chloramphenicol acetyltransferase (CAT) via cleavable linker sequences have been produced in Escherichia coli. The linker sequences were designed to allow the release of authentic ANF(1-28) following proteolytic cleavage by enterokinase or thrombin, or chemical cleavage with 2-(2-nitrophenylsulphenyl)-3 methyl-3'-bromoindolenine. Proteins, containing ANF(1-28) fused to the carboxyl terminal region of CAT (using the ScaI restriction site in the cat gene), were largely soluble in E. coli and were obtained in higher yield than analogues containing ANF(1-28) linked to shorter CAT sequences. The longer derivatives also retained CAT activity allowing subsequent purification by affinity chromatography. PMID- 2968247 TI - Risk factors for stroke in chronic atrial fibrillation. AB - Atrial fibrillation is associated with an increased risk of stroke. Different risk factors may be of importance regarding stroke incidence and mortality in atrial fibrillation. Retrospectively, we studied 786 patients with chronic atrial fibrillation, 229 with rheumatic heart disease and 557 without. In all, 127 patients had stroke (16%). Using Cox's proportional hazard model for failure-time data with age, gender, etiology, degree of heart failure and cardiac enlargement as explanatory (independent) variables, only etiology (rheumatic heart disease) was a significant risk factor for stroke (P less than 0.006). Significant risk factors for death in 653 patients without stroke were age (P = 0.000) and congestive heart failure at the onset of atrial fibrillation (P = 0.000). The need to identify other risk factors for stroke in patients with atrial fibrillation is emphasized, for selecting patients at high risk for prophylactic treatment with anticoagulants or aspirin. PMID- 2968248 TI - Immediate PTCA after successful thrombolysis with intracoronary streptokinase, three years follow-up. A matched pair analysis of the effect of PTCA in the randomized multicentre trial of intracoronary streptokinase, conducted by the Interuniversity Cardiology Institute of The Netherlands. AB - Immediate PTCA following thrombolysis with streptokinase was performed in 46 out of 533 patients enrolled in a multicentre randomized trial of early reperfusion in patients with acute myocardial infarction. Additional effects of PTCA in patients with a residual diameter stenosis in the infarct-related coronary artery of 70% or more after thrombolysis were compared with successful thrombolysis alone in a matched pair analysis. Thirty six pairs of patients were formed identical with respect to the infarct related coronary artery, presence or absence of previous myocardial infarction, total ST segment elevation on the ECG at admission to the trial, and delay between onset of symptoms and hospital admission. PTCA after thrombolysis did not lead to additional limitation of infarct size, nor to further preservation of left ventricular function. Infarction rate during the three-year follow-up was 14% after PTCA versus 30% after thrombolysis alone (P = 0.05). Similarly, patients had less angina or heart failure after PTCA, since on average 128 out of 156 weeks follow-up were symptom free, while this was only 102 weeks after thrombolysis alone (P = 0.03). Immediate PTCA after thrombolysis with intracoronary streptokinase seems to prevent recurrent ischemia and reinfarction. Further studies should address the proper indication and timing of PTCA after thrombolysis. PMID- 2968249 TI - Sustained improvement in myocardial perfusion four to six years after PTCA in patients with a satisfactory angiographic result, six months after the procedure. AB - Of 46 consecutive patients who underwent percutaneous transluminal coronary angioplasty (PTCA) from April 1980 to August 1982 and who had a satisfactory angiographic result six months later, one died of cancer and 41 had a detailed evaluation with exercise single-photon emission computed tomography (SPECT, 40 patients) or repeat coronary angiography (one patient), 48-78 months after the procedure. During follow-up, two patients had recurrent angina due to progression of coronary artery disease requiring a second PTCA procedure of another coronary artery; one of them later had a limited myocardial infarction in an area supplied by the vessel initially dilated. At follow-up, only one patient had definite angina. All but one patient had a negative exercise test. Myocardial perfusion during exercise in the PTCA-related area, assessed by SPECT, was normal in 90% of the patients and showed a limited defect due to reversible ischaemia in the remaining four (10%). It is concluded that patients with a less than 50% stenosis six months after PTCA show sustained improvement in their functional status and myocardial perfusion, four to six years after the procedure suggesting continued patency of the coronary artery. PMID- 2968250 TI - Advanced angioplasty. Meeting of the European Society of Cardiology Working Group on Myocardial Function. 5-6 June 1986, Lausanne, Switzerland. Proceedings. PMID- 2968251 TI - Restenosis after coronary angioplasty: review of the literature. AB - The average restenosis rate reported so far in the literature is just below 30%. Although restenosis correlates well with the recurrence of symptoms, the two factors are not identical. The incidence of myocardial infarction during the first 2 years after coronary angioplasty is 4% and the incidence of death is 2%. These two cardiac events are rarely the first symptom of restenosis. Restenosis, therefore, is not primarily a life threatening disease but still deserves prompt evaluation and correction. Restenosis is stenosis-related rather than patient related. Thus, restenosis rate per patient increases with the number of lesions or arteries treated. Restenosis rates vary considerably with centres. Serial analyses of restenosis rates at individual centres revealed that the restenosis rates remained constant at a centre-specific level. Differences in case selection and particularities in data definition and analysis may account for both these observations. There is no sound evidence that procedural factors (balloon size, number, duration, or pressure of inflations, etc.) or drug regimens are capable of reducing the recurrence rate. All risk factors for restenosis identified so far are difficult to influence. Extinguishable factors such as smoking seem of little importance in this particular problem. Efforts to find ways of reducing restenoses after coronary angioplasty are commendable and necessary. Their chance of success, however, is small. 'Old customers' will continue to represent 20-30% of the clientele for coronary angioplasty. Their risk for failure and complications is small, but they do carry a considerable risk of restenosis. PMID- 2968252 TI - Restenosis and repeat coronary angioplasty in Geneva. PMID- 2968253 TI - Restenosis after balloon dilatation of coronary stenosis, multivariate analysis of potential risk factors. AB - This study was undertaken to analyze change in stenosis caliber up to six months after PTCA with respect to regression or progression as well as to detect factors which possibly influencing the restenosis rate. A computer assisted system with high accuracy was used for two-dimensional quantitation of stenosis. A linear multivariate analysis was applied to quantitative and qualitative angiographic data as well as to clinical findings obtained before, immediately after and six months post-PTCA in 95 consecutive patients in whom 101 stenoses were dilatated. All patients were on a standard medical regimen of aspirin or coumadin and nifedipine. After six months, 56 patients showed a change in minimal stenosis area (mSA) of less than 1 mm2 (no progression), 33 patients showed a decrease in mSA of greater than 1 mm2 which rendered the stenosis with greater than 70% luminal reduction, and 12 patients showed a decrease in mSA of greater than 1 mm2 which did not, however, result in high-grade luminal narrowing. With regard to factors capable of affecting restenosis rate, there was no relationship between extent of dilatation achieved, local dissection, stenosis configuration or localization, calcification, patient age, sex, duration of symptoms, overweight, cholesterol, triglycerides, HDL, LDL, smoking, hypertension or diabetes. However, a relationship was found between the discontinuation of aspirin or coumadin as a result of GI side effects or bleeding (2% no progression; 20% progression). Thus, antiplatelet therapy appears to be important with respect to long-term results after PTCA. PMID- 2968254 TI - The inducibility of coronary artery spasm after successful PTCA. AB - An ergonovine test (0.2 mg ergonovine i.v.) was performed on 32 consecutive patients showing a good long-term result 3 to 6 months after successful PTCA. In seven patients (22%) a significant spasm could be induced. Six of the seven patients complained of atypical chest pain as compared to two of 25 patients with a negative test. This difference was statistically significant (P less than 0.01). These results implicate that a higher vasomotor tone at the site of previous stenosis is responsible for atypical chest pain in patients after successful PTCA and good angiographic long-term follow-up. The application of long-term nifedipine and nitroglycerin in this patient group seems to be indicated. PMID- 2968255 TI - Laser assisted coronary angioplasty. PMID- 2968256 TI - Prevention of coronary restenosis by stenting. AB - Balloon angioplasty fails to provide acceptable long-term results for a significant proportion of patients. An intravascular mechanical support, developed with the aim of preventing restenosis and acute closure of diseased arteries after transluminal angioplasty, was implanted in 44 patients (39 male and five female), aged from 35 to 70 years (mean 56 years) with documented restenosis of native coronary artery (41 stents) and bypass grafts (12 stents). In the group of bypass graft patients there was no local restenosis and no major complication. In patients in whom stents were placed in native coronary arteries, the complication rate was higher (two patients died after coronary bypass surgery). One patient died suddenly at home. Except for one patient, in whom a new lesion developed proximally with extension into the stent, no case of restenosis could be observed. Despite the still relatively high complication rate, we feel that stenting may present a rational approach to the unresolved problem of restenosis after coronary angioplasty. PMID- 2968257 TI - Recurrent restenosis after transluminal coronary angioplasty--dilatation or surgery? AB - In a total of 333 patients who had undergone a first successful transluminal coronary angioplasty (TCA), restenosis occurred in 17% (follow-up angiography was performed in 94% of patients). The restenosis rate was higher in bypass stenoses (45%) and re-opened vessels (54%). Repeat dilatation of restenoses had a high acute success rate (93%) and only few severe complications (2%). In this group the restenosis rate was 33%. Thirteen patients with recurrent restenoses (11 patients with two, and two patients with three recidivations) underwent a total of 41 dilatation attempts. The degree of stenoses (prior to the first TCA, 89%; prior to second TCA, 82%; prior to third TCA, 74%), the number of eccentric stenoses (8, 7, 5, respectively) and the length of the stenotic obstruction (5.2 mm, 4.7 mm, 4.3 mm, respectively) decreased. Accordingly, exercise tolerance improved (99 W, 133 W, 146 W, respectively). To date, follow-up angiography and functional investigations have been performed for 10 out of 13 patients. Good long-term results were observed in seven patients. Further restenoses occurred in three patients. It is concluded that repeat angioplasty is a reasonable therapeutic approach even for patients with recurrent restenoses. PMID- 2968258 TI - Evaluation of renal first pass blood flow with a functional image technique in hypertensive patients. AB - The renal circulation of patients with essential hypertension and renovascular hypertension was evaluated using 99mTc-DTPA. The first renal peak count (the first Cmax; FCmax), time phase distribution (the first Tmax; FTmax), and blood velocity (the FCmax/FTmax) were calculated by digital imaging. This yields a visual image of the renal circulation. We consider that the increase in the renal first pass blood flow in patients with essential hypertension is best observed pixel by pixel. The FCmax and FCmax/FTmax images before and after treatment by percutaneous transluminal renal angioplasty in patients with renovascular hypertension clearly show its therapeutic effect. The FI technique, therefore, has the advantage that it can be performed at the same time as the conventional routine examinations of renal function. This makes it very useful clinically. PMID- 2968260 TI - Increased number of CD4 cells able to bind to natural killer cell targets in the peripheral blood of AIDS related complex patients. AB - CD4 cells forming conjugates with natural killer target cells (K562 cells) were measured in the peripheral blood of anti-HIV antibody positive AIDS related complex (ARC) patients and in three control groups (asymptomatic individuals at risk, normal healthy people and patients with acute hepatitis B). These CD4 cells, which are unable to kill K562 cells, were significantly increased in ARC patients as compared to the control groups. Our data indicate that classical CD4 cells are partially replaced, in ARC patients, by a population of natural killer target binding granular CD4 lymphocytes, and suggest that the functional abnormalities of helper T cells in these patients may be in part a consequence of the relative predominance of these non IL-2 and BCGF producing cells within the circulating CD4 population. PMID- 2968259 TI - 99mTc-MAG3: dynamic studies in patients with renal disease. AB - Animal studies have suggested that 99mTc-mercapto-acetylglycyl-glycyl-glycine (99mTc-MAG3) might be suitable for the determination of the renal plasma flow (RPF) because of its high renal clearance. In this study 131I-orthoiodohippurate (131I-OIH) and 99mTc-MAG3 (labeling always greater than 95%) were administered simultaneously in 11 patients (creatinine clearance ranging from 14 to 130 ml/min per 1.73 m2) to measure effective RPF(ERPF) using the standard technique (UV/P). Glomerular filtration rate (GFR; clearance of 125I-thalamate, 125I-OT) was also measured. The mean ratio of 99mTc-MAG3 clearance to 131I-IOH clearance was 0.55 +/- 0.02 (SEM), P less than 0.01, n = 16, and was independent of GFR and ERPF. To study this difference in renal handling of the radiopharmaceuticals, renal extractions by the right kidney were determined in another six patients after a single shot of the agents. Renal extraction of 99mTc-MAG3 was 0.60 +/- 0.03 after 5 min, and 0.41 +/- 0.08 after 30 min. Renal extraction of 131I-OIH amounted to 0.86 +/- 0.04 and 0.77 +/- 0.03, respectively. Using renal extractions of 0.41 and 0.77, respectively, it appeared that calculated renal plasma flows measured simultaneously with 99mTc-MAG3 and 131I-OIH were similar. Protein binding 30 min after the priming dose was 66% for 99mTc-MAG3 and 47% for 123I-OIH. We conclude that in spite of a high renal clearance (ratio to 125I-OT clearance 2.69 +/- 0.27), 99mTc-MAG3 seems unsuitable for an accurate determination of the RPF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968261 TI - Clinical and endocrine effects of cyproterone acetate in postmenopausal patients with advanced breast cancer. AB - A phase II study with cyproterone acetate (CPA) was done as the primary treatment in female breast cancer patients. Twenty-three patients, mean age 64 years, range 52-75 years, were entered and treated with CPA 400 mg daily. Twenty patients were evaluable and responses were sparse. There was one partial and one complete remission, 17 patients were stable and one patient progressed within 3 months. Side-effects were frequent: five patients complained of nausea, three had severe weight loss, one suffered from depression and seven showed disturbed liver function tests. Six patients had to stop treatment for side-effects, while two other patients were taken off treatment because they developed an acute necrotizing hepatitis. The hepatitis recovered after drug withdrawal in both patients. The serum levels of CPA, cortisol, androstenedione, DHAS, LH, FSH and prolactin were measured during CPA treatment. The levels of cortisol and androstenedione did not change, while LH, FSH and DHAS were suppressed. The DHAS showed an inverse relation to serum CPA concentrations. The prolactin levels rose uniformly. The therapeutic effect of CPA in postmenopausal patients with advanced breast cancer is disappointing, and inferior to that of other progestins. Side effects are frequent, possibly as a result of the high dosage used in this study. The hormonal changes are different from those of other progestins, which may explain the different efficacies. PMID- 2968262 TI - Adjuvant therapy for operable breast cancer with medroxyprogesterone acetate alone in postmenopausal patients or in combination with CMF in premenopausal patients. AB - The present paper concerns two multicenter studies on adjuvant therapy with medroxyprogesterone acetate (MAP) for operable N+ breast cancer. The patients entered the study between April 1979 and March 1986. One hundred and fifty-one premenopausal patients were randomly assigned to receive either polychemotherapy (CMF) or CMF + MAP. One hundred and thirty-eight postmenopausal patients were randomized to receive either MAP h.d. or no treatment. CMF was administered according the following schedule: cyclophosphamide mg 100/ms p.o. 1-4 days; methotrexate mg 40/ms i.v. and fluorouracil mg 600/ms i.v. 1st and 8th days. The cycle was repeated six times every 28 days. MAP was administered at 1000 mg X 2/daily p.o. for 30 days and afterwards 500 mg X 2/daily for 5 months. In the premenopausal study after a median follow-up of 36 months no difference was observed in the incidence of recurrence, site of recurrence, actuarial 5-year disease-free survival (DFS) or overall survival (OS). In the postmenopausal study a statistically significant lower number of recurrences was observed in MAP treatment patients after a median follow-up of 37 months. The effect of MAP was limited to patients with less than or equal to 3 metastatic axillary lymph nodes. In addition, there are suggestions that only patients with ER+ tumors draw some advantage from the treatment. On the other hand, no difference exists in the OS. The treatments were substantially well tolerated. The MAP + CMF regimen induces lower vomiting compared to the CMF alone. The most frequent MAP side-effects were vaginal spotting (16%) and tremors (12%). We conclude that MAP h.d., like tamoxifen and aminoglutethimide, can improve the DFS of operable N+ breast cancer in postmenopausal patients. PMID- 2968263 TI - A randomized comparison of doxifluridine and fluorouracil in colorectal carcinoma. AB - In a randomized study 52 patients with advanced colorectal cancer and measurable lesions were treated with doxifluridine 4000 mg/m2 or fluorouracil 450 mg/m2 i.v. on 5 consecutive days over 3 weeks. None had prior fluoropyrimidines except two who received adjuvant fluorouracil. Partial responses with a duration ranging from 259 to 406 days were observed in five patients treated with doxifluridine and two patients treated with fluorouracil. Toxic reactions were evaluated in 88 doxifluridine courses and 105 fluorouracil courses. The most frequent adverse effects were neurotoxicity (48% of patients) and mucositis (43%) for doxifluridine, leukopenia (48%) and nausea/emesis (37%) for fluorouracil. Mucositis, diarrhea, nausea, emesis and skin reactions were observed in both treatment groups. Fluorouracil produced neurotoxic effects in 26% of patients. Reversible cardiac dysfunctions were observed in four patients treated with doxifluridine, expressed by ectopic ventricular beats (2) precordial pains (1) and ventricular fibrillation (1). This latter toxicity justified the premature interruption of the study. Doxifluridine is an active agent in colorectal cancer. Compared to fluorouracil it produces, when used i.v., a lower myelosuppression and a greater incidence of neurological and cardiac toxicity. PMID- 2968264 TI - A phase II study of doxifluridine in patients with advanced breast cancer. PMID- 2968265 TI - Toremifene, a new antiestrogenic compound, for treatment of advanced breast cancer. Phase II study. AB - Forty-six postmenopausal women with estrogen receptor positive advanced breast cancer were treated with the novel antiestrogen toremifene in this phase II study. The patients had no prior or concurrent hormonal or cytostatic treatment. Sixty milligrams of toremifene was given as a single daily dose for a minimum treatment period of 6 weeks. Eight patients (17%) achieved complete response, 17 (37%) partial response and 12 (26%) showed no change. The median durations of responses were 93, 66 and 24 weeks, respectively. Three patients still continue the treatment in complete response, four patients in partial response. No significant differences in response rates could be seen when related to different estrogen receptor concentrations. The treatment was well tolerated, only two patients had remarkable side-effects; one of the patients interrupted the treatment mainly because of tremor. Our conclusion is that toremifene is an effective, safe and in clinical practice easily applied choice of treatment in estrogen receptor positive advanced breast cancer. PMID- 2968266 TI - Plasma BGP: an indicator of spontaneous bone loss and of the effect of oestrogen treatment in postmenopausal women. AB - One hundred and ninety-one healthy early postmenopausal women, aged 45-54 years, were randomized to 2 years of treatment with (a) percutaneous 17 beta-oestradiol combined with progesterone (n = 29) or placebo (n = 28); (b) oral oestradiol valerate combined with cyproterone acetate (n = 37) or placebo (n = 39); (c) 24R, 25 (OH)2D3 (n = 29) or placebo (n = 29). We measured the plasma bone Gla-protein (BGP), bone mineral content of the proximal forearms (BMC), bone mineral density in the spine (BMDspine) and total body bone mineral (TBBM) in all the women before, and during, the study. In the groups of women receiving the oestrogen preparations, the plasma BGP decreased highly significantly (P less than 0.001) to a premenopausal level. The initial plasma BGP concentration was significantly related to the loss of BMC (P less than 0.001) in the placebo groups. The changes in plasma BGP were an indicator of the oestrogen response on BMC. We conclude that serial determinations of plasma BGP are useful for determination of the effect of oestrogen therapy in groups of patients, and that plasma BGP measured at the time of the menopause indicates what the rate of bone loss will be. PMID- 2968267 TI - A double-blind and cross-over comparison of once daily doxazosin and placebo with steady-state pharmacokinetics in elderly hypertensive patients. AB - The alpha 1-adrenoceptor antagonist doxazosin has been compared with placebo in 40 elderly hypertensive patients (mean age 71.4 years). At the end of 10 weeks once daily treatment with doxazosin the mean 24-h post-dose changes in standing and supine blood pressure compared with placebo were -6.9/-5.6 mmHg (systolic/diastolic) and -6.2/-5.5 mmHg respectively. The reductions in standing and supine diastolic blood pressures were statistically significant compared with placebo. At the end of treatment steady-state pharmaco-kinetics were evaluated in 18 patients. The plasma elimination half-life during the dose interval in these patients was 16.1 h (range 10.1-27.1 h) and the median time to peak plasma concentration was 3 h (range 1-4 h). One patient was withdrawn because of adverse effects (headache, weakness, and sweating) during doxazosin treatment. Once daily doxazosin reduced diastolic blood pressure and was well tolerated in these elderly hypertensive patients. PMID- 2968269 TI - Functional differentiation and repertoire diversification of T cells derived from single progenitor cells. AB - Functions of T cells derived from single progenitor cells were investigated. B10. Thy-1.1 recipient mice were either whole body-irradiated and marrow reconstituted or thymus-shielded, irradiated and marrow reconstituted, and limited numbers (3 x 10(3) or 6 x 10(3] of a 1:1 mixture of bone marrow cells from C57BL/6 and B6.Lyt 2.1 mice were transferred intrathymically (i.t.). Donor-type (Thy-1.2+) cells of the thymus of a small portion of recipients which expressed the phenotype of either Ly-2.2 or Ly-2.1 but not both were regarded to be a clone of T cells derived from a single progenitor cell, and such clones were assayed for polyclonal helper (Th) and polyclonal cytolytic (CTL) activities as well as alloantigen-specific proliferative (mixed lymphocyte reaction; MLR) and CTL activities. Clones taken 4 weeks after transfer (4-week-old clones) which were generated in the thymus of whole body-irradiated recipients showed polyclonal CTL but not polyclonal Th activity, whereas 4-week-old clones generated in the thymus of thymus-shielded recipients showed both polyclonal CTL and Th activities. Similarly, 4-week-old clones generated in whole body-irradiated recipients responded with CTL to alloantigens when induced in the presence of T cell growth factors but not with MLR, whereas 4-week-old clones generated in thymus-shielded recipients showed both MLR and CTL to alloantigens. Repertoire diversification of 4-week-old clones, however, was incomplete, since clones generated in whole body irradiated recipient did not necessarily respond with CTL to all alloantigens examined, and those generated in thymus-shielded recipients did not necessarily respond with MLR to all the antigens. On the other hand, T cell clones were shown to fully mature by 7 weeks after transfer in terms of cell function as well as repertoire diversification. PMID- 2968268 TI - Comparison of the antihypertensive effects of celiprolol and acebutolol. AB - The antihypertensive effects of the new cardioselective beta-blocker celiprolol and acebutolol have been compared. Thirty patients with arterial hypertension WHO Grade I-II were treated in a double-blind fashion with celiprolol or acebutolol. Before starting the treatment and on Days 15 and 29, before the morning dose, blood samples were taken for measurement of the plasma level of celiprolol. At the same times physical examinations, and clinical and urine chemistry analyses were performed. At the 99% probability level both drugs had significantly lowered the systolic and diastolic blood pressures to normal values at the end of the second and fourth weeks. There was no significant difference between their antihypertensive efficacy. The decrease in diastolic blood pressure at the end of the second week was significantly correlated with the reciprocal of the plasma celiprolol concentration at steady-state at the end of the dosage interval. PMID- 2968270 TI - Stimulation of dopamine D-2 but not D-1 receptors reduces immobility time of rats in the forced swimming test: implication for antidepressant activity. AB - The involvement of dopamine D-1 and D-2 receptor mechanisms was investigated in the forced swimming test with rats. d,1-Sulpiride, a D-2 receptor antagonist, reported to reduce desipramine-induced anti-immobility, did not alter the brain levels of desipramine. In addition, the anti-immobility effect of desipramine was not antagonized by SCH 23390, a D-1 receptor antagonist. Amineptine (20 mg/kg i.p., 60 min before testing), a dopamine uptake blocker, and LY171555 (0.2 mg/kg i.p., 60 min before testing), a dopaminergic D-2 stimulant reduced immobility time in the forced swimming test, but benserazide + 1-DOPA (200 mg/kg p.o., 45 min before testing), which increases dopamine release, or SKF 38393A (20 mg/kg s.c., 60 min before testing), a D-1 agent, did not. The anti-immobility effect but not the stereotypy was increased following chronic (21 days) LY171555 (0.1 and 0.2 mg/kg i.p.) treatment. The effect of acute or repeated (7 days) LY171555 (0.2 mg/kg i.p.) treatment was antagonized by 1-sulpiride (50 mg/kg i.p., 90 min before testing), a D-2 receptor antagonist. Neither SKF 38393A (20 mg/kg s.c., 60 min before testing) nor SCH 23390 (0.05 mg/kg s.c., 30 min before testing) modified the acute anti-immobility effect of LY171555 (0.2 mg/kg i.p.) SCH 23390 (0.025 and 0.05 mg/kg) increased the immobility time at doses which decreased motor activity. The increase in immobility time brought about by SCH 23390 was not antagonized by SKF 38393A (20 mg/kg). The findings indicate that activation of dopamine D-2 receptors could reduce immobility time. PMID- 2968271 TI - Different pharmacologic activities for 13-azapinane thromboxane A2 analogs in platelets and blood vessels. AB - A series of 16 13-azapinane thromboxane A2 analogs were synthesized and their pharmacological dissociation constants (Kd) determined for the thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor in both washed human platelets and canine saphenous veins. Twelve of the analogs were antagonists of both U46619 induced platelet aggregation and saphenous vein contraction. The rank order potencies of these analogs were significantly different in platelets compared to saphenous veins. Four of the derivatives were antagonists in the platelets but possessed agonist activity in the vessels. The potency of the analogs and the type of activity (agonist or antagonist) were found to be sensitive to the substitution pattern of the aromatic ring on the bottom side chain. These results further support the notion that platelet TXA2/PGH2 receptors are distinct from that of vascular receptors. PMID- 2968272 TI - SCH 23390--a selective dopamine D-1 receptor antagonist with putative 5-HT1 receptor agonistic activity. AB - The selective dopamine D-1 receptor antagonist SCH 23390 has been tested in vitro in the rat fundus model and in vivo in the electrically stimulated flexor reflex model. In the fundus model, SCH 23390 showed a potent agonistic activity compared to that of different 5-HT receptor agonists. Pindolol, 1-propranolol and pirenperone showed no or only weak inhibition of the SCH 23390-induced contractions in the fundus strip whereas methysergide was a potent inhibitor. The 5-HT3 receptor antagonist ICS 205-930 did not induce an inhibitory effect. In the electrically stimulated flexor reflex model in pithed rats, SCH 23390 induced a marked increase of the reflex. This increase was slightly inhibited by a mixed dopamine (DA) D-1/D-2 antagonist cis(Z)-flupentixol and by a specific DA D-2 antagonist YM 09151-2. Different reference antagonists: bicuculline (GABAergic), propranolol (beta-adrenergic), scopolamine (muscarinic), yohimbine (alpha 2 adrenergic), prazosin (alpha 1-adrenergic) were all without an antagonist effect on the SCH 23390-induced increase of the flexor reflex. Ketanserin, a selective 5 HT2 receptor antagonist, showed a weak and short-lasting inhibition of the SCH 23390 effect in high doses, whereas ritanserin showed only 35% inhibition of the SCH 23390-induced flexor reflex at a dose of 1.3 mumol/kg i.v. The mixed 5-HT1/5 HT2 antagonists methiothepin and metergoline showed a marked inhibitory effect at 2.6 mumol/kg i.v. and 3.1 mumol/kg i.v., respectively (1.3 mg/kg i.v.). These findings suggest that SCH 23390 might possess 5-HT1 receptor agonist activity. PMID- 2968273 TI - Different roles of D-1 and D-2 dopamine receptors involved in locomotor activity of supersensitive mice. AB - Simultaneous stimulation of both D-1 and D-2 receptors is necessary to reverse reserpine-induced akinesia in mice. The effect of supersensitivity on locomotor function was studied in mice after treatment with reserpine for five days. The response of these animals to a mixed D-1/D-2 agonist, pergolide, or to a presynaptic dopamine (DA) releaser, amphetamine, was increased 3-fold, indicating behavioural supersensitivity. Under these conditions, both selective D-1 and D-2 dopamine receptor agonist (SKF 38393 and LY 171555, respectively), given separately, induced locomotor activity. The D-1 antagonist, SCH 23390, inhibited the effect of both SKF 38393 and LY 171555, whereas the DA synthesis inhibitor, alpha-methyl-p-tyrosine (AMPT), and the D-2 antagonist, sulpiride, only abolished the effect of LY 171555. Moreover, AMPT increased the response to SKF 38393 by 80%. The amphetamine-mediated responses were abolished by SCH 23390 whereas sulpiride did not block them. Thus, stimulation of the D-1 receptor seems crucial in supersensitive animals. In another set of experiments, AMPT was administered to mice pretreated with reserpine for five days in order to fully deplete DA stores. Low doses of LY 171555 reduced the response of these animals to SKF 38393 by 60% whereas higher doses potentiated it. This bimodal effect of LY 171555 was blocked by sulpiride. Since amphetamine was unable to reverse the reserpine induced akinesia in these mice, we can conclude that the inhibitory effect of LY 171555 is not related to presynaptic inhibition of DA release. PMID- 2968274 TI - MPP+ depletes retinal dopamine and induces D-1 receptor supersensitivity. AB - Intraocular administration of 1-methyl-4-phenylpyridinium ion (MPP+) to mice resulted in a dose-dependent depletion of retinal dopamine (DA) and 3,4 dihydroxyphenylacetic acid. Pretreatment with benztropine partially prevented the MPP+-induced depletion. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was less active than MPP+ for depleting DA when administered by the same route and pretreatment with deprenyl partially prevented the depletion. Supersensitivity of retinal D-1 receptors resulted following MPP+-induced DA depletion. PMID- 2968275 TI - Comparison of the pyrimidine nucleoside phosphorylase activity in human tumors and normal tissues. AB - The novel fluoropyrimidine, 5'-deoxy-5-fluorouridine (5'-dFUDR) has strong tumor inhibiting effect without severe cytotoxic effects on the normal cells. This pro drug is metabolized by the enzyme pyrimidine nucleoside phosphorylase (PN'ase) to the active form 5-fluorouracil. Comparative determinations of pyrimidine nucleoside phosphorylase activity in tissue extracts revealed a significantly higher activity in the human gastrointestinal cancer tissues (6.84 +/- 0.70 nmol/mg protein) than in normal tissues from the same organ (2.37 +/- 0.21 nmol/min/mg protein). PMID- 2968276 TI - Ultramicroscopic morphometric, biochemical and histophotometric investigations of myocardial biopsies taken from Fallot patients before and after cardioplegia with cold Kirsch's solution. AB - Biopsies taken from the myocardium of 5 patients with Morbus Fallot and from 1 patient with ventricle septum defect were investigated with a combination of morphometric, biochemical and histophotometric techniques in order to study the cardioprotective effect of cold Kirsch's solution. At the final phase of cardioplegia the cardiomyocytes reveal the following alterations: The volume densities of mitochondria, of their degenerated areas and that of cytoplasmic vacuoles show a significant increase whereas that of myofilaments decreases. Cristae and matrix mitochondriales, however, show only moderate alterations without statistical significance. Biochemically the total ATP-concentration and creatine phosphate (CP)-concentration were more or less diminished, in most of the cases the activity of the myosin ATPase was increased, that of the creatine phosphate kinase (CPK) diminished. Compared with the biochemical estimations of the ATPase activity, its histophotometric estimations yielded corresponding results in 2 of 4 cases. In general our findings confirm the cardioprotective effect of Kirsch's solution. The combination of methods used gives more reliable results than one technique alone. PMID- 2968277 TI - The rib cage and abdominal components of respiratory system compliance in tetraplegic patients. AB - The specific compliance of the chest wall and lungs combined was measured in eight patients with stable tetraplegia. Expiration was impeded with a series of spring-loaded resistances, and end-expiratory pressures plotted against changes in chest wall volume at end-expiration. An optical contour mapping system was used to partition changes in chest wall volume into rib cage and abdominal components. These measurements suggest that the compliance of the whole system is reduced by one third in patients with stable tetraplegia, compared with normal subjects. This may be because of abnormal stiffening of the rib cage. PMID- 2968278 TI - Right atrial dilatation increases inositol-(1,4,5)trisphosphate accumulation. Implications for the control of atrial natriuretic peptide release. AB - Stretching the right atrium of isolated perfused [3H]inositol-labelled rat hearts was shown to stimulate the phosphatidyl-inositol turnover pathway as demonstrated by the accumulation of [3H]inositol-(1,4,5)trisphosphate and its degradation products. Stimulation was detectable after 1 min with larger increases observed after 10 or 20 min. These findings demonstrate that the myocardium can respond to dilatation by an activation of the phosphatidylinositol turnover pathway. Such a mechanism has implications for the release of atrial natriuretic peptide following right atrial distention. PMID- 2968279 TI - Atrial natriuretic peptide (ANP) in rat gastrointestinal tract. AB - Acid extracts of rat stomach and small intestine contained 8.6 +/- 3.7 and 39 +/- 15 ng/g of immunoreactive atrial natriuretic peptide (ANP). When studied by gel filtration and reverse-phase high-performance liquid chromatography, the stomach immunoreactivity consisted of multiple components, whereas the small intestine contained mostly proANP and ANP 1-28-like material. These findings indicate that ANP may have a role in the physiology of the gastrointestinal tract, e.g. in the regulation of water and electrolyte absorption. PMID- 2968280 TI - Shift in rat liver glycolysis control from fed to starved conditions. Flux control coefficients of glucokinase and phosphofructokinase. AB - The distribution of flux control coefficients of a glycolytic system in starved rat liver has been determined. The flux control coefficient profile in starved conditions is compared with normal fed conditions showing that in the former phosphofructokinase enhances more than 2-fold its flux control coefficient while glucokinase decreases slightly. The results also show that the starved system has a more complex structure probably because of the greater influence on the flux of the reverse substrate cycles and the synthesis and degradation fructose 2,6 bisphosphate reactions. PMID- 2968281 TI - Urodilatin (CDD/ANP-95-126) is not biologically inactivated by a peptidase from dog kidney cortex membranes in contrast to atrial natriuretic peptide/cardiodilatin (alpha-hANP/CDD-99-126). AB - Atrial natriuretic peptide (CDD/ANP-99-126) is rapidly inactivated by a membrane preparations from dog kidney cortex. Inactivation occurs by cleavage of the ring structure in the position between Cys-105 and Phe-106. A unique proteolytic product separated by HPLC on reverse-phase column appears as a single peak which elutes prior the intact peptide. In contrast, CDD/ANP-95-126 (urodilatin) which is released from the kidney is not destroyed by proteolysis using an identical membrane preparation. PMID- 2968282 TI - The ATP-driven primary Na+ pump in subcellular vesicles of Vibrio alginolyticus. AB - Subcellular vesicles of Vibrio alginolyticus hydrolyze ATP and accumulate Na+ in an ATP-dependent fashion. The Na+ uptake is (i) strongly stimulated by delta psi discharging agents, i.e., the protonophorous uncoupler CCCP or valinomycin + K+ and (ii) arrested by DCCD at a concentration strongly inhibiting ATP hydrolysis. Lower concentrations of DCCD stimulate the Na+ accumulation supported by ATP hydrolysis as well as by NADH oxidation. It is concluded that there is an electrogenic DCCD-sensitive Na+-ATPase in the cytoplasmic membrane of V. alginolyticus. PMID- 2968283 TI - [Nicotinamide in polymorphous light eruption]. PMID- 2968284 TI - [Rosacea-like skin changes following immediate removal of a powder blast injury]. PMID- 2968285 TI - Palmoplantar keratoderma with amyotrophy. AB - Four members of a South African Black family who have palmoplantar keratoderma with amyotrophy are reported. No neuromuscular defects have been found and the muscle thinning appears to result from disuse atrophy. Volar hyperhidrosis, nail abnormalities and in 2 cases knuckle pads, were additional features. The condition appears to be inherited as an autosomal dominant trait. We are not aware of a similar syndrome having been previously reported. PMID- 2968286 TI - Menarche, menstruation, sexual relations and contraception of adolescent females with Down syndrome. AB - Fifteen female adolescents with Down syndrome (DS) and 33 female controls without this syndrome were surveyed on age of menarche, menstruation, sexual relations and contraception. All probands and controls were born between 1 January 1965 and 31 December 1970. The day of census was December 31, 1985. All probands and controls were living permanently in the Danish county of Aarhus on this day. Persons close to the DS patients (parents/foster-parents/permanent staff) were interviewed on the above-mentioned topics. All controls were interviewed themselves. The average age of menarche was 13.6 years for probands and 13.5 years for controls. The average duration of the bleeding was 5.5 days for probands and 5.4 days for controls. The average length of the cycle was 28.3 days for probands and 28.6 days for controls. Thirteen DS patients had not had sexual intercourse (no information was available for 2 probands). Twenty-four controls had and 9 had not had sexual intercourse. There is a significant difference between probands and controls concerning use of oral and other contraceptives. PMID- 2968288 TI - Inhibition by corticosteroids of epidermal growth factor-induced recovery of cyclooxygenase after aspirin inactivation. AB - Cultures of vascular smooth muscle cells superfused with [14C]arachidonic acid synthesized the antiplatelet substance prostacyclin as the major cyclooxygenase product. Prostacyclin synthesis was inactivated by aspirin, which irreversibly acetylates cyclooxygenase. Aspirin-treated cells recovered within 2 h by a process that was blocked by cycloheximide but not by actinomycin D, and that required a serum component identified as epidermal growth factor (EGF). EGF induced recovery of cyclooxygenase was greatly potentiated by type beta transforming growth factor (TGF-beta). Incubation with EGF and TGF-beta in the 0.1-1.0 nanomolar range stimulated cyclooxygenase recovery up to 20-fold without increasing [35S]methionine incorporation into other cell proteins. Induction of cyclooxygenase by EGF and TGF-beta also was prevented by cycloheximide but not by actinomycin D. EGF-dependent recovery was blocked by preincubation with dexamethasone (2 microM), an effect that was duplicated by pure lipocortin (2-4 micrograms/ml). Incubation of membrane preparations from these cells with EGF selectively activated phosphorylation of a 35-kDa cellular protein that comigrated with lipocortin. The results suggest that cyclooxygenase recovery in aspirin-inactivated vascular smooth muscle cells is mediated by an EGF-dependent translational control that is inhibited by corticosteroids. The findings also provide a new mechanism whereby corticosteroids suppress inflammatory prostaglandins. PMID- 2968289 TI - [Transluminal coronary angioplasty. Anatomo-angiographic aspects conditioning the indications]. PMID- 2968287 TI - The correlation between sensory nerve conduction velocities and three metabolic indices in rats treated with streptozotocin. AB - The fastest conduction velocities of the myelinated (A) and unmyelinated (C) sensory nerve fibres were measured in the saphenous nerves of rats made diabetic up to 5 weeks previously by injection of streptozotocin. The conduction velocity of the fastest A-alpha fibres in treated rats fell by 25% compared to control rats. The effect on the slow A-delta fibres was small but C-fibres were not affected. Levels of glycosylated haemoglobin, random plasma glucose, and the net changes in body weights were also measured. There were significant changes in these three metabolic indices among diabetic rats (p less than 0.001) and the three indices were inter-related. There was a good correlation between A-fibre conduction velocity and levels of glycosylated haemoglobin but no significant relationship between C-fibre conduction velocity and this metabolic index. A fibre conduction velocity was statistically more correlated with levels of glycosylated haemoglobin than the other two metabolic indices (p less than 0.01). PMID- 2968290 TI - [The kissing balloon in angioplasty of coronary stenosis involving the bifurcation of a major branch]. AB - A case of severe effort angina treated with Percutaneous Transluminal Coronary Angioplasty (PTCA) is described. The critical coronary lesion involved the left anterior descending coronary artery and the origin of a first diagonal branch. The "kissing balloon" technique was used. After a year, the immediate positive angiographic results remained unchanged. PMID- 2968291 TI - [Acute rickettsial myocarditis. Description of a clinical case and review of the literature]. AB - A case of acute rickettsial myocarditis in a young patient is described. The Authors analyze the clinical features, discuss epidemiologic features and the necessity of treatment with tetracycline. PMID- 2968293 TI - The role of laparoscopy in the diagnosis and management of gastrointestinal disease. Guidelines for clinical application. PMID- 2968292 TI - [Usefulness of specific bronchial provocation tests in the diagnosis of compensable occupational allergic pneumopathies]. AB - The A.A studied the bronchial reactivity of 309 subjects accusing a working respiratory disease. It had been valued the utility of specific provocation bronchial test for diagnosis of allergic bronchopneumopathies. The test, performed with working antigens pool, not only confirms the low percentage of allergic respiratory disease among considered workers (joiners, painters and millers) it also shows its sensibility only in subjects with a sure story of bronchial asthma. The A.A. say, that it's not really necessary to perform the test in all subjects that submit to diagnostic screening for industrial insurance reasons. PMID- 2968294 TI - Contractile function and Ca2+ transport system of myocardium in ageing. AB - Experiments with animals with various species-specific life span (rats, rabbits, cats, dogs) and different models (in situ heart, isolated perfused heart, isolated papillary muscle) have proved the reduction of functional capacity of the ageing heart. Diversely directional age-dependent shifts have been established involving myocardial Ca2+ transport system, i.e. an increase in the rate of Na+-Ca2+ exchange and passive Ca2+ transport across sarcolemma and a decrease in its Ca2+-binding capacity and a decrease in Ca2+ accumulation by sarcoplasmic reticulum and mitochondria (Ca2+ uptake). The experiments revealed a decrease in the Ca2+ ATPase myosin activity in the myocardium of aged animals and absence of age changes in the K+ ATPase activity. The findings obtained suggest that the development in the cardiac contractile function disorders in ageing largely depends on the age-related changes in the Ca2+ transport system. PMID- 2968295 TI - The role of oxygen radicals in immune complex injury. AB - In this review we will summarize our current understanding of the mediation of immune complex induced tissue injury. Comparisons will be made between the mediation of IgG versus IgA immune complex injury with emphasis on the role that reactive oxygen products derived from leukocytic phagocytic cells play in the initiation of the tissue injury. PMID- 2968296 TI - [Structural bases and complications of transluminal balloon angioplasty of the coronary arteries]. PMID- 2968297 TI - [Effect of ischemia and cardioplegia on the hypertrophied myocardium]. PMID- 2968298 TI - [Lumbar intervertebral facet block]. PMID- 2968299 TI - [Electrocardiographic changes and the athlete's heart]. PMID- 2968300 TI - The rearrangement and transcription of alpha and beta T-cell receptor genes in phosphorylcholine-specific suppressor T hybridomas. PMID- 2968301 TI - [The potential thrombolysis under selective infusion of the autologous plasmin (AP) solution]. AB - The lytic therapy using Urokinase (UK) as well as Streptokinase (SK) has a significant risk of complications such as systemic bleeding. We aimed to develop the autologous plasmin (AP) solution as a potential lytic agent and to evaluate its lytic efficacy. Method; The AP solution was aseptically prepared by adding UK to autologous plasma separated by centrifugation (at 4 degrees C, 3,000 rpm, 10 min). The induced plasmin activity of the AP solution was measured by plasminogen free fibrin plate method and spectrophotometric method with substrate S-2251. In vitro study, we made a fibrin clot by adding CaCl2 (1/40 mole, 0.2 ml) to autologous plasma (0.2 ml). The clot weight was measured before and after incubation for 60 min at 37 degrees C to estimate the lytic effects of the AP solution and the UK solution. In animal study, femoral artery of anesthetized mongrel dogs (n = 20) was narrowed by ligation (1 mm in diameter) and the fibrin clot was embolized into this portion. AP (n = 8), UK (n = 6) or saline (n = 6) was selectively injected for 3 min into the arterial lumen, after the temporary flow obstruction was completed by inflation of balloon tip catheter located proximal to the embolized site of the artery. Lytic effects on the embolized fibrin clot were sequentially observed by the extra-vascular ultrasound flow meter (equipped with pencil probe) for 60 min. For this study, the AP solution was prepared by adding dose of 12,000 IU/ml of UK. The same dose of the UK solution was also used as a control.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968302 TI - Actin-like protein from Mimosa pudica L. PMID- 2968303 TI - The transplantation barrier of nude mice. AB - Syngeneic memory cells can be stimulated to yield a secondary immune response after their transfer into irradiated euthymic recipients as well as into young thymusless nude mice. It is shown that nude mice older than twelve weeks of age are not permissive towards memory cell activation as it is found in non irradiated euthymic animals. This barrier to isogeneic or congeneic cells seems to be caused by a pool of cyclophosphamide-sensitive cells. Since young nude mice could be rendered as unpermissive as older nude mice by pretreatment with either PNA-agglutinable thymus cells or nylon-wool passed spleen cells, it is suggested that an increased number of precursor T cells in older nude mice might induce this effect. Further experiments with monoclonal antibodies against the Lyt-1, Lyt-2, and L3T4 marker on T cells indicate that T-helper/inducer activity might be required to establish the "isogeneic barrier" in nude mice. PMID- 2968304 TI - A novel T-lymphocyte molecule that may function in the induction of self tolerance and MHC-restriction within the human thymic microenvironment. AB - T-cell differentiation is known to take place in the thymus, but the precise mechanisms involved remain unresolved. In order to analyse the role of the thymic microenvironment in thymocyte maturation and generation of the T-cell repertoire, we have raised monoclonal antibodies (mAb) to thymic stromal cells, and with these can recognize, in the non-lymphoid component of the thymus, several antigenically distinct compartments. One mAb, MR6, binds to both the cortical epithelium and medullary macrophages/dendritic (M phi/DC) cells in sections of the human thymus. Recently, the molecule detected by MR6 has also been detected at low levels on the surface of T lymphocytes. We now report that this molecule has a relative molecular mass of 145,000 (p145-MR6) and that this appears to be the same for both thymic lymphocytes and stromal cells. Functional studies show that mAb MR6 inhibits both the antigen-specific and the IL-2-induced proliferative response of MHC class II-restricted cloned helper T cells and peripheral blood mononuclear cells (PBMC). These results suggest that the molecule to which mAb MR6 binds could be responsible for the inhibition of T-cell proliferation to self-antigens, and hence may be involved in tolerance induction and MHC restriction. PMID- 2968306 TI - Phenotypic diversity of the CD45 antigen and its relationship to function. AB - Monoclonal antibodies (mAbs) have identified further heterogeneity within the major CD4 and CD8 T-lymphocyte subsets. Antibodies to different polypeptides of the CD45 complex separate two largely non-overlapping subsets that may represent stages in the maturation of T cells or separate functionally distinct subsets. PMID- 2968307 TI - Diltiazem and left ventricular hypertrophy in renovascular hypertensive rats. AB - The effects of diltiazem treatment (40-50 mg/kg/day orally for 8 weeks) of left ventricular hypertrophy on systemic and coronary hemodynamics and mechanical cardiac performance were investigated in renovascular hypertensive rats (Goldblatt, two-kidney, one clip). Systemic and coronary hemodynamics were determined by using radioactive microspheres in conscious, unrestrained rats. Mechanical performance was measured on isolated papillary muscle from the same animal. Nine treated hypertensive rats were compared with control groups: 12 untreated hypertensive and nine sham-operated rats. Diltiazem treatment led to an effective but incomplete control of blood pressure (from 208 +/- 5 mm Hg in the untreated hypertensive group to 155 +/- 3 mm Hg in the treated hypertensive group; p less than 0.01) associated with a significant but incomplete decrease of the left ventricular mass (from 3.10 +/- 0.19 mg/g in untreated hypertensive rats to 2.35 +/- 0.04 mg/g in treated hypertensive rats; p less than 0.01). A close correlation was found between left ventricular mass and systolic blood pressure in untreated, treated, and pooled groups (r = 0.84, p less than 0.001, n = 30). The left ventricular weight to systolic blood pressure ratio was equivalent in all three groups, so that the reduction of left ventricular mass in diltiazem treated rats was commensurate with the reduction of blood pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968310 TI - PPOs for Medicare? PMID- 2968308 TI - Atrial natriuretic factor release by angiotensin II in the conscious rat. AB - Since it was previously reported that atrial natriuretic factor (ANF) may exert an inhibitory effect on renin release, the existence of an Angiotensin II (Ang II)-ANF feedback mechanism was investigated. Male rats were infused intraperitoneally for 7 days with either saline, a nonpressor dose of Ang II (200 ng/kg/min), or a pressor dose (800 ng/kg/min) of Ang II. Systolic blood pressure, plasma ANF, 24-hour urinary sodium excretion, urine volume, and water intake were measured. A significant increase in plasma ANF was observed in the group with a pressor response (blood pressure rose from 89.0 +/- 3.9 to 136.7 +/- 11.4 mm Hg; ANF rose from 36.8 +/- 4.9 to 92.7 +/- 17.7 pg/ml). There was no significant time effect on 24-hour sodium excretion, urine volume, and water intake in both Ang II infused groups. In a second set of experiments, male rats were infused intravenously for 60 minutes with either saline, a nonpressor dose of Ang II (16 ng/kg/min), or a pressor dose (800 ng/kg/min) of Ang II. Left ventricular end diastolic pressure, right atrial pressure, and mean arterial pressure were monitored. There was a significant increase in plasma ANF and left ventricular end-diastolic pressure only with the pressor dose (blood pressure rose from 85.0 +/- 6.1 to 140.0 +/- 5.5 mm Hg; ANF rose from 22.6 +/- 6.0 to 108.3 +/- 47.7 pg/ml; left ventricular end-diastolic pressure rose from 5.3 +/- 5.7 to 20.8 +/- 7.9 mm Hg). No significant modification of right atrial pressure was recorded.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968305 TI - Suppressor T cells, antigen-presenting cells and the role of I-J restriction in oral tolerance to ovalbumin. AB - Suppressor T cells (Ts) and antigen-presenting cell (APC) activity are both important for the induction of systemic tolerance after feeding protein antigens to mice. In this report, we have examined further the nature of the inter relationship between Ts and APC in oral tolerance to ovalbumin (OVA). We found previously that oral tolerance to OVA could prevented by treating mice with oestradiol, and we now report that oestradiol enhances the ability of spleen APC to present OVA to T cells. In parallel, mice treated with oestradiol do not generate the Ts activity normally found after feeding OVA. Treatment of mice with anti-I-J antiserum prevents the induction of both tolerance and Ts activity after feeding OVA, but the suppressor effector cells generated by feeding OVA can not be depleted in vitro by treatment with anti-I-J antibody plus complement. In vivo administration of monoclonal anti-I-A antibody had no effect on oral tolerance to OVA. Our results show that induction of oral tolerance to OVA is an I-J restricted phenomenon and we propose that this reflects an interaction between specific Ts cells and a population of I-J+ cells which we suggest are APC. PMID- 2968309 TI - Dynamic cardiovascular responses to infusions of atrial natriuretic factor in humans. AB - We sought to demonstrate a hypotensive effect from infusions of atrial natriuretic factor (ANF) into humans and to describe the mechanism(s) of this effect. Cardiovascular and hormonal responses to human ANF-(99-126) (125 ng/kg bolus followed by a 30-minute infusion at 25 ng/kg/min) were determined in eight conscious volunteers and compared with responses of eight time-control subjects who received isotonic saline. Baseline levels of ANF (52.8 +/- 5.5 pg/ml) increased 8.8-fold after 30 minutes of ANF infusion but were unchanged in the time controls. Plasma levels of renin, aldosterone, vasopressin, sodium, potassium, and osmolality did not change during infusions. A transient 5% reduction in mean arterial pressure related to a 12% reduction in peripheral resistance was observed 10 minutes after the priming bolus of ANF. This response was not sustained during the remainder of the ANF infusion period, nor did it occur in two additional subjects who received ANF infusions without the priming bolus. Steady state responses consisted of significant reductions in central venous pressure (15%), stroke volume (13%), and cardiac output (10%), but no reduction in blood pressure. Plasma norepinephrine levels and peripheral resistance increased (34% and 9%, respectively) during ANF administration. These data indicate that steady state responses to ANF in humans consist of decreases in cardiac filling pressures, which reduce cardiac output, unload cardiopulmonary baroreceptors, and activate the sympathetic nervous system. Blood pressure is well maintained despite striking increases in plasma ANF. PMID- 2968312 TI - The use of regression analysis to determine hospital payment: the case of Medicare's indirect teaching adjustment. AB - Although the intention of Medicare policy makers was to use only "legitimate" cost differences to adjust teaching hospital payment rates under the prospective payment system (PPS), the actual calculations implicitly recognized others. In this paper I question whether these implicit costs should be passed through, and whether regression analysis was used correctly to establish reimbursement rates for hospitals under PPS. Conducting regression analyses on a more inclusive set of variables than the Health Care Financing Administration (HCFA) used to establish its adjustments, I conclude that costs in teaching hospitals rise only 3.15% for each .1 increment in the resident-to-bed ratio, far lower than HCFA's original 11.59% adjustment and substantially lower than the current adjustment of 7.7% for indirect costs. PMID- 2968311 TI - Market responses to HMOs: price competition or rivalry? AB - Although competition for consumers is increasing in the health care sector, there is disagreement about whether it is resulting in cost containment, as its supporters have argued it would. In part this stems from a confusion between price competition, which under ideal circumstances leads to the production of services at the lowest possible cost, and nonprice competition--or rivalry--which under many circumstances will lead to increased costs. In this paper, I examine the evidence about the competitive response to the growing presence of health maintenance organizations in the health care marketplace. The available evidence suggests that providers are responding not with classical cost-containing price competition but, instead, with cost-increasing rivalry, characterized by increased expenditures to promote actual or perceived product differentiation. PMID- 2968313 TI - The nursing shortage: do we dislike it enough to cure it? AB - The current, and rapidly worsening, nursing shortage is likely to continue in the absence of fundamental changes in nursing education and salaries. Because both hospitals and nurses themselves seem to benefit in the short term from the current education and salary structure, I suggest that the fundamental change necessary to ensure an adequate supply of high-quality nurses in the future will have to come from outside these two constituencies. In this paper, I propose a long-range solution that comprises three basic proposals: 1) Create a nursing education system that clearly differentiates, in terms of salary and certification, between programs that educate registered nurses and associate degree nurses and programs that educate bachelor's and advanced-degree nurses. 2) Expand the nursing salary structure to reward long-term, career-track nurses. 3) Assign nursing responsibilities according to nurses' educational and certification qualifications. PMID- 2968314 TI - What influences a state's approach to Medicaid reform? AB - Four states--Arizona, California, New York, and Pennsylvania--undertook major Medicare reforms in the early 1980s based on competition, price negotiation, regulation, and diagnosis related groups. To increase our understanding of what led to these reforms, we analyzed data from interviews with representatives of state executive and legislative branches and from providers and others involved in the reforms, and examined published and unpublished information on the reforms. We identified seven factors influencing choice of Medicaid reform: the crisis that triggered reform, the purposes of reform, the locus of the innovation (legislative versus executive), the power and views of key provider groups, state officials' perception of providers and recipients, the scope of the change implemented, and reform as a reflection of past state practices. We discuss these factors extensively. PMID- 2968315 TI - Factors that contribute to Medicare HMO risk contract success. AB - In 1985, federal legislation was implemented that allowed HMOs to contract with the Health Care Financing Administration for Medicare enrollees on a risk basis. This change stimulated the development of risk contractors and a growth in HMO Medicare enrollees. This paper examines selected factors that contributed to large Medicare risk contract enrollment in four geographical locations: Los Angeles, Minneapolis-St. Paul, New York City, and Portland, Oregon. The primary types of factors examined were: HMO market conditions, HMO characteristics, and HMO marketing strategies and practices. The market competition varied substantially across the four areas. HMO plan age, total number of HMO enrollees, low premium, and the use of multimedia marketing strategies were positively associated with large Medicare risk contract enrollment. PMID- 2968316 TI - Do HMOs reduce hospitalization of terminal cancer patients? AB - The ability of health maintenance organizations to contain the costs of their members' health care, largely through reduced hospitalization, has been documented. There are, however, no studies to investigate whether HMOs realize cost savings specifically for their terminally ill members. In this study, we examined hospital utilization and costs in a matched-pair comparison of HMO members and nonmembers under the age of 65 in Monroe County, New York, who died of cancer during 1976-1982. We found that the HMO members used somewhat fewer hospital days and had lower hospital costs than nonmembers, but the differences were not statistically significant. We also found significantly greater variance in the costs of care for nonmembers than for HMO members. PMID- 2968317 TI - Utilization of inpatient services under shortened lengths of stay: a neonatal care example. AB - In the last several years many hospitals have experienced a significant reduction in average length of stay (LOS). We know relatively little about whether such reductions are likely to be accompanied by proportional reductions in the utilization of all inpatient services or whether services are merely condensed into a shorter time frame. Average patient severity may well rise as a result of shortened LOS, causing daily resource consumption to rise. In this paper, however, we hypothesize that patients with shorter LOS consume significantly fewer resources. Our empirical results support the hypothesis for some, but not all, of the services were examined. PMID- 2968318 TI - Common sense on extending DRG concepts to pay for ambulatory care. AB - The development of Medicare's prospective payment system has led to considerable interest in expanding case-based payment to other forms of care, including ambulatory care. In applying approaches developed in one form of care to another, one must consider whether the same characteristics and conditions apply. This paper reviews the similarities and differences between ambulatory and inpatient care in light of available research. It identifies critical issues that should be considered in designing ambulatory payment reform and draws conclusions on the appropriateness of extending case-based payment principles to ambulatory care. PMID- 2968319 TI - An assessment of hospital acquisition prices. AB - Although much has been written about hospital acquisitions by investor-owned systems, little attention has been paid to how the systems determine the prices they are willing to pay for acquired hospitals. In this study, we attempted to ascertain whether certain market and management variables explain the purchase price of 37 hospitals acquired by investor-owned systems during the period 1978 1984. Our findings suggest that the systems were willing to pay a higher price per bed for more highly utilized facilities in high-income community settings, but paid less for older, aging facilities that were more likely to require major capital improvements after acquisition. PMID- 2968320 TI - Role of L3T4+ T cells in host defense against Histoplasma capsulatum. AB - Cell-mediated immunity is critical in host resistance against the pathogenic fungus Histoplasma capsulatum. To explore the role of L3T4+ T cells in protection of mice against H. capsulatum infection, we examined the effect of in vivo treatment with anti-L3T4 monoclonal antibody (MAb) GK1.5 on the course of murine disseminated histoplasmosis. Treatment with anti-L3T4 antibody caused a profound and selective depletion of L3T4+ T cells that was associated with a significant increase in the number of H. capsulatum CFU recovered from the spleens of mice infected for 1 week. In addition, none of the infected mice treated with MAb GK1.5 survived a sublethal challenge with H. capsulatum yeasts. Histopathological examination of spleens from mice infected for 1 week revealed the presence of granulomatous inflammation in mice depleted of L3T4+ T cells and in infected controls. However, silver stains demonstrated that spleens of infected mice given MAb GK1.5 contained a greater number of yeasts than did spleens from infected controls. MAb GK1.5 did not cause reactivation of infection when administered for 2 weeks beginning 4 weeks after inoculation of Histoplasma yeasts. MAb GK1.5 did not alter the functional properties of murine macrophages as measured by antigen presentation, production of interleukin-1 in response to lipopolysaccharide, and phagocytosis of H. capsulatum yeasts. These results suggest that the L3T4+ T-cell subset is an essential constituent of the cell-mediated immune defense against H. capsulatum infection. PMID- 2968322 TI - Commuting--a further stress factor for working people: evidence from the European Community. II. An empirical study. AB - This report summarizes the main results of research promoted by the European Foundation for the Improvement of Living and Working Conditions, concerning the impact of commuting on the health and safety of workers. An empirical study, carried out among 1167 industrial Italian workers, shows that "commuters" (workers whose journey from home to work usually does not take less than 45 min in each direction) experienced a more stressed life-style than did "non commuters" (whose journey does not take more than 20 min). Commuting appears for many workers to be a necessity which is imposed by external factors, such as the housing market and job opportunities. Commuting is shown to interfere with patterns of everyday life by restricting free-time and reducing sleeping time. A majority of commuters use public transport mainly because of cost. Public transport commuters have problems due to more changes between modes, idle waiting times and delays leading to late arrival at work. Inside transport modes, commuters suffered discomfort as a result of overcrowding, microclimatic conditions, noise and vibrations. Commuters also reported higher psychological stress scores, more health complaints, essentially of psychosomatic nature, and greater absenteeism from work due to sickness. Commuting, in addition to shiftwork, further increases sleep problems, psychosomatic complaints and difficulties with family and social life. Women commuters were at a greater disadvantage than men, having more family difficulties, more travelling complaints and higher absenteeism. PMID- 2968323 TI - Effect of acute physical exercise on lymphocyte subpopulations in trained and untrained subjects. AB - To clarify the difference in immunity between untrained subjects and well-trained athletes, the number of total leukocytes (WBC), lymphocytes, and neutrophils, percentages of various lymphocyte subpopulations (OKT3, OKT4, OKT8, Leu7, OKla1), and the levels of lymphocyte transformation response to phytohemagglutinin (PHA) were determined in five untrained male subjects and six male athletes before, immediately after, and 24 and 72 h after acute physical exercise at 60% of VO2max for 2 h. Exercise produced a significant rise in the number of WBC, lymphocytes, and neutrophils in both groups. Immediately after exercise, the percentage of OKT3 or OKT4 positive cells had significantly decreased in both groups, whereas that of OKT8 positive cells had markedly increased only in the athletes. Neither group showed any change in the percentage of OKla1 positive cells. In both groups, the response of lymphocytes to PHA immediately after exercise was significantly lower than before, 24 h and 72 h after exercise. The level of Leu7 positive cells rose remarkably immediately after exercise in the athletes, but not significantly in the untrained subjects. These results suggest that an increase in Leu7 positive cells provides added host defense capacity in trained athletes during periods of stress which impair T-lymphocyte function. PMID- 2968321 TI - Deficiency of interleukin-2 activity upon addition of soluble egg antigen to cultures of spleen cells from mice infected with Schistosoma japonicum. AB - Schistosoma japonicum-infected C57BL/6 mice show similar dynamics of hepatic granulomatous inflammation (HGI) and delayed hypersensitivity (DH) elicited by soluble egg antigens (SEA) which reach peak levels at 9 weeks of infection and then spontaneously regress. The in vitro SEA-induced proliferation of spleen cells (SC) from infected animals attained its high point and then declined when SC from 5-week-infected mice were used. The present study determined the dynamics of interleukin-2 (IL-2) production by SEA-challenged SC from infected mice in an attempt to link the level of IL-2 production to the spontaneous regression of the aforementioned T-cell-mediated immune responses. The production of IL-2 by SEA stimulated SC reached its peak when cells from 7-week-infected mice were challenged at least 2 weeks after the peak of the proliferative response, but declined at about the same time as the HGI and DH responses. Therefore, the decline in IL-2 activity cannot alone explain the diminished proliferative response but could account for the reduction in HGI and DH in vivo. Some possible mechanisms that might explain the IL-2 deficiency were examined. This deficiency is not due to the in vitro binding of IL-2 by the SC of infected mice and is, therefore, likely to be due to underproduction of IL-2. Nor is the deficiency explained by reduced numbers of antigen-presenting cells (macrophages and B cells) or of L3T4+ T lymphocytes or by suppression of IL-2 production by macrophages or macrophage products such as prostaglandins. However, suppression of IL-2 production was observed consistently upon coculture of SC from acutely infected mice with SC from mice infected for 10 weeks. The cells which suppress appear to be Lyt2+ T cells. The data are consistent with the hypothesis that suppressor T cells inhibit the production of IL-2 and perhaps of other cytokines or lymphokines and that this suppression explains the spontaneous down-regulation of HGI which occurs during schistosomiasis japonica. PMID- 2968324 TI - Mortality risks in the elderly: five-year follow-up of a total population. AB - The health and social status was assessed by interview for all people aged 75 years and over, living in and around Melton Mowbray. This initial survey took place in 1981 and five years later, data are now available on the mortality status of the original survey population. Significantly lower survival rates were found for various subgroups including those respondents who perceived their health poor, those with high disability scores and severe cognitive impairment and those taking hypoglycaemic agents and diuretics. PMID- 2968325 TI - Medical consequences of a factory closure: illness and disability in a four-year follow-up study. AB - The effect of a factory shut-down on sick leave and disability pensions was investigated in a four-year controlled follow-up study in a general practice setting. The study population consisted of 85 people, 72 women and 13 men, who lost their jobs when a sardine factory near Bergen in Norway was shut down in 1975. The employees of a nearby sister factory' within the same company were chosen as a control population, consisting of 87 people, 66 women and 21 men. The average amount of sick leave for the study group within the first year of follow up showed a twofold increase compared to the controls. The rate of disability pensions, estimated by the life table method with appropriate adjustments, was more than three times higher in the study group than in the control group from the second through the fourth year of follow-up. The results of this investigation provide further evidence in support of a causal relationship between job loss and illness. PMID- 2968326 TI - The influence of medroxyprogesterone acetate (MPA) on hormone parameters in prostatic carcinoma patients. AB - In ten patients suffering from previously untreated, advanced, non-metastatic prostatic carcinoma, the effect of MPA on the hormonal parameters of testosterone and luteinizing hormone (LH) was examined. The dosage of MPA amounted to 500 mg i.m. daily for a week. Thereafter 1000 mg i.m. every four weeks were applied. Full castration could be achieved in all patients, and this effect could be kept up during the time of observation. The tolerance was good, there were no side effects. PMID- 2968327 TI - Echocardiographic tests in chronically haemodialysed patients. AB - Fifty-two patients kept alive by chronic dialysis have been subjected to echocardiography carried out by means of a Picker Echoview equipment with M-mode 2D technique. The revealed alterations were suggestive of hypertropic, dilatative uraemic cardiomyopathy, associated with impaired diastolic compliance. Left auricular dilatation as a source of inference for the degree of compliance diminution was found in a high percentage of patients. PMID- 2968328 TI - Elastic characteristics of the self-expanding metallic stents. AB - We measured elastic properties of the self-expanding metallic (Gianturco) stent using Hooke's law to characterize the stent with respect to its caliber, length, and radius by a coefficient of stiffness. Although this coefficient was independent of the radius of the stent, we found that it was directly dependent on caliber. For a stent of a particular caliber and length, the fractional change in radius determined the force exerted by the stent. For small radial displacements of the stent, the force required to compress it was directly proportional to the radial displacements; for large displacements, the force depended on a power series of the fractional radial displacement. A hyperbolic function was empirically introduced to approximate this type of relationship between force and radial displacement. We calculated tension and pressure exerted by the stent and suggest the use of our findings with normal vessels. PMID- 2968329 TI - Percutaneous transluminal angioplasty in the treatment of lower limb ischaemia. PMID- 2968331 TI - Immunohistochemical localization of beta-endorphin in hyperplastic interstitial tissue of the human testis. AB - The immunohistochemical localization of beta-endorphin in the normal testis (two patients) and in the pathologic testis (two cases of Sertoli Cell Only Syndrome, two cases of Klinefelter Syndrome, two cases of post-orchitis tubular sclero hialinosis) was investigated. No beta-endorphin immunostaining was detected in the normal testis, while positive beta-endorphin immunostaining has been observed in pathologic tissues. These results indicate that, as in animals, beta-endorphin is present in human Leydig cells and may play a local role in regulating male reproductive function. PMID- 2968330 TI - Estimation of tumor oxygenation and metabolic rate using 31P MRS: correlation of longitudinal relaxation with tumor growth rate and DNA synthesis. AB - 31P MRS longitudinal relaxation times (T1) were determined for C3H murine fibrosarcomas (FSaII), and mammary carcinomas (MCaIV). Tumors were implanted in the foot dorsum, and were 100-300 mm3 in volume. T1s were repeated after the animal was allowed to breathe 100% oxygen for 30 min and then again 36-48 hr following 30 Gy. The spectrum were obtained using an 8.5 T spectrometer with a 8 cm bore and a 1.4 cm single turn antenna coil. The 31P relaxation times for untreated tumors in air breathing animals were: 3.78 sec for phosphomonoesters, 4.37 sec for inorganic phosphate (Pi), 2.73 sec for phosphocreatine, 1.37 sec for gamma ATP, 1.14 sec for alpha ATP, and 1.18 sec for beta ATP. The Pi T1s were 4.37 and 4.70 sec in control and irradiated tumors in air breathing animals. Respiration of oxygen for 30 min reduced the T1s to 3.02 and 2.62 sec in control and irradiated tumors respectively. The Pi T1 of an anoxic tumor, determined on an in situ tumor 60 min after death was 5.93 sec. The oxygen breathing induced decrease in the T1 of Pi is unlikely to have been caused by the paramagnetic properties of oxygen alone, and suggests a component of increased magnetization transfer secondary to the ATPase reaction. Oxygen breathing following 30 Gy, resulted in a decreased growth time (800 mm3 endpoint) and an increased proportion of cells in S-phase. These results support the hypothesis that the decrease in Pi T1 measured with oxygen breathing is a measure of tumor oxygen tension and metabolic rate, and suggests that T1 measurement may indirectly predict tumor growth rate and DNA synthesis. PMID- 2968332 TI - LY121019 inhibits Neurospora crassa growth and (1-3)-beta-D-glucan synthase. PMID- 2968333 TI - Current concepts in the treatment of staphylococcal meningitis. AB - As natural host defences are suboptimal in cerebrospinal fluid (CSF), and since antibiotics often penetrate poorly into this body site, meningitis remains one of the most difficult infections to cure. When the responsible pathogen is killed only by a relatively high concentration of an antibiotic, or is susceptible only to drugs with moderate penetration into the CSF, then treatment becomes even more complicated. Therapy of staphylococcal meningitis poses all these problems, and is even further complicated by its frequent development in the presence of a foreign body, such as a ventriculoperitoneal or ventriculoatrial shunt. In this review, we discuss the various options open to the clinician in the treatment of meningitis caused by methicillin-susceptible and methicillin-resistant staphylococci, both in the presence and in the absence of a foreign device. PMID- 2968334 TI - Ovarian pregnancy. PMID- 2968335 TI - Recognitions by the College of Medicine, UAMS: faculty and Caduceus Club. PMID- 2968336 TI - Case report on Escherichia hermannii isolated in an Arkansan. PMID- 2968337 TI - Adenocarcinoma of the breast with metastases to sternum and lungs. PMID- 2968338 TI - Determination of zeranol/zearalenone and their metabolites in edible animal tissue by liquid chromatography with electrochemical detection and confirmation by gas chromatography/mass spectrometry. AB - A sensitive method is described for the determination and confirmation of zeranol and zearalenone, as well as their isomers and metabolites, in edible animal tissue. The analytes are extracted from tissue with methanol, hydrolyzed enzymatically, cleaned up by acid-base partitioning, determined by liquid chromatography (LC) with electrochemical (EC) detection, and confirmed by gas chromatography/mass spectrometry (GC/MS). LC analysis is performed by isocratic elution with a buffered mobile phase using a Nova-Pak reverse-phase C18 column with amperometric EC detection at +0.90 V. Capillary GC/MS analysis of the trimethylsilyl derivatives provides mass spectral confirmations. PMID- 2968339 TI - Replication forks of Escherichia coli are not the preferred sites for lysogenic integration of bacteriophage Mu. AB - The question of whether bacteriophage Mu prefers replication forks for lysogenic integration into Escherichia coli chromosomes was tested by using two different systems. In the first, inactivation of genes was scored in synchronized cultures infected by Mu at various times. No increase in the mutation frequency of a gene was found after infection at the time of its replication. In the second, the composition of colonies formed by bacteria lysogenized by Mu was determined; the newly formed lysogens should give rise to mixed colonies (containing lysogenized as well as nonlysogenized bacteria), uniform colonies, or both, depending on the mode of integration. Both types of colonies were found, and the fraction of uniform colonies was proportional to the relative length of the unreplicated segment of an average chromosome in the culture. The results in both systems clearly preclude the possibility that a lysogenizing Mu integrates with high preference at the chromosome replication forks. PMID- 2968341 TI - Luminescence of deoxyhemocyanin and deoxytyrosinase. AB - The deoxy form of hemocyanins and tyrosinases from certain species displays a weak low-energy luminescence when solutions of the protein are irradiated with light at approximately 290 nm. The emission most likely results from a copper-to imidazole charge transfer state as shown by studies with a synthetic copper(I) complex having three imidazole ligands. PMID- 2968340 TI - Putrescine and spermidine control degradation and synthesis of ornithine decarboxylase in Neurospora crassa. AB - Neurospora crassa mycelia, when starved for polyamines, have 50-70-fold more ornithine decarboxylase activity and enzyme protein than unstarved mycelia. Using isotopic labeling and immunoprecipitation, we determined the half-life and the synthetic rate of the enzyme in mycelia differing in the rates of synthesis of putrescine, the product of ornithine decarboxylase, and spermidine, the main end product of the polyamine pathway. When the pathway was blocked between putrescine and spermidine, ornithine decarboxylase synthesis rose 4-5-fold, regardless of the accumulation of putrescine. This indicates that spermidine is a specific signal for the repression of enzyme synthesis. When both putrescine and spermidine synthesis were reduced, the half-life of the enzyme rapidly increased 10-fold. The presence of either putrescine or spermidine restored the normal enzyme half-life of 55 min. Tests for an ornithine decarboxylase inhibitory protein ("antizyme") were negative. The regulatory mechanisms activated by putrescine and spermidine account for most or all of the regulatory amplitude of this enzyme in N. crassa. PMID- 2968342 TI - Isolation of terminal cisternae of frog skeletal muscle. Calcium storage and release properties. AB - Sarcoplasmic reticulum (SR) terminal cisternae (TC) of frog (Rana esculenta) fast twitch skeletal muscle have been purified by isopycnic sucrose density gradient centrifugation. Biochemical characteristics and Ca2+ release properties have been investigated and compared to those of the homologous fraction of rabbit skeletal muscle TC. The frog SR fraction obtained at the 38/45% sucrose interface appears to be derived from the terminal cisternae region as judged by: (a) thin section electron microscopy showing vesicles containing electron opaque material and squarelike (feet) projections at the outer surface; (b) protein composition (Ca2+ ATPase, calsequestrin, and high Mr proteins); (c) Ca2+ fluxes properties. The content of calsequestrin was higher in frog TC by 50% and the Ca2+ binding capacity (624 or 45 nmol of Ca2+/mg of TC protein, depending upon experimental conditions) was 3-4 times that of rabbit TC. Species-specific antigenic differences were found between junctional SR proteins of frog and rabbit TC. After active Ca2+ preloading in the presence of pyrophosphate (Palade, P. (1987) J. Biol. Chem. 262, 6135-6141), caffeine and doxorubicin elicited Ca2+ release from either TC fraction but with much faster rates in frog TC than in rabbit TC (14 versus 3 mumol of Ca2+/min/mg of protein). The present results provide new evidence for the existence of marked differences in Ca2+ release properties between TC of amphibian and mammalian fast-twitch muscle. Higher Ca2+ binding capacity and faster release rates in frog TC might compensate for the comparably greater diffusion distance being covered by the released Ca2+ from the Z-line to the actomyosin cross-bridges in the A-I overlap region. PMID- 2968343 TI - Stoichiometry and mapping of the nucleotide sites in sarcoplasmic reticulum ATPase with the use of UTP. AB - Purified sarcoplasmic reticulum ATPase was phosphorylated by either ATP or UTP under otherwise identical conditions. Calcium, pH, and nucleotide concentrations were adjusted to permit maximal steady-state accumulation of phosphoenzyme (EP). Either 4 or 8.5 nmol of EP/mg of protein were obtained with ATP or UTP, respectively. Tryptic digestion of phosphorylated ATPase followed by acid gel electrophoresis showed that EP from UTP was on fragment A1, similar to the report in the literature for EP from ATP. Phosphorylation with Pi in the absence of calcium gave EP levels similar to those obtained from UTP. Thus, comparison of EP levels from different substrates measured in parallel in the same preparation reveal that with ATP half of the sites are phosphorylated. Illumination of the ATPase with UV light in the presence of [3H]UTP caused photolabeling of the ATPase at a maximal level of 1 nmol of [3H]UTP incorporated/mg of ATPase. The UTP concentration dependence for photolabeling was the same as that for promoting catalysis. ATP when present in the illumination protected with a competitive pattern against photolabeling with UTP. Tryptic digestion and autoradiography of photolabeled ATPase revealed that UTP was covalently attached to tryptic fragment A2. The data indicate that a peptide sequence of fragment A2 is involved in the binding of the nucleoside moiety of UTP and possibly belongs to the nucleotide domain of the ATPase in addition to the sequence of fragment A1 which contains the phosphorylation residue. PMID- 2968344 TI - Aortic velocity contours at abdominal branches in anesthetized dogs. AB - Pulsed Doppler velocimetry was used to record a grid of velocity waveforms 0.5 cm downstream of the cephalic mesenteric and left renal branches of the aorta in anesthetized dogs. Aortic velocity contours at different phases of the pulse cycle were developed from the grid of waveforms. Transient flow recirculation occurred in the post-systolic phase of the pulse cycle at the aortic wall opposite the branch artery. There was no recirculation at the systolic peak but there was asymmetry of the velocity profile. In contrast, in the in vivo post stenotic velocity field recirculation persists throughout most of the cycle. These results compare well with similar results from pulsatile model studies. Spectral width of the Doppler signal was not increased in any of the velocity waveforms immediately downstream of the branches, this suggests that flow is relatively undisturbed in these locations. PMID- 2968345 TI - Polarized secretion of lysosomal enzymes: co-distribution of cation-independent mannose-6-phosphate receptors and lysosomal enzymes along the osteoclast exocytic pathway. AB - The osteoclast is a polarized cell which secretes large amounts of newly synthesized lysosomal enzymes into an apical extracellular lacuna where bone resorption takes place. Using immunocytochemical techniques, we have localized the cation-independent mannose-6-phosphate (Man6P) receptor and lysosomal enzymes in this cell type in order to determine the expression and distribution of this receptor and its ligands. The results demonstrate that the osteoclast expresses large amounts of immunoreactive cation-independent Man6P receptors, despite the fact that most of the lysosomal enzymes it synthesizes are secreted. The lysosomal enzymes and the receptors are co-distributed along the exocytic pathway, i.e., the endoplasmic reticulum, including the perinuclear envelope, the Golgi stacks as well as numerous small transport vesicles that appear to fuse with the ruffled border membrane. Within the Golgi complex, the receptors and lysosomal enzymes were found distributed in two predominant patterns; (a) in all the cisternae, from cis to trans, or (b) predominantly in cis- and trans-Golgi cisternae, with the middle Golgi cisternae being unstained or depleted in antigen. This pattern suggests that enzymes and receptors traverse the Golgi from cis to trans and preferentially accumulate in cis- and in trans-cisternae. This study therefore suggests that, in the osteoclast, Man6P receptors are involved in the vectorial transport and targeting of newly synthesized lysosomal enzymes, presumably via a constitutive pathway, to the apical membrane where they are secreted into the bone-resorbing compartment. This mechanism could insure polarized secretion of lysosomal enzymes into the bone-resorbing lacuna. PMID- 2968346 TI - Norepinephrine down-regulates the activity of protein S on endothelial cells. AB - The adrenergic agonist norepinephrine is shown to stimulate endothelium to induce protein S release and degradation, leading to diminished anti-coagulant activity and to down-regulation of protein S cell surface-binding sites. Norepinephrine induced release of intracellular protein S was blocked by the alpha 1-adrenergic antagonist prazosin (10(-7) M) but not by the alpha-adrenergic antagonist propranolol (10(-6) M) or the alpha 2-adrenergic antagonist yohimbine (10(-5) M) indicating that this response resulted from the specific interaction of norepinephrine with a class of alpha 1-adrenergic receptors not previously observed on endothelium. Attenuation of norepinephrine-induced release of protein S by pertussis toxin in association with the ADP-ribosylation of a 41,000-D membrane protein indicates that this intracellular transduction pathway involves a regulatory G protein. The observation that protein S was released from endothelium in response to maneuvers which elevate intracellular calcium or activate protein kinase C suggests that the response may be mediated via intermediates generated through the hydrolysis of phosphoinositides. Morphologic studies were consistent with a mechanism in which norepinephrine causes exocytosis of vesicles containing protein S. In addition to release of protein S, norepinephrine also induced loss of endothelial cell protein S-binding sites, thereby blocking effective activated protein C-protein S-mediated factor Va inactivation on the cell surface. Norepinephrine-mediated endothelial cell stimulation thus results in loss of intracellular protein S and suppression of cell surface-binding sites, modulating the anti-coagulant protein C pathway on the vessel wall. These studies define a new relationship between an anti coagulant mechanism and the autonomic nervous system, and indicate a potential role for an heretofore unrecognized class of alpha 1-adrenergic receptors in the regulation of endothelial cell physiology. PMID- 2968348 TI - Microalbuminuria and its associated risk factors in a representative sample of Italian type II diabetics. AB - A multicenter population-based study was undertaken from 1983 to 1985 in 12 diabetes centers in Lombardy, in order to assess the prevalence of microalbuminuria and clinical proteinuria. Out of a total population of 17,704 diabetics, 1,155 were randomly selected within four strata of duration of disease and albumin excretion rate (AER) assessment was carried out in 541 unselected subjects. Albuminuria was evaluated from an overnight urine collection using a radioimmunoassay. The overall prevalence rates were 25.8% for microalbuminuria (30 less than or equal to AER less than 350 micrograms/min) and 3.0% for macroalbuminuria (AER less than or equal to 350 micrograms/min). In Type II diabetes the rate of microalbuminuria increases with duration of disease up to 10 years and then tends to level off. This could be due to the existence of two subgroups of diabetics with different life expectancy and different degree of risk of nephropathy. The Italian prevalence of microalbuminuria appears to be rather similar to that of other studies, while the prevalence of macroalbuminuria is quite low with respect to middle and northern Europe, indicating a rather slow rate of progression to clinical proteinuria. This could be explained on the basis of differing protein intakes. The cases with AER greater than or equal to 30 micrograms/min, greater than or equal to 70 micrograms/min, and greater than or equal to 350 micrograms/min have been compared with matched diabetic controls with AER less than or equal to 15 micrograms/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968349 TI - Prevention of urinary albumin excretion in 6 month streptozocin-diabetic rats with the aldose reductase inhibitor tolrestat. AB - Recent clinical data strongly suggest that elevated urinary albumin excretion (UAE) identifies diabetic subjects at risk of developing nephropathy. Elevated UAE is attributed to increased transglomerular pressure, which is associated with poor metabolic control in rats. Because excess glucose in diabetes is metabolized via the polyol pathway, we were interested in whether the diabetes-induced elevation in UAE in rats could be prevented by inhibiting aldose reductase (AR), the first enzyme in the polyol pathway, with the AR inhibitor tolrestat. In fact, in rats made diabetic with streptozocin (35 mg/kg IV), treatment for 6 months with tolrestat (25 mg/kg/day in the diet) prevented both sorbitol accumulation in the kidney and the increase in UAE. Sorbitol accumulation and the increased UAE were not associated with statistically significant mesangial expansion, and the thickening of glomerular basement membranes was not affected by tolrestat treatment. The authors conclude that the 4.7-fold elevation in UAE in chronically diabetic rats is linked to the increased flux of glucose through the polyol pathway since it was prevented by inhibiting aldose reductase with tolrestat. PMID- 2968347 TI - Preferential expression of fibronectin receptors on immature thymocytes. AB - Fibronectin-adherent (FNR+) thymocytes are enriched for immature (CD4-8-) and large (CD4+8+) cells, and depleted of mature (CD4-8+ and CD4+8-) and nonmature small (CD4+8+) cells. Among purified CD4-8- thymocytes, cells with the surface marker J11d and the IL-2 receptor, which can give rise to all other thymocyte subsets, showed selective attachment to fibronectin. Analysis of FNR+ thymocytes showed that such cells are greatly enriched for cells in cycle. Additionally, FNR+ cells expressed low levels of T cell receptor. These results suggest a role for the fibronectin receptor during the early, proliferative phase of thymocyte differentiation. The data suggest that loss of the fibronectin receptor is a hallmark of cells that have become committed either to functional maturation or to programmed cell death. PMID- 2968350 TI - The selective elimination of anionic immunoglobulins as a parameter of kidney damage in diabetes and diabetic pregnancy. AB - IgG1 and IgG4 have similar molecular weights but differ in pH (about 9 and 4.6, respectively). Their different rates of excretion in the urine of diabetic patients may indicate an impairment of charge selectivity in the kidney filter. Working on this hypothesis, a sensitive new ELISA for the detection of urinary IgG4 has been developed. This method can detect less than 1 ng/ml of this immunoglobulin; total IgG was detected by a RIA method developed by our laboratory. Twenty-eight Type I diabetic patients with or without clinical nephropathy were included in a cross-sectional study. An additional seven diabetic patients were followed over time, and eight diabetic pregnant women were studied during the different trimesters of pregnancy. Whereas both IgG4 and total IgG values were increased in clinically nephropathic patients, levels of IgG4, but not IgG1-3, were enhanced in patients without clinical nephropathy. In the latter group as well, IgG4-positive patients were microalbuminuric; all but one of the remaining patients were IgG4 and albumin negative. There was no significant variation in IgG4 values with time on repeated samples. The increased glomerular filtration rate in diabetic pregnancy did not significantly modify the levels of IgG4 in the urine. These results are in accordance with a selective excretion of this medium to large sized anionic protein (IgG4) in incipient (or stage III) diabetic nephropathy. Urinary IgG4 could be an additional useful marker when studying diabetic patients with early and pre-clinical stages of diabetic nephropathy. PMID- 2968351 TI - Plasma renin and aldosterone in youngsters with insulin-dependent diabetes mellitus. AB - We studied plasma renin activity (PRA) and aldosterone in three groups of subjects. Group 1 consisted of seven Type I diabetics with microalbuminuria (greater than 25 micrograms/min), age 12.0-19.5 yr (mean +/- SD: 15.4 +/- 2.2), duration of disease 6.5-10.1 yr (7.9 +/- 1.9), HbA1c 9.6-16.0% (12.6 +/- 2.9). Group 2 consisted of seven sex and age-matched diabetics, duration of disease 3.7 9.0 yr (6.0 +/- 2.3), HbA1c less than 8%, microalbuminuria less than 10 micrograms/min, and microangiopathy-free. Group 3 consisted of seven healthy subjects. After overnight recumbency the PRA in group 1 patients was significantly higher than that for group 2 (3.926 +/- 4.54 ng/ml/h vs. 1.416 +/- 0.44; p less than 0.05) or for group 3 (3.926 +/- 4.54 vs. 1.11 +/- 0.82; p less than 0.007). After physical exercise the group 1 PRA value (10.199 +/- 9.62) was higher than in either group 2 (2.821 +/- 1.77; p less than 0.005) or group 3 (1.61 +/- 0.803; p less than 0.0006). Poor metabolic control and the presence of microalbuminuria can play a role in perturbations of the Renin-Angiotensin system. The presence of microalbuminuria can be an important indicator of mildly impaired renal function and may influence PRA production. PMID- 2968352 TI - Collisional spectroscopy as a screening procedure for the determination of FFI in complex natural matrices. AB - The build-up of brown products on long-lived proteins could be an important factor in determining long-term diabetic complications. A brown product 2-(2 furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI), present in the acid hydrolysate mixtures of beta-polylysine and beta-albumin has been identified by collisional spectroscopy, using a commercial double-focusing, reverse-geometry mass spectrometer. Using this approach the analyses were carried out without any sample derivatization or need for chromatographic separation procedures. PMID- 2968353 TI - Type I insulin-dependent diabetic patients show an impaired renal hemodynamic response to protein intake. AB - Human kidney responds to a meat meal with an increase in glomerular filtration rate (GFR), but the mechanisms regulating kidney hemodynamics following protein intake are poorly understood in Type I insulin-dependent diabetes. In the present study we investigated GFR response to protein intake (600 gr/1.73 m2 meat meal) in nine normal subjects and 21 Type I insulin-dependent diabetic patients with normal albumin excretion rates as well as proximal tubular sodium reabsorption rates and distal sodium delivery (PRNa and DDNa). The same study was reperformed in normal subjects and diabetic patients, with less than a 5 year diabetes duration, following one week of indomethacin treatment. Normal subjects showed a 38% increase in GFR following protein intake, whereas diabetic patients showed a significantly lower response (18%, p less than 0.01). The response of GFR to protein challenge was negatively related to diabetes duration but not to baseline glomerular filtration rate. Indomethacin treatment completely prevented the protein induced GFR increase in normal subjects but not in diabetic patients. Sodium reabsorption rate was increased following protein challenge both at the proximal and distal tubular level, as was net natriuresis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968354 TI - Effects of nephrectomy and high-protein diets on glomerular hemodynamics and urinary protein excretion in diabetic rats. AB - Renal blood flow, GFR, albumin clearance, and urinary excretion of proteins were assessed in intact control and streptozotocin-diabetic rats (group 1), unilaterally nephrectomized control and diabetic rats (group 2), and nephrectomized control and diabetic rats (divided into high (a) and low (b) glycemia subgroups) fed a 50% protein diet (group 3). After 8 months of diabetes, blood flow did not differ from control rats within each experimental group, although it was increased significantly in both controls and diabetics of groups 2 and 3 versus group 1. GFR in control rats was increased approximately 2x by nephrectomy and approximately 3x by nephrectomy plus the high protein diet. Diabetes increased GFR approximately 50% above control values in group 1; GFR values in diabetic rats of groups 2 and 3a were virtually identical and similar to those of group 2 controls; GFR in group 3b diabetics was increased approximately 1.4x versus group 2 diabetics. 125I-BSA clearance was increased 3.4x in groups 2 and 3 control rats versus group 1 controls. Both nephrectomy and consumption of the high protein diet caused marked increases in 125I-BSA clearance and urinary excretion of albumin in diabetic rats. Urinary excretion of IgG was increased by diabetes in group 1 and remained essentially at this level in groups 2 and 3 diabetic rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968355 TI - Effects of long-term diabetes and treatment with gangliosides on cardiac sympathetic innervation: a biochemical and functional study in mice. AB - The effect of long-term diabetes on cardiac sympathetic innervation was investigated in genetically obese diabetic mice (db/db). Previous studies have shown the presence of a peripheral neuropathy starting a few months after birth, and we recently reported a significant reduction of myocardial norepinephrine (NE) levels in the hearts of diabetic mice at the age of 6 months. In the present study, histofluorescence analysis of comparable sections of cardiac tissue of both control and diabetic animals confirmed the picture of a sympathetic denervated heart in this experimental model. Furthermore, functional studies in isolated atria revealed a difference between the two groups of animals: in fact heart rate increases induced by transmural stimulation were significantly lower in diabetic mice. Since a bovine brain ganglioside mixture (Cronassial) has been extensively studied for its effect on peripheral diabetic neuropathy, a group of diabetic mice was treated throughout the sixth month with this drug (10 mg/kg/day i.p.). The ganglioside treated animals showed a marked recovery of atrial function and cardiac NE concentration. The above results clearly indicate sympathetic neural damage in db/db animals, likely related to an autonomic diabetic neuropathy and a possible protection by ganglioside of adrenergic nerves from this alteration. PMID- 2968356 TI - Early diagnosis of diabetic small fiber neuropathy by disturbed cold perception. AB - In diabetes mellitus, polyneuropathy is an important complication and should be diagnosed as early as possible in order to prevent damage to the patient. Determination of warm, cold, and heat pain thresholds enables one to judge small nerve fiber sensitivity. This investigation was carried out to determine which parameters best predict such alterations. Using a "Marstock" Thermostimulator, 26 diabetics and 32 healthy subjects were stimulated behind both medial malleoli. At three different rates of temperature rise, repeated warm, cold, and heat pain threshold determinations were performed. The variability of intraindividual threshold ranges was noted. While heat pain determinations were not useful, determination of cold perception, at a moderate rate of temperature change, proved to be the most reliable indicator of small fiber lesions. Cold thresholds as well as their intraindividual ranges were most often impaired. The importance of this clinical investigatory method is discussed with respect to the importance of early prophylaxis of complications such as trophic lesions. PMID- 2968357 TI - The objective assessment of visual contrast sensitivity by pattern reversal visual evoked potentials in diabetes. AB - Recent studies have described abnormalities of visual evoked potentials and pattern electroretinography in diabetics without retinopathy. The visual contrast sensitivity, determined by psychophysical tests, has proved to be abnormal in diabetic patients with and without clinical retinopathy. In this study we evaluated contrast sensitivity function using both electrophysiologic and psychophysical methods. The objective assessment of functional visual contrast sensitivity was superior to psychophysical evaluation in the detection of contrast sensitivity alterations. No relationships were found between contrast sensitivity dysfunction and abnormalities of pattern electroretinography or fluorescein angiography. Our data suggest that functional visual deficits might precede background retinopathy and that the involvement of foveal function is early and very frequent in diabetic patients, even if they have normal visual acuity. PMID- 2968358 TI - Prevalence of microangiopathic complications in hyperglycemia secondary to pancreatic disease. AB - Diabetes secondary to pancreatic disease (PD) represents a useful model for the study of the effects of chronic hyperglycemia on microangiopathic complications in the absence of those genetic factors predisposing to Type I diabetes. Our aim was to evaluate the prevalence of nephropathy and retinopathy in a group of 86 patients with PD. The genetic pattern, assessed by the determination of HLA antigens, was different than in patients with Type I diabetes. A family history of diabetes was present in 53% of the patients. The prevalence of retinopathy was 37%. Eighteen percent of the patients with duration of diabetes less than 10 years showed an albumin excretion rate (AER) greater than 40 mg/24 hr. The prevalence of pathologic microalbuminuria (greater than 40 mg/24 hr) was found in 29% of the patients with duration of diabetes greater than 10 years. The prevalence of pathologic microalbuminuria is related to the duration of diabetes. Both diastolic and systolic blood pressure is positively correlated to albumin excretion rate (p less than 0.02), suggesting a possible role of hypertension in the evolution of nephropathy. Sixty-one percent of the patients with AER greater than 40 mg/24 h had retinopathy, thus confirming the close association between renal and ocular complications. Abnormal microalbuminuria and retinopathy were not influenced by a family history of diabetes. We conclude that the prevalence of microangiopathic complications is similar to that seen in Type I diabetes, and the metabolic abnormalities of diabetes can play a direct role in the development of diabetic microangiopathy. PMID- 2968359 TI - Glomerular filtration rate in early experimental diabetes. AB - A noninvasive single injection technique for the measurement of glomerular filtration rate (GFR) using technetium99m diethylene triamine pentaacetic acid (DTPA) was developed for use in the rat. GFR measurements obtained by the technique correlated well with those obtained by Cr51 EDTA infusion (R = 0.95, n = 7). The coefficient of variation was 8.4%. GFR was measured over 4 weeks in diabetic and control rats. GFR increased with time in both groups, with no difference between the groups; however, when corrected for body weight, diabetes was associated with an increased GFR (diabetic 13.6 +/- 1.7 vs. control 10.4 +/- 0.1 ml/min/kg p less than 0.001). Insulin treated rats had higher GFRs than untreated diabetics (p less than 0.05), but GFR/kg was reduced to that of nondiabetic controls. High protein intake in diabetic rats caused an increase in GFR after 1 week of diabetes, but this was not sustained by the fourth week. Genetic hypertension and angiotensin converting enzyme (ACE) inhibition with ramipril had no effect on GFR in diabetic rats. We conclude that serial measurement of GFR in the diabetic rat is accurate and reproducible. Genetic hypertension, high protein intake, and ACE inhibition have little effect on GFR in experimental diabetes. PMID- 2968360 TI - Column liquid chromatographic separation of collagen alpha-chain polymers on sepiolite. PMID- 2968361 TI - Assignment of the configurations of the amino acids in peptidic siderophores. AB - Pyoverdins and azotobactins contain beta-hydroxyaspartic acid, N delta hydroxyornithine, citrulline and homoserine, in addition to the common protein amino acids. Configuration assignment of all of these was achieved by acid hydrolysis of the peptide, derivatization of the constituent amino acids to the N pentafluoropropionyl amino acid esters and gas chromatographic separation of the stereoisomers on capillaries coated with Chirasil-Val. This approach is straightforward for the protein amino acids, but the less common amino acids are either partially degraded during acid hydrolysis or their derivatives exhibit unfavourable gas chromatographic properties. By judicious combination of partial and total hydrolysis and dual derivatization, these problems may be overcome. PMID- 2968362 TI - Alterations in the T-lymphocyte subpopulation in patients with rhinoscleroma. AB - T-lymphocyte subpopulations were studied in a group of patients with rhinoscleroma due to Klebsiella rhinoscleromatis. The data demonstrated that these patients had a significantly greater number of T-suppressor/cytotoxic lymphocytes than did clinically healthy individuals. This finding correlated with a diminished response to the T-cell mitogen concanavalin A. The evidence indicated that the T-cell response of these patients was decreased and may reflect the host's response to the bacterial invader, thus explaining the chronicity of the disease. PMID- 2968363 TI - Serial immune evaluation of cyclosporine- and placebo-treated multiple sclerosis patients. AB - During an ongoing clinical trial of cyclosporine (CsA) immunosuppression therapy for chronic progressive multiple sclerosis (MS), a comparison was made of the immune responses of 18 CsA- and 18 placebo (P)-treated MS patients. Patients randomized to receive either CsA or P had identical entry immune profiles. However, these MS patients displayed significantly increased T-helper:T suppressor (TH:TS) ratios (P less than 0.01), percentage (%) active-T (P less than 0.01), % Ia+-T (P less than 0.05) and % Ta1+-T (P less than 0.01) cell phenotypes when compared to age-matched normal controls. Further, the MS-P treated patients displayed significant increases (all P less than 0.01) in % pan T, % helper-T, % active-T and % Ta1+-T cell phenotypes as well as panel mixed lymphocyte culture (panel MLC) functional responsiveness from entry to cumulative 12-month study data. In contrast, the MS-CsA-treated patients only displayed an increased % pan-T cell phenotype. The cumulative 12-month follow-up data showed that the MS-P-treated patients displayed significantly higher immune parameters than the MS-CsA-treated patients for % pan-T (P less than 0.05), % helper-T (P less than 0.01), TH:TS ratio (P less than 0.05), % active-T (P less than 0.01), % Ta1+-T cells (P less than 0.01) and panel MLC stimulation index (P less than 0.01). Thus, MS-CsA-treated patients did not display the progressive immune activation seen on serial evaluation during the follow-up time period that characterized the placebo-treated MS group. PMID- 2968364 TI - The U2 small nuclear ribonucleoprotein particle as an autoantigen. Analysis with sera from patients with overlap syndromes. AB - We identified eight patients whose sera contained autoantibodies to the U2 small nuclear ribonucleoprotein (snRNP), an RNA protein particle involved in the splicing of newly transcribed messenger RNA. Each of these patients had an overlap syndrome that included features of either systemic lupus erythematosus (SLE), scleroderma, and/or polymyositis. We then used these sera to characterize the autoantigenic polypeptides of the U1 and U2 snRNP particles. In immunoblots, all sera contained antibodies to the B" polypeptide of the U2 snRNP. A subset of these sera that more effectively immunoprecipitated the native U2 particle contained an additional antibody system that recognized the A' polypeptide of this snRNP. Antibodies eluted from the B" protein bound the A polypeptide of the U1 snRNP and vice versa. Moreover, antibodies to the B" polypeptide were accompanied by antibodies to the 68K and C polypeptides of the U1 snRNP. Finally, the A' and B" polypeptides remained physically associated after the U2 particle was cleaved with RNase. Thus these sera contain multiple autoantibody systems that, at one level, target two physically associated antigenic polypeptides of the U2 particle and, at another, target two snRNP particles which are associated during the splicing of premessenger RNA. These linked autoantibody sets provide further evidence that intact macromolecular structures are targeted by the immune response in SLE and related diseases. PMID- 2968365 TI - Modulating role for thromboxane in the tubuloglomerular feedback response in the rat. AB - Some studies have indicated that PGs can modulate the single nephron tubuloglomerular feedback (TGF) response. The aim of this study was to define the specific role of the vasoconstrictor PG, TX, by administration to rats of either vehicle (group 1; n = 20) or drugs that inhibit either cyclooxygenase (indomethacin [indo], 5 mg.kg-1, group 2, n = 17), TX synthetase (UK-38,485 [UK], 100 mg.kg-1, group 3, n = 19), or TX receptors (SQ-29,548 [SQ], 8 mg.kg-1, group 4, n = 14, or L-641,953 [L], 50 mg.kg-1, group 5, n = 8). Indo reduced excretion of the prostacyclin derivative 6-keto-PGF1 alpha and TXB2 and lowered whole kidney GFR and renal plasma flow, whereas UK lowered excretion of TXB2 only and did not change basal renal hemodynamics. The TGF response (assessed from reduction in proximal tubule stop-flow pressure (Psf, mmHg) during increases in perfusion of the loop of Henle (LH) from 0 to 40 nl.min-1) was unchanged after vehicle (9.8 +/- 0.5-10.9 +/- 1.0, NS) but blunted (P less than 0.001) by 40-65% in rats of groups 2-5 (indo, 11.1 +/- 1.0-4.4 +/- 0.7; UK, 9.0 +/- 0.8-4.8 +/- 0.7; SQ, 10.3 +/- 0.6-4.8 +/- 0.6; L, 10.7 +/- 0.5-6.7 +/- 1.3). This blunting was due to lower values for Psf at zero LH flow after indo, SQ, and L, and higher values of Psf at 40 nl.min-1 LH flow after indo and UK. The fall in single nephron GFR (SNGFR, nl.min-1) with increasing LH perfusion was unchanged after vehicle (10.9 +/- 2.8-11.2 +/- 0.8) but was blunted (P less than 0.05) by 45-55% in rats given indo (13.9 +/- 1.2-6.2 +/- 2.2) or UK (12.8 +/- 2.1-7.0 +/- 1.5). UK produced dose-dependent reductions in TXB2 excretion (IC50, 15 mg.kg-1) and inhibition of the TGF response (IC50: 30 mg.kg-1). After blockade of TX receptors by SQ, UK had no further affect on the TGF response. The fall in Psf at high LH flow was blunted (P less than 0.05) by indo and UK, whereas the rise in Psf at zero LH flow was blunted by indo, SQ, and L. In conclusion, endogenous TX generation can modulate the reductions in Psf and SNGFR during increased delivery of NaCl to the LH. PMID- 2968367 TI - Specific correction of impaired acid hydrolase secretion in storage pool deficient platelets by adenosine diphosphate. AB - Storage pool-deficient (SPD) platelets, which have decreased amounts of dense granule and/or alpha-granule constituents, contain normal amounts of lysosomal acid hydrolases, but in some cases exhibit impaired secretion of these enzymes. We examined this impaired secretion response in SPD patients with varying extents of granule deficiencies, and determined the effects of added dense-granule constituents. Acid hydrolase secretion was impaired in patients with severe dense granule deficiencies, but not in patients with lesser dense-granule deficiencies, including those with alpha-granule deficiencies as well. When dense-granule constituents (ADP, ATP, serotonin, Ca+2, pyrophosphate) were added to gel filtered platelets, ADP, but none of the other constituents, completely corrected the impairment of thrombin and A23187-induced secretion in SPD platelets. The concentration of ADP required to normalize thrombin-induced secretion varied markedly, from 0.01 to 10 microM, among the individual patients. Fixation of platelets with formaldehyde before centrifugation did not prevent the enhancement of secretion by ADP. Excess ATP, which acts as a specific antagonist of ADP mediated responses, completely blocked this enhancement of secretion in SPD platelets by ADP, and partially inhibited acid hydrolase secretion induced by low, but not high, concentrations of thrombin in normal platelets as well. Treatment of normal platelets with acetylsalicylic acid in vivo, but not in vitro, produced an impairment of acid hydrolase secretion similar in extent to that in SPD platelets, but which could not be completely corrected by added ADP. One possible explanation of these results is that the impairment of acid hydrolase secretion may be secondary to the dense-granule deficiency in SPD platelets, and that secreted ADP may potentiate the lysosomal secretion response in normal platelets as well. PMID- 2968366 TI - Effect of age on myocardial adaptation to volume overload in the rat. AB - To test the hypothesis that the capacity for left ventricular (LV) adaptation to volume overload might diminish with age, we examined the hemodynamics and degree of myocardial hypertrophy in response to aortic insufficiency in young adult (9 mo) and old (18 or 22 mo) Fischer rats. Before, immediately after, and at 2 and 4 wk after creating aortic insufficiency, LV and aortic pressures were measured using a catheterization technique. 4 wk after surgery, we measured aortic flow, and estimated the LV passive pressure-volume relationship and the degree of LV hypertrophy after killing. Immediately after the surgical creation of aortic insufficiency, both young and old rats showed similar elevation of LV end diastolic pressure (from 4.8 +/- 0.6 to 12.0 +/- 1.5 mmHg in the young rats, P less than 0.01; from 4.9 +/- 0.4 to 11.0 +/- 0.7 mmHg in the old rats, P less than 0.01). In the young rats LV, end-diastolic pressure decreased to 8.0 +/- 1.0 and to 8.5 +/- 0.9 mmHg at 2 and 4 wk (P less than 0.05). In contrast, LV end diastolic pressure at 2 (16.9 +/- 3.1 mmHg) and 4 wk (16.1 +/- 2.7 mmHg) in the old rats was even higher, compared with the values measured immediately after aortic insufficiency. At 4 wk, LV end-diastolic meridional wall stress (calculated from the in vivo LV end-diastolic pressure, and the pressure-volume relationship and muscle mass obtained after killing) was higher in the old rats than in the young rats. In the young rats, the diastolic pressure-volume relationship at 4 wk shifted to the right (P less than 0.01), and LV dry weight, LV dry weight/tibial length, and protein content of the LV myocardium increased by 26% (P less than 0.01), 24% (P less than 0.01), and 33% (P less than 0.01), respectively. However, old rats with aortic insufficiency did not show a significant change in the pressure-volume relationship, dry weight, or protein content at 4 wk. These results suggest that advanced age diminishes the capacity for LV hypertrophy in response to a volume overload, and this reduced LV hypertrophic response in the old rats resulted in persistent elevation of LV end diastolic pressure and wall stress. PMID- 2968370 TI - Attributional style of depressed chronic low back patients. AB - This study used the Attributional Style Questionnaire to study the attributional styles of depressed and nondepressed chronic low back pain patients (N = 91) in order to test the Revised Learned Helplessness model's prediction of differences between the two. The results partly supported the hypothesis; an internal, stable, global style for negative events distinguished the depressed group from the nondepressed, but there were no differences in attributional style for positive events. The findings are consistent with recent reviews of the literature that have reported general support for the negative outcome style, but consistent failure to confirm the predictions associated with positive outcome style. In addition, the attributional style was not common to all subjects in the depressed group, which suggested that other factors may be involved in the development of different subtypes of depression. Implications for studying attributional aspects of depression and chronic low back pain are discussed. PMID- 2968368 TI - Synthesis of atrial natriuretic polypeptide in human failing hearts. Evidence for altered processing of atrial natriuretic polypeptide precursor and augmented synthesis of beta-human ANP. AB - To elucidate the synthesis of atrial natriuretic polypeptide (ANP) in the failing heart, 20 human right auricles obtained at cardiovascular surgery were studied. The concentration of alpha-human ANP-like immunoreactivity (alpha-hANP-LI) in human right auricles ranged from 13.8 to 593.5 micrograms/g, and the tissue alpha hANP-LI concentration in severe congestive heart failure (CHF) (New York Heart Association [NYHA] functional class III and class IV) (235.4 +/- 57.2 micrograms/g) was much higher than that in mild CHF (NYHA class I and class II) (52.5 +/- 15.6 micrograms/g). Atrial alpha-hANP-LI levels were significantly correlated with plasma concentrations of alpha-hANP-LI in these patients (r = 0.84, P less than 0.01). High performance gel permeation chromatography and reverse phase high performance liquid chromatography coupled with radioimmunoassay for ANP revealed that the alpha-hANP-LI in the human auricle consisted of three major components of ANP, gamma-human ANP (gamma-hANP), beta human ANP (beta-hANP) and alpha-human ANP (alpha-hANP). Comparing percentages of gamma-hANP, beta-hANP, and alpha-hANP in alpha-hANP-LI in severe CHF with those in mild CHF, the predominant component of alpha-hANP-LI was gamma-hANP in mild CHF, whereas beta-hANP and/or alpha-hANP were prevailing in severe CHF and, especially, beta-hANP was markedly increased in human failing hearts. These results demonstrate that the total ANP concentration in the atrium of the human heart is increased in severe CHF and that the increase of ANP in the human failing heart is mainly due to the increase of small molecular weight forms of ANP, beta-hANP, and alpha-hANP, especially beta-hANP, and indicate that the processing of ANP precursor, or gamma-hANP, in the human failing heart differs from that in the normal heart, suggesting that the failing heart augments synthesis and secretion of ANP as one of its own compensatory responses. PMID- 2968369 TI - Nonbinding inhibitory antiinsulin receptor antibodies. A new type of autoantibodies in human diabetes. AB - Sera and their IgG from 10/104 diabetic patients (five with insulin-dependent and five with noninsulin-dependent diabetes, NIDDM), contained antibodies that bound 125I-labeled purified human insulin receptors. 9 of these 10 sera failed to inhibit insulin binding (to rat hepatocytes and human placental membranes), did not stimulate glucose oxidation (by isolated rat adipocytes), and did not bind human placental IGF-1 receptors. Only one serum (and its IgG) modestly inhibited insulin binding and stimulated glucose oxidation. We conclude (a) that sera from 9/104 diabetics (five insulin-dependent and four noninsulin-dependent) contained a newly identified species of IgG antiinsulin receptor autoantibodies (AIRA), which bound to the insulin receptor at a locus different from the insulin binding site and did not inhibit insulin binding; and (b) that only 1/104 diabetic sera contained low-titer "conventional" antiinsulin receptor autoantibodies that bound to the insulin receptor at or near the insulin binding site, inhibited insulin binding and caused a clinical condition, which was difficult to distinguish from typical NIDDM. PMID- 2968371 TI - A review of criterion validation research on the Questionnaire on Resources and Stress for Families with Chronically Ill or Handicapped Members. AB - The Questionnaire on Resources and Stress for Families with Chronically Ill or Handicapped Members (QRS) has been used as a research instrument in studies that compared parents of clinical groups with normal controls, parents of children with different clinical conditions, and pre- and postinterventions. These studies are analyzed in terms of the relationship of 15 QRS scales to child variables (e.g., age, degree of handicap); parent variables (e.g., marital status, educational level); and family variables (e.g., nationality/culture). It was concluded that the QRS fulfills four requirements for an acceptable level of validity: (1) the relationship between predictor and criterion scores is statistically significant; (2) use of the test probably results in more correct decisions than would be made by resorting to base rates alone; (3) the test possesses some utility; it would result in some gain to the user; and (4) the test provides some unique information. PMID- 2968372 TI - Peripheral T lymphocyte subsets in rapidly progressive periodontitis. AB - The peripheral blood lymphocyte subsets of 10 patients with rapidly progressive periodontitis were investigated with monoclonal antibody. In 4 patients, the T helper/T suppressor ratio was increased. 5 others had a slightly reduced T helper/T suppressor ratio as compared to control group. These findings may indicate a possible cellular immune response in the pathogenesis of rapidly progressive periodontitis. PMID- 2968373 TI - Photopatch testing: a six-year experience. AB - The results of photopatch testing performed at the Mayo Clinic from 1980 through 1985 were analyzed. During the 6 years, 70 patients underwent photopatch testing. Of the 70 patients, 27 (38.5%) had positive photopatch test reactions and 22 (31%) had ordinary contact reactions. Of the 27 patients with photocontact reactions, 14 had relevant reactions. The most frequent positive reactions were to chlorpromazine (13 patients), musk ambrette (nine patients), and promethazine (eight patients). Photopatch testing is an essential part of the evaluation of patients who are suspected of having photosensitive dermatitis. In some patients with recalcitrant eruptions, testing may help in excluding photosensitivity as a possible contributing factor. PMID- 2968374 TI - Antipsoriatic, erythematogenic, and Langerhans cell marker depleting effect of bath-psoralens plus ultraviolet A treatment. Comparison of 8-methoxypsoralen and trimethylpsoralen photosensitization. AB - Successful results have been reported for bath-psoralens plus ultraviolet A (PUVA) treatment of psoriasis with the use of either a low-sensitizing psoralen (methoxsalen) or a high-sensitizing psoralen (trioxsalen), but no evidence has been presented that phototoxicity would be a prerequisite for antipsoriatic activity of these treatments. To study therapeutic-to-phototoxic ratios, bath PUVA was applied to psoriasis patients with the use of either a 0.2 mg/L solution of trioxsalen or a 0.4 mg/L solution of methoxsalen. The average minimal phototoxic ultraviolet A (UVA) dose (MPD) obtained after 15 minutes' bathing was 0.86 joule/cm2 for trioxsalen and 9.76 joules/cm2 for methoxsalen. In the first part of the study phototoxically equipotent treatment schedules were used, compensating the much lower phototoxic potential of methoxsalen by using about 10 times larger UVA doses compared to the doses used with trioxsalen. A healing rate of 71% +/- 25% was recorded for trioxsalen and 63% +/- 34% for methoxsalen treatment (mean +/- SD). Both PUVA treatments decreased epidermal Langerhans cell counts to 10% to 20% of the control value. In the second part of the study, the much lower phototoxic potential of methoxsalen was not compensated for. Instead, methoxsalen bath-PUVA was carried out with the use of UVA doses physically similar to those used in the trioxsalen therapy series. Surprisingly enough, even this very suberythemal methoxsalen therapy caused a significant healing effect (51% +/- 10%). Langerhans cell depletion in methoxsalen-bathed areas (reduction to 53% of the control value) was much less than that caused in trioxsalen-bathed areas (reduction to 11% of the control value).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968375 TI - Acne necroticans (varioliformis) and Staphylococcus aureus. PMID- 2968376 TI - Peripheral edema and oral acyclovir. PMID- 2968377 TI - Long-term radiographic follow-up after isotretinoin therapy. AB - We evaluated the effects of long- and short-term isotretinoin therapy on the skeletons of patients. Eight patients who were treated with isotretinoin for disorders of keratinization received frequent radiographic evaluations for 4 to 9 years. Seven patients developed multiple hyperostoses at the spine and extremities. Hyperostoses increased in size and number over the course of therapy, although relatively few sites were symptomatic. Hyperostoses typically developed first in the spine and later in the extremities, where both bilaterally symmetric and asymmetric involvement was observed. After 5 years of therapy one patient did not develop hyperostosis. In a group of nine patients who received a relatively high dose of isotretinoin in 1982 for the treatment of acne, two patients developed tiny, asymptomatic hyperostoses. One patient had hyperostoses 1 year after isotretinoin therapy, which remained unchanged 3 years later, whereas the other patient had one hyperostosis 4 years after therapy had been stopped. Although we suspect that these hyperostoses were retinoid induced, they should not be of concern for the patient needing routine isotretinoin therapy for the treatment of cystic acne. PMID- 2968378 TI - AIDS-associated Kaposi's sarcoma: variables associated with survival. AB - The records of 187 consecutive patients with acquired immunodeficiency syndrome (AIDS)-associated Kaposi's sarcoma were analyzed retrospectively for a number of prognostic variables. In a multivariate analysis, the initial site of disease was found to be related to survival. Initial lesions on the skin of the lower extremities or in the lymph nodes were associated with longer survival (p = 0.005 and p = 0.01, respectively). Higher helper/suppressor T cell ratios were strongly associated with longer survival (p less than 0.0001). Age and serum IgG antibody levels to cytomegalovirus did not appear to correlate. These results suggest that there are different subgroups of patients with AIDS-associated Kaposi's sarcoma, and that the initial site of disease, as well as immunologic parameters, may be useful in prognosis. PMID- 2968379 TI - The red man syndrome. Exfoliative dermatitis of unknown etiology: a description and follow-up of 38 patients. AB - Thirty-eight patients with erythroderma of unknown etiology were diagnosed over a 15-year period, and represented 19% of all patients admitted to our department for erythroderma. The male:female ratio was 6.6:1, and the median disease duration was 2 years (range 1 to 23 years). Keratoderma of palms and/or soles was seen in 79%. Laboratory findings were normal, except for an increased IgE level in 69% of the patients studied. Lymph node histology showed dermatopathic lymphadenopathy. Bone marrow investigation results were normal in 48%, or showed eosinophilia (32%) or hyperplasia (20%). Initial skin biopsies showed nonspecific histology in most patients, but later biopsies revealed pleomorphic infiltration. During the observation period four patients progressed to mycosis fungoides and another nine patients were suspected of having mycosis. None developed Sezary's syndrome. Only one third of the patients went into complete remission; half of them died during the observation period. Patients with erythroderma of unknown etiology are predominantly men and seem to belong to a certain subgroup--herein called the red man syndrome. PMID- 2968380 TI - Linear seborrheic keratoses associated with underlying malignancy. AB - A clinical syndrome, consisting of eruptive linear seborrheic keratoses associated with adenomatous colonic polyps (malignant but without invasion of the stalk) and/or frankly invasive colonic adenocarcinomas, is described in five patients. Partial to almost complete regression of the cutaneous lesions over the succeeding months to years was noted after removal of the internal malignant tumors in all five patients. Resolving lesions were characterized by an accentuation of chronic inflammatory infiltrate both within the epidermis and dermis, suggesting perhaps that the regression of the skin lesions may be related to enhanced immunologic response concomitant with the removal of the underlying malignancy. PMID- 2968381 TI - Granulomatous rosacea. PMID- 2968382 TI - Glass ionomer-silver cermet Class II tunnel-restorations for primary molars. AB - Tunnel preparations preserve the anatomical marginal ridge and minimize the loss of healthy tooth structure adjacent to the carious lesion. When the practitioner has developed proficiency in restoring class II carious lesions with tunnel restorations, less treatment time is required than with traditional class II preparations. The technique for restoring a primary first molar with a class II carious lesion, using a tunnel preparation and Ketac-Silver restorative material is described. PMID- 2968383 TI - Prevalence of bipolar disorder in a psychogeriatric population. AB - Bipolar affective disorder arising for the first time in old age (60 years and over) has not been extensively studied. The authors present a prevalence study of mania arising after age 60. Of 217 patients admitted to our unit in a 2-year period, 10 (4.7%) showed symptoms of mania, using the DSM-III criteria. This constitutes 9.3% of 108 affective disorder patients admitted during the same period. An important factor in the precipitation of these attacks was marital discord, leading, in several cases, to separation, even in this age group. PMID- 2968385 TI - Past loss as a symptom formation factor in depression. AB - We examined the relationship between past loss and endogenous subtyping of depressed patients. In a sample of 124 female depressed inpatients there was no association between past loss and RDC endogenous or DSM-III melancholic subtyping. We thus failed to replicate Brown and colleagues' finding that past loss is a symptom formation factor. PMID- 2968384 TI - Factors affecting help-seeking during depression in a community sample. AB - Little is known about factors that influence community residents to seek professional help while experiencing diagnosable episodes of depression. The present study utilized longitudinal data from 96 female subjects to examine whether clinical and psychosocial features of a recent depressive episode, as well as preexisting psychiatric and psychosocial characteristics, could distinguish between individuals who (a) did and did not seek help during their episode and (b) chose to consult one professional source rather than another. Results showed that less than half of the sample sought professional help. Few variables could distinguish subjects who sought help from those who did not. Instead, subjects consulting mental health specialists were more clinically impaired and had fewer psychosocial assets than both those consulting nonpsychiatric physicians and those seeking no help. Subjects in the latter two groups were indistinguishable from one another on the assessed variables. Results were cross-validated with a smaller sample of male community residents. PMID- 2968386 TI - Cognitive function in manics with associated neurologic factors. AB - Researchers studying cognition in mania have assumed that mania is a homogeneous entity. Recent preliminary evidence indicates that some manic syndromes may be preceded by medical, pharmacological, and neurologic antecedents. While DSM-III suggests that mild cognitive impairment may be associated with these manic syndromes, studies to date have not documented this assertion. We compared bipolar patients with antecedent neurologic factors (neurologic manics, NM) to bipolar patients without such histories (primary manics, PM) on standard neuropsychological measures and clinical parameters to ascertain whether cognitive testing could be used as an adjunctive diagnostic tool in defining this subgroup of patients. Results indicated that the NM group was more dysfunctional in intellectual functioning and course of psychiatric illness than the PM group. PMID- 2968388 TI - Cortisol nonsuppression in depression: relationship to clinical variables. AB - The dexamethasone suppression test was performed in 63 depressed patients and 43 normal controls. Cortisol nonsuppression was found in 47.6% of the depressed patients and 4.6% of the controls. Among depressed patients both age and reported weight loss, but not severity of depression, were significantly related to postdexamethasone plasma cortisol levels. Patients who were cortisol suppressors had experienced significantly more life events before the onset of depression and had significantly higher hostility scores on a personality questionnaire than cortisol nonsuppressors. PMID- 2968387 TI - A comparison of normal, bipolar and seasonal affective disorder subjects using the Seasonal Pattern Assessment Questionnaire. AB - The Seasonal Pattern Assessment Questionnaire is an instrument for retrospective self-rating of change in mood and vegetative functions with the seasons. It has been used in studies to identify and characterise patients with Seasonal Affective Disorder (SAD) in three countries. In this paper, the test-retest reliability of all items and the sensitivity to differences between groups have been investigated. Clear differences were found between the SAD group and a normal group. A group of bipolar affective disorder patients had intermediate scores on reported seasonal changes between the normal and SAD groups. All groups reported significant changes in mood and vegetative functions during the winter in the direction symptomatic for SAD suggesting that SAD patients may suffer an exaggerated form of a normal seasonal variation. PMID- 2968389 TI - Life history factors associated with neurotic symptomatology in a rural community sample of 40-49-year-old women. AB - Associations between a range of life history variables and neurotic pathology (measured by the Crown-Crisp Experiential Index) were examined in a rural community sample of 208 women aged 40-49. Significant childhood associations were a poor maternal relationship, parental discord, nervousness and not liking school. Significant later associations were a poor marital relationship and emotional and behavioural problems in children. There were significant intercorrelations between the significant childhood variables and between the later significant variables but not between the early and later variables. The predictor equations derived from multiple regression analysis fitted significantly a small replication patient sample. PMID- 2968390 TI - Premenstrual relapse of puerperal psychosis. AB - Eight patients suffering from puerperal psychosis rapidly recovered, then relapsed shortly before the onset of their first menstrual period; five of them had repeated premenstrual relapses. This clinical observation supports a hormonal aetiology for post-partum psychosis. PMID- 2968391 TI - Plasma HVA levels in depressed patients and controls. AB - Plasma levels of the dopamine metabolite homovanillic acid (HVA) were examined in depressed patients and controls. There were no significant differences between the groups. In particular, there were no significant differences for plasma HVA levels between depressed patients with and without melancholia or between depressed patients who were cortisol nonsuppressors or suppressors. PMID- 2968392 TI - A study of the age-related acceleration of glycation of tissue proteins in rats. AB - The relationship between age-related glycation and a fluorescent product attributable to the advanced Maillard reaction was investigated in the aortas of rats between 4 and 120 weeks of age. The early-stage product of the Maillard reaction was measured using furosine (epsilon-N-(2-furoylmethyl)-L-lysine) as a marker. The advanced product was measured by fluorescence high-performance liquid chromatography. The level of furosine in the aorta increased with aging to reach a maximum value in rats of 50 to 70 weeks of age, and decreased in rats from 90 to 120 weeks of age. By contrast, the level of the putative advanced product reached a maximum in 90-week-old rats and remained at the maximum level in 120 week-old rats. The level of glycated hemoglobin showed no significant change in rats of more than 14 weeks of age. The advanced Maillard products may play an important role in the pathophysiology of aging arteries. PMID- 2968393 TI - [Recourse to amniocentesis after the age of 38. Factors related to access to medical care]. AB - Antenatal screening by amniocentesis was not much used in Haute-Normandie (France) in 1981. This study compared 59 women aged 38 and over who did benefit from this test with a control sample of 71 women who delivered in the same region at the same time and who were not screened. The use of the screening was linked to the socio-professional category of the woman, to her professional activity, to her nationality, to where she studied, to her parity, and the quality of her obstetrical care. The information that the women who were screened had about trisomy 21 and its screening was far higher in those who were screened. Regression analysis indicates that there are 3 factors that play an important role: whether the specialist or the general practitioner offered the screening test and the socio-professional category of the woman. Amniocentesis could be used much more efficiently if the population was better informed and if the medical corps made a special effort as well as providing more screening facilities. PMID- 2968394 TI - [Assessment of the inflammatory evolution of the sequelae of salpingitis]. AB - Fifty-four patients who had consulted because of their sterility had a laparoscopy carried out with bacteriological, cytological and biochemical studies of the peritoneal fluid as well as a histological assessment of the peritoneal adhesions. The purpose of this prospective study was to show the usefulness of laparoscopy in pre-operative assessment of tubal sterility and to look for objective criteria of the progress of inflammation. To do the study, two groups of women in different clinical stages (29 patients with no pelvic infection and 25 patients who had salpingitis) were divided into three sub-groups: 25 free of disease, 4 subsiding salpingitis patients and 25 with tubo-peritoneal sequellae. Different samples taken with the laparoscope made it possible to decide whether the inflammatory process was carrying on or not in these 25 cases. Using swabs for bacteriological examinations we had to employ transport medium and this showed bacteria in 7 cases. We found a significant correlation between the serum and peritoneal levels of anti chlamydia antibodies and the levels of AC antibodies when there was a tubal lesion. There was little value in carrying out cytology on the peritoneal fluid except when the histology showed that this was necessary. The level of serum and peritoneal orosomucoid was found to be different in the two groups of patients. This study shows that it is necessary to assess several different parameters to exclude pelvic inflammatory disease before resorting to reparative tubal surgery. PMID- 2968395 TI - Alternative methods of communication for intubated patients in critical care. PMID- 2968396 TI - Immunoelectron microscopy of endothelial cells in rat incisor suggests that most basement membrane components are produced by young cells, whereas heparan sulfate proteoglycan is produced by both young and old cells. AB - When periodontal capillaries of rat incisor tooth were immunostained for four basement membrane components (laminin, collagen IV, fibronectin, heparan sulfate proteoglycan), all four were detected in the secretory organelles of endothelial cells located within 3 mm of the tooth's proximal end, but only the proteoglycan was observed in cells located 4 mm away and beyond (Experiment I). [3H]-Thymidine autoradiography revealed that the endothelial cells located at the tooth's proximal end were young and actively dividing, whereas those located 4 mm or more away were older and generally quiescent (Experiment II). Since immunostaining of a cell's secretory organelles for a given substance indicates production of this substance, the first experiment shows that endothelial cells at the proximal end produce the four basement membrane components. The second experiment discloses that these cells are young. As for the endothelial cells located 4 mm or more beyond the proximal end, the first experiment reveals that they produce only heparan sulfate proteoglycan, while the second shows that they are relatively old. Production of laminin, collagen IV, and fibronectin only by young cells implies that these substances are long-lived and stable components of basement membrane, whereas production of the proteoglycan by both young and old cells implies that it is labile and continually replaced. PMID- 2968397 TI - Plasma beta-thromboglobulin to platelet factor 4 ratios as indices of vascular complications in essential hypertension. AB - The ratio of the plasma level of beta-thromboglobulin (beta-TG) to platelet factor 4 (PF-4) which is regarded as a most reliable indicator of platelet activation in vivo, was followed in 52 subjects at various stages of essential hypertension according to the WHO classification. These comprised 30 cases at stage I, 19 cases at stage II and three cases at stage III, and 20 age-matched normotensive control subjects. The observed beta-TG:PF-4 ratio in the hypertensive patients was 4.59 +/- 0.20, which was significantly higher than the value of 3.13 +/- 0.19 recorded in the normotensive control subjects. According to the WHO classification, beta-TG:PF-4 ratios in hypertensive patients at stages I, II and III were 3.93 +/- 0.19, 5.31 +/- 0.35 and 6.56 +/- 0.12, respectively. The beta-TG:PF-4 ratio revealed a tendency of platelet activation to increase with advanced progress of hypertensive vascular lesions. These results suggest that the abnormal platelet function observed in patients with essential hypertension plays an important role in the development of hypertensive vascular complications. PMID- 2968398 TI - Generation of helper cell-independent cytotoxic T lymphocytes is dependent upon L3T4+ helper T cells. AB - The role of L3T4+ (CD4+) Th cells in generation of CTL specific for discrete minor histocompatibility Ag was investigated. Suppression of the function of Th cells in vivo by chronic treatment with anti-L3T4 mAb prevented congenic strains of mice from being primed and from generating CTL specific for Ag encoded by the minor histocompatibility loci--H-3, H-1, and B2m. Analysis of proliferative responses and lymphokine secretion of cells from animals primed with one of these minor H Ag, beta 2-microglobulin, but not treated with anti-L3T4 antibodies, indicated that L3T4- class I MHC-restricted T cells were themselves responsible for the very great majority of the observed minor H Ag-specific proliferation and secretion of lymphokines associated with both T cell proliferation and activation of CTL. All together, the data indicate that in responses against discrete minor H Ag, L3T4+Th-independent CTL are generated through an L3T4+Th-dependent pathway. PMID- 2968400 TI - Defective thymic education of L3T4+ T helper cell function in graft-vs-host mice. AB - Our study investigates the effect of a prior graft-vs-host (GVH) reaction on the subsequent ability of irradiated, bone marrow-re-populated mice to develop T cell function. The results indicate that such GVH-bone marrow transplanted (BMT) mice do not generate CTL responses to trinitrophenyl-modified syngeneic cells (TNP self), but do generate strong CTL activity to H-2 alloantigens. This selective deficiency in TNP-self CTL response potential appeared as early as 10 days after GVH, and required both L3T4+ and Lyt-2+ donor T cells. The in vitro addition of either soluble Th factors or L3T4-enriched spleen cells from normal mice circumvented the defect in the TNP-self response in GVH-BMT mice. These results indicate that T effector function was not defective, and instead suggest a Th defect. Cell depletion and antibody-blocking, as well as IL-2 production experiments, indicate that the Th defect was selective for L3T4+ Th population and not for Lyt-2+ Th population. This defect in L3T4 Th function is not accounted for by the approximate twofold reduction in L3T4 cell numbers in GVH BMT mice, because IL-2 production and CTL generation to L3T4-dependent Ag were at least eightfold below control levels. Rather, defective L3T4 Th function appears to be the consequence of a GVH-induced defect in thymic maturation because the defect was corrected in vivo by a neonatal parental thymus graft before irradiation and bone marrow transplantation. This system may be useful for elucidating the role of the thymus in the maturation of Th cells. Our findings raise the possibility that impaired development of T cell function occurring in marrow grafted patients who have undergone a GVH reaction could be partly due to a GVH-induced thymic defect. PMID- 2968399 TI - Suppression of B cell function by methotrexate and trimetrexate. Evidence for inhibition of purine biosynthesis as a major mechanism of action. AB - Methotrexate (MTX) is a widely used drug in the treatment of a variety of human neoplasms. Trimetrexate (TMQ) is a lipid-soluble quinazoline derivate of MTX that, unlike MTX, is not dependent upon membrane folate transport for cellular entry. A number of studies have demonstrated that MTX and, more recently, TMQ possess potent immunosuppressive properties. To examine the cellular events associated with the immunomodulatory effects of anti-folates on humoral immunity, a murine B cell maturation model was used. In vitro, MTX and TMQ reduced the number of antibody-forming cells to SRBC, as well as IgM production. B cells stimulated with anti-Ig demonstrated a dose-related suppression in [3H]UdR incorporation after addition of either drug, suggestive of a decrease in de novo DNA synthesis. B cell activation events preceding S phase were also suppressed by both anti-folates, as evidenced by inhibition of RNA synthesis. However, neither drug affected surface expression of Ia Ag nor inositol phosphate accumulation. Addition of TdR caused a slight non-significant increase in the antibody-forming cell response in the presence of 10(-7) M MTX. However, addition of hypoxanthine or adenine, but not guanine, resulted in complete restoration. Timed addition revealed that the ability of MTX to suppress antibody responses was diminished if added after 48 h of culture, similar to the reversal of this suppression mediated by hypoxanthine. Cell cycle analysis of LPS-stimulated B lymphocytes demonstrated that both drugs modulated events preceding, as well as during, the S phase. The present studies suggest that although drug-induced impairments in dTMP biosynthesis may be responsible for deficient lymphoid proliferation, anti-folate induced impairment in purine biosynthesis is a major mechanism in anti-folate induced suppression of humoral immunity. PMID- 2968401 TI - Possible role of neuraminidase in activated T cells in the recognition of allogeneic Ia. AB - In a primary MLR, predominant stimulators in spleen cells are adherent cells and not B cells, although B cells are one of the cell types expressing a large amount of Ia molecules. Our previous experiments showed that T cells treated with neuraminidase (Nase) responded to an allogeneic Ia on B cells. In our experiments, the relationship between the responsiveness to the allogeneic Ia molecules on B cells and Nase activity of T cells was examined. The results showed that T cells increased in Nase activity with the acquisition of the reactivity to Ia on B cells. T cells from normal mice increased in Nase activity after the incubation for 3 days or more in MLR, and these T cells responded to allogeneic Ia on B cells. However, T cells from mice genetically deficient in Nase responded poorly to the Ia on allogeneic B cells even after the incubation in MLR for 3 days. T cells incubated for 3 days in MLR decreased in electrophoretic mobility, indicating the decrease of net negative charge of the cells, and increased in their binding of peanut agglutinin which has been reported to bind to galactosyl residues exposed on T cell surface by removing sialic acids. These results suggest that Nase in T cells was activated by the cultivation in MLR for 3 days, and sialic acids of some molecules on T cell surface were removed by the enzyme and, in turn, T cells acquired the responsiveness to allogeneic B cells in a secondary MLR. Thus, Nase was suggested to play a regulatory role in the recognition of Ia molecules in T cells. PMID- 2968402 TI - Synergistic interaction between complement receptor type 2 and membrane IgM on B lymphocytes. AB - We sought biochemical evidence for a role of C receptors types 1 (CR1) and 2 (CR2) in B cell activation. A flow cytometer was used to measure the fluorescence of tonsillar cells that had been loaded with the calcium-dependent indicator indo 1, and cells were stimulated by cross-linking cell-bound DA4.4 anti-IgM, Yz-1 anti-CR1 or HB5 anti-CR2 with goat anti-mouse IgG. There was a direct dose response relationship between the proportion of cells having increased cytoplasmic free calcium concentration (Cai) after addition of second antibody and the amount of cell-bound Fab' DA4.4. In contrast, no rise in Cai was observed after cross-linking bound Yz-1 or HB5. To determine whether CR1 or CR2 could modify the increase in Cai induced by cross-linking membrane IgM, Cai was monitored after addition of second antibody to cells bearing combinations of either Yz-1 or HB5 with a limited amount of DA4.4. The combination of Yz-1 with DA4.4 yielded little or no further increase in the percentage of cells responding to cross-linking with elevated Cai compared with DA4.4 alone. However, the combination of HB5 with limited DA4.4 synergistically enhanced this response, resulting in stimulation that was equivalent to that obtained with optimal concentrations of DA4.4. The synergistic effect of CR2 was also observed with avidin as the cross-linking reagent for bound biotinylated HB5 and DA4.4, occurred in the presence of EGTA, and did not require T cells. Studies of the proliferation of B cell-enriched PBMC demonstrated that, whereas HB5 coupled to Sepharose alone induced little or no DNA synthesis, the combination of HB5 with limited DA4.4 on Sepharose induced a dose-related synergistic increase in the incorporation of [3H]thymidine. PMID- 2968403 TI - The involvement of human tumor necrosis factors-alpha and -beta in the mixed lymphocyte reaction. AB - The influence of recombinant human TNF-alpha and -beta (rHuTNF-alpha and -beta) in a human mixed lymphocyte reaction (MLR) was investigated. The addition of 1000 U/ml of either cytokine at the initiation of culture caused up to a sixfold increase in [3H]thymidine incorporation by responder cells. Furthermore, it was found that endogenous HuTNF-alpha is produced after allogeneic cell interaction and can be detected in the MLR supernatant within 1 h of culture initiation. The results also show that, in the absence of exogenous HuTNF-alpha, antibodies to rHuTNF-alpha can cause a significant inhibition of the MLR. These observations indicate the importance of TNF-alpha in allogeneic cell interaction and raise considerations for the use of antibodies, or other antagonists, to TNF-alpha as regulators of disease states associated with cell-mediated immune reactions. PMID- 2968404 TI - Synthesis and regulation of complement protein factor H in human skin fibroblasts. AB - The alternative pathway of C activation is Ag-independent and forms a first line of defense against infection before immune response. The C3 convertase, C3bBb, formed during activation of the alternative pathway is tightly regulated, with destabilization produced by factor H. Using metabolic labeling with [35S]methionine, immunoprecipitation, and SDS-PAGE, we demonstrated that human skin fibroblasts synthesized and secreted factor H protein. Two forms of the protein were identified, the approximately 160-kDa form seen more prominently in serum and a 45-kDa form that has also been identified in serum. The cells contained two forms of factor H mRNA, 4.4 and 1.8 kb. IFN-gamma increased factor H protein synthesis and mRNA content. No effect was observed with LPS. Neither HepG2 cells or human peripheral blood monocytes synthesized factor H protein or contained factor H mRNA. PMID- 2968405 TI - Stimulation of human neutrophil adhesive properties by adenine nucleotides. AB - Inasmuch as adenine nucleotides may be secreted by platelets during inflammation, we sought to determine whether ATP and related compounds could serve as stimuli of neutrophil (polymorphonuclear cells, PMN) activation as manifested by an increase in their adhesive properties. Exposure of isolated human PMN to ATP or its nonhydrolyzable analog, adenosine 5'-O-(3-thiotriphosphate) did indeed stimulate an increase in cellular adhesive function as assessed by an increase in the surface expression of the leukocyte adhesion-promoting glycoprotein, Mo1 (CD11b/CD18), the initiation of PMN aggregation, and (in the case of ATP) the attachment of increased numbers of albumin-coated polystyrene latex beads. However, this increase in PMN adhesive function was not accompanied by the generation of products of the respiratory burst. These in vitro data suggest the possible influence of secreted adenine nucleotides in promoting neutrophil adhesion-dependent interactions at inflammatory sites in vivo. PMID- 2968406 TI - Human IFN-gamma production is inhibited by a synthetic peptide homologous to retroviral envelope protein. AB - A synthetic 17 amino acid peptide (CKS-17) homologous to a highly conserved region of human and animal retroviral transmembrane proteins was investigated for its influence on the in vitro production of IFN-gamma from human peripheral mononuclear cells. The results showed that CKS-17 coupled to a carrier protein, BSA, inhibited production of IFN-gamma in a dose-dependent manner. Controls, consisting of BSA, which had undergone the coupling procedure or neurotensin coupled to BSA in an identical manner as CKS-17, showed no such inhibition. Reduction in IFN-gamma production could not be attributed to decreased viability of cells, delay of IFN-gamma production or to involvement of suppressor cells. Moreover, inhibition of IFN-gamma production was not related to the inhibition of DNA synthesis. The inhibition appeared to be a direct effect of CKS-17 on IFN gamma-producing cells. Kinetic studies revealed that this suppression occurred when CKS-17 was introduced to the culture concurrent with or within 48 h after introduction of IFN inducers. Preincubation experiments showed that the presence of CKS-17 in the culture medium was not necessary to exert its inhibitory effect. These results suggest that a portion of retroviral envelope proteins possess important immunomodulatory actions. PMID- 2968408 TI - Subsets of T lymphocytes in peripheral blood of patients with oral lichen planus. AB - The T lymphocyte subsets in the peripheral blood were studied in 26 patients with oral lichen planus and in 11 normal controls. Lymphocyte subsets were identified with monoclonal antibodies (Ortho Diagnostic System Inc. Raritan, New Jersey, USA) by an indirect immunofluorescent antibody method. This study shows no significant difference between patients with oral lichen planus and healthy controls. The conclusion is that local immunity, due to the breakdown of skin associated lymphoid tissue, not a generalized immunologic disorder, is considered to be involved in the pathogenesis of oral lichen planus. PMID- 2968407 TI - Production of colony-stimulating factor by tumor cells and the factor-mediated induction of suppressor cells. AB - Spleen mononuclear cells of C3H/HeN mice were cultivated with mitomycin C-treated tumor cells, X5563, MH134, MM48, MM46, and FM3A/R, all of which were of syngeneic origin, in a medium containing normal syngeneic mouse serum but not FCS. There was a proliferative response to X5563, MH134, and MM48, but not to the two other tumor cells, MM46 and FM3A/R. The responder spleen cells were found to be nonadherent cells with a phenotype of Thy-1-L3T4-Lyt2-Ig-Macl-, which were neither mature T and B cells nor mature macrophage/granulocytes. It was also found that the proliferation of these nonadherent no-marker cells was mediated by tumor cell-derived soluble factors but not by direct stimulation with tumor cells. The responsible factor was a molecule(s) with a Mr of 23 to 25 kDa, which had a CSF activity inducing granulocyte (G)-, macrophage (M)- and G + M-colonies in the bone marrow cells. Neutralization tests of this factor-induced proliferation of spleen cells revealed that a major part of the factor may be GM CSF or a molecule closely related to it. Incubation of spleen mononuclear cells with these GM-CSF-like tumor cell factors resulted in induction of myeloblastic/promyelocytic cells with a phenotype of Mac-1+2+Ia+ Thy-1-L3T4-Lyt2 Ig- in the spleen cell cultures, which could suppress mitogenic responses of the spleen cells to T and B cell mitogens. GM-CSF-like activity could also be detected in the serum of mice bearing X5563, MH134, and MM48, but not in those bearing MM46 and FM3A/R. Subcutaneous inoculation of C3H/HeN mice with these X5563, MH134, and MM48 tumor cells generated massive metastasis in the lung and lymph nodes, whereas MM46 and FM3A/R produced no macroscopic tumor cell metastasis. These results strongly suggest the possibility that in some tumor cell-host systems, a GM-CSF-like factor(s) produced constitutively by the tumor cells may play an important role in the development of tumor metastasis, mediating through suppression of lymphoid tissues of the host. PMID- 2968410 TI - From bedpan to...? PMID- 2968409 TI - Myonecrosis following deep pelvic hyperthermia. AB - A case report is presented in which severe myonecrosis of the inferior anterior abdominal wall followed treatment of a recurrent pelvic tumour with an annular phased array (AA) deep heating device. This unusual complication has not been observed at this institution in 107 patients previously treated with deep pelvic or abdominal hyperthermia, who had undergone a total of 368 hyperthermia sessions. The injury was confirmed radiographically and histopathologically, and was associated with serum enzymatic elevations characteristic of muscular injury. The implications for clinical trials employing the AA or similar devices are discussed. PMID- 2968411 TI - Effects of alcohol on the central nervous system: implications for the neuroscience nurse. PMID- 2968412 TI - Care of the patient with Parkinson's disease: surgical and nursing interventions. AB - Parkinson's disease (PD), occurring in one million people with 50,000 new cases each year, is the most prevalent degenerative neurological disorder. With symptoms occurring insidiously and gradually, PD typically strikes in the sixth decade of life. Parkinsonian signs and symptoms include the classic combination of resting tremor, rigidity, bradykinesia and postural instability. The impact of the disease varies among individuals. Since the experiences of patients with Parkinson's disease are so varied and complex, nurses must play a pivotal role in their care. The quality of life of individuals with PD has changed dramatically since the advent of levodopa (L-Dopa) and other anti-Parkinson drugs. Recently autotransplantation of the adrenal medulla into the caudate nucleus, an investigational surgical treatment for Parkinson's disease, has been developed. Success with this procedure has been reported in two case studies. Autotransplantation is performed in several institutions in the United States. In this paper, the acute nursing care of individuals who have undergone autotransplantation, as well as short-term effects of the procedure on the lives of two patients and their families will be examined. This article addresses traditional medical treatment, physiology of movement and autotransplantation in addition to two case studies. PMID- 2968413 TI - Memory impairments: forgetfulness versus amnesia. AB - Two varieties of memory disorders can be distinguished--those characterized by the phenomenon of forgetfulness and those characterized by the presence of an amnesic state. Although these two conditions may appear similar, they have different anatomical correlates and functional significance. States of forgetfulness and amnesia arise from different etiologies, have different prognoses and require different therapeutic regimes. The purpose of this article is to distinguish these two varieties of memory disorders and contrast them as to anatomical features, functional differences, etiologies, prognoses and therapeutic management regimes. This should assist the neuroscience nurse to better understand relevant nursing assessment features and plan appropriate therapeutic nursing interventions for the client. Teaching protocols for families and significant others as well as clients with memory impairments should also be enhanced. PMID- 2968414 TI - Pediatric craniofacial reconstruction: an overview of perioperative management. AB - Pediatric craniofacial defects as a result of congenital malformation, tumor, or trauma are devastating to both the child and the family. These patients require extensive education, preparation, careful monitoring and psychological support. With continuing advances in surgical technology and technique, the effectiveness and utilization of pediatric craniofacial surgery is increasing. As this type of surgery is currently available at relatively few institutions, it has created a new nursing experience. A multi-disciplinary team approach has evolved at Texas Children's Hospital which necessitates active involvement of nursing staff. In this article the multi-disciplinary approach and nursing care utilized in the perioperative management of the craniofacial pediatric surgery patient will be discussed. PMID- 2968415 TI - Stroke recovery: review of the literature and suggestions for future research. AB - Stroke kills more than 175,000 individuals annually and ranks second as a cause of long-term physical disability among Americans. Yet little is known about the experience of recovery in stroke survivors, particularly on a longitudinal basis. Regardless of the distressing impact of stroke on the individual, there is a paucity of literature on the stress of the stroke experience. Critique of recent research in stroke recovery is presented as well as the impact of stroke on the individual. Clinical ethnography as an alternative method of studying stroke recovery is described. PMID- 2968416 TI - Parental attitude and coping behaviors in families of children with epilepsy. AB - This study investigated the relationship between parental attitudes toward their children's epilepsy and parental coping patterns. Subjects were 27 parents whose children had epilepsy. Parental attitude was measured using the Fishbein Expectancy-Value Model, while coping was measured using the Coping Health Inventory for Parents. Significant positive correlations were found between parental attitude and the coping pattern of Maintaining Family Integration, Cooperation and Optimistic Definition of the Situation (r = .42, p less than .02). Parental attitude was also positively related to the coping pattern of Maintaining Social Support, Self-esteem and Psychological Stability (r = .32, p less than .05). Results suggest parents with positive attitudes toward their children's epilepsy use more positive coping behaviors than parents with less positive attitudes. PMID- 2968417 TI - Episodic dyscontrol syndrome and head injury: a case presentation. AB - Although the individual's baseline neurological status may appear normal, episodic dyscontrol syndrome presents with recurrent outbursts of rage. In some cases family may describe a history of personality traits reflective of minimal brain dysfunction, and the study by Elliot confirmed organic cerebral disorders in 94% of the sample. Due to difficulty of diagnosis and lack of reporting, as well as social factors, the prevalence of this syndrome may be underestimated. This article discusses the syndrome, reviews a case history and outlines the role of the neuroscience nurse. PMID- 2968419 TI - Determining interrater reliability of nurses' assessments of pupillary size and reaction. AB - Pupil changes provide vital information related to the cause and location, and at times, severity of disease processes which alter level of consciousness. Size and reactivity are sensitive to a variety of influences including local eye damage, functional state of brainstem, local systemic drugs, seizures and anoxia. Therefore, accurate measurement of pupillary size and reactivity is essential. The purposes of this study were to determine if pairs of nurses (1) achieved the same measurement of pupil sizes with and without an objective measure, and (2) selected the same descriptor for pupillary reaction to light. Sixty-eight pairs of nurses employed in adult and pediatric units of a large teaching hospital participated in the study. Interrater reliability of pupillary assessment of 136 nurses was determined. There was no significant difference in the reliability of nurses assessing pupillary size regardless of whether they used a penlight pupil gauge to measure or used observation without a pupil gauge. Agreement among the nurses on the pupillary size was high. Agreement on the pupillary reaction, however, was poor to good. PMID- 2968418 TI - A pubococcygeal exercise program for simple urinary stress incontinence: applicability to the female client with multiple sclerosis. AB - Five women with multiple sclerosis (MS) and simple urinary stress incontinence were informally evaluated and taught a program of pubococcygeal exercises to determine if inadvertent loss of urine during activities which increase intraabdominal pressure could be improved or eliminated. A correlation seemed to exist between the client's ability to experience vaginal contraction during sexual climax and elimination or improvement of simple urinary stress incontinence using a pubococcygeal exercise program. The purpose of this article is to encourage nurses to consider a pubococcygeal teaching program for treatment of simple urinary stress incontinence in female clients with MS who do experience vaginal contraction during sexual climax. PMID- 2968420 TI - Development of a stroke family support and education program. AB - An interdisciplinary stroke family support and education program can provide families with basic information about stroke and rehabilitation in addition to providing support to families as they adapt to the crisis of stroke. The theoretical basis for such a program is discussed in this article as well as program development, implementation and evaluation. Included are program goals, content outlines and a referral process. PMID- 2968421 TI - Fiberoptic "awake" nasal intubation: physiological and personal concepts. AB - Use of fiberoptics is gaining in popularity, particularly in neurosurgery. Advantages of use of FNI for the patient with neurological dysfunction include: minimized increases of ICP; decreased use of opiates; reduced manipulation of the spine; direct visualization of the respiratory tract. PMID- 2968422 TI - GI drugs: histamine antagonists, sucralfate and metoclopramide. PMID- 2968423 TI - Ankle-brachial index (ABI) and leg blood flow following epidural anaesthesia. AB - The influence of the severity of occlusive arterial disease, as evaluated from the ankle-brachial index (ABI) values, on blood flow and the distribution of blood flow within the lower limb following epidural block to the level of Th 4 to 6 was studied in 48 patients (ABI values ranging from 0.45-1.62). Leg (LBF) and foot (FBF) blood flows were measured with plethysmography and skin blood flow (SBF) was evaluated from laser-Doppler flowmetry and temperature recordings. The resting control LBF, FBF and SBF values were similar for all patients. Following the epidural block mean arterial blood pressure (MAP) decreased by about 20 mmHg. LBF, FBF and SBF all increased. There were significant correlations between the ABI values and the changes following the epidural block in LBF and FBF but not between ABI and SBF changes as evaluated from laser-Doppler flowmetry. PMID- 2968424 TI - Evaluation of a laser Doppler multiprobe for detecting skin microcirculatory disturbances in patients with obliterative arteriosclerosis. AB - Laser Doppler fluxmetry (LDf) is a noninvasive technique measuring blood cell flux, e.g. in human skin. By a new integrated probe the measuring volume has been increased sevenfold. Using this probe we have investigated dynamic reactions in the skin microcirculation of the big toe in nine patients with various degrees of obliterative arteriosclerosis and in ten healthy control subjects. When position was changed from the supine to the sitting a significant decrease in skin perfusion was seen in healthy subjects but not in the patients. The time to peak flow during reactive hyperemia after a three minute arterial occlusion was significantly (p less than 0.01) longer in the patients (117 +/- 81 s) than in the controls (24 +/- 6 s). We conclude that LDf, using this multiprobe, is of value for diagnosing patients with obliterative arteriosclerosis, as well as for studying dynamic reactions in the human skin microcirculation in health and disease. PMID- 2968425 TI - High concentrations of Lp(a) lipoprotein in serum are common among patients with abdominal aortic aneurysms. AB - The serum concentrations of Lp(a) lipoprotein was determined in 57 patients with abdominal aortic aneurysms (AAAs) and in 56 controls. Twentynine of the AAA patients also suffered from other cardiovascular diseases and eleven had brothers and sisters with AAAs. Lp(a) was significantly higher among the AAA-patients than among the controls (22.3 +/- 24.3 vs 12.6 +/- 20.4 mg/dl, p less than 0.01). The 28 AAA-patients without other cardiovascular diseases had a somewhat higher Lp(a) in serum than the other AAA-patients, but this difference was not statistically significant (26.4 +/- 28.4 vs 18.5 +/- 19.3 mg/dl). The 11 AAA-patients with AAAs in the family didn't differ from the other AAA-patients concerning Lp(a) in serum. Several genetic markers were also studied, and the seven Kell-positive AAA patients had a significantly lower Lp(a) in serum than the 47 Kell-negative AAA patients (7.0 +/- 10.3 vs 25.7 +/- 25.2 mg/dl, p less than 0.05). PMID- 2968426 TI - Experimental evaluation of small diameter synthetic arterial grafts. AB - Platelets labelled with Indium-111 were used to examine the in vivo thrombogenicity of different vascular grafts. The deposition of platelets in two partly different kinds of umbilical vein grafts, double velour Dacron grafts and double velour Dacron grafts with internal collagen was studied as a function of time. The grafts were inserted end to side in the femoral artery of pigs and then imaged for 120 minutes. Platelet distribution was also studied by in vitro static imaging. No difference was seen between the different umbilical vein grafts. The double velour grafts accumulated more platelets, the platelets mainly located in the midportion of the graft. The collagen-impregnated grafts occluded within 45 minutes. PMID- 2968428 TI - [Correlation between breast development and hormone profiles in puberal girls]. AB - In order to examine the relation between breast development and hormone levels, serum levels of hormones including LH, FSH, PRL, progesterone, 17 alpha OH progesterone, estradiol (E2), pregnenolone, pregnenolone-sulfate, 17 alpha OH pregnenolone, cortisol, dehydroepiandrosterone (DHA), dehydroepiandrosterone sulfate (DHA-S), testosterone and delta 4-androstenedione were measured by RIA in 162 puberal girls aged 9 to 17 years. Height and body weight were also measured. Stages of breast development were classified into B1 to B5 according to Tanner et al. Results were as follow; 1) Both body weight and height increased as the breast stage advanced during puberty. 2) The correlation with the concentration of hormone such as LH, FSH, E2, DHA, DHA-S and T, and breast development was noticed during the puberal stage before the onset of menarche. A significant positive correlation was observed between DHA-S, body weight and each stage of breast development before and after the onset of menarche. From the results obtained above, the intimate relationship between breast development and steroid concentrations especially the adrenal steroid DHA-S was suggested. PMID- 2968427 TI - Complement activation in atherosclerosis: effect of angiography and surgery. AB - Complement activation in the plasma of patients with severe atherosclerosis (arterial occlusive or aneurysmatic disease) was investigated in this study. The effects of angiography and reconstructive arterial surgery (RAS) were also assessed. Atherosclerosis was not found to be associated with systemic complement activation. Angiography resulted in high levels of C3 breakdown products. Surgery caused a significant degree of systemic complement activation in both patients with atherosclerosis and controls. Post-operative levels of C3 breakdown products were significantly higher in atheromatic patients than in controls, most likely due to the insertion of dacron arterial prostheses in the first group. PMID- 2968429 TI - [Changes in maternal plasma levels of C21 and C19 steroid hormones during pregnancy]. AB - To study the changes in circulating steroid levels during pregnancy, ten steroids which included Progesterone (P4), 16 alpha OH-Progesterone (16P4), free and conjugated Pregnenolone (P5), 16 alpha OH-Pregnenolone (16P5), Dehydroepiandrosterone (DHA) and 16 alpha OH-Dehydroepiandrosterone (16DHA) were measured simultaneously by RIA in 84 normal pregnant women from the 5th to 41st week of gestation. Ratios steroid levels were also calculated. The results were as follows; 1. P4 increased as gestation advanced and reached the maximum (126 +/ 18.24 ng/ml) in the 28th week. The concentration 16P4 increased sharply after the 32nd week and reached the maximum (16.3 +/- 1.67 ng/ml) in the 39th week. 2. Both free and conjugated P5 levels did not show an increasing trend as pregnancy progresses. 3. The concentration of free and conjugated 16P5 increased rapidly after the 30th week and reached the maximum at 39th week (free 2.5 +/- 0.3 ng/ml conjugated 42.4 +/- 5.1 ng/ml). 4. Both free and conjugated DHA decreased as pregnancy progressed. On the other hand, the concentration of free and conjugated 16DHA increased as pregnancy progressed. 5. The ratio of 16P4 to P4, 16P5 to P5 and 16DHA to DHA significantly increased after 30 weeks of gestation. PMID- 2968430 TI - Vibration white finger. AB - Nineteen consecutive patients claiming compensation for vibration white finger were reviewed. The cold provocation test was found to be of no value in confirming a diagnosis of vibration white finger. Small cysts were apparent on the radiographs in 61% of patients' wrists but none had developed significant degenerative changes of the wrist or digits. In this group of claimants, 63% had carpal tunnel syndrome on nerve conduction studies. PMID- 2968431 TI - Exercise and smoking habits in patients with and without low back and leg pain. PMID- 2968432 TI - Chiropractic management of spondylolisthesis with spondylolysis of the pars interarticularis: an example of the single-case study experimental design. AB - Case records permeating the chiropractic literature, although claiming success utilizing conservative therapies, often are founded on isolated circumstances rather than scientific data. A detailed examination of such reports reveals a void with respect to definitive and specific approaches for the diagnosis and clinical management of disorders synonymous with chiropractic clinical practice. At best, therefore, such reports are fraught with empiricism, illustrating only the experiences of individual clinicians. The underlying difficulty encountered in reporting information on purely didactic grounds is likely due to the absence of a mechanism by which improvement in biomechanical function may be precisely and adequately quantified. In direct contrast, controlled clinical trials, as in medical research, offer the luxury of statistical clarity as to the selection of one treatment regimen over another. Researchers have indicated that the single case study experimental design may be of value in chiropractic clinical practice, allowing for the formulation of deductive conclusions derived from each case. To facilitate the process, implementation of both retrospective and prospective aspects are proposed modifications to the general scheme. It is the purpose of this article to employ the concept of the single-case study experimental design, illustrating a condition commonly encountered in chiropractic clinical practice, that of spondylolisthesis. In so doing, we attempt to adhere to the prescribed format, while outlining both the retrospective and prospective aspects, commensurate with such a problem within the clinical setting. PMID- 2968433 TI - Medical economics: prescription study. PMID- 2968435 TI - Anti-interleukin 2 receptor monoclonal antibodies spare phenotypically distinct T suppressor cells in vivo and exert synergistic biological effects. AB - The therapeutic efficacies of ART-18, ART-65, and OX-39, mouse antibodies of IgG1 isotype recognizing distinct epitopes of the p55 beta chain of the rat IL-2-R molecule, were probed in LEW rat recipients of (LEW X BN)F1 heterotopic cardiac allografts (acute rejection in untreated hosts occurs within 8 d). A 10-d course with ART-18 prolongs graft survival to approximately 21 d (p less than 0.001). Therapy with ART-65, but not with OX-39, was effective (graft survival approximately 16 and 8 d, respectively). Anti-IL-2-R mAb treatment selectively spared T cells with donor-specific suppressor functions; the CD8+ (OX8+ W3/25-) fraction from ART-18-modified recipients, and primarily the CD4+ (W3/25+ OX8-) subset from ART-65-treated hosts conferred unresponsiveness to naive syngeneic rats after adoptive transfer, increasing test graft survival to approximately 16 and 45 d, respectively. Concomitant administration of ART-18 and ART-65 to recipient animals in relatively low doses exerted a strikingly synergistic effect, with 30% of the transplants surviving indefinitely and 50% undergoing late rejection over 50 d. These studies provide evidence that anti-IL-2-R mAbs selectively spare phenotypically distinct T cells with suppressor functions. The data also suggest that in vivo targeting of functionally different IL-2-R epitopes may produce synergistic biological effects. PMID- 2968436 TI - Intrathymic elimination of Mlsa-reactive (V beta 6+) cells during neonatal tolerance induction to Mlsa-encoded antigens. AB - The cellular basis of neonatally induced T cell tolerance has been investigated in a model system in which usage of a particular TCR V beta segment (V beta 6) is strongly correlated with reactivity to antigens encoded by the Mlsa genetic locus. Expression of V beta 6 by peripheral T cells was virtually abolished in BALB/c (H-2d, Mlsb) mice rendered neonatally tolerant to DBA/2 (H-2d, Mlsa) lymphoid cells, whereas control V beta 8-bearing T cells remained at near normal levels. Further analysis revealed that elimination of V beta 6+ T cells occurred in the thymus of neonatally tolerant mice and could not be explained by receptor modulation or T cell chimerism. These data thus support the clonal deletion model of tolerance induction. PMID- 2968434 TI - Function and regulation of a murine macrophage-specific IgG Fc receptor, Fc gamma R-alpha. AB - Ligand binding specificities of two cloned murine Fc gamma Rs (Fc gamma R-alpha, Fc gamma R-beta [9]) were determined by gene transfer into Fc gamma R negative cell lines. Both receptors were expressed as full-length molecules capable of IgG immune complex binding that was inhibitable by the mAb 2.4G2. The ligand binding profiles of these receptors were indistinguishable whereby both bound immune complexed mouse IgG1, IgG2a, and IgG2b, but not IgG3. Neither receptor could bind monomeric IgG2a, indicating these receptors to be low-affinity IgG Fc receptors. Accumulation of the Fc gamma R-alpha mRNA can be induced with murine IFN-gamma at a concentration of 200 U/ml in the macrophage-like cell lines RAW 264.7 and J774a. The time course for induction indicates that the mRNA accumulation is transient but does not return to the uninduced level even after 50 h of treatment. Fc gamma R-beta mRNA was not induced by IFN-gamma, rather its expression was down modulated in mouse peritoneal macrophages. Both RAW and J774a cells lines exhibited increased receptor levels after IFN-gamma stimulation as measured by 125I-2.4G2 and ligand binding. In the absence of IFN-gamma, the RAW and J774a cell lines were minimally phagocytic, while P388D1 cells were actively phagocytic. In the presence of IFN-gamma, however, RAW 264.7 and J774a cells were induced to become actively phagocytic. Induction of Fc gamma R-alpha mRNA and protein by IFN-gamma may be part of the process by which macrophages become activated to engulf antibody-coated particles. PMID- 2968437 TI - The physiological basis of slow locomotion in chamaeleons. AB - The African chamaeleon, Chamaeleo senegalensis, will not move faster than approximately 0.1 m/second at 23 degrees C, whereas the lizard Agama agama, like most lizards its size, runs at speeds more than 10X as fast. To account for this difference, we measured various physiological parameters of the iliofibularis muscle of both lizards. The maximum speed of tetanic contraction of unloaded Chamaeleo muscle was half as fast as that of Agama muscle (2.5 vs. 5.8 resting lengths per second). Heavily loaded Chamaeleo iliofibularis contracted at nearly 1/4 the speed of Agama muscle. Time to peak isometric twitch tension and time to half relaxation were twice as long in Chamaeleo as in Agama (122 vs. 58 msec, and 168 vs. 81 msec). Much more of the Chamaeleo muscle consisted of tonic muscle fibers, and the Chamaeleo muscle, compared to Agama muscle, showed physiological evidence of having a significant amount of tonic fibers (potassium contracture and high tetanus to twitch ratios). Finally, the myofibrillar ATPase activity of the Chamaeleo muscle was 1/3 that of Agama muscle. Thus, these results show that the slow locomotion of old world chamaeleons can, in part, be explained by the physiology, biochemistry, and fiber-type distribution of their muscles. PMID- 2968438 TI - Histochemical and biochemical characterization of two slow fiber types in decapod crustacean muscles. AB - Myofibrillar proteins in muscles of the claws and abdomen of lobster, Homarus americanus, and the claws of fiddler crab, Uca pugnax, and land crab, Gecarcinus lateralis, have been analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fibers contained numerous isoforms of structural and regulatory proteins in assemblages correlated with fiber type. One fast (F) and two slow (S1 and S2) fibers were identified. All F fibers possessed two isoforms of paramyosin (P1 and P2), while all slow fibers, with the exception of Uca major claw, contained only the P2 variant. S1 and S2 fibers were distinguished by the distribution of a large isoform of troponin-T (T1; Mr = 55,000); S2 fibers in all three species contained T1 in addition to one or two smaller-molecular-weight variants usually associated with S1 fibers. In order to determine whether the slow fibers differed in histochemical properties, land crab claw closer muscle was cryosectioned and stained for myofibrillar ATPase and NADH diaphorase activities. Most S2 fibers had lower ATPase and higher NADH diaphorase activities than S1 fibers, which indicated that S2 fibers had a lower rate of contraction and were more fatigue-resistant than S1 fibers. It is proposed that the S1 and S2 fibers defined by biochemical and histochemical criteria are identical to the slow-twitch and tonic fibers, respectively characterized physiologically. PMID- 2968439 TI - In vivo receptor binding, neurochemical and functional studies with the dopamine D-1 receptor antagonist SCH23390. AB - A series of in vivo experiments were undertaken, relating functional (motor activity, body temperature), dopamine (DA) receptor binding and neurochemical (catecholamine synthesis and utilization, DA release) aspects of the pharmacology of SCH23390 in the rat. The compound inhibited the locomotor hyperactivity, but not the hypothermia, induced by the potent DA stimulant DP-5,6-ADTN. Interstingly, SCH23390 simultaneously failed to displace DP-5,6-ADTN from its binding sites in the rat striatum--used as a direct in vivo biochemical index of DA (D-2) receptor interaction. The spontaneous locomotion in non-pretreated rats was likewise inhibited by SCH23390. The locomotor-suppressive action, but not the DP-5,6-ADTN-displacing capacity of the D-2 blocker haloperidol was significantly enhanced by SCH23390, suggesting that motility can be suppressed by either enhanced D-1 or D-2 (postsynaptic) receptor blockade, but also that the D-1 and D 2 sites involved may be physically distinct. SCH23390 only slightly altered in vivo neurochemical of DA synthesis, release and nerve-impulse flow, indicating that, while similar in suppressing dopaminergic behaviour, the D-1 antagonist is less effective than traditional neuroleptics as an activator of DA neuronal feedback mechanisms. The weak increases of DA synthesis and release nonetheless obtained were equal in magnitude (30-40%) in the limbic vs. striatal brain areas; also in this respect, SCH23390 thus differs from classical neuroleptics, which generally display more marked effects in the striatum than in limbic tissue. No major changes in the in vivo indices of NA synthesis and utilization (or in 5-HT synthesis) were found after SCH23390 administration, by and large supporting the DA receptor specificity of the compound. In summary, the studies demonstrated that SCH23390 can offset and accentuate, respectively, behavioural consequences of D-2 receptor stimulation and blockade. Importantly, at the same time no direct interaction at the level of D-2 DA receptor sites in the striatum was detected. Only slight, D-2 antagonist-like, changes in neurochemical indices of dopaminergic activity were observed after D-1 receptor blockade by means of SCH23390. With regard to DA agonist hypothermia, SCH23390 was without effect per se, but (at a high dose) attenuated the action of the D-2 antagonist haloperidol. The observations may indicate that the complex interactions between central D-1 and D-2 receptor-controlled mechanisms that influence behaviour, neurochemistry, and possibly autonomic nervous expression, are not identical. PMID- 2968440 TI - Immunohistochemical study of Alzheimer's disease using antibodies to synthetic amyloid and fibronectin. AB - Etiology and source of amyloid deposition in senile plaques of Alzheimer's disease (AD) are still unknown. In order to know whether or not fibronectin (Fn), an adhesive glycoprotein, is related to the amyloid deposition in the senile plaque, we conducted immunohistochemical studies using polyclonal anti-Fn and affinity-purified anti-amyloid component (Affi 28). Affi 28 was made by immunizing a rabbit against the synthetic peptide corresponding to residues 1-28 of the amyloid core protein reported by Masters et al. (1985). According to this study, four points became clear. First, Affi 28 is able to stain the subpial regions of AD as well as cerebrovascular amyloid and amyloid plaque cores. Second, it is suggested either that the etiology and source of neurofibrillary tangles and Pick body is distinct from that of the senile plaque or that any Affi 28 determinants of neurofibrillary tangles and Pick body are obscured sterically. Third, Affi 28 is useful to distinguish the senile plaque from the amyloid plaque of Creutzfeldt-Jakob disease. Last, there is no association between the amyloid in the senile plaque and Fn, at least immunohistochemically. The absence of Fn in the senile plaque suggests that Fn may not be requested for the deposition of amyloid fibrils. PMID- 2968441 TI - Ultrastructural observations on myelinated fibres in experimental diabetes: effect of the aldose reductase inhibitor ponalrestat given alone or in conjunction with insulin therapy. AB - Six groups of rats were studied over a 12-week period: onset and end controls, untreated diabetics, ponalrestat-treated diabetics, insulin-treated diabetics, and diabetics treated with ponalrestat and insulin. The concentrations of glucose, sorbitol and fructose significantly increased and that of myo-inositol significantly decreased in the sciatic nerve of untreated diabetic animals. Ponalrestat administration completely normalized sorbitol levels and partially corrected fructose and myo-inositol concentrations without altering nerve glucose levels. The biochemical abnormalities were also corrected in both the insulin treated and insulin and ponalrestat-treated diabetic animals. Myelinated fibre cross-sectional areas and axonal areas were significantly less in the tibial nerve of diabetic animals as compared with age-matched controls. Insulin treatment partially corrected the reduction in fibre and axonal area but teased fibre preparations showed an excess of axonal degeneration as compared with controls, untreated diabetics and ponalrestat-treated diabetics. Ponalrestat given alone or in conjunction with insulin therapy did not correct the reduction in fibre or axonal area and single isolated fibres from diabetic animals treated with ponalrestat and insulin showed a marked excess of axonal degeneration, probably related to hypoglycaemia. The study fails to reveal any significant beneficial effect of aldose reductase inhibition on the structural abnormalities in peripheral nerve in experimental diabetes. PMID- 2968443 TI - Origin of the atriopeptin-like immunoreactive innervation of the paraventricular nucleus of the hypothalamus. AB - The paraventricular nucleus of the hypothalamus (PVH) contains a prominent collection of varicose atriopeptin-like immunoreactive (APir) fibers. We have used immunohistochemistry and fluorescent retrograde tracers to investigate the origin of these fibers. All parts of the PVH contain APir fibers. The densest collections are found in the periventricular area and in the parvocellular components of the nucleus. Somewhat smaller numbers of fibers are found within the borders of the magnocellular part of the PVH. Following the injection of fluorescent retrograde tracers into the PVH and subsequent immunohistochemical staining, numerous retrogradely labeled APir neurons were observed in the anteroventral periventricular nucleus (AVPV), adjacent to the anteroventral tip of the third ventricle. Smaller groups of retrogradely labeled APir neurons were observed in the ventromedial part of the bed nucleus of the stria terminalis, the pedunculopontine and laterodorsal tegmental nuclei, and the medial part of the nucleus of the solitary tract. The APir projection from the AVPV to the PVH is of particular interest in view of the abundant evidence implicating both regions in the regulation of the fluid and electrolyte balance and blood pressure. AP may serve as a central neuromodulator as well as a circulating hormone in cardiovascular regulation. PMID- 2968442 TI - Three-dimensional sensitivity and caudal projection of neck spindle afferents. AB - 1. We recorded from neck muscle spindle afferents in the C2 dorsal root ganglion of the decerebrate cat using floating electrodes. The afferents presumably innervated mainly ventral and ventrolateral perivertebral muscles, and sternocleidomastoid. Stimuli consisted of combinations of rotatory head movements about the roll/pitch or pitch/yaw axes. An important difference from our earlier experiments (10) was the addition of yaw movement to the stimulus paradigm making possible a three-dimensional analysis of afferent behavior. 2. For each afferent we determined the most effective direction of tilt (orientation of the response vector) in three dimensions by using sinusoidal stimuli that combined pitch and roll, or pitch and yaw, or by measuring the gains to responses to roll, pitch, and yaw rotation. 3. Most afferents were sensitive to rotation around all three axes; pitch and yaw were usually more effective than roll. There was no indication of clustering of response vectors, as might be expected if the receptors were located in a small number of muscles each of which has receptors aligned in a homogeneous direction. 4. The responses of afferents were further studied using sinusoidal and trapezoidal stimuli aligned as closely as possible with the orientation of their response vector. The availability of the yaw stimulus made receptor classification based on response linearity, gain, and dynamic index more reliable than in our earlier experiments (10). 5. Muscle spindle responses were divided into three categories: A, B, and ambiguous. The evidence suggests that category A are probably spindle primary receptors and category B are secondaries. Ambiguous receptors have intermediate properties. 6. The caudal projection of spindle afferents was examined by delivering antidromic stimuli with a movable electrode on the surface of the ipsilateral dorsal column. Eighteen percent of the afferents projected to C4, and 14% as far as C5. Long caudal projections can be found in A, B, and ambiguous receptors with a range of directional sensitivities. 7. The evidence suggests that C2 spindle afferents make synapses in the midcervical segments with interneurons and propriospinal neurons that are part of the intraspinal pathway of the tonic neck reflex. PMID- 2968444 TI - Expression of transferrin receptors on lymphocytes: its correlation with T helper/T-suppressor cytotoxic ratio and rejection in heart transplant recipients. AB - Forty-one heart transplant recipients were monitored serially for the expression of transferrin receptors and T-helper/T-suppressor cytotoxic ratios on circulating lymphocytes during the hospitalization periods after heart transplantations (60.5 +/- 18.9 days). These values were retrospectively correlated with the patients' clinical status with respect to rejection and infection. During clinically stable periods the average values of percentage of transferrin receptor-positive lymphocytes and T-helper/T-suppressor cytotoxic ratios were 5.9 +/- 4.3 and 1.5 +/- 1.0, respectively. The percentage of transferrin receptor-positive lymphocytes increased to a level of 12.0 +/- 5.4 (p less than 0.001) during the early prerejection phase and remained at this level throughout the rejection period. T-helper/T-suppressor cytotoxic ratios increased to 1.96 +/- 0.92 during the early prerejection phase (p less than 0.05), peaked at 2.30 +/- 1.21 during the late prerejection phase (p less than 0.01), but began to decline by the rejection period. After rejection treatment percentage of transferrin receptor-positive lymphocytes decreased to 8.4 +/- 5.3 (p less than 0.05), and T-helper/T-suppressor cytotoxic ratios decreased to normal levels. In contrast, in patients with infectious complications, a remarkably elevated percentage of transferrin receptor-positive lymphocytes (20.7 +/- 11.7) and relatively low T-helper/T-suppressor cytotoxic ratios (1.3 +/- 0.5) were noted. The data show an association between the clinical status, such as rejection and infection, and these immunologic measurements as transferrin receptor-positive lymphocytes and T-helper/T-suppressor cytotoxic ratios in heart transplant recipients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968445 TI - OKT4 epitope deficiency as a cause of reduced "helper" T cells in children at risk for human immunodeficiency virus infection. PMID- 2968446 TI - Nonspecific and specific immune responses in a child with Down's syndrome and her sibling. A case report. AB - In a child with Down's syndrome (DS) and her sibling, host immune responses were evaluated under experimental gingivitis conditions. The children live in the same environment under identical conditions. In the DS child an earlier and more extensive gingival inflammation than in her sibling had been observed. Investigation of nonspecific host defense mechanisms revealed identical results in both children for the phagocytosis and intracellular killing of Candida albicans by polymorphonuclear leukocytes in crevicular washings (CR-PMNs), in blood (PB-PMNs) and blood monocytes. Furthermore, CR- and PB-PMNs were able to secrete identical amounts of hydrogen peroxide upon stimulation. The chemotactic response of PB-PMNs in the DS child was impaired, however. The results of the studies performed on parameters of specific host defense mechanisms showed low blastogenic responses to phytohemagglutinin (PHA) and pokeweed (PWM) by lymphocytes of the DS child as compared with her sibling. Also a lack of immune regulation leading to prolonged helper/inducer cell activation on a local (gingival) and circulation level and a less pronounced T-cell depression in PB were shown. Together, these differences observed in specific and nonspecific host response mechanisms may be responsible for the earlier and more extensive gingival inflammation found in the DS child. PMID- 2968447 TI - Biologic response to chronic blockade of vasopressin receptors in Sprague-Dawley rats. AB - The renal tubular arginine vasopressin receptor antagonist, d-(CH2)5-D Tyr(Et)VAVP, is a potent inhibitor of the vasopressin-induced stimulation of adenylate cyclase in rat renal medullary homogenates in vitro. In acute experiments in vivo, this antagonist increased urine volume and decreased urine osmolality after i.v. or s.c. administration in normally hydrated or dehydrated Sprague-Dawley rats. It did not show any effects in water-loaded rats. The duration of action of the antagonist was between 3 to 4 hr. Chronic i.v. infusion or repeated s.c. injections did not result in a persistent diabetes insipidus. A transient rise in water excretion was followed by a progressive normalization. The marked initial water loss was fully compensated for by an increased water intake so that plasma volume and extracellular fluid volume remained unchanged. After 1 week of treatment with the antagonist, glomerular filtration rate and plasma renin activity were not significantly different from base-line values. Only small functional deficits in renal concentrating capacity became manifest when drinking water was withheld. It is possible that the activation of endogenous compensatory mechanisms restored water balance during chronic arginine vasopressin receptor blockade. An intrinsic agonism of this antagonist, which was not detectable in acute experiments, might have contributed to the normalization of water balance by limiting the maximum anti-antidiuretic effects of renal tubular arginine vasopressin receptor blockade. PMID- 2968448 TI - Evaluation of the pharmacologic properties of a vasopressin antagonist in Brattleboro rats. AB - The arginine vasopressin (AVP) analog d-(CH2)5-D-Tyr(Et)VAVP is a potent competitive antagonist of AVP at renal tubular AVP receptors. In Sprague-Dawley rats, this compound induces diuresis after single injections but only a transient diabetes insipidus-like state during continuous infusion. To further evaluate the pharmacologic profile of d-(CH2)5-D-Tyr(Et)VAVP, the present experiments were performed in Brattleboro rats homozygous for hereditary hypothalamic diabetes insipidus. In these rats, acute and chronic administration of the antagonist induced significant antidiuretic effects. These agonistic effects persisted for up to 4 days after single injections and for more than 2 weeks after stopping continuous infusions. The antidiuretic effects of the antagonist during chronic administration were indistinguishable from those of AVP replacement. When the renal tubular AVP receptor antagonist was infused into diabetes insipidus rats that had received AVP for 1 week, it induced a transient rise in water intake. However, the peak values after administration of the antagonist were much lower than after AVP withdrawal. These observations suggest that d-(CH2)5-D-Tyr(Et)VAVP has substantial agonistic properties that are not detectable in Sprague-Dawley rats except for limiting the compound's maximum anti-antidiuretic efficacy. These agonistic effects together with endogenous compensatory mechanisms may allow Sprague-Dawley rats to maintain a normal water balance during the continuous administration of d-(CH2)5-D-Tyr(Et)VAVP. PMID- 2968449 TI - Characterization of [5,6-3H]SQ 29,548 as a high affinity radioligand, binding to thromboxane A2/prostaglandin H2-receptors in human platelets. AB - The binding of 5,6-3H(1S-[1 alpha, 2 beta(5Z), 3 beta, 4 alpha])-7-[3-([2 [(phenyl amino)carbonyl]hydrazino]methyl)-7-oxabicyclo[2.2.1]hept-2-yl]-5- heptenoic acid to receptors in human washed platelets (WP) and platelet membranes (PM) was characterized with regard to kinetics, saturability and competitive inhibition by putative thromboxane A2/prostaglandin H2 (TP)-receptor ligands. Specific binding of [3H]SQ 29,548 routinely amounted to 90 to 97% of total binding. The rate of association was 1.6 x 10(7) and 2.5 x 10(7) M-1 x min-1 in WP and PM, respectively. The corresponding rate of dissociation was 0.07 and 0.12 min-1, resulting in dissociation constants of 4.1 and 5.8 nM in WP and PM, respectively. Saturable binding to a single class of receptors indicated a receptor density of 2633 fmol/mg of protein in WP (1394 receptors/platelet; kd, 4.5 nM) and 1466 fmol/mg of protein in PM (kd, 11.3 nM). Specific binding of [3H]SQ 29,548 was inhibited by five antagonists (high/low affinity kd values in nanomolar), SQ 29,548 (WP, 5.2; PM, 7.3), SQ 28,668 (WP, 32; PM, 73), SQ 30,741 (WP, 28; PM, 50), BM 13,177 (WP, 140; PM, 4834) and BM 13,505 (WP, 5/379; PM, 11). Two agonists, U 44069 and U 46619, inhibited the binding in a biphasic manner, indicating binding to two receptor sites (approximately 20/80%) with kd values of 4/72 and 4/170 nM, respectively, in WP and 7/136 and 19/502 nM, respectively in PM. The demonstrated high affinity binding of [3H]SQ 29,548 to human platelet TP-receptors should make this radioligand a suitable and potentially useful tool in future studies of function, structure and regulation of TP-receptors. PMID- 2968450 TI - Tensile strengths of composite core materials containing added colorants. PMID- 2968451 TI - Post-transplant renal artery stenosis-percutaneous transluminal angioplasty (report of 3 cases). PMID- 2968452 TI - V1-vasopressin receptors in the septum of the rat brain. Electrophysiological evidence. AB - In slices from the rat brain, extracellular recordings were obtained from single neurones located in the lateral septum, an area known to receive a vasopressinergic innervation. Approximately half of the neurones tested responded to vasopressin by a concentration-dependent increase in firing rate, the lowest effective concentration being in the order of 2 nM. The effect of vasopressin was blocked by a synthetic structural analogue possessing vasopressor and oxytocic antagonistic properties on peripheral vasopressin and oxytocin receptors. Oxytocin had a weak effect in firing septal neurones, whereas a selective oxytocic agonist was totally ineffective. The action of vasopressin on neuronal firing was mimicked by a vasopressor agonist (Phe2-Orn8-VT) but not by a selective antidiuretic agonist (dDAVP). These results indicate that the vasopressin receptors present in rat septum are V1 (vasopressor type) rather than V2 (antidiuretic type) receptors. In addition, we conclude that these receptors, when occupied, lead to increased firing of lateral septal neurones. PMID- 2968453 TI - Renal V2 vasopressin receptor proteins: identification and enrichment. AB - The synthesis of the tritium labelled photoreactive analogue of 1-deamino vasopressin [1-(3-mercaptopropionic acid, 8-(N6-4-azido-phenylamidino)-lysine] vasopressin is described. This analogue retains a high affinity for hepatic V1 and renal V2 vasopressin receptors (apparent dissociation constant KD approximately 1-2 nM). A membrane protein from bovine kidney and pig kidney with an apparent relative molecular mass (Mr) of 30,000 was preferentially labelled and with lower yield a protein band with a Mr-value of 50,000 to 60,000. The photolabelled 30,000-Mr protein from bovine kidney was enriched by size-exclusion chromatography and by reversed-phase-high-performance liquid chromatography. PMID- 2968455 TI - Problems of compensation in occupational asthma and alveolitis. PMID- 2968454 TI - Laparoscopic sterilization. A comparison of current techniques. AB - Current techniques of laparoscopic sterilization--bipolar electrocoagulation, endocoagulation, silicone ring, Hulka clip and Filshie clip--were analyzed from a large pool of international data. Technical difficulties have been encountered much less with electrocoagulation than with the silicone ring, Hulka clip or Filshie clip. There have been fewer technical failures with bipolar electrocoagulation and endocoagulation than with the ring or clip. Surgical complications have been the fewest with bipolar electrocoagulation and endocoagulation but have been more serious (electrical burns). The Hulka and Filshie clips have had lower complication rates than has the silicone ring. The greater technical difficulties and failures and greater surgical complications with clips and rings have been due to technical problems in applying them in cases of tuboperitoneal pathology. The pregnancy rates from method failure have been acceptably low with all the methods. Ectopic pregnancies are a serious problem with bipolar electrocoagulation and endocoagulation. The pregnancy rates after reversal surgery have been highest with the Hulka and Filshie clip. Electrical methods seem to be preferable in women with pelvic adhesions and tubal pathology. Clips seem to be preferable in women who may be candidates for future sterilization reversal. PMID- 2968456 TI - Preparation of 7-oxaaporphine derivatives and evaluation of their dopaminergic activity. AB - A series of 7-oxaaporphine derivatives was prepared. The compounds were evaluated as dopaminergic agents. None of them showed either affinity for dopamine receptors or activity in vivo in the climbing behavior (mice) and turning behavior (6-hydroxydopamine-lesioned rats) tests. The lack of activity is tentatively related to the effect of the oxygen atom on the pKa of these molecules. PMID- 2968457 TI - The effect of campylobacter lipopolysaccharide on fetal development in the mouse. AB - Purified lipopolysaccharide (LPS) obtained from isolates of Campylobacter fetus ss. fetus and Campylobacter jejuni impaired fetal development when administered to mice on day 13 of pregnancy. Strikingly more fetal resorption was produced by C. jejuni LPS than by similar amounts of C. fetus ss. fetus LPS. Three of the four Campylobacter strains examined produced LPS that had no effect on maternal health, but LPS from one C. jejuni strain killed all of the mice to which it was administered. PMID- 2968458 TI - T-cell immunoregulation in patients with inflammatory bowel disease. II. Enhanced suppressor T-cell activity in ulcerative colitis. AB - In a recent study, we have shown that peripheral blood B cells from patients with ulcerative colitis (UC) synthesized less immunoglobulin (Ig) in co-culture with autologous T cells than normal adults' B cells. When UC patients' T cells were co cultured with normal adults' B cells, Ig synthesis was significantly decreased as compared with normal controls. In contrast, Crohn's disease (CD) patients' B and T cells functioned normally. In the present study, the activity of suppressor T cells in patients with UC and CD was determined. Peripheral blood B and T cells with monocytes were obtained from patients and normal adults of the same age and sex, and co-cultured for 10 days with pokeweed mitogen (PWM). Suppressor T-cell function was measured in mixed co-culture assays in which graded numbers of normal or patient's T cells were added to normal adults' B and T cells with PWM. Immunoglobulins (Ig) M, G and A were measured in culture supernatants using a sensitive enzyme-linked immunosorbent assay. The quantity of Ig present in the culture supernatants was determined from a standard curve. T cells from UC patients significantly decreased immunoglobulin production by control B and T cells (IgM and IgA, p = 0.02; IgG, p = 0.01). In contrast, addition of T cells from CD patients produced no significant differences. Complement mediated, monoclonal OKT8 antibody directed cell lysis revealed that the inhibition observed with UC patients' T cells in co-culture was due to a T8+ suppressor T cell. The degree of inhibition of immunoglobulin synthesis did not correlate with disease activity, duration of illness, location of disease, or corticosteroid treatment. Thus, patients with ulcerative colitis display enhanced suppressor T cell activity in peripheral blood while patients with CD show normal helper and suppressor T-cell functions. These results provide evidence supporting a role for altered immunoregulatory activity in the pathogenesis of ulcerative colitis. PMID- 2968460 TI - What we should expect from a lateral chest radiograph. PMID- 2968459 TI - T-cell immunoregulation in patients with inflammatory bowel disease. I. Differential helper T-cell function in ulcerative colitis and Crohn's disease. AB - The role of helper T-cells in patients with ulcerative colitis (UC) and Crohn's disease (CD) was investigated. Peripheral blood B- and T-cells with or without monocytes were obtained from patients and normal adults of the same age and sex, and co-cultured for 10 days with pokeweed mitogen (PWM). Immunoglobulins (Ig) M, G and A were measured in the culture supernatants using a sensitive enzyme-linked immunosorbent assay. The quantity of Ig present in the culture supernatants was determined from a standard curve. Immunoglobulins M, G and A synthesis and secretion by B-cells in the presence of T-cells required monocytes and PWM. The data indicate that co-cultures of heterologous normal adult T-cells and B-cells with PWM did not significantly affect Ig synthesis as compared with autologous cultures. Autologous cultures of CD patients' B- and T-cells were found not to be significantly different from normals in their capacity to synthesize Ig. In contrast, autologous UC patients' B- and T-cells were found to be significantly less effective as compared with normal adults' co-cultures in the synthesis Ig. When CD patients' T-cells were in co-culture with normal adults' B-cells, Ig synthesis was maintained. However, a marked diminution in Ig synthesis was seen when UC patients' T-cells were used in co-culture with normal adults' B-cells. The degree of inhibition of immunoglobulin synthesis did not correlate with disease activity, duration of illness, location of disease, or corticosteroid treatment. These results suggest that patients with ulcerative colitis have an altered helper T-cell population while CD patients' T-cells are either normal or hyperactive in the capacity to provide helper function in PWM-induced immunoglobulin secretion by peripheral blood B-cells. PMID- 2968461 TI - [A survey of clinical features of polychlorinated biphenyls poisoned mothers and their transplacental Yu-Cheng babies]. PMID- 2968462 TI - [Thermal effects of argon laser and hematoporphyrin derivatives on subcutaneous tumor tissue in mice]. PMID- 2968463 TI - Childhood disability: a study from a tribal block of South Rajasthan, India. PMID- 2968465 TI - Detection thresholds of nonocclusive intestinal hypoperfusion by Doppler ultrasound, photoplethysmography, and fluorescein. AB - Because clinical assessment of bowel viability is unreliable, other methods of determining intestinal perfusion have been recommended. Since none of these quantifies intestinal blood flow, we measured flow at the detection thresholds of Doppler ultrasound, photoplethysmography, and intravenously administered fluorescein, perfused the intestines at these threshold levels, and assessed histologic evidence of ischemic damage. The intestines of five anesthetized dogs were perfused for 4 hours via an in-line pulsatile extra-corporeal circuit assembled between the iliac and superior mesenteric arteries at either relatively physiologic (approximately 20 ml/min/kg body weight) levels or reduced levels representing the flow detection thresholds of Doppler ultrasound or photoplethysmographic probes (approximately 4 ml/min/kg). Intravenously administered fluorescein was detected at even lower perfusion levels (approximately 2.1 ml/min/kg). Clear-cut ischemic changes were documented histologically in all subjects perfused at Doppler/PPG flow detection thresholds but in none of those perfused at normal levels. We conclude that threshold blood flow detection by any one of these methods, especially fluorescein, occurs at levels inadequate to guarantee tissue viability. PMID- 2968464 TI - Experimental colonization of a polyester vascular graft with Staphylococcus aureus: a quantitative and morphologic study. AB - Colonization of a polyester (Dacron) vascular graft by Staphylococcus aureus 209P R was studied. Twenty-five dogs had thoracoabdominal aortic bypass. After intervals of 2 hours (three dogs), 8 days (five dogs), 1 month (six dogs), 2 months (six dogs), or 6 months (five dogs), a bacteremic challenge was produced by intravenous injection of 6 x 10(8) colony-forming units of S. aureus. Two hours later grafts were removed and cut into 10 fragments, each submitted to bacterial counts and scanning electron microscopic studies. Results of bacterial counts were expressed in colony-forming units (CFU) per square centimeter of graft segment (median [lower to upper quartiles]). Normal canine aortas (n = 2) used as controls trapped no bacteria. Colonization of Dacron grafts varied according to the duration of graft function (p less than 0.01): after 2 hours, 4416.5 CFU (1158 to 9073 CFU); after 8 days, 1515 CFU (963 to 2893 CFU); after 1 month, 199 CFU (86 to 538 CFU); after 2 months, 615 CFU (243 to 1407 CFU); and after 6 months, 1 CFU (1 to 5 CFU). Heavily colonized fragments were observed for duration of graft function of 2 months or less, whereas at 6 months all the fragments trapped fewer than 50 CFU/cm2 of graft segment. Scanning electron microscopy showed that colonization was closely associated with healing. Staphylococcal entrapment was related to the amount of fibrin deposits, which were especially abundant where the thrombotic matrix was unorganized and on bare polyester filaments. Graft colonization is especially to be feared in the first weeks after graft implantation, an observation which may help to define guidelines for preventing hematogenous vascular graft infection. PMID- 2968466 TI - An experimental study of a new sutureless intraluminal graft with an elastic ring that can attach itself to the vessel wall. A preliminary report. AB - A new sutureless intraluminal graft was developed with an elastic ring made of a flat spring. The diameter of the ring could be reversibly reduced by compression. The sutureless intraluminal graft with an elastic ring can attach itself to the vessel wall by elastic expansion of the ring. The elastic-ring graft was implanted in the descending thoracic aortas of nine dogs and was evaluated histologically and angiographically at different intervals from 18 to 150 days. No complication such as detachment of the ring, aortic rupture, stenosis, or aneurysmal dilatation was observed. With the new graft neither ligation nor posterior aortic wall dissection is necessary, and no anastomotic stenosis occurs. This graft is applicable even if the diameter of the aorta is small. Therefore the elastic-ring sutureless intraluminal graft promises theoretic advantages over sutureless methods that use tape ligation. PMID- 2968467 TI - Percutaneous transluminal angioplasty. PMID- 2968469 TI - The appropriateness of performing coronary artery bypass surgery. AB - Information about how appropriately procedures are performed is vital to the understanding of the impact of technology and to the success of efforts to channel its use appropriately. While the efficacy of coronary artery bypass surgery has been addressed in several large-scale, randomized trials, there is little information about how appropriately the procedure is actually being used in the community. We determined the appropriateness of coronary artery bypass surgeries performed in three randomly chosen hospitals in a western state. We determined appropriateness by comparing data obtained from a detailed medical record review with a list of 488 indications. This list, developed by a national panel of physicians, covered all possible reasons for performing the procedure. Three hundred eighty-six cases from the years 1979, 1980, and 1982 were examined. Fifty-six percent of the surgeries were performed for appropriate reasons, 30% for equivocal reasons, and 14% for inappropriate reasons. The percentage of appropriate surgeries varied by hospital, from 37% to 78%, but did not vary by patient age. Eliminating the performance of inappropriate procedures may lead to reductions in health care expenditures or to improved patient outcomes. PMID- 2968470 TI - Postexposure prophylaxis of hepatitis B. PMID- 2968468 TI - Leads from the MMWR. Prevention of perinatal transmission of hepatitis B virus: prenatal screening of all pregnant women for hepatitis B surface antigen. PMID- 2968472 TI - [Decreased endothelial antithrombogenic activity in disseminated intravascular coagulation]. PMID- 2968471 TI - Differences in regulatory mechanisms of atrial and ventricular muscle contraction in bovine heart. AB - The purpose of this study was to characterize the regulatory mechanisms of atrial muscle contraction. Natural actomyosin (NAM) and tropomyosin-troponin (TM-TN) complex were prepared from atrial and ventricular muscle of the same bovine heart. The results were as follows: (1) Atrial NAM was more sensitive to Ca2+ than was ventricular NAM: the pCa required for 50% ATPase activation was 5.96 +/- 0.10 vs. 5.63 +/- 0.07, (mean +/- SE; n = 6; p less than 0.01); (2) reconstitution of desensitized actomyosin of rabbit skeletal muscle plus atrial or ventricular TM-TN complex produced higher Ca2+ sensitivity in atrial muscle than in ventricular muscle: the pCa required for 50% ATPase activation was 6.48 +/- 0.10 vs. 6.23 +/- 0.15 (n = 3; p less than 0.05); (3) the amount of inorganic phosphate covalently bound to atrial NAM was equivalent to that bound to ventricular NAM; (4) SDS-polyacrylamide gel electrophoresis of the two NAMs revealed several protein bands of different mobility from 16,000 to 30,000 daltons; and (5) the superprecipitation response of atrial NAM was characterized by a stepwise change in turbidity after the addition of MgATP, in contrast to the biphasic pattern of ventricular NAM. These data suggest that the free Ca ion concentration required for atrial muscle contraction is lower than that required for ventricular muscle contraction and that the difference is attributable to differences in atrial and ventricular regulatory proteins. PMID- 2968473 TI - [Acute megakaryoblastic leukemia in early childhood; report of two cases]. PMID- 2968474 TI - [T8 lymphocytosis with rearrangement of the gene for the beta chain of the T cell receptor is found in a case of AMMoL in remission]. PMID- 2968476 TI - [Fatigue and low-back pain of freight-container tractor drivers. Correlation with ergonomic factors]. AB - An ergonomic checklist was prepared to examine safety and comfort of freight container tractors. With this checklist, 533 tractor drivers were asked to evaluate the tractors to which they were exclusively assigned. In order to check the objectivity and the validity of the results, the correlation between the evaluation of the items on the checklist and the corresponding dimensions on 10 tractors was studied. Also, a multivariate analysis based on quantification theory was done to examine the correlation between the ergonomic items on the checklist and the drivers' low-back pain or fatigue symptoms of the upper and lower extremities. The main results were as follows: 1) A high correlation was observed between the evaluation of items on the checklist and the corresponding dimensions of the tractors. 2) Many items on the checklist received a poor evaluations, indicating that there are many ergonomic problems with freight container tractors. 3) The analysis by the quantification theory showed high correlation ratios between the items in the checklist and the drivers' fatigue symptoms of the upper and lower extremities. These correlation ratios were: arm fatigue when holding the steering wheel (0.51), arm fatigue in operating the gearshift lever (0.65), arm fatigue from opening and closing the windows (0.46), leg fatigue from operating the pedals (0.58), and low-back pain during the last two months (0.31). These showed that more than 50% of the fatigue symptoms could be explained by the ergonomic aspects of the tractors. 4) The rate of dissatisfaction with the current tractors was 82.9%. According to the analysis by the quantification theory, the correlation ratio between the items in the checklist and the rate of dissatisfaction for the current tractors was 0.66. 5) The foregoing results indicate that ergonomic improvement of the freight container tractors is a matter of urgency. This should reduce the drivers' fatigue and elevate the rate of satisfaction with the tractors. 6) However, for the prevention of the drivers' low-back pain and fatigue symptoms, improvement of the ergonomic conditions of the tractors and the drivers' working conditions must be undertaken at the same time. PMID- 2968475 TI - [Evaluation of common carotid blood flow volume in elders studied with a two dimensional echographically guided ultrasonic blood flow-meter]. PMID- 2968477 TI - [Significance of pulmonary arterial pressure on the release of atrial natriuretic factor (ANF)]. PMID- 2968478 TI - Partitional measurements of circulation can be made between capillaries and arteriovenous anastomoses in the human finger. AB - We compared the responses of finger blood flow to local and/or whole body warming, as measured by laser Doppler flowmetry (PERIFLUX PF-1d) (LDF), and by venous occlusion plethysmography (BF). The subject's hands were immersed in a bath of water whose temperature (Tw) was initially set at 25 degrees C. The tested hand was kept at about 10 cm above heart level to facilitate the drainage of finger veins during measurement. The ambient temperature (Ta) was initially 20 degrees C. Then, measurements were made under the following conditions on separate days: 1) local hand warming only (LHW), Tw raised from 25 to 40 degrees C; 2) whole body warming only (WBW), Ta was raised from 20 to 35 degrees C; and 3) LHW and WBW combined (CW), LHW (Tw was raised to 35 degrees C and maintained) was followed by WBW. Results showed that both LHW and WBW increased LDF and BF. LDF signals correlated significantly with BF values in each condition (r = 0.932 0.955). However, the slope of the line showing the relationship of LDF and BF, determined by least squares linear regression analysis, was steeper in LHW than in WBW. In CW, the slope showed a bend, creating two different sections in the relationship. Below the bend the relation was steeper and showed a similar slope as that in LHW; above the bend, the relation was more gradual and a similar slope as that in WBW. No such bend was observed in measurements of the forearm skin in CW.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968479 TI - Myocardial interstitial fibrosis in experimental uremia--implications for cardiac compliance. AB - Experimental uremia is known to cause cardiac hypertrophy. In the present study we examined the effect of uremia with or without concomitant treatment of hypertension by the converting enzyme Ramipril (125 micrograms/day) on micromorphometric indices of cardiac interstitium at the light microscopical and ultrastructural level. In male SD rats, 21 days of uremia caused an increase of total heart weight (1040 +/- 73 mg wet wt vs. 871 +/- 81 in controls, P less than 0.05) with an increase of both right and left ventricular weight. This was accompanied by reduction of capillary cross-sectional area despite unchanged capillary length. The volume density (cm3/cm3) of cardiomyocytes was unchanged (0.881 +/- 0.01 vs. 0.871 +/- 0.016 in controls), but volume density of interstitial tissue (excluding capillary lumen) was significantly increased (0.042 +/- 0.011 cm3 interstitial tissue/cm3 total heart tissue vs. 0.019 +/- 0.007 in controls). This was associated with signs of activation of interstitial cells, that is, increased volume of interstitial cell nuclei and interstitial cell cytoplasm. Concomitantly, a significant increase of volume density of non cellular interstitial ground substance was found which was not normalized by antihypertensive treatment using Ramipril. After three months of uremia, electron microscopy showed collagen fiber deposition in the interstitium. Comparable interstitial fibrosis was not observed in hearts of rats with renovascular (one clip-two kidney) hypertension. It is concluded that uremia increases myocardial interstitial ground substance by mechanisms independent of hypertension. The data may be relevant for recent findings of diastolic heart malfunction secondary to impaired compliance in uremic patients. PMID- 2968480 TI - [Prevention of complications involving the abdominal wall in patients with acute appendicitis]. PMID- 2968481 TI - The effects of corticotropin, opioid peptides and crude pituitary extract on the production of dehydroepiandrosterone and corticosterone by mature rat adrenal cells in tissue culture. AB - In order to study the steroidogenic response to pituitary factors, a technique of monolayer tissue culture of mature female rat adrenal cells was used. During the first 24 h, rat adrenal cells produced dehydroepiandrosterone (DHEA) and small amount of corticosterone but in the absence of corticotropin (ACTH), the release of these two steroids were reduced to very low levels. The addition of synthetic alpha-ACTH-(1-24) [0.01-100 ng/ml] elicited a marked increase in the production of both steroids. This stimulating effect was not observed when synthetic methionine and leucine-enkephalins (1-100 ng/ml), human beta-endorphin (1-100 ng/ml) or human beta-lipotropin (1 ng/ml), were added to the culture medium. When these peptides were added concomitantly with alpha-ACTH (1-24) at half of the maximum response dose (1 ng/ml), no synergistic effect upon DHEA and corticosterone production was shown. The addition of crude extract from rat pituitary gland (1-100 ng/ml) with or without alpha-ACTH-(1-24) definitely showed both a stimulatory and synergistic effect upon the production of these two steroids. Furthermore, the ratio between DHEA production and corticosterone production was significantly higher when crude extract of the pituitary gland was given alone or concomitantly with alpha-ACTH(1-24) than when alpha-ACTH(1-24) was given alone. These data suggest the existence of a still undefined pituitary adrenal androgen stimulating which may preferentially stimulate DHEA production over corticosterone production. PMID- 2968482 TI - Dehydroepiandrosterone therapeutics: acetylation of DHA in mouse liver. AB - This investigation was designed to evaluate the possibility that the therapeutic benefits of dehydroepiandrosterone (DHA)-feeding in mice is mediated by (1) a metabolite of DHA formed in liver or (2) by way of the obligatory hepatic metabolism of DHA fed in large amounts. We found that the pattern of metabolism of DHA is strikingly different when DHA in low (tracer) quantities is incubated with mouse liver compared with that found when DHA in high concentrations is incubated with this tissue. In the former case, the principal metabolites are sulfoconjugates and other polar compounds, e.g. hydroxylated products. In the case of DHA metabolism when the substrate is present in high concentrations (1 100 microM), the principal metabolite formed is delta 5-androstenediol. And in this case, there is the formation of very nonpolar metabolites, which we have identified as the acetates of DHA and delta 5-androstenediol. We find further that the acetates are formed by a transacetylation mechanism in which performed acetylated compounds, e.g. pregnenolone acetate and ethyl acetate, can serve directly as co-substrates; but (at least in short-term incubations), Na+-acetate and acetyl CoA do not serve as co-substrates. We suggest that the therapeutic benefits of DHA-feeding in mice may be, in part, the result of alterations in hepatic intermediary metabolism that is obliged by the metabolism of DHA when this otherwise inert agent is fed in large amounts. Thus, DHA-feeding may serve to cause changes similar to those that are beneficial or therapeutic with dietary manipulations including caloric restriction. PMID- 2968484 TI - Aldosterone nuclear receptors in kidneys of chick embryo. AB - We have studied the properties of the nuclear receptors for aldosterone in kidneys of chick embryo. Aliquots of 0.4 M KCl nuclear extracts were incubated with [3H]aldosterone with or without 1 microM RU28362, a potent glucocorticoid analog. Scatchard analyses of binding data revealed two classes of binding sites with Ka of 0.26 and 0.03 X 10(9) M-1 and Nmax of 330 fmol and 620 fmol/mg DNA respectively. In presence of RU28362, however, we observed only a single class of binding sites with a Ka of 1.02 X 10(8) M-1 and a Nmax of 90 fmol/mg DNA. Competition studies performed in presence of RU28362 showed that aldosterone was the more effective competitor followed by corticosterone, progesterone, deoxycorticosterone, dexamethasone, cortisol, triamcinolone acetonide and cortisone. The nuclear complexes had a sedimentation coefficient in the area of 8 S which changed to 4-5 S in the presence of 0.4 M KCl. This effect of KCl was prevented by the addition of 10 mM sodium molybdate. Always in the presence of the glucocorticoid analog, by DEAE-c chromatography we observed a major specific aldosterone-binding fraction which was eluted with 0.2 M KCl. This fraction sedimented at 8.4 S in the absence of sodium molybdate and KCl. In the absence of RU28362, DNA-c columns retained only a small portion of the nuclear complexes which were eluted with KCl. These complexes sedimented, on sucrose gradient, at 4.6 and 3.1 S, whereas those which did not bind to DNA-c had a sedimentation coefficient of 8 S. In the presence of RU28362, the majority of bound [3H]aldosterone remained in the column flow-through fraction; when this fraction was further analyzed on DEAE-c, complexes were eluted with 0.2 and 0.3 M KCl. These data indicate that nuclear receptors for aldosterone are present in small number in kidneys of chick embryo and that they are mostly in the 8 S form. PMID- 2968483 TI - Interactions between 17 beta-estradiol and the hypothalamo-pituitary beta endorphin system in the regulation of the cyclic LH secretion. AB - This paper further substantiates the physiological role of beta-endorphin (beta END) in the control of the cyclic LH secretion and provides new data on the interactions between 17 beta-estradiol (17 beta-E2) and beta-END at both the hypothalamic and pituitary levels. At the hypothalamic level, during the estrous cycle in rats, beta-END concentrations were highest on diestrus I in the arcuate nucleus, median preoptic area and median eminence and lowest at the time of the preovulatory 17 beta-E2 surge on proestrus, before the subsequent preovulatory hypothalamic GnRH and plasma LH surges. Data obtained in ovariectomized 17 beta E2-treated ewes support the direct involvement of 17 beta-E2 in changes in beta END and GnRH concentrations in these hypothalamic areas. At the anterior pituitary level, in vitro results obtained using anterior pituitaries from the proestrus morning cycling female rat have shown that 17 beta-E2 strongly suppresses beta-END secretion and that GnRH stimulates the release of beta-END. Furthermore, marked fluctuations were observed for plasma beta-END throughout the menstrual cycle in the woman. Low beta-END concentrations were observed in the period preceding the LH preovulatory surge. Taken together, these results show that: (1) decreases in hypothalamic beta-END concentrations, which are controlled at least by circulating levels of 17 beta-E2, modulate GnRH synthesis and/or release and contribute to the mechanisms which initiate the LH surge; (2) anterior pituitary beta-END might be involved in the mechanisms which terminate the LH surge. PMID- 2968485 TI - Effects of androgen and antiandrogen on growth, morphology and synthesis of specific proteins in mouse mammary carcinoma cells. AB - Primary cell cultures derived from an androgen-dependent mouse mammary carcinoma, the Shionogi SC-115 tumor, display characteristic changes in growth, morphology and protein synthesis according to the presence or absence of testosterone. In the presence of testosterone, cell proliferation was increased and cells formed characteristic clones having no contact inhibition. Ultrastructural studies of cells showed close contacts of plasma membranes having little or no gap between cells. Some cells were related by bridges of extracellular matrix. Testosterone induced synthesis of several intracellular and secreted proteins was observed after [35S]methionine-labeling of cells, SDS-PAGE and autoradiography, as well as the disappearance of a protein in androgen-treated cells. In the absence of testosterone, cells grow as a monolayer, have contact inhibition and flattened morphology. The ultrastructurally observed cell-to-cell contacts were usually less intimate, showing spaces of irregular width between cells. None of the testosterone-induced proteins were observed in the absence of hormone. The antiandrogen cyproterone acetate, which by itself was inactive, completely suppressed the androgen-induced effects on growth, morphology and specific protein synthesis. Glycosylation of membrane proteins, as measured after labeling of cells with [3H]N-acetyl-D-glucosamine, was increased by approximately 30% in the presence of testosterone. A similar observation was made in situ by autoradiography on intact cells. Finally, we found that culture medium conditioned by testosterone-treated Shionogi cells had significant mitogenic activity on L-929 mouse fibroblasts. PMID- 2968486 TI - The effects of androgens and cortisol on the in vivo metabolism of oestradiol. AB - We have examined the effect of co-administration of dehydroepiandrosterone sulphate, 5-androstenediol or cortisol on the metabolic clearance rate of oestradiol (MCR-E2) and conversion of oestradiol to oestrone (CRE2E1). Previous studies have shown that these androgens influence the metabolism of oestradiol in vitro while cortisol alters the distribution of oestradiol in plasma. The MCR-E2 and CRE2E1 were measured after 2.5 and 5 h of [3H]oestradiol infusion with co infusion of androgen or cortisol starting after 2.5 h of tracer infusion. For one subject who did not receive co-infusion of another steroid no significant change in MCR-E2 or CRE2E1 occurred over the 5-h period. For other subjects, however, the MCR-E2 decreased by 18 +/- 7% (mean +/- SD) while the CRE2E1 increased by 45 +/- 12%. It is possible that these results are due to: changes in the distribution of oestradiol in plasma; differences in the metabolism of oestradiol bound to albumin or SHBG, or an effect of androgens or cortisol on the uptake of [3H]oestradiol by the liver. PMID- 2968487 TI - Glucocorticoid effects on the production and actions of immune cytokines. AB - The immunosuppressive actions of glucocorticoids form the basis for their use in treatment of autoimmune diseases and prevention of allograft rejection. However, the mechanisms responsible for glucocorticoid-induced immunosuppression are still poorly understood. It is now clear that glucocorticoids do not inhibit all aspects of the immune response and, in some cases, may enhance certain functions of immune effector cells. One example is that of the dramatic increase induced by IFN-gamma in the number of IgG Fc receptors on human mononuclear phagocytes, which is enhanced rather than inhibited by glucocorticoids. An aspect of the immune response which appears to be consistently suppressed by glucocorticoids is the production of immune cytokines. Since these hormones appear to be essential mediators for a vigorous immune response, inhibition of their production may be an effective way for glucocorticoids to block the immune response. PMID- 2968489 TI - The effect of atrial natriuretic peptide on acid-base balance in rats with chronic renal failure. AB - We explored the effects of 12-hour infusion of atrial natriuretic peptide (alpha rANP:rat, 1-28) on arterial acid-base balance, using 5/6 nephrectomized rats with chronic renal failure. Before the infusion, nephrectomized rats had a higher mean arterial blood pressure, greater urine volume, and lower creatinine clearance than the normal controls, but they did not show a significant difference in arterial hydrogen ion concentration (pH), plasma bicarbonate concentration (HCO3 ), partial pressure of carbon dioxide (PCO2), plasma base excess (BE), or plasma ANP concentration. alpha-rANP infusion produced a continuous blood pressure reduction in both nephrectomized and control rats. Urine volume and urinary sodium and potassium excretion tended to increase at 2-hour infusion, but not at 12-hour infusion. In the controls alpha-rANP significantly increased pH from 7.47 to 7.50, and decreased PCO2 by 14%. In contrast, in nephrectomized rats alpha rANP significantly decreased pH from 7.48 to 7.44, HCO3- by 13%, and BE from 0.07 to -3.22 meq/l. Rats with chronic renal failure had greater reduction in HCO3- than the controls (p less than 0.05). There was no difference in plasma ANP level between the two groups. Thus, it is indicated that the long-term infusion of alpha-rANP reduces pH in rats with chronic renal failure, thereby adversely affecting the acid-base balance. PMID- 2968488 TI - Pure uptake blockers of dopamine can reduce prolactin secretion: studies with diclofensine. AB - The effects of diclofensine, a pure dopamine (DA) uptake inhibitor on 1) 3H-DA uptake in rat arcuate-periventricular nucleus-median eminence synaptosomes, 2) basal and K+-evoked endogenous DA release from tuberoinfundibular dopaminergic (TIDA) neurons and 3) in vivo prolactin (PRL) secretion were studied. Diclofensine, in concentrations of 0.01, 0.1 and 1 microM caused a marked decrease of 3H-DA uptake. In addition, it was unable to stimulate basal endogenous DA release which, on the contrary, was elicited by d-amphetamine in the same concentration (50 microM). On the other hand, diclofensine (50 microM) caused a 3 fold enhancement of K+-evoked DA release. Finally, the compound, when administered in vivo to male rats, significantly reduced basal serum PRL levels. The results of the present study seem to indicate that the pharmacological blockade of DA uptake in TIDA neurons is a condition sufficient to cause a reduction of PRL release. PMID- 2968490 TI - Anti-curare effect of plasma from patients with thermal injury. AB - Following severe thermal injury, patients are resistant to non-depolarizing muscle relaxants. Although this resistance has been well documented clinically, little is known about its etiology. We have tested the hypothesis that circulating factors contribute to the decreased potency of neuromuscular blockers following burns. The potencies of d-tubocurarine (2 microM) or pancuronium (2 microM) dissolved in plasma from either burned or control human subjects were tested on the indirectly stimulated (0.2 Hz) rat phrenic nerve-hemidiaphragm preparation. The muscle relaxants produced less neuromuscular blockade when dissolved in plasma from burned patients than when they were dissolved in plasma from controls. Thus, circulating factors are involved in the decreased potency of non-depolarizing neuromuscular blocking drugs. PMID- 2968491 TI - [Work experience of a municipal council of paramedical workers]. PMID- 2968492 TI - [Automated prophylactic examinations in childhood]. PMID- 2968494 TI - Detection of motion using B1 gradients. AB - An NMR method which makes use of a radiofrequency (RF) field gradient to detect diffusion, perfusion, or flow is demonstrated. The technique is analogous to the detection of motion using pulsed B0 gradients. The simplest form of the experiment is as follows: A spatially imhomogeneous RF field generated by a surface coil produces dispersal of magnetization in the Y Z plane. After a delay, the RF field is applied within reversed phase to restore polarization along +Z, which is then sampled using an observe pulse or pulse sequence. If molecular motion occurs during the delay period the amplitude of Z magnetization will be reduced. The lengths of the RF pulses and the delay period are varied so that the effects of relaxation, flow, and diffusion or perfusion can be distinguished. The present study demonstrates the use of this method to detect slow fluid flow. Advantages of this method include the availability of large RF gradients, and the avoidance of eddy currents. It is suggested that the method may have application to the study of perfusion and flow in vivo. PMID- 2968493 TI - [The team form of organization and pay of the paramedical and junior medical personnel at the Gorki Province N. A. Semashko Clinical Hospital]. PMID- 2968495 TI - Increased synthesis of glycosaminoglycans of the cornea during fasting and refeeding in rats. AB - The effect of a 48 hour fast on sulfate incorporation of corneal mucopolysaccharides was investigated in the rat. After fasting for 24 hours, experimental and control rats were injected intraperitoneally with 35S-sulfate. After fasting an additional 24 hours, the rats were sacrificed and incorporation of the label into the acid mucopolysaccharide fraction of the cornea was determined. Incorporation of the label into the acid mucopolysaccharide fraction was significantly increased in the fasted rat cornea. Increased synthesis of corneal sulfated mucopolysaccharides during fasting in the rat contrasts with the well known decreased synthesis in cartilage under the same conditions. These findings suggest that corneal acid mucopolysaccharide synthesis is selectively preserved and/or increased during fasting. PMID- 2968496 TI - Increased leucocyte Na.K ATPase in obesity: reversal following weight loss. PMID- 2968497 TI - Changing patterns of groups at high risk for hepatitis B in the United States. PMID- 2968498 TI - Delineation of DNA sequences that are important for in vitro transcription from the adenovirus EIIa late promoter. AB - Late in infection, transcription of the EIIa gene is initiated primarily at map unit 72 of the adenovirus genome. A cell-free nuclear extract system was used to determine sequence elements important for the function of this late promoter. In such a system, the transcriptional activity of a circular template was found to be much higher (5- to 10-fold) than that of a linear template. The effect of template topology appeared to be dependent on two distal upstream elements with 5' boundaries located near -265 to -223 and -147 to -133 (in relation to the initiation site), since deletions of these regions reduced transcription of the circular template, in a stepwise fashion, to a level similar to that observed with the linear template. Further deletions revealed an element in the -116 region that appeared to be more important for transcription of the circular template (10-fold reduction) than for transcription of the linear template (3 fold reduction). Lastly, deletion of the TACAAA sequence in the -29 region resulted in further reduction in transcription, indicating that this element functions as a promoter in vitro. PMID- 2968501 TI - Cloning of methylated transforming DNA from Neurospora crassa in Escherichia coli. AB - An arg-2 mutant of Neurospora crassa was transformed to prototrophy with a pBR322 N. crassa genomic DNA library. Repeated attempts to recover the integrated transforming DNA or segments thereof by digestion, ligation, and transformation of Escherichia coli, with selection for the plasmid marker ampicillin resistance, were unsuccessful. Analyses of a N. crassa transformant demonstrated that the introduced DNA was heavily methylated at cytosine residues. This methylation was shown to be responsible for our inability to recover transformants in standard strains of E. coli; transformants were readily obtained in a strain which is deficient in the two methylcytosine restriction systems. Restriction of methylated DNA in E. coli may explain the general failure to recover vector or transforming sequences from N. crassa transformants. PMID- 2968500 TI - Adenovirus E1a ras cooperation activity is separate from its positive and negative transcription regulatory functions. AB - The E1a gene of adenovirus encodes two proteins, 289 and 243 amino acids long, which have positive (transactivator) and negative (enhancer repressor) RNA polymerase II transcriptional regulatory properties and cell transformation activities including cooperation with an activated ras gene. The E1a transforming functions more closely correlate with the repressor property than with transactivation in that both E1a proteins express the repressor and transformation functions while only the 289-amino-acid protein is an efficient transactivator. To understand whether the transcriptional regulatory activities of E1a are related to its ras cooperation activity, we generated a series of mutant E1a expression vectors by linker insertion mutagenesis of the 289-amino acid protein. Here we describe a new class of mutants which although defective for enhancer repression still can cooperate with the ras oncogene in cell transformation. The mutants are also defective in transcription transactivation. Our data suggest that enhancer repression and transformation via ras cooperation are separate E1a functions and that cooperation with ras does not rely on either of the RNA polymerase II transcription regulatory functions of E1a. We also show that mutations which inactivate enhancer repression are not confirmed to a single critical domain necessary for repression. We therefore propose that the integrity of the overall configuration of the E1a proteins is important for the repression activity. PMID- 2968499 TI - Unusual branch point selection in processing of human growth hormone pre-mRNA. AB - Intron A of the human growth hormone gene does not contain an A residue within 56 nucleotides preceding the 3' splice site. The analysis of the excised intron lariat revealed a C residue 28 nucleotides upstream from the 3' splice site as the major branch acceptor nucleotide. Two additional minor branched nucleotides were identified as U residues at positions -22 and -36. An adenosine substitution at position -22 results in lariat formation solely to this nucleotide. Therefore, C and U residues can function efficiently as natural branch acceptors, but an A residue is preferred if available in the proper region. In addition, the data strongly reinforce the importance of the distance constraint for lariat formation. To explain selection of the branch acceptor nucleotide, potential base pairing interactions of branch point sequences with the U2 RNA are discussed. PMID- 2968503 TI - The biological significance of specific antibody IgG subclass profiles. PMID- 2968504 TI - [Reactive cells in non-Hodgkin's malignant lymphoma]. PMID- 2968502 TI - Distribution of IgG subclasses among human autoantibodies to Sm, RNP, dsDNA, SS-B and IgG rheumatoid factor. AB - The IgG subclass distribution of human autoantibodies to Sm, double-stranded DNA (ds-DNA), ribonucleoprotein (RNP), SS-B (La), and IgG rheumatoid factor (RF) have been determined using sensitive ELISA or by indirect immunofluorescence on Crithidia lucilia in sera from patients with systemic lupus erythematosus (SLE), Sjogren's syndrome, and rheumatoid arthritis. For anti-Sm and anti-RNP, IgG1 was the predominant isotype. For anti-ds-DNA and anti-SS-B, IgG1 and a lesser contribution of IgG3 was found. In contrast, IgG1 and IgG4 were the predominant isotypes of human IgG RF. The preponderance of isotypes noted for these autoantibodies did not extend to the IgG subclass distribution for antibodies to trinitrophenol-bovine serum albumin (TNP), tetanus toxoid (Tet. tox.), pneumococcal polysaccharides (Pneumo), and group A streptococcal cell walls (Strep.). The restriction of human humoral responses as well as autoantibodies has both pathogenetic and immunoregulatory implications, and suggests that for these autoantibodies, T-cell-dependent responses, probably driven by antigen, are of importance. PMID- 2968505 TI - Contributions to the architecture and significance of the hepatic and perihepatic fibrous structures. PMID- 2968506 TI - Causal factors of the space device variability of the hepatic veins (v. v. hepaticae). PMID- 2968507 TI - Anastomoses in the liver drainage territories. PMID- 2968508 TI - Lymph node infarction and malignant lymphoma. PMID- 2968509 TI - Poorly differentiated ("insular") thyroid carcinoma of the ovary. PMID- 2968510 TI - On the Romanian priority of E. C. Craciun and Aug. Muresanu in the practice of renal needle puncture. PMID- 2968511 TI - Proceedings of the Eighth Romanian Congress of Normal and Pathological Morphology and of the Jubilee Scientific Session of the Victor Babes Institute's centenary. Bucharest, September 22-24, 1987. PMID- 2968512 TI - Elimination of graft hemorrhage in heparinized patients with a collagen-coated Dacron prosthesis. PMID- 2968513 TI - Intragenic suppression in the uvrD gene of Escherichia coli. I. Temperature sensitive uvrD mutations. AB - A temperature-sensitive uvrD mutant, HD323 uvrD4, was isolated from the uvrD mutant HD4 uvrD3. The temperature sensitivity of the uvrD4 gene product was reversible. The suppressor mutation uvrD44 which rendered the uvrD3 mutant temperature-sensitive could be separated from the uvrD3 mutation by replacing the PstI fragment, which encodes the C-terminal half of the UvrD protein. The uvrD44 mutation was found to make host bacteria lethal at non-permissive temperatures only when cloned on a low copy vector pMF3. The nucleotide sequence of the uvrD3 and uvrD4 mutant genes was determined. The nucleotide change found in the uvrD3 at +1235, GAA to AAA, only alters the amino acid sequence from Glu at 387 to Lys. The uvrD44 has another nucleotide change at +1859, GAA to AAA (Glu at 595 to Lys), which is considered to be the suppressor mutation uvrD44. PMID- 2968514 TI - A new mutagen-sensitive mutant in Neurospora, mus-16. AB - A new gene, mus-16, is determined by the nitrogen mustard-sensitive Neurospora mutant of Baker, Parish and Curtis (1984) which is defective in the removal of DNA-DNA and DNA-protein crosslinks. This gene is on the left arm of linkage group V between caf-1 and lys-1. The mus-16(JMB) mutant is sensitive to the alkylating agents methyl methanesulfonate (MMS) [dose reduction factor (drf) 8-10 X], N methyl-N'-nitro-N-nitrosoguanidine (drf 5-6 X), the amino acid histidine and the drug hydroxyurea. It is not sensitive to ultraviolet-light, gamma-irradiation, or mitomycin C (MMC). It shows normal spontaneous mutation rates but increased induction of mutation by MMS. Homozygous crosses are barren, showing no signs of sporulation. Mitotic spontaneous chromosome instability is increased. The mus-16 mutation is similar to several non-excision repair-defective mutants in Neurospora. Some of these may be defective in repair of alkylation damage. The MMC data supports earlier data that in fungi MMC is incapable of forming DNA-DNA crosslinks. PMID- 2968515 TI - The Ca2+ ATPase content of slow and fast twitch fibers of guinea pig. AB - The Ca ATPase content in the sarcoplasmic reticulum (SR) of fast and slow twitch skeletal fibers was estimated using two well-characterized muscles of the guinea pig: the white bundle of the vastus lateralis and the soleus. Ca ATPase surface density was determined by counting the projections of individual molecules revealed on the cytoplasmic surface of freeze-dried, rotary-shadowed microsomal vesicles isolated from the two muscles. The Ca ATPase densities were 32,000/micron 2 and 25,000/micron 2 for the vastus lateralis and soleus muscles, respectively. The percentage of membrane area occupied by Ca ATPase-free lipid patches was estimated using freeze-fractured, rotary-shadowed in situ SR. In soleus muscle the free SR of terminal cisternae and the longitudinal SR have 34.5 and 19.7% of their surface free of ATPase, respectively. In the white vastus less than 1% of the surface was not occupied by Ca ATPase. These values were combined with stereological data from the literature to give a ratio of total Ca ATPase content per unit fiber volume of 1:2, slow versus fast. This is considerably less than the approximately sixfold difference in the overall relaxation time and the half times to relaxation between the two fiber types. This suggests that other factor such as differences in enzyme kinetics or cytoplasmic Ca buffering proteins must also play a role in determining rate of relaxation. PMID- 2968516 TI - Noncoordinate expression of M band proteins in slow and fast embryonic chick muscles. AB - The expression of the myosin-associated M band proteins myomesin and M protein in differentiating muscle fibers in the anterior latissimus dorsi (ALD) and posterior latissimus dorsi (PLD) muscles during embryonic chicken development was examined by immunocytochemistry using monoclonal antibodies. Early in the embryonic development of both muscles, both myomesin and M protein are expressed in primary and secondary myotubes. However, beginning at 10 days in ovo, M protein is gradually suppressed first in primary, then in secondary, presumptive slow-tonic type 3 fibers. M protein is transiently suppressed in presumptive fast twitch type 2 fibers derived from primary myotubes but continuously expressed in those derived from secondary myotubes. Thus, initially all myotubes have a common intrinsic M band composition with respect to myomesin and M protein, whereas at later stages the expression of M protein is fiber-type specific. Intrafusal spindle fibers, which are segregated from extrafusal fibers around 14 days in ovo, have a heterogeneous M band composition atypical of extrafusal fibers. PMID- 2968517 TI - Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faecium. PMID- 2968518 TI - Transcript localization of four opsin genes in the three visual organs of Drosophila; RH2 is ocellus specific. AB - Drosophila and other Dipteran flies have three different kinds of visual organs; in the adult a pair of compound eyes and three dorsal ocelli; and in the larva a pair of internal photoreceptor organs. They develop in distinct ways, yet have certain features in common. All three organs use retinal-derived chromophores, coupled to distinct opsins, to provide a diversity of spectral sensitivities. Four opsin genes have been identified thus far in Drosophila; Rh1, Rh2, Rh3 and Rh4 (refs 6-11). We have used in situ hybridization to study the messenger RNAs expressed by these four opsin genes in all three visual organs. Rh1, Rh3 and Rh4 are already known to be expressed in different subsets of cells in the compound eye. We found that, in contrast, opsin Rh2 is the predominant opsin expressed in the ocelli. Opsin Rh1 is known to be expressed in the larval photoreceptor. We found that Rh3 and Rh4 are as well, but not Rh2. The ocellar-specific gene expression of Rh2 is of particular interest for its possible bearing on the function of the ocellus. PMID- 2968519 TI - Frontiers of the immune system. PMID- 2968520 TI - Regulatory properties of LFA-1 alpha and beta chains in human T-lymphocyte activation. AB - Lymphocyte function-associated antigen-1 (LFA-1) is a heterodimer composed of an alpha and beta chain that is expressed on the surface of most leukocytes and is an essential molecule for adhesion reactions between cells participating in the immune response. A putative ligand for LFA-1 is the intercellular adhesion molecule ICAM-1 (refs 3-5). Leukocyte adhesion abnormality is found in patients with LFA-1 deficiency. It is not clear whether binding of ligand to the LFA-1 molecule merely spatially orientates cells towards each other or can also induce signals that regulate cell activation and differentiation. We have recently developed a T-cell proliferation assay which uses immobilized anti-CD3 monoclonal antibodies as stimulant and is independent of LFA-1-mediated cellular adhesion. As there is no interference by anti-LFA-1 monoclonal antibodies with the adhesion dependent activation steps, this T-cell activation system allows us to investigate whether transmembrane signals are induced by binding of ligand to LFA 1 on T cells. Our data indicate that binding of ligand to LFA-1 results in the transduction of regulatory signal across the plasma membrane, rather like other molecules (CD2, CD4, CD8) (refs 8-11) with signal-modifying properties involved in the adhesion of T cells to target/stimulator cells. Indeed, adhesion molecules might generally be important in signal transduction, even in cells not belonging to the immune system. PMID- 2968522 TI - Association between an oncogene and an anti-oncogene: the adenovirus E1A proteins bind to the retinoblastoma gene product. AB - One of the cellular targets implicated in the process of transformation by the adenovirus E1A proteins is a 105K cellular protein. Previously, this protein had been shown to form stable protein/protein complexes with the E1A polypeptides but its identity was unknown. Here, we demonstrate that it is the product of the retinoblastoma gene. The interaction between E1A and the retinoblastoma gene product is the first demonstration of a physical link between an oncogene and an anti-oncogene. PMID- 2968521 TI - Expression of the gamma-delta T-cell receptor on intestinal CD8+ intraepithelial lymphocytes. AB - The vast majority of mature T lymphocytes in the peripheral blood and lymphoid organs use the CD3-associated alpha, beta T-cell receptor (TCR) heterodimer for antigen recognition. A second class of TCRs consists of disulphide-linked gamma and delta proteins that are also CD3-associated. A subset of early CD3+ fetal and adult CD4- 8- thymocytes express gamma, delta TCRs before alpha, beta TCRs are detectable. In addition, a minor (1-5%) subpopulation of peripheral T lymphocytes, and some spleen cells from nude mice express gamma, delta TCRs. Notably, dendritic epidermal cells have also been shown to express gamma, delta TCRs. All of these populations lack CD4 and CD8 molecules. We now report that most mature T cells residing in the murine intestinal epithelium express CD3 associated TCRs composed of gamma-chains disulphide-linked to a protein resembling the delta-chain. The striking feature of these intraepithelial lymphocytes (IEL) was that they were exclusively CD4-8+. In addition, approximately half of CD3-bearing IEL lacked detectable Thy-1 on the cell surface, which is unprecedented for murine T cells. In contrast to other CD8+ peripheral T cells, freshly isolated IEL could be induced to display cytolytic activity by engaging the CD3 molecule, indicating that activation had occurred in vivo. Thus, CD8+ IEL are a phenotypically diverse and anatomically restricted population of lymphocytes that use gamma-chain containing heterodimers for antigen recognition. PMID- 2968523 TI - A region of SV40 large T antigen can substitute for a transforming domain of the adenovirus E1A products. AB - SV40 large T antigen contains a small region of amino acid sequence, conserved among the papovaviruses, that shows considerable similarity to conserved domain 2 of the adenovirus E1A oncogene, a domain which plays an important role in the E1A transforming functions. To learn whether the analogous SV40 T antigen sequences could substitute functionally for E1A domain 2, a chimaeric gene was constructed, coding for T antigen amino acid residues 101 to 118 in place of E1A domain 2. The resulting product showed much of the activity of the wild-type E1A products. It induced proliferation of primary BRK cells and cooperated with the ras oncogene to transform these cells fully. In addition, the chimaeric protein coprecipitated two cellular proteins whose specific binding to the E1A products depends on the presence of domain 2. The activity of the chimaeric product suggests that a similar functional unit exists in the transforming proteins of both SV40 and adenovirus, and that these proteins may exert their cell growth regulating effects through similar mechanisms. PMID- 2968524 TI - Free fatty acids, lipid peroxidation, and lysosomal enzymes in experimental focal cerebral ischemia in primates: loss of lysosomal latency by lipid peroxidation. AB - Experimental focal cerebral ischemia was produced in monkeys (Macaca radiata) by occlusion of the right middle cerebral artery (MCA). The release of the lysosomal glycosidases, beta-D-hexosaminidase, alpha-L-fucosidase and alpha-D-mannosidase into the soluble fraction in the right basal ganglia of the experimental animals was measured at different periods from 30 min to 12 hr after occlusion and compared with the corresponding sham operated control animals. There was a significant increase in the released lysosomal enzymes in the MCA occluded animals at all periods and particularly at 4 hr after occlusion. The CSF from the experimental animals also showed elevated levels of hexosaminidase and fucosidase. The free fatty acids (FFA) measured in the basal ganglia at 30 min and 2 hr after occlusion showed a 100 fold increase in the experimental animals. The predominant fatty acid released was linoleic acid (18:2) followed by arachidonic acid (20:4). Lipid peroxidation in the basal ganglia measured by the thiobarbituric acid (TBA) reaction in the presence or absence of ascorbic acid also showed a significant increase in the experimental animals at all periods with a maximum at 30 min to 2 hr after occlusion. In order to assess whether lipid peroxidation causes damage to the lysosomes and release of the enzymes, a lysosome enriched P2 fraction from the normal monkey basal ganglia was prepared and the effect of peroxidation studied. Maximum peroxidation in the P2 fraction was observed in the presence of arachidonic acid, ascorbic acid and Fe2+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968526 TI - [Bloodless evaluation of cardiac morphological parameters in hypertransfused thalassemic patients. Their course throughout time]. PMID- 2968525 TI - Regional processing of the N- and C-terminal domains of proopiomelanocortin in monkey pituitary and brain. AB - The total content and extent of processing of the gamma 3MSH and beta-endorphin containing N- and C-terminal domains of proopiomelanocortin were determined in the anterior and intermediate lobes of the pituitaries and in 11 regions of the brains of three Rhesus monkeys. Most immunoreactive gamma 3MSH and beta-endorphin was located in the pituitary lobes, although significant amounts were also found in several brain regions. Sephadex column chromatography revealed that gamma 3MSH immunoreactivity was found primarily as 4K and 9K forms; no gamma 1MSH was detected. beta-Endorphin immunoreactivity was found as beta-endorphin, beta lipotropin, and as a 5K form which may represent beta-endorphin extended N terminally by part or all of beta-MSH. In the anterior lobe of the pituitary, the predominant products were 9K gamma 3MSH and beta-lipotropin; in the intermediate lobe, more processed forms (4K gamma 3MSH, beta-endorphin and 5K beta-endorphin) appeared to be preferentially stored. The pattern of processing in various brain regions was similar to that of the intermediate lobe of the pituitary. PMID- 2968527 TI - [The role of percutaneous transluminal angioplasty in atherosclerotic lesions of the legs]. PMID- 2968528 TI - [Reliability of Doppler velocimetry with spectrum analysis in the study of vertebral circulation. Correlation with angiographic data on 21 patients]. PMID- 2968529 TI - Community based services for disabled people--a personal opinion. PMID- 2968530 TI - DSSNY to appeal loss in AIDS case. PMID- 2968532 TI - Why physical medicine has gone to the dogs.... PMID- 2968531 TI - Myocardial kinetics of 99Tcm hexakis-(2-methoxy-isobutyl-isonitrile) (HMIBI) in patients with coronary heart disease: a comparative study versus 201Tl with SPECT. AB - In 15 patients with coronary heart disease, 74 MBq 201Tl (A) and 240 MBq 99Tcm HMIBI (B) were injected during exercise on separate days and SPECT imaging was applied. In (B) a consecutive second injection was given with an identical dose 3 h later (at rest) and net rest distribution was obtained from a subtraction technique. From polar, target-like ROIs, data were computed to compare regional vitality, wash-out and wash-out corrected redistribution. Myocardial wash-out, in contrast to 201Tl, was found to be 20% for 99Tcm independent of the regional myocardial state. Since no considerable redistribution occurred, myocardial distribution of 99Tcm HMIBI remained rather constant for 3 h, resulting in a maximum myocardium to lung ration of about 5. After the second injection, redistribution revealed the same results as in the 201Tl targets. From regional exercise to rest ratios a 'perfusion reserve' was determined which was 2.1 in normal and 1.3 in myocardial areas with defects. Myocardial defect uptake and defect size were nearly identical, yielding agreement in 97% of the myocardial segments. We conclude that 99Tcm HMIBI is a most powerful myocardial imaging agent. The final routine protocol, however, is as yet not established. PMID- 2968533 TI - Cell type dependent transformation by adenovirus 5 E1a proteins. AB - The two major proteins of 243 amino acids (243R) and 289R encoded by the E1a region of adenovirus 5 are related, differing only by the presence of an internal conserved 46 amino acid segment in the 289R protein. We report here that these proteins have different cell transformation properties. Primary baby rat kidney (BRK) cells infected with an M-MuLV retrovirus vector that expresses the 243R protein are immortalized and the resulting foci consist predominantly of epithelial cells; a small fraction of the foci consist of fibroblast-like cells. In contrast, BRK cells infected with a retrovirus expressing the 289R protein are immortalized at a very low frequency and the foci induced consist of only fibroblast-like cells. The immortalizing properties of the 289R protein are dramatically changed by certain mutations within the unique 46 amino acid region: M-MuLV vectors with these mutations immortalize epithelial cells with high efficiency, as much as an order of magnitude higher than vectors that express the 243R protein. The enhanced immortalization property of the mutant forms of the 289R protein does not appear to be related to their transcriptional activation function. Our results suggest that the unique region of the 289R protein has the potential to inhibit immortalization of primary epithelial cells. PMID- 2968534 TI - Separation of immortalization and T24-ras oncogene cooperative functions of adenovirus E1a. AB - An adenovirus 2 E1a gene coding for a protein of 243 (243R) amino acids can efficiently immortalize primary rat kidney (BRK) cells and cooperate with the activated cellular ras oncogene (T24 ras). A mutant (47-0) of the 243R gene that maps between amino acid residues 47-50 within a region that is highly conserved among the various adenovirus serotypes was found to be severely defective in immortalization. Despite the defect in immortalization, mutant 47-0 had the ability to cooperate with T24 ras in oncogenic transformation. These results suggest that the immortalization and the oncogene cooperation functions of the 243R are separable. Our results further suggest that the requirement for a separate immortalization function can be circumvented by oncogenic transformation and that the immortalization of cells transformed by E1a and T24 ras may be a secondary consequence of transformation by these two oncogenes. PMID- 2968535 TI - [Losses cause by ixodid and demodectic ticks and mites in the tanning industry]. AB - Damage done to skin of cattle and defects of raw leather, which are caused by ixodid and demodecid ticks, parasitic on animals, are described. Losses of tanning industry amount to 1.6 to 3.7 thousand roubles per each thousand of finished box-calf leathers. PMID- 2968536 TI - [Communication in the trisomy 21 patient: observation and therapeutic possibilities]. AB - The behaviour of 4- to 6-year-old children with Down syndrome was studied by audio-visual means during individual or group stimulation sessions. From these observations, it is concluded that children with Down syndrome have a rich preverbal communication, but poor child-to-child interchanges. Thus, when taking care of children with Down syndrome, it is necessary to accentuate individual stimulation and take into account the affective drive of the child towards the adult. PMID- 2968537 TI - [A case report of Budd-Chiari syndrome treated with PTA--usefulness of a peripheral atherectomy catheter]. PMID- 2968538 TI - Identification of a mouse homolog of the human laminin receptor. PMID- 2968539 TI - Sequence of bacteriophage Mu N and P genes. PMID- 2968540 TI - Determination of the molecular weight of DNA-bound protein(s) responsible for gel electrophoretic mobility shift of linear DNA fragments examplified with purified viral myb protein. AB - A protein-DNA complex has less gel electrophoretic mobility than the free DNA fragment. One parameter for the degree of retardation of a linear DNA fragment in a protein-DNA complex is the molecular weight of the bound protein(s). The quotient of the migration distances of free DNA (m) and protein-DNA complex (m') is a function of the molecular weight (MW) of the bound protein(s). Based on the evaluation of the lac repressor induced mobility shift of a 203 bp DNA fragment containing the lac operator in a 5% non-denaturating polyacrylamide gel a direct proportionality could be shown between (m/m'-1) and MW with the proportionality factor K = 215 kDa. The factor K depends on the acrylamide concentration in the gel, getting lower values with increasing acrylamide concentrations. A calculation is given to determine the molecular weight of DNA-binding factors responsible for the decreased electrophoretic mobility of a linear DNA fragment. As an example this calculation was used in order to analyse DNA-binding of the isolated viral myb protein. It could be demonstrated that the viral myb protein binds to DNA as a monomer and as a dimer. PMID- 2968542 TI - Dead to the world. PMID- 2968541 TI - Microinjected deoxynucleotides for the study of chemical inhibition of DNA synthesis. AB - Microinjection is shown to be a useful tool for studies of chemical inhibition of DNA synthesis: inhibitor-treated cells were injected with combinations of radioactive precursors and their uptake into DNA was monitored by autoradiography. The results obtained from inhibition by cytosinearabinoside, aphidicolin, trifluorothymidine, and fluorodeoxyuridine agreed well with the common knowledge about these drugs. Short-term (but not long-term) treatments with methotrexate were compensated by injections of thymidine-nucleotides. The effect of hydroxyurea was in part, but not fully, reversed by injection of all four deoxytriphosphates; this implies a second mechanism besides inhibition of ribonucleotide reductase. Regulation of reductase was responsible for the effect of thymidine: the enhanced dTTP caused a depletion of dCTP and dATP. Novobiocin was different from all other drugs tested, DNA polymerase or enzymes of the precursor metabolism are obviously not targets of this drug. PMID- 2968543 TI - [Hypersensitivity to sulfonamides: its incidence and clinical picture]. PMID- 2968544 TI - Markers for neuroendocrine differentiation. AB - Several morphological methods are currently available for the demonstration of neuroendocrine differentiation. Silver impregnation techniques to show argyrophilia or argentaffinity are reliable albeit limited tests for neuroendocrine cells. Immunocytochemistry, at the light or electron microscopic level, is now a routine technique in many histopathological departments. General neuroendocrine markers can be immunostained to define the nature of a tumour or, using antisera to specific neuroendocrine cell products, the exact type of tumour can be established. The application of a range of antisera to bioactive peptides and other derivatives of their precursors can increase the efficiency of detection and classification of neuroendocrine tumours. The latest technology available goes beyond looking at neuroendocrine cell products and investigates peptide gene expression. In situ hybridisation of DNA or mRNA species thereby adds a new dimension to the morphological analysis of neuroendocrine tumours. Used in combination with immunocytochemistry, this technique can provide information on intracellular hormone synthesis. The study of markers of neuroendocrine differentiation continues to yield information which aids not only the diagnosis of neuroendocrine tumours but also the understanding of their biology and the effects exerted by their products. PMID- 2968545 TI - Immunoprophylaxis of viral hepatitis. AB - Because safe, effective treatment for established viral hepatitis is not available, physicians need to be acquainted with recent advances in prophylaxis. Immune globulin (Gamastan, Gammar) is used for both preexposure and postexposure prophylaxis of hepatitis A, and side effects are rare. Two vaccines (Heptavax-B, Recombivax HB) are licensed for hepatitis B, one is a plasma-derived vaccine, the other a yeast-recombinant vaccine. Indirectly, these also control hepatitis D (delta agent), which needs hepatitis B virus to develop. PMID- 2968546 TI - [Hairy cell leukemia: therapeutic strategy]. PMID- 2968547 TI - [Epilepsy: indications for the surgical treatment]. PMID- 2968549 TI - [Treatment of post-neurosurgical diabetes insipidus with desmopressin by intramuscular route]. AB - Desmopressin (DDAVP) is used intramuscularly in the treatment of post operative diabetes insipidus as soon as the condition is diagnosed to ensure continuous replacement of antidiuretic hormone secretion during the first 5 days of therapy. Two successive studies, each involving 15 patients, were conducted. The first study was designed to test the effectiveness and detect the possible side effects of intramuscular DDAVP, while the purpose of the second study was to evaluate the clinical application of the drug. With seven 2 mcg doses of DDAVP, administered 12-hourly by intramuscular injection to patients weighing more than 30 kg, continuous antidiuresis during 96 hours was achieved. This method is simple and effective, but it should not be prolonged beyond that period of time. Moreover, to prevent plasma hypo-osmolality, fluid intake must be strictly controlled and kept at the same level as or below diuresis. PMID- 2968548 TI - [Systematic inhibition of the luteinizing hormone with a gonadoliberin analog, triptorelin, during ovarian stimulation for fertilization in vitro. Choice of protocol]. AB - The quality of ovarian stimulation for in vitro fertilization with or without an LH-RH analogue was investigated in a randomized trial involving 30 women divided into 3 groups. Group I women were treated with the conventional clomiphene citrate-human menopausal gonadotropin combination without LH-RH analogue. Group II women (long regimen) received a slow-release preparation of triptorelin (DTRp6 LH-RH), an LH-RH analogue, and human menopausal gonadotropin. Group II women (short regimen) were given triptorelin with human menopausal gonadotropin. Inhibition of the endogenous luteinizing hormone using triptorelin improved the results of in vitro fertilization in group II and group III women, but the short regimen was distinctly less compelling and less expensive than the long regimen. PMID- 2968550 TI - [Decrease of erythrocyte receptors for the C3b fragment of complement in acquired immunodeficiency syndrome]. AB - The number of C3b receptors (CR1) on erythrocytes (E) was measured, using a newly developed flow cytometry assay and a standard radioimmunoassay, in HIV-infected individuals and in a normal control population. The number of CR1/E was significantly decreased in symptomatic HIV-infected patients with ARC and acquired immunodeficiency syndrome and was normal in asymptomatic carriers of anti-HIV antibodies. Decreased numbers of CR1/E correlated with the clinical severity of the HIV-related disease. A close correlation was found between results obtained with flow cytometry and those obtained with the radioimmuno assay. Flow cytometric assessment of CR1/E may be of interest in large scale prospective clinical studies of HIV-infected subjects. PMID- 2968551 TI - [Value of the renal graft as a function of blood creatinine of the donors]. AB - In a series of 851 renal transplantations, the fate of the grafted kidneys was studied in relation to the donors' renal function. The actuarial survival curves of the grafts showed no significant difference at 24 months between donors with blood creatinine levels below or above 150 mumol.l-1. Seventy-five of the 99 kidneys obtained from donors with high blood creatinine were functioning at 2 years. It would appear that the classical doubts about renal procurement from donors with high blood creatinine levels should be reconsidered. PMID- 2968552 TI - [Significance of antimitochondrial antibodies. In a population of 111 patients from the Midi-Pyrenees region]. AB - The aetiological significance of antimitochondrial antibodies (anti-M) being debated, we conducted an aetiological study of 111 sera with anti-M2 antibodies from patients living in the Midi-Pyrenees region (South-West France); these anti M2 antibodies had been detected in the years 1984-85. Fifty-nine percent of the patients had primary biliary cirrhosis (41 p. 100 were asymptomatic, and 6 p. 100 had normal liver function tests). Anti-M2 antibodies at a titer of 1/500 or more were significantly more frequent among these patients as compared with the others (P less than 0.001, probability = 0.73). Six percent of the patients had chronic active hepatitis. The incidence of primary biliary cirrhosis could be estimated at 8.5 per 10(6) people per annum. In this patient population, primary biliary cirrhosis was more common than all other causes of anti-M2 antibodies, and an anti-M2 titer of 1/500 or more had a 73 p. 100 predictive value for that disease. In the region concerned the incidence of primary biliary disease was higher than previously reported in Western Europe. PMID- 2968553 TI - [First-line treatment of cardiac insufficiency: digitalis glycosides, diuretics, vasodilator agents or new cardiotonic agents?]. AB - The advent of vasodilator therapy and new cardiotonic agents has modified the medical treatment of heart failure. First-line treatment is no longer standard; its choice must take into account a number of individual factors such as patient's age, causative cardiopathy, severity of the disease, response to treatment, contra-indications and side-effects of drugs. The aim is to obtain an improvement not only in the patient's comfort but also in long-term prognosis. The prescription of digitalis compounds to heart failure patients with sinus rhythm has been questioned. Alternatives to the conventional therapeutic approach based on cardiac glycosides and diuretics are now available. They include the use at an early stage of vasodilator therapy, and notably angiotensin-converting enzyme inhibitors, in the hope of avoiding gradual reduction of cardiac performance. PMID- 2968554 TI - [Diagnosis of gastroesophageal reflux. Comparison of long-term pHmetry and 3-hour postprandial pHmetry]. PMID- 2968555 TI - [Escherichia coli pelviperitonitis associated with pneumoperitoneum]. PMID- 2968556 TI - [Neonatal herpetic laryngitis. Value of early diagnosis]. PMID- 2968557 TI - [Major ST segment elevation after cardioversion]. PMID- 2968558 TI - [Persistence and severity of post-transfusion hepatitis caused by hepatitis B virus]. PMID- 2968559 TI - [Value of noradrenaline in the treatment of hyperkinetic septic shock]. PMID- 2968560 TI - [Interaction of lithium and angiotensin-converting enzyme inhibitors]. PMID- 2968561 TI - [Popliteal venous aneurysm]. PMID- 2968562 TI - [Transfusion malaria caused by Plasmodium ovale. A case]. PMID- 2968563 TI - [Role of the CR1 receptor for complement C3b fragment]. PMID- 2968564 TI - [High doses of calcium carbonate or the combination of 1 alpha hydroxylated derivatives of vitamin D and aluminum compounds bound to phosphorus in the preventive treatment of renal osteodystrophy]. PMID- 2968565 TI - [Respective frequency and radioclinical features of 150 lung diseases observed in 125 patients with human immunodeficiency virus infection]. AB - Seventy out of 125 patients with HIV infection had diffuse alveolo-interstitial pneumonia usually caused by an opportunistic infection, notably pneumocystosis. Nineteen patients had only localized lung opacities due either to usual or tuberculous bacterial infections or to Kaposi's sarcoma. In 10 patients with pleural effusion or mediastinal adenopathy, the condition was due to Kaposi's sarcoma (n = 4) or to mycobacteriosis (n = 3). An opportunistic or usual infection was demonstrated in 17 of the 51 patients with normal radiography of the chest. Finally, 37 patients free from infectious or tumoral pathology had isolated lymphocytic alveolitis. PMID- 2968566 TI - [Systematic ultrasensitive determination of thyroid-stimulating hormone in 50 cases of atrial fibrillation]. AB - The incidence of hyperthyroidism in patients with continuous arrhythmia due to atrial fibrillation was evaluated prospectively by means of the "ultrasensitive" assay of thyroid-stimulating hormone (TSH). When the result was abnormal, free thyroxine fractions (FT4) and free triiodothyronine fractions (FT3) were measured, and in some cases a thyrotropin-releasing hormone (TRH) test and a thyroid gland scintigraphy were performed. Among 50 unselected patients with continuous arrhythmia due to atrial fibrillation, 12 had a low TSH baseline level. In 7 of them, high FT4 and/or FT3 levels provided evidence of hyperthyroidism. In the remaining 5 patients FT4 and FT3 levels were normal, but TSH response to TRH was suppressed and scintigraphy showed a high nodular or lobular uptake. In a control series of 50 age- and sex- matched subjects without thyroid disorders or dysrhythmia, TSH level was low in only one case with insufficient response to TRH, but the scintigraphic image did not suggest nodular hyperthyroidism. It is concluded that systematic TSH assays can detect those atrial fibrillations which are consecutive to, or aggravated by hyperthyroidism, even asymptomatic. We regard this assay as indispensable to evaluate continuous arrhythmia due to atrial fibrillation, even when an apparently causative underlying heart disease is present. PMID- 2968568 TI - [French, European and international congresses, medical and scientific meetings from 1988 to 1989 inclusive. Complementary information]. PMID- 2968567 TI - [Allergy to mercurothiolate in an infant during heparinization of an intracaval catheter]. AB - A 4-month old infant developed an immediate and proven systemic allergic reaction to mercurothiolate. The acute accident occurred while an intracaval catheter was being treated with a dry-frozen heparin which excipient contains mercurothiolate. This conservative agent is present in numerous pharmaceutical preparations for topical and systemic use. PMID- 2968569 TI - [Progressive dystonia and putamen lesions in magnetic resonance imaging]. PMID- 2968570 TI - [Benign teratoma of the thyroid in one adult]. PMID- 2968571 TI - [Prolonged remissions in type I diabetes mellitus with ketosis]. PMID- 2968572 TI - [Myoglobin release in myocardial ischemia: a cause-effect relationship?]. PMID- 2968573 TI - [Hyperammonemia coma following uretero-sigmoidostomy]. PMID- 2968574 TI - [Panlobular emphysema in the terminal stage: unilateral pulmonary graft]. PMID- 2968575 TI - [Benign tumors of the stomach. Value of echo-endoscopy for diagnosis and surgical indications]. PMID- 2968576 TI - [Myocardial infarction in a patient with Pendred syndrome]. PMID- 2968577 TI - [Threatening hyperkalemia caused by mannitol. A little-known side effect]. PMID- 2968578 TI - [Research approach in the diagnosis and treatment of neuroblastoma]. PMID- 2968579 TI - [Home parenteral nutrition for adults. Results of a multicenter survey in France]. AB - Home parenteral nutrition (HPN) is a growing therapy in North America and Europe. This first multicenter retrospective study of HPN in France has collected data on 81 patients sent home before December 31st, 1985. Intestinal failure, secondary to short bowel syndrome, small bowel stenosis or fistula, was the main indication for HPN. In 95 p. 100 of the cases, the clinical nutritional status during HPN was either normal or subnormal. The annual incidence of catheter change for technical complication was 0.78 and the mortality rate was 1.2 p. 100. Social rehabilitation was recovered during HPN in 60 p. 100 of patients. Thirty percent of the patients died of their primary disease during HPN but 43 p. 100 were off treatment, and 27 p. 100 were on HPN at the end of the study. The cost of HPN was reduced by 64 p. 100 in comparison with the cost of parenteral nutrition carried out in hospital. PMID- 2968580 TI - [Bronchogenic esophageal cysts in the adult. Four cases]. AB - Bronchogenic oesophageal cysts possess a mucosa of the airway type but are unconnected to the tracheobronchial tree; they are in close anatomical relation with the oesophageal wall. These characteristics are explained by the fact that the respiratory and digestive tract share the same embryonic development. Four cases of bronchogenic oesophageal cyst are reported, and 21 well-documented cases from the literature are reviewed. The lesion is extremely rare in adults, often complicated and responsible for gastrointestinal, respiratory and sometimes even cardiac symptoms. Its features at radiology, endoscopy and computerized tomography are suggestive of the diagnosis. Treatment consists of excision by enucleation. Excision must be complete for recurrences to be avoided. PMID- 2968581 TI - [Hand-Schuller-Christian disease with tumor localization in the posterior fossa]. AB - A 19-year-old girl had been suffering since the age of 4 from Hand-Schuller Christian disease. The condition was diagnosed by biopsy of a cranial bone lacuna. Later on, she developed diabetes insipidus and bilateral exophthalmos, both being typical signs of Hand-Schuller-Christian disease. At the age of 14, an intracranial tumour in the posterior cerebral fossa was discovered by computerized tomography, but in view of her parent's refusal of surgery, she was operated upon only 4 years later for obstructive hydrocephalus with intracranial hypertension. Two years after this operation, she presented with neurological signs of compression of the medulla oblongata, including tetraparesis and respiratory disorders, and the tumour was resected. Following a brief improvement she died of major hydroelectrolytic disorders due to her irreducible unstable diabetes insipidus. A review of the literature on Hand-Schuller-Christian disease and its rare association with intracranial tumours is analysed. The occurrence of such tumours raise important therapeutic problems and make the prognosis considerably worse. PMID- 2968583 TI - [Pregnancy and labor in a heart transplant patient]. PMID- 2968582 TI - [High-density cholesterol lipoprotein: metabolism and role in atherosclerosis]. AB - High density lipoproteins (HDL), also called alpha lipoprotein, are small high protein content particles that are present in the vascular and extravascular systems. Two main classes of HDL, HDL2 and HDL3, have been identified and found to have different structures and different metabolisms. Their principal function is to ensure that tissue cholesterol returns to the liver. HDL, and notably HDL2, are factors that protect against cardiovascular diseases. HDL2 are subjected to the influence of numerous dietetic, genetic and hormonal factors, while HDL3 seem to be more independent of all variations, except those of alcohol. PMID- 2968584 TI - [Anuria caused by bilateral ureteral stenosis in Wegener's granulomatosis]. PMID- 2968585 TI - [Right intraventricular thrombosis and thrombopenia due to heparin]. PMID- 2968586 TI - [Visceral leishmaniasis in acquired immunodeficiency syndrome]. PMID- 2968587 TI - [Parallel increase in the prevalence of pulmonary tuberculosis and infection by human immunodeficiency virus in Bangui]. PMID- 2968588 TI - [Rare cause of compression of the terminal conus: osteoporosis]. PMID- 2968589 TI - [Lupus syndrome induced by application of a dermatological]. PMID- 2968590 TI - [Nocturnal mechanical ventilation in intermittent positive pressure at home by nasal route in chronic restrictive respiratory insufficiency. An effective substitute for tracheotomy]. PMID- 2968591 TI - [Intracranial subdural hematoma in the period following spinal anesthesia]. PMID- 2968592 TI - [Hormonal characteristics of interstitial tumors of the testis secreting estrogens]. PMID- 2968593 TI - [Indications for the laser in digestive tumor pathology]. PMID- 2968594 TI - [Value of the urinary albumin/creatinine ratio in the detection of microalbuminuria]. AB - Microalbuminuria is currently defined as a urinary albumin excretion rate of 20 to 200 micrograms per minute, measured in the urine of 24 hours. We present an indirect approach to the urinary albumin daily excretion rate which minimizes the errors due to chronometric measurement and to the difficulty of collecting 24 hour urine. The albumin (mg/l)/creatinine (mmol/l) ratio calculated in urine collected daily indicates microalbuminuria when it is higher than 2.97 in women and 2.48 in men. The interpretation of this ratio is discussed in terms of sensitivity, specificity and predictive value. PMID- 2968595 TI - [Effects of simvastatin on plasma lipids, lipoproteins and apoproteins (A1 and B). 24 cases of major primary hypercholesterolemia]. AB - We studied the effects of simvastatin (MK 733), a new competitive inhibitor of HMG CoA reductase, alone and in combination with a bile acid sequestrant, cholestyramine, on serum levels of lipoproteins and apoproteins A1 and B, in 24 patients with familial hypercholesterolemia. After simvastatin treatment (40 mg/day) alone for 12 weeks, serum total and low density lipoprotein cholesterol decreased by 31 and 36 percent respectively. With the addition of cholestyramine, there was a 41 per cent total decrease in serum cholesterol from the control value and a 50 percent decrease in low density lipoprotein cholesterol. After cholestyramine treatment alone for 12 weeks, serum total and low density lipoprotein cholesterol decreased by 20 percent and 29 percent respectively. With the addition of simvastatin (20 mg per day), there was a 32 percent total decrease in serum cholesterol from the control value and a 43 percent decrease in low density lipoprotein cholesterol. High density lipoprotein cholesterol remained unchanged. No major adverse effect was observed. If long term safety can be confirmed, the simvastatin-cholestyramine regimen may prove useful in heterozygous familial hypercholesterolemia. PMID- 2968597 TI - [Study of the circle of Willis with digital angiography. Contribution to the detection of high-risk carotid clamping]. AB - Radiological studies of Willis' circle morphology are mainly performed in search of intracerebral aneurysms, and for this purpose digital imaging has not superseded conventional radiology. In contrast, conventional imaging does not seem to have given satisfactory results in the functional study of this substitute vascular network, and it is in this field that digital angiography is of particular value. Between January, 1985 and June, 1986, we performed 300 digital angiographies of the supra-aortic vessels in patients with carotid artery stenosis. Each exploration included a stage of investigation for intracranial substitute vessels by compression of the carotid artery on the side of the lesion. Substitute vessels were found to be adequate in 93 p. 100 of the cases. The test showed that there was no alternative blood flow in 3.7 p. 100 of the cases, which made it imperative to use brain protection measures during clamping of the carotid artery. The substitute network was found to be insufficient in 1 p. 100 of the cases, and the method failed in 2.4 p. 100. PMID- 2968596 TI - [Diagnostic value of a distinct rise in sedimentation rate]. AB - Among the 931 patients who were admitted, over a 9-month period, to an internal medicine department, a group of 84 patients (9 p. 100) whose erythrocyte sedimentation rate (ESR) was 70 mm or more at 1 hour was selected and compared to the remaining 847 patients whose ESR was below 70 mm at 1 hour. In most cases, a pathology likely to account for the distinct rise observed in ESR was found (infection in 42 p. 100 of the cases, malignant disease in 27 p. 100, inflammation in 20 p. 100), and only 5 p. 100 of these rises remained unexplained. This makes an ESR of 70 mm or more a good index of morbidity generally, without pointing at any specific disease. An ESR of 70 mm or more has very low sensitivity (always below 30 p. 100), so that no disease whatsoever can be excluded when the ESR is only slightly elevated. Moreover, in all but infectious diseases a distinctly high ESR is not an index of severity. PMID- 2968598 TI - [Aging of oxygen extractors]. PMID- 2968599 TI - [Dysautonomic orthostatic hypotension in human immunodeficiency virus infection. Remission in azidovudine therapy]. PMID- 2968600 TI - [Assay of 3-methoxy-4-hydroxyphenylethylene glycol in hypertensive outbreaks after cardiac surgery. Effect of clonidine]. PMID- 2968601 TI - [Hypomanic attack under slow-release theophylline. A case report]. PMID- 2968602 TI - [Inappropriate antidiuretic hormone secretion syndrome caused by Horton's disease]. PMID- 2968603 TI - [A case of acquired anti-factor V circulating anticoagulant]. PMID- 2968604 TI - [Shock liver...without shock]. PMID- 2968605 TI - [Effect of concomitant alcoholism on the outcome of tuberculosis]. PMID- 2968606 TI - Chitin synthase 2 is essential for septum formation and cell division in Saccharomyces cerevisiae. AB - Previous work led to the puzzling conclusion that chitin synthase 1, the major chitin synthase activity in Saccharomyces cerevisiae, is not required for synthesis of the chitinous primary septum. The mechanism of in vivo synthesis of chitin has now been clarified by cloning the structural gene for the newly found chitin synthase 2, a relatively minor activity in yeast. Disruption of the chitin synthase 2 gene results in the loss of well-defined septa and in growth arrest, establishing that the gene product is essential for both septum formation and cell division. PMID- 2968607 TI - Molecular cloning of the common acute lymphoblastic leukemia antigen (CALLA) identifies a type II integral membrane protein. AB - Common acute lymphoblastic leukemia antigen (CALLA) is a 100-kDa cell-surface glycoprotein expressed on most acute lymphoblastic leukemias and certain other immature lymphoid malignancies and on normal lymphoid progenitors. The latter are either uncommitted to B- or T-cell lineage or committed to only the earliest stages of B- or T-lymphocyte maturation. To elucidate to homogeneity, obtained the NH2-terminal sequence from both the intact protein and derived tryptic and V8 protease peptides and isolated CALLA cDNAs from a Nalm-6 cell line lambda gt10 library using redundant oligonucleotide probes. The CALLA cDNA sequence predicts a 750-amino acid integral membrane protein with a single 24-amino acid hydrophobic segment that could function as both a transmembrane region and a signal peptide. The COOH-terminal 700 amino acids, including six potential N linked glycosylation sites compose the extracellular protein segment, whereas the 25 NH2-terminal amino acids remaining after cleavage of the initiation methionine form the cytoplasmic tail. CALLA+ cells contain CALLA transcripts of 2.7 to 5.7 kilobases with the major 5.7- and 3.7-kilobase mRNAs being preferentially expressed in specific cell types. PMID- 2968608 TI - Wheat germ agglutinin-positive cells in a stem cell-enriched fraction of mouse bone marrow have potent natural suppressor activity. AB - In the present study we have characterized natural suppressor (NS) cells, which nonspecifically suppress mitogen responses and mixed-lymphocyte reaction. The strongest NS activity was found in a fraction of relatively low-density cells (1.063 less than p less than 1.075) obtained by equilibrium density centrifugation. Further purification and characterization of these NS cells by using a fluorescence-activated cell sorter indicated that wheat germ agglutinin (WGA)-positive cells have potent NS activity, whereas WGA-negative cells have no NS activity. Spleen colony-forming unit (CFU-S) assays demonstrated a significant correlation between the number of CFU-S cells and the NS activity in the bone marrow. However, WGA-positive cells obtained from the bone marrow of animals treated with 5-fluorouracil, which cells in turn were exposed to 5-fluorouracil, showed a marked reduction in NS activity. These results suggest that the pluripotent stem cells have NS activity when the cells are in the cycling phase but not when the cells are in the G0 phase. It seems possible that apparently primitive cells play an important role in down-regulation of immune responses. PMID- 2968609 TI - Incidence of diastasis recti abdominis during the childbearing year. AB - This study was conducted to determine 1) the incidence of diastasis recti abdominis among women during the childbearing year and 2) the location of the condition along the linea alba. Clinicians have long noted its presence, prenatally and postnatally, but the magnitude of the problem is currently unknown. A cross-sectional design was used to test 71 primiparous women placed in one of five groups, based on placement within the childbearing year. A commonly accepted test for diastasis recti abdominis was performed. Palpation for diastasis recti abdominis at the linea alba was performed 4.5 cm above, 4.5 cm below, and at the umbilicus. Diastases were observed at all three places, but most often at the umbilicus. A significant relationship (p less than .05) was found between a woman's placement in her childbearing year and the presence or absence of the condition. Diastasis recti abdominis was observed initially in the women in the second trimester group. Its incidence peaked in the third trimester group; remained high in the women in the immediate postpartum group; and declined, but did not disappear, in the later postpartum group. These findings demonstrate the importance of testing for diastasis recti abdominis above, below, and at the umbilicus throughout and after the childbearing year. PMID- 2968611 TI - Hepatitis B vaccination for staff in institutions for the mentally handicapped. PMID- 2968610 TI - Effects of medroxyprogesterone acetate on plasma testosterone and sexual behavior in male cynomolgus monkeys (Macaca fascicularis). AB - Medroxyprogesterone acetate (MPA), a synthetic progestin with androgen-depleting activity, is used to treat the deviant sexual behavior of men. To investigate the effects of MPA in another anthropoid primate, 16 oppositely-sexed pairs of cynomolgus monkeys were observed in one-hour behavior tests during 15 successive 4-week periods conducted before, during and after administering to males weekly IM injections of first 20 mg and then 40 mg MPA. The doses used were comparable on a body weight basis to those employed clinically. Blood samples were collected weekly and assayed for plasma testosterone. During MPA treatment both plasma testosterone and ejaculatory behavior were significantly decreased, but the changes in behavior were less marked than the changes in hormone levels. There were clear differences between individual males in the effects of treatment, and the identity and hormonal status of the female partners also influenced the results. During the 6 month withdrawal period, effects were only partially reversible, and the data suggested that the behavioral changes depended on the hormonal changes rather than the opposite. However, a direct central action of MPA on behavioral mechanisms could not be excluded. PMID- 2968612 TI - Legal case briefs for nurses. MO.: falsifying schedule: dismissal results; MT.: talc reaction: workers' compensation. PMID- 2968613 TI - The dopamine D2 receptor agonists, quinpirole and bromocriptine produce conditioned place preferences. AB - 1. The conditioned place preference paradigm was used to examine the role of the D2 receptor in mediating the reinforcing effects of dopamine (DA) agonists. 2. During the 3-day pre-exposure phase, rats explored two distinctive end compartments which were adjoined by a small tunnel. During the 8-day conditioning phase, groups of rats were treated with the selective D2 receptor agonists, quinpirole (0.01, 0.025, 0.05, 0.10, 0.25, 1.0 and 5.0 mg/kg IP) or bromocriptine (0, 0.01, 0.1, 0.5, 1.0, 5.0 and 10.0 mg/kg IP) and confined to one compartment for 30 min. On alternate days, rats received vehicle injections and were placed in the opposite compartment. Test days occurred over the remaining 3 days during which untreated animals explored both compartments. 3. Rats conditioned with quinpirole or bromocriptine showed significant increases in time spent in the drug-paired environment from pre-exposure to test indicating the establishment of conditioned place preferences. 4. This suggests a functional role for the D2 receptor in mediating the rewarding effects of DA agonists. PMID- 2968614 TI - [Skin changes associated with HIV infection]. PMID- 2968615 TI - [13-cis-retinoic acid in the treatment of severe forms of acne]. PMID- 2968616 TI - [Clinical course of juvenile acne and the serum levels of zinc in patients treated with Zincteral-Polfa]. PMID- 2968617 TI - [Centenary of the use of contact lenses]. PMID- 2968618 TI - [Traumatic ocular syndrome: the emergency]. PMID- 2968619 TI - [Correlations between kinetic visual acuity and a battery of psychological tests. The psychological values of the tests]. PMID- 2968620 TI - [Detection of color vision disorders in chronic hepatopathies by using pseudoisochromatic charts]. PMID- 2968621 TI - [Eye diseases caused by dust]. PMID- 2968622 TI - [Treatment of ocular herpes with acyclovir]. PMID- 2968623 TI - [Rare case of posttraumatic ciliochoroidal detachment with an original surgical solution]. PMID- 2968624 TI - [Luxation of a prepupillary artificial lens into the posterior chamber and repositioning of the lens]. PMID- 2968625 TI - [Arguments for the nonabusive use of topical chloramphenicol in ophthalmology]. PMID- 2968626 TI - [Ischemic optic neuropathy in Horton's disease]. PMID- 2968627 TI - [Medical complications in the surgery of senile cataract]. PMID- 2968628 TI - [Birdshot retinochoroidopathy]. PMID- 2968629 TI - [Clinical forms of aniridia and associated eye lesions in the same family]. PMID- 2968630 TI - [Angiospastic retinopathy in hypertension with renal sclerosis]. PMID- 2968631 TI - [Pigmentary retinopathy--a disease of an immunologic nature?]. PMID- 2968632 TI - Occupational hazards and low back pain. AB - In a cross-sectional study of 2684 males from a range of occupations the prevalence rate for back pain was 20% (11% disc disease and 9% undetermined pain) and about half of these had pain related to the lower back with the remainder being located in the dorsoscapular or cervical regions. Follow up study of a cohort of 1249 of these workers over a two year period indicated that the incidence rate of low back pain in that group was about 1% per annum. When sickness absence certificates were studied the annual absence from back pain (all sites) was 1323 days per thousand employees. However, when confirmation was obtained from general practitioners about non-specific rheumatic diagnoses this figure rose to 1707 days. The prevalence of disc disease is higher among those engaged in heavy rather than among those in light work, and also among those required to adopt a chronic stooping posture at the place of work. A device designed to record lumbar EMG, antero-posterior and lateral posture of the lumbar spine and intra-abdominal pressure designed to be used in work places over a complete shift is described and traces obtained under laboratory conditions are presented. PMID- 2968633 TI - A molecular genetic approach to Huntington's disease. AB - At this time it is not possible to predict exactly how the function of the HD gene will be determined even after the responsible DNA mutation has been identified. The discovery of a function for a protein or gene of known structure is still a major problem in all species. The ease or difficulty of performing this step in HD will depend in large part on the nature of the HD gene. Characterization of the HD gene and its normal counterpart will certainly provide improved diagnostic capability. It is to be hoped that this information will also lead to effective therapy to ease the suffering of the disease's victims. In the long term, investigations of this type in HD and other neurogenetic disorders will undoubtedly provide new insights into the operation of the human nervous system. PMID- 2968634 TI - Phage typing coagulase-positive staphylococci from rooks and gulls. AB - Phage typing was performed on 86 strains of Staphylococcus aureus and 25 strains of Staphylococcus intermedius from rooks and gulls with human, bovine, chicken and canine phages. Eighty per cent of the S aureus strains and 64 per cent of the S intermedius strains were typable. The S aureus biotype D strains of rook origin were specifically lysed at routine test dilution (RTD) by chicken phages from groups I or I + IV, by human phages belonging to groups I and M, and partly by canine phage 58. The other rook and gull S aureus strains did not show characteristic phage patterns. The S intermedius strains isolated from both species of birds could be typed only with canine phages and this correlated with their classification into biotypes. All the biotype 1 strains tested but only two of 12 biotype 2 strains were lysed with canine phages at RTD. PMID- 2968635 TI - [Clinical evaluation on the response of plasma atrial natriuretic peptide to exercise in healthy volunteers]. PMID- 2968636 TI - [Drug treatment of lumbago and sciatica]. PMID- 2968637 TI - [Therapeutic indications in lumbago]. PMID- 2968638 TI - [Rehabilitation in lumbago and sciatica]. PMID- 2968639 TI - [Lumbago and sciatica. Anatomy and physiopathology]. PMID- 2968640 TI - [Clinical and radiologic symptomatology. Requesting complementary examinations. Which ones and when?]. PMID- 2968641 TI - [Movements of the lumbar spine. A biomechanical study]. AB - The lumbar spine presents movements of flexion-extension, lateral inflexions and rotations, which are, in fact, the result of movements within each mobile spinal segment. These mobile segments present six levels of freedom corresponding to translation and rotation movements in the three spacial planes. Lumbar mobility decreases markedly with age. Muscles not only participate in the movement itself, but also in its control and in maintaining spinal stability. Their action also depends on the effect of gravity. The authors report various studies concerning spinal stresses during varied movements and positions. Flexion of the trunk is the most stressing; walking, rotations and lateral inflexions increase only moderately the intradiscal pressure. All of the these results concur with the classic ergonomic notions. PMID- 2968642 TI - [Spinal pathology in drivers of public transports]. PMID- 2968644 TI - [Biomechanics of the lumbar spine and sailing]. PMID- 2968643 TI - [Lumbar biomechanics and sports. Spondylolysis of L5]. PMID- 2968645 TI - [Biomechanics of the lumbar spine and posture training]. PMID- 2968647 TI - Musculoskeletal disorders and disability in Finland. PMID- 2968648 TI - [Parathyroid crisis. Fatal course despite control of hypercalcemia with mithramycin]. AB - A 73-year-old patient suddenly became confused and comatose due to excessive hypercalcemia. He developed acute respiratory distress syndrome (ARDS) and died even though the hypercalcemic state had been brought under control with mithramycin. Autopsy disclosed an adenoma of the parathyroid gland and calcifications in lung and kidney tissues as well as hyaline membranes. The case illustrates the poor prognosis of hyperparathyroid crisis whenever organ damage is present. PMID- 2968646 TI - Hormonal effects of high dose medroxyprogesterone acetate treatment in males with renal or prostatic adenocarcinoma. AB - In 18 patients, 12 with renal and 6 with prostatic carcinoma, the gonadal, pituitary and adrenal functions were studied by measurements of steroid hormones and gonadotrophins, before and after six weeks treatment with medroxyprogesterone acetate (MPA), injected intramuscularly 500 mg per day for 5 days each week. The testosterone-oestradiol-binding globulin (TeBg) was measured and the amount of albumin and TeBg bound and unbound testosterone was calculated. Treatment with high doses of MPA caused a profound decrease in serum concentrations of testosterone, dehydroepiandrosterone sulphate (DHEAS), cortisol and TeBg. There were significant decreases in serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH) and oestradiol-17 beta. The serum concentration of prolactin was significantly elevated. The protein unbound testosterone fraction was lowered by MPA treatment but less than total testosterone. In conclusion, MPA therapy in high dose alters the gonadal, pituitary and adrenal functions suppressing serum concentrations of androgens, gonadotrophins, cortisol and TeBg but elevating prolactin concentration. PMID- 2968649 TI - [Atrial natriuretic polypeptide in the central nervous system]. PMID- 2968650 TI - A camp nurse's perspective. PMID- 2968651 TI - A transcriptional enhancer 3' of C beta 2 in the T cell receptor beta locus. AB - Run-on transcription experiments were used to demonstrate that transcription of T cell receptor beta chain V genes is activated by DNA rearrangement, in a manner similar to immunoglobulin genes. A transcriptional enhancer likely to be involved in this activation has been identified. A 25-kilobase region from J beta 1 to V beta 14 was tested for enhancer activity by transient transfections, and an enhancer was found 7.5 kilobases 3' of C beta 2. The beta enhancer has low activity relative to the simian virus 40 viral enhancer, does not display a preference for V beta promoters, has a T cell-specific activity, and binds two purified immunoglobulin heavy chain enhancer factors. PMID- 2968652 TI - Amyloid beta protein precursor is possibly a heparan sulfate proteoglycan core protein. AB - The amyloid beta protein peptide is a major constituent of amyloid plaque cores in Alzheimer's disease and is apparently derived from a higher molecular weight precursor. It is now shown that the core protein of a heparan sulfate proteoglycan secreted from a nerve cell line (PC12) has an amino acid sequence and a size very similar to those of the amyloid beta protein precursor and that these molecules are antigenically related. This amyloid beta protein precursor related protein is not found in the conditioned medium of a variant cell line (F3 PC12) that does not secrete heparan sulfate proteoglycan. The synaptic localization and metabolism of this class of proteoglycans are consistent with its potential involvement in central nervous system dysfunction. PMID- 2968653 TI - Factor VIII:C purified from plasma via monoclonal antibodies: human studies. PMID- 2968654 TI - [Renal responses to atrial stretch and acute volume expansion in the rat]. PMID- 2968655 TI - [Studies on terminating early pregnancy by laser in rabbits and mice]. PMID- 2968656 TI - [Effect of intracerebroventricular injection of 5-HT on the activity of ATPase and the content of ATP, ADP in the gastric mucosa after stress]. PMID- 2968657 TI - One year of health and social services for adults 40-50 years old with Down's syndrome. A calculation of costs in a delimited area of Denmark. PMID- 2968658 TI - [Cytological screening for the identification of oncological risk groups]. PMID- 2968659 TI - [The means of further improvement of the oncological services in the RSFSR]. PMID- 2968660 TI - [Ultrastructure of atrial myocardiocytes in infectious-allergic myocarditis]. PMID- 2968661 TI - [Interrelations of the clinical departments of a multi-profile hospital]. PMID- 2968662 TI - [Organization of the work of cancer research institutes with regional public health institutions]. PMID- 2968663 TI - [Economic basis of the activities of pulmonological commissions with regard to the detection and treatment of lung cancer]. PMID- 2968664 TI - Degenerative adult onset scoliosis. AB - There are people who have no history of scoliosis who develop spinal deformity of a progressive nature as adults, associated with severe degenerative disc disease. The clinical syndrome associated with this deformity is not well documented. In an attempt to describe this clinical syndrome more precisely, 21 patients with the diagnosis of degenerative scoliosis were identified and reviewed. Review included history with pain drawings when available, physical examination, bone densities, and standing spinal roentgenograms. Patients with spinal compression fractures, spondylolyses, prior history of scoliosis or radiographic findings consistent with an idiopathic scoliosis were excluded. Our review shows that these patients can develop, along with progressive scoliosis, loss of lumbar lordosis with a resulting flat back deformity. These patients commonly present in the sixth decade with a predominantly stenotic symptom complex, but often lack the classic feature of relief in a sitting posture. The number of male and female patients was approximately equal. Roentgenogram findings show a high angle deformity over a short number of spinal segments and an absence of bony features associated with idiopathic scoliosis such as lateral vertebral wedging and alterations of the lamina. The incidence of this condition remains to be established. PMID- 2968665 TI - A digitizing technique for the study of movement of intradiscal dye in response to flexion and extension of the lumbar spine. AB - The effects of flexion and extension exercises on lumbar discs and low-back pain are controversial. Our goals were to develop a technique and program for digitizing and analyzing discograms and to study the motion of intradiscal dye in response to flexion and extension. Thirty-five patients following awake discography were evaluated with lateral radiographs obtained in an extension position and a flexion position. Fifty-three segments with normal morphology and 47 segments with abnormal morphology were studied. Discograms with normal morphology showed numerically significant change in position with a more anterior position occurring during extension. Changes in the position of intradiscal dye in discs with abnormal morphology were less predictable. Digitizing was an advantageous technique. PMID- 2968666 TI - A comparison of CT/discography, pain response and radiographic disc height. AB - CT/discograms of 107 low-back patients were classified by annular degeneration, annular disruption, and pain response. These parameters were compared with the heights of the corresponding discs. Disc height correlated significantly with degenerative annular changes. Comparison of the painless and exact reproduction groups at the L5-S1 level showed a significant increase in exact pain reproduction in narrow discs compared with normal discs. Discs demonstrating slight degenerative changes were often painful but narrowing was detected only when degeneration increased to moderate or severe levels. Some severely degenerated discs were painless and only part of the severe group was narrow. Measuring disc height is a poor method for detecting early, painful degeneration changes. PMID- 2968667 TI - Computed tomography, electrodiagnostic and clinical findings in chronic workers' compensation patients with back and leg pain. AB - One hundred patients with complaints of low-back pain and leg pain, consistent with a diagnosis of sciatica, were evaluated. All patients had complaints for longer than 6 months and had recently undergone electrodiagnostic testing and computed tomography (CT). Correlation was made between symptoms, straight leg raising, clinical neurological deficits, electrodiagnostic and CT findings. The radiation of pain above or below the knee and pain on straight leg raising did not show a high correlation with each other or with neurological deficits or CT findings. Electrodiagnostic studies often defined a radiculopathy in patients with equivocal clinical signs. CT findings did not predict the nature of symptoms or clinical and electrodiagnostic findings. Electrodiagnostic abnormalities showed the greatest ability to predict CT abnormalities. It is concluded that in chronic sciatica patients, no single diagnostic parameter is conclusive and a combination of clinical and laboratory findings is necessary to reach a diagnosis. In addition, many assumptions, valid in patients with acute pain cannot be extrapolated to patients with chronic sciatica. PMID- 2968668 TI - Rehabilitation of the patient with chronic back pain. A search for outcome predictors. AB - This prospective study was initiated 3 years ago to evaluate the outcome and to identify predictors of success or failure in patients admitted to a rehabilitation program for chronic low-back pain. Multiple parameters were evaluated, including psychologic data (MMPI, personal interview, pain drawing, etc.), physical measurements (flexibility, strength and endurance), and demographic data concerning the patient's home and working environment. Information was available on each patient admitted to the program prior to his admission, at completion of the program, 6 weeks following completion of the program and 3 months following completion of the program. A telephone interview was carried out 2 1/2 years following the patient's discharge from the program. Linear regression analysis was used to identify the important independent variables with regard to the dependent variables of relief of back pain, return to work and increased activities at home. Demographic data were of no value as a predictor with the exception of age and returning to work. The patients over the age of 50 returned to work with much less frequency than those less than 50. Psychologic information from the MMPI and similar tests were of no value. The personal preadmission interview of a trained psychologist, however, was a good predictor of an individual's eventual return to work and overall improvement. Worker's Compensation and other litigation was a negative factor in a patient's prognosis. The treatment team's prognosis at the time of discharge from the program was the best overall predictor of a patient's chance of success or failure in the longterm. PMID- 2968669 TI - The use of oral colchicine for low-back pain. A double-blind study. AB - The use of colchicine for the treatment of low-back pain has been controversial; however, recent studies have shown its effectiveness when used intravenously. Studies using oral colchicine alone are lacking. The purpose of this study is to evaluate in a prospective, double-blind fashion the use of oral colchicine in the treatment of low back pain. Group I patients (15) were treated with a placebo capsule, and Group II (12) patients were treated with a colchicine capsule prescribed in a "burst dose" regimen. Patients were evaluated at 1, 2, 4, 6, and 12 weeks with a mean followup of 12 weeks. Parameters studied included patient characteristics, compliance, the McCoy pain drawings, pain analogue scales, the Million scale and objective tests. In terms of therapeutic response, the study shows no statistically significant difference between oral colchicine and placebo. The colchicine group did have an increased number of side effects. PMID- 2968670 TI - [A case report of Behcet disease of incomplete type accompanied by pustulotic arthro-osteitis]. PMID- 2968671 TI - Opioids modulate human neutrophil and lymphocyte function: thermal injury alters plasma beta-endorphin levels. AB - To investigate the role of opioids in the acquired immune dysfunctional state that occurs after burns or trauma, plasma beta-endorphin levels were measured serially in nine severely burned patients, and the effect of four different opioids on normal neutrophil and lymphocyte function was quantitated. The rationale for these studies is that the neuroendocrine system appears capable of interacting with and modulating immune function. The plasma levels of beta endorphin increased to higher than normal during the first 36 hours after burn (15 versus 3.4 pmol/L, p less than 0.05) but quickly returned toward normal. Morphine had the most profound effect on in vitro neutrophil function; it decreased neutrophil chemotaxis but increased neutrophil bactericidal activity for Staphylococcus aureus, as well as resting and zymosan-stimulated oxygen consumption. Other opioids (naloxone, met-enkephalin, and beta-endorphin) had no direct effect on neutrophil chemotaxis or bactericidal activity. Both naloxone and met-enkephalin increased neutrophil oxygen consumption in a dose-dependent fashion, whereas beta-endorphin impaired neutrophil oxygen consumption. None of the opioids altered resting lymphocyte blastogenesis. The only opioid that impaired the ability of normal lymphocytes to respond to mitogen stimulation at physiologically relevant doses was beta-endorphin. These results, documenting that beta-endorphin levels are altered after thermal injury and that opioids can modulate normal neutrophil and lymphocyte function in vitro, support the concept that changes in neuroendocrine activity may occur and potentially alter immune function. PMID- 2968673 TI - [Backache can be removed with back muscle exercises]. PMID- 2968672 TI - Thromboxane receptor blockade in an animal model of ARDS. AB - Manipulation of arachidonic acid metabolism may be important in the prevention and treatment of the adult respiratory distress syndrome. This study evaluated a thromboxane receptor blocker, BM 13.177, in a sheep endotoxin model. Sheep with chronic lung lymph fistulas were pretreated with the blocker before being given endotoxin. The blocker attenuated the early increase in pulmonary artery pressure usually seen after endotoxin (blocker + endotoxin, 31.0 +/- 14.5 mm Hg; endotoxin alone, 42.1 +/- 9.6 mm Hg). There was no effect on lymph flows during the later (permeability) phase (at 6 hours: blocker + endotoxin, 574 +/- 287% baseline; endotoxin alone, 311 +/- 102% baseline). Administration of the blocker alone increased pulmonary artery pressure, arterial pressure, and central venous pressure and decreased heart rate and cardiac index. Thromboxane receptor blockade presumably has little effect on elements of arachidonic acid metabolism other than thromboxane. Thromboxane appears to be important in the early pulmonary hypertension of the sheep endotoxin model but is relatively unimportant in the development of increased pulmonary capillary permeability. PMID- 2968674 TI - [Primary pulmonary hypertension]. PMID- 2968675 TI - [Prospective directions in the organization of pulmonology hospitals]. PMID- 2968676 TI - [Stages in the evolution of bronchopulmonary diseases as the basis for forming groups for dispensary observation and differentiated management of therapeutic, preventive and rehabilitative measures]. PMID- 2968677 TI - [Immediate and late results of rehabilitation at the climatic health resorts on the southern coast of the Crimea with sunlight- and meteorolabile patients with chronic bronchitis]. AB - A study of the social, socioeconomic and economic indices of rehabilitation of patients with chronic obstructive and nonobstructive bronchitis has shown that one should take account of a degree of expression of heliometeolability when selecting patients with chronic bronchitis residing in West Siberia for climatic treatment in the Crimean South Coast. It has been shown that rehabilitation in the Crimean South Coast health resort area is advisable for heliometeostable patients and patients with mild and marked degrees of heliometeolability. Patients with chronic bronchitis should not be sent to the Crimea in winter and late autumn as the rehabilitation effect in this period is lowered. PMID- 2968678 TI - [Effectiveness of immunotropic preparations in chronic bronchitis]. PMID- 2968679 TI - [Organization and results of the treatment of patients with bronchial asthma in Vologda Province]. PMID- 2968680 TI - Do extra-platelet sources contribute to the plasma level of thrombospondin? AB - Thrombospondin, a trimeric glycoprotein contained in the platelet alpha-granules, has been proposed as a marker of in vivo platelet activation. However, it is also synthesised by a range of other cells. The extraplatelet contribution to plasma levels of thrombospondin was therefore estimated by investigating the relationship between plasma thrombospondin levels and platelet count in samples from profoundly thrombocytopenic patients with marrow hypoplasia, using the platelet-specific alpha-granule protein beta-thromboglobulin as control. Serum concentrations of both proteins were highly correlated with platelet count, but while plasma beta-thromboglobulin levels and platelet count also correlated, there was no relationship between the number of platelets and thrombospondin concentrations in plasma. Serial sampling of patients recovering from bone marrow depression indicated that the plasma thrombospondin contributed by platelets is superimposed on a background concentration of at least 50 ng/ml probably derived from a non-platelet source, and plasma thrombospondin levels do not simply reflect platelet release. PMID- 2968681 TI - Effects of N-(1-naphthyl acetyl) piperazine hydrochloride (DQ-2777) on the activation of human plasminogen and the fibrinolysis. PMID- 2968682 TI - [New trends in international invasive cardiology]. PMID- 2968683 TI - [Pelvic instability. A controversial diagnosis]. PMID- 2968684 TI - [Backache after labor. Who will have it and what is the diagnosis?]. PMID- 2968685 TI - [Dealing with current disability pension cases]. PMID- 2968686 TI - High concentration of pancreatic metallothionein in normal mice. AB - An elution profile of supernatant of normal mouse pancreas on a Sephadex G-75 column showed a high peak of zinc in the metallothionein fraction. Cd was added to the supernatant of the mouse pancreas, and 2 cadmium binding proteins were purified by gel filtration and DEAE chromatography. Absorption spectrum and amino acid compositions of the purified proteins agreed with those of MT-I and -II. The concentration of metallothionein in the mouse pancreas was high (227 +/- 11 micrograms/g on 36th day after the birth). The metallothionein concentration in the mouse pancreas was increased by injections of zinc, alloxan or streptozotocin. The results indicate that metallothionein is one of the main zinc binding components in mouse pancreas. PMID- 2968687 TI - [Therapy of rhinophyma]. PMID- 2968688 TI - Pulsed multigated Doppler ultrasonography in the diagnosis of carotid artery disease. AB - To evaluate the accuracy of a pulsed multigated Doppler system, 128 carotid arteries were examined. The spectral broadening index was calculated from the power spectrum of a small sample volume located in the center of the stream according to the flow profile and was related to the degree of stenosis as determined by contrast angiography. Even minor wall irregularities seen on the angiogram were classified as disease. The ability of the system to discriminate between normal and diseased vessels reached a sensitivity of 94% and a specificity of 91%. Classification of greater than 50% or less than 50% stenosis could be performed with a sensitivity of 90% and a specificity of 85%. Pulsed multigated Doppler ultrasonography allows identification of even minor degrees of stenosis of the carotid artery and provides an alternative to duplex scanning. Furthermore, the blood flow profile provided by a multigated Doppler system may add valuable information concerning blood flow characteristics not obtainable by single-gated systems. PMID- 2968690 TI - Transcranial Doppler. AB - TCD recording of flow velocities in intracranial vessels was first described by Aaslid in 1982. The utility of this instrument becomes more apparent as it is used in different clinical settings and compared with angiographic findings (Figures 1 and 2). Its importance in early detection of vasospasm in subarachnoid hemorrhage is now clearly known; increased flow velocity can be documented prior to neurologic deterioration and thus allow early institution of therapy. In patients with stroke or transient ischemic attack of unclear etiology, especially in blacks, Orientals, or females, who have a higher incidence of intracranial arterial disease, TCD can be a very important noninvasive means for detecting stenosis of intracranial vessels. Its value for assessing collateral circulation, intraoperative monitoring, and measuring CBF is quite promising. Hopefully, through further work with TCD, we will be able to clarify the spectrum of its usages as well as its limitations, though the preliminary data indicate that it should be an important addition to present noninvasive evaluations. PMID- 2968689 TI - Use of diastolic velocity ratios to predict significant carotid artery stenosis. AB - Duplex scanning of the carotid bifurcation has emerged as an accurate noninvasive means of predicting and quantifying carotid arterial stenoses. Compared with the more widely reported measurements of spectral broadening and peak frequency ratios, measurements of diastolic velocity ratios have theoretical advantages in predicting carotid artery stenosis. The use of diastolic velocity ratios between the internal and common carotid arteries was prospectively studied in 30 consecutive patients to determine its accuracy in predicting significant stenosis of the internal carotid artery when compared with angiography. A total of 55 carotid bifurcations were studied, and the use of diastolic velocity ratios correctly predicted high-grade stenosis (greater than or equal to 75% diameter reduction) in 52 cases (95%). We conclude that diastolic velocity ratios may be used to accurately detect significant internal carotid artery stenosis. PMID- 2968692 TI - Suppressor cells influence maintenance and reversal of late rejection in indefinitely surviving skin allografts in cyclosporine-treated rats. PMID- 2968693 TI - Impact of a combined regimen of cyclosporine and 3M KCl extracted histocompatibility antigen on heterotopic rat cardiac allograft survival. PMID- 2968691 TI - Alloreactive suppressor T cells and cytokine (gamma-IFN, interleukin 1, 2, and 3) production in cyclosporine-treated mice. PMID- 2968694 TI - The role of suppressor T lymphocytes in the induction of specific immunologic unresponsiveness to rat cardiac allografts by donor leukocytes and cyclosporine. PMID- 2968695 TI - Synergistic immunosuppressive effect of subtherapeutic doses of cyclosporine A on immunologic enhancement in rat recipients of cardiac allografts. PMID- 2968696 TI - Splenectomy antagonizes the action of cyclosporine. PMID- 2968697 TI - Mechanism of prolonged allograft survival induced by ultraviolet B irradiated donor-specific blood transfusions and cyclosporine. PMID- 2968698 TI - Important role of cyclosporine for the induction of immunologic tolerance in adult hosts. PMID- 2968699 TI - Analysis of rejection in cyclosporine-treated cardiac allograft recipients. PMID- 2968700 TI - Cyclosporine and unresponsiveness to allografts induced by polyclonal antilymphocyte serum and donor-specific bone marrow. PMID- 2968702 TI - [Back pain caused by a foreign body]. PMID- 2968701 TI - The effect of cyclosporine on atrial natriuretic peptide in goats. PMID- 2968703 TI - Use of polypropylene mesh for abdominal wall defect in surgery of advanced urachus carcinoma. AB - We report a case of urachus carcinoma in which the resulting abdominal wall defect was reconstructed during surgery with an artificial polypropylene mesh. The use of the mesh made it easier to resect the tumor more radically, even in an advanced case. PMID- 2968704 TI - [Spontaneous bladder fistula of the anterior abdominal wall]. PMID- 2968705 TI - Study of factors mediating effect of photodynamic therapy on bladder in canine bladder model. AB - The canine bladder model was employed to study the factors mediating the effect of photodynamic therapy (PDT) on the bladder. The recovery (time taken for the bladder volume to return to pre-PDT value), gross and microscopic findings, implicate both bladder high filling pressure (60 cm H2O) and high light dose as factors mediating the effect of photodynamic therapy on bladder capacity. We recommend that photodynamic therapy to the bladder be performed under a filling pressure of 30 cm H2O, which is physiologic, and whole bladder illumination at a light dose not greater than 30 J/cm2. PMID- 2968706 TI - [The value of teaching otorhinolaryngology in the training of general practitioners and specialists]. PMID- 2968707 TI - [Regulation of the work of medical personnel of audiology and hearing aid departments (its scientific substantiation)]. PMID- 2968708 TI - [Organizational and methodological aspects of optimal ambulatory ORL surgery]. PMID- 2968709 TI - [Emergency laparoscopy in the examination of middle-aged and elderly patients with acute surgical diseases of the abdominal cavity]. AB - Urgent laparoscopies were performed on 566 elderly and senile patients suspected of acute surgical diseases of abdominal organs. Laparoscopic examinations proved to be highly informative in 90.4%, little informative in 6.6%, erroneous in 3% of cases. Wide use of urgent laparoscopies in elderly and senile patients allowed to avoid laparotomies in 62.3% of the cases. High curative-diagnostic efficiency of laparoscopies added by laparoscopic cholecystostomy and antegrade cholecystocholangiography in patients with acute cholecystitis, cholecystopancreatitis, mechanical jaundice has been shown. PMID- 2968710 TI - [The treatment of thrombophlebitis in a specialized polyclinic]. AB - Under analysis are results of treatment of 150 patients with different forms of acute and chronic thrombophlebitis under conditions of polyclinic. Methods of treatment are described which allowed the time of invalidism to become 2-3 times shorter. PMID- 2968711 TI - Late stages in bacteriophage lambda head morphogenesis: in vitro studies on the action of the bacteriophage lambda D-gene and W-gene products. AB - The in vitro maturation of bacteriophage lambda can be divided into discrete steps. Concatemers of lambda DNA bind terminase to form complex I. This DNA terminase complex then binds a prohead to form a ternary complex (II). Complex II in turn can be converted to infectious phage by the addition of extracts containing the products of the phage genes D, W, FII, as well as phage tails. By using in vitro complementation assays gpD and gpW have been partially purified and their interactions with complex II studied. gpD can bind to complex II in vitro to form a new complex (III) which can be isolated by sedimentation on neutral sucrose gradients. This complex requires only the addition of gpW, gpFII, and phage tails to form mature phage particles. The sedimentation of complex III is virtually identical to that of complex II; however, the resistance of the former to inactivation by DNase is higher, likely due to the partial packaging of the DNA. In similar experiments it was shown that gpW cannot bind to complex II but can effectively interact with complex III. This latter reaction converts complex III to a DNase-resistant form which sediments in a manner identical to that of full phage heads (complex IV). After isolation of the complex IV only gpFII and tails are required for mature phage formation in vitro. gpW is a heat stable protein of molecular weight approximately 10,000. PMID- 2968712 TI - Adenovirus type 12 E1A gene represses accumulation of MHC class I mRNAs at the level of transcription. AB - The levels of the class I antigens and mRNAs of the major histocompatibility complex (MHC) are greatly diminished in cells transformed by adenovirus type 12 (Ad12). Although the Ad12-transforming gene, E1A, is responsible for reduced class I expression, the site at which E1A blocks accumulation of class I transcripts is not known. In this study, we demonstrate by nuclear run-on assays that in Ad12-transformed mouse cells, E1A acts by reducing the rate of transcription of class I genes. PMID- 2968713 TI - Suppression of block to entry into S phase in cell-cycle mutants of rat 3Y1 fibroblasts after transformation by adenovirus type 12. AB - Four temperature-sensitive (ts) mutants of rat 3Y1 fibroblasts, belonging to separate complementation groups, cease to proliferate in the G1 phase of the cell cycle at a restrictive temperature (39.8 degrees). These ts mutants were transformed at a permissive temperature with adenovirus type 12 (Ad12), its E1A gene, or in203S mutant of Ad12 which has a mutation in the E1A 13 S mRNA unique region. We examined whether the proliferation of the transformed cells would be blocked in the G1 phase, at 39.8 degrees. One mutant did not cease to proliferate at 39.8 degrees after transformation with either Ad12 or E1A. In two other mutants, Ad12-transformed cells did not cease to proliferate at 39.8 degrees, whereas E1A-transformed cells did not survive at 39.8 degrees, though they did continue to enter the S phase. Analysis of transcription of the viral early genes in the transformants of one of the latter two mutants suggests that the expression of other viral early genes, in addition to E1A, is required for cell proliferation, in addition to entry into S phase. In the fourth mutant, both Ad12 and E1A-transformed sublines did not cease to enter the S phase but cells readily detached from the dishes. These results suggest that (1) function(s) of the E1A gene alone is sufficient for Ad12 to suppress the inhibition of the initiation of cellular DNA synthesis caused by four different cellular ts defects, (2) functions of Ad12 early genes other than, or in addition to, E1A are necessary for suppression of the inhibition of cell proliferation (and/or for survival) in two of the four ts mutants, and (3) in the case of one other ts mutant, E1A alone overcomes the ts defect and allows for the entire cell proliferation. PMID- 2968715 TI - [Development of specialized medical care in a military district hospital]. PMID- 2968716 TI - [The role of the library of a medical institute in the military patriotic education of students]. PMID- 2968714 TI - Characterization of adenovirus type 40 E1 region. AB - The left-most 3.9 kb of adenovirus type 40 (Ad40) DNA has been sequenced using cloned viral DNA fragments. The Ad40 E1 region is deduced to code for at least four polypeptides, 221 and 249 amino acids as E1A products in addition to 166 and 475 amino acids as E1B products. E1B polypeptides share about 50% homology with well-defined adenovirus types, 2/5, 7, and 12, throughout the E1B sequences. E1A homology of Ad40 to these types is relatively lower than that of E1B, while highly conserved regions of E1A are retained to a certain level in Ad40 as well. Activity for morphological transformation of Ad40 E1A on 3Y1 cells is considerably lower when compared to that of Ad5 and Ad12 E1A genes. Transient chloramphenicol acetyltransferase (CAT) expression assay shows that Ad40 E1A has a trans-acting function, though lower than that of other E1A genes, on adenovirus early promoter. The Ad40 E1A promoter also holds only a little cis-acting activity in 3Y1 cells. Lower activities of both Ad40 E1A promoter and certain E1A functions may explain in part the difficulty in propagation of Ad40. PMID- 2968717 TI - [Blood lipid levels and the nature of the nutrition of 40- to 59-year-old men in the city of Frunze in relation to their ethnic classification]. AB - The relationship between the nutrition character and blood lipid parameters was studied in a male population aged 40-59 years in Frunze. It was established that the levels of total cholesterol, low- and high-density lipoprotein cholesterol in the natives were lower, while those of triglycerides and very low-density lipoprotein cholesterol higher than in the nonnatives. At the same time the correlation values of atherogenic and antiatherogenic lipoproteins in the comparable groups were equally high. In spite of certain differences in the nutrition of the groups investigated (the percent of proteins and complex carbohydrates was higher, while the portion of simple carbohydrates and fat was lower in the ration of the natives than in the nonnatives) actual nutrition of both the native and nonnative populations is characterized by imbalance of certain groups of nutrients in the ration: prevalence of fats and low carbohydrate content. PMID- 2968718 TI - [Enzymatic activity of the microsomal fraction of the mucosa of the small intestine in guinea pigs with vitamin D and C deficiencies]. AB - The influence of vitamin D and C deficiency on the kinetic parameters of sucrase and alkali phosphatase activities was studied in the microsomal fraction of the small intestinal mucosa of guinea pigs. It was found that Km values for these enzymes did not depend on the animal providing with these vitamins. Deficiency of one of these vitamins did not influence sucrase activity, however, simultaneous elimination of vitamins D and C resulted in the activity rise by 92%. Alkali phosphatase and Ca-ATPase activities proved to be similarly dependent on providing with vitamin D in the presence of vitamin C in the ration, while in the absence of vitamin C this dependence was not observed. PMID- 2968719 TI - [Rational organization of nutrition in the rural preschool institutions of Uzbekistan]. PMID- 2968720 TI - [The cloning of regulatory gene mutants of arabinose in E. coli and their DNA sequencing]. PMID- 2968721 TI - Parasitic Isopoda of the Black Sea fishes. PMID- 2968722 TI - Streptokinase therapy for acute myocardial infarction. PMID- 2968723 TI - [Allergic contact dermatitis to prednisolone-21-trimethyl acetate]. AB - A 64-year-old female patient developed periorbital eczema after eye treatment with a corticoid ointment. Epicutaneous tests with the effective component as well as with other ingredients of the ointment and suspected substances revealed a delayed hypersensitivity to prednisolone-21-trimethyl acetate, neomycin, cobalt, and cetyl alcohol. We did not observe any cross-reactions to other corticoids, not even to the closely related prednisolone-21-monoacetate. To the best of our knowledge, allergic contact dermatitis to prednisolone-21-trimethyl acetate has not been described in the literature so far. PMID- 2968724 TI - [Percutaneous transluminal laser angioplasty in peripheral arterial occlusive disease--development of a new laser catheter system]. AB - Percutaneous laser angioplasty in arterial occlusive disease has lately been performed clinically for the first time. Perforation of the arterial wall and formation of aneurysms are serious risks. Two novel laser catheters for laser angioplasty with minimized perforation risk are presented. Catheter I (5F) and II (6.3F) are designed in the same manner. The distal tip of these catheters is ovally formed and marked by a small X-ray dense metal ring. The silica fiber has a core diameter of 400 micron (I) respectively 600 micron (II). Its tip is also marked X-ray densely and therefore the position of the fiber tip can be controlled exactly during laser angioplasty. Using a guide wire and applicating short laser pulses the perforation risk can be minimized. In a total of 132 atherosclerotic stenosed or obstructed human arteries laser angioplasty was performed in vitro using a Nd:YAG laser. There were two perforations (1.5%). The degree of stenosis was reduced from 87 (90)% to 54 (52)%. Using the 600-micron fiber (catheter II) the velocity of laser angioplasty was increased 2.5 times compared to laser angioplasty using the 400-micron-fiber. PMID- 2968725 TI - Selective induction of T-cell subsets by antibodies to the T-cell antigen receptor and to the subset-specific differentiation antigens CD8 and CD4. AB - This report shows that small, resting T lymphocytes from mouse and man can be activated to proliferation and function by submitogenic concentrations of antibodies to T-cell antigen receptor in combination with antibodies to either CD8 or CD4. The combined antibodies can be applied either in solid phase condition or as soluble dimeric heteroconjugates. Activation is more efficient than by high concentrations of anti-T-cell-receptor antibody alone. Most importantly, activation is subset-specific such that the antibody to CD8/CD4 determines which T-cell subset will be induced. PMID- 2968726 TI - [The question of the break of metal-enamel adhesives]. PMID- 2968727 TI - [The presence of Lactobacilli in carious dentin]. PMID- 2968728 TI - [Local enamel fluoridation with hydrogen fluoride or a hydrogen fluoride phosphoric acid mixture]. PMID- 2968730 TI - [Lip pressure measurements in children with cheilognathopalatoschisis]. PMID- 2968729 TI - [Composite-enamel layer systems studied by the ion-beam slope-cutting technic]. PMID- 2968731 TI - [Histopathological study of the effect of a poly(2-hydroxyethyl methacrylate) hydrogel with collagen on calcifying and osteoplastic processes after subcutaneous application in rats]. PMID- 2968732 TI - [The genesis of temporomandibular joint clicking due to pathological changes in the caput mandibulae]. PMID- 2968733 TI - [Wear-resistant plastic teeth. 3. The wear resistance of differently colored plastic teeth]. PMID- 2968735 TI - [The use of vital allogeneic venous transplants for reconstruction of the vestibule of the mouth. An animal experiment study]. PMID- 2968734 TI - [Modification of coronal repositioning plasty for the covering of exposed tooth necks]. PMID- 2968736 TI - [Pelviscopic treatment of tubal pregnancy]. AB - In 1978-1985 we operated on 111 tubal pregnancies pelviscopically. In 12 cases a salpingectomy was performed, in 99 patients the tubes both remained. 17 tubal abortions were extracted, in 82 cases the pregnancy was removed by salpingotomy. One second pelviscopy and one laparotomy were required because of postoperative bleeding. In one patient, a salpingectomy became necessary because of a post operative infection. The pregnancy rate in 45 controlled infertility patients was 53%. A recurrence of ectopic pregnancy in the same or in the other tube occurred in 11%. PMID- 2968737 TI - [Characteristics of the epidemic process manifested as chronic viral hepatitis]. AB - The incidence rates of chronic viral hepatitis in Leningrad over the period of 1962-1984 were studied. The tendency towards a rise in total morbidity because of increased incidence of chronic hepatitis B was shown to appear in recent years. This increase in morbidity was mainly due to its rise among adult males and children, which led to the shift of morbidity to younger age groups. The seasonal rises of morbidity in winter and spring were found to be characteristic of viral hepatitis. PMID- 2968738 TI - [The results of state trials of inactivated influenza vaccines for children]. AB - Controlled epidemiological surveillance covering the total number of 13,355 schoolchildren aged 11-14 years and adolescents was carried out with a view to compare the efficacies of inactivated influenza vaccines. The children were immunized intradermally in a single injection with inactivated influenza vaccines containing A (H3N2) and A (H1N1) hemagglutinins, 3.5 micrograms per 0.2 ml of the preparation each. The studies demonstrated the safety and low reactogenicity of these vaccines, as well as their high antigenic potency. In 1984 during mixed influenza B + A (H1N1) epidemic the preparations produced a pronounced prophylactic effect: the efficacy indices were 1.6-1.9 (p less than 0.001). The results obtained in these studies made it possible to recommend two inactivated influenza vaccines (chromatographic and centrifugal) for practical medicine with the aim of protecting children aged 11 years and over from influenza. PMID- 2968739 TI - [The geometric analysis of the photoballistography of the sagittal movement of the cervical spine]. PMID- 2968740 TI - [The lordosis angle as a predictor of lumbar symptoms. A preliminary study]. PMID- 2968741 TI - [Pathogenetic mechanisms of low backache of disk origin]. PMID- 2968742 TI - [A comparative study of EMG and CAT scan in the lumbo-ischial syndrome. II: Pain in the lumbo-ischial syndrome and the diagnostic value of clinical examination, EMG and CAT scan]. PMID- 2968743 TI - [Osteitis of the pubis]. PMID- 2968744 TI - [Degenerative disk pathology at the level of the cervical spine]. PMID- 2968745 TI - Receptors for insulin-like growth factor-I in plasma membranes isolated from bovine mesenteric arteries. AB - Binding of IGF-I to plasma membranes from bovine mesenteric arteries was studied. The maximal specific binding of IGF-I was found to be 7.4 +/- 1.7% of total 125I IGF-I added to the incubation medium. Unlabelled IGF-I displaced 125I-IGF-I with an IC50 value of 0.5 nmol/l and a maximal displacement of 64.2 +/- 2.8% of total binding. The potency of insulin to displace 125I-IGF-I was 100-1000-fold lower. Cross-linking of 125I-IGF-I to the receptor with disuccinimidyl suberate, followed by SDS-polyacrylamide gel electrophoresis under reducing conditions showed an IGF-I binding protein with a molecular weight of 146,000 Dalton. In summary, we have shown the presence of receptors for IGF-I in plasma membranes isolated from macrovessels. The binding characteristics and the size of the binding unit were found to be similar to those of the IGF-I receptor found in cultured vascular smooth muscle cells. Furthermore, insulin at high concentrations was found to interact with the IGF-I receptor. PMID- 2968746 TI - Differential effects of cisterna magna cannulation on beta-endorphin levels in rat plasma and cerebrospinal fluid. AB - The concentration of beta-endorphin-like immunoreactivity (beta-ELIR) was determined in plasma and cerebrospinal fluid (CSF) of rats at various times following the implantation under Hypnorm anesthesia of a permanent cannula into the cisterna magna. Plasma beta-ELIR levels were highly increased immediately after the operation, and gradually returned to basal within 6 h. The beta-ELIR concentration in CSF followed a completely different pattern. It appeared to be rather stable during the first hour after the operation, was decreased at 2 h, and showed a marked increase 24 h after surgery. Two days after the surgery, beta ELIR levels in CSF had decreased to a stable basal level which was maintained for at least 7 days. When at this stage animals were treated with Hypnorm, plasma beta-ELIR was again highly increased 30 min and not 24 h after injection, but no changes were found in the beta-ELIR concentration in CSF. These data indicate that implantation of a cannula into the cisterna magna induces postoperative changes in the CSF levels of beta-endorphin that are detectable up to at least 24 h later. Moreover, the lack of correlation between CSF and plasma levels of beta ELIR points towards differences in the regulation of brain and pituitary pro opiomelanocortin cells. PMID- 2968747 TI - Effects of plasma volume and osmolality on secretion of atrial natriuretic peptide and vasopressin in man. AB - To clarify the role of blood volume and osmolality in the mediation of the release of atrial natriuretic peptide (ANP) and to examine the relationship between plasma ANP and plasma AVP levels in man, the effects of hypertonic saline and hypertonic mannitol infusion, and of water load on plasma levels of ANP and AVP were studied. Infusion of 5% saline to 7 healthy men at a rate of 0.05 ml.min 1.kg-1 for 2 h resulted in a parallel rise in plasma sodium, osmolality, plasma ANP and plasma AVP, indicating that plasma hyperosmolality stimulates secretion of both ANP and AVP. Infusion of 20% mannitol to 6 healthy men at the same rate resulted in a parallel increase in plasma osmolality, plasma ANP and AVP, whereas plasma sodium decreased, indicating that plasma hyperosmolality stimulates secretion of both ANP and AVP. Water load (20 ml/kg) into 7 healthy men produced a prompt and parallel fall in plasma sodium, plasma osmolality and plasma AVP. In contrast, plasma ANP and plasma volume, calculated from the changes in hematocrit, increased concomitantly, which indicates that expanded plasma volume stimulates secretion of plasma ANP. These results suggest that secretion of ANP in man is regulated principally by plasma volume, which may be modulated by a change in plasma osmolality. AVP secretion, on the other hand, is controlled mainly by osmotic change and secondarily by plasma volume. PMID- 2968748 TI - Atrial natriuretic factor during exercise in patients with congestive heart failure. AB - In order to evaluate the potential relationship between atrial pressure development and release of atrial natriuretic factor (ANF), 33 patients with congestive heart failure were investigated with right-sided heart catheterization during supine graded bicycle exercise. Resting plasma ANF levels were higher in patients with heart failure as compared with normal controls, 75.1 +/- 45.6 pmol/l vs 12.3 +/- 6.2 pmol/l (mean +/- SD, N = 33 and N = 42, respectively) and correlated with right atrial, pulmonary arterial and pulmonary capillary wedge pressures. During exercise, central pressures rose steeply with a simultaneous increase in plasma ANF in all patients. Plasma ANF levels correlated with heart rate at a workload of 25 w, to pulmonary arterial and pulmonary capillary wedge pressure at 50 w, and to pulmonary capillary wedge pressure at 75 w. The increments in ANF levels between the different workloads during exercise did not correlate with the corresponding increments in pressure values. In congestive heart failure, the capability of ANF secretion in consequence to pressure stimuli is preserved, and left atrial pressure seems to be the major stimulus for ANF release during exercise. PMID- 2968749 TI - Corticosteroid-induced stimulation of atrial natriuretic peptide in man. AB - Previously, we reported elevated plasma immunoreactive ANP (irANP) levels from the 2nd to the 9th day of administering either prednisone, 50 mg/day, or 9 alpha fludrocortisone acetate (9 alpha F), 0.6 mg/day, to normal humans. To investigate the course of plasma irANP levels during the first 48 h of corticosteroid administration, 9 healthy men (mean age +/- SEM, 24 +/- 1 years) received in randomised sequence A) a 4-h iv infusion of prednisolone sodium tetrahydrophthalate followed by oral administration of prednisone for 2 days; or B) a 4-h infusion of aldosterone followed by oral administration of 9 alpha F for 2 days. Basal supine plasma irANP levels averaged 32 +/- 5 ng/l in study A and 30 +/- 6 ng/l in study B; they were unchanged or even deceased up to 24 h of glucocorticoid or mineralocorticoid administration, but rose (P less than 0.01) to 56 +/- 9 and 62 +/- 12 ng/l at 48 h, respectively, of the two interventions. During glucocorticoid treatment, blood pressure (BP) and indices of the sodium fluid volume state were unchanged after 48 h. During 9 alpha F administration, body weight increased (1.1 +/- 0.3%, P less than 0.001), whereas urinary sodium excretion (63 +/- 7%, P less than 0.001), hematocrit (4.1 +/- 1.1%, P less than 0.001), and plasma renin activity (38 +/- 4%, P less than 0.001) decreased. CONCLUSIONS: The increase in circulating irANP at 48 h of administration of either a glucocorticoid or a mineralocorticoid demonstrates a distinct but slow response of the ANP system to these corticosteroids in normal humans.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968750 TI - Atrial natriuretic peptide secretion in response to volume expansion and contraction in normal man. AB - The plasma concentrations of ANP in response to isotonic saline-induced volume expansion and furosemide-induced volume contraction in 12 healthy men were measured by radioimmunoassay. The total ANP content was calculated by multiplying the circulating plasma volume (CPV) with the plasma ANP concentration, assuming that most of the ANP remained in the blood vessels. In addition, extracellular fluid volume (ECFV) was measured. After overnight dehydration, plasma ANP concentration significantly increased after isotonic saline was infused, but the urinary ANP concentration did not change essentially. After furosemide-induced volume concentration, plasma ANP concentrations significantly decreased, but the urinary ANP concentrations did not change essentially. As the CVP and ECFV changed, the total plasma ANP also changed. The correlation between these effects was highly significant (r = 0.84 and 0.67, N = 11, respectively). The chromatogram obtained from the plasma in the dehydrated state showed that the area of the high molecular weight ANP peak was relatively large compared with the total area of all ANP peaks. This area of the high molecular weight ANP peaks decreased after saline infusion. In the urine, only the alpha-ANP peak was noted and no high molecular weight ANP peak appeared in the chromatogram. These results indicate that amount and type of ANP secretion may change depending on circulating plasma volume. PMID- 2968751 TI - The bovine placenta: a specific radioreceptor assay for both insulin-like growth factor I and insulin-like growth factor II. AB - Binding of labelled IGF-I and IGF-II was studied to bovine, ovine and human placental cell membranes. The data show a preponderance of type I receptors in human placental membranes, and of type II receptors in ovine placental membranes, confirming reported data. In contrast, bovine placental membranes are rich in both type I and type II receptors. Therefore, the bovine placenta offers a good model for measuring specifically IGF-I (cross-reactivity with IGF-II 7%) and IGF II (cross-reactivity with IGF-I 4%). By Scatchard analysis the apparent Kd (1 1.36 nmol/l) for the high affinity binding sites of the type I receptor is similar in all three preparations. Total binding capacity in ovine placental membranes is, however, 4 times lower. The affinity for the type II receptor is lower than for type I, whereas total binding capacity is higher. Affinity cross linking confirms the competition experiments, showing binding of IGF-I to typical type I and of IGF-II to type II receptors. PMID- 2968752 TI - Plasma atrial natriuretic polypeptide concentration in healthy children from birth to adolescence. AB - We measured plasma atrial natriuretic polypeptide concentrations in the umbilical artery and vein, and peripheral veins of healthy children from birth to adolescence to establish the normal range. The plasma atrial natriuretic polypeptide concentration in the umbilical artery (mean +/- SD, 51.0 +/- 21.4 fmol/ml) was significantly higher than that in the umbilical vein (18.1 +/- 13.5 fmol/ml) in neonates after vaginal delivery. Also neonates aged 5 days or less had a significantly high concentration in the peripheral vein (60.7 +/- 29.4 fmol/ml). There was no significant difference in atrial natriuretic polypeptide concentrations in the peripheral veins between older children and adults. The concentrations in children aged more than 5 days and adults aged 20-34 years were 14.4 +/- 7.4 fmol/ml and 10.0 +/- 4.8 fmol/ml, respectively. However, the atrial natriuretic polypeptide concentration in the umbilical artery was not increased in three neonates delivered by caesarean section although they had a high concentration in the peripheral vein 24 hours after birth. PMID- 2968754 TI - Disability and dependency in elderly people in residential and hospital care. AB - Physical dependency and mental impairment were assessed in 143 residents of residential accommodation for the elderly and 125 patients in geriatric continuing-care wards in South Belfast. Although dependency, immobility and incontinence were more common in the hospital patients, there was considerable overlap in the disability levels in the two types of care. There was evidence of dementia in 51% of the residents and 71% of the patients, while 29% of residents and 70% of patients were incontinent. Overall, less than 1% of the elderly population was in each of residential or geriatric hospital care but the proportion rose with age and was greater in older women. About 17% of elderly people in South Belfast with dementia were in residential or geriatric hospital care. The projected increase in the number of oldest people, particularly women, and the increased risk of dementia in this age group, means that the need for residential and continuing hospital care for the elderly will continue to increase. PMID- 2968753 TI - Acute leukemia of megakaryocyte lineage with tumor formation. An autopsy case of patient with Down's syndrome. AB - In this report we present the autopsy findings of acute megakaryoblastic leukemia with tumor formation in a 2-year-old female infant with Down's syndrome. Chromosomal analysis of blast cells revealed constitutional anomaly of trisomy 21 and two other related types of abnormal clones. Flow cytometric examination revealed blast cells expressing Ia-like or HLA-DR antigens. Postmortem examination showed extensive infiltration of leukemic cells in most of the examined organs, including the bone marrow with myelofibrosis. Tumor masses in the maxillary, frontal and femoral bones and the atria of the heart had undergone massive infiltration of atypical blast cells with an increase in the reticulin network. The final diagnosis was confirmed by ultrastructural cytochemistry of the platelet peroxidase reaction as well as by immunological staining utilizing anti-platelet glycoprotein IIb/IIIa, antiplatelet factor 4 and anti-beta thromboglobulin antibodies for the blast cells. It seems likely that platelet derived growth factor, secondary to an increase in the reticulin network, plays a major role in myelofibrosis of acute megakaryoblastic leukemia with tumor formation. PMID- 2968755 TI - [Measurement of the uveal capillary blood flow by a thermal diffusion method using an electrode with a Peltier stack]. PMID- 2968756 TI - [The bidirectional laser Doppler technic in the measurement of retinal blood flow]. PMID- 2968757 TI - Atrial natriuretic peptide in congestive heart failure. AB - Release of atrial natriuretic peptide (ANP) appears to be a compensatory response in congestive heart failure (CHF) that may counterbalance the adverse effects of stimulated renin-angiotensin and sympathetic nervous systems. We observed increased plasma ANP concentrations in CHF patients in New York Heart Association functional classes II to IV. The fact that such responses already become evident when a patient is in New York Heart Association class II supports the concept that ANP release may counteract the detrimental effects of stimulation of renin and the sympathetic nervous system even in the early phases of heart failure by promoting diuresis and natriuresis, as well as vasodilatation, thus reducing both pre- and afterload. When CHF is severe, however, the counterbalancing effects of ANP may be offset by vasoconstriction and fluid and sodium retention. PMID- 2968758 TI - Three-dimensional network of cords: the main component of basement membranes. AB - Basement membranes were divided into two types: 1) thin basement membranes, such as those of the epidermis, trachea, jejunum, seminiferous tubule, and vas deferens of the rat, the ciliary process of the mouse, and the seminiferous tubule of the monkey, and 2) thick basement membranes, such as the lens capsule of the mouse and Reichert's membrane of the rat. High-magnification electron microscopy was used to examine both types after fixation either in glutaraldehyde followed by postosmication or in potassium permanganate. The basic structure of thin and thick basement membranes was found to be a three-dimensional network of irregular, fuzzy strands referred to as "cords"; the diameter of these cords was variable, but averaged 4 nm in all cases examined. The spaces separating the cords differed, however. In the lamina densa of thin basement membranes, the diameter of these spaces averaged about 14 nm in every case, whereas in the lamina lucida it ranged up to more than 40 nm. Intermediate values were recorded in thick basement membranes. Finally, the third, inconstant layer of thin basement membranes, pars fibroreticularis, was composed of discontinuous elements bound to the lamina densa: i.e., anchoring fibrils, microfibrils, or collagen fibrils. In particular, collagen fibrils were often surrounded by processes continuous with the lamina densa and likewise composed of a typical cord network. Finally, two features were encountered in every basement membrane: 1) a few cords were in continuity with a 1.4- to 3.2-nm thick filament or showed such a filament within them; the filaments became numerous after treatment of the seminiferous tubule basement membrane with the proteolytic enzyme, plasmin, since cords decreased in thickness and could be reduced to a filament, and 2) at the cord surface, it was occasionally possible to see 4.5-nm-wide sets of two parallel lines, referred to as "double tracks." On the basis of evidence that the filaments are type IV collagen molecules and the double tracks are polymerized heparan sulfate proteoglycan, it is proposed that cords are composed of an axial filament of type IV collagen to which are associated glycoprotein components (laminin, entactin, fibronectin) and the double tracks of the proteoglycan. PMID- 2968759 TI - Treatment of locally advanced prostatic carcinoma with LHRH analogues: cytological, DNA-cytophotometrical, and clinical results. AB - From June 1, 1981 to December 31, 1985, 122 patients aged 54 to 83 years, with locally advanced prostatic carcinoma, were treated with buserelin. Nineteen of the patients received combined therapy with buserelin and androcur for the first 3 months. To control the response of the primary tumor to therapy, fine-needle aspiration biopsy of the prostate was made in all patients at 3-month intervals. Fifty-eight (76.3%) of 76 patients with locally advanced prostatic carcinoma, with or without bone metastases, who underwent buserelin therapy for periods of 12-54 months showed good to satisfactory regression grades in the primary tumor. Eighteen patients (23.7%) showed poor regression or none, established by cytological findings and the measure of DNA by means of single cell-scanning cytophotometry. In three of the 58 patients, tumor progression or bone metastases occurred despite favorable regression grade; these were the only cases in which there was a discrepancy between the clinical course of the disease and the grade of regression in the primary tumor. According to TNM classification, 68 of the 78 patients treated for 12-54 months were in stage T3 NX M0; eight were in stage T3/T4 NX M1. On the basis of our long-term studies, it can be stated that buserelin therapy induces positive therapy response in more than 75% of locally advanced, inoperable, primary prostatic carcinoma. The clinical castration caused by buserelin through selective suppression of gonadotrophic secretion in the pituitary gland is, as the term implies, no more effective than surgical castration. However, the gonadotrophin suppression induced by buserelin is reversible and spares the patient the psychic stress of orchiectomy. This is a decisive advantage in light of the fact that in 20-40% of patients with locally advanced primary prostatic carcinoma, the primary tumor is hormone-refractory, and surgical castration would prove unnecessary after all. PMID- 2968760 TI - Results of a Dutch trial with the LHRH agonist buserelin in patients with metastatic prostatic cancer and results of EORTC studies in prostatic cancer. AB - Superactive analogues of luteinizing hormone-releasing hormone (LHRH) have now widely been used in the management of metastatic prostate cancer. In a multicentric study, we have studied the effectiveness of buserelin in patients with previously untreated histologically confirmed metastatic prostate cancer. Fifty-eight patients were enrolled in the study and are now followed for at least 12 months. The treatment consisted of 500 micrograms buserelin administered subcutaneously (s.c.) 3 times daily for the first seven days and thereafter a thrice-daily dose of 400 micrograms buserelin administered intranasally (i.n.). A continuous reduction of testosterone to castrate levels could be obtained in all patients. The following objective responses were seen during the 12 months study period: 5 patients (8.6%) achieved complete regression after 3-10 months (mean 6.4 m); 24 patients (41.4%) achieved partial regression after 1-12 months (mean 4.5 m); 7 patients (12%) remained stable throughout the study; 22 patients (32.3%) developed progression after 2-12 months (mean 7.2 m). Toxicity was minimal. Only one patient dropped out of the study because of side effects. These results are comparable with results of other forms of hormonal treatment of prostate cancer. Long-term results of 3 EORTC GU group comparative studies in metastatic prostate cancer are now available. The first study, 30761, compared cyproterone acetate, medroxy-progesterone acetate, and DES. No differences between the 3 treatment arms were observed except for medroxy-progesterone acetate, which showed less therapeutic effect in the dosage used. Protocol 30762 compared DES and estramustine phosphate. No significant difference in objective response and survival was noted. Protocol 30805 studied a comparison between orchiectomy alone and orchiectomy supplemented by cyproterone acetate and DES. Again no statistically significant difference was found between the three therapeutic arms. PMID- 2968761 TI - Endocrine and clinical evaluation of 107 patients with advanced prostatic carcinoma under long-term pernasal buserelin or intramuscular decapeptyl depot treatment. AB - Three major assumptions emerged from these clinical and endocrine long-term studies. First, buserelin, given pernasally in the conventional doses, and Decapeptyl microcapsules administered intramuscularly in 5-week intervals are equally effective in terms of their long-term castration effect in previously untreated patients with prostatic carcinoma. However, Decapeptyl causes complete LH and subsequent testosterone down-regulation 1 week earlier than buserelin. Furthermore, this treatment is more convenient, and the compliance is better. Both LHRH analogues are equally well tolerated. Second, in groups of prostate cancer patients with far advanced disease treated with palliative intention, only true subjective or objective remission should be considered a positive treatment response. Third, our results comparing PAP and PSA as the two most useful tumor markers with the corresponding testosterone levels suggest a close correlation. PMID- 2968763 TI - Interstitial nephritis in infectious mononucleosis. PMID- 2968762 TI - Stereotaxic biopsy diagnosis of central nervous system lymphoma. AB - Twelve cases of central nervous system lymphoma diagnosed by stereotaxic brain biopsy are reviewed to determine the most effective method to establish the diagnosis given the small amount of tissue obtained by this technique. The stereotaxic biopsy material was examined cytologically, histologically, and immunocytochemically. The diagnosis was established by a smear made at the time of biopsy in eight cases. Histologic sections were diagnostic of lymphoma in 11 of 12 cases. Accurate classification according to the Working Formulation was possible in six cases, with diagnosis of diffuse small non-cleaved non-Burkitt's in three, large cell immunoblastic in two, and mixed small and large cell type in one. Five additional cases were diagnosed as high-grade lymphoma but could not be further subclassified because of the small biopsy size or formalin fixation. Immunocytochemical stains for lymphoid markers on paraffin-embedded material confirmed the diagnosis in ten cases, and, in one of these, the demonstration of monoclonality on air-dried cytospins identified an atypical polymorphous lymphocytic population as neoplastic. PMID- 2968764 TI - Peripheral T-cell lymphoma complicating common variable hypogammaglobulinemia. PMID- 2968765 TI - A 54-year-old woman with fevers, arthralgias, myalgias, and rash. PMID- 2968766 TI - Pathways of Cl- transport in human fibroblasts. AB - Three pathways of Cl- efflux were identified in normal human fibroblasts. Twenty percent of the total Cl- efflux is via an electrically conductive pathway with an efflux constant of 0.016 min-1. This pathway is insensitive to 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and bumetanide but is partially inhibited by anthracene-9-carboxylic acid. Twenty-five percent of the Cl- efflux occurs via Cl- with cation cotransport having an efflux constant of 0.020 min-1. This pathway is inhibited by bumetanide and is dependent on the simultaneous presence of Na+, K+, and Cl-. Under basal conditions, the energetics of this pathway indicate that it is operating close to equilibrium. Fifty percent of the Cl- efflux occurs via an anion exchange pathway having an efflux constant of 0.040 min-1 that is inhibited by DIDS or by removal of Cl- from the extracellular medium. Together these pathways account for 95% of the total Cl- efflux. PMID- 2968767 TI - Effect of short-term DHEA administration on liver metabolism of lean and obese rats. AB - Lean and obese Zucker rats, 7-8 wk of age, were treated with dehydroepiandrosterone (DHEA) for either 3 or 7 days to determine the initial cellular event(s) that might be responsible for the antiobesity activity of DHEA. Epididymal, retroperitoneal, and brown adipose tissue weights were unaltered by either 3 or 7 days of DHEA treatment. Liver weight was not affected by 3 days of treatment but was 13 and 18% higher in 7-day DHEA-treated lean and obese rats, respectively, compared with their corresponding control group. Mitochondrial state 3 respiration rates with glutamate-malate and succinate as substrates were elevated by an average of 35% in 3- and 7-day DHEA-treated obese rats and by 15 20% in 7-day DHEA-treated lean rats compared with rates obtained in the corresponding control groups. State 3 respiration was not affected in 3-day DHEA treated lean rats compared with control lean rats. The specific activities of long-chain fatty acyl-coenzyme A synthase and hydrolase and the levels of free CoA were increased by severalfold in cellular fractions of both DHEA-treated lean and obese rats compared with their respective control group. Hepatic malic enzyme activity, which was shown earlier to be elevated with long-term DHEA treatment, was unaltered by either 3 or 7 days of DHEA administration. The above results suggest the involvement of mitochondrial respiration and fatty acid deacylation/reacylation in the antiobesity mechanism of action of DHEA. PMID- 2968768 TI - Physician acceptance of the hepatitis B vaccine at a university medical center. AB - To examine how physicians at different training levels accepted the hepatitis B vaccine (Heptavax), we mailed questionnaires to all medical students, residents, and faculty at our university medical center. Eighty-one per cent responded; 57 per cent of those had been vaccinated. Attitudes of vaccine safety and efficacy and the perception of increased risk of getting hepatitis B were associated with having been vaccinated. The associations were strongest for the faculty and weak or negative for the medical students. PMID- 2968769 TI - Prevention of exercise-induced asthma by drugs inhaled from metered aerosols. AB - The capacity of salbutamol 0.3 mg (SAL), disodium cromoglycate 3 mg (DSCG), a combination (SAL + DSCG), and ipratropium bromide 80 ug (IB), all given as metered aerosols to prevent exercise-induced (EIA), was compared with that of a placebo, a peroral lactose pill. Seven children participated, having reproducible EIA provoked by running on a treadmill at a heart rate of 170 sustained for 6 min. FEV1 and volume of trapped gas (VTG), defined as the air volume released during rebreathing oxygen with maximum breaths at the end of a multiple breath nitrogen wash out, were used as tests of spasm in large and small airways. SAL and SAL + DSCG offered complete protection in large and small airways. DSCG and IB prevented EIA in large airways (FEV1) to 95%, but only to about 50% in small airways (VTG). SAL or SAL + DSCG gave significantly better protection (FEV1 and VTG) than DSCG and IB (P less than 0.01). Differences between DSCG and IB were not statistically significant. DSCG or propellant caused significant irritation and spasm in small airways (VTG) before exercise. Most subjects seemed to obtain satisfactory protection against EIA by beta 2-agonists. PMID- 2968770 TI - Distribution of myofiber types in the hip and thigh musculature of sheep. AB - The composition of muscles by myofiber type is associated with their locomotory or postural functions. In the present study the composition of the hip and thigh musculature of sheep by myofiber types and the differences in their distribution were examined. Myofibers were classified into type I, IIA, and IIB myofibers by differences in myosin ATPase and NADH tetrazolium reductase (NADH-TR) activity. The vastus intermedius muscle consisted only of type I myofibers, which exhibit weak alkali-stable myosin ATPase and strong NADH-TR activity. The gluteus accessorius and profundus muscles had more than 50% type I myofibers. The other muscles had less than 50% type I myofibers as a whole. Type I myofibers were concentrated in the deep portions of the gluteus and quadriceps femoris muscles, which extend the hip and stifle joints, and of the pectineus muscle. They were scattered evenly in the caudally situated locomotory muscles in the thigh. Type IIA myofibers, characterized by strong alkali-stable myosin ATPase and NADH-TR activity, showed little difference in distribution in the hip and thigh muscles. Type IIB myofibers, characterized by strong alkali-stable myosin ATPase and weak NADH-TR activity, were distributed more in the cranial, caudolateral, and caudomedial portions than in the middle portions of the thigh. The distribution of type IIB myofibers is suited to powerful flexion and extension of the thigh and leg. In the hip and thigh musculature, it appears that type I myofibers are effectively distributed to maintain a standing posture without diminishing the propulsive force of the hindlimb. PMID- 2968771 TI - Plasma concentration of luteinizing hormone, testosterone, dehydroepiandrosterone, androstenedione between birth and one year in the male dog: longitudinal study and hCG stimulation. AB - A longitudinal study from peripheral blood, with samples collected every week, was performed between birth and one year of age on young Fox terriers dogs in order to determine the patterns of plasma LH, T, DHA and A concentrations. T, DHA, A curves show the same profile. The dog model shows the 3 successive steps preceding the adult life that are met in one year time: First, an infantile period between birth and the 12th week of age: the basal level of LH (4.29 ng/ml) and the 3 androgens levels (T less than 0.3 ng/ml, DHA less than 0.45 ng/ml, A less than 0.36 ng/ml) are low. A pubertal period, between the 13th week and the 36th week of age: we observe the maximum activity of the pituitary gland. The basal level of LH (7.97 ng/ml) significantly increase (P less than 0.001). The mean levels of androgens from 17 to 27 weeks of age are still quite low although significantly higher (P less than 0.03) for T and DHA than previously. After the 27th week of age, the androgens concentrations drastically increase. A post pubertal period begins at the 36th week of age. The mean LH (5.85 nh/ml) decrease. The androgens concentrations seem to plateau during the 12th month of age in the range of 2.5-5 ng/ml for T, 1.5-2.5 ng/ml for DHA and 1-2 ng/ml for A. hCG test (35 UI/kg, IM), with samples collected at 6, 12, 24, 30, 36 and 48 hours post injection, were performed at 1, 4, 7, 9 and 12 months of age. At 1 month, only minor variations were noticed; but after 4 months of age, for the 3 steroids, the same time course response was observed as in the adult dog: maximum levels were reached earlier for DHA (6-12 h) than for T and A (24 h). PMID- 2968772 TI - Hemodynamic effects of primary closure of omphalocele/gastroschisis in human newborns. AB - To determine whether they could establish reliable, objective criteria that would predict safe, primary closure of abdominal wall defects (omphalocele/gastroschisis) in newborn infants, the authors measured intraoperative changes in intra-gastric pressure (IGP), central venous pressure (CVP), cardiac index (CI), systolic arterial blood pressure (BP), and heart rate (HR). Eleven neonates, who averaged 2.7 kg (range 1.5-4.1 kg) and 36 weeks gestation (range 30-41 weeks) were anesthetized with fentanyl (7.5-12.5 micrograms/kg), metocurine (0.3 mg/kg), and oxygen. Three infants had defects that were too large to close primarily. Of the eight infants who underwent primary closure, four required re-operation within 24 h because of oliguria or poor peripheral perfusion. Infants who required re-operation had intra-gastric pressures of 20 mmHg or more, a decrease in CI of 0.78 1.min.m2 or more, and an increase in CVP of 4 mmHg or more. Heart rate, BP, and systemic vascular resistance did not differ in infants requiring and not requiring re-operation. The authors conclude that intraoperative measurement of changes in IGP, CVP, and/or CI can reliably predict success or failure of primary operative repair of abdominal wall defects in human neonates. PMID- 2968773 TI - Flow recovery and vasoconstriction following microvascular anastomosis. AB - A variety of factors may limit blood flow recovery in free flaps. In this study one of these factors, vasoconstriction, and its consequences was investigated. Blood flow recovery at the skin level following microvascular anastomosis was evaluated in the rat orthotopic groin free flap model using a laser Doppler velocimeter. In a study of 41 rats, measurements of external vessel diameter were made using a standard machinist's drum micrometer. After each flap was raised and the anastomoses performed, the field was flooded with either saline, 2% lidocaine, or 0.75% bupivacaine (Marcaine). The vascular clamps were released and measurements of flow were recorded as a percentage of the control value. In the saline-control group, it was found that flow gradually recovered to 91% of the preoperative value in an average time of 5.7 minutes. Six of 12 vessels in this group were found to have measurable spasm of the pedicle. In the lidocaine treated group, flow recovered to 93% of the preoperative value in an average of 5.0 minutes. Two of 15 vessels had measurable spasm. In the Marcaine-treated group, flow recovered to 94% of the preoperative value in an average of 5.1 minutes. Three of 14 vessels showed measurable spasm. The final level of flow recovery showed no statistical difference in any of these groups whether or not spasm was present. We therefore conclude that, in this model, alteration in the diameter of the vascular pedicle as a consequence of performing a microvascular anastomosis affects only the time it takes to achieve ultimate recovery of flow but does not affect its final level. PMID- 2968774 TI - Combined plastic and gynecological surgical procedures. AB - The purpose of the present study was to determine the extent to which the planned combination of one of several major plastic and gynecological surgical procedures may alter patient morbidity and postoperative hospitalization when compared with similar procedures carried out separately. A review of 10 consecutive years of operative procedures produced 63 patients who underwent combined gynecological and plastic surgical procedures. Thirty-three of the combined-surgery patients could be suitably matched with 2 control patients each to compare complications and length of hospital stay. The requirement for transfusion was significantly greater (p = 0.01) in the combined-surgery versus the paired-control groups. No other complications, however, were increased in the combined-surgery patients. There was a significant reduction (p = 0.005) in hospital stay of 1.92 postoperative days for two groups of combined-surgery patients versus the control groups. In all combined-surgery patients, patient response was favorable. Recommendations on preoperative planning and intraoperative and postoperative management are discussed. There is a perception in the surgical community that combining procedures unacceptably increases morbidity. For the past 12 years plastic and gynecological surgeons in our institution have offered combined procedures to our patients, with positive results and favorable patient response. A search of recent medical literature demonstrated several references to the planned combination of plastic and gynecological or general surgical procedures [2, 4, 5]. However, the only study with matched control patients dealt exclusively with abdominoplasty in association with one of five major gynecological procedures. PMID- 2968775 TI - Hypersensitivity dermatitis following suction-assisted lipectomy: a complication of local anesthetic. AB - We report a case of severe dermatitis involving the abdomen and thighs following suction-assisted lipectomy of these areas wherein local anesthetic containing the preservative methylparaben was used for infiltrative anesthesia. This use of local anesthetics with epinephrine can be of value in the performance of suction assisted lipectomy to reduce blood loss, serve as an adjunct to other intraoperative anesthetic techniques, and for postoperative analgesia. Local anesthetic solutions commonly contain additives, which serve as antioxidants and antimicrobials. The most common of these preservatives is methylparaben, which can cause delayed hypersensitivity reactions. These reactions may be neither recognized nor clinically significant in small areas of infection, whereas in large body surface infiltrative procedures, such as suction-assisted lipectomy, these reactions may be of considerable consequence. This article reviews the pathophysiology and treatment of these reactions and gives recommendations for avoiding them. PMID- 2968776 TI - Monoclonal antibody analysis of mononuclear cells in myopathies. V: Identification and quantitation of T8+ cytotoxic and T8+ suppressor cells. AB - In polymyositis (PM) and inclusion body myositis (IBM), but not in dermatomyositis there is evidence of cell-mediated cytotoxicity: T8+ cells accompanied by macrophages focally surround, invade, and destroy nonnecrotic muscle fibers. However, the T8 marker appears on both cytotoxic (Tc) and suppressor (Ts) cells. The Leu-15 marker appears on Ts but not on Tc cells, but it also appears on macrophages and on some killer/natural killer cells. To obviate this problem, the T8, Leu-15, and Leu-7 markers were demonstrated by sequential paired immunofluorescence in single cryostat sections. Using this approach, we reliably differentiated for the first time between Tc and Ts cells in tissue sections. Six cell phenotypes were identified: T8+ Leu-15-Leu7- Tc cells, T8+ Leu-15+ Leu-7- Ts cells, three types of Leu-7+ killer/natural killer cells, and T8-Leu-15+ Leu-7- macrophages. Muscle specimens from 5 patients with PM and 5 with IBM were studied. In each case, 6 nonnecrotic muscle fibers focally surrounded and invaded by mononuclear cells were selected randomly. A total of 2,022 mononuclear cells were analyzed, 870 from patients with PM and 1,152 from those with IBM. When counts of the identified cell phenotypes in individual patients were pooled, there were four times as many T8+ Leu-15- Leu-7- Tc cells as T8+ Leu-15+ Leu-7- Ts cells in either PM or IBM samples. However, when the relative frequencies of the Tc and Ts cells were examined in individual patients, the Tc cells tended to become more abundant, and the Ts cells correspondingly less abundant, with the duration of symptoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968777 TI - Mismatched dsRNA (ampligen) induces protection against genomic variants of the human immunodeficiency virus type 1 (HIV-1) in a multiplicity of target cells. AB - Mismatched double-stranded RNA of the form r(I)n.r(C12-U)n (Ampligen) has been shown to be active against human immunodeficiency virus type 1 (HIV-1) using CEM and C3 cells as targets for infection by the highly similar HIV-1 isolates HTLV IIIB and LAV (Montefiori, D.C. and Mitchell, W.M., 1987, Proc. Natl. Acad. Sci. U.S.A., 84, 2985-2989). The scope of Ampligen's anti-HIV-1 activity was examined in this study using the genetically divergent HIV-1 isolate HTLV-IIIRF, two additional target T-cell lines, H9 and MT-2, and a monocyte/macrophage cell line, U937. As judged by indirect immunofluorescence, reverse transcriptase activity and vital dye uptake, Ampligen was active against HTLV-IIIRF in H9, MT-2, C3 and U937 cells in addition to being active against HTLV-IIIB in U937 cells. A minimum of 1 h preincubation of cells (MT-2) with Ampligen was required for maximum activity. These results suggest that Ampligen's potential clinical efficacy may not be limited by either the highly variable nature or host cell range of HIV-1. PMID- 2968778 TI - [Targeting chemotherapy of colorectal cancer by monoclonal antibody conjugated antitumor drug]. AB - Neocarzinostatin (NCS) was conjugated with mouse monoclonal antibody (A7) that specifically reacted with human colorectal carcinomas in vivo and the conjugates (A7-NCS) were administered to patients with colorectal carcinoma. In several cases the size of the metastatic liver tumor was decreased after intraarterial infusion of A7-NCS. No severe adverse effects were observed in any patients who received A7-NCS. These results suggest that missile cancer chemotherapy using monoclonal antibody as a carrier of anticancer drug will be a promising method for use against cancer. PMID- 2968779 TI - [Targeting chemotherapy with transferrin-neocarzinostatin conjugate]. AB - In efforts to obtain preferential uptake of anticancer agents into tumors, we explored the possibility of delivering such agents by transferrin receptor mediated endocytosis. Human diferric transferrin was conjugated with neocarzinostatin (NCS) using N-succinimidyl 3-(2-pyridyldithio)-propionate. This conjugate is capable of binding to the transferrin receptor and is internalized by endocytosis. The inhibitory effect of the conjugate on cell growth was remarkable when a human colorectal cancer cell line, M7609 was used as a target in vitro. In addition, in vivo efficacy of the conjugate in inhibiting the growth of M7609 cells implanted subcutaneously into nude mice was also observed when the conjugate was administered through a tail vein. The observed toxicity was a transient decrease in the red blood cell count, which returned to normal within 14 days. The half disappearance time of the conjugate was 55 min, while that of free NCS was 7 min. No serious side effects with regard to liver or kidney function were detected. This conjugate is an appropriate model for the receptor mediated delivery of ligand-drug complex and may be useful for future clinical application. PMID- 2968780 TI - [Progestogen therapy in the treatment of endometrial cancer--clinical results and mechanism of steroid action]. AB - The use of medroxyprogesterone acetate (MPA) in therapy of patients with endometrial cancer has been recently examined by the Japan Gynecological Cancer Treatment Group. The response rate of oral MPA was 23.6% (13/55 evaluables) and the average period up to the onset of response was 15.2 weeks (4-28 wks). The response rate was higher in well-differentiated tumors than in poorly differentiated ones. Although it is well known that steroid hormone action is mediated by steroid hormone receptor, the presence of progesterone receptor in cancer tissue seems to be not absolutely necessary for the responsibility to MPA, a potent progestational compound. The reason was studied, in vitro, in our laboratory by using the technique of human tumor clonogenic assay. In this system, MPA showed a marked suppressive effect on the colony formation of endometrial cancer cells. Similar effects were also observed with progesterone, 17 alpha-hydroxyprogesterone caproate and megestrol acetate. Norethindrone and norgestrel, which are both potent progestational compounds, showed almost no anticancer activity on Ishikawa cell in this in vitro system. It became clear that progestational activity was not always associated with anticancer activity, and norethindrone had no influence on the inhibitory effect of MPA, although norethindrone has a strong affinity for progesterone receptor. Similarly, RU 486, which is known as a potent antiprogestogen agent and has a high affinity to progesterone receptor, did not influence the effect of MPA. These results clearly indicated that the anticancer activity of MPA was not mediated by high affinity low capacity progesterone receptor, and a pharmacological effect must be considered for understanding the effect of MPA on endometrial cancer. PMID- 2968781 TI - Basal cell carcinoma mimicking rhinophyma. Case report and literature review. AB - An 82-year-old man presented with rapid enlargement of a long-standing rhinophyma. Following an uncomplicated excision of rhinophyma, histologic examination showed that the enlargement was entirely due to basal cell carcinoma. Radiotherapy was administered, and the nose has now healed satisfactorily. There are few reports of an association between rhinophyma and basal cell carcinoma; but it is claimed that it occurs more frequently than is expected by chance. The evidence for this is reviewed. Previous reports were anecdotal or contained biases that prevent generalization of the results and, in addition, a statistical analysis was incorrect. There is insufficient evidence to claim an association between basal cell carcinoma and rhinophyma. PMID- 2968782 TI - Immunohistology of rheumatoid nodules and rheumatoid synovium. AB - The immunohistological features of rheumatoid nodules and rheumatoid synovium were examined using monoclonal and polyclonal antibodies raised against macrophages, HLA-DR, leucocyte common antigen, and immunoglobulin components. The palisading cells surrounding the necrotic centre of the rheumatoid nodule were shown to be HLA-DR positive leucocytes, mostly histiocytes. The inflammatory infiltrate associated with rheumatoid nodules showed many immunohistochemical similarities to that of rheumatoid synovium, including a preponderance of IgG positive plasma cells, and a similar number and microanatomical pattern of distribution of HLA-DR positive cells. The significance of these findings for the cellular immunopathology and aetiology of the rheumatoid lesion is discussed. PMID- 2968783 TI - Syndrome of diffuse abnormal insertional activity: case report and family study. AB - In 1979 Wiechers and Johnson described ten patients with diffuse abnormal insertional activity on EMG examination in the absence of neuromuscular disease. We present a family group with identical findings. The propositus is a 53-year old woman who presented with back pain. EMG studies revealed trains of positive sharp waves with needle movement in all muscles studied. Nerve conduction studies, radiographs, and laboratory studies were all unremarkable. We recruited eight additional family members who underwent a screening EMG of five muscles. Four patients had trains of positive sharp waves present in all five muscles. To our knowledge, this is the first report confirming the findings of Wiechers and Johnson. We concur with them that the abnormality appears to be genetically transmitted in an autosomal dominant pattern. Although without clinical significance, it is important for electromyographers to be aware of this entity so as not to mistakenly ascribe serious neuromuscular disease to these patients. PMID- 2968784 TI - Presymptomatic neuropsychological impairment in Huntington's disease. AB - Ten asymptomatic individuals at risk for Huntington's disease (HD) were determined by the use of linked DNA probes to have a high (HD+ group) or low (HD- group) probability of having inherited the mutant gene. Neuropsychological examination, performed without knowledge of DNA results, revealed impairments in five of seven subjects in the HD+ group. Abnormalities were related to visuospatial abilities or to functions associated with the frontal lobes. All three subjects in the HD- group showed no neuropsychological impairment. Statistical analyses confirmed differences between the HD+ and HD- groups. Affected parents of subjects were at least 12 years older at symptom onset. These results demonstrate that clear neuropsychological impairment may be present in HD even when overt signs and symptoms are not expected for a number of years. PMID- 2968785 TI - Doppler flow measurements in the assessment of carotid artery stenosis. PMID- 2968786 TI - Fibrinogenolysis in the absence of fibrin formation in severe hypobaric hypoxia. AB - Acute exposure to hypoxia causes acceleration of activated partial thromboplastin time (aPTT) and a rise in factor VIII precoagulant activity (F VIII:C). To determine whether this activation of coagulation leads to in vivo fibrin formation we investigated 15 army pilots before and at the end of 21 min (range 14-29) of hypobaric hypoxia. Mean final pressure in the decompression chamber was 283 (250-310) mm Hg causing a fall in oxygen saturation to 61.5% (53-69). Hypobaric hypoxia caused acceleration of thrombin time (p less than 0.05), aPTT (p less than 0.01), and euglobulin lysis time (p = 0.05), as well as a rise of F VIII:C (p less than 0.05), beta-thromboglobulin (p less than 0.005), fibrin(ogen) degradation products E (p less than 0.005) and B beta 15-42 (p less than 0.001), as well as lactate (p less than 0.001). Fibrinopeptide A, a marker of in vivo fibrin formation, did not change significantly. It is concluded that severe hypoxemia due to rapid decompression going to the limit of tolerance does not lead to fibrin formation, whereas the rise in fibrin(ogen) degradation products demonstrates activation of the fibrinolytic system. PMID- 2968787 TI - Contribution of blood and systemic circulation to the processing of pro-(atrial natriuretic factor). AB - Atrial natriuretic factor-(Asn1-Tyr126)-peptide, the 13.6 kDa propeptide of atrial natriuretic factor (ANF), is stored in the secretory granules of atrial cardiocytes. ANF-(Ser99-Tyr126)-peptide, the 28-amino-acid species, is the circulating form of this hormone in the rat. As the site of maturation of the prohormone is still unknown, the present study was undertaken to understand the contribution of the circulation to the maturation process of pro-ANF. 125I-ANF (Asn1-Tyr126)-peptide was incubated with whole rat blood, plasma or serum for different time intervals, and the products were analysed. There was minimal activation of the propeptide in either whole blood or plasma. Incubation with serum, however, resulted in the formation of an 11 kDa and a 3 kDa peptide which corresponded respectively to the N-terminal and C-terminal parts of the propeptide. These results suggest that hydrolysis of the propeptide in serum is brought about by enzymes that may be stimulated during coagulation but which may not play a major role in the activation of pro-ANF in the circulation. Plasma analysis at different time intervals after prohormone injection indicated a non specific hydrolysis of the pro-ANF molecule. The disappearance rate curves, obtained with radiolabelled pro-ANF, suggested the presence of two components with half-lives of 2.1 +/- 0.4 min and 52.5 +/- 8.4 min respectively. A metabolic clearance rate of 1.49 +/- 0.22 ml/min and an initial distribution volume of 47.4 +/- 8 ml were calculated. These results indicate that the maturation of pro-ANF to its active circulating form takes place before it is released into the circulation. PMID- 2968788 TI - Hydrolysis of histones by proteinases. AB - Hydrolysis of histones by proteinases from rat liver, skin and other sources was studied by using a rat thymus histone preparation as the substrate and polyacrylamide-gel electrophoresis and densitometric analysis as the methods to detect histone subtypes and their hydrolysis. The rat mast-cell proteinase I effectively hydrolysed histones except type H4. Thrombin hydrolysed effectively histones H1 and H2A, whereas plasmin hydrolysed all types of histones. Cathepsin D hydrolysed especially histone H2A. Cathepsins B and L hydrolysed all histones more slowly, and cathepsin H hydrolysed them extremely slowly. Epidermal aminoendopeptidase did not hydrolyse histones. Trypsin and chymotrypsin were used as reference enzymes, which hydrolysed all types of histones in very low concentrations. This study suggests that a variety of proteinases could play a role in histone hydrolysis. Hydrolysis of a specific subtype of histones, such as histone H2A at pH 6 by cathepsin D, may be directly involved in regulation of epidermal-cell differentiation. PMID- 2968789 TI - Synthesis of lysine-containing sulphonium salts and their properties as proteinase inhibitors. AB - Some sulphonium salts derived from lysine were synthesized with the general structure R-Lys-CH2S+-(alkyl)2. They were examined as inhibitors of the cysteine proteinase clostripain, which has a preference for cleaving peptide bonds at the carboxy group of basic amino acids, and of a number of trypsin-related serine proteinases. Clostripain was irreversibly inactivated by all reagents examined, but in the case of the serine proteinases, depending on the reagent structure, irreversible and reversible inhibitions were observed. These were kinetically characterized. PMID- 2968790 TI - 67 k calcimedin (67 kDa) is distinct from p67 calelectrin and lymphocyte 68 kDa Ca2+-binding protein. AB - The 67 k calcimedin is a Ca2+-binding protein present in both muscle cells and peritoneal macrophages. Many tissues, including lymphoid tissues, liver and lymphocytes, have been shown to contain Ca2+-binding proteins of similar molecular size, such as the p67(67 kDa) calelectrin or the 68 kDa lymphocyte protein. We have tested affinity-purified antibodies raised to the smooth-muscle 67 k calcimedin in these several tissues and here report that the 67 k calcimedin is not detectable in liver, thymus, spleen or thymic lymphocytes. These findings support recent biochemical evidence, discussed here, suggesting that the 67 k calcimedin is a protein different from calelectrin and the 68 kDa lymphocyte protein. The more limited tissue distribution of the 67 k calcimedin, which includes muscle and macrophages, suggests that the 67 k calcimedin may function in Ca2+-mediated events special to these cell types. The affinity-purified antibodies to the 67 k calcimedin will be useful in obtaining information concerning the special roles of this Ca2+-binding protein in these cells. PMID- 2968792 TI - The effect of atrial natriuretic factor on natriuresis in acutely hypophysectomized saline-loaded rats. AB - Acute hypophysectomy (a.h.) (i.e. pituitary ablation 120 min prior to the experiment) abolished the ability of anaesthetized rats to excrete sodium following isotonic saline load (4% body weight). The diuretic response remained even slightly increased as compared to control sham-operated animals (s.o.) (maximal UNaV a.h. rats +/- SE: 0.65 +/- 0.10 mumol/min vs maximal UNaV in s.o. rats: 4.54 +/- 0.50 mumol/min). Alpha h-atrial natriuretic factor 1-28 (ANF) in a doses of 3.3 micrograms/animal.40 min restored natriuresis in a.h. rats following isotonic saline load to the level observed in s.o. rats (4.25 +/- 0.64 mumol/min), the change being transient. A decrease of blood pressure to 78.6 +/- 3.21 mm Hg during ANF infusion was followed by a reduction in the natriuretic effect of ANF, while the diuretic effect remained unchanged. Based on own results and literary data it could be suggested that impairment of ANF release or its renal effect are involved in the abolishment of homeostatically effective natriuresis in a.h. animals. PMID- 2968793 TI - Control of sodium excretion: atrial natriuresis and mineralocorticoid escape: some new aspects. PMID- 2968791 TI - Wheat-germ-agglutinin and Ricinus communis-agglutinin-binding sites of BHK cells compared with each other and with 140 kDa fibronectin receptors. AB - We compared the wheat-germ agglutinin (WGA) and Ricinus communis agglutinin (RCA) binding sites of baby-hamster kidney (BHK) cells. There were 1.01 X 10(8) WGA binding sites per cell (Kd = 0.027 nM) and 6 X 10(6) RCA-binding sites per cell (Kd = 0.014 nM). Binding of WGA or RCA to BHK cells resulted in more than 75% of the cell-surface binding sites becoming associated with the cytoskeleton (i.e. resistant to extraction with detergent), although no more than 10% of these sites were associated with the cytoskeleton before addition of the lectins. After binding of WGA to the cells, the cell surface was cross-linked so extensively that it remained intact even after detergent extraction of the treated cells, and could be observed by electron microscopy. A similar cross-linking effect did not occur after binding of RCA to cells, which may be because there were so many more binding sites for WGA than for RCA. The composition of WGA- and RCA-binding molecules was analysed by lectin affinity chromatography of metabolically radiolabelled BHK cells. We found that in the WGA-binding-molecule preparations there were eight major polypeptides, ranging in molecular mass from 93 to 340 kDa, and that the RCA-binding molecules were a subpopulation of the WGA-binding molecules. A polyclonal antibody against the 140 kDa fibronectin (FN) receptors of Chinese-hamster ovary (CHO) cells immunoblotted a 145 kDa polypeptide component in both WGA- and RCA-binding-molecule preparations. The results indicated that the 145 kDa component was present in at least two FN-receptor complexes that differed in glycosylation, only one of which was able to bind to RCA affinity columns. The oligomeric nature of the FN-receptor complex, which contained three polypeptides with molecular masses of 120-145 kDa, was demonstrated by using anti-(CHO-cell FN receptor) antibodies to immunoprecipitate extracts prepared from radioiodinated BHK cells. PMID- 2968794 TI - Free and bound forms of atrial natriuretic peptide (ANP) in rat plasma: preferential increase of free ANP in spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP). AB - Free and bound forms of atrial natriuretic peptide (ANP) in rat plasma were analysed by gel permeation chromatography combined with a radioimmunoassay (RIA) for rat ANP (rANP). Gel permeation chromatography showed two immunoreactive peaks in rat plasma, one corresponding to alpha-rANP, rANP(99-126), and the other eluted at a high molecular weight, clearly different from gamma-rANP, rANP(1 126). The chromatographic profile of rat plasma after incubation with synthetic alpha-rANP demonstrated that the high molecular immunoreactivity had ANP-binding capacity. This bound form of ANP was almost totally excluded following extraction procedure, therefore, the immunoreactive ANP (ir-ANP) measured with the extraction assay was mainly free ANP. On the other hand, direct RIA may detect not only the free but also the bound form of ANP. Using both direct RIA and the extraction method, bound forms of plasma ANP in spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP) were compared to normotensive Wistar Kyoto rats (WKY). Bound forms of plasma ANP in 20-week-old SHR and SHRSP were significantly higher than that in age-matched WKY. The ratio of free/bound form of plasma ANP in SHR and SHRSP also significantly increased compared to WKY, indicating a preferential increase in free ANP in the plasma of these hypertensive rats. These findings suggest that a bound form of ANP may be present in rat plasma and that it may play some pathophysiological role in the hypertension of SHR and SHRSP. Increased free ANP in plasma may indicate a compensatory increase in ANP release in these hypertensive rats. PMID- 2968795 TI - Inhibition of phospholipase A2 activity of guinea-pig alveolar macrophages by lipocortin-like proteins purified from mice lung. AB - Guinea-pig alveolar macrophages were harvested by bronchoalveolar lavage and purified by differential adhesion. They were labeled with 14C-Arachidonic acid and then exposed to platelet-activating factor or to the calcium ionophore A23187. The activity of cellular phospholipase A2 was considered as the release of free 14C-Arachidonic acid in the cell supernatant. The pretreatment of guinea pig alveolar macrophages with two lipocortin-like proteins (36 kDa and 40 kDa) purified from mice lung induced a significant inhibition of their phospholipase A2 activity upon platelet-activating factor and calcium ionophore stimulation. These results indicate that lipocortin-like proteins can modulate the phospholipase A2 activity of isolated cells in vitro. PMID- 2968796 TI - Rapid purification of two lipocortin-like proteins from mice lung. AB - We have purified two proteins (40 kDa and 36 kDa) from mice lung by the method of calcium-precipitation/EGTA solubilization and then a separation on a high anion exchanger column (Mono Q HR 5/5. Pharmacia) with a gradient of NaCl. The two proteins were strong inhibitors of phospholipase A2 as assessed in vitro with porcine pancreatic phospholipase A2 and [3H]-oleic acid labeled E. Coli membranes as substrate. The 40 kDa protein had a pI of 5.8 and was found to be immunologically related to human recombinant lipocortin I. The 36 kDa protein had a pI of 4.7 and cross-reacted with a polyclonal antibody raised against a 32 kDa human lipocortin-like protein described in human blood mononuclear cells. We report here a rapid purification of two distinct lipocortin-like proteins from mice lung. PMID- 2968797 TI - Inactivation of the mitochondrial adenosine triphosphatase from Trypanosoma cruzi by oxygen radicals: role of thiol groups. AB - Inactivation of Trypanosoma cruzi mitochondrial ATPase by oxygen radicals, generated by redox cycling of the ascorbate-Cu system (Cataldi de Flombaum, M.A. and Stoppani, A.O.M. (1986) Biochem. Int. 12, 785-793), involves oxidation of the enzyme thiols, as indicated by the competitive kinetics obtained with p chloromercuribenzoate, a selective SH-reagent. Dithiothreitol prevented the ascorbate-Cu effect but did not reactivate the enzyme. Non-competitive kinetics were obtained with ascorbate-Cu and increasing MgATP concentration, or with phenylglyoxal, as second inhibitor. Since phenylglyoxal reacts with arginyl residues at the ATPase hydrolytic site, these results suggest that the oxgen sensitive thiols were located outside the hydrolytic site. PMID- 2968798 TI - The increase in viscosity and peroxidation of sarcoplasmic reticulum membrane lipids in isadrine myocarditis. AB - The viscosity of membranes isolated from sarcoplasmic reticulum of rabbits with isadrine myocarditis was studied, using pyrene as a hydrophobic fluorescent probe. The increase in the viscosity of membranes from injured heart occurred at lower temperatures and was sharper than in the case of intact heart in both "free" and "bound" lipid domains. The increase in the lipid viscosity under myocarditis was associated with decreased Ca++, Mg++ -ATPase and cAMP-dependent protein kinase activities and with an elevated content of lipid peroxidation products. PMID- 2968799 TI - Effects of phospholipases C on membrane-bound enzymes of yeast. AB - Alkaline phosphatase was released from protoplasts of the yeast Saccharomyces cerevisiae without cell lysis not only by phosphatidylinositol (PI)-specific phospholipase C but also by phosphatidylcholine (PC)-hydrolyzing phospholipase C. Activities of mitochondrial enzymes such as succinate dehydrogenase, antimycin sensitive NADH-cytochrome c reductase, and oligomycin-sensitive ATPase were decreased by the action of PC-hydrolyzing phospholipase C. Hydrolysis of microsomal PC or PI did not cause any decrease in the activities of NADPH cytochrome c reductase and antimycin-insensitive NADPH-cytochrome c reductase. In the requirement of phospholipids, the properties of yeast mitochondrial enzymes were very close to those of mammalian mitochondrial enzymes, whereas those of yeast microsomal enzymes were completely different from those of mammalian microsomal enzymes. PMID- 2968800 TI - Inhibition by suramin of mitochondrial ATP synthesis. AB - Suramin, a drug intensively used in the chemotherapy of African trypanosomiasis and onchocerciasis, is currently being tested in clinical trials for AIDS treatment. Its effects on mitochondrial energy metabolism in mammals were studied. At low concentrations it inhibited ATP synthesis and ATPase activity in submitochondrial particles, as well as ADP-stimulated oxygen consumption and the uncoupler-stimulated ATPase activity in intact rat liver mitochondria. At higher concentrations it also inhibited uncoupled electron transport in both submitochondrial particles and intact mitochondria. From comparison of the kinetic patterns of those inhibitions, evidence suggesting that the adenine nucleotide translocase may be another target for the action of suramin was obtained. The relevance of these findings to the understanding of the biochemical basis of suramin toxicity is discussed. PMID- 2968802 TI - [IgE and anaphylaxis mediators]. PMID- 2968801 TI - Calcium channel blocker inhibition of the calmodulin-dependent effects of thyroid hormone and milrinone on rabbit myocardial membrane Ca2+-ATPase activity. AB - The Ca2+-ATPase activity of rabbit myocardial membranes is stimulated in vitro by L-thyroxine and by milrinone, a bipyridine. These effects are concentration dependent and calmodulin requiring. The calcium channel blockers nifedipine and verapamil have been reported to have anti-calmodulin effects in other assay systems. In this study we have examined the effects of nifedipine and verapamil on rabbit myocardial membrane Ca2+-ATPase activity, in the absence (basal activity) and presence of exogenous L-thyroxine (T4), 10(-10) M, and milrinone, 10(-7) M. Basal enzyme activity was inhibited by a minimum of 10(-6) M nifedipine (IC50 of 3.4 X 10(-5) M) and 10(-5) M verapamil (IC50 of 1.5 X 10(-4) M). Both calcium antagonists inhibited enzyme stimulation by T4 and milrinone, with half maximal inhibition of T4 and milrinone effects, respectively, at 2.9 X 10(-5) M and 9.0 X 10(-6) M nifedipine and 3.0 X 10(-5) M and 5.2 X 10(-5) M verapamil. The addition of exogenous purified calmodulin, 40 ng/micrograms membrane protein, in the presence of 10(-5) M nifedipine or verapamil restored T4-stimulated enzyme activity. Nifedipine and verapamil, each at a concentration of 10(-6) M, significantly inhibited binding of radioiodinated calmodulin to rabbit heart membranes in vitro. These studies provide evidence that nifedipine and verapamil have an anti-calmodulin effect in this myocardial enzyme system. Through interaction with calmodulin, the channel blockers inhibit thyroid hormone and milrinone stimulation of myocardial membrane Ca2+-ATPase. PMID- 2968803 TI - [Inflammatory cells and hypersensitivity reactions]. PMID- 2968804 TI - Importance of plasma renin activity suppression and venous distensibility on pressor and natriuretic responses to intravenous salt load in borderline hypertension. AB - Cardiopulmonary receptors influence renin release in a variety of physiological situations and in a fashion related to the degree of peripheral venous distensibility. We studied two groups of borderline hypertensives (BHTs) with different capacities to suppress plasma renin activity in response to saline infusion (0.20 mL/kg/per minute for 2 hours). Those BHTs with low suppressive capacity (L-supp) showed an increased venous distensibility in comparison with those with high suppressive capacity (H-supp). Saline infusion led to a significant increase in blood pressure only in L-supp BHTs, which was associated with enhanced 24-hour postloading natriuresis and raised plasma levels of an Na/K ATPase inhibitor (+12.2%). This result underlines the importance of venous distensibility as a determinant of pressor and humoral response to acute volume expansion. PMID- 2968805 TI - Changes in the renin-angiotensin system, exchangeable body sodium, and plasma and atrial content of atrial natriuretic factor during evolution of chronic renal failure in the rat. AB - The remnant kidney model of progressive renal failure in the rat was used to assess the relationships between the renin-angiotensin system, exchangeable body sodium, and both plasma concentration and atrial content of atrial natriuretic factor (ANF) measured sequentially over 4 weeks. Following subtotal nephrectomy plasma creatinine (mumol/L) rose from 40 +/- 6 (sham) to 107 +/- 24 (P less than 0.05) at 1 week, and rose further to 124 +/- 20 (P less than 0.05) by 4 weeks. Plasma renin activity (nanograms of angiotensin I/mL/min) rose from 4.5 +/- 0.5 to 11.8 +/- 2.8 (P less than 0.05) at 1 week, but was suppressed by 4 weeks to 2.2 +/- 0.3 (P less than 0.001). Plasma angiotensin II (pg/mL) was 52 +/- 2 (sham), 117 +/- 20 at 1 week (P less than 0.05) and 51.3 at 4 weeks. Exchangeable sodium (mmol/kg) rose progressively from 43.2 +/- 5 (before surgery) to 48.6 +/- 0.9 at 1 week and 50.8 +/- 2.2 at 4 weeks. Plasma atrial natriuretic factor (ANF) (pg/mL) rose progressively from 114 +/- 6 (sham) to 248 +/- 31 (P less than 0.01) at 1 week and 456 +/- 78 at 4 weeks (P less than 0.01). Atrial ANF content fell as the plasma concentration rose. In the remnant kidney model of progressive renal failure there was a progressive increase in exchangeable body sodium and plasma atrial natriuretic factor, with reciprocal changes in atrial ANF content suggesting that ANF secretion rate was increased. Initially the renin-angiotensin system was stimulated, but later it was suppressed. PMID- 2968806 TI - Effect of cyclosporin A on 2ry MLR in patients with progressive systemic sclerosis. AB - We have studied the effects of the immunosuppressive agent Cyclosporin A (CsA) on the allogenic stimulation of human lymphocytes from 10 patients with progressive systemic sclerosis (PSS) and 10 healthy controls, when activated in vitro during 1ry and 2ry MLR. CsA markedly suppressed 1ry MLR in both PSS subjects and controls. In contrast to the marked suppression of 2ry MLR observed for most of the PSS subjects and all control subjects, lymphocytes from 3 out of 10 PSS patients had paradoxically amplified allogenic responses when primed in the presence of CsA. The possibility is discussed that in these patients the recruitment of suppressor-effector cells by CsA-resistant suppressor-inducer lymphocytes is impaired. PMID- 2968807 TI - Suppression of immune-mediated otitis media by T-suppressor cells. AB - Effects of mucosa-derived T-suppressor cells on induction of IgG-mediated otitis media with effusion (OME) were investigated in mice, since antigen-antibody reactions in the tympanic cavity are pathogenic mechanisms of OME. Splenic T suppressor cells from strain C3H/HeN female mice, to which ovalbumin (OVA) had been orally administered, were transferred intravenously to syngeneic mice. The mice receiving the T-suppressor cells were immunized with OVA intraperitoneally and then injected with OVA into the tympanic cavity. Nine of ten control mice, to which splenic T cells from saline-fed mice were administered, developed OME, while OME was seen in only one of ten mice receiving splenic T cells from OVA-fed mice. The results show that IgG-mediated OME can be suppressed to a certain extent by the induction of antigen-specific, mucosa-derived, T-suppressor cells. PMID- 2968808 TI - Positron emission tomography imaging of dopamine and serotonin receptors in Huntington's chorea. PMID- 2968810 TI - [Sleep apnea in severely handicapped children: a clinical and electrophysiological study]. PMID- 2968809 TI - The diagnosis of dioxin-associated illness. AB - The clinical recognition of dioxin-associated illness can be extremely difficult for the physician. After analyzing the relative sensitivity and specificity of reported manifestations of exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), we suggest criteria for the diagnosis of dioxin toxicity. Exposure to higher doses of 2,3,7,8-TCDD may lead to the appearance of chloracne and the increased excretion of porphyrins and porphyria cutanea tarda. Liver function abnormalities, peripheral neuropathy, hyperlipidemia, and evidence of weakness and depression may occur following exposure; however, these findings are less specific since diseases such as diabetes or alcoholism could cause several of these problems. The long-term effects of exposure to low-dose TCDD are currently uncertain. PMID- 2968811 TI - Distribution of the CD45R antigen in the maturation of lymphoid and myeloid series: the CD45R negative phenotype is a constant finding in T CD4 positive lymphoproliferative disorders. AB - We have compared the expression of the common leucocyte antigen (CD45) and the restricted leucocyte antigen (CD45R) on normal haematopoietic cells, cell lines, and a total number of 136 cases of myeloid and lymphoid proliferative syndromes. CD45, the conventional leucocyte antigen, presents a generalized distribution along the lymphoid and myeloid maturation pathway with the exception of some myelomas and pre-B leukaemias. In contrast, the expression of the CD45R determinant is more limited. Although it is found in the majority of the differentiation stages of B cells and monocytes, it is present only in the early stages of myeloid differentiation. On T cells it is expressed on mature thymocytes and in the majority of CD8+ lymphocytes and a subset of CD4+ cells on peripheral blood. Finally, our results also indicated that CD4+ T lymphoproliferative syndromes are derived from the CD4+ CD45R- subset (20/20 cases). PMID- 2968812 TI - Decreased expression of the common acute lymphoblastic leukaemia antigen (CALLA/CD10) on neutrophils from patients with thermal injury. AB - The common acute lymphoblastic leukaemia antigen (CALLA/CD10) is a normal component of the circulating neutrophil cell surface membrane. In order to examine the potential functional significance of CALLA/CD10 we analysed the expression of this molecule on neutrophils isolated from thermal injury patients, since these patients have a well-documented constellation of neutrophil defects affecting their microbicidal functions. Expression of neutrophil CALLA/CD10 was monitored by indirect immunofluorescence and flow cytometry. We observed that CALLA/CD10 expression was quantitatively reduced on burn patient neutrophils, compared to healthy donors (P less than 0.001). In contrast, burn patient neutrophils expressed normal levels of class I HLA molecules and the C3bi receptor. Reduced expression of CALLA/CD10 was not associated with neutrophil activation or exposure to plasma 'factor(s)' in vivo. Analysis of normal bone marrow neutrophils by cell sorting indicated that expression of CALLA/CD10 occurs late in neutrophil maturation, since 25% of polymorphonucleated bone marrow neutrophils did not express cell surface CALLA/CD10. Attempts to examine the chemotactic responses of CALLA/CD10 positive and negative neutrophils from burn patients were hampered by previous exposure of these cells to chemoattractants in vivo. Collectively, our findings suggest that burn patient peripheral blood neutrophils may be deficient in CALLA/CD10 due to insufficient maturation time in the bone marrow following thermal injury. PMID- 2968813 TI - Functional properties of an isolated alpha beta heterodimeric human placenta insulin-like growth factor 1 receptor complex. AB - Treatment of human placenta membranes at pH 8.5 in the presence of 2.0 mM dithiothreitol (DTT) for 5 min, followed by the simultaneous removal of the DTT and pH adjustment to pH 7.6, resulted in the formation of a functional alpha beta heterodimeric insulin-like growth factor 1 (IGF-1) receptor complex from the native alpha 2 beta 2 heterotetrameric disulfide-linked state. The membrane-bound alpha beta heterodimeric complex displayed similar curvilinear 125I-IGF-1 equilibrium binding compared to the alpha 2 beta 2 heterotetrameric complex. Triton X-100 solubilization of the alkaline pH and DTT-pretreated placenta membranes, followed by Bio-Gel A-1.5m gel filtration chromatography, was found to effectively separate the alpha 2 beta 2 heterotetrameric and alpha beta heterodimeric IGF-1 receptor species, 125I-IGF-1 binding to both the isolated alpha 2 beta 2 heterotetrameric and alpha beta heterodimeric complexes demonstrated a marked straightening of the Scatchard plots, compared to the placenta membrane-bound IGF-1 receptors, with a 2-fold increase in the high affinity binding component. Similar to the membrane-bound IGF-1 receptor species, the 125I-IGF-1 binding properties between the alpha 2 beta 2 heterotetrameric and alpha beta heterodimeric complexes were not significantly different. IGF-1 stimulation of IGF-1 receptor autophosphorylation indicated that the ligand dependent activation of alpha beta heterodimeric protein kinase activity occurred concomitant with the reassociation into a covalent alpha 2 beta 2 heterotetrameric state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968814 TI - Binding of glycogen, oligosaccharides, and glucose to glycogen debranching enzyme. AB - The binding of glucose and a series of oligosaccharides to glycogen debranching enzyme was determined by the ability of the saccharides to decrease the rate of reaction of sulfhydryl groups with 5,5'-dithiobis(2-nitrobenzoate) (DTNB). At pH 7.2, the strength of binding increases with chain length from glucose to maltotriose to maltopentaose but not to maltohexaose, and the free energies for binding of the oligosaccharides suggest subsites of equivalent affinities for the four glucose units following the initial reducing moiety. The rate of reaction of DTNB with enzyme saturated with saccharide is the same for all compounds, suggesting that all the saccharides, including glucose, induce the same conformational state. The site of binding may be that which binds the alpha-1,6 linked side chain of the natural limit dextrin substrate. At pH 8.0, this site exhibits similar characteristics, but an additional site, which may bind the four terminal glucose units of the main chain of the natural substrate, is manifested and exhibits different characteristics, including a very low affinity for glucose itself. The binding of glycogen to the debranching enzyme was monitored by centrifugal separation from the protein and exhibits a much lower dissociation constant than that for the oligomers, suggesting that branched polymers have more than one set of subsites. PMID- 2968815 TI - Acid-stable trypsin-plasmin inhibitors formed enzymatically from plasma precursor protein. AB - Enzymatic formation of acid-stable trypsin-plasmin inhibitors (ASTPIs) in human plasma with several proteinases, particularly SH-proteinases, was demonstrated. The maximal activity obtained with bromelain was 40 U/ml plasma, which corresponded to about a 10-fold increase as compared to the untreated control plasma (4.2 U/ml). Gel filtration revealed at least two ASTPI activity peaks of molecular weight 16,000 (main peak) and 8000 (minor peak). The main ASTPI was further purified by trypsin-Sepharose affinity chromatography, isoelectric focusing and gel filtration on Sephadex G-75 superfine. The purified inhibitor was found to be identical to the active fragment of plasma ASTPI or urinary trypsin inhibitor (UTI) formed by bromelain treatment. It had an isoelectric point (pI) of 3.7, a molecular weight of 16,000 by SDS-polyacrylamide gel electrophoresis and was a glycine- and glutamic acid-rich protein lacking histidine. The NH2-terminal amino acid sequence was H2N-(Lys)-Glu-Asp-Ser-X-Gln Leu-Gly-Tyr-Ser-Ala-Gly-Pro-X-Met-Gly-Met-Th r-X-Arg - Tyr-Phe-Tyr-... COOH, which was homologous to the Lys22-Met36 part (or Glu23-Met36 part; 30% of the total) of the plasma ASTPI or UTI molecule (molecular weight 70,000-80,000 by gel filtration). The purified ASTPI displayed the same antigenicity as UTI and exerted strong inhibitory effects on trypsin, chymotrypsin and plasmin amidolysis, but had a much lesser effect on plasmin fibrinolysis. It also strongly inhibited non-plasmic fibrinolysis with human leukocyte proteinase and earthworm proteinase. PMID- 2968817 TI - Regulation of axonemal Mg2+-ATPase from Paramecium cilia: effects of Ca2+ and cyclic nucleotides. AB - Ciliary activity is regulated by Ca2+ and cyclic nucleotides, but the molecular mechanisms of the regulation are unknown. We have tested the ability of Ca2+ and cyclic nucleotides to alter ciliary Mg2+-ATPase or to stimulate phosphorylation of axonemal dynein. Mg2+-ATPase activity in cilia and axonemes from Paramecium was stimulated 2-fold by micromolar Ca2+, but this Ca2+ sensitivity was lost upon solubilization of the dyneins from the axoneme. The Ca2+-sensitive component of ciliary Mg2+-ATPase activity was inhibited by the dynein inhibitors vanadate and Zn2+, but was insensitive to the calmodulin antagonists calmidazolium and melittin. Dynein activity in the high-salt extract from axonemes was also insensitive to calmidazolium. Calmodulin did not sediment with 22 S or 12 S dyneins on sucrose gradients containing Ca2+, but it did sediment in the region from 19 S to 14 S. Mg2+-ATPase activity in ciliary fractions was unaltered in the presence of cAMP or cGMP. However, polypeptides associated with the 22 S and 12 S dyneins, as well as proteins of 19 S, 15 S, and 8 S, were substrates for endogenous ciliary kinases. High molecular weight polypeptides that sedimented at 22 S and 19 S were phosphorylated in a cyclic nucleotide-stimulated manner. PMID- 2968816 TI - Purification and properties of dyneins from Paramecium cilia. AB - Dynein ATPases were purified from Paramecium cilia by salt extraction followed by sucrose density gradient centrifugation and anion exchange chromatography. The two major dyneins sedimented in sucrose gradients as species of 22 S and 12 S. After purification by anion exchange chromatography, their specific activities were about 0.4 and 0.5 mumol/min per mg, respectively. The dyneins could be distinguished by subunit composition and immunological crossreactivity. Sucrose density gradient centrifugation revealed additional ATPase activity in the region between the 22 S and 12 S dyneins, including a 19 S activity. Mg2+-ATPase activities of the dyneins and the 19 S activity were inhibited by vanadate and Zn2+, and were activated by Triton X-100. Antibodies against the 22 S dynein from Paramecium reacted on immunoblots with most of the polypeptides of 22 S dynein, and showed that the heavy chains of 22 S dynein are not identical to those that sediment at 19 S and 12 S. Several minor ATPase activities were revealed by anion exchange chromatography of fractions from the 22 S, 19 S and 12 S regions of sucrose gradients. These minor activities were stimulated by Mg2+, inhibited by vanadate, and could be distinguished from each other by their elution positions and polypeptide compositions. PMID- 2968818 TI - The Doppler flow velocity waveform in the fetal internal carotid artery with respect to fetal behavioural states. A longitudinal study. AB - Doppler flow velocity waveforms in the fetal internal carotid artery were recorded in 21 normal pregnancies during fetal behavioural state IF (quiet sleep) and 2F (active sleep) according to Nijhuis and co-workers, from the 36th week of gestation onwards. The present study confirms the earlier finding of a significant reduction of the pulsatility index (PI) in state 2F as compared to state 1F at 37-38 weeks of gestation and demonstrates that this difference in PI exists throughout the entire period, in which fetal behavioural states have been described. Furthermore, it is demonstrated that there is a significant reduction in PI of this vessel during the last 4 weeks of gestation, suggesting a haemodynamic redistribution, favouring blood supply to the brain during the latter weeks of gestation. PMID- 2968819 TI - [Suppressors of the graft vs graft reaction in tolerance to alloantigens]. AB - Immune response and suppressor cell activity of CBA (H-2k) mice made tolerant to allogeneic C57B1/6 (H-2b) heart graft were studied in graft-versus-graft reaction (GvGR). Intact CBA spleen cells inhibited response of (CBA X C57B1/6)F1 cells to antigenic stimulus (sheep red blood cells--SRBC), when injected together into lethally irradiated (CBA X C57B1/6)F1 mice. Spleen cells of tolerant mice were unable to decrease immune response of (CBA X C57B1/6F1 lymphocytes to SRBC and suppressed specifically the inhibition induced by intact CBA spleen cells. Spleen cells from tolerant mice were also capable of suppressing GvGR induced by CBA lymphocytes immune to C57B1/6 cells. Pretreatment of tolerant spleen cells with rabbit antithymocyte globulin and complement before adoptive transfer diminished markedly the suppression. The results obtained in the study suggest that suppression of transplantation immunity in this model is mostly due to T suppressor cells. PMID- 2968820 TI - [Humoral mechanisms of in vitro inhibition of mouse lymphocyte immunoreactivity by mastocytoma P815 cells]. AB - The inhibitory capacity of mastocytoma cell line P815 and its cultural supernatant (CS) was studied in the reaction of blast transformation (RBT) and mixed lymphocyte culture (MLC). An addition of both P815 cells and CS resulted in dose-dependent inhibition of lymphocyte proliferation in RBT and MLC. The treatment of DBA/2 spleen cells with CS for 2 h at 37 degrees C resulted in a significant decrease in proliferative activity and induction of supressor cells. PMID- 2968822 TI - Cytogenetic findings and clinical features in acute leukemia and transient myeloproliferative disorder in Down's syndrome. AB - Cytogenetic, immunologic, and electron microscopic studies were performed on the blast cells of 28 pediatric patients with Down's syndrome, 13 with acute leukemia (DS-AL) and 15 with transient myeloproliferative disorders (DS-TMD). Clonal chromosome abnormalities were found in the cells of all patients with DS-AL but not those with DS-TMD. The younger ages and higher hemoglobin concentrations, platelet counts, and WBC counts of DS-TMD patients provided a clinical contrast with the frankly leukemic cases. Myelodysplastic syndrome, characterized by a small percentage of leukemic blast cells, was observed in 11 of the 13 patients with DS-AL compared with none in the DS-TMD group. Electron microscopy disclosed a positive platelet peroxidase reaction in each of the 11 DS-TMD patients and in nine of the 13 DS-AL patients. Immunologic studies revealed antiplatelet megakaryocyte antigens on the blast cells of the majority of patients in both study groups. Our findings suggest that the blast cells in cases of DS-AL and DS TMD arise from cells of the megakaryocytic lineage or from a myeloid progenitor with the capacity for megakaryocytic differentiation. The high risk of the development of AL in patients with DS who are less than 3 years old may be related to increased megakaryocyte proliferation in this age group. PMID- 2968821 TI - [Analysis of the composition of skeletal muscle fibers in skaters' muscles]. AB - Muscle fibers composition was investigated in vastus lateralis muscles of 103 skaters. In the total population of skaters muscle fibers composition was as follows: muscle fibers, type I--in 54 +/- 2%, muscle fibers, type II A--in 35 +/- 1% and muscle fibers, type II B--in 11 +/- 1%. 13 +/- 8% muscle fibers, type I; 1.56 +/- 6% muscle fibers, type II A and 31 +/- 7% muscle fibers, type II B were observed in the muscles of sprinters, while the muscles of long-distance skaters contained 60 +/- 4%, 33 +/- 4% and 7 +/- 2% muscle fibers, respectively. PMID- 2968823 TI - Decrease of natural killer cell activity and monokine production in peripheral blood of patients treated with recombinant tumor necrosis factor. AB - Tumor necrosis factor (TNF), a protein predominantly produced by activated macrophages/monocytes, is presently available in recombinant, purified form for clinical trials. Intensive studies in many laboratories have shown that besides the tumorcytotoxic effects, TNF acts on a large array of different cells and has potent immunomodulatory activities. In a clinical phase I study, some immunologic functional parameters of blood cells from patients who received 24-hour infusions of recombinant human TNF (rhTNF) were analyzed. Natural killer (NK) cell activity, TNF production, interleukin-1 (IL-1) production and mitogen-induced proliferation were measured either in whole blood samples or in cultures of peripheral mononuclear leukocytes of the patients directly before and after rhTNF infusion. NK cell activity, TNF and IL-1 production capacity and proliferative responses to concanavalin A (Con A) were significantly reduced after rhTNF application. We conclude from these observations that rhTNF in vivo acts directly or indirectly on NK cells and monocytes by either inactivating their functional capacity or by absorbing the relevant cells to the endothelial cell layer, thus removing them from circulation. PMID- 2968824 TI - Glycoprotein IIb-IIIa and RGD(S) are not important for fibronectin-dependent platelet adhesion under flow conditions. AB - Previous studies have indicated that activated blood platelets interact with fibronectin through binding of fibronectin to the glycoprotein IIb-IIIa complex (GPIIb-IIIa). The cell attachment site of fibronectin with its crucial arg-gly asp(-ser) [RGD(S)]sequence is involved in these bindings. We studied the importance of these interactions for the fibronectin dependence of platelet adhesion under flow conditions. An RGDS-containing hexapeptide (GRGDSP) was compared with a nonreactive control peptide (GRGESP). The GRGDSP-peptide inhibited thrombin-induced aggregation and adhesion under static conditions at 0.1 mmol/L. This concentration had no effect on platelet adhesion to nonfibrillar collagen type I in flow. GRGDSP at 1 mmol/L had a significant inhibitory effect at 1,500 s-1, but not at the lower shear rates of 800 and 300 s-1 where platelet adhesion is also fibronectin dependent. On the matrix of cultured human umbilical vein endothelial cells, 1 mmol/L GRGDSP had no effect on platelet adhesion. The relation between GPIIb-IIIa and fibronectin dependence was investigated with platelets of a patient with Glanzmann's thrombasthenia and monoclonal antibodies to GPIIb-IIIa using endothelial cell matrix (ECM) as a surface. Platelets of normal controls or a patient with Glanzmann's thrombasthenia showed a similar inhibition of adhesion in the presence of fibronectin-free plasma after the ECMs had been preincubated with antifibronectin F(ab')2 fragments. Incubation of platelets with anti-GPIIb-IIIa showed inhibition of platelet adhesion at high shear rates. Dependence on fibronectin for platelet adhesion was still observed even though separate experiments had shown that these anti-GPIIb-IIIa antibodies could block binding of radiolabeled fibronectin to thrombin-activated platelets. These data suggest the existence of another binding system for the interaction of platelets with fibronectin that may only appear when fibronectin is present on a surface. PMID- 2968826 TI - The acceptability of cochlear implants and vibrotactile aids. AB - To obtain data on the likely acceptability of cochlear implants (and of vibrotactile aids), a questionnaire was sent to profoundly/totally hearing impaired patients. The patients were asked how they felt about their current communication problems and possible benefits to be obtained from such devices. Of the 153 replies obtained, the combined 'yes' or 'maybe' responses indicated that 58% would accept an implant. The corresponding figure for vibrotactile aids was 73%. The statistically significant factors correlating with cochlear implant acceptance were the patient's expectation of the benefit it would give, the time needed for rehabilitation and the degree of communication difficulty that they suffer. The only significant factor in acceptance of a vibrotactile aid was whether the patient had attended the IHR clinic. This implies that the underlying factor is the patient's knowledge about these devices. PMID- 2968825 TI - In vivo platelet activity and serum albumin concentration in nephrotic syndrome. AB - To clarify the relationship between serum albumin concentration and in vivo platelet activity in nephrotic syndrome, Beta-thromboglobulin (B-TG) levels and circulating platelet aggregation ratio (PAR) were determined in 25 nephrotic patients. PAR levels were significantly decreased compared with the controls and showed a positive correlation with serum albumin concentrations. The values of B TG were high in all nephrotic patients and showed an inverse relationship with serum albumin levels. In addition, increased B-TG levels correlated with decreased PAR values. Therefore serum albumin plays a regulatory role in the activity of circulating platelets in the nephrotic syndrome. Prospective longitudinal studies must be done to elucidate the usefulness of platelet inhibitors as a prophylactic therapy of intravascular thrombosis in nephrotic patients. PMID- 2968827 TI - Dehydroepiandrosterone and its sulfate enhance memory retention in mice. AB - Dehydroepiandrosterone (DHEA) and its sulfate (DHEAS), major naturally occurring precursors of both androgenic and estrogenic steroids, were shown in the present study to have convincing memory enhancing effects in mice. Post-training intracerebroventricular (i.c.v.) administration of DHEA in dimethylsulfoxide (2 microliters) prevented the amnesia for footshock active avoidance training (FAAT) caused by the same volume of dimethylsulfoxide alone. DHEAS significantly enhanced retention of FAAT in weakly trained mice whether injected i.c.v. or s.c. immediately post-training or given in the drinking water for a 2-week period. In the latter instance DHEAS was shown to facilitate retention of FAAT without enhancing acquisition. The maximally effective doses were: i.c.v., 162 ng/mouse; s.c., 700 micrograms/mouse; and oral, 1.45 mg/mouse/day. DHEAS administered i.c.v. occluded the amnestic effects of anisomycin (inhibitor of protein synthesis) and scopolamine (muscarinic cholinergic antagonist). There was a time dependence of the facilitatory effects of post-training i.c.v. administration of DHEAS on retention of FAAT, significant enhancement of retention being observed when it was given either immediately (within 2 min) or at 30 and 60 min after training, but not at 90 or 120 min. DHEAS given i.c.v. also improved retention for step-down passive avoidance. In all instances, dose-dependent inverted U curves were obtained in a manner typical for memory enhancing substances. At a practical level, these experiments open new possibilities for the development of substances that may help in alleviating amnesic disorders in man. PMID- 2968828 TI - Dopamine D1 receptors in the nucleus accumbens: involvement in the reinforcing effect of tegmental stimulation. AB - Subtypes of dopamine receptors involved in the mechanism of reinforcement were examined. Twenty-five rats were implanted with bipolar electrodes into the ventral tegmental area (VTA) and cannulae into the nucleus accumbens (NAc) or the caudate-putamen complex (CPu). The rats were trained to press a bar to receive VTA stimulation, and dopamine antagonists were injected into the brain while the animals were responding. SCH 23390 (D1-specific antagonist) injected into NAc ipsilateral to the electrode site suppressed responding, but injections into contralateral NAc or ipsilateral CPu had no effect. Haloperidol was less effective. Sulpiride (D2-specific antagonist) produced no effect when injected into NAc, though slightly suppressed responding when injected into CPu. The animals that have reduced their response rate after accumbens injection of SCH 23390 responded at a normal rate when the frequency of pulses in the tegmental stimulation was increased, indicating that they were not incapacitated by the D1 antagonist. These findings suggest that dopamine D1 receptors in the nucleus accumbens are critically involved in the reinforcement produced by ventral tegmental stimulation. PMID- 2968830 TI - Dentistry, the law and AIDS. PMID- 2968829 TI - Dehydroepiandrosterone sulfate improves memory in aging mice. AB - Middle-aged (18 month old) and old (24 month old) mice showed poorer retention of footshock active avoidance training (FAAT) than young mice (2 month old). Immediate post-training subcutaneous injection of dehydroepiandrosterone sulfate (DHEAS) improved retention of FAAT in middle-aged and old mice to the high levels observed in young mice. DHEAS, a major naturally occurring adrenal steroid that decreases in blood serum with age, could be rate-limiting in achievement of retention of learning. PMID- 2968831 TI - Off-road recreational motor vehicle accidents: hospitalization and deaths. AB - There is increasing concern over the unregulated use of recreational off-road motor vehicles. A review of 207 patients admitted to a tertiary care hospital over a 5-year period, as well as deaths due to use of recreational vehicles, elicited the following information. Recreational accidents predominantly involved men in their mid-twenties. Children younger than 16 years were more frequently involved in all-terrain vehicle (ATV) or dirt-bike accidents and constituted more than one-third of the total. There was a ninefold increase in ATV accidents over the study period, so that by 1985 ATVs were the primary cause of off-road injuries (52%). The musculoskeletal system was most frequently injured (66%) followed by the head and face (25%). There was permanent disability in 10.6%, and 33% of the recreational deaths were in children younger than 16 years. The inherent instability of ATVs was confirmed by the finding that in 60% of accidents the vehicle had rolled or flipped. Stricter licensing requirements should be implemented, and public education is required to draw attention to the danger of these vehicles, particularly to children. There is a need for proper safety equipment and driver training. The issue of vehicle design must also be addressed by the industries concerned. PMID- 2968833 TI - Ocular lesions in chickens inoculated with Escherichia coli. AB - Specific-pathogen-free chickens (two, four and ten weeks of age) which were inoculated via the air sac with Escherichia coli developed ocular lesions. Histologically, the main ocular lesions consisted of hyphema, hemorrhages of the iris, hypopyon, keratitis and uveitis. Hyphema was associated with hemorrhages of the iris, and hypopyon with keratitis and uveitis. Cyclophosphamide treatment enhanced the incidence and severity of hyphema and hemorrhages of the iris in the chickens. PMID- 2968832 TI - [Circulatory and respiratory effects of ketamine in horses anesthetized with halothane]. AB - Ketamine (0.3 mg/kg) administered intravenously to 12 halothane anesthetized horses caused a significant respiratory depression during ten minutes when respiration was spontaneous. Significant hemodynamic effects were not observed except for cardiac index. Clinical application was also discussed. PMID- 2968834 TI - Effect of intratracheal inoculation of Pasteurella haemolytica cytotoxin on the integrity of rat lung. AB - This study was conducted to investigate the in vivo effect of a single intratracheal inoculation of Pasteurella haemolytica cytotoxin on the rat lung. Changes in the biochemical and cytological composition of bronchoalveolar lavage fluid were used to estimate the magnitude of pulmonary cell injury, inflammatory response, vascular permeability and functional status of pulmonary alveolar macrophages. Effect of treatment was compared with rats intratracheally inoculated with supernatants of Pasteurella multocida or with sterile physiological saline solution (vehicle). Results indicated that Pasteurella haemolytica supernatants were not significantly toxic for the lungs of rats. PMID- 2968836 TI - Treatment of experimental ovarian carcinoma with monthly injection of the agonist D-Trp-6-LH-RH: a preliminary report. AB - Hormones, particularly gonadotropins, have been implicated in the development of ovarian cancer. Chronic administration of agonistic analogs of luteinizing hormone releasing-hormone (LH-RH) induces an inhibition of the pituitary-gonadal axis. The blockade of the release of luteinizing-hormone and follicle-stimulating hormone (FSH) may exert a possible therapeutic effect on ovarian cancer. We examined the results of prolonged administration of D-Trp-6-LH-RH, an agonistic analog of LH-RH in experimental ovarian cancer. We used the recently developed ovarian cancer model in rats, which is produced by treatment of pregnant rats with N-nitrosobis(2-oxopropyl)amine (BOP), following which a high incidence of ovarian tumors are induced in the offspring. In morphologic aspects the induced tumor resembles human ovarian neoplasms. Once a month administration of a delayed release preparation of microcapsules of D-Trp-6-LH-RH prolonged the survival and decreased tumor growth and the incidence of metastases. Additional experimental and clinical studies are needed to determine the efficacy of the treatment with LH-RH analogs in ovarian cancer. PMID- 2968835 TI - A study on the relationship between the pineal gland and the opioid system in patients with cancer. Preliminary considerations. AB - Recent studies showed that both the pineal gland and the endogenous opioid system are involved in the modulation of the immune system and in the regulation of tumor growth. Moreover, a relationship between pineal and opioid system has been demonstrated. In order get an overall view of the psychoneuroendocrine interactions in cancer patients, the levels of melatonin, the most important pineal hormone, and of beta-endorphin have been measured on blood samples collected during the morning. The study was carried out on 54 patients, 42 healthy subjects, and in 34 patients having illnesses other than cancer. Breast cancer, lung carcinoma, and colorectum cancer were the three neoplasms detected in the patients investigated. Growth hormone (GH), somatomedin-C and prolactin (PRL) levels were also determined. beta-endorphin levels were found to be substantially within the normal range in patients with cancer, whereas those of melatonin were raised in several cases. The beta-endorphin/melatonin ratio was higher than 2 in normal subjects, in non-neoplastic patients and in most cancer patients without metastases, whereas this ratio was lower than 2 in almost all patients in a metastatic stage of the disease. Neither melatonin levels nor those of beta-endorphin appeared to be significantly correlated with GH, somatomedin-C, and PRL concentrations. The low beta-endorphin/melatonin ratio observed in metastatic patients suggests the presence of an unbalanced relation between the pineal and the opioid system in those subjects. Therefore, an anomalous relationship between pineal function and opioid activity might play a role in the clinical course of neoplastic disease. PMID- 2968837 TI - Detection of haematoporphyrin derivative and haematoporphyrin excited states in cell environments. AB - The photosensitiser currently used in the photodynamic therapy of cancer is haematoporphyrin derivative. Pulsed laser studies of this material and also of the "parent" molecule haematoporphyrin in polyoma-transformed fibroblast cells have now been studied. We report, for the first time, the observation of the triplet absorption of these sensitisers in cells. The corresponding triplet triplet spectra are red shifted compared to aqueous buffer, lambda(max) 420 nm shifts to 460 nm. We also report our failure to observe singlet oxygen from the cells even though the triplet state can be seen to interact with the oxygen and even though singlet oxygen can be readily detected with the same sensitisers bound to serum albumin. PMID- 2968839 TI - Transluminal balloon angioplasty of infrahepatic caval anastomotic stenosis following liver transplantation: case report. AB - Treatment of an infrahepatic caval anastomotic stenosis with percutaneous transluminal balloon angioplasty is described in a patient 5 weeks after liver transplantation. Pressure measurements confirmed the significance of the obstruction and the success of the dilatation procedure. PMID- 2968838 TI - Increased levels of DNA lesions induced by leucovorin-5-fluoropyrimidine in human colon adenocarcinoma. AB - Leucovorin augments the growth inhibitory effect of 5-fluoropyrimidines on neoplastic cells. The effect is paralleled by much higher levels of DNA fragmentation than in cells treated with 5-fluoropyrimidines alone at the same concentration. The lesions are induced by a mechanism independent of incorporation of the drug into DNA, in all probability due to reduced repair of DNA lesions induced independently of the drug treatment. Thymidine added after the treatment with fluoropyrimidines partly rescues the cells and reduces the level of DNA fragmentation. PMID- 2968840 TI - Unusual transplant renal angioplasty complication: case report. AB - We report an avulsion of a transplant upper pole renal artery following apparently successful percutaneous transluminal angioplasty (PTA) of a stenosis involving main, upper, and lower pole renal arteries. We believe that the use of a high pressure balloon was the causative factor. If the branches at the bifurcation cannot be protected and if the stenoses fail to resolve under moderate balloon insufflation pressures (less than 5-6 atmospheres), consideration should be given to surgical revascularization. PMID- 2968842 TI - Turning lambda Cro into a transcriptional activator. AB - According to our present understanding, lambda repressor bound to DNA stimulates transcription by touching RNA polymerase bound at an adjacent promoter. The part of repressor required for activation was identified in part by the isolation of mutants specifically impaired in transcriptional activation. The amino acids of repressor altered in these "positive control" mutants lie in an acidic patch on the surface of repressor that is closely apposed to RNA polymerase. In this study, we show that this "activating patch" of repressor is sufficient for transcriptional activation in another sequence context. We transfer this activating patch onto the surface of lambda Cro, a protein normally unable to activate transcription, and show that the modified Cro is a transcriptional activator. In addition, we provide evidence that the repressor protein of phage 434 also activates transcription using an activating patch similar to that of lambda repressor. PMID- 2968841 TI - Tangled roles: a nurse practitioner's experience as a cardiac surgery patient. PMID- 2968844 TI - The pharmacologic regulation of interleukin-1 production: the role of prostaglandins. AB - The role of prostaglandins in the regulation of lipopolysaccharide (LPS)-induced interleukin-1 (IL-1) production by murine C3H/HeN resident peritoneal macrophages was studied. IL-1 production was initially studied in the presence of piroxicam and indomethacin, both inhibitors of prostaglandin biosynthesis. IL-1 was assayed using the IL-1-dependent proliferative response of C3H/HeJ thymocytes. LPS stimulation resulted in 15 to 20 ng/ml of prostaglandin E2 (PGE2) produced in the first hour of culture. IL-1-containing supernatants from drug-treated macrophages at dilutions of up to 1:32 resulted in enhanced thymocyte proliferation compared to control, non-drug-treated cultures and contained less than 2 ng/ml of PGE2. Similar enhancement of proliferation could be obtained by incubating non-drug treated supernatants with monoclonal anti-PGE2 but not anti-thromboxane B2 (TxB2) antibody. Further dilutions of the drug-treated supernatants gave thymocyte proliferation responses which were indistinguishable from control cultures and, correspondingly, had identical values for IL-1 production. The absence of an effect on IL-1 production was confirmed by quantitation of intracellular IL-1 alpha using goat anti-IL-1 alpha antibody and by quantitation of supernatant IL-1 receptor competition assay. Exogenous PGE2, in the concentration range produced in macrophage supernatants (10-20 ng/ml), directly inhibited IL-1-stimulated thymocyte proliferation. Finally, when macrophages were stimulated with LPS for 24 hr in the presence of added PGE2, thymocyte proliferation was inhibited at the lowest supernatant dilutions, but as the IL-1-containing supernatants were diluted out, the assay curves were indistinguishable from non-PGE2-treated control. Thus, in this system, PGE2 has no effect on IL-1 synthesis, but rather has a direct inhibitory effect on thymocyte proliferation. Nonsteroidal anti inflammatory drugs are not stimulating IL-1 production but are, in fact, relieving inhibition of the thymocyte IL-1 assay caused by the presence of prostaglandins. PMID- 2968843 TI - Subclass specificity of the Fc receptor for human IgG on K562. AB - The erythroleukemic cell line K562 bears a 40-kDa Fc receptor (Fc gamma RII) serologically related to and with a similar molecular weight as the Fc gamma R present on a broad range of leukocytes. The human IgG subclass specificity of the Fc gamma R on K562 was investigated using IgG aggregates of defined size, obtained from purified human myeloma proteins. The monoclonal antibody IV.3, which reacts with the Fc gamma RII present on various cell types, totally prevented binding of 125I-IgG2 trimers to K562. Experiments with radiolabeled IgG2 trimers showed that K562 cells bound a mean of 156,764 +/- 9895 molecules per cell with an association constant (Ka) of 1.8 +/- 0.7 X 10(8) M-1. Similar results were obtained with IgG3 oligomers. IgG3 and IgG2 trimers were about two- to threefold more effective in inhibiting binding of 125I-IgG2 trimers to K562 than IgG1 and IgG4 trimers. These results were confirmed by inhibition experiments using IgG monomers. The subclass specificity of the Fc gamma RII on K562 (i.e., IgG2 = IgG3 greater than IgG1 = IgG4) is quite distinct from the one reported for the Fc gamma RI and III of human cells (i.e., IgG1 = IgG3 greater than IgG4 and IgG2). PMID- 2968845 TI - The S-100 beta protein in normal human peripheral blood is uniquely present within a discrete suppressor-T-cell compartment. AB - The S-100-positive T lymphocytes, and, particularly, the S-100 beta subunit, are restricted, as demonstrated by quantitative subset analysis and double-labeling (gold-peroxidase) immunoelectron microscopy of T-cell subpopulations, to an unique T8-positive cell subset which interestingly was 9.3-negative and CD11b positive. Since both the T8-positive, 9.3-negative and the T8-positive, CD11b positive subpopulations have been demonstrated to show suppressive activities, the S-100-positive T cells seem to be closely restricted to a small T-suppressor cell compartment. Although functional studies on viable isolated S-100 beta positive cells are impossible to achieve, due to the lack of this protein on the cell membrane, its presence in a discrete T-suppressor compartment might suggest a possible role for the S-100 beta-positive T cells in the regulation of the immune system. PMID- 2968846 TI - Age-related decline in the lethal hit but not the binding stage of cytotoxic T cell activity in mice. AB - The activity of allogeneic cytotoxic T lymphocytes (CTL) declines with aging, as measured at the population level in chromium release assays, and is associated with a decreased proliferative response to the inducing alloantigens. At the single cell level, no difference was noted between target binding cells obtained from alloantigen-stimulated spleen cells of young and old mice. However, a consistent, large, and significant difference was observed between CTL generated from the spleens of young and old mice in the percentage of bound effector cells with lysed targets, i.e., the percentage of killer cells. These results show age related changes in the lethal hit stage and not the binding stage of CTL activity. This change could be used as a probe to identify the important mechanism(s) which mediates CTL activity. PMID- 2968847 TI - An AIDS update. PMID- 2968848 TI - Keeping abreast of dental news nationwide. PMID- 2968850 TI - Low back pain in the competitive tennis player. AB - The etiologies of low back pain and the biomechanics and pathology of the lumbar spine as they relate to tennis stroke mechanics have been reviewed, and a treatment protocol has been presented. A recent survey of the Men's Professional Tennis Tour is the only article found that discusses low back pain in tennis players; the orthopaedic and sports medicine literature is otherwise devoid of any relevant studies. Because this one survey indicates that 38 per cent of 143 tennis players missed at least one tournament because of low back problems, it seems obvious that an epidemiologic study on low back pain in racquet sports is vital to a more thorough understanding of the problem. PMID- 2968849 TI - Thoracoabdominal musculoskeletal injuries in racquet sports. AB - Thoracoabdominal musculoskeletal injuries are separated into rectus abdominus injuries and lower thoracic muscular injuries. Localized rectus injuries may be treated by steroid injection. Diffuse rectus injuries and thoracic injuries are treated by aggressive rehabilitation. The mechanisms of injury are different in each case, and prevention of these forces is necessary to allow soft-tissue healing. PMID- 2968851 TI - Abdominal and groin injuries in tennis. AB - Although abdominal and groin injuries are not unique to the game of tennis, the very mechanics of the ground stroke and overhead volley predispose participants in this sport to the chronic pain and frustration of these injuries. Careful consideration must be given not only to the muscular anatomy of the region but also to the visceral and neurologic anatomy. A patient, controlled program of rehabilitation, emphasizing flexibility and subsequent strengthening, must be stressed if recurrence is to be avoided. PMID- 2968852 TI - Preparation of a cell-free translation system from a wild-type strain of Neurospora crassa. AB - We describe the preparation of an in vitro translation system from a wild-type strain of Neurospora crassa. The system is capable of supporting efficient and faithful translation of native and in vitro transcribed eukaryotic messages. The translation products have minimal background and can be clearly analyzed by SDS polyacrylamide gel electrophoresis. The method of preparation of the lysate is simple, fast and reproducible. The procedure should be readily applicable to other filamentous fungi. PMID- 2968853 TI - [Screening of active anti-inflammatory-immunosuppressive and antifertile compositions from Tripterygium wilfordii. I. Screening of 8 components from total glucosides of Tripterygium wilfordii (TII)]. PMID- 2968854 TI - [Screening of active anti-inflammatory-immunosuppressive and antifertile components from Tripterygium wilfordii. II. Screening of 5 monomers from total glucosides of Tripterygium wilfordii (TII)]. PMID- 2968855 TI - [The immunoregulation of IgE antibody production in mice. IV. Anti-ovalbumin IgE antibody response]. PMID- 2968857 TI - [Nucleic acid hybridization analysis of the regulatory effect of a cytoplasmic factor on the malignancy of myeloma cells. I. Beta-globin gene expression]. PMID- 2968856 TI - [Establishment of a transplantable human lung adenocarcinoma cell line in nude mice and observation of its biological and ultrastructural characteristics]. PMID- 2968858 TI - [PAGE electrophoresis of hemoglobin from reticulocytes of myeloma cell hybrids]. PMID- 2968859 TI - [Studies on glucose metabolism and local cerebral blood flow in bicuculline induced rat epilepsy]. PMID- 2968861 TI - [Preparation of anti-VIP rabbit antiserum and its characterization]. PMID- 2968862 TI - [Pulmonary vascular compliance and distribution in dogs]. PMID- 2968860 TI - [Comparison of the labelling effects of CB-HRPs prepared with domestic and foreign materials]. PMID- 2968863 TI - [Assay procedures for the determination of monoamines in the rat and mouse brain by means of reversed-phase high-performance liquid chromatography with electrochemical detection]. PMID- 2968865 TI - [Effects of nifedipine and anisodamine on platelet membrane fluidity and platelet aggregation in rats]. PMID- 2968866 TI - [Synthesis of D,L-15-methylprostacyclin sodium salt]. PMID- 2968864 TI - [Retinoic acid and its analogs enhance allogeneic CTL induction in mice]. PMID- 2968867 TI - [Suppression of human B cell proliferation by PGE]. PMID- 2968868 TI - [Procedures of fractional isolation of fragment beta from tetanus toxin (T beta) and a coupling preparation of T beta-HRP]. PMID- 2968869 TI - Quantification of DNA hybrids on discs of nitrocellulose filter. PMID- 2968870 TI - Changes in vascular vasopressin receptors and responsiveness in DOCA/NaCl-treated rats. AB - This study investigated the effects of treatment with desoxycorticosterone acetate and salt (DOCA/NaCl) on blood pressure, aortic vascular vasopressin receptors and in vitro vascular responsiveness in male rats. Four groups of animals were utilized in the study: a control group on normal tap water, a control group drinking a 1% NaCl solution; a DOCA/NaCl-treated group (1% NaCl in the drinking water); and a uninephrectomized (1K) DOCA/NaCl-treated group. DOCA/NaCl treatment for 4 weeks resulted in a significant elevation in blood pressure which was more pronounced in the uninephrectomized animal. The concentration of specific binding sites for vasopressin on the aorta were reduced in both the DOCA/NaCl-treated groups. However, the vascular responsiveness of the aorta to vasopressin was significantly reduced only in the hypertensive, uninephrectomized-DOCA/NaCl-(1K DOCA/NaCl) treated rat. Both the maximal contraction and the sensitivity was reduced in the 1K DOCA/NaCl group. The results of this study would suggest that the vascular alterations to vasopressin are probably post receptor mediated and result from the DOCA/NaCl-induced hypertension. PMID- 2968872 TI - Serial study of T-lymphocyte subsets in women during very early pregnancy. AB - The aim of this serial study was to define variations in T-lymphocyte populations during the first weeks of pregnancy. Results obtained with 14 women pregnant after artificial insemination show that the CD4+ population decreased significantly as soon as the fourth week. This decrease was concomitant with the HCG peak and could be compatible with certain immunological mechanisms participating in tolerance to the fetal graft. PMID- 2968871 TI - In vitro activation of T lymphocytes from HIV-seropositive blood donors. II. Decreased mitogen-induced expression of interleukin 2 receptor by both CD4 and CD8 cell subsets. AB - Mononuclear cells (MC) from many individuals exposed to human immunodeficiency virus (HIV) exhibit a reduced proliferative response to a suboptimal concentration of phytohemagglutinin (PHA). However, the relative contributions of the 2 major T-cell subsets, namely CD4 and CD8 lymphocytes, to this reduced response remain unclear. Based on reports that interleukin 2 receptor (IL2R) expression correlates well with proliferative responses in HIV infection, we used dual-color cytofluorometry to measure IL2R expression by CD4 and CD8 cells following PHA activation of MC from HIV-seropositive blood donors. For data analysis, this study group was divided into two subgroups on the basis of DNA synthesis responses (seropositive with normal DNA synthesis, designated sero + NML, or seropositive with decreased DNA synthesis, designated sero + LOW). When compared to the seronegative control and sero + NML groups, the sero + LOW group exhibited significant reductions in the percentage of MC expressing IL2R, the proportion of CD4 cells expressing IL2R, and the proportion of CD8 cells expressing IL2R. In contrast, these parameters were unchanged in the sero + NML group compared to the control group. These findings show that reduced PHA-induced proliferative responses by MC from HIV-infected persons are associated with decreased IL2R expression by both CD4 and CD8 lymphocyte subsets. PMID- 2968873 TI - Expression of Fc epsilon receptors and surface and cytoplasmic IgE on human fetal and adult lymphopoietic tissue. AB - The appearance during ontogeny of IgE-positive and Fc epsilon receptor (FcER) bearing cells was studied. Monoclonal antibody to the constant region (Fc) of IgE (CIA-E-7.12) was used to detect cytoplasmic and surface IgE. A monoclonal antibody to the low-affinity Fc epsilon receptor (FcER-II = CD23) and immune complexes composed of human IgE and mouse monoclonal anti-human IgE Fc were used to detect FcER. Cryostat sections of human fetal tissues (liver, lung, spleen, and thymus) from 11 to 22 weeks gestation as well as adult tonsil tissues were examined for IgE, FcER, and other lymphoid markers by immunoperoxidase staining. Although both IgE- and FcER-positive cells were present in adult tissues, we found that, in contrast to an earlier report, such cells were not present in the fetal tissues examined. The in situ location of FcER on cells in human lymphoid tissues revealed that the FcER-bearing cells were localized predominantly in the germinal centers (mature B cell and macrophage areas) of the tonsil follicles with some staining in the mantle (resting and less mature B cell areas). PMID- 2968874 TI - Scintigraphic detection of clavicular hyperostosis in a patient with fulminant acne. AB - Intense Tc-99m MDP uptake in both clavicles was noted in a patient with fulminant acne. X-ray examination and biopsy performed later showed hyperostosis. PMID- 2968877 TI - Metformin revisited: its actions and indications for use. PMID- 2968875 TI - Pharmacokinetics and absorption of foscarnet after intravenous and oral administration to patients with human immunodeficiency virus. AB - Six patients with human immunodeficiency virus were given foscarnet in oral solution, 4000 mg every 6 hours for 3 days, followed by a washout period for 2 days and continuous intravenous infusion of 16,000 mg/24 hr over 72 hours. After oral foscarnet, plasma concentrations were less than 33 mumol/L in four patients; two had occasional concentrations of 35 to 50 mumol/L. The extent of absorption varied between 12% and 22%. During intravenous infusion, plasma concentrations ranged between 75 and 265 mumol/L. The disposition of foscarnet was triphasic, with mean half-lives of 0.45, 3.3, and 18 hours. Excretion data suggested elimination was by tubular secretion and glomerular filtration. Renal clearance was 176 ml/min 1.73 m2. The apparent nonrenal clearance, 40 ml/min 1.73 m2, probably reflects sequestration of foscarnet into bone. Ten percent to 28% of the cumulative dose may have been deposited in bone 2 days after infusion. A slight increase in serum calcium levels and changes in serum phosphate values may reflect the uptake of foscarnet in bone. Five patients had diarrhea (oral) and two had thrombophlebitis (intravenous). PMID- 2968876 TI - The OGTT: gold that does not shine. PMID- 2968878 TI - Diabetes-related perinatal mortality and congenital fetal abnormality: a problem of audit. PMID- 2968879 TI - Heart rate fluctuations in diabetic patients with cardiac vagal dysfunction: a spectral analysis. AB - The autonomic nervous control of cardiac function during active orthostatic load has been studied by measuring the power spectrum of heart rate fluctuations in 16 insulin-dependent diabetic patients and 14 age-matched control subjects. The patients were subdivided into two groups: 8 with normal respiratory sinus dysrhythmia (RSA+) and 8 with reduced respiratory sinus dysrhythmia (RSA-). In RSA- patients the total power (0.01-0.50 Hz) was significantly reduced compared with control subjects (4.7 versus 15.5 min-2, 2p less than 0.05) and the pattern of heart rate fluctuations was characterized by a relative increase in the low frequency component (0.01-0.05 Hz) as compared with RSA+ patients and control subjects (45% versus 24% and 27%, both 2p less than 0.01). There was also a significant reduction in the high-frequency component (0.15-0.50 Hz) as compared with RSA+ patients and control subjects (17% versus 36% and 33%, both 2p less than 0.05). During standing, a significant increase in total power was found only in control subjects (2p less than 0.01) and the difference between control subjects, and RSA+ and RSA- patients reached significance (32.2 versus 15.1 and 12.7 min-2, 2p less than 0.02 and 2p less than 0.01). The pattern of heart rate fluctuations in RSA- patients showed no significant change on standing. These results suggest that the reduced overall heart rate variability in diabetic patients with cardiac autonomic neuropathy is associated with a typical heart rate fluctuation pattern. PMID- 2968880 TI - Insulin requirement in non-insulin-dependent diabetes mellitus: relation to simple tests of islet B-cell function and insulin sensitivity. AB - Evaluation of simple tests of islet B-cell function and insulin sensitivity as predictors of metabolic control was performed during 3 months of insulin withdrawal in 25 insulin-treated diabetic subjects. All patients had a glucagon stimulated plasma C-peptide concentration above 0.33 nmol/l and a fasting plasma C-peptide concentration above 0.20 nmol/l a few days before insulin withdrawal. Insulin sensitivity was measured as the glucose disappearance rate (k) during an intravenous insulin tolerance test. Two patients were considered insulin requiring due to high fasting blood glucose levels (greater than 20 mmol/l) and two patients due to an increase in glycosylated haemoglobin of more than 1.1% (greater than approximately 3SD) in combination with weight loss. None of the remaining patients had a significant increase in glycosylated haemoglobin. An inverse correlation was found between stimulated C-peptide levels and insulin sensitivity (r = 0.41, p less than 0.05). Fasting and stimulated C-peptide concentrations of 0.40 and 0.70 nmol/l, respectively, separated non-insulin requiring patients from a group consisting of both insulin- and non-insulin requiring patients. At these C-peptide levels the predictive value of a positive test was 100% while the predictive value of a negative test was as low as 33% or 27% depending on whether fasting or stimulated C-peptide concentration was used. Including the k value in the prediction only increased the predictive values of negative tests to 40% and 33%, respectively. PMID- 2968881 TI - Limited joint mobility in the diabetic foot: relationship to neuropathic ulceration. AB - Limited joint mobility in the hand is a common manifestation of diabetes with the reported prevalence in insulin-dependent diabetes varying between 8 and 43%. Sixty-two subjects were studied in three groups (controls, diabetic patients without foot problems, and diabetic patients with neuropathic ulceration) to determine whether similar changes occur in the joints of the foot and to examine any possible relationship with neuropathic ulceration. There was a significant impairment of mobility in the range of motion of the sub-talar joint in diabetic patients with ulcers when compared with controls (p = 0.0001) or with the other diabetic patients (p = 0.004). There was a significant correlation between sub talar range of motion and mobility in other joints of the foot such as at the hallux (r = 0.59, p less than 0.001), or with mobility of the 5th finger (r = 0.41, p less than 0.01). There was also a significant association between the clinical presence of limited joint mobility in the hand, Dupuytren's contracture, and mobility of the sub-talar joint (p less than 0.05). Furthermore, impairment of mobility of the sub-talar joint was greatest on the affected side in those diabetic patients with neuropathic ulceration (p = 0.029). We conclude that the syndrome of limited joint mobility also affects the joints of the feet of diabetic patients and may predispose to ulceration in susceptible neuropathic feet. PMID- 2968882 TI - Glycosylation and cross-linkage of cardiac myosin in diabetic subjects: a post mortem study. AB - We have investigated the possibility that post-translational modification of myosin by protein glycosylation and cross-linking occurs in cardiac myosin. Left ventricular muscle was obtained at post-mortem from 6 diabetic and 7 non-diabetic subjects. Myosin was extracted from muscle and purified using Sephadex chromatography followed by protein concentration. Glycosylation was estimated using boronate affinity chromatography with the myosin dissolved in a pyrophosphate buffer, the glycosylated myosin being displaced with sorbitol. Cross-linkage was assessed by fluorescence at 440 nm upon excitation at 370 nm. Diabetic subjects had significantly higher levels (p less than 0.02) of glycosylated myosin (median 6.0% (range 3.8-6.6%] than non-diabetic subjects (median 2.4% (range 0.3-4.2%] but there was no difference in the degree of cross linkage as assessed by fluorescence (diabetic median 9.8 (range 6.5-17.0) arbitrary units; non-diabetic median 9.7 (range 6.0-11.4) arbitrary units). Glycosylation of left ventricular myosin may be of relevance to the excess risk of congestive cardiac failure in diabetic patients. PMID- 2968883 TI - Microalbuminuria in diabetes: a population study of the prevalence and an assessment of three screening tests. AB - A single observer reviewed 842 of the 917 known diabetic patients registered with 40 GPs in the Poole area. A midstream urine specimen was tested for proteinuria using Albustix (Ames) and cultured to detect bacterial infection. After the first 3 months of the survey, the aliquot of this specimen was frozen for later determination of the random albumin/creatinine ratio (R-Alb/Creat). Patients were requested to submit a timed overnight urine collection for estimation of urinary albumin excretion rate (AER). Of the 842 patients reviewed, 493 (59%) submitted timed overnight urine collections; 43 were excluded because of urinary infection and/or proteinuria. One hundred and thirty-three (30%) of 450 diabetic patients were found to have microalbuminuria, although only 31 (7%) had an AER greater than 30 micrograms/min. Six hundred and seven urine samples were collected for R Alb/Creat but 68 were excluded because of infection and/or proteinuria; in 10 further samples urinary creatinine was not measured. Two hundred and four (38%) of 532 diabetic patients were found to have an elevated R-Alb/Creat. There was a significant correlation between AER and R-Alb/Creat (r = 0.32, p less than 0.001) but a considerable number of patients showed either a normal AER and high R Alb/Creat or the reverse. The value of R-Alb/Creat or an overnight urinary albumin concentration, or an overnight urinary albumin/creatinine ratio (ON Alb/Creat) as screening tests to predict AER greater than 30 micrograms/min was assessed. An ON-Alb/Creat greater than 2.0 mg/mmol was the optimal screening test (sensitivity 96% and specificity 99.7%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968884 TI - Microalbuminuria in diabetes: relationships between urinary albumin excretion and diabetes-related variables. AB - A single observer reviewed 842 of the 917 known diabetic patients registered with 40 GPs in the Poole area. Fifty-nine per cent (493) of those reviewed submitted a timed overnight urine collection to measure albumin excretion rate (AER) and overnight albumin/creatinine ratio (ON-Alb/Creat); 43 samples were excluded because of urinary tract infection and/or proteinuria. A random urine sample was obtained in 607 diabetic patients to measure the random albumin/creatinine ratio (R-Alb/Creat); 68 specimens were excluded because of infection and/or proteinuria, and in a further 10 samples urinary creatinine was not measured. Stepwise multiple regression analyses found significant associations with the following variables: for AER, blood glucose (p = 0.001), smoking category (p = 0.002), sex (p = 0.034), and systolic blood pressure (p = 0.035); for R Alb/Creat, blood glucose (p = 0.001), retinopathy (p = 0.004), systolic blood pressure (p = 0.004), diastolic blood pressure (p = 0.015), coronary artery disease (p = 0.02), sex (p = 0.034), and vibration sense (p = 0.038). Interestingly, glycosylated haemoglobin was not a significant determinant of albuminuria in either analysis. PMID- 2968886 TI - Serum IGF-1 concentration in diabetic retinopathy. AB - Growth factors such as growth hormone and insulin-like growth factor 1 (IGF-1) may be important in the pathogenesis of diabetic retinopathy. We measured serum IGF-1 in 371 diabetic patients attending a diabetic retinopathy clinic and in 73 non-diabetic control subjects. No significant difference was observed in IGF-1 level between the diabetic and control groups (168 +/- 3.9 vs 177 +/- 7.4 micrograms/l [mean +/- SE]). Within the diabetic group, there was no difference between patients with no retinopathy and those with proliferative change (198.7 +/- 8.8 vs 190.5 +/- 11 micrograms/l). After adjusting for differences in age, duration of diabetes, and presence of proteinuria, only the inactive previously proliferative group showed any significant difference from the other patient subgroups (151.8 +/- 11.5 micrograms/l; p less than 0.05). Serum IGF-1 correlated with age in the control group (r = 0.49; p less than 0.001) and to a lesser extent in the diabetic group (r = -0.23; p less than 0.05). IGF-1 levels were higher in patients with proteinuria than in those without proteinuria (196.8 +/- 10.3 vs 138.8 +/- 4.4 micrograms/l; p less than 0.001). PMID- 2968885 TI - A comparison of morning and bed-time ultralente administration when using multiple injections in adolescence. AB - Ten adolescents took part in a randomized sequential crossover study comparing two insulin regimes: (1) three preprandial injections of soluble insulin from a pen injector plus ultralente given before bed; (2) morning soluble insulin with ultralente, plus two preprandial injections of soluble. Nine patients completed the study. Metabolic profiles were performed at the end of each 3-month treatment period. Glycosylated haemoglobin levels did not fall significantly on either regimen, and there were no differences between the changes in HbA1 concentration on the two regimens. However, blood glucose concentrations tended to be lower with bedtime than with morning ultralente, and were significantly different at 0800 h (13.1 +/- 2.3 mmol/l vs 17.9 +/- 1.9 mmol/l, p less than 0.02) and 1000 h (14.5 +/- 1.4 mmol/l vs 18.6 +/- 0.7 mmol/l, p less than 0.03). Plasma free insulin concentrations were not different. Total daily insulin dose and the proportion of ultralente given in each regimen did not differ. Results indicate that the rise in early morning fasting blood glucose concentrations was less when ultralente was given at bedtime rather than in the morning. PMID- 2968887 TI - Progression of proteinuria in type 1 and type 2 diabetes. AB - A longitudinal study evaluating the time course of the transition from normal to microalbuminuria, and then on to macroalbuminuria, was made over a mean period of 7 years in a cohort of 52 patients with Type 1 diabetes and 61 patients with Type 2 diabetes. Transient episodes of micro- and macroalbuminuria were often observed before the ultimate development of persistent Albustix-positive proteinuria. The transition from normal to microalbuminuria and from micro- to macroalbuminuria was characterized by rises in renal albumin clearance accompanied by lesser rises in total proteinuria. Seven patients with Type 1 and 12 with Type 2 diabetes showed evidence of progression, the interval for the transition from normal to macroalbuminuria varying from 3 to 5 years. In Type 1 diabetic patients, the development of micro- and macroalbuminuria was associated with a decline in renal function and a rise in systolic blood pressure without a significant change in blood glucose control. In Type 2 diabetic patients, the development of microalbuminuria was associated with a small decline in renal function but no change in blood pressure or blood glucose control. It is concluded that the transition from normal to micro- and on to macroalbuminuria may be more rapid then previously reported and varies considerably among individuals. PMID- 2968888 TI - Excess body fat distribution and glucose homeostasis in obese Arab women. AB - The relationship of body fat distribution to glucose intolerance and non-insulin dependent diabetes mellitus in Arab women was studied in 102 obese non-diabetic and 40 obese women with diabetes. The obese women underwent a glucose tolerance test. Linear regression analysis revealed a significant correlation between the waist/hip ratio and the plasma glucose concentration at 120 min. When divided into two groups according to the median of their waist/hip ratio (0.815), obese women without history of diabetes but with high waist/hip ratio (0.86 +/- 0.07, mean +/- SD) had significantly higher prevalence of glucose intolerance and of diabetes mellitus than those with the low ratio (0.78 +/- 0.03, chi 2 = 9.32, p less than 0.001). The highest ratio (0.89 +/- 0.06) was observed in the obese women with known diabetes mellitus. PMID- 2968889 TI - District Diabetes Centres in the United Kingdom. A report on a workshop held by the Diabetes Education Study group on behalf of the British Diabetic Association. AB - Dissatisfaction with standards of diabetes care in general, and the traditional diabetic clinic in particular, is widespread. Problems include the large numbers of attenders, short consultation times (often with inexperienced medical staff), lack of continuity and prolonged waiting times. Standards of education and control fall woefully below those that are desired. In recent years several different strategies have been adopted to provide solutions, including general practitioner cooperative care schemes to reduce numbers and improve community care, the appointment of diabetes specialist nurses to take on the major educational role, and the commissioning of special education units designed to remove the educational element from the traditional clinic environment. Lately consideration has been given to the development of Diabetes Centres, to provide more comprehensive diabetes care, both educational and clinical. A workshop was held in January 1987 to analyse in greater detail the concept of Diabetes Centres and the consequences of their implementation. PMID- 2968890 TI - Knowledge profile and control in diabetic patients. AB - Knowledge about diabetes was assessed using a previously described interactive computer-based questionnaire in 79 patients with insulin-dependent (IDDM) and 72 with non-insulin-dependent (NIDDM) diabetes mellitus routinely attending a single diabetic clinic. Simple linear correlation of total knowledge score with glycosylated haemoglobin (HbA1c) showed no significant relationship for either IDDM (r = 0.12: p = 0.18) or NIDDM (r = 0.15: p = 0.1). However, quintile grouping of knowledge scores showed the mean HbA1c to be significantly higher in the lowest scoring NIDDM quintile (10.6 +/- 0.5: +/- SE) with respect to the pooled mean of all the higher scoring quintiles (9.0 +/- 0.3) (p = 0.027). Mean HbA1c (9.6 +/- 0.5) was also higher in the least knowledgeable IDDM quintile than any other quintile group (range 8.8-9.0) but this was not significant with respect to the pooled mean of higher scoring patients (p greater than 0.1). The mean age of the lowest scoring IDDM quintile group (60.5 +/- 13.9 years) was significantly higher (p less than 0.01) than higher scoring IDDM groups (mean age range 36.5-43.3 years) but age was not significantly related to HbA1c in IDDM subjects. IDDM showed greater knowledge of diabetes than NIDDM but ignorance in key areas was unacceptably high in both diabetic subtypes, indicating that regular knowledge assessment and educational reinforcement may be essential for good diabetic control as well as patient safety, particularly in older IDDM patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968891 TI - The nephrotic syndrome at presentation of insulin-dependent diabetes mellitus; cause or coincidence? AB - Two young women developed the nephrotic syndrome within 2 weeks of presenting with diabetes and starting insulin. One had a renal biopsy which showed changes consistent with 'minimal change nephrotic syndrome' on electron microscopy but no evidence of diabetic glomerulosclerosis. Neither patient received steroids; in one the oedema resolved spontaneously but the other required diuretics. This patient also had severe IgA-deficiency probably associated with epilepsy and/or phenytoin therapy and unrelated to the pathogenesis of the nephrotic syndrome. The nephrotic syndrome may rarely present coincidentally with, or soon after, insulin-dependent diabetes mellitus (IDDM). It must be distinguished from 'insulin oedema' and classical diabetic nephropathy which occurs later in the course of IDDM. All reported cases have either remitted spontaneously or responded to steroids. PMID- 2968892 TI - Diabetic neuropathic bladder associated with clinical features of iliofemoral venous thrombosis. AB - A 60-year-old female diabetic patient with autonomic neuropathy gradually developed a warm swollen left leg over a period of 12 days. She was found to have chronic retention of urine due to a neuropathic bladder. Venography did not reveal thrombus but showed compression of the ipsilateral external iliac vein by the bladder. No other cause for her warm swollen leg was found. Following the introduction of a bladder catheter, her leg gradually returned to normal. PMID- 2968893 TI - Pneumococcal osteomyelitis and cellulitis in an adult patient with diabetes mellitus. AB - A 48-year-old diabetic patient was admitted to hospital with fever and extensive infection of the tissues around the right elbow. Diagnosis was made of cellulitis and underlying osteomyelitis caused by Streptococcus pneumoniae. Although seen in patients with seriously impaired host defence mechanisms pneumococcal osteomyelitis and cellulitis has not been reported in a diabetic patient. PMID- 2968895 TI - The neurology of diabetes mellitus in Central Africa. AB - Six-hundred African diabetic patients were examined; 31% had peripheral neuropathy and 2% autonomic neuropathy. They were older and had had diabetes longer (p less than 0.001) than those without neuropathy. Blood glucose control, based on HbA1 levels, was poorer (p less than 0.05) in patients with neuropathy. A control group of 469 non-diabetic Africans was also examined. They were age- and sex-matched with the diabetic patients aged 35 years or more. Seven per cent had evidence of peripheral neuropathy, significantly less (p less than 0.001) than the diabetic population. Impotence was commoner in diabetic men than controls, mainly in association with neuropathy and the use of hypotensive drugs. PMID- 2968897 TI - British Diabetic Association Medical and Scientific Section spring meeting. 21-23 April 1988, Exeter. Abstracts. PMID- 2968894 TI - My sweetness problem. PMID- 2968896 TI - The Betalike: a practical artificial pancreas? PMID- 2968899 TI - Long-term care services in Costa Rica. PMID- 2968898 TI - Photodynamic therapy of cancer. AB - PDT represents another modality for the treatment of human malignancy. Photoactivated hematoporphyrins have definite antitumor activity in both in vitro and in vivo experimental systems. Much of the early clinical work involved treatment of patients with advanced, recurrent disease who had not responded to conventional therapy. Because good responses with acceptable toxicity have been obtained in these patients, active investigation continued and is aimed at defining the most appropriate sites and applications for the technique. Because of the limited depth of light penetration in tissue, the most promising sites may be those where there is limited thickness of tumor, such as in superficial skin lesions or carcinomas in situ involving the aerodigestive tract, bronchial tree, or genitourinary tract. Other potential uses include those where PDT could be combined with surgical or chemotherapeutic debulking, such as pleural mesothelioma or advanced stage ovarian cancer. Whether PDT can be of benefit in surgical cases where the margins of resection are close is an interesting but speculative notion at the present time. Clinical trials with hematoporphyrin derivative PDT in the sites mentioned are in progress. Laboratory work to better understand HpD also continues, as well as investigations into alternative photosensitizers with improved tumor localization, less cutaneous photosensitivity, and absorption peaks at deeper penetrating wavelengths of light. Attempts at measuring singlet oxygen, if successful, will permit the development of more meaningful dosimetry in order to correlate response with actual tissue levels of the purported cytotoxic agent. Hopefully, these and other developments in the field of PDT will improve the treatment for patients with cancer. PMID- 2968900 TI - [Superoxide dismutase in ischemic phenomena in the human. Obstruction by strangulation and cutaneous flaps]. PMID- 2968902 TI - [Protective action of nicotinamide and pyridoxine on the initiation stage of carcinogenesis induced in mice by procarbazine]. AB - It is shown that at the initiating stage of procarbazine carcinogenesis in F1 female mice the parenteral administration of nicotinamide or pyridoxine results in a significant decrease in the lung adenoma rate from 77% to 18 or 46%, respectively. Pyridoxal, pyridoxal-5'-phosphate and L-penicillamine did not influence the lung adenoma frequency. PMID- 2968901 TI - Immune status in healthy relatives of patients with familial Crohn's disease. AB - Immune-mediated mechanisms and genetic factors are believed to be involved in the pathogenesis of Crohn's disease. We studied T- and B-cell subpopulation proportions and various functional assays, including proliferative responses to PHA and Con A, Con A-induced suppressive activity, and natural killer cell assay toward the K562 cell line, in the peripheral blood of 22 patients with inactive familial Crohn's disease and their 35 healthy relatives including nine families. HLA-A, -B, and -DR antigens were determined in all the subjects. With the exception of minor abnormalities of suppressor cell activity present in some relatives of two families, neither significant impairments of immunological parameters in patients or their relatives nor concordant segregation of HLA haplotypes and disease were observed. These data indicate that peripheral immune abnormalities previously described in patients with Crohn's disease do not constitute primary factors involved in the disease itself and that familial incidence in Crohn's disease cannot be linked to immunological markers presently studied. PMID- 2968903 TI - [Effect of electron-acceptor compounds on the immunoregulatory mechanisms and ATPase activity of the erythrocyte membranes from patients with inoperable lung cancer]. AB - In 55 inoperable patients with lung cancer the use of tinidizole and metronidazole with X-ray therapy intensified the immunoregulative disbalance, increased the quantity of circulative immune complexes and reduced the Na+K+ ATPase activity of erythrocyte membranes in contrast to the data obtained from patients receiving only X-ray therapy. Thus the data obtained may be the basis for the use of the correcting therapy in inoperable patients with lung cancer. PMID- 2968904 TI - [Antitumor effectiveness of chalone-like proteoglycans isolated from normal and tumor tissues]. AB - Chalone-like proteoglycans (PG) have been isolated from 22a hepatoma, spontaneous adenocarcinoma of the mammary gland of C3H/He mice and from transplanted X-ray induced lymphoma in C57Bl mice. It has been found that the biological activity of these PG was considerably weaker than that of PG taken from the liver, mammary gland and spleen of cattle. PG from 22a hepatoma produced a weaker tissue specific antimitotic effect as against PG from the liver when acting on the liver cells of 6-day C57Bl mice, while the lymphoma PG under these conditions was ineffective. PG from the liver and mammary gland inhibited the growth of certain transplanted tumours in vitro. The lymphoma PG has stimulated the tumour growth. PMID- 2968906 TI - Biosynthesis of vitamin B-12 in anaerobic bacteria. Experiments with Eubacterium limosum and D-erythrose 14C-labeled in different positions. AB - In anaerobic microorganisms the origin of C atoms 2 and 4-7 of the 5,6 dimethylbenzimidazole moiety of vitamin B-12 is still unknown. In order to tackle this problem we added several 14C-labeled putative precursors to Eubacterium limosum fermentations. The degradation of the isolated vitamin B-12 revealed that only D-erythrose, 14C-labeled in different positions, was efficiently incorporated into the 5,6-dimethylbenzimidazole part. The 5,6 dimethylbenzimidazole obtained from an experiment with D-[U-14C]erythrose was further degraded. It was found that C-2 was unlabeled, whereas half of the label was located in C-5 plus C-6, and the other half in C-4 plus C-7. These results demonstrate that in E. limosum D-erythrose is a precursor of C-atoms 4, 5, 6 and 7 of the 5,6-dimethylbenzimidazole part of vitamin B-12. PMID- 2968905 TI - Insulin-like growth factor I receptors on mouse neuroblastoma cells. Two beta subunits are derived from differences in glycosylation. AB - We have characterized receptors for the insulin-like growth factor (IGF-I) on the mouse neuroblastoma cell line N18 as well as NG108, the hybrid cell line of N18 and rat glioma (C6). In this cell-free system, IGF-I and insulin stimulated the phosphorylation of 95-kDa and 105-kDa proteins. Using appropriate antibodies we were able to demonstrate that the IGF-I receptor beta subunit has two subtypes of 95 kDa and 105 kDa. On the other hand, insulin receptor beta subunit is a separate single 95-kDa protein. Enzymatic digestion of IGF-I receptor beta subunit subtypes by glycopeptidase F resulted in similar molecular masses (84 kDa and 86 kDa) on SDS-PAGE, which suggests that the difference in molecular masses between two subtypes is attributable to the differences in N-linked complex-type carbohydrate chains on the extracellular domain of beta subunits. This conclusion is further supported by peptides of similar molecular mass following staphylococcal V8 protease digestion. Analysis of IGF-I receptor beta subunit subtypes in these cells may provide insights into the mechanism of action of IGF I on neural tissues. PMID- 2968909 TI - Systolic and diastolic alterations of left ventricular function: prevalence in ischaemic heart disease and importance in the management of individual patients. AB - Myocardial perfusion and contractile performance are strictly interrelated. Conclusive experimental data show that changes in perfusion cause immediate changes in function, and conversely, that changes in contraction affect both total and regional myocardial blood flow. In ischaemic heart disease, both transient and permanent changes in global and/or regional left ventricular (LV) function have been described. In myocardial ischaemia, an impairment of LV relaxation, as indicated by a reduction of negative dP/dt, is the first appreciable haemodynamic event. When ischaemia is sustained, impairment of LV contraction follows, as indicated by a drop of positive dP/dt, reduction of systolic LV pressure, rise of end-diastolic LV pressure. Unrestricted reperfusion, as in experimental coronary occlusion and in Prinz-Metal angina, is associated with a transient rebound in regional systolic function. Spontaneous and/or induced changes of regional ventricular function provide relevant information for the management of the individual patient. In PTCA candidates monitoring regional contractile performance by mono and two-dimensional echocardiography during dypiridamole infusion has proven to be a sensitive and specific test for (1) assessing the haemodynamic significance of a coronary stenosis in patients with single-vessel disease, (2) identifying the 'culprit' lesion in patients with multiple-vessel disease. During PTCA, right and left ventricular function assessed by radionuclide angiography both at rest and during atrial pacing appears to be a sensitive method for the immediate evaluation of the efficacy of the procedure. PMID- 2968907 TI - Poor efficacy of teicoplanin in treatment of deep-seated staphylococcal infections. AB - Teicoplanin in a 400 mg intravenous loading dose followed by 200 mg/day intravenously or intramuscularly was given to 19 patients with deep-seated staphylococcal infections. Only eight patients (44.4%) were considered cured, failure mostly being observed in patients with osteomyelitis, endocarditis and bacteremia. Poor tissue kinetics of teicoplanin and the presence of foreign bodies are probable explanations for the reported failures. Future trials using a higher dose of teicoplanin with or without the addition of rifampicin or gentamicin seem to be justified. PMID- 2968908 TI - Heat treatment to increase phage typability of Staphylococcus aureus. AB - Heat treatment was used to reduce the number of Staphylococcus aureus strains that were not typable with the basic set of phages. All strains were phage typed according to the standard method after growth in broth at 37 degrees C or 48 degrees C. Forty-eight of 72 nontypable strains could be phage typed after heat treatment of the bacterial cultures. The page lysability increased with the higher incubation temperature of the broth, but the mean variability in the phage pattern of a strain was not significantly affected. The phage typing results of strains sampled over a period of several months were in accordance with the epidemiology, suggesting that phage typing after incubation at 48 degrees C is a stable and useful epidemiological tool. PMID- 2968910 TI - Left ventricular dysfunction, exercise capacity and activity recommendations. AB - There is lack of consistent correlation between left ventricular function at rest, haemodynamic measurements at rest, symptomatic status and exercise tolerance. Haemodynamic data appear comparable in patients whose exercise tolerance is limited clinically by fatigue and by breathlessness. Selected patients with left ventricular dysfunction and compensated congestive heart failure can safely undergo both exercise testing and training, and an improvement in functional status can be anticipated to result from exercise training. PMID- 2968912 TI - Biomechanics of the human spine and trunk. AB - This chapter has reviewed the past 30 years of experimental biomechanical studies of the spine and trunk. In the last 10 years, computers have allowed the development of simulation techniques and models to predict spine and muscle loading in most static and quasi-static activities. Some problems remain, however, particularly with activities involving bending and twisting and those that entail maximal efforts. Current research is focused on trying to validate models for the analysis of dynamic activities involving simple planar motions. Although the body segment kinematics and external support forces in complex motions can be measured fairly easily with modern motion analysis equipment, models that correctly predict the internal trunk forces have yet to be fully developed and validated. These models will be useful in studying how, why, and where failure of the soft and bony tissues is most likely to occur in a given activity, and whether it is related to work or athletics. The challenge for the future is to develop models that adequately reflect the anatomical sophistication of the spine and trunk. Thus the stress and strain distributions in any trunk musculoskeletal component, whether the posterior wall of the annulus, a muscle slip of the semi-spinalis group, or the lumbosacral endplate, will be able to be found. These results can then be combined with models of cumulative trauma response to successfully identify potential failure sites. PMID- 2968911 TI - PET measurement of D2 and S2 receptor binding of 3-N-[( 2' 18F]fluoroethyl)spiperone in baboon brain. AB - The regional pharmacokinetic behavior in baboon brain of 18F-fluoroethyl- and 18F fluoropropylspiperone (18FESP, 18FPSP) at specific activities greater than or equal to 1000 Ci/mmol was studied with PET. Four hours after injection of 5-10 mCi 18FESP, uptake in striatum was 0.048% +/- 0.005% of injected dose per cm3, which is almost the same as with 18F- and 11C-methylspiperone. While 18FPSP was taken up in much smaller amounts than 18FESP, striatum to cerebellum activity ratios were quite similar for both ligands (about 9 to 10 at 4 h p.i.). Because of its higher striatal uptake, 18FESP seems to be better suited for PET. Furthermore, relative binding to S2 receptors was much smaller for FESP: competing cold S2 antagonists (ritanserin, ketanserin) did not alter 18FESP binding to striatum, concurrently reducing uptake in frontal cortex by only 15% 20%. With coinjection of increasing amounts of cold FESP, saturation of 18FESP binding to striatum occurred at doses exceeding 10 micrograms per kg. Quantitative analysis of radiolabelled ligand in arterial plasma (decrease to 8% at 4 h p.i.) demonstrated identical metabolic turnover for both ligands. Direct use of binding fractions from the saturation curve resulted in overestimation of the receptor density in striatum. Using the 18FESP plasma concentration time curve and the dynamic uptake data, k3 of a three compartment model could be determined by non linear regression. However, dramatic changes of the dependence of k3 on the specifically bound ligand concentration were observed even at small loading doses of FESP. Estimation of Bmax yielded a D2 receptor density of only 6 pmol per cm3 in baboon striatum. PMID- 2968913 TI - Involvement of gangliosides and glycoprotein fibronectin receptors in cellular adhesion to fibronectin. AB - We have used a rat neural cell line, B65, to investigate the relative contributions of gangliosides and glycoprotein receptors in adhesion to fibronectin. Monoclonal antibodies against two neuroectoderm-associated gangliosides, D1.1 and GD3, inhibit the rate of B65 attachment to fibronectin, suggesting that these gangliosides are involved in the adhesion process. Adhesion to fibronectin is not affected by a third monoclonal antibody against a separate, unidentified cell-surface component of B65 cells. Furthermore, B65 cells lacking D1.1 adhere to fibronectin at a slower rate than B65 cells that express D1.1. The involvement of glycoprotein receptors in adhesion is demonstrated by the ability of antibodies against human fibronectin receptor to inhibit B65 attachment to fibronectin. In addition, adhesion is blocked by a hexapeptide containing the Arg Gly-Asp fibronectin sequence which is necessary for binding to the receptor. Trypsin treatment of B65 cells in the absence of divalent cations results in proteolysis of the fibronectin receptor with an accompanying loss of ability of the cells to attach to fibronectin. D1.1 and GD3 expression is not affected by this trypsinization, indicating that the gangliosides alone are incapable of mediating attachment. The glycoprotein receptors must be primarily responsible for adhesion to fibronectin with the gangliosides playing a secondary role as enhancers or modulators. PMID- 2968914 TI - Cell culture studies and oncogene expression in juvenile chronic myelogenous leukemia. AB - Juvenile chronic myelogenous leukemia (JCML) may be distinguished from adult CML based upon in vitro cell growth characteristics. We studied four untreated children with JCML and report additional unique findings. Peripheral blood (PB) and bone marrow (BM) cells were grown in soft agar. Without exogenous colony stimulating activity (CSA) there was exuberant "spontaneous" colony formation in both PB and BM cultures. In the absence of exogenous stimulus, PB colony morphology was predominantly, but not exclusively, monocyte/macrophage. When PB was depleted of adherent cells, "spontaneous" colony formation was nearly completely abrogated. Cultures were also performed in the presence of various sources of CSA including giant cell tumor-conditioned medium (GCT-CM), a melanoma cell line-CM (LD1-CM), human placenta-CM (HPCM), and normal PB mononuclear cell (PBMC) feeder layers. Colony formation was typically increased with HPCM and PBMC, whereas in two patients GCT-CM and LD1-CM failed to stimulate additional colony growth when compared to cultures without exogenous CSA and, in fact, appeared to inhibit baseline "spontaneous" growth. The morphology of colonies in the presence of exogenous stimuli was highly variable. Because of the recent association between the c-fms protooncogene product and the receptor for the monocyte growth factor CSF-1, we analyzed the PB cells from two JCML patients for c-fms expression. Although expressed, c-fms levels were less than that in an adult with Ph1-positive CML in chronic phase. These studies indicate that in JCML, there are dramatic increases in both PB and BM colony-forming cells and that "spontaneous" growth is dependent on an accessory adherent cell fraction. Furthermore, patterns of responsiveness to various sources of CSA suggest that the colony-forming cells may not be a uniform population of malignant cells. PMID- 2968915 TI - Radiation inactivation analysis of sarcoplasmic reticulum Ca-ATPase in membrane bound form and in detergent-solubilized monomeric states. AB - The sarcoplasmic reticulum Ca-ATPase was subjected to target size analysis by radiation inactivation in various buffer conditions and after solubilization in monomeric form in non-ionic detergent and in SDS. The target size was also determined for Ca-ATPase in bidimensional crystals formed in the presence of decavanadate or lanthanide. The standardization obtained with defined monomers of Ca-ATPase shows that the target size of Ca-ATPase in the functional membrane bound state may be ascribed to a single peptide chain, possibly with surrounding lipid. Further analysis of the radiation inactivation sizes of various partial reactions of the pump cycle, including phosphorylation and Ca2+ occlusion, indicated much smaller values than the target size pertaining to decomposition of the whole peptide chain. This is consistent with the existence of separate functional domains within a single peptide chain. PMID- 2968916 TI - Solubilization of immune precipitates by complement in the absence of properdin or factor D. AB - Various experiments have demonstrated that immune precipitates (IPs) are not solubilized by complement in the absence of alternative pathway function. To determine whether the characteristics of the IPs were responsible for these observations, we studied the solubilization (Sol) of IPs formed by bovine serum albumin (BSA)-rabbit antiBSA and tetanus toxoid (TT)-human antiTT. Sera deficient in properdin solubilized a fraction of BSA-antiBSA precipitates, although only when the IPs were formed in antibody excess. The same sera solubilized TT-antiTT precipitates with some delay but almost as efficiently as normal serum. Factor D depleted serum solubilized a fraction of TT-antiTT precipitates too, indicating that Sol may proceed through activation of the classical pathway only. Thus, the requirements for complement-mediated Sol depend on the characteristics of the IPs and do not necessarily include alternative pathway function. PMID- 2968917 TI - Evidence for phosphorylation of yeast phosphofructokinase. AB - Radioactively labelled material from yeast cells grown in the presence of [32P]phosphate was specifically recognized by antibodies raised against yeast phosphofructokinase. Purified yeast phosphofructokinase was phosphorylated in a cyclic AMP-independent manner by a protein kinase enriched from yeast extracts. This phosphorylation occurred specifically on the beta-subunit, and 0.56 mol of phosphate/mol of subunit was incorporated. The results indicate the phosphorylation of yeast phosphofructokinase both in vivo and in vitro. Phosphofructokinase phosphorylated in vitro was more stable against proteolytic degradation compared to the non-phosphorylated enzyme. PMID- 2968919 TI - The carboxy-terminal domain of the LexA repressor oligomerises essentially as the entire protein. AB - The ability of the isolated carboxy-terminal domain of the LexA repressor of Escherichia coli to form dimers and tetramers has been investigated by equilibrium ultracentrifugation. This domain, that comprises the amino acids 85 202, is readily purified after self-cleavage of the LexA repressor at alkaline pH. It turns out that the carboxy-terminal domain forms dimers and tetramers essentially as the entire LexA repressor. The corresponding association constants were determined after non-linear least squares fitting of the experimental concentration distribution. A dimer association constant of K2 = 3 X 10(4) M-1 and a tetramer association constant of K4 = 2 X 10(4) M-1 have been determined. Similar measurements on the entire LexA repressor [(1985) Biochemistry 24, 2812 2818] gave values of K2 = 2.1 X 10(4) M-1 and K4 = 7.7 X 10(4) M-1. Within experimental error the dimer formation constant of the carboxy-terminal domain may be considered to be the same as that of the entire repressor whereas the isolated domain forms tetramers slightly less efficiently. It should be stressed that the potential error in K4 is higher than that in K2. The overall conclusion is that the two structural domains of LexA have also well-defined functional roles: the amino-terminal domain interacts with DNA and the carboxy-terminal domain is involved in dimerisation reinforcing in this way the binding of the LexA repressor to operator DNA. PMID- 2968918 TI - cDNA sequence of the beta 2-subunit of human liver alcohol dehydrogenase. AB - A cDNA library of mRNA from a human liver expressing the beta 2-subunit of alcohol dehydrogenase was constructed in lambda gt11. One clone coding for 352 of a total of 374 amino acid residues of the beta 2-subunit was isolated. The sequence differed from that of the beta 1-subunit at one nucleotide position resulting in an Arg/His exchange at position 47 of the peptide chain, in agreement with data from protein sequence analysis [(1984) FEBS Lett. 173, 360 366]. PMID- 2968920 TI - Insulin-like growth factor I binding and receptor kinase in red and white muscle. AB - IGF-I receptors were partially purified from red and white skeletal muscle by lectin-affinity chromatography and the resultant fraction was depleted of insulin receptors by insulin affinity chromatography. Equilibrium binding of 125I-IGF-I to receptor preparations from red and white muscle yielded identical Scatchard plots. The integrity of the IGF-I receptor preparation in the two fiber types was identical as determined by affinity cross-linking. The tyrosine kinase activity of the receptor from red muscle was 2-3-fold more active towards exogenous substrates in both the basal and ligand-activated states as compared to white muscle. These data show that there is IGF-I-dependent kinase activity intrinsic to IGF-I receptors from skeletal muscle, and suggest that identical cellular factors may regulate the kinase activity of insulin and IGF-I receptors in a parallel manner in vivo. PMID- 2968921 TI - Different effects of two thromboxane A2/prostaglandin H2 receptor ligands, U46619 and S-145, on rabbit platelets. AB - Stimulation of rabbit platelets with U46619 induced platelet shape change, aggregation and secretion of ATP. However, S-145, which specifically binds to the thromboxane A2/prostaglandin H2 receptor like U46619, induced only shape change. Both compounds rapidly elevated cytoplasmic Ca2+ concentration although only U46619 evoked the formation of inositol phosphates. Chelating external Ca2+ with EGTA did not affect the S-145-induced platelet shape change while intracellular Ca2+ movement was severely reduced. These results suggest an essential role of phospholipase C in the induction of platelet aggregation and secretion and that some factor other than Ca2+ and phospholipase C participates in platelet shape change. PMID- 2968922 TI - Enzymatic activity and filament assembly of Acanthamoeba myosin II are regulated by adjacent domains at the end of the tail. AB - Polyclonal antibodies raised against a synthetic peptide consisting of the last 19 amino acids at the end of the coiled-coil region of the heavy chains inhibited the actin-activated Mg2+-ATPase activity of myosin II and its ability to form filaments. Antibodies against a synthetic peptide corresponding to the 21 adjacent amino acids at the beginning of the non-helical tailpiece, which include the three regulatory phosphorylatable serines, had no effect on either activity. PMID- 2968923 TI - Acidic fibroblast growth factor stimulates opsin levels in retinal photoreceptor cells in vitro. AB - It is demonstrated that newborn rat retinal photoreceptor cells can differentiate in monolayer culture, and synthesize de novo photoreceptor-specific proteins such as opsin. When maintained in serum supplemented medium on a laminin substrate, these cells survive for up to 3 weeks. The addition of acidic fibroblast growth factor stimulates an increase in the levels of opsin of 5-10-fold control values, and prolongs cell survival by up to 6 days. PMID- 2968924 TI - 1-Deoxymannojirimycin inhibits Golgi-mediated processing of glycoprotein in Xenopus oocytes. AB - We prepared in vitro an mRNA transcript coding for the erythroagglutinating subunit of the kidney bean glycoprotein phytohemagglutinin, E-PHA. The mRNA, injected into Xenopus oocytes, synthesized E-PHA carrying two Asn-linked carbohydrate chains, one of which was processed and acquired resistance to endo beta-N-acetylglucosaminidase H, as occurs in the native bean cells. When the mannose analog 1-deoxymannojirimycin, an inhibitor of mammalian Golgi mannosidase I, was included in the oocyte culture medium, the acquisition of endo-beta-N acetylglucosaminidase H resistance was abolished, indicating that also in an amphibian cell the inhibitor blocks a key reaction in Golgi-mediated processing. PMID- 2968926 TI - [Health education in preventing pustular skin diseases]. PMID- 2968927 TI - [Dorso-lumbar pain related to the handling of patients: how to avoid them?]. PMID- 2968925 TI - [Organization of sobriety corners in an industrial enterprise]. PMID- 2968928 TI - Monoclonal antibodies against human leucocyte antigens. II. Antibodies against CD45 (T200), CD3 (T3), CD43, CD10 (CALLA), transferrin receptor (T9), a novel broadly expressed 18-kDa antigen (MEM-43) and a novel antigen of restricted expression (MEM-74). AB - The specificities of several monoclonal antibodies described by us earlier were determined more exactly: (1) antibodies MEM-31 and MEM-32 are directed against CD8 and CD5 T cell antigens, respectively: (2) MEM-15 and MEM-18 react with the CD14 monocyte antigen; (3) MEM-28 recognizes the CD45 pan-leucocyte antigen. Several new monoclonal antibodies are described: (4) MEM-55 and MEM-58, reactive with subsets of CD45-related molecules; (5) MEM-43, recognizing a broadly expressed 18-kDa glycoprotein; (6) MEM-57, reacting with the CD3 antigen complex: (7) MEM-59 recognizing the CD43 leucocyte antigen; (8) MEM-74, which reacts with an unidentified antigen strongly expressed on several cell lines and many leukaemic cells but absent from most resting leucocytes; (9) MEM-75, directed against transferrin receptor, and (10) MEM-78, recognizing CD10 (CALLA) antigen. PMID- 2968930 TI - Incarcerated Richter's hernia after laparoscopy: a case report. AB - An unusual case of Richter's hernia on the 5th day after laparoscopy is presented. The complaints simulated a postoperative hematoma, but they became so intense that the expansion was surgically explored: a conservative procedure on the loop of small bowel was possible. To prevent this complication after laparoscopy careful shaking of the abdominal wall at removal of the instruments is necessary. PMID- 2968929 TI - Surface antigens of theophylline-resistant and theophylline-sensitive human E rosette-forming T lymphocytes. AB - Theophylline-resistant (TER) and theophylline-sensitive (TES) subsets of E rosette-forming T cells were analysed for surface antigens detected by monoclonal antibodies Leu-2, Leu-15, Leu-3, and Leu-8. Two-colour analysis was performed by laser flow cytometry. About 30% of TER cells was classified as Leu-2+, 15- precursor cytotoxic T lymphocytes and about 50% as 3+,8+ (T inducers). TES fraction was deficient in cytotoxic 2+,15- cells and contained lymphocytes belonging to suppressor circuit. Leu-3+,8- (helpers and suppressor-amplifiers) were present in both fractions. PMID- 2968931 TI - Phosphorylation of myosin in smooth muscle and non-muscle cells. In vitro and in vivo effects. PMID- 2968932 TI - Decreased acylation of phosphatidylcholine in diabetic rat erythrocytes. AB - We investigated the effects of streptozocin-induced diabetes on composition and metabolism of rat erythrocyte lipids. Diabetes produced no change in contents of cholesterol, total phospholipids, and proportions of phosphatidylcholine and phosphatidylethanolamine in phospholipids. The acylation of total phospholipids with palmitic, oleic, or arachidonic acids was decreased (P less than .01) in intact erythrocytes from diabetic versus control animals. This anomaly was underlaid by a decrease (P less than .01) in acylation of phosphatidylcholine, whereas phosphatidylethanolamine was unaffected. The impaired acylation of phosphatidylcholine was unchanged in vitro by insulin or coenzyme A but was restored to control values by ATP and by insulin treatment of the diabetic rats. We conclude that diabetes specifically alters the acylation of at least phosphatidylcholine in rat erythrocyte, an effect that might modify the remodeling of erythrocyte phospholipids and thereby the membrane function. PMID- 2968933 TI - Persistent grossly elevated plasma immunoglobulin A levels in untreated streptozocin-induced diabetic rats. AB - Experimental diabetes mellitus was induced in adult male and female rats by injecting streptozocin (STZ; 60 mg/kg i.p.) in preparation for a screening survey of changes in the pattern of undenatured plasma proteins, as revealed by two dimensional (2-D) gel electrophoresis followed by silver staining. As early as 8 12 days later, the 2-D gels revealed three high-molecular-weight plasma protein spots, which persisted for 150 days in the blood of untreated diabetic rats. Such spots were not seen in plasma of normal control rats. Evidence is presented for the presumptive characterization of these proteins as oligomers of immunoglobulin A (IgA). Specific measurement of total IgA content of diabetic plasma samples by single-radial immunodiffusion, after reduction with dithiothreitol and alkylation with iodoacetamide, reveals that IgA content increases linearly from control values of 11.1 +/- 4.6 to 358 +/- 249 mg/dl (means +/- SE) 21 days after STZ and persists at these high levels for as long as 150 days. Diabetic rats injected daily with insulin showed IgA levels only two to four times higher than normal. Neither experiments designed to quantitate the rates of clearance (catabolism plus excretion) of 125I-labeled secretory IgA from the circulation of normal and diabetic rats nor measurement of total IgA in the bile from diabetic and normal bile fistula rats supports the view that slowed clearance from the circulation or impaired biliary excretion in the diabetic rat causes observed gross hyperimmunoglobulinemia A. PMID- 2968934 TI - Increased plasma IgA, sIgA, and C3- and IgA-containing immune complexes with renal glomerular deposits in diabetic rats. AB - Male Sprague-Dawley rats were fasted 18 h and given streptozocin (STZ; 60 mg/kg body wt i.p.). The resultant diabetes mellitus, not treated with insulin, was associated with persistent manifoldly increased plasma IgA levels, as measured by single-radial immunodiffusion after reduction with dithiothreitol and alkylation with iodoacetamide. Also observed were concurrent increases in plasma levels of secretory IgA (sIgA) and of C3- and IgA-containing immune complexes (C3-IgA-CIC). After 104 days without insulin treatment, six of the diabetic rats were given daily injections of 2 U of insulin for 11 days. Insulin treatment was associated with a precipitous decrease in plasma levels of IgA, sIgA, and C3-IgA-CIC. Cessation of insulin treatment resulted in restoration of greatly increased levels of all three IgA-containing species. Histoimmunofluorescence studies of kidneys from untreated rats with diabetes of 192-324 days revealed glomerular capillary wall and mesangial deposits reacting strongly with anti-IgA (alpha chain-specific) antiserum. Kidneys from two of the diabetic rats (324 days) were tested with anti-rat C3 and anti-rat secretory component (SC) antisera, and they reacted positively. Control kidneys from normal rats examined simultaneously were negative. The concurrent changes in plasma levels of three IgA-containing species in the untreated STZ-induced diabetic rat and the demonstration of abnormal immunoreactive IgA-containing renal glomerular deposits make this experiment an attractive model for studying the possible role of disturbed IgA metabolism in the pathogenesis of diabetic nephropathy. PMID- 2968935 TI - Mechanisms of nitrosourea-induced beta-cell damage. Activation of poly (ADP ribose) synthetase and cellular distribution. AB - It has been hypothesized that the critical step in streptozocin (STZ)-induced beta-cell toxicity is the overactivation of the nuclear enzyme poly(ADP-ribose) synthetase resulting from DNA strand breaks. Overactivation of this enzyme leads to a lethal depletion of its substrate, NAD, in the beta-cell. However, recently it has been shown that a lethal concentration of STZ and a nontoxic concentration of its nitrosoamide moiety methylnitrosourea (MNU) damage beta-cell DNA to the same extent and cause comparable amounts of DNA strand breaks. This study was performed to determine whether STZ and MNU activate poly(ADP-ribose) synthetase to the same extent. Monolayer cultures of islet cells from neonatal rats were exposed to concentrations of MNU and STZ of 10(-3) to 10(-2) M. The results show that both chemicals caused comparable activation of the enzyme at all concentrations tested. These data demonstrate that activation of poly(ADP-ribose) synthetase alone is not the critical step in STZ-induced beta-cell toxicity. Based on this finding, it appeared possible that STZ may be selectively sequestered into some critical site in the beta-cell other than the nucleus. Therefore, studies were initiated with 14C-labeled STZ and MNU to determine whether STZ might be distributed in the beta-cell differently than MNU. Total cellular DNA and protein from both RINr (clone 38) and islet cell monolayers were separated on hydroxylapatite columns after exposure to 14C-labeled chemicals. The amount of label incorporated into each fraction was determined by liquid scintillation spectrometry, and the ratio of label incorporated in protein to that in DNA was determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968937 TI - Observations on offspring of mice made diabetic with streptozocin. AB - My objective was to determine the effect of streptozocin (STZ)-induced diabetes in male inbred mice on nonfasting blood glucose levels and body weights of offspring. Hyperglycemia was induced in CBA/H male mice by either multiple subdiabetogenic doses (MD) of STZ (5 doses/day of 50 mg STZ/kg body wt) or by a single high sublethal (SD) dose (200 mg STZ/kg body wt). Females were made diabetic by the multiple low-dose procedure. The following matings were set up: SD males with normal (NOR) females; MD males with NOR females; NOR males with MD females; MD males with MD females. Controls were matings of NOR males with NOR females. Among the first cohort of litters was born one female from the cross of an SD male with an NOR female who became spontaneously hyperglycemic at 5 wk of age; the female progeny of this cross had significantly lower body weights. All other progeny groups were normoglycemic (up to 5 wk) and had normal body weights. Test progeny weaned in the second and subsequent cohorts of litters were also normoglycemic. The major effect in this progeny group was on body weight; diabetic fathers (particularly MD males) mated with NOR females produced offspring with significantly higher juvenile body weights than the controls (increase of approximately 0.5 g). These body-weight distributions also appeared more homogeneous than the more variable body-weight distribution of the controls. In contrast, MD mothers (mated with NOR males) produced second-cohort offspring with significantly lower average body weights (decrease of approximately 1-2 g) than the controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968940 TI - The anabiosis of laparoscopy? PMID- 2968936 TI - Mechanisms of nicotinamide and thymidine protection from alloxan and streptozocin toxicity. AB - A common mechanism has been proposed for the beta-cell toxins alloxan (ALX) and streptozocin (STZ) involving the formation of single-strand breaks in DNA that lead to the overactivation of the enzyme poly(ADP-ribose) synthetase and the critical depletion of its substrate NAD. If the toxins act via this common mechanism, the poly(ADP-ribose) synthetase inhibitors nicotinamide and thymidine would be expected to affect the formation of DNA single-strand breaks in a similar fashion. To test the effects of these inhibitors, the formation of single strand breaks in the DNA of insulin-secreting RINr cells was monitored by assessing changes in the supercoiling of nucleoids after exposure to STZ, ALX, or methylnitrosourea (MNU). With the inclusion of nicotinamide or thymidine and STZ or MNU, more single-strand breaks in RINr cell DNA were detected. These results would be expected if nicotinamide and thymidine acted through inhibition of poly(ADP-ribose) synthetase. However, when the inhibitors were used in combination with ALX, fewer single-strand breaks were present. This suggests a reduction in ALX-induced hydroxyl radicals available to interact with DNA. Because nicotinamide has been demonstrated to be a hydroxyl-radical scavenger, the ability of thymidine to scavenge hydroxyl radicals was investigated. Thymidine, like nicotinamide, was found to be a potent scavenger of hydroxyl radicals. Thus, the mechanisms by which nicotinamide and thymidine protect against the toxic effects of STZ or ALX appear different. These findings suggest that the actions of beta-cell toxins are more complex than simply the overactivation of a single enzyme. PMID- 2968938 TI - Gastric mucosal repair in the cat: role of the hyperemic response to mucosal damage. AB - This study concerns the significance for the mucosal repair process of the gastric mucosal hyperemic response after mucosal damage caused by 2 M NaCl. Celiac artery blood flow was measured by Doppler ultrasound and stomach arterial inflow was either left undisturbed or reduced in a controllable fashion by tightening a vessel loop around the celiac artery immediately after mucosal exposure to 2 M NaCl or 150 mM NaCl. The stomach lumen was perfused with saline at pH 1.00 before and after exposure to 2 M NaCl. Gastric mucosal blood flow increased after exposure to 2 M NaCl, and 90 min after mucosal exposure to 2 M NaCl the mucosal surface showed nearly complete restitution of the surface epithelium. In all animals in which mucosal hyperemia was restricted by reducing celiac artery blood flow by 60% after mucosal damage, extensive gastric erosions were present. Similar reduction of celiac artery blood flow in control animals without mucosal damage did not produce visible lesions, and the stomachs proved to be normal by microscopy. These findings show that the gastric mucosal hyperemic response after damage is important for protection of the damaged mucosa during restitution of the gastric surface epithelium. PMID- 2968939 TI - The use of laparoscopy in retroperitoneal pathology. AB - Laparoscopy is usually not performed for retroperitoneal disease. However, if the retroperitoneal process is palpable or displaces viscera, laparoscopy may allow visualization and directed biopsy. In a personal series of 316 laparoscopies reviewed retrospectively, 19 (6%) were performed primarily for retroperitoneal pathology. All patients had CT scans documenting a retroperitoneal process. A confirmed histologic diagnosis was obtained on directed laparoscopic biopsy in 16 patients, including 9 non-Hodgkin's lymphomas. There was no mortality or morbidity in this series. Laparoscopy is a useful modality in selected patients with retroperitoneal diseases, and its use should be considered when a histologic diagnosis is necessary. PMID- 2968941 TI - Interaction of flavonoid compounds with contractile proteins of skeletal muscle. AB - Flavonoids (quercetin, rutin) influence ATPase activity and actomyosin superprecipitation. Low concentrations (below 20 mumol/l) of flavonoids were found to cause conformational changes in the myosin structure accompanied by an increase in ATPase activity. At higher concentrations an inhibitory action of flavonoids on both ATPase activity and actomyosin superprecipitation occurred. Conformational changes are likely to be due to flavonoids binding to regulatory site near the active centre of the myosin head. The effect of quercetin was stronger than that of rutin. PMID- 2968942 TI - The treatment of advanced ovarian carcinoma (II): interval reassessment operations during chemotherapy. PMID- 2968944 TI - [Hepatitis B vaccine]. PMID- 2968943 TI - Interexaminer variability of fetal Doppler velocity waveforms. AB - A study was made of the interexaminer variability of Doppler velocity waveform determinations of the fetal circulation. Three investigators recorded Doppler ultrasound tracings of the blood velocity in the fetal thoracic and abdominal aorta and umbilical artery in 23 uncomplicated pregnancies. Of these recordings, a hard copy was made, and the pulsatility indices were computed. An analysis of variance was performed and the reliability calculated. The interexaminer variability was nonsignificant in measurements of the three blood vessels under examination. Both thoracic and abdominal aorta measurements showed poor reliability (intraclass correlation coefficient of reliability 0.30 and 0.19, respectively) due to high random errors. Umbilical artery measurements showed a better reliability (intraclass correlation coefficient of reliability 0.61), and, therefore, these hold the greatest clinical applicability. PMID- 2968945 TI - Prevalence of isolated antibody to hepatitis B core antigen in an area endemic for hepatitis B virus infection: implications in hepatitis B vaccination programs. AB - Of 1,801 Chinese subjects, age 1 to 90 years, screened for hepatitis B surface antigen and antibody (HBsAg, anti-HBs) and antibody to hepatitis B core antigen (anti-HBc), 214 (11.9%) had an isolated, positive anti-HBc result; anti-HBc was reproducibly present in the initial sera in only 66% and persisted after an interval of 2 weeks to 3 months in only 73%. There was a strong correlation between the rates of reproducibility and persistence of isolated anti-HBc and the initial anti-HBc titers. Thirty-two subjects with persistent, isolated anti-HBc received four doses of hepatitis B vaccine (5 micrograms, HEVAC B) at 0, 1, 2 and 12 months: 56% developed a primary anti-HBs response in response to hepatitis B vaccine, 16% developed an anamnestic or secondary anti-HBs response, and 28% were undetectable for anti-HBs even after four doses of vaccine. The low rates of reproducibility and persistence of anti-HBc together with the high rate of primary anti-HBs response to hepatitis B vaccine in subjects with isolated anti HBc raise doubts as to the reliability of anti-HBc (Corzyme, Abbott Laboratories, North Chicago, Ill.) as a single screening test for hepatitis B infection prior to vaccination and suggests that subjects with isolated anti-HBc, in particular those with low anti-HBc titers, be included in vaccination programs. PMID- 2968947 TI - Control of hepatitis B virus among Alaskan natives. PMID- 2968946 TI - Thallium-201 per rectum for the diagnosis of cirrhosis in patients with asymptomatic chronic hepatitis. AB - In normal subjects, thallium-201, administered per rectum, is taken up mainly by the liver (heart/liver ratio in normal subjects: 0.04 to 0.12). It has been claimed that an increased heart/liver ratio is suggestive of portal-caval shunting and portal hypertension. To evaluate the possibility of using thallium 201 as a test to diagnose cirrhosis, we administered this substance per rectum to 33 patients with biochemical evidence, but no clinical symptoms, of liver disease. Laparoscopy and liver biopsy revealed chronic active hepatitis without cirrhosis in 18 patients, and chronic active hepatitis with cirrhosis in the others. The results of conventional liver function tests were similar in both groups. A significant difference, however, was found between the means of fasting serum bile acid concentrations (9.8 +/- 3.2 and 18.3 +/- 4.2 microM per liter) in chronic active hepatitis without cirrhosis and cirrhotic patients, and between the means of the heart/liver ratios 20 min after thallium-201 administration (heart/liver: 0.09 +/- 0.03 and 0.54 +/- 0.13, respectively). Unlike the serum bile acid concentration which gave some overlapping values, the thallium-201 test clearly distinguished the chronic active hepatitis without cirrhosis group from the cirrhotics. In the cirrhotic group, there was a significant correlation between the heart/liver ratio and signs of portal hypertension such as esophageal varices, increased diameter of the vena porta and hypersplenism. The thallium-201 test is therefore useful in discriminating between chronic active hepatitis with and without cirrhosis in clinically asymptomatic subjects with biochemical evidence of moderate liver function impairment. A heart/liver uptake ratio much higher than normal (above 0.30) strongly suggests the development of hepatic cirrhosis. PMID- 2968949 TI - Human factor H (beta 1H-globulin): linkage analysis. AB - Linkage data on human factor H (HF) and 22 other human genetic markers are presented. Close linkage at theta less than 0.10 can be ruled out for a series of marker systems (Rh, PGM1, ACP1, Jk, Tf, Gc, MNSs, ME2, HLA, GLO1, ORM, Gt, PI, Hp, GPT). Strong evidence for linkage was obtained for peptidase A (PEPA) with lods greater than 3.0 at theta = 0.10 in males and at theta = 0.20 for the sexes combined. From this result the HF locus can be provisionally assigned to chromosome 18. PMID- 2968948 TI - Genotypic and phenotypic characteristics of multiple myeloma. PMID- 2968951 TI - Non-specific regulatory mechanism of contact sensitivity: the requirement of intermediate cells for non-specific suppressor factor (NSF) activity. AB - Non-specific suppressor factor (NSF), which inhibits passive transfer of contact sensitivity (CS), is produced spontaneously from macrophage-like suppressor cells which were induced by intravenous (i.v.) administration of oxazolone (Ox) conjugated spleen cells. NSF is absorbed with normal spleen cells, and NSF treated spleen cells acquire the ability to suppress the transfer of the effector cell function of CS non-specifically. In the present study, the events involved in the suppression by NSF were investigated. The involvement of intermediate cells between NSF and effector T cells in the suppression by NSF was suggested by the following observations: (i) NSF was absorbed with plastic-adherent and cyclophosphamide (CY)-sensitive non-T cells present in normal spleen cells; (ii) deletion of plastic adherent and CY-sensitive cells but not of adult thymectomy (ATx)-sensitive cells from the effector cell population, rendered the effector cells resistant to the suppressor activity of NSF; (iii) reconstitution of CY pretreated effector cell population with Thy-1-negative spleen cells restored the ability of NSF to suppress CY-pretreated effector cells function. On the contrary, reconstitution with Ia-negative spleen cells did not restore the ability of NSF to suppress CY-pretreated effector cells function. Thus, NSF may not suppress directly the effector T-cell function, but intermediate cells, which are possibly macrophage-like cells, may exert a suppressive role after absorbing NSF. Species specificity was observed between the interaction of NSF and intermediate cells. The possible role of the intermediate cells in the suppression circuit of CS by NSF is discussed. PMID- 2968952 TI - Atrial natriuretic polypeptide in bovine adrenal medulla. AB - Two radioimmunoassays for alpha-human atrial natriuretic polypeptide (alpha-hANP) with different specificities were used to study the tissue level and the nature of alpha-hANP-like immunoreactivity in the bovine adrenal gland. A considerable amount of alpha-hANP-like immunoreactivity was detected in the adrenal medulla (90.8 +/- 21.1 and 90.0 +/- 23.1 ng/g with the two radioimmunoassays), while no detectable amount (less than 1.0 ng/g) was present in the cortex. Gel permeation chromatographic analysis showed that ANP in the medulla is composed of two components of alpha-hANP-like immunoreactivity with high and low molecular weights in the approximate ratio of 2:1, eluting at the elution positions of gamma-hANP and alpha-hANP, respectively. Reverse-phase high performance liquid chromatographic analysis revealed that alpha-hANP-like immunoreactivity with a low molecular weight in the medulla consists of two major components, which comigrate with synthetic alpha-hANP(5-28) and alpha-hANP. When cultured bovine adrenal chromaffin cells were incubated in the presence of nicotine (10(-5) M), alpha-hANP-like immunoreactivity was released into the medium concomitantly with catecholamines from chromaffin cells. These findings indicate that a discrete ANP system is present in the adrenal medulla and that ANP is cosecreted with catecholamines from chromaffin cells, suggesting the possible involvement of ANP in the adrenomedullary function. PMID- 2968950 TI - Immune-complex inhibition of macrophage activation is not due to an interaction with the binding or processing of IFN-gamma. AB - Preincubation of macrophages with immune complexes suppresses the interferon gamma (IFN-gamma) induction of tumouricidal activity and Ia surface expression. The studies reported in this manuscript were designed to test if immune complexes alter the interaction of IFN-gamma with macrophages. The binding of IFN-gamma to its specific cellular receptor, the uptake or the degradation were not affected by preincubation of macrophages with immune complexes. Preincubation of macrophages with high doses of phorbol esters mimics the inhibitory effect of immune complexes. This suggests that a strong activation of protein kinase C suppresses a subsequent activation of macrophages by IFN-gamma. PMID- 2968953 TI - Effects of intravenously administered beta-human atrial natriuretic polypeptide in humans. AB - beta-Human atrial natriuretic polypeptide (beta-hANP) is an antiparallel dimer of alpha-human ANP (alpha-hANP) that was isolated from human atria. Using synthetic beta-hANP and a radioimmunoassay for alpha-hANP that also detects beta-hANP, we have previously demonstrated that beta-hANP is converted into alpha-hANP in human plasma in vitro. In the present study, we compared the effects of intravenous administration of beta-hANP (100 micrograms) to five normal human volunteers with those of an equimolar administration of alpha-hANP (50 micrograms) to the same subjects, and we also investigated the possible mechanisms of actions of beta hANP. Although the administration of alpha-hANP caused a significant decrease in blood pressure with a reactional increase of heart rate, beta-hANP elicited minimal change of blood pressure. In contrast, beta-hANP exerted more potent and longer lasting diuretic and natriuretic activities than did alpha-hANP. Net changes in urine volume and sodium excretion induced by beta-hANP (579 +/- 65 ml, 56.0 +/- 9.9 mEq) were significantly greater than those elicited by alpha-hANP (396 +/- 50 ml, 34.7 +/- 4.9 mEq; p less than 0.05, respectively). The administration of beta-hANP revealed a longer retention of the ANP-like immunoreactivity level in plasma, compared with that of alpha-hANP. High performance gel permeation chromatography coupled with the radioimmunoassay revealed that beta-hANP (Mr = 6000) was also converted into alpha-hANP (Mr = 3000) in human plasma in vivo. The demonstrated conversion of beta-hANP into alpha-hANP could be relevant to the observed effects of beta-hANP in humans. PMID- 2968954 TI - Impaired release of atrial natriuretic factor in NaCl-loaded spontaneously hypertensive rats. AB - Our previous studies demonstrated that NaCl-sensitive spontaneously hypertensive rats (SHR) of the Okamoto strain exhibit increased blood pressure and reduced noradrenergic input to the anterior hypothalamus area when fed high NaCl diets. The current study tested the hypotheses that 1) release of atrial natriuretic factor (ANF) into the plasma is impaired in NaCl-loaded SHR, a defect that would tend to elevate blood pressure, and 2) ANF levels in regions of brain involved in blood pressure regulation, such as the anterior hypothalamic area, are altered in SHR. SHR and control Wistar-Kyoto rats (WKY) were placed on 1% or 8% NaCl diets at age 7 weeks; 2 weeks later, ANF levels were measured in plasma, left and right atria, anterior hypothalamic area, ventral hypothalamic area, posterior hypothalamic area, pons, and medulla by radioimmunoassay. Blood for ANF assay was obtained from intra-arterial cannulas in conscious, unrestrained rats studied in the resting state. The 8% NaCl diet produced an increase in blood pressure in the SHR, but not in the WKY. Plasma ANF levels were significantly greater in WKY fed 8% NaCl than in WKY fed 1% NaCl, but dietary NaCl loading did not produce similar increases in plasma ANF in the SHR. Plasma ANF levels were not significantly different between SHR and WKY fed the 1% NaCl diet. The observation that dietary NaCl loading stimulated ANF release into the plasma in WKY but not in SHR suggests that the exacerbation in hypertension seen in NaCl-loaded SHR may be related to an impairment in ANF release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2968955 TI - Genetics and salt modulate renal responses to atrial natriuretic factor. AB - We examined the consequences of genetic susceptibility or resistance to NaCl induced hypertension and of prior salt loading (high or low NaCl intake) on the responses of isolated perfused Dahl salt-sensitive (DS) and Dahl salt-resistant rat (DR) kidneys to atriopeptin II. Atriopeptin II increased the glomerular filtration rate only in kidneys from high NaCl-fed rats, irrespective of their DS or DR status. Superimposition of norepinephrine on atriopeptin II further increased the glomerular filtration rate only in kidneys from low NaCl-fed rats (which had not reacted to atriopeptin II alone), irrespective of their DS or DR status, and did not change the glomerular filtration rate of high NaCl-fed rats. Norepinephrine alone, without atriopeptin II, uniformly decreased the glomerular filtration rate by about 80%. Atriopeptin II increased sodium excretion of high NaCl and low NaCl DR kidneys by more than five times as much as in the corresponding DS kidneys. Therefore, the glomerular filtration rate response to atriopeptin II varied globally with dietary NaCl, independently of genetic predisposition or resistance to NaCl-induced hypertension. The natriuretic response to atriopeptin II was blunted in kidneys from rats genetically susceptible to NaCl-induced hypertension, independently of their NaCl consumption. Atriopeptin II also ameliorated or reversed the adverse effect of norepinephrine on the glomerular filtration rate. PMID- 2968958 TI - Induction of tumor suppression and delayed-type footpad reaction by transfer of lymphocytes sensitized to a xenogenized tumor variant. AB - Splenocytes immune to a highly immunogenic ("xenogenized", L5178Y/DTIC) variant of a murine lymphoma exert anti-parental-tumor activity in a systemic adoptive transfer system, the effect being apparently associated with the Lyt-2- fraction of the lymphocyte population. During investigation of the mechanisms of this protection, we found that the L5178Y/DTIC tumor-immune lymphocytes exhibited an appreciable anti-L5178Y delayed-type hypersensitivity (DTH) response. Enrichment of those lymphocytes in L3T4+ cells significantly enhanced the protective effect as well as the DTH reaction, whereas the use of an anti-L3T4 but not anti-Lyt-2 reagent blocked both activities. In vitro, lymphocyte proliferation against L5178Y cells occurred and was apparently associated with the Lyt-2- fraction of a population of L5178Y/DTIC immune splenocytes. PMID- 2968957 TI - Con A-propagated, auto-reactive T cell clones that secrete factors promoting high IgA responses. AB - Four BALB/c T cell clones from among a set propagated in the presence of concanavalin A (Con A) were selected on the basis of their ability to produce supernatant factors promoting high IgA plaque-forming cell (PFC) responses by 2,4,6-trinitrophenyl-conjugated keyhole limpet hemocyanin (TNP-KLH)-primed splenic B cells in the presence of TNP-SRBC. Such clones could be derived from cultures containing T cells not only from gut-associated lymphoid tissue, but also from the spleen. The selected clones all proliferated well in the presence of syngeneic, irradiated APC without either Con A or exogenous IL-2, but required both APC and Con A to produce helper factors. Factors from three of the clones helped B cells both to proliferate and to differentiate into IgM, IgG and IgA PFC. Factors from the fourth clone helped B cells differentiate into IgA and IgG PFC and may have promoted switching to these isotypes but did not support either B cell proliferation or generation of IgM PFC. Cross-linking of B cell receptors for antigen was not required for the response to the helper factors since TNP SRBC were unnecessary and high concentrations of them were actually inhibitory. PMID- 2968959 TI - Hereditary palmoplantar keratoderma and dermatophytosis. PMID- 2968956 TI - Regulation of human IgE synthesis by soluble factors. Papain treatment of a FcE receptor-positive B-cell line (RPMI-1788) releases regulatory factors for IgE synthesis. AB - A novel mechanism for the release of helper and suppressor factors for human IgE synthesis is described. When FcE receptor-positive RPMI-1788 cells are treated with papain, a helper factor(s) for human IgE synthesis is released. At the same time a significant decrease in the number of cell surface FcE receptors is observed. The immunoglobulin synthesis-enhancing activity is IgE isotype-specific inasmuch as the same supernatant suppresses the synthesis of human IgA myeloma cells. When the FcE receptor-positive RPMI-1788 cells are treated with tunicamycin and then with papain, a suppressor factor(s) for human IgE synthesis is released. The mechanism by which these factors affect human myeloma IgE synthesis is unclear at present. Our results indicate that enhanced IgE synthesis is not due to increased numbers of secreting cells nor to an increased release of presynthesized IgE. In summary, papain treatment of FcE receptor-positive, but not FcE receptor-negative cells, generates a factor that regulates IgE synthesis. These results also provide evidence for the close relationship between the IgE regulatory factors and the low affinity receptors for IgE present on lymphocytes. PMID- 2968960 TI - Sezary cell count in exfoliative dermatitis. AB - Sezary cells were present in a number of less than 1000/mm3 in 52 of 78 cases of various benign diffuse dermatoses and mycosis fungoides. The count was higher than 1000/mm3 in two cases of Sezary syndrome in which the histopathology was that of mycosis fungoides and in six cases of exfoliative erythroderma of undetermined etiology with the histopathology of nonspecific chronic dermatitis. One of these six cases developed palmoplantar keratoderma, but abnormal cells were not present in the tissue. The diagnosis of Sezary syndrome in these cases of exfoliative erythroderma could not be established, even though the Sezary cell count was more than 1000/mm3, because the abnormal cells were absent in the skin biopsy specimen. It is concluded that the number of Sezary cells present is not significant for the diagnosis of Sezary syndrome. Our cases with the diagnosis of Sezary syndrome are an erythrodermic variant of mycosis fungoides rather than a progressive conversion from exfoliative erythroderma. PMID- 2968961 TI - Ketoconazole-induced fixed drug eruption. PMID- 2968962 TI - Tinidazole-induced fixed drug eruption. PMID- 2968963 TI - Epidemiology of contact allergy in Singapore. AB - The prevalence of contact allergy in 2471 patients patch tested in Singapore was 49.2% (571/1160) in women and 49.8% (653/1311) in men (sex difference not significant). The rate appeared to increase with age (43.7% in those less than 20 years, 50.6% between 20 and 49 years, and 63.1% in those greater than 49 years). There was no significant difference in the prevalence of contact allergy among the major races (Chinese, Malay, and Indian) of Singapore. Contact allergies in 16.8% (96/571) of women and 38.9% (254/653) of men (p less than 0.001) were occupational. Nickel, fragrance mix, proflavine, and chromate were common sensitizers. The male:female prevalence of nickel allergy was 1:2 (sex difference p less than 0.001) and for chromate was 7:1 (sex difference p less than 0.001). Chromate sensitivity was occupational in 87.7% (121/138) of men and 47.1% (8/17) of women (sex difference p less than 0.001). Proflavine, neomycin, and clioquinol were the more common medicament sensitizers. PMID- 2968964 TI - Tryptophan reduction and histidine racemization during deprotection by catalytic transfer hydrogenation of an analog of the luteinizing hormone releasing factor. AB - (D-Trp)6-LHRH:pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-GlyNH2 was prepared by solid phase peptide synthesis using the nitro group to protect the guanidine side chain of the arginyl residue. Removal of the side-chain protecting groups was carried out by catalytic transfer hydrogenation (CTH) using palladium acetate/ammonium formate or palladium on charcoal/formic acid. We show in this paper that this deprotection method induces i) reduction of the tryptophan residue and ii) epimerization at the histidine level (with palladium acetate/ammonium formate). Despite the formation of significant amounts of reduced peptide, CTH enabled us to obtain (D-Trp)6-LHRH in relatively good yield. PMID- 2968966 TI - Immune suppression gene on HLA-Bw54-DR4-DRw53 haplotype controls nonresponsiveness in humans to hepatitis B surface antigen via CD8+ suppressor T cells. AB - The development of antiviral vaccines has been accelerated using monoclonal antibody and/or recombinant DNA techniques, the objective being to prevent grave viral infectious diseases, such as acquired immunodeficiency syndrome (AIDS), adult T-cell leukemia (ATL), and hepatitis B virus (HBV)-associated liver diseases. Certain proportions of individuals in the human population do not have any appreciable immune response to foreign antigens, either in cases of natural exposure or a planned immunization. Here we report that in the nonresponders to HB vaccine, there is an HLA-linked immune suppression gene for hepatitis B surface antigen (Is-HBsAg) controlling the nonresponsiveness to HBsAg through HBsAg-specific suppressor T cells. The Is-HBsAg is in strong linkage disequilibrium with the HLA-Bw54-DR4-DRw53 haplotype. PMID- 2968965 TI - Monoclonal antibody 4B4 blocks suppression of the immune response to Cryptomeria japonica pollen antigen. AB - The monoclonal antibody 4B4 blocked the T8+ T cell-mediated suppression of IgE response to Cryptomeria japonica pollen antigen (CPAg). The helper T cells for IgE response to CPAg expressed 4B4 specificity, whereas the suppressor T cells did not. The 4B4 molecule on PBL was a heterodimer and three additional bands were detected on activated CPAg-specific T4+ T cells. They were not closely associated with HLA molecules. It is thus conceivable that the 4B4 molecule is not only a marker of helper T cells but also has an important role in the suppressor-helper interactions in the regulation of IgE response to CPAg. PMID- 2968967 TI - Late onset of renal dysfunction in survivors of bone marrow transplantation. AB - Between 1980 and 1986, 44 children with acute lymphoblastic leukemia (ALL) or Stage IV neuroblastoma (NB) underwent allogeneic or autologous bone marrow transplantation (BMT). Twenty-nine of these patients were alive and in remission 3 months post BMT and were evaluable for this analysis of whom eleven have developed renal dysfunction. Six of 17 (35%) evaluable ALL patients developed renal dysfunction (3.5 to 6 months post BMT). This group was transplanted for CALLA positive ALL and received an autologous transplant. Preparation included tenopiside (VM 26) cytosine arabinoside, and cyclophosphamide followed by total body irradiation (TBI). One patient received 850 cGy in a single fraction, while all other patients received fractionated TBI (1200-1400 cGy in 6-8 fractions over 3-4 days). Five of 7 (71%) evaluable patients who received a BMT for NB have developed late renal problems (4-7 months after BMT). The preparation for NB patients included VM 26, cis-platinum, melphalan, cyclophosphamide, and fractionated TBI (1200-1296 cGy). All seven NB patients had received cis-platinum as induction treatment prior to transplantation. All patients presented with anemia, hematuria, and elevations of BUN and creatinine. Two patients underwent renal biopsies which were consistent with radiation nephropathy or hemolytic uremic syndrome. In conclusion, a high incidence of renal dysfunction has occurred 3 to 7 months after BMT for children with NB and ALL. The clinical and laboratory features are consistent with either acute radiation nephropathy or hemolytic-uremic syndrome. These patients were prepared for BMT with multiple chemotherapeutic agents as well as TBI. The relatively young age of these patients and conditioning with intensive multi-agent chemotherapy may decrease the tolerance of the kidney to radiation injury. PMID- 2968968 TI - Comparative in-vitro activities of A-56268 (TE-031) and erythromycin against 306 clinical isolates. AB - The inhibitory (MIC) and bactericidal (MBC) activities of a new macrolide A-56268 (TE-031) against 306 clinical aerobic bacterial isolates was compared with that of erythromycin. The MIC90/MBC90 ratios for A-56268 were: Campylobacter jejuni 4/16, Haemophilus influenzae 8/8-16, H. parainfluenzae 8/8-16, Legionella pneumophila 0.06/0.5, methicillin-sensitive isolates of Staphylococcus aureus 0.5/1, and coagulase negative staphylococci 1/8, methicillin resistant isolates of Staph. aureus and coagulase negative staphylococci greater than 16/ greater than 16, Streptococcus pneumoniae 0.06/0.125, streptococcus Group A 0.06/2-4, streptococcus Group B 0.06/8- greater than 16, streptococcus Groups C and G 0.125/8 and Str. faecalis 4/64. Compared with erythromycin, A-56268 had greater inhibitory and bactericidal activity against isolates of L. pneumophila, with an MIC90 16-fold less and an MBC90 eight-fold less than that of erythromycin. Except for enterococci, A-56268 showed inhibitory activity equal to or greater than that of penicillin G against isolates of streptococci and an MIC two-fold less than that of erythromycin. For other strains tested, the inhibitory and bactericidal activities of A-56268 and erythromycin were similar. The clinical importance of the differences between these two macrolides will depend on the pharmacokinetic and tissue penetration properties of the new compound. PMID- 2968969 TI - The pharmacokinetics and extravascular diffusion of teicoplanin in rabbits and comparative efficacy with vancomycin in an experimental endocarditis model. AB - The serum protein binding, extravascular diffusion and urinary excretion of teicoplanin were studied in rabbits. Extravascular diffusion was studied after a 20 min iv infusion, and after one or five im injections (7.5 mg/kg), and was compared with the results obtained after im administration of vancomycin (7.5 and 15 mg/kg). In an experimental model of Staphylococcus aureus endocarditis, the efficacy of both antibiotics was also investigated. We observed teicoplanin serum protein binding of 87 +/- 4%. The serum concentrations of teicoplanin showed a three-phase exponential decline: T1/2 alpha, 0.11 +/- 0.01 h; T1/2 beta, 1.4 +/- 0.4 h; T1/2 gamma, 8.3 +/- 2.2 h. Teicoplanin appeared to be slightly secreted by renal tubules. The extravascular diffusion and the therapeutic efficacy of both drugs were studied with intervals between two injections based on the same multiple of beta half-life. Teicoplanin, like vancomycin, appeared slowly in extravascular fluid and the diffusibility of both drugs was similar. Peak extravascular concentrations of teicoplanin after 5 im injections were greater when the compound was administered every 16 h, rather than every 24 h and, for this drug, iv administration induced higher peak extravascular concentrations (P less than 0.01) than im injection. In the experimental model of S. aureus endocarditis, vancomycin 9 mg/kg/12 h and teicoplanin 4.5 mg/kg/16 h were similarly active and more effective than teicoplanin 4.5 mg/kg/24 h. PMID- 2968970 TI - Nonadrenergic bronchodilator mechanisms in normal human subjects in vivo. AB - In seven normal subjects we investigated whether a nonadrenergic bronchodilator nervous system is demonstrable in humans in vivo. After inhalation of leukotriene D4 (LTD4), respiratory resistance (Rrs) increased by 115 +/- 11% (SE). Subsequent inhalation of 2 nmol of capsaicin induced coughing and a fall in Rrs of 22.1 +/- 2% (P less than 0.01). However, inhalation of the diluent of capsaicin, 10% saline-ethanol, decreased Rrs similarly. These bronchodilator responses were not altered by inhaled ipratropium bromide (120 micrograms) and oral propranolol (80 mg). After ipratropium and propranolol, voluntary coughing alone decreased Rrs by 25 +/- 3% (P less than 0.05). We next investigated whether these bronchodilator responses could be blocked by anesthesia of the airways with inhaled lidocaine. After inhalation of lidocaine and LTD4, capsaicin aerosol induced coughing and a transient increase in Rrs of 18 +/- 6% (P less than 0.05) but no bronchodilation. Inhalation of saline-ethanol (n = 4) and a deep inhalation (n = 6) decreased Rrs by 18 +/- 4% (P less than 0.05) and 34 +/- 3% (P less than 0.001), respectively. We conclude that in normal subjects a nonadrenergic, noncholinergic bronchodilator mechanism exists, which can be activated by inhalation of capsaicin and inhibited by local anesthesia. PMID- 2968972 TI - The assessment and management of the failed back, Part I. AB - The 'failed back' represents a major clinical problem. Among the causes recognized for the 'failed back' is lateral root stenosis which has led to the use of electromyography (EMG) in the diagnosis of root compression. One hundred and thirty-two patients selected for surgical decompression were compared with 25 undergoing spinal fusion. EMG was found to be diagnostic in 95% of patients with bony canal stenosis, but only gave positive results in 62.5% of prolapsed disc patients. EMG improved in those patients who made a good recovery after surgery. PMID- 2968973 TI - The assessment and management of the failed back, Part II. AB - Despite adequate surgery a number of patients have a return of back pain and sciatica following operation, the so-called failed back. The results of a prospective study of 101 patients entering an intensive rehabilitation programme for the failed back is described. The programme consists of a team approach to the patient and his problems, using a variety of techniques to produce pain relief. Non-organic pain represents a significant problem in the failed back; 58% of patients were completely or substantially relieved of their pain. Transcutaneous nerve stimulation was found to be the single most useful treatment with exercise second. Non-organic pain was able to be relieved by this programme. PMID- 2968971 TI - Active glycolysis and glycogenolysis in early stages of primary cultured hepatocytes. Role of AMP and fructose 2,6-bisphosphate. AB - This study examines the factors involved in the rapid glycolysis and glycogenolysis that occur during the first stages of hepatocyte culture: a) Shortly after seeding glycolysis, estimated as lactate released to culture medium, increased 10 times in comparison to that reported in vivo. By 8 to 9 h of culture, hepatocytes were nearly glycogen-depleted even in the presence of insulin. b) 6-Phosphofructo-2-kinase remained 100% active during this period. The proportion of the initial active phosphorylase (87%) decreased to 57% by 7 h of culture. c) Fructose 2,6-bisphosphate content was initially similar to that found in liver of fed animals, decreased after seeding and increased thereafter up to four times the initial concentration. In spite of changes in the concentration of this activator, the glycolytic rate remained high and constant. d) ADP and AMP increased sharply after cell plating, reaching values 1.7 and 3.5 times higher. The rise in AMP levels may be involved in the activation of glycolysis and glycogenolysis, because this metabolite is known to act as an allosteric activator of phosphofructokinase and glycogen phosphorylase. This metabolic situation resembles that of cells under hypoxia. PMID- 2968974 TI - Psychological factors in the failed back. AB - Pain is a complex phenomenon. Lack of understanding of its nature leads to two common mistakes in the failed back, firstly to look for a discrete physical cause and secondly to ascribe pain to psychological causes. This paper explores the nature of pain in the failed back and discusses the implications for treatment. PMID- 2968975 TI - The failed back--an orthopaedic view. AB - The causes of the failed back syndrome are several. The failure to appropriately identify the structural sources of pain is one. The identities may include radiographically controlled injection procedures. The failure to use sufficiently discrete surgery with an attempt to avoid scar is another cause and the failure to provide an opportunity to avoid and later treat deconditioning is another. The appropriate treatment programme is based on objective assessment of function. Functional achievement as the principal care rather than pain self-report is the most effective manoeuvre to avoid the chronic pain of the failed back syndrome. PMID- 2968976 TI - The use of radiofrequency lesions for pain relief in failed back patients. AB - If conservative measures fail in the treatment of the failed back patient and if there is no indication for further surgery, interruption of nerve pathways conducting noxious stimuli may be attempted. The indication for such treatment is made on the result of a series of prognostic blocks analysing the conduction pattern of noxious stimuli. A new technique is described to interrupt the grey communicating ramus, conducting afferent fibres from the anterolateral and anterior parts of the annulus fibrosus. Results indicate a discrepancy between the result of radiofrequency lesions and the outcome of prognostic blocks. The discrepancy is more pronounced in failed back patients. Treatment with radiofrequency lesions is well tolerated and it has few adverse effects. It has a measure of success in a group of patients who are very difficult to manage otherwise. PMID- 2968977 TI - Liquid chromatographic determination of medroxyprogesterone acetate in tablets. AB - A reverse-phase liquid chromatographic method is described for the assay of medroxyprogesterone acetate in tablets. An octadecylsilane (C18) column with a mobile phase of methanol-0.01M dibasic ammonium phosphate (80 + 20 v/v, pH 7.2 +/ 0.1) and photometric detection at 254 nm separates medroxyprogesterone acetate from excipients. Detector responses were linear to concentrations of medroxyprogesterone acetate over the range 50-150 micrograms/mL (r = 0.999). Mean recovery of medroxyprogesterone acetate added to tablet excipients was 100.8%. Mean assay results were 101.3% (n = 3). The assay results are comparable to those obtained by the compendial liquid chromatographic method. PMID- 2968978 TI - Affinity labeling of the ATP-binding site of Ca2+-transporting ATPase of sarcoplasmic reticulum by adenosine triphosphopyridoxal: identification of the reactive lysyl residue. AB - Adenosine triphosphopyridoxal (AP3PL) was used as an affinity label directed toward the ATP binding site of the Ca2+-transporting ATPase of the rabbit skeletal muscle sarcoplasmic reticulum (SR). The reagent inhibited the ATPase activity competitively with ATP, Ki = 20 microM. Incubation of SR membranes with 100 microM AP3PL followed by treatment with NaBH4 resulted in 90% inactivation of the E-P forming activity as well as of the Ca2+-transporting activity. Adenosine di- and tetraphosphopyridoxals had similar but less pronounced effects on the Ca2+-transport system. AP3PL was bound to ATPase in a one-to-one stoichiometry in parallel with the loss of the enzymatic activities. ATP and ADP prevented the binding of AP3PL and thereby protected the enzyme from inactivation. The SR membranes were labeled with [3H]AP3PL and then digested with thermolysin in order to identify the attachment site of the affinity label. A 3H-labeled peptide (Val Glu-Pro-Ser-His-Lys* 684-Ser-Lys) was purified to homogeneity by Sephadex LH-20 chromatography and C18-reversed phase HPLC (Lys* denotes the binding site of [3H]AP3PL). These results indicate that the SR-ATPase peptide is folded in such a manner that Lys684 and Asp351, the phosphorylation site, are located very close to each other, since the distance between the 4-formyl group reacting with Lys684 and the gamma-phosphoryl group of the ATP moiety of AP3PL is rather small. PMID- 2968979 TI - A predicted tertiary structure of a thrombin inhibitor-trypsin complex explains the mechanisms of the selective inhibition of thrombin, factor Xa, plasmin, and trypsin. AB - The tertiary structure of a thrombin inhibitor-trypsin complex has been predicted by a molecular modelling considering the van der Waals interactions between the inhibitor and the enzyme. The selective inhibition of trypsin, thrombin, factor Xa, and plasmin exhibited by arginine and lysine derivatives has been clearly explained based on the predicted structure and the homology in the amino acid sequences of these enzymes. The differences in the amino acid sequences at the positions corresponding to Ile63, Leu99, and Ser190 of trypsin give each enzyme different binding affinities toward inhibitors and result in the selective inhibition. The X-ray analysis of the inhibitor-trypsin complex is in progress to prove the predicted structure. PMID- 2968980 TI - Biosynthesis of the mannose 6-phosphate recognition marker in transport-impaired mouse lymphoma cells. Demonstration of a two-step phosphorylation. AB - The biosynthesis of the mannose 6-phosphate recognition marker has been studied in transport-impaired mouse lymphoma cells to determine the subcellular location of the processing enzymes and to characterize the biosynthetic intermediates. Cells were labeled with [2-3H]mannose and chased at a low temperature (15 or 20 degrees C) or at 37 degrees C in the presence of m-chlorocarbonylcyanide phenylhydrazone to disrupt transport of the pulse-labeled molecules within the secretory apparatus. Both treatments inhibited the migration of the pulse-labeled glycoproteins to the Golgi apparatus as measured by the production of complex type asparagine-linked oligosaccharides. Despite this inhibition in protein transport, acid hydrolases were phosphorylated. Structural analysis of the phosphorylated oligosaccharides indicated that the transport-impaired cells produced a single species of phosphorylated high mannose oligosaccharide; essentially all of the molecules contain a single phosphodiester group that is restricted to the alpha 1,6 branch of the oligosaccharide. The results suggest that synthesis of mannose 6-phosphate-bearing high mannose oligosaccharides occurs in an ordered, compartmentalized posttranslational process. The initial phosphorylation of newly synthesized acid hydrolases occurs at a pre-Golgi site and results in the production of high mannose-type units that contain a single phosphodiester group. In a subsequent compartment, probably within the Golgi apparatus, the monophosphorylated units may be converted to diphosphorylated forms. Finally, at a site distal to the phosphorylation reactions the diesters are hydrolyzed to reveal the mannose 6-phosphate recognition marker. PMID- 2968981 TI - Associations of human erythrocyte band 4.2. Binding to ankyrin and to the cytoplasmic domain of band 3. AB - We have examined the associations of purified red cell band 4.2 with red cell membrane and membrane skeletal proteins using in vitro binding assays. Band 4.2 bound to the purified cytoplasmic domain of band 3 with a Kd between 2 and 8 X 10(-7) M. Binding was saturable and slow, requiring 2-4 h to reach equilibrium. This finding confirms previous work suggesting that the principal membrane binding site for band 4.2 lies within the 43-kDa cytoplasmic domain of band 3 (Korsgren, C., and Cohen, C. M. (1986) J. Biol. Chem. 261, 5536-5543). Band 4.2 also bound to purified ankyrin in solution with a Kd between 1 and 3.5 X 10(-7) M. As with the cytoplasmic domain of band 3, binding was saturable and required 4 5 h to reach equilibrium. Reconstitution with ankyrin of inside-out vesicles stripped of all peripheral proteins had no effect upon band 4.2 binding to membranes; similarly, reconstitution with band 4.2 had no effect upon ankyrin binding. This shows that ankyrin and band 4.2 bind to distinct loci within the 43 kDa band 3 cytoplasmic domain. Coincubation of ankyrin and band 4.2 in solution partially blocked the binding of both proteins to the membrane. Similarly, coincubation of bands 4.1 and 4.2 in solution partially blocked binding of both to membranes. In all cases, the data suggest the possibility that domains on each of these proteins responsible for low affinity membrane binding are principally affected. The data also provide evidence for an association of band 4.2 with band 4.1. Our results show that band 4.2 can form multiple associations with red cell membrane proteins and may therefore play an as yet unrecognized structural role on the membrane. PMID- 2968982 TI - Evidence of two steps in the homologous desensitization of vasopressin-sensitive phospholipase C in WRK1 cells. Uncoupling and loss of vasopressin receptors. AB - The exposure of WRK1 cells to arginine vasopressin (AVP), lysine vasopressin, or oxytocin for 18 h at 37 degrees C induced a homologous desensitization of the vasopressin- (VP) receptors. Dose-response curves of [3H]lysine vasopressin binding to control and desensitized WRK1 cells revealed a decrease in the maximal number of binding sites without any modification of its affinity (Kd values = 4.40 +/- 0.76 nM and 4.65 +/- 0.78 nM for control and desensitized conditions, respectively). The phenomenon was time- and dose-dependent. It was directly related to receptor occupancy, since the concentration of VP analogues leading to a half-maximal occupancy of VP receptors was closely related to the concentration of the corresponding analogue leading to a half-maximal decrease in VP-binding sites. It was also agonist-specific, since the V1 vasopressin antagonist desGly9d(CH2)5[D-Tyr(Et)2]VAVP was unable to affect the number of receptors. These desensitization processes were completely inhibited when the functional coated pits present in WRK1 cells were suppressed, indicating that the loss of VP binding sites was related to receptor internalization. The exposure of WRK1 cells to a vasopressin agonist for 18 h also led to an inhibition of the vasopressin sensitive phospholipase C activity. It was time- and agonist-dose-dependent, and occurred without any detectable changes in apparent affinity values (1.40 +/- 0.04 and 1.90 +/- 0.36 nM for control and desensitized cells, respectively). Control experiments showed that these inhibitions could not have been caused by a decrease in the labeling of inositol lipids. It is likely that they were mainly due to receptor internalization since (i) the hormonal treatment did not modify the basal level of phospholipase C; (ii) the maximal loss of VP-binding site was similar to the maximal inhibition of VP-stimulated IP accumulation; (iii) the recoveries of both VP-binding sites and VP-sensitive phospholipase C activity followed exactly the same time course (t1/2 = 4 h). In addition to this homologous desensitization of VP-sensitive phospholipase C activity, AVP also induced heterologous desensitization of bradykinin-sensitive phospholipase C activity. However, this effect was relatively weak (maximal inhibition 17 +/- 3%). The time course of VP-sensitive phospholipase C desensitization was more rapid than that of VP-receptors, indicating that desensitization involved at least two distinct steps, a rapid uncoupling step, and a later loss of vasopressin receptors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2968983 TI - Five distinct calcium and phospholipid binding proteins share homology with lipocortin I. AB - We have purified two 35-kDa proteins from rat peritoneal lavages that inhibit phospholipase A2 activity. Both are calcium/phospholipid-dependent membrane binding proteins and share similar structural and biochemical properties with lipocortins I and II. By sequence analysis we confirmed that they are lipocortin related, and we refer to the two inhibitors as lipocortins III and V. Using partial sequence information obtained from the purified rat proteins, full length cDNA clones for both proteins and for their human counterparts were isolated. As with lipocortins I and II, the amino acid sequences of lipocortins III and V which were deduced from the cDNA clones are highly conserved, sharing 50% identity with other family members. Related proteins were also purified from bovine intestinal mucosa and characterized by peptide mapping, sequence, and immunological analyses. In addition to lipocortins III and V the bovine preparation contained a third 35-kDa inhibitor and a 68-kDa inhibitor, extending the number of known lipocortins to six distinct proteins. While the various lipocortins are structurally similar, distinct differences in their cellular distribution indicate specialized roles for the individual proteins. PMID- 2968984 TI - Characterization of the stimulatory action of insulin on insulin-like growth factor II binding to rat adipose cells. Differences in the mechanism of insulin action on insulin-like growth factor II receptors and glucose transporters. AB - Insulin is known to increase the number of cell surface insulin-like growth factor II (IGF-II) receptors in isolated rat adipose cells through a subcellular redistribution mechanism similar to that for the glucose transporter. The effects of insulin on these two processes, therefore, have now been directly compared in the same cell preparations. 1) Insulin increases the steady state number of cell surface IGF-II receptors by 7-13-fold without affecting receptor affinity; however, insulin stimulates glucose transport activity by 25-40-fold. 2) The insulin concentration required for half-maximal stimulation of cell surface IGF II receptor number is approximately 30% lower than that for the stimulation of glucose transport activity. 3) The half-time for the achievement of insulin's maximal effect at 37 degrees C is much shorter for IGF-II receptor number (approximately 0.8 min) than for glucose transport activity (approximately 2.6 min). 4) Reversal of insulin's action at 37 degrees C occurs more rapidly for cell surface IGF-II receptors (t1/2 congruent to 2.9 min) than for glucose transport activity (t1/2 congruent to 4.9 min). 5) When the relative subcellular distribution of IGF-II receptors is examined in basal cells, less than 10% of the receptors are localized to the plasma membrane fraction indicating that most of the receptors, like glucose transporters, are localized to an intracellular compartment. However, in response to insulin, the number of plasma membrane IGF II receptors increases only approximately 1.4-fold while the number of glucose transporters increases approximately 4.5-fold. Thus, while the stimulatory actions of insulin on cell surface IGF-II receptors and glucose transport activity are qualitatively similar, marked quantitative differences suggest that the subcellular cycling of these two integral membrane proteins occurs by distinct processes. PMID- 2968985 TI - Characterization of junctional and longitudinal sarcoplasmic reticulum from heart muscle. AB - Longitudinal tubules and junctional sarcoplasmic reticulum (SR) were prepared from heart muscle microsomes by Ca2+-phosphate loading followed by sucrose density gradient centrifugation. The longitudinal SR had a high Ca2+ loading rate (0.93 +/- 0.08 mumol.mg-1.min) which was unchanged by addition of ruthenium red. Junctional SR had a low Ca2+ loading rate (0.16 +/- 0.02 mumol.mg-1.min) which was enhanced about 5-fold by ruthenium red. Junctional SR had feet structures observed by electron microscopy and a high molecular weight protein with Mr of 340,000, whereas longitudinal SR was essentially devoid of both. Thus, these subfractions have similar characteristics to longitudinal and junctional terminal cisternae of SR from fast twitch skeletal muscle. Ryanodine binding was localized to junctional cardiac SR as determined by [3H]ryanodine binding. Scatchard analysis of the binding data showed two types of binding (high affinity, Kd approximately 7.9 nM; low affinity, Kd approximately 1 microM), contrasting with skeletal junctional terminal cisternae where only one site with Kd of approximately 50 nM was observed. The ruthenium red enhancement of Ca2+ loading rate in junctional cardiac SR was blocked by pretreatment with low concentrations of ryanodine as reported for junctional terminal cisternae of skeletal muscle SR. The Ca2+ loading rate of junctional cardiac SR was enhanced by preincubation with high concentrations of ryanodine. The apparent inhibition constant (Ki approximately 7 nM) and stimulation constant (Km approximately 1.1 microM) for ryanodine on junctional SR corresponded to the Kd for high affinity binding (Kd approximately 7.9 nM) and low affinity binding (Kd approximately 1.1 microM), respectively. These results suggest that high affinity ryanodine binding locks the Ca2+ release channels in the open state and that low affinity binding closes the Ca2+ release channels of the junctional cardiac SR. The characteristics of the Ca2+ release channels of junctional cardiac SR appear to be similar to that of skeletal muscle SR, but the Ca2+ release channels of cardiac SR are more sensitive to ryanodine. PMID- 2968986 TI - Cell surface heparan sulfate proteoglycan and the neoplastic phenotype. AB - Cell surface proteoglycans are strategically positioned to regulate interactions between cells and their surrounding environment. Such interactions play key roles in several biological processes, such as cell recognition, adhesion, migration, and growth. These biological functions are in turn necessary for the maintenance of differentiated phenotype and for normal and neoplastic development. There is ample evidence that a special type of proteoglycan bearing heparan sulfate side chains is localized at the cell surface in a variety of epithelial and mesenchymal cells. This molecule exhibits selective patterns of reactivity with various constituents of the extracellular matrix and plasma membrane, and can act as growth modulator or as a receptor. Certainly, during cell division, membrane constituents undergo profound rearrangement, and proteoglycans may be intimately involved in such processes. The present work will focus on recent advances in our understanding of these complex macromolecules and will attempt to elucidate the biosynthesis, the structural diversity, the modes of cell surface association, and the turnover of heparan sulfate proteoglycans in various cell systems. It will then review the multiple proposed roles of this molecule, with particular emphasis on the binding properties and the interactions with various intracellular and extracellular elements. Finally, it will focus on the alterations associated with the neoplastic phenotype and will discuss the possible consequences that heparan sulfate may have on the growth of normal and transformed cells. PMID- 2968988 TI - Is renal biopsy necessary in the nephropathic diabetic? PMID- 2968987 TI - The quantification of cerebral infarction following focal ischemia in the rat: influence of strain, arterial pressure, blood glucose concentration, and age. AB - Focal cerebral ischemia was induced by occlusion of the middle cerebral artery in rats. The volumetric assessment of infarcted tissue, 2 days following occlusion, was calculated from the examination of eight preselected coronal sections. Five differing rat strains were examined. A small and variable infarcted volume was seen in Wistar-Kyoto rats; Sprague-Dawley rats had a relatively large, but still variable, infarcted volume. Of the normotensive rat strains, the most reproducible volume of infarcted tissue was seen in Fischer-344 rats; also the absolute value of the infarcted volume did not vary from one series to another in this strain. Chronic arterial hypertension, studied in both normal and stroke prone spontaneously hypertensive rats, was associated with significantly larger infarction volumes. Age does not change the volume of necrosis: Fischer-344 rats were studied at 3, 9, and 20 months of age, and no significant differences were noted between these ages. Experimental diabetes was induced by the administration of streptozotocin 3 days prior to middle cerebral artery occlusion. Severe hyperglycemia (greater than 400 mg/dl) was associated with a considerably increased volume of infarction. The variability of the resultant lesion is high in the most commonly studied strains, but our results suggest that, for studies in normotensive rats, the use of the Fischer-344 strain produces a standardized and repeatable infarction that may be significantly modified by experimental interventions. Age is not a factor that affects the occlusion-induced infarction; in contrast, both chronic arterial hypertension and experimental diabetes aggravate the histological consequences of middle cerebral artery occlusion in the rat. We conclude that quantitative histological evaluation of infarct size allows a meaningful assessment of the gravity of focal cerebral ischemia. PMID- 2968989 TI - Continuous ambulatory peritoneal dialysis (CAPD) in diabetic patients with end stage renal failure in Hong Kong. AB - The authors' experience in managing 17 diabetic patients among their first 100 consecutive patients treated with continuous ambulatory peritoneal dialysis (CAPD) was reviewed. The diabetics were significantly older than the non diabetics, but their biochemistry was comparable to that of the non-diabetics. With three exchanges a day, the requirement for antihypertensives was high (60%). Exit-site infection occurred at a rate of one episode per 9.7 patient-months, and the frequency of peritonitis averaged one episode per 9.4 patient-months; the main culprit was Staphylococcus pyogenes. Rehabilitation was good because the patients had to finance their treatment. In spite of old age, 23% worked full time. There was no progressive increase in serum cholesterol or triglycerides. Glycemic control was good and was comparable whether the patients were given insulin subcutaneously or intraperitoneally. There was a highly significant (p less than 0.001) positive correlation between fasting blood glucose levels and HbA1 concentrations. Fasting blood glucose concentrations did not correlate with either serum cholesterol or triglyceride concentrations. Diabetic retinopathy progressed in five patients, to the point that their vision was severely impaired. There was no relationship between the degree of glycemic control and progression of diabetic retinopathy. Two patients died of cardiovascular causes, but there were no peritonitis-related deaths. Cummulative patient survival at 2 years was 86%, and the corresponding technique survival, 100%. PMID- 2968990 TI - Decreasing insulin requirements in CAPD patients given intraperitoneal insulin. AB - In a retrospective study, insulin requirements were examined in two groups of dialyzed diabetic patients: one treated with subcutaneous insulin and the other with intraperitoneal insulin. In the first group, insulin requirements did not change with time on dialysis irrespective of whether the patients were treated with hemodialysis or continuous ambulatory peritoneal dialysis (CAPD); however, in CAPD patients using the intraperitoneal route of administration, a significant decrease in insulin requirements occurred simultaneously with improved metabolic control. This observation may indicate that intraperitoneal administration of insulin is advantageous as compared to the subcutaneous route. PMID- 2968991 TI - The genesis of the Armanni-Ebstein lesion in diabetic nephropathy. PMID- 2968992 TI - A 2 year evaluation of diabetic patients on continuous ambulatory peritoneal dialysis. AB - Nineteen diabetic patients with end-stage renal disease on CAPD were evaluated over a 2 year period. All but one patient was insulin-dependent, with a mean age of 47.7 years. Average time on CAPD was 16.1 months (range, 2-28 months). Thirteen patients were followed for more than 12 months, and nine for more than 18 months. The mean training period was 22.9 days. Good blood glucose control was obtained with intraperitoneal (IP) insulin in all of the patients. Mean blood glucose levels of 125 +/- 23.08 mg/dl were achieved with 103 +/- 38.5 U/day of regular IP insulin. Glycosalated hemoglobin decreased from a mean of 12.7 +/- 2.35% before CAPD to 10.08 +/- 0.97% during CAPD. Peritoneal creatinine clearance remained stable during the study period, with a concommitant decrease (P less than 0.001) in the mean residual renal creatinine clearance. The incidence of peritonitis was one episode per 7.8 patient-months. Average length of hospitalization was 33.24 days/year. Visual acuity remained stable after 1 year in 73% of the 26 eyes evaluated. No amputations were required in more than 2 years of follow-up. Actuarial survival was 100% at 1 year and 86% at 2 years, and the technique survival of CAPD was 91 and 79%, respectively. These results demonstrate that CAPD is a good dialysis procedure for treating diabetic patients with chronic renal failure, and it offers the advantage of controlling glycemia better than other dialysis methods. PMID- 2968993 TI - The obese Zucker rat model of glomerular injury in type II diabetes. PMID- 2968994 TI - Normoglycemic diabetic retinopathy and nephropathy. AB - End-stage renal failure is one of the major complications of diabetes and a significant cause of death in this population. At present, its cause is unknown, and consequently, attempts to prevent it are arbitrary. It has been suggested that improved control of blood glucose and hypertension may prevent the onset of renal failure in patients with diabetes mellitus. We present a case in which, despite near-normal levels of blood glucose and blood pressure, a relentless downhill course ensued resulting in severe renal failure and near blindness as a result of diabetic nephropathy and retinopathy. PMID- 2968995 TI - Adult polycystic kidney disease with diabetic intercapillary glomerulosclerosis. PMID- 2968996 TI - Diabetic nephropathy: long-term survival. PMID- 2968997 TI - Why do so few diabetic patients attend dialysis centers in central Italy? AB - We report the prevalence of end-stage diabetic nephropathy treated by hemodialysis in the Lazio region, which includes the City of Rome, with a total population of over 5 million people. The percentage of diabetic patients among those attending the dialysis centers was 5% (n = 59), which is below the figures reported for both Northern Europe or Southern Italy (Sicily). A higher number (40%) of patients were affected by non-insulin treated diabetes. This figure is similar to that reported in Southern Italy, but highly discordant from the Northern European data where the large majority of diabetics on hemodialysis are affected by insulin-treated diabetes. Genetic differences and the type of diet may account for the difference in prevalence of end-stage nephropathy between Northern and Southern Caucasians affected by diabetes. PMID- 2968999 TI - The regulation of myocardial perfusion in health and disease. PMID- 2968998 TI - Renal hypertrophy in experimental diabetes: some functional aspects. AB - Glomerular filtration rate (GFR) was measured by constant infusion of iothalamate in conscious rats with and without streptozotocin diabetes. Kidney weight had already increased significantly after 4 days of diabetes. GFR was unchanged at that time, but increased subsequently, so that GFR/kidney weight was normal after 1 month of diabetes. Tubular glucose reabsorption rate was increased to the same extent as kidney weight after 4 days of diabetes. It is concluded that renal growth in experimental diabetes cannot be initiated by glomerular hyperfiltration, but that increased glucose reabsorption may have a causal role. PMID- 2969000 TI - Altered metabolic and hormonal responses to epinephrine and beta-endorphin in human obesity. AB - Catecholamines and endogenous opioid peptides are released in response to stress. Exogenous infusions of epinephrine and beta-endorphin (both in doses of 15, 50, and 80 ng/kg.min sequentially, each dose lasting 30 min) were used to mimic short term stress in both normal weight (body mass index, less than 25 kg/m2) and obese (body mass index, greater than 30 kg/m2) subjects. Fasting plasma insulin, C peptide, and beta-endorphin concentrations were significantly higher in the obese than in the normal subjects (P less than 0.01-0.005). In lean subjects epinephrine produced significant increases in plasma glucose levels, but no appreciable changes in plasma insulin, C-peptide, or glucagon. Infusion of beta endorphin in the same subjects caused plasma glucose and glucagon to rise, but insulin and C-peptide levels did not change. The simultaneous infusion of epinephrine and beta-endorphin produced a glycemic response which, although greater, was not significantly different than the sum of the responses to the individual hormone infusions. However, the two hormones had a synergistic interaction on plasma glucagon levels [total glucagon response, 2275 +/- 370 pg/min.mL (ng/min.L); sum of single effects, 750 +/- 152 (+/- SE) pg/min.mL (ng/min.L); P less than 0.01]. The plasma epinephrine [207 +/- 21, 607 +/- 70, and 1205 +/- 134 pg/mL (1130 +/- 115, 3640 +/- 382, and 6577 +/- 691 pmol/L] and beta-endorphin [875 +/- 88, 1250 +/- 137, and 1562 +/- 165 pg/mL (250 +/- 25, 358 +/- 39, and 447 +/- 47 pmol/L] concentrations attained during the infusions of each single hormone were not different from those recorded during the combined hormonal infusion. In obese subjects epinephrine raised plasma glucose levels and caused dose-related increments of plasma glucagon concentrations. Plasma insulin and C-peptide concentrations remained low and rebounded at the end of the infusions. In the same subjects, beta-endorphin produced elevations of plasma glucose, insulin, C-peptide, and glucagon. When the combined hormonal infusion was given to obese subjects, the plasma epinephrine and beta-endorphin concentrations rose to values not significantly different from those in normal weight subjects. However, there was a dramatic increase in plasma glucose exceeding 200 mg/dL (11.1 mmol/L), which remained elevated 30 min after the infusion. The glucagon response was not greater than the sum of the single effects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969001 TI - Plasma levels of atrial natriuretic hormone in Cushing's syndrome. AB - To examine a possible role for atrial natriuretic hormone (ANH) in the water and electrolyte disturbances associated with hypercortisolism, plasma ANH levels were measured in 18 patients with endogenous Cushing's syndrome. Nine patients had elevated plasma ANH levels compared to normal subjects. The mean plasma ANH concentration [72.5 +/- 13.0 (+/- SE) pg/mL (23.5 +/- 4.2 pmol/L)] in the Cushing's syndrome patients was significantly higher than that in 40 normal subjects [37.6 +/- 1.9 pg/mL (12.2 +/- 0.62 pmol/L)]. A significant positive correlation was found between plasma ANH and cortisol levels in individual patients. There were no significant correlations, on the other hand, between plasma ANH concentrations and PRA, plasma aldosterone levels, or mean blood pressure. After treatment, plasma ANH concentrations decreased in all 6 patients who had elevated plasma ANH levels preoperatively. In 1 patient with Cushing's disease, plasma ANH levels changed in parallel with plasma cortisol concentrations during o,p'DDD treatment. Fifteen patients who were receiving long term synthetic glucocorticoid therapy for the treatment of miscellaneous diseases had a significantly higher mean plasma ANH level [50.2 +/- 4.0 (+/- SE) pg/mL (16.3 +/- 1.3 pmol/L)] than that in normal subjects. These results suggest that plasma ANH levels are elevated in a substantial number of patients with Cushing's syndrome due to either a direct stimulatory effect of glucocorticoid on atrial ANH secretion or, alternatively, intravascular volume expansion resulting from excessive cortisol secretion. PMID- 2969002 TI - Effect of daily and alternate day low dose prednisone on serum cortisol and adrenal androgens in hirsute women. AB - To test the hypothesis that alternate day prednisone treatment more effectively suppresses adrenal androgen secretion (compared to cortisol secretion) than daily prednisone treatment, we measured serum dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), and cortisol concentrations during these two prednisone treatment schedules in eight hirsute women. The women were assigned randomly to receive either a daily nighttime dose of prednisone (100 micrograms/kg) or an alternate nighttime prednisone dose (200 micrograms/kg) for 4 months. During the following 4 months the women received the other schedule. Serum hormone levels were measured 0, 4, and 8 months before and after iv administration of 25 U synthetic ACTH. To optimally compare the daily and alternate day prednisone regimens, hormonal determinations were made on 2 successive days (days 1 and 2) after the last dose of prednisone. We found no evidence for greater suppression of adrenal androgens or lesser suppression of cortisol with alternate day prednisone treatment. Basal serum DHEA and DHEAS concentrations were suppressed to a greater degree than was cortisol during both daily and alternate day prednisone treatments. ACTH-stimulated DHEA and cortisol concentrations were equally suppressed. Only two of the eight women noted improvement in hirsutism during the study, and four women gained weight. Thus, adrenal androgen secretion was more easily suppressed than was cortisol secretion by this low dose of glucocorticoid, but there was no advantage to alternate day therapy. PMID- 2969003 TI - Transient disappearance of immunologic disorders and remission after intercurrent measles infections in children with chronic idiopathic thrombocytopenic purpura. AB - In two children with chronic idiopathic thrombocytopenic purpura (ITP) a transient remission of thrombocytopenia was observed after intercurrent measles infection. Both cases were girls who had a long history of thrombocytopenia. During acute measles infection, the delayed hypersensitivity response was suppressed. Total T lymphocytes, T-cell subsets, especially OKT4 cells, the lymphoproliferative response, and interleukin-2 (IL-2) and gamma-interferon production were decreased accompanying normalization of the OKT4/OKT8 ratio. However, OKT4 cells remained at a reasonably low level and the lymphoproliferative response stimulated with pokeweed mitogen was still in the lower normal range. Direct immunofluorescent study demonstrated that the measles antigen was present in the mononuclear cells, especially OKT4 cells. The levels of platelet-associated IgG antibody (PAIgG) and IgG circulating immune complex (CIC) were undetectable. One month later, the OKT4/OKT8 ratio lymphoproliferative response significantly increased, IL-2 and gamma-interferon production increased, and PAIgG and IgG CIC reappeared with the relapse of thrombocytopenia. There was also a significant increase in in vitro IgG production due to the presence of patient OKT8 cells and/or OKT4 cells. However, there was no enhancement in the presence of patient B cells. This suggests that the presence of specific OKT4 helper T cells and a defect in the suppressor function of suppressor OKT8 cells contribute to an overproduction of IgG and the appearance of PAIgG accompanied by thrombocytopenia. The transient remission associated with measles infection is probably related to the effect of the virus on the helper T cells, resulting in a decrease in specific OKT4 helper T cells and normalization of the OKT4/OKT8 ratio, suppression of IL-2 and gamma-interferon production, and platelet associated IgG production. PMID- 2969004 TI - [Assessment of prognostic factors in patients with liver cirrhosis with emphasis on the usefulness of peritoneoscopy]. PMID- 2969005 TI - LHRH agonists in IVF: different methods of utilization and comparison with previous ovulation stimulation treatments. AB - LHRH agonists are being increasingly used in ovulation stimulation protocols in IVF programmes. We have compared the results of two methods of utilization of LHRH agonists. In the long protocol, gonadotrophin stimulation was only commenced after a preliminary period of pituitary desensitization with LHRH agonist. In the short protocol, exogenous gonadotrophins were administered shortly after the start of LHRH agonist therapy, benefiting from the gonadotrophin flare-up effect. One-hundred-and-eighty-six patients were divided equally between the two treatments. There was no difference in the ovarian response on the day of HCG or the number of mature oocytes recovered. The cleavage rate of mature oocytes was higher in the short protocol (70, versus 56, P less than 0.01). The ongoing pregnancy rate per treatment cycle was similar in both groups (18, in the long protocol and 16, in the short protocol). Analysis of the luteal phases revealed a trend for higher progesterone values in the long protocol although this was only significant on the second day following oocyte retrieval. As the clinical results were similar other factors should be taken into account when deciding therapy. These include patient convenience, cost and side-effects. Other schedules of ovulation stimulation using LHRH agonists are discussed. PMID- 2969006 TI - Syphilis/Lyme/AIDS. PMID- 2969007 TI - Diastolic flow characteristics of severely impaired left ventricles: a pulsed Doppler ultrasound study. AB - Left ventricular inflow was studied with pulsed Doppler echocardiography in 12 normals and 12 patients with a dilated left ventricle as a result of ischemic heart disease. To study the influence of left ventricular relaxation on both the transmitral and apical diastolic flow patterns, we applied range ambiguity. This provides simultaneous display of Doppler shifts obtained at different depths. The diastolic Doppler shift curves at the mitral valve and apical level in normals had the same time of onset, whereas there was an apparent time delay in the patients with heart disease (range 160-360 ms). This might be caused by absence of a normal apical wall motion in these patients. PMID- 2969008 TI - Tendinous intersection as an unusual site for portasystemic anastomosis: an ultrasonographic observation. PMID- 2969009 TI - Morphological aspects of the gingiva in children with Down's syndrome during experimental gingivitis. AB - In a previous investigation, children with Down's syndrome (DS) showed an earlier, more rapid and more extensive gingival inflammation than normal healthy control children. These differences in gingival inflammation may be the result of aberrant morphology of the gingiva related to the genetic disorder in DS children. The aims of the present study were (i) to describe the structural composition of "normal" gingiva in DS compared to control children, (ii) to analyse the histological changes in the gingiva during plaque development and (iii) to investigate whether the clinical findings could be supported by morphological observations. The study was carried out in 8 DS and 8 matched control children. Their ages ranged from 5-10 years. Gingival normality was guaranteed by strict oral hygiene procedures. During a period of 21 days in which oral hygiene was abolished, gingival biopsies were taken from buccal sites of deciduous teeth following a predetermined schedule on days 0, 7, 14 and 21. Results on day 0 showed no morphological differences between the DS and control children regarding oral epithelium, junctional epithelium or connective tissue. During the experimental phase of the study, the amount of plaque accumulation in the DS children gave rise to a more extensive gingival inflammation than in the control children. The gingival inflammation in the DS group started earlier and included: (1) an acute inflammatory response, (2) an increase of the junctional epithelium area, (3) an increase of the infiltrated connective tissue area (ICT) and (4) a decrease in collagen fibre density of about 35-40% compared to day 0. The same phenomena were not seen until 7 days later in the control group. Conversely, the development of a perivascular lymphocyte infiltrate (LI) in the DS children was delayed compared to the control group. This may be caused by the impaired delayed-type hypersensitivity response in DS children. The development of 2 separate infiltrates (ICT and LI) in this age group and the different temporal development of ICT (day 7 for the DS and day 14 for the control group) and LI (day 14 for the DS and day 7 for the control group) does suggest different immunological mechanisms for both areas and both groups. PMID- 2969010 TI - Cellular aspects of and effects on the gingiva in children with Down's syndrome during experimental gingivitis. AB - In a previous experimental gingivitis study, it was shown that in children with Down's syndrome (DS), gingival inflammation started earlier, was more extensive and developed faster, than in normal healthy control children. In both groups, the start of the process was accompanied by an acute inflammatory response and an increase of the infiltrated connective tissue area (ICT). The purpose of the present study was to investigate how these facts were reflected at a cytological level. The study was carried out in 8 DS and 8 matched control children. Their ages ranged from 5-10 years. A "normal" healthy gingiva was attained after strict oral hygiene procedures. During a period of 21 days in which oral hygiene was abolished, gingival biopsies were taken on days 0, 7, 14, and 21. In both groups, junctional epithelium (JE) and ICT contained low numbers of polymorphonuclear leucocytes (PMNs). The start of the inflammation (day 7 for the DS and day 14 for the control children) was marked by a significant positive correlation between the numbers of PMNs in the JE and the ICT, and a significant increase of the numbers of PMNs in ICT. In ICT, a concomitant decrease in collagen fibre density was observed. In the control group, the decrease correlated with the numbers of PMNs in ICT, which suggests that this collagen breakdown is caused by PMN products. After the initial decrease, the collagen fibre density remained fairly constant in this group throughout the study. In the DS group, there was a tendency to a further decrease in the ICT3 area, correlated with the numbers of PMNs in ICT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969012 TI - Multiple Bowen's disease associated with squamous cell carcinoma--a report of a case. PMID- 2969013 TI - Follicular mucinosis: response to indomethacin. PMID- 2969011 TI - Fixed drug eruptions: a study of epidemiological, clinical and diagnostic aspects of 89 cases from India. PMID- 2969014 TI - Griseofulvin-induced photodermatitis--report of six cases. PMID- 2969017 TI - Multiple coronary angioplasty: a model to discriminate systemic and procedural factors related to restenosis. AB - To assess the interrelation of clinical and procedural factors responsible for restenosis, 119 patients undergoing coronary arteriography were studied a mean of 5.8 +/- 3 months after successful multiple percutaneous transluminal coronary angioplasty. In all clinical, angiographic and procedural variables, the 119 patients undergoing repeat catheterization were similar to the 87 patients that did not. Overall, restenosis occurred in 74 (34%) of 215 lesions. Sixty-three patients had no restenosis, 44 had at least one restenosis and 12 had restenosis at all angioplasty sites. The statistical distribution of restenoses did not follow a binomial model, suggesting that restenosis is more than a lesion specific phenomenon. Of all the clinical and procedural variables assessed by multivariate logistic regression analysis, only percent stenosis before angioplasty (p less than 0.01), diabetes mellitus (p less than 0.01) and percent stenosis after angioplasty (p less than 0.05) were predictive of restenosis in the entire group. Patients with no restenosis and patients with restenosis at all sites were not different with respect to procedural variables; however, patients with restenosis at all sites more often (p less than 0.05) had diabetes and recent onset angina. In contrast, patients with no restenosis differed from patients with isolated restenosis with respect to procedural variables: severity of stenosis before and after angioplasty, balloon/artery lumen ratio and maximal inflation pressure. Thus, procedural factors may be more related to isolated restenosis, but patient-related factors such as diabetes and recent onset angina may play a more important role in patients with multiple restenoses. PMID- 2969016 TI - An agoraphobic patient with dental anxiety: report of case. AB - A 46-year-old woman made an appointment for dental therapy. The patient's agoraphobia and dental anxiety presented a difficult treatment situation. An open dialogue between patient and dentist along with a flexible treatment plan and unhurried appointments contributed to the success of the dental rehabilitation. PMID- 2969015 TI - Changing dentists' knowledge, attitudes, and behaviors relating to AIDS: a controlled educational intervention. AB - The efficacy of an educational intervention designed to improve dentists' knowledge, attitudes, and behaviors about acquired immune deficiency syndrome (AIDS) was tested. The intervention had three components: computerized feedback comparing participants' own knowledge, attitudes, and behaviors with those of fellow participants and with an ideal; periodic bulletins; and telephone conference calls with experts. The group receiving the educational package had better scores than a control group on outcomes of willingness to treat persons with AIDS; identification of human immunodeficiency virus lesions; knowledge of AIDS; and completeness of both intraoral and extraoral examinations. It was concluded that intervention is one approach to increasing dentists' positive response to the AIDS epidemic. PMID- 2969018 TI - Change in diameter of coronary artery segments adjacent to stenosis after percutaneous transluminal coronary angioplasty: failure of percent diameter stenosis measurement to reflect morphologic changes induced by balloon dilation. AB - To determine the changes in stenotic and nonstenotic segments of a dilated coronary artery, detailed quantitative angiographic measurements were performed in 342 patients (398 lesions) immediately after angioplasty and at a predetermined follow-up time of 30, 60, 90 or 120 days after the dilation. Measurements of the stenotic segments were expressed as minimal luminal diameter, and the adjacent nonstenotic segments were expressed as interpolated reference diameter (both in millimeters). A follow-up rate of 86% was achieved. In the patients followed up at 30 and 60 days, there was no significant change in either the mean minimal luminal diameter or the mean reference diameter. However, at 90 and 120 days, there was significant deterioration in both the mean minimal luminal diameter (-0.37 and -0.42 mm, respectively) and the mean reference diameter (-0.17 and -0.26 mm, respectively), all of the changes being highly significant (p less than 0.00001). The reference diameter is involved in the dilation process and may be subject to the same restenosis process that takes place in initially stenotic segments. Percent diameter stenosis measurements, which are conventionally used to express the change in the severity of a stenosis after angioplasty, will tend to underestimate the change when there is a simultaneous reduction in the reference diameter. PMID- 2969019 TI - Coronary angioplasty for unstable angina: immediate and late results in 200 consecutive patients with identification of risk factors for unfavorable early and late outcome. AB - Two hundred patients (mean age 56 years, range 36 to 74) with unstable angina (chest pain at rest, associated with ST-T changes) underwent coronary angioplasty. In 65 patients with multivessel disease, only the "culprit" lesion was dilated. The initial success rate was 89.5% (179 of 200 patients). At least one major procedure-related complication occurred in 21 patients (10.5%): (death in 1, myocardial infarction in 16 and urgent surgery in 18). All patients were followed up for 2 years. Five patients died late; 8 had a late nonfatal myocardial infarction and 52 had recurrence of angina pectoris. The restenosis rate was 32% (51 of 158) in the patients with initial successful angioplasty who had repeat angiography. At the 2 year follow-up, after attempted coronary angioplasty in all 200 patients, the total incidence rate of death was 3% (one procedure related; five late deaths), of nonfatal myocardial infarction 12% (16 procedure related and 8 late after angioplasty), and 13% (26 patients) were still symptomatic although they had improved in functional class. Multivariate analysis showed that variables indicating an increased risk 1) for major procedure-related complications were: ST segment elevation, persistent negative T wave and stenosis greater than or equal to 65% (odds ratio 3.7, 3.7 and 3.3, respectively); 2) for angiographic restenosis were: presence of collateral vessels, ST segment depression, multivessel disease, left anterior descending coronary artery stenosis and history of recent onset of symptoms (odds ratio: 2.2, 2.0, 1.9, 1.9 and 0.54, respectively); and 3) for late coronary events (recurrence of angina, late myocardial infarction or late death) were: multivessel disease, total occluded vessel and ST segment elevation (odds ratio 3.7, 2.8 and 0.44, respectively). Thus, coronary angioplasty for unstable angina can be performed with a high initial success rate, but at an increased risk of major complications. The prognosis is favorable after initial successful coronary angioplasty. PMID- 2969020 TI - Left ventricular systolic and diastolic function during acute coronary artery balloon occlusion in humans. AB - Left ventricular function during percutaneous transluminal coronary angioplasty was studied in 16 patients undergoing the procedure. All measurements were performed before and during the first episode of balloon coronary occlusion. In 16 patients (Group A), data were recorded before and 30 or 50 s after balloon inflation, and in 8 of these patients (Group B) data were also recorded 15 min after the complete procedure. Left ventriculograms indicated a marked dyskinesia of the anterior and apical wall in all patients. After balloon inflation, there was a marked depression in stroke index and ejection fraction and an increase in left ventricular end-diastolic pressure and the time constants of relaxation in all patients. Simultaneous recording of left ventricular pressure (Millar micromanometer) during cineangiography permitted the assessment of myocardial and chamber stiffness. Although there was a strong tendency for both myocardial and chamber stiffness to increase after 30 to 50 s of occlusion, these increases were statistically insignificant. In Group B, a third set of angiographic and pressure measurements obtained 15 min after completion of the coronary angioplasty procedure indicated no residual left ventricular dysfunction, and in this respect, the results are of added clinical importance. PMID- 2969021 TI - Guidelines for percutaneous transluminal coronary angioplasty. A report of the American College of Cardiology/American Heart Association Task Force on Assessment of Diagnostic and Therapeutic Cardiovascular Procedures (Subcommittee on Percutaneous Transluminal Coronary Angioplasty). PMID- 2969024 TI - [Lipocortin--a Ca2+-binding protein which has anti-phospholipase A2 activity]. PMID- 2969023 TI - Pneumocystis carinii pneumonia: therapy and prophylaxis. PMID- 2969022 TI - Interaction between fenoterol, ipratropium, and acetylcholine on human isolated bronchus. AB - Functional antagonism between fenoterol (F) and acetylcholine (ACh) and interaction between F and ipratropium (Ipr) on ACh-induced contraction were evaluated on isolated human bronchi. In the presence of increasing concentrations of ACh (2 X 10(-4) and 2 X 10(-3) mol/L), dose-response curves of the relaxant effect of F were shifted to the right (0.45 and 0.92 log units), and the maximal effect of F, expressed as a percentage of the effect of theophylline, 3 X 10(-3) mol/L, was reduced from control values of 92.1 +/- 4% to 70.8 +/- 7.0%, and to 67.2 +/- 7.1%, according to the functional antagonism. In the presence of Ipr, 10(-9) and 10(-8) mol/L, the functional antagonism between ACh and F was partially reversed. Concentration response-curves to F versus ACh, 2 X 10(-3) mol/L, were shifted to the left, and although the -log molar concentration producing 50% of maximal effect was not significantly modified, the maximal effect of F was significantly increased in the presence of Ipr, 10(-8) mol/L. It is concluded that the effect of combined F and Ipr on the isolated human bronchus contracted with ACh appears to be of the additive type. PMID- 2969025 TI - [A successful treatment of left ovarian pregnancy with methotrexate]. PMID- 2969026 TI - Quantifying the effects of spinal manipulations on gait using patients with low back pain. AB - The purpose for conducting this study was to investigate the effects of chiropractic treatment on the gait of a group of subjects with sacroiliac joint syndromes. The clinical results suggest that chiropractic treatments reduce pain, increase mobility of the sacroiliac joint, and restore general functional ability of the patient. Force results obtained by using a force platform during gait of the subjects showed that external forces were significantly different for gait trials executed after chiropractic treatment compared to gait trials executed before chiropractic treatment. Force results were also significantly different for gait trials executed early in the rehabilitation process compared to those executed late in the rehabilitation process. It should now be investigated if and how these changes in external forces influence the internal forces acting on low back structures. Such an investigation may provide further insight into the mechanisms underlying low back problems. PMID- 2969027 TI - Correlates of low back pain in a general population sample: a multidisciplinary perspective. AB - This study identifies correlates of low back pain in a general population sample and defines a profile of subjects with low back pain. A multidisciplinary approach was employed that required surveying and physically assessing 674 subjects on 105 variables in biographical, anatomical, strength and flexibility measurement categories. No attempt was made to select subjects from specific occupational, age, athletic, psychological and anatomical groups or subjects with specific biographical features, which may have resulted in a sample that was atypical of the general population. The results of this study based on a causal comparative ex post facto research design corroborated selected findings of previous research conducted on nongeneral population samples. These findings include relationships between low back pain and age, body type, sex, stress, smoking, selected types of physical activity, occupation and previous injuries to the neck, shoulders, back and upper legs, as well as previous episodes of low back pain. Additional correlates of low back pain that were identified and have little or controversial review in the back literature include: delayed low back pain syndrome caused by abrupt changes in running frequency, Q angle, pes cavus, leg length (right and left), trunk length, genu recurvatum and multiplane strength and flexibility limitations in the hip joints. PMID- 2969028 TI - The endocrinology of cardiovascular control. PMID- 2969029 TI - Effect of serotonin antagonists on prolactin and progesterone secretion in rats: evidence that the stimulatory and inhibitory actions of serotonin on prolactin release may be mediated through different receptors. AB - The serotoninergic regulation of prolactin release was studied in female rats in different reproductive states using ketanserin, a specific S2 receptor blocker, ICS 205-930 ((3 alpha-tropanyl)1H-indol-3-carboxylic acid ester), a specific S3 receptor blocker and p-chlorophenylalanine (pCPA), a serotonin synthesis inhibitor. Administration of ketanserin to pro-oestrous rats inhibited the afternoon prolactin surge; this inhibition was prevented by progesterone. On day 3 of pregnancy, pCPA or ketanserin blocked the afternoon prolactin surge, and administration of oestrogen (on day 2) and progesterone (on day 3) in combination, but not alone, prevented this effect. On day 9 of pregnancy, treatment with oestrogen (on day 8) and progesterone (on day 9) induced an afternoon surge of prolactin which was prevented by administration of ketanserin or pCPA. On days 9 and 16, pCPA induced a slight increase in serum prolactin in rats not treated with steroids, but ketanserin had no effect. On day 13, ketanserin and pCPA had no effect on serum prolactin levels, but after increasing serotoninergic transmission by injecting fluoxetine and 5-hydroxytryptophan, serum prolactin levels were decreased. On day 19, ketanserin produced a transient increase in the serum concentration of prolactin, probably produced by the marked decrease in the serum concentration of progesterone induced by the S2 receptor blocker. Administration of ICS 205-930 to pro-oestrous rats or rats on day 19 of pregnancy had no effect on serum concentrations of prolactin and progesterone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969030 TI - Effects of bovine follicular fluid on gonadotrophin secretion in intact and chronically ovariectomized ewes before and after desensitization of pituitary gonadotrophs to gonadotrophin-releasing hormone. AB - Intact and chronically ovariectomized ewes were treated for 4 days with charcoal treated bovine follicular fluid (FF) or charcoal-treated bovine serum during the late-anoestrous period, and the effects on basal and gonadotrophin-releasing hormone (GnRH)-induced secretion of LH and FSH observed. Subsequently, ewes received s.c. implants containing a sustained-release formulation of a potent GnRH agonist D-Ser(But)6-Azgly10-LHRH (ICI 118630) to desensitize pituitary gonadotrophs to hypothalamic stimulation, and the effects of bovine FF and bovine serum were re-assessed 2 weeks later. Chronic exposure (for 2-3 weeks) to ICI 118630 significantly reduced basal levels of LH and FSH in both intact and ovariectomized ewes and completely abolished both spontaneous LH pulses as well as exogenous GnRH-induced acute increases in plasma LH and FSH levels. Treatment with bovine FF significantly reduced plasma FSH levels, but not LH levels, in both intact and ovariectomized ewes before and after chronic exposure to ICI 118630. In intact ewes before exposure to ICI 118630, treatment with bovine FF actually enhanced pulsatile LH secretion and raised mean plasma LH levels by 240% (P less than 0.05). No such stimulatory effect of bovine FF on LH secretion was observed in intact ewes exposed to ICI 118630 or in ovariectomized ewes before or after exposure to ICI 118630, suggesting that the effect probably involved an alteration in ovarian steroid feedback affecting hypothalamic GnRH output. Treatment with bovine FF did not significantly affect the magnitude of GnRH induced surges of LH or of FSH observed in either intact or ovariectomized ewes before exposure to ICI 118630.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969031 TI - Dynamics of activation in semantic and episodic memory. AB - Spreading-activation models for the structure of semantic and episodic memory postulate a network of interconnected nodes in which activation spreads from a source node to recipient nodes. These models account for a broad range of memory related processes, including word recognition, sentence verification, prose comprehension, and sentence production. A fundamental question regarding this account concerns the nature of activation growth at each node in the network. Two mutually exclusive possibilities are (a) that activation grows in a discrete fashion, making abrupt transitions between two or more distinct states and (b) that activation grows continuously from a resting level to an asymptotic level. In the present article, we characterize this dichotomy with examples from the literature, and we apply an adaptive priming procedure for testing discrete versus continuous activation models. Our procedure involves the presentation of prime stimuli at various moments before a test stimulus; subjects are required to make a lexical (word/nonword) decision about the test stimulus. The duration of the interval between the prime and test stimuli is varied adaptively on the basis of subjects' performance. Reaction times are recorded as a function of this duration. According to discrete activation models, there is a unique reaction time distribution associated with each possible state of node activation. The distribution of reaction times observed when the test stimulus appears near the moment of transition between discrete states should therefore constitute a finite mixture of the underlying basis distributions associated with the individual discrete activation states. The mixture proportion will depend on the relation between the priming interval and the distribution of state-transition times. Continuous activation models assert instead that activation grows continuously over time and that there is a unique reaction-time distribution associated with any given degree of intermediate priming. Such models predict that no finite mixture distribution will emerge when the priming interval has a fixed intermediate duration. Two experiments with the adaptive priming procedure are reported to test these alternative predictions. In Experiment 1, the prime and test stimuli were semantically associated words (e.g., bread-butter). In Experiment 2, episodic associations between the prime and test stimuli were established through paired associate learning. For both cases, the mixture prediction failed, and two-state discrete activation models were rejected.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969033 TI - Minimodularity and the perception of layout. AB - In natural vision, information overspecifies the relative distances between objects and their layout in three dimensions. Directed perception applies (Cutting, 1986), rather than direct or indirect perception, because any single source of information (or cue) might be adequate to reveal relative depth (or local depth order), but many are present and useful to observers. Such overspecification presents the theoretical problem of how perceivers use this multiplicity of information to arrive at a unitary appreciation of distance between objects in the environment. This article examines three models of directed perception: selection, in which only one source of information is used; addition, in which all sources are used in simple combination; and multiplication, in which interactions among sources can occur. To monocular spatial information, using all combinations of the presence or absence of relative size, height in the projection plane, occlusion, and motion parallax. Visual stimuli were computer generated and consisted of three untextured parallel planes arranged in depth. Three tasks were used: one of magnitude estimation of exocentric distance within a stimulus, one of dissimilarity judgment in how a pair of stimuli revealed depth, and one of choice judgment within a pair as to which one revealed depth best. Grouped and individual results of the one direct and two indirect scaling tasks suggest that perceivers use these sources of information in an additive fashion. That is, one source (or cue) is generally substitutable for another, and the more sources that are present, the more depth is revealed. This pattern of results suggests independent use of information by four separate, functional subsystems within the visual system, here called minimodules. Evidence for and advantages of minimodularity are discussed. PMID- 2969032 TI - The time course of repetition effects for words and unfamiliar faces. AB - The repetition effect on reaction time to words and unfamiliar faces was examined at lags of 0, 4, and 15 items between first and second presentations. For words, subjects made either a lexical decision or a decision based on the stimulus's structural attributes. In the lexical decision task, a significant repetition effect was found at all three lags for words, whereas for nonwords the effect was significant only at Lag 0. In the structural decision task, the repeated decision was facilitated for both words and nonwords only at Lag 0, despite a word superiority effect at all lags. Target faces were presented either zero, one, or five times before testing. Subjects made either structural discriminations (face/nonface) or recognition judgments. In the structural discrimination task, the effect of repetition was significant only at Lag 0 (regardless of the number of pretest presentations). In the recognition task, the repetition effect was longer lasting, and its magnitude increased with the number of presentations which, presumably, determined the strength of the episodic memory trace. These results are taken as showing that repetition effects, like other measures of memory, are influenced by the type of stimulus, its preexperimental history, the level to which it is processed, and the lag between the initial presentation and the test. The manner in which these variables affect performance, however, may differ across memory tests. The dissociations between performance among repetition tests, and between repetition tests and other types of memory tests, is interpreted according to a task-specific, component-process approach to memory. PMID- 2969034 TI - The acquisition of task-specific productions and modifications of declarative representations in spatial-precuing tasks. AB - The stimulus-response translation stage of human information processing plays a mediating role of relating stimuli to assigned responses. The translation stage has been implicated as the locus of a pattern of differential precuing benefits obtained in spatial-choice tasks (Proctor & Reeve, 1986; Reeve & Proctor, 1985): When pairs of finger responses from the middle and index fingers of each hand are precued, the two leftmost and two rightmost responses show the greatest benefit. This pattern of differential benefits, which occurs regardless of whether the hand placement is adjacent or overlapped, has been attributed to spatially coded representations of the stimulus and response sets in the translation stage. Experiment 1 evaluated whether the mediating role of the translation stage changes with practice. All precued pairs of responses showed equivalent benefits in the last of three sessions. This result indicates that the spatial representations used initially to translate between stimuli and responses have been altered to be more efficient or have been replaced by productions that directly specify fingers. Experiment 2 used a fourth session in which subjects were transferred from the overlapped hand placement to the adjacent placement, or vice versa. For subjects in the former condition, the pattern of differential precuing benefits reappeared in the transfer session. This lack of transfer is consistent with the hypothesis that task-specific productions develop with practice that directly relate stimuli to fingers. For subjects who practiced with the adjacent placement and switched to the overlapped placement, only a nonsignificant tendency existed for the pattern of differential precuing benefits to reappear. This failure of the pattern to reappear could indicate that spatial representations continue to be used to translate between stimuli and responses. Alternatively, as occurs with the overlapped placement, task-specific productions could be acquired that relate stimuli to fingers. If so, the failure of the pattern of differential precuing benefits to reappear would reflect a modification in the representations that are used for translation in the transfer session. Specifically, if subjects were coding the stimulus and response sets on the basis of the distinction between the two hands, as well as the spatial distinction, the differential benefits would be minimized because hand coding should benefit different responses from those benefitted by spatial coding. These alternative explanations were evaluated in Experiment 3 by having subjects who practiced with the adjacent placement switch to a placement in which the hands were crossed completely.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969035 TI - Properties of standard avian slow muscle grafts following long-term regeneration. AB - The tonic anterior latissimus dorsi (ALD) of adult pigeons was orthotopically homografted and evaluated after 11 months of regeneration for histological, histochemical, electromyographic (EMG), and mechanical properties. The resting EMG activity of the grafts was lower in amplitude than that of the controls, but showed the tonic pattern typical for these tonic muscles. The control and grafted muscles had a histochemically homogeneous population of fibers with moderate myofibrillar adenosine triphosphatase activity. Succinic dehydrogenase activity was moderate for the control muscles, but low for the grafts. The regenerated muscles had fewer and smaller fibers and had much larger intersynaptic distances. Both the regenerated and the contralateral control muscles were slow contracting and maintained tetanic tension for prolonged periods with direct electrical stimulation. The relaxation was slower in the grafted muscle than in the control. The grafts produced 40% of the maximum tension of the control muscles, but the rate of tension development was similar between the two groups. The results indicate that the tonic properties were regenerated, but the innervation pattern was altered and the grafted muscles did not have normal mature fibers even after long-term regeneration. PMID- 2969036 TI - Lipids and beta-thromboglobulin in patients with pheochromocytoma. PMID- 2969037 TI - Electrocardiographic findings in hypertensive patients of a population sample. Role of sex, age, and antihypertensive treatment. AB - During the initial phase of the World Health Organization (WHO) "Community Control Program of Hypertension" in Italy, 1190 subjects with high blood pressure derived from a general population random sample (5856 people of both sexes, aged 20 to 64 years) were enrolled in a hypertension register. At the registration visit (RV), each of them gave case history details and underwent a complete clinical examination, blood and urine tests, and a standard 12-lead electrocardiogram (ECG). All the ECGs were read by a single coder, using the second version of the Minnesota Code (MC). We calculated the prevalence of electrocardiographic codes according to sex, age, and the state of antihypertensive treatment. The overall prevalence of electrocardiographic abnormalities (i.e., all codes except 1:0 and isolated 9:4) was 40.8% with a slightly higher prevalence in males than in females: 42.4% versus 39.4%. Codes related to left ventricular hypertrophy (LVH) (3:1 or 3:3) were also more frequent in males (21.2%) than in females (14.5%) but not those related to ischemia (4:1-4:3 or 5:1-5:3). In fact, group 4 codes were present in 4.0% of males and 16.1% of females; group 5 codes, in 5.7% of males and 18.1% of females. Abnormal codes generally increased with increasing age, but those related to LVH did not follow this general rule in males. In fact, in the age class 20-29 years, codes 3:1 were found in 11.1% and codes 3:3 in 17.5% of the subjects, whereas the corresponding frequencies in the oldest age group (60-64 years) were 15.2% and 12.4%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969039 TI - Hypertensive complications and home blood pressure: comparison with blood pressure measured in the doctor's office. AB - We have compared hypertensive target organ damage with home blood pressure readings (HBPs) and with office blood pressure readings (OBPs) in 100 patients with mild to moderate essential hypertension. The correlation between blood pressure levels and hypertensive target organ damage in HBPs and OBPs were similar (r = .42, p less than 0.001 for systolic HBPs; r = .33, p less than 0.001 for diastolic HBPs; r = .42, p less than 0.001 for systolic OBPs; r = .34, p less than 0.001 for diastolic OBPs). In most instances, HBPs were lower than corresponding OBPs. Among individual patients whose OBPs were identical, HBPs in some instances differed strikingly. Optic fundi abnormalities were significantly more severe in patients whose systolic HBPs were 150 mmHg or greater, than in those whose systolic HBPs were less than 150 mmHg (p less than 0.05). Hypertensive complications did not differ among office hypertensive patients who were normotensive or borderline hypertensive at home, from the differences of OBPs. We concluded that overall hypertensive complications were equally related to HBPs and OBPs, but patients with discrepancies between HBPs and OBPs had fewer hypertensive complications. Thus, both OBPs and HBPs should be considered in deciding therapy. PMID- 2969038 TI - Antihypertensive and hormonal effects of PN 200-110, a new calcium-channel blocker, in essential hypertension. AB - Twenty-four patients (aged 34-68), with mild to moderate essential hypertension, received 10 weeks of treatment with the new calcium-channel blocker PN 200-110. Eighty-six percent of the patients achieved goal blood pressure (i.e., sitting diastolic pressure less than or equal to 90 mmHg). The average reduction in sitting and standing systolic and diastolic blood pressure (-14 +/- 2/-15 +/- 1 mmHg, -14 +/- 3/-13 +/- 2 mmHg, respectively) was highly significant (p less than 0.001). No reflex tachycardia (sitting pulse 1 +/- 2 bpm; standing pulse 3 +/- 2 bpm) or weight change (-0.3 +/- 0.9 pounds) accompanied the hypotensive response. Although the fall in blood pressure was accompanied by a small but significant increment in plasma renin activity (0.5 +/- 0.2 ng/ml/hr [p less than 0.05]), no significant changes in plasma angiotensin II, aldosterone, or prostaglandin E2 metabolite occurred. There was no significant correlation between the fall in blood pressure and baseline (pretreatment) or stimulated (posttreatment) levels of plasma renin activity, angiotensin II, aldosterone, or prostaglandin E2 metabolite. PMID- 2969040 TI - Comparison of technetium-99m MAG3 with iodine-131 hippuran by a simultaneous dual channel technique. AB - Technetium-99m MAG3, a technetium-labeled analog of hippuran, was compared with [131I] hippuran using a simultaneous dual isotope study in 20 patients. The plasma clearance for MAG3 was lower than that of hippuran, but its plasma concentration was higher, resulting in similar rates of excretion and similar renal time-activity curves. Apart from better statistics with the technetium labeled agent, there were no clinically significant differences in this group of patients. PMID- 2969041 TI - Thallium-201/technetium-99m-RP-30A disparity in the course of myocardial infarction after attempted reperfusion. AB - Recent reports have established that 201Tl may be taken up in areas of recent myocardial infarction after myocardial blood flow is re-established. In addition, there is accelerated 201Tl "washout" from these regions producing a pattern of "reverse redistribution." We present a case in which these phenomena may have contributed to a disparity of findings between a 201Tl stress imaging study and a repeat stress imaging study performed with a [99mTc] isonitrile (99mTc-RP-30A). PMID- 2969042 TI - Effect of nicotinamide on methionine metabolism in rat liver. AB - To test the response to increased utilization of methyl groups, we administered large dosages of nicotinamide to rats fed an adequate diet that contained limited amounts of methionine and choline. During the 4 d after the injection, we observed several significant effects on the hepatic concentrations of the enzymes and metabolites of methionine metabolism. Methionine and S-adenosylmethionine remained at control levels; the concentrations of S-adenosylhomocysteine exceeded the control values from 4 to 16 h; and the levels of serine and betaine were lower after 16 h. Treatment with nicotinamide resulted in higher hepatic levels of methionine adenosyltransferase (after 4 h) and cystathionine synthase (after 16 h). These data indicate that increases in both homocysteine methylation and S adenosylmethionine synthesis may be components of the response to excessive methyl group consumption. An increased synthesis of cystathionine would provide for the removal of S-adenosylhomocysteine (and homocysteine) derived from the adenosylmethionine-dependent methylation of nicotinamide. PMID- 2969043 TI - Use of workers' compensation claims data for surveillance of cumulative trauma disorders. AB - Workers' compensation claims in Ohio were evaluated as a source of surveillance data for identifying workplaces at high risk of cumulative trauma disorders (CTDs) and analyzed for their demographic and industrial characteristics. During a 5-year period (1980 to 1984), 6,849 workers' compensation claims met the case criteria for CTDs. Tenosynovitis due to continuous motion was the most frequently reported condition (58%), and the wrist was the body part most frequently affected (48%). The highest case rate was observed for female workers in the 36 to 45 age group. Incidence rates for individual companies were determined and those with the highest rates for CTDs were identified. The employer-specific rates for CTDs based on workers' compensation claims data can be used as an effective surveillance tool in locating high-risk operations where ergonomic interventions can be implemented to reduce CTD hazards. PMID- 2969044 TI - Working disability due to occupational back pain: three-year follow-up of 2,300 compensated workers in Quebec. AB - A cohort of 2,342 cases constituting a random sample of all occupational back injuries compensated in Quebec (Canada) during 1981 was observed prospectively for 3 years to study associations between cumulative duration of absence from work and sex, age, site of symptoms (cervical, thoracic, and lumbar), and occupation. Of the cohort members, 227 (9.7%) cumulated 6 months of absence or more. A logistic regression model showed age and site of symptoms to be the two most important risk factors associated with absences of 6 months or more. No association was found with sex or occupation. The results showed discrepancies between measures of frequency distribution of back injuries and duration of absence from work, an important finding in terms of identifying health priorities. PMID- 2969045 TI - Occupational vitiligo due to unsuspected presence of phenolic antioxidant byproducts in commercial bulk rubber. AB - We investigated the occurrence of cutaneous depigmentation (vitiligo) among employees of a company that manufactured hydraulic pumps. The interiors of these pumps were injection-molded with rubber. We identified a small but significant cluster of vitiligo cases among a group of employees who frequently handled the rubber used in this injection molding process. Although none of the additives specified in the rubber formulations was a phenolic or catecholic derivative, known to be potential causes of chemically induced vitiligo, gas chromatographic analysis identified a para-substituted phenol (2,4-di-tert-butylphenol, DTBP) in solid samples of the most frequently used rubber. Surface wipe analysis confirmed that workers could be exposed to DTBP from simple handling of the rubber. We subsequently established that the solid bulk rubber used as the base in these stock rubber formulations contained both DTBP and smaller quantities of p-tert butylphenol. Both had formed as unsuspected byproducts during chemical synthesis of two antioxidants added to the solid bulk rubber by a major rubber supplier. We conclude that the unsuspected presence of potential chemical depigmenting agents in solid bulk rubber, from which industrial rubber products are formulated, may contribute to the occurrence of occupational vitiligo, and that a simple review of ingredients in rubber formulations is inadequate to detect their presence. PMID- 2969047 TI - Subacute infantile mountain sickness. AB - A description is given of a disease of infants occurring in Lhasa, Tibet at an altitude of 3600 m. Typically if affects infants who have been born at low altitude and subsequently brought to residue in Lhasa, and it is usually fatal within a few weeks or months. There is extreme medial hypertrophy of muscular pulmonary arteries and muscularization of pulmonary arterioles, together with dilatation of the pulmonary trunk and massive hypertrophy and dilatation of the right ventricle. The disease is distinct from acute or chronic mountain sickness and appears to be the human counterpart of 'brisket disease' in cattle. PMID- 2969048 TI - AIDS: dental implications. PMID- 2969046 TI - The immunohistology of synovial lining cells in normal and inflamed synovium. AB - The immunohistology of synovial lining cells (SLCs) in normal and inflamed hyperplastic synovium was investigated using monoclonal antibodies directed against leucocyte common antigen (LCA) HLA-DR and other macrophage components. We found that some SLCs in normal synovium express LCA, HLA-DR, and monocyte/macrophage-associated antigens. The number of SLCs expressing these antigens is increased in hyperplastic osteoarthritic (OA) and rheumatoid (RA) synovium. Some SLCs which did not react for LCA or other macrophage markers but were positive for HLA-DR were also noted in normal synovium and some segments of hyperplastic OA synovium. SLCs which are positive for LCA, HLA-DR, and macrophage markers contribute to the intimal hyperplasia in RA where they account for the majority of SLCs in the synovial intima. In OA synovium, the distribution of SLCs showing this pattern of reactivity was less uniform with numerous SLCs which were positive for HLA-DR but negative for LCA and other macrophage markers also present in the synovial intima. These findings indicate that there are some SLCs of bone marrow origin in normal and hyperplastic synovium. They also suggest that recruitment of SLCs of marrow origin is important in the production of intimal hyperplasia in both RA and OA and that there is also a significant local proliferation of non-marrow derived SLCs in OA. PMID- 2969049 TI - Effects of milrinone on Ca++-sensitivity of myofibrillar Mg-adenosine triphosphatase isolated from normal human and canine hearts. AB - Sensitization or desensitization of cardiac actomyosin to calcium has been demonstrated with several pharmacological agents. The effect of milrinone on the sensitivity of cardiac Mg-adenosine triphosphatase (ATPase) activity to calcium was studied in purified myofibrils isolated from normal human hearts (after accidental death or trauma that caused no cardiac damage as established by the attending physician) and from normal canine hearts (established by echocardiography), over a range of calcium concentrations (pCa, 8 to 5). Caffeine, a cardiac stimulant that has been shown to increase the sensitivity of myofibrillar Mg-ATPase activity to calcium in rat ventricle, was used in this study to establish its effect on canine and human myofibrils in comparison with that of milrinone. Caffeine, at concentrations of 40 mM, caused statistically significant sensitization of canine and human myofibrils to calcium. In canine myofibrils, the calcium-dependent Mg-ATPase activity increased from 11.0 +/- 1.2 to 18.8 +/- 2.6 nmol of Pi per mg of protein per min at pCa 6.73 (N = 9, P less than .05) and from 32.9 +/- 2.1 to 37.3 +/- 2.2 nmol of Pi per mg of protein per min at pCa 6.16 (N = 9, P less than .05), whereas total Mg-ATPase activity increased from 23.4 +/- 1.5 to 33.6 +/- 2.6 nmol of Pi per mg of protein per min at pCa 6.73 (N = 9, P less than .05) and from 45.2 +/- 2.2 to 52.2 +/- 2.5 nmol of Pi per mg of protein per min at pCa 6.16 (N = 9, P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969051 TI - Autocannibalization of surviving and necrotic rat groin flaps. AB - Autocannibalization of the rat groin flap was studied one week after a long ischemic period. It was shown that nearly all necrotic flaps were autocannibalized, but surviving flaps were not. The result was similar, whether the epigastric nerve was cut or kept intact. Against this background, it is emphasized that autocannibalized flaps should not be excluded from experimental studies, as this may alter the results. PMID- 2969052 TI - Infanticide for handicapped infants: sometimes it's a metaphysical dispute. AB - Since 1973 the practice of infanticide for some severely handicapped newborns has been receiving more open discussion and defence in the literature on medical ethics. A recent and important argument for the permissibility of infanticide relies crucially on a particular concept of personhood that excludes the theological. This paper attempts to show that the dispute between the proponents of infanticide and their religious opponents cannot be resolved because one side's perspective on the infant is shaped by a metaphysics that is emphatically rejected by the other. In such a situation philosophical argument is powerless to bring about a resolution because there can be no refutation of one side by the other. PMID- 2969050 TI - Fibre types, calcium-sequestering proteins and metabolic enzymes in denervated and chronically stimulated muscles of the rat. AB - 1. Fibre types, Ca2+-sequestering proteins (parvalbumin, sarcoplasmic reticulum Ca2+-ATPase), enzyme activities of energy metabolism, and lactate dehydrogenase isozymes were studied in extensor digitorum longus (EDL) and soleus muscles of adult rats after denervation and direct stimulation for 45-61 days. The stimulation resembled the firing patterns of normal motor units (Hennig & Lomo, 1985) in either soleus (20 Hz, high amount) or EDL (150 Hz, low and high amount). 2. Low- and high-amount 150 Hz stimulation maintained essentially normal properties in the denervated EDL and induced many EDL-like properties in the denervated soleus, e.g. pronounced increases in Ca2+-ATPase and parvalbumin contents, a rise in the glycolytic enzyme activities, as well as a reversal of the lactate dehydrogenase H/M subunit ratio. The 150 Hz high-amount stimulation was not as efficient as the 150 Hz low amount in maintaining the fast properties of denervated EDL with the exception of the enzymes of aerobic oxidative metabolism. High-amount 20 Hz stimulation maintained essentially normal properties in the soleus but induced only moderate slow muscle characteristics in the EDL. 3. The maintenance of essentially normal properties by 'native' and the changes induced by 'foreign' stimulus patterns in the absence of the nerve indicate that evoked muscle activity plays a major role in controlling the phenotypic expression of muscle properties. However, the different responses of the denervated fast- and slow-twitch muscles to identical stimulus patterns further suggest that rat EDL and soleus muscles contain intrinsically different muscle fibres. PMID- 2969053 TI - Tumor-inhibitory antibiotic uptake facilitated by leukoregulin: a new approach to drug delivery. AB - The uptake of tumor-inhibitory antibiotics by human K562 erythroleukemia cells was examined in the presence of leukoregulin to determine if the lymphokine's ability to increase the plasma membrane permeability of tumor cells facilitates concurrent entry of pharmacologically active molecules. K562 cells were exposed to 0.25-2.0 micrograms of doxorubicin/mL for up to 60 minutes at 37 degrees C. Commencing within 15 minutes, leukoregulin increased the entry of doxorubicin approximately twofold and the uptake of mitomycin, mithramycin, and propidium iodide twofold to tenfold. This finding indicates the potential biotherapeutic value of leukoregulin in promoting the selective entry of pharmacologically active molecules into leukoregulin-sensitive target cells. PMID- 2969055 TI - The gene encoding the nonstructural protein of B19 (human) parvovirus may be lethal in transfected cells. AB - The B19 parvovirus is a cause of bone marrow failure in humans. B19 is toxic to erythroid progenitor cells in vitro. Viral products possibly responsible for toxicity were explored by transfection of cloned B19 genome into HeLa cells. The nonstructural (NS) protein was detected in cells 30 h after transfection. Plasmids containing the B19 genome were transfected with selectable marker genes in stable transformation assays. Plasmids that contained the left side of the B19 genome, which encodes the NS protein of the virus, inhibited antibiotic-resistant colony formation. Transformation occurred when NS protein expression was blocked by mutation. Suppression of transformation by NS protein was not tissue specific, suggesting a role for NS protein in toxicity for nonpermissive cells without parvovirus replication or virion accumulation. PMID- 2969054 TI - Generation and characterization of a temperature-sensitive mutation in the NS-1 gene of the autonomous parvovirus minute virus of mice. AB - In-phase single-codon insertion mutations were constructed in the open reading frames of the NS-1 and NS-2 genes of the autonomous parvovirus minute virus of mice. A viral mutant containing an isoleucine insertion exclusively within NS-1 between residues 229 and 230 was isolated that produced approximately 3 orders of magnitude fewer plaques at 39 degrees C than at 32 degrees C. Preliminary characterization of the mutant demonstrated that the NS-1 gene product is independently required for both genome amplification and the regulation of the temporal expression between the two viral transcription units during lytic infection. PMID- 2969056 TI - Purification and functional properties of simian virus 40 large and small T antigens overproduced in insect cells. AB - The insect baculovirus Autographa californica nuclear polyhedrosis virus was used as an expression vector for the simian virus 40 (SV40) small t (t) and large T (T) antigens. Spodoptera frugiperda (SF9) cells infected with recombinant viruses encoding these proteins produced approximately 1 to 2 micrograms of t and up to 30 micrograms of T per 3 X 10(6) cells. The former was highly soluble after Nonidet P-40 extraction of the infected cells, unlike its Escherichia coli produced counterpart. Both SF9-produced proteins were of authentic size and could be readily immunoprecipitated by specific antibodies. Single-step immunoaffinity chromatography was used to purify the two proteins to near homogeneity, with yields averaging 70% in each case. Experiments to test the biological activity of the baculovirus SV40 proteins showed that SF9 t was capable of associating with two of the cellular proteins reported to bind to t in SV40-infected mammalian cells. Moreover, SF9 T had ATPase activity comparable to that of T produced in monkey cells, exhibited helicase activity and SV40 origin-specific DNA binding, and was active in the SV40 DNA replication assay in vitro. Thus, the SV40 T antigens produced in insect cells can be used in future studies of their biochemical roles in vitro and in vivo. PMID- 2969057 TI - Two nuclear location signals in the influenza virus NS1 nonstructural protein. AB - The NS1 protein of influenza A virus has been shown to enter and accumulate in the nuclei of virus-infected cells independently of any other influenza viral protein. Therefore, the NS1 protein contains within its polypeptide sequence the information that codes for its nuclear localization. To define the nuclear signal of the NS1 protein, a series of recombinant simian virus 40 vectors that express deletion mutants or fusion proteins was constructed. Analysis of the proteins expressed resulted in identification of two regions of the NS1 protein which affect its cellular location. Nuclear localization signal 1 (NLS1) contains the stretch of basic amino acids Asp-Arg-Leu-Arg-Arg (codons 34 to 38). This sequence is conserved in all NS1 proteins of influenza A viruses, as well as in that of influenza B viruses. NLS2 is defined within the region between amino acids 203 and 237. This domain is present in the NS1 proteins of most influenza A virus strains. NLS1 and NLS2 contain basic amino acids and are similar to previously defined nuclear signal sequences of other proteins. PMID- 2969058 TI - Immunization of mice with dengue structural proteins and nonstructural protein NS1 expressed by baculovirus recombinant induces resistance to dengue virus encephalitis. AB - We have constructed a recombinant baculovirus containing a 4.0-kilobase dengue virus cDNA sequence that codes for the three virus structural proteins, capsid (C) protein, premembrane (PreM) protein, and envelope glycoprotein (E), and nonstructural proteins NS1 and NS2a. Infection of cultured Spodoptera frugiperda cells with this recombinant virus resulted in the production of E and NS1 proteins that were similar in size to the corresponding viral proteins expressed in dengue virus-infected simian cells. Other dengue virus-encoded proteins such as PreM and C were also synthesized. Rabbits immunized with the dengue virus protein products of the recombinant virus developed antibodies to PreM, E, and NS1, although the titers were low, especially to PreM and E. Nevertheless, the dengue virus antigens produced by the recombinant virus induced resistance in mice to fatal dengue encephalitis. PMID- 2969059 TI - Leads from the MMWR. Changing patterns of groups at high risk for hepatitis B in the United States. PMID- 2969060 TI - Medroxyprogesterone acetate and sleep apnea. AB - Seven patients with sleep apnea DOES and one with sleep apnea DIMS were treated with medroxyprogesterone acetate (MPA). The therapeutic effect was confirmed in most of them by polysomnographic recording. A marked increase of TST was observed in two patients whose AI and/or %SAT remarkably decreased. Contrarily, three patients exhibited a considerable decrease in TST, and AI and/or %SAT were reduced remarkably in two of them with MPA. After the MPA medication a few patients complained of disturbed nocturnal sleep. A significant positive correlation was observed between the decreased rate of TST and that of the mean duration of apneas. From these results, it was considered that MPA has a mild activating action on the arousal system in the CNS, and that the action may be partly responsible for the therapeutic effects of MPA on sleep apneas. PMID- 2969061 TI - Lp(a) lipoprotein in cerebrovascular disease and dementia. AB - Lp(a) lipoprotein has been considered an independent risk factor in the development of coronary heart disease (CHD). We examined the role of Lp(a) in patients with cerebrovascular disease (CVD) and those with dementia. The Lp(a) concentration in patients with CHD, those with cerebral infarction due to a large artery occlusion and those with vascular dementia (VD) was significantly higher than that of age-matched control subjects. However, the Lp(a) concentration was not high in cerebral infarction due to a small artery occlusion, intracerebral hemorrhage and dementia of the Alzheimer type (DAT). The present results suggest that Lp(a) should cause VD as well as CVD, and that Lp(a) should be one of the indicators that distinguish VD from DAT. PMID- 2969063 TI - [Phospholipid composition of high density lipoprotein subclasses in patients with ischemic heart disease in Tashkent]. PMID- 2969062 TI - Cell-mediated cytotoxic activity of spleen cells from patients with gastric carcinoma. AB - The cell-mediated cytotoxic activities of cells from the spleens (SP cells) of patients with gastric carcinoma were assayed in comparison with the activities of peripheral blood mononuclear cells (PBM cells) from the same patients, and from patients with benign lesions. The natural killer cell (NK) activity of the SP cells and their capacity to generate allogeneic cytotoxicity in mixed lymphocyte culture (MLC) were very similar to those of the PBM cells. The cytotoxic activity of SP cells induced by alloactivation in MLC, however, was significantly higher than that of the PBM cells from the same patient as well as from patients with benign lesions. The production of interleukin 2 (IL 2) and the ability to induce cytotoxic cells after activation with IL 2 (LAK) were therefore examined. Both the ability to produce IL 2 and to generate LAK cells were shown to be significantly increased in SP cells when compared to PBM cells. These results indicate that the spleen may be a potential reservoir for the precursors of these activated killer cells in patients with gastric carcinoma. Furthermore, it may play an important role in the defence against tumors in these patients. PMID- 2969064 TI - New method for ultrasonic Doppler probe angle determination. AB - A new method is described to determine the Doppler probe angle using a standard pulsed Doppler; the technique is based on the determination of vessel position with respect to the skin surface. The transit times of ultrasound waves toward the vessel from multiple probe positions on the skin, the distance between probe positions, and the probe-skin angle are measured. Using the probe-skin angle and the slope of the regression curve, the vessel-skin angle is determined trigonometrically. Experimental results show high accuracy within a vessel-skin angle range of 5-40 degrees. PMID- 2969065 TI - Circulating concentrations of porcine ileal peptide but not hexosaminidase are elevated following 1 hr of mesenteric ischemia. AB - Porcine Ileal Peptide (PIP) is located in the mucosa of the small bowel. We hypothesized that PIP may be useful as a marker for early intestinal ischemia or other acute processes of the mucosa. To test this hypothesis we developed a model of acute reversible intestinal ischemia in the pig. Following isolation of a 100 cm segment of ileum on a vascular pedicle baseline, serum and tissue samples were obtained. The vessels were then occluded for 60 min and the segment was reperfused. Serial serum samples were taken and analyzed for PIP and hexosaminidase (HEX). HEX enzyme activity in serum is known to be elevated in animals having intestinal necrosis. The Student t test for paired data was used. In preliminary studies we found that circulating HEX activity became elevated following 3 to 4 hr of vessel occlusion followed by reperfusion. In the current experiments, following 1 hr of ischemia, PIP rose significantly in the peripheral circulation, being 153.8 +/- 76.8, 909.0 +/- 150.4, and 898.3 +/- 128.1 ng/ml (P less than 0.001) at 0, 60, and 360 min after reperfusion of the segment. HEX on the other hand did not change significantly throughout the experiment, having been 766.0 +/- 28.1, 752.0 +/- 71.3, and 780.1 +/- 53.7 nM/liter (ns) at 0, 60, and 360 min following reperfusion of the segment. Histology demonstrated some clubbing, shortening and fracturing of villi with thinning of the tips of the villi in many cases. Immunospecific staining for PIP was present along the intact borders of the villi.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969066 TI - [Knowledge of the truth in terminal patients]. PMID- 2969067 TI - [Report on the use of hepatitis B vaccine in health personnel in Spanish hospitals. The Spanish Group for the Study of Viral Hepatitis]. PMID- 2969068 TI - [Attitude of high-risk personnel to a hepatitis B vaccination campaign in a general hospital]. PMID- 2969069 TI - [Hepatitis B vaccination of health personnel]. PMID- 2969070 TI - Prospective study of cutaneous phototoxicity after systemic hematoporphyrin derivative. AB - Hematoporphyrin derivative (HpD) is a photoactive, oncophilic substance that produces cutaneous photosensitivity as its only significant side effect. Twenty three patients who received systemic HpD and the usual light-avoidance precautions were studied prospectively to determine the incidence and severity of cutaneous phototoxicity (CP). Seventeen of the 23 patients (74%) reported CP, including three patients (18%) who experienced blister formation. Symptoms of CP occurred for a mean duration of 6 weeks (range 5-23 weeks). Lack of compliance with restrictive photoprotective measures was felt to be a major contributing factor. Other HpD-related complications included skin hyperpigmentation, ocular discomfort, pruritus, pain at injection site, and urticaria. CP and the restrictive measures to avoid it represent major disadvantages of the clinical use of HpD. PMID- 2969071 TI - Photodynamic therapy: response of normal canine urethra using a cylindrical fiber. AB - Normal urethral response to photodynamic therapy (PDT) utilizing a cylindrical fiber was assessed in ten study and two control NIH fox-hounds. The canine urethras were treated 48 hours after intravenous injection of 3 mg/kg dihematoporphyrin ether (Photofrin II). A 1-mm fused silica optical fiber, with the distal 2-3 cm modified for cylindrical light distribution (660-microns diameter), was placed in the pendulous urethra. An argon-pumped dye laser delivered 100 mW of 630-nm light for 7 minutes (42 J) to the treatment area. Urethroscopy using either a 2.5-mm or a 2.8-mm flexible pediatric bronchoscope with video recording capability was performed immediately after PDT (12 animals) and 6 weeks later (nine animals); intravenous urography was performed 3 days and 6 weeks after PDT. There was no evidence of either urethral damage or stricture formation in any animal. Histopathology of the first three study specimens, obtained at 6 weeks, confirmed the absence of any pathology. Flexible instrumentation facilitates post-treatment assessment. This study demonstrates for the first time that, when a cylindrical fiber is used, photodynamic therapy may be applied to the urethra without damage to the normal urethral mucosa. This result has implications for potential treatment of initial or recurrent carcinoma in situ of the urethra using PDT. PMID- 2969072 TI - Laser angioplasty: an atherosclerotic swine model. AB - Rapid production of occlusive, atherosclerotic iliac artery lesions was achieved in 25 of 27 (93%) Yucatan miniature swine, using a combination of high cholesterol diet and mechanical endothelial denudation. Animals were fed a diet with 2% of their calories as raw cholesterol 2 weeks prior to balloon denudation of iliac arteries, which resulted in atherosclerotic lesions within 8 weeks. Early after denudation we have demonstrated total occlusion of arteries by fibrin thrombi, which in time organize and ultimately result in fibrotic occlusive disease. The arterial walls and intima show varying degrees of foam cell infiltration with destruction of the internal elastic lamina and calcification. Totally occluded lesions show fibrointimal proliferation, fibrosis, and multiluminal channels, which are probably secondary to organized thrombus. Our model of occlusive iliac artery disease involving vessels of 1 to 3 mm in diameter allows the development of catheter systems suitable for use in human peripheral and coronary arteries. This model is useful for the study of angioplasty, whether mechanical, balloon, or laser-mediated. PMID- 2969073 TI - The effect of porta-caval shunt, ammonia infusion and alcohol administration on rat plasma beta-hexosaminidase. AB - The activity of the lysosomal enzyme, beta-hexosaminidase, is increased in plasma of patients with various forms of liver disease as well as in plasma from rats with experimental cholestasis or cirrhosis. In this experimental study in the rat, the effect of porta-caval shunt, ammonia infusion and ethanol feeding on plasma beta-hexosaminidase activity was studied. Porta-caval shunted animals had significantly increased plasma beta-hexosaminidase activity compared to sham operated animals. Ammonia infusion in porta-caval shunted rats resulted in a further increase of plasma enzyme activity. Ethanol feeding for different periods of time (1 day to 4 weeks) did not have any influence on plasma beta hexosaminidase activity. PMID- 2969074 TI - ATPase activity in cells of the yeast Saccharomycopsis fibuligera during mycelial growth. AB - Levels of ATPase activity were examined in the cells of Saccharomycopsis fibuligera during mycelial growth. The mycelial cells (M-form cells) grown under standing culture conditions exhibited a marked increase of ATPase activity after 3 days, showing a broad peak between 5 and 20 days and a maximum at 10 days. The dry weight/ml of the yeast-like cells (Y-form cells) grown under shaking culture conditions reached its peak after 3 days and remained constant throughout the 10 day period of observation. The dry weight/ml of the M-form cells under standing culture conditions exhibited its peak in 10-15 days and then gradually decreased. A very small amount of Y-form cells among the M-form cells under standing culture conditions, exhibited very low ATPase activity even in 10 days' incubation. In M form cells, the particles increased in number during 20 days of incubation. PMID- 2969075 TI - Periodic cutaneous blood flow during aldehyde-provoked hyperemia. AB - Forearm cutaneous blood flow was monitored continuously by laser Doppler velocimetry in 12 normal human subjects before and after a 5-min topical challenge with 5 M propionaldehyde. The aldehyde challenge routinely provoked an increase in cutaneous blood flow. During the recovery phase from peak stimulated blood flow to a lower, stable, resting level of flow, the cutaneous blood flow exhibited rhythmic oscillatory activity. Three stages of oscillatory vasomotion were defined to characterize changes: the first 5 min after onset (I), the 5-min span bracketing the temporal midpoint (II), and the final 5 min (III). The average period, wave height, and erythrocyte flux decreased during these three stages of oscillatory vasomotion. These changes and the temporal characteristics of onset and disappearance of oscillatory vasomotion suggest an origin of the oscillations in the slow wave activity of vascular smooth muscle. PMID- 2969076 TI - Two-dimensional flow cytometric analysis of T cell subsets maintained in IL-2 medium after autologous mixed leukocyte reaction activation. AB - Human T cells are stimulated with an autologous mixed leukocyte reaction (AMLR) and can be propagated in interleukin-2. Staining of the cultured cells with the combination of two monoclonal antibodies was evaluated by two-dimensional flow cytometry at weekly intervals. AMLR activation resulted in an initial preservation of the CD4+ (helper/inducer T) subset predominance over the CD8+ (suppressor/cytotoxic T) cells, noted on normal circulating blood lymphocytes. However, during culture in interleukin-2, there was a progressive increase in the percentages of CD8+ Leu 15- cytotoxic T, CD4+ Leu 8- helper T, and CD3+ HLA-DR+ activated T cells, and a concomitant decrease in those of CD4+ Leu 8+ suppressor inducer T and CD8+ Leu 15+ suppressor T cells if the responder sheep red blood cell (SRBC)-rosetting T cells were made up by tris ammonium chloride, but not by hypotonic shock treatment to lyse SRBC. The significant difference between hypotonic shock-T cells and ammonium chloride-T cells in the phenotypic changes of T cell subsets after long-term culture in an interleukin-2 medium may suggest a regulatory role of the ammonium chloride-sensitive T cells in the AMLR. PMID- 2969077 TI - Progressive formation of DNA lesions during treatment with anti-metabolites without incorporation of the drugs into DNA. AB - Anti-metabolites, such as methotrexate, 5-fluoropyrimidines or hydroxyurea, induce progressive formation of DNA lesions. 5-Fluoropyrimidines induce DNA lesions either by incorporation of the drug into DNA or by a mechanism not involving incorporation. The second mechanism, not involving incorporation, is also seen with methotrexate and hydroxyurea. The three anti-metabolites have in common their ability to reduce intracellular levels of nucleotides, resulting in reduced efficiency of repair of DNA lesions. The lesions probably appear spontaneously, independently of the drug treatment. PMID- 2969078 TI - Deoxyribonucleoside triphosphate pools in Neurospora crassa: effects of histidine and hydroxyurea. AB - An effective HPLC method for detecting deoxyribonucleoside triphosphates in hyphae from the fungus Neurospora crassa has been developed. In rapidly growing cells the nucleotide levels vary from 11.8 pmoles/micrograms DNA for dGTP to 24.2 pmoles/micrograms DNA for dTTP. These levels fall by approximately one half in stationary-phase cultures but the ratio of each pool to dGTP remains the same. The dNTP pools in conidia are at least 5-fold lower than in rapidly growing cells. The pool sizes are the same in static and shaking cultures. When the ribonucleotide reductase inhibitor, hydroxyurea (30 mM), is added to rapidly growing cultures, DNA synthesis is stopped and the dGTP pool is reduced by 39%, while the size of the other pools remains the same. In the presence of 11 mM histidine, DNA synthesis is also stopped and the size of the dGTP pool reduced by 46% while the deoxypyrimidine pools are somewhat increased. This suggests that the toxicity of excess histidine in Neurospora may be due to its ability to interact with the ribonucleotide reductase, inactivating the enzyme. Histidine may react with the free radical at the active site, as does hydroxyurea. PMID- 2969079 TI - Coronary angioplasty 1985-1986 and 1977-1981. PMID- 2969080 TI - Back pain and sciatica. PMID- 2969083 TI - 8-Hydroxy-2-(di-n-propylamino) tetralin, a selective serotonin1A receptor agonist, blocks haloperidol-induced catalepsy by an action on raphe nuclei medianus and dorsalis. AB - The selective serotonin1A receptor agonist 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) was studied for its ability to reverse haloperidol-induced catalepsy in rats. Given subcutaneously 8-OH-DPAT (0.06-0.5 mg/kg), dose dependently antagonized the catalepsy induced by 1 mg/kg of haloperidol. Intraventricular injection of the serotonin (5-HT) neurotoxin 5,7 dihydroxytryptamine (5,7-DHT), which caused marked depletion of 5-HT in brain, did not change haloperidol-induced catalepsy per se, but completely antagonized the anticataleptic effect of subcutaneously administered 8-OH-DPAT. When injected directly into the median or dorsal raphe nucleus, 8-OH-DPAT, in doses ranging from 0.2 to 5 micrograms/0.5 microliter, reduced the catalepsy induced by haloperidol. The results suggest that the activation of 5-HT1A receptors, probably those located presynaptically on 5-HT-containing cell bodies, reduces the catalepsy induced by haloperidol. PMID- 2969082 TI - The effect of pretreatment with pentobarbital on the extent of [14C] incorporation from [U-14C]glucose into various rat brain glycolytic intermediates: relevance to regulation at hexokinase and phosphofructokinase. AB - In the present investigation we monitored the incorporation of [14C] from [U 14C]glucose into various rat brain glycolytic intermediates of conscious and pentobarbital-anesthetized animals. Labeled glucose was delivered to brain by single bolus intracarotid injection and brain tissue was subsequently prepared at 15, 30, and 45 sec by freeze-blowing. Glycolytic intermediates were then separated by column chromatography. Our results showed a gradual decrease with time of 14C-labeled glucose which gave a calculated rate for glucose metabolism of 0.86 mumol/min/g and 0.56 mumol/min/g in conscious and anesthetized animals, respectively. Compared to the results obtained using conscious animals the administration of pentobarbital not only resulted in a significant attenuation of the rate of glucose metabolism but also caused a similar reduction in the amount of 14C incorporated into several glycolytic intermediates. These intermediates included: glucose 6-phosphate, fructose 6-phosphate, fructose 1,6 diphosphate, dihydroxyacetone phosphate and post glycolytic compounds. In addition, pretreatment with pentobarbital resulted in a 75% increase in the endogenous concentration of glucose, 10% increase in glucose 6-phosphate, no change in fructose 6-phosphate and 42% decrease in lactate compared to levels in brains obtained from conscious animals. These results are discussed in relation to control of glycolysis through coupled regulation at hexokinase phosphofructokinase. PMID- 2969081 TI - Heparin and heparinoids impair adrenaline and platelet-activating factor but not thrombin-induced inhibition of adenylate cyclase and stimulation of GTP hydrolysis in human platelet membranes. AB - The effects of heparins and heparinoids were studied on adenylate cyclase and GTPase activities in human platelet membranes. Inhibition of adenylate cyclase by adrenaline and platelet activating factor was completely abolished by heparin at 1 microgram/ml. At similar concentration heparin blocked the stimulation of high affinity GTPase(s) by these hormonal factors. In contrast, heparin (up to 30 micrograms/ml) did not abolish adenylate cyclase inhibition and stimulation of GTP hydrolysis by thrombin in the absence of antithrombin III. In the presence of antithrombin III, thrombin action on adenylate cyclase was blocked by unfractionated and high molecular weight heparin at 0.1 microgram/ml. Low molecular weight heparins and pentosanpolysulfate were less or not effective. In contrast, all high and low molecular weight heparins tested were almost equally potent in inhibiting adrenaline-induced inhibition of adenylate cyclase in the absence of antithrombin III. The data indicate that heparins discriminate platelet activating factor and adrenaline-induced inhibition of adenylate cyclase from the inhibitory action of thrombin and delineate different structural requirements for the interaction of heparins with the adenylate cyclase system and antithrombin III. PMID- 2969084 TI - Fetal echocardiographic signs of congenital endocardial fibroelastosis. AB - Endocardial fibroelastosis is characterized by a porcelain-like thickening of the endocardium, resulting in a marked increase in echodensity of the endocardium, as well as ventricular dilatation and aortic atresia. With improvement in prenatal ultrasound, this condition can be suspected in utero on the basis of ventricular enlargement, poor ventricular contractility, and marked echodensity of the endocardial surface. We present two cases in which such conditions were found on prenatal M-mode echocardiography and two-dimensional directed pulsed Doppler. Ventriculomegaly and hypocontractility of the ventricle are, however, nonspecific for such conditions; the diagnosis can be made accurately only by pathology. When such findings appear on ultrasound, all efforts should be made to deliver the patient in a perinatal center for optimal neonatal surgery to improve the survival of the newborn. PMID- 2969086 TI - Periodontal disease and AIDS. PMID- 2969085 TI - Ultrasound-guided transvaginal needle aspiration of follicles for in vitro fertilization. AB - In search of a simpler and less traumatic method of oocyte retrieval for in vitro fertilization (IVF), 37 infertile patients underwent ultrasound-guided transvaginal needle aspiration of the follicles instead of the conventional laparoscopic aspiration. Although only intravenous analgesics and sedatives were used, patients experienced little discomfort. There were no infections or bleeding complications after the procedure. Eleven patients (30%) achieved ongoing pregnancies; this rate of success is comparable to that with our laparoscopic method. Considering this reasonable pregnancy rate, lack of substantial morbidity, and good patient acceptance, laparoscopic oocyte retrieval for IVF can be largely replaced by the ultrasound-guided transvaginal needle aspiration technique. PMID- 2969087 TI - Avian pulmonary hypertension syndrome. IV. Increased right ventricular mass in turkeys experimentally infected with Plasmodium durae. AB - Infection of young turkeys with Plasmodium durae produced a significant increase in the mean relative right ventricular mass from 0.172 in the controls to 0.208 in the infected group. Although turkeys react less strongly to hypoxia than broiler chickens, the similarity of this finding to that previously reported in broilers infected with Aegyptianella pullorum suggests that the hypoxic effect of the anaemia produced by these blood parasites causes the pulmonary hypertension and subsequent right ventricular hypertrophy. PMID- 2969088 TI - Professional liability #5. The role of the consulting examiner in workmen's compensation examinations. PMID- 2969089 TI - Constant inclined pelvic traction for treatment of low back pain. AB - The authors describe a technique of continuous pelvic traction applied to a select group of 46 patients with low back pain, who had not responded to at least two weeks of treatment by bed rest and analgesic medication before hospitalization. The traction technique included the use of force equivalent to 25% of body weight, directed at an angle of (15) to the plane of the bed. A mechanical analysis of the technique is described and the results of treatment and the efficacy of the technique discussed. PMID- 2969090 TI - Role of consulting examiner in workmen's compensation. PMID- 2969091 TI - Thiamin status of the offspring of diabetic rats. AB - Erythrocyte transketolase activity and thiamin pyrophosphate effect were examined in the offspring of streptozotocin-diabetic rats. Thiamin reserve was found to be significantly reduced in litters of untreated diabetic rats as compared to control and to insulin-treated diabetic rats. Supplementation of the untreated diabetic dams throughout pregnancy with oral thiamin was associated with a significantly improved thiamin status of the litters. We conclude that, due to enhanced fetal glucose turnover during diabetic gestation, a fetal thiamin deficiency state may evolve; this condition can be remedied with maternal thiamin supplementation. PMID- 2969093 TI - Caring around the clock. PMID- 2969092 TI - Regional infusion of fluoropyrimidines for hepatic metastases of colorectal cancer. AB - Recent reports on regional chemotherapy for hepatic metastases of colorectal origin are reviewed. Because of reported side effects and lack of evidence for its superiority over conventional intravenous administration, intra-arterial administration of fluoropyrimidines cannot be advised for routine use, but should be restricted to centres participating in clinical trials on this subject. PMID- 2969094 TI - [Vasodilators for intracoronary use]. PMID- 2969095 TI - [Comparison of intravenous thrombolysis and intracoronary thrombolysis]. AB - Any comparison between intravenous and intracoronary thrombolysis must take into account such elements as the delays imposed by coronary arteriography, the extra cost of the procedure, etc., but ultimately the answer can only be provided by a controlled trial comparing the two approaches. To give truly significant results, this trial should fulfill several requirements concerning the time of randomization to treatments, the procedure to be followed in each group and the criteria of comparison. No such trial is available at present, so that the comparison can only rest on speculations and less demonstrative facts. As it stands, it is not in favour of intracoronary thrombolysis. PMID- 2969096 TI - Coronary angioplasty for unstable angina. AB - The clinical syndrome of unstable angina causes great concern to clinicians because of the high risk of progression to myocardial infarction or cardiac death. Management of unstable angina pectoris has evolved progressively, and recently coronary angioplasty has been shown to be a relatively safe and effective treatment of unstable angina. The aim of this study was to report our experience with the immediate and one year follow-up results of percutaneous transluminal coronary angioplasty (PTCA) in patients with unstable angina. Unstable angina was defined as prolonged periods of chest pain at rest, associated with ST-T changes and no subsequent myocardial necrosis. All patients were treated with a combination of nitroglycerin, beta-blockers or calcium antagonists. The patients were classified into three clinically relevant subgroups. Group I: unstable angina stabilized with pharmacological treatment but with persistent exertional angina (71 patients): group II: unstable angina refractory to optimal pharmacological treatment (88 patients) and group III: recurrent unstable angina within 30 days of myocardial infarction (53 patients). The initial success rate was 87 p. 100 for group I, 92 p. 100 for group II and 89 p. 100 for group III and the major complication rates were 13 p. 100, 8 p. 100 and 11 p. 100 respectively. At 1 year follow-up after initial successful PTCA late death had occurred in 2 p. 100 in group I, 1 p. 100 in group II and 0 p. 100 in group III and a late nonfatal infarction in 2 p. 100, 1 p. 100 and 4 p. 100 respectively. The frequency of recurrent angina was 23 p. 100 in group I, 20 p. 100 in group II and 26 p. 100 in group III. Thus, coronary angioplasty for unstable angina can be performed with a high initial success rate, but at an increased risk on major complications. The prognosis after an initial successful coronary angioplasty is favourable. PMID- 2969097 TI - [Intracoronary treatments in emergency situations]. AB - Mechanical and medicinal therapies are widely used in coronary disease patients for emergency or semi-emergency situations. Vasodilator drugs injected into arteries that are occluded or narrowed-drugs injected into arteries that are occluded or narrowed by spasm rapidly relieve that spasm; injecting these drugs prior to angioplasty renders the myocardium more tolerant to the ischaemia created by the dilated balloon. In unstable angina coronary angioplasty has a 90 p. 100 primary success rate with immediate complications that are slightly more frequent than in stable angina; compared to historical series with medicinal treatment, angioplasty reduces the risk of myocardial infarction and/or secondary death. In evolving myocardial infarcts the respective values of intracoronary thrombolysis and angioplasty are still under discussion, although the results of recent studies tend to favour intravenous thrombolysis and secondary, elective angioplasty. PMID- 2969098 TI - [SIN-1 in intracoronary administration]. AB - SIN-1, a new anti-anginal compound, has been the object of extensive clinical and experimental research work. It has been injected into the coronary arteries in more than 1,000 patients, enabling its effects on coronary circulation to be studied without interference from its systemic effects. Contrary to dipyridamole or calcium channel blockers, SIN-1 dilates the main epicardial coronary vessels without blood steal. The dilatation it induces is rapid and prolonged, and its intensity is at least equivalent to that of nitroglycerin, nitrites and calcium inhibitors. Its action lasts longer than that of nitroglycerin. SIN-1 enables balloon inflation to be prolonged in percutaneous transluminal coronary angioplasty. It is effective in relieving coronary spasm and has a favourable effect on anterograde and collateral circulation. Haemodynamically, SIN-1 significantly reduces preload but has little effect on afterload; it does not alter left ventricular contractility and has no influence on peripheral resistance. Owing to its rapid and long-lasting dilating effect specifically on the epicardial coronary arteries, its effectiveness against coronary spasm and its lack of side-effects, SIN-1 is indicated for coronary arteriography and coronary spasms. PMID- 2969099 TI - [Anti-ischemic effects of SIN-1, a molsidomine metabolite, during coronary angioplasty and antiplatelet effects in humans]. AB - The effects of SIN-1, the biologically active metabolite of molsidomine, on tolerance of ischaemia during percutaneous transluminal coronary angioplasty were studied in 12 patients. Following an intracoronary injection of SIN-1 0.5 mg, the time required for the ST segment to be depressed by at least 0.2 mV was significantly prolonged from 44 to 55-62 seconds. Simultaneously, SIN-1 reduced the significant rise in left ventricular end-diastolic pressure observed during dilatation. One minute after SIN-1 was injected, no effect was noted or on the increase in left ventricular volume indices or on the reduction of ejection fraction associated with dilatation. The maximum anti-ischaemic effect of SIN-1 was obtained 5 minutes after the injection. As regards the mechanism of action of the drug, a decrease in parietal tension through reduction of left ventricular preload should be considered. In addition, strong inhibition of platelet aggregation and adenosine triphosphate secretion was demonstrated in high platelet content plasma and whole blood 2 hours after an oral 16 mg dose of molsidomine. The antiplatelet effect of molsidomine may contribute to its anti ischaemic effect by reducing both platelet adherence at the site of dilatation and the release of vasoconstrictor and pro-aggregant mediators. PMID- 2969100 TI - [Role of molsidomine on platelet activation in coronary ischemia]. AB - Platelet suppressive agents have been shown to improve the prognosis of coronary diseases such as myocardial infarction and unstable angina. Several markers of platelet activation during myocardial ischemia have been found to be increased. Platelet granule constituents (beta thromboglobulin or platelet factor 4) or thromboxane B2 have been reported to be enhanced and, in some studies, to be correlated with the ischemia. Molsidomine or its active metabolite SIN-1 have antithrombotic properties in experimental models. This effect seems to be at least partly related to their antiplatelet activities. SIN-1A inhibited platelet aggregation and release reaction. Specific investigations have demonstrated that SIN-1A acts at a early stage of platelet activation inhibiting calcium influx and phospholipase activity which lead to inhibition of thromboxane formation and fibrinogen binding. Antiplatelet properties were also observed after oral administration of molsidomine but the extent of inhibition appeared to vary with the subjects. PMID- 2969101 TI - [The family of sydnonimines (Corvasal, intracoronary Corvasal 1 mg). Analogies with EDRF (endothelium-derived relaxing factor)]. PMID- 2969102 TI - [Coronary insufficiency. Biological and clinical update. The value of sydnonimines. International symposium. Lyons, 2-3 October 1987. Proceedings]. PMID- 2969103 TI - [Pharmacological bases of therapeutic tolerance]. AB - The purpose of this review paper is to describe the various mechanisms that may lead to a decrease in the effectiveness of drugs during long-term treatment (the so-called "tolerance" or "escape phenomenon"). In addition, some precise recommendations will be made. Tolerance may be due either to degradation of the active substance by enzyme induction, as is the case e.g. with barbiturates (pharmacokinetic drug tolerance) or to down-regulation of receptors (e.g. beta adrenergic drugs and opiates) or exhaustion of the metabolic pathways involved in pharmacological activation, as with nitrates and other drugs (pharmacodynamic drug tolerance). Whatever its mechanism, tolerance can be prevented by intermittent drug administration despite the problems inherent in the therapeutic gaps thus created. Another, more rational approach is to replace the drug concerned by a more appropriate drug which permits, or induces the restoration of receptor density when their number is reduced by the physiopathological situation (e.g. H2 stimulation instead of beta-adrenergic stimulation) or which reproduces the action of first physiological messengers, such as the endothelium-derived relaxing factor (EDRF), without requiring metabolic activation (e.g. SIN-1 or molsidomine instead of nitrates). PMID- 2969105 TI - [The tolerance phenomenon in the therapeutic management of angina pectoris]. AB - Tachyphylaxis in the treatment of angina pectoris may be found in widely different situations, such as stable angina turned unstable owing to local coronary changes, interference with another pathology (intricate angina) or true "escape phenomenon". In clinical practice, angina from tachyphylaxis may be due to a variety of causes, including lack of compliance (often difficult to diagnose), untimely drug administrations, insufficient dosage or habituation. Habituation corresponds to adaptation of the cells to their chemical environment. In the case of angina, it is due to gradual depletion of sulfhydryl radicals in the vascular endothelial cells. This progressive insensitivity to nitrites accounts for some therapeutic failures. Molsidomine is thought to be of great interest in such cases. PMID- 2969104 TI - [Comparative course of maintenance of the effects of nitrite derivatives and molsidomine after prolonged administration]. AB - Prolonged administration of nitroglycerin and its derivatives results in pharmacodynamic tolerance: although the dosage of these drugs is kept at the same level, their therapeutic effects decrease in amplitude. Experimental and clinical studies have shown that this escape phenomenon is due to depletion of cysteine in the vascular wall. This amino acid, which gives off SH radicals, is indispensable to the ultimate transformation of nitroglycerin enabling in to exert its vasodilator action. Molsidomine does not require any transformation to act on the vascular smooth muscle and should therefore remain insensitive to tolerance. The experimental and clinical data available at present seem to confirm that the effects of molsidomine administered for long periods in the treatment of coronary disease and heart failure are sustained. However, for the long-term effectiveness of molsidomine and nitrites to be compared objectively a prospective double-blind randomized trial would be needed, with both treatments being in optimal doses and intervals of administration. PMID- 2969107 TI - What all dental personnel should know about AIDS. PMID- 2969106 TI - [Immunological characteristics of patients with diffuse toxic goiter]. AB - Altogether 24 patients with diffuse toxic goiter were examined after surgical intervention and thiamazole therapy. In addition to clinical methods the levels of T- and B-lymphocytes, immunoregulatory cells (T-helpers and T-suppressors) and immunoglobulins of different classes were determined before and after therapy. A method of spontaneous and complementary rosette formation and monoclonal antibodies (Ortho) were used for the determination of the content of T- and B lymphocytes. Immunoglobulins were determined by a method of radial immunodiffusion in gel. It was shown that in patients with diffuse toxic goiter the level of T-lymphocytes was significantly on a decrease, the levels of B lymphocytes and IgG were on an increase before therapy. There was sharp disturbance in the ratio of immunoregulatory cells--T-helpers and T-suppressors. After therapy the clinical picture and immunological indices in 14 patients returned to normal, in the rest of 10 patients the Ig level and ratio of immunoregulatory cells remained disturbed. PMID- 2969108 TI - Efficacy of a cell-mediated reaction to the purified protein derivative of tuberculin in the disposal of Mycobacterium leprae from human skin. AB - The purpose of this study was to evaluate the effects of a delayed-type cell mediated immune response to Mycobacterium tuberculosis antigen on the Mycobacterium leprae load in the skin of leprosy patients. Twelve patients with the lepromatous form of leprosy have been injected intradermally with 5 units of the purified protein derivative of tuberculin (PPD). Ten individuals responded with areas of induration ranging from 12 to 21 mm in diameter, and two were unresponsive (less than 10 mm). Twenty-one days thereafter, the injected and control sites were biopsied, and the histology, number of acid-fast bacilli, nature and phenotype of the emigrant cells, and ultrastructural characteristics of the lesions were evaluated. Eight of the 10 responding patients showed reductions in the number of acid-fast bacilli by factors ranging from 5 to 10,000. Two responders and both nonresponders exhibited no discernible decline in the number of organisms. The reduction in bacillary load was correlated with an intense mononuclear cell infiltrate, the maintenance of a high CD4+ T-cell/CD8+ T cell ratio, the formation of granulomata, and the extensive destruction of previously parasitized macrophages. PMID- 2969110 TI - Female rats are susceptible to cardiac hypertrophy induced by copper deficiency: the lack of influence of estrogen and testosterone. AB - In contrast to a previous report (Fields M, Lewis C, Scholfield DJ, Powell AS, Rose AJ, Reiser S, Smith, JC. Proc Soc Exp Biol Med 183:145-149, 1986), female rats were shown to be susceptible to copper (Cu) deficiency giving rise to restriction of growth, cardiac hypertrophy, and anemia. The severity of these effects was, however, found to be less marked than in the male rats which had similar liver Cu levels. Castration or ovariectomy of Cu-deficient rats had little effect on CH or the other parameters associated with Cu deficiency, and supplementation of the neutered animals with estrogen or testosterone was similarly without effect. The ultrastructural appearance of the hypertrophied Cu deficient female heart was similar to that previously found in males and was characterized by a large increase in mitochondrial area with disrupted cristae. The results also indicated that in contrast to Cu-deficient males iron (Fe) was not accumulated in the liver of the Cu-deficient female rats. It may be concluded that the limited protection of female rats to the effects of Cu deficiency observed in this study were unconnected with the sex steroids. PMID- 2969109 TI - Tissue-specific expression and cDNA cloning of small nuclear ribonucleoprotein associated polypeptide N. AB - Sera from some patients with systemic lupus erythematosus and other autoimmune diseases have antibodies against nuclear antigens. An example is anti-Sm sera, which recognize proteins associated with small nuclear RNA molecules [small nuclear ribonucleoprotein (snRNP) particles]. In this paper anti-Sm sera were used to probe immunoblots of various rat tissues. A previously unidentified Mr 28,000 polypeptide was recognized by these anti-Sm sera. This polypeptide, referred to as "N," is expressed in a tissue-specific manner, being most abundant in rat brain, less so in heart, and undetectable in the other tissues examined. Immunoprecipitation experiments using antibodies directed against the cap structure of small nuclear RNAs have demonstrated that N is a snRNP-associated polypeptide. Anti-Sm serum was also used to isolate a partial cDNA clone (lambda rb91) from a rat brain phage lambda gt11 cDNA expression library. On RNA blots, the 450-base-pair cDNA insert of this clone hybridized to a 1600-nucleotide mRNA species with an identical tissue distribution to N, suggesting that lambda rb91 encodes at least part of N. A longer cDNA clone was obtained by rescreening the library with lambda rb91. In vitro transcription and subsequent translation of this subcloned, longer insert (pGMA2) resulted in a protein product with the same electrophoretic and immunological properties as N, confirming that pGMA2 encodes N. The tissue distribution of N and the involvement of snRNP particles in nuclear pre-mRNA processing may imply a role for N in tissue-specific pre-mRNA splicing. PMID- 2969111 TI - Alterations in lipid composition and fluidity of liver plasma membranes in copper deficient rats. AB - In view of the importance of membrane fluidity on cell functions, the influence of phospholipid acyl groups on membrane fluidity, and the changes in lipid metabolism induced by copper (Cu) deficiency, this study was designed to examine the influence of dietary Cu on the lipid composition and fluidity of liver plasma membranes. Male Sprague-Dawley rats were divided into two dietary treatments, namely Cu deficient and Cu adequate. After 8 weeks of treatment, liver plasma membranes were isolated by sucrose density gradient centrifugation. The lipid fluidity of plasma membranes, as assessed by the intramolecular eximer fluorescence of 1,3-di(1-pyrenyl) propane, was significantly depressed by Cu deficiency. In addition, Cu deficiency significantly reduced the content of arachidonic and palmitoleic acids but increased the docosatetraenoic and docosahexaenoic acids of membrane phospholipids. This alteration in unsaturated phospholipid fatty acid composition, especially the large reduction in arachidonic acid, may have contributed to the depressed membrane fluidity. Furthermore, Cu deficiency also markedly altered the fatty acid composition of the triacylglycerols associated with the plasma membranes. Thus, the lipid composition and fluidity of liver plasma membranes are responsive to the animal's Cu status. PMID- 2969112 TI - Melatonin acts in the brain to mediate seasonal steroid inhibition of luteinizing hormone secretion in the white-footed mouse (Peromyscus leucopus). AB - A melatonin-induced supersensitivity of the gonadotropin-secretory system to the negative feedback action of sex steroids is thought to be important to the timing of seasonal reproduction. However, little is known concerning this action of melatonin. In the present study the antigonadal action of melatonin in the anterior hypothalamus (AH) of the white-footed mouse, Peromyscus leucopus, was used to examine the neuroendocrine mechanism whereby melatonin enhances the sensitivity to sex steroid negative feedback. Mice received a melatonin containing pellet in the AH for 14 weeks, at which time they were castrated and treated sc with a Silastic testosterone (T) capsule for 3 weeks. At the time of castration, weight of the testes and the concentration of T in the blood of mice with a melatonin pellet were greatly reduced compared to mice with a blank (melatonin-free) implant in the AH (P less than 0.01). In mice treated with melatonin the physiological dose of T significantly reduced the concentrations of LH in blood and pituitary (P less than 0.05). This dose of T, however, had little effect on LH in mice with a blank pellet in the AH. Melatonin in the AH markedly increased the content of gonadotropin-releasing hormone (GnRH) in the mediobasal hypothalamus (P less than 0.05) in mice treated with T; however, there was little effect of melatonin and/or T in any other region examined. Melatonin and T had little effect on the contents of immunoreactive beta-endorphin (B-EP) in the hypothalamus, but T alone increased the content of B-EP in the preoptic area. These results are evidence that melatonin and T act in concert to induce the reproductively-quiescent state by suppressing secretion of GnRH from the hypothalamus. PMID- 2969113 TI - A high-salt meal produces natriuresis in humans without elevating plasma atriopeptin. AB - The effects of a high-sodium meal on plasma atrial natriuretic peptide (atriopeptin) and renal sodium excretion were studied in eight normal human subjects. As expected, sodium excretion and urine osmolality increased following the meal. Plasma atriopeptin levels did not increase, however, after the high sodium meal. In a control experiment, consumption of a low-sodium meal by six of the same subjects did not increase either urinary sodium excretion or plasma atriopeptin concentration. We conclude that the natriuresis elicited by a high salt meal is not mediated by the atrial peptides. PMID- 2969114 TI - Is there a best endocrine management of prostatic carcinoma? PMID- 2969115 TI - Needle endoscopy in special procedures. AB - A needle endoscope, with a rigid shaft 2.2 mm wide and 20 cm long, an illumination range of 2-4 cm, and excellent optics, has proved to be very valuable in performing special interventional procedures because it can be inserted in any relatively straight 7-F catheter track. The needle endoscope has been very useful in combination with fluoroscopy for retracing lost drain tracks, evaluating fistulas and sinuses, performing limited pleuroscopy and laparoscopy for biopsy under direct vision, and evaluating the gallbladder and renal pelvis for location of duct openings. The author describes techniques used with the needle endoscope and results in a sample of cases. There has been no morbidity associated with the use of the instrument in over 800 examinations in a 12-year period. PMID- 2969116 TI - Direct argon laser exposure for recanalization of peripheral arteries: early results. AB - Direct laser light, in combination with percutaneous transluminal angioplasty (PTA), was used to recanalize atherosclerotic peripheral arteries. Argon laser energy was controlled with an optical assembly and aligned with a special centering/dilation balloon so that plaque tissue absorbed laser energy directly and was vaporized. A channel was thereby created with multiple 10-W laser exposures (2-10 seconds), and conventional angioplasty was then performed. Recanalization was achieved in 33 of 36 procedures (92%), in 23 femoropopliteal (mean length, 9 cm) and three iliac total occlusions and ten femoropopliteal high grade stenoses, with the three failures occurring in the first group. Complications included two emboli, six moderate groin hematomas, and one laser perforation. While these results are preliminary (mean follow-up, 3 months), direct laser angioplasty appears to be a useful adjunct to PTA for treating atherosclerotic arteries. PMID- 2969117 TI - Axillary and subclavian vein stenosis: percutaneous angioplasty. AB - Twenty-nine percutaneous balloon dilations of the axillary and subclavian veins were performed in 19 patients. Stenoses occurred in typical locations of anatomic narrowing or at sites of previous trauma. The initial success rate was 76%, with a 1-year patency rate of 35% and a 2-year patency rate of 6%. Angioplasty can be performed on an outpatient basis with a very low rate of significant complications and can be repeated numerous times to keep a vein patent for many years. This procedure is especially valuable in dialysis patients who have limited access sites. PMID- 2969118 TI - Epidural steroid injection: a procedure ideally performed with fluoroscopic control. AB - Epidural injection of steroid and local anesthesia can be used to treat low back pain. The injection is best performed with fluoroscopic control, with needle placement documented by means of a limited epidurogram. The technique was used in 116 patients; there were only three failures (2.5%) and one complication. PMID- 2969119 TI - [Luciferase]. PMID- 2969120 TI - Give a little of yourself to those who need help. PMID- 2969121 TI - [A simple procedure for the repair of the abdominal wall in the prevention and treatment of postoperative evisceration]. PMID- 2969122 TI - [Renal hydatid cyst]. PMID- 2969124 TI - [Gangrene of the extremities--terminology, classification and treatment. Our experience with 250 cases]. PMID- 2969123 TI - [Renal hydatid cyst]. PMID- 2969125 TI - [Etiopathogenetic and therapeutic considerations in traumatic colorectal lesions]. PMID- 2969126 TI - [Internal sphincterotomy in the treatment of anal fissures and hemorrhoids]. PMID- 2969127 TI - [2 cases of acute necrotizing fasciitis of the upper extremities]. PMID- 2969128 TI - [Multiple reinterventions following vagotomy combined with pyloroplasty in duodenal ulcer]. PMID- 2969129 TI - [Hypnoanalgesia by electrical hyperstimulation at acupuncture points as an anesthetic method in open-heart surgery]. PMID- 2969130 TI - [Gastric volvulus: an original treatment by reduction gastroplication and bipolar gastropexy]. PMID- 2969131 TI - Echocardiographic detection of pulmonary hypertension in anesthetized rats. AB - Evaluation of pulmonary arterial pressure in small laboratory animals requires surgical catheter implantation. However, even this modest surgical procedure is known to retard weight gain, alter various biochemical parameters and may exert other deleterious actions which could complicate data interpretation. To obviate these adverse effects of surgery and to permit better staging of experimental procedures, a method for non-invasive detection of pulmonary hypertension would be desirable. The present study examined the utility of m-mode echocardiography to detect increased pulmonary arterial pressure and right ventricular hypertrophy in rats with monocrotaline-induced pulmonary hypertension. At various times after administration of monocrotaline (105 mg/kg), rats were anesthetized lightly with sodium pentobarbital (30 mg/kg) and m-mode echocardiograms were generated using a 7 mHz transducer with the animals in the supine position. Mean pulmonary arterial pressure was then determined by closed-chest catheterization of the pulmonary artery. At sacrifice, the extent of right ventricular hypertrophy was determined as the ratio of the weight of the right ventricular free wall to that of the left ventricle plus septum. Right ventricular systolic time intervals, determined from the echocardiogram, correlated significantly with the extent of right ventricular hypertrophy (p = 0.05; r2 = 0.39) but not with mean pulmonary arterial pressure (p = 0.2; r2 = 0.14). Echocardiography may provide a simple, non-invasive means to detect cardiopulmonary abnormalities in rats and may thus be useful in staging subsequent experimental procedures without need for prior surgical catheter implantation. PMID- 2969132 TI - [Strangulated spigelian hernia]. PMID- 2969133 TI - Circadian-based infusional chrono-chemotherapy controls progressive metastatic renal cell carcinoma. AB - We treated 25 patients with progressive metastatic renal cell carcinoma with continuous infusion of 5-fluoro-2-deoxyuridine (FUDR) by implanted pump. FUDR was infused for 14 days at monthly intervals. Starting dose was 0.15 mg/kg/day intravenous or 0.25 mg/kg/day intra-arterial; intravenous doses were increased or decreased in increments of 0.025 mg/kg/day as permitted by toxicity. Circadian time modification of the infusion shape (sinusoidal with the peak centered around 6 p.m.) significantly lowered serious intravenous infusion-associated toxicity, allowing higher dose intensity. In 24 evaluable patients, two complete responses (8%), six partial responses (25%), and one minor response were seen. One secondary partial response was observed after infusional velban (total objective response rate 37.5%). Previously progressive disease was controlled in greater than 80% of our patients. During a median follow-up period of 8 months (range of 2-26 months), the overall survival for all 25 patients is 75%. Our results indicate that metastatic renal cell cancer responds to infusional chemotherapy and that the circadian shape of infusion markedly affects our ability to deliver effective doses. PMID- 2969135 TI - Infection control hotline. PMID- 2969136 TI - Do suppressor T cells exist? A reply. PMID- 2969134 TI - [The combined effect of an ethinylestradiol/desogestrel contraceptive combination on blood lipids and parameters of liver function and coagulation]. AB - The present study aims to evaluate the contraceptive efficacy of the new combination ethinylestradiol (30 micrograms)/desogestrel (150 micrograms) and, moreover, to demonstrate its low side effects compared to the symptoms present in the pretreatment cycles. In particular the authors took into consideration the changes in serum lipids, and hepatic and coagulation function parameters induced by pill, in 40 healthy women. An interesting result is that the estrogen induced increase of cholesterol-HDL is not antagonized by the desogestrel that has a slight affinity to androgenic receptors and SHBG. This is obviously of great importance in the prevention of atheromatous lesions. PMID- 2969137 TI - But still it moves! An answer to Professor Goran Moller. PMID- 2969138 TI - Suppressor versus cytolytic CD8+ T lymphocytes: where are the artefacts? PMID- 2969139 TI - Cytolysis by cloned helper T cells: induction by specific antigen or by anti-CD3 hybrid antibodies. AB - We have explored the use of hybrid antibodies--prepared by covalently linking anti-CD3 to an antibody specific for a monomorphic major histocompatibility complex (MHC) class II determinant using N-succinimidyl 3-(2 pyridyldithio)proprionate/succinimidyl 4-(N-malcimidomethyl)cyclohexane-1 carboxylate (SPDP/SMCC) as coupling reagent--in inducing cytolysis in human tuberculin (PPD)-specific T helper (TH) clones. These clones have been shown to lyse PPD-bound Epstein-Barr virus (EBV)-transformed B-cell lines (B-EBV) in an MHC class II-restricted manner. In this paper anti-CD4-induced cytolysis is compared with antigen/MHC-induced cytolysis with the same clones. Cytolysis induced by the hybrid antibodies was highly efficient, with killing of both syngeneic and allogeneic tumour cells positive for MHC class II. Conjugate induced cytolysis was maxima within 4 h; that of antigen-positive targets at 16 h. Killing of bystander cells was seen only when cytolysis was triggered by antigen/MHC, suggesting that the mechanism of cytolysis in the two systems may be distinct. Targets treated simultaneously with hybrid antibody and with antigen, thereby providing both activation signals to the clones, are lysed less efficiently than those treated with either PPD or hybrid antibody alone. Evidence is presented showing that this inhibition is most marked against syngeneic PPD coated cells treated with hybrid antibody, suggesting that two signals independently capable of activating cytolytic function in the clones, when presented simultaneously, interfere with the induction of the cytolytic process. PMID- 2969140 TI - Heterogeneity in the specificities of an alloreactive T helper cell line. AB - Heterogeneity in antigen recognition of an alloreactive helper T-cell line was studied by repeated limiting dilution cloning. Analysis of the fine specificities of these clones showed that the gain of reactivity to new antigens by the T-cell line included the generation of non-cross-reactive T cells specific to new major histocompatibility (MHC) antigens. Analysis of function revealed that all the T cell clones studied had similar cell surface antigens and secreted interleukin 4 (IL-4) and IL-5/IL-3 but not IL-2 or gamma interferon (IFN-gamma). The T-cell clones served as helper cells for B cells expressing the appropriate MHC antigens to both clonal proliferation and differentiation into antibody-producing cells. PMID- 2969141 TI - Enhancement of human spontaneous cell-mediated cytotoxicity by a monoclonal antibody against the large sialoglycoprotein (CD 43) on peripheral blood lymphocytes. AB - A murine monoclonal antibody, MoAb B1B6 (IgG1 chi), which recognizes the large sialoglycoprotein (LSGP) on human peripheral blood lymphocytes (PBL) effectively enhanced the spontaneous cytotoxicity of these cells against the natural killer (NK)-sensitive target cells K562 and Molt-4. Whereas preincubation of the lymphocytes with MoAb B1B6 resulted in increased cytotoxicity, preincubation of the target cells had no effect, indicating that the MoAb amplified cytotoxicity at the effector cell level. Kinetic analysis of the data revealed no differences between the control and the MoAb-treated lymphocytes with regard to Vmax, usually considered to reflect the overall lytic potential of the cells. The slopes of the saturation curves, however, differed significantly for the two cell populations, indicating a substantial increment in the activity of the MoAb-treated cells. When studied at the single cell level and with K562 as targets, treatment of PBL with the MoAb resulted in the recruitment of new effector lymphocytes from the pool of non-binding cells. In contrast, when Molt-4 cells were employed as targets, no additional effector cells were recruited. These results indicate that the enhanced cytotoxicity induced by MoAb B1B6 is the result of either recruitment of new effector lymphocytes or of an increased recycling capacity of preexisting effector cells. Together with previous observations, these findings support the conclusion that LSGP belongs to the set of surface molecules which regulate human lymphocyte activation. PMID- 2969142 TI - Characterization and expression of the human T cell receptor-T3 complex by monoclonal antibody F101.01. AB - A murine monoclonal antibody (MoAb) F101.01 reacting with the T cell receptor (TCR)-T3 complex is presented. Immunohistological studies showed that F101.01 specifically stains T-zone lymphocytes in lymph nodes, tonsils, and splenic tissue. Two-colour immunofluorescence and flow cytometry demonstrated co expression of the antigen defined by F101.01 and the pan-T cell antigens defined by CD2, CD3, CD5, and CD7 antibodies. Cells stained with CD4 and CD8 antibodies were both included in the F101.01-positive population, whereas CD16-positive natural killer cells (NK), B cells (CD19 and CD20), and myeloid cells (CD13 and CD33) were excluded. The target antigen of F101.01 co-modulated with the CD3 defined antigen (T3) and the TCR recognized by the MoAb WT-31. CD3 antibody and WT-31 both blocked binding of F101.01. F101.01 precipitated the TCR-T3 complex from lysates of 125I-labelled peripheral blood mononuclear cells (PBMC) and HPB ALL, when the lysate was prepared with a detergent (digitonin) that conserves the TCR-T3 complex. FACS analysis of T cells from a patient with a T cell immunodeficiency demonstrated that delta-TCS-1-CD3+CD4+ and delta-TCS-1-CD3+CD8+ cells were brightly F101.01+, whereas a large subpopulation of delta-TCS 1+CD3+CD4-CD8- cells were weakly F101.01+. We conclude that F101.01 recognizes a conformational epitope of the TCR-T3 complex and that it reacts with the alpha beta TCR-T3 and the gamma delta TCR-T3 complexes with different intensities. PMID- 2969143 TI - Prevention of occupational back disorders--an intervention study. PMID- 2969144 TI - Atlas of standardized low-back tests and measures of the National Institute for Occupational Safety and Health. PMID- 2969145 TI - [Biochemistry and pathophysiology of the erythrocyte membrane skeleton]. PMID- 2969146 TI - [Recent advances in the molecular biology of cardionatrin]. PMID- 2969147 TI - Model of Huntington's disease. PMID- 2969148 TI - Ways of removing essential differences between polyclinical and ambulatory assistance level for town population and countryside population. PMID- 2969149 TI - [An inhuman society without handicapped. Interview by Lars Peter Bergqvist]. PMID- 2969150 TI - [Circulating immune complexes and cellular cytotoxic reactions in Sjogren's syndrome]. PMID- 2969151 TI - Use of [3H]trimetoquinol as a radioligand in human platelets: interaction with putative endoperoxide/thromboxane A2 receptor sites. AB - We have recently shown that trimetoquinol [1-(3,4,5-trimethoxybenzyl)-6,7 dihydroxy-1,2,3,4- tetrahydroisoquinoline; TMQ] is a potent and stereoselective [R(+)-isomer greater than S(-)-isomer] antagonist of aggregation induced by thromboxane A2 and stable epoxymethano PGH2 analogues (U46619; U44069) in human platelets. The present study was undertaken to characterize the pharmacological specificity of binding sites for racemic- [3H]TMQ in washed human platelets. Specific binding of [3H]TMQ, determined by addition of 100 microM R(+)-TMQ, accounted for 12-26% of the total binding at 1.0 microM. Saturation data suggested two binding sites with apparent dissociation constants of 2.8 nM and 1.4 microM for the high and low affinity binding sites, respectively. Maximal binding densities (pmol/mg protein) were 2.3 and 42.9 for the respective high and low affinity sites. The optical isomers of TMQ, cis and trans-isomers of 13 azaprostanoic acid (13-APA), and U46619 inhibited [3H]TMQ specific binding to the low affinity site. R(+)-TMQ was 32-fold more potent than S(-)-TMQ at inhibiting [3H]TMQ binding, and the stereoselective potency difference and concentrations needed to inhibit binding were similar to those required for inhibition of U46619 induced platelet aggregation and serotonin secretion. Similarly, trans-13-APA was 90-fold more potent than cis-13-APA as a competitor of [3H]TMQ binding. U46619 also inhibited TMQ binding at concentrations (IC50 = 0.4 microM) similar to those required for aggregation (EC50 = 0.16 microM) and secretion (EC50 = 0.23 microM). At 1 mM, arachidonic acid inhibited [3H] TMQ binding by only 25%. Our studies indicate that [3H]TMQ interacts with specific binding sites with characteristics indicative of putative endoperoxide/thromboxane A2 receptors in human platelets. PMID- 2969152 TI - Specific suppression of allograft rejection after treatment of recipient mice with ultraviolet radiation and allogeneic spleen cells. AB - Sensitization of UV-irradiated mice with alloantigen results in suppression of the immune response against that antigen. The induction of delayed hypersensitivity and the proliferative response of spleen cells isolated from the UV-irradiated mice are significantly suppressed. We demonstrate here that the rejection of tissue allografts is also suppressed after treatment of the recipient animals with UVR. Two signals, UVR and antigenic sensitization are required to suppress allograft rejection, exposure to UVR alone is not sufficient. The resulting immunosuppression is specific for the antigen used to sensitize the UV-irradiated animal. The induction of graft-versus-host disease was also significantly suppressed when spleen cells from UV-irradiated alloantigen sensitized mice were used to reconstitute lethally irradiated allogeneic mice. Our interpretation of these data is that the antigen-specific suppressor T cells present in the spleens of the UV-irradiated alloantigen sensitized mice are suppressing the immune response against the alloantigen. Thus, the induction of suppression by UVR serves as a novel method of suppressing graft rejection. PMID- 2969153 TI - Stimulation of cell replication in transplanted pancreatic islets by nicotinamide treatment. AB - A major obstacle for islet transplantation in insulin-dependent diabetes mellitus is to obtain a sufficient amount of islet tissue. This may partly be overcome if the cell replication in the grafted islet preparation could be stimulated in the recipient. In the present study adult mice were treated for 14 days with daily injections of nicotinamide (500 mg/kg body weight) or saline. Subsequently, the autoradiographic labeling index in the cells of the pancreatic islets were calculated in normal mice and in syngeneic islets transplanted into alloxan diabetic mice. Treatment with nicotinamide caused a more than three-fold increase in the islet cell labeling index in the endogenous pancreatic islets, and also a 50% increase in the cell replication rates of the transplanted islets. It thus appears that nicotinamide enhances islet cell replication, possibly through an inhibition of poly(ADP-ribose) synthetase activities. PMID- 2969154 TI - Donor class I and class II major histocompatibility complex antigen expression following liver allografting in rejecting and nonrejecting rat strain combinations. AB - Orthotopic liver allografts in the nonrejecting DA-to-PVG strain combination and in the DA-to-LEW strain combination were studied at various times after transplantation for donor class I and class II MHC expression using immunohistological techniques and quantitative analyses. DA-to-DA isografts were also studied. In the isografts, weak class I induction on hepatocytes and biliary epithelium was noted from day 5, and this persisted to day 15, the last time point examined. In DA-to-PVG allografts, class I induction also appeared on hepatocytes and biliary epithelium from day 5, but was more intense than in the isografts. Nevertheless, the induction was patchy within most grafts, and in some grafts was not prominent. Quantitative absorption analyses demonstrated that the maximum increase in donor class I expression was only 3-fold over the normal liver. In the strong DA-to-LEW combination, class I induction on hepatocytes seemed to appear earlier, beginning at day 3, and was more uniform and intense than in the DA-to-PVG model from day 5. In the isografts, there was no induction of class II antigens on hepatocytes or biliary epithelium at any stage, but from days 5 to 15 there was a marked increase in the number of isolated, class II positive cells in the hepatic lobule, probably representing class II induction in the Kupffer cells of the isografts. In DA-to-PVG allografts, biliary epithelium became class II-positive from day 5, and this persisted to day 30, the last time point examined. Weak but definite class II induction was seen on some hepatocytes from day 5 through day 30. However, the majority of hepatocytes remained class II negative. By day 30, there was virtually no donor class II staining the sinusoids, but isolated class II-positive cells of recipient type were seen, the pattern suggesting a replacement of the graft Kupffer cells by recipient Kupffer cells at this stage. By quantitative absorption analysis, donor class II expression in the grafts increased approximately 5-fold. In DA-to-LEW allografts, class II induction was not noticeably different from that seen in the DA-to-PVG model, except that induction of class II antigens on the Kupffer cells possibly appeared earlier in this strain combination. PMID- 2969155 TI - [Interrelations of NAD and adenosine transformation in the rat liver]. AB - The interrelationship of NAD and adenosine (inosine) conversions in the rat liver is investigated. The ratio of products of NAD+ conversions (ADP-ribose, inosine, hypoxanthine and ribose phosphates) are established. AMP and adenosine are not detected, which indicates an availability of different activities of the corresponding enzymes. It is shown that under conditions of the high inorganic phosphate concentration (33 mM) ribose-1-phosphate, formed in the purine nucleoside phosphorylase reaction, is accumulated due to the phosphoribomutase inhibition, but in the presence of NAD+ the utilization of ribose phosphate increases significantly. Nicotinamide inhibits the NAD+-glycohydrolase reaction in the system containing 33 mM phosphate, NAD+ and adenosine and simultaneously it lowers the utilization of ribose. PMID- 2969156 TI - Controlling lungworm. PMID- 2969157 TI - The Arthus reaction in domestic cats. AB - A classic Arthus reaction was elicited in normal domestic cats using chicken red blood cells as antigen. The response was quantitated grossly by measuring the area of the resulting skin bleb at several set time intervals and by microscopic examination of biopsies taken at the conclusion of each of the trials. This method produced an intense Arthus reaction in each of the cats tested. PMID- 2969158 TI - [The reproductive function and the risk of developing cancer of the breast in different ethnic groups]. AB - The study was concerned with the relationship between certain characteristics of reproductive case history and risk for breast cancer development in three different ethnic groups of females living in the Moldavian SSR: Moldavians, Ukrainians and Russians. Risk of cancer was found to be higher in all groups in cases of premenstrual syndrome, late onset of sexual life (greater than or equal to 26 years), suspension of sexual life for more than 12 months, late first pregnancy and delivery, low productivity (0-1 birth in case history), agalactia, and menstrual cycle resumption 1-2 months after childbirth. However, certain ethnic differences in relationships between such factors as age of menopause onset, no pregnancies, late pregnancy (greater than or equal to 40 years) and type of contraceptives used and risk of cancer are rather due to the difference in prevalence of said factors in the groups under study. The results of the investigation suggest a common pattern of influence of reproductive case history factors on risk for breast cancer in different ethnic groups. PMID- 2969159 TI - [Spontaneous tumors in rats bred at the N.N. Petrov Research Institute of Oncology of the USSR Ministry of Health]. AB - The paper discusses spontaneous tumor profile in albino rats bred at the N. N. Petrov Research Institute of Oncology of the USSR Ministry of Health (1981-1984). The overall incidence of tumors and that of malignant ones were shown to be 27.2 and 12.5%, respectively, in males and 47.4 and 16.4%, respectively, in females. Pituitary and hemopoietic tumors most frequently occurred in males (67%). Female rats most often revealed tumors of the pituitary, breast and hemopoietic system (90%). On the whole, the tumor profile proved identical in the animal population studied and that of Rappolovo farm of the USSR Academy of Medical Sciences. However, infection resistance in the former population was superior. PMID- 2969160 TI - [A complex of automated systems for processing information in an oncologic service]. AB - The increasing amount of medical and executive work of oncologic institutions stimulates the development of automated information systems for evaluation of cancer service performance, its management and planning. A complex of automated information systems was developed and integrated with practice. It is based on the automated system for processing information of a republican cancer service patterned after a population cancer registry. It also incorporates systems for controlling cancer patients follow-up, information retrieval, cancer statistics prognosis, modelling of planning of development and control of cancer service effectiveness and an automated bank of data. PMID- 2969161 TI - [Mechanism of the action of plasmin in myocardial damage due to coagulation]. PMID- 2969162 TI - [Treatment with cardiac glycosides and metabolically active preparations of patients with chronic cor pulmonale]. PMID- 2969163 TI - [Circulatory failure in patients with chronic cor pulmonale]. PMID- 2969164 TI - [Diagnosis and surgical therapy of suppurative inflammations of the adnexa]. AB - A report is given on 119 patients suffering from abscessforming adnexitis, mainly tubo-ovarian abscesses (40.3%) and pyosactosalpinges (38.7%). These women were operated on within the first 24 hours after admission to the hospital. Before the operation was performed 70.6% of the women had been submitted to laparoscopy. The other ones (28.4%) had unequivocal primary sign of the disease. Laparoscopic diagnosis is recommended because especially in the early phase of abscess formation no clear diagnosis by clinical examination and paraclinical findings is possible. Operation as early as possible in case of formation of a tubo-ovarian abscess is recommended by reason of the good experience and results which become apparent by small intra- and post-operative complication rate of 1.7% respectively, the short hospital stay of 12.8 days and the short time of risamblement of 53 days. The extent of the operation should depend on degree of severity of the illness, reproductive aspects. PMID- 2969165 TI - [Incidence of postoperative adhesions following use of various methods of coagulation. An animal experiment study]. AB - In 5 groups of 20 rats each, equal areas of the uterus horns and of the corresponding peritoneum of the anterior abdominal wall were coagulated resp. vaporized. To achieve this endocoagulation, mono- and bipolar high frequency current, and CO2- and Neodym-YAG-lasers were used. After 2 resp. 4 weeks, no intraabdominal adhesions were found in two animals after endocoagulation and in one animal each after coagulation with bipolar high frequency current and with CO2-laser. The lowest adhesion rates were seen after endocoagulation and coagulation with bipolar high frequency current. After application of the YAG laser, to the uterus horns adhesions were present in 100%, applying the YAG-laser to the anterior abdominal wall for a short time there were produced few adhesions. After coagulation resp. vaporization of equal areas of the anterior abdominal wall, significantly more adhesions were found after CO2-laser application, compared to all other techniques (p less than 0.01). Adhesions to the abdominal wall were significantly less frequent than adhesions to the tubes (p less than 0.01) with all techniques except with the CO2-laser. PMID- 2969166 TI - Therapeutic activity of teicoplanin on experimental listeriosis compared with that of vancomycin and ampicillin. AB - Several strains of Listeria monocytogenes and other Listeria spp. are without exception susceptible to teicoplanin (MIC 0.25 mg/l). Vancomycin as well as ampicillin are likewise active. A bactericidal effect of teicoplanin was only achieved at rather high concentrations and after incubation of several hours. There is no synergistic effect between teicoplanin and gentamicin. The therapeutic activity of teicoplanin as well as vancomycin in mice infected with L. monocytogenes is low. The efficacy of ampicillin could not be achieved. Treatment of chronically infected athymic, nude mice with teicoplanin is ineffective. Consequently, teicoplanin is not able to replace ampicillin in the therapy of listeriosis. PMID- 2969167 TI - [HLA-B27 antigen in the nonspecific back pain type of spondylopathy]. PMID- 2969169 TI - Should oxygen be administered after laparoscopy in healthy patients? AB - This study aimed to assess the oxygen flow necessary to maintain satisfactory oxygen saturation when administered via a nasopharyngeal catheter. Oxygen saturation was displayed by a pulse oximeter and/or measured in arterial blood samples. Thirty-six healthy women scheduled for elective diagnostic laparoscopy were anaesthetized using thiopentone, fentanyl and O2/N2O. Atracurium was used as relaxant which was reversed with atropine and neostigmine. Arterial samples were obtained prior to anaesthesia, on arrival in the postoperative ward and 1 h postoperatively. Oxygen saturation was monitored postoperatively using a pulse oximeter. The patients were randomly divided into three groups which received either no oxygen, 2 l O2/min or 4 l O2/min. On arrival in the postoperative ward 15% of the patients were below the normal limit of O2 saturation (94%). In patients receiving 2 l or 4 l O2, oxygen saturation was well above normal values. In patients receiving no oxygen, two had low oxygen saturation (92% and 93%). Comparing saturation values obtained in arterial samples with values measured with pulse oximetry gave r = 0.79. It is concluded that all patients should be given oxygen in the immediate postoperative period. Increasing oxygen flow from 2 to 4 l/min had no major effect on oxygen saturation. These results were obtained in healthy patients following minor abdominal surgery. PMID- 2969168 TI - Desogestrel in monophasic E/P combination: monitoring of endocrine parameters and evaluation of hemostatic function. AB - The Authors evaluated modifications in androgenic pattern as well as in parameters of androgenic function and coagulation in a group of women treated with a combination contraceptive containing Ethinyl estradiol and desogestrel. PMID- 2969170 TI - Histological changes of bile duct in experimental graft-versus-disease across minor histocompatibility barriers. III. Immunoelectron microscopic observations. AB - Graft-versus-host disease (GVHD) was produced by injecting lymphoid cells from B10.D2 mice into BALB/C mice. Both are H-2-identical but differ only at multiple minor H loci. The expression and localization of MHC class II antigens on the bile duct epithelium was examined using an immunoelectron microscopical method. All GVHD mice developed bile duct lesions of chronic nonsuppurative cholangitis and expressed MHC class II antigens on their bile duct epithelium, while none of the control mice, which injected with the same number of syngeneic lymphocytes, developed bile duct lesions or expressed the antigens. The antigenic expression was characteristically localized on the basolateral surface of the bile duct epithelium but not on the apical surface. Furthermore, the expression varied markedly in its intensity and distribution within the same liver and even within a single bile duct. The infiltration of Lyt-1-positive helper/inducer lymphocytes in the duct epithelial layer apparent by electron microscopy was predominant to a much degree than that of Lyt-2-positive cytotoxic/suppressor lymphocytes and non lymphocytic cells. The immunological mechanisms involving helper/inducer T cells in association with MHC class II antigens on bile duct epithelium may be important in the induction and progression of the bile duct lesions apparent in the present GVHD model. PMID- 2969171 TI - An autopsy case of idiopathic enlargement of the right atrium, and a review of the literature. AB - A 69-year-old man in whom idiopathic enlargement of the right atrium was revealed at autopsy is described. The patient had had cardiomegaly of at least 19 years' duration prior to his death, even though cardiac symptoms were absent. Cause of death was pancreatic carcinoma. Postmortem examination revealed marked and diffuse dilatation of the right atrium and moderate dilatation of the left atrium. Measurement of the cardiac chambers showed that the right and left atria were 7.6 and 4.7 times as large as those of normal hearts, respectively. The volume of either ventricle was about twice the normal value. Histologically, widespread cardiac muscular degeneration and necrosis, diffuse fibrosis, and focal lymphocytic infiltration were found in the right atrium and also, to a lesser degree, in the left atrium. Such pathologic changes were not found in either of the ventricles. The etiology of these muscular changes, which might have been related to atrial enlargement, was unclear. The present case was thought to be consistent with idiopathic enlargement of the right atrium, and a brief review of the literature is given. PMID- 2969172 TI - Open trial of mesoglycan in the treatment of cerebrovascular ischemic disease. PMID- 2969173 TI - T-cell clones specific for synthetic polypeptides in cellular collaborations. PMID- 2969174 TI - Modulation of T-cell functions by peptide hormones. PMID- 2969175 TI - Intracellular traffic of the mannose 6-phosphate receptor and its ligands. AB - The role of the mannose 6-phosphate recognition marker and the phosphomannosyl receptor in the intracellular transport of acid hydrolases is well established. Several details of the process, however, are presently unclear and warrant further investigation (Table 1). The development of in vitro systems for the reconstitution of receptor-mediated endocytosis and intracellular transport, together with the application of the techniques of molecular biology to this field, should lead to major advances in our understanding of the targeting of enzymes to lysosomes. PMID- 2969176 TI - Correlations between histamine and opioid peptides on the modulation of inflammatory processes in rats exposed to stress. AB - The role of mast cell histamine in body reactivity of rats under experimental stressful conditions was studied. Animals submitted to chronic anaphylactoid reactions (by injecting compound 48/80 at the dose of 1 mg/kg, i.p., twice daily, for five days), when exposed to cold-restraint stress, exhibited a fully evident inflammatory response in the carrageenin-oedema test, whereas saline-treated rats, under the same experimental conditions, showed reduced paw oedema. Interestingly, a single injection of compound 48/80 increased the pituitary content of Beta-endorphin(ir), but chronic administration failed to produce this effect suggesting that some adaptation of the organism to repeated anaphylactoid reactions may occur. These results support the hypothesis of correlations between pituitary Beta-endorphin and mast cell histamine in the reactivity of the organism to stressful stimuli. PMID- 2969177 TI - Percutaneous transhepatic recanalization and thrombolysis of the superior mesenteric vein. PMID- 2969178 TI - Subclavian artery steal syndrome: treatment by percutaneous transluminal angioplasty. AB - We present our experience with 24 patients in whom percutaneous transluminal angioplasty was performed in the proximal subclavian artery. Seventeen patients had symptoms of subclavian steal syndrome, and seven had symptoms of upper extremity ischemia. Eighteen (75%) had excellent clinical results, with 80-100% restoration of the expected lumen diameter. Three patients required repeat dilatation because of recurrent symptoms. The only complication was a single case of brachial artery occlusion. Our results suggest that subclavian artery angioplasty is a safe and effective method for treating subclavian artery steal syndrome and upper extremity ischemia. PMID- 2969179 TI - Regulator soluble factors of human IgE synthesis: II. Biological activity. AB - In view of the experiments carried out on rats by various investigators in order to identify and characterize soluble factors that regulate IgE synthesis, we decided to apply these systems to humans through different "in vitro" techniques, and in different population groups. After its production through bidirectional mixed cultures as well as its isolation by column of Sepharose Con-A (affinity chromatography), we proceeded to confirm its biological activity. The modifications produced by these factors on the synthesis of IgE "in vitro" (modification of Phadebast IgE PRIST radioimmunoanalysis technique for low levels) on lymphocytes of controls, atopics and pollinosis with and without immunotherapy were confirmed, as well as the non-modification on IgG synthesis. A considerable increase (greater than 300%) in IgE production in lymphocytes of healthy subjects was observed in relation with the stimulating factor obtained in CML of atopic lymphocytes. PMID- 2969180 TI - The stimulatory effect of a single intravenous dose of propranolol of some immune parameters in humans. AB - Propranolol (PRO) is a beta-adrenergic receptor antagonist. The studies were aimed to evaluate the effect of a single intravenous PRO injection at a dose of 0.1 mg/kg on some immune parameters, and were performed on 12 men, including 9 duodenal ulcer patients and 3 healthy subjects, aged 19-23 years (mean 20.5 +/- 1.2). Blood samples were drawn before, 30 and 150 min following the injection. Results obtained in duodenal ulcer patients and healthy subjects revealed the same type of changes. A single PRO injection did not significantly affect total peripheral blood lymphocyte, Th cell and monocyte counts; however, it slightly but significantly lowered numbers of T and Ts cells, which was accompanied by a rise in Th/Ts cell number ratio and NK cell numbers. PRO significantly augmented spontaneous as well as PHA-stimulated IL-2 receptor expression, IL-2 generation, NK cell activity and AMLR observed at 30 and 150 min after the drug injection. Possible mechanisms of these changes are discussed. PMID- 2969181 TI - Workers' compensation. PMID- 2969182 TI - The epidemiology of resting heart rate in a national sample of men and women: associations with hypertension, coronary heart disease, blood pressure, and other cardiovascular risk factors. AB - Few reports have been published on the epidemiology of heart rate in general populations including women and blacks. In the first cycle of the National Health Examination Survey, resting heart rate was significantly higher in women than in men at each age in both races. Age and race were inconsistently related to heart rate, black men having slightly lower rates than white men at ages 18 to 34. The product of heart rate and mean blood pressure was consistently highest in black women. Heart rate was associated with obesity, waist girth, and waist-to-hip girth ratio in men under 55 years and with blood pressure and post-load serum glucose in all groups. Sex differences at each age were larger (6 to 14 beats/min) when adjusted for multiple variables. Multivariate linear regression analysis confirmed the independent associations of heart rate with blood pressure and serum glucose. After controlling for other risk factors, prevalent hypertensive subjects had higher heart rates than normotensive individuals in all groups. Further research is needed to elucidate mechanisms of the associations of resting heart rate with sex, risk factors, coronary heart disease, and mortality. PMID- 2969183 TI - Sudden increase in left ventricular mass secondary to acute myocarditis. PMID- 2969184 TI - Basal and exercise plasma levels of atrial natriuretic factor in congestive heart failure. PMID- 2969185 TI - No-reflow phenomenon during percutaneous transluminal coronary angioplasty. PMID- 2969186 TI - Effects of antihypertensive therapy on cardiac function. AB - The failure of clinical trials in the treatment of hypertension to show protection against mortality from coronary heart disease has prompted a major reconsideration of the indications and modes of therapy. One area of concern is the lipid disturbances seen with diuretics and non-ISA (intrinsic sympathomimetic activity) beta blockers; another is the negative inotropic effect of beta blockers. The more widespread use of vasodilators--either alpha-blockers, angiotensin-converting enzyme (ACE) inhibitors, or calcium channel blockers- therefore seems appropriate. Among the calcium channel blockers, nicardipine appears to be a particularly attractive choice, in part because of its lack of negative inotropism. PMID- 2969187 TI - Left ventricular hypertrophy: its prime importance as a controllable risk factor. AB - Left ventricular hypertrophy, which is defined as an abnormal increase in the mass of the left ventricle, has become recognized as an important independent risk factor for predicting several untoward cardiac events, including myocardial infarction, congestive heart failure, and sudden death. The presence of left ventricular hypertrophy is difficult to detect by electrocardiography but can be accurately assessed by echocardiography. Left ventricular hypertrophy can be detected in about 5% of all hypertensive patients by electrocardiography but is found in about 50% by echocardiography. Although it is possible to restore left ventricular mass to normal by controlling hypertension with pharmacotherapy, several commonly prescribed antihypertensive medications, including diuretics and vasodilators, have failed to demonstrate the ability to reverse left ventricular hypertrophy. Other agents, including most beta-blockers, have been found to reverse it. Reversal may take 18 to 24 months from the initiation of therapy. This reversal may be accompanied by an increase in cardiac index, reductions in total peripheral resistance, and improvements in the diastolic properties of the left ventricle. It has yet to be determined whether these improvements reduce the risk of myocardial infarction, heart failure, and sudden death. However, it is likely that some antihypertensive medications may be superior to others in reducing long-term cardiac risks. PMID- 2969188 TI - Cardiac function in experimental hypertension. PMID- 2969189 TI - Regional and temporal variation in oscillatory blood flow in sickle cell disease. AB - We examined 55 patients with sickle cell anemia (HbSS) and 16 with hemoglobin SC disease (HbSC) to ascertain the presence of periodic microcirculatory flow (PMF) using laser-Doppler velocimetry. Forty-nine percent of patients with HbSS and 12.5% of HbSC patients had PMF on at least one occasion and in at least one site. The presence of PMF could be correlated with an increase in packed cell volume (PCV) and hemoglobin in individuals with HbSS and an increased white blood cell and platelet count in patients with HbSC. PMF may result from complex interactions among perfusion pressure, capillary tone, and the numbers and intrinsic properties of formed blood elements. PMID- 2969190 TI - Sonographic demonstration of hypoplasia of the middle phalanx of the fifth digit: a finding associated with Down syndrome. AB - Sixty percent of neonates with Down syndrome have hypoplasia of the middle phalanx of the fifth digit. We demonstrate this finding sonographically between 17 and 20 weeks' gestation in four of five fetuses with Down syndrome. The appearance of this abnormal phalanx as well as the abnormal curvature of the fifth digit is described. PMID- 2969191 TI - Plasma levels of atrial natriuretic peptide during normal pregnancy and in pregnancy complicated by hypertension. AB - To clarify a possible role for atrial natriuretic peptide in the pathophysiology of pregnancy complicated by hypertension, we studied plasma levels of atrial natriuretic peptide in 176 pregnant women with or without hypertension. Plasma atrial natriuretic peptide levels in normal pregnant women showed a gradual increase as pregnancy advanced, but the mean (+/- SD) concentrations in women in each trimester (34.8 +/- 14.7 pg/ml in the first trimester, n = 35; 38.7 +/- 12.2 pg/ml in the second trimester, n = 34; and 43.1 +/- 20.0 pg/ml in the third trimester, n = 71) did not differ statistically from the mean plasma atrial natriuretic peptide level in nonpregnant women (38.2 +/- 13.6 pg/ml, n = 44). In contrast, plasma atrial natriuretic peptide levels were elevated in 9 of the 12 women who had hypertension. The mean plasma atrial natriuretic peptide concentration in these patients (162 +/- 95.2 pg/ml) was significantly (p less than 0.01) higher than in normal pregnant women and in nonpregnant controls. On the other hand, 11 pregnant women with proteinuria or edema but without hypertension had normal plasma atrial natriuretic peptide levels. These results suggest that plasma atrial natriuretic peptide levels are normal in women during uncomplicated pregnancy, while the levels are elevated in pregnancy complicated by hypertension. Increased atrial natriuretic peptide secretion in the latter condition may reflect a mechanism of compensation that operates in response to water and sodium retention. PMID- 2969192 TI - Beta-endorphins in ovarian follicular fluid. PMID- 2969193 TI - Attitudes of occupational therapy personnel toward persons with disabilities. AB - The study described in the following article investigated the attitudes of occupational therapy personnel toward persons with disabilities. The examination involved attitudes in general as measured by the Scale of Attitudes Toward Disabled Persons (SADP) (Antonak, 1981), specific attitudes toward infants with Down's syndrome, beliefs concerning the importance of favorable attitudes toward patients, and beliefs concerning the role of the American Occupational Therapy Association (AOTA) in the development of ethical guidelines for the delivery of health care services. The results of a survey of 619 occupational therapy personnel indicate that they hold a very favorable attitude toward persons with disabilities, and that they believe that a negative attitude would adversely affect the therapeutic relationship. The majority agree that the expression of a favorable attitude should be a criterion in student selection. The majority also agree that AOTA should develop position papers on ethical issues and take a public position on issues pertinent to the rights of the disabled person. Occupational therapy personnel proved to be very homogeneous on the general attitudinal scale. However, the Down's syndrome scenario uncovered significant differences among the different categories of respondents in the variables for professional level, area of practice, years of practice, and geographic location. A majority of respondents agreed that is is unethical to withhold needed surgery from a child because of disability. However, they differed on the ethics of aborting a fetus with Down's syndrome (most thought it was unethical). PMID- 2969194 TI - Additive and synergistic interaction of atrial natriuretic peptide and volume expansion. AB - To investigate whether atrial natriuretic peptides (ANP) are entirely responsible for the natriuresis of volume expansion (VE), we studied the natriuresis from acutely snared and nonsnared kidneys of rats undergoing sustained saline VE and subsequent infusion of maximal doses of ANP (atriopeptin II), or vice versa. Fractional excretion of Na (FENa) was increased from control (0.15 +/- 0.03%) to 4.6 +/- 0.6% by VE and to 10.9 +/- 1.5% by subsequent ANP infusion. With ANP alone FENa increased from control (0.24 +/- 0.04%) to 2.6 +/- 0.3%, and to 10.8 +/- 0.9% with VE, showing additive effects by both. Natriuresis with VE was significantly greater than with ANP alone (P less than 0.01) and both exhibited synergism, producing significantly greater natriuresis in the presence of the other than alone (P less than 0.05 for both). A reduction in perfusion pressure inhibited the effects of ANP and reduced that of VE. Natriuresis was produced by ANP independently of changes in glomerular filtration rate. It is concluded that 1) VE and ANP produce natriuresis by different renal mechanisms and that therefore, ANP can only account for part of the natriuresis of sustained VE; 2) the two have a synergistic interaction on natriuresis, probably through an intrarenal mechanism. PMID- 2969195 TI - Cardiovascular responses to microinjection of ANF into dorsal medulla of rats. AB - Atrial natriuretic factor (ANF) has been suggested as a putative neurotransmitter in central pathways involved in the control of the cardiovascular system. To investigate this possibility, 50 nl of 10(-7) M ANF were microinjected into discrete sites in the nucleus of the tractus solitarius (NTS) where baro- and chemoreceptor afferents terminate. Injections into 36 of a total of 66 sites in the NTS of paralyzed artificially ventilated Wistar rats under urethan anesthesia were found to produce a significant decline in heart rate [HR; -9.2 +/- 2.9 (SE) beats/min, P less than 0.05] and mean arterial pressure [MAP; -11.1 +/- 1.2 (SE) mmHg, P less than 0.01]. Similar responses were also present in anesthetized animals breathing spontaneously. Microinjection of an inactive peptide analogue or of saline did not produce cardiovascular changes. It was also found that ANF injection into the cuneate nucleus (20 of 38 sites) and the spinal trigeminal complex (28 of 42 sites) produced a decrease in MAP and HR that were of the same magnitude as those seen in the NTS. Injections of ANF into the medial longitudinal fasciculus (n = 22), hypoglossal nucleus (n = 9), area postrema (n = 16), and dorsal motor nucleus of the vagus (n = 11) did not change HR or MAP. These results suggest that ANF may serve as a neurotransmitter involved in cardiovascular reflexes mediated by specific nuclei in the dorsal medulla. PMID- 2969196 TI - Vagal C-fiber blockade abolishes sympathetic inhibition by atrial natriuretic factor. AB - Administration of atrial natriuretic factor (ANF) to anesthetized rats decreases renal nerve activity (RNA), an effect prevented by vagotomy but not by atropine. We sought to determine whether afferent vagal C-fibers mediate the inhibition of sympathetic outflow. ANF (2.5 micrograms/kg iv) decreased mean arterial pressure (MAP) by 25 +/- 2 mmHg, RNA by 11 +/- 5%, and least splanchnic nerve activity (LSNA) by 10 +/- 4% in anesthetized rats with arterial baroreceptors intact, and by 40 +/- 3 mmHg, 28 +/- 7%, and 23 +/- 4%, respectively, in sinoaortic denervated rats. Inhibition of RNA and LSNA by ANF was reduced slightly by cooling the vagi to 6 or 7 degrees C, a temperature at which conduction in A fibers was blocked and that in C-fibers attenuated; inhibition was abolished when C-fibers were blocked by cooling to 0 degrees C. We conclude that the inhibition of RNA and LSNA by ANF was mediated by afferent vagal C-fibers. We also obtained evidence that the aortic nerves contribute to ANF-induced inhibition of RNA. Our results support the notion that ANF evokes a generalized decrease in sympathetic tone that contributes to the hypotension, cardiac inhibition, and natriuresis accompanying systemic administration of the peptide. PMID- 2969197 TI - Cardiorenal endocrine dynamics during volume expansion in hypothyroid dogs. AB - To address the role of atrial natriuretic factor (ANF) in hypothyroidism in the control of cardiorenal-endocrine function during volume loading, the relationships between atrial pressure, ANF, the renin-angiotensin-aldosterone system, and renal hemodynamic and excretory function were examined during and after acute 10% body wt saline volume infusion in pentobarbital-anesthetized hypothyroid dogs (n = 8). Hormonal changes before and after thyroidectomy were also evaluated. Four to 6 wk after thyroidectomy, ANF decreased and arginine vasopressin (AVP) and plasma renin activity (PRA) increased. Acute saline volume expansion caused an increase in ANF and decreases in AVP and PRA. Atrial pressure increased throughout volume expansion. Despite the absence of an increase in glomerular filtration rate (GFR) during volume expansion, urinary sodium excretion increased due to a marked rise in fractional excretion of sodium. These studies demonstrate that in hypothyroidism 1) ANF is decreased; 2) despite the decrease in basal ANF, increases in atrial pressure can stimulate relase of ANF; 3) despite the absence of an increase in GFR during volume expansion, fractional excretion of sodium increases associated with an increase in ANF; and 4) a lack of an increase in GFR during volume expansion is not related to an inability to increase ANF. PMID- 2969198 TI - Female hypersexuality treated with cyproterone acetate. PMID- 2969199 TI - Psychiatric disorder in a sample of the general population with and without chronic medical conditions. AB - The authors studied data on psychiatric disorders and eight chronic medical conditions in a community sample of 2,554 persons. The sex- and age-adjusted prevalence of any psychiatric disorder in the preceding 6 months was 24.7% and of lifetime psychiatric disorder was 42.2% among persons with one or more medical conditions, compared to 17.5% and 33.0%, respectively, for persons with no medical condition. Persons with chronic medical conditions were more likely to have lifetime substance use disorders and recent affective and anxiety disorders. Arthritis, cancer, lung disease, neurological disorder, heart disease, and physical handicap were strongly associated with psychiatric disorders, but hypertension and diabetes were not. PMID- 2969200 TI - An affinity gel for the inhibition, binding, and isolation of serine proteases. AB - A preparation procedure for gels for the specific binding and inhibition of serine proteases is described. Phosphoryl trifluoride was synthesized and reacted with two different types of agarose gels, a crosslinked agarose (Sepharose CL-4B) and an agarose containing spacer arms with terminal vicinal-diol groups (a hydrolyzed epoxy-activated Sepharose 6B). The phosphoryl difluoride groups coupled to the gels were, in both cases, further modified by treatment with isopropanol to obtain isopropyl fluorophosphate groups covalently bound to the matrix. It was found that both modified gels absorbed and inhibited plasmin, but that the modified gel with spacer arms was markedly more efficient. PMID- 2969201 TI - The quantitative discrimination of corneal type I, but not skeletal type II, keratan sulfate in glycosaminoglycan mixtures by using a combination of dimethylmethylene blue and endo-beta-D-galactosidase digestion. AB - The quantitation of individual glycosaminoglycans in mixtures of polyanions using the dimethylmethylene blue (DMB) method described by R. W. Farndale, D. J. Buttle, and A. J. Barrett (1986, Biochim. Biophys. Acta 883, 173) is dependent on enzymatic hydrolysis by specific polysaccharidases. While using this method to examine the keratan sulfate (KS) of the intervertebral disc we found that digestion with commercially available keratanase decreased binding to DMB by less than 30%, whereas corneal KS was reduced by 85%. However, by preincubating the KS fractions with endo-beta-D-galactosidase prior to keratanase treatment the corneal KS could be completely digested and disc KS digestion increased to 60%. It is suggested that the resistance of the disc KS to these digestive procedures arises from branching and/or sites of multisulfation on the polysaccharide chain. Agarose gel electrophoresis and compositional analyses of the keratan sulfates supported such an interpretation. PMID- 2969202 TI - A method for the determination of N-acetylglucosaminyltransferase III activity in rat tissues involving HPLC. AB - A fluorescence assay method for UDP-GlcNAc:glycopeptide beta 1-4 N acetylglucosaminyltransferase (Gn T-III) has been developed involving a pyridylaminated sugar as a substrate. A fluorescent sugar chain, in which the reducing end of the GlcNAc beta 1-2Man alpha 1-6(GlcNAc beta 1-2Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4GlcNAc has been aminated with 2-aminopyridine, is incubated with an enzyme sample, and then the fluorescent product with a bisecting N-acetylglucosamine is separated by reverse-phase high performance liquid chromatography and quantitated with a fluorescence detector. This assay method was found to be sensitive enough for the detection of 0.1 pmol of a reaction product. This assay is a reliable alternative to the use of a radiolabeled substrate and can be used for assaying N acetylglucosaminyltransferase activity in crude extracts of various rat tissues. The kinetic experiments were carried out using crude enzyme extracts from the rat kidney. The enzyme has a pH optimum of 6.25 and requires Mn2+. The Km values for UDP-GlcNAc and a sugar acceptor substrate were found to be 3.1 mM and 190 microM, respectively. The enzyme activity in the rat kidney was higher than those in the other tissues examined. PMID- 2969204 TI - Control measures against hepatitis B infection. PMID- 2969203 TI - NIH conference. Alzheimer disease: clinical and biological heterogeneity. AB - The clinical and biological features of Alzheimer disease are not uniform in their expression; heterogeneity is evident in the disease's clinical, anatomic, and physiologic characteristics. The presence of considerable intersubject and intrasubject heterogeneity suggests that subtypes of the disease exist. We define subtypes of Alzheimer disease in regard to the behavioral features (for example, predominant right or left hemisphere, or symmetrical impairment), inheritance (familial or sporadic), dosage of chromosome 21 (presence of the Down syndrome), time course of progression, age of onset (presenile or senile), and presence or absence of motor deficit (myoclonus or signs of an extrapyramidal syndrome). Studies of regional cerebral glucose metabolism with positron emission tomography and [18-fluorine] fluorodeoxyglucose show focal alterations in glucose use, with cerebral metabolic asymmetries in patients with Alzheimer disease that are related to the nature of the cognitive deficit. Serial roentgenographic computed tomographic studies show heterogeneous rates of lateral ventricle enlargement in the disease that are related to rates of cognitive decline. Similar anatomic and physiologic abnormalities are also found in persons 45 years of age or older who have the Down syndrome. Furthermore, patients with Alzheimer disease who have extrapyramidal dysfunction or myoclonus are a distinct subgroup, with specific abnormalities of central monoamine markers of dopamine metabolism, serotonin metabolism, and the hydroxylation cofactor, biopterin. The concept of subtypes in Alzheimer disease serves as a model with which the interactions of genetic influences with environmental factors can be examined. PMID- 2969205 TI - Maternal-neonatal transmission of hepatitis B virus. Epidemiology and prevention. PMID- 2969206 TI - Developments in immunization against hepatitis B. PMID- 2969208 TI - Systemic thrombolysis and percutaneous transluminal coronary angioplasty (PTCA) in acute myocardial infarction. AB - Intravenous high-dose infusion of streptokinase in acute evolving myocardial infarction is a widely used therapeutic concept with clinically relevant recanalization rates and low complications. In our experience with 150 patients and acute myocardial infarction treated with intravenous streptokinase (1.5 Mio U), 107 (78 p. 100) of 137 patients demonstrated an antegrade perfused infarct artery. In a group of patients (n = 95), in whom early revascularization was performed, the incidence of reinfarction was reduced from 15 p. 100 to 7 p. 100; hospital mortality was not influenced (3.6 p. 100 vs 4.3 p. 100). PTCA was successful in 39 of 48 patients (81 p. 100). The incidence of angiographically determined restenosis amounted to 28 p. 100 (9/32). Patients after successful PTCA without restenosis demonstrated an improvement of left ventricular function in contrast to patients with restenosis or reocclusions. Thus, intravenous streptokinase followed by PTCA presents a clinically practicable and promising method for treatment of acute myocardial infarction. PMID- 2969207 TI - Unstable angina. AB - Patients with unstable angina, defined as resting chest pain associated with transient repolarization changes on the electrocardiogram, represent a high risk subset among the clinical manifestations of ischemic heart disease. Pathogenetic mechanisms include coronary spasm and vasoconstriction, coronary thrombosis and platelet aggregation. Early prognosis is related to the degree of activity of the disease while long-term outcome depends on the extent of the coronary disease and the degree of left ventricular dysfunction. Medical treatment should include the combination of beta-blockers, nitrates and calcium antagonists as well as the use of heparin and aspirin. Despite such an aggressive treatment, attacks of resting chest pain persist in almost 30 per cent of patients. In these cases emergency revascularization may be achieved by either coronary angioplasty or bypass surgery. The latter operation may result in improved survival in patients with impaired left ventricular function and triple vessel disease. PMID- 2969209 TI - [Umbilical metastasis of a carcinoid tumor of the digestive tract]. PMID- 2969211 TI - The second meeting of the Cardiovascular and Interventional Radiological Society of Europe (CIRSE). Porto Cervo, Sardinia, May 25-29, 1987. Proceedings. PMID- 2969210 TI - Modified Gianturco expandable wire stents in experimental and clinical use. PMID- 2969212 TI - Laser angioplasty: newer modalities. PMID- 2969213 TI - Recanalization of occluded peripheral vessels by laser assisted PTA. PMID- 2969214 TI - First own results with dynamic angioplasty with the Kensey-catheter. PMID- 2969215 TI - Results and complications of renal PTA. PMID- 2969217 TI - Percutaneous transluminal coronary angioplasty. Ten years of experience. PMID- 2969216 TI - Percutaneous transluminal angioplasty of subclavian artery. Comparative study with axillo-contralateral bypass. PMID- 2969218 TI - Spectinomycin disk zone diameter as a predictor of outcome in clinical treatment of gonorrhea. AB - The MICs for 41 Neisseria gonorrhoeae strains from patients receiving spectinomycin treatment were determined by the agar dilution method and compared with the zones of inhibition produced by disks containing 100 micrograms of spectinomycin. Our data demonstrated a good correlation between the two methods. Moreover, a zone of inhibition of less than or equal to 15 mm was a good predictor of clinical treatment failures with spectinomycin. PMID- 2969219 TI - Epidermis and lymphocyte interactions during a tuberculin skin reaction. I. Increased ETAF/IL-1 like activity, expression of tissue antigens and mixed skin lymphocyte reactivity. AB - Forty-one persons were tested for tuberculin skin reactivity. Epidermal cells (ECs) were isolated from the tuberculin reaction and from a contra lateral, non injected skin area. We found a significant increase of epidermal thymocyte activating factor (ETAF) in epidermis overlying a positive tuberculin reaction together with an increase of OKT6 and class II (HLA-DR) positive cells. Allogeneic lymphocytes proliferated significantly more when mixed with ECs from a positive tuberculin skin test. Injection of tuberculin per se or a negative reaction did not induce similar changes. The described model seems useful for functional studies of ECs and lymphocytes in patients with contact dermatitis. PMID- 2969220 TI - Influence of oral isotretinoin treatment on the composition of comedonal lipids. Implications for comedogenesis in acne vulgaris. AB - One of the primary events in the pathogenesis of acne vulgaris is abnormal follicular keratinization. Since oral isotretinoin therapy reduces follicular hyperkeratinization in acne, our study has been designed to determine whether epidermal lipid composition of the epithelium of sebaceous follicles is affected by isotretinoin treatment. Noninflamed early comedones obtained from ten patients with nodulocystic acne before and after the 6th week of isotretinoin therapy (mean daily dose 0.7 mg/kg b. wt.) were used as probes of the hyperkeratinizing follicular epithelium. Comedonal lipids were analyzed by high-performance thin layer chromatography. Oral isotretinoin caused a decrease of the comedonal glyceride fraction by 36% (P less than 0.01), whereas free sterols and total ceramides increased by 34% (P less than 0.10) and 19%, respectively. The changes of comedonal lipids were associated with a significant elevation of the free sterols/cholesterol sulfate ratio of 86% from pretreatment levels (P less than 0.05). The isotretinoin-induced changes of the comedonal lipid composition in direction to a pattern of epidermal lipids of normal desquamating stratum corneum are discussed as a possible comedolytic mechanism of oral isotretinoin treatment. PMID- 2969221 TI - Influence of chlorpromazine on motility and calcium uptake of boar sperm. AB - In the motility of sperm, which resembles the automaticity of smooth muscle in some features, calcium plays an important role. Chlorpromazine (CPZ), an inhibitor of calmodulin (CaM), decreases contractile responses of smooth muscle to agonists, alters microsomal calcium movements and inhibits human sperm motility. The involvement of CaM in the contraction of smooth muscle and in sperm motility and its presence in sperm cells have also been established. Therefore, the effects of CPZ on boar sperm motility and microsomal calcium uptake were studied. CPZ (0.05-0.5 mM) was found to depress the motility index of boar sperm. At 60 min time point, an inhibition of the motility index ranging from 28 to 85% was observed (p less than 0.05). The spermatozoa microsomal calcium uptake (5.2 nmol/mg protein/hr) was Mg++- and ATP-dependent and azide insensitive. Ionophore A23187 (1 microM) and EDTA (10 mM) inhibited the uptake (67-94%). The calcium uptake was inhibited (40-60%) by CPZ (0.25 and 0.5 mM). The CPZ-induced inhibition of the motility index or calcium uptake was overcome by additional CaCl2. The observations suggested that the CPZ-caused changes were calcium mediated and possibly involved CaM. PMID- 2969222 TI - Influence of the chronic administration of LH-RH agonist on the oral glucose tolerance test in patients with prostatic carcinoma. PMID- 2969223 TI - [A new method of laser angioplasty by contact sapphire: preliminary results. Apropos of 20 cases]. AB - We report our first 20 cases of peripheral laser angioplasty using an optic fibre with contact sapphire tip. The equipment included a teflon catheter on which was screwed a round sapphire 2.2 mm in diameter. A 600 microns optic fibre connected to a Nd-Yag laser instrument was introduced into the catheter and placed in contact with the sapphire. Twenty patients underwent recanalization of femoral or popliteal arteries occluded on a length of 5 to 45 cm. The sapphire-tipped catheter was introduced by the Seldinger technique up to the site of occlusion. The 15 watt laser emission was set at intervals of one second. Sixteen out of the 20 occluded arteries were recanalized. Among the 4 failures, 3 were due to perforation and 1 to intraparietal progression. Angioplasty was performed with laser alone in 3 cases and with laser completed by balloon catheter in 13 cases. The minimum diameter of the laser-induced channel was 2 mm and was significantly increased (3.8 mm) by complementary balloon dilatation. In the 3 patients who underwent laser angioplasty alone, no noticeable improvement in distal blood flow was demonstrated by doppler velocimetry, and reocclusion occurred either soon afterwards (n = 2) or later (n = 1). Midterm results were much better in patients who had had additional balloon dilatation: early (3rd day) or late (2 months) reocclusion took place in only 3 patients. In the remaining 10 patients, followed up for periods of 1 week to 18 months (mean: 6 months), clinical improvement and recanalization were maintained. PMID- 2969224 TI - [Contribution of excimer laser to coronary angioplasty]. AB - Reports on the problems encountered with thermal laser system in angioplasty have prompted us to test an excimer Xe-Cl laser of 308 nm wavelength and 20 ns impulses on preparations of healthy and atheromatous coronary arteries, the purpose of the study being to quantify the adequate parameters required for angioplasty. We effectively used optic fibres of different diameters--a method which has not yet been reported in medical literature. Our choice of an ultraviolet laser was justified on theoretical grounds: essentially its mode of photo-ablation devoid of thermal effects and proportional to the number of inter- and intramolecular bonds destroyed. The photo-ablation threshold we measured being 1.40 J/cm2, we micrometrically determined the volume of matter removed by various combinations of energy intensities and numbers of impulses, as well as the undesirable effects of such combinations. We found that with an energy surface density of 4.23 J/cm2 the depth of ablation was directly proportional to the number of impulses (between 20 and 100). Under these experimental conditions, the ablation produced by the Xe-Cl laser system was 14 microns per impulse in depth, and of the same diameter as that of the optic fibre used. Virtually identical data were obtained in healthy, fibrotic, atheromatous and calcified tissues. Undesirable effects, i.e. coagulation necrosis, only appeared with high intensity energies irrelevant to the study. No lesion of the parietal cells was observed, even in the vicinity of the ablation plane. Finally, the question of oncogenic effects raised with 193 nm laser beams does not seem to apply to the 308 nm wavelength.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969225 TI - [Doppler study of phasic variations of blood flow velocity in the anterior interventricular artery]. AB - Studies of changes in coronary blood flow during cardiac cycles may be a useful adjuvant to the measurement of coronary flow reserve to evaluate the hydraulic severity of coronary arterial stenoses. We used intracoronary pulsed Doppler velocimetry to measure phasic variations of blood flow in the anterior interventricular artery of 12 patients with angiographically identified stenosis of that vessel. The Doppler signal was obtained by means of a 20 MHz emission from a source placed at the tip of a catheter selectively positioned at the ostium of the anterior interventricular artery, upstream of the stenosis. The increase in severity of stenosis was paralleled by a relative decrease of diastolic blood flow velocity in relation to systolic blood flow velocity. The diastolic/systolic maximum velocities ratio was greater than 1 in 6 patients with a less than 70 p. 100 stenosis (group A) and inferior to 1 in 6 other patients with a 70 p. 100 or more stenosis (group B). In 5 patients of group B this ratio was reversed to normal after percutaneous transluminal angiography. Thus, measurement of intracoronary blood flow velocity may be helpful to evaluate the severity of stenosis, notably in the anterior interventricular artery where angiographic evaluation is difficult. PMID- 2969226 TI - [Late effect of intracoronary urokinase. Apropos of a case of recurrent coronary thrombosis after angioplasty]. AB - The authors report a case of percutaneous dilatation of a coronary artery performed immediately after a mild myocardial infarction and complicated by occlusive thrombosis without dissection, despite a presumably effective anticoagulant treatment. A second dilatation resulted in rapid recanalization of the artery, but recurrent thrombosis developed at the site of dilatation. The thrombotic process was controlled with an intracoronary infusion of urokinase and higher doses of intravenous heparin, but only after a long delay (80 minutes). This case suggests that in similar circumstances one must wait long enough before referring the patient to a surgical unit for emergency aorto-coronary bypass. PMID- 2969227 TI - [Vertebral osteochondroma in lumbosciatalgia caused by herniated disk. Case report]. PMID- 2969228 TI - Self-concept, sexual knowledge and attitudes, and parental support in the sexual adjustment of women with early- and late-onset physical disability. AB - Three groups of college women were compared on the following dimensions: sexual experiences, sexual satisfaction, self-concept, sexual attitudes and knowledge, and parental support for sexual development. Groups were composed of women with late-onset physical disability, early-onset physical disability, and no disability. Measures utilized included an extension of the Sexual Interaction Inventory, the Tennessee Self-Concept Inventory, the Sexual Knowledge and Attitude Test, and questionnaires developed for this study. The early-onset group reported fewer current sexual experiences than did the nondisabled group. The early-onset group was dissatisfied with the frequency of sexual behavior to a greater degree than the nondisabled group. Also, both disabled groups believed that they could enjoy sexual experiences more than they did at present in contrast to the nondisabled group. There were no group differences for self concept, though several aspects of self-concept contributed significantly to sexual adjustment for the disabled groups. No group differences were found for sexual knowledge and attitudes or parental support for sexual development, nor did these variables relate to sexual adjustment. Group differences in sexual adjustment are discussed in terms of functional, emotional, and social implications. Recommendations for further research include following the social development of women with late-onset conditions, comparing the social skills and cognitions of the two disabled groups, and assessing men who have established intimate relationships with disabled women. PMID- 2969229 TI - Catheterization and angioplasty of the nonopacified peripheral autogenous vein bypass graft. AB - Nonopacified lower extremity vein bypass grafts may not be thrombosed. Catheterization of these grafts should be performed whenever possible. If graft patency is demonstrated, vein graft angioplasty may restore flow without fibrinolytic therapy. In two cases of nonthrombosed nonopacified grafts, graft patency was demonstrated by catheterization of the grafts, and normal blood flow was restored by percutaneous transluminal angioplasty. PMID- 2969230 TI - Right trisegmentectomy with a synthetic vena cava graft. AB - In the course of a right trisegmentectomy of the liver, nearly the entire length of the retrohepatic vena cava was replaced with a Dacron graft. Patency of the graft was proved by a venacavogram a year later. PMID- 2969231 TI - Surgical closure of atrial septal defect in patients older than 50 years of age. AB - Between 1962 and 1986, 51 patients 50 years of age or older (mean, 59.7 years; range, 50 to 77 years) underwent operative closure of atrial septal defect. The mean pulmonary vascular resistance was 1.7 +/- 0.79 wood units and the mean pulmonary blood flow-systemic blood flow ratio was 2.8 +/- 1.4. Patch closure with pericardium or Dacron was performed in 36 patients (70%), while 15 patients (30%) underwent primary closure. There were no operative deaths, and one patient was not available for follow-up. The remaining 50 patients (98%) have been followed up for 0.5 to 25 years (mean, 9.6 years). Mean preoperative New York Heart Association classification was 2.40 +/- 0.70. This improved significantly to 1.21 +/- 0.42 at follow-up, with all patients improving by at least one category. No patient received long-term anticoagulation treatment, and no pulmonary or systemic emboli were identified. Two (13%) of 15 patients who underwent primary closure developed septal dehiscence. Actuarial survival was 93%, 86%, and 79% at 5, 10, and 15 years, respectively. Atrial septal defect closure can be safely performed in older patients with excellent results, provided the ratio of systemic to pulmonary blood flow is greater than or equal to 1.5 to 1.0, the pulmonary vascular resistance is low, and the shunt remains left to right. Primary closure should be discouraged, and postoperative anticoagulation therapy appears unwarranted. PMID- 2969233 TI - Conditions associated with Huntington's disease at death. A case-control study. AB - To identify conditions associated with reduced survival in patients with Huntington's disease, we studied all 1978 US death certificates on which Huntington's disease (331.0 in the International Classification of Diseases, eighth revision) was listed. For each of the 495 cases identified, two control deaths were matched by age, race, sex, county, and year of death. Pneumonia, choking, nutritional deficiencies, and chronic skin ulcers were increased in cases relative to controls. PMID- 2969232 TI - Thiamine and Alzheimer's disease. A pilot study. AB - As a test of the significance of previously described biochemical abnormalities in thiamine-dependent enzymes in brains and other tissues in patients with Alzheimer's disease, a double-blind, placebo-controlled, crossover, outpatient pilot study compared the effects of 3 g/d of oral thiamine hydrochloride for three months with those of a niacinamide placebo. Eleven moderately impaired patients with "probable Alzheimer's disease" by the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria completed the study. All patients were well nourished and had no stigmata of dietary thiamine deficiency. Their initial mean +/- SEM Mini-Mental State Examination score was 14.2 +/- 1.4, and the mean age was 72 years. Global cognitive rating by the Mini-Mental State Examination was higher during three months with 3 g/d of oral thiamine hydrochloride than with niacinamide placebo. Behavioral ratings, however, did not differ significantly, nor did clinical state when it was judged subjectively. PMID- 2969234 TI - AADS adopts AIDS policy. PMID- 2969236 TI - Long-term study on T lymphocyte subsets in newly diagnosed type 1 diabetes mellitus. AB - T lymphocyte subsets in peripheral blood from 16 newly diagnosed type 1 diabetic children were studied prospectively at four time intervals: as soon as possible after diagnosis and 1, 4 and 12 months later. T lymphocyte subsets were analysed using monoclonal antibodies and counted by cytofluorimetry. The percentage of T lymphocytes (OKT3+ cells) did not change at the four study times. The percentage of helper/inducer T cells (OKT4+ cells) was high at the diagnosis (43.1 +/- 2.1%), but decreased after 1 and 4 months with no difference in the control values. The percentage of suppressor/cytotoxic T lymphocytes (OKT8+ cells) was low at the diagnosis, but increased after 1 and 4 months. The OKT4/OKT8 ratio was 2.31 +/- 0.22 at the diagnosis study, decreasing to 1.83 after 1 month, compared with 16 sex- and age-matched control children. The high percentage of helper/inducer T lymphocytes and low number of suppressor/cytotoxic T cells at onset of diabetes favour immune reactions that lead to beta-cell damage. PMID- 2969235 TI - Lipoprotein Lp(a). A risk factor for myocardial infarction. AB - The aim of this study was to test plasma lipoprotein Lp(a) and other lipid and lipoprotein levels for association with the incidence of myocardial infarction. Total plasma cholesterol, triglycerides, low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, and Lp(a) were measured in 1486 men at the age of 18 years. In addition, the Broca Index (a measure of relative body weight) and other data were recorded. The sample was divided into probands whose mothers or fathers suffered a myocardial infarction (case group, n = 52) and into probands whose parents had no myocardial infarction (control group, n = 1434). In the case group, 32% had Lp(a) plasma concentrations greater than 25 mg/dl, but only 13.4% of the control group had this level of concentration, a highly significant difference (p less than 0.01). In addition, there was a statistically significant difference in the ratio of LDL cholesterol/HDL cholesterol (p less than 0.05) and the Broca Index (p less than 0.01) between cases and controls. The parents of the case group were significantly older than the parents of the control group; however, when a control group was matched for parents' age, the results were similar. These data suggest that parents of male children with Lp(a) plasma concentrations greater than 25 mg/dl have a 2.5-fold higher incidence of myocardial infarction. Considering the familial aggregation of elevated Lp(a) levels, we conclude that increased levels of this lipoprotein may be a significant risk factor for myocardial infarction. PMID- 2969237 TI - ATP-dependent Cl- uptake by plasma membrane vesicles from the rat brain. AB - Uptake of Cl- by plasma membrane vesicles from the rat brain was stimulated by ATP at 37 degrees C, but not by beta, gamma-methylene ATP or at 0 degrees C. The addition of Triton X-100 or sucrose to the incubation medium diminished the ATP stimulated Cl- uptake, suggesting that Cl- was transported across the membranes into the intravesicular space. This ATP-stimulated Cl- uptake was not affected by 1 mM ouabain. 1 microM oligomycin, 0.1 mM gamma-aminobutyric acid or 0.1 mM picrotoxin. Thus, non-mitochondrial ATP-driven Cl- transport through a system other than Na, K-ATPase or Cl- channels occurs in neuronal plasma membrane vesicles. PMID- 2969238 TI - Distribution of receptors for insulin and insulin-like growth factor I (somatomedin C) in the adrenal gland. AB - Rat adrenal glands contain cell surface high-affinity receptors for several peptide hormones. Receptors for IGF-I were abundant in this tissue, but receptors for insulin were relatively scarce. The behavior of adrenal membrane IGF-I receptors in radioligand binding assays was similar to the behavior of IGF-I receptors from other tissues, with a KD congruent to 6.2 x 10(-9) M. Covalent cross-linking studies with [125I]IGF-I revealed an IGF-I receptor alpha-subunit with Mr congruent to 135,000 on dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions, as well as a smaller radiolabeled peptide, Mr = 116,000. In contrast, little binding of [125I]insulin to adrenal membranes was observed and no labeling occurred in cross-linking studies using [125I]insulin. These results contrast with the findings of whole-body autoradiographic studies that indicated substantial binding of [125I]insulin to adrenal glands and suggest that IGF-I, rather than insulin, may play a critical role in the growth and development of the adrenal gland. PMID- 2969239 TI - Characterization of insulin-like growth factor I (IGF-I) receptors of human breast cancer cells. AB - Studies of binding of IGF-I to a plasma-membrane-enriched subcellular fraction prepared from MCF-7 human breast cancer cells reveal the presence of 0.2 pmols specific binding sites for this mitogen per mg membrane protein, with an equilibrium affinity constant of 1.45 nM-1. Competition studies with insulin, IGF II, and an anti-IGF-I receptor antibody are consistent with the presence of specific IGF-I receptors, and SDS-PAGE showed binding to a 130 kDa subunit identical to that of receptors from human placenta. In addition, we show that IGF I is more potent than estradiol and comparable to EGF in stimulating in vitro proliferation of MCF-7 cells, and that IGF-I-stimulated proliferation of these cells is inhibited by a blocking monoclonal antibody against the IGF-I receptor. These results demonstrate that IGF-I is an important mitogen for MCF-7 cells and that the mitogenic effect is mediated by specific IGF-I receptors. PMID- 2969240 TI - Chlorate: a reversible inhibitor of proteoglycan sulfation. AB - Bovine aorta endothelial cells were cultured in medium containing [3H]glucosamine, [35S]sulfate, and various concentrations of chlorate. Cell growth was not affected by 10 mM chlorate, while 30 mM chlorate had a slight inhibitory effect. Chlorate concentrations greater than 10 mM resulted in significant undersulfation of chondroitin. With 30 mM chlorate, sulfation of chondroitin was reduced to 10% and heparan to 35% of controls, but [3H]glucosamine incorporation on a per cell basis did not appear to be inhibited. Removal of chlorate from the culture medium of cells resulted in the rapid resumption of sulfation. PMID- 2969241 TI - Atriopeptin III depresses the excitability of sympathetic neurones. AB - The effect of atriopeptin III (AP III) on the excitability of the inferior mesenteric ganglion (IMG) of the guinea-pig was studied by intracellular recording technique. AP III depressed the excitability of the IMG by decreasing the frequency of the action potentials, prolonging the afterpotential and increasing the threshold of the action potentials. Voltage-clamp studies revealed that a transient potassium current-the A current-was enhanced by AP III. It is suggested that inhibition of the sympathetic nervous system may contribute to the cardiovascular effect of AP III. PMID- 2969242 TI - Isolation and characterization of non-neuronal enolase (NNE) from Neurospora crassa and comparison with neuron specific enolase isolated from neuroblastoma cell line NG108. AB - Enolase is a vital enzyme of the glycolytic pathway. It exists mainly in two forms, non-neuronal enolase (NNE) and neuron specific enolase (NSE). Neurospora crassa, a filamentous fungus, was used as the source of pure NNE, and by using DEAE-cellulose and a Sephadex G-150 column chromatography highly purified enzyme (20.4 fold purification with 54.7 percent recovery) was obtained. The development profile of the enzyme shows a peak value after 90 hours of mycelial growth from conidia of N. crassa. In this respect, it differs from neuroblastoma NSE where the peak value of the enzyme activity appears 7 1/2 hours after the splitting of the cells. N. crassa enolase (NNE) is more thermolabile than NG108 NSE and N. crassa enolase is more sensitive to urea, chloride, and fluorophosphate. The Km values for 2-phosphoglycerate and Mg++ were 0.34 mM and 0.47 mM, respectively, for N. crassa enolase, whereas these values were 1.1 mM and 3.1 mM, respectively, in the case of neuroblastoma NSE. N. crassa enolase is a dimer molecule of molecular weight 85,000 daltons. N. crassa enolase is not neutralized by NSE antisera and neutralized by NNE antisera as opposed to neuroblastoma NSE. PMID- 2969243 TI - Identification and characteristics of a novel mitochondrial ATPase in rat liver. AB - A novel ATPase is postulated for isolated mitochondria and mitoblasts of rat liver. The enzyme is active in the presence of oligomycin and carboxyatractyloside. It can be distinguished from other well-known mitochondrial and non-mitochondrial ATPases by its insensitivity to common ATPase inhibitors and effectors and by digitonin treatment. The ATPase is localized on the outer side of the inner mitochondrial membrane. It is activated by Mg2+ in the alkaline pH range and exhibits a biphasic kinetics. The novel external ATPase of rat liver mitochondria possesses similar properties with respect to ATP-dependent protease. PMID- 2969244 TI - Comparative toxicity of 4-chlorobiphenyl and its metabolite 4-chloro-4' biphenylol in isolated rat liver mitochondria. AB - 4-Chlorobiphenyl (4-CB) is converted by the microsomal cytochrome P-450 system to its hydroxylated metabolite 4-chloro-4'-biphenylol (4'-OH-4-CB). A study of the effects of 4-CB and 4'-OH-4-CB on the energy-linked functions of rat liver mitochondria was carried out. 4'-OH-4-CB was more effective than 4-CB in causing the inhibition of state 3 respiration of mitochondria with both succinate and glutamate/malate. As a substrate specificity, with glutamate/malate the inhibition by each compound (ID50, 30 microM for 4'-OH-4-CB, 76 microM for 4.CB) was more significant than that with succinate (ID50, 200 microM for 4'-OH-4-CB, never reached 50% for 4-CB). From the effects on DNP-stimulated respiration, it was indicated that the electron transport from both glutamate/malate and succinate to oxygen was more sensitively inhibited by 4'-OH-4-CB than by 4-CB, with the same substrate specificity as for state 3 respiration (i.e. the inhibition by both compounds was greater with glutamate/malate than with succinate). Since there existed a good coincidence in the inhibition between state 3 and DNP-stimulated respiration with both substrates, the inhibition of state 3 respiration by both compounds was due to the inhibition of the electron transport. With succinate, the uncoupling of oxidative phosphorylation by both compounds was observed, the extent of which was greater with 4'-OH-4-CB than with 4-CB, although the uncoupling by higher concentrations of 4'-OH-4-CB was masked because of the increased inhibition in respiration. With glutamate/malate, the uncoupling action of 4-CB was largely, while that of 4'-OH-4-CB was completely, masked by progressive respiratory inhibition. 4'-OH-4-CB was more effective than 4-CB in causing stimulation of latent ATPase in mitochondria. These results indicate that both 4-CB and 4'-OH-4-CB impair mitochondrial energy-transducing functions, but 4'-OH-4-CB is more effective than 4-CB in damaging these functions. Thus, the product of the metabolism is more biologically active than the parent compound. The impairment of energy-linked mitochondrial reactions by the metabolite as well as of the parent compound may be an important factor in the toxicity of 4-CB. PMID- 2969245 TI - Aspects of cytokine induced modulation of immunity and inflammation with emphasis on interleukin 1. AB - Interleukin 1 (IL 1) contributes to both acute and chronic inflammatory processes. Purified IL 1 by itself is only one of the signals that induces local inflammation. The contribution of IL 1 to local inflammatory reactions is influenced by exogenous stimuli, antagonists such as steroid hormones and TGF beta (tissue growth factor beta) and agonists such as TNF (tissue necrosis factor). IL 1 production is depressed by glucocorticoids, but glucocorticoids induce expression of receptors for IL 1 on selected cell types including B lymphocytes and fibroblasts. Thus, complex interactions between hormones and cytokines modulate immunological and inflammatory responses. PMID- 2969246 TI - The role of protein C and protein S in the haemostatic process. AB - Protein C and Protein S are vitamin K dependent clotting factors. The importance of these proteins in the haemostatic process was recognized when deficiencies of Protein C and Protein S were detected that were associated with thromboembolic disorders. In this review the function of Protein C and Protein S in the haemostatic process is described. PMID- 2969247 TI - [Atrial natriuretic peptide in cardiac insufficiency]. AB - The role of atrial natriuretic peptide in the pathophysiology of heart failure is unknown. The aim of the study were changes of atrial natriuretic peptide, hemodynamic, renal and hormonal parameters during the development of cardiac failure in an animal model of congestive heart failure in the conscious dog due to rapid right ventricular pacing and in rats with chronic left ventricular failure due to a left ventricular infarction. The effects of intravenous administration of atrial natriuretic peptide were studied in patients with severe congestive heart failure, dogs with experimental cardiomyopathy and conscious rats with acute right ventricular failure due to repeated pulmonary emboli. The results suggest an important role of atrial natriuretic peptide in the early phase of heart failure as a counterregulating system concerning vasoconstrictory and volume retaining mechanisms like the renin-angiotensin-aldosterone system, the sympathetic nerve activity and vasopressin. In chronic heart failure the renal effects of atrial natriuretic peptide are attenuated. Pharmacological doses have beneficial effects on ventricular function by reducing pre- and afterload. The reduction in effectiveness of atrial natriuretic peptide in congestive heart failure may be due to a down-regulation of specific receptors, or caused by hemodynamic renal changes preventing the action of the hormone on the kidney in heart failure or may be due to an activation of counterregulating systems overridding the effects of atrial natriuretic peptide. PMID- 2969248 TI - [Lack of T lymphocytes expressing CD4 antigen during pollenosis]. AB - T lymphocyte subpopulations by monoclonal antibodies have been studied in 17 atopic asthmatics and 17 normal subjects age and sex matched, we found a significant decrease of total T lymphocytes with significant decrease of CD4 antigen bearing cells in the patients, the pollenogenic asthmatics being responsible for this difference with the controls. We did not find any study of the literature evaluating T lymphocyte subpopulations in atopic patients with regard to allergen type. The pathogenic hypothesis are discussed. PMID- 2969249 TI - [Type III (Arthus) reaction during desensitization]. AB - We rapport an original observation of adverse reaction (Arthus reaction) during a dust mite hyposensitisation with a aluminum hydroxide absorbed extract. We precise the way of beginning. In discussion, we try to precise the preliminary symptoms of such reaction to prevent it. PMID- 2969250 TI - Beta-endorphin genetics in the etiology of alcoholism. AB - "Reinforcing" effects are ascribed to endogenous opioids, particularly to the pro opiomelanocortin (POMC)-derived beta-endorphin 1-31, the most potent opiate active substance. Alcohol induces variations in the genetic processing of the precursor POMC and of beta-endorphin at different levels. Studies focused on changes in POMC gene expression (mRNA quantitation) and post-translational processing. Chronic alcohol intake significantly reduces POMC mRNA in the lobes of the pituitary. In inbred strains of mice, genotypic differences are seen in post-translational processing of hypothalamic beta-endorphin, thus inducing differences in alcohol sensitivity. Clinical studies show a disproportion of POMC cleavage products in the CSF of chronic alcoholics (reduced beta-endorphin versus increased ACTH contents), together with remarkable indications for baseline differences in beta-endorphin levels. Errors within the genetic sequence of POMC are suggested to underlie alcohol-seeking behavior. PMID- 2969251 TI - Solubility behavior of enzymes after addition of polyethylene glycol to erythrocyte hemolysates. AB - The addition of polyethylene glycol to a hemolysate of rat erythrocytes reduces the solubility of phosphofructokinase and glucose-6-phosphate and 6 phosphogluconate dehydrogenases in an exponential manner with respect to polymer concentration. Analyses of the solubility curves (log solubility versus polymer concentration) obtained at different pH values suggest that the solubility can be related to both the aggregation state and the intrinsic solubility of the proteins promoted by solution conditions. These findings suggest the possibility of using polyethylene glycol in a rational way for the fractional precipitation of a mixture. PMID- 2969252 TI - Recirculation: review, techniques for measurement and ability to predict hemoaccess stenosis before and after angioplasty. AB - The measurement of recirculation during two-needle hemodialysis has been shown to provide valuable information concerning distal hemoaccess stenosis and the loss of effective dialysis. Reference sources have suggested that levels of recirculation above 20% call for further investigation. We proposed a technique for routine recirculation measurement and studied 20 stable hemodialysis patients using this method. We found that all patients had recirculations of less than 10%. We also studied 2 patients with angiographically proven stenosis and found 1 patient with a recirculation of less than 20%. Routine recirculation testing provides valuable information about hemoaccess integrity and using our techniques, recirculation greater than 10% is an indication for further study. PMID- 2969253 TI - Echocardiographic determination of myocardial mass: a review. AB - Studies have shown that hypertensive patients with increased left ventricular wall mass are at higher risk for cardiovascular complications than are patients with normal left ventricular wall mass. Echocardiographic determination of myocardial mass is a more sensitive predictor of left ventricular hypertrophy (LVH) than are electrocardiographic or chest X-ray studies. Two-dimensional echocardiographic methods used to determine left ventricular mass have been shown to be a sensitive indicator of assessing left ventricular muscle mass. Left ventricular mass estimates can be used to determine prognosis and cardiovascular risk and to assess the efficacy of therapy in a number of different heart diseases in which LVH is present. PMID- 2969254 TI - Raised concentrations of glucose and adrenaline and increased in vivo platelet activation after myocardial infarction. AB - Plasma concentration of beta thromboglobulin was used as an index of in vivo platelet activation in 36 patients after acute myocardial infarction. Twelve patients had diabetes, seven had pulmonary oedema or cardiogenic shock (pump failure) or both, and 17 had uncomplicated infarcts. On the first day of admission, concentrations of beta thromboglobulin were higher in the patients with diabetes and those with pump failure than in those with uncomplicated infarcts. Concentrations of beta thromboglobulin in the non-diabetic patients were studied by multiple regression analysis and were significantly associated with plasma concentrations of adrenaline, pump failure, and glucose but not with noradrenaline or infarct size. When all subjects were considered together, glucose, adrenaline, and pump failure were associated with the beta thromboglobulin concentration but diabetes was without significant effect. Hyperglycaemia and raised plasma adrenaline concentration after myocardial infarction may activate platelets, and this could contribute to poor outcome in such patients. PMID- 2969255 TI - In vivo emergence of a highly metastatic tumour cell line from a rat rhabdomyosarcoma after treatment with an alkylating agent. AB - Rats bearing a transplanted nickel-induced rhabdomyosarcoma (RMS 9-4/0), treated with chlorozotocin (CZT), an alkylating agent, showed an amplified metastatic invasion of the lung (median of 165 lung tumour nodules, compared to 3 for untreated controls). A higher level of metastatic invasion (200 nodules) was reached spontaneously after the grafting of the S4T line, which was obtained by successive in vivo passages of RMS 9-4/0 cells in CZT treated rats. S4T tumour cells also invaded the liver and a considerable proportion of the lymph nodes. The NT4T line, obtained by successive in vivo passages in untreated rats, showed a lesser degree of enhancement of metastatic capacity (57 nodules). Both derived lines proved to be more aggressive than the parental, proliferated more rapidly, and were resistant to CZT toxicity. Only the non-treated lineage became more resistant to NK lysis. The S4T line lost its myogenic differentiation and was best described as a fibrohistiosarcoma, whereas NT4T did not. Chromosome analysis demonstrated a reduced range of chromosome number per cell in both lines. We conclude that both S4T and NT4T tumours became more metastatic than RMS 9-4/0 as the result of tumour progression through in vivo passages, and that in addition S4T acquired a spontaneously higher metastatic potential, similar to that which occurred in rats grafted with RMS 9-4/0 or NT4T tumours and treated by CZT. This suggests an inheritable mutation in the S4T line. PMID- 2969256 TI - A histological and immunocytochemical study of early acne lesions. AB - We examined 69 biopsies of acne lesions known to be 6, 24 or 72 h old and found that the lymphocyte was the predominant cell at 6 and 24 h. The helper:suppressor T cell ratio in the inflammatory infiltrate was 2.8:1. Polymorphonuclear leukocytes were increasingly seen at 24 and 72 h and were associated with disruption of the duct. We hypothesize that this early lymphocytic infiltrate represents a cell mediated immune response to an antigen within the duct lumen. PMID- 2969257 TI - The effect of simple warming procedures on finger blood flow in systemic sclerosis. AB - Finger blood flow was measured by laser Doppler flowmetry in 15 patients with Raynaud's phenomenon (RP) due to systemic sclerosis (SS), and 15 normal controls. Measurements were performed in a temperature controlled room at 28 degrees C. The blood flow in the patients was significantly lower than in the controls (P less than 0.001). After hand warming in water at 35 degrees C for 10 min, blood flow in the patients and controls did not differ significantly. Following this, the response to a standardized cold stress produced similar falls in both groups to levels that were not significantly different and these occurred over a similar time course. After cold stress ended, there was recovery of blood flow in both groups, and blood flow after 20 min was not significantly different between the two groups. The induced vasodilatation persisted, in those patients in whom it was remeasured, for at least 2 h. Repeating the experiments at a room temperature of 24 degrees C produced similar results. This indicates that considerable vasodilatation is possible in these patients, and can be produced by simple means. It also indicates that local and central thermoregulatory reflexes are intact. Cold induced symptoms in patients with SS are related to low resting blood flow, not to cold sensitivity, and simple warming may provide a useful treatment. PMID- 2969258 TI - Effect of nicotinamide on the phototest reaction in polymorphous light eruption. AB - The effect of nicotinamide on the phototest response was investigated in 14 patients suffering from polymorphous light eruption (PMLE). In contrast to the favourable effect in the prevention of PMLE reported by other groups, oral nicotinamide did not affect the phototesting results in our patients. PMID- 2969259 TI - Tolerance of spironolactone. AB - A survey of 54 patients taking spironolactone for hirsutes or acne showed that side-effects occurred in 91%; in 80% of patients, these were related to the anti androgenic mechanism of the drug (menstrual disturbances, and breast enlargement and tenderness). The concomitant use of a contraceptive pill gave a lower incidence of menstrual abnormalities. Only seven patients (13%) had to stop the drug. In a further eight patients, a reduction in dose to between 125 and 175 mg daily achieved a compromise of controlling the disease and the side-effects. Side effects tended to occur early and so regular review during the initial 3 months of treatment is advised. Two patients in our study developed 'chloasma'--a previously unreported complication of spironolactone. This was the only side effect which occurred late in treatment. PMID- 2969260 TI - High dose nicotinamide in the treatment of necrobiosis lipoidica. AB - An open study of high dose nicotinamide in the treatment of 15 patients with necrobiosis lipoidica is reported. Of 13 patients who remained on treatment for more than 1 month, eight improved. Improvement took the form of a decrease in pain and soreness, a decrease in erythema and the healing of ulcers if present, although the skin did not return completely to normal in any patient. There were no significant side-effects, particularly with respect to diabetic control, an important finding as lesions tended to relapse if treatment was stopped. PMID- 2969261 TI - Delayed wound healing and keloid formation following argon laser treatment or dermabrasion during isotretinoin treatment. AB - We report the observation of delayed wound healing and keloid formation in three patients, following dermabrasion or Argon laser treatment administered while they were receiving isotretinoin for acne or rosacea. PMID- 2969262 TI - Back pain, back abnormalities, and competing medical, psychological, and social factors as predictors of sick leave, early retirement, unemployment, labour turnover and mortality: a 22 year follow up of male employees in a Swedish pulp and paper company. AB - A total of 391 male employees in a Swedish pulp and paper company were followed up for 22 years. As a part of a health examination in 1961 back pain reported by the subjects and abnormalities of the back as judged by the physicians were investigated with respect to predictive power regarding sick leave, early retirement, unemployment, labour turnover, and mortality during the follow up period. Univariate analysis showed that abnormalities of the back were better than back pain for predicting early retirement. Neither back pain nor back abnormalities had any predictive power with respect to long term sick leave, labour turnover, or mortality. Multivariate analyses of 26 variables were performed. Both self assessment of general health and back abnormalities were predictive for early retirement with a diagnosis of back disorder on the retirement certificate but self assessed health was a stronger predictor. Age, smoking, and neuroticism were predictors regarding early retirement for all diagnoses. Education had a negative association; neuroticism did not predict early retirement with a diagnosis of back disorder. PMID- 2969263 TI - Secondary processing of neurohormones: intracellular proteolytic cleavage of beta endorphin generates new active neuropeptides. AB - The following short review considers the physiological significance of naturally occurring peptide fragments derived from intracellular secondary processing of beta-endorphin (beta-EP). Particular attention will be given to the question of whether truncated and/or derivatized forms of beta-EP are true end products subject to release and regulation and might serve important new functions. Since collectively endorphins may play a variety of neurobiological roles which may or may not be mediated through analgesic receptors, most of the material reviewed herein deals with the induction of analgesia, the most commonly used measure of the opiate's acute effects. PMID- 2969264 TI - N-acetyl-beta-D-hexosaminidase A from rat urine: partial purification and characterization. AB - N-Acetyl-beta-D-hexosaminidase A was purified from rat urine by ion-exchange chromatography on DEAE-cellulose, followed by concanavalin A chromatography, and finally by chromatography on 2-acetamido-N-(epsilon-aminocaproyl)-2-deoxy-beta glucosylamine-Se pharose 4B. The enzyme was purified 482-fold with a yield of about 7%. The optimal pH was 4.5 for N-acetyl-glucosaminidase activity and 4.0 4.5 for N-acetylgalactosaminidase activity. The enzyme was heat-labile and stable from pH 4.5 to pH 7.0 but it was very unstable at lower pH values. Km values were 0.55 mM and 0.059 mM, respectively. The glycoprotein nature of the enzyme was deduced from its behavior on concanavalin A. The effect of some carbohydrates and ionic compounds on the activities of the enzyme was studied. When N-acetyl-D glucosaminolactone and N-acetyl-D-galactosaminolactone were used as inhibitors, Ki values were also calculated. PMID- 2969265 TI - Effects of pH on contraction of rabbit fast and slow skeletal muscle fibers. AB - We have investigated (a) effects of varying proton concentration on force and shortening velocity of glycerinated muscle fibers, (b) differences between these effects on fibers from psoas (fast) and soleus (slow) muscles, possibly due to differences in the actomyosin ATPase kinetic cycles, and (c) whether changes in intracellular pH explain altered contractility typically associated with prolonged excitation of fast, glycolytic muscle. The pH range was chosen to cover the physiological pH range (6.0-7.5) as well as pH 8.0, which has often been used for in vitro measurements of myosin ATPase activity. Steady-state isometric force increased monotonically (by about threefold) as pH was increased from pH 6.0; force in soleus (slow) fibers was less affected by pH than in psoas (fast) fibers. For both fiber types, the velocity of unloaded shortening was maximum near resting intracellular pH in vivo and was decreased at acid pH (by about one half). At pH 6.0, force increased when the pH buffer concentration was decreased from 100 mM, as predicted by inadequate pH buffering and pH heterogeneity in the fiber. This heterogeneity was modeled by net proton consumption within the fiber, due to production by the actomyosin ATPase coupled to consumption by the creatine kinase reaction, with replenishment by diffusion of protons in equilibrium with a mobile buffer. Lactate anion had little mechanical effect. Inorganic phosphate (15 mM total) had an additive effect of depressing force that was similar at pH 7.1 and 6.0. By directly affecting the actomyosin interaction, decreased pH is at least partly responsible for the observed decreases in force and velocity in stimulated muscle with sufficient glycolytic capacity to decrease pH. PMID- 2969266 TI - A study of the renal actions of amlodipine in the normotensive and spontaneously hypertensive rat. AB - 1. Normotensive Sprague-Dawley and spontaneously hypertensive rats anaesthetized with sodium pentobarbitone were used to determine the systemic and renal actions of amlodipine, a new calcium channel blocking drug. 2. Amlodipine, 200 micrograms kg-1 plus 50 micrograms kg-1 h-1, decreased blood pressure by 12 +/- 3 mmHg in normotensive rats, although the fall was not statistically significant in the hypertensive rats; did not change renal haemodynamics and caused significant increases in urine flow, absolute and fractional sodium excretions of 70%, 91% and 113%, respectively, in normotensive rats and 65%, 91% and 96%, respectively in hypertensive rats. Fractional lithium excretion was unchanged in the normotensive rats but increased by 28% in the hypertensive animals while absolute fluid reabsorption in the proximal tubule did not change in either group. Absolute water and sodium reabsorption in the segments beyond the proximal tubule were unchanged in the normotensive rats but increased in the hypertensive animals by 24% and 22%, respectively, while fractional sodium excretion in this portion of the nephron increased by 88% and 51% in the normotensive and hypertensive rats, respectively. 3. Amlodipine, 400 micrograms kg-1 plus 100 micrograms kg-1 h 1, decreased blood pressure by 12 +/- 4 mmHg in the normotensive and by 27 +/- 5 mmHg in the hypertensive rats. Renal blood flow was not changed in either group of rats and glomerular filtration rate increased by 25% in the spontaneously hypertensive animals. There were significant increases in urine flow, absolute and fractional sodium excretions of 105%, 145% and 142%, respectively, in the normotensive rats and 224%, 421% and 259%, respectively, in the hypertensive rats. Renal blood flow was not changed in either group of rats and glomerular filtration rate increased by 25% in the spontaneously hypertensive animals. There were significant increases in urine flow, absolute and fractional sodium excretions of 105%, 145% and 142%, respectively, in the normotensive rats and 224%, 421% and 259%, respectively, in the hypertensive rats. Fractional lithium excretion was elevated by 29% and 38%, in the normotensive and hypertensive rats, respectively, but absolute fluid reabsorption at the proximal tubule remained unchanged. At the same time there were significant increases in absolute water and sodium reabsorption beyond the proximal tubule of 26% and 18%, respectively, in the normotensive animals and of 63% and 60%, respectively, in the hypertensive animals. Fractional excretion of water and sodium in the nephron regions after the proximal tubule were increased by 55% and 88%, respectively, in the normotensive rats and by 84% and 121%, respectively, in the hypertensive rats. 4. These doses of amlodipine caused modest reductions in blood pressure, minimal changes in renal haemodynamics and a natriuresis and diuresis. Proximal sodium and water reabsorption was not affected by the drug and it is suggested that the changes in tubular fluid handling were compatible with depression of reabsorption further along the tubule. PMID- 2969267 TI - Pharmacological properties of GR38032F, a novel antagonist at 5-HT3 receptors. AB - 1. This paper describes the pharmacology of the novel 5-hydroxytryptamine3 (5 HT3) receptor antagonist GR38032F. 2. On the isolated vagus nerve and superior cervical ganglion of the rat, R,S-GR38032F behaved as a reversible competitive antagonist of 5-HT-induced depolarization with pKB values of 8.61 +/- 0.08 (n = 19) and 8.13 +/- 0.07 (n = 16), respectively. The resolved R- and S-isomers of GR38032F were approximately equipotent as 5-HT antagonists on the rat vagus nerve: the pKB values were 8.95 +/- 0.05 (n = 16) and 8.63 +/- 0.08 (n = 17), respectively. R,S-GR38032F was also an effective antagonist of 5-HT on the rabbit isolated vagus nerve: in this case the pKB value was 9.40 +/- 0.14 (n = 4). 3. On the rabbit isolated heart, low concentrations of R,S-GR38032F (3 X 10(-11)-1 X 10(-9) M) antagonized the positive chronotropic effect of 5-HT and 2-methyl-5 hydroxytryptamine (2-methyl-5-HT). However, the effects of the compound did not appear consistent with simple reversible competition. 4. On the longitudinal smooth muscle of the guinea-pig ileum, R,S-GR38032F caused concentration dependent parallel rightward displacement of the 2-methyl-5-HT concentration contraction response curve; in contrast, a portion of the response to 5-HT appeared resistant to R,S-GR38032F. pKB values estimated from the effects of the compound against 2-methyl-5-HT or the inhibitable portion of the response to 5-HT were 7.31 +/- 0.06 (n = 8) and 7.33 +/- 0.13 (n = 8), respectively. Against 2 methyl-5-HT, R-GR38032F seemed more potent (pKB 7.20 +/- 0.10; n = 6) than S GR38032F (pKB 6.30 +/- 0.05; n = 6). 5. R,S-GR38032F is highly selective for 5 HT3 receptors, and at concentrations of 3 X 10(-6)-3 X 10(-5) M, had negligible agonist or antagonist activity on other 5-HT or non-5-HT receptor-containing tissues on which it was tested. 6. The potency and duration of action of R,S GR38032F in blocking 5-HT3 receptors in vivo were assessed by measuring its ability to antagonize the bradycardic response to 5-HT or 2-methyl-5-HT administered intravenously (i.v.) to anaesthetized animals. For i.v. administration to the rat, the ED50 for R,S-GR38032F against 2-methyl-5-HT (100pgkg-1) was 0.4 (95% confidence limits 0.18- 0.87) ygkg-1 (n = 10); the corresponding value for oral administration to this species was 7.0 (3.0- 22.0)pgkg-' (n = 8-10 per dose level). R,S-GR38032F was similarly effective in the anaesthetized cat. 7. The present results are discussed with reference to the postulated existence of subtypes of the 5-HT3 receptor. PMID- 2969268 TI - Permanent transrectal drainage of a diverticular-related abscess with a double ended pigtail catheter. PMID- 2969269 TI - Effect of LHRH agonist, Zoladex, on ovarian histology. AB - In a phase I clinical trial Zoladex was used as first line hormonal therapy in premenopausal women with advanced breast cancer. Patients on progression of their disease underwent surgical oophorectomy. The histology of ovaries from 23 women treated with Zoladex has been compared with the ovaries from 34 patients who, before the clinical trial of Zoladex, underwent surgical oophorectomy as primary therapy. Both groups show similar follicular phase development. Only 13 per cent of the Zoladex group developed corpora lutea while 58 per cent showed evidence of luteinization in the primary oophorectomy group (P less than 0.01). Non neoplastic follicular cysts were seen more often in the ovaries of the Zoladex treated patients than in the primary oophorectomy group (P less than 0.05). Zoladex appears to arrest not folliculogenesis but follicular maturation with inhibition of ovulation, the follicles subsequently undergoing atresia with follicular cyst formation. PMID- 2969270 TI - Which patients with suspected appendicitis should undergo laparoscopy? PMID- 2969271 TI - Health care of physically handicapped young adults. PMID- 2969272 TI - Benzydamine oral rinse and rash. PMID- 2969273 TI - Airway obstruction and sleep disruption in Down's syndrome. PMID- 2969274 TI - Mental Health Commission defeated over paedophile. PMID- 2969275 TI - Effect of chronic ethanol treatment on dopamine receptor subtypes in rat striatum. AB - Chronic exposure to ethanol (6% in the drinking water, 25 days) reduces the responsiveness of both the dopamine-stimulated and of the dopamine-inhibited adenylate cyclase in rat striatum. The changes in the adenylate cyclase activity are paralleled by alterations in dopamine recognition sites, in fact binding studies using selective ligands indicate that the number of both D1- and D2 receptors is reduced in striatal membranes of treated rats. PMID- 2969276 TI - Mesolimbic dopamine receptor increases two weeks following hippocampal kindling. AB - Kindled seizures developed in rats following repeated electrical stimulation of the left CA1 region of the hippocampus. Two weeks after the final kindled seizure, the densities of dopamine (DA) D2 receptors were assayed in the left and right amygdaloid area, nucleus accumbens, and nucleus caudatus. A significant increase (107%) in the density of DA D2-receptors in the ipsilateral nucleus accumbens occurred. This finding may help to define the long-term neurochemical consequences of kindling. PMID- 2969278 TI - Decreased incidence of neurologic disability among neonates at high risk born between 1975 and 1984 in Alberta. AB - We report the outcome at 2 or 3.5 years of 1463 neonates at high risk born between 1975 and 1984 and cared for in a regional perinatal program in Alberta. Although the number of surviving infants of very low birth weight (1250 g or less) increased over the study period, the incidence rate of neurologic impairment fell significantly, from 19% to 13% (p less than 0.01), so that there was no significant increase in the absolute number of disabled children. This finding remained valid when two other groups of infants at high risk (those weighing more than 1250 g at birth and having a positive neurologic history and those born at term with asphyxial encephalopathy) were included in the analysis, so that over the decade there was a significant decrease in the incidence of disability among the total group of neonates (p less than 0.01) and no increase in the absolute number of disabled children (23 in 1975 and 19 in 1984). We conclude that neonatal intensive care has contributed to improved survival of neonates at high risk without increasing the burden of major neurologic disability. PMID- 2969279 TI - Photodynamic therapy of endobronchial malignancies. AB - Forty-nine tumor sites in 31 consecutive patients with tracheobronchial malignant neoplasms were treated with photodynamic therapy (PDT). After sensitization with the intravenous hematoporphyrin derivative (HpD) or its more purified form of dihematoporphyrin ether (DHE), 630 nm of light from a tunable sensitizer argon light system was delivered to the tumor site through the biopsy channel of a flexible bronchoscope. All patients had received, refused, or were ineligible for conventional surgery, ionizing radiation therapy, or chemotherapy. Before or at 1 month after each treatment, tumor response was evaluated according to the following categories: (1) complete response (CR) (no visible abnormality, and negative biopsy specimen and cytology); (2) partial response (PR) (degree of obstruction or size of tumor reduced more than 50%); (3) some response (SR) (degree of obstruction or size of tumor reduced 20% to 50%); and (4) progression (PROG) (degree of obstruction or size of tumor reduced by less than 20%). Results were as follows: (1) 37% of the tumors treated achieved CR; (2) 55% achieved PR; (3) 4% achieved SR; and (4) 4% were categorized as PROG. Complete follow-up was achieved in all patients. Clinical effect was evaluated 1 month after treatment by comparing the Karnofsky performance status (KPS), dyspnea level, oxygen requirement, and presence of symptoms. Sixty-eight percent had clinical improvement in at least one variable and 48% in two or more variables. The results of this study suggest that PDT can play a useful role in the treatment of endobronchial malignancies. PMID- 2969277 TI - Bisphosphonates inhibit 1,25-dihydroxyvitamin D3-induced increase of osteocalcin in plasma of rats in vivo and in culture medium of rat calvaria in vitro. AB - In order to test whether bisphosphonates, which are potent inhibitors of osteoclastic bone resorption, may also act upon osteoblasts, we studied the effect of dichloromethylenebisphosphonate (Cl2MBP) and 4-amino-1 hydroxybutylidene-1,1-bisphosphonate (AHBuBP) on in vivo levels and in vitro release of osteocalcin, a bone-specific protein produced by osteoblasts. In rats, 161 mumol/kg of Cl2MBP or 1.61 mumol/kg AHBuBP strongly inhibited the increase of plasma osteocalcin induced by 1,25(OH)2D3. The inhibition was measurable within 24 hours after the administration of bisphosphonate and was independent of any change in bone resorption. The effect upon osteocalcin release was also present in calvaria cultures. 250 microM Cl2MBP strongly inhibited the osteocalcin release induced by 10(-8) M 1,25(OH)2D3. In the presence of 1,25(OH)2D3, protein synthesis and DNA synthesis were also decreased, whereas in the absence of 1,25(OH)2D3, protein synthesis was increased. Thus, bisphosphonates affect the production of a bone-specific protein by osteoblasts in addition to their inhibitory action on osteoclasts. PMID- 2969280 TI - Differential cell photosensitivity following porphyrin photodynamic therapy. AB - Experiments were performed to determine if differences in porphyrin photosensitivity could be observed for cells with varying efficiency in DNA damage repair, as well as for cells which make up components of the vasculature. Photofrin II is undergoing current clinical evaluation for photodynamic therapy of solid tumors, and therefore the retention, dark toxicity, and photosensitizing effects of this drug on human DNA repair-deficient fibroblasts (ataxia telangiectasia and xeroderma pigmentosum) were compared to normal human fibroblasts. In addition, bovine cells of endothelial, smooth muscle, and fibroblast origin were compared for porphyrin retention, toxicity, and photosensitivity. All human fibroblasts exhibited porphyrin-induced dark toxicity, but there were no significant differences in photosensitization or porphyrin retention for any of these cell lines. However, bovine endothelial cells were considerably more photosensitive than smooth muscle or fibroblast cells treated under identical conditions. All bovine cells accumulated similar levels of porphyrin, and therefore the increased sensitivity of the endothelial cells was not due to differences in porphyrin retention. These results provide additional evidence that nuclear damage and/or repair is not a dominant factor in the cytotoxicity induced by porphyrin photosensitization. In addition, these results indicate that endothelial cell photosensitivity may play a role in the vascular damage observed following photodynamic therapy. PMID- 2969281 TI - Development and characterization of a rat model for locally recurring mammary tumors: sensitivities to 5-fluoro-2'-deoxyuridine, adriamycin, and X-irradiation. AB - Local recurrence occurs in 4-47% of breast cancer patients and is often associated with development of metastatic foci and resistant cell populations. Thus, recurrent breast cancer indicates a poor prognosis for the patient. Local tumor-derived 13762NF rat mammary adenocarcinoma cell clone MTF7(T20) was injected into the inguinal mammary fat pad and allowed to grow before surgical excision. Individual locally growing (primary) tumors were removed and established in short-term tissue culture. Corresponding local recurrences were excised after regrowth and established in short-term tissue culture. All sublines were tested for in vitro sensitivities to 5-fluoro-2'-deoxyuridine, Adriamycin, and ionizing X-irradiation. Using a clonogenic colony formation assay, responses of individual sublines ranged from 85 to 1500 ng/ml for Adriamycin and 65 to 10,000 nM for FdUrd. Some recurrences were significantly more resistant while others were more sensitive than the corresponding primary tumor lines. All recurrences had smaller 90% lethal dose values than the corresponding parent or primary tumor in response to Adriamycin; whereas, to 5-fluoro-2'-deoxyuridine, 90% lethal dose values revealed that most lines were quite resistant. Statistically significant differences in radiation survival were observed only for lines LR1a and LR5 (more sensitive). There was no apparent correlation between sensitivities to chemotherapy agents or X-irradiation and experimental metastatic potential in LR sublines. These dose-response data indicate that locally recurrent tumors are frequently, but not always, different from the original primary tumor in response to chemotherapy agents and ionizing X irradiation. Although an exact mechanism is unknown, it is likely that "selective" pressures which eliminate large numbers of cells, in this case surgery, change tumor composition so that recurrent tumors may no longer be equivalent to the tumor mass that was originally excised. This suggests that treatment strategies should be planned accordingly. PMID- 2969282 TI - Efficient transplantation of human non-T-leukemia cells into nude mice and induction of complete regression of the transplanted distinct tumors by ricin A chain conjugates of monoclonal antibodies SN5 and SN6. AB - In the present study, we established a dependable system by which human pre-B- and non-T/non-B-acute lymphoblastic leukemia (ALL) cells are efficiently transplanted into nude mice; the transplanted tumors provide a useful model for investigating the efficacy of antitumor agents in the in vivo therapy of human cancer. NALM-6 (a pre-B-ALL cell line) cells were transplanted under varying conditions as the pre-B-leukemia cells, whereas REH (a non-T/non-B-ALL cell line) cells were transplanted as the non-T/non-B-leukemia cells. Under optimal and near optimal conditions, 71 of 101 X-irradiated mice (70%) developed distinct tumors approximately 2 wk after i.d. inoculation of a mixture of NALM-6 cells and X irradiated human fibrosarcoma cells. Under the same conditions, 9 of 11 mice (82%) developed tumors following i.d. inoculation of REH cells admixed with X irradiated human fibrosarcoma cells. Examination of the tumor tissues demonstrated that the tumors are of leukemia origin but not of fibrosarcoma origin. To demonstrate the usefulness of the present tumors for investigating the efficacy of antitumor agents in the in vivo therapy of human cancer, immunotoxins were tested for their specific suppressive activity against growing tumors of the transplanted NALM-6 cells. To this end, monoclonal antibodies SN5 and SN6 which define a common ALL antigen, termed CALLA, and a novel leukemia-associated cell surface glycoprotein, termed gp160, respectively, were separately conjugated with the A-chain subunit of ricin, a plant toxin; CALLA and gp160 are expressed on the cell surface of various human non-T-leukemia cells including NALM-6 cells. The conjugates of SN5 and SN6 with ricin A-chain (RA) showed specific activity against the leukemia cells but not against control cells in an in vitro assay. To investigate their in vivo efficacy in suppressing tumor growth, nude mice which had been inoculated i.d. with NALM-6 cells 25 days in advance and bore distinct palpable tumors (5 to 6 mm in diameter) were divided into five groups. One group of mice was nontreated as a control. Each of the remaining four groups of mice was given an injection of one of the following agents: (a) purified control mouse IgG (IgG1); (b) purified antibodies SN5 (IgG1) and SN6 (IgG1); (c) control IgG-RA conjugate; or (d) SN5-RA and SN6-RA. Tumors in all mice of the first four groups including the untreated group grew continuously, causing the mice to die.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969283 TI - DR antigen expression on ovarian carcinoma cells does not correlate with their capacity to elicit an autologous proliferative response. AB - Expression of HLA-DR antigens by purified preparations of human ovarian carcinoma cells freshly isolated from surgical specimens was examined in parallel with the capacity of tumor cells to elicit a blastogenic response from autologous lymphocytes in mixed lymphocyte-tumor culture (MLTC) assay. Of 21 tumor preparations, 11 (52%) reacted with monoclonal antibodies 279 and/or 949 specific for a monomorphic determinant of HLA-DR antigens, with heterogeneous positivity, ranging between 30% and 95%. In this series of patients positive MLTC occurred in 8/21 individual experiments. The HLA-DR expression was proportionally similar in tumors giving positive MLTC (4/8 = 50%) and negative MLTC (7/13 = 53%). The lack of correlation between DR expression on tumor cells and stimulatory activity in autologous MLTC and the fact that DR-negative tumors could induce lymphocyte stimulation, support the hypothesis that blastogenesis occurs upon recognition of tumor-associated antigens, different from DR molecules, possibly tumor-specific antigens. PMID- 2969284 TI - Proliferative response of lymphocytes from ovarian cancer patients to autologous tumor cells. AB - Peripheral blood lymphocytes (PBL) from 43 patients with histologically confirmed ovarian carcinoma were stimulated in mixed lymphocyte-tumor culture (MLTC) with purified autologous tumor cells. Positive results, assessed as lymphocyte proliferation, were observed in 21 cases (48.8%). Lymphoid cells associated with ascitic fluid or infiltrating solid masses were in general less reactive than PBL as only 3/11 cases had positive MLTC. Tumor cells isolated from peritoneal effusions showed no significant difference in stimulatory potential as compared to the primary tumor. These results suggest that in an appreciable proportion of ovarian carcinoma patients (approximately 50%), lymphocytes have the potential to react to autologous tumor cells. Comprehension of the immunological mechanisms of antitumor resistance may have direct practical relevance for more effective treatment of neoplasms. PMID- 2969285 TI - Cellular immunological defects of chronic myelogenous leukaemics: partial dependence on busulphan therapy. AB - The PBMC from treated (n = 10) and untreated (n = 7) chronic phase CML patients were examined for their functional expression of helper cell-stimulating class II products, HLA-DR and -DP, and for their ability to induce suppression in normal PBMC. Although DR and DP were found to be functionally expressed in both groups of patients, a dysregulation of suppression induction was found in treated but not in untreated patients. Furthermore, the patients demonstrated a virtual absence of NK activity and severely depressed LAK activity which was equally striking in both treated and untreated patients and did not seem to be related to the presence of active suppression of cytotoxicity. Such defects in chronic phase CML patients may be relevant to the progression of their disease. Moreover, at least one of the cellular immunological defects, induction of suppressive cells, was not intrinsic to the disease, but appeared to be chemotherapy related. PMID- 2969287 TI - Coronary reperfusion with a new catheter in six patients with acute occlusion after angioplasty. AB - A new catheter was used in an attempt to re-establish coronary flow in six patients with acute occlusion after angioplasty and prior to emergency coronary artery bypass surgery. All patients suffered from severe chest pain, and the electrocardiogram showed signs of acute transmural ischemia. A reperfusion catheter manufactured by Advanced Cardiovascular Systems (ACS) has 36 side holes spirally arranged along the distal 10 cm. It is introduced over an exchange wire. After the catheter was positioned across the occlusion, five patients had complete relief of chest pain, and another patient had partial relief. The electrocardiogram normalized in all six patients (in one only initially). Two patients developed a limited myocardial infarction, but four did not. The time between introduction of the reperfusion catheter and aorta cross clamping varied between 1.5 and 6.5 hours, and there were no complications related to its use. We conclude that the ACS reperfusion catheter is useful in re-establishing coronary blood flow after failed angioplasty and prior to emergency coronary artery bypass surgery. PMID- 2969288 TI - Entrapment of an angioplasty balloon catheter: a case report. AB - This report presents a case in which an angioplasty balloon catheter became entrapped within the lumen of a coronary artery after rupture during percutaneous transluminal coronary angioplasty (PTCA). Prior to this report, balloon rupture had been considered a relatively benign occurrence. However, this case demonstrates that balloon rupture may lead to serious complications. PMID- 2969286 TI - Immunochemical characterization and ultrastructural localization of chondroitin sulfates and keratan sulfate in embryonic chick bone marrow. AB - Monoclonal antibodies directed against specific carbohydrate epitopes on chondroitin 4-/dermatan sulfate, chondroitin 6-sulfate, keratan sulfate, and a monoclonal antibody directed against the hyaluronate binding region were used to characterize proteoglycans extracted from embryonic chick bone marrow. About half of the proteoglycans separate into the high density fraction on a CsCl gradient. Glycosaminoglycan-specific antibodies recognize proteoglycans from all fractions; this includes an antibody directed against keratan sulfate. Some proteoglycans, principally in the high buoyant density fraction, contain sites recognized by the antibody specific for the hyaluronate binding region. Within limits of detection, all core proteins belong to the high-molecular-weight category, with weights in excess of 212 kD. Antibodies directed against chondroitin 4-/dermatan sulfate and against keratan sulfate primarily bind to extracellular matrix material located in the extracellular spaces and to matrix elements in the pericellular regions of fibroblastic stromal cells. The antibody that recognizes chondroitin 6-sulfate binds to sites on surfaces of fibroblastic stromal cells and also to extracellular matrix material. Little or no antibody binding is detected on surfaces of granulocytic cells. These studies indicate that chondroitin sulfate and keratan sulfate chains are both present in the proteoglycan extract. PMID- 2969289 TI - Saphenous vein graft rupture during percutaneous transluminal angioplasty. AB - Three cases (one fatal) of a rare angioplasty complication, vein graft rupture, are presented with a review of the literature. Caution is suggested when oversizing balloons for vein graft dilatation. PMID- 2969290 TI - Coronary angiography and angioplasty. PMID- 2969291 TI - Probe, a balloon wire: initial experience. AB - Coronary angioplasty is unsuccessful in less than 3-5% of cases because the balloon catheter fails to follow a guidewire that has traversed a lesion. Between June 1986 and August 1987, 31 lesions were unable to be crossed with at least two standard angioplasty catheters. Finally, a 2.0-mm-diameter Hartzler LPS (ACS) was utilized and successfully crossed and dilated 16 out of 31 lesions (52%). In the remaining 15 lesions, the Probe (USCI) 2.0 mm diameter x 1.5 cm long balloon wire was able to cross the lesions in 13 (82%) and successfully dilated 12. In one case, lesion rigidity prevented the balloon from expanding at 14 atm. A right coronary artery lesion was attempted in 11 cases, and a left anterior descending and circumflex artery lesion in two patients each. No complications were encountered. In seven out of 12 successful Probe cases, a larger balloon catheter was used to further dilate the artery. This new balloon wire has increased our success rate in severe stenoses and in tortuous vessels with severe distal lesions, in which presently available angioplasty equipment has failed. PMID- 2969292 TI - Evidence for specific association between class I major histocompatibility antigens and the CD8 molecules of human suppressor/cytotoxic cells. AB - Human T lymphocytes, metabolically labeled with 35S-cysteine and 35S-methionine, were reacted with the homobifunctional cross-linking reagent, dithiobis (succinimidyl propionate) (DSP). When detergent lysates from these cells were immunoprecipitated with a monoclonal antibody reactive with the CD8 antigen, a radiolabeled protein of approximately 44 kd was coprecipitated with the CD8 molecule. Immunoprecipitates from detergent lysates prepared without prior chemical cross-linking contained only the 33 kd CD8 molecule. Similar results were obtained when T lymphocytes or a cytotoxic T cell clone (T4T8Cl) were radiolabeled with 32P-orthophosphoric acid. The 44 kd CD8-associated protein was identified as the heavy chain of the class I major histocompatibility antigen by depletion in preclearing experiments with anti-class I MHC antibody and by peptide mapping. Further analyses indicated that the CD8-class I MHC association is due, in part at least, to disulfide bonding, which may be susceptible to cleavage during processing of cell lysates. PMID- 2969293 TI - Gastric mucosal lesions in streptozotocin-diabetic rats. AB - Formation of gastric mucosal lesions by streptozotocin-induced diabetes was investigated in rats. A single intravenous administration of streptozotocin in a dose of 65 mg/kg effectively produced hyperglycemia and damaged the gastric mucosa. Incidence and severity of mucosal lesions were progressively increased with time, from one to six weeks posttreatment. Microscopic lesions of the mucosa included hyperemia, desquamation of the surface epithelium with diffuse hemorrhage, and severe hemorrhage with localized erosion. Concurrent to the hyperglycemia, the histamine stimulated gastric H+-secretion was significantly decreased whereas pepsin secretion was not affected. Both soluble mucus and surface mucus gel were increased. The result suggests that the early lesion of gastric mucosa may be associated with the direct action of streptozotocin, the severity of which may be further aggravated by diabetic state. PMID- 2969294 TI - Monoclonal antibody DH12 reacts with a cell surface and a precursor form of the beta subunit of the human fibronectin receptor. AB - Monoclonal and polyclonal antibodies were raised against a placenta plasma membrane protein preparation, which was obtained by fractionation on Blue B dye matrix and by HPLC-anionexchange, and which was shown to contain fibronectin receptors. Immunochemical and functional evidence showed that monoclonal antibody DH12 recognized the beta subunit of the human fibronectin receptor on fibroblasts. This monoclonal antibody reacted with two proteins in Western blots and in double immune precipitations of whole cell preparations. Only the higher Mr protein became labeled by surface iodination of intact fibroblasts. The lower Mr protein is thought to be an intracellular precursor of the beta subunit of the fibronectin receptor. PMID- 2969295 TI - [Incidence of viral hepatitis in Czechoslovakia 1950-1985]. PMID- 2969296 TI - [Epidemiologic role of lactose-positive enterotoxigenic E. coli]. PMID- 2969297 TI - [The role of metronidazole in the treatment of anaerobic infections]. PMID- 2969298 TI - [Culture and identification of Gardnerella vaginalis]. PMID- 2969299 TI - [Comparison of the Staphytest and conventional methods in the diagnosis of staphylococci isolated from human urine]. PMID- 2969300 TI - [Helminthological study of children's sandboxes in Ceske Budejovice]. PMID- 2969301 TI - [Surgical treatment of laparoschisis and omphalocele]. PMID- 2969302 TI - Efficacy of teicoplanin as antimicrobial treatment of severe nosocomial infections caused by gram-positive bacteria: a preliminary study. AB - We used single daily intravenous teicoplanin as therapy for 12 severe nosocomial infections caused by gram-positive bacteria. A daily dosage of 3-6 mg/kg was usually adopted; however, in selected cases the dosage was increased to 8-9.5 mg/day on the basis of serum bactericidal monitoring. Most of these infections were life-threatening and included ventriculitis/meningitis (3 cases), sepsis (3 cases), mediastinitis (1 case) and extensive burn wound infection (1 case). Staphylococcus aureus was by far the most frequent pathogen and methicillin resistant strains were isolated in 7 out of 9 infections caused by this organism. The remaining isolates were Staphylococcus epidermidis, JK Corynebacterium, Streptococcus agalactiae and Propionilbacterium acnes. Additional antibiotics were used in 5 cases for concomitant gram-negative bacillus etiology (2 cases), granulocytopenia (2 cases), superinfection (1 case). Overall a clinical success and microbial eradication were documented in 100% and 91% of 12 cases, respectively. Except one case of fever, no other major adverse effect was observed and no patient required trial therapy discontinuation. In conclusion, our preliminary data seem to suggest a satisfactory activity of teicoplanin against nosocomial gram-positive infections. PMID- 2969303 TI - Seborrheic dermatitis in otherwise healthy patients and in patients with lymphadenopathy syndrome/AIDS-related complex: treatment with 1% bifonazole cream. AB - Recent studies reported in the literature have repeatedly pointed out the utility of imidazole derivatives, one of which is bifonazole, in the therapy of seborrheic dermatitis (SD). The cause of this pathological situation is still under discussion but the therapeutic success obtained with antimycotics speaks in favor of the pathogenetic importance of Malassezia ovale in this context. There is a high frequency of SD type complaints in patients with HIV infections in whom immune defenses against various pathogens, including yeasts, are deficient. We therefore verified the clinical efficacy of 1% bifonazole cream on 15 subjects with SD of the face, 9 of whom were affected with lymphadenopathy syndrome/AIDS related complex (LAS/ARC). The treatment was administered over a four-week period and gave good results in 12 patients. The best results, however, were observed in those suffering from LAS/ARC. PMID- 2969304 TI - Effect of uridine coadministration on 5'-deoxy-5-fluorouridine disposition in rats. AB - Uridine (UR) inhibits the metabolic activation of 5'-deoxy-5-fluorouridine (dFUR) to 5-fluorouracil (FU) by the intestinal pyrimidine nucleoside phosphorylases and could potentially reduce its intestinal toxicity. This study examined the effect of UR coadministration on the absorption and disposition of an oral dose of dFUR. Rats were given dFUR alone (500 mg kg-1) and dFUR (300 mg kg-1) plus UR (4.5 g kg 1) in a random crossover experiment. Simultaneous injection of a tracer dose of [6-3H]dFUR was used to asses the total body clearance (Cl) of dFUR. The absorption of UR was rapid and variable. The UR dose produced a maximal blood concentration of 80 micrograms/ml for UR and 100 micrograms/ml for its metabolite uracil (U). The absorption of dFUR was slower than UR, as indicated by its later time of maximal concentration. UR did not alter the Cl of dFUR, but reduced the absorption rate of dFUR from the gastrointestinal tract and significantly reduced the absolute oral bioavailability of dFUR from 55.2% to 33.4%. The effects of UR coadministration on the dFUR metabolite FU were opposite to those on dFUR; the FU availability was increased sixfold, and the elimination of FU was reduced. Based on the known competition between pyrimidine bases for their saturable metabolic enzymes, the increase in FU availability by UR coadministration was likely due to a competitive inhibition of FU metabolism by U. This study established the complex pharmacokinetic interactions between dFUR and UR and between their metabolites, which may be important in the modulation of dFUR activity by UR. PMID- 2969305 TI - Concentration and time dependence of the toxicity of fluorinated pyrimidines to HT 29 colorectal carcinoma cells. AB - To determine the optimal concentration time factors for the fluoropyrimidines 5 fluorouracil (FU), 5-fluorouridine (FUR), and 5-fluoro-2'-deoxyuridine (FUdR) in regional chemotherapy, we tested these drugs against the colorectal carcinoma cell line HT 29 at various dosages and exposure times. The measure of cytotoxicity used was the degree of inhibition of colony formation in soft agar after drug treatment compared with untreated control cells. Colonies were visible after 6 days of growth in soft agar, so the initial evaluation of toxicity was done at this time. Additional colonies were found 10 and 16 days after the first evaluation, so the dishes containing the treated cells were also evaluated for this delayed growth phenomenon ("regrowth"), which we considered to be due to a cell growth inhibition effect of the drugs rather than a cytocidal effect. Exposure times of the cells to the drugs ranged from 5 min to 24 h and the doses, between 0.01 and 1000 micrograms/ml. The toxicity of FUdR was concentration dependent, but its time dependence ceased after a relatively short exposure time. There was a cell population that was not susceptible to FUdR regardless of dose and exposure time; consequently, FUdR treatment was always accompanied by substantial regrowth of colonies. With FU and FUR, conditions could be achieved that resulted in complete cell death (no regrowth), but high concentrations and long exposure times were required with FU. With FUR, on the other hand, both cytostasis and cytotoxicity could be achieved with substantially lower doses and shorter exposure times than with FU. These results indicate that FUR has the potential to be an effective drug in chemotherapy protocols not involving systemic administration. PMID- 2969306 TI - Hemodynamic responses to atrial natriuretic factor in nephrectomized rabbits: attenuation of the circulatory consequences of acute volume expansion. AB - We investigated the hemodynamic responses to three doses of atrial natriuretic factor [human atrial natriuretic factor-(99-126)] (ANF) in nephrectomized rabbits anesthetized with ketamine and acepromazine. The influence of the different doses of the peptide on the hemodynamic consequences produced by acute volume expansion (0.9% NaCl, 1.4 ml/kg/min for 60 minutes) was also studied. All three dosages of ANF (0.001, 0.01, and 0.2 micrograms/kg/min for 20 minutes) significantly reduced blood pressure. With the lowest dose, the hypotensive effect was associated with reduction in systemic vascular resistance and no significant change in heart rate, stroke volume, central venous pressure, and hematocrit. In contrast, the intermediate and high doses, which resulted in markedly higher plasma levels, caused a significant decrease in heart rate, central venous pressure, and stroke volume; a slight rise in hematocrit; and no change in systemic vascular resistance. Volume expansion produced by saline infusion in an additional group of nephrectomized rabbits increased central venous pressure and decreased hematocrit. When ANF infusion was associated to volume expansion, each dosage of ANF was able to reduce the rise in central venous pressure, while only the higher dosage attenuated the progressive fall in hematocrit caused by volume expansion. Plasma volume, measured at the end of volume expansion was lower in the group treated with the highest dose of ANF than in the control animals (28.2 +/- 9 vs. 35.1 +/- 3 ml/kg, p less than 0.05). We conclude that 1) ANF induces significant hemodynamic effects independently from its renal action.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969307 TI - Changes in myofibrillar content and Mg-ATPase activity in ventricular tissues from patients with heart failure caused by coronary artery disease, cardiomyopathy, or mitral valve insufficiency. AB - Force development and shortening by cardiac muscle occur as a result of the interaction between actin and myosin within the myofibrillar lattice. This interaction is dependent upon intracellular ionized calcium and is controlled by the troponin-tropomyosin regulatory proteins situated along the actin filament. In this study, we compared the myofibrillar content and myofibrillar Mg-ATPase activity of normal human ventricular muscle with that of ventricular muscle from patients in end-stage failure caused by coronary artery disease or cardiomyopathy and ventricular muscle from patients with heart failure due to mitral valve insufficiency. The results show that the amount of myofibrillar protein (mg/g wet wt ventricle) in hearts in end-stage failure (coronary artery disease and cardiomyopathy) is significantly lower compared with normal hearts and hearts in failure due to mitral valve insufficiency. However, the Mg-ATPase activity of myofibrils from hearts in both end-stage failure and failure due to mitral valve insufficiency is significantly lower compared with myofibrils from normal hearts. The data suggest that the reduction in the amount of myofibrillar protein in ventricular tissue is a pivotal event that may be responsible for the progression of heart disease to the point of end-stage failure. PMID- 2969308 TI - Increase in cross-linking of type I and type III collagens associated with volume overload hypertrophy. AB - Types I, III, IV, and V collagen were isolated and characterized from eight normal dog hearts and seven with volume-overload hypertrophy. Animals with volume overload hypertrophy were killed at a time when left ventricular end-diastolic pressure and stiffness were increased. The collagens were characterized by solubility properties, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, and enzyme-linked immunosorbent assay. The percentage of collagen obtained from canine left ventricles was decreased from 32.4% in normal hearts to 15.0% in hypertrophied hearts. We attribute this to a diminution in the extractability of types I and III collagen, which fell from 199.5 mg type I/g collagen and 76.4 mg type III/g collagen in normal hearts to 83.5 mg type I/g collagen and 26.4 mg type III/g collagen in hypertrophied hearts. The amount of types IV and V collagen isolated remained constant in both the control and arteriovenous shunt hearts averaging 15.7 mg type IV/g collagen and 32.1 mg type V/g collagen in control hearts and 12.5 mg type IV/g collagen and 28.9 mg type V/g collagen in hypertrophied hearts. The reduction in quantity of types I and III collagen probably reflects a greater degree of cross-linking in these two types of collagen. Cyanogen bromide peptide analysis confirmed that there was an increase of high molecular weight cross-linked peptides from 3.96% in normal samples to 8.88% in hypertrophied samples. We conclude that cross-linking of types I and III collagen increases in volume-overload hypertrophy and that this is associated with a rise in diastolic stiffness. PMID- 2969309 TI - Microvascular effects of atrial natriuretic factor: interaction with alpha 1- and alpha 2-adrenoceptors. AB - The cremaster skeletal muscle of anesthetized rats was denervated and extended with intact circulation into a tissue bath. Intravital microscopy was used to measure microvessel diameter at three different anatomical levels within the microcirculation: large distributing arterioles (x control diameter = 100 +/- 7 micron), large capacitance venules (147 +/- 8 micron), and small terminal arterioles (17 +/- 1 micron). Norepinephrine (NE) was added to the cremaster bath to produce intermediate reductions in diameter of large arterioles and venules (55% and 38% of maximum constriction, respectively). In the presence of NE tone, bath-added atrial natriuretic factor (ANF) produced concentration-dependent dilation of both arterioles and venules. Arteriolar IC25 = 18 pmol and IC50 = 1.2 X 10(-10) M; venules exhibited similar sensitivity. However, the highest ANF concentration examined (10(-7) M) only reversed NE-induced tone by 70%. In a second large vessel group ANF completely reversed constriction induced by the alpha 1-adrenoceptor agonist, phenylephrine, in the presence of 5 X 10(-7) M yohimbine. However, vessels constricted with the alpha 2-receptor agonist UK 14,304 (in the presence of 10(-8) M prazosin) were insensitive to ANF. A third group of terminal arterioles, which possess considerable spontaneous "intrinsic" tone, were studied in the absence of alpha-receptor agonists. Significant dilation occurred at greater than 10(-7) M, and the maximal response was only 25% of complete dilation with adenosine. These data indicate that ANF exhibits a high potency and selectivity for reversal of alpha 1-adrenoceptor-mediated constriction of large arterioles and venules. Constriction produced by alpha 2 adrenoceptor occupation or by nonadrenergic "intrinsic" mechanisms appears to be insensitive to ANF. We propose that the ability of ANF to reduce microvascular resistance depends on the relative contribution of alpha 1-, alpha 2-, and intrinsic vasoconstrictor components to the prevailing level of smooth muscle tone. Differences in these components among regional circulations and between arterial and venous smooth muscle may contribute to the systemic hemodynamic pattern produced by ANF. PMID- 2969310 TI - Intestinal microvascular growth during maturation in diabetic juvenile rats. AB - To determine if intestinal microvascular growth is impaired in diabetic juvenile animals, a segment of the terminal ileum was marked and the microvasculature of this segment observed at the age of 5 weeks and again at the age of 10-11 weeks in normal and diabetic Sprague Dawley rats. Diabetes was induced by streptozotocin after the first observation period and the plasma glucose concentration exceeded 500 mg% by the age of 10-11 weeks. Microvascular growth was quantitated by measurements of the number, length, and maximally dilated inner diameters of specific arterioles and by intercapillary distances in the marked intestinal region at both ages. Although intestinal enlargement was much greater in diabetics, there was no change in the number of arterioles during maturation and intercapillary distances were equivalent in diabetic and normal rats. In normal and diabetic animals, the arteriolar length increased to match bowel elongation, however, increases in bowel and arteriolar lengths in diabetic animals were about twice that of normal rats. During juvenile maturation, the maximally dilated inner diameters of the small arterioles in diabetic animals were increased compared with their normal counterparts. Thus, arteriolar growth during maturation is characterized by changes in the length but not in the number of vessels in intestine of both normal and diabetic rats. The perfusion of about 90% more tissue by mass for each arteriole in diabetic rats is facilitated by arteriolar dilation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969312 TI - Guidelines for percutaneous transluminal coronary angioplasty. A report of the American College of Cardiology/American Heart Association Task Force on Assessment of Diagnostic and Therapeutic Cardiovascular Procedures (Subcommittee on Percutaneous Transluminal Coronary Angioplasty). PMID- 2969311 TI - Experimental validation of Doppler echocardiographic measurement of volume flow through the stenotic aortic valve. AB - In aortic stenosis, evaluation of aortic valve area by the continuity equation assumes that the volume of flow through the stenotic valve can be measured accurately in the left ventricular outflow tract. To test the accuracy of Doppler volume-flow measurement proximal to a stenotic valve, we developed an open-chest canine model in which the native leaflets were sutured together to create variable degrees of acute aortic stenosis. Left ventricular and aortic pressures were measured with micromanometer-tipped catheters. Volume flow was controlled and varied by directing systemic venous return through a calibrated roller pump and back to the right atrium. Because transaortic volume flow will not equal roller pump output when there is coexisting aortic insufficiency (present in 67% of studies), transaortic flow was measured by electromagnetic flowmeter with the flow probe placed around the proximal descending thoracic aorta, just beyond the ligated arch vessels. In 12 adult, mongrel dogs (mean weight, 25 kg), the mean transaortic pressure gradient ranged from 2 to 74 mm Hg, and transaortic volume flow ranged from 0.9 to 3.2 l/min. In four dogs, electromagnetic flow that was measured distal to the valve was accurate compared with volume flow determined by timed collection of total aortic flow into a graduated cylinder (n = 24, r = 0.97, electromagnetic flow = 0.87 Direct +0.13 l/min). In eight subsequent dogs, electromagnetic flow was compared with transaortic cardiac output measured by Doppler echocardiography in the left ventricular outflow tract as circular cross sectional area [pi(D/2)2] x left ventricular outflow tract velocity-time integral x heart rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969313 TI - Clinical effectiveness of dermatomal evoked cerebrally recorded somatosensory responses. AB - Among 129 patients with spine pain and radiculopathy in both the cervical and lumbar region, there were significant differences between the upper and lower extremity DSER groups, noting the higher number of normal studies by our criteria in the upper extremities as compared to the lower extremities. This difference may be due to the intertwining of the "nerve circuitry" of the input pathways. It appears that the dermatomal somatosensory evoked response is a study of low sensitivity and of high specificity. PMID- 2969314 TI - The plasma proteases, thrombin and plasmin, degrade the proteoglycan of rabbit aorta segments in vitro: an integrated ultrastructural and biochemical study. AB - The effects of plasmin and thrombin on the proteoglycan component of uninjured and deendothelialized rabbit thoracic aorta were examined in vitro. Aortas labelled with 35S in vivo, were exposed to each protease and the 35S-products released from the vessel were analysed. Aortas exposed to the same enzymes were examined by transmission electron microscopy of ruthenium red-stained sections, and the proteoglycan content evaluated by morphometric analysis. In uninjured vessels with endothelium intact, there was no evidence of release of proteoglycan by either enzyme unless very high concentrations were used. High concentrations of both plasmin and thrombin induced vacuolation of endothelial cells, and changes in the fine structure of smooth muscle cells. In contrast exposure of de endothelialized vessels to smaller concentrations of plasmin or thrombin resulted in a significant loss of proteoglycan, and less marked changes in the morphology of the smooth muscle cells. Thrombin treatment released 35S-labelled products containing chondroitin-, dermatan- and heparan-sulphates whereas plasmin released products containing only chondroitin- and dermatan-sulphates together with other, unknown 35S-labelled products. These observations indicate the potential of non specific, plasma-derived proteases to degrade arterial connective tissue. PMID- 2969315 TI - The effects of the long-acting angiotensin-converting enzyme inhibitor cilazapril on casual, exercise, and ambulatory blood pressure. AB - We assessed blood pressure (BP) and heart rate (HR) responses in a double-blind, randomized study comparing cilazapril, a long-acting, nonsulfhydryl-group converting enzyme inhibitor, with placebo in 18 patients with mild to moderate (sitting diastolic BP, 95 to 114 mm Hg) essential hypertension. The BP and HR parameters were evaluated at rest (casual, 24 hours after administration), during treadmill exercise testing (Bruce protocol), and with 24-hour noninvasive ambulatory BP monitoring. These assessments were made after a 4-week drug washout period and after 8 to 12 weeks of therapy. After 8 weeks of therapy with cilazapril (mean dose 3.6 +/- 0.9 mg/day), casual BP decreased 19/11 mm Hg (p less than 0.01), whereas placebo lowered BP by 4/5 mm Hg (difference not significant) compared with the baseline period. The casual HR was modestly (7 beats/min) but significantly (p less than 0.05) lowered by cilazapril monotherapy. Exercise BP was reduced by cilazapril (reduction at peak HR, 23/11 +/- 10/5 mm Hg; p less than 0.05), and exercise HR was unchanged. Compared with baseline, the duration of exercise was improved with cilazapril but not with placebo (1.0 minute vs -0.2 minute; p less than 0.05). Twenty-four-hour mean, awake, and sleep BPs were reduced with cilazapril with the most impressive reduction occurring during the awake period (19/12 mm Hg; p less than 0.01). These data demonstrate that cilazapril lowers casual, exercise, and ambulatory BP with a modest but significant improvement in exercise time. Thus cilazapril may be particularly effective in the physically active hypertensive patient. PMID- 2969316 TI - Decline of atrial natriuretic peptide release in dogs during sustained rapid cardiac pacing. AB - 1. To assess the ability of the atria to maintain elevated plasma concentrations of atrial natriuretic peptide (ANP), the temporal changes in plasma ANP concentrations were studied in seven chloralose-anaesthetized dogs during 4 h of sustained rapid cardiac pacing. 2. Heart rate increased from 124 +/- 26 (mean +/- SEM) to 278 +/- 28 beats/min for the 4 h duration of rapid cardiac pacing. Mean pulmonary wedge pressure increased from 3.6 +/- 1.8 to 17.4 +/- 7.1 mmHg at 30 min (P less than 0.01) and mean right atrial pressure rose from -1.7 +/- 1.9 to 2.0 +/- 2.8 mmHg at 30 min (P less than 0.01). Both remained constant at these elevated pressures for the entire 240 min of rapid pacing. 3. Arterial ANP concentrations increased in all dogs from 87 +/- 11 to a maximum of 1263 +/- 592 pmol/l at 30 min (P less than 0.01), falling to 411 +/- 42 pmol/l after 60 min and to 146 +/- 70 pmol/l after 240 min of rapid continuous pacing (P less than 0.01 compared with 30 min). Coronary sinus ANP concentrations showed a similar pattern, rising from 241 +/- 79 to a maximum of 1837 +/- 203 pmol/l after 30 min (P less than 0.01). These peak values likewise were not sustained, falling to 962 +/- 198 pmol/l after 60 min and 297 +/- 41 pmol/l after 240 min of rapid pacing (P less than 0.01 compared with 30 min). 4. It is concluded that atria are unable to maintain the peak concentrations of ANP reached after 30 min of rapid pacing despite persistently elevated atrial pressures. PMID- 2969317 TI - Atrial natriuretic peptide released by rapid ventricular pacing in dogs does not cause a natriuresis. AB - 1. The relationships between the haemodynamic, renal and endocrine changes induced by rapid ventricular pacing were studied in ten chloralose-anaesthetized dogs paced from the right ventricular apex for 60 min at 250 beats/min. 2. Pacing increased mean right atrial and mean pulmonary wedge pressure (P less than 0.05), and decreased cardiac output and mean arterial pressure (P less than 0.05). 3. Coronary sinus atrial natriuretic peptide (ANP) concentrations were approximately fourfold greater than arterial concentrations; both increased markedly during pacing (P less than 0.01). Plasma concentrations of arginine vasopressin and plasma renin activity did not change significantly. 4. Urine flow and free water clearance increased during the latter 30 min of pacing (P less than 0.05). There was no significant change in sodium clearance despite high sustained concentrations of ANP. 5. Without the availability of specific inhibitors of ANP release or action, we are unable to exclude the possibility that ANP may have prevented sodium clearance from otherwise decreasing during rapid ventricular pacing. Nevertheless, the dissociation between elevated ANP concentrations and natriuresis in this study indicates that a rise in ANP concentrations per se is not sufficient to produce a natriuresis. PMID- 2969318 TI - Atrial natriuretic peptide and catecholamines in peripheral blood as indicators of cardiac dysfunction in the general population. AB - 1. From a screened cohort of 644 67-year-old men, drawn from the population of Gothenburg, 42 men with presumed cardiac dyspnoea were selected and compared with 45 random controls. 2. Cardiac function was evaluated by echocardiography and other non-invasive methods, and the potential of peripheral venous concentrations of immunoreactive atrial natriuretic peptide (IrANP), noradrenaline and adrenaline in revealing cardiac dysfunction was investigated. 3. Concentrations of venous IrANP and noradrenaline were both significantly, but weakly, related to the degree of dyspnoea and to pulmonary congestion. 4. IrANP, but not catecholamines, was also related to other indices of heart failure. The correlations were, however, due solely to increased immunoreactivity in men with severe dyspnoea; in most subjects with mild to moderate dyspnoea the level of IrANP was similar to that of the controls. 5. It is concluded that, in a population sample, the peripheral venous level of IrANP is more closely related to cardiac function than are catecholamines. Yet, the level of IrANP is not a very sensitive marker of cardiac dysfunction, suggesting that stimulation of alpha-human atrial natriuretic peptide release is a late phenomenon in the development of cardiac failure. PMID- 2969319 TI - Atrial natriuretic peptide and total exchangeable body sodium: relationships in rats with chronic myocardial infarction. AB - 1. The relationship between plasma atrial natriuretic peptide (ANP) and body sodium was determined in rats 1 month after myocardial infarction induced by coronary artery ligation. After operation rats received a normal or a low salt diet, and total exchangeable body sodium was measured sequentially. 2. Rats with infarction receiving a normal salt intake did not retain sodium when compared with sham-operated controls. Rats receiving a low salt diet had a 10% decrease in body sodium (P less than 0.01). The decrease was the same in rats with infarction as in controls. 3. Plasma ANP was similar in control rats irrespective of salt status. Plasma ANP levels were markedly elevated in rats with infarction irrespective of salt status (P less than 0.01). 4. The rise in plasma ANP was correlated with cardiac hypertrophy and infarct size in animals fed both normal and low salt diets. However, there was no relationship between plasma ANP and exchangeable body sodium. 5. These results suggest that in this model of heart failure plasma ANP is raised by increased left atrial stretch in proportion to the severity of left ventricular dysfunction. In contrast, plasma ANP concentrations do not appear to be elevated as a consequence of increased right atrial pressure caused by sodium retention and expanded extracellular volume. PMID- 2969320 TI - Changes in lipoprotein composition in women receiving two low-dose oral contraceptives containing ethinylestradiol and gonane progestins. AB - In a prospective double-blind study of the effects of two low-dose oral contraceptives (OCs) on lipid and lipoprotein metabolism, two groups of eighteen young healthy women selected at random were submitted to a six months' use of either monophasic ethinylestradiol (EE) + desogestrel (DG) or triphasic EE + levonorgestrel (LNG). Total cholesterol (C), triglycerides (TG), phospholipids (PL), low density lipoprotein-cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) together with apoproteins A-I, A-II, B and lecithin cholesterol acyltransferase activity (LCAT) were determined in serial plasma samples collected before, at three and six months during, and one month after OC use. Cholesterol and apoproteins (A-I, A-II, B) composition of lipoproteins (HDL 2, HDL-3, LDL) isolated by ultracentrifugation were additionally determined. On Mono-EE + DG, plasma TG (+39.3%, +45.6%), PL (+21.9%, +16.8%) and apo A-I (+35.5%, +23.3%) levels were significantly increased at 3 and 6 months of use; plasma HDL-C (+24%) and Apo A-II (+21.4%) were transiently increased at 3 months. Lipid and apoprotein composition of HDL-2, HDL-3 and LDL were unchanged. On Tri EE + LNG, a slight but not significant decrease in HDL-C was observed throughout the study while other plasma lipids and apoproteins were unchanged. Ultracentrifugation revealed a lower content of C (-44.2%) and apo A-I (-44.6%) in HDL-2. LCAT activity expressed as molar esterification rate (MER) rose in a more sustained way during EE + DG use than during EE + LNG treatment. Covariance analysis shows a further significant difference between results of both treated groups for the Apo AI/Apo B ratio that was increased by Mono-EE + DG. PMID- 2969321 TI - Long-term effects of MPA on human progeny: intellectual development. AB - Tests of verbal and spatial ability were done on 450 boys and 537 girls in their late teens of whom 73 and 97, respectively, had been exposed in utero to MPA. Exposed boys achieved higher raw scores than controls on verbal and spatial tests but the differences were explained by their more favorable demographic and social characteristics. Exposed girls did not differ from controls. Although, mothers of exposed boys reported that their offspring talked and walked later than controls, our results support the hypothesis that intrauterine exposure to MPA at contraceptive doses has no long-term effect on intellectual development. PMID- 2969322 TI - [Renal artery angioplasty in the treatment of renovascular hyper- tension]. PMID- 2969323 TI - Defective catabolism of D-glucose and L-glutamine in mouse pancreatic islets maintained in culture after streptozotocin exposure. AB - We recently described a preferential reduction of the secretory response to nutrient secretagogues (glucose; leucine plus glutamine) in islets maintained in culture after in vitro exposure to streptozotocin (SZ). The present study is an attempt to further clarify the biochemical mechanisms behind this defective insulin response. Mouse pancreatic islets were collagenase isolated and, after 4 5 days in culture, exposed during 30 min at 37 C to 1.8 mM SZ or vehicle alone (controls). The islets were subsequently cultured for 7 days in medium RPMI 1640 plus 10% calf serum, before the enzymatic and metabolic studies were performed. The activities of the glycolytic enzymes, hexokinase, glucokinase, and glyceraldehyde 3-phosphate dehydrogenase, were similar in the control and SZ exposed islets. The relative amount of cytosolic and mitochondria-bound hexokinase was also unaffected by SZ. However, there was a 30-40% decrease in the activity of NAD+- and NADP+-dependent glutamate dehydrogenase and glutamate aspartate transaminase in the SZ-treated islets. This coincided with a 40% decrease in L-[U-14C]glutamine oxidation in the SZ-treated islets. The D-glucose catabolism was further examined in the presence of D-[5-3H] and D-[6-14C] glucose. There was no difference between control and SZ islets in terms of glucose utilization at either 1.7 or 16.7 mM glucose. The oxidation of D-[6 14C]glucose was nevertheless decreased by more than 50% in SZ islets incubated at 16.7 mM (but not 1.7 mM) glucose. Altogether, these converging observations suggest a perturbation of distal regulatory processes, apparently at the mitochondrial level, in the D-glucose and L-glutamine catabolism of SZ-exposed islets. Whether this reflects a primary action of SZ on the islet mitochondria, or an inhibitory effect of SZ on the synthesis of mitochondrial enzymes, as a result of nuclear DNA damage, remains to be elucidated. PMID- 2969324 TI - Structural and immunohistochemical characterization of insulin-like growth factor I and II receptors in the murine central nervous system. AB - The description of the cellular localization of insulin-like growth factor (IGF) receptors in the central nervous system (CNS) remains incomplete, as do the descriptions of changes in their characteristics with respect to different developmental stages. We, therefore, performed affinity labeling studies in microsomal membrane preparations of adult and fetal rat brain and liver tissues with [125I]IGF-I and [125I]IGF-II. These studies demonstrated tissue- and developmental stage-specific structural variants of type I receptor alpha subunits as well as type II receptors. The adult rat brain type I alpha-subunit had an apparent mol wt (Mr) of 127,000, whereas those of adult and fetal rat liver measured 140,000. Fetal rat brain microsomes, however, had two types of type I receptor alpha-subunits measuring 130,000 and 120,000 Mr. The larger subunit from fetal brain consistently migrated at an apparent Mr of 3,000, greater than subunits from adult brain. Both type I and II receptors were more abundant in fetal liver and brain than in adult tissues. Affinity labeling was also performed directly to monolayers of cultured fetal brain neurons and newborn astrocytes. These studies detected both type I and II receptors on the surfaces of both types of cells. However, only the high Mr (140,000) form of the type I alpha-subunit was detected in cultured CNS cells, suggesting that expression of low Mr variant receptors is altered in vitro. Type II receptors were demonstrated by immunohistochemistry in adult rat hypothalamic neurons. However, the majority of neurons did not react with type II receptor antibody. This finding implies that only a minority of hypothalamic neurons are capable of responding to IGF-II via type II receptors. On the other hand, all astrocytes had striking type II receptor immunoreactivity. This signifies a more general biological role for this receptor in astrocytes compared with neurons. These results suggest that different tissue-, developmental stage-, and cell-specific processes are mediated by IGF receptors and suggests new directions in which to explore potential biological actions for these receptor-ligand systems in the CNS. PMID- 2969325 TI - Insulin-like growth factor I and insulin potentiate luteinizing hormone-induced androgen synthesis by rat ovarian thecal-interstitial cells. AB - We tested the hypothesis that insulin-like growth factor I (IGF-I) and insulin play a role in androgen production by rat ovarian thecal-interstitial cells. Collagenase/DNase-dispersed rat ovarian thecal-interstitial cells obtained from immature hypophysectomized Sprague-Dawley rats were cultured at a concentration of 10(6) cells/ml in serum-free medium in the presence of increasing concentrations of LH, IGF-I, or insulin. The medium was replaced every 48 h, and the androsterone concentration in the culture supernatants was used as an index of androgen production. In the absence of added hormones (control) androsterone levels were consistently less than 0.1 ng/ml. Increasing concentrations of LH stimulated androsterone synthesis in a dose-dependent manner. IGF-I, in the absence of LH, did not significantly increase androsterone levels above control values. However, when combined with 10 ng/ml LH, IGF-I increased androsterone synthesis above levels seen with LH alone in a dose-related fashion: for example, the peak androsterone levels seen with LH and 100 ng/ml (13 nM) IGF-I at 96 h of culture were significantly greater than the peak level seen with 10 ng/ml LH alone (302 +/- 71 vs. 17 +/- 7 ng/ml; P less than 0.0125). Similarly, while insulin alone did not increase androsterone synthesis above control values, androsterone concentrations were increased by insulin in combination with 10 ng/ml LH; a peak value of 240 +/- 67.7 ng/ml was observed at 96 h of culture with 100 ng/ml (18 mM) insulin (P less than 0.025 vs. LH alone) Androsterone levels were slightly less with insulin than with IGF-I, but this difference was not significant. The combination of IGF-I and insulin did not increase levels of androsterone synthesis above those observed with each hormone alone. IGF-I bound to a high affinity binding site on ovarian cell monolayer cultures with an apparent binding affinity of 1.3 x 10(-9) M. Insulin also competed for binding with radiolabeled IGF-I in a dose-dependent manner, but the affinity of insulin was approximately 500-fold less; half-maximal inhibition of [125I] IGF-I binding occurred with an insulin concentration of approximately 300 nM (or approximately 1700 ng/ml). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of thecal interstitial cell monolayers affinity labeled with radiolabeled IGF-I in the absence and presence of unlabeled hormone revealed proteins with characteristics of type I IGF receptors. Affinity labeling to a protein of a relative molecular mass of approximately 45,000 was also noted, probably representing IGF carrier proteins synthesized by thecal-interstitial cell monolayers.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969326 TI - Specific, high affinity receptors for insulin-like growth factor II in the rat kidney glomerulus. AB - Rat renal glomeruli were isolated by a technique involving kidney perfusion with a solution containing magnetic iron oxide particles, followed by homogenization, sieving, and concentration over a strong magnet. Isolated glomeruli were treated with 1% Triton X-100 to solubilize plasma membrane components, while insoluble basement membrane components were removed by centrifugation. [125I]Insulin-like growth factor II (IGF-II) binding to this preparation was competitively inhibited by increasing amounts of unlabeled IGF-II, with 50% inhibition at an IGF-II concentration of 1 ng/ml. [125I]IGF-II was covalently cross-linked with disuccinimidyl suberate to its receptor in rat renal glomeruli and a specific high mol wt (255,000) band could be identified on autoradiograms of dodecyl sulfate-polyacrylamide gels. [125I]IGF-II binding and cross-linking to this band was inhibited by a polyclonal antibody against the type II IGF receptor. These results demonstrate for the first time that the isolated rat renal glomerulus contains a high affinity receptor for IGF-II. PMID- 2969327 TI - Hepatic vasopressin receptor: differential effects of divalent cations, guanine nucleotides, and N-ethylmaleimide on agonist and antagonist interactions with the V1 subtype receptor. AB - Previously, we reported that magnesium (Mg2+) enhanced the binding affinity of arginine vasopressin [( 3H]AVP) to a single class of sites in rat liver microsomes. In the present study we have examined the effects of divalent cations and guanine nucleotides on the binding characteristics of both the nonselective agonist and the V1 receptor-selective antagonist, d(CH2)5Tyr(Me)-[3H]AVP, to microsomal and plasma membrane fractions of rat liver. At a subsaturating concentration (100 pM) of [3H]AVP, divalent cations increased specific binding in a concentration-dependent manner with the following rank order of potency: Co2+ greater than Mn2+ greater than Ni2+ greater than Mg2+ greater than Ca2+ = control. The maximal effect for Mg2+ was evident at 1 mM, a physiologically relevant concentration. In contrast, binding of the V1 receptor antagonist (at a subsaturating concentration of 10 pM) was inhibited by divalent cations, the rank order of potency being Mn2+ greater than Co2+ greater than Ca2+ greater than Mg2+ greater than Ni2+. The inhibitory effects of divalent cations were of lesser magnitude (up to 60%) compared to the stimulation of agonist binding (up to 700%). Mg2+ enhanced the affinity of [3H]AVP (Kd was decreased from approximately 2 nM to 133 pM), while the affinity of the [3H]V1 antagonist was decreased (Kd was increased from 10 to 95 pM). Scatchard analysis of saturation data (Mg2+ present) revealed similar maximum binding values for the binding of radiolabeled agonist and antagonist, indicating that AVP receptors in rat liver are mostly of the V1 subtype. Competition experiments between V1/V2-specific AVP analogs with either the radiolabeled agonist or antagonist also indicated the presence of predominantly V1 receptor sites in rat liver microsomes. The properties of plasma membrane receptor sites were similar to those of the microsomal sites, except that the density of receptors was higher in the former. In both equilibrium and competitive inhibition experiments GTPase-resistant analogs of guanine nucleotides, GTP gamma S and GDP beta S, decreased the affinity of the agonist for the receptor, but not that of the antagonist. Treatment of membranes with 0.2 mM N-ethylmaleimide (NEM) reduced the maximum binding of [3H]AVP and abolished the GTP gamma S-evoked decrease in agonist-binding affinity. In contrast, antagonist binding was unaffected by NEM. NEM pretreatment failed to influence the divalent cation-dependent increase in agonist-binding affinity. The results provide direct evidence for the existence of a high and a low affinity state of the hepatic V1 receptor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969328 TI - A reversed-phase high-performance liquid chromatographic method for the simultaneous determination of serum concentrations of cyproterone acetate and 15 beta-hydroxycyproterone acetate. AB - A reversed-phase high-performance liquid chromatographic method for the simultaneous determination of cyproterone acetate (CPA), 15 beta hydroxycyproterone acetate (15 beta-OH-CPA) and cyproterone (CP) was reported. This method was specific, sensitive, precise, easy and rapid for determination of the serum concentrations of these steroids in patients receiving CPA. Although no peak corresponding to CP was observed for serum, peaks corresponding to CPA and 15 beta-OH-CPA were detected and well separated in all subjects undergoing long term CPA therapy. In these patients, there seemed to be a dose-dependent relationship between the amount of CPA administered and the serum concentrations of these steroids, and the serum concentrations of CPA were either similar or low compared with those of 15 beta-OH-CPA. In conclusion, this simplified method is thought to be very valuable for studies on the pharmacokinetics of CPA and 15 beta-OH-CPA, and on the relationship between the CPA dosage and the therapeutic or side effects on adrenal and gonadal steroid production. PMID- 2969329 TI - Bromocriptine treatment in anovulation with decreased ratio of follicle stimulating hormone to luteinizing hormone and with hyperandrogenism. AB - Twenty-six anovulatory women of polycystic ovarian (PCO)-type were treated with bromocriptine (Br) at a daily dose of 5 mg for 2 months. Ovulatory cycles were resumed in 18 (69.2%) women (Br-responders). No difference between pretreatment serum levels of FSH, LH, PRL and estradiol and FSH/LH ratios in Br-responders and nonresponders was observed. The geometric mean of circulating androstenedione (A dione) in Br-responders (2.58 ng/ml) appeared higher than that in nonresponders (2.11 ng/ml) but was not statistically significant. The geometric mean of dehydroepiandtrosterone sulfate (DHEA-S) in Br-responders (1652 ng/ml) was lower (p less than 0.01) than that in nonresponders (2582 ng/ml). The ratio of DHEA-S to A-dione (D/A ratio) exhibited a highly significant between-group difference (p less than 0.001) (646 and 1222 for Br-responders and nonresponders, respectively). Br-nonresponders with high DHEA-S levels and D/A ratios tended to hyperproduction of adrenal androgen, and Br-responders with high A-dione levels and low D/A ratios to hyperproduction of ovarian androgen. The present study indicates that Br is effective in PCO-type women presumably with ovarian androgen hyperproduction. The efficacy of Br, when applied to PCO-type women, could be predicted with their D/A ratios. PMID- 2969330 TI - Study of beta-glucosidase from Helix pomatia by active site-directed inhibitors. AB - This work describes the purification of a beta-glucosidase (beta-D-glucoside glucohydrolase EC 3.2.1.21) from the digestive juice of Helix pomatia and the study of the enzyme's active site by using different reversible and irreversible inhibitors. The catalytic constants of arylglycosides and their pH-dependent variations have also been determined. The inhibition studies demonstrate that conduritol epoxides are irreversible inhibitors of beta-glucosidase from the digestive juice of H. pomatia, and that nojirimicin shows tight binding with glucosidase: the formation and dissociation of the enzyme-inhibitor complex (dissociation constant 1.1 mumol/1) required several minutes. PMID- 2969331 TI - A study of glucose handling by Buddhist monks. AB - Fourteen Buddhist monks and comparable male subjects were studied in relation to their handling of glucose after a meal (consisting of 1190 kcal, 29 g protein, 21 g fat and 221 g carbohydrate) and afterwards subjected to an oral glucose tolerance test (oGTT). The time course of blood glucose levels after the meal indicated that the monks had enhanced absorption and utilization of glucose. The monks were also found to have increased tolerance to glucose on oGTT. In addition the mean total serum cholesterol level in the monks (157.2 +/- 5.53 mg/dl) was found to be significantly higher than that of the control subjects (117.4 +/- 2.85 mg/dl). PMID- 2969332 TI - Variable region genes in the human T-cell rearranging gamma (TRG) locus: V-J junction and homology with the mouse genes. AB - The locus of the human T cell rearranging gamma (TRG) or T cell receptor gamma chain genes comprises at least 14 variable genes (TRGV) belonging to four subgroups, five joining segments (TRGJ) and two constant region genes (TRGC). Nine V gamma genes belong to subgroup I, whereas subgroups II, III and IV each consists of a single gene respectively designated V9, V10 and V11. T cells expressing the gamma chain (TRG+) and recognized by the anti-Ti gamma A monoclonal antibody have been shown to rearrange the V9 gene. In order to assess the N diversity at the V-J junction in the TRG+ cells, the germline sequences of the segments involved in the V-J rearrangements must be known. In this paper, we report the sequences of the germline V9 and V10 genes. Comparison of the V-J junction and N region from transcripts or rearranged TRG genes belonging to the different subgroups shows no evidence of D segments in the human TRG locus. Sequences of the rearranged V11 gene from the JM cell line and those of the VA and VB pseudogenes, located upstream of V9 and V11 respectively, are given. Our results bring the number of human V gamma genes whose sequence is known to 13 and reveal unexpected homology with the mouse V gamma genes. PMID- 2969334 TI - Responses of plasma human atrial natriuretic factor to high intensity submaximal exercise in the heat. AB - No data exists regarding responses of human atrial natriuretic factor (ANF) to exercise in the heat. The purpose of this study was to examine the responses of plasma ANF to high intensity submaximal (71% +/- 0.9 VO2max) exercise in the heat over an eight day acclimation period. Fourteen healthy males volunteered to participate in the study. Subjects performed intermittent exercises on a treadmill (0% grade) during 50 min of each 100 min trial in an environmental chamber maintained at 41.2 +/- 0.5 degrees C, 39.0 +/- 1.7% relative humidity. Blood was obtained from an antecubital vein after standing 20 min in the heat prior to exercise, and immediately after exercise. Measures were compared on days 1, 4 and 8. ANF did not change pre- to post-exercise nor did it change over the eight day heat acclimation period despite other heat acclimation adaptations. Conversely, plasma aldosterone (ALDO), renin activity (PRA) and cortisol (COR) all increased (p less than 0.05) pre- to post-exercise on each day but again no changes were observed over the eight day period. These data support that ANF may not increase when ALDO and PRA increases are observed. PMID- 2969333 TI - DNA-binding properties of an adenovirus 289R E1A protein. AB - An adenovirus 2 289 amino acid (289R) E1A protein purified from Escherichia coli has been shown to interact with DNA by two independent methods. UV-crosslinking of complexes containing unmodified, uniformly 32P-labelled DNA and purified E1A protein induced efficient labelling of the protein with covalently attached oligonucleotides, indicating that the E1A protein itself contacts DNA. Discrete nucleoprotein species were also observed when E1A protein--DNA complexes were analysed by gel electrophoresis. Although the 289R E1A protein exhibited no significant binding to single-stranded DNA or to RNA, no evidence for its sequence-specific binding to double-stranded DNA was obtained with either assay. Identification of the sites of covalent attachment of 32P-labelled oligonucleotides by partial proteolysis of the crosslinked E1A protein indicated that the interaction of this protein with DNA is mediated via domain(s) in the C terminal half of the protein. Such previously unrecognized DNA-binding activity is likely to contribute to the regulatory activities of this important adenoviral protein. PMID- 2969335 TI - Plasma levels of beta-endorphin, prolactin and gonadotropins in male athletes after an international nordic ski race. AB - Plasma beta-endorphin, prolactin (PRL), FSH and LH were measured in 17 volunteer male subjects at rest and under the stress caused by a long-distance nordic ski race. The race induced increased levels of beta-endorphin and PRL in all skiers. The changes in PRL with exercise were significantly related to the changes in beta-endorphin (r = 0.69, p less than 0.001). Furthermore, the highly trained skiers training over 150 km.week-1 of nordic ski showed consistently higher post exercise beta-endorphin and PRL levels than the moderately trained skiers who trained for 20 km.week-1. In addition the race induced slight falls in FSH and LH; however plasma gonadotropin levels did not show any correlation with plasma beta-endorphin concentrations and did not differ between the two groups of skiers. These results suggest that endogenous opioid peptides may modulate PRL secretion in heavy exercise, since they are of minor importance in the release of FSH and LH in such a situation. The observations also suggest that the degree of previous training and the exercise intensity do seem to be responsible for the hormonal changes. PMID- 2969336 TI - Partitional measurement of capillary and arteriovenous anastomotic blood flow in the human finger by laser-Doppler-flowmeter. AB - This study was made to see whether changes in blood flow through the capillaries and arteriovenous anastomoses (AVA's) of the human finger can be measured by noninvasive flowmetry. Total finger blood flow (FBF) was measured by venous occlusion plethysmography; blood flow was measured by a laser-Doppler flowmeter (ADVANCE, ALF-2100, Tokyo, Japan) using probes with optic fiber separations of 0.3 mm (LDF-0.3) and 0.7 mm (LDF-0.7). The maximum sensitivities for LDF-0.3 and LDF-0.7 were at depths of 0.8 and 1.2 mm from the tissue surface respectively. Two series of experiments were performed on separate days. In the first series the test hand was immersed in a water bath whose temperature (Tw) was 25 degrees C at an ambient temperature (Ta) of 25 degrees C. Tw was raised to 35 degrees C (local hand warming), which was then followed by an increase in Ta to 35 degrees C (whole body warming). FBF, LDF-0.3, and LDF-0.7 increased during these thermal stimulations. However, the relationship of FBF to LDF-0.3 showed two different regression lines. In contrast, the relationship of FBF to LDF-0.7 showed a single regression line. In the second series, with Ta at 35 degrees C, the test hand was immersed in a water bath at Tw 35 degrees C. Tw was then raised every 10 min by 2 degrees C steps from 35 to 41 degrees C. At Tw 39-41 degrees C, FBF and LDF-0.7 in the test hand were significantly decreased compared with those at Tw 35 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969337 TI - Induction of multiple germ tubes in Neurospora crassa by antitubulin agents. AB - The antitubulin fungicide benomyl suppressed the linear growth of Neurospora crassa wild type strain St. Lawrence 74 at micromolar concentrations. The rate of germination of macroconidia was not affected. Macroconidia exposed to 1.7 microM benomyl for 5 h formed multiple germ tubes. When germlings incubated for 4 h were exposed to 1.7 microM benomyl for 3 h, their germ tube stopped growing, swelled and emitted several branches. Normal linear growth was restored after removal of the fungicide. Linear growth of N. crassa was resistant up to 16 microM nocodazole. This drug induced multipolar germination at 8 microM, and griseofulvin only at 140 microM. The microtubule (MT) cytoskeleton of N. crassa could be revealed by indirect immunofluorescence with the monoclonal antibody YOL 1/34 directed against yeast alpha-tubulin. We detected no striking effects of the benomyl treatments on MT organization. The MT-stabilizing agents deuterium oxide (D2O) and cAMP have no antagonistic effects on the benomyl-induced multipolar germination. The positioning of nuclei and mitochondria was determined from the DAPI and Rhodamine 123 fluorescence patterns, respectively. Benomyl inhibited nuclear migration into multiple germ tubes. Quantitative scanning cytophotometry revealed a peak in the intensity of the mitochondria-associated Rhodamine 123 fluorescence near the apex of untreated germlings. This peak disappeared in multiple germ tubes. Benomyl-resistant mutant bml 511 (r), mutated in its beta tubulin gene, germinated normally in the presence of the fungicide. This strongly suggests that multiple germ tube formation was due to the effect of benomyl on beta-tubulin. Benomyl-resistant strain 74-3, constructed by reintroducing the cloned mutant N. crassa beta-tubulin gene into the cells by transformation, displayed a partial resistance to benomyl with respect to multipolar germination. Its rate of germination was slow (50% germination reached after 4 h at 37 degrees C as compared to 2.5 h for the wild type). In contrast to N. crassa, the other ascomycete Aspergillus nidulans is nocodazole-sensitive (linear growth suppressed at 1.6 microM). It did not respond to the MT inhibitors benomyl and nocodazole with respect to the pattern of germ tube emergence. Our results suggest that microtubule or membrane beta-tubulin is involved in the maintenance of developmental polarity during germ tube emergence and growth of N. crassa. PMID- 2969338 TI - Serotonin involvement in the inhibition of luteinizing hormone (LH) release induced by prostaglandin D2 (PGD2) in castrated male rats. AB - p-Chlorophenylalanine (PCPA, 320 mg/kg i.p.), an inhibitor of serotonin (5-HT) synthesis, and 5,6-dihydroxy-tryptamine (5,6-DHT, 50 micrograms i.c.v.), a drug toxic to the indoleaminergic system were used to test the involvement of 5-HT in the mediation of the inhibitory effect of prostaglandin D2 (PGD2) on luteinizing hormone (LH) release in castrated male rats. The i.c.v. administration of PGD2 suppressed the episodic LH release characteristic of castrated rats and decreased mean plasma LH levels and mean LH pulse amplitude significantly. Pretreatment with PCPA or 5,6-DHT apparently eliminated the inhibitory effect of PGD2 on LH secretion. These results suggest the possible involvement of a serotonergic mechanism in the mediation of the suppression of LH secretion induced by PGD2 in castrated male rats. PMID- 2969339 TI - Electrophysiological evidence that spiperone is an antagonist of 5-HT1A receptors in the dorsal raphe nucleus. AB - The neuroleptic spiperone, which binds to 5-HT1A, 5-HT2 and dopamine (DA) receptors, was studied for its effects on serotonin (5-HT) and DA neurons in dorsal raphe nucleus and substantia nigra pars compacta, respectively. We found that 1 mg/kg i.v. spiperone, but not LY53837 (a 5-HT2 antagonist), antagonized the inhibition induced by 5-HT1A agonists 8-hydroxy-dipropylaminotetralin (8-OH DPAT) and buspirone in the dorsal raphe nucleus. Lower spiperone doses blocked DA receptors in substantia nigra pars compacta, but did not affect 5-HT neurons. Doses of 8-OH-DPAT completely silencing dorsal raphe neurons were ineffective in substantia nigra pars compacta. However, buspirone antagonized DA receptors in substantia nigra pars compacta with doses similar to those depressing dorsal raphe neurons. It is concluded that spiperone is an antagonist of 5-HT1A receptors in the dorsal raphe nucleus. PMID- 2969340 TI - Vasomotion in normal and injured spinal cord. AB - Blood flow in the feline thoracic spinal cord was measured, using a laser-Doppler flowmetry. Rhythmic changes in the spinal cord blood flow were repeatedly observed with a frequency of about three to eight cycles per minute, unrelated to the respiratory or cardiac cycles. These changes were unaffected by hypertension induced by angiotensin II, but disappeared when systemic blood pressure was lowered by trimethaphan or when hypercapnia was induced. CO2 responsiveness of the blood flow and postocclusive reactive hyperemia were also examined, in both the normal and injured spinal cord. CO2 responsiveness was lost 1 h after a 500 gm/cm injury without rhythmic changes, but was partially regained 2 days later, at which time rhythmic changes were frequently observed. Postocclusive hyperemia apparently diminished 1 h after a 500 gm/cm injury, but reappeared 2 days later. Vasomotion in the normal and injured spinal cord is discussed. PMID- 2969341 TI - Androgen parameters in hirsute women: correlations with body mass index and age. AB - The recent work was designed for studying the effects of body mass index (BMI) and age on serum hormone values in hirsute women of reproductive years with nontumorous hyperandrogenism. Two hundred patients were studied in the follicular phase of the cycle. The serum levels of testosterone (T), androstenedione (A), or dehydroepiandrosterone sulfate (DHEAS) were not associated with BMI, whereas the T/sex hormone-binding globulin (T/SHBG) ratio was significantly correlated with BMI independently of the effect of age. All serum androgens were significantly negatively correlated with age, and the serum levels were significantly lower in women older than 41 years when compared with women aged 25 years or less. After correcting for the effect of BMI, there was also a negative correlation between the T/SHBG ratio and age. The correlations between serum androgen parameters were calculated. We conclude that already in reproductive years, the age of the patient has modulating effects on serum androgens, and it should be taken into account when studying the hormone profile of hirsute women. PMID- 2969342 TI - [ATPase activity in neurons and glial cells of the brain of the rat after REM sleep deprivation (cytophotometric study)]. AB - Modified Washstein and Meisel's techniques were used for quantitative estimation of ATPases activity in the rat brain. The specific activity of Na+,K+-ATPase and Mg2+-ATPase was higher in glial cells than in the neurons. The 24-hr REM-sleep deprivation resulted in an enhancement of the activity of ATPases in neurons of the hippocampus whereas the activity of the above enzymes lowered in the gliocytes. The findings can be due to the changes in excitability of the neurons and depolarization of glia during the sleep deprivation. PMID- 2969344 TI - [Modulation of the action of interferon-gamma by protein G]. AB - These studies were designed to investigate the characteristics of the intracellular second messengers induced by interferons (IFN-alpha/beta and IFN gamma) after receptor binding. Pretreatment of target cells with V. cholerae toxin, which is Known to activate the GTP-binding stimulatory protein (Gs), potentiated the action of IFN-gamma, but not of IFN-alpha/beta. By contrast, B. pertussis toxin, which is known to act on the GTP-binding inhibitory protein (Gi doesn't affect the action of both IFN) (Gi). Besides this forskolin and PGE1, known to increase intracellular cAMP levels, completely prevented antiviral state induction by IFN-gamma, but had no effects on IFN-alpha/beta. Altogether these results demonstrate that IFN-gamma transduction signal is mediated by a G protein with functional characteristics similar to those of the known Gs protein. PMID- 2969343 TI - Plasma membrane phospholipid content in non-insulin-dependent streptozotocin diabetic rats--effect of insulin. AB - The activity of (Ca2+ +Mg2+)-ATPase is impaired in kidney basolateral membranes from non-insulin-dependent streptozotocin-diabetic rats. To study the possible role of changes in membrane phospholipid content in the malfunction of this enzyme in kidney membranes of the diabetic animals, phospholipid (phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and sphingomyelin) content was measured in kidney and liver membranes obtained from non-insulin-dependent diabetic rats. Total phospholipid content was similar in liver and kidney membranes of diabetic and control rats (595 +/- 47 versus 624 +/- 29 in liver and 469 +/- 22 versus 458 +/- 17 nmol Pi/mg protein in kidney respectively). Phosphatidylethanolamine content in kidney and liver membranes of diabetic rats was lower than in control rats (87.7 +/- 1.8 versus 96.4 +/- 2.2 nmol Pi/mg protein, p less than 0.01 and 87.1 +/- 3.7 versus 101.8 +/- 3.5, p less than 0.02 respectively). Phosphatidylinositol content was higher in kidney (28.0 +/- 0.6 versus 23.9 +/- 2.1, p less than 0.02) but not liver membranes from diabetic rats. The in vitro direct effect of insulin on the phospholipid content in kidney membranes was also measured. Physiologic concentrations of insulin (718 pmol/l for 30 min) increased the phosphatidic acid content in membranes from control but not from diabetic rats by 34.2% (p less than 0.02). This rise was readily measurable after 3 min of exposure to insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969345 TI - [Teicoplanin in otorhinolaryngology]. AB - In this study the clinical microbiologic efficacy of a new antibiotic, Teicoplanin, has been reported. Teicoplanin was given to both male and female patients (200 or 400 mg/die doses) suffering from ORL infections. The microbial flora of the inflammatory focus was checked before and after antibiotic administration. Complete microbiologic eradication occurred in 13 cases of the 18 examined; in 3 cases was observed reinfection and in 2 cases the isolates were not sensitive to the antibiotic tested. PMID- 2969346 TI - Plasma atrial natriuretic factor in cirrhotic patients with ascites. Effect of peritoneovenous shunt implantation. AB - Human atrial natriuretic factor (ANF) levels were measured before and after peritoneovenous shunt implantation in 10 cirrhotic patients with ascites, in whom sodium retention is a major clinical problem. The mean preoperative plasma level of ANF was 82 ng/L (normal range, 5-80 ng/L). Peritoneovenous shunting resulted in a significant rise in plasma ANF to 308 ng/L (p less than 0.0025) immediately after operation. This was followed by a constant fall until the seventh postoperative day, when mean plasma ANF was still significantly elevated (149 ng/L) compared with the preoperative value (p less than 0.01). Three months after shunt implantation mean plasma ANF had returned to the preoperative level (75 ng/L). Mean sodium excretion increased from 2.6 mEq/h preoperatively to 10.2 mEq/h at the second postoperative day (p less than 0.025). No direct relationship was noted between changes in plasma ANF level and changes in urinary sodium excretion after shunt implantation. These data demonstrate an intact ANF release response to intravascular volume expansion in cirrhotic patients with ascites, but exclude ANF as the diminished natriuretic factor as proposed by the overflow theory of ascites formation. Sodium excretion and fluid retention seem to be the result of vascular underfill and fluid maldistribution, and hormonal changes are likely to be secondary to them. PMID- 2969347 TI - [Treatment of tubal pregnancy by local and systemic administration of prostaglandin. Initial experiences]. AB - The present study reports on the treatment of 6 tubal pregnancies established by laparoscopy intratubal PGF2 alpha injection accompanied PGE2 applied systemically. The method was successful in all cases avoiding the usual surgical procedure. Concerning the side effects one patient exhibited higher postoperative temperature, whilst another showed hypertony and tachycardia immediately after PGF2 alpha injection which lasted a few minutes. Further studies should clarify whether this technically simple procedure is superior to the existing methods. PMID- 2969348 TI - Ventricular myosin from young and adult animals with respect to the thyroid state. AB - Studies were conducted to analyze the effect of the thyroid hormone on ventricular myosin during ontogenesis of mice, rats and rabbits. Hypothyroidism was induced in mice and rats by administering propylthiouracyl in drinking water. Rabbits were made hyperthyroid by chronic administration of thyroxine. The change in the thyroid state of rats and rabbits influenced young and adult animals differently depending on whether V1 or V3 was the major ventricular isomyosin form present. Measurements of Ca2+-ATPase activity of myosins from young and old control animals and from animals with changed thyroid state showed that hypothyroidism in rats is associated with a greater decrease of myosin ATPase in young rats which contain V1 isomyosin only, when compared with old rats which contain a preponderance of V3 isomyosin and less of the V1 form. In rabbits, ATPase activity of ventricular myosin was more elevated after thyroxine administration in adult rabbits, which contain V3 isomyosin only, than in young rabbits in which myosin consists of V1 and V3 isomyosins. Ventricular myosins of young and adult mice did not differ in their ATPase activity and the treatment of mice with propylthiouracyl had only slight effect on myosin ATPase. It can be concluded based on these results that the hypothesis concerning hypothyroidism inducing transformation of V1 into V3 isomyosin does not hold generally. PMID- 2969349 TI - [Bayesian estimates of unknown parameters of mathematical models of the dynamics of the mutation process and changes in the ratio of cells having passed different numbers of divisions in a culture]. AB - With the help of Bayesian methods, the conditions of solving experimental data samples and their divisions were established, equivalence of estimations of unknown linear dynamic models parameters proved, the estimations having been worked out by both accounting calculation errors and using their compensations with additional noise in the original model's discrete analog. The results are used in mathematical modeling of changing intensity of the process of hereditary pathology frequencies, and the process of changing the ratio of cells having passed different numbers of divisions in the culture. PMID- 2969350 TI - Smart2, a cosmid vector with a phage lambda origin for both systematic chromosome walking and P-element-mediated gene transfer in Drosophila. AB - We describe a new phage-lambda-replicon-based cosmid vector suitable for both chromosome walking and P-element-mediated transformation in Drosophila. Its unique BamHI cloning site is flanked by the promoters for the SP6 and T7-encoded RNA polymerases, permitting the synthesis of probes complementary to the ends of the cloned inserts for library screening. The selectable marker is tet for bacterial cell transformation and neo for Drosophila transformation expressed under the control of the Drosophila hsp70 promoter. PMID- 2969351 TI - Structure of an unusual sea urchin U1 RNA gene cluster. AB - Genomic clones containing multiple copies of the Lytechinus variegatus U1 gene have been isolated from a gene library in the phage lambda EMBL3. These clones contain both types of U1 RNA gene repeats interspersed in the same 15-kb fragment. In addition, about 1/3 of the repeat units contain a 260-bp insert 460 bp prior to the first nucleotide of the U1 RNA sequence. The inserted sequence is abundant in the sea urchin genome as judged by Southern blots of genomic DNA. There are no repeated sequences flanking the insert. The insert occurs at the same position in the highly conserved 5'-flanking region at which a deletion has previously been reported. PMID- 2969352 TI - Molecular cloning and expression in Escherichia coli of the cDNA coding for rat lipocortin I (calpactin II). AB - Lipocortins (LC) are a family of proteins that were initially described to be induced by glucocorticosteroids and to inhibit phospholipase A2 (PLA2). Using oligodeoxynucleotide probes corresponding to partial amino acid (aa) sequences of rat lipocortin I (LCI), we have isolated a cDNA clone for rat LCI from a cDNA library prepared from poly(A)+RNA of peritoneal cells of dexamethasone-treated rat. The cDNA insert (1355 bp) had an open reading frame of 1038 bp that encoded a 346-aa polypeptide (Mr 38,784). The nucleotide sequence and the amino acid sequence deduced from it showed high homology with the reported sequences of human LCI. A plasmid containing the trc promoter and cDNA sequence for 346 aa residues of the rat LCI was constructed and expressed in Escherichia coli. Antibody to human LCI crossreacted with the recombinant rat LCI, and the recombinant protein had characteristics of natural rat LCI including PLA2 inhibitory activity in vitro. PMID- 2969353 TI - [Hygienic assessment of working conditions for workers in coastal gas-separation stations]. PMID- 2969354 TI - [The work experience of a health service in the development and realization of a comprehensive health program]. PMID- 2969355 TI - [The work experience of the Byelorussian Health and Hygiene Research Institute in rendering practical aid to public health]. PMID- 2969356 TI - [Experience with improving working conditions in the heating chamber of the electric power plant]. PMID- 2969358 TI - [Plasma and urinary beta-thromboglobulin as indicators of platelet activation in pre-eclampsia]. PMID- 2969357 TI - Alteration of H2 receptor sensitivity in duodenal ulcer patients after maintenance treatment with an H2 receptor antagonist. AB - The effects of a specific H2 receptor agonist impromidine, on gastric acid secretion were measured in six patients with duodenal ulcer in clinical remission before and after three months treatment with ranitidine 150 mg nocte. After treatment basal acid output increased from 1.2 to 2.8 mmol/h and after maximal impromidine stimulation from 36.9 (4.7) to 44.2 (6.2) mmol/h (p less than 0.02). Intravenous ranitidine 50 mg was given at the end of the impromidine infusion on each study day; the antisecretory effect of intravenous ranitidine was accentuated after the treatment with ranitidine from a trough acid output of 8.5 (1.2) mmol/h before, to 3.8 (1.5) mmol/h (p less than 0.05) after, treatment. The increased response to the H2 agonist impromidine and the H2 antagonist ranitidine after treatment with ranitidine suggests an enhanced sensitivity of the H2 receptor. This might be explained on the basis of an increase in the number of H2 receptors ('up-regulation'). PMID- 2969359 TI - [Soluble T4 molecules inhibit HIV infection in vitro]. PMID- 2969360 TI - [AIDS. Trimetrexate--alternative in Pneumocystis carinii pneumonia]. PMID- 2969361 TI - [Alternatives to balloon dilatation]. PMID- 2969363 TI - Effects of modulating neonatal progestins and androgens on the development of play and other social behavior in the rat. AB - Rough-and-tumble play is sexually dimorphic in many species of mammals. Effects of androgen exposure on this behavior have been well documented, although the possible role of other hormones is less well known. This paper reports experiments in a series designed to investigate effects of neonatal exposure to progestins on the development of juvenile play behavior in rats. Specifically, the effects of altering levels of progestins and androgens in combination on the subsequent development of play behavior was studied, in order to investigate further the mechanism of progestogenic effects. We have previously [L. I. A. Birke and D. Sadler (1984). Physiol. Behav. 33, 217-219] reported an apparently anomalous effect of lowering endogenous progesterone using the antiserum given postnatally; this was predicted to raise levels of juvenile play. The prediction was, however, supported only for males; treated females showed lower levels of play than controls. The present study was designed to investigate these effects further, and to test hypotheses about the mode of action of neonatal progestins. Using various combinations of neonatal progestins (progesterone or medroxy progesterone acetate, MPA), and testosterone or the antiserum to testosterone, we analyzed the amounts of play and other social behavior shown by juvenile rats. The data support a hypothesis that the previously reported effects of progestin exposure on play result from weakly androgenic actions of the progestins, which presumably compete with other androgens in males. The cellular mechanisms involved are, however, unknown. PMID- 2969362 TI - Age-related changes in plasma dehydroepiandrosterone sulphate, cortisol, testosterone and free testosterone circadian rhythms in adult men. AB - The circadian rhythms of serum luteinizing hormone, follicle-stimulating hormone, testosterone (T), free testosterone (fT), sex hormone-binding globulin (SHBG), oestradiol, cortisol and dehydroepiandrosterone sulphate (DHA-s) have been investigated in 5 normal male adults and 6 elderly men. Circadian rhythms were detected statistically significant (p less than 0.05) by population mean cosinor analysis, for T, fT, cortisol and DHA-s in the young group. In the elderly population, serum cortisol showed a clear circadian rhythm, although with some phase modification, whereas DHA-s secretion lost its circadian rhythmicity. This demonstrates that ageing differently affects the two major adrenal functions, glucocorticoid and androgenic; further, the data suggest that an independent adrenal androgen-regulating system could be selectively impaired in the older subjects. In the elderly group the loss of T circadian rhythm was confirmed, but a statistically significant circadian rhythm of fT was recorded. It was characterized by a marked phase advance and not related with the SHBG modifications found in elderly men. This finding leads us to reconsider the role of fT, which appears more sensitive than total T, in studying circadian rhythm of gonadal androgen secretion. PMID- 2969364 TI - Gc and PGM subtypes in northern Greece. AB - By isoelectric fucusing, Gc and PGM subtypes were examined in a sample of over 450 Greeks from Thessaloniki and surrounding areas. The gene frequencies are compared with those from other Greek and European samples. PMID- 2969365 TI - Immunocytological and immunochemical analysis on the common acute lymphoblastic leukemia antigen (CALLA): evidence that CALLA on ALL cells and granulocytes are structurally related. AB - The common acute lymphoblastic leukemia antigen(CALLA) on acute lymphoblastic leukemia(ALL) cells and granulocytes were compared by newly developed and other anti-CALLA monoclonal antibodies(anti-CALLA). New anti-CALLA(IF-3 through IF-7) were effectively selected by immunostaining on kidney sections. By competitive binding three antigenic determinants were separated on ALL cells by IF and other anti-CALLA. All three determinants existed on granulocytes although the reactivity of each anti-CALLA was variable. Such a variability was partly due to the heterogeneous terminal sialic acid compositions. Although CALLA from granulocytes and ALL cells differed in molecular weight they showed identical peptide mapping patterns. These results strongly suggest that CALLA on ALL cells and granulocytes are structurally related although they are different in posttransulational modification. PMID- 2969366 TI - A time-shared ultrasound Doppler measurement and 2-D imaging system. PMID- 2969368 TI - Comparison of zero-crossing counter to FFT spectrum of ultrasound Doppler. PMID- 2969367 TI - Multifrequency Doppler flowmeter: a way to rapid CW Doppler imaging. PMID- 2969369 TI - In vitro effects of ethanol and DDT on myofibrillar-ATPase in cardiac and certain skeletal muscles of pigeon (Columba livia) and rat (Rattus rattus). PMID- 2969370 TI - A mouse T lymphoma that lacks T-cell receptor surface expression exhibits allelic exclusion of its beta chain genes. PMID- 2969371 TI - T-cell receptor gene rearrangement in primary tumors: effect of genetic background and inducing agent. AB - The status of T-cell receptor beta and gamma genes has been assessed in a series of primary tumors induced by a chemical carcinogen or by gamma-irradiation using two inbred strains of mice. It appears that these well-characterized regimens of carcinogenesis yield T-cell tumors showing gene rearrangements consistent with a clonal origin of the tumors. Individual rearranged bands seem to represent orthodox, intralocus recombination events. A variety of rearrangement phenotypes are observed, most strikingly for the gamma genes, and differences in the degree of T-cell receptor gene rearrangements observed can be categorized according to the inducing agent and to the genetic background of the mice, with the implication that premalignant thymocytes have been captured in different stages of T-cell development. Additionally, primary tumors were shown to express significant levels of mature beta gene mRNA. PMID- 2969372 TI - Effect of vasopressors on atrial natriuretic factor and hemodynamic function in humans. AB - To assess the effects of vasopressors on plasma levels of immunoreactive atrial natriuretic factor (ANF), 13 normal men were studied on two occasions. On the experimental day, subjects received sequential 15-minute intravenous infusions of angiotensin II in doses of 4, 8, and 16 pmol/kg/min. Following a 30-minute recovery period, subjects received sequential 15-minute infusions of phenylephrine in doses of 0.4 and 0.8 micrograms/kg/min. Right atrial pressure, mean pulmonary capillary wedge pressure, pulmonary artery pressure, mean systemic arterial pressure, and plasma levels of renin activity, aldosterone, angiotensin II, and immunoreactive ANF were obtained sequentially throughout the protocol. During the control day, vehicle was infused and plasma samples were obtained for hormone measurements. Infusion of angiotensin II and phenylephrine increased mean systemic arterial pressure in a stepwise fashion. Both right atrial pressure and pulmonary capillary wedge pressure increased significantly during both doses of phenylephrine, but only the highest dose of angiotensin II significantly increased atrial pressures. Plasma levels of immunoreactive ANF increased parallel with the changes in right atrial pressure and pulmonary capillary wedge pressure, with significant increases occurring only at the highest dose of both pressors. Angiotensin II and aldosterone levels increased and renin activity decreased during infusion of angiotensin II. There were no significant changes in plasma levels of immunoreactive ANF during the control day. These studies demonstrate that infusion of vasopressors increases plasma levels of ANF, but only when the vasopressor effect is associated with significant increases in right atrial and pulmonary capillary wedge pressures. Atrial stretch is the most likely mediator of the increase in plasma levels of immunoreactive ANF during vasoconstriction. PMID- 2969373 TI - T-cell subsets in delayed-type hypersensitivity, protection, and granuloma formation in primary and secondary Listeria infection in mice: superior role of Lyt-2+ cells in acquired immunity. AB - Immunity to Listeria monocytogenes was studied in mice treated with rat monoclonal antibodies (MAbs) specific for the Thy-1.2, L3T4, and Lyt-2 T-cell markers. Three characteristic T-cell-mediated phenomena were investigated. Delayed-type hypersensitivity (DTH) to listerial antigen was totally abolished in mice treated with anti-Thy-1.2 or anti-L3T4 MAbs, whereas anti-Lyt-2 MAb treatment had no effect, regardless of whether the MAb was given during the induction or the expression of DTH. On the other hand, the elimination of bacteria from the spleens of infected animals was inhibited only by the application of either anti-Thy-1.2 MAb or anti-Lyt-2 MAb. This could be shown most impressively during the secondary infection of immune mice with a normally lethal dose of listeriae. In this situation, treatment with anti-Lyt-2 MAb sufficed to completely abolish immunologic memory, whereas anti-L3T4 MAb had only a marginal effect on antibacterial protection. However, the accelerated development of mononuclear cell foci in the livers of immune mice was inhibited by the application of both anti-L3T4 MAb and anti-Lyt-2 MAb. It is concluded that in murine listeriosis, DTH and acquired immunity to reinfection are dissociable phenomena. Although DTH is a function of L3T4+ T lymphocytes, Lyt-2+ T cells are necessary and sufficient for the expression of acquired resistance to L. monocytogenes. The roles of the different T-cell subsets in granuloma formation warrant further investigation. PMID- 2969374 TI - Candida albicans C3d receptor, isolated by using a monoclonal antibody. AB - Pseudohyphae of Candida albicans possess a receptor for C3d, a fragment of the complement component C3. This receptor was partially purified by using a monoclonal antibody (CA-A) that previously had been shown to inhibit the binding of C3d to C. albicans pseudohyphae. Purified immunoglobulin G from ascites fluid (CA-A) was coupled to a cyanogen bromide-activated Sepharose column, and an affinity-purified fraction (A2) from C. albicans pseudohyphae was obtained. This fraction inhibited rosetting of the EAC3d receptor by pseudohyphae and appeared to contain glycoprotein, since receptor activity could be removed when A2 was incubated with lectins specific for mannose and glucose. A2 was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and two polypeptides of approximately 60 and 70 kilodaltons (kDa) were consistently identified in reducing gels. The 60-kDa protein was identified as a glycoprotein by concanavalin A binding. A2 was further analyzed by high-pressure liquid chromatography (HPLC). Of three fractions obtained by HPLC, one containing the 60 kDa protein was found to have receptor activity. When analyzed by HPLC, this protein was found to contain mannose and glucose in approximately equal amounts. Both immunofluorescence and electron microscopy of pseudohyphae treated with CA-A identified A2 as a surface moiety. Thus, the C3d receptor of C. albicans, isolated with CA-A, is a glycoprotein of approximately 60 kDa. PMID- 2969375 TI - Isolation and characterization of the Streptococcus mutans gtfC gene, coding for synthesis of both soluble and insoluble glucans. AB - The intact gtfC gene from Streptococcus mutans GS-5 was isolated in Escherichia coli in plasmid vector pUC18. The glucosyltransferase activity expressed by the gene synthesized both low-molecular-weight water-soluble glucan and insoluble glucan in a primer-independent manner. Purification of the enzyme by procedures that minimize proteolytic digestion yielded a purified preparation with a molecular weight of 140,000. Insertional inactivation of the gtfC gene with a streptococcal erythromycin resistance gene fragment followed by transformation of strain GS-5 suggested that the gtfC gene product was required for sucrose dependent colonization in vitro. In addition, evidence for the presence of a third gtf gene coding for soluble glucan synthesis was obtained following the construction of mutants containing deletions of both the gtfB and gtfC genes. PMID- 2969377 TI - The electromyographic silent period is reduced in individuals at risk for Huntington's disease. PMID- 2969376 TI - Age-associated increase in the expression of T-cell antigen receptor gamma-chain gene in conventional and germfree mice. AB - To determine whether environmental antigens such as intestinal microflora contribute to expansion of the T-cell repertoire, age-related changes in the expression level of T-cell antigen receptor genes encoding gamma, beta, and alpha chains were compared in the lymphoid tissues of conventional versus germfree mice. Irrespective of the conditions of maintenance, an age-associated increase in the expression of the gamma-chain gene was evident in the thymus and spleen. Both conventional and germfree old mice (age, 40 weeks) had a relatively high proportion of Thy1+ L3T4- Lyt2- cells but a reduced level of Thy1+ L3T4- Lyt2+ cells in the thymus compared with their counterparts (age 8 weeks). The thymic dysfunction but not the stimulation by intestinal microflora may contribute to this age-related increase in gamma-gene transcripts in these tissues. On the other hand, an age-associated increase in the expression of gamma RNA was not evident in the mesenteric lymph nodes of germfree mice, although a remarkable increase in the gamma-chain gene messages was detected in the lymph nodes of the aged conventional mice. These results suggest that the expression of gamma RNA in cells of gut-associated lymphoid tissue is partly influenced by intestinal microflora. PMID- 2969378 TI - Surgical treatment of persistent intestinal fistula associated with abdominal evisceration. PMID- 2969379 TI - Alternate technique for cul-de-sac insufflation for laparoscopy. PMID- 2969380 TI - Losses to internal parasites in swine production. AB - Experimental infections of pigs with Strongyloides ransomi, Stephanurus dentatus, Ascaris suum, Oesophagostomum spp. or Trichuris suis at increasing levels generally decrease daily gain and increase feed to gain ratio linearly. At lower, subclinical levels of infection, the feed to gain ratio typically is increased 3% to 6% by an infection. Based on low-level experimental infections with A. suum, the economic loss to producers in the U.S. from increased feed to gain ratio is estimated at $155 million annually. PMID- 2969383 TI - The potential of islet transplants as a cure for human diabetes. PMID- 2969381 TI - Subendothelial extracellular-matrix heparan sulfate proteoglycan-degrading activity of human monocyte macrophages. AB - At the early stage of atherogenesis, circulating monocyte macrophages appear to adhere to the endothelial cell surface and migrate subendothelially to become foam cells. The mechanism of these macrophage-endothelial cell interactions was investigated. Adherent macrophages isolated from human blood were plated on [35S]O4-prelabeled extracellular matrix-coated dishes prepared from cultured porcine aortic endothelial cells. During incubation for 2-3 days at pH 7.4 either in the presence or absence of serum, macrophages solubilized the labeled extracellular matrix to a lower molecular weight component (Kav approximately equal to 0.5) than the materials (Kav = 0) released into the medium containing no cells. The degrading activity was not stored intracellularly but instead was found pericellularly, requiring continuous cell-matrix contact. Heparin (10 micrograms/ml) inhibited this degrading activity of macrophages. Degradation products were precipitated with cetylpyridinium chloride and were resistant to further digestion with alkali, pronase, or chondroitinase ABC, but were converted to further lower molecular weight fragments (Kav = 0.84) after nitrous acid digestion or heparitinase treatment. The intact glycosaminoglycan side chains determined by subjecting the extracellular matrix to cleavage with alkali or pronase were larger (Kav congruent to 0.20) than those of degradation products released by macrophages. These results suggest that the attachment and subsequent invasion of endothelial cells by monocyte macrophages may involve the production of extracellular-matrix heparan sulfate proteoglycan-degrading activity by these cells. PMID- 2969382 TI - Mammary gland Ca2+-binding (-dependent) proteins: identification as calelectrins and calpactin I/p36. AB - Calcium-binding (-dependent) proteins (CBPs) associated with the spreading of mammary epithelial cell cultures have been identified as various calelectrins and calpactins (p36). In immunoblot analysis, the CBPs of 30-36 kD and 68-70 kD variously react with different calelectrin and calpactin I monomer/p36 antisera. The same immunoreactive proteins were shown to be present in virgin mammary glands and collagen gel mouse mammary epithelial cell cultures. The mammary CBPs show extensive immunochemical relatedness; however, they fail to show cross reaction with antiserum to calpactin II (lipocortin) antiserum. These immunoreactive CBPs comigrate in electrophoresis with 35S-methionine-labeled CBPs isolated from mammary epithelial cell cultures. Unlike calmodulin, the mammary CBPs that correspond to calelectrins and calpactin I monomer/p36 are not stable to thermal denaturation. The mammary CBPs bind to epithelial cell membranes in a Ca2+-dependent manner and are differentially released from ruptured cells, compared with calmodulin, suggesting subcellular localization. Phenothiazine agarose and phenylagarose are equivalent in their ability to bind the mammary CBPs. Thus, mammary gland CBPs of 30-36 kD and 68-70 kD have been shown to be related or equivalent to the calelectrins and to calpactin I monomer/p36. Since these proteins are known to bind Ca2+, we conclude that the mammary gland CBPs are also Ca2+-binding proteins. The mammary gland CBPs are immunologically related and probably represent members of a larger family of related proteins. PMID- 2969384 TI - The role of vitrectomy in the diabetic patient. PMID- 2969385 TI - The effect of cyclosporine on native renal function in non-uremic diabetic recipients of pancreas transplants. AB - In order to assess the nephrotoxic effect of cyclosporine on native kidneys in pancreas transplant recipients, serum creatinine and creatinine clearance levels were determined before and serially after pancreas transplantation alone in 33 non-uremic Type I diabetic patients. The mean serum creatinine level before transplant was 1.0 +/- 0.3 mg/dl, and the values at 2 weeks, 6 months, 1 to 2 years, and greater than 2 years after transplant were, respectively, 1.5 +/- 0.6, 1.4 +/- 0.4, 1.4 +/- 0.5, and 1.5 +/- 0.2 mg/dl. Mean creatinine clearance level before transplant was 90 +/- 34 ml/min, and the values at the corresponding time points were 55 +/- 23, 62 +/- 22, 58 +/- 13, and 63 +/- 16. Thus, the mean changes in both parameters were approximately 50% in the immediate period after transplant, and there was no additional deterioration by these measures of renal function. The changes were somewhat greater than those reported in newly diagnosed diabetic patients receiving cyclosporine for immunotherapy, but all of the subjects had histologic evidence of diabetic nephropathy at the time of transplant. Although the stabilization of renal function is encouraging, damage to the kidneys may be silent and serial renal biopsies are needed to allow full assessment of the impact of cyclosporine on renal pathologic conditions. PMID- 2969386 TI - Prevalence of microalbuminuria in maturity onset primarily non-insulin-requiring diabetes mellitus: effect of disease duration, glycemic control, and mean systemic blood pressure. AB - Because of the frequency of late cardiovascular complications in maturity onset non-insulin-dependent (Type II) diabetes mellitus, there have been few studies regarding nephropathy in this patient population. The authors have analyzed the prevalence of microalbuminuria and the nature of clinical manifestations associated with elevated albumin excretion rate (AER) in a large population presenting with Type II diabetes. Among 318 patients studied during 1986, pathologically elevated 24 h AERs were found in 59%. The rate of microalbuminuria among 205 Type I diabetic patients screened during the same interval was 43%. AER was found to be positively correlated with duration of disease (p less than 0.0008) and metabolic control as determined by measurement of glycosylated hemoglobin (HbA1c) (p less than 0.002). There was only a modest agreement between AER and mean systemic blood pressure. The high prevalence of microalbuminuria in Type II diabetic patients and its known association with increased mortality emphasize the need for long-term follow up studies in order to clarify whether elevated AER in this patient population is predictive for overt diabetic nephropathy. PMID- 2969387 TI - A reappraisal of the influence of blood rheology on glomerular filtration and its role in the pathogenesis of diabetic nephropathy. AB - The basic assumptions concerning the mechanisms of normal glomerular filtration are discussed. Attention is drawn to blood rheologic changes that follow glomerular filtration and influence postglomerular blood flow adversely. It is proposed that the blood rheologic changes will increase the resistance to flow in the peritubular plexus commensurate with the dimensions of the capillaries and blood viscosity in accordance with the general principles of the Poiseuille formula, even though blood is a non-Newtonian fluid. For this reason, the conditions of flow in the plexus must be a determinant of intraglomerular capillary pressure. When blood rheology is abnormal, as in insulin-dependent diabetic patients, the abnormality will be amplified by glomerular filtration and it is suggested that the consequences will be manifest as problems of blood flow in the peritubular plexus. As the increase in postglomerular intravascular pressure needed to restore the rate of blood flow to normal necessitates dilation of the afferent arteriole and possibly more proximal vessels, such changes will result in an increase in intraglomerular pressure. The increase in pressure that increases filtration is therefore a direct consequence of abnormal blood rheology. This concept provides a basis for understanding the mechanism of diabetic proteinuria and for other proteinurias associated with abnormal blood rheology. A possible role for altered blood rheology in the pathogenesis of both focal and total glomerulosclerosis is discussed, and the potential benefits of agents that improve blood rheology are outlined. PMID- 2969388 TI - Cognitive distortion in rheumatoid arthritis: relation to depression and disability. PMID- 2969389 TI - Dental care of handicapped children by general dental practitioners. PMID- 2969390 TI - [Massive fibroma of the round ligament developing as an extraperitoneal abdominal mass]. AB - The authors report the exceptionally rare case of a huge myofibroma of the round ligament. The localization of the myoma was very unusual. It developed from the inguinal insertion of the round ligament; and the fibroma had grown in the abdominal wall between muscle and the peritoneum. Consequently the mass was both abdominal and extra-peritoneal. To our knowledge, such an association has never before been described. In the case described here, the mass was asymptomatic and the patient presented because of her increasing abdominal size. Examination revealed an abdominal mass that almost reached the umbilicus and resembled a four months pregnancy. Ultrasonography showed that the uterus was normal. The mass was a 15 cm by 15 cm tumour of heterogenous structure. As the ovaries were not visualized it was impossible to decide whether the tumour was ovarian, intestinal or peritoneal. Laparotomy was performed with a preliminary diagnosis of a solid ovarian tumour. It was then possible to localize the mass accurately and to define it. Histological examination showed that it was a leiomyoma. Tumours of the round ligament are very uncommon. Among these tumours leiomyomas are the most frequent. Endometriotic tumours come next. Various very rare tumours can also be found. Leiomyoma of the round ligament may arise from each portion of the ligament: abdominal, inguinal or Labium Majus. But both abdominal and extra peritoneal development from the inguinal insertion of the ligament is rare. They are usually single, and unilateral, and are found by chance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969391 TI - Staphylococcus aureus at a maternity ward. I. Colonization of mothers and neonates and survival of various S. aureus types in colonized individuals. AB - Colonization by Staphylococcus aureus, characterized by phage type or production of enterotoxins and toxic-shock-syndrome-toxin, was followed in 50 mothers and their babies. Types or groups of staphylococci predominating during the particular period at the maternity department were evaluated according to survival rates in the colonized subjects. S. aureus frequently colonized nipples at the start of lactation and was found regularly on umbilical stump and in eye and mouth corners of the babies. During the second stage of the study phage untypable staphylococci producing enterotoxin C (NT/C) strongly predominated. These microorganisms colonized in the greatest extent both mothers and neonates. The majority of NT/C complex originated probably from one or two clones characterized, among others, by high biological activity. PMID- 2969392 TI - Redistribution of subpopulations of peritoneal macrophages with Fc gamma receptors under the effect of an immunomodulator. AB - The heterogeneity of resident peritoneal macrophages and peritoneal macrophages was studied in different periods following oral administration of sodium nucleinate according to their ability to bind and phagocytize sheep erythrocytes opsonized by means of specific rabbit IgG. Using a mathematical method developed earlier, it has been possible to demonstrate that resident peritoneal macrophages can be divided into two subpopulations--actively and poorly binding macrophages but, after activation by sodium nucleinate--into three subpopulations. Fractionation of the macrophages according to their ability to adhesion within a temperature gradient has shown that the same peaks are traced in the fractions as in the overall pool of cells, but in different quantitative ratios. It has also been demonstrated that phagocytic activity is reduced in macrophages capable of adhesion to plastic at lower temperatures. PMID- 2969393 TI - Increased autoantibody production by NZB/NZW B cells in response to IL-5. AB - We previously demonstrated that B cells from NZB/NZW but not nonautoimmune mice secrete high levels of autoantibodies in response to factor(s) derived from type 2 Th cell (Th2) clones. Supernatants from type 1 Th cell clones, which contain a different set of lymphokines, were not stimulatory. In the present experiments, we attempted to define the active Th2 factor(s) and to better understand the cellular basis for the hyperresponsiveness. In response to optimal concentrations of supernatant (Th2-Sup), B cells from 3-mo-old NZB/NZW mice produced up to 40 fold greater amounts of IgM anti-DNA compared with unstimulated B cells, whereas BALB/c B cells produced levels only slightly above background. Although Th2-Sup contained large amounts of IL-4, comparable concentrations of rIL-4 alone did not stimulate NZB/NZW B cells. Furthermore, a blocking anti-IL-4 mAb did not prevent Th2-Sup-stimulated autoantibody production. Th2-Sup was fractionated by HPLC, and the stimulatory factor(s) was found in fractions known to contain IL-5 (also known as B cell growth factor II). Indeed, a highly purified preparation of IL-5 reproduced the effects of Th2-Sup by stimulating NZB/NZW B cells to produce high levels of IgM anti-DNA antibodies while enhancing production by nonautoimmune cells only slightly. In limiting dilution studies, NZB/NZW compared with BALB/c spleens contained a three- to four-fold greater frequency of DNA-specific B cells that were responsive to IL-5. Together, the results suggest a potential role for IL-5 in the pathogenesis of NZB/NZW autoimmune disease. PMID- 2969394 TI - Simultaneous production of IL-2, IL-4, and IFN-gamma by activated human CD4+ and CD8+ T cell clones. AB - In the present study, we have investigated the ability of human T cells to secrete IL-2, IL-4, and IFN-gamma. IL-4 and IFN-gamma were quantified with enzymatic immunoassays and IL-2 with a biologic assay by using the murine IL-2 dependent cell line CTLL-2. PBL, stimulated with Con A or with a combination of the phorbol ester 13-O-tetradecanoylphorbol-12-acetate and the Ca2+ ionophore A23187 secreted IL-2, IL-4, and IFN-gamma. The kinetics of the secretion of the three lymphokines was investigated with two CD4+ clones; one (GEO-2) that produced IL-2, IL-4, and IFN-gamma and another (HY640), that produced only IL-2 and IFN-gamma. Significant IL-2, IL-4, and IFN-gamma production was observed after only 8 h of activation. Maximal levels of IL-2 and IL-4 were found 20 h after the onset of the stimulation which subsequently decreased. In contrast, IFN gamma levels continued to increase in a period up to 40 h and then leveled off. In spite of these differences in secretion, the kinetics of accumulation of mRNA did not differ. The IL-2, IL-4, and IFN-gamma mRNA were detectable 2 h after stimulation and continued to accumulate for a period up to 20 h. In a series of 22 CD4+ clones, 21 were able to secrete all three lymphokines upon stimulation. Almost all CD8+ clones were able to produce IL-2 and IFN-gamma, but only six of the 23 CD8+ T cell clones secreted IL-4. In addition, five CD4+ (allo)antigen specific T cell clones were tested for IL-2, IL-4, and IFN-gamma secretion upon specific stimulation. Two alloantigen-specific and two tetanus toxoid-specific T cell clones secreted IL-2, IL-4, and IFN-gamma simultaneously, whereas one alloantigen-specific T cell clone secreted IL-2 and IFN-gamma, but not IL-4. A supernatant of the CD4+ T cell clone GEO-2, that contained high levels of IFN gamma and IL-4, was unable to induce the low affinity receptor for IgE, CD23, on a Burkitt lymphoma cell line. However, after separation of IL-4 from IFN-gamma by using HPLC, the IL-4-containing fraction-induced CD23, which could be blocked by the fraction that contained IFN-gamma and by a polyclonal rabbit anti-IL-4 antiserum. Finally, the partly purified IL-4, that was devoid of IL-2, promoted the growth of the clone GEO-2. PMID- 2969395 TI - Translocation of protein kinase C in rat basophilic leukemic cells induced by phorbol ester or by aggregation of IgE receptors. AB - Rat basophilic leukemic cells contain protein kinase C (PKC), 96 +/- 1% of which is located in the cytosol in the resting state. Phorbol ester (PMA), synergistically with calcium ionophore (A23187), caused 55% of the total PKC activity to associate rapidly with membranes where it remained for at least 20 min. When IgE-loaded cells were activated by Ag, maximally 30% of the cytosolic activity associated with membranes within 15 to 30 s, but most of this returned to the cytosol by 2 min. The small amount (3%) of PKC activity that remained associated with the membranes did so for at least 20 min but only if aggregation of the receptors was maintained. PKC translocation correlated with aggregation of receptors both at 30 s and at 10 min. However, only the translocation at 10 min and not that at 30 s correlated with receptor-induced exocytosis. In the absence of extracellular calcium (no exocytosis is observed), translocation at 30 s was diminished by 30% and at 10 min was completely absent. Cells depleted of PKC by 18-h treatment with PMA failed to degranulate in response to PMA and A23187 but responded partially (35%) when receptors were aggregated. We conclude that translocation of PKC is an early event that follows aggregation of IgE receptors but may not be essential for mediating the exocytotic mechanism induced by these receptors. PMID- 2969396 TI - Recombinant human IL-2 overcomes genetic nonresponsiveness to malaria sporozoite peptides. Correlation of effect with biologic activity of IL-2. AB - Current malaria vaccine strategies focus on subunit vaccines that contain one or a limited number of malaria Ag. However, there is widespread nonresponsiveness to many of these Ag probably resulting from Ir gene control. Using a congenic mouse model, we demonstrated that human rIL-2 (as an adjuvant) can overcome Ir gene controlled low immune responsiveness to peptide malaria Ag vaccine candidates [R32tet32, R32LR, and Th2R-NP (NANP)5NA] as determined by the antibody response, providing it is emulsified with the Ag during immunization. This effect is not caused by IL-2 merely acting as a foreign protein and stimulating noncognate help; it requires biologic activity of the IL-2, as determined by studying the effect of inactive rIL-2, which has minimal biologic activity but which has retained its antigenicity. IL-2 does not appear to be working by an effect on priming of specific Th, and IL-2 cannot overcome an Ir gene controlled low T cell proliferative response. IL-2 may have a role to play in human vaccine development where a high titer antibody response to a subunit vaccine is required. PMID- 2969397 TI - Superinduction of low affinity IgE receptors on murine B lymphocytes by lipopolysaccharide and IL-4. AB - Recent work in both the human and murine systems has demonstrated that IL-4 is capable of specifically inducing the synthesis of the low affinity receptor for IgE (Fc epsilon RII). In addition, in conjunction with LPS, IL-4 will induce IgG1 and IgE synthesis. To analyze the correlation between Fc epsilon RII induction and IgE secretion, Fc epsilon RII and IgE levels were measured by RIA on murine splenic B cells stimulated with LPS and IL-4 over 7 days of culture. Treatment with LPS and IL-4 gave a 20- to 50-fold (day 3) "superinduction" of Fc epsilon RII levels compared with a 3- to 5-fold induction with IL-4 alone; removal of IL 4 resulted in a rapid decline in Fc epsilon RII levels. The cells expressing high Fc epsilon RII levels were determined to be blasts. Superinduction of Fc epsilon RII occurs at 10 U/ml IL-4 and remains relatively constant in the range of 10 to 1000 U/ml. In contrast, with increasing IL-4, IgE levels increase, reaching microgram levels at day 7 with 300 U/ml IL-4. Triggering the cells with anti-Ig, as expected, gave no Ig secretion, and in addition, Fc epsilon RII superinduction by IL-4 and anti-Ig was not seen. PMA is known to block Ig secretion induced by LPS. Concentrations of PMA that totally abrogated IgE secretion had no effect on Fc epsilon RII superinduction, indicating that the latter phenomena can be separated from IL-4-induced Ig secretion. Superinduction also results in higher levels of Fc epsilon RII fragment release into the media. Thus, attempts were made to influence IgE secretion by adding additional purified Fc epsilon RII fragment to the culture. The purified fragment did not have a significant influence on IgE levels in this system. PMID- 2969398 TI - Differential effects of glucocorticoids on the proliferation of a murine helper and a cytolytic T cell clone in response to IL-2 and IL-4. AB - The proliferation of murine T cell clones can be supported by IL-2 or by IL-4. We present here evidence that glucocorticosteroids differentially affect these two pathways of proliferation. Dexamethasone (DEX) and other corticosteroids were observed to induce autocrine proliferation of the D10.G4.1 Th cell clone (D10) in the presence of the anti-clonotypic antibody 3D3. This effect was inhibited by the anti-murine IL-4 antibody 11B11, indicating that it is mediated by IL-4. Furthermore, on this cell line, representative of the Th2 group of helper cells, DEX had little effect on the proliferation induced by exogenous IL-4 but completely inhibited the growth-promoting effects of IL-2. In contrast, the effects of DEX on the proliferation of the cytotoxic IL-2-dependent CTLL-2 cell line are completely opposite. DEX blocked the IL-4-driven proliferation of CTLL-2 cells, while leaving unaffected their response to IL-2. It is also shown in this study that the effects of glucocorticoids in this system are totally antagonized by the high affinity anti-glucocorticosteroid RU 38486, indicating that they are mediated through the described intracellular glucocorticoid receptor. These data suggest that the growth effects of IL-2 and IL-4 may be mediated by distinct pathways that are strikingly different in their sensitivity to glucocorticoids. In addition, the regulation of lymphokine-dependent proliferation and the response to glucocorticoids appeared very different in helper and cytotoxic cells. PMID- 2969399 TI - Ligand-loaded but not free complement receptors for C3b/C4b and C3d co-cap with cross-linked B cell surface IgM and IgD. AB - We have performed experiments to investigate possible physical interactions between C receptors (CR) and surface Ig (sIg) on the B cell plasma membrane. These molecules were found to be independent, non-linked, B cell surface structures, because capping CR1, CR2, sIgM, or sIgD with a specific antibody did not affect the distribution of the remainder of these molecules. Both CR1 and CR2, if bound by antibodies that did not independently cap CR, however, became associated with cross-linked sIg because CR that have been bound by intact anti CR antibodies or their Fab fragments co-capped with sIgM or sIgD that had been bound by divalent anti-IgM or anti-IgD antibody. CR1 that had bound C3b similarly co-capped with sIg when sIg was cross-linked. Ligand-bound or even cross-linked CR did not associate with non-cross-linked sIg because sIgD, bound by a univalent Fab fragment of anti-IgD antibody, did not co-cap with CR that had been cross linked by a sandwich of mouse anti-CR antibody and goat anti-mouse Ig. Other surface molecules, such as B1 and HLA-DR Ag, when bound by specific antibodies, did not cap with cross-linked sIg, and sIgD, when bound by a univalent Fab fragment of anti-IgD antibody, did not co-cap with cross-linked sIgM. Interactions between CR and sIg were not mediated by an association with IgG FcR because co-capping of CR and sIg was observed when F(ab')2 fragments of both anti CR and anti-Ig antibodies were used. These results demonstrate that B cell surface CR can become associated with sIg, but only if sIg is cross-linked and CR is bound by anti-CR antibody or has bound its natural ligand. PMID- 2969401 TI - Histological and histochemical study of nasal polypi. PMID- 2969400 TI - Regulation of Fc epsilon R2/CD23 gene expression by cytokines and specific ligands (IgE and anti-Fc epsilon R2 monoclonal antibody). Variable regulation depending on the cell types. AB - The regulation of human low affinity FcR for IgE (Fc epsilon R2/CD23) and the soluble Fc epsilon R2 [IgE binding factor (BF)] of monocyte (U937), T (ED), and B (JIJOYE) cell lines was examined by anti-Fc epsilon R2 mAb (H107, Mab176) and the cDNA probe for Fc epsilon R2. The effect of IL-4 and IFN-gamma on Fc epsilon R2 regulation was variable among these three cell lines. IL-4 and IFN-gamma enhanced the Fc epsilon R2 gene expression and the production of Fc epsilon R2 and IgE-BF on U937, whereas IL-4 and IFN-gamma had no significant effect on the Fc epsilon R2 expression on ED. On JIJOYE, IL-4 enhanced the Fc epsilon R2 and IgE-BF production on both protein and mRNA levels. In U937 and JIJOYE cells, there was a marked increase of Fc epsilon R2 mRNA after combined stimulation with IFN-gamma and IL-4. However, in JIJOYE cells, there was a dissociation between the surface expression of Fc epsilon R2 and Fc epsilon R2 mRNA treated with IFN-gamma plus IL 4. In these cells. IFN-gamma even down-regulated the IL-4-induced expression of surface Fc epsilon R2. Stimulation of JIJOYE cells with both IFN-gamma and IL-4 resulted in the increase of the IgE-BF in the supernatant, suggesting that IFN gamma enhanced the release of IgE-BF from Fc epsilon R2. The results indicated that Fc epsilon R2 and IgE-BF expression is regulated by IFN-gamma at least on two different levels: on transcriptional levels and the levels of cleavage of the surface Fc epsilon R2 to release soluble Fc epsilon R2 (IgE-BF). Ligands binding to the Fc epsilon R2 such as IgE and anti-Fc epsilon R2 mAb enhanced the surface expression of Fc epsilon R2 on these Fc epsilon R2(+) cell lines. This was mainly due to the surface accumulation of the receptors on JIJOYE and U937. However, the stimulation of ED by H107 and anti-Fc epsilon R2 mAb significantly enhanced the mRNA expression, indicating that Fc epsilon R2 synthesis may also be up-regulated by the specific ligands in some cell types. PMID- 2969402 TI - Alterations in Langerhans cells during growth of transplantable murine tumors. AB - To investigate the response of Langerhans cells to tumor growth, we examined the appearance and number of ATPase+ and Ia+ dendritic cells in the epidermis covering subcutaneous tumors. Mice were injected with cells from syngeneic UVB- and PUVA-induced tumors and a melanoma, and the overlying skin was examined at various times during progressive tumor growth. An increase in the number of ATPase+ and Ia+ dendritic cells was observed in skin over all three tumor types. Morphologic alterations in the cells were also noted, including a decrease in dendricity. These changes were apparent only in skin directly over growing tumor masses; contralateral and perilesional skin was unaffected. Injection of nontumorigenic cells and implantation of silicon did not induce changes in Langerhans cells. Regression of highly antigenic tumor cells and tumor regression in immunized mice were not accompanied by detectable alterations in Langerhans cells, whereas changes in Langerhans cells were apparent during tumor growth in nude mice. These results demonstrate that changes in the number and morphology of Langerhans cells occur in response to tumor growth but that the changes are not dependent on immunologic or inflammatory responses. PMID- 2969404 TI - [The concept of critical arterial stenosis: value of a model of measures obtained by an angiographic method and the Doppler technic before and after transluminal angioplasty of the iliac arteries]. AB - INTRODUCTION: Numerous studies in the literature have proven the fact that a critical size appears during the stenosis evolution. The work presented is a non invasive approach of this value, with correlation to the invasive results. The study was realized on a homogeneous group of 23 arteriopathic patients, selected for iliac artery angioplasty procedure. MATERIAL AND METHODS: Patients selection: twenty-three patients including 24 angioplasty procedures have been selected on a 21 month time study. Invasive data: two measurements are realized during angioplasty: trans-stenotic pressure gradient before and after angioplasty. stenosis calibration: both diameter and section have been calculated on radiological films. Non invasive data: a standard Doppler test is performed on lower limbs. Two parameters are compared with the invasive data: the ascending time of velocity curve at the common femoral level. the systolic pressure index (peripheral pressure/radial pressure). RESULTS: 1) Critical stenosis--invasive techniques (fig. 1): trans-stenotic pressure gradient versus stenotic measurements shaw an hyperbolic correlation. The critical stenotic level is found to be 60% in diameter and 80% in section. 2) Critical stenosis--non invasive techniques: stenotic measurements compared to ascending time (fig. 2) as well as pressure index at rest (fig. 3) shaw the same type of curve. The critical stenotic level is found to be the same than with invasive techniques. DISCUSSION: Some discordant results open the discussion about angioplasty as a "gold standard", specially with a geometrical analysis. Bad precision in non regular stenosis. Hemodynamic consequences of turbulent stenosis are not evaluated. Clinical interest of critical stenotic notion is underlined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969403 TI - Complement activation in sickle cell disease: a liposome model. AB - Patients with sickle cell disease (SCD) have poorly defined abnormalities of their alternative complement pathway (ACP). We have previously shown chronic activation of the ACP in these patients. To determine the mechanism of this finding, we studied concentrations of the complement control proteins factors I and H in serum from patients with SCD and found no significant difference when they were compared with a control population. Because certain membrane surfaces promote ACP activation and changes occur in erythrocyte membrane phospholipid organization with sickling, we used a liposome model to determine whether ACP activation could be caused by abnormal phospholipid organization of sickle cells. Liposomes with the composition of the sickle cell outer leaflet, which is enriched in phosphatidylserine and phosphatidylethanolamine, activated the ACP significantly more than liposomes with normal outer leaflet phospholipid content. Similarly, liposomes with the composition of the erythrocyte inner membrane leaflet, containing large amounts of phosphatidylethanolamine and phosphatidylserine, activated the ACP more than liposomes with the phospholipid content of the outer leaflet. These findings suggest that phospholipid composition of membranes may play a role in their ability to promote ACP activation, and that changes in phospholipid organization in sickle cells may contribute to the chronic ACP activation observed in patients with SCD. PMID- 2969405 TI - Helping patients decide: from Hippocrates to videodiscs--an application for patients with low back pain. AB - This paper outlines a modular, microcomputer and videodisc expert system, for patients with mechanical low back pain. The system incorporate facts, rules, and methods to extract data, opinions, information, and user preferences. It supports individual patient needs with a broad knowledge base, and analytical modelling techniques that connect flexible inference structures and user choices. Videodisc based expert systems help break the bottleneck in relevant medical knowledge representation. The expert system program serves the user in the capacity of teacher, confidant, interpreter, diagnostician, and adviser. PMID- 2969406 TI - Induction of interleukin 1 alpha mRNA during the antigen-dependent interaction of sensitized T lymphoblasts with macrophages. AB - DNA-RNA hybridization with an IL-1 alpha cDNA probe was used to monitor the induction of IL-1 in macrophages that were acting as accessory cells for the proliferation of T lymphocytes. Mouse peritoneal macrophages bound and stimulated T lymphocytes in the presence of the mitogens, Con A, or anti-CD3 mAb, but little or no IL-1 mRNA was detectable. In contrast, if the T cells were first sensitized in a mixed leukocyte reaction with dendritic cells and then added to macrophages, IL-1 mRNA was clearly induced. Induction of the IL-1 alpha gene seemed to require the recognition of class II MHC products on the macrophage because of the following observations: specific rather than third-party macrophages were responsive to the T blast but not to T cell-conditioned media; induction was blocked by an anti-Ia mAb; CD4+ rather than CD8+ blasts were active; and polyclonal Con A blasts were much less efficient than antigen-specific T cells. Our data indicate that the strongest signal for IL-1 production during the macrophage-T cell interaction occurs in the efferent limb of the response, after rather than before the formation of class II MHC-restricted T lymphoblasts. PMID- 2969407 TI - Medical disability assessment of the back pain patient for the Social Security Administration: the weighting of presenting clinical features. AB - We investigated how physicians use history and physical findings when assessing disability for low back pain. Thirty-six North Carolina physicians, either practitioners experienced in disability determinations (26) or employees of the Social Security disability agency (10), responded to 48 clinical vignettes. They rated each case on a scale of 0 to 1.0, according to their degree of certainty that the patient was disabled. All combinations of five patient variables were presented in the vignettes: pain (mild or severe), physical examination (normal, reflex loss or muscular weakness), mobility (normal or restricted), X-rays (normal or osteoarthritis), and occupational history (normal or light work). The mean certainties for the individual vignettes ranged from 0.08 to 0.43. Mean certainty estimates across physicians ranged from 0 to 0.61, indicating substantial variability in how physicians assess disability. Practicing physicians had higher certainty of patients' disability than did physicians employed by Social Security, 0.37 vs 0.07 (p less than 0.01). Degree of pain was not associated with certainty of disability. All other clinical factors were highly significant predictors of physician assessment of certainty of disability. The emphasis on physical and radiographic findings over history places disability evaluation distinctly apart from other medical assessments. PMID- 2969408 TI - Evaluation of the periodic examination in the South African mining industry. AB - The periodic examination in the South African mining industry was evaluated by determining the number and nature of abnormalities found in 7,758 consecutive examinations and the outcomes of these findings up to 3 years later. Separate analyses evaluated the examination in relation to its role as (1) a screening examination; (2) a means of assessing fitness for work, and (3) a means of identifying workers with compensable diseases. As a screening examination, very little benefit was realized, as only 132 examinations (1.7%) resulted in diagnoses considered significant according to specified criteria. As a means of assessing fitness for work, the examination appeared to be more useful. In 640 of the examinations (8.2%), conditions were discovered which were thought to have at least raised doubt about whether the certificate of fitness should be renewed unchanged. As a means of identifying workers with compensable diseases, the periodic examination was unproductive, with only 20 workers (0.26%) receiving compensation for conditions discovered during the periodic examination. It is suggested that the time between examinations can be extended, especially for the younger workers, and that follow-up of significant conditions can be undertaken more effectively if the examinations are conducted at the workplace. PMID- 2969409 TI - Schistosoma mansoni and S. japonicum worm numbers in 129/J mice of two types and dominance of susceptibility in F1 hybrids. AB - In a study on the genetics of resistance to schistosomiasis in WEHI 129/J mice, susceptibility to either Schistosoma mansoni or Schistosoma japonicum was shown to be unequivocally dominant in F1 hybrid crosses between genetically resistant WEHI 129/J and susceptible BALB/c mice. The operation of only 1 or 2 genes in the expression of resistance to S. mansoni was suggested by backcross analysis. Thus, approximately 25% of (BALB/c x WEHI 129/J) F1 x WEHI 129/J mice were resistant to S. mansoni infection, whereas resistance was manifest in approximately 50% of WEHI 129/J mice. The data are consistent with resistance being controlled by 1 recessive gene having 50% penetrance. We also report that 129/J mice obtained directly from the Jackson Laboratories (Bar Harbor, Maine) (designated JAX 129/J), differ from locally bred WEHI 129/J in being entirely susceptible to S. mansoni infection. However, both WEHI 129/J and JAX 129/J are relatively resistant to S. japonicum infection. PMID- 2969410 TI - Nutritional copper deficiency in severely handicapped patients on a low copper enteral diet for a prolonged period: estimation of the required dose of dietary copper. AB - Six cases of nutritional copper deficiency were identified in a nursery institute for severely handicapped patients. All had been on prolonged enteral feeding of a copper-deficient diet and all had abnormalities related to the copper deficiency. Two of them had leukopenia, with or without macrocytic and normochromic anemia. After the oral administration of copper, the clinical and laboratory observations including neutrophil counts and serum copper and ceruloplasmin levels showed a complete recovery. The dietary copper requirement was estimated to be approximately 20 micrograms/kg/day for these patients, based on the correlation between the dietary copper intake and the levels of copper in the serum. PMID- 2969411 TI - [Precordial ST-segment elevation caused by right coronary artery occlusion]. AB - Among 57 consecutive patients undergoing percutaneous transluminal coronary angioplasty (PTCA) of the right coronary artery, eight patients showed precordial ST-segment elevation in leads V1-3 during the procedure. The mechanism of this ST elevation was investigated reviewing the coronary angiographic findings. All patients had angina pectoris, but none had evidence of myocardial infarction. The balloon inflation time was limited to 60 sec, and 12 lead electrocardiograms were recorded every 15 sec. In the eight patients who had precordial ST-segment elevation, six had the anatomically dominant right coronary artery, and two had proportioned (balanced) left and right coronary arteries. Six patients, however, had functionally dominant left coronary arteries because of good collaterals supplying the right coronary artery from the left coronary artery. Thus, functionally, six had the dominant left coronary artery, one had proportioned coronary supply, and only one had the dominant right coronary artery. In all eight patients, the most proximal portion of the right coronary artery was occluded during PTCA, obstructing both the conus branches and the right ventricular branches. This often induced precordial ST-segment elevation in cases with the functionally dominant left or proportioned coronary artery. This ST segment elevation seemed to represent right ventricular ischemia, as the inferior wall was protected from ischemia by good collaterals. However, precordial ST segment elevation was rare in the functionally dominant right coronary artery even when the most proximal portion of the right coronary artery was occluded. This fact seemed due to masking of electrocardiographic manifestations of right ventricular ischemia by the dominant electrical forces of inferior wall ischemia. PMID- 2969412 TI - [Relationship between left ventricular diastolic behavior and the A wave ratio by the apexcardiogram: a study with echocardiography and pulsed Doppler echocardiography]. AB - We investigated the relationship between parameters of left ventricular diastolic filling using pulsed Doppler echocardiography and the A wave ratio of apexcardiography (ACG), and then evaluate the characteristic features of diastolic behavior in hypertrophic hearts and in various cardiac diseases. The study population consisted of 68 patients and 25 normal subjects, and included 19 cases of chronic renal failure (CRF), 17 cases of ischemic heart disease (IHD), 16 cases of hypertension (HT), six cases of hypertrophic cardiomyopathy, two cases of aortic stenosis, two cases of arrhythmias, and six of other cardiac diseases. The A wave ratio of ACG was calculated as the ratio of A wave amplitude and total excursion [(A/E-O) x 100]. At the same time, the peak early filling velocity (R), the peak late filling velocity (A), the ratio of R to A (A/R), acceleration time (AT), and deceleration time (DT) were measured from the left ventricular inflow velocity pattern using pulsed Doppler echocardiography. The results were as follows: 1. There was a close positive correlation between the A wave ratio of ACG and the A/R of pulsed Doppler echocardiography. 2. In patients with left ventricular hypertrophy (LVH), both the A wave ratio and the A/R were significantly higher than those in normal subjects. And in LVH with asynergy, both the A wave ratio and the A/R were significantly higher than those in LVH without asynergy. 3. In CRF, IHD, and HT, both the A wave ration and the A/R were significantly higher than those in normal subjects, but there were no significant differences among these three disease entities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969413 TI - [A report of the survey on the present application of percutaneous transluminal coronary angioplasty in Japan]. AB - To elucidate the current use of percutaneous transluminal coronary angioplasty (PTCA) in Japan, questionnaires concerning PTCA experience in 1986 and indications for using PTCA were sent to 53 relatively experienced centers. The results were collected from 46 centers (87%), and the data were analyzed. A total of 5,255 PTCA were performed in 1986, which was 35% greater than that in 1985. Of these, 4,440 PTCA procedures were performed in patients with angina. Primary success was achieved in 3,790 patients (85%), and complications included 21 deaths (0.5%), 41 emergency bypass surgery (CABG) (0.9%), and 118 myocardial infarction (MI) (2.7%). Another 815 PTCA procedures were performed in patients with acute myocardial infarction, of which 58% were direct PTCA and 42% were PTCA following thrombolysis. Primary success was achieved in 674 patients (83%) with complications of deaths in 19 (2.3%) and emergency CABG in 17 (2.1%). The initial treatments in patients with significant coronary stenosis diagnosed by initial coronary arteriography in 1986 were analyzed. In 6,845 patients with angina or old myocardial infarction, medical treatment was selected in 46%, PTCA in 40% and CABG in 14%. In 1,506 patients with acute infarction, medical treatment was performed in 21%, PTCA including PTCA following thrombolysis in 44%, CABG in 3% and only coronary thrombolysis in 32%. Questions on criteria required to select patients for PTCA and clinical or angiographic indications for PTCA were also asked. The following conditions were not necessarily required in more than half of the centers: symptoms, refractoriness to medical treatment and surgical candidates. However, evidence of myocardial ischemia and viability of the lesion related myocardium were required as criteria in the majority of centers. The following clinical or angiographic features were not always indicated in more than two-third of centers: i.e. cardiogenic shock, and 90% or less stenosis in acute infarction, 75% stenosis in variant angina, patients over 80 years old, poor cardiac function with 29% or less ejection fraction, severely calcified coronary lesions, long segment lesions (greater than or equal to 2 cm), distal lesions, unprotected left main trunk lesions, total obstructions (of more than three months duration), and three vessel disease with two total occlusions and one discrete lesion. PMID- 2969414 TI - Words as barriers. PMID- 2969415 TI - Complement C3d,g/Epstein-Barr virus receptor density on human B-lymphocytes estimated by immunoenzymatic assay and immunocytochemistry. AB - The density of the C3d,g/Epstein-Barr virus receptor (CR2) on human mononuclear cells (MNC) isolated from CPD stabilized blood, was determined by comparing the concentration of CR2 antigen in extracts from Raji cells, with a known receptor density, with extracts of MNC. The concentration of CR2 was determined by a sandwich ELISA using two mutually non-inhibiting monoclonal anti CR2 antibodies (HB5 and OKB7). The percentage of CR2+ MNC was determined by an immunocytochemical staining method. Analysis of MNC from 20 healthy donors, revealed that the density of CR2 molecules varied from 3,600-7,700 on CR2+ cells, and that the percentage of CR2+ MNC had a mean of 13%. PMID- 2969417 TI - A computer-aided programme for promoting unsupervized activities for multihandicapped adolescents. AB - The present study was aimed at developing a computer-aided programme for promoting performance of unsupervized activities with two multihandicapped adolescents. Simple household and occupational tasks already familiar to the subjects were selected as activities. Both subjects discriminated the pictorial representations of those activities. In order to control for the effects of the computer-aided programme, the subjects were also exposed to a card programme. Data indicate that the computer-aided programme was successful with both subjects. Advantages and limitations of this programme are discussed. PMID- 2969416 TI - ATP-dependent calcium transport in rat parotid basolateral membrane vesicles is modulated by membrane potential. AB - The ATP-dependent Ca2+ transport activity (T. Takuma, B.L. Kuyatt and B.J. Baum, Biochem. J. 227:239-245, 1985) exhibited by inverted basolateral membrane vesicles isolated from rat parotid gland was further characterized. The activity was dependent on Mg2+. Phosphate (5 mM), but not oxalate (5 mM), increased maximum Ca2+ accumulation by 50%. Half-maximal Ca2+ transport was achieved at approximately 70 nM Ca2+ in EGTA-buffered medium while maximal activity required greater than 1 microM Ca2+ (Vmax = 54 nmol/mg protein/min). Optimal rates of Ca2+ transport were obtained in the presence of KCl, while in a KCl-free medium (mannitol or sucrose) approximately 40% of the total activity was achieved, which could not be stimulated by FCCP. The initial rate of Ca2+ transport could be significantly altered by preimposed membrane potentials generated by K+ gradients in the presence of valinomycin. Compared to the transport rate in the absence of membrane potential, a negative (interior) potential stimulated uptake by approximately 30%, while a positive (interior) potential inhibited uptake. Initial rates of Ca2+ uptake could also be altered by imposing pH gradients, in the absence of KCl. When compared to the initial rate of Ca2+ transport in the absence of a pH gradient, pHi = 7.5/pHo = 7.5; the activity was approximately 60% higher in the presence of an outwardly directed pH gradient, pHi = 7.5/pHo = 8.5; while it was approximately 80% lower when an inwardly directed pH gradient was imposed, pHi = 7.5/pHo = 6.2. The data show that the ATP-dependent Ca2+ transport in BLMV can be modulated by the membrane potential, suggesting therefore that there is a transfer of charge into the vesicle during Ca2+ uptake, which could be compensated by other ion movements. PMID- 2969418 TI - Short-term memory difficulties and Down's syndrome. AB - Nonretarded (NR) individuals typically show better short-term memory for brief sequences of auditory than visual information (the modality effect). The present study attempted to determine whether the failure of Down's syndrome (DS) individuals to show the modality effect is due to the verbal-expressive demands of oral responding in memory tasks. DS, NR and MR (non-DS mentally retarded) subjects listened to or looked at increasingly long sequences of digits and attempted to recall them either orally or manually (through placement of items). Analyses suggested the following: (1) manual responding failed to enhance auditory recall in either DS or any other subjects; and (2) difficulty in recalling auditory stimuli was greater for DS mentally retarded subjects. An additional assessment of DS, MR and NR subjects on a standardized auditory short term memory test requiring a nonverbal pointing response replicated the above findings. PMID- 2969419 TI - Tourettism in a patient with Down's syndrome. PMID- 2969420 TI - The effect of linoleic and arachidonic acid derivatives on calcium transport in vesicles from cardiac sarcoplasmic reticulum. AB - The effect of linoleic and arachidonic acid derivatives on ATP-dependent calcium transport was studied in the isolated vesicles from cardiac sarcoplasmic reticulum of guinea-pigs. Oxidation products of linoleic and arachidonic acids, obtained either by autoxidation or incubation with soybean lipoxygenase, effectively blocked in a dose-dependent manner, the net influx of calcium in the absence or presence of 5 mM of oxalate. Unoxidized fatty acids were much weaker at lower concentrations as compared to their oxidized counterparts, except the lipoxygenase-generated product of arachidonic acid which had only a marginal effect even at high concentrations. Autoxidation products of arachidonic acid were the most potent inhibitors of calcium transport. Likewise, autoxidation products of linoleic and arachidonic acids and lipoxygenase-generated products of linoleic acid induced a dose-dependent release of calcium from vesicles previously loaded with 45Ca, and release was further enhanced in the presence of 0.5 mM of EGTA. In contrast, lipoxygenase metabolites of arachidonic acid caused a transient increase in net calcium content. The effect of the fatty acid derivatives on calcium transport did not appear to be due either to the inhibition of Ca2+-ATPase activity or to a non-specific detergent-like action. The effects of oxidized fatty acids, on ATP-dependent calcium accumulation into and release from cardiac microsomal fraction were similar but less potent than those of classical calcium ionophores, X537A or A23187. PMID- 2969421 TI - Susceptibility of hypertrophied rat hearts to ventricular fibrillation during acute ischemia. AB - This study compared arrhythmias induced by acute ischemia in Langendorff preparations of normal and hypertrophied rat hearts. Left ventricular pressure overload was induced by partial ligation of the abdominal aorta 6 to 8 weeks prior to study. The ratio of left ventricular weight to body weight (LVW/BW) and systolic blood pressure (SBP) were significantly higher in two groups of rats with hypertrophied hearts (moderate hypertrophy: 2.68 +/- 0.06 mg/g, 182 +/- 3 mmHg; severe hypertrophy: 3.31 +/- 0.03 mg/g, 238 +/- 5 mmHg) than in normal hearts (2.18 +/- 0.03 mg/g, 125 +/- 3 mmHg), while there were no differences in body weights. During 30 min of ischemia produced by left coronary artery occlusion in the Langendorff preparations, ventricular fibrillation occurred in six of 20 (30%) normal, six of nine (67%) moderately hypertrophied, and 14 of 14 (100%) severely hypertrophied preparations (P less than 0.001 normal vs severely hypertrophied). Tachyarrhythmias occurred in 15 of 20 (75%) normal, eight of nine (89%) moderately hypertrophied, and 14 of 14 (100%) severely hypertrophied hearts. Heart rate and coronary efflux before and during the ischemic period did not differ between normal and hypertrophied hearts. The ratio of non-perfused to perfused areas of the left ventricle, measured by Evans blue dye staining and with computerized planimetry, also was not different for normal (57.6 +/- 2.3%) and hypertrophied hearts (moderate hypertrophy 62.5 +/- 3.3%, and severe hypertrophy 59.1 +/- 2.0%). Additional control studies using larger hearts from older rats also indicate that myocardial mass was not an important determinant of ischemic arrhythmias in hypertrophy. Prolongation of endocardial and epicardial conduction time during 30 min of ischemia was not different between normal and hypertrophied hearts. Action potential duration and refractory periods were significantly longer in ventricular cells of hypertrophied hearts than in normals, and superfusion with hypoxia/zero glucose solution shortened these parameters to a greater extent in hypertrophied cells. These results lead us to conclude that hypertrophied hearts have a greater susceptibility to ventricular fibrillation during acute ischemia, and that dispersion of refractoriness may play a role in this phenomenon. PMID- 2969422 TI - Hypertrophic and functional response to experimental chronic aortic regurgitation. AB - Aortic regurgitation was induced by retrograde perforation of an aortic valve cusp under hemodynamic guidance in 12 New Zealand White rabbits. Regurgitant fraction was documented by electromagnetic flow probe and six sham-operated animals served as controls. Two-dimensional, M-mode and Doppler echocardiography was performed pre-operatively and serially post-operatively for 3 to 6 months. Animals with aortic regurgitation developed progressive left ventricular dilatation and eccentric hypertrophy. Left ventricular internal dimension at end diastole and left ventricular mass were increased from baseline values by 41 and 94% (P less than 0.001), respectively; fractional shortening was stable while end systolic stress increased 50% (P less than 0.01. Thus, acutely induced aortic regurgitation in rabbits results in a chronic model which may be appropriate for stimulation of the hypertrophic response to aortic regurgitation in humans. PMID- 2969423 TI - Cardiac output determinations in the newborn. Reproducibility of the pulsed Doppler velocity measurement. AB - Cardiac output (QAo) can be estimated noninvasively by pulsed Doppler (PD) ultrasonographic determination of mean ascending aortic blood flow velocity (VAo) combined with M-mode echocardiographic determination of ascending aortic cross sectional area (AAo). Cardiac output is calculated from the volumetric flow equation (QAo) = (VAo) X (AAo). Pulsed Doppler measurements are known to correlate well with Fick and thermodilution methods; however, inter- and intraoperator variability of the velocity component of the PD method has not been determined in newborns. We did three repeated PD measures of mean aortic flow velocity in ten term infants (using four trained operators) to determine inter- and intraoperator reproducibility. The coefficient of variation for intraoperator variability (random error) for a single measurement of VAo was 11.7%. If three repeated measures by a single operator were averaged, the random error was 7.0%. There was little interoperator variability found. PMID- 2969424 TI - Transvaginal oocyte retrieval controlled by vaginal probe for in vitro fertilization: a comparative study. AB - Of 142 patients who initiated in vitro fertilization cycles, 52 underwent oocyte collection by laparoscopy, 58 by the ultrasonically guided transvesical route, both under general or local anesthesia and 36 by the ultrasonically transvaginal route under local anesthesia. When the three methods were compared the results did not differ significantly. An average of 6.4, 6.2, and 5.7 oocytes, respectively, were retrieved. Cleavage rates were 82.6, 79.4, and 81.6%, respectively, while 20.2, 22.6, and 21.1% clinical pregnancies per cycle were obtained. Complications with this procedure such as exacerbation of previous pelvic inflammatory disease, urinary tract infections, transient hematuria, and mild hemoperitoneum were rare. In general, patients greatly preferred the transvaginal method. In addition to providing greater comfort for the patient, this method requires no general anesthesia and is less expensive than other methods. Our initial experience demonstrates that in vitro fertilization can be performed successfully through this simplified alternative to oocyte retrieval. PMID- 2969425 TI - The fate of the donor artery in extraanatomic revascularization. AB - Favorable long-term results have been reported after femorofemoral bypass in the treatment of iliofemoral occlusive disease. Functional alterations in the donor artery, occurring after the implantation of the graft, have been considered as having an arresting effect on the progress of the occlusive disease, which explains the favorable long-term results. However, until now, the intrinsic mechanism of this phenomenon has remained unexplained. Little attention has been devoted to the evaluation of the natural history of the donor artery. To achieve this purpose, 18 patients previously operated on for femorofemoral bypass, with a normal patent graft, were submitted to angiographic evaluation, from 8 to 57 months after the operation (mean 22 months). The angiograms were compared with the preoperative films to assess the progress of the occlusive disease in the donor artery. A slight but significant dilatation of the proximal donor arterial segment was documented in every case, affecting the external iliac and common femoral arteries. The pathogenesis of this phenomenon is discussed. It may be related to decreased peripheral resistance and increased blood flow occurring in the donor arterial segment, reported after the implantation of the shunt. This dilatation of the donor artery may have a compensatory effect for local atherosclerotic stenosis, thereby explaining the favorably long duration of the femorofemoral bypasses, which has been emphasized since the early days of extraanatomic revascularization. A similar phenomenon may occur in the vessels of the upper extremity, after axillofemoral revascularization. PMID- 2969426 TI - Mechanisms of healing in synthetic grafts. PMID- 2969427 TI - Pathologic study of myocardial infarction of the right ventricle. AB - To evaluate pathologic features of myocardial infarction of the right ventricle (MI-RV), we analyzed 106 autopsy cases with transmural myocardial infarction (MI) (fresh in 46 cases and healed in 60). Anterior MI was observed in 47, posterior MI in 54 and lateral in 5. There were 13 cases (12%) with MI-RV (anterior in 1 case and posterior in 12), which included 10 cases with fresh MI and 3 with healed MI. All cases with MI-RV had associated transmural interventricular septal infarction. Of the 13 cases with MI-RV, 9 (69%) had right ventricular dilatation (RVD) and 2 had right ventricular hypertrophy. Extensive MI-RV (more than 1/3 of the right ventricle) was observed in 8 (89%) of those with RVD. Of 93 cases of MI without MI-RV, 14 (16%) had RVD. The incidence of RVD was greater in cases with MI-RV than in those without (p less than 0.005). All 12 cases with posterior MI RV had significant (greater than or equal to 75%) narrowing of the right coronary artery (RCA), and 19 cases (87%) of those with posterior MI without MI-RV, had similar lesions. In conclusion, the incidence of RVD and significant narrowing of RCA was greater in cases with posterior MI-RV than in those with posterior MI. PMID- 2969428 TI - Acute hemodynamic effects of alpha human atrial natriuretic polypeptide in patients with congestive heart failure. AB - Acute hemodynamic and humoral effects of synthesized alpha human atrial natriuretic polypeptide (alpha-hANP, 0.025 microgram/kg/min for 40 min) on 6 patients with severe congestive heart failure were assessed. Plasma alpha-hANP concentration was high in patients and increased further (from 463 +/- 360 to 1,282 +/- 670 pg/ml, mean +/- SD, p less than 0.01) following alpha-hANP infusion, but plasma norepinephrine (1,030 +/- 865 to 971 +/- 785 pg/ml) was not changed. Increases in urine output (1.0 +/- 0.8 to 2.6 +/- 2.3 ml/min) and Na+ excretion rate (87 +/- 89 to 257 +/- 211 mEq/min/m2) were statistically insignificant. A significant reduction was induced in mean aortic pressure (99 +/ 25 to 96 +/- 26 mmHg, p less than 0.05), mean right atrial pressure (11 +/- 9 to 7 +/- 8 mmHg, p less than 0.01), mean pulmonary arterial pressure (39 +/- 13 to 33 +/- 12 mmHg, p less than 0.05) and mean pulmonary capillary wedge pressure (27 +/- 8 to 20 +/- 7 mmHg, p less than 0.01). Heart rate, cardiac index, systemic vascular resistance and pulmonary vascular resistance were not altered. In conclusion, alpha-hANP induced decreases in left ventricular filling pressure and rightside heart pressure which were attributed to venodilatation rather than natriuresis in patients with congestive heart failure. Preserved cardiac output with decreased preload suggested that alpha-hANP improved cardiac function. PMID- 2969429 TI - Beneficial effects of coenzyme Q10 on impaired left ventricular performance in streptozotocin diabetic rats. AB - Chronic effects of coenzyme Q10 (CoQ: 30 mg/kg/day for 12 weeks) on cardiac performance in streptozotocin (45 mg/kg) induced diabetic rats were examined. Cardiac performance was assessed using the isolated retrograde perfused isovolumically contracting heart model. Compared to age matched nondiabetic rats, decreases occurred in myocardial CoQ (25.8 +/- 3.3 vs. 31.9 +/- 2.7 micrograms/ml), AMP (0.9 +/- 0.7 vs. 2.0 +/- 0.4 micrograms/mg), Emax (37 +/- 14 vs. 80 +/- 38 mmHg/microliter/g), an index of myocardial contractility, and LV diastolic chamber stiffness constant k (0.68 +/- 0.13 vs. 1.31 +/- 0.59 g/microliter) in diabetic rats. Normalized left ventricular weight (2.97 +/- 0.23 vs. 2.51 +/- 0.21 mg/g) and volume (1.53 +/- 0.34 vs. 0.89 +/- 0.53 microliter/g) and time constant of left ventricular pressure fall, T (32.0 +/- 8.0 vs. 19.7 +/- 2.6 ms) increased in diabetes. In diabetic rats taking CoQ, myocardial CoQ (28.5 +/- 3.2 micrograms/ml) and AMP (2.1 +/- 1.7 micrograms/mg) were the same as control, and T (23.5 +/- 7.4 ms) was significantly shortened (mean +/- SD, p less than 0.05, p less than 0.01). To compensate for depressed myocardial contractility and relaxation, LV dilatation and increased LV mass occurred in diabetic rats. Exogenous CoQ increased myocardial CoQ content and improved myocardial relaxation in diabetic rats. PMID- 2969430 TI - [A clinical experience with percutaneous transluminal angioplasty of bilateral stenotic renal arteries due to fibromuscular dysplasia]. PMID- 2969431 TI - [The skin-adipose tissue flap on the rectus abdominis muscle in reconstructive surgery of the breast]. PMID- 2969432 TI - Influence of hyperglycemia on Ca2+-Mg2+ -ATPase of red blood cells from diabetic patients. AB - The influence of hyperglycemia on calmodulin-stimulated Ca2+-Mg2+-ATPase activity was studied in 20 diabetic subjects. A clear inverse relationship between short term hyperglycemia as well as chronic hyperglycemia and calmodulin-stimulated Ca2+-Mg2+-ATPase activity could be demonstrated. Such a relationship could not be shown for basal enzyme activity. PMID- 2969433 TI - Serum lipoprotein Lp(a) concentrations are not influenced by an HMG CoA reductase inhibitor. PMID- 2969434 TI - Inhibitory effects of danazol and medroxyprogesterone acetate on [3H]thymidine incorporation in human endometrial cancer cells. AB - Effects of medroxyprogesterone acetate (MPA) and danazol (1 nM-10 microM) on cultured cancer cells from human endometrial adenocarcinomas obtained by hysterectomy were simultaneously investigated. Of twenty-four endometrial adenocarcinomas examined, five tumors were successfully maintained in primary cell culture. The addition of MPA as well as danazol in culture of cells from the five tumors resulted in a significant inhibition of [3H]thymidine incorporation in cancer cells from three tumors having progesterone receptors (PR). The minimum effective concentrations of MPA and danazol for the inhibition of [3H]thymidine incorporation were found to be 10 and 100 nM, respectively. The difference in effective concentration could be explained by a higher affinity of MPA to PR than that of danazol in cancer cells. On the other hand, neither danazol nor MPA affected [3H]thymidine incorporation in cultured cells from the remaining two tumors, in which PR was absent in one but present in the other. These findings, together with our previous findings that danazol inhibited the growth of a human endometrial cancer cell line with PR, suggest that a growth-inhibitory effect of danazol on human endometrial cancer cells is mediated through PR in the cells. PMID- 2969435 TI - Aminoglutethimide and ketoconazole: historical perspectives and future prospects. AB - Aminoglutethimide and ketoconazole, although originally developed as an anticonvulsant and antifungal agent respectively, have both been used to suppress steroid biosynthesis in patients with hormone-sensitive cancer. Aminoglutethimide inhibits several enzymes involved in the synthesis of corticosteroids as well as the aromatase enzyme which converts androgens to oestrogens. About one third of patients with breast cancer show objective improvement with aminoglutethimide, and it may also be of use in the treatment of adrenal carcinoma. However, its toxicity, and the need for concomitant cortisol replacement, severely limit its usefulness. Ketoconazole also inhibits several steroidogenic enzymes, notably C17,20-lyase, and has been used to treat carcinoma of the prostate. Again however, its toxicity and limited efficacy limit its value, although it may be useful in the treatment of certain endocrine conditions such as precocious puberty. Several aromatase inhibitors similar in structure to aminoglutethimide have been developed in an attempt to create more selective and efficient inhibitors. Some of these compounds have been tested in animals but none have as yet been subjected to clinical trials. Attempts to produce imidazole inhibitors of steroidogenesis are less advanced, although one compound (CGS 16949A) has been reported to be a more selective and potent aromatase inhibitor than aminoglutethimide. Selective and effective compounds could be of great value in the treatment of hormone-sensitive carcinoma. PMID- 2969436 TI - Early and late results of coronary artery bypass after failed angioplasty. Actuarial analysis of late cardiac events and comparison with initially successful angioplasty. AB - We reviewed the results of early (less than 24 hours) coronary artery bypass after unsuccessful percutaneous coronary artery angioplasty in 146 patients treated between October 1979 and July 1986. Overall operative mortality was 2.7%, and risk was significantly increased among patients with hemodynamic instability and new occlusion or further narrowing of the dilated vessel (3.8 versus 0%, p less than 0.05). Actuarial analysis was used to compute the rates of cardiac events during the follow-up interval, and event rates were also estimated in a comparison group of 776 patients who had successful first-time PTCA during the same time period. At a follow-up interval of 5 years, the cumulative risks of recurrence of angina and need for an additional procedure (bypass or angioplasty) were significantly (p less than 0.05) lower for patients who had undergone bypass than for those who had successful angioplasty (angina 21% versus 56%, PTCA 2% versus 21%, CAB 6% versus 16%). Cumulative risks of myocardial infarction and death were 4% versus 9% and 6% versus 9% in the two groups. The differences between late outcomes in the bypass and angioplasty groups persisted when patients were stratified into cohorts with single-vessel and multivessel disease, and the highest late event rate occurred in patients in the angioplasty group who had incomplete revascularization. The difference in late events after bypass or angioplasty was greatest during the first year. These late data should be considered when the mode of revascularization (bypass or angioplasty) is selected for symptomatic patients, especially those with multivessel disease. PMID- 2969437 TI - Emergency operation after failed angioplasty. AB - A group of patients with failed angioplasty who then required emergency coronary bypass was compared with a historically matched group of patients who had had elective bypass grafting. The two groups were well matched in age, sex, ejection fraction, and New York Heart Association classification and in the incidence of diabetes and hypertension. Significant differences were found in the prevalence of mortality (12% versus 1.5%), hemorrhage (28% versus 13%), cardiac tamponade (10.5% versus 1.5%), myocardial infarction (28% versus 9%), and length of hospital stay (15.3 days versus 13.4 days). Cardiogenic shock carries the worst prognosis; four of the five patients with this condition died. Because emergency operation after failed angioplasty carries with it significant postoperative morbidity and mortality, this procedure cannot be considered equivalent to elective coronary bypass grafting. PMID- 2969438 TI - Atrial natriuretic factor: response to cardiac operation. AB - Patients who undergo cardiopulmonary bypass experience derangements of complement activation, prostaglandin metabolism, and catecholamine secretion, which have all been posited to explain postoperative fluid retention and paroxysmal hypertension. Atrial natriuretic factor, a hormonal peptide released by the cardiac atria, regulates vascular volume by increasing sodium excretion and decreasing vasomotor tone and catecholamine synthesis. We examined its possible response to cardiopulmonary bypass in 23 patients (18 having cardiopulmonary bypass and, as a control group, five having thoracotomy) who underwent serial blood sampling to measure plasma atrial natriuretic factor levels before, during, and after operation. Thoracotomy alone had no effect on atrial natriuretic factor levels before, during, and after operation. Patients with cardiac valve lesions who had a higher incidence of arrhythmias and congestive heart failure showed elevated preoperative atrial natriuretic factor values (p less than 0.05). Normally, atrial natriuretic factor release is directly related to atrial filling pressure, but all patients having cardiopulmonary bypass revealed a paradoxical rise of atrial natriuretic factor during cardiopulmonary bypass (p less than 0.01) and a lack of correlation between atrial filling pressure and atrial natriuretic factor secretion in the early postoperative period. The atrial natriuretic factor response to pulmonary wedge pressure began to normalize 24 hours after operation. The unique reaction of this recently discovered cardiac hormone to cardiopulmonary bypass suggests its possible role in the pathophysiologic response to cardiac operation. PMID- 2969439 TI - Immune defects and therapeutic approaches to immune reconstruction in AIDS/ARC. PMID- 2969441 TI - The pathological association between Down syndrome and Alzheimer disease. AB - The neuropathology of Down syndrome (DS) at middle age is compared with that of Alzheimer disease (AD) at that age, through a review of the published literature and from the author's personal observations on brains from a series of patients of different ages with DS. It is noted that the pathological changes of DS at middle age (i.e. the form and distribution of senile plaques and neurofibrillary tangles, the pattern of involvement (atrophy) of neuronal systems) are qualitatively the same as those of AD at that age, though quantitative differences do occur and these may relate to biological or sociological variations inherent to the two parent populations. It is concluded that in pathological terms patients with DS at middle age do indeed have AD. Some ways in which a study of patients with DS can give insight into the nature and development of the pathological changes of AD are put forward and discussed. PMID- 2969440 TI - Lymphocyte proliferative response to PHA and anti-CD3/Ti monoclonal antibodies, T cell surface marker expression, and serum IL-2 receptor levels as biomarkers of age and health. AB - Alteration of T cell surface marker expression with a decrease of CD3 positive cells relative to the number of CD4 and CD8 positive cells, diminished in vitro proliferative response to mitogenic stimuli like PHA and antibodies to the CD3/Ti complex, and increase in serum IL-2 receptor levels, are among the changes in immunologic parameters that have been associated with advanced age. To distinguish between effects of the primary aging process and diseases of aging not known to be directly related to immune function, we investigated these variables in two well characterized populations of elderly donors (greater than 70 years) and a young adult control group (less than 35 years). The first group of older donors reported no evidence of significant chronic or recent acute illness and saw a physician only for routine medical care. The second group was randomly selected from individuals seen in a geriatric medicine clinic for diagnoses that included osteoarthritis and cardiopulmonary disorders. Altered surface marker expression and increased serum IL-2 receptor levels were seen only in the second group. On the other hand, lymphocyte proliferative responses to PHA, Leu 4 (anti-CD3) and a monoclonal antibody to the beta-chain of the T cell antigen receptor (WT31) were significantly decreased in both populations. Because we would expect primary aging to affect even extremely fit individuals of advanced age, we conclude that decrease in T cell proliferative response may represent a biomarker of primary aging in man. The alteration in surface marker expression and increased IL-2R levels in serum appear to be effects secondary to non-immunologic disease rather than aging. PMID- 2969442 TI - [Circulating immune complexes in acute meningococcal disease. Absence of a pathogenic role]. PMID- 2969443 TI - Cocaine influences beta-endorphin levels and release. AB - Immunoreactive beta-endorphin (IR-BE) was measured in the plasma, anterior pituitary (AP), neurointermediate lobe of the pituitary (NIL) and hypothalamus of male rats treated chronically (once daily for ten days) with cocaine. Cocaine produced a consistent elevation in the concentration of IR-BE in the plasma, the AP and the NIL at doses of 2.5 - 20 mg/kg/ip. The release of IR-BE from the AP and the NIL was determined in vitro and was found to be increased by treatment with cocaine. Chronic administration of cocaine did not affect the concentration of IR-BE in the hypothalamus. Chromatographic analysis revealed that cocaine produced a slight decrease in the amount of beta-endorphin relative to beta lipotropin in the AP. Beta-endorphin was the major form of IR-BE released by the AP and the sole constituent and secretory product of the NIL. These data indicate that chronic administration of cocaine stimulates the endogenous opiate system, elevating the levels of IR-BE in the pituitary and promoting beta-endorphin release. PMID- 2969444 TI - Intrarenal localization of degradation of atrial natriuretic peptide in isolated glomeruli and cortical nephron segments. AB - Using isolated glomeruli and nephron segments obtained from collagenase treated rabbit kidneys, we examined the in vitro degradation of alpha-human atrial natriuretic polypeptide (alpha-hANP). The ANP-degrading activity was measured by the amount of immunoreactive ANP remaining after incubation of about 50 fmoles alpha-hANP with each tissue preparation for 7.5 min. The sequence of degrading activity among isolated nephron segments was as follows: proximal straight tubule greater than proximal convoluted tubule greater than cortical collecting tubule greater than distal convoluted tubule greater than cortical thick ascending limb. A single glomerulus exhibited the degrading activity which was comparable to approximately 50% of the activity of 1 mm proximal convoluted tubule. Phosphoramidon, an inhibitor of endopeptidase, prevented the degradation of ANP in proximal convoluted tubule and glomerulus by 68% and 89%, respectively, but not in cortical thick ascending limb and cortical collecting tubule. From these results, we conclude that the degradation of ANP by endopeptidase occurs mainly in the proximal tubule and glomerulus. PMID- 2969445 TI - Vox Populi and Baby Doe. PMID- 2969446 TI - Lipid metabolic changes during hormonal treatment of endometriosis. AB - Three groups of patients with pelvic endometriosis were treated with medroxyprogesterone acetate (MPA) 50 mg/day (n = 10), lynestrenol (LYN) 10 mg/day (n = 25) and danazol (DAN) 600 mg/day (n = 25) respectively. Total cholesterol, triglycerides, alpha-lipoprotein, prebetalipoprotein, beta-lipoprotein cholesterol, high-density (HDL) and low-density (LDL) lipoprotein and apolipoprotein A1 and B concentrations were determined before treatment and after 3 and 6 mth of therapy. Whereas lipid, lipoprotein and apolipoprotein levels did not change in the MPA group, the patients on LYN and in particular those on DNA showed marked changes in lipoportein patterns. Alpha-lipoprotein, HDL and apolipoprotein A1 levels fell, while beta-lipoprotein, LDL and apolipoprotein concentrations rose, these changes being statistically significant. No alterations were seen in the serum levels of cholesterol, triglycerides or prebetalipoprotein cholesterol. In view of a possible relationship between high LDL and low HDL levels and a risk of accelerated coronary arteriosclerosis in women it was concluded that progestogen-induced alterations in lipoprotein patterns should be avoided as far as long-term treatment is concerned and where additional risk factors are present. PMID- 2969447 TI - Modeling AIDS and its treatment with immunostimulation. AB - A mathematical model of AIDS and its response to treatment with immunostimulation therapy is introduced. It correctly predicted the response of an AIDS patient to hyperimmunization with inactivated polio vaccine for a period of four months, indicating that this benign modality will offset the clinical effects of HIV-1 infection for almost a year. PMID- 2969448 TI - The dopaminergic D receptor: another example of reductive activation? AB - We have previously suggested that several intercellular messengers activate their receptors via reductive activation. Adenylate cyclase activation involves exposure of a sulfhydryl group. The dopamine D1 receptor activates this enzyme. Because sulfhydryl exposure could be secondary to reduction of a disulfide group we evaluated dopaminergic D1 agonists and antagonists as reducing agents. The agonists were found to be reducing agents and the antagonists were inactive. These results are consistent with the concept that dopaminergic D1 agonists activate adenylate cyclase via reductive activation. PMID- 2969449 TI - Cyclical neutropenia and retinoid therapy with isotretinoin. PMID- 2969450 TI - [Effect of human atrial natriuretic peptide on insulin levels and insulin secretion]. PMID- 2969451 TI - Identification and characterization of a Mg2+-dependent and an independent Ca+2 ATPase in microsomal membranes of rat testis. AB - Rat testicular microsomal membrane fraction contains both Mg+2-dependent and Mg+2 independent Ca+2-ATPase activity. The latter activity is about two times higher than the former. Calcium ion required for maximum activation of Mg+2-independent Ca+2-ATPase in 3.0 mM, whereas for the dependent one it is 2.5 mM. Both the enzymes are resistant to cold shock upto seven days. Histidine and imidazole buffers are found to be the most suitable for dependent and independent enzyme activities, respectively. The pH optima for dependent one is 7.5, whereas for the independent one it is 8.5. Temperature optima for the former is 37 degrees C and for latter one it is 40 degrees C. Among all the nucleotides tested, ATP is found to be the best substrate for both the enzymes. The optimum concentration of ATP for dependent and independent enzyme activities are 3.0 mM and 1.5 mM respectively. Divalent metal ions like Zn+2, Ba+2 and Mn+2 have been found to inhibit Mg+2-dependent Ca+2-ATPase activity whereas Mg+2-independent Ca+2-ATPase activity is inhibited by the divalent ions except zinc which is found to stimulate the enzyme activity. Both the enzymes are inhibited by vanadate, EDTA and EGTA. I50, for vanadate is 0.05 and 0.125 mM for dependent and independent activities, respectively. Sulfhydryl groups modifying agents e.g., NEM, DTNB and chlorpromazine are found to affect the enzyme activities in different ways. Thus NEM and chlorpromazine are found to inhibit and DTNB stimulate the enzyme activities in both the cases. PMID- 2969452 TI - Thyroid hormone increases rat atrial natriuretic peptide messenger ribonucleic acid accumulation in vivo and in vitro. AB - Thyroid hormone has a number of effects on cardiovascular and renal function which are shared by the atrial natriuretic peptide (ANP). We attempted to demonstrate a relationship between the two by studying the effects of thyroid hormone on the expression of the ANP gene and the secretion of its encoded protein. Thyroid hormone, when given to thyroidectomized rats, increased plasma ANP levels by approximately 2-fold in both watered and dehydrated animals. Cardiac ANP mRNA in dehydrated animals fell to 25% of that in the water-replete controls. T4 increased cardiac ANP mRNA 3-fold in dehydrated animals, but failed to alter ANP mRNA in those animals allowed free access to water. The effect of thyroid hormone appeared to take place, at least in part, at the level of the ANP synthesizing cardiocyte. T3, at concentrations ranging from 10(-10)-10(-8) M, increased ANP mRNA levels a maximum of 2-fold in primary cultures of neonatal cardiocytes. Both basal and T3-stimulated ANP transcripts appeared to be identical to their counterparts in the adult atria, as assessed by blot hybridization and S1 nuclease analysis. T3 (10(-8) M) also effected a 2-fold increase in media ANP immunoreactivity. These data indicate that thyroid hormone increases the secretion and genetic expression of ANP in vivo and in vitro and suggests a role for the peptide as a mediator of at least some thyroid hormone effects in the cardiovascular system. PMID- 2969453 TI - Cloning of a complementary deoxyribonucleic acid coding for human thyroxine binding globulin (TBG): existence of two TBG messenger ribonucleic acid species possessing different 3'-untranslated regions. AB - An adult human liver cDNA library constructed in expression vector, bacteriophage lambda gt11, was screened with polyclonal antibody directed against human T4 binding globulin (TBG). TBG cDNA cloned in the present study was 944 nucleotides in length. It contained approximately 70% of the coding region and complete 3' untranslated region. When the sequence was compared with that of TBG cDNA recently cloned by I. L. Flint, T. J. Bailey, T. A., Gustafson, B. E. Markham, and E. Morkin, the 3'-untranslated region of our cDNA was 231 nucleotides shorter than their cDNA. These results indicated that two TBG mRNAs with different length of 3'-untranslated regions may exist in human liver. Indeed, Northern blot analysis revealed that two TBG mRNAs differing in the length approximately 200 base pairs were present in normal human liver as well as in human hepatoma cell line (HepG2). It was demonstrated that this size difference was due to the length of 3'-untranslated region by hybridization with a probe specific to the longer 3' end. Together with the sequence data, it was suggested that these two TBG mRNA species may be produced by alternative processing and polyadenylation at two different sites. PMID- 2969454 TI - Molecular cloning and temporal expression during pregnancy of the messenger ribonucleic acid encoding uteroferrin, a progesterone-induced uterine secretory protein. AB - A lambda gt11 expression library containing cDNA inserts prepared from porcine endometrial mRNA was immunologically screened by using an antiserum developed against porcine uteroferrin (Uf), a glycoprotein that has been strongly implicated in transplacental iron transport in the pregnant pig. Antibody reactive clones (lambda 4a3, 13.1, and 2.2) were isolated after screening 1.5 x 10(5) recombinant phages. Clones 4a3 and 13.1 expressed Uf antigenic determinants in beta-galactosidase fusion proteins and specifically selected antibody which reacted with Uf in immunoblots prepared from uterine cytosolic extracts. In addition, all three cDNA clones collectively contained DNA sequences that encoded an 85-amino acid peptide which corresponded to a region within the carboxyterminal portion of the Uf protein. Northern blot hybridization of these cDNAs to RNAs extracted from whole uterine tissue of pregnant pigs revealed a single uterine poly(A)+ RNA of approximately 1.7 kilobases in length, which was not found in liver and mammary tissue RNAs. The concentration of the Uf mRNA changed in a temporal fashion during pregnancy in a manner that was distinct from that of the progesterone receptor mRNAs. Highest levels of Uf mRNA were found at mid and late pregnancy (days 45-110) and were about 50-fold greater than at day 30 of pregnancy. By contrast, RIA analysis of the uterine tissue extracts showed that maximum amounts of Uf were present at day 60 and then declined sharply. Thus the pattern of Uf mRNA present in the uterus did not parallel the amount of Uf polypeptide that could be recovered from the tissue. The tissue specific and temporal regulation of Uf gene expression emphasizes that the protein plays an important role in uterine activity and/or fetal development. PMID- 2969455 TI - Cloning of the gene encoding a 50 kilodalton potential surface antigen of Schistosoma mansoni. AB - A cDNA library was constructed from the mRNA of adult worms of Schistosoma mansoni, in the expression vector lambda gt11. This library was screened with a pool of sera raised against either soluble egg antigens or purified schistosomulum tegumental membranes. An antiserum raised against the fusion protein of one clone immunoprecipitated a 45 kDa polypeptide from the in vitro translation products of adult worm mRNA and recognised a 50 kDa antigen in homogenates of adult worms. This serum gave positive fluorescence of the surface of schistosomula in indirect immunofluorescence assays and was able to mediate killing of schistosomula by human eosinophils in vitro, suggesting that this clone contained part of a gene encoding a surface antigen. PMID- 2969456 TI - Atrial natriuretic factor in the Landry-Guillain-Barre syndrome. PMID- 2969457 TI - Behavioral significance of opioid peptides in relation to hippocampal function. PMID- 2969458 TI - Opioids in the hippocampus: progress obtained from in vivo electrophysiological analyses. PMID- 2969459 TI - Cell-cell interactions in the antibody response. PMID- 2969460 TI - Can B cells turn on virgin T cells? AB - The first event in the initiation of an immune response is the capture and presentation of antigen to T cells. Such presentation involves two distinct steps: (1) display of the antigen, which requires uptake, processing and re expression of the antigen in association with MHC molecules on the presenting cell surface; and (2) triggering, in which the presenting cell provides signals leading to the activation of the responding T cell. Two sorts of cells can capture antigens, the 'professional' antigen-presenting cells (APCs) such as dendritic cells and macrophages, and the B cells. Both types of cells can display antigens and the APCs are known to be able to trigger resting T cells. But despite in vitro evidence that certain B-cell types can reactivate previously activated T cells, it is not yet clear whether a B cell can initiate an immune response by providing the signals necessary to activate a resting T cell. We reasoned that resting B cells should not have this capacity because of the problems this would present with tolerance to self idiotypes. By exploiting the unique properties of the avian haematopoietic system, we have examined the presenting capacity of B cells in vivo and found that resting B cells are indeed unable to activate resting T cells. PMID- 2969462 TI - A comment on Dr. Page's article. PMID- 2969461 TI - A gene induced by the plant hormone abscisic acid in response to water stress encodes a glycine-rich protein. PMID- 2969463 TI - [Detachable balloon treatment of vertebral fusiform aneurysms]. AB - Percutaneous embolization using a detachable balloon in six cases of vertebral fusiform aneurysm is reported. Location of these aneurysms were as follows; 5 aneurysms in the distal vertebral artery (2 proximal to the posterior inferior cerebellar artery; PICA, 2 distal to the PICA and 1 where the PICA was undetectable), and one aneurysm in the extracranial vertebral artery. Three cases suffered from subarachnoid hemorrhage (dissecting aneurysms) and other cases showed lower cranial nerve palsy (giant aneurysm) or posterior fossa ischemia. Most of the cases were treated immediately after conventional diagnostic angiography, especially those cases with subarachnoid hemorrhage (within 4 days after the last attack). Under local anesthesia, a catheter was positioned in the affected vertebral artery either trans-femorally or trans-axillary. The detachable balloon was placed proximal to the aneurysm and test occlusion was performed for 15-20 minutes (the tolerance test or balloon Matas' test). After it was confirmed that there had been no neurological deterioration, the vertebral artery was permanently occluded by detachment of the balloon. The procedure was successful in all cases. Thrombosis of aneurysms was confirmed either by follow up angiography or by autopsy. However, two cases who were grade V (Hunt & Kosnik) at admission died of severe vasospasm. PICA was preserved through either ipsilateral or contralateral collaterals. Detachable balloon occlusion of the vertebral artery for fusiform aneurysm is thought to be less invasive, more indicative and a safer procedure compared to conventional surgical proximal occlusion therapy. PMID- 2969464 TI - 'Subcortical dementia': the neuropsychological evidence. PMID- 2969465 TI - [Postcoital contraception using a desogestrel-ethinyl estradiol combination]. PMID- 2969466 TI - [A cyproterone acetate-ethinyl estradiol combination in the treatment of acne. Data of a survey of gynecologists]. PMID- 2969467 TI - Rapid conversion of high into low striatal D2-dopamine receptor agonist binding states after an acute physiological dose of 17 beta-estradiol. AB - Ovariectomized female rats injected with 17 beta-estradiol (100 ng, s.c.) showed, as previously observed, an increase of the dopamine (DA) metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) with no change of DA concentrations in the striatum. This increase was observed as soon as 15 min following the injection while plasma estradiol reached a peak of 78 pg/ml after 5 min and was significantly elevated until 45 min to ultimately return to control values at 60 min. We observed no significant change of the inhibition constants of high- and low-affinity D2 DA agonist binding sites and of the sum of high + low agonist DAergic agonist binding densities as detected by apomorphine competition of [3H]spiperone binding. By contrast, a significant conversion of high into low agonist affinity binding states was seen at 15 min (38.6% of conversion, P less than 0.05) and 30 min (40.0% of conversion, P less than 0.01) after the acute physiological steroid injection. Thus, very small doses of estradiol were able to rapidly increase DA turnover and modulate the striatal agonist affinity states of the D2 DA receptor. This effect of estradiol is probably non-genomic, presynaptic and may involve a membrane effect at the DA autoreceptor level. PMID- 2969468 TI - Dopamine D1- and D2-receptor interaction in turning behaviour induced by dopamine agonists in 6-hydroxydopamine-lesioned rats. AB - The selective dopamine D2-antagonist sulpiride potentiated contralateral circling behaviour induced by the D1-agonist CY 208-243 in rats with unilateral lesions of substantia nigra, but reduced the effects of the selective D2-agonist bromocriptine. Similarly, the D1-antagonist SCH 23390 tended to increase the effects of bromocriptine but markedly inhibited CY 208-243 induced turning. The mixed D1/D2-antagonist fluphenazine was effective in reducing circling behaviour induced by either agonist, whereas pimozide (D1/D2) inhibited only the actions of bromocriptine. These results indicate that the actions of CY 208-243 and bromocriptine are mediated via distinct but interacting receptor subtypes. PMID- 2969469 TI - The existence of low concentrations of atrial natriuretic peptide (ANP) in canine cerebrospinal fluid which does not correlate with plasma ANP levels. AB - Atrial natriuretic peptide (ANP) concentrations in the cerebrospinal fluid (CSF) and plasma of canine were 2.1 +/- 1.1 pg/ml (mean +/- S.D.) and 53.1 +/- 21.1 (n = 20), respectively. The regression coefficient between these concentrations was 0.0045 (P = n.s.). The ANP concentration in the CSF did not change even after the plasma ANP concentration was altered following the change of left atrial pressure, as in 4 cases of an experimental aortic regurgitation. Thus, ANP concentration in the CSF is not influenced by ANP concentrations in the plasma at least under our condition. Gel permeation chromatography revealed a single form of ANP in the position of authentic alpha-ANP in canine CSF, while a high molecular weight ANP peak was observed as well as alpha-ANP in the plasma. PMID- 2969470 TI - Hepatitis B immunisation. PMID- 2969471 TI - [Heavy metal burden of children in the Ruhr area]. PMID- 2969472 TI - [AIDS from the ethical and theologic perspective]. PMID- 2969473 TI - [Legislation concerning standardization of public health--studies of anamnesis and reflections on prognosis]. PMID- 2969474 TI - [A model for the community management of psychiatric patients--Bregenz social psychiatry]. PMID- 2969475 TI - [Prevention from the nutrition medicine viewpoint]. PMID- 2969476 TI - [The status of environmental and public health engineers in public health and environmental bureaus]. PMID- 2969477 TI - [Causal relation of pathogen and infection. I]. PMID- 2969478 TI - [Probability of detecting Salmonellas in waste water, clearing sludge and drainage samples]. PMID- 2969479 TI - [Bacteriologic studies of a sewage plant equipped with a biofiltration stage in Mannheim. 1. Determination of total pathogen numbers, gram-negative rod bacteria (except Salmonella), Enterococci and Staphylococcus aureus]. PMID- 2969480 TI - [Options for medical consultation in health insurance by specially appointed physicians in inpatient treatment]. PMID- 2969481 TI - [New results in the Heilbronn "Performance and cost evaluation trial in legal health insurance"]. PMID- 2969482 TI - [Preventive vaccination of school children in the Marburg-Biedenkopf area]. PMID- 2969483 TI - [Development of medical laboratory diagnosis in social medicine offices]. PMID- 2969484 TI - [Effect of cigarette smoke on tear secretion and tear film stability]. PMID- 2969485 TI - [Biotropic effects caused by atmospheric electrical factors in enclosed spaces]. PMID- 2969486 TI - [Neuropediatric aspects in learning disability]. PMID- 2969487 TI - [Significance of social work for public health administration]. PMID- 2969488 TI - [Incidence and trends of suicide attempts in West Germany--a methodological study]. PMID- 2969489 TI - [Significance of Listeria monocytogenes in foods]. PMID- 2969490 TI - [Causal relations between pathogen and infection. II]. PMID- 2969491 TI - [Bacteriological studies of a sewage plant of the city of Mannheim equipped with a biofiltration stage. II. Determination of Salmonella serotypes]. PMID- 2969492 TI - [Founding and development of social pedagogic work in the public health administration]. PMID- 2969493 TI - [Shadow projection--a factor in the HIV/AIDS problem]. PMID- 2969494 TI - A diagnosis of AIDS: understanding the psychosocial impact. AB - Acquired immunodeficiency syndrome (AIDS) was discovered in 1981. It is now estimated that 1.5 million persons have become infected and that, by the year 1991, there will be 270,000 cases of the disease and 179,000 associated deaths. An extraordinary aspect of the AIDS epidemic is the high level of fear manifested by large numbers of people, in a manner disproportionate to the objective threat. AIDS has been referred to in the press as the greatest public health problem in America today. While dental journals are replete with studies enumerating the oral manifestations, and appropriate infection control guidelines, the psychosocial aspects of AIDS have generally been confined to media reports. Appropriate dental management of patients at high risk for the development of AIDS/AIDS-related complex requires a complete understanding of the psychosocial environment confronting these persons. PMID- 2969495 TI - Human T-lymphocyte subpopulations in periapical lesions. AB - Both B-lymphocyte-and T-lymphocyte-mediated immunologic reactions have been implicated in the pathogenesis of human periapical lesions. The purpose of this study was to identify various subpopulations of T-lymphocytes in these lesions. Fifteen periapical lesions were obtained during periapical surgery. Each specimen was quick-frozen and stored in liquid nitrogen. Immunoperoxidase staining for the presence of T-lymphocyte subpopulation, was done with the use of the monoclonal antibody and Vecta-stain ABC/peroxidase kit. Cryostat tissue sections were incubated with 3% normal horse serum. Specific monoclonal antibody (Coulter T4, T8, T11) was placed on the section, washed, and then covered with biotinylated antibody to mouse IgG. After TRIS HC1 washing, the ABC/peroxidase solution was placed on the section and 3,3-diaminobenzidine was applied; the section was washed and then counterstained with 2% methyl green. After dehydration, sections were mounted and examined under the light microscope. The presence of lymphocytes was confirmed by the appearance of brown rings around cells with lymphocyte morphology. Fourteen lesions were diagnosed as granuloma and stained positively for pan T-lymphocytes (T11), T helper cells (T4), and T cytotoxic cells (T8). The remaining specimen, diagnosed as an apical scar, contained no T-lymphocytes. The presence of T-cells in periapical lesions indicates that cell-mediated reactions participate in the pathogenesis of these lesions. PMID- 2969496 TI - Chromosomal localization of the human genes for lipocortin I and lipocortin II. AB - The human genes which code for Lipocortin I and Lipocortin II, proteins that inhibit phospholipase A2 (PLA2) activity, have been regionally localized in the human genome by chromosomal in situ hybridization and segregation analysis in somatic cell hybrids using cDNA clones for Lipocortin I and II. Lipocortin I, the 35 kd substrate for the epidermal growth factor (EGF) receptor/kinase, maps to chromosome region 9q11- greater than q22. The Lipocortin II cDNA probe detects at least four independently segregating loci which map to human chromosome regions 4q21-q31.1, 9pter-q34 proximal to c-abl, 10q proximal to 10q24 and 15q21-q22 proximal to the 15q22 translocation breakpoint characteristic of acute promyelocytic leukemia (APL). Thus, Lipocortin I and one locus detected by Lipocortin II cDNA are syntenic on chromosome 9; one Lipocortin II locus is perhaps not far from the genes for EGF and IL-2 on 4q; and another of the Lipocortin II loci is on 15q, perhaps not far from the APL breakpoint. PMID- 2969497 TI - Spine x-ray abnormalities in idiopathic limb pain problems. Study notes possibility pain may be referred. AB - Five hundred and thirty-eight cases of spine (298) and limb (240) pain problems were coded for spine x-ray abnormality (congenital, scoliosis, straightening, and degenerative joint disease). Analysis reveals that the limb pain cases (97 upper and 143 lower) when compared with the spine pain cases (45 cervical and 253 lumbar) have fewer normal spine x-rays with a greater incidence of degenerative joint disease in the limb pain cases and a greater incidence of straightening in younger spine and limb pain cases. Excluding 107 disability cases increases these tendencies. A group of 191 women who have had x-rays every five years are studied for controls and show a much lower incidence of x-ray changes. These data support a diagnosis of referred pain from the spine as a significant possibility in the idiopathic limb pain problem case. PMID- 2969498 TI - Use of manipulative therapy for mechanical pain of spinal origin. AB - Manipulation of the spinal joints for the relief of pain is an ancient remedy that is in widespread use today. Clinical trials investigating the effectiveness of manipulative therapy are briefly reviewed, and some experimental evidence suggesting possible causes of spine pain is presented. The rationale for the use of spinal manipulative therapy within the context of R. A. McKenzie's diagnostic and therapeutic system is detailed and explicitly stated. Spinal mobilization and manipulation are presented as a logical treatment progression from the use of postural instruction and exercise therapy. Extensive description of technique is not given, but the clinical signs that indicate the need to use or avoid mobilization and manipulation in a particular direction are detailed. PMID- 2969499 TI - Monoclonal antibody-ricin conjugate cytotoxic to cells expressing the common acute lymphoblastic leukemia antigen (CALLA). AB - The monoclonal antibody PHM-6, which is specific for the common acute lymphoblastic leukemia antigen (CALLA), was conjugated to the plant toxin ricin. Binding of the PHM-6-ricin conjugate to cells via the ricin molecule was blocked by the presence of 100 mM lactose. The IC50 (concentration resulting in 50% inhibition) of the PHM-6-ricin conjugate for the CALLA-positive KM-3 cell line was 280-fold greater than for bone marrow stem cells, indicating the potential of this conjugate for immunological purging of autologous remission marrow. PMID- 2969500 TI - Screening the newborn for anatomic and metabolic defects. PMID- 2969501 TI - Communicating with parents of high risk infants. PMID- 2969503 TI - [Percutaneous transluminal atherectomy for iliac artery stenosis]. PMID- 2969502 TI - Comparison of attitudes of regular and adapted physical educators toward disabled individuals. AB - The Attitudes Toward Disabled Persons Scale (Form B) and a ranking survey of 10 disabling conditions were administered to 38 physical education teachers and 47 adapted physical education teachers. Both groups of teachers' attitudes toward the disabled were below the scale median. Both groups of teachers were more accepting of individuals with physical disabilities such as, amputee, harelip, and epilepsy, than individuals who were mentally retarded, cerebral palsied, and emotionally disturbed. PMID- 2969504 TI - [Radiation-induced expression of Ia antigens and lymphocyte infiltration in murine tumors]. PMID- 2969506 TI - The effect of in vitro and in vivo toluene exposure on rat erythrocyte and synaptosome membrane integral enzymes. AB - The mechanism of the anaesthetic effect of toluene on the central nervous system (CNS) was studied by using rat erythrocyte and synaptosome membranes as nerve cell models both in vitro and in vivo. The activities of the membrane-bound integral enzymes acetylcholinesterase (AChE), total adenosine triphosphatase (total ATPase) and magnesium-activated adenosine triphosphatase (Mg2+-ATPase) were determined. A short-term exposure to 2000 p.p.m. of toluene had an inhibitory effect on the enzyme activities studied. The degree of inhibition in erythrocyte membranes in vitro and in vivo, and in synaptosome membranes in vitro were in good correlation. In in vivo conditions, the synaptosome-bound enzymes were, however, significantly more inhibited by toluene, which indicates that membranes in vivo are even more vulnerable to the toxic effects of organic solvents than they are as isolated membranes in vitro. However, our results show that in vitro experiments can be used to predict the toxic nerve cell membrane effects of organic solvents. Toluene caused similar enzyme inhibitions both in neural cell membranes and in erythrocyte membranes. Thus, even peripheral non excitable cell membranes, like erythrocytes, can be used as nerve cell membrane models in studies on the mechanism of the anaesthesia caused by solvents. PMID- 2969505 TI - Purification and enzymological characterization of DNA-dependent ATPase IV from the Novikoff hepatoma. AB - DNA-dependent ATPase IV has been purified to near homogeneity from the Novikoff rat hepatoma. The enzyme is devoid of DNA polymerase, RNA polymerase, exonuclease, endonuclease, phosphomonoesterase, 3'- or 5'-phosphodiesterase, polynucleotide kinase, protein kinase, topoisomerase, helicase or DNA reannealing activities at a detection level of 10(-5) to 10(-7) relative to the ATPase activity. The enzyme is a monomer of Mr 110,000, has a sedimentation coefficient of 5.9 S, a Stokes radius of 40 A and a frictional coefficient of 1.32. In the presence of Mg2+ ion and a polynucleotide effector, ATPase IV hydrolyzes either ATP or dATP to the nucleoside diphosphate plus Pi. Other ribo- or deoxyribonucleoside triphosphates are not substrates. ATPase IV utilizes double stranded DNA and single-stranded DNA as effector; however, it does not utilize poly(dT). The Km for dsDNA or ssDNA is 2.2 microM (nucleotide). A variety of ATP analogues were found to be competitive inhibitors of ATPase IV. PMID- 2969507 TI - Neonatal treatment with monoamine uptake inhibitors alters later response in behavioural 'despair' test to beta and GABA-B receptor agonists. AB - The administration of monoamine uptake-inhibiting antidepressant drugs to rats during the early postnatal period was previously shown to lengthen the duration of subsequent immobility in Porsolt's swim test, hence suggesting increased behavioural 'despair' in these animals. Because the mechanism of the antidepressant action may be related to changes in the cerebral monoamine or gamma-aminobutyric acid (GABA) function, the present study was carried out to examine the response in the swim test to a beta-receptor agonist salbutamol, or to the GABA-B receptor agonists progabide and baclofen in rats treated with antidepressant drugs during the second and third postnatal week: either desipramine 5 mg/kg, nomifensine 10 mg/kg or zimeldine 25 mg/kg. When tested a month later i.e. at the age of two months these rats were immobile in water for a longer period than the controls. Salbutamol 10 mg/kg and progabide 100 mg/kg increased the immobility time in the control rats but neither drug affected the rats treated with desipramine, nomifensine or zimeldine. When the animals were 5 months of age, salbutamol 10 mg/kg and baclofen 10 mg/kg shortened the immobility time in the desipramine-treated rats. The control rats and those treated with zimeldine were not affected by the drugs. The results indicate that in the rats which were neonatally treated with antidepressants, the immobility time in water is lengthened in adulthood. Moreover, the response to beta-receptor and GABA-B receptor agonists is increased from the response observed in the control rats. PMID- 2969508 TI - Binding of toremifene to human serum proteins. AB - The in vitro protein binding of toremifene in human serum was measured by ultracentrifugation using 3H-toremifene together with unlabeled toremifene, 50, 500, and 5000 ng/ml. Of the total radioactivity 99.7 per cent was bound to the proteins independent of the concentration of the unlabeled drug. Binding of toremifene to different protein fractions was studied by adding 3H-toremifene and 500 ng/ml of cold toremifene to normal serum. The serum samples were exposed to agarose gel electrophoresis to fractionate different proteins. The radioactivity was localized using a position-sensitive proportional counter. After that the proteins were visualized by staining. Of the total protein bound radioactivity 92 per cent was bound to albumin, about 6 per cent to beta 1 globulin fraction and about 2 per cent to a fraction between albumin and alpha 1 globulins, part of this probably to alpha 1 acid glycoprotein. PMID- 2969509 TI - Fetal curarization for prenatal magnetic resonance imaging. AB - Fetal magnetic resonance (MR) imaging was performed at 33 weeks of gestation for investigation of a posterior fossa abnormality found at ultrasound screening. Fetal movements were abolished by vecuronium injected under ultrasound guidance into the umbilical vein. MR images showed atrophy of the left cerebellar lobe with cisternal dilatation. These were confirmed postnatally by CT scan. PMID- 2969510 TI - [Modern imaging in cardiology]. PMID- 2969511 TI - [Significance of anti-cytoskeleton antibodies in autoimmune diseases]. PMID- 2969512 TI - [Inoperable cancers of the upper respiratory and digestive tracts. Value of chemotherapy: 185 cases]. AB - A series of 185 squamous cell carcinomas of the head and neck was retrospectively analyzed. Induction chemotherapy was systematically administered. The overall tumour response rate was 38 per cent, and 39 tumours (22.4 per cent) became resectable after chemotherapy. The survival of complete responders was statistically higher than that of non or partial responders. Complete responders treated either by radiation therapy or surgery had similar survivals. Surgery improved the prognosis and reduced the rate of local and regional failures in non responders, including poor responders (less than 50 per cent). PMID- 2969513 TI - [Folate deficiency in medical intensive care: incidence, predictive factors and consequences. Results of a prospective study with 105 patients]. AB - Among 105 patients admitted to a medical intensive care unit over a five-month period, 53 per cent had a fall in serum folate concentration (less than 5 ng/ml), and 19 per cent had a concomitant fall in erythrocyte folate concentration (less than 170 ng/ml). Acute folate deficiency with severe haematological changes was observed in 2 patients. Apart from these 2 cases, the usual haematological parameters were not significantly different in patients with or without folate deficiency. On admission, there was a significant correlation between folate status, severity index and serum albumin and transferrin, all variables which reflect the patient's nutritional status. Patients with infection and fever, and those who have recently been operated upon are at a higher risk of folate deficiency. These findings suggest that patients admitted to a medical intensive care unit should receive an early supplement of folic acid. PMID- 2969514 TI - [Disorders of plasma trace elements in diabetes. Relation to blood glucose equilibrium]. AB - The influence of glycaemic control and diabetes characteristics on plasma concentrations of magnesium, zinc, copper, selenium, rubidium and bromine has been evaluated in 44 diabetics (30 insulin-dependent, 14 non insulin-dependent), and the results obtained were compared to those of 309 control subjects of the same mean age. Diabetics had reduced plasma magnesium concentrations (P less than 0.01) but normal erythrocyte magnesium levels. Plasma zinc and selenium concentrations were reduced, whereas those of copper were increased and those of bromine and rubidium were normal. Correlation between glycaemic control, evaluated by measurement of glycosylated haemoglobin levels, and each of the parameters studied was only demonstrated with magnesium in insulin-dependent diabetics (r = -0.561; P less than 0.02). No correlation was found with the other clinical or anthropometric characteristics of the diabetic patients studied. Diabetes seems to be associated with numerous abnormalities of plasma trace elements and magnesium, but the mechanism of these abnormalities has not yet been elucidated. A decrease in zinc and selenium concentrations and an increase in copper concentrations might be additional factors of atherogenicity. PMID- 2969515 TI - [Severe congenital laryngeal stridor. Endoscopic surgical treatment]. AB - Congenital laryngeal stridor or laryngomalacia is a congenital disease causing an usually shrill and solitary inspiratory noise, sometimes associated with disorders of deglutition and dyspnea when crying. Most often, the symptoms spontaneously disappear before the age of two. However, some cases are very severe, with permanent dyspnea, leading to tracheal intubation or tracheotomy. To avoid the risks of prolonged tracheotomy in infants, a new surgical technique has recently been described, beside epiglottectomy and hyomandibulopexy, now abandoned. This is simple endoscopic section and resection of the ary-epiglottic folds. Preliminary results in 15 patients seem to show the superiority of this so called epiglottoplasty technique. Except in one case with supraglottic oedema attributed to a major gastro-esophageal reflux, all patients recovered from their dyspnea with this procedure. Extubation usually was rapidly feasible and the post operative period was uneventful. The patients are kept in hospital for 2 to 5 days, and an antibiotic and anti-reflux treatment is recommended. This procedure is advocated as a treatment of choice of "laryngomalacia" with severe dyspnea. PMID- 2969516 TI - [Specific desensitization in 1988]. AB - Specific desensitization is one of the most widely used treatments in children and adolescents. It was introduced 65 years ago, but its scientific foundations have only been established during the last 10 years. At the beginning of this decade, the development of standardized allergens and improvements in the technique of administration inclined us to consider that desensitization was a promising technique. In 1988, however, these hopes must be mitigated for several reasons. Desensitization is effective when used appropriately and with a strict methodology, but the advent of potent allergens has increased its risks. Desensitization is now criticised, sometimes severely, on account of its efficacy/safety ratio and its duration. It seems that more than one-half of patients allergic to vespid venom can discontinue desensitization after 3 years, but the treatment lasts much longer in other allergies, and some patients sensitized to perennial allergens should be injected for life, which raises the problem of the cost of desensitization. At the moment, this method is regarded as indispensable to prevent allergy to Hymenoptera, useful in patients with moderate asthma due to allergy to house mites, and to be envisaged in some patients allergic to pollens or to cats. If desensitization has a future, it will reside in allergens at least equal in quality to those now available but safer, and also perhaps in techniques using monoclonal or anti-idiotype antibodies. PMID- 2969517 TI - [Cephalic duodenopancreatectomy with preservation of the pylorus]. AB - We report a technique of cephalic pancreatico-duodenectomy preserving the pylorus which differs from that initially devised by Longmire and Traverso. Restoration of digestive tract continuity consists successively of end-to-end duodeno-jejunal anastomosis, end-to-side choledoco-jejunal anastomosis and end-to-side pancreatico-jejunal anastomosis. The simple technique has the advantage of restoring a perfectly physiological circuit. It seems to produce satisfactory immediate and mid- or long-term results with improvements in the patients' comfort. PMID- 2969518 TI - [Rearrangement of the gene of the T-lymphocyte antigen receptor in mycosis fungoides: existence of 2 monoclonal populations?]. PMID- 2969519 TI - [Association of C2 complement fraction deficiency and diffuse atheroma in a young subject]. PMID- 2969520 TI - [Impact of contamination during the procurement of kidney transplants]. PMID- 2969521 TI - [Acute hepatitis and renal insufficiency associated with seroconversion to human immunodeficiency virus]. PMID- 2969522 TI - [Hypoprothrombinemia under cefotaxime during a vitamin K-deficient diet]. PMID- 2969523 TI - [A case of disseminated Penicillium marneffei penicilliosis in a patient with acquired immunodeficiency syndrome]. PMID- 2969524 TI - [A rare cause of meningeal hemorrhage: dissection of the extracranial internal carotid artery]. PMID- 2969525 TI - [Temporal status epilepticus disclosing infectious mononucleosis with encephalitis]. PMID- 2969527 TI - [Cardiac beriberi of fulminating course]. PMID- 2969526 TI - [The timing of angioplasty in acute myocardial infarction]. PMID- 2969528 TI - [Effects of smectite on colonic fermentations]. PMID- 2969529 TI - [Regressive painful thoraco-abdominal neuropathy in insulin-dependent diabetes]. PMID- 2969530 TI - [On the good use of cardiac pacemaker programming]. PMID- 2969531 TI - [Block of 21-hydroxylase of late manifestation. Clinical and biological aspects and therapeutic implications]. PMID- 2969532 TI - [Improvement of the prognosis of Basedow's disease by using high doses of carbimazole]. AB - The influence of synthetic antithyroid drug dosage on the course of Graves' disease was evaluated by comparing two groups of matched patients treated with carbimazole. Thirty-seven patients received a rapidly degressive treatment (60 mg/day initially rapidly reduced without replacement thyroid hormone therapy), and 36 patients received a prolonged treatment in high doses (60 mg daily for 6 months with progressive reduction of dosage over 5 months and replacement therapy). There was a highly significant difference in the actuarial curves of patients without relapse during 36 months: 82 per cent in patients with high doses, as against 38 per cent in patients with rapidly degressive treatment (log rank: chi 2 = 7.67, P less than 0.01). The anti-TSH receptor antibody titers decreased more rapidly in patients under prolonged treatment with high doses than in those with the rapidly degressive treatment. It is concluded that carbimazole in high doses is more immunosuppressive than in rapidly degressive doses and that it improves the prognosis of Graves' disease. PMID- 2969533 TI - [Photoplethysmography and laser Doppler velocimetry in the study of cutaneous vasomotility at high altitudes]. AB - During the second French scientific medical expedition to the Himalaya (ARPE, Annapurna IV, 1985), modifications of cutaneous circulation in hypoxia were studied. Two techniques were used for the first time to evaluate cutaneous vasoactivity under such conditions: laser Doppler velocimetry and photoplethysmography. Measurements were performed on the pulp of the third and fourth fingers of the right hand in 6 subjects native of sea-level countries: first at sea level, then in hypoxia after 15 days at 4,800 m before and after ingestion of a vasodilator drug (nicardipine 20 mg). Cutaneous circulation was explored under three thermal conditions: baseline (ambient temperature); after vasodilation obtained by immersion of the hand in hot water (40 degrees C), and after immersion of the hand in cold water (12 degrees C). In prolonged hypoxia a reduction in vasoconstriction at 12 degrees C was observed with both techniques. No significant change was observed at 40 degrees C. After dosing with nicardipine, the results differed according to the exploratory technique: a slight increase of the laser Doppler signal (NS) and a decrease of the photoplethysmographic signal were recorded. Acclimatization to cold and to hypoxia may concur to determine this response of cutaneous circulation. The action of vasodilating agents on cutaneous microcirculation in prolonged hypoxia is not clear and deserves further investigation. These drugs must be used with caution in the prevention and treatment of frostbite. PMID- 2969534 TI - [Surgery of the aortic arch in coronary patients. 400 surgically treated patients]. AB - Out of 400 consecutive patients who underwent aorto-iliac surgery 45 (11.25 per cent) were known to have coronary disease at the time of the operation. Pre operative coronary arteriography was carried out in the 11 patients (2.75 per cent) who had unstable or recently worsened angina. Aorto-coronary bypass was performed preventively in 7 patients (1.75 per cent of surgical patients). Mortality from preventive aorto-coronary bypass and secondary aorto-iliac surgery was nil. A study of post-operative mortality (2.75 per cent overall) showed that only one patient (0.25 per cent of surgical patients) died of myocardial infarction. The low rate of post-operative morbidity and mortality of coronary origin therefore does not incline to extend the indications for preventive aorto coronary bypass. However, it seems desirable to improve the indications for pre operative coronary arteriography by performing thallium 201 myocardial scintigraphy with dipyridamole infusion in all coronary patients about to undergo aorto-iliac surgery when no exercise electrocardiogram can be obtained because of intermittent claudication. This type of examination will probably broaden the indications for pre-operative coronary arteriography and preventive aorto coronary bypass, though presumably to a very small extent. Among these 400 patients who underwent aorto-iliac surgery and were all followed up for 2 to 12 years, only a few (0.25 per cent) benefited secondarily from coronary surgery, no doubt because the physiological age of that population was often too high. In fact, these patients were mainly at risk of cancer aggravated by both alcohol and smoking, for in this study, and rather in contrast with most publications, cancer was responsible for 44 per cent of secondary deaths, i.e. more than twice the percentage (20 per cent) of secondary deaths of cardiac origin. PMID- 2969535 TI - [Home thrombolysis for myocardial infarction. A multicenter study of the feasibility and evaluation of short-term prognosis]. AB - It has been proven since 1986 that in myocardial infarction the sooner thrombolysis is performed the better. Forty-four patients were selected to enter a double-blind randomized trial in which they received either an acylated plasminogen streptokinase activator complex or a placebo. The injections were given intravenously at home within the first 3 hours (within the first 2 hours in 26 of them), by doctors from Mobile care units. This home treatment in the acute phase made it possible to gain 75 minutes on average, and up to 90 minutes when it was performed by an anaesthetist trained in emergency management. No serious complication, such as haemorrhagic or allergic reaction, occurred, and arrhythmia was no more frequent in the treated group than in the placebo group. Home thrombolysis did not delay admission to a cardiology Intensive Care unit (66 min. versus 64 min). Mean coronary patency was 75 per cent, and up to 82 per cent, in patients treated within 2 hours of the first symptoms. There was no significant difference between areas of reperfused or not reperfused patients in relation to time (P less than 0.08). Diagnosis sensitivity was 100 per cent. Thus, home thrombolysis is feasible and safe when performed by trained emergency medical teams and when criteria for inclusion and exclusion are fulfilled. PMID- 2969536 TI - [Immunogenicity of a hepatitis B vaccine obtained by genetic recombination and containing products of S and pre-S2 genes]. AB - Immunization against hepatitis B by means of a vaccine obtained by genetic recombination on chinese hamster ovary cells was attempted in 32 adult subjects. The HBs antigen was purified from the culture supernatant and contained S and pre S2 genes products. Three 20 micrograms doses were injected intramuscularly at intervals of one month. Anti-HBs seroconversion levels and the geometrical mean of antibodies were slightly higher than those observed with plasma vaccines or genetically engineered yeast-derived vaccine. Antibodies directed against HBs appeared more rapidly and at a higher titre. Anti-pre S2 antibodies were detected after the third injection in 84 per cent of the subjects vaccinated. This recombinant hepatitis B vaccine prepared from chinese hamster ovary cells will probably be as effective as the first generation vaccines obtained by purifying the HBs antigen obtained from the blood of asymptomatic carriers. PMID- 2969538 TI - [Ventilation in prone position in acute respiratory distress syndrome of severe course. 3 cases]. PMID- 2969537 TI - [Vascular anastomoses of small caliber vessels. Comparison between continuous or interrupted sutures]. AB - Continuous sutures are generally used to perform anastomoses on vessels less than 2 mm in diameter, but some surgeons advocate interrupted sutures which they claim cause less stenosis and provide better vascular potency in the long term. Arteriovenous fistula for chronic haemodialysis is a good model of anastomosis on 1 to 2 mm wide vessels. We used this model to compare the 2 techniques in 2 randomized groups of 20 patients each. The type of suture (continuous or interrupted) was drawn by lot in non diabetic patients who were having their first fistula for chronic haemodialysis. All anastomoses were performed by end-to side implantation of a superficial vein onto the radial artery. Prolene 8/0 was used as suture material. The maximum follow-up period was 2 years. No significant difference in vascular potency could be demonstrated between the 3 groups. Thus, continuous sutures performed under suitable conditions can be used to perform with greater ease anastomoses which are as patent as those performed with interrupted sutures. PMID- 2969539 TI - [Inflammatory patches at the point of injection of calcium heparin reflecting eczema caused by heparin: first case of an unrecorded side effect of heparin therapy]. PMID- 2969540 TI - [Reduction of ventricular tachycardia by Valsalva's maneuver]. PMID- 2969541 TI - [Reconstruction of the tongue and mouth floor. Value of the free radial forearm transplant]. PMID- 2969542 TI - [Systemic Yersinia infection of cutaneous origin]. PMID- 2969543 TI - [Myasthenia caused by tiopronin during treatment of rheumatoid polyarthritis]. PMID- 2969544 TI - [Emergency screening for human immunodeficiency virus carriers among patients in irreversible coma as a potential source of transplantable organs: high incidence in Paris]. PMID- 2969545 TI - [Lactic acidosis caused by metformin. Value of intraerythrocyte levels]. PMID- 2969546 TI - [Does mastectomy still play a role in the treatment of cancer of the breast?]. PMID- 2969547 TI - [Effects of dietetic manipulations on the course of rheumatoid arthritis]. PMID- 2969548 TI - [Radiotherapy before amputation of the rectum for cancer. 100 cases]. AB - Between 1977 and 1982, 100 patients were treated with pelvic irradiation of 3000 rads given over a 3-week period for a carcinoma located in the lower third (51 cases) or the middle third (49 cases) of the rectum. The irradiation was well tolerated by all patients. The tumour disappeared in 4 patients who were not operated upon and survived for more than 5 years. Among the remaining 96 patients, 18 of whom had fixed lesions, rectal excision was performed in 87 (90.6 per cent) and was considered curative in 75 (78 per cent). Pathological examination of the surgical specimens revealed the absence of tumour in 4 cases. There were 2 post-operative deaths. After curative excision, the 5-year survival rate was 70 per cent and the locoregional recurrence rate was 8 per cent, with perineal recurrence in only one case. Patients treated with pre-operative irradiation have been compared with a historical series of patients who had abdominoperineal excision without irradiation. In the present series of irradiated patients, thromboembolic complications were more severe, and the problems encountered with healing of the perineum were solved by changing the surgical technique in order to close the perineal wound. The rectal tumours were smaller, with a smaller proportion of tumours of more than 5 cm in diameter (P less than 0.001), but there was no significant change in Duke's stage distribution. The 5-year survival rate was higher (70 per cent versus 55.3 per cent; P less than 0.05), and the 5-year locoregional recurrence rate was lower (8 per cent versus 18.4 per cent; P less than 0.05). PMID- 2969549 TI - [Changes in plasma binding protein of sex steroids and transcortin during the treatment of menopause with oral estrogens]. AB - Sex steroid binding protein (SBP) and transcortin (CBG) plasma concentrations were measured in 15 postmenopausal women before and during oral administration of estradiol 2 mg plus estriol 1 mg given alone for one month and in sequential combination with noresthisterone 1 mg for the following months. The results were compared with those obtained in a group of 13 premenopausal women who were studied during the early follicular phase or during administration of estroprogestagens. The oral administration of estrogens slightly increased CBG levels (56.1 +/- 11.4 vs 46.0 +/- 5.2 mg/l, P less than 0.05) which in 4 patient were higher than in premenopausal women. The mean SBP level was lower in postmenopausal women than in premenopausal women (1.02 +/- 0.40 vs 1.35 +/- 0.38 micrograms/dl, P less than 0.02), and SBP correlated negatively and significantly with the body mass index (r = 0.794, P less than 0.02). On average, SBP increased twofold during the estrogen treatment. In 6 patients the concentrations of estrogen-stimulated SBP were higher than the upper limit for premenopausal women. Lowered SBP levels were normalized during estrogen therapy. During estrogen substitution in the postmenopausal women, the mean E2 to SBP ratio (an index of free estradiol) was within the normal limits for premenopausal women. These results demonstrate that SBP is highly sensitive to oral estrogens. The increase in SBP is associated with a free E2 index which is within the physiological range of premenopausal women. The risk(s) or benefit(s) associated with the increase in SBP during estrogen therapy in postmenopausal women deserve to be evaluated by further investigations. PMID- 2969550 TI - [Conservative surgery of rheumatic mitral insufficiency in children. Remote results]. AB - Between 1977 and 1986, 38 children aged from 3.8 to 16.4 years underwent mitral valvuloplasty for symptomatic mitral valve incompetence of rheumatic origin. Two children (5.3 per cent) died in the immediate post-operative period, and 6 were lost sight of. Among the 30 children followed up for a mean period of 3.3 years, 6 were reoperated upon within 5 months of the valvuloplasty, 5 still present with a significant residual mitral valve incompetence and 9 have satisfactory clinical and echocardiographic results. Survival rate and survival rate without reoperation at 4 years were 93.8 per cent and 76.1 per cent respectively. Factors that correlated with the quality of long-term results were pre-operative left ventricular myocardial fibre shortening rate and surgical technique (poor results were obtained in children who had only one annuloplasty or none). PMID- 2969551 TI - [Prevention of deep venous thrombosis in arthroplastic surgery of the hip by the combination of heparinotherapy and the antithrombosis stocking]. AB - Two hundred and eighty-six patients were divided into two groups according to the presence or absence of major risk factors. The so-called "at risk" patients were treated with continuous post-operative heparin therapy combined or not with anti thrombosis stockings. The patients regarded as not at risk were divided into 3 subgroups allocated at random to different treatments based on subcutaneous heparin in effective or sub-anticoagulant doses with and without stocking. All patients underwent phlebography before and 7 days after the operation. The incidence of post-operative thrombosis was significantly lower in the 137 patients treated with heparin and stocking (10.22 per cent) than in the 131 patients treated with heparin alone (19.85 per cent). PMID- 2969552 TI - [Lipid evaluation in cerebral infarction in its early phase and in the 4th month. Practical consequences]. AB - Plasma lipid values studied in 30 subjects with stroke at days 1, 2 and 4 of the early phase indicated a significant decrease in total cholesterol, HDL cholesterol and apolipoprotein A1 from day 1 to 4. However, there was no significant difference between values at day 1 and during the 4th month of convalescence. Since some patients admitted to hospital are never followed up later in a hospital environment, it would seem advisable, for practical purposes, to perform plasma lipid analysis on the first day of admission. PMID- 2969553 TI - [Onset of convulsive crises with theophylline therapy at therapeutic doses]. AB - Six patients treated with theophylline, including five with advanced chronic obstructive pulmonary disease, developed generalized seizures with serum theophylline concentrations ranging from 7 to 21 mg/l. Cerebral computed tomographic scans showed a small sylvian infarction in one patient. In the other five patients nothing, except theophylline, could account for the seizures. There were no clinical signs of theophylline toxicity prior to the failures, all of which had a favourable outcome. Patients with acute exacerbations of chronic obstructive pulmonary disease seem to be prone to develop generalized seizures when treated with theophylline. PMID- 2969554 TI - [Hemoperfusion on charcoal and hemodialysis in acute poisoning caused by methotrexate]. AB - Acute methotrexate intoxication occurred in 4 patients despite adequate alkaline hyperhydration and classical folinic acid rescue. Three of these patients had no previous risk factor. Charcoal haemoperfusion with haemodialysis was promptly instituted and the methotrexate blood levels rapidly decreased, avoiding further renal damage and multisystemic involvement. Chemotherapy could subsequently be performed in 3 of the 4 patients without delay and toxicity. Charcoal haemofiltration appears to be an excellent treatment of methotrexate intoxication. PMID- 2969555 TI - [Lyme disease manifested by polyradiculoneuritis without hypercytosis of the cerebrospinal fluid]. PMID- 2969556 TI - [Myalgia induced by sotalol. A case]. PMID- 2969557 TI - [Primary hyperthyroidism associated with clear cell nephroepithelioma]. PMID- 2969558 TI - [Pure isolated Korsakoff syndrome after a gunshot wound]. PMID- 2969559 TI - [Cerebral thrombophlebitis associated with a protein C deficiency during pregnancy]. PMID- 2969560 TI - [A clinical test of digital sensitivity: the "usherette's sign"]. PMID- 2969561 TI - [Edematous syndromes caused by capillary hyperpermeability. Diffuse angioedema]. PMID- 2969562 TI - [Artificial nutrition in the alcoholic cirrhotic patient]. PMID- 2969563 TI - [Nipple discharge without palpable tumor. Experience of the Institut Curie from 1970 to 1984]. AB - Four hundred and ten women with serous or bleeding nipple discharge without palpable mass were treated by surgery in the Curie Institute between 1970 and 1984. The cancer rate in this group was 12.2 per cent. Galactography was of anatomical interest as it showed the canal at the origin of the discharge. Malignant cells were observed at cytological examination in only 17 per cent of intragalactophoric cancers. Surgical excision is necessary if the nipple discharge is uniporous and easily reproducible by areolar pressure, particularly in the post-menopausal period when the frequency of cancer is highest (17.7 per cent). PMID- 2969564 TI - [Superiority of fiberoptic inflation compared to fiberoptic bronchoscopy in the early treatment of atelectasis under mechanical ventilation]. AB - The purpose of this study was to compare the effectiveness of a simple technique of active lung reexpansion through a fiberoptic bronchoscope, using a large volume syringe, with that of the conventional fiberoptic bronchoscope suction. Thirty consecutive patients with atelectasis were divided at random into two groups: with and without active inflation. The two groups were similar as regards age, sex-ratio, side and area involved in atelectasis. The group without inflation was less hypoxaemic than the other group (PaO2 202 and 140 mmHg respectively). Early and lasting improvement in chest X-ray and arterial blood gases occurred in both groups, but patients were significantly more improved during fiberoptic inflation. No pneumothorax or haemoptysis was observed during the procedure. These data prompt us to recommend active inflation instead of fiberoptic bronchoscopy suction alone for the emergency treatment of patients with severe hypoxaemic atelectasis. PMID- 2969565 TI - [Identification of prognostic criteria of dilated cardiomyopathies associated with chronic alcoholism. 22 cases]. AB - Between November, 1978 and August, 1983, 22 patients aged from 28 to 65 years with idiopathic dilated cardiomyopathy diagnosed on the basis of clinical and haemodynamic criteria were investigated. All these patients, who consumed alcohol in excessive amounts, were followed up for 1 to 58 months. The overall mortality rate was 10 p. 100 at 4 months and 25 p. 100 at 58 months. Six patients were clinically improved with reduction of heart size (group A); 10 patients showed deterioration with 3 deaths (group C) and 6 patients followed an intermediate course (group B). When the characteristics of the groups were evaluated, it was found that compared to group C patients those in groups A and B had a lower cardiothoracic ratio: 0.56 +/- 0.04 (B) vs 0.64 +/- 0.06 (C) (P less than 0.02), a lower indexed end-diastolic diameter and systolic diameter on TM echo: 3.20 +/- 0.50 cm/m2 (A) vs 4.13 +/- 0.39 cm/m2 (C) (P less than 0.02), and 2.72 +/- 0.37 cm/m2 (A) vs 3.57 +/- 0.47 cm/m2 (C) (P less than 0.02) respectively, and a lower indexed end-diastolic volume as evaluated by angiography: 121 +/- 61 ml/m2 (A) vs 202 +/- 65 ml/m2 (C) (P less than 0.06). Dilated cardiomyopathy associated with excessive alcohol consumption has a better prognosis when the patients stop drinking and when their heart dilatation is mild to moderate. PMID- 2969566 TI - [Nosocomial septicemia and pseudobacteremia caused by Serratia marcescens]. AB - An epidemiological survey was carried out which included a dual epidemic of septicaemia and pseudo-bacteremia caused by Serratia marcescens. The survey enabled 15 septicaemias and 43 pseudobacteremias to be detected in a regional hospital between March and August, 1983. Two mishandlings were at the origin of the outbreak: citrated tube normally reserved for coagulation tests were severely contaminated by Serratia marcescens, and inaccurate samplings had been made. Once the mechanisms of contamination were found, specific preventive measures put an end to the epidemic. The authors insist on the need for uncontaminated tubes and citrate solutions and for the development of precise sampling methods which are essential to avoid the occurrence of pseudo-bacteremia or septicaemia. It is important to detect such epidemics at an early stage by an efficient control of nosocomial infections, thus avoiding their extension. PMID- 2969567 TI - [Systemic effects of beta blockaders used in collyria]. AB - Eye-drops containing beta-blockers are widely used by ophthalmologists in the treatment of chronic glaucoma, since they reduce the intraocular pressure by about 25-30 per cent. This treatment is usually well tolerated but not devoid of systemic effects. The eye-drops are rapidly removed by the lacrymal system, and 80 per cent of the drug is absorbed by the nasal mucosa. It is then taken up by the blood stream in low but not negligible concentrations, and since the liver is by-passed the active substance acts directly on the target organs. Side-effects are similar to those observed after oral administration, despite much lower plasma levels; although rare in routine clinical practice, they may result in decompensation of a fragile cardiac or respiratory function. This does not mean that beta-blocker eye-drops should not be prescribed to patients with chronic glaucoma, but attention must be paid to the patient's cardio-respiratory status and to a possible concomitant treatment with drugs that depress myocardial conductivity or contractility. PMID- 2969568 TI - [Treatment of prolapse of the rectum in the young adult with the so-called Delorme procedure]. PMID- 2969569 TI - [Value of procuring the right kidney with the infrarenal inferior vena cava for transplantation]. PMID- 2969570 TI - [Third-degree atrioventricular block in typhoid fever]. PMID- 2969571 TI - [Gram-negative septicemia difficult to culture and identify after a dog bite]. PMID- 2969572 TI - [Joint diseases in hemodialysed patients. The extrarenal epuration method is not involved]. PMID- 2969573 TI - [Digestive bypass syndrome observed in hemorrhagic rectocolitis]. PMID- 2969575 TI - [Changes in sharing injection equipment among drug addicts]. PMID- 2969576 TI - [Analysis and separation of cells by flow cytometry]. PMID- 2969574 TI - [Calcium channel blockers in the surgery of pheochromocytoma]. PMID- 2969577 TI - [Do digestive complications of corticotherapy exist?]. PMID- 2969579 TI - [Perforated gastroduodenal ulcers. Treatment by peritoneal dialysis. 72 cases]. AB - Seventy-two perforated gastroduodenal ulcers were treated by an original method aimed at avoiding emergency surgery, which consists of peritoneal dialysis associated, during 3 days, with gastric aspiration. Over a 3-year period this method was applied to all patients admitted with a perforated ulcer. The ulcer was revealed by the perforation in 25 p. 100 of the cases, and 90 p. 100 of the patients had pneumoperitoneum. Contrast radiography with Gastrografine (sodium and meglumine amidotrizoate) located the perforation. Fifteen patients had another disease which made the prognosis worse. The time elapsed between perforation and treatment was 15 hours on average. Peritoneal fluid infection was present in 30 p. 100 of the cases. The outcome was favourable in 69 patients (96 p. 100). One patient died of pulmonary embolism, another was operated upon on the 4 th post-perforation day for a bleeding ulcer and a third patient with giant gastric ulcer developed subphrenic abscess. This method seems to be indicated in patients at high surgical risk (elderly people or people with severe underlying disease), and in young patients with perforated acute ulcer. In chronic ulcers, peritoneal dialysis ties the patient over the first hours, thus enabling radical surgery to be electively performed. In perforations seen after 24 hours, it helps in supporting the patient prior to surgery. The method in contra-indicated in gastric ulcers. PMID- 2969578 TI - [Sero-epidemiology of viral hepatitis A, B and delta in Martinique]. AB - In view of the ethnic and geographical peculiarities of the French department of Martinique and of the endemic character of hepatitis in tropical countries, we studied the prevalence of infections with hepatitis A, B and delta viruses in that region. A group of 10,109 blood donors and a group of about 100 patients were selected on account of their liver symptoms. As regards hepatitis A, the study of the 2 groups was completed by a sero-epidemiological survey of 509 children and teenagers aged from 1 to 18 years. The prevalence of the HB antigen among blood donors was 1.3 per cent, i.e. about 10 times higher than in Europe and 7 times lower than in hyperendemic tropical areas. It was 2.5 times higher in the male than in the female population; 84 per cent of HBs-positive donors had anti-HBe antibodies, 9 per cent had HBe antigen and 7 per cent had neither one nor the other. This distribution is coherent with a population of symptomless carriers. The prevalence of anti-HBs-positive sera was 34 per cent as against 70 95 per cent in highly endemic countries and 4-20 per cent in Europe; 1.9 per cent of the HBs donors studied and 8.8 per cent of the patients had anti-delta antibodies; 11 of the 13 anti-delta-positive subjects had anti-HBe antibodies and 2 had neither HBe nor anti-HBe. Between the ages of 1 and 3 years very few anti HAV-positive subjects were observed. From 3 to 10 years, the percentage of seroconversions increased moderately. Between 10 and 20 years, the number of positive cases increased considerably reaching 67 per cent at 20 years and 100 per cent at and above 45 years. Among the patients, 97 were positive for IgG (96 per cent) and only 5 for IgM (4 per cent). PMID- 2969580 TI - [Hepatic complications after high-dose chemotherapy and bone marrow autograft in solid tumors in children]. AB - Two-hundred and thirty-six courses of high-dose chemotherapy with bone marrow transplantation for solid tumors in children were reviewed in order to assess the incidence of hepatic abnormalities and their etiology. Bacterial and fungal hepatitis were excluded from this study. Hepatic complications occurred after 43 courses (20 per cent) in 39 patients. Four patients had hepatic abnormalities after two consecutive courses. Predisposing factors were found to be conditioning regimen including busulfan and previous high dose chemotherapy. Veno-occlusive disease occurred in 11 children and had a fatal outcome in 4. In 9 children, hepatitis was a symptom of diffuse viral infection. In the other 23 courses, hepatitis was isolated but it was difficult to determine whether its etiology was viral (cytomegalovirus, Epstein-Barr virus, adenovirus, HBs, non A non B) or toxic. In these latter forms, the outcome was always good. PMID- 2969581 TI - [Allergy to Hymenoptera. Incidence and degree of public knowledge]. AB - The prevalence of systemic reactions to hymenoptera stings in adults is unknown. To elucidate this point we interviewed 2.067 persons aged from 20 to 60 years who attended a medical screening centre. We asked them whether they had ever experienced a reaction this kind, what they knew about it and what they did, and we performed skin tests to hymenoptera venoms in those who gave a positive answer. The proportion of subjects who had had at least one type of systemic reaction after being stung was 1.2 per cent; 0.9 per cent of the subjects had both a history of reaction and a positive skin test. This figure is in keeping with those obtained in surveys carried out among children, but is does not tally with the mortality figures which probably are grossly under-estimated. The general population is not adequately informed of possible systemic reactions to hymenoptera stings, so that people often fail to consult, and the practitioners themselves do not advise their patients to see an allergologist for evaluation. Thus, adequate information of the general population and the physicians is badly needed. PMID- 2969582 TI - [Endoscopic transvesicoparietal exteriorization of ureteral catheters]. AB - Endoscopic introduction of catheter into the ureter for drainage has long been a common procedure, and recent advances in endoscopic urology have widened its indications. The ureteral catheter is usually exteriorized through the urethra and attached to the urethral catheter, but with this technique it tends to descend spontaneously and to create infections which are a possible cause of ureteral stenosis. A technique of endoscopic transvesicoparietal exteriorization of ureteral catheters without incision has been developed in 1982. The problems encountered in exteriorizing the ureteral catheter through the lumen of a suprapubic trocar have now been solved by using a bipodal forceps introduced into the trocar. PMID- 2969583 TI - [Critical study of spinal fluid lactic acid levels in the etiological diagnosis of meningitis in children. Verification of high spinal fluid lactic acid levels in lymphocytic meningitis caused by mumps]. PMID- 2969584 TI - [Non-Hodgkin's malignant lymphoma associated with monoclonal immunoglobulin in two brothers]. PMID- 2969585 TI - [Thrombocytopenia in leptospirosis. Role of anti-platelet antibodies]. PMID- 2969586 TI - [Glomerular involvement secondary to portacaval anastomosis for congenital hepatic fibrosis]. PMID- 2969587 TI - [Diagnosis of erosive discopathies in lumbar arthrosis. Value of magnetic resonance imaging]. PMID- 2969588 TI - [Thrombolysis in the home. Analysis of contraindications]. PMID- 2969590 TI - [Acute hepato-nephritis in an alcoholic following a therapeutic dose of paracetamol]. PMID- 2969589 TI - [Listerial rhombencephalitis in a non-immunodepressed patient. Fatal development despite early antibiotherapy]. PMID- 2969591 TI - Translation initiation controls the relative rates of expression of the bacteriophage lambda late genes. AB - The late operon of bacteriophage lambda contains the genes encoding the morphogenetic proteins of the phage. These genes are transcribed equally from the single late promoter. Although the functional half-lives of the mRNA for the various genes of this operon vary less than 2-fold, their relative rates of expression have been shown to vary by nearly 1000-fold. This variation could result from differing rates of translation initiation, from overlapping upstream translation, or from differential elongation rates due to the presence of codons for which the corresponding tRNAs are rare. To distinguish between these possibilities, we have cloned sequences surrounding the initiator codons of several of these genes and measured their ability to drive synthesis of hybrid lambda-beta-galactosidase proteins. The rates of expression of the hybrid genes thus produced correlate very well with the natural rates of expression of the corresponding phage genes, suggesting that the rate of initiation of translation controls the relative expression rates of these genes. PMID- 2969592 TI - Association of U2, U4, U5, and U6 small nuclear ribonucleoproteins in a spliceosome-type complex in absence of precursor RNA. AB - Small nuclear ribonucleoprotein particles (snRNPs) associate to form multi-snRNP complexes during splicing of mRNA precursors. A vast majority of the three snRNPs U4, U5, and U6 are present in a nuclear extract in a single complex, while U1 and U2 snRNPs exist as separate particles. Under conditions optimal for splicing in vitro the U4-U5-U6 (U4/5/6) complex dissociates to release free snRNPs, suggesting that the interactions between its components are dynamic. Several forms of splicing complexes assemble on precursor RNA during splicing in vitro. One of these forms, spliceosome B, contains U2, U4, U5, and U6 snRNPs bound to the precursor RNA. This same set of snRNPs associates efficiently in the absence of precursor RNA during incubation of the extract at high salt concentration. Formation of this U2-U4-U5-U6 (U2/4/5/6) complex, the pseudospliceosome, suggests that the basic structure of the spliceosome is specified by snRNP-snRNP interactions. PMID- 2969593 TI - Activation and expansion of hapten- and protein-specific T helper cells from nonsensitized mice. AB - Hapten- and protein-antigen-specific T helper cells are usually expanded in vitro from lymphocytes obtained from sensitized animals. In this paper we report on the primary activation and proliferation in vitro of T helper cells from nonsensitized animals by using syngeneic cultured epidermal Langerhans cells as a source of potent antigen-presenting cells. The primary in vitro proliferation was blocked with monoclonal antibodies to Ia molecules, to lymphocyte function associated antigen 1 (LFA-1), and to L3T4. T helper cell populations sensitized in vitro to haptens and protein antigens showed hapten- and antigen-specific proliferation when restimulated in vitro with spleen cells. Besides its experimental usefulness, in vitro generation of syngeneic specific T helper cells may afford possibilities for adoptive immunotherapy. PMID- 2969595 TI - [The partially handicapped person in society]. PMID- 2969594 TI - cDNA heterogeneity suggests structural variants related to the high-affinity IgE receptor. AB - The high-affinity IgE receptor present on mast cells and basophils is responsible for the IgE-mediated activation of these cells. The current model for this receptor depicts a four-subunit structure, alpha beta gamma 2. A cDNA for the alpha subunit was recently cloned and predicts a structure consisting of two homologous extracellular domains, a transmembrane segment, and a cytoplasmic tail. Using a synthetic oligonucleotide corresponding to the amino-terminal sequence of the alpha subunit, we identified a number of cDNA clones from a rat basophilic leukemia cell cDNA library. Nucleotide sequencing established four different forms of cDNA: one is nearly identical to the published cDNA; the second differs from the first in the 5' untranslated sequence; the other two forms use either one or the other of the 5'-end sequences as above and lack 163 base pairs in the region coding for the second extracellular domain. RNase protection analysis with radioactive RNA probes established the heterogeneity of rat basophilic leukemia cell mRNA with regard to both the 5' and the internal sequences. Our results suggest the existence of at least four different protein forms related to the alpha subunit of the high-affinity IgE receptor. PMID- 2969596 TI - Innervation of the rectus abdominis muscle: implications for rectus flaps. AB - The usefulness of leaving lateral strips of the rectus abdominis muscle in place during a transverse rectus abdominis musculocutaneous (TRAM) flap procedure is questioned. Since textbooks do not agree on the course of the intercostal nerves in the rectus fascia and no precise description is given of the exact site of penetration of the nerves in the rectus muscle, six fresh cadavers were dissected. It has been observed that the nerves enter the deep face of the muscle in its middle portion. Lateral parts of the muscle are consequently denervated during a transverse rectus abdominis musculocutaneous flap, which preserves them. This has been confirmed by CT scan of the abdominal wall in 10 patients 2 to 37 months after a transverse rectus abdominis musculocutaneous flap. In these patients, a progressive fibrosis and disappearance of the remaining muscle could be demonstrated. It is concluded that a partial taking of the rectus abdominis muscle does not preserve its muscular function. PMID- 2969597 TI - [Proteinase participation in chromosome radiation damage. Research on the action of phenylmethylsulfonyl fluoride]. AB - Phenylmethylsulfonyl fluoride, a chromatin proteinase inhibitor, caused a nearly twofold diminution of the cytogenetic injury and a twofold increase in the rate of DNA repair in gamma-irradiated (3-15 Gy) Chinese hamster fibroblasts. The effect of the inhibitor was mainly exhibited by a rapidly repaired (for 15-20 min) component of the cytogenetic damage. A simultaneous treatment with phenylmethylsulfonyl fluoride and nicotinamide did not influence the effect of the proteinase inhibitor under study. The results obtained are indicative of poly (ADP-ribosylation)-independent contribution of chromatin proteinases to radiation induced chromosome mutagenesis. PMID- 2969598 TI - [A mathematical model of postradiation autoimmunity]. AB - A mathematical model of cellular autoimmune process in exposed mammals was developed. In terms of this model a study was made of the dependence of the autoimmunity kinetics on radiation dose and radiosensitivity of autologous tissues. The model simulates the experimentally observed dynamics of autoimmune diseases. PMID- 2969599 TI - [A love affair with phi X174. 1]. PMID- 2969600 TI - Immune complex induced pancreatitis: effect of BN 52021, a selective antagonist of platelet-activating factor. AB - A model of acute pancreatitis was developed by induction of an immune complex mediated hypersensitivity reaction in rats. This acute inflammatory reaction was characterized by intense interstitial edema, neutrophil infiltration and margination, and congestion of small vessels whereas serum amylase levels remained unchanged. Microscopic examination of the pancreatic tissue revealed the presence of immune complex deposition around blood vessels and ducts. Vascular permeability, as measured by Evan's blue extravasation increased by 6 fold. In addition, circulating platelets dropped to 50% of normal levels. Injection of platelet-activating factor (PAF) in the peritoneal cavity of rats also produced an increase in vascular permeability in the pancreas. A selective PAF-antagonist, BN 52021 reduced by approximately 50% the increase in vascular permeability produced by immune complex in the pancreas as well as that elicited by intraperitoneal injection of PAF. These results suggest that PAF plays a role in the pathological manifestations of immune complex-mediated pancreatitis. PMID- 2969601 TI - [Percutaneous transluminal angioplasty after renal transplantation]. AB - Renal artery stenosis is a frequent complication of kidney transplantation (10%). Percutaneous transluminal angioplasty (PTA) has recently been proposed as a potential therapeutic procedure. Twelve transplant patients with arterial stenosis underwent PTA. The procedure was successful in 10 cases (83.3%). Restenosis occurred in 2 patients (16.7%); both of them underwent PTA successfully. No complications occurred. A considerable improvement in glomerular filtration rate and a reduction in high blood pressure were observed in all patients after successful PTA. The authors believe PTA to be the therapy of choice in the treatment of arterial stenoses in kidney transplant patients. PMID- 2969602 TI - Autoantibodies. AB - The diagnosis of SLE is associated with an enlarging assortment of autoantibodies. The presence or absence of particular antibodies influences the confidence with which this diagnosis is made. It is the presence of autoantibodies and the deposit of immunoglobulin that has led to the general conclusion that lupus is an autoimmune disease. PMID- 2969603 TI - Facet joint disorders and their role in the production of back pain and sciatica. AB - This article, say the authors, serves as a reminder that the posterior articulations of the lumbar spine are key elements in the production of low back pain and sciatica. PMID- 2969604 TI - Local antibiotic treatment of soft-tissue infections with gentamicin-PMMA chains. PMID- 2969605 TI - [A case of repeat percutaneous transluminal coronary angioplasty for recurrent aorto-coronary bypass graft stenosis]. PMID- 2969606 TI - [Atrial natriuretic factor in man: physiopathological implications]. PMID- 2969607 TI - [Evaluation criteria of chronic fetal distress. Respective role of a Doppler blood velocimetry study of the umbilical and uterine arteries]. PMID- 2969608 TI - [Fetal biophysical profile used with or contrasted to the placental resistance index]. PMID- 2969609 TI - [Ovarian cysts: ultrasound-guided puncture, celioscopy or laparotomy?]. AB - The authors report a series of 262 "ovarian tumors" operated on between 1982 and 1986. The place of "primary" laparoscopy went from 79% in 82-84 to 83% in 85 and 85% in 86 and the place of "pure" laparoscopic treatment went from 39% to 57% and 60%, respectively, excluding patients with an obvious cancer, patients over 40 and pregnant women over 16 weeks, in whom a primary laparotomy is indicated. The indications of the laparoscopic treatment may be carried further. And there is a place for a tap under ultrasonographic control. Various approaches are presented. PMID- 2969610 TI - Gastric blood flow in patients with gastric ulcer measured by endoscopic laser Doppler flowmetry. AB - Gastric blood flow has been studied endoscopically by laser Doppler flowmetry in 15 patients with chronic gastric ulcer. In all patients the ulcer was located at the lesser curvature of the corpus. The blood flow measured in this area was decreased compared with healthy controls, whereas the flow values in other parts of the stomach were similar to those of controls. In the ulcer bed very low blood flow values were measured. The blood flow of the ulcer margin was similar to that of other parts of the lesser curvature. After 4-6 weeks' treatment with the H2 receptor antagonist cimetidine, the ulcer was healed in about 70% of the patients. The blood flow measured at the lesser curvature and in the ulcer area was still low and of the same magnitude as the corresponding values of the first measurement. After 4 months the ulcer was healed in all but one patient. The blood flow of the lesser curvature had increased significantly, whereas decreased blood flow was measured at the distal part of the greater curvature. In the area where the ulcer was located, very low flow values were still measured. This finding may explain why ulcer recurrence usually occurs at the site of the primary ulcer. PMID- 2969611 TI - Leukaemic B cells from patients with chronic lymphocytic leukaemia suppress immunoglobulin production by lymphocytes from normal donors. AB - We observed that highly purified E-rosette-negative largely leukaemic B cells from 9 out of 15 patients with chronic lymphocytic leukaemia (CLL) significantly suppressed immunoglobulin production by mixtures of T4 and B cells from normal donors in the presence of pokeweed mitogen (PWM). This suppression by leukaemic B cells was concentration-dependent. Addition of equal numbers of B cells from normal donors to the mixtures of normal T4 and B cells increased, or had no effect on the production of IgM, IgA, and IgG. Treatment of purified largely leukaemic B cells from patients with CLL with either the anti-B1 or anti-Leu 1 monoclonal antibody plus complement abolished their ability to suppress immunoglobulin production. In contrast, treatment with either the anti-Leu 5 or the OKM1 monoclonal antibody plus complement had no effect on the suppression. These results suggest that leukaemic B cells from certain patients with CLL may exhibit, or can be induced to exhibit, immunosuppressive properties. PMID- 2969612 TI - Inefficient binding of IgM immune complexes to erythrocyte C3b-C4b receptors (CR1) and weak incorporation of C3b-iC3b into the complexes. AB - The binding of soluble complement-reacted IgM immune complexes (IC) to erythrocyte (E) C3b-C4b receptors (CR1) and the incorporation of C3b-iC3b into solid phase IgM-IC was investigated. The optimal binding of liquid phase IgM-IC to E-CR1 was obtained with IC formed at moderate antibody excess, but the binding was low (2-3%) when compared to the binding of the corresponding IgG-IC (50-60%). Solid phase IC were prepared by coating microwells with heat-aggregated bovine serum albumin (BSA) followed by incubation with rabbit IgM anti-BSA antibody. The IC were reacted with human serum at 37 degrees C. The binding of C3b-iC3b was determined by use of biotinylated F(ab')2 antibodies to C3b-C3c and avidin coupled alkaline phosphatase. The incorporation of C3b-iC3b into solid-phase IgM IC increased when increasing amounts of IgM antibody were reacted with the antigen. The binding reaction was slow, reaching a maximum after about 2 h at 37 degrees C. The binding of C3b-iC3b to the IgM-IC was remarkably inefficient when compared to the incorporation into IgG-IC reacted with the same amounts of BSA precipitating antibody. PMID- 2969613 TI - Dynamics of cytotoxic T lymphocyte precursors in vivo assessed by change in the radiation sensitivity. Evidence for development of radiation-sensitive memory cells without clonal expansion. AB - The dynamics of cytotoxic T lymphocyte precursors (CTL-p) in mice injected with allogeneic spleen cells (SC) was studied with special reference to changes in their radiation sensitivity. Whole-body 400 rad X-ray irradiation of allo-SC primed and unprimed mice virtually abolished the capacity of their SC to proliferate and to generate CTL in primary or secondary mixed leucocyte culture (MLC). However, the impaired ability of SC to generate CTL in the primary MLC was restored by interleukin 2 (IL-2). This showed that helper cells whose activity was replaceable with IL-2 (IL-2-producing cells) were functionally more radiation sensitive than CTL-p in unprimed mice. In contrast, the radiation-impaired activity in secondary MLC was not restored by IL-2, suggesting that memory CTL-p in allo-SC-primed mice were unexpectedly sensitive to radiation. The D37 values determined from the percentage of residual CTL-p activity of SC in bulk cultures 1 day after irradiation were 525 rad for virgin CTL-p and 75 rad for memory CTL p. Further studies demonstrated that the radiation-sensitive memory CTL-p were generated from relatively radiation-resistant precursors, largely independent of radiation-sensitive IL-2-producing cells and of cellular proliferation. The mean frequency of CTL-p in SC measured by limiting dilution assay was not significantly increased by the priming. This supports our conclusion that the development of the memory CTL-p activity in allo-SC-primed mice did not depend on clonal expansion. Whole-body 400 rad-irradiation reduced the frequency of CTL-p in SC from unprimed mice to 1/2-1/3 and that in SC from allo-SC-primed mice to 1/8-1/15. This supports the view that the majority of radiation-resistant virgin CTL-p functionally mature to radiation-sensitive memory CTL-p without cellular proliferation in allo-SC-primed mice. PMID- 2969614 TI - Kinetics of endotoxin release by gram-negative bacteria in the intestinal tract of mice during oral administration of bacitracin and during in vitro growth. AB - The release of endotoxin by gram-negative bacteria was studied during bacitracin induced intestinal colonization in C3H/Law mice and during in vitro growth. The "free" endotoxin concentration was determined by the Limulus amebocyte lysate test in faecal and in culture supernatants, respectively. After oral administration of bacitracin for 2 days a significant (p less than 0.001) increase of the faecal concentration of gram-negative bacteria of 3-4 logs accompanied a significant (p less than 0.001) increase of the faecal endotoxin concentration from 10(1.8 +/- 0.2) to 10(3.3 +/- 0.2) micrograms endotoxin/g faeces. In vitro, however, an increase of the concentration of gram-negative bacteria of 3-4 logs resulted in a 3-4 log increase of the concentration of endotoxin during the exponential and early stationary growth phase. The faecal endotoxin level after 8 days of bacitracin treatment dropped to a value not significantly different from the initial value, regardless of a high faecal level of gram-negative bacteria. Endotoxin determination by the Limulus amebocyte lysate test appeared to be unaffected by the amount of bacitracin present in faecal preparations after oral administration of this antibiotic. In addition, the in vitro release of endotoxin was not influenced by bacitracin. From these results we conclude, that "free" endotoxin is a product of extensive proliferation of gram-negative bacteria. Therefore, the intestinal endotoxin level does not necessarily correlate with the level of gram-negative bacteria, but corresponds with the proliferative activity of these bacteria. PMID- 2969615 TI - Head injury. Integrated rehabilitation approach, optimal adaptation to disability. AB - Comprehensive rehabilitation of head injury means that all somatic, psychological and social consequences of the accident must be taken into account in order to plan the most appropriate individual therapeutic program. A review is given of the epidemiology, the different types of head injuries and the management in the acute stage and the rehabilitation stage with special reference to severe and moderate head injuries as well as minor head injuries and post-concussion syndrome. PMID- 2969616 TI - [Cardiovascular radiology]. AB - Some fifty years ago, the first attempts to visualize cardiac cavities by X-ray were reported, and a few years later right heart catheterization was performed by a physician on himself. After the Second World War, major new developments in cardiothoracic surgery, anesthesiology and extracorporeal circulation allowed surgeons to investigate a new field: cardiac surgery. For successful surgery it was mandatory to establish an exact preoperative diagnosis of congenital and acquired cardiopathies; physicians and engineers developed catheterization techniques which allowed hemodynamic data to be collected. Sophisticated radiologic equipment was designed and cardiovascular radiology was born. A quarter-century after the first cardiac surgery, a new era of treatment for cardiovascular diseases has been opened up with the development of interventional radiology. Gastrointestinal and pulmonary hemorrhages can be treated by occlusion of the feeding artery. Angina pectoris, impending infarct, hypertension and claudication can nowadays be treated not only by vascular surgery but also by interventional radiology using techniques such as balloon angioplasty. Percutaneous revascularization techniques are now under study for the benefit of the patient whose cardiovascular disease can be cured or improved with a hospital stay of one or two days, without surgical intervention or anesthesia. PMID- 2969617 TI - [Immunodiagnosis of leishmaniasis in dogs by ELISA and mini-Western blot]. PMID- 2969618 TI - cDNA expression cloning of the IL-1 receptor, a member of the immunoglobulin superfamily. AB - Interleukin-1 alpha and -1 beta (IL-1 alpha and IL-1 beta) are cytokines that participate in the regulation of immune responses, inflammatory reactions, and hematopoiesis. A direct expression strategy was used to clone the receptor for IL 1 from mouse T cells. The product of the cloned complementary DNA binds both IL-1 alpha and IL-1 beta in a manner indistinguishable from that of the native T cell IL-1 receptor. The extracellular, IL-1 binding portion of the receptor is 319 amino acids in length and is composed of three immunoglobulin-like domains. The cytoplasmic portion of the receptor is 217 amino acids long. PMID- 2969620 TI - [Dynamics of morbidity with temporary disability in various branches of industry during the 10th and 11th five-year periods]. PMID- 2969621 TI - [Teaching a course on the history of the CPSU with reference to professional orientation in a medical institute]. PMID- 2969619 TI - Synthetic CD4 peptide derivatives that inhibit HIV infection and cytopathicity. AB - Synthetic peptide segments of the CD4 molecule were tested for their ability to inhibit infection of CD4+ cells by the human immunodeficiency virus (HIV) and to inhibit HIV-induced cell fusion. A peptide mixture composed of CD4(76-94), and synthesis side products, blocked HIV-induced cell fusion at a nominal concentration of 125 micromolar. Upon high-performance liquid chromatography, the antisyncytial activity of the peptide mixture was found not in the fraction containing the peptide CD4(76-94) itself, but in a side fraction containing derivatized peptide products generated in the automated synthesis. Derivatized deletion and substitution peptides in the region CD4(76-94) were used to demonstrate sequence specificity, a requirement for benzyl derivatization, and a core seven-residue fragment required for antisyncytial activity. A partially purified S-benzyl-CD4(83-94) peptide mixture inhibited HIV-induced cell fusion at a nominal concentration of less than or equal to 32 micromolar. Derivatized CD4 peptides blocked cell fusion induced by several HIV isolates and by the simian immunodeficiency virus, SIV, and blocked infection in vitro by four HIV-1 isolates with widely variant envelope gene sequences. Purified CD4(83-94) dibenzylated at cysteine 86 and glutamate 87 possessed antisyncytial activity at 125 micromolar. Derivatization may specifically alter the conformation of CD4 holoreceptor peptide fragments, increasing their antiviral efficacy. PMID- 2969622 TI - [Physician-Bolshevik V.P. Lebedeva (1881-1968) and her contribution to the development of the Soviet system of maternal and child care]. PMID- 2969623 TI - [History of scientific medical societies in Ekaterinburg (Sverdlovsk)]. PMID- 2969624 TI - [Gelatinous transformation of the bone marrow. Cyto-histologic, histochemical and ultrastructural study of a case]. PMID- 2969625 TI - Disability research. PMID- 2969626 TI - Identifying the disabled: concepts and methods in the measurement of disability. By Lawrence D. Haber, 1967. PMID- 2969627 TI - The iliohypogastric and ilioinguinal nerves. Distribution in the abdominal wall, danger areas in surgical incisions in the inguinal and pubic regions and reflected visceral pain in their dermatomes. AB - The courses of the iliohypogastric and ilioinguinal nerves were studied in 44 adult human cadavers, in order to clarify their relations to incisions in the abdominal wall in appendectomy, inguinal hernial repair, caesarean section and lumbar nephrectomy. If either of these nerves is trapped during suturing of the abdominal layers, especially after inguinal hernia repair and appendectomy typical nerve irritation in the inguinal region is observed. To avoid cutting the anterior branches of the iliohypogastric and ilioinguinal nerves in appendectomy, incisions should be placed at a distance of not less than 3 cm from the anterior superior iliac spine. In inguinal hernial repair, after the external oblique aponeurosis has been opened, the ilioinguinal nerve should be displaced from the spermatic cord cranially. In performing a lower paramedian incision (Lennander) and Pfannenstiel's suprapubic incision, the iliohypogastric nerve will be spared if the incision passes at least 5 cm cranial to the inguinal ligament. During oblique lumbar incision for nephrectomy (Bergmann-Israel) the iliohypogastric nerve can easily be found in the middle third of the lateral margin of the quadratus lumborum muscle. The nerve should be displaced carefully downwards. Positional changes of the kidney or ureter, perinephric inflammation, etc. are often referred to the skin areas (Head, Mackenzie) of the iliohypogastric and ilioinguinal nerves. PMID- 2969628 TI - Atlantoaxial dislocation presenting as sudden onset of quadriplegia in Down's syndrome. AB - A sudden onset of quadriplegia and dyspnea occurred in a 4-year-old girl with Down's syndrome, who had been noted to have atlantoaxial dislocation. She underwent cervical immobilization initially with a halo-vest and subsequently by posterior fusion, resulting in significant functional recovery. The catastrophic myelopathy that occurred in this patient emphasizes the necessity of treatment even for asymptomatic atlantoaxial instability in some individuals with Down's syndrome. PMID- 2969629 TI - Hormonotherapy of meningiomas with medroxyprogesterone acetate. Immunohistochemical demonstration of the effect of medroxyprogesterone acetate on growth fractions of meningioma cells using the monoclonal antibody Ki-67. AB - The effect of medroxyprogesterone acetate (MPA) on growth fractions of ex vivo meningiomas is demonstrated in using the Ki-67 monoclonal antibody in three cases of meningiomas operated on in two stages and in meningioma specimens from a group of eight patients operated on in one single stage after MPA therapy. Growth fractions in samples from five meningioma patients not treated with MPA were determined for comparison. In the three cases of two-stage operation of the tumors, the percentage of Ki-67-positive cells in meningioma tissue was lower by a factor of 6, 5, and 3, respectively, after MPA therapy. In meningioma specimens from patients receiving no MPA therapy, Ki-67-positive cells were present in 1.02 +/- 0.48%; in samples from MPA-treated tumors the percentage of Ki-67-positive cells was 0.41 +/- 0.40 (different at p less than 0.02 [Wilcoxon's test]). In comparison to our previously published data on untreated meningiomas analyzed for progesterone receptors (PR), MPA significantly reduced the PR activity. There was no obvious correlation between PR activity and potential suppression of the tumor growth fraction. It is concluded that MPA is attractive because it reduces the growth fractions of most meningiomas and might be suitable for adjuvant hormonotherapy. PMID- 2969630 TI - Permanent rat cardiac allograft survival induced by ultraviolet B-irradiated donor lymphocytes and peritransplant cyclosporine. AB - This study examines the effect of pretreatment with 10(8) ultraviolet B irradiated donor leukocytes (UV-DL) with or without peritransplant cyclosporine (CyA) treatment (20 mg/kg on days 0, +1, and +2 relative to transplantation) on rat cardiac allograft survival across major histocompatibility loci. A single UV DL pretreatment on day -3 or -7 (before transplantation) significantly prolonged survival of heart allografts from Wistar-Furth rats (W/F) in Lewis recipients from 6.8 +/- 0.8 days to 18.4 +/- 2.1 and 17.6 +/- 1.5 days (p less than 0.001), respectively. Multiple UV-DL infusions on days -14 and -7 increased the mean survival time to 20.0 +/- 0.9 days (p less than 0.001). Similarly, UV-DL infusion on day -3 or -7 significantly prolonged the mean survival time of heart allografts from ACI rats in Lewis rats. A single or multiple UV-DL infusions combined with peritransplant CyA led specifically to permanent W/F cardiac allograft survival (more than 200 days) in all recipients. Similarly, UV-DL infusion combined with peritransplant CyA led to indefinite survival of ACI cardiac allografts in two thirds of Lewis recipients. Adoptive transfer of splenocytes from long-term recipients of cardiac allografts, which specifically prolonged donor test grafts in syngeneic hosts, suggests that unresponsiveness to cardiac allografts is, in part, dependent on suppressor cells. This study emphasizes the importance of UV irradiation of DLs in the modulation of alloreactivity and the induction of donor-specific unresponsiveness in adult animals. PMID- 2969631 TI - Systemic effects of prosthetic vascular graft implantation. AB - The effect of prosthetic vascular graft implantation on systemic complement activation, as well as leukocyte and platelet counts, was studied in rabbits. Animals underwent aortic clamping and declamping alone (group I) or with implantation of an externally supported PTFE graft (group II) or preclotted knitted Dacron graft (group III). Fifteen minutes after declamping, mild activation of complement reflected as a fall in CH50 to 72% of preoperative baseline values was observed in group I animals. Comparatively greater activation was seen in group II (43% baseline) and group III (28% baseline) animals. The differences between groups I and II (p less than 0.05) and I and III (p less than 0.01) were significant. The drop in platelet count was greater in group II (66% baseline) and group III (53% baseline) animals compared with group I (86% baseline). Neutrophil counts decreased in group I animals (75% baseline), but, again, this fall was more marked in group II (35% baseline) and group III (43% baseline) animals. The difference between groups I and II was significant (p less than 0.05). Similarly, monocyte count decreased in group I animals (80% baseline), which was also more marked in group II (32% baseline) and group III (31% baseline) animals. The differences between group I and groups II and III were significant (p less than 0.05 for each). These trends persisted throughout the first postoperative hour. These results suggest that the blood-surface interactions produced systemic effects in vivo. PMID- 2969632 TI - Interferon inhibition of IL-2-mediated lymphocyte proliferation. AB - Interleukin-2 (IL-2) combined with hybrid human alpha interferon (IFN-alpha A/D) mediates enhanced tumor regression compared with either agent alone. To elucidate the underlying mechanism of interaction of IL-2, with its known ability to induce expansion of T lymphocytes, and IFN-alpha A/D, with its antiproliferative activity, we studied splenocytes from normal and tumor-bearing C57BL/6 mice in 5 day cultures and measured proliferation by means of a standard 4-hour 3H thymidine incorporation assay. Potent inhibition of IL-2-induced proliferation (usually greater than 90%) resulted from high concentrations of IFN (100 to 1000 U/ml). Splenocytes from both tumor-bearing and normal mice, cultured alone or with mitomycin-treated syngeneic tumor, MCA106, exhibited similar patterns of inhibition. Unexpectedly, allostimulation with DBA/2 (H-2d) stimulators and C57BL/6 (H-2b) responders showed enhanced proliferation, most pronounced at lower IFN concentrations. Time course of IFN action revealed minimal duration of exposure necessary for inhibition to be less than 24 hours. Microfluorometric cell analysis showed the striking increase in Thy-1+ cells (55% to 96%) induced by IL-2 with parallel increase in Lyt2+ cells but almost complete disappearance of L3T4+ cells. IFN reduced IL-2-induced increase in Lyt-2+ cells (61% to 31%). These results emphasize the complexity of IL-2 and IFN interaction and the potential for dose-related antagonistic effects. This may be important in the clinical use of this combination. PMID- 2969633 TI - Transcoronary platelet activation and consumption in coronary artery disease: studies at rest. AB - The platelet count(PC), plasma platelet factor 4 (PF4) and plasma beta thromboglobulin(beta TG) have been measured in blood obtained from a peripheral vein, the aortic root and the coronary sinus in 7 patients with normal coronary arteries, 9 patients with lesser degrees of coronary artery disease(CAD) and in 13 patients with severe CAD under resting conditions. In each patient group values obtained in the peripheral venous blood were similar to those obtained in normal subjects. In each group values obtained in blood from the coronary sinus were similar to those obtained in blood from the coronary aortic root and in most instances these were similar to values obtained in peripheral venous blood. for example, in the 13 subjects with hemodynamically significant 3-vessel or 2-vessel CAD the mean values in blood from a peripheral vein, the aorta and the coronary sinus respectively were: PC-194, 205, and 208 x 10(9)/1; PF4-3.3, 3.7, and 3.5 ng/ml; and beta TG-15.5, 23.0 and 18.6 ng/ml. These findings provide no support for the occurrence of continuous platelet activation or platelet consumption in the coronary vessels or elsewhere in patients with stable CAD, under resting conditions, regardless of its severity. PMID- 2969634 TI - The simultaneous measurement of total and free protein S by ELISA. AB - An ELISA for the simultaneous measurement of total and free protein S is described. Polyethylene glycol precipitation was used to remove the C4b -binding protein/protein S complex from plasma. This allows free protein S to be measured. The total protein S was measured directly on plasma samples that had been diluted to dissociate the C4b binding protein/protein S complex. The concentration of total and free protein S found in normal subjects (n = 24) was 92.9 (SD = 10) u/dl and 40.3 (SD = 8.8) u/dl respectively. The within assay coefficient of variation was 5.1%. The between assay coefficient of variation was 5.7%. Warfarin therapy caused a reduction in the level of both total and free protein S but this reduction correlated poorly with the INR value. PMID- 2969635 TI - Alkyltin inhibition of ATPase activities in tissue homogenates and subcellular fractions from adult and neonatal rats. AB - Inhibition of ATPase activities by triethyltin (TET), diethyltin (DET), monoethyltin (MET), and trimethyltin (TMT) was studied in homogenates of brain and liver from adult and neonatal rats. In the adult, sensitivities were as follows: mitochondrial ATPase of liver much greater than Na+, K+-ATPase of brain approximately equal to mitochondrial ATPase of brain greater than nonspecific ATPase of brain and liver. MET did not produce significant inhibition. ATPase activities in brain and liver homogenates from TET-treated adult rats did not differ from controls. Mitochondrial ATPase in brain homogenates from 5-day-old rats was two orders of magnitude more sensitive to TET than brain homogenates from adult rats (IC50 of 2.5 microM in the 5-day-old neonate vs 260 microM in the adult). By contrast, isolated mitochondria and synaptosomal fractions from adult and neonatal brains were equally sensitive to TET (IC50 = 1-3 microM). At 10 days of age, following the onset of myelination, the IC50 for TET inhibition of brain mitochondrial ATPase increased to 71 microM. Myelin added directly to isolated mitochondria also reduced TET-induced inhibition. It is concluded that in vivo brain tin concentrations in 5-day-old rats following a neurotoxic dose of TET are sufficient to inhibit brain mitochondrial ATPase, whereas in adults, tin concentrations are insufficient for inhibition. In the adult rat, TET binding to myelin appears to prevent inhibition of brain mitochondrial ATPase, and the target of toxic action may be myelin. In the neonateal rat, TET may inhibit oxidative phosphorylation in unmyelinated brain tissue, leading to neuronal cell death. PMID- 2969636 TI - Electrocardiogram in primary pulmonary hypertension--with special reference to prognosis. AB - The electrocardiogram (ECG) in patients with primary pulmonary hypertension who died suddenly (Group II, 12 patients) and those who died of right heart failure (Group III, 19 patients) were compared with those of long-term survivors (Group I, 10 patients). In P waves, AP had a tendency to deviate more to the right in Groups II and III than in Group I. P terminal force in lead V1 tended to show a greater absolute value, and the amplitude of P waves in lead II were higher in Groups II and III than in Group I. The AQRS in Groups II and III deviated significantly much more to the right than in Group I. In lead V1, S waves had a tendency to be deeper and the R/S ratio greater in Groups II and III than in Group I. In leads V5 and V6, R waves and the R/S ratio significantly lower in Groups II and III than in Group I. S waves in lead V6 tended to be deeper in Groups II and III than in Group I. In the ECG patterns, S1O3, inverted T waves in leads II, III and a VF and qR patterns in lead V1 tended to appear more often in Groups II and III than in Group I. In Groups II and III, there was T wave inversion beyond V3 to the left chest leads and significant differences between Groups II and III, and Group I.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969637 TI - Specificity of the rearrangements of the T-cell receptor gamma gene in human lymphomas. AB - The structure and function of the human T-cell rearranging gamma gene are not completely understood. Several reports have suggested that this gene rearranges specifically in normal T cells, but the pattern of rearrangement in human lymphoid neoplasms is not clear. Some authors have described the rearrangements of this gene in unmanipulated leukemias as relatively specific for T-derived tumors, whereas others were unable to observe such specificity in malignant lymphomas. The present paper reports the analysis of the structure of the gamma gene in 32 lymphoid samples of different origin, with emphasis on non-T lymphomas. Four out of four T-cell lymphomas had this gene rearranged, whereas none of the 17 cases of B-cell lymphomas, 5 of Hodgkin's disease or 6 of nonneoplastic lesions showed any alterations of the gamma gene. Therefore, our data support the relative specificity of the gamma gene rearrangements in human T cell malignant proliferations. PMID- 2969638 TI - Psychologic aspects of patients participating in a phase I study with the synthetic retinoid 4-hydroxyphenyl retinamide. AB - One hundred and one patients participating in a phase I study with the synthetic retinoid 4-HPR (4-hydroxyphenyl retinamide) were evaluated. The study was set up by Veronesi et al. during 1986 at the National Cancer Institute of Milan. The patients were randomized into 4 groups of therapy: 25 in the placebo group, 25 in the group receiving a daily dose of 100 mg of HPR, 26 in the group receiving 200 mg/day of HPR, and 25 in the group receiving 300 mg/day of HPR. All patients were previously treated at our Institute for breast cancer. None had received adjuvant therapy, chemotherapy or hormone therapy. After 4-5 months from the beginning of treatment, all patients received a series of tests to evaluate anxiety, depression and sexual life. Moreover, during one the follow-up checkups after 4-5 months, the patients filled-out a self-scoring mood questionnaire. The results did not show any particular differences between the groups, although we found that the administered drug and experimental setting do not interfere with the psychologic state of the participating patients. PMID- 2969640 TI - [The role of coronary angioplasty at the time of initial coronary angiography in the treatment of unstable angina]. PMID- 2969639 TI - Alteration of opioid peptide circadian rhythm in cancer patients. AB - Endogenous opioid peptides have been seen to play a role in regulating immunity and tumor growth. This study was carried out to investigate opioid activity in human cancer. We evaluated by radioimmunoassay beta-endorphin plasma levels on blood samples collected at 9.00 a.m. from 121 cancer patients and 42 healthy subjects. In 22 cancer patients and in 12 controls, beta-endorphin circadian rhythm was also investigated. Finally, in 14 cancer patients and in 10 controls GH, PRL, FSH, LH and cortisol serum levels were measured after the administration of a metenkephalin analogue, FK 33-824 (0.3 mg i.v.). No significant differences were seen in beta-endorphin mean levels between cancer patients and normal subjects. Moreover, no differences were found between patients with or without metastases, nor between those with or without chronic pain. beta-Endorphin circadian rhythm appeared to be altered in 16/22 cancer patients, and anomalous hormonal responses to FK 33-824 were seen in 13/14 patients. This study shows an altered opioid activity in human neoplasms, whose clinical significance remains to be determined. PMID- 2969641 TI - Can combined DES and LHRH depot therapy (ICI 118630) prevent endocrinologic and clinical flare-up in metastatic prostate cancer? AB - Medical castration obtained with luteinizing hormone releasing hormone (LHRH) analogues in patients with prostate cancer is now well established. To block the initial stimulation of testosterone production and prevent the risk of the so called flare-up with this medication, we investigated short-term combination therapy with 1 mg of diethylstilbestrol (DES). Fourteen previously untreated patients with histologically proved metastatic prostatic carcinoma were treated with 1 mg DES po daily one week prior to the initiation of therapy with LHRH analogues depot injection of Zoladex (ICI 116630) and continued during four weeks after the first depot injection. LHRH depot form was maintained as long as patients experienced clinical benefit. Endocrinologic results show that in spite of 1 mg of DES a significant increase of testosterone is still observed in the first week after injection of the LHRH depot form. Hence, this combination is not useful to prevent endocrinologic and clinical flare-up in patients with prostate cancer treated with LHRH analogues. PMID- 2969642 TI - [A method of adapting the motor activity to the hot climate at the sanatorium stage of rehabilitation of patients after myocardial infarction]. PMID- 2969643 TI - [A portable aero-ionometer]. PMID- 2969644 TI - [Effect of oral intake of Essentuki mineral water No. 4 on gastric stump emptying in patients with dumping syndrome after gastrectomy]. PMID- 2969645 TI - [Effect of various concentrations of nitrosodimethylamine on the functional state of enzyme systems in rat liver mitochondria depending upon the duration of exposure]. AB - Activities of glutamate-, succinate-, malate dehydrogenases and H+-ATPase were studied in liver tissue and these of malate- and glutamate dehydrogenases--in blood serum of rats which received nitrosodimethylamine (0.1, 1.0 and 10 mg per I l of water) within 2.5 and 10 months. Cyclic alterations in activities of malate- and succinate dehydrogenases were found in liver tissue of these rats, while the activities of glutamate dehydrogenase and H+-ATPase were changed depending on concentration and duration of the carcinogen effect. Unfavourable alterations in the functional state of mitochondrial enzymatic systems were increased according to intensity and duration of nitrosodimethylamine effect. PMID- 2969646 TI - Effect of proximal gastric vagotomy on human gastric blood flow evaluated by endoscopic laser Doppler flowmetry. PMID- 2969647 TI - Microbial cleavage of zearalenone. AB - 1. Zearalenone, a fungal oestrogenic compound, was subjected to microbial transformation studies. Preliminary screening with 150 fungal species showed that Gliocladium roseum was capable of metabolizing zearalenone in 80-90% yields. 2. Large-scale fermentation with G. roseum produced a 1:1 mixture of 1-(3,5 dihydroxyphenyl)-10'-hydroxy-1-undecen-6'-one and 1-(3,5-dihydroxyphenyl)-6' hydroxy-1-undecen-10'-one. The compounds were isolated and purified at -20 degrees C, and identified using spectroscopic analysis and by comparison to products obtained from alkaline hydrolysis of zearalenone. PMID- 2969648 TI - Sulphation and glucuronidation of xamoterol in the dog: dose dependence and site of sulphation. AB - 1. Xamoterol has been administered both intravenously and orally over a 100-fold dose range to male beagle dogs. 2. Over the dose range examined, sulphation was not saturable, with the proportion of the dose excreted as the sulphate conjugate remaining constant. 3. Extensive first-pass sulphation of an oral dose of xamoterol occurred in the intestine with approximately 50% of sulphation occurring during absorption. 4. The intestine is not a major site of sulphation for circulating xamoterol. 5. The liver is not believed to play an important role in the first-pass sulphation of xamoterol. PMID- 2969649 TI - [Bronchoalveolar lavage in patients with sarcoidosis--methodologic aspects of the collection, immunologic characterization of lymphocytes and studies of immunoglobulin content]. AB - The activity of alveolitis in 25 patients with pulmonary sarcoidosis was determined by analysis of T-Lymphocytes and their subpopulations collected by broncho-alveolar lavage. The characterization of the lymphocytes was performed with monoclonal antibodies (BL-series). An amount of T-lymphocytes higher than 28% of all cells and a proportion of T4/T8-lymphocytes of 8:1 and over are signs of the activity of the alveolitis and an indication for corticoid therapy. PMID- 2969650 TI - [Hematoporphyrin derivative-induced tumor fluorescence]. AB - Detection of HpD-sensitized tumour was successful by use of HBO 200 mercury lamp, Argon ion laser, as well as for N2-pumped dye laser. After 20 hours period of HpD application the localization of the Ehrlich ascites tumour in mice was identified macroscopically and by spectroscopy. To exclude red emissions of detection sources the use of dye lasers is recommend. PMID- 2969651 TI - Influence of microbiological isolation on some microbial intestinal parameters. PMID- 2969652 TI - [Closed traumatic rupture of the abdominal wall]. PMID- 2969653 TI - [Correlation of maternal androgens in early and late pregnancy. A correlation with fetal size and sex]. AB - In the course of prospective investigations blood was withdrawn for steroid analysis in 50 gravidae between 12th and 16th and between 26th and 30th week of gestation. The levels of androstenedione, testosterone, dehydroepiandrosterone sulfate, 17-hydroxyprogesterone and sex hormone binding globuline were determined by radioimmuno-assay evaluating a possible correlation between the androgen values and growth retardation or the sex of the fetus. Neither during the early nor during the late period of pregnancy significant differences concerning the androgen levels could be detected among both sexes. Worth mentioning is the significant decrease of the DHEA-S levels during the late period of gestation, which occurred in both groups, whereas the androstenedione, testosterone and especially the 17-OH-P levels showed a significant increase. The SHBG values almost did not change at all. PMID- 2969654 TI - Pulsed Doppler echocardiography in the diagnosis and evaluation of aortic regurgitation. PMID- 2969655 TI - Abnormal response to handgrip in hypertensive patients with increased left ventricular mass, assessed by echocardiography. PMID- 2969656 TI - Noninvasive assessment of the functional condition of the left ventricle in essential hypertension. PMID- 2969657 TI - Differential binding of 125I-IGF-I preparations to human fibroblast monolayers. AB - Specific, high affinity binding of 125I-IGF-I to the type I IGF receptor on human fibroblast monolayers was not altered by varying feeding schedules, serum lots, washing procedures, or incubation times and temperatures. However, markedly different competitive binding curves were obtained when different iodinated IGF-I preparations were used. Five of six radioligands bound preferentially to the type I IGF receptor on human fibroblast monolayers, with 50% displacement at 4-8 micrograms/l unlabelled IGF-I; with one radioligand a paradoxical 20-200% increase in 125I-IGF-I binding was observed at low concentrations of unlabelled IGF-I, while concentrations as high as 100 micrograms/l IGF-I failed to displace this radioligand. The latter binding pattern cannot be accounted for by 125I-IGF I binding to the type II IGF receptor. These data indicate that various radioligands may have preferential affinities for different IGF-I binding sites on human fibroblast monolayers. PMID- 2969658 TI - Is perlaparoscopic bacteriological analysis sufficient for establishing the etiology of P.I.D.? AB - The Authors have studied the usefulness and the limits of laparoscopy in the etiologic diagnosis of P.I.D.; for this purpose 21 patients affected with P.I.D. underwent microbiological sampling from cervix and/or vagina and from Fallopian tube during laparoscopy. The findings have shown a discrepancy between the microbiological isolation from vagina/endocervix and from salpinx and remarkably the loss of evidence of S.T.D. agents from tuba with, conversely, frequent isolation of these agents (34.7%) from cervical swabs. Therefore, the Authors believe that the percelioscopic microbiologic study of endosalpinx is essential for the correct etiologic diagnosis of P.I.D., especially in anaerobic germ infections; yet the technical problems of percelioscopic samples and the biological features of S.T.D. agents require the simultaneous bacteriological sampling from the cervix and/or vagina. PMID- 2969660 TI - Echography in obstetrics and gynecological emergencies: the ectopic pregnancy. AB - Sonographic findings suggestive of ectopic pregnancy in 23 patients are reported by the Authors. In accordance with the data from the literature the real time sonography has shown in this study a diagnostic accuracy of 76%. PMID- 2969659 TI - Laparoscopy in obstetrics and gynecological emergencies. AB - The Authors underline the importance of the laparoscopy in the Obstetrical and Gyneacological emergencies. For this purpose, 74 consecutive laparoscopies, performed as emergencies at the Department of Obstetrics and Gynaecology of the University of Pavia, are considered and analyzed. The Authors stress that the celioscopy has allowed a precise diagnosis in all the cases examined, and, even thanks to percelioscopic surgery, has avoided some useless laparotomies. Even more, when it was necessary to perform a major surgical act, the right laparoscopic diagnosis has addressed the therapy in several cases to conservative surgery. PMID- 2969661 TI - Hysteroscopic metroplasty: the role of ultrasound in the diagnosis and monitoring of patients with uterine septa. AB - 52 patients with septate uterus underwent hysteroscopic metroplasty in the Obstetric and Gynecologic Clinic "R" in Palermo. In this group of patients ultrasound examinations were also performed in order to define their role in diagnosis of type of malformations and in monitoring the endoscopic surgical procedure. Preliminary results seem to indicate that real-time ultrasound is a reliable technique for both purposes. PMID- 2969662 TI - [Perihepatitis due to Chlamydia trachomatis. The diagnostic and therapeutic value of laparoscopy]. PMID- 2969663 TI - Vasorelaxant and natriuretic activities of synthetic atrial peptides. AB - The vasorelaxant and natriuretic activities of synthetic atrial natriuretic peptides (ANF) were compared by measuring relaxation of histamine-contracted rabbit aortic rings and changes in urinary sodium excretion in anesthetized rats. The peptides studied and their relation to pro-ANF were: atriopeptins (AP) I, II and III (ANF 103-123, 103-125, 103-126), cardionatrin (CN) (ANF 99-126), alpha human atrial natriuretic polypeptide (alpha-hANP) (human 99-126) and a fragment of the last (hFrag) (human ANF 105-126). The concentration causing 50% relaxation (EC50) in the vasorelaxant assay for API was greater than that of the other peptides (p less than 0.05). The EC50 for hFrag was 20 times greater than that of alpha-hANP, but this difference was not statistically significant. API caused less natriuresis compared with the other peptides (p less than 0.05). The natriuresis induced by CN was significantly greater than that of the other peptides at the 485 pmol dose. These results indicated that, except for API, the peptides studied has similar vasorelaxant and natriuretic activities. PMID- 2969664 TI - Characterization of interleukin 1 receptors in brain. PMID- 2969665 TI - [What can be expected of intracavernous injections?]. PMID- 2969666 TI - Percutaneous transluminal angioplasty prior to carotid cavernous fistula embolization. PMID- 2969667 TI - Myocardial protection by perfluorochemical infusion during transient ischemia produced by balloon coronary occlusion. AB - To assess the efficacy of the perfluorochemical Fluosol-DA 20% for myocardial protection during repeated periods of balloon occlusion of the left circumflex coronary artery, 25 anesthetized dogs were randomized to receive either oxygenated perfluorochemical, (Fluosol-DA [F]; n = 10) or oxygenated Ringer's lactate (R; n = 6), at the rate of 30 ml/min, during inflation of the balloon. A control group (C; n = 9) received no infusion. A total of eight inflations were performed, each lasting 90 seconds, followed by an equivalent deflation time. Hemodynamics, ECGs, regional myocardial function, and biochemical parameters were studied. Significant differences were noted in ST segment elevation at 90 seconds of inflation in the F (1.5 mm +/- 0.6), C (3.7 mm +/- 0.75), and R (2.9 mm +/- 0.75) groups (F vs C or R p less than 0.05). This was associated with significant improvement in radial shortening in the jeopardized zone at 45 seconds into occlusion in the F group compared to the C and R groups (F = 21.1% +/- 5.1 vs C = 3.5% +/- 4.5 or R = 1.1% +/- 3.2; p less than 0.05). Results of electron microscopy showed reversible changes of ischemia within the mitochondria, and these were most marked in the C and R groups compared to the F group. Endothelial swelling was mild and was present only focally in the R and C groups. Thus perfluorochemicals may enhance the safety and efficacy of balloon angioplasty. PMID- 2969668 TI - Atrial natriuretic factor release is enhanced by incremental atrial pacing. AB - There is ample evidence from animal models indicating that secretion of atrial natriuretic factor (ANF) can be induced by an increase in atrial contraction frequency or atrial distension. The influence of these stimulatory signals on ANF secretion in humans has not been fully elucidated. We assessed the responses to graded right atrial pacing in 28 patients (aged 33 to 70 years) at rates of 100, 125, and 150 beats/min on right atrial pressure, left atrial size (by two dimensional echocardiography in 9 of 28 patients), and circulating plasma ANF levels. At pacing rates of 125 and 150 beats/min, ANF levels increased from a baseline value of 64 +/- 9 fmol/ml (mean +/- SEM) to 89 +/- 13 fmol/ml (pp less than 0.05) and to 132 +/- 17 fmol/ml, respectively (p less than 0.001). Right atrial pressure increased from a baseline value of 4.1 +/- 0.7 mm Hg to 4.5 +/- 0.6 mm Hg at a pacing rate of 125 beats/min (p less than 0.05) and to 6.1 +/- 0.8 mm Hg at a pacing rate of 150 beats/min (p less than 0.001). Left atrial dimension increased from a baseline value of 44.5 +/- 3 mm to 49.5 +/- 3 mm at a pacing rate of 125 beats/min (p less than 0.05) and increased further to 52.5 +/- 3 mm at a pacing rate of 150 beats/min (p less than 0.001). No significant changes in atrial pressure or size or in plasma ANF were observed at a pacing rate of 100 beats/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969669 TI - Antimitochondrial autoantibodies in myocardial hypertrophy: comparison between hypertrophic cardiomyopathy, hypertensive heart disease, and athlete's heart. AB - Antimitochondrial autoantibodies (AMA) were tested by indirect immunofluorescence in three groups of subjects with different types of myocardial hypertrophy: 35 patients affected with hypertrophic cardiomyopathy (HC), 20 patients with cardiac hypertrophy secondary to essential hypertension, and 35 active endurance athletes with exercise-induced left ventricular hypertrophy. Forty-two healthy subjects served as a control group. Left ventricular hypertrophy was considered a left ventricular mass (LVM) echocardiographically calculated (Devereux formula), exceeding 244 gm or a LVM index exceeding 122 gm/m2 (greater than 2 SD from a previously studied normal population). AMA were found in 15 of 35 (43%) patients with HC and in 6 of 20 (30%) patients with hypertensive heart disease (p less than 0.01); in contrast, AMA were not present in the sera of athletes or in the sera of controls. Although the significance of AMA in subjects with pathologic myocardial hypertrophy has not yet been established, their absence in the sera of athletes strengthens the opinion that cellular changes, as a compensatory response of the myocardium to a work overload, have a physiologic fashion in these cases. Moreover, identification of AMA in the sera of athletes with disproportionate severe left ventricular hypertrophy of uncertain origin may be helpful to ensure a single diagnosis. PMID- 2969670 TI - Alterations in myocardial thallium-201 distribution in patients with chronic systemic hypertension undergoing single-photon emission computed tomography. AB - To characterize thallium-201 distribution in single-photon emission computed tomography (SPECT) cardiac images and polar bullseye maps, 100 patients with chronic systemic hypertension due to end-stage renal disease were studied and the results compared with those in 35 normotensive control subjects. Thallium-201 SPECT was performed after exercise in all control subjects and 70 hypertensive patients, and after intravenous dipyridamole in 30 patients. A frequent finding in hypertensive patients was a fixed decrease in the normal lateral-to-septal count density ratio in immediate thallium-201 SPECT images (1.02 +/- 0.10 vs 1.17 +/- 0.08 in control subjects, p less than 0.00001) and in 3-hour delayed images (1.02 +/- 0.11 vs 1.11 +/- 0.08 in control subjects, p less than 0.00001). No significant difference in count density ratio was present in patients undergoing treadmill versus diypridamole intervention. In 35 patients the count density ratio was greater than 2.0 standard deviations below the normal mean, creating the false impression of a fixed lateral defect (i.e., myocardial infarction). In 12 patients, myocardial wall thickness was measured at end-diastole by 2 dimensional echocardiography. Wall thickness was increased (greater than 11 mm) in all patients. The mean lateral-to-septal wall thickness ratio was 1.08 +/- 1.11; in no patient was the ratio less than 0.76 to indicate selective septal hypertrophy. The lateral-to-septal wall thickness and lateral-to-septal thallium 201 count density ratios correlated poorly (r = 0.43).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969671 TI - First dose response and 24-hour antihypertensive efficacy of the new once-daily angiotensin converting enzyme inhibitor, ramipril. AB - The reduction in blood pressure (BP) after the first dose and after 8 weeks of treatment with a new once-daily angiotensin converting enzyme (ACE) inhibitor, ramipril, was examined in 12 untreated hypertensive patients, using ambulatory intraarterial BP monitoring. The first period of monitoring began 24 hours before the first dose was given, and continued for 24 hours afterwards. A second 24-hour period of monitoring was carried out after 8 weeks of treatment, commencing immediately after the morning dose. Angiotensin II levels and serum drug levels were measured at 0, 2, 6 and 24 hours after the acute dose. BP decreased progressively from the first hour after the first dose, reached a maximum in the fifth hour (p less than 0.001) and then the effect diminished. The maximum reduction of systolic BP in any patient was 64 mm Hg, the minimum 4 mm Hg. Blood pressure was significantly (p less than 0.05) reduced throughout the 24 hours after dosing, with a mean daytime reduction of 13/12 mm Hg, and a mean nighttime reduction of 15/7 mm Hg. Angiotensin II levels were significantly (p less than 0.02) and maximally reduced by 2 hours after administration, but the reduction was no longer significant after 24 hours. Serum drug levels were also maximal 2 hours after administration. The trial population could be clearly divided into groups of good and poor responders on the basis of BP reduction. The angiotensin II levels were higher before treatment, and decreased further, in all patients with a good response than in those with a poor response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969672 TI - Relation of concentric left ventricular hypertrophy and extracardiac target organ damage to supranormal left ventricular performance in established essential hypertension. AB - Increased cardiac performance has been documented in patients with early systemic hypertension, but its prevalence and determinants in patients with uncomplicated sustained essential hypertension have not been characterized. Radionuclide cineangiography in 116 patients with uncomplicated essential hypertension showed that 12 of 116 (10%) had supranormal resting left ventricular (LV) ejection fraction (greater than 70%, above the highest value in normal subjects), while 104 patients had a normal resting ejection fraction (45 to 70%). Patients with a high resting ejection fraction had higher systolic and diastolic blood pressure compared with patients with normal resting ejection fraction (182 mm Hg vs 169, p less than 0.01, and 110 vs 103, p less than 0.05, respectively), markedly greater echocardiographic LV mass (136 vs 94 g/m2, p less than 0.01), smaller ventricular dimensions in systole (2.5 vs 3.1, p less than 0.01) and diastole (4.4 vs 4.9, p less than 0.05), and higher relative wall thickness (0.61 +/- 0.20 vs 0.39 +/- 0.98, p less than 0.001). Patients with supranormal resting ventricular performance had lower end-systolic wall stress than normal volunteers or patients with normal resting LV function (48 vs 64 vs 74 X 10(3) dynes/cm2, respectively). Patients with an elevated LV ejection fraction also had significantly more abnormal funduscopic examinations and greater proteinuria. Thus, a subset of essential hypertensive patients with moderately to severely elevated blood pressure developed marked concentric LV hypertrophy associated with subnormal end systolic stress and supranormal LV performance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969673 TI - Left ventricular mass as determined by magnetic resonance imaging in male endurance athletes. AB - Although many studies of the effect of dynamic exercise training on left ventricular (LV) mass have been reported, controversy continues to exist. Previous work has been criticized because of the techniques used for measuring LV mass, the variable level of training of the subjects recruited and the methods used to normalize the data. In an attempt to resolve this controversy, LV mass was determined using the very accurate and reproducible technique of magnetic resonance imaging (MRI). Highly trained competitive athletes including cross country skiers, endurance cyclists and long distance runners (VO2max = 77 +/- 1, 72 +/- 2 and 75 +/- 2 ml (kg X min)-1, respectively) were examined. The data were normalized for body weight, body surface area and lean body mass. LV mass was significantly greater in skiers (239 +/- 9 g), runners (244 +/- 10 g) and cyclists (258 +/- 11 g) when compared with nonathletic control subjects (189 +/- 6 g) (p less than 0.001), which represents percent differences of 26, 29 and 37%, respectively. LV mass remained greater in the athletes, regardless of the method used to normalize the data. In addition, there was a good correlation between LV mass and VO2max (r = 0.80, p less than 0.001). It was concluded that LV mass is significantly greater in highly trained competitive endurance athletes and that normalizing LV mass with respect to body weight, body surface area or lean body mass does not alter this relation. PMID- 2969674 TI - Evaluation of a QRS scoring system for estimating myocardial infarct size. VII: Specificity in a control group with right ventricular hypertrophy due to mitral stenosis. PMID- 2969675 TI - Immunopharmacodynamic profiles in children with renal allografts receiving cyclosporine therapy. AB - Eleven renal transplant patients between the ages of 4 and 19 years, on a single daily oral dose of cyclosporine (CsA) and either a low oral dose or no dose of prednisone, had venous blood drawn at periodic intervals throughout a 24-hour period. CsA levels were measured by whole blood radioimmunoassay. All the patients had similar patterns of CsA pharmacokinetics with a single peak blood level at two to eight hours after the drug was given. Plasmas separated from the bloods at 37 degrees C were added to third party mixed lymphocyte reactions (MLR). The kinetics of suppression of the MLR by serial plasmas did not follow the CsA levels. Instead, we observed patterns of suppression similar to those that have been described in adults. Five patients had pattern I with two peaks of plasma-mediated MLR suppression, and had no rejection episodes. Two of the patients had pattern II with only one peak of MLR suppression, and both had episodes of acute rejection. One patient showed pattern III with a pleateau of MLR suppression, and has had no rejection episodes and no obvious CsA toxicity. Three patients showed pattern IV with a continuously low level of plasma-mediated MLR suppression throughout the day, and two of them have had severe rejection episodes. Immunopharmacodynamic profiling (IP) may prove to be useful in individualizing therapeutic regimens for patients with renal allografts treated with CsA. PMID- 2969676 TI - Is there a role of chromosome 1 in the clinical expression of diabetes mellitus? PMID- 2969677 TI - The differential response in atrial natriuretic peptide release during exercise in patients with and without ischemic heart disease. AB - The elevation of cardiac filling pressure induces the release of atrial natriuretic peptide into the circulation. Ischemia during exercise in patients with coronary artery disease may manifest itself with elevation of cardiac filling pressure before the onset of electrocardiographic changes or chest pain. Thus, patients with ischemic heart disease might have an elevated circulating atrial natriuretic peptide after exercise. The present study investigated the effect of exercise on circulating atrial natriuretic peptide in patients with and without ischemic heart diseases. Group 1 was composed of five patients who had ischemic heart disease by clinical history, previous myocardial infarction, angina or angiographically proven coronary artery disease and positive electrocardiogram during exercise. Group 2 was composed of five patients without ischemic heart disease and negative electrocardiogram response. Heart rate, blood pressure, and atrial natriuretic peptide were measured during routine treadmill exercise testing using the Bruce protocol. Our results indicate that the rate of rise of heart rate (12.3 +/- 1.8 vs. 8.5 +/- 0.7 beats/min/min), blood pressure (7.1 +/- 1 vs. 4.2 +/- 0.8 mm Hg/minute), and atrial natriuretic peptide (4.1 +/- 1 vs. 1.4 +/- 0.3 pg/ml/min) was significantly elevated in patients with ischemic heart disease compared to the group 2 patients. These findings suggest that the disproportionate elevation of atrial natriuretic peptide after exercise in ischemia may be caused by elevation of cardiac filling pressure, which may provide a noninvasive method for the diagnosis of ischemic heart disease. PMID- 2969678 TI - Cough caused by cilazapril. AB - Angiotensin-converting enzyme (ACE) inhibitors are a group of drugs recently introduced to treat hypertension and congestive heart failure. There are many reports of a dry cough in patients treated with (ACE) inhibitors, but this is often considered a rare side effect. Eleven of 30 patients treated with the investigational ACE inhibitor cilazapril complained about a chronic cough. PMID- 2969679 TI - Ipratropium bromide in COPD and asthma. PMID- 2969680 TI - Maternal serum hexosaminidase A in pregnancy: effects of gestational age and fetal genotype. AB - The diagnostic usefulness of sulfated fluorogenic substrates in carrier detection of Tay-Sachs disease in serum during pregnancy was assessed by testing coded samples. Gradual increase in serum hexosaminidase activities toward these substrates was observed throughout pregnancy in both carrier and non-carriers of the Tay-Sachs gene, but absolute discrimination between the 2 genotypes could not be achieved even when values were compared within the same gestational age. Examination of isolated isozyme fractions with the sulfated substrates showed that the increased activities during pregnancy were due to a genuine increase in hexosaminidase A and not associated with the elevation of hexosaminidase I (or P), which was evident only with unsulfated substrates. The extent of the increase was influenced by the genotype of the fetus as indicated by higher values in pregnant carriers who carried non-carrier fetuses. We conclude that determination of serum hexosaminidase A during pregnancy by sulfated fluorogenic substrates may have a prenatal diagnostic value when used in obligate heterozygotes for Tay Sachs disease, but is unreliable for screening purposes. PMID- 2969681 TI - Maternal age-specific rates for Down syndrome: changes over time. AB - There have been conflicting reports about whether incidence rates at given maternal ages for Down syndrome (DS) are changing over time. British Columbia provides an excellent opportunity to look at this question as there is 1) nearly complete ascertainment of DS livebirths, 2) a relatively large number of annual births, 3) data on a 20-year time period, 4) data on age-specific fertility rates over this period, and 5) cytogenetic data on all pregnancies terminated through the provincial prenatal diagnosis program since its inception. Data on 856 liveborn DS infants over a 20-year period (1964-1983) were analyzed. Controlling for maternal ages, the data were examined for transient changes in incidence over time and for linear and cyclic trends. No significant (P greater than 0.05) changes in DS incidence were observed over this 20-year period in any maternal age group. PMID- 2969682 TI - Down syndrome and trisomy 18 in the Bedouins. PMID- 2969683 TI - Maternal age in Down syndrome. PMID- 2969684 TI - Abnormal scleral findings in uveal effusion syndrome. AB - We successfully treated a patient with uveal effusion syndrome and abnormal sclera with a partial-thickness sclerectomy. Part of the sclera was immediately cultured, and the excised sclera and the cultured cells were examined by electron microscopy. The sclera demonstrated increased glycosaminoglycan-like deposits between the scleral fibers. The cultured scleral cells showed large intracellular glycogen-like deposits, which were not seen in cells cultured from two control scleras. These findings may be the result of a metabolic defect, which causes a thick, impermeable sclera in some cases of uveal effusion. PMID- 2969685 TI - Psychotherapeutic implications of severe physical disability. AB - Some effects of physical disability on psychological functioning are reviewed. Special roles are proposed for fantasy and therapist counter-transference in psychotherapy with physically disabled people. This report of the treatment of a man with severe cerebral palsy has implications for the physical and psychological management of the disabled. PMID- 2969686 TI - The impact of a UNICEF-assisted rural water project on the prevalence of guinea worm disease in Asa, Kwara State, Nigeria. AB - This paper demonstrates that protected water supplies, in the form of boreholes, can reduce the prevalence of dracunculiasis (guinea worm disease) in affected communities from a point prevalence of greater than or equal to 50% to 0% or near 0% within 3 years of intervention. Studies in Nigeria show that boreholes sited within a village and used exclusively for drinking water are most effective, while less accessible or malfunctioning boreholes have a less dramatic impact on prevalence. In contrast to the situation in villages served with boreholes, the prevalence of guinea worm in the unserved villages remained almost unchanged. The rapid benefits of protected rural water supplies and the decline of dracunculiasis, including the rise in school enrollment and fall in the rate of school absenteeism, are highlighted together with other socioeconomic benefits. PMID- 2969687 TI - [Critical evaluation of invasive and noninvasive study methods used in the gynecology of childhood and adolescence]. PMID- 2969688 TI - [Diagnosis of tumors and tumor-like formations of the ovaries]. PMID- 2969689 TI - [The importance of laparoscopy in the diagnosis of tubal sterility]. PMID- 2969690 TI - Ultrastructural observations on muscle spindles in extraocular muscles of pig. AB - Human and some mammals such as the sheep, goat and domestic and wild pigs have more or less muscle spindles in the extrinsic eye muscles, especially the domestic pigs having abundant muscle spindles (Matsuyama, 1987). The muscle spindles play a large role in maintaining the stable visual posture of the eyeballs. To define the morphological properties of the muscle spindles relative to the eye movement, the ultrastructure of the spindles was investigated in 6 extraocular muscles of the pigs by electron microscopy. The muscle spindles in the pig extraocular muscles consist of 4 to 5 intrafusal muscle fibers, one of which is nuclear bag fiber and 3 to 4 are nuclear chain fibers. The outer capsule is thin, composing of few layers, and the inner capsule ramifying to enwrap the individual fiber, accompanied by the medullated and unmedullated nerve fibers and blood capillaries. The nuclear bag fiber, 14 micron in diameter, is innervated by the atypical annulospiral sensory terminals and the chain fiber by the typical annulospiral terminal packed with mitochondria and microvesicles. The intrafusal fibers are innervated by the flower-spray sensory terminals anchoring deeply into the sarcoplasma, having abundant neurotubules and few mitochondria. The gamma motor end-plates have a relatively smooth synaptic cleft with a width of 70 nm and synaptic boutons containing few synaptic vesicles, sometimes, revealing a shallow fold of postsynaptic sarcolemma and abundant synaptic vesicles. The alpha motor end-plates reveal a relatively smooth synaptic cleft with a width of 80 nm, sometimes with a rough postsynaptic infolding, and boutons containing few synaptic vesicles and small-sized mitochondria. The satellite cells are innervated by the sensory terminals in various ways. The muscle spindles in the pig extraocular muscles are found to be much simpler in structure than those in the other antigravity muscles of the body. Their ultrastructure seems to reflect the morphological adaptation relative to the eyebal movement. PMID- 2969691 TI - Effects of fine-grained diet on annulospiral endings of muscle spindles in the masseter muscle in developing and adult mice. AB - In the masticatory muscles, neuromuscular spindles have a very important role in controlling the jaw movement since they act as stretch receptors in skeletal muscles. The continuous intake of fine-grained diet which is easily chewable leads to degeneration of the sensory endings of Ia fibers in many muscle spindles of the mouse masseter muscle in only 120 d after birth. PMID- 2969692 TI - Comparison of clinical judgment, Doppler ultrasound, and fluorescein fluorescence as methods for predicting intestinal viability in the pony. AB - Strangulation obstruction was induced in anesthetized ponies for periods of 2 and 3 hours by clamping 45-cm segments of jejunum and associated veins (venous strangulation obstruction) and arteries and veins (arterial and venous strangulation obstruction). Four segments were studied in each of 7 ponies allowed to survive 12 hours, 2 segments in a pony that was allowed to survive 1 hour, and 1 segment in each of 10 ponies allowed to survive 42 days after the strangulation periods ended. Fifteen minutes after the periods of strangulation obstruction ended, the viability of test segments was assessed by clinical judgment (40 segments), fluorescein fluorescence (40 segments), and Doppler ultrasound (32 segments). Because the test segments were normal at necropsy in long-term survivors, all segments were designated as viable. The overall accuracy of the methods used to predict viability was 88% for Doppler ultrasound and 53% each for clinical judgment and fluorescein fluorescence (P less than 0.005). Failures in the last 2 techniques could be attributed to their tendency to score venous strangulation obstruction segments as nonviable (90% for each). Doppler ultrasound was 94% accurate in these segments. PMID- 2969693 TI - [Identification in Plasmodium falciparum of a protein specifically binding human fibronectins]. AB - A fibronectin binding protein (FnBp) was identified in 3H isoleucine labeled P. falciparum schizonts using affinity chromatography on human fibronectin (Fn) coupled to Sepharose 4B. After incubation of Nonidet-P 40 parasite lysate with Fn Sepharose, elution was performed with SDS-PAGE buffer. Analysis of FnBp by SDS PAGE demonstrated a major band which migrated with an apparent Mr of 70,000 under reducing conditions. This band was not found when human or rabbit IgG coupled Sepharose 4B were used instead of Fn as control. PMID- 2969694 TI - [Seborrheic dermatitis in infants: a clinical form of atopic dermatitis?]. PMID- 2969695 TI - Twofold CuZnSOD activity suggesting homozygous submicroscopic duplication of chromosome 21 in the orangutan. AB - A twofold CuZnSOD activity was observed in an orangutan as compared to man and a chimpanzee. This suggests that a submicroscopic duplication of chromosome 21 (similar to those demonstrated in patients with the trisomy 21 phenotype but with a normal karyotype) has occurred in the homozygous state during evolution of the orangutan phylum. Such duplications could be important evolutionary mechanisms, together with visible chromosome rearrangements and classical gene mutations. PMID- 2969696 TI - Dermatoglyphic patterns in senile dementia of Alzheimer's type. AB - Several reports suggest a genetic relationship between senile dementia of Alzheimer type (SDAT) and Down's syndrome. We have analyzed fingerprints and palmar patterns in an elderly female population comprising a group of 34 patients with probable SDAT, a group of 20 patients with other dementias, and a group of 20 non-demented controls. A bilateral Sydney line was found to be significantly more frequent in the SDAT group than in the two other groups (p less than 0.01, sensitivity 30%, specificity 95%, positive predictive value 91%, negative predictive value 61%). A bilateral Sydney line was as frequent in the SDAT group as in Down's syndrome. The limit value of the index of transversality equal or superior to 31, which is considered as a feature of Down's syndrome, was significantly more frequent in the SDAT group than in the two other groups (right hand p less than 0.05, left hand p less than 0.02). A bilateral discriminant value of this index was also significantly more frequent in the SDAT group than in the two other groups (p less than 0.02), as was an index of transversality higher than 31 on at least one hand (p less than 0.01). In contrast with other reports, we haven't found significantly different frequencies of digital ulnar loops and true hypothenar patterns between the SDAT group and the two others. PMID- 2969697 TI - Molecular cloning and sequence analysis of human placental ferredoxin. AB - We have characterized several clones specific for the human iron-sulfur protein, ferredoxin, which is involved in electron transfer to mitochondrial cytochromes P 450. Clones were isolated from a human placental cDNA expression library in lambda gt11 by immunoscreening with antibody to bovine adrenal ferredoxin. One clone contained the entire amino acid coding sequence (552 bp) together with 27 bp at the 5'-terminus and approximately 0.9 kb at the 3'-terminus; this form appears to correspond to the major mRNA species of approximately 1.7 kb observed on Northern blots of placental mRNA. The deduced amino acid sequence suggests that human ferredoxin is synthesized as a precursor of 184 amino acids (Mr 19,371) which is cleaved to yield a polypeptide of 124 amino acids (Mr 13,546). The mature protein is highly acidic, and the sequence is very similar to those of bovine and porcine adrenodoxins with the exception of substitutions and variations in length at the C-terminus. The N-terminal precursor segment, on the other hand, is considerably diverged from that determined for bovine adrenodoxin, but is similar in overall basicity and the pattern of occurrence of arginine residues. PMID- 2969698 TI - Influence of ligands on the aggregation of the normal and mutant forms of phosphofructokinase 2 of Escherichia coli. AB - The aggregation states of Escherichia coli phosphofructokinase 2 (Pfk-2) and of a mutant enzyme (Pfk-2*) altered in the inhibitory allosteric site for MgATP were measured in the presence and in the absence of substrates and products of the reaction. When sucrose gradient ultracentrifugation experiments were performed in the absence of added ligands, both enzymes sedimented as dimers. Likewise, at low concentrations of both substrates (0.1 mM) the aggregation state of Pfk-2 and Pfk 2* corresponded to a dimer. However, in the presence of 1 mM MgATP alone, Pfk-2 sedimented as a tetramer, whereas Pfk-2* sedimented as a dimer. At a low fructose 6-phosphate concentration (0.1 mM) and an inhibitory concentration of MgATP (4 mM), Pfk-2 sedimented as a tetramer. However, at the same MgATP concentration but at a higher fructose-6-P concentration (1 mM), a condition under which Pfk-2 is not inhibited by the Mg-nucleotide complex, the enzyme sedimented as a dimer. Pfk 2* is not inhibited under these conditions and sedimented as a dimer in each case. Thus, the effectiveness of MgATP in promoting the aggregation of Pfk-2 and Pfk-2* parallels the inhibitability of the enzymes by the nucleotide complex. However, ATP4-, a potent inhibitor of Pfk-2 and Pfk-2* that binds to the catalytic site of the enzymes, had no effect upon their aggregation states. Possibly Pfk-2* is not able to form a tetramer because of an alteration in the regulatory site for the Mg-nucleotide complex. PMID- 2969699 TI - Regulation of fatty acid oxidation in rat brain mitochondria: inhibition of high rates of palmitate oxidation by ADP. AB - Regulation of oxidation of [1-14C]palmitate in rat brain mitochondria has been investigated in purified mitochondria of nonsynaptic origin prepared by use of a Ficoll/sucrose density gradient. The mitochondrial preparation contained considerable Mg2+-ATPase activity, but was virtually free of contamination with nonmitochondrial fractions. Palmitate oxidation was inhibited by increasing the concentration of ATP in the assay system to near-physiological levels (2 mM), and the inhibition at 2 or 4 mM ATP was analyzed by comparing it with palmitate oxidation at near-maximal rates with low levels of ATP (0.5 or 1 mM). Inhibition was increased by the addition of ADP or by increasing the concentration of Mg2+ in the assay system, whereas inhibition was decreased by decreasing the concentration of mitochondrial protein or L-carnitine in the assay system. Increasing CoA concentration also had a deinhibitory effect. With 0.5 or 1 mM ATP, however, neither inhibition by added ADP nor protein concentration-dependent inhibition was observed, and the rate of oxidation was saturated with increasing concentrations of Mg2+, L-carnitine, or CoA. These results indicated that ADP was involved in the inhibition of high rates of palmitate oxidation in the presence of sufficient ATP and L-carnitine. The inhibitory effect of increasing the concentration of mitochondrial protein could be explained by the enhanced amounts of ADP present in the preparation; similarly, increased concentrations of Mg2+ would provide higher levels of ADP by stimulating the Mg2+-ATPase reaction. We discuss the possibility that the transport of ADP across the inner membrane of brain mitochondria is coupled to the inhibition of palmitate oxidation. PMID- 2969700 TI - Characterization of the peribacteroid membrane ATPase of lupin root nodules. AB - Peribacteroid membranes can be isolated in essentially pure form from 20-day lupin root nodules by osmotic shock of the purified membrane enclosed bacteroids. The ATPase (EC 3.6.1.3) associated with this membrane has an acid pH optimum (5.25) and is specific for ATP (Mg-ATP Km = 0.16 mM). The enzyme activity requires magnesium or manganese ions, is slightly stimulated by the cations potassium and rubidium, and is inhibited by vanadate, diethylstilbestrol, N,N' dicyclohexylcarbodiimide, fluoride, molybdate, and calcium. Molybdate and fluoride sensitivity do not in this case indicate the presence of significant nonspecific phosphatase activity. The ATPase is not inhibited by oligomycin, azide, or the soluble carbodiimide 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. In some respects the lupin peribacteroid membrane ATPase appears to differ from the plasma membrane ATPase of other plants. PMID- 2969701 TI - T-cell subset assay. A useful differentiating marker of atopic and seborrheic eczema in infancy? AB - To confirm T-cell changes previously reported in atopic eczema, to compare T-cell subset values in atopic and seborrheic infants, and to determine whether T-cell subset assay would be a useful differentiating marker between atopic and seborrheic infants, three age- and sex-matched groups of normal, seborrheic, and atopic infants were recruited. T-cell subset values, total serum IgE, and serum IgE to specific allergens (Dermatophagoides pteronyssimus, cat epithelium, dog dander, rye grass, egg white, and milk) were measured. The results showed that total and allergen-specific IgE levels were higher in atopic disease in infancy, but no statistical differences in T-cell subset values between seborrheic and atopic infants were found. T-cell subset values are therefore not a reliable differentiating marker between atopic and seborrheic eczema in infancy. PMID- 2969702 TI - A child with erythematous and hyperkeratotic patches. Erythrokeratodermia variabilis. PMID- 2969703 TI - Expression of CD 21, CD 22, and the mouse erythrocyte receptor on peripheral B lymphocytes in rheumatoid arthritis. AB - The expression of the B cell antigens, CD 21, CD 22, and the mouse erythrocyte receptor (MER), on peripheral mononuclear cells (PMC) in 61 patients with rheumatoid arthritis (RA) and in 25 patients with various other forms of rheumatic disease was studied. Patients with RA showed significantly more peripheral B cells than control patients, whereas there was no difference between patients with RA and controls in resting B cells expressing the MER or resting and activated B cells expressing CD 21. Patients with active RA had significantly less MER positive and more CD 21 positive B cells than patients with inactive disease. The relation between disease activity and expression of MER and CD 21 was independent of drug treatment or production of classical rheumatoid factor. These data may be interpreted as a sign of B cell activation in RA. In addition, patients with seronegative RA receiving gold treatment showed significantly more MER positive cells than patients receiving different drugs, whereas patients receiving non-steroidal anti-inflammatory drugs (NSAIDs) alone had significantly more CD 21 positive cells. This may be the result of different immunomodulating effects of drugs on B cell subsets. PMID- 2969704 TI - Surgery of the descending thoracic aorta: spinal cord protection with the Gott shunt. AB - From July, 1974, to July, 1987, surgical treatment of descending thoracic aortic aneurysms was performed in 173 patients at l'Hopital du Sacre-Coeur de Montreal. The cause of the aneurysms was arteriosclerosis or medial degeneration in 83 patients, trauma in 50, dissection in 34, and a congenital malformation in 6. A single method of external shunting provided distal perfusion in all patients in the series. A 9-mm Gott aneurysm shunt was placed preferentially between the ascending aorta (67%) and the descending aorta (60%). Alternative sites of proximal cannulation (aortic arch, 9%; proximal descending aorta, 22%; left ventricle, 2%) and distal cannulation (abdominal aorta, 3%; left femoral artery, 37%) were chosen based on the location and the extent of the aortic aneurysm. No systemic heparinization was used. In the last 40 patients, a flowmeter adapted for use with the shunt allowed the recording of shunt flow (mean, 2,475 ml/min; range, 1,100 to 4,000 ml/min). Hospital mortality, including patients with ruptured aneurysms, was 15% (26/173). The mean aortic cross-clamp time was 37 minutes (range, 8 to 105 minutes). Of the 173 patients, 168 survived long enough to allow accurate clinical evaluation of the function of the spinal cord: no paraplegia or other spinal cord ischemic injury occurred. To date, our clinical experience has demonstrated the effectiveness of the 9-mm Gott shunt in preserving the functional integrity of the spinal cord during cross-clamping of the thoracic aorta. PMID- 2969705 TI - Revascularization after thrombolytic therapy for acute myocardial infarction: an analysis of 573 patients. AB - From October, 1981, to January, 1987, at our center, 891 patients received streptokinase within 6 hours of acute myocardial infarction. A total of 318 patients were treated medically, while 388 patients (43.5%) underwent coronary artery bypass grafting (CABG) alone and 185 (20.7%) were treated with percutaneous coronary angioplasty (PTCA). Subsequent CABG was performed in 37 of 185 PTCA patients after unsuccessful angioplasty. Group characteristics were similar. However, multiple-vessel coronary artery disease was present in 70.3% of CABG patients compared with 24.1% in the PTCA groups. Procedure mortality was 3.6% for CABG alone, 5.4% for PTCA alone, and 13.5% for the combined angioplasty and operation group (p less than 0.05 compared with CABG). All deaths in the PTCA group with subsequent CABG occurred in those patients taken emergently to CABG (5 of 20 patients). We conclude that with proper patient selection both forms of revascularization are safe and effective. However, emergency coronary bypass surgery in the event of failed angioplasty has a high risk. PMID- 2969706 TI - Effect of experimental cardioplegia methods on normal and hypertrophied rat hearts. AB - The purpose of this study was to evaluate whether the addition of verapamil hydrochloride to oxygenated glucose-rich cardioplegic solution would improve myocardial preservation. The Langendorff preparation of the isolated rat heart was used. Groups of normal (WKY) and hypertrophied (SHR) hearts were treated by five different cardioplegic methods and subjected to 90 or 30 minutes of ischemia at 28 degrees to 29 degrees C and reperfusion at 37 degrees C. The following cardioplegic solutions were used: Group A, cold (16 degrees C) Krebs-Henseleit (KH) glucose free only; Group B, KH with KCL (30 mEq/L) (16 degrees C); Group C, same as B with verapamil (10 microM); Group D, perfusion with oxygenated KH solution containing KCL (30 mEq/L) for 15 minutes prior to ischemia; and Group E, same as D with verapamil (10 microM). Recovery of contraction amplitude, ischemic contracture, coronary perfusate volume, the amount of creatine kinase in the coronary perfusate, heart rate, time of revival, O2 consumption, and ischemic contracture were measured. After 30 minutes of ischemia, we did not find any significant difference among the combinations tested with respect to contraction amplitude recovery. The hearts recovered fully. After 90 minutes of ischemia, we found that the best-protected groups in the normal hearts were Groups D and E. In the hypertrophied hearts, the addition of verapamil to the enhancement solution was harmful. The use of enhancement solution without verapamil prior to ischemia provided the best myocardial protection in the hypertrophied hearts. PMID- 2969707 TI - Technique for repair of left ventricular free wall rupture. AB - A case of postinfarction left ventricular free wall rupture is reported. The technique used to repair the rupture is described, along with a modification of the technique. PMID- 2969708 TI - The left thoracoabdominal incision. AB - The left thoracoabdominal incision is approaching its first century of use. Its history is reviewed. The incision provides wide exposure of the spleen, stomach, left hemidiaphragm, aorta, and esophagus; exposure is not limited to the distal esophagus. Because of the superb exposure it offers, its wide clinical application, and its overall safety, the thoracoabdominal incision should continue to be a viable surgical option in current thoracic surgical practice. PMID- 2969709 TI - [The P wave in systemic arterial hypertension. Comparison with the echocardiogram and apexcardiogram]. AB - The cause of an abnormal electrocardiographic P wave (AEPW) in systemic arterial hypertension (SAH) has not been delucidated. In order to demonstrate if this sign is related to volume overload of the left atrial cavity -previously we found no correlation between pressure overload of the cavity and the presence of an AEPW- 34 patients with SAH were studied; population were divided in two groups: group A (GA, n = 13), formed by 13 cases with a P wave more than 0.10 s duration (D2 standard lead) and group B integrated by 21 cases with a P wave at 0.10 s or less duration. In each cases phonocardiographic and echocardiographic (Mode M) studies were performed and the following data were obtained: "A" index (AI), left atrial dimension (LAD) and, septal (SD) and left posterior wall dimension (LPWD); besides: Sokolow electrocardiographic index (SI) and systolic (SBP) and diastolic (DBP) blood pressure. Results are shown in the following table: (Table: see text). There were not statistical differences between groups (data of SBP and DBP not shown). Correlation between LAD and AI and duration of P wave were not statistical different (data not shown). An AEPW in SAH does not seem to be cause by a volume left atrial overload. Possible it is related to an interatrial conduction defect. PMID- 2969710 TI - [Pre- and postoperative ECG-VCG study of the correction of atrioventricular septal defects]. AB - In order to determine the possible implication of the right bundle branch block (RBBB) in the superior axis deviation of patients (pt) affected with AV septal (Canal) defect, we studied the preoperative and postsurgical ECG of 50 patients who underwent radical correction of the malformation complex. 36 pt had the complete form and 14 a partial form. In 14 of them it was also possible to perform a VCG before and after surgery. The amplitude of S1, R' in a VR and V1 modified significantly (p less than 0.0001), the AQRS showed a clockwise displacement of 19 degrees (p less than 0.002). The terminal forces in the VCG frontal plane changed significantly 62 degrees (p less than 0.0001). Nevertheless the D.I. in a VL and V6 made no difference. Postoperative ECG patterns of pt with partial or complete forms did not differ substantially. Results allowed us to conclude that the major determinant of the AV septal defect ECG pattern is the posterior (inlet) deficiency of the interventricular septum. Surgical correction of the right ventricle overload leads to a clockwise AQRS displacement in the frontal plane. The conduction asynchronism through the left branch showed no modifications. PMID- 2969711 TI - [Normal values in Mexico of the echocardiographic measurements of the left heart]. AB - The M-Mode echocardiographic measurements, according to the international criteria, were done in tracings of 274 normal subjects residents of Mexico City, in order to know the normal values in its population. One hundred and thirty one males (47.8%), and 143 females (52.2%). The age varied from 1 to 73 years, the predominant group was between the third and fifth decades of the life. The age, weight and body surface area (BSA) were correlated with measurements, better correlation was found with BSA and it was the analysis base. According to the BSA, 6 subgroups were formed, the first one of 0.5 m2; increasing 0.4 m2 for each subgroup until the last one of 2.1(2) or more. Were observed proportional amount respect to BSA of D-E mitral valve excursion, aortic root diameter, aortic excursion, aortic opening amplitude, left atrium diameter, end diastolic and end systolic diameters of left ventricle, as well as interventricular septum (IVS) and posterior wall (PW) thickness. The measures were less than the reported other countries. The mean velocity of circumferential shortening (Vcf), fractional shortening (FS) and normalized velocities (NV) showed inverse proportion to BSA. The values of Vcf and NV were similar to the previous report, but the FS and the ejection fraction were higher. The study let us to know the normal echocardiographic values for people of Mexico and the differences with respect to the altitude of this city and the body frame of its population. PMID- 2969712 TI - [Effects of a calcium antagonist (nifedipine) in hypertensive and normotensive subjects]. AB - We investigated the effect of the calcium antagonist nifedipine upon the following parameters: systolic and diastolic blood pressure (SBP and DBP) heart rate (HR), electrocardiogram (ECG) and the relative rate of calcium uptake in platelets. The possible correlation between this rate and blood pressure was one of the main points we tried to establish. The subjects studied were 1) 26 patients with uncomplicated essential hypertension and 2) 13 healthy normotensive subjects. SBP and DBP were measured with the subject both in a recumbent and a sitting position. 10 mg of nifedipine were given orally. In the hypertensive patients SBP and DBP decreased significantly in both positions after receiving the drug, as expected, while HR increased significantly (P less than 0.001), also in both positions. In the normotensive subjects BP decreased too, after taking the drug, but was only significantly modified in some instances i.e. SBP recumbent, DBP recumbent and sitting. HR increased significantly in the sitting position but not in the recumbent position. The ECG post-nifedipine showed a negative depression of the ST segment in four patients from the hypertensive group. The relative rate of calcium uptake in platelets measured before the subjects had taken the drug decreased after it was administered. The difference was significant (P less than 0.05) in the hypertensive group, but not in the normotensive group. Some correlation was found between DBP and the rate of calcium uptake. PMID- 2969714 TI - [Rhabdomyoma in a newborn infant. Echocardiographic diagnosis]. AB - With the 2D echocardiography, is easier the diagnosis of intracardiac tumors nowadays. We present a case of a newborn girl with a rhabdomyoma intra-myocardiac and intracavitary. We discuss the clinical manifestations, surgical indications and the association with tuberous sclerosis. PMID- 2969713 TI - [Angioplasty and percutaneous valvuloplasty in congenital and acquired cardiopathies. Short- and long-term results]. AB - Percutaneous transluminal balloon catheterization was performed in 38 patients, age 2 to 38 years (mean 13) between October, 1985 and May, 1987. Nineteen with pulmonary valve stenosis (PVS), three with small atrial septal defect (ASD) and one with ventricular septal defect (VSD); six with aortic valve stenosis (AVS); eight with Coarctation of the aorta (CoAo), in two with VSD and one with small ductus arteriosus; and four with rheumatic mitral stenosis (RMS). In PVS the peak systolic gradient (PSG) decreased significantly immediately after the procedure (92 +/- 40 vs 20 +/- 19 mmHg P less than 0.01). Nine patients were evaluated three months and seven one year later and no significant change occurred (19 +/- 11 and 20 +/- 19 mm Hg). The right ventricular systolic pressure (RVSP) and the ratio RVSP/left ventricular systolic pressure (LVSP) decreased immediately after the procedure (114 +/- 37 vs 69 +/- 36 mmHg P less than 0.15, and 0.98 +/- 0.29 vs 0.62 +/- 0.36, P less than 0.15). Three months later the RVSP and ratio RVSP/LVSP decreased more (57 +/- 16 mmHg, P less than 0.05, and 0.47 +/- 0.18, P less than 0.05) and one year later (54 +/- 27 mm Hg, P less than 0.05 and 0.46 +/ 0.24, P less than 0.05). In one case we repeated de dilatation one year later for restenosis. In CoAo PSG decreased immediately after dilation (59 +/- 21 vs 26 +/- 18 mmHg, P less than 0.1). Three months later the gradient was 38 +/- 20 mmHg. One patient was sent to surgery for significant residual gradient and one was dilated again 4 months after the first dilatation. One case was evaluated at cardiac catheterization one year later with gradient of 28 mmHg and blood pressure and femoral pulses were normal. In AVS PSG decrease immediately in all (91 +/- 25 vs 37 +/- 22 mmHg, P less than 0.02). Recatheterization in 4 patients three months later showed improvement in two. Two cases were sent to surgery for significant residual gradient. In these two patients we did not performed a dilatation because one had thrombosis in the puncture site and in the other we suspect subvalvular obstruction. The other two patients have shown clinical, echocardiographic and radiographic improvement.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969715 TI - [Color-coded cardiac Doppler]. AB - Color flow mapping with Doppler technique was reviewed. We describe the advantages and technical was reviewed. We describe the advantages and technical limitations of the technique. The usefulness of color flow mapping in acquired and congenital disease was studied. We conclude that new information was added, concerning complex intracardiac flow, in order to the traditional study with classic continuous and pulsed Doppler examination. PMID- 2969716 TI - [Usefulness of M-mode echocardiography, two-dimensional and Doppler, in the study of pericardial diseases]. AB - The objective of this communication is to determine utility of echocardiography M mode, two-dimensional and Doppler, in the diagnosis and follow-up of patients with pericardial disease. This procedure was used initially to the diagnosis of pericardial effusion, actually it has a lot of new applications. It's noninvasive and low cost, and would give us important information about our patients. PMID- 2969717 TI - [Natural history of experimental pulmonary atelectasis in dogs with closed chest]. AB - In order to establish an animal model of pulmonary vasoconstriction we followed the time course of intrapulmonary shunt (Qs/Qt) in a canine model of lobar atelectasis with closed chest. Ten mongrel dogs were studied. Bronchial occlusion of the right lower lobe (RLL) was performed by inflating the balloon of a Foley catheter placed through a rigid bronchoscopy. Analysis of variance was used for statistical analysis. (15 minutes) After occlusion Qs/Qt reached its maximum increasing from 8.2 +/- 3.6 to 29.7 +/- 11.7% (p less than 0.05) and PaO2 decreased from 357 +/- 49 to 100 +/- 43 mm Hg (p less than 0.05). Afterwards, there was a progressive decline of Qs/QT accompanied by an also progressive increase in PaO2. At the end of the experiment (3 hrs post atelectasis) Qs/Qt was 11.2 +/- 4.9 and PaO2 251 +/- 124 mm Hg (p less than 0.05). Pulmonary vascular resistance increased post atelectasis from 439 +/- 168 to 598 +/- 256 d.s.cm-5 (p less than 0.05). Complete atelectasis of the RLL was confirmed postmortem. As the changes in Qs/Qt and PaO2 did not parallel the change in cardiac output we conclude that the mechanism of decrease in Qs/Qt was hypoxic vasoconstriction. PMID- 2969718 TI - [Left ventricular function in physically active and sedentary subjects during physical exercise]. AB - An exercise program based on growing intensity loads (50 watts) was applied to 21 healthy, middle-aged subjects, who were divided into two groups: the physically actives and the sedentaries, according to their physical activity habits. During exercise, which was made on a treadmill, the following variables were measured: oxygen intake (VO2), cardiac frequency (FC), left ventricule ejection time (TEVI), maximal value of the first derivative of ear ensitogram (VMPDDO) and indirect arterial pressure. The active group reached higher values of power and VO2 (absolute and by pulse) and lower values of FC at under maximal works; these data therefore show their best cardiopulmonary capacity. The TEVI in the active group was higher at 150 watts because of the lower FC presented by its assigned subjects. For a maximal exercise condition, both groups had similar ejection times, but the reduction rate of actives' TEVI decreased, which in turn indicates the handling of higher stroke volumes. The VMPDDO relation with the successive increments of work loads was non linear. This index had higher values in the active group, for an exercise strength over 100 watts; this fact allowed the statistical differentiation of both groups. Accordingly, the VMPDDO did indicate the higher left ventricular function of the active subjects during exercise; this findings supports the notion which establishes that resistance training enhances myocardial contraction. In the recovery stage, the sedentaries' VMPDDO increases in a significant way, due to a possible mechanism as to hold stroke volume. In conclusion, the active subjects heart is a more efficient pump than the sedentaries', because of its capacity to handle under maximal exercise higher stroke volumes in the same ejection time, and thus in a higher ejection rate, with equal cardiac frequencies arterial pressures and myocardial oxygen intake. PMID- 2969719 TI - [Critical analysis of 237 cases of elective transluminal coronary angioplasty]. PMID- 2969720 TI - [Re-evaluation of electrocardiographic variables in left ventricular hypertrophy in orovalvular lesions with hemodynamic overload of the left ventricle: chronic aortic and mitral insufficiency and aortic stenosis]. PMID- 2969721 TI - Clinical significance of metabolic disturbances induced by the use of diuretics. PMID- 2969722 TI - The California syndrome. A threat to all. PMID- 2969723 TI - Pharmacological evidence for a role of D2 dopamine receptors in the defensive behavior of the mouse. AB - In this study the role of the DA system in the expression of defensive behavior of the mouse was investigated. C57BL/6 mice subjected to three daily defeat experiences (24 h apart) exhibited an increase of defensive behaviors (upright and sideways postures and escape) as well as a decrease of activity and a decrease of social investigation compared with undefeated mice (controls) when confronted with nonaggressive Swiss mice 24 h after the last aggressive confrontation. The selective D2 DA receptor antagonist (-)-sulpiride administered before confrontation with nonaggressive opponents (fourth day) dramatically decreased defensive behaviors and produced an increase of social investigation. The selective D1 DA receptor antagonist SCH 23390 did not affect either defence or social investigation. In further experiments the behavioral effects of the selective D1 agonist SKF 38393 and of the selective D2 agonist LY171555 on naive C57BL/6 mice interacting with nonaggressive opponents of the same strain were assessed. SKF 38393 in doses up to 30 mg/kg did not produce any significant behavioral changes while LY171555 produced a clear-cut dose-dependent increase of defensive behavior as well as a decrease of social investigation and activity and an increase of immobility. The behavioral profile produced by the D2 agonist did not differ from that produced by defeat experiences. These results indicate that D2 receptors play a major role in the expression of defensive behavior in the mouse. The hypothesis that alteration in D2 receptor functioning may produce hyperdefensiveness possibly due to altered perceptive processes is discussed. PMID- 2969725 TI - Transient-kinetic studies of the adenosine triphosphatase activity of scallop heavy meromyosin. AB - Fluorescence stopped-flow experiments were performed to elucidate the elementary steps of the ATPase mechanism of scallop heavy meromyosin in the presence and in the absence of Ca2+. ATP binding and hydrolysis, as monitored by the change in tryptophan fluorescence, appear to be Ca2+-insensitive, whereas both Pi release and ADP release are markedly suppressed in the absence of Ca2+. Rate constants for Pi release are 0.2 s-1 and 0.002 s-1 and for ADP release are 6 s-1 and 0.01 s 1 in the presence and in the absence of Ca2+ respectively. Ca2+ binding to the specific site of the regulatory domain is rapid and its release occurs at 25 s-1, consistent with the time scale of a twitch of the striated adductor muscle. Nucleotide binding is a multi-step process requiring a minimum of three states. In such a model Ca2+ controls the rate of conformational changes at the active site in both the forward and the reverse direction, leading to a large dependence of the rate of nucleotide release, but a lesser effect on the overall equilibrium position. The kinetic trapping of nucleotides and Pi at the active site, in the absence of Ca2+, appears to be a fundamental step in suppressing the interaction of the myosin head with the thin filaments in relaxed molluscan muscle. PMID- 2969724 TI - The overexpression, purification and complete amino acid sequence of chorismate synthase from Escherichia coli K12 and its comparison with the enzyme from Neurospora crassa. AB - The enzyme chorismate synthase was purified in milligram quantities from an overproducing strain of Escherichia coli. The amino acid sequence was deduced from the nucleotide sequence of the aroC gene and confirmed by determining the N terminal amino acid sequence of the purified enzyme. The complete polypeptide chain consists of 357 amino acid residues and has a calculated subunit Mr of 38,183. Cross-linking and gel-filtration experiments show that the enzyme is tetrameric. An improved purification of chorismate synthase from Neurospora crassa is also described. Cross-linking and gel-filtration experiments on the N. crassa enzyme show that it is also tetrameric with a subunit Mr of 50,000. It is proposed that the subunits of the N. crassa enzyme are larger because they contain a diaphorase domain that is absent from the E. coli enzyme. PMID- 2969726 TI - Kinetic trapping of intermediates of the scallop heavy meromyosin adenosine triphosphatase reaction revealed by formycin nucleotides. AB - The kinetics of interaction of formycin nucleotides with scallop myosin subfragments were investigated by exploiting the fluorescence signal of the ligand. Formycin triphosphate gives a 5-fold enhancement of the emission intensity on binding to heavy meromyosin, and the profile indicates that the kinetics of binding are Ca2+-insensitive. In contrast, the subsequent product release steps show a marked degree of regulation by Ca2+. In the absence of Ca2+ formycin triphosphate turnover by the unregulated and the regulated heavy meromyosin fractions are clearly resolved, the latter showing a fluorescence decay rate of 0.002 s-1, corresponding to the Pi-release step. In the presence of Ca2+ this step is activated 50-fold. Formycin diphosphate release is also regulated by Ca2+, being activated from 0.008 s-1 to 5 s-1. In contrast with protein tryptophan fluorescence [Jackson & Bagshaw (1988) Biochem. J. 251, 515 526], formycin fluorescence is sensitive to conformational changes that occur subsequent to the binding step and demonstrate, directly, an effect of Ca2+ on both forward and reverse rate constants. Apart from a decrease in the apparent second-order association rate constants, formycin derivatives appear to mimic adenosine nucleotides closely in their interaction with scallop heavy meromyosin and provide a spectroscopic handle on steps that are optically silent with respect to protein fluorescence. A novel mechanism is discussed in which regulation of the formycin triphosphate activity by Ca2+ involves kinetic trapping of product complexes. PMID- 2969727 TI - Oligosaccharide mapping of heparan sulphate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane. AB - A new method that we have called 'oligosaccharide mapping' is described for the analysis of radiolabelled heparan sulphate and other glycosaminoglycans. The method involves specific enzymic or chemical scission of polysaccharide chains followed by high-resolution separation of the degradation products by polyacrylamide-gradient-gel electrophoresis. The separated oligosaccharides are immobilized on charged nylon membranes by electrotransfer and detected by fluorography. A complex pattern of discrete bands is observed covering an oligosaccharide size range from degree of polymerization (d.p.) 2 (disaccharide) to approximately d.p. 40. Separation is due principally to differences in Mr, though the method also seems to detect variations in conformation of oligosaccharide isomers. Resolution of oligosaccharides is superior to that obtained with isocratic polyacrylamide-gel-electrophoresis systems or gel chromatography, and reveals structural details that are not accessible by other methods. For example, in this paper we demonstrate a distinctive repeating doublet pattern of iduronate-rich oligosaccharides in heparitinase digests of mouse fibroblast heparan sulphate. This pattern may be a general feature of mammalian heparan sulphates. Oligosaccharide mapping should be a valuable method for the analysis of fine structure and sequence of heparan sulphate and other complex polysaccharides, and for making rapid assessments of the molecular distinctions between heparan sulphates from different sources. PMID- 2969729 TI - Inhibition of lymphocyte proliferation by synthetic peptides homologous to human plasma apolipoproteins B and E. AB - Apolipoproteins B and E of the human plasma lipoproteins are known inhibitors of lymphocyte proliferation. In this report, two synthetic peptide amides, apoB3358 3372 and apoE141-155, showed a dose-dependent inhibition of both the murine mixed lymphocyte culture reaction and the anti-T3 induced proliferation of lymphocytes. Their structures and antiproliferative potencies were similar to that of the heptadecapeptide CKS-17, a consensus peptide of a highly conserved region among HTLV-I, -II and C-type human retroviral proteins. SP-9-2-amide, a peptide homologous to the amino-terminal half of CKS-17, also suppressed lymphocyte activation. In contrast, a peptide homologous to the gp41 protein of HTLV-III that is sequence related to CKS-17 (approximately 35% homology) showed little antiproliferative activity. Neurotensin, a control peptide, showed no activity. The results suggest that a basic tetrapeptide sequence common to CKS-17-amide, SP 9-2, apoB3358-3372 and apoE141-155, but not HTLV-III-amide may account, in part, for the antiproliferative activities of these peptides. PMID- 2969728 TI - Effect of streptozotocin-diabetes on rat liver mitochondrial adenosine triphosphatase turnover. AB - The apparent turnover rates of some mitochondrial enzymes can be modified in diabetes. We studied the effect of streptozotocin-diabetes on the half-life of a protein tightly bound to the inner membrane, ATPase. The half-life (t 1/2), measured by the double-isotope technique, decreased by approx. 20% in diabetes (from approximately equal to 2.56 days in controls to approximately equal to 2.06 days in diabetic rats). These results suggest that diabetes produces an increase in degradation of ATPase by a mechanism which is not yet clear, possibly influenced by alterations induced by diabetes in hepatic lysosomes that are associated with hepatic autophagy. PMID- 2969730 TI - Glipentide and glucose metabolism in isolated rat hepatocytes. AB - Glipentide, a second generation sulfonylurea, raised the cellular concentration of fructose 2,6-bisphosphate in isolated rat hepatocytes. Parallel to accumulating this regulatory metabolite, glipentide inhibited basal gluconeogenesis and increased the rate of L-lactate production, as well as the metabolic flux through the 6-phosphofructo 1-kinase reaction. Tolbutamide elicited similar metabolic effects to those reported for glipentide, although the latter sulfonylurea was about 10 times more potent. The biochemical mechanism by which sulfonylureas promote the accumulation of fructose 2,6-bisphosphate in hepatocytes seems to be related to a significant increase of the hexose 6 phosphate pool (glucose 6-phosphate plus fructose 6-phosphate), together with the activation of 6-phosphofructo 2-kinase and inactivation of fructose 2,6 bisphosphatase, enzyme activities responsible, respectively, for the synthesis and degradation of fructose 2,6-bisphosphate. PMID- 2969731 TI - Interferon gamma is active on human lymphoblastoid Namalva cells without inducing an antiviral state. AB - We demonstrate the presence of high affinity receptors specific for interferon gamma (IFN-gamma) in human lymphoblastoid Namalva cells. The presence of these receptors, whose binding affinity and cross-linking characteristics were not distinguishable from those of the corresponding receptors in sensitive cells, was not consistent with the lack of responsiveness of Namalva cells to IFN-gamma as regards growth inhibition, induction of 2'-5' oligoadenylate synthetase activity and inhibition of virus multiplication. Nevertheless, IFN-gamma enhanced the expression of two genes, HLA class II and c-myc. Although the mechanism of these IFN-gamma-mediated modifications is not understood, these results provide evidence that the IFN-gamma receptors present in Namalva cells are functional. PMID- 2969732 TI - The effect of aldose reductase inhibitor Statil (ICI 128436) on the glucose over utilization in kidney of diabetic rats. AB - The present study examined the effect of the aldose reductase inhibitor Statil (ICI 128436, ICI, Cheshire, U.K.) on the levels of metabolites and activities of enzymes involved in the glycolysis, polyol pathway and pentose phosphate pathway and on the flux of radioactive glucose through these pathways in kidney of streptozotocin diabetic rats. In kidneys of diabetic rats of 30 days duration the level of sorbitol was increased by +82% and fructose concentration was raised by +42%. After treatment with Statil for 9 days (reversal study) a significant fall in kidney sorbitol concentration and kidney fructose concentration was found. Lactate and UDP-glucose concentrations which were both significantly raised in diabetes by +80% and +23% respectively decreased by 20% after Statil treatment, together with a decline in UDP-glucose dehydrogenase activity. Aldose reductase and sorbitol dehydrogenase activities were also significantly lowered by Statil. In the reversal study there was no significant effect of Statil on the flux of glucose via alternative routes in the kidney cortex. In kidneys of diabetic rats of 9 days duration, the level of sorbitol increased by +61% and the concentration of fructose was raised by +30%. The treatment with Statil (25 mg/kg) from the day of induction of diabetes (prevention study) prevented the accumulation of sorbitol, fructose and UDP-glucose. The increase in the incorporation of radioactive glucose through the pentose phosphate pathway seen in diabetes was less marked in the renal cortex of diabetic rats treated with Statil ab initio. PMID- 2969734 TI - [Myocardial hypertrophy, arterial hypertension and sudden cardiac death]. AB - 721 cases of sudden coronary death (SCD) of men aged 30-59 were studied. It was found out that SCD happened in majority of cases in subjects with increased weight of heart: 44.2% of patients had heart weight of 500 g or more. A special histomorphometric study of kidneys (345 cases of SCD) revealed arterial hypertension (AH) in 41.2% of cases, which far exceeded AH incidence in the population of men of equal age. Nevertheless, not in all cases of SCD was myocardial hypertrophy caused by AH. In patients with no AH myocardial hypertrophy was usually caused by postinfarction cardiosclerosis. In certain cases of SCD without cardiosclerosis accompanied by manifest myocardial hypertrophy there were signs of dilatative or hypertrophic cardiomyopathy. The obtained results suggest that AH and increased weight of heart should be considered factors predisposing to SCD. PMID- 2969733 TI - [The initiating role of several respiratory and skin contaminants in food allergies]. AB - The authors present data which suggest that mustard hypersensitivity can result from Isocyanates induced occupational asthma. Isothiocyanates present in mustard spice is probably responsible for this cross-sensitivity. Frequently, food allergies result from skin or respiratory primary contact and sensitization. Some examples are proposed. Distinctive structures and functions of immunological system in cutaneous, respiratory and digestive apparatus, give an explanation for the predominance of skin and respiratory immunological systems to promote hypersensitivity. For the prevention of food allergies, it is very important to avoid excessive skin and respiratory contaminations by allergens in the work and home environments. PMID- 2969735 TI - [Comparative aspects of the structural and functional status of the myocardium in left-ventricular asymmetrical myocardial hypertrophy in patients with hypertension and cardiomyopathy]. AB - 23 patients with hypertrophic cardiomyopathy (HCMP) and 19 patients with stage II of essential hypertension (EH) with left ventricular asymmetric hypertrophy (ASH) were examined. Echocardiography demonstrated significantly more manifest hypertrophy of interventricular septum and higher coefficient of asymmetry in patients with HCMP compared to those with EH while left ventricular myocardium mass was equal in both groups. Phase-volumetric analysis exhibited in all the patients deterioration of myocardial relaxation processes. EKG-monitoring showed a significant increase in frequency of supraventricular rhythm disturbances in patients with HCMP while no reliable differences in total frequency of rhythm and conduction disturbances were registered. Regular adequate antihypertensive therapy resulted in patients with EH in decrease of ASH degree caused by regression of interventricular septum hypertrophy. In natural course of EH the ASH degree also decreased, but in this case because of increased hypertrophy of left ventricular posterior wall. The patients with HCMP were followed for 1-1.5 years. Drug therapy produced no significant changes in left ventricular myocardium mass as well as character of degree of ASH. PMID- 2969736 TI - [Calcium metabolism in arterial hypertension. I. Myocardial hypertrophy and hormone-dependent increase in Ca2+ levels in the thrombocytes of patients with arterial hypertension]. AB - Clonidine-treated patients with essential hypertension demonstrated reliable positive correlations between echocardiographically determined signs of myocardial hypertrophy (thickness of left ventricular posterior wall and interventricular septum, left ventricular myocardial mass) and hormone-dependent increase in thrombocyte calcium concentration induced by ADP and TAF. Antihypertensive therapy was more effective in patients with relatively weak cellular effect of Ca-increasing hormone response having no signs of myocardial hypertrophy. PMID- 2969737 TI - [Comparative pharmacological characteristics of peptides of the atrial natriuretic factor family]. AB - Comparative antihypertensive and natriuretic activities of atrial natriuretic factor (ANF) and its nearest shortened analogues--atriopeptines (AP) were studied in vivo on narcotized rats; and their peripheral vasodilating activities were studied in vitro on isolated rings of rat's thoracic aorta. The results obtained in vivo allow a following series of comparative antihypertensive and vasodilating activity: alpha rANF greater than APII greater than AP7-27. alpha rANF, APIII and APII did not differ significantly in vasodilating activity on isolated vessels; analogue 7-27 was less active. Thus further search of highly active shortened atriopeptin analogues is necessary for developing new drugs on their basis. PMID- 2969738 TI - [Diagnosis of combined myocardial ventricular hypertrophy using precordial mapping. II. Selection of the parameters of the cartogram of diagnostic significance for the differential diagnosis of combined myocardial hypertrophy and its isolated forms]. AB - A new approach to the choice of diagnostic criteria of combined ventricular hypertrophy (CVN) is proposed: after determination of EKG parametres differing from the norm are separately compared to the EKG parametres in isolated left ventricular hypertrophy (LVH) and right ventricular hypertrophy (RVH). The diagnostic parametres were chosen on the basis of computer statistical analysis of EKG-35 data of 136 patients with CVH, 39 with LVH, 57 with RVH and 71 normal subjects. For differentiation from LVH the amplitude/area ratio (S10 + S17 + S24)/(S1 + S8 + S15) and R27/R17 was effectively used, for differentiation from RVH parametres R14/R1 or S25/S27 are recommended. The amplitude parameters provide the sensitivity of the method of 58.8% with the specificity of 94.6%; the use of area ratio of the same waves increases sensitivity up to 72.8% with the specificity of 94.4%. Sensitivity of the method with the use of the Sokolov-Lyon criteria for diagnosis of LVH and RVH in the same groups was 9.6% with the specificity of 94%. PMID- 2969739 TI - Plasma atrial natriuretic peptide in essential hypertension. Comparison with normotensive subjects and effects of changes in dietary sodium intake. AB - Plasma levels of atrial natriuretic peptide (ANP) in 106 patients with essential hypertension with a supine mean blood pressure (mean +/- SEM) of 128.9 +/- 1.6 mmHg and not on treatment were significantly higher than those in 47 normotensive subjects (supine mean blood pressure 93.9 +/- 1.2 mmHg) with mean values of 17.2 +/- 1.1 and 8.6 +/- 0.6 pg/ml, respectively (P less than 0.001). Similar results were found in a subgroup of 35 hypertensive patients identically matched in terms of age, sex, and race with 35 normotensive subjects. Plasma levels of ANP were correlated significantly with age in normotensive subjects and with age and blood pressure in the hypertensive patients. In 12 hypertensive patients studied on a low (10 mmol sodium/day), on their usual sodium intake (around 120 mmol sodium/24 hr) and on a high (350 mmol sodium/day) intake, plasma ANP increased approximately twofold by the fifth day of the high sodium intake, but there was no significant difference between the plasma levels on their usual sodium intake and those on the fifth day of the low sodium intake. Supine mean blood pressure on the patients' usual sodium intake was 119.3 +/- 2.7 mmHg and was reduced to 110.0 +/- 3 mmHg by the fifth day of the low sodium intake (P less than 0.005). However, there was no significant difference between the blood pressure levels on their usual and high sodium intake (118.3 +/- 3.0 mmHg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969740 TI - Effect of doxazosin monotherapy on blood pressure and plasma lipids in patients with essential hypertension. AB - The efficacy and safety of doxazosin (DOX) for the treatment of hypertension was investigated. A multicenter, double-blind, placebo-controlled, parallel design was employed. A 4-week placebo runin period was followed by a 9-week double-blind period during which patients were randomly assigned to placebo or 2, 4, or 8 mg doxazosin. Blood pressures (BP) and heart rates (HR) were measured 24 hours postdose. The mean changes in standing BP (mmHg) were -6.2/-6.9 (2-mg regimen), 5.7/-5.8 (4-mg regimen), -8.5/-7.7 (8-mg regimen) for DOX patients and 0.7/-2.9 for placebo patients. The mean changes in supine BP (mmHg) were -3.2/-4.7 (2-mg regimen), -4.0/-5.1 (4-mg regimen), -4.6/-5.6 (8-mg regimen) for DOX patients and -0.5/-3.3 for placebo patients. There was no evidence of a dose-response relationship for DOX; however, DOX serum levels were linearly related to the dose. Responder rate for the combined DOX patients was 38% (32/84) and for the placebo patients 27% (8/30). HR (24 hours postdose) was not modified by DOX. Patients in the 8-mg regimen had a significantly higher gain in mean body weight (+ 1.3 +/- 0.3 kg; P less than 0.05) compared to the 2-mg regimen, 4-mg regimen, and placebo groups. Plasma norepinephrine was not significantly modified by DOX. DOX had a favorable effect on plasma lipids. DOX lowered LDL cholesterol (P less than 0.05), total cholesterol, and apoprotein B and increased HDL/(LDL + VLDL) ratio (0.05 less than or equal to P less than 0.1) compared to placebo. Dropout rate and treatment-related side effects were equally distributed among the DOX and placebo groups. No patients had the dose of medication reduced because of side effects. Three DOX patients were withdrawn because of postural dizziness. PMID- 2969741 TI - Amelioration of systemic hypertension by converting enzyme inhibition in the renal ablation model. AB - Hypertension associated with a reduction in renal mass has been traditionally thought of as a volume-dependent state. Recent investigations suggest important roles for systemic and glomerular resistance vessels in the pathogenesis of systemic hypertension (SHT) and progression of end-stage renal disease. To examine this relationship, investigations were performed in two groups of rats maintained for 6 weeks following 5/6 renal ablation. Group A received converting enzyme inhibition (CEI) for 6 weeks. Group B received no treatment. Systolic blood pressure and weight of remnant kidney tissue were both increased in group B (P less than 0.01). BUN did not differ in groups A and B; however, renal PGI2 excretion was increased in group A (P less than 0.01). Renal morphology was preserved in group A, with little or no evidence of glomerular sclerosis. CEI prevents SHT and enhances renal PGI2 excretion in this model. The selective increase in PGI2 may mediate systemic and renal effects of this agent. PMID- 2969742 TI - The effects of pressure-induced cardiac hypertrophy on the functional capacity of the coronary circulation. AB - Pressure-induced cardiac hypertrophy has many effects on the functional capacity of the coronary circulation. Many studies have been performed in both animal and humans and the major findings are as follows: 1. Most types of myocardial hypertrophy are associated with the decrement in coronary vasodilator reserve; 2. The magnitude of the decrement in coronary reserve in myocardial hypertrophy is usually much more prominent in patients with myocardial hypertrophy than in animal models; 3. Left ventricular hypertrophy secondary to systemic hypertension is associated with altered autoregulation of myocardial perfusion; 4. The perfusion abnormalities associated with hypertension and left ventricular hypertrophy are affected by various factors such as age of onset, ventricular involvement, and the stimulus for hypertrophy; 5. Left ventricular hypertrophy secondary to renal hypertension markedly augments the adverse effects of coronary occlusion. In this setting, coronary occlusion is associated with a three-fold increase in the incidence of lethal ventricular arrhythmias and a 35% increase in infarct size. Thus, pressure-induced hypertrophy profoundly alters the coronary circulation. PMID- 2969743 TI - Blood pressure during long-term converting-enzyme inhibition predicts the curability of renovascular hypertension by angioplasty. AB - Percutaneous transluminal renal angioplasty (PTRA) was performed in 28 hypertensive patients with 50% or more unilateral renal artery stenosis. Prospectively, the study compared the extent to which systolic (SBP) and diastolic (DBP) pressure during long-term converting-enzyme inhibition (CEI) and the ipsi- to contralateral renal vein renin ratio (RVR) predicted success of PTRA, defined as SBP less than 160 mmHg and DBP less than 95 mmHg. Both SBP and DBP after PTRA were positively correlated with pressure during CEI (P less than 0.001) and negatively with RVR (P less than 0.03). In multiple regression these associations were independent and remained statistically significant. In discriminant analysis, a SBP during CEI less than 160 mmHg, a DBP during CEI less than 95 mmHg, and a RVR greater than or equal to 1.5 identified with equal accuracy the success of PTRA. Furthermore, in patients with a SBP during CEI less than 160 mmHg, the demonstration of a RVR greater than or equal to 1.5 increased (P less than 0.05) the prediction of a positive outcome from 50 to 67%. In contrast, in 92% of the patients with a SBP during CEI greater than 160 mmHg, PTRA was not successful. Thus, blood pressure measurements during long-term CEI predict the curability by PTRA of renovascular hypertension and can be employed either alone or in association with the RVR. PMID- 2969744 TI - Associative encoding and retrieval in Alzheimer's and Huntington's disease. AB - The associative encoding and retrieval abilities of patients with dementia of the Alzheimer type (DAT) and patients with Huntington's disease (HD) were investigated using an encoding specificity paradigm. When compared to age- and education-matched intact control subjects, both patient groups were severely (and equally) impaired on overall memory for word lists. However, the HD and DAT patients showed differential improvement in recall performance with the introduction of associated cues during stimulus presentation and recall. Although the HD patients, like intact subjects, were able to benefit from semantic retrieval cues (strong and weak) which were present during input, the performance of the patients with DAT improved only with the introduction of strong cues at output, regardless of whether the cues were present or absent during initial presentation. These findings suggest that patients with DAT failed to encode the semantic relationship between the to-be-recalled and cue words and simply generated free associations to the cue words during retrieval. Similarities between the performances of the patients with DAT and alcoholic Korsakoff patients are noted and discussed with regard to the neuropathological basis of the disorders. PMID- 2969745 TI - Attendance patterns for dental treatment of inoculation risk patients. PMID- 2969746 TI - Linear fixed drug eruption. PMID- 2969747 TI - Pre-operative treatment of an abdominal wall endometrioma with nafarelin acetate. Case report. PMID- 2969748 TI - Trypsin digestion of junctional sarcoplasmic reticulum vesicles. AB - A putative constituent of the junctional processes, connecting the terminal cisternae of sarcoplasmic reticulum and the transverse tubules of skeletal muscle fibers, is a greater than or equal to 350,000-dalton (Da) protein that displays ryanodine binding and Ca2+ channel properties. Ryanodine modulation of Ca2+ fluxes suggests that the ryanodine receptor and calcium channel are integral parts of one functional unit corresponding to the greater than or equal to 350,000-Da protein [Inui, M., Saito, E., & Fleischer, S. (1987) J. Biol. Chem. 262, 1740-1747; Campbell, K. P., Knudson, C. M., Imagawa, T., Leung, A. L., Sutko, J. L., Kahl, S. D., Raab, C. R., & Madson, L. (1987) J. Biol. Chem. 262, 6460-6463]. We subjected vesicular fragments of junctional-cisternal membrane to stepwise trypsin digestion. The greater than or equal to 350,000-Da protein is selectively cleaved in the early stage of digestion, with consequent disappearance of the corresponding band in electrophoretic gels. The Ca2+-ATPase is cleaved at a later stage, while calsequestrin is not digested under the same experimental conditions. While the Ca2+-ATPase yields two complementary fragments that are relatively resistant to further digestion, the greater than or equal to 350,000-Da protein yields fragments that are rapidly broken down to small peptides. Under conditions producing extensive digestion of the greater than or equal to 350,000-Da protein, the junctional processes are still visualized by electron microscopy, with no discernible alterations of their ultrastructure. The functional properties of the Ca2+ release channel are also maintained following trypsin digestion, including blockage by Mg2+ and ruthenium red and activation by Ca2+ and nucleotides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969749 TI - Bovine plasminogen activator inhibitor 1: specificity determinations and comparison of the active, latent, and guanidine-activated forms. AB - The plasminogen activator inhibitor 1 (PAI-1) synthesized and released by cultured bovine aortic endothelial cells is present in conditioned medium in a latent form that can be activated by guanidine hydrochloride [Hekman, C. M., & Loskutoff, D. J. (1985) J. Biol. Chem. 260, 11581-11587]. The purified, guanidine activated PAI-1 was shown to inhibit both plasmin and trypsin in a dose- and time dependent manner. Second-order rate constants for these interactions were calculated to be 6.6 X 10(5) and 7.0 X 10(6) M-1 s-1 for plasmin and trypsin, respectively. Experiments were conducted to compare the inherently active and the guanidine-activated forms of PAI-1. The two active forms had similar kinetic parameters for interaction with urokinase (Kd, 0.3 pM; kassoc, 1.5 X 10(8) M-1 s 1) and were both inactivated upon treatment with acid or base and by incubation at 37 degrees C. The latent form was relatively stable when incubated under similar conditions. The decrease in PAI-1 activity upon incubation at 37 degrees C was partially restored by a second treatment with guanidine hydrochloride. However, the degree of recovery decreased as a function of incubation time at 37 degrees C. These data suggest that active and guanidine-activated PAI-1 represent a single form of PAI-1. Incubation of this form at 37 degrees C yields two distinct populations of inactive PAI-1, one capable of reactivation and another that appears to be irreversibly inactivated. PMID- 2969750 TI - Two-step internalization of Ca2+ from a single E approximately P.Ca2 species by the Ca2+-ATPase. AB - Phosphorylation by ATP of E.*Ca2 (sarcoplasmic reticulum vesicles (SRV) with bound 45Ca2+) during 5-10 ms leads to the occlusion of 2 *Ca2+/EPtot [quench by ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) alone] in both "empty" (10 microM free Ca2+in) or "loaded" SRV (20-40 mM free Ca2+in). The rate of Ca2+ "internalization" from the occluded E approximately P.*Ca2 was measured by using an ADP + EGTA quench; a *Ca2+ ion that is not removed by this quench is defined as internalized. In the presence of 20-40 mM unlabeled Ca2+ inside SRV, 1 *Ca2+/EPtot is internalized from 45Ca-labeled E approximately P.*Ca2 with a first-order rate constant of kl = 34 s-1. Empty SRV take up 2 *Ca2+/EPtot with the same initial rate, but the overall rate constant is kobsd = 17 s-1. The apparent rate constant (kb = 17 s-1) for internalization of the second *Ca2+ is inhibited by [Ca]in, with K0.5 approximately 1.3 mM and a Hill coefficient of n = 1.1. These data show that the two Ca2+ ions are internalized sequentially, presumably from separate sequential sites in the channel. [32P]EP.Ca2 obtained by rapid mixing of E.Ca2 with [gamma-32P]ATP and EGTA disappears in a biphasic time course with a lag corresponding to approximately 34 s-1, followed by EP* decay with a rate constant of approximately 17 s-1. This shows that both Ca2+ ions must be internalized before the enzyme changes its specificity for catalysis of phosphoryl transfer to water instead of to ADP. Increasing the concentration of ATP from 0.25 to 3 mM accelerates the rate of 45Ca2+ internalization from 34 to 69 s-1 for the first Ca2+ and from 17 to 34 s-1 for the second Ca2+. High [ATP] also accelerates both phases of [32P]EP.Ca2 disappearance by the same factor. The data are consistent with a single form of ADP-sensitive E approximately P.Ca2 that sequentially internalizes two ions. The intravesicular volume was estimated to be 2.0 microL/mg, so that one turnover of the enzyme gives 4 mM internal [Ca2+]. PMID- 2969751 TI - Characteristics of a Ca2+-ATPase activity measured in islet homogenates. AB - Ca2+-ATPase activity was measured in rat islet homogenates, in a medium of low ionic strength containing a low concentration of Ca2+ and Mg2+ and devoid of K+. The enzyme activity was highly sensitive to inhibition by compound 48/80 (a calmodulin inhibitor), stimulated by 120 nM calmodulin and slightly affected by 10 mM NaN3. The addition of Mg2+ to the assay medium promotes the disappearance of apparent Ca2+-ATPase activity. Ouabain (0.1 mM) did not modify this ATPase activity. The enzyme showed two kinetic components for Ca2+ as well as for ATP: one with high apparent affinity and low maximum velocity and the other with low apparent affinity and high maximum velocity. Incubation of islet homogenates in this assay medium with [gamma-32P]ATP in the presence of proteolytic inhibitors, results in the appearance of a single labelled band of 130 kDa, identified by gel electrophoresis. The incorporation of 32P into this band was similar in the presence of either 2.8 or 50 microM Ca2+ and susceptible to hydroxylamine attack. The results indicate that, under the conditions described above, the Ca2+-ATPase activity evidenced in the islet homogenates had characteristics resembling those of the enzyme which catalyzes the outward Ca2+ transport. On the other hand, the method could provide a useful tool to test the effect of different agents which affect insulin secretion upon the islet plasma membrane Ca2+-ATPase activity. PMID- 2969752 TI - Effect of divalent cation chelation on dihydropyridine binding in isolated cardiac sarcolemma vesicles. AB - The effect of divalent cation chelation on specific nitrendipine and ouabain binding has been determined in a highly enriched sarcolemma preparation isolated from canine ventricle. Maximal high-affinity nitrendipine binding measured in the absence of added calcium or magnesium was 997 +/- 103 fmol/mg protein. Nitrendipine binding in the presence of EDTA significantly decreased to 419 +/- 42 fmol/mg protein (P less than 0.001) which equates to 42.0% of control. The simultaneous presence of EDTA and A23187 in the binding buffer resulted in a decrease in nitrendipine binding to below detectable levels. These results suggest that divalent cations trapped within vesicles can support high affinity nitrendipine binding. Evaluation of dihydropyridine binding at various pH values suggested that the loss of binding below pH 7.0 and above pH 8.0 may result indirectly from a change in divalent cation binding rather than a direct effect on dihydropyridine binding per se. The maximal binding of ouabain determined in the presence of magnesium and inorganic phosphate averaged 340 +/- 7.4 pmol/mg protein. Pre-treatment of the preparation with sodium dodecyl sulfate (SDS) in order to express binding in sealed inside-out (IO) vesicles, increased ouabain binding to 471 +/- 27 pmol/mg protein. Thus, these preparations averaged 27.8% sealed IO vesicles. Addition of EDTA in the absence of magnesium in the binding buffer reduced ouabain binding to 204 +/- 7.7 and 11.7 +/- 3.5 pmol/mg protein in control and SDS-treated preparations, respectively. These findings suggest that this sarcolemma preparation consists of 43.6% sealed right-side-out (RO) vesicles which contain sufficient endogenous divalent cation trapped in the intravesicular space, to support ouabain binding. The correspondence between the percentage of ouabain binding that remains in the presence of EDTA and the percentage of nitrendipine binding observed under the same conditions is consistent with the hypothesis that divalent cations support nitrendipine binding by interaction with a site or sites accessible only from the cytoplasmic membrane surface and that nitrendipine and ouabain binding sites occur in the same vesicles (i.e., the nitrendipine binding site is of sarcolemma origin). PMID- 2969753 TI - Processing reactions in the later stages of hormone activation. AB - Three tiers of processing have been investigated in the reactions that transform prohormones into their mature end products. Evidence is presented that the proteolytic reactions that convert lipotropin into shortened forms of beta endorphin take place in individually distinct stages. After these cleavages have occurred, the removal of basic residues by carboxypeptidase H and amidation of the products are effected by independent reactions which do not synergise. Experiments are also described which show that the amidating enzyme can accept certain imino acids as substrates and utilises a mechanism that involves hydroxylation; it is implicit that peptide amidation proceeds by a similar mechanism. These results point to a general concept that pro-hormone processing involves consecutive reactions which take place in a predetermined order. PMID- 2969754 TI - Structural and immunological homology of human and porcine pituitary and plasma IRCM-serine protease 1 to plasma kallikrein: marked selectivity for pairs of basic residues suggests a widespread role in pro-hormone and pro-enzyme processing. AB - IRCM-serine protease 1 (SP1), originally isolated from porcine pituitaries and exhibiting preference for cleavage at pairs of basic residues has now been isolated in sufficient quantities to be structurally characterized from both porcine and human pituitaries and plasmas. Whereas the porcine protease shows a high degree of amino acid sequence homology to human plasma pre-kallikrein, the human homologue exhibits an identity of sequence in the first 25 residues of each chain (regulatory and catalytic chains). In addition, human plasma and pituitary IRCM-SP1 and human plasma pre-kallikrein show virtually identical immunological and molecular properties. These data strongly suggest that IRCM-SP1 and plasma pre-kallikrein originate from the same gene product. Purified extracts from perfused rat pituitaries show that 32% of the IRCM-SP1 activity found in normal rat pituitaries, still remain. These data together with the demonstrated association of IRCM-SP1 with particulate fractions of the pituitary suggest that IRCM-SP1 represents a tissue form of plasma pre-kallikrein. The characterization of the digestion products obtained upon reaction of IRCM-SP1 with pro-insulin, ACTH1-39, pro-dynorphin and pro-enkephalin-derived peptides, somatostatin-28, and a pro-renin-like peptide confirmed the high degree of cleavage selectivity of this enzyme for pairs of basic residues. PMID- 2969756 TI - [Ecological characteristics of the structural organization of respiratory segments of the lung]. AB - The structure of the bronchial tree in the lung lobules has been studied and the number of lobules and acini in different lobes and in the entire lung has been determined in polar foxes and dogs. Branching pattern and the number of generations of interlobular and intralobular bronchioli have been defined. It has been found that the number of lung lobules in polar foxes was twice as high as in dogs, the lobule volume being smaller almost by half. A smaller number of acini in polar fox lobules enables the central regulation mechanisms to intensify the control of local ventilation. It broadens the possibilities for physiological responses to the influence of climatic factors of the Extreme North. PMID- 2969755 TI - Regional and subcellular localization in human brain of [3H]paroxetine binding, a marker of serotonin uptake sites. AB - The characteristics of the binding of [3H]paroxetine, a selective serotonin (5 HT) uptake blocker, were investigated in human brain. The Kd value was 0.23 +/- 0.07 nM, and the Bmax value was 190 +/- 39 fmol/mg protein in the putamen. The capacity of various antidepressive drugs to inhibit [3H]paroxetine-specific binding in human brain was well correlated with their capacity to inhibit [3H]5 HT uptake in rat brain. The highest concentrations of [3H]paroxetine-specific binding sites were found in the substantia nigra, hypothalamus, and hippocampus. Lower values were obtained in the basal ganglia and the thalamus. The specific binding was very low in cerebral and cerebellar cortices. The regional distribution of [3H]paroxetine binding sites differs from that of [3H]ketanserin binding to S2 serotonin receptors. The subcellular distribution of the [3H]paroxetine-specific binding sites obtained by differential centrifugation revealed a synaptosomal enrichment in the frontal cortex and striatum, whereas an enrichment in the microsomal fraction was found in striatum. The results show that [3H]paroxetine is a ligand of choice to label the 5-HT uptake molecular complex in human brain. PMID- 2969757 TI - [Selective anticonvulsive action of N-substituted imidazole-4,5-dicarboxylic acids against quinolinic acid]. AB - Selective antagonists of quinolinic acid (2,3-pyridine dicarboxylic acid, QUIN)- an endogenous convulsant tryptophan metabolite, administered intracerebroventricular to mice, were identified during comparison with the following intracerebroventricular convulsants: l-kynurenine, aspartic, glutamic, N-methyl-DL-aspartic and kainic acids. It is suggested that the antagonism arises due to a common fragment of the structure which consists of two carboxylic groups at two nearest carbon atoms of the ring and of one nitrogen atom in the alpha position. The selective action of the compounds found against QUIN supports the suggestion that QUIN produces seizures via N-methyl-D-aspartate binding sites. PMID- 2969759 TI - Criteria for ankylosing spondylitis: facts and fallacies. AB - Weaknesses in existing clinical and radiological criteria for diagnosing ankylosing spondylitis are highlighted and suggestions for improvement made. It is suggested that a major stride forward would be to incorporate a system of probability weighting for individual criteria. This would not only allow diagnosis to be expressed in terms of statistical confidence but would also allow diagnosis to be codified. This latter could facilitate the interpretation of data. PMID- 2969758 TI - The human T-cell V gamma gene locus: cloning of new segments and study of V gamma rearrangements in neoplastic T and B cells. AB - The authors have analyzed the involvement of V gamma and J gamma segments in TRG gamma rearrangement from a series of 40 acute lymphoblastic leukemia (ALL), including 25 T- and 15 B-lineage cases, in which TRG gamma are rearranged. Sixty five rearranged alleles were studied. The authors first describe the cloning and sequencing of two variable segments, V gamma 11 and psi V gamma 12, which rearrange in T- and B-neoplastic cells. To date three subgroups of translatable V gamma segments have been described. The authors show that V gamma 11 is the unique member of a new fourth V gamma subgroup that also rearranges in normal polyclonal T cells and that psi V gamma 12 is located at 5-kilobase (kb) downstream to V gamma 11. As shown by DNA sequence analysis, V gamma 11 shares a 60% homology with V gamma 10 (third subgroup) and a 50% homology with V gamma 9 (second subgroup) but no appreciable homology with the V gamma segments from the first family. In contrast to psi V gamma 12, V gamma 11 is translatable. In this paper the authors have also attempted to determine which V gamma segments were rearranged in the ALL cases by hybridization with a J gamma probe and genomic probes specific of the four subgroups. In the 54 instances in which the rearrangement was consistent with J gamma 1 or J gamma 2 involvement, the authors have identified the corresponding V gamma segments and have not found any other rearrangements suggestive of the existence of further V regions. The V gamma segments, belonging to the first subgroup, were the most frequently used (41 alleles). V gamma 9, V gamma 10, V gamma 11, and psi V gamma 12 were found rearranged in cases 3, 4, 5, and 1, respectively. No cases using the pseudo psi V gamma 1, psi V gamma 5, and psi V gamma 6 segments were found. Pseudo V gamma segments were not found rearranged in T cells, while V gamma 2 and V gamma 4, segments are frequently used. In contrast to the V gamma I gene rearrangement, the involvement of the V gamma II, V gamma III, and V gamma IV subgroups was most frequently observed in T-ALL with stage II differentiation (CD7+, CD4+, and/or CD8+, CD3-), than in those with stage I (CD7+, CD4-, CD8-, CD3-), than in those with stage I (CD7+, CD4-, CD8-, CD3-) and stage III (CD7+, CD4+/-CD8+/ CD3+).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969760 TI - Care of physically handicapped young adults. PMID- 2969761 TI - Which patients should undergo laparoscopy? PMID- 2969762 TI - Procedural learning and neostriatal dysfunction in man. AB - Patients with early stage Parkinson's disease are shown to be selectively impaired in a cognitive task of procedural learning while remaining intact in recall and recognition tests of declarative memory. In contrast, amnestic patients showed the opposite set of deficits, thus demonstrating a double dissociation. Patients with early Huntington's disease were either comparable to the parkinsonian patients or to amnestics. In the advanced Huntington's group, both procedural learning and declarative memory were impaired. It is argued that cognitive procedural learning depends on the establishment of heuristic strategies through the action of a circuit which involves the neostriatum and the prefrontal cortex. PMID- 2969763 TI - Vasopressin stimulates inositol phospholipid metabolism in rat medulla oblongata in vivo. AB - We have investigated whether microinjections of arginine-vasopressin (AVP) into the nucleus tractus solitarii (NTS) of rats results in the stimulation of inositol phospholipid metabolism, a biochemical event related to the activation of V1-type receptors. We found that AVP, but not the related peptides oxytocin or deamino-D-AVP, increases the incorporation of [3H]myoinositol into lipids and the accumulation of labelled inositol phosphates in the dorsomedial medulla oblongata of lithium-treated rats. These effects were abolished by local pretreatment with D-(CH2)5-Tyr(Me)-AVP, a specific antagonist of V1-receptors. It is concluded that the effects of AVP in the NTS of rats are likely to be due to an action on specific receptors (V1-like) and subsequent activation of polyphosphoinositide turnover. PMID- 2969764 TI - Quinolinic acid lesions of rat striatum abolish D1- and D2-dopamine receptor mediated catalepsy. AB - The selective D1-dopamine receptor antagonist SCH23390 and the more D2-selective antagonist haloperidol produced marked catalepsy in rats. The novel excitotoxin quinolinic acid (QA) selectively destroys striatal neurons when injected directly into the striatum. Bilateral QA lesions of the rat striatum (150 nmol and 225 nmol per side) abolished the cataleptic response to both SCH23390 and haloperidol. These data indicate that the D1- and/or D2-dopamine receptors which mediate the cataleptic response are restricted to QA-sensitive neurons in the rat striatum. PMID- 2969765 TI - Identification of D1-dopamine receptor in chicken embryo retina with [125I]SCH 23982. AB - The D1-dopamine receptor in chicken embryo retina was identified with the D1 dopamine receptor specific ligand, [125I]SCH 23982. Binding of [125I]SCH 23982 to both pre-hatched and post-hatched chicken retina was rapid, saturable and of high affinity. The dissociation constant and maximal binding capacity were 795 +/- 25 pM (mean +/- S.E.M., n = 3) and 32.2 +/- 3.8 fmol/mg protein (mean +/- S.E.M., n = 3), respectively for 13-day-old chicken embryo retina, and 785 +/- 58 pM (mean +/- S.E.M., n = 3) and 96.9 +/- 4.1 fmol/mg protein (mean +/- S.E.M., n = 3), respectively for 1-day-old post-hatched chicken retina. The binding properties of the D1-dopamine receptor in chicken retina were similar to those in rat striatum. The maximal binding capacity of the D1-dopamine receptor for [125I]SCH 23982 was increased concomitant with embryonic development, but without any changes in either affinity or pharmacological properties. Dopamine-stimulated adenylate cyclase activity in the retinal homogenates increased concomitant with embryonic development, diminished in the presence of 1 microM SCH 23390 (a D1-dopaminergic antagonist) but remained unaffected by 1 microM YM-09151-2 (a D2-dopaminergic antagonist). PMID- 2969766 TI - Blockade of in vivo binding of 125I-labeled 3-iodobenzamide (IBZM) to dopamine receptors by D2 antagonist and agonist. AB - The in vivo binding of [125I]3-iodobenzamide (IBZM), a substituted benzamide, to DA receptor binding sites in the caudate nucleus, nucleus accumbens, and olfactory tubercle was investigated by using ex vivo autoradiography. The in vivo binding of IBZM seems to be selective to D2 dopamine receptors, since the binding was blocked by pretreatment of animals with D2 agonist LY-171555 or antagonist YM 09151-2. Furthermore, in vitro binding assays in striatal membranes confirmed that IBZM binding was highly selective to D2 sites. Thus, IBZM, when labeled with 123I (T1/2: 13h; 159 kev), could be a potential ligand for imaging D2 dopamine receptors by single photon emission computerized tomography procedures. PMID- 2969768 TI - Behavioral and biochemical expression of D1-receptor supersensitivity following SCH 23390 repeated administrations. AB - In this study concomitant changes of behavioral and biochemical responses in rats repeatedly treated with SCH 23390 were evaluated. Apomorphine-induced stereotyped behavior was increased in D1-supersensitive rats, in contrast no change in the behavioral response to the pure D1-agonist SKF 38393 was detected. Parallel biochemical studies indicated that the enhancement in striatal dopamine (DA) metabolite concentration due to the administration of spiroperidol plus SKF 38393 was not potentiated in SCH 23390 repeatedly treated rats. The results show that the expression of D1-supersensitivity could depend on the stimulation of D2 receptors. PMID- 2969767 TI - Anesthetic effects on 5-hydroxytryptamine uptake by rat brain synaptosomes. AB - As a neurotransmitter involved in modulating central nervous system nociception and awareness, 5-hydroxytryptamine (5-HT) may play an important role in the clinical sequelae of certain anesthetic compounds. Anesthetic agents are known to affect peripheral, non-neuronal 5-HT uptake but little is known about their effects on 5-HT metabolism in the central nervous system. The effects of several anesthetic compounds on 5-HT uptake were examined in synaptosomes isolated from rat brain cortex. Inhibition of this uptake process was observed by exposure to clinically relevant concentrations of the volatile anesthetics halothane, isoflurane, and enflurane. The non-volatile agent, ketamine also inhibited uptake while the narcotic fentanyl had an effect only at the highest concentrations tested. Non-volatile agents which had neither a consistent nor significant effect on synaptosomal 5-HT uptake included pentobarbital, sufentanil, and etomidate. These alterations of 5-HT metabolism could represent a mechanism that contributes to anesthetic action. PMID- 2969769 TI - Treatment of seriously ill and handicapped newborns. AB - Medical technology makes it increasingly possible to prolong life, sometimes indefinitely in the face of overwhelming disability or suffering; thus, choices must be made whether to withhold or withdraw life-sustaining treatment. Reasonable people disagree about who should make such decisions when the patient is an infant, and what principles or rules should guide them. The article briefly reviews the recent history of this controversy analyzes the major ethical substantive principles that have been proposed to guide conduct, and presents the arguments for a procedural approach that is now attracting interest. PMID- 2969770 TI - Ethanol and carbon-source starvation enhance the accumulation of HSP80 in Neurospora crassa. AB - In Neurospora crassa, heat shock results in the induction of 9 to 11 heat shock proteins (HSP), of which HSP80 is the most abundant and the first to be synthesized. The induction of HSP80 was investigated during normal growth (2% sucrose) and under sucrose starvation. Transfer of mycelium to a medium supplemented with ethanol stimulated the synthesis of HSP80, even at the normal growth temperature of 28 degrees C. It was also synthesized under carbon starvation conditions, where the medium was supplemented with 0.02% sucrose, 0.3% acetate, 0.2% lactate, or ethanol. A 30-35 kilodalton polypeptide was induced by heat shock in carbon-sufficient media, but in 0.02% sucrose and 0.3% acetate containing media it was synthesized at normal temperatures. While the overall heat shock response remained unaltered in these cultures, the abundance of HSP90 and HSP70, relative to HSP80, was greater. HSP80 appears to be controlled by carbon-catabolite repression as well as heat shock. Another high molecular mass protein (tentatively designated alc'80') was observed to be induced by heat shock, provided carbon starvation conditions prevailed concurrently. PMID- 2969771 TI - Individualization of patient care using Orem's theory. PMID- 2969772 TI - Plasma cortisol, prolactin, growth hormone, and immunoreactive beta-endorphin response to fenfluramine challenge in depressed patients. AB - The effect of a single dose (60 mg p.o.) of the serotonin agonistic agent fenfluramine (FNF) on plasma cortisol, prolactin (PRL), growth hormone (GH), and immunoreactive beta-endorphin (ir-beta-EP) levels was assessed in eight major depressed patients and eight controls. The hormones were monitored at basal level (0') and hourly during 5 h following FNF administration. The pharmacological challenge caused an elevation of 80% in PRL secretion in the healthy controls and only 42% in the depressed patients. However, the actual prolactin response (delta max) failed to discriminate depressed patients from controls. A blunted response followed by a decrease (33%) in serum cortisol levels was observed in depressed patients 5 h after drug administration while an increase of 94% was obtained in controls after 3 h. FNF provocation did not affect GH and ir-beta-EP plasma levels. The blunted cortisol responsiveness to FNF administration in depressed patients may reflect functional hypoactivity of central serotonergic system at least during the acute phase of major depression. It is not clear why the cortisol hyporesponsivity in depressed patients is not accompanied by a similar reduced PRL response to FNF challenge. PMID- 2969773 TI - Antigen processing and CD4+ T cell depletion in AIDS. PMID- 2969774 TI - Analysis of host-virus interactions in AIDS with anti-gp120 T cell clones: effect of HIV sequence variation and a mechanism for CD4+ cell depletion. AB - The primary human T cell response to HIV was analyzed by isolating from seronegative donors T cell clones specific for HIV gp120. T cell epitopes restricted by different MHC elements were identified within gp120, and synthetic peptides were used to address the fundamental problem of how HIV sequence variability affects T cell recognition. Even one conservative substitution can drastically reduce recognition; thus the interaction of gp120 epitopes with T cell receptors and MHC is precise and poorly crossreactive. Importantly, a subset of CD4+ gp120-specific clones manifest cytolytic activity and lyse uninfected autologous CD4+Ia+ T cells in the presence of gp120 in a process that is strictly dependent upon CD4-mediated uptake of gp120 by T cells. Assuming gp120 is shed from HIV-infected cells in vivo, this novel CD4-dependent autocytolytic mechanism may contribute to the profound depletion of CD4+ cells in AIDS. PMID- 2969775 TI - Decline in the production of interleukin-3 with age in mice. AB - Previously, we and others have found that the ability to produce interleukin-1 (IL-1) and interleukin-2 (IL-2) declines with age in mice. The purpose of this study was to determine the influence of age on the capacity of mice to produce interleukin-3 (IL-3). Splenic cells (5 X 10(6)/ml) from young (3-4 months) and old (24-32 months) C57BL/6 mice were first assessed for their IL-3-producing capacities in response to varying doses of concanavalin A (Con A; 2-20 micrograms/ml) in a time-dependent manner. The results showed that the production of IL-3 by both young and old C57BL/6 mice was maximal on Days 3 and 4 in response to 20 micrograms/ml of Con A, and that of IL-2 was minimal (activity was less than 0.1 unit) on Day 4. Consequently, Day 4, was selected to assess the effect of age on IL-3 production by splenic cells. The results showed a twofold reduction in IL-3 production with age (P less than 0.05). Young-old splenic cell mixture experiments at ratios of 1:0, 3:1, 1:1, 1:3, and 0:1 indicated that the decrease in IL-3 production with age was not due to an increase in suppressor cell activity. Experiments based on mixtures of nylon wool-enriched splenic T cell and adherent cells and on anti-MAC-1 plus complement-treated spleen cells indicated that (a) adherent cells are not required for T-cell production of IL-3, unlike IL-2 production, and (b) the decrease in IL-3 production with age is due solely to alteration in IL-3-producing T cells. Finally, a strong correlation was demonstrated between the production of IL-2 and IL-3 by spleen cells of individual young and old mice (r = 0.92, P less than 0.01). That production of both IL-2 and IL-3 is affected in a similar manner by age would suggest that a single class of helper T cells may be responsible for production of both lymphokines. PMID- 2969776 TI - Monocyte-independent interleukin-2 production and proliferation of human T cells in response to murine hybridomas expressing the OKT3 monoclonal antibody: interleukin-1 is not required for T-cell proliferation. AB - We report a new, monocyte-independent system for the induction of activation and proliferation of human T cells in response to murine hybridomas expressing the OKT3 monoclonal antibody (OKT3 hybridomas). Incubation of nylon-wool-nonadherent (NA) lymphocytes or purified T cells with OKT3 hybridomas resulted in interleukin 2 (IL-2) production, expression of IL-2 receptor, modulation of the CD3 antigen, and proliferation. In contrast, murine hybridomas (OKT4, OKT8, anti-HLA-DR, and others) expressing monoclonal antibodies (mAb) other than OKT3 did not induce T cell activation and proliferation. T cells did not respond to OKT3 mAb alone. OKT3 hybridomas alone did not produce interleukin-1 (IL-1) or other soluble factors that might be involved in the induction of IL-2 production by T cells, and they did not contain membrane-bound IL-1. In addition, IL-1 activity was not detected in cultures of NA-lymphocytes and OKT3 hybridomas, clearly demonstrating that IL-1 was not required, at least in this system, for T-cell activation and proliferation. Direct cell-cell contact between T cells and OKT3 hybridomas was required for IL-2 production. Thirty to fifty percent of T cells formed conjugates with the OKT3 hybridomas but not with the OKT4 or OKT8 hybridomas. Both conjugate formation and IL-2 production were significantly inhibited by the OKT3 mAb and by the anti-LFA-1 mAb. The cells responsible for IL-2 production were found to be of the T3+ T4+ T8- Leu 7- Leu 11- phenotype. IL-2 activity produced by NA-lymphocytes in response to OKT3 hybridomas became detectable as early as 1 hr and reached a maximum by 8 hr, preceding IL-2 receptor expression, modulation of the CD3 antigen, and [3H]thymidine incorporation of T cells. T cells produced higher concentrations of IL-2 in response to OKT3 hybridomas than in response to equal numbers of monocytes and OKT3 mAb. Addition of monocytes to cultures of T cells and OKT3 hybridomas resulted in suppression of IL-2 production in a concentration-dependent manner, suggesting that monocytes regulate the levels of IL-2 production. This monocyte-independent system may be useful for further dissection of T-cell activation and proliferation and its regulation by monocytes. PMID- 2969777 TI - Flow cytometric analysis of cell-surface binding elements for fibronectin on mouse lung cell isolates. AB - Fresh lung cell isolates from LAF1 mice were examined for the presence of fibronectin-binding elements using flow cytometric analysis. Thoroughly perfused lungs from adult male mice were dissociated using an elastase-trypsin digestion, gentle pipetting and filtering. The resulting heterogeneous cell suspension was incubated with fibronectin coated 0.5 micron fluorescent beads. Subsequent flow cytometric analysis indicated the presence of two species of specific fibronectin binding populations; one of higher binding affinity which can be blocked with exogenous plasma fibronectin and one of lower binding affinity. We tentatively identify the lower affinity binding element with the fibronectin adhesion receptor and the higher affinity element with the putative matrix assembly receptor. PMID- 2969778 TI - Human lipocortins not similar to ras gene product. PMID- 2969779 TI - Dorsopathies. AB - This chapter has sought to address three aspects of back pain: the size of the problem, risk factors and preventive measures. In relation to each aspect the epidemiological approach, with particular reference to sociological and environmental factors, has been shown to have a useful role in clarifying what is on the whole a woolly clinical field. Furthermore, the problems of back pain present a challenge to clinicians from the many different specialties, including rheumatologists, orthopaedic surgeons, gynaecologists and psychiatrists, as well as those practising in more general fields as physicians and surgeons. In the face of such diversity of skills it is unlikely that a consensus will be reached without resorting to basic epidemiological principles. Back pain is among the most important cause of absence from work and long-term disability in Britain. Not only is there a formidable cost in terms of lost earnings, but those affected make heavy demands on the medical services and social security. The extent to which occupational hazards contribute to the aetiology by accelerating the onset of degenerative changes in the musculoskeletal system is difficult to assess. More studies need to be made, particularly among those who retire prematurely, change their jobs or stay off work for long periods. Cohort studies carried out prospectively are costly in manpower and time but it is only by such measures that light may be shed on factors about the workers, their low back pain and the tasks required by particular jobs which could affect prognosis. Primary prevention by controlling the weight and bulk of material being handled and also the posture adopted at work could be a possible starting point in preventing low back pain. Here again, more work needs to be done to establish which tasks and methods of performance are particularly hazardous. As far as secondary prevention or the early identification of those at risk is concerned, the low specificity and sensitivity of screening tests cast doubt over their effectiveness at the present time. There remain, therefore, many patients requiring rehabilitation, and this problem seems unlikely to decrease. Closer liaison needs to be established between health authorities and industry--possibly by such methods as attaching Disablement Resettlement Officers to hospital and appointing specialists in rehabilitation whose responsibility is not restricted to hospital based patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969780 TI - Relationship of histidine sensitivity to DNA damage and stress induced responses in mutagen sensitive mutants of Neurospora crassa. AB - Previous work in other laboratories has shown that several mutagen sensitive mutants of Neurospora crassa are extremely sensitive to low levels of histidine in the culture medium. We have shown that wild type Neurospora accumulates nicks or breaks in the DNA in the presence of histidine. The number of nicks accumulating in histidine sensitive mutants is found to increase in relation to their sensitivity to histidine. Although these nicks can be repaired by both wild type and histidine sensitive mutants when histidine is removed from the medium, a steady state number of nicks exists as long as histidine is present. We suggest that the presence of these nicks or breaks induces an increase in recombination in these possibly recombination defective mutants and that this is the source of the high level of histidine sensitivity. We speculate on the mechanisms by which histidine induces this DNA damage. This report also shows that several polypeptides are induced by the wild type organism in the presence of histidine. Some of these polypeptides are also induced during other stress situations, such as heat shock and DNA damage due to ultraviolet irradiation. Two of the histidine induced proteins cannot be induced by any of the histidine sensitive mutants. PMID- 2969781 TI - A Neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger RNA efficiently, without preference for heat shock messages. AB - Cell-free protein synthesis systems were prepared from normally-grown (N-lysate) and heat-shocked (HS-lysate) Neurospora crassa mycelium. Although both lysates translated homologous mRNA, the HS-lysate was more active, yielding a higher incorporation of [35S]-methionine into hot TCA-insoluble material and a vastly superior protein synthesis profile. The optimal temperature for translation by both lysates was 21 degrees C; the HS-lysate did not translate heat-shock mRNA preferentially at any temperature tested. Fortuitously, heterologous messenger RNAs from diverse eukaryotic and viral sources - Drosophila, dog pancreas, rabbit globin mRNA, brome mosaic virus, tobacco mosaic virus - were translated by the HS lysate with an efficiency comparable to that of the commercial rabbit reticulocyte system and superior to the wheat germ system. The cap analogues, m7G(5')ppp(5')G and m7G(5')Gm, inhibited translation significantly. PMID- 2969783 TI - Sex-dependent, tissue-specific opposing effects of dehydroepiandrosterone on initiation and modulation stages of liver and lung carcinogenesis induced by dihydroxy-di-n-propylnitrosamine in F344 rats. AB - Administration of the hormone dehydroepiandrosterone (DHEA) (0.6% in the diet) during or subsequent to injections of the carcinogen dihydroxy-di-n propylnitrosamine (DHPN) (2 X 1000 mg/kg body weight, i.p.) brought about alteration in the yield of preneoplastic lesions in liver and lung of both male and female F344 rats. Concomitant treatment with DHEA was associated with decrease in the numbers and size of glutathione S-transferase (GST-P)-positive hepatocellular foci while effecting a significant increase in development of lung lesions, especially in females. Long-term treatment with the hormone subsequent to carcinogen exposure brought about a reduction in numbers of liver foci in both sexes but in males was also associated with the development of large GST-P negative foci and nodules of amphophilic/tigroid cell character. DHEA itself did not induce any focal lesions in the lungs or livers of either sex. Thus the hormone increased sensitivity to 'initiation' in the lung while decreasing that in the liver and exerted a sex-dependent pronounced modulation of the phenotype of a proportion of hepatocellular lesions. PMID- 2969782 TI - Analysis of cDNA clones encoding the entire precursor-polypeptide for ferredoxin:NADP+ oxidoreductase from spinach. AB - In this paper, we report the structural characterization of several spinach ferredoxin-NADP+ oxidoreductase (FNR) cDNAs ranging in size from 0.9 to 1.5 kilobases. A comparison of the deduced amino acid sequence with the known amino acid sequence determined for the spinach protein establishes that 1.4-1.5 kpb inserts span the full length of the mature protein (314 amino acid residues; Mr = 35,382). These also include an N-terminal 55 amino acid transit peptide as well as maximally 171 and 214 nucleotide 5' and 3' untranslated sequences, respectively. Evidence has been obtained that various forms of FNR arise from at least two similar genes. The FNR precursor (369 amino acid residues) has a calculated molecular mass of 41.2 kDa. Comparison of the transit peptide with transit peptides from two other stromal proteins shows little similarity at the level of primary sequence but some common features in secondary structure predictions. PMID- 2969784 TI - Reactivity of T-cells from patients with rheumatoid arthritis to anti-CD3 antibody. AB - The ability of an anti-CD3 monoclonal antibody (OKT3) to induce proliferation was examined in peripheral blood mononuclear cells (PBM) from 30 patients with rheumatoid arthritis (RA). Controls consisted of 10 patients with osteoarthritis, 12 patients with psoriatic arthritis, and 12 healthy subjects. The results revealed enhanced PBM reactivity in patients with active RA relative to inactive RA patients and all control groups. PBM of patients with mild/moderate clinical disease activity exhibited augmented anti-CD3 reactivity while those with severe disease demonstrated impaired reactivity. Enhanced reactivity was also observed in the active RA group using another anti-CD3 monoclonal antibody (Leu-4). Differences in anti-CD3 dose-response or time kinetics could not account for the results. Studies of enriched T-cell preparations revealed a markedly enhanced anti-CD3 reactivity of RA T-cells relative to normal control T-cells. Monocyte/T cell mixing experiments revealed no enhanced reactivity of RA monocytes in the anti-CD3 response. RA T-cell preparations depleted of monocytes by limiting dilution reacted significantly more to anti-CD3 in the presence of IL-2 relative to controls. The enhanced reactivity could be accounted for in part by hyperreactivity of the OKT8-bearing subpopulation of T-cells. PMID- 2969785 TI - Characterization of the human interleukin 1 receptor on human polymorphonuclear leukocytes. AB - Interleukin 1 (IL-1) is a mediator of inflammation with multiple proinflammatory and immunologic enhancing activities. Human polymorphonuclear leukocytes (PMN) also play a major role in the inflammatory response. We have found that PMN possess a single type of high affinity receptor for human recombinant (r) IL-1 alpha with an apparent dissociation constant of 0.28 nM. Approximately 700 receptors are present per cell. Binding is rapid with 50% of maximal binding occurring within 20 min at 4 degrees C. Internalization of the receptor occurs within 25 min after shifting the cells to 37 degrees C. The receptor exhibits an apparent molecular weight of approximately 60-70 kDa. Electron microscopic autoradiography studies reveal that the 125I-rIL-1 alpha localized in the nucleus within 180 min after shifting cells to 37 degrees C. The accumulation of relatively high levels of 125I-rIL-1 alpha in the nucleus is consistent with earlier observations on the nuclear localization of IL-1 in T lymphocytes. The possibility that IL-1 may exert a direct action in the nucleus remains to be determined. PMID- 2969786 TI - Immunological mechanisms of corticosteroid therapy in chronic active hepatitis: analysis of peripheral blood suppressor T-cell and interleukin 2 activities. AB - To investigate the immunological mechanisms underlying corticosteroid therapy in chronic active hepatitis (CAH), in vitro effects of prednisolone on suppressor T cell and interleukin 2 (IL-2) activities were examined in six corticosteroid therapy-effective and six therapy-ineffective patients with CAH prior to the therapy. Whereas low suppressor T-cell activity and decreased response to IL-2 in T cells were found in the corticosteroid therapy-effective group, these reductions recovered to the normal range when the activity or response was tested in the presence of prednisolone (1 and 10 micrograms/ml). Corresponding with these recoveries, suppressor T-cell activity arrived at normal values after corticosteroid therapy for 8 weeks. By contrast, in the corticosteroid ineffective group, no apparent effects of prednisolone on suppressor T-cell activity and the response to IL-2 were observed. The relationship between the clinical effect of corticosteroid therapy and in vitro improvement in suppressor T-cell activity or in the response to IL-2 by prednisolone suggests that, in CAH, the corticosteroid effect is likely to be due to an immunomodulation in T-cell function. PMID- 2969787 TI - Recurrence of acute rejection in the absence of CD3-positive lymphocytes. AB - Allograft rejection is the single largest impediment to successful transplantation. Therapy targeted to lymphocytes has been in practice for many years using polyclonal heteroantisera. These products are generally accepted as being useful for the prevention and treatment of rejection; however, there have been problems with specificity, lot to lot variability, and supply. Therapy with monoclonal antibodies such as OKT3 may circumvent these problems and may allow for refined specificity. OKT3 has been shown to be highly effective at reversing acute renal allograft rejection. The few treatment failures were attributed to anti-mouse antibodies eliminating the OKT3, or to delay of therapy to such a late stage that rejection was irreversible. We present two cases which demonstrate successful reversal of acute rejection in cadaveric renal transplants by OKT3. The reversal was transient, however, in both cases, as both patients experienced recurrence of rejection while still receiving the monoclonal antibody. This occurred despite the absence of CD3-positive cells in the peripheral blood, and the presence of excess OKT3 in the serum. This implies that CD3-negative lymphocytes may under certain circumstances contribute to the rejection phenomenon. PMID- 2969788 TI - Physical fitness and chronic low back pain. An analysis of the relationships among fitness, functional limitations, and depression. AB - The purpose of this study was to describe the associations between physical fitness and important aspects of chronic low back pain problems, specifically, pain, depression, physical dysfunction, and psychologic dysfunction. Ninety-six persons with chronic low back pain were evaluated with a battery of physical and psychologic disability measures and basic physical fitness tests for aerobic capacity, strength, and flexibility. Greater overall physical fitness was significantly correlated with less physical dysfunction (R = -0.48) and fewer depressive symptoms (multiple R = -0.42), but not with psychologic dysfunction (R = -0.20) or pain (R = -0.20). Fitness accounted for 23% of the variance in physical dysfunction and 17% of the variance in depression in this mildly dysfunctional chronic back pain patient sample. Strength, as compared with aerobic capacity or flexibility, was the fitness variable contributing most to these observed associations. PMID- 2969789 TI - Different small nuclear ribonucleoprotein particles are involved in different steps of splicing complex formation. PMID- 2969790 TI - Current status of coronary angioplasty. PMID- 2969791 TI - Atrial natriuretic peptide decrease during spontaneous breathing with continuous positive airway pressure in volume-expanded healthy volunteers. AB - We examined the effect of spontaneous breathing with continuous positive airway pressure (CPAP) on the plasma concentrations of immunoreactive (ir) alpha-atrial natriuretic peptide (ANP). In three experiments, each of 11 healthy male volunteers performed CPAP at 20, 10 and 0 cm H2O for 2 h during continuous volume loading. Samples were drawn from a peripheral vein. Plasma concentrations of irANP were determined by a sensitive radioimmunoassay. Significantly lower concentrations of irANP were observed during 20 cm H2O CPAP than at 10 and 0 cm H2O. The concentrations of irANP did not differ significantly when individuals breathing with CPAP at 10 and at 0 cm H2O were compared. Our data suggest that CPAP at 20 cm H2O lowers the release of ANP in volume-expanded subjects. We hypothesize that this phenomenon may contribute to the fluid retention and renal dysfunction observed frequently during high CPAP levels. The decline in plasma concentrations of irANP may be the result of atrial compression by the distended lungs and of reduced venous return to the heart during CPAP. PMID- 2969792 TI - Flow rate variability from electronic infusion devices. AB - During continuous drug administration to pediatric patients, unfavorable pharmacologic effects have occurred. These effects were attributed to variations in flow from electronic infusion devices (EIDs). The intent of this investigation was to evaluate the influence of microrate (0.1 to 99.9 ml/h) EID on the accuracy, continuity, and pattern of flow of continuously effused fluid. Using a factorial study design, iv fluid was effused through iv delivery systems using combinations of five microrate EIDs, three iv flow rates, and three sample collection intervals. Serial weights were measured at the appropriate sample collection time using a computerized gravimetric technique to determine accuracy, continuity, and pattern of flow. All the EIDs produced accurate flow within 5% of the desired rate of 5 and 10 ml/h. At 1 ml/h, the actual iv flow rate ranged from 65.1% to 91% of the desired rate. Each of the respective EIDs produced various levels of flow continuity; each flow pattern characterized the mechanism of pump operation for each device. Thus, alteration in response (e.g., increased toxicity or decreased efficacy) to a continuous drug infusion must not be attributed exclusively to the drug or clinical condition of the patient. Serious consideration should also be given to the method of drug delivery and, in particular, the continuity of flow that results from a particular EID. PMID- 2969793 TI - Pancreatic islets generate allospecific cytolytic T lymphocytes in a mixed lymphocyte-islet culture. PMID- 2969794 TI - [Use of a hematoporphyrin derivative as a sensitizer to radiotherapy in the treatment of oral and maxillofacial malignant tumors]. PMID- 2969795 TI - [Plasma levels of beta-thromboglobulin and platelet factor 4 in cerebral infarction]. PMID- 2969796 TI - Selective insulin action on skin, ovary, and heart in insulin-resistant states. AB - States of hyperinsulinemia with resistance to insulin action on glucose disposal are frequently associated with proliferative tissue abnormalities of the skin (acanthosis nigricans), ovary, and heart. That insulin may be involved in the pathogenesis of these growth-related abnormalities despite resistance to its metabolic effects mediated through the insulin receptor is suggested by the known ability of high concentrations of insulin to stimulate DNA synthesis and cell proliferation in vitro through the insulin-like growth factor I (IGF-I) receptor. IGF-I receptors are present in skin keratinocytes, some ovarian tissue compartments, and in the heart. Furthermore, ovarian tissue from hyperinsulinemic insulin-resistant women responds to supraphysiologic insulin concentrations in vitro by enhanced steroidogenesis. Cultured, transformed T-lymphocytes from an infant with leprechaunism fail to augment basal-colony formation in response to physiologic insulin concentrations in vitro (compared to a doubling seen in normal subjects), but respond normally to supraphysiologic insulin concentrations, the effect of which is competitively inhibited by a monoclonal antibody to the IGF-I receptor. Thus, insulin action mediated through the IGF-I receptor may initiate growth-promoting tissue effects in the face of limited insulin effect on glucose metabolism. Such spillover actions may add to the morbidity associated with states of clinical insulin resistance. PMID- 2969797 TI - [Paranoid psychoses in HIV infection]. AB - Paranoid psychosis developed in two homosexual patients (aged 36 and 37 years) three and four years, respectively, after found to have an HIV infection. There was a marked reduction in the T-helper to T-suppressor-cell proportion (0.4 and 0.2). Neither computed tomography nor magnetic resonance imaging, nor microscopic, microbiological and biochemical tests of cerebrospinal fluid demonstrated opportunistic infection of the central nervous system. After treatment with haloperidol (3 mg daily) there was a rapid regression of the psychotic symptoms. PMID- 2969798 TI - Effects of different antihypertensive drugs on left ventricular function. AB - Left ventricular hypertrophy (LVH) is a response by the heart to haemodynamic overload. It is frequently observed in hypertension as a consequence of work overload secondary to an increased systemic resistance. This increase is not the only cause of LVH; there are other factors which can have a significant effect on its incidence. LVH in arterial hypertension acts initially as a useful compensatory mechanism against increased peripheral resistance. However after a certain amount of time it produces changes of variable intensity which have important consequences for the heart and some of them impair cardiac performance: 1. It affects both systolic and diastolic ventricular function; 2. It reduces coronary reserve; 3. It increases the incidence of angina and heart failure; 4. It increases the incidence of ventricular arrhythmias and sudden death. It therefore seems reasonable to include the reduction of LVH among the basic aims of antihypertensive treatment. Antisympathetic drugs (methyldopa, prazosin, urapidil), adrenergic beta-blockers, calcium antagonists and the converting enzyme inhibitors have proven to have a variable degree of efficacy in effecting a regression of LVH. PMID- 2969799 TI - Comparison between the effects of urapidil and methyldopa on left ventricular hypertrophy and haemodynamics in humans. AB - In a randomised double-blind study the effects on left ventricular mass (LV mass) and cardiac haemodynamics of urapidil, an antihypertensive agent with a vascular postsynaptic alpha 1-blocking action and a central antihypertensive effect, were compared with those of methyldopa in 29 patients with essential hypertension. During a 3-month period, urapidil was initially given at 120 mg/day and increased to 180 mg/day if a satisfactory antihypertensive response was not achieved. Methyldopa was started at 100 mg/day and increased to 1500 mg/day if an adequate blood pressure response was not achieved. Echocardiographic measurements were obtained at baseline and after 12 weeks' active treatment. The frequency rates of responders (DBP less than 95 mm Hg) on urapidil and methyldopa were 54% and 62%, respectively, after 12 weeks. In the group as a whole there was a nonsignificant tendency for decreased LV mass on both active drugs. However, the haemodynamic changes were difficult to interpret because of baseline differences between the 2 treatment groups. PMID- 2969800 TI - The hydrolysis of estrone sulfate and dehydroepiandrosterone sulfate by MCF-7 human breast cancer cells. AB - Reports of estrone (E1) and dehydroepiandrosterone (DHEA) sulfatase (sulfohydrolase) activities within many human breast cancers have prompted us to undertake the identification and partial characterization of these enzyme activities within MCF-7 human breast cancer cells. Enzyme assays were performed within subcellular preparations and intact cultures by quantifying the total nonpolar 3H-labeled metabolites formed from [3H]E1 sulfate (E1S) and [3H]DHEA sulfate (DHEAS). The results have shown that the hydrolysis of each steroid sulfate is mediated by different particulate enzymes, which demonstrate optimal activity between pH 6.0-7.0. The analysis of enzyme kinetic data showed the Km values of E1S and DHEAS for their enzymes to be approximately 6.3 and 3.6 microM/L, respectively. Neither enzyme was subject to product inhibition. Androsterone sulfate and pregnenolone sulfate produced significant inhibition of E1, but not DHEA, sulfatase activity. E1S inhibited DHEA sulfatase competitively, with an approximate Ki of 11 microM, whereas DHEAS inhibited E2 sulfatase in a noncompetitive fashion, demonstrating an approximate Ki of 0.6 microM. Studies carried out with intact MCF-7 cultures using physiological concentrations of 3H labeled E1S (2 nM) or DHEAS (1 microM) showed the accumulation of nonpolar metabolites during a 20-h incubation period. When cultures were incubated with similar concentrations of both steroid sulfates the apparent intracellular activity of E1 sulfatase was reduced by approximately 70%, whereas DHEA sulfatase activity remained unchanged. The results of these studies confirm the ability of MCF-7 cells to hydrolyze extracellular E1S and DHEAS, indicate that these reactions are mediated by different enzymes, and demonstrate that DHEAS is a potent inhibitor of MCF-7 E1 sulfatase. Circulating DHEAS, therefore, may substantially limit the ability of most postmenopausal breast cancers to use E1S as a substrate for intracellular estrogen biosynthesis. PMID- 2969801 TI - A monoclonal antibody to the type 1 insulin-like growth factor and insulin receptors stimulates deoxyribonucleic acid synthesis in human and murine fibroblasts. AB - Insulin-like growth factor I (IGF-I) and insulin are polypeptide hormones that stimulate their cellular responses by binding to specific cell membrane receptors. These receptors, while chemically distinct, have similar structural and functional characteristics. This manuscript describes the production and characterization of a monoclonal antibody that binds to both type I IGF and insulin receptors. This antibody did not inhibit hormone binding to either receptor type, but stimulated DNA synthesis in both human and murine fibroblasts. Ten BALB/c-BYJ mice were immunized with human placental membrane fragments, and their splenic lymphocytes were fused with SP2 AG0 mouse myeloma cells. Of approximately 3000 hybridoma clones thus obtained, 1 viable clone, designated V3,8 D7, was found to produce an antibody directed against the type I IGF receptor. Solubilized radiolabeled placental membranes immunoprecipitated with affinity-purified antibody and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions revealed bands with relative molecular masses corresponding to the nonreduced intact receptor (approximately 350 x 10(3], the alpha-subunit (130-140 x 10(3], and the beta-subunit (90 x 10(3] of the type I IGF receptor. Clonal supernatant and affinity-purified antibody precipitated solubilized receptors affinity labeled with [125I]IGF-I. Antibody V3,8 D7 also precipitated solubilized placental membranes affinity labeled with [125I]insulin. However, solubilized receptors affinity purified by the monoclonal antibody bound IGF-I much better than insulin, suggesting that this antibody has a higher affinity for the type I IGF receptor than for the insulin receptor. Affinity-purified antibody did not inhibit the binding of IGF-I or insulin to receptors on human placental membranes, suggesting that it is directed against a site on the type I IGF and insulin receptor not involved in hormone binding. However, affinity-purified monoclonal antibody stimulated DNA synthesis in human GM 498 and murine BALB/c-3T3 clone A 31 fibroblasts, as determined by [3H]thymidine incorporation. The combination of IGF-I and affinity-purified antibody did not increase thymidine incorporation above levels observed with either substrate alone, suggesting that these factors may be operating through a common mechanism. These results suggest that antibody V3,8 D7 can stimulate receptor responses by binding to a site on the type I IGF and/or insulin receptors that is not involved in hormone binding. These data support the concept that hormone receptors themselves possess the biological information required for stimulating specific cellular responses. PMID- 2969803 TI - On the bending moment capability of the pressurized abdominal cavity during human lifting activity. PMID- 2969802 TI - Androgenic activity of dehydroepiandrosterone and androstenedione in the rat ventral prostate. AB - In order to assess the androgenic potency of physiological plasma concentrations of the adrenal steroids dehydroepiandrosterone (DHEA) and androstenedione (delta 4-dione) in the rat prostate, these two steroids were released from Silastic tubings of appropriate length and size in castrated male rats. Implants of DHEA led to plasma levels of DHEA and 5-androsten-3 beta,17 beta-diol covering the range of concentrations found in adult men while no significant change was observed in plasma levels of delta 4-dione, testosterone and dihydrotestosterone (DHT). delta 4-Dione implants, on the other hand, led to a parallel increase in plasma delta 4-dione and testosterone levels at all doses used while plasma DHT only increased at supraphysiological doses of delta 4-dione. At plasma concentrations comparable to those found in adult men; delta 4-dione (0.8 ng/ml) and DHEA (3.4 ng/ml) stimulated prostate weight 3.7- and 2.1-fold, respectively. In the same groups, prostatic DHT levels were elevated at 4.48 +/- 0.05 and 2.70 +/- 0.73 ng/g tissue, respectively. A close parallelism was observed between prostatic DHT levels and prostatic weight in all groups. The present data show that in the rat, a species having no significant secretion of adrenal androgens, plasma concentrations of DHEA and delta 4-dione maintained within the range of those found in adult men are efficiently converted into DHT and act as potent androgenic stimuli in prostatic tissue. The castrated rat bearing Silastic implants releasing constant and predetermined amounts of adrenal steroids offers a good model to study the recently identified role of adrenal steroids in peripheral tissues. PMID- 2969804 TI - Phorbol 12,13-dibutyrate and mitogens increase fructose 2,6-bisphosphate in lymphocytes. Comparison of lymphocyte and rat-liver 6-phosphofructo-2-kinase. AB - The influence of tumour promoters and growth factors on glycolysis and on fructose-2,6-bisphosphate concentration was studied in isolated mouse spleen lymphocytes and in purified B-cells. The intracellular concentration of fructose 2,6-bisphosphate and the rate of lactate release were increased 2-3-fold in spleen lymphocytes exposed to active phorbol esters, mitogenic lectins, interleukin 4 or lipopolysaccharide. The maximal effect was observed after 1 h of exposure. In these cells hexose 6-phosphates increased 2-fold and 6-phosphofructo 2-kinase activity remained unchanged after treatment with phorbol 12,13 dibutyrate or with lectins. Exposure of B-cells to phorbol 12,13-dibutyrate, interleukin 4 or lipopolysaccharide increased the glycolytic flux and the concentration of fructose 2,6-bisphosphate without relation to their mitogenic activity. Lymphocytes and rat liver 6-phosphofructo-2-kinase were partially purified using the same procedure. The lymphocyte enzyme was not inhibited by sn glycerol 3-phosphate in contrast to the potent inhibition observed in liver. Treatment of both enzymes with the catalytic subunit of the cyclic-AMP-dependent protein kinase failed to inactivate 6-phosphofructo-2-kinase from lymphocytes. These differences suggest that lymphocytes and liver contain different forms of this enzyme. PMID- 2969805 TI - Striated muscle overload. AB - In response to increasing demand, cardiac muscle develops several adaptational mechanisms. Gene expression is modified: the heart hypertrophies and its structure changes in order to improve the efficiency of the contraction. The sarcomere modifications are both species and tissue specific. An isoenzymic shift of myosin from the high ATPase activity form V1 to the slow activity form V3 occurs in all conditions where V1 is initially predominant, i.e. rat (and also rabbit) ventricles and the atria of other species, including humans. The isoenzymic shift was not observed in conditions where V3 is predominant, as in human (and also cat and pig) ventricles. Similar changes are observed in skeletal muscle suggesting that the primary determinant of these modifications is not dependent on the innervation but only on the mechanical activity. PMID- 2969806 TI - Surgery of aortic stenosis and lesions of the ascending aorta. AB - Between 1978 and 1986, 442 aortic valvular replacements were performed for chronic calcified aortic stenosis. In 11 patients (2.5%, seven men and four women) replacement of the ascending aorta was associated with the valvular replacement, because of: seven supra-coronary aneurysms, two aneurysms involving the Valsalva sinuses, and two acute dissections of the ascending aorta. The repair of the ascending aorta consisted in inserting: one patch on one Valsalva sinus, seven supra coronary Dacron prostheses, three valved conduits (according to the Bentall technique). One patient died postoperatively (9%). PMID- 2969808 TI - Fluid mechanics of aortic stenosis. AB - In vitro qualitative and quantitative flow-mapping studies were conducted in an adult size aortic flow chamber, using bioprosthetic valves (0.5-5.0 cm2) to mimic varying degrees of aortic stenosis. The studies were performed under physiologic conditions in a left heart stimulator using: flow visualization, laser Doppler anemometry (LDA), continuous-wave (CW) Doppler and colour Doppler flow mapping (CDFM) techniques. Pressure gradients in the range 15-150 mmHg were accurately predicted by CW Doppler using the Bernoulli equation (r = 0.99). The flow visualization and CDFM studies revealed that all degrees of aortic stenosis led to jet-type flow fields, in which jet orientation was not necessarily symmetric and was skewed to varying degrees. Therefore, in aortic stenosis, Doppler measurements should be conducted in multiple views in order to visualize the flow field properly. Measurements with cross-sectional LDA revealed that as aortic stenosis increased: jet size narrowed; the peak velocity and turbulent intensities of the jet increased; jet instability increased; and acceleration of the jet proximal to the valve increased. Peak velocities as high as 4-7 ms-1 with turbulence levels (i.e. root mean square axial velocities) of 1.0-2.3 m s-1 were measured, with the moderately and severely stenotic valves. These elevated levels of turbulence could cause damage to the formed elements of blood and the walls of the ascending aorta. The high velocities and turbulence levels created by the moderately and severely stenotic valves, made quantitative interpretation of CDFM recordings very difficult, if not impossible. PMID- 2969807 TI - Prognosis and long-term results of surgically treated aortic stenosis. AB - A total of 675 patients (mean age 60 +/- 11 years, range 22-85, 71% males) with calcified pure aortic stenosis (490) or mixed lesions (185) had an aortic prosthesis: 290 Starr-Edwards, 147 Bjork, 18 other mechanical valves, 163 pericardial xenografts, 57 porcine xenografts; 76 patients simultaneously underwent a coronary bypass, 12 a replacement of ascending aorta, and six a mitral valvuloplasty. Preoperatively, 67% were in functional class III or IV (NYHA); 69.5% had a coronary arteriography: significant coronary stenosis was observed in 27% of patients. The operative mortality was 6.8% depending on the functional class at surgery, age (4.9% before 70 years, vs. 12.5% after), and the date of surgery (8.1% before 1983 vs. 3.7% after). The 10- and 15-year actuarial survival rates were 62 +/- 3% and 44 +/- 4%, respectively. No significant differences were observed between patients with pure aortic stenosis and mixed aortic lesions. In patients over 70 years, the survival rates were 71 +/- 5% at five years and 51 +/- 10% at 10 years. Age, functional class, degree of congestive heart failure, and degree of cardiomegaly were the main preoperative predictors of late death. At 10 years, 88% of patients were free from myocardial dysfunction, 87% from thromboembolic events, and 89% from haemorrhages. We conclude that in this type of valvulopathy, short- and long-term surgical results are good despite the fact that most patients are elderly and in an advanced functional class. PMID- 2969809 TI - Percutaneous transluminal aortic valvuloplasty: indications and results in adult aortic stenosis. PMID- 2969810 TI - Treatment of calcified aortic stenosis: surgery or percutaneous transluminal aortic valvuloplasty? AB - A total of 546 patients with severe aortic stenosis (AS) were retrospectively reviewed to assess the efficacy of percutaneous transluminal valvuloplasty (PTV) and valve replacement (VR). Of these, 490 underwent VR between 1968 and 1986 (mean age 62 +/- 12 years, 71.7% were in NYHA class III or IV), 68.8% received mechanical prostheses, and in 11.8% a bypass graft was associated. The operative mortality was 6.9% (4% since 1983). The mean follow-up was 57.6 months. The actuarial survival rate was 77% at five years, 60% at 10 years, and 40% at 15 years. Over 70 years of age, operative mortality was 6.2% since 1983, and the actuarial survival rate was 67.5% at five years. From February 1986 to May 1987, PTV was attempted in 56 patients and was effective in 52 patients (mean age 79 +/ 5 years, 93% in Class III or IV). Immediate mortality was 7.1%. The morbidity was due to tamponade (1.8%), myocardial infarction (3.6%), vascular trauma (5.3%), or cerebrovascular accident (9%). Forty three patients were followed after PTV (mean value 4.2 +/- 3, range 1-14 months): 12 patients (28%) died and 46% were functionally improved in NYHA Class II. PTV significantly improved the aortic valve area as shown by haemodynamics (0.49-0.75 cm2; P less than 0.0001) and these findings were corroborated by Doppler study (0.46-0.70 cm2, P less than 0.001). In conclusion, this series shows that surgery provides satisfactory results in AS with a low mortality and good long-term results, even in the elderly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969811 TI - Left ventricular systolic function in aortic stenosis. AB - In aortic valve stenosis, concentric hypertrophy develops which is characterized by a reduced end-diastolic radius-to-wall thickness ratio (r/h) with an essentially normal cavity shape. As long as the product of (r/h) and LV systolic pressure remains constant, hypertrophy is appropriate. An increase in the product, which represents an increase in wall stress signals inadequate LV hypertrophy. Although at first glance, massive LV hypertrophy appears favourable for the maintenance of a normal LV ejection fraction in aortic stenosis, data from 23 studies of the literature have shown an inverse relationship between ejection fraction and LV angiographic mass m-2 (r = -0.59). Both a degree of hypertrophy inadequate to keep systolic wall stress within normal limits and a reduction of LV contractility may explain the depression of ejection fraction when LV angiographic mass is sizeably increased. Conversely, a normal ejection fraction in aortic stenosis may not be indicative of normal systolic myocardial function under all circumstances. In the presence of mildly reduced contractility, a normal ejection fraction may be maintained by the use of preload reserve. Assessment of myocardial structure from LV endomyocardial biopsies revealed no differences in muscle fibre diameter, interstitial fibrosis and volume fraction of myofibrils between patients with aortic stenosis having a normal and those with a depressed ejection fraction. Preoperative ejection fraction is a poor predictor of postoperative survival, whereas markedly increased preoperative angiographic mass and end-systolic volume have been reported to predict an unsatisfactory postoperative outcome characterized by either death or poor LV function. PMID- 2969812 TI - Mechanical adaptation to chronic pressure overload. AB - According to Meerson, the adaptation to cardiac overload can be divided into three periods: the first stage, immediately after the initiation of the defect during which hypertrophy develops, followed by the stable hypertrophy phase (SHP), and a third phase of myocardial failure. Ventricular muscle contraction during SHP has been extensively studied both in vivo and in vitro with conflicting results. In isolated papillary muscles, most studies showed a normal or depressed contractility during chronic volume overload and a depressed inotropic state in pressure overload with a reduced maximal velocity of shortening which has been related to a myosin isozyme shift. In contrast, in conscious animals, haemodynamic status is usually described as preserved during SHP with a ventricular hyperfunction and a normal contractile function per unit of muscle. This was the basis of the concept of preload reserve and afterload mismatch described by Ross. However, mechanisms other than preload reserve may play a role during cardiac adaptation to pressure or volume overload. For instance, we recently showed in the early phase of pressure overload an increased inotropic state of the in situ heart with a change of the excitation contraction coupling evidenced by a modification of the force-frequency relations. Changes in the adrenergic receptors (density and/or affinity) may also contribute to the adaptation of the in situ heart to cardiac overload. They represent an important research area because they may explain, along with species and model differences, the discrepancies between in vivo and in vitro studies. PMID- 2969813 TI - Doppler flow mapping and its comparison with the continuity equation method for quantifying aortic stenosis. AB - The flow-mapping technique, which detects and planimeters the area of systolic flow at the site of the aortic orifice, was applied to 59 patients with a stenosed aortic valve, all of whom underwent cardiac catheterization. The success rate was 93%. The correlation coefficient between the values of valvular areas obtained by Doppler and those yielded by the Gorlin formula was r = 0.93 (SEE = 0.12 cm2). The continuity equation procedure, with the use of the velocity-time integrals, was applied sequentially to 20 of the above mentioned patients. The success rate was 85%. The valvular areas obtained in these patients by the Gorlin formula correlated well with those obtained with flow mapping (r = 0.90, SEE = 0.14, standard deviation of the difference = 0.13 cm2), as well as with those yielded by the continuity equation procedure (r = 0.86, SEE = 0.17 cm2, standard deviation of the difference = 0.16 cm2). Furthermore, the data from both ultrasonic methods were satisfactorily cross-correlated (r = 0.92, SEE = 0.12 cm2). It is noteworthy that the values of aortic valvular area obtained by Doppler were slightly larger than those found using either the continuity equation procedure or the Gorlin formula. The authors conclude that the flow mapping technique represents a reliable method for quantifying stenotic aortic valvular area and correlates well with the continuity equation procedure. It is therefore suggested that, whenever possible, both techniques should be used sequentially as a valuable and practical cross-checking policy. PMID- 2969814 TI - Percutaneous transluminal coronary angioplasty and flying status. PMID- 2969815 TI - B cell participation in the recursive selection of T cell repertoires. AB - Normal BALB/c mice produce 2,4,6-trinitrophenyl (TNP)-I-Ad specific T helper (Th) cells expressing a receptor heterodimer which share with anti-TNP antibodies an idiotope defined by the F6(51) anti-idiotypic antibody. Expression of this Th idiotype is controlled by major histocompatibility complex and immunoglobulin heavy chain-linked genes and results from antibody-dependent selection of T cell repertoires (Martinez-A. et al., Eur. J. Immunol. 1986. 16: 417). We now present evidence for the recursive nature of T----B cell repertoire selection and suggest that perinatal B cells, present in adult peritoneal cavity, operate in the early phases of this process. Thus, the Th idiotype is absent in BALB/c mice which are either suppressed from birth with anti-mu antibodies, or reconstituted with autologous bone marrow after lethal irradiation as adults. Supplementation of bone marrow reconstitution with syngeneic Thy-1-, Ly-1+ peritoneal B cells, however, selects Th cell repertoires that are undistinguishable from normal mice as to expression of the F6(51) clonotype. This effect is lost after depletion of Ly-1+ cells in the reconstituting Thy-1- peritoneal cell population. Interestingly, large in vivo "naturally" activated Ly-1- splenic B cells can also reconstitute Th idiotype expression if they are isolated from normal, but not from athymic, nude donors. However, transfer of normal large splenic T cells to adult nude mice "educates" the splenic "large B cell" compartment in these animals such that they acquire the ability to recursively select, upon transfer to bone marrow reconstituted recipients, the Th clonotype.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969816 TI - Structure and rearrangement of the T cell receptor J alpha locus in T cells and leukemic T cell lines. AB - The vast majority of T cells express an antigen receptor (TcR) composed of an alpha/beta heterodimer. The alpha and beta chains are encoded for by a set of variable (V), joining (J) and constant (C) region genes. Unlike the J genes of the beta chain which are limited in number and are clustered close to the constant region, the J alpha genes are spread over an 85-kilobase DNA region, upstream of the C alpha gene. We have isolated the complete J alpha locus, bounded on the 5' side by the C sequence of the delta gene and on the 3' side by the C sequence of the alpha gene. The experiments described here demonstrate that the J gene segments extend 75 kb 5' of C alpha and participate equally in generating the diversity of the alpha chain in peripheral T cells. Similarly, in leukemic T cell lines, rearrangements occurred over the entire locus and involved both alleles. Densitometry data suggest that in most peripheral T cells both alleles also are rearranged; thus, allelic exclusion in the alpha locus does not occur at the level of rearrangement. In three cell lines, an identical rearrangement has occurred on one allele in a region located 10 kb from the 5' end of the locus. PMID- 2969817 TI - Functional T cell receptor delta chain gene messages in athymic nude mice. AB - The rearrangement and expression of T cell antigen receptor (TcR) delta chain genes were investigated in congenitally athymic nude mice. The lymphoid cells derived from nude mice showed evidence of rearranged delta chain genes and a relatively high level of delta chain gene messages. Nucleotide sequence analysis revealed that nude mice contained an in-frame delta chain transcript, composed of V delta 5-D delta 1-D delta 2-J delta 1-C delta genes in the spleen. We have previously described functionally rearranged TcR gamma chain genes in the spleen of nude mice. Taken together, T cell precursors appear to proliferate and differentiate along the extrathymic pathway into TcR gamma/delta-bearing T cells in nude mice. PMID- 2969818 TI - Profiles of lymphokine activities and helper function for IgE in human T cell clones. AB - A large panel of phytohemagglutinin (PHA)-induced T cell clones (690 in total), established from four different human lymphoid tissues (peripheral blood, tonsils, lymph nodes and spleens) by a high-efficiency cloning technique, was characterized according to their pattern of lymphokine production. The majority of both CD4+ and CD8+ clones from all lymphoid tissues produced interleukin (IL) 2 and/or interferon (IFN)-gamma in response to 24-h stimulation with PHA. In contrast, higher proportions of IL 4-producing clones were found among CD4+ clones from tonsils and spleens than from peripheral blood and lymph nodes, whereas only a minority of CD8+ clones from all lymphoid tissues were found to produce IL 4. It was not possible to divide the CD4+ (helper/inducer) clones on the basis of their pattern of lymphokine activity into two clear-cut groups analogous to Th1 and Th2 helper clones described in mice. Although 21 out of 503 (4%) CD4+ T cell clones produced IL 4, but not IFN-gamma or IL 2, and 208 (41%) produced IL 2 and/or IFN-gamma, but not IL 4, a total number of 185 (37%) CD4+ clones showed the ability to produce IL 4 plus IL 2 and/or IFN-gamma. All types of CD4+ T cells (as classified according to their pattern of lymphokine activity) provided help for IgG production in allogeneic B cells. In contrast, helper function for IgE was detectable only among the IL 4-producing clones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969819 TI - Rearrangement and expression of T cell receptor and immunoglobulin loci in immortalized CD4-CD8- T cell lines. AB - Injection of newborn mice with mixtures of wild-type moloney murine leukemia (Mo MuLV) virus and other recombinant retroviruses harboring the myc oncogene alone or in combination with the H-ras oncogene resulted in a 100% incidence of lymphatic leukemias from which permanent cell lines could be established in vitro. These cells are immunoglobulin (Ig)-, Thy-1+BP- and CD8-CD4- indicating that they are early thymocytes. Such transformed pre-T lines lack retroviral myc and ras genes but occasionally possess proviral insertion near to their endogenous myc and pim genes. We show that both Ig heavy chain (Igh) and T cell receptor (TcR) genes are rearranged in most of these lines. In some cases, a primary recombination was followed by a secondary rearrangement at the same locus. We show that VT gamma genes can rearrange outside of their known cluster suggesting that TcR gamma diversification in such pre-T cells may be different to that in more mature T cells. Ig D-JH recombinations may precede TcR gene recombination in these early T cell lines, and some but not all express sterile Cmu transcripts. Some of these lines express surface heterodimers that appear composed of alpha and beta chains that can be immunoprecipitated with a monoclonal anti-T3 antibody but not with the anti-V beta 8 monoclonal antibody F23.1. This established pre-T cell line represents novel biological material for the dissection of T cell development and function analogous to A-MuLV transformed pre-B cells. PMID- 2969820 TI - Division of human helper T cells into two sets on the basis of the induction of anti-tumor cytotoxicity by phorbol ester and calcium ionophore. AB - Fourteen noncytotoxic human helper T cell clones were examined for autocrine proliferative responses and cytotoxicity to tumor cells after stimulation with 12 O-tetradecanoylphorbol 13-acetate (TPA) and ionomycin (Io). Although all clones responded to alloantigen, they could be divided into two groups based on their proliferative response or lack of it to TPA/Io. Nonresponders could not be converted to responder status by addition of interleukin (IL) 1 or indomethacin to the cultures. Responder status did not correlate with any of the following properties of the clones: originating donor, recognitive specificity, B cell helper activity, proliferative response to IL 2 or 4, lymphokine secretory capacity or density of expression of antigen receptors, CD4 or HLA class II molecules. Responder status did, however, correlate with the ability of TPA/Io to induce major histocompatibility complex-unrestricted cytolytic activity directed towards natural killer-resistant tumor cells. These results divide human helper cells into two types on the basis of induction of anti-tumor cytotoxicity. PMID- 2969821 TI - Atrial natriuretic factor inhibits norepinephrine release in an adrenergic clonal cell line (PC12). AB - Evidence for a presynaptic neuromodulatory effect of atrial natriuretic factor (ANF) has been obtained in rat adrenal pheochromocytoma cells (PC12), a clonal line that differentiates into adrenergic neuron-like cells when treated with nerve growth factor. ANF had no effect on basal norepinephrine (NE) release. In contrast, ANF markedly and significantly inhibited carbachol-induced NE release in a concentration-dependent manner. This result is consistent with the hypothesis that ANF may be an inhibitory neuromodulator. PMID- 2969822 TI - Antidipsogenic action of a novel peptide, 'brain natriuretic peptide', in rats. AB - The effect of intracerebroventricular (i.c.v.) administration of brain natriuretic peptide (BNP) on water drinking was studied in rats. The i.c.v. injection of BNP at a dose of 1.5 nmol elicited no apparent change in spontaneous water intake in rats but significantly attenuated the water intake induced by the i.c.v. administration of 0.1 nmol of angiotensin II. The antidipsogenic action of BNP was comparable to that of atrial natriuretic polypeptide (ANP). These findings suggest that BNP could play a role in the regulation of water intake in the central nervous system, either alone or in concert with brain ANP. PMID- 2969823 TI - Central and peripheral cardiovascular effects of the enantiomers of the calcium antagonist PN 200-110. AB - The negative inotropic effects and the central and peripheral hypotensive effects of (+) and (-) PN 200-110 were investigated in cultured chick heart cells and in spontaneously hypertensive rats, respectively. There was a large difference in negative inotropic potency between the two enantiomers in cultured chick embryo ventricular cells: the (+) enantiomer was 140 fold more potent (IC50 = 1.1 +/- 0.2 nM) than the (-) enantiomer (IC50 = 160 +/- 20 nM). (+) PN 200-110 was 10 fold more potent than (-) PN 200-110 in lowering blood pressure after intravenous injection and only three fold more potent after intra-cerebroventricular injection (i.c.v.) into pentobarbital-anaesthetized spontaneously hypertensive rats. I.c.v. administered (+) PN 200-110 (1 microgram/kg) partially antagonized the hypertensive response to i.c.v. administered BAY K 8644 (30 micrograms/kg), a calcium channel agonist, while the same dose of the (-) enantiomer did not change the i.c.v. BAY-induced increase in blood pressure. These results suggest that the dihydropyridine calcium channel antagonist, PN 200-110, may act centrally and stereoselectively at the level of the dihydropyridine receptor sites involved in the control of blood pressure in spontaneously hypertensive rats. PMID- 2969825 TI - Immune response to hepatitis B vaccine given at different injection sites and by different routes: a controlled randomized study. AB - Two-hundred-ninety-nine hospital workers were randomly allocated to receive MSD hepatitis B vaccine in different injection sites and by different routes (intramuscularly by gluteal or deltoid injections or subcutaneously in the arm) or Pasteur vaccine subcutaneously. Highest rates of seroconversion and highest mean anti-HB, titres were found in subjects given MSD vaccine by intramuscular injection into the deltoid muscle. PMID- 2969824 TI - Epidemiological characteristics of hepatitis B virus infection in Cyprus. AB - The prevalence of hepatitis B virus (HBV) infection in Cyprus was estimated in a sample of 795 blood donors, 388 armed forces recruits, 1872 hospital staff, 135 haemodialysis patients, 559 thalassaemic patients, 722 institutionalised adults, 98 mentally retarded children and 323 family contacts of HBsAg carriers. Commercially available radioimmunoassays were used for screening. Merck Sharpe and Dohme hepatitis B vaccine was used for a vaccination pilot program. Our results showed that the carrier rate of HBsAg in the blood donor and army recruit samples ranged between 0.77% and 1.01% and the prevalence of past infection between 11.1% and 13.6%. Among high risk groups the highest carrier rate was found in family contacts of HBsAg carriers (18.27%), in mentally retarded children (6.12%) and in institutionalized adult patients (5.40%). The frequency of immunes was found 28.5%, 12.2% and 33.2% respectively. The highest frequency of immunes was found in thalassaemic (82.9%) and haemodialysis (43.7%) patients. The carrier rate (2.94%) and the prevalence of immunes (22.5%) in hospital personnel was higher than in general population. Vaccination studies in a limited number of thalassaemic children and hospital personnel showed excellent acceptance of vaccination and excellent immunogenicity. We infer that HBV infection should be consider as a major public health problem in Cyprus. A National Vaccination program has to be implemented for containing its spread. PMID- 2969826 TI - Interaction between platelets and lupus anticoagulant. AB - 10 consecutive patients fulfilled the diagnostic criteria for lupus anticoagulant. 4 had concomitant systemic lupus erythematosus, 1 Waldenstrom's disease and 5 had no apparent underlying disease. Only the case with Waldenstrom's disease presented a bleeding tendency, with bleeding time greater than 20 min; the others had a history of thrombotic complications. A defect of platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid was documented in the Waldenstrom's disease case whose lupus anticoagulant was an IgM. In the others, lupus anticoagulant, identified as IgG immunoglobulins, produced no aggregation abnormalities. However, beta-thromboglobulin levels in platelets, plasma and urine were consistent with a pattern of platelet activation in all cases. IgG immunoglobulins separated from sera of 6 patients showed lupus anticoagulant activity, with no effects on platelet aggregation of normal platelet-rich plasma, but they induced secretion of beta-thromboglobulin from normal platelets. PMID- 2969827 TI - Uranyl salts as photochemical agents for cleavage of DNA and probing of protein DNA contacts. AB - Single-strand DNA nicks are induced by uranyl nitrate or uranyl acetate in combination with long-wavelength (lambda approximately 420 nm) ultraviolet irradiation. The nicks occur randomly with respect to the DNA sequence. Using the lambda-repressor/ORI operator DNA system it is shown that uranyl salts can be used to photofootprint protein contacts with the DNA backbone. PMID- 2969828 TI - Phosphorylation of brush border myosin at threonine on its 20 kDa light chains by a calmodulin-independent kinase activates its ATPase. AB - A calmodulin-independent kinase isolated from chicken intestinal brush border phosphorylates brush border myosin mainly at an apparently single threonine on its 20 kDa light chains. Phosphorylation to 1.9 mol phosphate/mol myosin activated the myosin actin-activated ATPase about 12-fold, to about 100 nmol/min per mg. Brush border myosin ATPase can thus be activated by phosphorylation either at threonine, by calmodulin-independent kinase, or at serine, by calmodulin-dependent myosin light chain kinase, as previously shown [(1987) FEBS Lett. 223, 262-266]. PMID- 2969829 TI - The hydrolysis of ATP that accompanies actin polymerization is essentially irreversible. AB - The hydrolysis of ATP that accompanies the polymerization of actin occurs on the F-actin subsequent to the addition of the G-ATP-actin subunit to the elongating filament. We now show that this ATP hydrolysis is essentially irreversible. Thus, a large decrease in free energy occurs at the cleavage step, F-ATP-actin----F-ADP Pi-actin. PMID- 2969830 TI - Purification and characterization of phospholipase A2 inhibitory proteins from pig thyroid gland. AB - A 32 kDa phospholipase A2 inhibitory protein was isolated from pig thyroid gland after calcium precipitation and fast protein liquid anion-exchange chromatography. SDS-polyacrylamide gel electrophoresis revealed the purity of the protein. The protein activity was assessed by the inhibition of pancreatic phospholipase A2 on [3H]oleic acid-labelled Escherichia coli membranes as substrate and on the prostaglandin E2 production of cultured thyroid cells. The amino acid composition and the isoelectric point were quite similar to those of endonexin previously described in other tissues or cells. The cross-reactivity of a polyclonal antibody against a 32 kDa lipocortin from human peripheral blood mononuclear cells with our thyroidal 32 kDa protein confirmed its lipocortin nature. Before the purification by fast protein liquid chromatography, the Ca2+ pellet contained lipocortin I (35 kDa and its core protein 33 kDa) identified by its cross-reactivity with a polyclonal antibody. PMID- 2969831 TI - Rat cardiac hypertrophy. Altered sodium-calcium exchange activity in sarcolemmal vesicles. AB - The sodium-calcium exchange activity has been studied in sarcolemmal vesicles isolated from rat ventricles hypertrophied by pressure overload. 4 weeks after aortic stenosis the degree of hypertrophy varied from 30 to 70%. The Na+ dependent 45Ca2+ influx and efflux were up to 50% decreased and the sensitivity to Ca2+ was 13-fold lower in vesicles from hypertrophied heart as compared to those from normal heart. However, the Na+,K+-ATPase activity, the orientation of the vesicles and the passive Ca2+ permeability were found to be similar in the two heart groups. These results indicate that the sarcolemmal Na+/Ca2+ exchange activity could be qualitatively and/or quantitatively changed in hypertrophied rat heart. PMID- 2969832 TI - [Prednisolone allergy with pustular exanthema]. PMID- 2969833 TI - [Keratosis palmoplantaris with periodontopathy (Papillon-Lefevre syndrome) and inner ear deafness]. PMID- 2969834 TI - Eosinophilic cellulitis (Wells' syndrome): ultrastructural study of a case with circulating immune complexes. AB - A 42-year-old woman was observed during 3 bouts of eosinophilic cellulitis over a 6-year-period. Skin biopsies were taken at each relapse and processed for histological, immunofluorescent and ultrastructural studies. Histologically the eosinophilic infiltrate extended to the deep dermis and the subcutaneous fat. High levels of circulating immune complexes, and complement and IgG deposits around the vessels were detected for as long as the cutaneous lesions lasted. Under the electron microscope eosinophils were numerous, half of them degranulated and some granules had a double cristal core. No injury to the vessel walls was observed. The 3 recurrences occurred respectively after lincomycin, nesdonal, acetyl salicylic acid and pholcodin ingestion and responded to sulfone and steroid therapy. PMID- 2969835 TI - GnRH agonists-antagonists--clinical applications. AB - We know that there is a wide range of clinical applications for GnRH analogues in the field of benign gynaecological disorders. As we understand more of the physiology and mechanism of these GnRH agonists, and there appear more varied and perhaps efficient delivery systems, and antagonists become available, it may be possible to develop a graded approach in suppression of the hypothalamic pituitary axis. The true potential of these agents is then yet to be fully realized. There can be no doubt they are going to influence practice dramatically over the next decade. The potency of these agents, perhaps specifically their effects of oestrogen deficiency and calcium bone metabolism, suggests that limitation of duration of use and timing of recurrent administration to individuals is likely to be necessary. PMID- 2969836 TI - Ovarian stimulation for in vitro fertilization and GIFT combining administration of gonadotropins and blockade of the pituitary with D-Trp6-LH-RH. PMID- 2969837 TI - CO2 laser laparoscopy: a ten-year experience. PMID- 2969838 TI - [Determining acid phosphatase (AcP[E.C. 3.1.3.2.]) and adenosine triphosphatase (ATPase [E.C. 3.6.1.3.]) activity in the lungs of rats following a single administration of ashes from industrial high-heat facilities]. AB - There was examined pulmonary tissue of white rats, which had been administered intratrachealy a single dose of the respirable fraction of ashes sample from 6 different power stations elektrohasting plants and hasting plants in Poland (0.2 ml suspension; 50 mg of the examined sample in 0.6 cm3 of NaCl solution). 9 months after the application of the ashes, biopsies of the left lung were taken and there was determined the activity of acid phosphatase (AcP) and adenosinetriphosphatase (ATP-ase) histoenzymatically. There was found sensitivity of these hydrolases and changes of their activity connected with chemical composition of the examined ashes. PMID- 2969839 TI - Domains for protein-protein interactions at the N and C termini of the large subunit of bacteriophage lambda terminase. AB - The large subunit of phage lambda terminase, gpA, the gene product of the phage A gene, interacts with the small subunit, gpNul, to form functional terminase. Terminase binds to lambda DNA at cosB to form a binary complex. The terminase:DNA complex binds a prohead to form a ternary complex. Ternary complex formation involves an interaction of the prohead with gpA. The amino terminus of gpA contains a functional domain for interaction with gpNul, and the carboxy-terminal 38 amino acids of gpA contain a functional domain for prohead binding. This information about the structure of gpA was obtained through the use of hybrid phages resulting from recombination between lambda and the related phage 21. lambda and 21 encode terminases that are analogous in structural organization and have ca. 60% sequence identity. In spite of these similarities, lambda and 21 terminases differ in specificity for DNA binding, subunit assembly, and prohead binding. A lambda-21 hybrid phage produces a terminase in which one of the subunits is chimeric and had recombinant specificities. In the work reported here; a new hybrid, lambda-21 hybrid 67, is characterized. lambda-21 hybrid 67 is the result of a crossover between lambda and 21 in the large subunit genes, such that the DNA from the left chromosome end is from 21, including cosB phi 21, the 1 gene, and the first 48 codons for the 2 gene. The rest of the hybrid 67 chromosome is lambda DNA, including 593 codons of the A gene. The chimeric gp2/A of hybrid 67 binds gp1 to form functional terminase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969842 TI - [Experience of the Leningrad Public Health and Medical Institute in joint work with practical public health organizations]. PMID- 2969841 TI - [Contribution of scientists at the Perm Medical Institute to the improvement of health conditions among the population of the Urals]. PMID- 2969840 TI - Experimental autoimmune uveoretinitis in rats induced by rod visual pigment: rhodopsin is more pathogenic than opsin. AB - The rod visual pigment, rhodopsin, and its illuminated form, opsin, were used to induce experimental autoimmune uveoretinitis in rats. Rhodopsin appears to be more pathogenic than opsin. A dose of 250 micrograms rhodopsin injected in Freund's complete adjuvant and pertussis adjuvant induces nongranulomatous inflammation with higher frequency, which starts earlier and is more severe than that induced by opsin. Two weeks postinjection, the mean score of rhodopsin injected animals is more than twice as high as that of opsin-injected animals. The high pathogenicity of rhodopsin appears to be related to the biochemical integrity of the protein and depends on its state of illumination. The levels of the immune responses (both cellular and humoral) measured at day 10 postinjection do not account for the pronounced difference in pathogenicity between rhodopsin and opsin. The developmental patterns of severe uveoretinitis induced by rhodopsin or opsin were histologically evaluated and appear to be similar. In both cases we observed dense mononuclear and polymorphonuclear cell infiltrations in the retina and anterior uvea. Only in the severe stages does the choroid become involved. However, rhodopsin causes more pronounced involvement of the ciliary body, pars plana, and anterior chamber. The inflammation finally results in total elimination of the photoreceptor cell layer. PMID- 2969843 TI - [The G.M. Natadze Scientific Research Institute of Health and Hygiene of the Ministry of Public Health, Georgian SSR--60th anniversary]. PMID- 2969844 TI - [Improvement in help for alcoholic workers in the hydrolysis industry]. PMID- 2969845 TI - [Dynamics of the physical development of children of preschool age in the city of Gorki]. PMID- 2969846 TI - [State of the health of school children of pubertal age in the Donetz Basin]. PMID- 2969847 TI - [Professional and official requirements for laboratory physicians at departments of communal hygiene of sanitary and epidemiologic stations]. PMID- 2969848 TI - [Sub-internship as a final stage in the training of the hygienist-epidemiologist physician]. PMID- 2969849 TI - [Use of organic wastes in soil and protection of the environment]. PMID- 2969850 TI - [Hygienic regulation of the insecticide prothiophos in reservoir waters]. PMID- 2969851 TI - [Hygienic and sanitary assessment of household polymeric materials based on acrylic monomers]. PMID- 2969852 TI - [Microvascular architecture of the all-layer free inferior abdominal wall flap]. AB - The authors dissected all layers of the inferior abdominal wall to study the possibility of free transplantation to close combined skin and mucosal defects. They perfused these flaps with a special solution of lead oxide and gelatin (Rees and Taylor 1986). After fixation in formalin the flaps were dissected in their layers and radiographs of these were taken. Networks of small vessels, which originate in the inferior epigastric artery, were found preperitoneally in the deepest compartment, in the muscular compartment and, nourished by perforators via the ventral rectus fascia, epifascially and subdermally in the subcutaneous tissue. On this basis free microvascular transplantation of a full-thickness abdominal wall flap seems to be possible to close combined mucosal and cutaneous defects. To thin this flap the subcutaneous middle third could be removed taking care to protect the epifascial and subdermal plexus of vessels. An isolated pre- and peritoneal microvascular flap also seems to be possible. PMID- 2969853 TI - [A surgical method of abdominal wall reconstruction in prune belly syndrome]. AB - The "Prune Belly"-syndrome consists of a triad of anomalies: absence or hypoplastic musculature of the abdominal wall, bilateral cryptorchidism, and dilatation of the urinary tract. A surgical method of construction of the abdominal wall in such a case with a missing rectus abdominis is reported. PMID- 2969854 TI - [The effect of early systematic orthodontic treatment in trisomy 21]. PMID- 2969855 TI - Hyperglycemia alters the beta-cell sensitivity to beta-endorphin in noninsulin dependent diabetic subjects. PMID- 2969856 TI - Nicardipine induces protection against alloxan but not streptozotocin-induced diabetes in rats. PMID- 2969857 TI - Alpha human atrial natriuretic peptide and anterior pituitary hormones secretion in men. PMID- 2969859 TI - Treatment of precocious puberty with a long-acting preparation of D-Trp6-LHRH. AB - D-Trp6-LHRH was tested in 6 girls 1-8 years old and 7 boys 2-10 years old with precocious puberty. All children had advanced bone age, breast or testis enlargement and a pubertal LH response to LHRH. 60 micrograms LHRH-A/kg body weight was given intramuscularly on days 1 and 21 and thereafter every 4 weeks for 6-21 months. In girls, breast enlargement disappeared and mean uterus size decreased within 6 months. Mean ovary length decreased from 25.0 +/- 1.9 to 16.0 +/- 2.7 (p less than 0.02). In boys, mean testis volume decreased from 8.0 +/- 1.1 to 6.7 +/- 1.4 ml (p less than 0.05) within 6 months. In both sexes, growth velocity decreased significantly and bone maturation was reduced. Plasma levels of estradiol or testosterone and FSH levels decreased significantly within 3 weeks. The LH response to LHRH was reduced to normal prepubertal values after 7 weeks. No secondary clinical or biochemical escape occurred. No side effects occurred except for transient vaginal bleeding in one girl after the first and second injection. No antibodies to LHRH-A were detected in the patients' sera. This study demonstrates the ability of a delayed release formulation of D-Trp6 LHRH to suppress pituitary and gonadal secretion and pituitary response to LHRH for as long as 2 years of therapy. This treatment appears to be more efficient in treating both clinical and biochemical abnormalities than does treatment with inhibitory steroids. Additionally the method of administration is more practical and ensures better patient compliance. PMID- 2969858 TI - Treatment of central precocious puberty with an LHRH agonist (Buserelin): effect on growth and bone maturation after three years of treatment. AB - The LHRH analog Buserelin was used to treat 27 children (21 girls, 6 boys) with central precocious puberty. Nineteen patients had idiopathic precocious puberty and 8 had organic lesions (hamartoma, hydrocephalus or suprasellar arachnoid cyst). All patients received 20 or 30 micrograms/kg/day s.c. of Buserelin, and we obtained plasma E2 less than 20 pg/ml, vaginal maturation index less than 30 in girls or plasma testosterone less than 0.3 ng/ml in boys. The mean growth rate decreased from 9.3 +/- 0.5 to 4.6 +/- 1.3 cm/year after 3 years. The velocity of skeletal maturation decreased so that the final height prediction improved by a mean value of 1.6 SD. As the follow-up increases, this study confirms that LHRHa therapy is effective and potentially improves the final height of children presenting active and severe central precocious puberty. PMID- 2969860 TI - Comparative effects of cyproterone acetate or a long-acting LHRH agonist in polycystic ovarian disease. AB - A randomized cross-over study was done to compare the therapeutic efficacy of cyproterone acetate (CPA, 50 mg/day orally) and a depot preparation of the LHRH superagonist (D-Trp6 LHRH 3 mg i.m. once a month) in 10 patients with polycystic ovarian disease (PCO). The two treatment periods were separated by 6 months. Both treatments resulted in marked clinical improvement. In response to CPA treatment, basal plasma gonadotropin, estradiol, estrone, testosterone and androstenedione levels significantly decreased. In response to D-Trp6 LHRH, both basal and stimulated gonadotropin levels were completely suppressed after 3 weeks of treatment. After initial elevation on day 2, plasma ovarian steroid levels fell into the castrate range, without any change in dehydroepiandrosterone sulfate levels. Urinary 3 alpha-androstanediol excretion decreased significantly. In patients with PCO, LHRH-A induced more complete gonadotropin inhibition than did CPA. However, following cessation of either therapy, the disease rapidly recurred. PMID- 2969861 TI - Influence of testosterone substitution on sperm suppression by LHRH agonists. AB - To investigate the effect of LHRH (GnRH) agonists on sperm suppression, we studied the effect of a depot preparation of D-Trp6 LHRH in 10 normal men for 30 weeks. In addition, to determine the role of androgenic substitution on sperm suppression, the volunteers were divided into two groups: group 1 (n = 5) received a low dose T substitution (125 mg of T enanthate every month), while group 2 (n = 5) received a normal T substitution (120 mg of T undecanoate every day). Four men became azoospermic in group 1 and none in group 2. Moreover, administration of additional T injections in 1 volunteer of group 1 resulted in the reappearance of spermatozoa in the ejaculate. Return to the low dose therapy produced azoospermia. These results suggest that testosterone supplementation supports spermatogenesis. PMID- 2969862 TI - Antiandrogens and hirsutism. AB - The clinical appearance of female idiopathic hirsutism and its pathophysiological aspects and the antiandrogen drugs in relation to the therapy of hirsutism are discussed. Two compounds have been widely employed, namely cyproterone acetate, mainly in the 'reverse sequential regimen', and spironolactone, with or without the association of a contraceptive pill containing cyproterone acetate and ethinylestradiol. The ability to compete with dihydrotestosterone in skin androgen receptors, shared by these two compounds, has led to excellent clinical results for many years. A topical antiandrogen therapy would be a further advantage for these patients. PMID- 2969863 TI - Severe hypoplasia of lymphoid tissues in Mo1 deficiency. AB - A lymph node was examined from a 4-year-old female child with documented "Mo1" (CR3) deficiency. There was hypoplasia of the lymph node, with small, poorly delineated germinal centers and overall lymphopenia. Retrospective analysis of the clinical course of a decreased elder sister of the index patient indicated that the elder sister probably also had "Mo1" deficiency. Review of findings at autopsy revealed severe hypoplasia of all lymphoid organs with lymphopenia. Our observations suggest that lymphoid tissue hypoplasia may be a feature of CR3 deficiency and is likely a result of the defective lymphocyte adherence functions to endothelium associated with absent lymphocyte function-associated antigen (LFA 1). PMID- 2969864 TI - Histologic and immunohistochemical investigation of neuroblastomas and correlation with prognosis. AB - Forty-one confirmed cases of childhood neuroblastoma diagnosed over a 13-year period were reviewed and reclassified. Most of the tumors were stained using a peroxidase antiperoxidase method for neuron specific enolase (NSE), protein gene product (PGP) 9.5, and S100 protein, all of which have previously been reported to be positive in some neuroblastomas. The relation to prognosis of the histology and immunohistochemistry was studied. There was a significant trend toward improved survival with increasing degree of differentiation, and with decreasing mitosiskaryorrhexis index (MKI) in the stroma-poor group. There was no significant correlation between immunohistochemical staining and survival, although the presence and amount of staining for all three markers tended to increase with tumor differentiation. This study concludes that histologic classification in neuroblastoma is helpful in assessing prognosis but that the clinical features are generally more reliable as indicators of prognosis. The immunohistochemistry of markers used did not contribute towards assessment of prognosis. PMID- 2969865 TI - Three-dimensional collagen matrices as cell culture substrata affect the generation of alloreactive cytotoxic T lymphocytes. AB - Primary one-way mixed lymphocyte cultures (MLC) of C3H/He responder and DBA/2 stimulator were performed in three-dimensional (3-D) collagen matrices and the generation of alloreactive cytotoxic T cell (CTL) responses was compared to those in MLC which were done on usual plastic surfaces or on collagen-coated plastic surfaces. MLC in the 3-D collagen matrices were found to generate strong CTL responses. Flow cytometric analysis of Lyt-2 and L3T4 antigen expressions on the effector cells showed that the Lyt-2/L3T4 ratios were substantially higher in the 3-D collagen matrices, and that a larger proportion of the cells in the 3-D collagen matrices were Lyt-2+ lymphoblasts. These results indicate that the milieu of the 3-D collagen matrices favors the proliferation of Lyt-2+ lymphocytes, and suggests that cell-to-matrix interactions in 3-D collagen matrices may play a regulatory role in the maturation process of alloreactive CTLs. PMID- 2969866 TI - Depression of proliferative response to fetal lymphocytes of retroplacental blood lymphocytes in human pregnancy. AB - Retroplacental blood lymphocytes (RPL) in human pregnancy were studied for proliferative response to related fetal lymphocytes and it was compared with that of peripheral blood lymphocytes (PBL) of the same donor. RPL proliferated poorly to related cord blood lymphocytes in 9 of 10 cases when compared with autologous PBL. RPL also showed lower proliferative response to unrelated adult lymphocytes in 7 of 12 cases. Proliferation of RPL was weaker to related cord blood lymphocytes than to allogeneic adult lymphocytes. These results indicate that the response of T cells at feto-maternal interface is impaired, especially to fetal antigens. PMID- 2969867 TI - Proliferative and/or cytotoxic activity of lymphocyte clones to autologous human melanoma. AB - Peripheral blood lymphocytes (PBL) of a patient with metastatic melanoma were cultured with autologous melanoma cells (Auto-Me) and recombinant interleukin 2 (IL-2) (MLTC-PBL). Thirty-five days later, when no cytotoxicity against Auto-Me or K562 was detectable, MLTC-PBL were cloned in the presence of Auto-Me, IL-2 (25 U/ml) and Daudi cells as feeder. Eighty-one growing clones were simultaneously screened for proliferative and cytotoxic activity to Auto-Me. Twenty-two clones proliferated in the presence of Auto-Me only, 29 in the presence of IL-2 only and 41 in the presence of Auto-Me plus IL-2; 12 clones showed cytotoxic activity against Auto-Me. Six clones expressed both cytotoxic and proliferative activity to Auto-Me. The phenotype of 6 proliferative clones tested was CD3+, CD4+, WT31+, CD8-, CD16-, Leu19-, whereas that of 2 cytotoxic-proliferative clones tested was CD3+, CD8+, Leu19+, WT31+, CD4-, CD16-. Specificity analysis of proliferative response of 6 clones and of cytotoxicity of 7 clones, tested on a panel of 14 different target cells, revealed a complex pattern of reactivity, each clone expressing a peculiar specificity. Our results suggest the possibility of isolating, from melanoma patients' PBL, T-cell clones with proliferative activity to Auto-Me and Auto-Me plus IL-2, and T-cell clones which apparently express both proliferative and cytotoxic activity to Auto-Me. PMID- 2969868 TI - Photosensitization of differentiating Friend erythroleukemic cells by hematoporphyrin derivative and the cholesterol effect. AB - The Friend erythroleukemia cell line was used to study the binding and biological properties of the photosensitizer hematoporphyrin derivative (HPD) on a differentiating system. In addition, the effect of cholesterol hemisuccinate (CHS) enrichment of cell membranes on HPD activity was tested on the same cell system. Differentiation of Friend erythroleukemia cells (FLC) was induced with dimethyl sulfoxide (DMSO) and resulted in a decreased cell volume and an increased rate of hemoglobin synthesis as a function of the duration of DMSO treatment. Differentiated cells seem to bind less porphyrin than their undifferentiated counterparts. Thus, cells treated for 6 days with DMSO bound 30 40% less dye than an identical number of untreated FLC. In contrast, a similar inhibition of both DNA and protein synthesis by photoactivated HPD was evident in either DMSO-treated or untreated FLC. Enrichment of cell membranes with CHS led to the same degree of protection from the damaging activity of the photoactivated dye in both differentiated and undifferentiated FLC. The decreased binding of HPD to DMSO-treated FLC is most likely a result of a reduction in cell volume of differentiated cells and is not related to an intrinsic property of the differentiation process. PMID- 2969869 TI - Vaporization of atheroma in man: the role of lasers in the era of balloon angioplasty. PMID- 2969870 TI - Percutaneous transluminal coronary angioplasty for coronary stenosis following radiotherapy. AB - A 50-year-old woman who developed severe angina pectoris 67 months following radiation therapy of the left side of the chest for adenocarcinoma of the left breast is reported. Angiographic studies showed an isolated severe stenosis in the left anterior descending coronary artery. Successful percutaneous transluminal coronary angioplasty was performed. PMID- 2969871 TI - The distribution of dracunculiasis in Nigeria: a preliminary study. AB - This paper presents a preliminary assessment of the distribution and endemicity of dracunculiasis in Nigeria. The disease is found in all 19 States of the Federation and in the Federal Capital Territory of Abuja. It occurs in areas with a dry season of more than nine months as well as those with a dry season of less than four months; the seasonal distribution of rainfall influences the peak period of disease transmission and patency. Altogether, an estimated 2.5 million cases occur every year, and at least 30% of the entire rural population is at risk from the infection. The widespread distribution of dracunculiasis throughout the country indicates the need for a national control campaign and a sustained programme of international cooperation. PMID- 2969872 TI - Guinea worm disease in Northern Uganda: a major public health problem controllable through an effective water programme. AB - A modified cluster survey was conducted in northwestern Uganda in 1984 to provide descriptive epidemiological data on dracunculiasis in a water programme target area. A total of 2014 people participated from 58 randomly selected clusters. Interviewers elicited information on age and sex of household members, number, date of emergence and location of Guinea worms, and type of and distance from water source in an endemic area. The survey yielded an incidence rate of 193 cases/1000 people per year, and a prevalence rate of 43 active cases/1000. Respondents who reported using ponds, reservoirs, valley tanks or rivers as their primary water source had the highest attack rates; those using boreholes, the lowest. Adolescents and adults differed little in risk, but the disease was less common among young children. Guinea worm disease displayed a bimodal seasonal pattern. We concluded that the survey method used for determining dracunculiasis incidence was appropriate in this setting. The incidence of this disease may be significantly reduced in Uganda through the country's commitment to the International Drinking Water Supply and Sanitation Decade. PMID- 2969873 TI - Percutaneous large bore venotomy and tract creation: comparison of sequential dilator and angioplasty balloon methods in a porcine model. Preliminary report. AB - We used a pig model to evaluate the pathologic changes that occur during large caliber percutaneous venotomy and tract creation such as is necessary to deliver the Greenfield vena caval filter. Four sequential dilator and four angioplasty balloon tract creations were performed on eight external jugular veins in four adult pigs. Gross and microscopic evaluations of the veins and surrounding tissue followed the procedures. Morphometric measurements based on extent and severity of pathologic changes in the specimens were graded from 0 (normal) to 3 (severe and/or extensive). We found that while either technique entailed damage to the veins, the sequential dilator method produced substantially more smooth muscle stretching and tearing and disruption of the internal elastica. There also was greater perivascular and intramural hemorrhage. These changes extended as far distally as the catheters and sheaths were advanced. While the number of experimental samples is too small to allow valid statistical analysis, our results suggest that the angioplasty balloon technique may be superior for tract and venotomy creation. Furthermore, large sheaths should be advanced only enough to maintain venous access to limit the extent of potential venous damage. PMID- 2969874 TI - Changes in myocardial perfusion reserve after PTCA: noninvasive assessment with positron tomography. PMID- 2969875 TI - A mouse tRNA pseudogene derived from a tRNA(Trp) coding sequence. AB - The identification and characterization of a mouse tRNA(Trp) pseudogene is reported. A synthetic oligonucleotide (31 mer), identical with the 3' half of a tRNA(Trp), was used to examine three mouse lambda clones that are known to contain clusters of tRNA genes. A fragment of one of these lambda clones strongly hybridizes to the oligonucleotide; the sequence of this region shows the presence of a gene significantly homologous to the chick tRNA(Trp). However two point mutations and a three base deletion prevent the folding of a possible transcript of this gene. The presence of a conserved promoter sequence for RNA polymerase III, that should allow the transcription of this gene, does not ensure the transcription of the gene, at least in our in vitro system. PMID- 2969876 TI - Isolation of the human glyceraldehyde-3-phosphate dehydrogenase gene by hybridization with synthetic oligonucleotides. AB - By screening a human genomic library from human lymphocyte DNA cloned in EMBL 3 vector with synthetic oligonucleotides homologous to the human GAPD cDNA 3'-non coding region as probes, a unique lambda recombinant clone (EMBL-G5) was isolated at the Tm value. Preliminary restriction analysis and sequence data proved that this recombinant clone is the human structural gene for the glyceraldehyde- 3 phosphate dehydrogenase. PMID- 2969878 TI - Recognizing occupational stress-related injuries. PMID- 2969877 TI - Chronic graft-vs.-host disease: changes in natural suppressor cells and mast cells. PMID- 2969879 TI - [Contact allergy to dexpanthenol]. AB - Eleven cases of contact allergy to dexpanthenol are reported (5 females, 6 males; mean age 62.4 years). Five patients suffered from a leg ulcer and/or stasis dermatitis. In five patients the sensitization occurred after the application of dexpanthenol-containing ointments to the face. Only one patient did not show sensitization to other common allergens. Three patients were sensitive to wool wax alcohols, which are present in a commonly used product. Dexpanthenol seems to be a rare sensitizer, yet clinically most relevant for patients with stasis dermatitis and multiple allergies. PMID- 2969880 TI - [Lupoid form of rosacea-like dermatitis]. AB - In a 9-year-old girl, a lupoid variant of rosacea-like dermatitis was observed following the topical application of potent glucocorticosteroids. In addition to the micro- and macro-papulopustular forms, this is a third clinical variant of the disease, which has to date only been described in children. PMID- 2969882 TI - Mechanics of parasternals and triangularis sterni in upright vs. supine dogs. AB - The parasternal intercostals in supine dogs are activated and shorten during inspiration, whereas the triangularis terni is activated and shortens during expiration (J. Appl. Physiol. 61: 539-544, 1986). How the two muscles respond to posture, however, is not known. Thirteen vagotomized, phrenicotomized, spontaneously breathing animals were thus studied during multiple postural changes from supine to 80 degrees head up and 20 degrees head down. Head-up tilting elicited a gradual increase in the electrical activation of both the triangularis sterni and the parasternals. Recruitment of the triangularis sterni promoted an increase in the amount of expiratory muscle shortening, but recruitment of the parasternals was invariably associated with a considerable reduction in the amount of inspiratory muscle shortening. This reduction was abolished after sectioning of the abdominal wall. We conclude that 1) the contribution of the canine triangularis sterni to rib movement increases with the assumption of the upright posture, whereas the contribution of the parasternals decreases and 2) this decrease results primarily from the load imposed on the rib cage by gravitational forces. Thus assuming the upright posture adversely affects the rib cage inspiratory muscles as well as the diaphragm. PMID- 2969881 TI - Short-term effects of testolactone compared to other treatment modalities on longitudinal growth and ovarian activity in a girl with McCune-Albright syndrome. AB - In a 6 1/2-month-old girl with McCune-Albright syndrome, gonadotropin-independent isosexual precocity and recurrent ovarian cysts, the short-term effects of surgical therapy, cyproterone acetate (120 mg/m2/d), combined medroxyprogesterone acetate (10 mg/d), and spironolactone (50-75 mg/d) treatment, and testolactone (40 mg/kg/d) were evaluated sequentially. No significant reduction of cyst frequency was achieved with any of the medical treatments. The rate of bone maturation (delta BA/delta CA) was increased and the height standard deviation score (SDS) for bone age as a potential indicator of final height was decreased with surgical treatment alone and combined medroxyprogesterone acetate and spironolactone. Both parameters normalized with cyproterone acetate and testolactone. Height velocity SDS, however, was higher with testolactone (0.97 vs. 0.45). PMID- 2969883 TI - Carbohydrate metabolism in human skeletal muscle during exercise is not regulated by G-1,6-P2. AB - Glucose 1,6-bisphosphate (G-1,6-P2) is a potent activator of phosphofructokinase (PFK) and an inhibitor of hexokinase in vitro. It has been suggested that increases in G-1,6-P2 are a main means by which PFK can achieve significant catalytic function in vivo despite falling pH and that increases in G-1,6-P2 will inhibit hexokinase in vivo. The purpose of the present study was to determine whether contraction-induced changes in flux through PFK and hexokinase are associated with changes in G-1,6-P2 in skeletal muscle. Ten men performed bicycle exercise for 10 min at 40 and 75% of maximal O2 uptake (VO2max) and to fatigue [4.8 +/- 0.6 (SE) min] at 100% VO2max. Biopsies were obtained from the quadriceps femoris muscle at rest and after each work load and analyzed for G-1,6-P2. G-1,6 P2 averaged 111 +/- 13 mumol/kg dry wt at rest and 121 +/- 16, 123 +/- 15, and 123 +/- 11 mumol/kg dry wt after the low-, moderate-, and high-intensity exercise bouts, respectively (P less than 0.05 for all means vs. rest). Flux through PFK was estimated to increase exponentially as the exercise intensity increased and muscle pH decreased at the higher work loads, whereas flux through hexokinase was estimated to increase during exercise at 40 and 75% VO2max but decrease sharply at 100% VO2max. These data demonstrate that flux through neither PFK nor hexokinase is mediated by changes in G-1,6-P2 in human skeletal muscle during short-term dynamic exercise. PMID- 2969884 TI - Accessible public transport--when will the dream become a reality? AB - This is a paper concerning the transport needs of people with disabilities. Whilst tracing to date the development of Dial-a-Ride, the door-to-door transport service for the disabled, it also examines the future of Dial-a-Ride and explores what progress is still to be achieved in the field of transport. The paper draws upon recent research undertaken by the London Dial-a-Ride Users' Association, the representative body of London Dial-a-Ride users. PMID- 2969885 TI - Transport handicap--its causes, its scale and its effect. AB - It has become clear during all stages of our research that transport plays a vital part in enabling disabled people to live independent lives and to be a full part of the community in which they live. All Change, a consumers study of public transport handicap in Greater London, published in 1986, sought to gather information about the scale, causes, and extent of transport handicap in the Greater London area. The findings are based on interviews from people who define themselves as transport handicapped. PMID- 2969886 TI - Setting up and running a disabled living centre. PMID- 2969887 TI - Disabled people and public transport. PMID- 2969888 TI - Insulin receptor signaling. Activation of multiple serine kinases. PMID- 2969889 TI - Modulation of interleukin-1 receptor expression and interleukin-1 response in fibroblasts by platelet-derived growth factor. AB - Our studies have revealed that the interleukin-1 (IL-1) receptors and response in BALB/c/3T3 fibroblasts are modulated by platelet-derived growth factor (PDGF). Incubation of quiescent cultures of BALB/c/3T3 fibroblasts with PDGF resulted in 4-5 fold higher 125I-IL-1 binding than the untreated cultures. Scatchard analysis showed that the increased 125I-IL-1 binding by PDGF-treated cells was due to a net increase in cell surface IL-1 receptors with no apparent change in binding affinity. The PDGF-induced increase of 125I-IL-1 binding was blocked by inhibitors of transcription, suggesting that a transcriptional event, perhaps a de novo synthesis of IL-1 receptors, is required for PDGF response. In a culture medium depleted of serum factors, IL-1 alone produced only a marginal (40-50%) increase in DNA synthesis in BALB/c/3T3 cells. In cells first exposed to PDGF, IL 1 produced 8-10-fold higher DNA synthesis than the controls. These findings provide evidence that IL-1 action in fibroblast may be regulated at the level of receptor expression. PMID- 2969890 TI - Growth regulation by GTP. Regulation of nucleotide pools in Neurospora by nitrogen and sulfur control systems. AB - Purine nucleotide pools in the fungus Neurospora crassa decline in response to carbon, nitrogen, or sulfur deprivation. There is, in addition, a decline in GTP/ATP ratios on nitrogen or sulfur deprivation in wild type. The GTP/ATP decline is missing on nitrogen deprivation of the nitrogen control mutant, nit-2, and on sulfur deprivation of the sulfur control mutant, cys-3. The nit-2 mutant also shows elevated UTP pools on nitrogen deprivation when compared with similarly treated wild type. Six-hour sulfur-deprived cys-3 shows multiple aberrations in nucleotide pools when compared with similarly treated wild type. These include very low energy charge and depletion of pools of most nucleotides. ATP in sulfur-deprived cys-3 drops by about 88%. Sulfur-deprived cys-3 is also greatly impaired in comparison with wild type in its ability to resume growth when restored to nutritional sufficiency after a period of sulfur deprivation. These results clearly demonstrate that the nitrogen (nit-2) and sulfur (cys-3 regulatory systems are not limited to control of catabolism of exogenous nitrogen and sulfur sources, respectively, but rather influence, a broader range of cellular properties than has been previously thought. The pattern of GTP pool control is consistent with a positive role for GTP in growth control. Evidence in other systems supporting such a growth-regulating role for GTP is discussed. PMID- 2969891 TI - One-chain urokinase-type plasminogen activator from human sarcoma cells is a proenzyme with little or no intrinsic activity. AB - We have compared the plasminogen activating capacity of one- and two-chain urokinase-type plasminogen activator (u-PA). In a 125I-plasminogen conversion assay in the presence of high amounts of a plasmin inhibitor, one-chain u-PA pretreated with diisopropyl fluorophosphate had no detectable activity, the detection limit corresponding to the activity of a 400-fold lower amount of two chain u-PA. In coupled assays in which generated plasmin was measured with a synthetic substrate, activity was clearly observed with the one-chain preparation, but the initial rate of plasminogen activation was lower than that of a 250-fold smaller concentration of two-chain u-PA. The coupled assays for one chain u-PA are self-activating because plasmin catalyzes conversion of one- to two-chain u-PA, and it is not possible to decide whether the low activity of one chain u-PA observed with this type of assay is intrinsic or due to contaminations. On the basis of these findings and a discussion of previous studies, it is concluded that one-chain u-PA has a variety of properties similar to the one-chain proenzyme forms of other serine proteases and that it should, therefore, be considered as a genuine proenzyme form of u-PA. PMID- 2969892 TI - Expression and characterization of a functional human insulin-like growth factor I receptor. AB - Stable transfectants of Chinese hamster ovary (CHO) cells were developed that expressed the protein encoded by a human insulin-like growth factor I (IGF-I) receptor cDNA. The transfected cells expressed approximately 25,000 high affinity receptors for IGF-I (apparent Kd of 1.5 X 10(-9) M), whereas the parental CHO cells expressed only 5,000 receptors per cell (apparent Kd of 1.3 X 10(-9) M). A monoclonal antibody specific for the human IGF-I receptor inhibited IGF-I binding to the expressed receptor and immunoprecipitated polypeptides of apparent Mr values approximately 135,000 and 95,000 from metabolically labeled lysates of the transfected cells but not control cells. The expressed receptor was also capable of binding IGF-II with high affinity (Kd approximately 3 nM) and weakly recognized insulin (with about 1% the potency of IGF-I). The human IGF-I receptor expressed in these cells was capable of IGF-I-stimulated autophosphorylation and phosphorylation of endogenous substrates in the intact cell. This receptor also mediated IGF-I-stimulated glucose uptake, glycogen synthesis, and DNA synthesis. The extent of these responses was comparable to the stimulation by insulin of the same biological responses in CHO cells expressing the human insulin receptor. These results indicate that the isolated cDNA encodes a functional IGF-I receptor and that there are no inherent differences in the abilities of the insulin and IGF-I receptors to mediate rapid and long term biological responses when expressed in the same cell type. The high affinity of this receptor for IGF-II also suggests that it may be important in mediating biological responses to IGF II as well as IGF-I. PMID- 2969893 TI - Liver (B-type) phosphofructokinase mRNA. Cloning, structure, and expression. AB - Mouse liver mRNA enriched in sequences coding for liver phosphofructokinase by polysome immunoadsorption was used as a template for the synthesis of cDNA. The double-stranded cDNA was inserted into the expression vector lambda gt11 and cloned. Preliminary identification of clones containing cDNA sequences for phosphofructokinase was made by screening the library with anti-rat liver phosphofructokinase serum and horseradish peroxidase-conjugated goat anti-rabbit IgG as second antibody. Subsequently, by selecting antibodies specific to fusion proteins expressed by putative clones and by reacting with Western blots of mouse liver proteins several clones were positively identified as containing liver phosphofructokinase sequences. A cDNA clone corresponding to 2708 nucleotides of liver phosphofructokinase mRNA was further characterized and sequenced. The liver phosphofructokinase mRNA has an open reading frame of 2343 nucleotides followed by a 3'-untranslated region of 303 nucleotides. The G/C-rich (76%) portion of the 5'-untranslated region precedes a characteristic translational start site of CCGCC(AUG). The mRNA coding sequence indicates that the liver phosphofructokinase subunit is composed of 780 amino acid residues and has a Mr of 85,000. Comparison of the deduced amino acid sequence of mouse liver phosphofructokinase with the known rabbit muscle phosphofructokinase shows 68% homology. The N-half of the liver phosphofructokinase has conserved substrate binding sites for ATP and fructose-6-P. The 25 C-terminal residues, which contain the ATP inhibitory site, are the least homologous (20%) but contain a putative phosphorylation site (Arg Arg-X-X-Ser). The liver phosphofructokinase mRNA is under nutritional and hormonal regulation. The liver phosphofructokinase mRNA level increased 4-fold when previously starved mice were refed a high carbohydrate, fat-free diet. This increase in mRNA level was blocked by 50% by the administration of dibutyryl cAMP. The induction of liver phosphofructokinase mRNA by fasting/refeeding was also diminished in streptozotocin diabetic mice. PMID- 2969894 TI - Calcium dependence during single-cycle catalysis of the sarcoplasmic reticulum ATPase. AB - We have investigated the kinetic and thermodynamic properties of the Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum under conditions that result in a single transport cycle. Simultaneous addition of ATP and EGTA to sarcoplasmic reticulum vesicles, preincubated with calcium, resulted in a transient of intermediate species. In the presence of saturating Ca2+ levels, total E-P species reached a maximum of 2.3 nmol/mg at 100 ms, followed by a monoexponential decay with kobs = 3.6 s-1. The data are interpreted in terms of Ca2+ sequestration, either by occlusion as Ca2+ in the phosphorylated enzyme or chelation by EGTA. Maximum Ca2+ uptake was 8.3 nmol/mg with the release of 4.4 nmol/mg Pi. The ratio of Ca2+ uptake to Pi release approached 1.9 over a wide [Ca2+] range. Equilibrium Ca2+ binding, in the absence of ATP, showed a K0.5 of 0.88 microM with a Hill coefficient of 1.9. The Ca2+ concentration dependence of Ca2+ uptake during single-cycle catalysis showed a 10-fold enhanced affinity (K0.5 = 0.06 microM) and was noncooperative (nH = 0.9). Quench with excess EGTA (greater than 2 mM) decreased Ca2+ uptake to 1 nmol/mg, indicating an "off" rate of Ca2+ from high affinity sites that exceeds 100 s-1. The ATP concentration dependence for a single-cycle catalysis showed an apparent K0.5 of 1.1 microM, similar to that for ATP equilibrium binding. It is proposed that enzyme phosphorylation proceeds only following binding of a second calcium ion to externally oriented sites whose intrinsic affinity is in the same range as the calcium dependence of a single-cycle turnover. PMID- 2969895 TI - Murine interleukin 1 receptor. Direct identification by ligand blotting and purification to homogeneity of an interleukin 1-binding glycoprotein. AB - Functional receptors (IL1-R) for the proinflammatory cytokine interleukin 1 (IL1) were solubilized from plasma membranes of the NOB-1 subclone of murine EL4 6.1 thymoma cells using the zwitterionic detergent 3[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). Membrane extracts were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and "ligand blotted" with 125I-labeled recombinant human IL1 alpha in order to reveal proteins capable of specifically binding IL1. A single polydisperse polypeptide of Mr approximately equal to 80,000 was identified in this way, which bound IL1 alpha and IL1 beta with the same affinity as the IL1-R on intact NOB-1 cells (approximately equal to 10(-10) M). The IL1-binding polypeptide was only seen in membranes from IL1-R-bearing cells and did not react with interleukin 2, tumor necrosis factor alpha, or interferon. IL1-R was purified to apparent homogeneity from solubilized NOB-1 membranes by affinity chromatography on wheat germ agglutinin-Sepharose and IL1 alpha-Sepharose. Gel electrophoresis and silver staining of purified preparations revealed a single protein of Mr approximately equal to 80,000 which reacted positively in the ligand-blotting procedure and which we identify as the ligand binding moiety of the murine IL1-R. Purified IL1-R exhibited the same affinity and specificity as the receptor on intact cells. The relationship of this protein to proteins identified by covalent cross-linking studies is discussed. PMID- 2969896 TI - A discontinuous factor H binding site in the third component of complement as delineated by synthetic peptides. AB - Factor H, a very important regulator of alternative pathway activation, exerts its effects by binding to the third component complement, C3. In this study we present evidence that factor H reacts with at least two sites in the third component of complement (C3), and we have mapped one of these sites within the C3d fragment of C3. By using direct binding assays of an anti-human H anti idiotypic antibody (alpha alpha H) and of H to C3 fragments, it was shown that both bound to the C3b and C3d (but not to C3c) fragments of C3. Cleavage of C3d by CNBr generated two major fragments with Mr values of 12,500 (residues 997 1107) and 8,600 (residues 1178-1252). Binding studies with these two fragments showed that only the Mr 8,600 fragment bound to both H and alpha alpha H. Several synthetic peptides (A58, 1192-1249; P28, 1187-1214; P16, 1194-1209; P14, 1201 1214; B17, 1206-1222; J28, 1222-1249; and J16, 1234-1249) were synthesized according to the primary sequence of the Mr 8,600 fragment. Based on the differential binding of these synthetic peptides to H, their inhibitory effect on H binding to C3b or C3d, and their effect on H cofactor activity, we mapped the H binding site in C3 to a discontinuous site spanning residues 1187-1249 of the C3 sequence. By studying the inhibition of H binding to C3b or C3d by the different synthetic peptides, we also present evidence that a second binding site in C3b for H exists. PMID- 2969897 TI - Serotonin and beta-thromboglobulin release reaction from platelet as triggered by interaction with polypeptide derivatives. AB - The effect of wettability of 14 polypeptide derivatives upon adhesion and activation of platelets was investigated with reference to release reactions from adhered platelets, using radioisotope labeling and radioimmunoassay method. The serotonin release was more significant from platelets adhered to polymer materials to which a large number of platelets are adhered. However, no clear relationship was found between adhesion of platelets and beta-thromboglobulin release from adhered platelets. Therefore, stimuli inducing serotonin release and beta-thromboglobulin release were considered to be from different origins. The trend in beta-thromboglobulin release was well correlated with the extent of morphological change of adhered platelets as observed by scanning electron microscope. It was suggested that the determination of released beta thromboglobulin in association with the measurement of platelet adhesion could be useful for evaluation of blood compatibility of materials. PMID- 2969898 TI - Preventing diabetic nephropathy in 1987: what to do until the data come. PMID- 2969899 TI - Diabetic nephropathy: is dietary protein harmful? AB - The suggested harmful effect of dietary protein on renal function in diabetic nephropathy was tested in three groups of insulin-dependent diabetic patients: 1) 10 patients without signs of nephropathy in spite of at least 30 years of diabetes; 2) 11 patients with nephropathy and reduced but stable glomerular filtration rate (GFR) (decline less than 4 ml/min per year [mean 1.8] during the last 2 years); 3) 10 patients with progressive nephropathy with GFR declining by an average of 11 ml/min per year. Dietary protein intake was estimated from a dietary history interview, as well as from urinary excretion of nitrogen (mean = 4.7 samples). Both methods showed a wide range of protein intake in all three groups of patients (0.6-2.3 g/kg body weight [BW]). The mean values did not differ between the groups, 1.30, 1.34, and 1.24 g/kg BW by interview, and 1.20, 1.10, and 1.13 g based on urinary nitrogen levels. There was no correlation between rate of decline of GFR and protein intake, even in those patients with no or minimal decline. These results do not support the hypothesis that dietary protein is a factor of importance in the development or progression of human diabetic nephropathy. PMID- 2969900 TI - Correlation of renal histopathology duration of diabetes, and control of blood glucose in patients with type II diabetes. AB - Correlations of renal histopathologic changes, duration of diabetes, and the degree of control of blood glucose in patients with diabetic nephropathy were determined. Biopsies from 17 patients with diabetic nephropathy due to Type II diabetes were examined. Mild histopathologic changes in renal tissues were observed in patients with good blood glucose control, even if their disease was of more than 5 years' duration. Moderate to severe renal histopathology was found in patients with poor blood glucose control regardless of the duration of diabetes. It is concluded that strict control of blood glucose during the course of diabetes is important in inhibiting the progression of renal involvement in patients with Type II diabetes. PMID- 2969901 TI - Antihypertensive treatment: long-term reversal of progression of albuminuria in incipient diabetic nephropathy. A longitudinal study of renal function. AB - This study was undertaken to clarify whether antihypertensive treatment has any effect on the rate of progression of kidney disease in patients with incipient diabetic nephropathy. Six insulin-dependent diabetic men with incipient nephropathy (urinary albumin excretion above 15 micrograms/min and total protein excretion below 0.5 g/24 h) were first given metoprolol (200 mg daily) with the subsequent addition of hydroflumethiazide. At the start of antihypertensive treatment, mean patient age was 32 +/- 4.2 years (SD) and mean duration of diabetes was 18 +/- 1.2 years. The patients were followed with repeated measurements of urinary albumin excretion for a mean of 5.4 +/- 3.1 years prior to, and for 4.7 +/- 1.3 years (SD) during treatment. Mean arterial blood pressure declined significantly during treatment, e.g., the values at 6 months before initiation of treatment being compared with values during the last 6 months of treatment fell from 107 mmHg +/- 7.6 to 93 +/- 3.8 (2p = 1.5%). Albumin excretion decreased from 131.0 micrograms/min X/divided by 2.9 (geometric mean X/divided by tolerance factor) to 41.7 micrograms/min X/divided by 2.9 (2p = 1.2%). Albumin clearance in per cent of glomerular filtration rate decreased from a mean of 0.0030 +/- 0.0019% (SD) to 0.0011 +/- 0.0010% (2p = 4.6%). The mean yearly increase in urinary albumin excretion before treatment was 18.0 +/- 17.0% (mean +/- SD); during treatment urinary albumin excretion decreased 19 +/- 10% per year (2p = 0.7%). No changes were seen in renal plasma flow (516 +/- 31.0 ml/min to 520 +/- 66 ml/min (n = 5)).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969902 TI - The progression of diabetic nephropathy in type I diabetics: relationship to metabolic control and blood pressure. AB - This study was designed to investigate the importance of risk factors such as hyperglycemia and elevated systolic and diastolic blood pressures on the progression of renal insufficiency in diabetics suffering from diabetic nephropathy. Seventeen patients with Type I, insulin-dependent diabetes mellitus (IDDM) (8 women and 9 men) undergoing chronic hemodialysis were investigated by retrospective follow-up and compared with 17 age and sex matched IDDM patients without diabetic nephropathy (controls). According to the time interval of creatinine increase from 200 to 600 mumol/l, the patients were divided arbitrarily into two groups with rapidly (group I less than 20 months) or slowly progressive (group II greater than or equal to 20 months) renal insufficiency. This period was 13.4 +/- 2.05 months in group I (age 36.67 +/- 2.47 years, diabetes duration 23.55 +/- 2.37 years) and 32.75 +/- 4.34 months in group II (age 40.62 +/- 2.63 years, diabetes duration 26.62 +/- 2.63 years, P.n.s.), respectively. The IDDM patients studied exhibited individually differing progressions of renal insufficiency at different times after manifestation of diabetes. After 15 years of diabetes duration, both risk factors, that is blood pressure and blood glucose concentrations, were elevated in nephropathic diabetics when compared with controls (p less than 0.01). During the phase of declining kidney function, mean blood pressures were found to be higher in IDDM patients with rapid progression of renal insufficiency when compared with slowly progressing diabetics. Although both risk factors were related to diabetic nephropathy, during the phase of renal insufficiency hypertension appeared to be more closely related to the further deterioration of kidney function. PMID- 2969903 TI - A solid phase enzyme immunoassay for the measurement of urinary albumin and the detection of microalbuminuria. AB - A test for the measurement of trace urinary albumin concentrations, which is suitable for the detection of microalbuminuria, was developed. The technique is an indirect enzyme-linked assay (ELISA) in which a fixed amount of anti-albumin antibody is placed into polystyrene tubes coated with human albumin, together with the urine sample to be tested. The albumin in the test specimen competes with the solid-phase albumin for binding to the added antibody. The test is precise (inter- and intra-assay coefficients of variation were 8.2% and 7.8%, respectively), accurate (mean recovery 102-106% for two human albumin preparations), and sensitive (detection limit 0.9 micrograms/ml). These characteristics are not dissimilar from those of the radioimmunoassay reported in the literature, with the advantages of being completely safe, easy to perform, and not requiring expensive equipment. Using this assay the urinary albumin excretion in 20 normal subjects was found to be 2.5 +/- 2.2 micrograms/min (range 0.9-7.5 micrograms/min) after 8 hours of bed rest and 4.5 +/- 5.7 micrograms/min (range 1.5-2.0 micrograms/min) after 8 hours of moderate physical activity. PMID- 2969904 TI - Double immunofluorescence studies of glomerular sialic acids in patients with diabetic nephropathy. AB - A study of double immunofluorescence staining of sialic acids and IgG in the glomeruli from patients with diabetic nephropathy is described. Renal biopsy specimens from seven patients with diabetic nephropathy were stained with rhodamine-labeled tricum vulgaris (WGA), limulus polyphemus (LPA), and FITC labeled anti-human IgG antiserum. Both binding of WGA and deposition of IgG in glomerular capillary walls were observed in all patients with diabetic nephropathy. However, the binding of LPA in tissue from the same patients was minimal, although deposition of IgG was marked. It is suggested that an accumulation of substances with an affinity for WGA (most probably N-acetyl glucosamine or N-acetyl neuraminic acid) occurs in the glomerular capillary walls of patients with diabetic nephropathy. PMID- 2969905 TI - Diabetic complications: patient experience and physician response. PMID- 2969906 TI - Near normoglycemia for 1 year has no effect on platelet reactivity, factor VIII, and von Willebrand factor in insulin-dependent diabetes mellitus: a controlled trial. AB - The impact of prolonged near-normoglycemia on platelet reactivity (spontaneous and induced platelet aggregation), factor VIII, and von Willebrand factor in patients with insulin-dependent diabetes mellitus (IDDM) was evaluated in a prospective, randomized, controlled clinical trial. Twenty IDDM patients with no or only minor clinical signs of microvascular disease were randomly assigned to 1 year of continuous subcutaneous insulin infusion (CSII) or unchanged conventional insulin treatment (CIT). Hemoglobin A1c declined during the 12 month observation period from 7.3 +/- 1.2% to 6.4 +/- 0.9% (2p less than 0.01) in the CSII group, while this measure of glycemic control was unchanged in the CIT group: 7.2 +/- 1.1% vs 8.0 +/- 1.6% (NS). Platelet reactivity, factor VIII, and von Willebrand factor concentrations were identical in the two groups at entry into the study, and no significant changes in these variables were seen in either group. Thus, the present results do not support the concept of increased platelet reactivity following CSII treatment. PMID- 2969907 TI - Effect of unilateral nephrectomy on three patients with histopathological evidence of diabetic glomerulosclerosis in the resected kidney. AB - Diabetes mellitus and surgical ablation of renal tissue are two independent influences associated with hyperfiltration and elevated levels of the glomerular transcapillary hydraulic pressure differential (delta P). There is increasing evidence that hyperfiltration with elevated delta P is pathogenic and leads to glomerular damage. The authors questioned whether these two influences (surgical ablation of renal tissue and diabetes mellitus) would act in an additive fashion in human patients to produce an accelerated decline in renal function. Three patients with non-insulin-dependent diabetes mellitus who had undergone a unilateral nephrectomy (for a variety of reasons) were (retrospectively) identified. In each patient, morphologic evidence of diabetic glomerulonephropathy was present in the resected kidney. The charts from these patients were reviewed and post-nephrectomy renal function was estimated over time by plotting reciprocal serum creatine values versus time. Follow-up intervals after nephrectomy varied from 4 to 15 years. The results of our follow up showed no obvious detrimental effect on renal function (as measured by 1/serum creatinine) attributable to the unilateral nephrectomy. The authors conclude that residual renal function (and ultimate outcome) in patients with non-insulin dependent diabetes mellitus is highly variable, but does not seem to be adversely affected (at least over the time span of observation in these patients) by unilateral nephrectomy. PMID- 2969908 TI - Assessing co-morbidity in the uremic diabetic. PMID- 2969909 TI - Microalbuminuria in insulin-dependent diabetes: prevalence and practical consequences. AB - Urinary albumin excretion in a representative sample of 679 patients with Type I (insulin-dependent) diabetes, 18 to 50 years of age, was investigated. Patients on antihypertensive therapy were excluded. Urinary albumin excretion was examined in one 24 hour urine sample using an ELISA technique. Twenty-three per cent of the patients had microalbuminuria, i.e., 30-300 mg albumin/24 h. The prevalence of microalbuminuria was independent of sex, age, insulin dose and diabetes duration. In the majority of those cases in which microalbuminuria was found during the first 10 years of diabetes, the concentrations were in the lower range, i.e., 30-50 mg/24 h. The prevalence of incipient nephropathy (urinary albumin excretion in a single urine sample of 51-300 mg/24 h) increased with diabetes duration. In patients with incipient nephropathy hemoglobin A1c tended to be, and blood pressure was, elevated compared with age, sex, and duration matched patients with normal urinary albumin excretion rates (p = 0.08 and p less than 0.001, respectively). Urinary albumin excretion and blood pressure were significantly correlated in the total group (n = 401, r = 0.2, p less than 0.001). On the basis of these findings practical guidelines for the handling of patients with microalbuminuria are proposed. PMID- 2969911 TI - Diabetic nephropathy: a clinical study of 498 patients. AB - The clinical pattern of nephropathy was studied in 498 diabetic patients who were hospitalized during the period 1980-1985 at the Christian Medical College Hospital, Vellore, India. The diagnosis of nephropathy was confirmed in the presence of persistent proteinuria of 500 mg or more in any 24 hour urine sample in the absence of urinary infection and congestive failure. Only four patients had Type I diabetes, all the rest being non-insulin-dependent (Type II). There was significant male preponderance, and the largest number of patients were in the fifth and sixth decades. The age of onset in Type II diabetics was between 30 to 50 years in 74%; 55% of the patients were known to have diabetes for less than 10 years, and in only 6% was the duration greater than 20 years. The degree of renal failure and proteinuria showed a variable pattern in relation to the duration of diabetes. Arterial hypertension was present in 80% of the patients and coronary artery disease in 33.5%, while cerebrovascular disease and peripheral vascular disease were evident in 7.4 and 4.8%, respectively. Fundoscopic examination showed evidence of retinopathy in 278 patients (53%), with proliferative changes in 17%. Clinical evidence of retinopathy was absent in 110 patients (22%), and in the rest the results of fundus examination was not documented; thus, the incidence of clinical retinopathy in this review was 72% (278/388). It is concluded that contrary to what has been observed in Type I diabetes, the progression of nephropathy in Type II bears no relationship to the duration of disease, nor is retinopathy a constant feature. PMID- 2969910 TI - Metabolic control in newly kidney transplanted insulin-dependent diabetics: improvement by insulin pump treatment (CSII). AB - Management of glucose homeostasis in newly kidney transplanted insulin-dependent diabetic patients is difficult. To examine whether continuous subcutaneous insulin infusion (CSII) could reverse this problem, six consecutive kidney transplanted Type I diabetic patients either proceeded with conventional insulin therapy (CIT) or were changed to CSII beginning on the third postoperative day. After a mean of 13 days, the insulin administration mode was changed from CIT to CSII (n = 3) or from CSII to CIT (n = 3), and continued for a further 15 days. Mean blood glucose calculated on the basis of four daily measurements (8.00, 12.00, 17.00, 22.00 h) during the study periods was significantly lower during CSII (8.0 +/- 0.4 mmol/l, mean +/- SEM) than on CIT (11.0 +/- 0.6 mmol/l; p less than 0.005). Moreover, the variability of blood glucose expressed as the M-value was lower during the pump treatment compared to CIT (p less than 0.001), while the number of blood glucose values below 3.0 mmol/l was similar (3.8 vs. 4.4%). Diurnal metabolic and hormonal profiles were twice determined on each regimen with 2 hourly sampling. Glycemic control was again found to be improved during CSII therapy as compared to CIT (p less than 0.01 or 0.05 less than p less than 0.10). Moreover, insulin pump treatment resulted in a significant reduction of two major intermediary metabolites, lactate and glycerol (p less than 0.05 and p less than 0.01, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969912 TI - Effect of continuous subcutaneous insulin infusion on kidney function and size in IDDM patients: a 2 year controlled study. AB - Glomerular filtration rate (GFR), renal plasma flow (RPF), kidney volume, and urinary albumin excretion rate were measured in 24 insulin-dependent diabetics, aged 29 +/- 7 years (mean +/- SD) with diabetes duration of 8 +/- 4 years who were randomly allocated to either continuous subcutaneous insulin infusion (CSII) (n = 12) or unchanged conventional insulin treatment (CIT) (n = 12). GFR, RPF, and kidney volume were identical but significantly increased above normal values in the two groups at the start of the study. After 24 months of CSII treatment, significant reduction in GFR was seen compared to pretreatment values (145 +/- 21 ml/min vs 139 +/- 21 ml/min, 2p less than 5.0%). However, RPF was not reduced after 24 months of CSII treatment (608 +/- 104 ml/min vs 601 +/- 106 ml/min). In the CIT group no changes in GFR or RPF was seen, and kidney volume remained unchanged in both groups; urinary albumin excretion was normal or near normal in both groups and remained unchanged. Thus, improved glycemic control in long-term IDDM patients is associated with normalization (reduction) of renal "hyperfunction," but despite this, no reduction was seen in the associated nephromegaly. PMID- 2969913 TI - Incidence of retinopathy in type I (insulin-dependent) diabetes: association with clinical nephropathy. AB - The association between the incidence of diabetic retinopathy and the development of diabetic nephropathy was studied in 110 Type I (insulin-dependent) diabetic patients during a period from 10 years before to 5 years after the onset of persistent proteinuria. This group of patients was compared with 110 diabetic patients, who were matched according to sex, age, and diabetes duration, but who were without proteinuria during the observation period. The cumulative incidence of proliferative retinopathy was 74% in patients with clinical nephropathy and 14% in patients free of proteinuria. The incidence of background retinopathy was 93% and 37%, respectively, and the incidence of retinopathy increased dramatically 5 years before the onset of proteinuria. Neither gender, age at onset of diabetes, nor blood pressure seemed to have much influence on the incidence of severe retinopathy. It is concluded that development of clinical diabetic nephropathy implies an extremely high risk of developing severe retinopathy. A common pathogenetic link may be suspected. PMID- 2969914 TI - Complement, C receptors, and immune complex disease. PMID- 2969916 TI - [Food restriction during the growth stage is related to the extension of life span]. PMID- 2969915 TI - Rapid separation of free fatty acids from neutral lipids in brain extracts. PMID- 2969917 TI - Lexical and semantic priming deficits in patients with Alzheimer's disease. AB - Two experiments utilizing priming procedures examined the status of semantic memory in demented and amnesic patients. In the first investigation, lexical priming was assessed in patients with dementia of the Alzheimer type (DAT), Huntington's Disease (HD), alcoholic Korsakoff's syndrome (KS), and in intact control subjects. Subjects were first exposed to a list of words in a rating task and then required to complete three-letter stems with the "first word that comes to mind". Half of the stems could be completed with the previously presented words and the other half were used to assess baseline guessing rates. Recall and recognition of incidentally exposed words was also assessed. Although all three patient groups were impaired on tests of recall and recognition, only the DAT patients exhibited a priming deficit on the stem-completion task. In the second experiment, DAT, HD, and intact control subjects were administered a semantic priming test which required the subject to "free associate" to the first words of previously presented semantically associated word pairs. The results for this association task showed that DAT patients were significantly less likely to produce the second word of the semantically related pair than were the other subject groups. The results of these two experiments suggest that the memory capacities of DAT patients are characterized by a breakdown in the structure of semantic memory and that this impairment is evident on some "automatic" as well as "effortful" processing tasks. PMID- 2969918 TI - Identification of a high-affinity receptor for interleukin 1 alpha and interleukin 1 beta on cultured human rheumatoid synovial cells. AB - In this report the binding of recombinant human interleukins 1 alpha and 1 beta (rIL-1 alpha and rIL-1 beta) to primary cultures of human rheumatoid synovial cells is measured and compared to the concentrations of these mediators required for stimulation of PGE2 production by these same cells. The average concentration of IL-1 alpha required for half-maximal stimulation of PGE2 was 4.6 +/- 1.5 pM (+/- SEM) (n = 6), whereas for IL-1 beta half-maximal stimulation was observed at a concentration of 1.3 +/- 0.24 pM (n = 6). Both direct and competitive binding experiments were performed. In direct binding experiments, IL-1 alpha bound with a Kd of 66 pM (n = 1), while IL-1 beta bound with a Kd of 4 pM (n = 2). In competitive binding experiments, IL-1 alpha inhibited binding of 125I-IL-1 alpha with a Ki of 33-36 pM (n = 2) and binding of 125I-IL-1 beta with a Ki of 51-63 pM (n = 2). IL-1 beta inhibited binding of 125I-IL-1 alpha with a Ki of 2-3 pM (n = 2) and binding of 125I-IL-1 beta with a Ki of 7 pM (n = 2). The binding data were best fit by a model specifying a single class of receptors with homogeneous affinity for either IL-1 alpha or IL-1 beta and with an abundance of 3,000-14,000 sites per cell. Autoradiography showed that the vast majority of the synoviocytes within the cultures possessed IL-1 receptors. Comparison of biological response curves with the binding curves indicates that the observed receptors exhibit sufficiently high affinity to mediate the response of human synoviocytes to low picomolar concentrations of IL-1 alpha and IL-1 beta. PMID- 2969919 TI - Molecular cloning and characterization of human cardiac alpha- and beta-form myosin heavy chain complementary DNA clones. Regulation of expression during development and pressure overload in human atrium. AB - We have constructed and characterized two types of myosin heavy chain (MHC) cDNA clones (pHMHC2, pHMHC5) from a fetal human heart cDNA library. Comparison of the nucleotide and deduced amino acid sequences between pHMHC2 and pHMHC5 shows 95.1 and 96.2% homology, respectively. The carboxyl-terminal peptide and 3' untranslated (3'-UT) regions are highly divergent and specific for these cDNA clones. By using the synthetic oligonucleotide probes that are complementary to the unique 3'-UT regions of these cDNA clones, we demonstrate that pHMHC2 is exclusively transcribed in the atrium, whereas the mRNA for pHMHC5 is predominantly expressed in the ventricle. This result indicates that pHMHC2 and pHMHC5 code for alpha- and beta-form MHCs, respectively. Furthermore, we show that beta-form MHC mRNA is expressed in adult atrium at a low level but scarcely expressed in fetal atrium. Finally, we demonstrate that MHC isozymic transition in pressure-overloaded atrium is, at least in part, regulated at a pretranslational level. PMID- 2969920 TI - Defect in the membrane expression of high affinity 72-kD Fc gamma receptors on phagocytic cells in four healthy subjects. AB - Three different receptors for the Fc portion of IgG (FcR) have been characterized on human leukocytes. We have identified four healthy members of one family, whose blood phagocytic cells lack functional 72 kD high-affinity FcRI. Their monocytes were unable to bind the Fc portion of mouse (m)-IgG2a and of monomeric human IgG, and they were unreactive with two anti-FcRI monoclonal antibodies. Thus, FcRI is either absent, expressed at very low density, or is so structurally altered as to be unable to bind both its ligand and the anti-FcRI antibodies. The failure to bind the Fc portion of mIgG2a underlies the previously reported inability of these monocytes to support T cell mitogenesis on OKT3 stimulation. FcRI was not inducible upon incubation of their monocytes or neutrophils in gamma interferon. However, their monocytes were able to bind aggregated human IgG, and to phagocytose IgG-coated particles in vitro. Both functions could be blocked with a monoclonal antibody to the 40-kD low-affinity FcRII and therefore apparently were mediated exclusively through FcRII. This also demonstrates that FcRII can mediate phagocytosis independently. Despite the FcRI defect, these subjects had no circulating immune complexes, no evidence of autoimmune pathology and no increased susceptibility to infections. PMID- 2969922 TI - Reduction of atherosclerosis by administration of dehydroepiandrosterone. A study in the hypercholesterolemic New Zealand white rabbit with aortic intimal injury. AB - Dehydroepiandrosterone (DHEA) is an endogenous steroid that blocks carcinogenesis, retards aging, and exerts antiproliferative properties. In vitro, it is a potent inhibitor of glucose-6-phosphate dehydrogenase, the first committed step of the pentose phosphate pathway. In man, serum levels of DHEA and its sulfate peak in early adulthood and drop markedly with age. Epidemiologic evidence indicates that low levels of DHEA or its sulfate conjugate are linked to an increased risk of developing cancer or of death from cardiovascular disease. Like cancer, atherosclerosis is a proliferative process characterized by both initiation and promotion phases. This similarity provided a framework in which to study the antiatherogenic effects of DHEA. Rabbits were randomly assigned to four groups. Two groups of rabbits received aortic endothelial injury by balloon catheter and were fed a 2% cholesterol diet for 12 wk. DHEA, 0.5%, was incorporated into the diet of one group receiving the 2% cholesterol diet and endothelial injury and also into the diet of one of the control groups. Animals were killed after 12 wk and aortas, hearts, and livers were studied. Plasma samples were analyzed for total cholesterol, VLDL, LDL, HDL, triglycerides, DHEA, and DHEA-sulfate levels. The atherogenic insult resulted in severe atherosclerosis in animals not treated with DHEA. In those receiving DHEA there was an almost 50% reduction in plaque size (P = 0.006), inversely related to the serum level of DHEA attained. Fatty infiltration of the heart and liver were also markedly reduced. These beneficial actions were not attributable to differences in body weight gain, food intake, total plasma cholesterol or distribution of cholesterol among the VLDL, LDL, or HDL fractions. The results show that high levels of plasma DHEA inhibit the development of atherosclerosis and they provide an important experimental link to the epidemiologic studies correlating low DHEA sulfate plasma levels with an enhanced risk of cardiovascular mortality. PMID- 2969921 TI - Neutrophil and monocyte cell surface p150,95 has iC3b-receptor (CR4) activity resembling CR3. AB - Previous investigations of p150,95 (CD11c), the third member of the CD18 membrane glycoprotein family that includes CR3 (Mac-1 or CD11b) and LFA-1 (CD11a), had demonstrated that solubilized p150,95 bound to iC3b-agarose in a manner similar to isolated CR3. The current study showed that membrane surface p150,95 also expressed iC3b-receptor activity and was probably the same as the neutrophil receptor for iC3b- or C3dg-coated erythrocytes (EC3bi or EC3dg) that had been previously designated CR4. Normal neutrophil and macrophage CR4-dependent EC3bi rosettes were inhibited by monoclonal anti-p150,95, and cells from a patient with CD18 deficiency did not form CR4-dependent EC3bi rosettes. With neutrophils that bore large amounts of CR1 and CR3 and little p150,95, EC3bi were found primarily via CR1 and CR3, and demonstration of p150,95-dependent rosettes required large amounts of fixed iC3b, low-ionic strength buffer, and antibody blockade of CR1 and CR3. By contrast, culture-derived macrophages expressed eight times more p150,95 than did monocytes and EC3bi were bound to both p150,95 and CR3 when EC3bi bore small amounts of fixed iC3b and assays were carried out in isotonic buffer. Comparison of the amounts of CR1, CR3, and CR4 in various tissues by immunoperoxidase staining revealed that CR4 was the most abundant C3 receptor molecule on tissue macrophages, and suggested that CR4 might be involved in clearance of C3-opsonized particles or immune complexes. PMID- 2969923 TI - Severe edema associated with etretinate. PMID- 2969924 TI - Fish oil supplementation results in decreased hypertriglyceridemia in patients with psoriasis undergoing etretinate or acitretin therapy. AB - Although retinoid derivatives are an effective treatment for severe psoriasis, they result in systemic toxicity, including hyperlipidemia. In an attempt to reverse this retinoid-related hyperlipidemia in patients with psoriasis, a prospective 4-week pilot study of fish oil supplementation was carried out in 25 patients with psoriasis vulgaris receiving etretinate (Ro-10-9359) or acitretin (Ro 10-1670). Daily fish oil supplements containing 3 gm of omega-3 fatty acids (1.8 gm of eicosapentaenoic acid 20:5 omega 3, and 1.2 gm of docosahexaenoic acid 22:6 omega 3) were found to be effective in reducing hypertriglyceridemia, with a significant mean reduction from 215.6 +/- 92.5 to 156.9 +/- 58.5 mg/dl (-27%) when compared with controls (203.6 +/- 46.9 to 204.1 +/- 54.3 mg/dl). High density lipoprotein cholesterol levels increased from 41.4 +/- 10.5 to 46.1 +/- 10.8 mg/dl (+11%), and the ratio of total cholesterol to high-density lipoprotein cholesterol decreased from 6.6 +/- 1.9 to 5.9 +/- 1.7 (-11%). It is concluded that fish oil supplementation may prove a valuable adjunct to ameliorate the lipid changes secondary to retinoids. PMID- 2969925 TI - Restenosis after successful percutaneous transluminal coronary angioplasty: serial angiographic follow-up of 229 patients. AB - To further understand the temporal mode and mechanisms of coronary restenosis, 229 patients were studied by prospective angiographic follow-up on day 1 and at 1, 3 and 6 months and 1 year after successful percutaneous transluminal coronary angioplasty. Quantitative measurement of coronary stenosis was achieved by cinevideodensitometric analysis. Actuarial restenosis rate was 12.7% at 1 month, 43.0% at 3 months, 49.4% at 6 months and 52.5% at 1 year. In 219 patients followed up for greater than or equal to 3 months, mean stenosis diameter was 1.91 +/- 0.53 mm immediately after coronary angioplasty, 1.72 +/- 0.52 mm on day 1, 1.86 +/- 0.58 mm at 1 month and 1.43 +/- 0.67 mm at 3 months. In 149 patients followed up for greater than or equal to 6 months, mean stenosis diameter was 1.66 +/- 0.58 mm at 3 months and 1.66 +/- 0.62 mm at 6 months. In 73 patients followed up for 1 year, mean stenosis diameter was 1.65 +/- 0.56 mm at 6 months and 1.66 +/- 0.57 mm at 1 year. Thus, stenosis diameter decreased markedly between 1 month and 3 months after coronary angioplasty and reached a plateau thereafter. In conclusion, restenosis is most prevalent between 1 and 3 months and rarely occurs beyond 3 months after coronary angioplasty. PMID- 2969926 TI - Role of coronary collateral vessels during transient coronary occlusion during angioplasty assessed by hemodynamic, electrocardiographic and metabolic changes. AB - The clinical role of collateral vessels was evaluated during transient coronary occlusion by percutaneous transluminal coronary angioplasty in 22 patients with (8) and without (14) collateral vessels. Coronary occlusion pressure, the ratio of mean coronary occlusion pressure to mean aortic pressure and myocardial perfusion pressure at 40 s of balloon inflation were significantly higher in patients with than in patients without collateral vessels. The changes in left ventricular systolic and end-diastolic pressure, maximal rate of rise of left ventricular pressure (peak dP/dt) and maximal rate of fall of left ventricular pressure (negative peak dP/dt) during balloon inflation were less in patients with than in patients without collateral vessels. Myocardial lactate was produced in patients without collateral vessels but not in those with such vessels. Marked ST segment elevation in the electrocardiogram occurred in patients without collateral vessels but either ST segment depression or mild ST segment elevation was observed in patients with collateral vessels. This study indicates that collateral vessels limit myocardial ischemia during coronary occlusion, probably as a result of increased myocardial perfusion pressure. PMID- 2969927 TI - Diastolic function in patients with aortic stenosis: influence of left ventricular load reduction. AB - Pressure overload hypertrophy of the left ventricle due to aortic stenosis is associated with abnormalities of left ventricular isovolumic relaxation and early diastolic filling. The relative contribution of the hemodynamic load on the left ventricle to the impairment of diastolic function observed in this disorder remains poorly understood. To study this relation, the vasodilator nitroprusside was administered to eight patients with aortic stenosis and normal systolic function. The effect of a short-term reduction in left ventricular preload and afterload on left ventricular isovolumic relaxation and early diastolic filling was assessed by analysis of simultaneous micromanometer left ventricular pressure and radionuclide angiographic volume measurements. At baseline, left ventricular systolic and end-diastolic pressures were markedly elevated, and associated with prolongation of the time constant of left ventricular relaxation and depression of the left ventricular peak filling rate. Infusion of nitroprusside resulted in reduction of left ventricular systolic (204 +/- 31 to 176 +/- 31 mm Hg, p less than 0.05) and end-diastolic (31 +/- 8 to 18 +/- 6 mm Hg, p less than 0.05) pressures, with no associated improvement in time constant of left ventricular pressure decay (T) (68 +/- 25 to 80 +/- 37 ms, p = NS), T 1/2 (34 +/- 8 to 34 +/- 14 ms, p = NS), left ventricular peak filling rate (2.3 +/- 0.5 to 2.3 +/- 0.8 end-diastolic volume/s, p = NS) or time to left ventricular peak filling rate (150 +/- 50 to 144 +/- 37 ms, p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969929 TI - Efficacy of a new angioplasty catheter for severely narrowed coronary lesions. AB - Conventional over the wire dilation catheters may be unsuccessful in crossing coronary lesions that are severely narrowed. Hence, a new, extremely low profile coronary angioplasty catheter specifically designed to dilate such lesions was investigated. The catheter features a 2.0, 2.5 or 3.0 mm (inflated diameter) balloon mounted on a guide wire. The deflated profile of the 2.0 mm balloon measures 0.020 +/- 0.001 in. (0.51 +/- 0.03 mm). The catheter can be used in conjunction with 7F angiographic or 8F guide catheters. The catheter was used in 61 patients, aged 43 to 86 years, with predominantly Canadian Cardiovascular Society class III-IV angina. Dilation was attempted in 77 lesions. Lesion length averaged 5.7 +/- 3.1 mm (mean +/- 1 SD), minimal diameter 0.51 +/- 0.25 mm and internal vessel diameter 2.27 +/- 0.43 mm. Sixty lesions (78%) were successfully dilated to less than 50% residual stenosis with this catheter alone; nine lesions were further dilated with a larger balloon catheter. The new catheter was unable to cross 13 lesions (17%); only 2 of these lesions were subsequently crossed with a conventional over the wire system. On the other hand, the catheter was used after failure of conventional dilating catheters in 21 lesions and was successful in 16. The new catheter was particularly valuable for distal lesions and those demonstrating 90 to 99% diameter reduction. For all lesions crossed, stenosis decreased from 76 +/- 11 to 29 +/- 12% after 2.9 +/- 2.7 inflations and peak inflation pressure of 8.0 +/- 2.9 bar. Complications were rare; coronary occlusion occurred in two lesions (3%) and dissection in three lesions (4%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969928 TI - Responsiveness of plasma atrial natriuretic factor to short-term changes in left atrial hemodynamics after percutaneous balloon mitral valvuloplasty. AB - To assess the effect of short-term alteration of left atrial pressure and volume on the circulating plasma level of atrial natriuretic factor, 11 patients with left atrial hypertension due to mitral stenosis were studied at the time of percutaneous balloon mitral valvuloplasty. Hemodynamic measurements and plasma atrial natriuretic factor levels were obtained before, immediately (5 to 10 min) after and 24 h after valvuloplasty, and echocardiographic left atrial size was determined before and 24 h after valvuloplasty. Immediately after valvuloplasty, left atrial pressure decreased from 28 +/- 2 to 10 +/- 1 mm Hg (p less than 0.0005), mitral pressure gradient decreased from 20 +/- 2 to 7 +/- 1 mm Hg (p less than 0.0005), mitral valve area increased from 0.8 +/- 0.1 to 1.9 +/- 0.2 cm2 (p less than 0.0005) and plasma atrial natriuretic factor level rose from 249 +/- 42 to 348 +/- 50 pg/ml (p less than 0.01). This short-term rise in atrial natriuretic factor level may reflect a transient increase in left atrial pressure associated with balloon occlusion of the mitral valve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969930 TI - Endurance testing for evaluation of antianginal therapy with amlodipine, a calcium channel blocking agent. AB - The utility of using an endurance test as well as a maximal exercise test to assess the effect of amlodipine, a dihydropyridine calcium antagonist, was evaluated in 16 patients with angina pectoris. Amlodipine, 10 mg/day, was compared with placebo in a double blind crossover study. After a 2 week single blind placebo period, patients entered a double blind crossover phase alternating between 4 weeks of placebo and 4 weeks of amlodipine. The two 4 week periods were separated by a 1 week single blind placebo washout period. The efficacy of drug therapy was assessed by frequency of angina, nitroglycerin consumption, peak oxygen consumption during a maximal treadmill exercise test and endurance time during a separate exercise test performed at 70% of the peak work capacity determined before randomization. There was a reduction in angina frequency during the double blind placebo and amlodipine studies (single blind placebo 14 +/- 2 episodes/2 weeks, double blind placebo 7 +/- 2 episodes/2 weeks [p less than 0.005], amlodipine 6 +/- 3 episodes/2 weeks, [p less than 0.005]), whereas nitroglycerin consumption was reduced with amlodipine (single blind placebo 12 +/ 4 tablets/2 weeks, double blind placebo 8 +/- 3 tablets/2 weeks, amlodipine 5 +/ 3 tablets/2 weeks [p less than 0.01]). Amlodipine produced a significant increase in peak oxygen consumption (single blind placebo 18.7 +/- 1.1 ml/kg per min, double blind placebo 18.2 +/- 1.8 ml/kg per min, amlodipine 20.4 +/- 1.6 ml/kg per min [p less than 0.05]) and endurance time (single blind placebo 15.2 +/- 1.5 min, double blind placebo 15.8 +/- 2.1 min, amlodipine 20.2 +/- 2.5 min [p less than 0.005]).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969931 TI - Wheal-and-flare reactions induced by allergen and histamine: evaluation of blood flow with laser Doppler flowmetry. AB - Dermal blood flow was evaluated after skin prick test with histamine and allergen in six patients with seasonal allergic rhinitis. Blood flow was registered continuously for 60 minutes after the test procedure with laser doppler flowmetry, which allows noninvasive measurements. Blood flow was measured close to the skin test in the wheal obtained, and at a greater distance from the prick in the flare reaction. Tests were performed with preloaded skin test needles with histamine and the appropriate allergen freeze-dried on the point of the lancet, as well as with the appropriate negative control. The prick test procedure, by itself, induced a transient increase in blood flow that was normalized again after 9 minutes for the closest measurement. Histamine induced a rapid increase in blood flow in both the flare and wheal reaction that was normalized after about 45 minutes. The increase was significantly higher in the flare compared to the wheal for the time points from 6 1/2 to 13 minutes. Allergen induced a similar increase in blood flow. However, the increase was not noticeable until 2 1/2 minutes after the allergen application and was not completely abolished within 60 minutes. Furthermore, the difference between the flare and wheal reaction, with the higher values for the flare reaction, was present for a longer period of time than for the equivalent histamine measurements. In conclusion, laser doppler flowmetry appears useful for continuous evaluation of vascular changes induced at skin prick tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969932 TI - Laboratory evaluation of Bacillus sphaericus 1593-4 and preliminary field trials for control of Culex quinquefasciatus in septic tanks. PMID- 2969933 TI - Quantitative assessment of interleukin-2-producing alloreactive human T cells by limiting dilution analysis. AB - A limiting dilution (LD) culture system was developed to estimate the frequency of IL-2 producing alloantigen-reactive human helper T lymphocytes (HTL). E rosette-purified T cells or cell sorter-separated CD4+ and CD8+ subsets were co cultured under LD conditions with irradiated allogeneic Epstein-Barr virus transformed lymphoblastoid cell lines (LCL) in the absence of additional growth factors. IL-2 in microculture supernatants was detected in a bioassay combined with rapid colorimetric visualization (cleavage of MTT). Under these conditions, one out of 250-800 E+, 180-280 CD4+, and 440-1000 CD8+ T cells produced IL-2 after a culture period of 3 days. This quantitative analysis of alloantigen specific human HTL is complete within 4 days and thus provides a rapid method with potential applications in various clinical situations, e.g., transplantation medicine. PMID- 2969934 TI - Computerized data analysis in cellular immunology. Enhancement and suppression of immune responses. AB - Data obtained from assays for the evaluation of helper or suppressor activity are analyzed by statistical techniques that increase the accuracy and amount of information. The analysis is performed by a set of simple and ready to use computer programs which do not require statistical expertise and are available on request. PMID- 2969935 TI - [Myocardial and cerebral infarction in a 17-year-old man with hyper-lp (a) lipoproteinemia and hypercholesterolemia associated with nephrotic syndrome due to systemic lupus erythematosus]. PMID- 2969936 TI - [Distribution of muscle spindles in bat masticatory muscles and its significance]. PMID- 2969937 TI - Glutathione peroxidase activity, lipid peroxides and selenium status in blood in patients with Down's syndrome. AB - The concentrations of selenium and lipid peroxides and the catalytic activity of glutathione peroxidase were measured in the blood of 6 children (6-16 years of age) and 8 adults (17-27 years old) with Down's syndrome (trisomy 21). The values were compared with those for a control group of age-matched normal people. The selenium concentration in whole blood, erythrocytes and plasma was significantly lower in trisomy 21 patients than in normal subjects (p less than 0.001) in both age groups. No statistically significant differences were observed in selenium concentration in whole blood, erythrocytes and plasma between children and adults in the Down's syndrome group. Glutathione peroxidase catalytic activity in erythrocytes was significantly higher in Down's syndrome children than in healthy children (p less than 0.001). Plasma glutathione peroxidase catalytic activity in both investigated age groups was statistically considerably lower in the Down's syndrome patient group. The concentration of lipid peroxides, expressed as the malondialdehyde concentration, is lower in Down's syndrome patients. No correlation between selenium concentration, glutathione peroxidase catalytic activity and amount of lipid peroxides was found in the trisomy 21 patient group. PMID- 2969938 TI - Word identification in reading proceeds from spelling to sound to meaning. AB - Van Orden (1987) reported that false positive errors in a categorization task are elevated for homophonic foils (e.g., HARE for A PART OF THE HUMAN BODY). Two new experiments replicate this finding and extend it to nonword homophone foils (e.g., SUTE FOR AN ARTICLE OF CLOTHING). False positive errors to nonword homophone foils substantially exceed false positive errors to nonhomophonic nonword spelling controls, showing that the phonological characteristics of the nonword foils are critical. Because nonwords are not represented in the lexicon, this new result implicates computed phonological codes as a source of the categorization errors. Additionally, in each of two experiments, matched word and nonword homophones produced virtually identical error rates. If stimulus nonword homophones are viewed as extremely unfamiliar words, compared with the relatively familiar stimulus word homophones, then our failure to observe an effect of stimulus familiarity strengthens the case that phonological coding plays a role in the identification of all printed words. The fact that the results are obtained in a categorization task that requires reading for meaning (rather than a lexical decision task) makes it difficult to avoid the conclusion that phonological mediation plays a role in normal reading of text for meaning. PMID- 2969939 TI - Semantic, phonological, and mediated priming in reading and lexical decisions. AB - Four experiments were conducted to investigate whether semantic activation of a concept spreads to phonologically and graphemically related concepts. In lexical decision or self-paced reading tasks, subjects responded to pairs of words that were semantically related (e.g., light-lamp), that rhymed (e.g., lamp-damp), or that combined both of these relations through a mediating word (e.g., light damp). In one version of each task, test lists contained word-word pairs (e.g., light-lamp) as well as nonword-word (e.g., pown-table) and word-nonword pairs (e.g., month-poad); in another version, test lists contained only word-word pairs. The lexical decision and self-paced reading tasks were facilitated by semantic and rhyming relations regardless of the presence or absence of nonwords on the test lists. The effect of the mediated relation, however, depended on the presence of nonwords among the stimuli. When only words were included, there was no effect of the mediated relation, but when nonwords were included, lexical decision and self-paced reading responses were inhibited by the mediated relation. These inhibitory effects are attributed to processes occurring after lexical access, and the relative advantages of the self-paced reading task are discussed. PMID- 2969940 TI - Elaborative inferences during reading: do they occur on-line? AB - Four experiments were conducted to examine the extent to which readers construct elaborative inferences on-line during reading. In Experiment 1, gaze durations were measured while subjects read anaphors to target antecedents that referenced a particular category member either explicitly or implicitly. When the context strongly suggested a particular category member, gaze durations on an anaphor were the same following either an implicit or an explicit antecedent. When the context did not suggest any particular category member, gaze durations were significantly longer following an implicit antecedent. The results confirmed that, with sufficient context, readers will generate a simple elaborative inference on-line. These results were replicated in Experiment 2 in which the materials did not strongly signal the inference but a sentence designed to encourage subjects to infer was included. In Experiment 3, this "demand sentence" was not included, and readers did not appear to construct the targeted inference. The results of Experiment 4 confirmed that once generated, elaborative inferences are stored as part of the long-term-memory representation of a passage. PMID- 2969942 TI - Mental scanning in auditory imagery for songs. AB - Four experiments examined how people operate on memory representations of familiar songs. The tasks were similar to those used in studies of visual imagery. In one task, subjects saw a one-word lyric from a song and then saw a second lyric; then they had to say if the second lyric was from the same song as the first. In a second task, subjects mentally compared pitches of notes corresponding to song lyrics. In both tasks, reaction time increased as a function of the distance in beats between the two lyrics in the actual song, and in some conditions reaction time increased with the starting beat of the earlier lyric. Imagery instructions modified the main results somewhat in the first task, but not in the second, much harder task. The results suggest that song representations have temporal-like characteristics. PMID- 2969941 TI - Perceptual learning of synthetic speech produced by rule. AB - To examine the effects of stimulus structure and variability on perceptual learning, we compared transcription accuracy before and after training with synthetic speech produced by rule. Subjects were trained with either isolated words or fluent sentences of synthetic speech that were either novel stimuli or a fixed list of stimuli that was repeated. Subjects who were trained on the same stimuli every day improved as much as did the subjects who were given novel stimuli. In a second experiment, the size of the repeated stimulus set was reduced. Under these conditions, subjects trained with repeated stimuli did not generalize to novel stimuli as well as did subjects trained with novel stimuli. Our results suggest that perceptual learning depends on the degree to which the training stimuli characterize the underlying structure of the full stimulus set. Furthermore, we found that training with isolated words only increased the intelligibility of isolated words, although training with sentences increased the intelligibility of both isolated words and sentences. PMID- 2969943 TI - Picture memory: recognizing added and deleted details. AB - When people are presented simple and complex pictures and then tested in a same changed recognition test with a simple or complex form of each, d' is greater for the simple than the complex picture (Pezdek & Chen, 1982). The results of three experiments confirm the robustness of this "asymmetric confusability effect" and test a model of the processes underlying this effect. According to the model, pictures are schematically encoded such that the memory representation of both simple and complex pictures is similar to the simple form of each. In Experiment 1, a sentence was presented that described the central schema in the picture prior to subjects' viewing each picture. This manipulation exaggerated the asymmetric confusability effect; schematic processing thus underlies the effect. Results of Experiment 2 refute the hypothesis that the effect results from subjects erroneously anticipating a recall test rather than a recognition test. Furthermore, although some of the nonschematic elaborative information in complex pictures is stored in memory, it is difficult to retrieve to verify that something is missing when complex presentation pictures are changed to simple test pictures (Experiment 3). Thus, although people are able to distinguish large sets of old pictures from new distractor pictures, their ability to detect missing elaborative visual details is more limited. PMID- 2969944 TI - On the relation between perceptual priming and recognition memory. AB - Two experiments evaluated the hypothesis that perceptual fluency is used to infer prior occurrence. Subjects heard (Experiment 1) or saw (Experiment 2) a list of words and then were presented in the same modality with both these and other words twice in succession: first in a more or less impoverished fashion, and then in clear fashion. For the first of these two presentations, the subjects tried to identify the word; for the second, they gave a recognition judgement. As predicted by the perceptual fluency hypothesis, and as has been found in previous research, the recognition judgments were more positive for identified words than for unidentified words. However, degree of impoverishment, by which apparent perceptual fluency was brought under experimental control, did not affect the recognition judgments. The perceptual fluency hypothesis was therefore not supported, and the observed relation between identification and recognition was attributed to an item selection effect. PMID- 2969945 TI - Analogical transfer, problem similarity, and expertise. AB - When we encounter a new problem, we are often reminded of similar problems solved earlier and may use the solution procedure from an old problem to solve a new one. Such analogical transfer, however, has been difficult to demonstrate empirically, even within a single experimental session. This article proposes a framework for conceptualizing analogical problem solving that can account for the conflicting findings in the literature. In addition, the framework leads to two predictions concerning the transfer behavior of experts and novices. These predictions concern both positive and negative transfer and are based on the different types of features included in the problem representations of experts and novices. First, when two problems share structural features but not surface features, spontaneous positive transfer should be more likely in experts than in novices. Second, when two problems share surface but not structural features, spontaneous negative transfer should be stronger for novices than for experts. These predictions were supported by the results of three experiments involving college students solving a complex arithmetic word problem. PMID- 2969946 TI - The influence of expertise on X-ray image processing. AB - Observers with four different levels of radiological experience performed a recognition memory task on slides of faces and chest X-ray films. Half of the X ray films revealed clinically significant abnormalities and half did not. Recognition memory for faces was uniformly high across all levels of radiological experience. Memory for abnormal X-ray films increased with radiological experience and, for the most experienced radiologists, was equivalent to memory for faces. Surprisingly, recognition memory for normal films actually decreased with radiological experience from above chance to a chance level. These results indicate that radiological expertise is associated with selective processing of clinically relevant abnormalities in X-ray images. Expert radiologists appear to process X-ray images the way that we all process faces, by quickly detecting and devoting processing resources to features that distinguish one stimulus from another. However, the selective processing of X-ray films appears to be restricted to clinically relevant abnormalities. As they develop the ability to detect these abnormalities, radiologists appear to lose the ability to detect variations in normal features. PMID- 2969947 TI - A model of automatic attention attraction when mapping is partially consistent. AB - A model is described to account for the data of Durso, Cooke, Breen, and Schvaneveldt (1987). On the basis of the relative frequency of an item's presentation as a target, the item develops an automatic tendency to attract attention. When stimuli are then displayed, each calls the attention system to a degree determined by its present strength. We assume that attention eventually drifts to the strongest stimulus (which is then given as a response), but in a time determined inversely by the difference in strength between the two strongest stimuli. A version of this model in which the strengths were freely estimated parameters predicted the various elements of the data with good accuracy. In other versions of the model, strength values were derived from assumptions concerning the learning of automatism. Two of these models, quite different in character, captured the major qualitative features of the data. Further empirical tests of the models are suggested. PMID- 2969948 TI - Mechanisms by which the putative serotonin receptor antagonist metitepin alters nociception in mice. AB - The putative serotonin (5-HT) receptor antagonist metitepin (0.5 mg/kg, intraperitoneally) produced hypoalgesia in the increasing temperature hot-plate test and hyperalgesia in the tail-flick test in mice. The effects of metitepin were not altered after depletion of 5-HT by the neurotoxin 5,7 dihydroxytryptamine (5,7-DHT, 80 micrograms free base, intracerebroventricularly) or the serotonin synthesis inhibitor p-chlorophenylalanine (PCPA, 400 mg/kg for 10 consecutive days). After chronic administration (2 or 5 mg/kg for 18 consecutive days) tolerance to the effect of metitepin (0.5 mg/kg) and cross tolerance to the antinociceptive effect of the 5-HT agonist 5-methoxy-N,N dimethyltryptamine (5-MeODMT, 3 mg/kg) was found in the hot-plate test but not in the tail-flick test. It is suggested that metitepin may block descending 5-HT transmission while more complex mechanisms of action are involved at supraspinal level. One possibility is that metitepin exhibits partial agonist properties or, alternatively, that the drug may block 5-HT subsystems which tonically enhance nociception. PMID- 2969950 TI - Human dopamine receptor subtypes--in vitro binding analysis using 3H-SCH 23390 and 3H-raclopride. AB - Affinities and regional densities of the D1- and D2-dopamine receptor subtypes were studied in the human post-mortem brain in vitro using the two selective radioligands 3H-SCH 23390 and 3H-raclopride. 3H-Raclopride binding was confined to the caudate nucleus, the putamen and the substantia nigra, while 3H-SCH 23390 bound to cortical regions as well. The binding of 3H-SCH 23390 was reduced by a low concentration of ketanserin, indicating binding to 5-HT2 receptors in addition to the D1-dopamine receptors. The endogenous neurotransmitter dopamine interacted potently both with the D1-dopamine receptor and the D2-dopamine receptor, displaying two affinity states for each subtype. The distribution of the dopamine receptor subtypes obtained in the present in vitro investigation is in agreement with data obtained with 11C-SCH 23390 and 11C-raclopride in positron emission tomographic studies in human volunteers. PMID- 2969949 TI - Hypothermia induced by m-trifluoromethylphenylpiperazine or m chlorophenylpiperazine: an effect mediated by 5-HT1B receptors? AB - In the present study we examined the effect of different drugs on the m trifluoromethylphenylpiperazine (TFMPP)- and m-chlorophenylpiperazine (m-CPP) induced hypothermia in mice. Both the hypothermias studied are blocked or reversed by pindolol, cyanopindolol and compound 21-009, but not by atenolol. Neither hypothermia is antagonized by 5-HT1A antagonists (ipsapirone, spiperone), a 5-HT1C antagonist (mesulergine), 5-HT2 antagonists (cyproheptadine, mianserin, methysergide), 5-HT3 antagonists (ICS 205930, metoclopramide). The examined hypothermias are not antagonized by other antihypothermic agents (pimozide, idazoxan, atropine). The 8-OH-DPAT-induced hypothermia is not affected by cyanopindolol or compound 21009. The obtained results indicate that the TFMPP- and m-CPP-induced hypothermias in mice are mediated by 5-HT1B. These hypothermias may be a good screening test for evaluation of the 5-HT1B-agonistic and 5-HT1B antagonistic activity. PMID- 2969951 TI - Subcutaneous administration of lisuride in the treatment of complex motor fluctuations in Parkinson's disease. AB - 28 patients with Parkinson's disease showing complex "on-off" fluctuations in response to chronic levodopa plus dopa decarboxylase inhibitor (po) were treated with subcutaneous lisuride using a portable infusion pump. All patients improved initially during the first weeks of treatment. Four patients abandoned the trial within the first few weeks as a consequence of psychiatric complications (2 cases), inability to understand how to use the pump (one case) and subcutaneous nodule formation plus psychological rejections to wearing a pump (one case). All other 24 patients were treated for a minimum periods of 3 months (mean 9.6 months, maximum 24 months). The average daily dose of lisuride was 2.80 mg. The levodopa dose was reduced by 37%, but total withdrawal was not possible in any patient. Among the 18 patients who continued treatment at present, about 50% are independent and capable of undertaking most daily life activities. Psychiatric side-effects were present in 9 patients leading to permanent withdrawal in five. Subcutaneous lisuride infusions added to oral levodopa are clearly effective in patients with severe motor fluctuations. Careful selection of suitable patients and close monitoring is mandatory in order to obtain the best therapeutic results while reducing the risk of psychiatric adverse effects. PMID- 2969952 TI - Mapping dopamine receptors in the human brain. AB - We have investigated the anatomic localization of dopamine D1 and D2 receptors in the human brain using selective high affinity ligands for both types of dopamine receptors and the technique of receptor autoradiography. Dopamine D1 receptors were labeled in postmortem human brain tissue sections using the antagonist [3H]SCH 23390. Dopamine D2 receptors were labeled in consecutive tissue sections using the agonist [3H]205-502 and the antagonist [3H]spiroperidol. D1 and D2 dopamine receptors presented a heterogeneous distribution in the human brain. The highest concentrations of both D1 and D2 receptors were found in parts of the basal ganglia, particularly the nucleus caudatus and putamen. Lower concentrations were seen in other areas for example, the lateral globus pallidus was enriched in D2 receptors and the medial globus pallidus in D1 receptors. The substantia nigra contained intermediate densities of both D1 and D2, D1 receptors being present in higher concentrations. Dopamine D1 receptors were also localized in areas outside of the basal ganglia, particularly in the neocortex, amygdala and hippocampal formation. Dopamine D2 receptors were also present in areas outside of the basal ganglia, the most significant densities being found in the hippocampal formation. We observed a marked age-dependent decline in the density of D1 receptors during the first decades of life. In contrast, D2 receptor concentrations appeared to be unaltered with age. The distribution and densities of dopamine receptors were examined in 12 cases of Parkinson's disease and compared to a control adult population. No significant differences in density and distribution were seen for either D1 nor D2 receptors. PMID- 2969953 TI - Continuous subcutaneous lisuride infusions in Parkinson's disease. AB - Thirteen patients with idiopathic Parkinson's disease and "on-off" fluctuations on oral levodopa plus dopa decarboxylase inhibitor (DDI) were treated with continuous (24 hour) subcutaneous lisuride infusions together with a reduced dose of levodopa (plus DDI). An improvement in motor performance was seen in 10 patients, with a mean increase in percentage of waking time spent "on" of 32 per cent (range 13-59 percent). However, adverse effects were common, especially psychiatric effects, leading to treatment withdrawal in 11 of 13 subjects after a mean of 40 days' treatment. Continuous lisuride infusion together with a small dose of levodopa (plus DDI) are effective treatment for "on-off" fluctuations in Parkinson's disease, but the frequency of adverse effects limits the number of patients who can be treated successfully with this technique. PMID- 2969954 TI - Treatment of Parkinson's disease with subcutaneous lisuride infusions. AB - Four patients with Parkinson's disease and severe fluctuating responses to levodopa and oral dopamine agonists were treated with continuous administration of lisuride infusions, administered by means of an externally worn pump. Levodopa dosage ranged from 300 to 687 mg/day and was kept stable throughout the study. In addition increasing doses of lisuride were injected subcutaneously in the abdomen. Lisuride doses ranged from 41 to 104 micrograms/h. A marked improvement in mobility was observed in every patient while severe biphasic dyskinesais almost remitted in one of them. The most common side-effect was the presence of subcutaneous nodules appearing at the injection site. Two cases had mild hemorrhagic complications and one initially had nausea. One patient developed acute psychiatric disturbances severe enough to be excluded from the study. Our findings suggest that lisuride subcutaneous infusions can be useful in severily handicapped parkinsonian patients, however local and psychiatric side-effects may be a serious threat in the long-term care. PMID- 2969955 TI - Lisuride infusion pump in Parkinson's disease. A report of two cases. AB - Two patients, ages 66 and 72, with complications of chronic levodopa therapy (random fluctuations, end of dose deterioration and dyskinesias) who were treated with Lisuride by means of a portable subcutaneous infusion pump are reported. Results obtained show significant improvement in disability through a net increase in the number of hours spent "on". Dyskinesias remained unmodified. Limiting psychiatric side effects were observed in one of the patients. Practical and technical aspects of the management of this therapeutic method are discussed. PMID- 2969958 TI - Abdominal muscle paralysis from herpes zoster. PMID- 2969957 TI - The value of routine CSF analysis during lumbar radiculography for low back pain. AB - The medical notes of 370 orthopaedic patients with low back pain, who underwent lumbar radiculography, were retrospectively reviewed to determine the effect, if any, that CSF analysis had on their subsequent management. It was found that no apparent alteration of management occurred in a single patient and hence the routine sampling of CSF during lumbar radiculography for uncomplicated low back pain has been discontinued in our department. PMID- 2969956 TI - A prospective study of acute idiopathic neuropathy. III. Immunological studies. AB - The immune responses of 100 patients who presented with an acute idiopathic neuropathy were compared with those of age and sex matched controls. Blood lymphocytes and their subsets were counted with a fluorescent activated cell sorter. CD8+ (putative suppressor) lymphocytes were significantly reduced in the first week of the disease but total lymphocytes, total T and CD4+ (putative helper) cells were not altered. This reduction depended on the nature of the preceding infection. Serum complement C3 and C4 concentrations remained normal and immune complexes were rarely detected with a C1q binding assay. Complement fixing antibodies to human peripheral nerve antigens were discovered in the serum of 7% of patients but only 1% of controls. Complement-fixing antibodies to galactocerebroside were not discovered in any sera. Enzyme-linked immunoassays detected increased antibody responses to galactocerebroside but none at all to human P2 myelin protein in the patient sera. Forty microliter of serum from five patients injected into the sciatic nerves of rats did not induce significantly more demyelination than the serum from control patients. It is concluded that auto-immune responses can only be detected by these techniques in a small minority of patients with acute idiopathic neuropathy. PMID- 2969959 TI - Comparison of physiological and histochemical properties of motor units after cross-reinnervation of antagonistic muscles in the cat hindlimb. AB - 1. Physiological and histochemical properties of the cat ankle extensor muscles, the lateral and medial gastrocnemius, and the soleus were studied after cross reinnervation by flexor motoneurons. 2. Tibial and common peroneal nerves were cut and cross-united in the popliteal fossa of 2- to 6-mo-old cats. Eighteen to 24 mo later, single motor units were isolated by dissection and stimulation of ventral root filaments and classified into four types: fast-twitch, fatigable (FF), fast-twitch with intermediate fatigue resistance (FI), fast-twitch, fatigue resistant (FR), and slow, fatigue-resistant (S). Muscle fibers were classified as fast glycolytic (FG), fast, oxidative glycolytic (FOG), and slow oxidative (SO) on the basis of histochemical staining. 3. Although motor-unit force was normally well correlated with the size of the innervating motor axon in the cross reinnervated muscles, the force of different unit types overlapped considerably. The reinnervated motor units also showed a higher than normal degree of fatigability. 4. The range of muscle unit forces in cross-reinnervated triceps surae muscles was the same as in the normally innervated triceps surae muscles. This range is 2-3 times greater than the flexor muscles, which the common peroneal nerve normally supplies. The range of contraction speed of units in the cross-reinnervated extensor muscles was comparable to that in the flexor muscles, consistent with a motoneuron-specific determination of muscle speed (28). 5. SO and FOG muscle fibers were found in all reinnervated triceps surae muscles, but FG fibers were only found in reinnervated medial gastrocnemius (MG) and lateral gastrocnemius (LG) muscles, consistent with previous findings of the resistance of soleus muscles to complete conversion (10, 16, 20, 21). Type grouping of muscle fibers was characteristic of the reinnervated muscles. 6. Reinnervated SO muscle fibers were larger than the corresponding fibers in normally innervated muscles as were the estimated number of muscle fibers innervated by slow motor axons. Nonetheless, the force generated by the S motor units remained relatively smaller than FR and FF units. The relative contributions of the number, cross sectional area and specific tension to the force generation of reinnervated motor units are discussed. PMID- 2969960 TI - Fiber architecture and histochemistry in the cat neck muscle, biventer cervicis. AB - 1. Biventer cervicis (BC) is an anatomically complex muscle that is divided by tendinous inscriptions into five in-series compartments of motor units. We have analyzed the fiber architecture and fiber-type composition of these different compartments using microdissection and histochemical methods. 2. BC narrows as it runs rostrally, but its in-series compartments have similar cross-sectional areas. The tapered shape of BC comes about because tendinous inscriptions and the tendon of insertion are oriented obliquely and muscle fibers attach in a progressively offset fashion from the medial to the lateral muscle edge. 3. Individual compartments of BC differ from one another in their architecture. The rostral two compartments (1 and 2) contain fibers of similar length that run between two plates of tendinous tissue. Compartments 3 and 4 are divided into two or three in-parallel subvolumes whose fiber bundles differ in their lengths and sites of attachment. Compartment 5 is the most variable in its structure. In some cats it is separated from compartment 4 by a tendinous inscription, but in other cats, it blends with a dorsomedial part of compartment 4 to form a single subvolume. 4. The relative lengths of fibers in different compartments were analyzed when the head and neck were held in different postures. Fibers in rostromedial regions were stretched more effectively when the head was flexed at suboccipital joints, and appeared to be less sensitive to movements at lower cervical joints. Movements across lower cervical joints produced substantial length changes in caudolateral parts of BC. 5. Muscle fibers of different histochemical types were not distributed evenly within each muscle compartment. Slow, oxidative (SO) fibers accounted for the majority of fibers near the nuchal midline but for only 30%-45% of fibers in lateral muscle regions. Proportions of fast, glycolytic (FG) fibers were greatest in lateral regions. Fast, oxidative glycolytic (FOG) fibers were distributed quite uniformly throughout each compartment. 6. The specialized architecture of BC may shape its physiological capabilities. The complex internal structures of different compartments may alter the length-tension properties of BC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2969961 TI - For an elective surgical management in lumbar intervertebral disc herniation. Experience of 25 years of surgery. AB - On the base of a wide clinical experiences, the Authors report some consideration about relationship between clinical findings and neuroradiological pictures (plain film, myelography, CT and NMR) of the lumbar vertebral and disc diseases. It's pointed out the useful aspect of each imaging technique that contribute to obtain the more complete informations about anatomy of the lesion; such considerations allows the surgeon to plain the more adequate surgical technique. The Authors think myelography, still today, is the investigation of choice in this kind of pathology. PMID- 2969962 TI - Differential effects of adrenalectomy and starvation-refeeding on hepatic lipogenic responses to dehydroepiandrosterone and glucocorticoid in BHE and Sprague-Dawley rats. AB - The interaction of rat strain and glucocorticoid status on the dehydroepiandrosterone (DHEA)-mediated decrease in response to starvation refeeding was studied. DHEA treatment of intact starved-refed Sprague-Dawley rats resulted in significantly lower hepatic lipid and glucose-6-phosphate dehydrogenase activity than observed in non-DHEA-treated rats. When Sprague Dawley rats were adrenalectomized (ADX), the response to DHEA treatment was potentiated. If glucocorticoid was replaced, there was some amelioration of the DHEA effect in the ADX rats. Responses to DHEA in BHE rats subjected to the above paradigms were different. The responses of starved-refed BHE rats to DHEA were more pronounced and it appeared that glucocorticoid replacement was not as effective in overcoming DHEA in these rats. Thus, it appears that the comparative inhibition of the glucocorticoid-mediated response to starvation-refeeding by DHEA is strain dependent. PMID- 2969964 TI - [Permeability of round window membrane to human serum albumin in chinchillas]. PMID- 2969963 TI - Ethanol and leucine as possible stress factors in rats marginally deficient in niacin and vitamin B-6. AB - Young rats were fed a purified diet containing no niacin (and 0.85 g/kg tryptophan) and only 0.5 mg/kg vitamin B-6. Three supplements were tested in a 2 x 2 x 2 multifactorial experiment: 1) 25 mg/kg niacin plus 7.3 mg/kg vitamin B-6; 2) 15 g/kg L-leucine; and 3) 12.15% (wt/wt) ethanol in the drinking water. The ethanol, for those receiving it, contributed approximately 15% of the rats' total energy intake, and their diets had some of the carbohydrate omitted to maintain approximately the same proportion of energy to other nutrients. The vitamin supplements stimulated 4-wk weight gain (and gain-feed ratio), but ethanol depressed it, and to a significantly greater extent in the absence of the vitamin supplement. Excretion of N1-methylnicotinamide was increased only by the vitamin supplement. Leucine had no significant effects either by itself or by interaction with the other supplements. PMID- 2969965 TI - False positive rates in the determination of changes in probing depth-related periodontal measurements. PMID- 2969966 TI - Topical flurbiprofen treatment of periodontitis in beagles. PMID- 2969967 TI - Dark-pigmented Bacteroides species in subgingival plaque of adult patients on a rigorous recall program. PMID- 2969968 TI - Progenitor connective tissue cell populations in the gingival papilla of the rat. PMID- 2969969 TI - Immunohistochemical localization of epithelial cells expressing transferrin receptors in normal and diseased human dento-gingival junction. PMID- 2969970 TI - The effect of periodontal proteolytic Bacteroides species on proteins of the human complement system. PMID- 2969971 TI - Serum-antibodies against the trypsin-like protease of Bacteroides gingivalis in periodontitis. PMID- 2969972 TI - Black-pigmented Bacteroides and spirochetes in the subgingival microbiota of prepubertal schoolchildren. PMID- 2969973 TI - Long-term culture of human periodontal ligament cells with autologous root discs. PMID- 2969974 TI - In vitro antibacterial activity of ofloxacin against periodontal disease associated bacteria. PMID- 2969975 TI - Mechanism of ochratoxin A transport in kidney. AB - The effect of the fungal metabolite (mycotoxin) Ochratoxin A (OTA) on the transport of p-amino[3H]hippurate (PAH), a prototypic organic anion, was examined in renal brush border (BBMV) and basolateral membrane vesicles (BLMV). OTA was as effective an inhibitor of PAH uptake in both membranes as probenecid. The dose response curves for OTA in BBMV and BLMV gave IC50 values of 20 +/- 6 and 32 +/- 7 microM, respectively. The effect was specific since the transport of the organic cation N1-methylnicotinamide was not affected. The phenomenon of counterflow was studied to establish that OTA is translocated. OTA produced trans stimulation of PAH transport in both BBMV and BLMV, demonstrating that OTA is transported across both these membranes. The data suggest that OTA interacts with the PAH transport system in both BBMV and BLMV. We conclude that OTA transport in the kidney is mediated via the renal organic anion transport system. PMID- 2969976 TI - Blunted pressor responsiveness to quinpirole, a specific dopamine D2 receptor agonist, in conscious deoxycorticosterone acetate/NaCl hypertensive rats is related to atrial natriuretic peptide release. AB - Our previous studies have demonstrated: 1) that i.v. quinpirole (LY171555), a selective dopamine D2 receptor agonist, has a dose-dependent pressor effect in conscious rats which is mediated by activation of sympathetic outflow and vasopressinergic activity, and 2) that the activity of central dopaminergic neurons is reduced in deoxycorticosterone acetate (DOCA)/NaCl hypertensive rats. To elucidate the role of central and peripheral dopaminergic systems in the pathogenesis of DOCA/NaCl hypertension, we examined the effects of quinpirole on mean arterial pressure, heart rate, plasma norepinephrine, epinephrine, arginine vasopressin and atrial natriuretic peptide (ANP) in conscious 4-week-old DOCA/NaCl hypertensive and normotensive control rats. Quinpirole (1 mg/kg i.v.) increased mean arterial pressure in both groups, but the pressor response was attenuated in DOCA/NaCl rats. Paradoxically, quinpirole-induced increments in plasma norepinephrine, epinephrine and arginine vasopressin were greater in DOCA/NaCl rats. In addition, quinpirole induced a 2-fold increase in plasma ANP (P less than .01) in both DOCA/NaCl and control rats. Pretreatment with domperidone (2.5 mg/kg i.v.), a peripherally acting dopamine D2 antagonist, enhanced the maximum pressor response to quinpirole in both groups, restored the quinpirole-induced pressor response to control levels in the DOCA/NaCl rats and blocked the stimulatory effect of quinpirole on ANP release in both groups. These data indicate that peripheral dopamine D2 receptors modulate ANP secretion in the rat. The observation that the quinpirole-induced increment in plasma ANP was enhanced in DOCA/NaCl rats supports the hypothesis that the blunted pressor response to quinpirole in this model is related to enhanced ANP release. PMID- 2969977 TI - Atrial natriuretic peptide reverses afferent arteriolar vasoconstriction and potentiates efferent arteriolar vasoconstriction in the isolated perfused rat kidney. AB - The intrarenal sites of action of atrial natriuretic peptide (ANP) have not been resolved fully. Although ANP relaxes in vitro preparations of smooth muscle and exerts profound effects on renal hemodynamics, the effects of ANP on resistance vessels remain a subject of controversy. In the present study, we utilized an in vitro perfused hydronephrotic rat kidney model to assess directly the actions of ANP on renal microvessels during norepinephrine (NE)-induced renal vasoconstriction. Perfusion pressure was maintained constant, and perfusate flow to the kidney was monitored while the afferent arteriole (AA) and efferent arteriole (EA) were visualized using videomicroscopy. Renal AA and EA diameters were measured by computer-assisted image analysis. NE (0.3 microM) decreased renal perfusate flow from 11.8 +/- 1.7 (S.E.) to 5.8 +/- 1.2 ml/min (P less than .005), decreased AA diameter from 20.2 +/- 0.7 to 14.3 +/- 0.8 mu (P less than .005) and decreased EA diameter from 18.6 +/- 1.2 to 15.1 +/- 1.2 mu (P less than .005). ANP (human ANF-(4-28), anaritide, Wyeth) completely reversed the NE induced AA vasoconstriction with an IC50 of 9.3 +/- 5.6 nM. In contrast, ANP caused a further decrease in EA diameter at concentrations up to 10(-8) M, and elicited a slight dilation of the EA at 10(-7) M. In normal rat kidneys perfused under identical conditions, NE decreased renal perfusate flow, glomerular filtration rate and filtration fraction. ANP increased glomerular filtration rate and filtration fraction above control levels, with maximal effects at 10(-8) M.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969978 TI - Regional blood flows and cardiac function changes induced by angiotensin II in conscious dogs. AB - Hemodynamic properties of angiotensin (ANG) II 1, 5, 10 and 100 ng/kg i.v. and 10, 100 and 1000 ng/kg i.v.t. were assessed in conscious dogs. ANG II i.v. produced a dose-dependent pressor response (59 +/- 5-124 +/- 16 mmHg) and renal vasoconstriction (1.3 +/- 0.4-96 +/- 32 mmHg/ml/min). Ganglionic blockade (chlorisondamine 2 mg/kg i.v.) diminished mean arterial responses without altering peptide effects on renal circulation. At the highest dose, ANG II i.v. induced cardiac stimulation: increased heart rate (75 +/- 4-115 +/- 6 beats/min), cardiac output (2.0 +/- 0.1-2.4 +/- 0.2 l/min), dP/dt (2308 +/- 181-2773 +/- 173 mmHg/sec) and coronary blood flow (49 +/- 10-96 +/- 23 ml/min). Although with chlorisondamine cardiac response was more pronounced, subsequent beta blockade abolished it. Concomitantly, an isolated increase in plasma epinephrine was recorded (63 +/- 8-1505 +/- 354 pg/ml). A pressor response (59 +/- 8-89 +/- 13 mmHg) and renal vasoconstriction (1.1 +/- 0.1-2.2 +/- 0.5 mmHg/ml/min) were also produced by ANG II i.v.t. at the highest dose. These centrally mediated changes were prevented by chlorisondamine. Our study demonstrates 1) i.v. ANG II-mediated pressor responses are dependent on direct and indirect components, the relative contribution of each being dependent on the regional circulation; ANG II i.v. also produced a biphasic cardiac response--an initial centrally mediated depression and a secondary stimulation dependent on epinephrine via cardiac beta receptors and 2) i.v.t. ANG II-mediated pressor effects are essentially indirect. Finally, no evidence was found to support the role of vasopressin in ANG II effects. PMID- 2969979 TI - Natriuretic action of neurohypophysial peptides: effects of agonists and antagonists and implication of natriuretic receptor. AB - Neurohypophysial peptides possess natriuretic activity. Although it has been shown that the natriuretic action of these peptides can be dissociated from their antidiuretic activity (a V2-receptor mediated response), it is not known whether the V1-receptor or yet a third receptor type mediates the natriuretic response. Also, it has not been studied what effects V1- and V2-antagonists may have on urinary sodium excretion. To define this, we have studied the effects of four oxytocin (OT) agonists: arginine-vasopressin, OT, [Leu4]OT and [cyclo-Leu8]OT; two V1-receptor antagonists: [penicillamine1,Phe(Methyl)2,Thr4,Orn8]OT and [penicillamine1,D-Phe(Ethyl)2,Thr4,Orn8]OT and one V2-receptor antagonist: d (CH2)5[D-Ile2,alpha-aminobutyric acid4]arginine-vasopressin on renal excretion of water and electrolytes in anesthetized rats under water diuresis. We also studied the effects of the antagonists on the OT-induced antidiuretic and natriuretic responses. Only the agonists, but not the antagonists, were found to have natriuretic activity. The natriuretic potency was not related to the peptide's antidiuretic activity, but was in the same rank order as their oxytocic activity (a V1-agonist effect). The effects of the antagonists on the OT-induced renal responses were studied at two dose levels, representing a strong and near maximal of their respective V1 and V2 inhibitory doses. The V1-antagonist had no effect on the antidiuretic response to OT but inhibited the natriuretic response in a dose-dependent manner. The antinatriuretic effect was also long-lasting as its antioxytocic activity. The V2-antagonist inhibited the antidiuretic response to OT in a dose-dependent manner but only the high dose inhibited the natriuretic response. These results indicate that the natriuretic action of OT was not mediated by V2-receptors and antinatriuresis was not specific for V1 antagonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2969980 TI - The failed back: a review. PMID- 2969981 TI - Use of rapid survey methodology to determine immunization coverage in rural Burma. PMID- 2969982 TI - Expression of interferon-induced genes in different tissues of mice. AB - In vivo responses to interferon (IFN) in mice were determined by measuring the steady-state levels of induced mRNAs following injection of IFN and poly(I) poly(C). With cDNA probes for mouse 2'-5' oligoadenylate synthetase (2-5A synthetase) and 1-8, constitutive expression of the corresponding mRNA was detectable in different organs of normal C3H/He mice. These mRNA levels were increased by as much as 15-fold over control levels in various tissues, including the brain, after IFN and poly(I)-poly(C) treatment, coincident with increases in 2-5A synthetase enzyme activity. The basal activity level of this enzyme could be reduced in normal mice by treatment with anti-mouse IFN (alpha + beta) antibody. This treatment also reduced the levels of 2-5A synthetase and 1-8 mRNAs. Thus, physiological levels of circulating IFN maintain elevated levels of IFN-induced mRNAs in mice. Furthermore, changes in 2-5A synthetase enzyme activity reflect the changes in gene expression in vivo. PMID- 2969983 TI - Adenovirus type 12 E1B 19-kilodalton protein is not required for oncogenic transformation in rats. AB - The adenovirus type 12 mutants in700 and pm700 carry site-specific mutations within the reading frame encoding the E1B 19-kilodalton protein (19K protein) which prevent the production of the intact 19K protein. In cultures of human A549 cells, these mutants grow just as well as the wild-type virus does, but they display a large-plaque (lp), cytocidal (cyt) phenotype. DNA in these infected cells is not degraded, but at late times in human KB cells infected by the mutants, the mutants display a DNA degradation (deg) phenotype. The transformation phenotype of these mutants is also host range. Although the mutants are defective for transformation of the 3Y1 rat cell line, they transform rat and mouse primary kidney cells in vitro at wild-type efficiency and are capable of inducing tumors in rats. These results support the view that the type 12 E1B 19K protein is not obligatory for oncogenic transformation. PMID- 2969985 TI - [The significance of measurement of alpha 2PI-plasmin complex in acute promyelocytic leukemia]. PMID- 2969986 TI - [Clinical evaluation of alpha 2 plasmin inhibitor and alpha 2 plasmin inhibitor plasmin complex by enzyme immunoassay method--with special reference to accident surgery]. PMID- 2969987 TI - [Microchemistry by EIA]. PMID- 2969984 TI - Role of the adenovirus E1B 19,000-dalton tumor antigen in regulating early gene expression. AB - Mutations in the adenovirus gene encoding the E1B 19-kilodalton protein (the 19K protein) result in pleiotropic phenotypes that affect the host cell and virus growth. Examination of viral gene expression in HeLa cells infected with E1B 19K mutant viruses revealed synthesis and accumulation of E1A proteins to higher steady-state levels than those proteins synthesized during infection with the wild-type virus. As a consequence of elevated E1A levels, another early gene product, the 72K DNA-binding protein, accumulated earlier in mutant-infected cells. In a 12S E1A cDNA virus background, E1B 19K gene mutations had a more profound effect. Larger amounts of the 12S E1A product were present in E1B mutant infected cells. A deletion mutation that eliminated expression of the 19K protein was also responsible for a 200-fold increased plaque-forming efficiency of the 12S cDNA virus in HeLa cells and an increased rate of virus production. Therefore, the E1B 19K tumor antigen may function to down-regulate virus replication by repressing E1A-dependent gene transcription. Eliminating expression of the E1A 13S and 12S gene products by substitution of an E1A 9S cDNA gene, however, uncovered a stimulatory effect of the E1B 19K protein on early gene expression and virus replication. An E1A 9S virus with a wild-type gene encoding the E1B 19K protein displayed increased early gene transcription, synthesized more 72K DNA-binding protein, and replicated more efficiently than an E1A 9S virus containing a mutation that eliminated expression of the 19K protein. Therefore, the E1B 19K protein has both positive and negative effects on early gene expression and virus replication. In the presence of functional E1A gene products, the 19K protein repressed E1A-dependent gene expression, but in the absence of E1A, the 19K protein stimulated viral gene expression and DNA synthesis. This raises the possibility that the E1B 19K protein functions to repress transcription by modifying the activity of the E1A proteins. Independent of E1A, however, the E1B 19K protein can increase viral gene expression and DNA synthesis, which then leads to increased virus replication. PMID- 2969988 TI - [A study of drug eruptions due to tiopronin]. PMID- 2969989 TI - [A case of Hartnup disease]. PMID- 2969990 TI - The distribution of transferrin, group-specific component and phosphoglucomutase 1 subtypes among the Lepchas of Darjeeling, eastern India. PMID- 2969991 TI - Hemodialysis-associated subclavian vein stenosis. AB - This study was undertaken to evaluate hemodialysis-associated subclavian vein stenosis (SVS) and to clarify treatment of this condition. Forty-seven patients underwent upper arm venography to evaluate fistula dysfunction. Subclavian vein stenosis was documented in 12. Eleven of 12 had elevated venous dialysis pressure (196 +/- 8.9 mm Hg), and six had arm edema. All 12 had previously undergone subclavian cannulation on the side of the fistula. Thirty-five patients showed no evidence of subclavian vein stenosis. Twelve of these 35 patients (mean venous dialysis pressure 113 +/- 2.3 mm Hg) had undergone previous subclavian cannulation on the side of the fistula. The mean age of the fistula at the time of venogram in patients with subclavian vein stenosis was 17.0 months versus 5.8 months in patients with ipsilateral subclavian cannulation without subclavian vein stenosis. Percutaneous transluminal angioplasty (PTA) was performed on 11 of 12 patients with SVS lowering venous dialysis pressure and restoring patency to the fistula in 100%. Lesions recurred in two of 11 patients and were successfully retreated with PTA. We conclude that SVS is a common dialysis problem that is amenable to treatment with PTA. Elevated venous dialysis pressures are a sensitive indicator of this condition. PMID- 2969992 TI - [Diagnosis and treatment of open and closed abdominal injuries]. PMID- 2969993 TI - [Laparoscopy in the diagnosis of penetrating abdominal injuries]. PMID- 2969994 TI - [Hermetization of the abdominal wall wound after appendectomy]. PMID- 2969995 TI - [Is retinitis pigmentosa absolutely incurable?]. AB - The importance of phosphatides in the biochemistry of the retina and in visual function provided a basis for therapeutic administration of these substances in cases of senile maculopathy and retinitis pigmentosa. In cases of dry senile maculopathy, good results were only obtained by administering (n-3) docosahexaenacid to the retina as the active component of intramuscular injections of Etaretin. So far only one report of a convincing therapeutic success in a case of retinitis pigmentosa has been published. PMID- 2969996 TI - Role of T helper lymphocytes in autoimmune diseases. PMID- 2969997 TI - Effects of 2'-deoxy-5-fluorouridine on regenerating liver following partial hepatectomy in the rat. AB - This study evaluated the effect of 2'-deoxy-5-fluorouridine (FUDR) on the regeneration of the liver following partial (68%) hepatectomy in the rat. Male Sprague-Dawley rats weighing between 190 and 240 g underwent partial hepatectomy under ether anesthesia. Twelve hours postoperatively rats received intraperitoneal injections of 2'-deoxy-5-fluorouridine or 0.9% NaCl solution as follows: Group I, 0.9% NaCl solution (n = 49); Group II, 89 mg 2'-deoxy-5 fluorouridine/kg of body weight (n = 25); and Group III, 178 mg 2'-deoxy-5 fluorouridine/kg of body weight (n = 24). Sham groups underwent celiotomy and liver palpation followed by 0.9% NaCl solution injections (n = 5) or low dose 2' deoxy-5-fluorouridine (n = 5) and high dose 2'-deoxy-5-fluorouridine (n = 5). The regenerative ability of the liver was evaluated by weight and deoxyribonucleic acid synthesis in the liver remnant. RESULT: Both low and high dose 2'-deoxy-5 fluorouridine delayed the peak of deoxyribonucleic acid synthesis from 36 to 72 hr as compared to control animals which had maximal synthesis at 25 to 36 hr postoperatively (P less than 0.01). Weight of the liver remnants demonstrated a similar pattern. CONCLUSION: High doses of 2'-deoxy-5-fluorouridine administered intraperitoneally delay, but do not inhibit, liver regeneration following partial (68%) hepatectomy as reflected by DNA synthesis and weight of the remnant. PMID- 2969999 TI - Progestin regulation of protein synthesis in endometrial cancer. AB - Protein synthesis in cancerous and normal human endometrium was investigated by the incorporation of [35S]methionine and analysis of products by two-dimensional polyacrylamide gel electrophoresis. Comparison of the cellular products from organ cultures of endometrial carcinoma, obtained from each subject before and after in vivo administration of medroxyprogesterone acetate (MPA) for 10-14 days revealed: (i) a decrease in the synthesis of tubulin and of a protein of molecular weight (mol. wt) 68 kDa, isoelectric point (pI) 6.0, and (ii) an increase in the synthesis of creatine kinase (CK) and of protein of mol. wt 36 kDa, pI 4, following MPA therapy. The 68 kDa protein was expressed at relatively reduced levels in organ cultures of normal human endometrium throughout the menstrual cycle. In primary cultures of normal human endometrium throughout the menstrual cycle. In primary cultures from cancerous endometrium endometrium established for several weeks the 68 kDa protein was not expressed but could be induced by heatshock. Primary cultures were also used to investigate the early events following progesterone stimulation which revealed a decrease in synthesis of a protein mol. wt 36 kDa, pI 8 at 8 h following administration. PMID- 2969998 TI - Phasic coronary flow during aorto-caval fistula unclamping in dog. AB - The concomitant changes in components of external cardiac power and phasic coronary flow are described here in an aorto-caval fistula model (ACF) in the dog. Eleven animals were used in this study. The steady, oscillatory, and kinetic components of the external cardiac power and aortic impedance spectrum were calculated from the measurement of instantaneous flow and pressure in the ascending aorta. Arterial coronary flow was measured by a pulsed doppler flowmeter on the left anterior descending artery. The concomitant venous coronary flow was measured in the last two dogs. After unclamping of a large aorto caval fistula, mean blood pressure decreased from 96 +/- 31 mm Hg in the control state to 78 +/- 19 mm Hg 5 min after opening the shunt. Aortic blood flow and heart rate increased significantly. These changes in pressure and flow led to a decrease in the steady component of the external cardiac power (752 +/- 203 mW in the control state to 590 +/- 180 mW in the ACF; P less than 0.01), whereas the oscillatory and the kinetic components of the total external power increased (23.5 +/- 11 mW in the control state to 33 +/- 6 mW in the ACF for oscillatory power; 11.7 +/- 5.1 mW in the control state to 17.7 +/- 8.5 mW in the ACF for kinetic power, P less than 0.05). This increase in the oscillatory component of the cardiac power corresponded to the increase in the characteristic impedance of the aorta in the ACF (2,507 +/- 231 dyn.sec.cm-5 in the control state versus 4,366 +/- 745 dyn.sec.cm-5 in the ACF). These results suggested poor energetic efficiency in the matching between the left ventricle and the large arteries in this model. After unclamping the fistula, the systolic component of arterial coronary inflow becomes biphasic with a constant reverse flow from the myocardium to the aorta. The diastolic part of coronary arterial inflow decreased markedly, secondary to the decrease in perfusion pressure, and then reincreased to values near control values (59 +/- 22 ml/min in the control state versus 52 +/- 18 ml/min 5 min after unclamping the shunt). In five cases, the abrupt fall in blood pressure induced by unclamping the fistula led to a complete disappearance of diastolic coronary arterial inflow in diastoles of normal duration. The measured pressure of this zero inflow was about 30 mm Hg. During this zero coronary inflow, venous coronary outflow decreased but never disappeared.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970001 TI - [Cutaneous reactions of atracurium]. PMID- 2970000 TI - The effect of short term treatment with cyproterone acetate or flutamide on the metabolism of 5 alpha-dihydrotestosterone in human testicular tissue. AB - Testicular tissue obtained from ten patients orchiectomized for prostatic cancer was incubated with [3H]5 alpha-dihydrotestosterone (DHT) in order to study the metabolic transformation into 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha diol) and 5 alpha-androstane-3 beta,17 beta-diol (3 beta-diol). Throughout 5 days before surgery four subjects were treated with cyproterone acetate (CA). To three patients flutamide (F) was administered for the same period of time. Three subjects remained untreated. Compared to the control group the administration of CA decreased the formation of 3 beta-diol whereas that of 3 alpha-diol increased. Treatment with F lead to an elevated formation of both diols. However, the 3 alpha/3 beta ratio did not change. As 3 beta-diol is considered to be an index of tubular function in the human testis it is concluded that CA has a direct inhibitory effect upon this testicular compartment whereas F has none. PMID- 2970002 TI - [Baastrup's disease of the lumbar segment of the spine among drivers of heavy motor vehicles]. AB - The strain of the lower part of spine in professional work is a complex social problem. The problem is particularly enhanced in highly industrialized countries. Many strain pains are related to the Baastrup disease. Analysed is the prevalence rate of this disease in a group of heavy automotive vehicles drivers, which constituted 13% of the test population. The obtained results were compared with a control group of automotive mechanical engineers and subsequently verified statistically, which indicated a higher prevalence rate of the Baastrup disease in professional drivers. Most frequently the test material revealed lesions of the spine L3-L4 and L4-L5 segments. It was decided advisable to undertake extensive preventive measures related to the strain of the lower part of spine in the working population. PMID- 2970003 TI - Effects of TRIS and HEPES on function of rabbit muscle light sarcoplasmic reticulum. AB - ATP hydrolysis activity and calcium transport activity were determined on light sarcoplasmic reticulum from rabbit skeletal muscle. The effects of two buffers, TRIS and HEPES, were compared. Titration of TRIS into sarcoplasmic reticulum preparations in HEPES provided evidence for TRIS inhibition of ATPase activity and TRIS stimulation of calcium transport activity. PMID- 2970005 TI - [Renovascular hypertension]. PMID- 2970004 TI - Mechanism of the stimulation of cardiac sarcoplasmic reticulum calcium pump by calmodulin. AB - Calmodulin has been shown to stimulate the initial rates of Ca2+-uptake and Ca2+ ATPase in cardiac sarcoplasmic reticulum, when it is present in the reaction assay media for these activities. To determine whether the stimulatory effect of calmodulin is mediated directly through its interaction with the Ca2+-ATPase, or indirectly through phosphorylation of phospholamban by an endogenous protein kinase, two approaches were taken in the present study. In the first approach, the effects of calmodulin were studied on a Ca2+-ATPase preparation, isolated from cardiac sarcoplasmic reticulum, which was essentially free of phospholamban. The enzyme was preincubated with various concentrations of calmodulin at 0 degrees C and 37 degrees C, but there was no effect on the Ca2+-ATPase activity assayed over a wide range of [Ca2+] (0.1-10 microM). In the second approach, cardiac sarcoplasmic reticulum vesicles were prephosphorylated by an endogenous protein kinase in the presence of calmodulin. Phosphorylation occurred predominantly on phospholamban, an oligomeric proteolipid. The sarcoplasmic reticulum vesicles were washed prior to assaying for Ca2+ uptake and Ca2+-ATPase activity in order to remove the added calmodulin. Phosphorylation of phospholamban enhanced the initial rates of Ca2+-uptake and Ca2+-ATPase, and this stimulation was associated with an increase in the affinity of the Ca2+-pump for calcium. The EC50 values for calcium activation of Ca2+-uptake and Ca2+-ATPase were 0.96 +/- 0.03 microM and 0.96 +/- 0.1 microM calcium by control vesicles, respectively. Phosphorylation decreased these values to 0.64 +/- 0.12 microM calcium for Ca2+-uptake and 0.62 +/- 0.11 microM calcium for Ca2+-ATPase. The stimulatory effect was associated with increases in the apparent initial rates of formation and decomposition of the phosphorylated intermediate of the Ca2+ ATPase. These findings suggest that calmodulin regulates cardiac sarcoplasmic reticulum function by protein kinase-mediated phosphorylation of phospholamban. PMID- 2970006 TI - [Study on splanchnic circulation: measurement of the liver blood flow]. AB - In anesthetized patients during abdominal surgery, hepatic artery and portal vein flows were measured simultaneously utilizing an ultrasonic transit-time volume flowmeter. The total hepatic blood flow was 994.6 +/- 52.4 ml/min. The hepatic artery flow and the portal vein flow were 260.0 +/- 23.8 ml/min and 730.8 +/- 41.3 ml/min, respectively. The ratio of hepatic artery flow to portal vein flow was 0.37 +/- 0.04. A significant increase in hepatic artery flow (p less than 0.01) followed portal vein occlusion, whereas no significant change was observed in portal vein flow after hepatic artery occlusion. Common hepatic artery occlusion resulted in a significant decrease in hepatic artery flow (p less than 0.05), but no significant change was observed in portal vein flow. The present study firstly demonstrated that ultrasonic transit-time volume flowmeter is a device to quantitatively assess hepatic artery and portal vein flows with good reproducibility and stability in human subjects. This easy and simple technique seemed to have wide clinical application to abdominal surgery and would have a promising in studying splanchnic blood flows in various situations such as in cases of hepatectomy and portal hypertension. PMID- 2970007 TI - Molecular analysis of a Neurospora crassa gene expressed during conidiation. AB - The asexual developmental pathway in the life cycle of the filamentous fungus Neurospora crassa culminates in the formation of spores called conidia. Several clones of genomic Neurospora DNA have been isolated that correspond to mRNA species expressed during conidiation and not during mycelial growth (V. Berlin and C. Yanofsky, Mol. Cell. Biol. 5:849-855, 1985). In this paper we describe the characterization of one of these clones, named pCon-10a. This clone contains two genes, con-10 and con-13, which are induced coordinately during the later stages of conidiation. The two genes are separated by 1.4 kilobases of DNA; they are located on linkage group IV and are transcribed from the same strand of DNA. The molecular organization and sequence of one of these genes, con-10, and its flanking regions are presented. Full-length cDNA clones for con-10 also were isolated and sequenced, and transcription-initiation and polyadenylation sites were defined. The con-10 gene contains an open reading frame interrupted by two small introns and encodes an 86-amino-acid residue polypeptide that is both hydrophilic and weakly acidic. Expression of the con-10 gene in various mutants defective at different stages of conidiation indicates that it plays a role after aerial hyphal development. Possible functions, organization, and regulation of conidiation-specific genes are discussed. PMID- 2970008 TI - The E1a gene of adenovirus type 2 reduces the metastatic potential of ras transformed rat embryo cells. AB - We have previously demonstrated that second-passage rat embryo cells transformed by the ras oncogene alone are both tumorigenic and highly metastatic when injected into nude mice. In contrast, rat embryo cells cotransformed with the ras oncogene and the adenovirus type 2 (Ad2) E1a gene are tumorigenic but either fail to metastasize or exhibit a very low metastatic potential. In this report, we demonstrate that transfection of the Ad2 E1a gene into four independent ras transformed rat embryo cell lines results in a dramatic reduction in metastatic potential relative to that of the parental cell line. Transfection of cDNAs for the 12S and 13S E1a transcripts showed that the 12S cDNA was highly effective in reducing the metastatic potential of ras-transformed cell lines, while the 13S cDNA showed an effect in only one of the two cell lines tested. This effect is specific to the Ad2 E1a gene, since ras-transformed cell lines expressing the Ad12 E1a gene maintained their high metastatic potential. We hypothesize that the Ad2 E1a gene may regulate the expression of one or more cellular genes that contribute to the metastatic phenotype. PMID- 2970009 TI - Ipratropium bromide. PMID- 2970010 TI - Nafarelin for treatment of endometriosis. PMID- 2970011 TI - Predictive testing for Huntington's disease using linked DNA markers. PMID- 2970012 TI - The role of beta-endorphin in respiratory disorders in man. PMID- 2970013 TI - Ribosomal RNA sequence shows Pneumocystis carinii to be a member of the fungi. AB - Pneumocystis carinii pneumonia is the most common opportunistic infection in AIDS, and accounts for significant morbidity and mortality in these and other immunocompromised patients. P. carinii is a eukaryotic microorganism of uncertain taxonomy that can infect numerous mammalian hosts. Developing from a small, unicellular 'trophozoite' into a 'cyst' containing eight 'sporozoites', its life cycle superficially resembles those seen both in the Protozoa and Fungi. Morphological and ultrastructural observations have lead some investigators to conclude that the organism is a protozoan, while others have felt that it more closely resembles a fungus. Phylogenetic relationships can be inferred from comparisons of macromolecular sequences. Small subunit ribosomal RNAs (16S-like rRNAs) are well-suited for this purpose because they have the same function in all organisms and contain sufficient information to estimate both close and distant evolutionary relationships. Phylogenetic frameworks based upon such comparisons reveal that the plant, animal and fungal lineages are distinct from the diverse spectrum of protozoan lineages. In this letter, phylogenetic analysis of Pneumocystis 16S-like rRNA demonstrates it to be a member of the Fungi. PMID- 2970015 TI - [Synovial cysts of the lumbar interapophyseal joints. Apropos of 2 cases and review of the literature]. AB - The authors report two cases of low back pains with sciatica caused by a synovial cyst of the interapophyseal joint of the lumbar spine. Analysis of only 23 cases found in the literature, point to their rarity, the lack of specific clinical signs, the importance of the CT scan for the diagnosis and the excellent results of surgical treatment. PMID- 2970014 TI - The significance of extracellular calcium for the release of dopamine, acetylcholine and amino acids in conscious rats, evaluated by brain microdialysis. AB - The influence of the calcium concentration of the perfusion fluid on the release of striatal dopamine recorded by brain dialysis was investigated. The release of dopamine appeared very sensitive to the calcium concentration of the Ringer. Next we studied whether three different methods known to antagonize the effects of calcium entry, were able to affect the release of dopamine. The conditions investigated were: the use of calcium-free Ringer, infusion of magnesium, and infusion of the calcium-antagonist verapamil. Calcium-antagonism was studied on the day of implantation of the cannula as well as on several days thereafter. It appeared that magnesium infusion was the most effective condition to antagonize the effects of calcium on the release of dopamine. Magnesium infusion was also most effective in preventing drug-evoked voltage-dependent dopamine release (induced by coadministration of haloperidol and GBR 12909). In addition magnesium infusion appeared a potent antagonist of acetylcholine release. In contrast, the dialysate content of aminoacids was not influenced by magnesium infusion. PMID- 2970016 TI - Evidence for altered methionine methyl-group utilization in the diabetic rat's brain. AB - The methionine (MET) derivative, S-adenosylmethionine (SAM), provides methyl groups for methylation reactions in many neural processes. In rats made diabetic with streptozotocin (SZ), brain SAM levels were generally lower (10-20%) than in controls, with a constant decrease being observed five weeks after onset of diabetes. This decrease in SAM levels may be due to reduced precursor (MET) availability because greatly elevating plasma MET concentrations in SZ diabetic rats by dietary manipulation increased their neural SAM concentrations to be approximately or even greater than (5-20%) those of controls. In contrast, neural levels of SAM's demethylated product, S-adenosylhomocysteine (SAH), were reduced to a greater extent (17-44%) than SAM levels in all groups of SZ diabetic rats independent of their plasma MET concentrations or brain SAM levels. This indicates that the decrease in SAH levels is not simply due to substrate (SAM) restriction. These changes in MET metabolites appear to be a general effect of diabetes rather than a non-pancreatic side-effect of SZ, because genetically diabetic BB Wistar rats also exhibited reduced brain SAM (25%) and brain SAH (46%) levels. These results indicate that methyl-groups from MET are handled differently in the brain of the diabetic rat, which considering the variety and importance of neural methylation reactions, could have important consequences for the diabetic. PMID- 2970017 TI - Differential patterns of local cerebral glucose utilization in response to 5 hydroxytryptamine agonists. AB - Local cerebral glucose utilization was measured in parallel groups of conscious rats following intravenous injection of either 1 mg/kg 8-hydroxy-2-(di-N propylamino)tetralin (a 5-hydroxytryptamine 1A binding site agonist), 3 mg/kg 5 methoxy 3-(1,2,3,6-tetrahydro-4-pyridinyl)1H indole, succinate (a 5 hydroxytryptamine1B agonist), or saline alone, using the 2-deoxyglucose quantitative autoradiographic techniques (n = 5 in each of the three groups). Following both drugs, local rates of glucose use in the majority of the 72 brain areas analysed remained unaltered, but in some other regions either increases or decreases were observed. In keeping with the observed behavioural response to 8 hydroxy-2-(di-N-propylamino)tetralin there were marked increases in cerebellum (+56%) and motor cortex where a columnar arrangement of increased metabolism (+34%) contrasted with adjacent columns of decrease (-26%). Hippocampal areas showed moderate decreases in glucose use (-13 to -21%). All areas which increased following 8-hydroxy-2-(di-N-propylamino)tetralin also increased following 5 methoxy 3-(1,2,3,6-tetrahydro-4-pyridinyl)1H indole, succinate, but in the latter case all elements of the basal ganglia were also increased, including globus pallidus (+105%) and the striatum where the changes (+54%) were limited to a discrete dorsal region of the nucleus. In the hippocampus only dorsal dentate gyrus was decreased (-24%) whilst a moderate increase (+16%) was observed in dorsal subiculum. The complexity of these results contrasts with previous 2 deoxyglucose investigations where less specific 5-hydroxytryptamine receptor ligands were used, and suggests that certain aspects of brain function may be selectively targeted by systemic pharmacological manipulation of endogenous serotonergic systems. PMID- 2970018 TI - Stage-salient issues: a transactional model of intervention. PMID- 2970019 TI - [Critical review on the validity of Doppler velocimetry in the instrumental diagnosis of TIA (transient ischemic attack) and RIA (reversible ischemic attack)]. PMID- 2970020 TI - [Kinking and coiling of the carotid vessels in relation to ischemic cerebrovascular pathology]. PMID- 2970021 TI - [Organic and functional vertebro-basilar insufficiency. Our experience]. PMID- 2970022 TI - [Subclavian steal, Doppler velocimetry and digital angiography]. PMID- 2970023 TI - [Arteritis caused by immune complexes]. PMID- 2970024 TI - Hepatitis B vaccination in Tauranga. PMID- 2970025 TI - Hepatitis B immunisation. PMID- 2970027 TI - The mechanical mule: can you help? PMID- 2970026 TI - Acne and minocycline. PMID- 2970028 TI - A multi-disciplinary approach to psychopaedic nursing. PMID- 2970029 TI - Evaluation of oral medroxyprogesterone acetate in the treatment of endometriosis. AB - Medroxyprogesterone acetate was administered orally at the daily dose of 50 mg for four months to 21 symptomatic women with moderate to severe endometriosis, staged according to the American Fertility Society classification. The efficacy of the therapy was evaluated prospectively by the patients' symptomatology, monthly pelvic examinations, and by restaging the disease at second-look laparoscopy during the last week of treatment. The effects of therapy on the serum gonadotropin and ovarian steroid levels and on the endometrium and the implants of endometriosis were also evaluated. Improvement of symptoms, pelvic nodularity, and tenderness occurred in 80% of the patients. The mean stage score of disease by the American Fertility Society classification decreased from 18.2 +/- 2 before therapy to 5.9 +/- 1 after therapy (P less than .005). Amenorrhea, breakthrough bleeding, and persistent cyclic bleeding occurred in 75, 20, and 10% of the patients, respectively. Ovulation was inhibited in all patients, but serum hormone changes were statistically significant only for LH, which decreased from 9.5 +/- 4 to 5.2 +/- 2 IU/L (P less than .001), and estradiol (E2), which decreased from 80 +/- 30 to 46 +/- 26 pg/mL (P less than .02). Atrophic changes and pseudodecidualized reaction occurred in both the endometrium and the implants of endometriosis. In summary, oral medroxyprogesterone acetate is effective in relieving symptoms and objectively improving endometriosis. Besides inducing a pseudodecidualized reaction and atrophic changes in the endometrium and ectopic implants, medroxyprogesterone acetate also suppresses ovulation and the serum levels of both LH and E2. PMID- 2970030 TI - Antenatal diagnosis of giant cystic cavernous hemangioma by Doppler velocimetry. AB - A fetal extracranial mass seen on ultrasound was found by Doppler velocimetry to have low resistance to blood flow through its channels. The antenatal diagnosis of giant cavernous hemangioma was confirmed after birth. PMID- 2970031 TI - Laparoscopic management of fallopian tube prolapse. AB - Sporadic cases of fallopian tube prolapse and various methods of management have been reported since the initial case described in 1902. Two cases were managed recently by a combined vaginal and laparoscopic approach. Total salpingectomy was accomplished with minimal difficulty and limited invasiveness. A brief summary of each case and detailed description of the operative technique are presented. PMID- 2970032 TI - [Continuing development of the Public Health Service--assessment of health policy and scientific literature]. PMID- 2970033 TI - [Episodes of smog in North Rhine Westphalia and their effects on health]. PMID- 2970034 TI - [The effects of episodes of smog on mortality]. PMID- 2970035 TI - [[Noxious air-borne particles and respiratory diseases in children]. PMID- 2970036 TI - [Disease-induced impairment of living conditions in the population of Munich]. PMID- 2970038 TI - [Regional differentiation among expenditures of general local health insurance for drugs]. PMID- 2970037 TI - [Methodological questions on environment analysis--on the predictive value of environmental toxicological questions]. PMID- 2970039 TI - [Implementation of the GMK (Health Ministry) resolution "Health education and public health service"]. PMID- 2970040 TI - [Rubella vaccination--an ever-current theme]. PMID- 2970041 TI - [Historical development of occupational protection for women]. PMID- 2970042 TI - [Women industrial workers as the target group of health programs in the 20's]. PMID- 2970043 TI - [Women in employment. Social and occupational medicine aspects. The concept of the total project]. PMID- 2970044 TI - [The professional image of the nurse--a part of the history of female employment]. PMID- 2970045 TI - [Stresses and responsibilities in nursing care]. PMID- 2970046 TI - [Stresses and responsibilities of women workers in the textile and clothing industry]. PMID- 2970048 TI - [Female employment in English language research--change in paradigms in social and occupational medicine?]. PMID- 2970047 TI - [Women at the data terminal--current status]. PMID- 2970049 TI - [History of female employment]. PMID- 2970050 TI - [Reasons for disability in female workers in comparison to male workers in Wurttembern 1900-1940]. PMID- 2970051 TI - [Female employment and the development of pensions. 1]. PMID- 2970052 TI - [Female employment and the development of pensions. 2]. PMID- 2970053 TI - Total joint replacement: a consideration for antimicrobial prophylaxis. AB - Infection is the principal and most devastating complication of total joint replacement, resulting in long periods of hospitalization, staggering costs, loss of the implant, disastrous physical impairment, and even death. Staphylococcus aureus and Staphylococcus epidermidis account for more than 50% of late infections. Animal studies have shown that joint implants are at a high risk of becoming infected via a metastatic hematogenous route during transient bacteremias. Because cephalosporins have been established as the perioperative and intraoperative agents of choice to prevent infections related to total joint replacement, oral cephalosporins are the drugs of choice to minimize the potential for the metastatic infection of prosthetic joints associated with transient dental bacteremias. Clindamycin is preferred for patients who are allergic to the cephalosporins or who may have a cross-allergy between penicillin and the cephalosporins. PMID- 2970054 TI - Dental management of the patient with hemophilia. AB - Classic hemophilia is the most common inherited bleeding diathesis. Recent advances in the management of hemophilia have enabled many hemophiliac patients to receive outpatient dental care on a routine basis. An overview of hemophilia, including etiology, clinical features, and recent advances in treatment, is presented. Guidelines for dental management of the hemophiliac patient are discussed. PMID- 2970055 TI - [Therapeutic possibilities for the handicapped child]. AB - This article reviews the papers on therapies for handicapped children which were delivered at the 20th annual "Fortbildungskurs der Osterreichischen Gesellschaft fur Kinderheilkunde", Obergurgl, 1987. Central topic was the oral region. J. P. Guggenbichler, Innsbruck (pathogenesis) and Sieglinde Zschiesche, Erlangen (The Erlangen interdisciplinary treatment approach) covered genesis and treatment of cheilo-gnatho-palato-schisis. Background and therapy-principles for dysfunctions of the facial, oral and pharyngeal region were discussed by H. Haberfellner, Innsbruck. He also described therapy by use of modified orthodontic devices (ISMAR: Innsbruck sensori-motor activators and regulators) and Schonherr's vestibular screen. The same was done for Castillo-Morales' palatal plates by J. Limbrock, Munchen. Essence and changing features were described for Bobath concept and Peto's Conductive Education by H. Haberfellner and for Vojta's and Castillo-Morales' therapy by J. Limbrock. This part was concluded by a survey on therapeutic sports (H. Haberfellner): Riding on horseback, swimming, skiing and cross-country skiing, also for spinally injured patients. PMID- 2970056 TI - Difference in analgesia following epidural blockade in patients with postoperative or chronic low back pain. AB - Subjective responses of continuous epidural analgesia with bupivacaine were compared in 30 patients with acute (postoperative) or chronic (low back) pain. In the acute pain patients, sensory block was 4 dermatomes at 9 h and 6 dermatomes at 64 h. Corresponding values in the chronic pain patients were 8 and 6 dermatomes respectively. Motor blockade of the lower limbs was more profound in the acute pain group. The acute pain patients had significantly better pain relief (VAS: 85-96% vs. 55-70%) and a significantly higher proportion of these patients reported a global score of 3 (excellent; 80% vs. 7%). The mean dosage of bupivacaine decreased in the acute pain group from 21.0 +/- 5.7 (mean +/- S.D.) mg/h at 9 h to 15.1 +/- 8.5 mg/h at 64 h. Corresponding values for the chronic pain group were 20.7 +/- 5.9 and 12.0 +/- 6.0 mg/h respectively. Mean plasma concentration of bupivacaine increased from 1.2 +/- 0.8 micrograms/ml at 9 h to 2.1 +/- 1.4 micrograms/ml at 64 h in the acute pain patients and was 0.8 +/- 0.3 micrograms/ml at 9 h to 1.0 +/- 1.0 micrograms/ml at 64 h in the chronic pain patients. The incidence of side effects was approximately the same in both groups. No signs of accumulation or toxic reactions to bupivacaine were seen. PMID- 2970057 TI - Neuromuscular thermography in orthopaedic surgery. A usage poll. AB - The value of thermography in evaluating neck and back pain is assessed based on a simple random sample of 405 Active Fellows of the American Academy of Orthopaedic Surgeons. There were 316 responses to the survey with 293 Fellows indicating that they evaluate neck and back pain. Eighteen use thermography and seven find thermography helpful in their practice. The use of thermography in the courtroom to document painful conditions of the neck and back is not supported by this survey. PMID- 2970058 TI - [Efficacy of teicoplanin in 25 cases of severe infections caused by gram-positive cocci]. AB - Teicoplanin was evaluated in 25 severe infections caused by Gram-positive bacteria alone [13], in combination [9], alone then in combination [3], in an open study. Of 22 infections that could be evaluated, there were 18 cures and improvements (81.8%), 2 recurrence and 2 failures. Serum concentrations are unpredictable and steady after 5 days i.v. teicoplanin. Trough and peak concentrations are respectively less than 5 mg/l and less than 20 mg/l in 13 infections and higher in 9, unrelated with success or failure. A trough concentration greater than CMI and kind of infection seem deciding. The choice of the antibiotic combined with teicoplanin warrant more data. PMID- 2970059 TI - Carbamazepine: an alternative drug for the treatment of nonhereditary chorea. AB - Five patients with a moderate to severe degree of nonhereditary chorea were successfully treated with standard anticonvulsant doses of carbamazepine. In two cases, the cause of chorea was related to a streptococcal infection. In another patient, the involuntary movements appeared seven days after severe head injury. The cause in the remaining two patients could not be determined with certainty, despite extensive laboratory investigations. Improvement began within four to 15 days after the initiation of therapy. No side effects were noticed in four patients throughout treatment (3 months to 36 months). In one patient, the medication had to be discontinued after 17 days, because of an allergic cutaneous rash. In view of our results, we propose that carbamazepine be considered as an alternative drug for the treatment of nonhereditary chorea. PMID- 2970060 TI - An intermediate in the phage lambda site-specific recombination reaction is revealed by phosphorothioate substitution in DNA. AB - It has been proposed that phage lambda site-specific recombination proceeds via two independent strand exchanges: the first exchange forming a Holliday-structure which is then converted into complete recombinant products by the second strand exchange. If this hypothesis is correct, one should be able to trap the putative Holliday intermediate by preventing the second strand exchange. In this paper, we show that substitution of phosphorothioate for phosphate in one strand of a recombination site is an effective way to block recombination while permitting the accumulation of a novel structure. This effect is seen only when phosphorothioate is positioned at a point of potential cleavage by Int recombinase, demonstrating that the inhibition of strand exchange is highly specific. Analysis of the novel structure that accumulates in these reactions proves that it contains a Holliday joint. Holliday-structures can also be detected in unblocked recombinations but are present at very low levels. The characteristics of Holliday-structure formation that we describe substantiate the proposed recombination pathway. PMID- 2970062 TI - Structural analysis of the waxy locus from Hordeum vulgare. PMID- 2970063 TI - Rebels with a cause. PMID- 2970061 TI - The Saccharomyces cerevisiae RAD18 gene encodes a protein that contains potential zinc finger domains for nucleic acid binding and a putative nucleotide binding sequence. AB - The RAD18 gene of Saccharomyces cerevisiae is required for postreplication repair of UV damaged DNA. We have isolated the RAD18 gene, determined its nucleotide sequence and examined if deletion mutations of this gene show different or more pronounced phenotypic effects than the previously described point mutations. The RAD18 gene open reading frame encodes a protein of 487 amino acids, with a calculated molecular weight of 55,512. The RAD18 protein contains three potential zinc finger domains for nucleic acid binding, and a putative nucleotide binding sequence that is present in many proteins that bind and hydrolyze ATP. The DNA binding and nucleotide binding activities could enable the RAD18 protein to bind damaged sites in the template DNA with high affinity. Alternatively, or in addition, RAD18 protein may be a transcriptional regulator. The rad18 deletion mutation resembles the previously described point mutations in its effects on viability, DNA repair, UV mutagenesis, and sporulation. PMID- 2970064 TI - Differential diagnosis of seborrheic dermatitis. PMID- 2970065 TI - Metabolic changes in pectoral muscle and liver of turkey embryos in relation to hatching: influence of glucose and antibiotic-treatment of eggs. AB - Small type turkey eggs were pressure differential dipped in an antibiotic solution (spectinomycin 3,000 ppm) with or without glucose (10%) and incubated. Pectoral muscle and liver samples taken from the embryos at the knocking, pipping, and posthatch stages of development were studied. Muscle glycogen decreased between pipping and posthatch. Liver glycogen decreased progressively between knocking and posthatch. Whereas liver free fatty acids decreased as the embryo completed the hatching process, muscle levels were less subject to change. Providing the embryo with antibiotic plus glucose improved overall glycogen in both tissues and reduced muscle lactate. Results from treatment with antibiotic alone resembled those in the first treatment with regard to liver glycogen and muscle lactate but to a lesser degree. Results support the hypothesis of an extensive muscle involvement in the hatching process and need for glucose through the perihatch period. PMID- 2970066 TI - [General anesthesia in patients with various pediatric cardiopathies]. PMID- 2970067 TI - [Translocation of permanent teeth in Down's syndrome]. PMID- 2970068 TI - [Dental practice and arterial hypertension]. PMID- 2970069 TI - [Dental practice and heart diseases]. PMID- 2970070 TI - [Diabetes mellitus and dental practice]. PMID- 2970071 TI - [The renal and hepatic system]. PMID- 2970072 TI - [Antibiotic prophylaxis in dentistry (emphasis on prevention of infectious endocarditis)]. PMID- 2970073 TI - [Dental care for the patient with personality disorders]. PMID- 2970074 TI - [Prevention of infectious diseases]. PMID- 2970075 TI - [Hemorrhagic diseases and their emergency treatment]. PMID- 2970076 TI - [The significance of grief for the acceptance of a child's handicap]. PMID- 2970077 TI - [Protein C: importance of its assay in pathology]. PMID- 2970078 TI - [Portal hypertension in adults caused by chronic non-cirrhotic hepatopathies]. PMID- 2970079 TI - [Myxedematous coma. Prognostic and therapeutic re-evaluation]. AB - The prognostic factors and therapeutic approaches in myxoedema coma--a rare but serious medical emergency--were re-evaluated from a retrospective study of 10 cases. The immediate respiratory risk can be prevented by referring these patients immediately to an intensive care unit. The patient's cardiovascular status before and after coma determines the prognosis for life and serves as guideline to treatment. Hormone replacement therapy with initial injection of a loading dose of thyroxine seems to improve the prognosis by ensuring rapid recovery of the principal vital functions. PMID- 2970080 TI - [Prevention of peritonitis during continuous ambulatory peritoneal dialysis. Value of disconnectable systems]. AB - Peritonitis remains the major obstacle to the acceptance of continuous ambulatory peritoneal dialysis as a long-term dialysis technique. In January, 1985, Y connectors were introduced into our continuous ambulatory peritoneal dialysis programme, and a two-year prospective randomized trial for all new patients was initiated in which the Y connection system was compared with the conventional technique in the prevention of peritonitis (group I). At the same time, 16 patients (group II), with a high incidence of peritonitis episodes were switched from the conventional technique to the Y connection system, while 55 patients (group III), remained on the conventional technique. Group IA patients (27 new patients using the Y connection system), developed peritonitis every 23 patient months. Group IB patients (28 new patients using the conventional technique), developed peritonitis every 12.2 patient-months. The difference between these two sub-groups was statistically significant (P less than 0.02). Before their transfer to the Y connection system, group II patients developed peritonitis every 10 patient-months and thereafter one every 24 patient-months (P less than 0.001). Group III patients were divided into 12 continuous cyclic peritoneal dialysis patients with peritonitis every 24 patients-months, and 43 continuous ambulatory peritoneal dialysis patients with peritonitis every 11.7 patient months. The Y connector therefore proved to be a simple and safe procedure effective in reducing the peritonitis rate in patients on continuous ambulatory peritoneal dialysis. PMID- 2970081 TI - [Acanthosis nigricans, hyperandrogenism, insulin resistance and mixed hyperlipemia]. AB - We report the case of a 22-year old woman who presented skin lesions of acanthosis nigricans, hirsutism and secondary amenorrhoea. She had high plasma levels of adrenal androgens and low plasma levels of sex steroid binding protein. Polycystic ovaries were discovered in the course of a laparotomy performed for paraovarian cyst. An oral glucose tolerance test revealed a state of hyperinsulinism with intolerance to carbohydrates, while the body mass index was normal. This insulin resistant state corresponded in vitro to a decrease in the number of erythrocyte insulin receptors without decrease in their affinity for insulin. Following paradoxical improvement during a full-term pregnancy, there was gradual deterioration of diabetes control requiring insulin therapy. This metabolic decompensation was accompanied by major hyperlipaemia followed by acute haemorrhagic pancreatitis. This case illustrates the course of a type A insulin resistance syndrome which was detected at an early stage in front of an hirsutism acanthosis nigricans association. The underlying pathogenic mechanisms of these pathologies are discussed. PMID- 2970082 TI - [An uncommon aspect of medullary carcinoma of the thyroid gland: 2 cases]. PMID- 2970083 TI - [Plasmodium vivax malaria on the Tunisian coast]. PMID- 2970084 TI - [Value of Ringer lactate in the treatment of cibenzoline poisoning]. PMID- 2970085 TI - [Extrinsic allergic alveolitis manifested by acute respiratory distress syndrome]. PMID- 2970086 TI - [Thrombosis of a mitral valve prosthesis in thrombopenia induced by heparin. Fibrinolytic treatment]. PMID- 2970087 TI - [Acquired multicystic disease of dialysed patients. Incidence and complications]. PMID- 2970088 TI - [Vesical neuropathy in acute disseminated lupus erythematosus. 2 cases]. PMID- 2970089 TI - [Bullous pemphigoid following graft-versus-host reaction after bone marrow allograft]. PMID- 2970090 TI - [Value of magnetic resonance imaging after severe hypoglycemic coma]. PMID- 2970091 TI - [Chloroquinoresistant malaria after coming from Ghana]. PMID- 2970092 TI - [Definition of microalbuminuria]. PMID- 2970093 TI - Therapeutic effects of dehydroepiandrosterone (DHEA) and its metabolites in obese hyperglycemic mutant mice. AB - Dehydroepiandrosterone (DHEA) fed at 0.4%, and its metabolites, 3 alpha hydroxyetiocholanolone (alpha-ET) and 3 beta-hydroxyetiocholanolone (beta-ET), fed at 0.1%, had marked anti-hyperglycemic and anti-obesity properties in mutant mice with single gene obesity mutations (diabetes, db; obese, ob; viable yellow, Avy). The therapeutic effects differed depending on the mutation as well as the inbred background on which the mutation was maintained. These steroids prevented onset of hyperglycemia and reduced the rate of weight gain in C57BL/6J-db/db and ob/ob mice, whereas in C57BL/KsJ-db/db mice, only hyperglycemia was prevented. The viable yellow (Avy) mutant, exhibiting a more slowly developing obesity condition, responded to all steroids with a marked decrease in rate of weight gain associated with decreased plasma insulin concentrations. Steroid treatment of most mouse mutants was associated with normal or increased food intake, a feature that suggests a decrease in metabolic efficiency. In order to assess any potential energy wastage by steroid stimulation of futile cycles we looked at the rates of lipogenesis, gluconeogenesis and oxygen consumption in steroid-treated normal and mutant mice. With the possible exception of the rate of gluconeogenesis that in obesity mutants was consistently reduced to normal by treatment, no metabolic changes were of sufficient magnitude to account for the marked decrease in metabolic efficiency. All treatments potentiated the action of insulin. This potentiation may change the hormonal balance such that minor changes in the rates of many metabolic pathways may interact to produce a large decrease in metabolic efficiency. PMID- 2970094 TI - Changes in aesthetic appearance and intelligibility of speech after partial glossectomy in patients with Down syndrome. AB - Eight patients with Down syndrome, aged 9 years and 10 months to 25 years and 4 months, underwent partial glossectomy. Preoperative and postoperative videotaped samples of spoken words and connected speech were randomized and rated by two groups of listeners, only one of which knew of the surgery. Aesthetic appearance of speech or visual acceptability of the patient while speaking was judged from visual information only. Judgments of speech intelligibility were made from the auditory portion of the videotapes. Acceptability and intelligibility also were judged together during audiovisual presentation. Statistical analysis revealed that speech was significantly more acceptable aesthetically after surgery. No significant difference was found in speech intelligibility preoperatively and postoperatively. Ratings did not differ significantly depending on whether the rater knew of the surgery. Analysis of results obtained in various presentation modes revealed that the aesthetics of speech did not significantly affect judgment of intelligibility. Conversely, speech acceptability was greater in the presence of higher levels of intelligibility. PMID- 2970095 TI - The peripheral nerve allograft: a dose-response curve in the rat immunosuppressed with cyclosporin A. AB - The potential use of peripheral nerve allografts would significantly improve the reconstructive potential for patients with major peripheral nerve deficits. This study evaluated the response of the nerve allograft recipient treated with varying dosages of cyclosporin A (CsA) to determine the minimal effective dosage necessary to prevent nerve graft rejection. Lewis rats (RT1l) were the recipients of syngeneic nerve grafts from identical Lewis donors or allogeneic nerve grafts from ACI (RT1a) donors. Nerve grafts were inlaid next to the intact sciatic nerve of the recipient. The immunologic responsiveness of the recipient animal's lymphocytes to a donor-specific antigenic challenge was assessed by the mixed lymphocyte reaction (MLR). In addition, nerve grafts were evaluated histologically. Animals were monitored for cyclosporin A toxicity. It was found that cyclosporin A (5 mg/kg per day) was effective in rendering the recipient animals unresponsive by mixed lymphocyte reaction at 10, 20, and 40 days after engraftment. This dosage was similarly effective in preventing histologic changes characteristic of nerve allograft rejection. This dosage regimen was nontoxic to the animals. Our study ascertained a minimal nontoxic dosage of cyclosporin A that effectively prevented nerve allograft rejection across a major histocompatibility disparity in rats. PMID- 2970097 TI - Subclavian steal syndrome and its management by angioplasty. PMID- 2970096 TI - Use and sources of payment for health and community services for children with impaired mobility. AB - A survey was made of the parents of 380 children whose mobility impairments require the use of a wheelchair, walker, or braces. They were asked about equipment, health services, related services, and family support services used during the previous year. There was extensive use of equipment and traditional medical and health services, such as visits to primary care and specialist physicians; there was moderate use of related health services, such as physical or occupational therapy and child counseling; and there was very little use of community-based family support services, such as respite care, after-school care, homemaker services, and summer camp. The cost of health care, particularly medical specialty care, was defrayed in large part by private insurance and public programs, such as Medicaid and Title V Programs for children with special health care needs, while financial support for related services, such as physical therapy and speech therapy, came largely through the schools. Compared to funding for health and related services, financial aid for community-based family support services is largely lacking. PMID- 2970098 TI - Intraluminal stents in atherosclerotic iliac artery stenosis: preliminary report of a multicenter study. AB - Fifteen patients with symptomatic iliac artery stenosis were treated with intraluminal placement of balloon-expandable stents. Before treatment, 14 patients had intermittent claudication, and one had a limb at risk for amputation. One patient had diabetes mellitus, nine had hypertension, and all were long-term smokers. Two patients had surgical placement of the stent; in one patient this was part of a combined revascularization procedure. All other stents were placed percutaneously. The transstenotic gradient after injection of vasodilating drugs distal to the lesion decreased from a mean of 32.3 mm Hg +/- 16.7 to 3.1 mm Hg +/- 4.2 after stent placement. Ankle-arm Doppler systolic pressure index increased from a mean of 0.68 +/- 0.22 to 0.96 +/- 0.24 after the procedure. The treatment eliminated intermittent claudication in 14 patients and increased exercise tolerance to 500 m in the patient with a limb at risk for amputation before the procedure. The improved condition persisted in all patients during the follow-up of 6-12 months. Stent placement may be a valuable adjunct in the management of iliac artery disease. PMID- 2970099 TI - Percutaneous transluminal laser angioplasty with contact probes. AB - Percutaneous transluminal laser angioplasty (PTLA) with a metal contact probe and a sapphire contact probe was performed under experimental conditions and in 88 patients with femoropopliteal artery occlusions. The experiment revealed that more ablation was caused by the sapphire probe than by the metal probe. Because of heat accumulation in subsurface tissue layers the metal probe caused a three to four times larger zone of thermal necrosis surrounding the tissue defect. Clinically both contact probes revealed similar results. The initial rate of recanalization of femoropopliteal artery occlusions with a mean length of 8 cm was 78% (metal probe) to 82% (sapphire probe), and the perforation rate was 14% (metal probe) to 8% (sapphire probe). Doppler ultrasound examinations after 6 months revealed a patency rate of 84%. PTLA has proved to be a successful and safe procedure for recanalization of arterial obstructions that are not appropriate for fibrinolysis. PMID- 2970100 TI - The Palmaz stent: a possible technique for prevention of postangioplasty restenosis. PMID- 2970101 TI - Living conditions of an adult population with Down's syndrome. AB - The living conditions of 38 patients with Down's Syndrome in the Danish county of Aarhus were investigated. All patients were born January 1, 1934 to December 31, 1943. The mean age on July 1, 1984 was 45.7 years, 23 patients (60.5%) were male, 15 (39.5%) female. On census day, December 31, 1984, 82% lived in institutions, 13% in private homes (parents/siblings), and 5% in sheltered residences with other mentally handicapped patients. Out of the 31 institutionalized patients, 21 (68%) lived in old-style institutions. The rest lived in hostels or nursing homes. During part of or the whole year of 1984, 23 (61%) patients worked in sheltered workshops. Seven (18%) had no daily occupation during 1984, and the rest had some sort of occupation. Twenty-seven patients (71%) attended evening classes during 1984, and 36 patients (95%) attended some type of recreation or entertainment in the same year. More than half of the patients got outside their permanent residence at least twice a week (work excluded). During 1984, 27 out of 33 patients living in institutions or in sheltered residences got an average of 15.3 visits, and an averaged 16.7 phone-calls were made to the staff to hear about the well being of 15 out of the 33 patients. PMID- 2970102 TI - Training social skills to severely mentally retarded multiply handicapped adolescents. AB - Three severely mentally retarded, multiply handicapped, adolescents were treated in a classroom setting for social skills deficits. Two of these children exhibited symptoms of autism including periods of echolalia, and fascination with tactile and visual stimulation. One of the pair was deaf. The third child was profoundly mentally retarded and had minimal expressive language skills. All had received sign language training to facilitate communication. Treatment focused on increasing the frequency of eye contact, in seat and response to verbal prompt behaviors, skills deemed necessary to facilitate use of sign language communication and to increase social interaction. Baseline and treatment were evaluated in a multiple baseline, alternating treatment design across children. Baseline was taken on responses to 10 standard questions, asked by the teacher, based on verbal presentation and sign language. This same procedure was then continued during the initial treatment phase following training sessions. During training, the children received social reinforcement, performance feedback and edible reinforcement, in the form of candy, for appropriate performance. Physical and verbal prompts as well as pictorial cues were employed to shape appropriate behavior. In the second treatment phase, training was implemented in the classroom in which baseline data had been collected. Improvement in target behaviors, via training sessions held four days a week, was noted. These data suggest that use of a combination of visual stimuli, operant and social learning methods can remediate social skills deficits in children with multiple psychological and physical deficits. The implications of these findings for current and future research are discussed. PMID- 2970103 TI - [Nutritional status of handicapped children]. PMID- 2970104 TI - [Spontaneous hematoma of the sheath of the rectus muscle in a patient treated with cefotaxime]. PMID- 2970105 TI - Utilizing role theory to assist the family with sudden disability. PMID- 2970106 TI - [The professional field of the psychologist in vocational training--results of a survey]. AB - A questionnaire inquiry was carried out with the Psychological Services of the Vocational Training Centres in the Federal Republic of Germany, directed at their fields of work and their position within the facility. The Psychological Services invariably are involved in the admission procedure, and counselling as well as therapeutic work, individual and in groups, have become a firmly established focus alongside their traditional diagnostic tasks. Aspects of supervision are increasingly being used as new methods of personnel development. Though staffing has improved since 1977, great variations still exist between the various centres, and our findings clearly point out the gaps in service provision related with lower client/psychologist ratios. Following introduction and consolidation of the Psychological Services, a phase of re-orientation seems to be under way, with special attention to focus on interactional processes within the facilities, on implementation of rehabilitation objectives, and on entry of clients with chronic mental impairment. PMID- 2970107 TI - Carotid body chemoreceptor and ventilatory responses to sustained hypoxia and hypercapnia in the cat. AB - To understand the role of carotid chemoreceptor activity in the ventilatory responses to sustained hypoxia (30 min) the following measurements were made in cats anesthetized with alpha-chloralose: (1) carotid chemoreceptor and ventilatory responses to isocapnic hypoxia and to hypercapnia during hyperoxia; (2) carotid chemoreceptor responses to isocapnic hypoxia after dopamine receptor blockade; and (3) ventilatory responses to hypoxia after bilateral section of carotid sinus nerves (CSN). Transition to hypoxia (PaO2 approximately equal to 52 Torr) from hyperoxia gradually increased carotid chemoreceptor activity by ten fold and ventilation by two fold without any detectable overshoot. Termination of isocapnic hypoxia with hyperoxia (PaO2 greater than 300 Torr) at 30 min promptly restored the carotid chemoreceptor activity to prehypoxic level. Ventilation also decreased promptly, but remained above the control value. Induction of hypercapnia (from 31.8 Torr to 43.9 Torr) during hyperoxia was followed by a prompt increase in the chemoreceptor activity by four fold which subsequently diminished, and by a gradual four fold increase in ventilation. Termination of hypercapnia after 30 min was followed by a prompt return of chemoreceptor activity and by a slow return of ventilation to near control levels. Dopamine receptor blockade increased carotid chemoreceptor responsiveness to acute hypoxia but did not alter the response pattern during sustained hypoxia. After bilateral CSN section, ventilation decreased during maintained hypoxia. Thus, a stimulatory peripheral and inhibitory central effects of hypoxia could produce a biphasic ventilatory response to short-term hypoxia in the anesthetized cat with intact CSN but did not manifest it. The results suggest that the chemosensory input not only promptly stimulates ventilation but also prevents the subsequent depressant effect of hypoxia on the brain-stem respiratory mechanisms and hence presumably a biphasic ventilatory response in the anesthetized cat. PMID- 2970108 TI - [Peripheral T-lymphocyte subpopulations in hepatic diseases of alcoholic origin]. PMID- 2970109 TI - [An evaluation of sterility today]. AB - After a reminder of the epidemiology: fertilization ability, fertility, hypofertility, the classic work-up of a sterile couple will be presented: temperature, hormonal evaluation, sperm count, cervical secretions, PCT, hysterogram. With each examination, a critical study of the evaluation of the results will be developed and possibly new ideas will be approached. Such a review reveals, most of the time, the etiology of the sterility. If this is not the case, after systematic laparoscopy, the rate of unexplained sterility will be approximately 5 p. cent. This small number of "unexplained sterilities" could then benefit from the most sophisticated new techniques, especially in-vitro fertilization of which the results, in terms of pregnancy, are identical in tubal pregnancies and unexplained pregnancies. PMID- 2970110 TI - [Laparoscopic control in the treatment of salpingitis. Apropos of 40 cases]. AB - In studying 40 cases of salpingitis diagnosed by laparoscopy, the course of which was controlled with a second laparoscopy, the authors try to evaluate the overall effectiveness of the treatment, the effectiveness according to the type of salpingitis, and finally the effectiveness according to the nature of the antibiotic treatment. PMID- 2970111 TI - Radiographic mobility of the lumbar spine and its relation to clinical back motion. AB - A sample of 194 examinees (117 women and 77 men), representative of the population, with a history of low back pain, were examined clinically and radiographed. The angles between vertebrae at the levels of L4-L5 and L5-S1 were measured from the lateral lumbar erect, maximal flexion, and maximal extension views. Average mobility at L4-L5 was 14.5 degrees in women and 13.4 in men, and at L5-S1 11.5 degrees in women and 12.1 in men. Mobility became more restricted with increasing age both in women and men; especially at L4-L5. In women decreased radiographic mobility at L4-L5 was significantly (p = 0.002) correlated with restricted side-bending and rotation found at the physical examination. These correlations were clearly less significant among men. At L5-S1, restricted extension in women but restricted flexion and Schober's test in men were significantly correlated with decreased radiographic mobility. PMID- 2970112 TI - Low-back pain: pain description as a diagnostic aid. AB - Chronic low-back pain sufferers often present a mixture of somatic and psychologic symptoms, complicating the determination of diagnoses. Two hundred consecutively referred patients with chronic low-back pain had clinical, radiological and laboratory tests performed. The patients were classified into one of the three different categories: a somatic (I) and a somatoform group (III), and an intermediate variety (II). All patients completed a pain questionnaire, giving information about distribution, intensity, variation and somato-sensory qualities of pain in various parts of the body. Patients in group I compared to group III showed differences in pain localization, dispersion and intensity, as well as pain variation and relief. Somatoform patients were mostly females in their forties, their symptoms had started earlier and lasted longer as compared to patients of the intermediate and somatic variety. In diagnosing and dealing with low-back pain patients general practitioners and specialists are recommended to use a simplified pain questionnaire as an aid in sorting the complainers. PMID- 2970113 TI - Assessment of myocardial perfusion with laser Doppler flowmetry. An experimental study on porcine heart. AB - Laser Doppler flowmetry was applied to the arrested heart of four pigs and to the fibrillating heart of three pigs during cardiopulmonary bypass. The coronary blood flow was maintained during cardiac arrest by infusion of hyperkalemic blood into the aortic root. A significant correlation (r = 0.88, n = 52, p less than 0.001) between laser Doppler signal and coronary blood flow was found during cardiac arrest. During ventricular fibrillation after release of the aortic cross clamp there was significant correlation (0.84, n = 38, p less than 0.01) between laser Doppler signal and extracorporeal blood flow. A residual laser Doppler signal of about 60% of the maximal value was recorded even after the bypass flow was discontinued. Laser Doppler flowmetry is concluded to permit measurement of myocardial perfusion in the arrested porcine heart. Muscular activity of the heart contributes to the output signal during ventricular fibrillation. PMID- 2970114 TI - Cutaneous microcirculation and blood rheology following cardiopulmonary bypass. Laser Doppler flowmetric and blood cell rheologic studies. AB - In 23 patients undergoing coronary artery bypass grafting, measurements of cutaneous blood flow were made with laser doppler flowmetry. Simultaneously blood was sampled for measurement of red cell filtration rate (RFR) and plasma-white cell filtration rate (P-WFR). The cutaneous blood flow showed significant overall reduction postoperatively. When the saphenous vein or internal mammary artery was used as bypass graft, the reduction in skin blood flow at the sites from which the vessels were taken was significantly greater than in contralateral, undisturbed sites. RFR and P-WFR were also significantly reduced postoperatively, and these changes showed significant concomitance with the fall in laser doppler flow (LDF%). On postoperative day 6 there was some improvement in LDF% and RFR but further slight deterioration in P-WFR. The study indicated that surgical trauma locally reduces cutaneous blood flow and that trauma to blood cells following cardiopulmonary bypass can contribute to this reduction. PMID- 2970115 TI - Open cardiac operations in patients with abnormalities of white blood cell number or function. AB - Abnormality of white blood cell number or function has been viewed by some as a relative contraindication to operations done with cardiopulmonary bypass. We have operated upon three patients, each with a different white blood cell abnormality, with good results. We do not believe these patients should be denied operation. PMID- 2970116 TI - Delta hepatitis: first two cases identified in Kansas City. AB - Hepatitis delta virus (HDV) was identified in 1977 and has since been found to be a worldwide disease. Prevalence in North America is highest among drug abusers. We present two IV drug abusers from the city hospital in Kansas City as the first two cases identified in the city, one with apparent superinfection and the other with a chronic HDV infection. We review the various manifestations and diagnosis of HDV, and advocate prevention of spread of HDV infection by hepatitis B vaccination and use of single donor blood products in high-risk patients. PMID- 2970117 TI - Traumatic rupture of the thoracic aorta. AB - Between 1968 and 1981, 25 patients with acute traumatic rupture of the aorta were treated at the University of Virginia. Twenty-two of these patients (88%) had serious concomitant injuries. The aortic tear was just distal to the left subclavian artery in 19 patients (76%), and at other sites in six patients (24%). Lacerations were at multiple sites in three patients (12%), in the ascending aorta in one (4%), in the distal aortic arch in two (8%), and in the descending aorta well beyond the subclavian artery in six (24%). Two of the patients (8%) died of free rupture of the aorta before reaching the operating room. The other 23 patients (92%) had operation and 17 (68%) survived. At least ten of the 17 survivors (59%), with an average follow-up of seven years, do not have a disability as a result of the injury. In this group, 24 of the 25 traumatic aortic injuries (96%) occurred in the distal aortic arch or the descending aorta and could be repaired through a left posterolateral thoracotomy. PMID- 2970118 TI - [Organization of medical services for agricultural workers]. PMID- 2970119 TI - [Socio-hygienic aspects of medical care for the disabled]. PMID- 2970120 TI - [Possibilities of improving preventive oncological examinations]. PMID- 2970121 TI - [A hematologist's experience with consultation work in a polyclinic]. PMID- 2970122 TI - Artificial intelligence in the diagnosis of low-back pain and sciatica. AB - In a prospective trial of 200 patients with low-back pain or sciatica, the diagnostic performance of a computer was compared with that of a clinician in a variety of clinical settings. The results indicate that artificial intelligence techniques can be used for the differential diagnosis of low-back disorders, can outperform clinicians, and can be used to develop better methods of human differential diagnosis. PMID- 2970123 TI - Pain drawings in chronic back pain. AB - Pain drawings were obtained from two groups of patients and one of nonpatients, in a total of 264 subjects, all suffering from back pain. The pain drawings were rated in four grades according to the degree of nonorganic and extended pain. The reliability was excellent with an intra- and interrater agreement of 80 and 70%, respectively. Three quarters of the nonpatient group had dull aching pain in the lower back only, whereas widespread or nonanatomical pain was prevalent in patients responding poorly to treatment. A correlation was also found to ethnic background and social situation but not to alcohol abuse or psychiatric illness. Pain drawings afford an important clue to nonorganic factors in the assessment of back pain. PMID- 2970124 TI - Contact dermatitis and related dermatoses associated with petroleum recovery and use. AB - The author reviews the skin's structural and functional protections, and causal factors and clinical patterns of occupational skin disease. He then examines the literature concerning petroleum industry operations and petroleum-derived product use as they relate to skin disease. The chapter concludes with commentary on prevention and treatment of related skin disease. PMID- 2970125 TI - Dose related effects of salbutamol and ipratropium bromide on airway calibre and reactivity in subjects with asthma. AB - The relationship between change in airway calibre and change in airway reactivity after administration of bronchodilator drugs has been investigated by comparing the effect of increasing doses of inhaled salbutamol and ipratropium bromide on the forced expiratory volume in one second (FEV1), specific airways conductance (sGaw), and the dose of histamine causing a 20% fall in FEV1 (PD20) in six subjects with mild asthma. On each of 10 occasions measurements were made of baseline FEV1, sGaw, and PD20 after 15 minutes' rest, and followed one hour later, when the FEV1 had returned to baseline, by a single nebulised dose of salbutamol (placebo, 5, 30, 200 and 1000 micrograms) or ipratropium (placebo, 5, 30, 200 and 1000 micrograms) given in random order. Measurements of FEV1, sGaw, and PD20 were repeated 15 minutes after salbutamol and 40 minutes after ipratropium. Salbutamol and ipratropium caused a similar dose related increase in FEV1 and sGaw, with a mean increase after the highest doses of 0.76 and 0.69 litres for FEV1 and 1.15 and 0.96 s-1 kPa-1 for sGaw. Salbutamol also caused a dose related increase in PD20 to a maximum of 2.87 (95% confidence interval 2.18 3.55) doubling doses of histamine after the 1000 micrograms dose, but ipratropium bromide caused no significant change in PD20 (maximum increase 0.24 doubling doses, 95% confidence interval -0.73 to 1.22). Thus bronchodilatation after salbutamol was associated with a significantly greater change in airway reactivity than a similar amount of bronchodilatation after ipratropium bromide. This study shows that the relation between change in airway reactivity and bronchodilatation is different for two drugs with different mechanisms of action, suggesting that change in airway calibre is not a major determinant of change in airway reactivity with bronchodilator drugs. PMID- 2970126 TI - [Anuria, hepatocellular insufficiency and bone marrow aplasia after the administration of streptozocin and fluorouracil]. PMID- 2970127 TI - [History and society: the plague]. PMID- 2970128 TI - [The publications of Samuel Hahnemann]. PMID- 2970130 TI - [Pediatrics in the 18th century]. PMID- 2970129 TI - Evliya Chelebi's description of medicine in seventeenth-century Egypt. PMID- 2970131 TI - [Pohl for Windaus: "On the optical detection of a vitamin"]. PMID- 2970133 TI - Cell-mediated immunity in the cornea. AB - An alternative model for corneal allografting termed the reverse corneal allograft reaction (RCAR) was developed in this study. Spleen cells from an alloimmunized donor were injected into the corneal stroma of the immunizing donor strain or were restimulated in mixed lymphocyte culture and then injected into the corneal stroma of the immunizing strain. The reaction began as a circular opaque site that spread and became irregularly shaped during the first 5 days after cell injection. The epithelial surface of the cornea became uneven and epithelial cell erosions were noted. Histological examination revealed that corneal stromal keratocytes at the site of inoculation had undergone degeneration and the injected cells had migrated toward the epithelial-stromal boundary, wherein a disruption of the basement membrane and disintegration of the epithelial cells occurred. Purified spleen cell subsets injected separately did not mediate the reaction. A suspension of T lymphocytes and class II antigen positive macrophagelike cells was required to cause the RCAR. This reaction, which mimics a delayed-type hypersensitivity response, was transient, reaching a peak by day 5 and waning by day 8. This experimental model of the corneal allograft reaction shows promise for the study of cells and mediators of the corneal allograft reaction and can be employed as a reproducible system in which to test drug therapies for the treatment of corneal allograft rejection. PMID- 2970132 TI - Use of anti-L3T4 and anti-Ia treatments for prolongation of xenogeneic islet transplants. AB - The effects of T helper lymphocyte and Ia+ cell depletion were examined for their ability to independently and synergistically achieve prolongation of xenogeneic (rat-to-mouse) islet transplants. Recipient mice were depleted of T helper lymphocytes by short-term treatment with the anti-L3T4 monoclonal antibody GK1.5. Donor rat islets were treated prior to transplantation with a concentration of anti-Ia immunotoxin (13.4 x RT) that selectively depleted Ia+ cells within the islets while leaving functional insulin-secreting beta-cells unaffected. Anti L3T4 treatment alone allowed transplants to be prolonged compared with untreated controls; however, all such treated mice rejected their xenogeneic transplant within 22 days. Although 13.4 x RT treatment of donor islets alone did not prolong engraftment, when donor rat islets were pretreated with the anti-Ia immunotoxin and grafted into L3T4-depleted mice, normoglycemia was maintained for greater than 50 days in 56% of transplants. These results suggest that neither L3T4 depletion nor anti-Ia immunotoxin treatment alone is enough to achieve indefinite survival of xenogeneic islets. However, decreasing the immunogenicity of the transplanted islets by anti-Ia immunotoxin treatment prior to transplantation into anti-L3T4 treated mice can allow greatly prolonged xenogeneic graft survival. PMID- 2970134 TI - Immunological tolerance of sheep to skin allografts. AB - A majority of grafts of adult sheep skin placed on fetal lambs at 55 days gestation were retained and increased 50-100-fold in area by the time of birth. Although the original grafts placed in utero survived, second and third grafts from the same donors were rejected. Lambs bearing skin grafts attained full mixed leukocyte reactivity against cells from the skin donors. PMID- 2970135 TI - Expression of HLA antigens on renal tubular cells in culture. II. Effect of increased HLA antigen expression on tubular cell stimulation of lymphocyte activation and on their vulnerability to cell-mediated lysis. AB - Episodes of renal allograft rejection are characterized by an infiltrate of mononuclear leukocytes into the graft and increased HLA antigen expression by graft tubular cells. As HLA antigens are important immune-recognition molecules, we examined whether their increased expression during rejection might contribute to the rejection process. Interferon gamma (IFN-gamma)-treatment of cultured human kidney (HK) cells induced them to increase HLA antigen expression and caused a slight, but nonsignificant increase in their capacity to stimulate proliferation of allogeneic lymphocytes in primary mixed lymphocyte kidney culture (MLKC) (maximum of 8110 +/- 5015 vs. 3966 +/- 4050 counts/min on day 8), which was further increased by addition of IL-1. This proliferation never approached that induced by peripheral blood mononuclear stimulator cells (maximum of 40,325 +/- 10,694 counts/min on day 5), and addition of HK cells to mixed lymphocyte culture inhibited proliferation. There was no difference in lysis of IFN-gamma-treated or untreated HK-cell targets by "specific" cytotoxic effector cells produced in mixed lymphocyte culture using stimulator lymphocytes from the kidney cell donor (49.4 +/- 20% vs. 50.4 +/- 26% specific release in CML). Lysis by 3rd-party cytotoxic effectors produced in MLC using stimulator lymphocytes unrelated to the kidney-cell donor was greater for untreated HK cells (27.4 +/- 20%) than for IFN-gamma-treated HK targets (7.6 +/- 6%, P less than 0.001). IFN gamma-activated naive mononuclear leukocytes lysed untreated HK targets but not IFN-gamma-pretreated targets, and this nonspecific cytotoxicity was mediated by lymphocyte- but not monocyte-enriched cell populations. HK cells are therefore poor stimulators of alloproliferation even when they express increased HLA antigen. They are lysed by both specific and nonspecific effector cells, and exposure to IFN-gamma makes them less vulnerable to nonspecific cytotoxicity and by inference, more vulnerable to specific cytotoxicity. PMID- 2970136 TI - Cyclosphorine sensitivity in renal transplant patients. Quantitative assessment of interleukin-2-producing alloreactive helper T cells by limiting dilution analysis. PMID- 2970137 TI - Synergy between subtherapeutic doses of cyclosporine and immunobiological manipulations in rat heart graft recipients. AB - Cyclosporine in combination with other chemical or biological immunosuppressive modalities has been useful in clinical and experimental organ transplantation. In these studies, the efficacy of adjunctive subtherapeutic doses of CsA given to immunologically enhanced heart graft recipients or to animals treated with an anti-IL-2 receptor monoclonal antibody (ART18) are described. Individually, the treatment entities are only partially effective. In rats undergoing active and passive enhancement alone, heart allograft survival was increased to 25 +/- 12 days in two-thirds, indefinitely in one-third. After ART18 treatment, grafts survive 21 +/- 1 days. Grafts are accepted permanently in animals receiving full dose CsA (15 mg/kg X 7), but are rejected acutely (c. 7 days) when subtherapeutic doses (1.5 mg/kg X 7) are used. However, when subtherapeutic doses of CsA are given in combination with immunological enhancement or with interleukin-2 receptor-targeted therapy, graft survival increases dramatically, with permanent or markedly prolonged engraftment occurring in all instances. In the early phases of host unresponsiveness, both enhancement and IL-2R-targeted therapy, graft survival increases dramatically, with permanent or markedly prolonged engraftment occurring in all instances. In the early phases of host unresponsiveness, both enhancement and IL-2R-targeted therapy spare selectively T cells with suppressor activity in vivo; in enhanced animals, the W3/25+ subset is responsible for prolonged graft survival, the OX8+ fraction is responsible in ART18-treated animals and in CsA-treated animals. Both subpopulations show suppressor activity in the later stages of combination treatment. IL-3 production is increased significantly in these states of unresponsiveness, an observation also noted during maintenance CsA treatment; this seems to correlate with suppressor activity. Immunoperoxidase studies of the graft infiltrates emphasize the synergistic effects of combination treatments. Thus, subtherapeutic doses of CsA plus biologic host manipulations produce greatly increased graft survival by affecting selectively different host immune mechanisms. PMID- 2970138 TI - Mechanisms of suppression in mixed allogeneic chimeras. AB - Cells with the ability to suppress cytotoxic T lymphocyte generation are found in the spleens of whole-body-irradiated (WBI) mixed allogeneic and syngeneic bone marrow transplant recipients in the early weeks after BMT. Previous studies have indicated that suppression is mediated by "null cells" similar to natural suppressor (NS) cells (1), and have ruled out several possible trivial explanations for the suppressive effect. We report here the results of additional experiments designed to assess possible mechanisms of suppression. We compared the cell populations after 5 days' incubation of cultures containing normal responding splenocytes plus irradiated allogeneic stimulator cells, with or without a cocultured suppressive chimeric splenocyte population. The data indicate that total viable cell yields are only slightly reduced, if at all, in suppressed cultures, but that the proportion of T cells is markedly reduced as measured at the end of the incubation period. Splenocytes from early BMT recipients do not appear to proliferate during the suppression of a mixed lymphocyte culture, and such populations represent only 15% of cells at the end of the 5-day incubation period. Suppression is strongest when the suppressive population is added at the initiation of MLC, and is lost if addition is delayed beyond day 3. Suppression can be overcome by T cell growth factor (TCGF)--and, to a lesser extent, by recombinant IL-2 (rIL-2), although resting suppressive populations do not consume appreciable amounts of these lymphokines. These results therefore suggest that suppression in MLC may occur primarily during the induction of helper T lymphocytes. PMID- 2970139 TI - Molecular characteristics of cyclophilin-cyclosporine interaction. AB - The ability of cyclophilin to react with derivatives of cyclosporine (CsA) was studied. Cyclophilin was found to interact preferentially with CsA-residues 1, 2, 10 and 11, which, together with residue 3, are the residues known to contribute to the immunosuppressive activity of CsA. The recognition of different CsA derivatives by cyclophilin was correlated with their immunosuppressive activity in vitro. All CsA-derivatives showing a significant activity did bind to cyclophilin, although some of the CsA-derivatives able to bind cyclophilin exhibited only low activities. The results suggest that binding to cyclophilin might be one requirement for immunosuppressive activity of CsA derivatives. When tested with CsA-derivatives showing various conformational changes, the binding of cyclophilin was strongly specific for the peptide-ring conformation of CsA. No binding of calmodulin to CsA could be detected in several formats of solid-phase enzyme- or radioimmunoassay, suggesting that, in contrast to cyclophilin, calmodulin does not possess sufficient affinity for CsA to bind to it when immobilized on the solid phase. PMID- 2970141 TI - Frequency of human alloantigen-reactive T lymphocytes. III. Evidence that cyclosporine has an inhibitory effect on human CTL and CTL precursors, independent of CsA-mediated helper T cell dysfunction. AB - We have used limiting dilution analysis to study the behavior of alloantigen reactive cytolytic T lymphocytes derived from human peripheral blood. During these studies, we found that the presence of cyclosporine in limiting dilution microcultures significantly impairs the subsequent development of alloantigen reactive cytolytic T cell activity. As a result, CsA reduces the estimate of CTL precursor frequency by limiting dilution analysis. CTL frequency estimates are reduced by CsA in a dose-dependent manner, and concentrations of CsA that are readily achieved in human peripheral blood (100-1000 ng/ml) are capable of reducing estimates of CTL frequency by 90% to 100%. Further studies revealed that (1) human CTL derived either from fresh peripheral blood or from primary mixed lymphocyte cultures are sensitive to the suppressive effects of cyclosporine in limiting dilution microcultures, indicating that CsA influences both alloantigen primed CTL and CTL precursors; (2) CsA impairs an immunologic event or events, that occurs for at least the first four days of limiting dilution microculture incubation; (3) CsA-mediated suppression is eliminated by separation of CTL from cyclosporine; (4) CsA blocks development of CTL generation, but not cell proliferation in limiting dilution microcultures; and (5) the CsA-mediated suppression is not reversed by supraoptimal concentrations of IL-2, high concentrations of gamma-IFN, or supplementation with the multiple lymphokines present in MLC supernatants. These data suggest that CsA may have a direct inhibitory influence on the differentiation of human CTL precursors that is independent of helper T cell dysfunction. PMID- 2970140 TI - Inverse correlation of cyclosporine binding with sensitivity and resistance. AB - The primary effects of CsA on human lymphocyte responses in vitro appear to be the inhibition of IL-2 production and the inhibition of cytotoxic T cell activation. Induction of suppressor T cell activity is resistant to the effects of CsA. These data imply two distinct subsets of lymphocytes: CsA-resistant and CsA-sensitive. The current studies used a bioactive, dansylated derivative of CsA (dans-CsA), which is fluorescent, to assess binding of CsA at the single-cell level by flow cytometry. The results demonstrate that two populations of cells can be distinguished based on differential staining with dans CsA--a weakly staining subset and a population that binds intensely. Both subsets consist of CD4 (helper) and CD8 (cytotoxic/suppressor) T lymphocytes. Functional analysis revealed that the weakly staining subset consists of IL-2-producing T cells and precursor cytotoxic T lymphocytes. On the other hand, the intensely staining subset includes T cells that suppress in an antigen-specific manner after activation with alloantigen. Further studies showed that the weakly staining subset is markedly sensitive to the immunosuppressive effects of CsA in a PHA stimulation assay, while the intensely binding population is markedly resistant, requiring 10- to 100-fold more CsA to inhibit the PHA response. These studies suggest that sensitivity and resistance to CsA is inversely correlated with binding. PMID- 2970142 TI - Fine-needle aspiration cytology during treatment with orthoclone monoclonal antibody OKT3 for acute cellular rejection after renal transplantation. PMID- 2970143 TI - Cytotoxic cells may not be responsible for renal allograft rejection. PMID- 2970145 TI - Magnetic resonance imaging of the retroperitoneum. AB - Magnetic resonance imaging (MRI) represents the most recent development in imaging of the retroperitoneum. This article discusses the utility of MRI in evaluating the abdominal aorta, inferior vena cava, lymph nodes, and psoas muscle. PMID- 2970144 TI - Angle independent ultrasonic blood flow detection by frame-to-frame correlation of B-mode images. AB - We have previously reported initial clinical results of a novel blood velocity imaging technique utilizing a two-dimensional correlation search applied to consecutively acquired echoes. In this paper, we describe both the physical principles underlying this technique and test tank experiments which define its performance under a variety of conditions. The results indicate that, unlike Doppler flow imaging systems, this technique defines the flow velocity vector in two dimensions and is not subject to aliasing. PMID- 2970146 TI - [Prevention of the suppuration of laparotomy wounds]. PMID- 2970147 TI - [Laparoscopic microcholecystostomy and its complications]. AB - An original method of laparoscopic microcholecystostomy is proposed based on using a spiral catheter and 3 variants of performing this operation. Causes of complications are analyzed. The method of laparoscopic microcholecystostomy used in 426 patients enabled the authors to reduce the incidence of complications from 11.6% to 3.7%. PMID- 2970148 TI - [Complications of non-clostridial anaerobic peritonitis]. AB - Under observation there were 71 patients with non-clostridial anaerobic peritonitis, 37 of them having complications--suppurations of operation wounds, phlegmons of the abdominal wall, abscesses of the abdominal cavity, postoperative non-clostridial peritonitis. Fourteen patients died (37.8%). PMID- 2970149 TI - [Determining the patency of the distal vascular bed and transplant during operations on the arteries]. AB - The article describes the method of quantitative intraoperative assessment of patency of the distal vascular bed and the transplant based on the imitation of blood flow in the vascular bed located lower to the occlusion level. It can be fulfilled by introduction of the liquid under permanent pressure of 120 mm Hg and measurement of its volume per 1 min. The method was used in 85 patients. PMID- 2970150 TI - Pasture study of two types of oxfendazole pulse release bolus for controlling nematodes in calves. AB - One group of first-season calves was dosed with an oxfendazole pulse release bolus at spring turnout (April 30) and on July 15 a second group received the front-loaded oxfendazole pulse release bolus. The objective was to test the boluses for the prophylaxis or control of nematodiasis. The control group consisted of calves to which no bolus was administered. The three groups occupied separate but adjacent plots. For the first five weeks of the trial, three calves, artificially infected with Dictyocaulus viviparus grazed in each plot. Parasitic bronchitis severely affected the control calves, necessitating repeated emergency treatment, whereas administration of the bolus at turnout almost completely prevented this condition. D viviparus infection increased markedly on the control herbage in July and August but was eliminated by the end of June on pasture grazed by bolus treated calves. Treatment in mid-season with the front-loaded bolus brought an outbreak of parasitic bronchitis under control. Gastrointestinal worm egg output was satisfactorily suppressed after the administration of both boluses, resulting in reduced levels of herbage infection. Calves treated with a bolus at turnout gained significantly more weight than either the controls (P less than 0.001) or the calves treated with a front-loaded bolus in mid-season (P less than 0.01). The weight-gain of the calves treated with a front-loaded bolus was slightly but not significantly greater than that of the control calves. On the basis of faecal egg counts, the first pulse released from the standard boluses was delayed and one front-loaded bolus failed to release a dose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970151 TI - Hares and transmission of dictyocaulus. PMID- 2970152 TI - Differential effect of adenovirus 2 E3/19K glycoprotein on the expression of H 2Kb and H-2Db class I antigens and H-2Kb- and H-2Db-restricted SV40-specific CTL mediated lysis. AB - The E3/19-kDa glycoprotein (E3/19K) coded by adenovirus type 2 (Ad2) is known to inhibit the cell-surface expression of major histocompatibility complex (MHC) class I antigens by binding to the MHC antigens intracellularly, and thus reduces recognition of antigens by MHC-restricted cytotoxic T lymphocytes (CTL). We have studied the effect of the E3/19K expression in SV40-infected monkey cells, TC-7/H 2Kb and TC-7/H-2Db expressing transfected H-2Kb and H-2Db antigens, respectively, on the cell-surface H-2 class I antigens and on lysis of the cells by SV40 large tumor (T)-antigen-specific H-2Kb- and H-2Db-restricted CTL clones. H-2Db antigen expression on TC-7/H-2Db cells was drastically reduced by infection with Ad2 but not with an E3/19K-negative SV40-Ad2 hybrid virus, Ad2+ND1, as early as 12 hr postinfection. However, H-2Kb antigen expression on Ad2-infected TC7/H-2Kb cells remained unaltered, even at 24 hr postinfection. Specific lysis of SV40-infected TC-7/H-2Db cells by H-2Db-restricted SV40 T-antigen-specific CTL clones, Y-1 and Y-3, was strongly reduced by coinfection of the target cells with Ad2 but not with Ad2+ND1. Lysis of SV40-infected TC-7/H-2Kb cells by a H-2Kb-restricted SV40 T-antigen-specific CTL clone Y-4 was also reduced significantly by Ad2 infection, but not Ad2+ND1. These results indicate that the E3/19K protein affects cell surface expression of H-2Db antigen but not H-2Kb antigen. PMID- 2970153 TI - Role of the adenovirus 72-kDa DNA binding protein in the rapid decay of early viral mRNA. AB - Previous experiments have shown that the early adenovirus E1A and E1B mRNAs decay with a half-life of 20 min in a lytic infection dependent on the action of the viral 72-kDa DNA binding protein. In contrast, the same E1A and E1B mRNAs are stable when synthesized in 293 cells, an adenovirus-transformed cell line that is devoid of the 72-kDa protein. If 293 cells are infected with the E1A deletion mutant dl312, the endogenous E1A RNA disappears after 4 hr of infection, a time coincident with the appearance of the 72-kDa protein. The induction of decay is specific since there is no decrease in the level of actin or certain other cellular mRNAs. Thus, the stability of the early RNAs is variable and correlates with the presence of the 72-kDa protein. An interaction of the 72-kDa DNA binding protein with RNA inside the cell has been demonstrated by in vivo crosslinking of protein to RNA. However, the protein is found in association with actin mRNA as well as E1A mRNA. Thus, although the 72-kDa protein appears to be required for the rapid decay of viral mRNA it apparently does not impart specificity to the process. PMID- 2970154 TI - [Pseudomembranous colitis treated with teicoplanin]. PMID- 2970155 TI - [A rare case of abdominal pregnancy]. PMID- 2970156 TI - [Disorders of object-naming in Huntington chorea]. PMID- 2970157 TI - [Kwashiorkor syndrome in exudative enteropathy]. AB - We report on a case of a 45 years old man suffering from ulcerative colitis accompanied by a protein-loosing-syndrome (hypoalbuminaemia). The significant decrease of cholinesterase (CHE) and quick level, edemas of the legs and forearms, pleural effusion and ascites had been misinterpreted as signs of liver insufficiency. Low protein diet was given and, thus, the full picture of kwashiorkor developed soon as a logical consequence. Liver-induced fluid dysbalance was assumed and the low-protein diet continued. We describe the difficulties of treatment and the favourable outcome of the disease. PMID- 2970158 TI - [Combined topical treatment of acne with erythromycin and tretinoin]. AB - Vitamin A acid is the most effective comedolytic agent in the therapy of acne vulgaris. Antibiotics are suitable for the treatment of inflammatory lesions (papulo-pustules). Even topically applied, some antibiotics show a sufficient anti-inflammatory effect. Above all, erythromycin is reliable in the topical treatment of acne. Combined therapy with both topical tretinoin and erythromycin is more effective than either alone. During the first weeks of treatment, tretinoin leads to temporary deterioration of the disease, which can mostly be avoided by the anti-inflammatory effect of erythromycin simultaneously applied. PMID- 2970159 TI - [1 percent clindamycin phosphate solution versus 5 percent benzoyl peroxide gel in papulopustular acne]. AB - In a multicenter study, 60 patients suffering from mainly moderate papulopustular acne were treated to pically either with 1% clindamycin phosphate in alcoholic solution or with 5% benzoyl peroxide gel, 30 patients each. During the treatment period of 9 weeks, clinical controls with lesion counts were performed every 3 weeks and after a 3 weeks follow-up. The therapeutic efficacy according to the reduction of papules and pustules was 72% in the benzoyl peroxide group and 73% in the clindamycin group, the latter showing significantly less side effects. On account of its good therapeutic results, we consider topical treatment with clindamycin phosphate an important alternative to systemic antibiotic or topical benzoyl peroxide therapy in acne. PMID- 2970160 TI - [A fluorescence-activated cell sorter--analysis of peripheral T lymphocyte subpopulations in psoriasis]. AB - By means of the fluorescence activated cell sorter (FACS), we analyzed the T lymphocyte subpopulations in peripheral blood samples from 82 patients showing various clinical manifestations of psoriasis. With regard to both the absolute values of T-helper and T-suppressor cells and those of the T-helper/T-suppressor cell ratios, there were no quantitative differences between the patients (i.e. either the entire collective of classified according to the various forms of psoriasis) and the control group. We discuss the pathophysiological relevance of peripheral and in situ analyses of T-lymphocyte subpopulations. PMID- 2970161 TI - [Allergic contact dermatitis caused by etofenamate]. AB - We report on 3 cases of contact dermatitis following topical application of etofenamat. Each patient developed positive reactions in patch tests with etofenamat in concentrations of both 10% and 1%. After testing, one patient showed secondary inflammatory reactions at the original site of application. Considering the frequent administration of etofenamat, contact sensitization seems to be relatively rare. PMID- 2970162 TI - [Physiologic importance of atrial natriuretic peptide in volume expansion and hypertension: experimental studies]. AB - The release of atrial natriuretic peptides (ANP) is stimulated by acute volume expansion. In order to elucidate the importance of the ANP release for the physiological response to volume expansion, a monoclonal antibody was used in rats. This antibody against ANP specifically blocks the ANP effects in vivo. The acute natriuresis induced by isotonic volume expansion is blocked by the antibody. However, the excretion of an additional sodium load (saline with an additional amount of 50 mmol/l sodium) is not entirely ANP-dependent. Acute volume expansion induces a dramatic decrease of plasma renin activity and a slight decrease of plasma aldosterone concentration measured 5 min after volume expansion. These effects are blocked by simultaneous administration of the monoclonal antibody. The endogenous level of ANP that can be reached in volume expansion is thus sufficient for suppression of the renin-aldosterone system. The physiological importance of ANP underlined in these volume expansion experiments raises the question of its role in pathophysiological states such as hypertension. In old, spontaneously hypertensive rats (SHR) ANP plasma levels and relative heart weights are higher than in age-matched normotensive Wistar Kyoto rats (WKY).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970164 TI - [Methodological problems in the radioimmunologic measurement of ANP (atrial natriuretic peptide)]. AB - Parameters influencing the measurement of hANP are designated and discussed on the basis of data from the literature and from our own results. Plasma hANP is dependent on the anatomical location of blood withdrawal, posture, state of hydration and salt load and medications. Interfering substances contained in plasma, management of samples and the type of assay used are of utmost importance. Direct RIA of hANP appears to be particularly sensitive to interferences. Preextraction of hANP from EDTA plasma by octadecylsilica cartridges, followed by RIA seems to be the most trustworthy method for measurement of hANP. Nevertheless, with this method too, we obtain different values for identical plasma samples if we use different antisera. Additionally, lipemia significantly lowers the ascertainable amount of endogenous, but not of exogenously added alpha-hANP. Therefore, it is reasonable to conclude that in preextracted samples too, hANP is not a homogeneous species. We observe a time dependent increase in plasma hANP, reaching a final stable plateau value (about three times the initial values) after about 1.5 h, if EDTA blood samples are left on ice for prolonged time. Hemolysis interferes significantly with hANP determination. Readings of hANP values are reduced in dependence on plasma haemoglobin concentration. According to our experience, EDTA plasma can be stored for several months in either lyophilized form or deep frozen at -20 degrees C without the risk of reduction of hANP immunoreactivity. The data presented in this paper clearly show, that measurement of hANP is susceptible to faults in several details. Results should therefore be viewed critically. PMID- 2970163 TI - Interrelationship between atrial natriuretic peptide and plasma renin, aldosterone and catecholamines in hepatic cirrhosis: the effect of passive leg rising. AB - To assess the role of atrial natriuretic peptide (ANP) in relation to the sympathoadrenal and renin-angiotensin-aldosterone system, and sodium excretion, 88 cirrhotic patients (mean age 52 years; 28 compensated, 28 decompensated with ascites, and 32 decompensated and treated with diuretics) and 26 control subjects were investigated. Basal ANP levels were not different between any group of cirrhotics and controls. Circulating ANP was not related to elevated plasma noradrenaline and adrenaline, and enhanced plasma renin/aldosterone levels in ascitic patients. Furthermore, ANP was not related to urinary sodium excretion, blood pressure, and heart rate. In 30 cirrhotic patients (12 compensated, 18 decompensated with ascites including eight on diuretic therapy) and nine controls, a passive leg rising procedure for 1 h was performed in order to augment central blood volume and atrial pressure physiologically. Ascitic patients (with and without diuretic treatment) experienced a slight but significant increase in plasma ANP indicating preserved responsiveness of ANP release in cirrhosis. Plasma aldosterone was markedly depressed. The data support the underfilling concept of ascitic formation in advanced stages of cirrhosis. The failure of enhanced ANP release under basal conditions may be due to the diminished effective blood volume, resulting in insufficient atrial stretching. PMID- 2970165 TI - [Atrial natriuretic factor in patients with type 1 diabetes mellitus in various stages of diabetic nephropathy]. AB - In this study we investigated circulating atrial natriuretic factor (ANF) concentrations in patients with different stages of diabetic nephropathy. Type 1 diabetic patients with incipient or overt nephropathy had elevated plasma ANF concentrations when compared with nonnephropathic diabetics and normals. Patients having a hyperfiltration only, which might be a risk factor for the development of diabetic nephropathy, had also elevated ANF concentrations. No correlation could be found between ANF concentrations and blood pressure, diabetes duration, age, or parameters of metabolic control. These findings may indicate a role of ANF in the development of diabetic nephropathy. Long-term follow-up studies should be performed to assess the prognostic and diagnostic importance of plasma ANF determinations in diabetic patients. PMID- 2970166 TI - [What pathophysiologic significance does increased plasma levels of human atrial natriuretic peptide have in patients with diabetes mellitus?]. AB - Hypertension is more frequently found in patients with diabetes mellitus than in subjects with normal glucose tolerance. On the other hand, concomitant hypertension accelerates the progression of diabetic nephropathy. To examine whether human atrial natriuretic peptide (human ANF-[99-126], hANP) is involved into the pathogenesis of hypertension and nephropathy of diabetic patients and to find out whether the detection of increased hANP levels can serve as an early marker, helping to identify diabetic patients at increased risk of developing these diabetes complications, we studied 107 randomly selected patients with Type 1 or Type 2 diabetes mellitus (53 women, 54 men). There were no differences between patients with normal hANP levels and patients with hANP levels above normal range regarding age, diabetes duration, metabolic control, kidney function (creatinine clearance and proteinuria), electrolytes, and in plasma renin activity, aldosterone, epinephrine and norepinephrine levels in plasma. However, higher blood pressure was measured and antihypertensive therapy was found more frequently in patients with increased hANP levels (p less than 0.05). This was confirmed by analyzing the subgroup of patients with normal blood pressure without antihypertensive therapy: Again, diastolic blood pressure was found to be higher (p less than 0.05) in patients with elevated hANP than in patients with normal hANP levels. In this subgroup, increased creatinine clearance tended to be found more frequently among patients with increased hANP levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970167 TI - [Plasma ANP levels and its relation to electrolyte and water regulation in neurosurgical intensive care patients]. AB - We examined the relation between plasma atrial natriuretic peptide (ANP) and the changes of the regulating hormones ADH and renin aldosterone in 20 neurosurgical intensive care patients. All patients suffered from elevated intracranial pressure due to severe head trauma or severe subarachnoidal hemorrhage of the anterior circulation. Under controlled mechanical hyperventilation (CMV) with PEEP, 15 patients without evidence of central dysregulation showed no change in plasma ANP, ADH and aldosterone. In five patients with severe central dysregulation of either electrolytes or blood pressure, increases of plasma ANP of various degrees could be observed together with a decline in serum aldosterone. PMID- 2970168 TI - [Plasma levels of atrial natriuretic peptide (ANP) in patients with paroxysmal supraventricular tachycardia and concomitant polyuria (urina spastica)]. AB - A concomitant diuresis and natriuresis has been reported to occur in 30-50% of patients with paroxysmal supraventricular tachycardia. While the increase of the diuresis may be secondary to an inhibition of vasopressin, the etiology of the natriuresis is presently not well understood. To determine the role of atrial natriuretic peptide (ANP) in the pathogenesis of tachycardia-induced polyuria, we measured ANP levels in two female patients (aged 59 and 68 years) with recurrent episodes of paroxysmal supraventricular tachycardia during such attacks, as well as after conversion to a normal sinus rhythm. In both patients, episodes of supraventricular tachycardia were associated with both a large increase in ANP plasma levels (560 resp. 1.208 pg/ml; normal range: 50 +/- 10 pg/ml) and polyuria. Despite elevated ANP plasma levels, no diuretic and natriuretic response to tachycardia could be observed during episodes of shorter duration, some of which occurred in the older patient studied. After restoration of a normal sinus rhythm, ANP plasma levels decreased to baseline levels (107 resp. 32 pg/ml). The results of this study show that the secretion of ANP is increased during episodes of paroxysmal supraventricular tachycardia, suggesting that ANP might contribute to the pathogenesis of natriuresis and diuresis frequently associated with supraventricular tachycardia. PMID- 2970169 TI - A sensitive radioimmunoassay of atrial natriuretic peptide in human plasma; some guidelines for clinical applications. AB - We report the development of an in-house RIA for human alpha-ANP. The method requires the extraction of EDTA-plasma on C18-cartridges followed by radioimmunoassay of the eluates with an antibody raised in sheep against human alpha-ANP coupled to Keyhole Limped Hemocyanin, and a monoiodotyrosyl-alpha-ANP tracer purified with HPLC before use. The assay is precise and sensitive: intra- and interassay coefficients of variation were 8.6% and 11.6%, respectively, and the lower limit of detection was 0.8 pg/tube (4.0 pg/ml plasma). Normal values were 26.0 +/- 15.5 (mean +/- sd) pg/ml plasma (n = 25). Change from supine to sitting position significantly lowered the ANP immunoreactivity. No episodic variations were observed. Our method was applied in a study comparing plasma ANP values in samples of normal persons and CHF-patients. Assay of ANP after extraction using four different antibodies showed significant correlations. HPLC analysis of both plasma of CHF patients and plasma of normal persons revealed the existence of two major immunoreactive components besides alpha-ANP which were not detected in a radioreceptor assay. PMID- 2970170 TI - Potential pitfalls in the radioimmunoassay of physiological plasma levels of atrial natriuretic peptide. AB - Certain aspects of current radioimmunoassay methodology are presented which may restrict accurate measurement of physiological changes in circulating plasma levels of atrial natriuretic peptide. These include the method of sample collection and time of storage, sample extraction, concentration and reconstitution, the homogeneity of tracer and standards, separation of bound from free ligand and the assessment of blank values. If left unchecked, inaccuracies at each of these stages may cumulatively result in unreliable assessment of changes in circulating ANP levels and limit further understanding of its physiological role. PMID- 2970171 TI - [Standardization of plasma determination of atrial natriuretic peptide (ANP)]. AB - Comparison of normal values for plasma levels of atrial natriuretic peptide revealed a tremendous variability: the highest values being 10-times higher than lower mean values. We studied the impact of age, posture, sodium intake and the conditions of processing on the levels of ANP in normal control subjects. Middle aged healthy subjects had p-ANP levels of 56 +/- 10 pg/ml compared to 25 +/- 10 pg/ml in 20-30-year-old students. Upright posture was followed by a decline in p ANP of 20% whereas renin concentration rose 6-fold during upright posture. P-ANP increased during high sodium intake to 134 +/- 9 pg/ml, compared to 55 +/- 7 pg/ml during low sodium intake. We did not find a significant variability of p ANP in the morning hours. Furthermore, when blood was stored at room temperature for 60 min, ANP levels remained unchanged. These variables should be taken into account when a control range is set up. PMID- 2970172 TI - [Significance of atrial natriuretic peptide in heart failure: experimental findings]. AB - We investigated the role of atrial natriuretic peptide (ANP) in a rat model of chronic heart failure (CHF) (coronary ligation) by four different studies: (1) stimulation of secretion of ANP by volume loading, (2) infusion of 1 micrograms/min of rat ANP (99-126), (3) correlations of pressure measurements of right atrium (RAP) and LVEDP versus plasma ANP levels and ANP mRNS versus infarct size, (4) blocking endogenous plasma levels of ANP with monoclonal ANP antibody. In severe CHF, volume loading did not exert a significant increase in plasma ANP, in contrast to moderate CHF or to normal animals. Infusion of ANP reduced RAP and LVEDP without significant increase in renal blood flow (radioactive microspheres) in rats with severe cardiac failure, in contrast to normal animals. A close relationship was found between LVEDP and atrial plasma levels. After injection of ANP antibody, filling pressures and systemic vascular resistance increased, indicating that removal of ANP may enhance arterial vasoconstriction or elevated endogenous ANP to exert a vasodilatory action in severe heart failure. Nevertheless, the cardiocirculatory effects, primarily on renal blood flow, appear to be limited. PMID- 2970173 TI - [Diagnostic and prognostic value of plasma ANP determination after myocardial infarction]. AB - In the present study the prognostic value of ANP measurement was investigated in 52 patients undergoing coronary angiography. 22 normotensive and 30 hypertensive subjects were included. A significant inverse correlation between left ventricular ejection fraction (EF) and plasma ANP was found in normotensives, but not in hypertensives. In patients with chronic myocardial infarction, there was no difference in ANP levels compared to patients without infarction in either group, provided that EF was normal. However, hypertensives showed a 35% increase in ANP compared to normotensives. This was true for subjects with and without myocardial infarction. These results show that in normotensives ANP levels have a prognostic value on a statistical rather than an individual basis. This does not apply to hypertensives, whose ANP level is increased by factors other than impaired ejection fraction. PMID- 2970174 TI - [Does the measurement of plasma ANP have a diagnostic or prognostic value in patients with organic heart disease?]. AB - Plasma levels of atrial natriuretic peptide (ANP) were measured in patients with organic heart disease undergoing diagnostic cardiac catheterization. Independent of nature and duration of the disease (valvular heart disease, congestive cardiomyopathy) plasma ANP levels were closely related to the severity of cardiac failure. Furthermore, plasma ANP levels were found to be negatively correlated with the cardiac index and to be positively correlated with right and/or left atrial and with pulmonary artery pressures. During physical exercise (bicycle ergometer) a marked increase of plasma ANP levels was observed, which was closely related to increments in mean pulmonary artery pressure. This rise in plasma ANP levels during physical exercise was not attenuated in patients with already elevated resting plasma concentrations of ANP. In patients with congestive cardiomyopathy, afterload-reduction by ACE-inhibition resulted in changes of central hemodynamics, which were closely reflected by venous concentrations of ANP. The measurement of plasma ANP levels may serve as an indicator of the severity of cardiac failure. Plasma concentrations of ANP, however, are neither helpful in establishing the etiology of the underlying heart disease nor in differentiating left and right heart failure. However, in cases of already established organic heart disease plasma ANP levels may be used as a marker for assessing the efficacy of the therapeutic regimen. PMID- 2970175 TI - [Pharmacologic effects of atrial natriuretic peptide in humans]. AB - Since the discovery of atrial natriuretic peptide (ANP) by de Bold in 1981, several scientists (e.g. nephrologists, endocrinologists, cardiologists) have contributed to research efforts to define the physiologic and pharmacologic profile of the peptide. ANP given in pharmacologic doses seems to lower blood pressure, probably by reducing both preload and afterload. ANP increases renal water and electrolyte excretion, although the underlying mechanisms have not been entirely clarified yet. The depressor and natriuretic effect of ANP act in harmony with an inhibitory effect on the release of renin, aldosterone, and arginine-vasopressin. While the physiologic role or importance of ANP is still in question, synthetic alpha ANP has a promising pharmacologic profile in renal and cardiovascular pathologic states. PMID- 2970176 TI - [Clinical relevance of the determination of plasma atrial natriuretic peptide in children with chronic renal failure]. AB - To investigate whether plasma atrial natriuretic peptide (ANP) is a valuable index to detect fluid expansion in children with endstage renal disease, we studied 34 children and adolescents on regular intermittent hemodialysis or on continuous ambulatory peritoneal dialysis (CAPD). In 22 pediatric patients, plasma ANP was markedly elevated prior to hemodialysis and fell to near normal levels after dialysis (109.9 +/- 80.0 to 39.0 +/- 23.1 fmol/ml, means +/- SD). During removal of fluid excess by 1-h sequential ultrafiltration without dialysis in eight adolescents, plasma ANP fell from 123.8 +/- 97.0 to 45.3 +/- 24.6 fmol/ml. Ultrafiltration was followed by 3 h hemodialysis with fluid removal and ANP decreased further to 29.6 +/- 12.1 fmol/ml. In 12 patients on CAPD, plasma ANP was similar to that in 96 healthy control children (29.3 +/- 28.9 vs 23.9 +/- 11.9 fmol/ml) and did not differ from that in 12 children with advanced chronic renal failure (32.6 +/- 20.1 fmol/ml) and seven successfully transplanted children (32.4 +/- 8.5 fmol/ml). Four patients on hemodialysis (post dialysis), one on patients on hemodialysis (post dialysis), one on CAPD and two with advanced chronic renal failure had definitely elevated ANP levels (greater than 56 fmol/ml = greater than 3 SD mean of controls) indicating fluid volume overload. It is concluded that plasma ANP is elevated in children with chronic renal failure due to volume expansion and that measurement of plasma ANP is a sensitive marker to detect fluid overload in these children. PMID- 2970177 TI - [Elevated plasma levels and heterogeneity of human atrial natriuretic peptide in patients with progressive chronic renal failure]. AB - The role of human atrial natriuretic peptide (alpha-hANP) in chronic blood pressure (BP) and extracellular fluid volume (ECFV) regulation remains elusive. Hence, the role of alpha-hANP in chronic renal failure is of particular interest since in this pathological condition: (1) increased sodium and water retention plays a major pathogenetic role in the development of hypertension and (2) altered secretion and/or metabolism of alpha-hANP may contribute to fluid volume and BP regulation. To evaluate the relationship between the degree of renal insufficiency, BP and circulating alpha-hANP, we determined plasma alpha-hANP concentrations by radioimmunoassay in 16 nondialyzed patients with progressive chronic renal failure (CRF) of various degrees; subsequently analysis of potential molecular heterogeneity of immunoreactive (ir) ANP was performed by means of gel permeation of plasma extracts from patients with CRF without concomitant hypertension. Serum creatinine concentrations ranged from 127 to 1187 (435 +/- 76) mumol/l, systolic BP from 135 to 200 (158 +/- 4) and diastolic BP from 80 to 110 (94 +/- 2) mmHg, respectively. Plasma alpha-hANP concentrations ranged from 5 to 75 (23 +/- 4) pmol/l which was thus significantly higher as compared to 9 +/- 2 pmol/l found in healthy volunteers (p less than 0.05). A highly significant linear correlation between plasma alpha-hANP and serum creatinine concentrations (r = 0.92; p less than 0.01) was observed; a weaker correlation was found between mean arterial pressure and alpha-hANP (r = 0.66) and serum creatinine concentration (r = 0.59), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970178 TI - [Is the plasma ANP level an index of volume expansion in dialysis patients?]. AB - Fluid overload is a frequent complication in anuric patients undergoing hemodialysis (HD) or hemofiltration (HF). Elevated ANP plasma concentrations are associated with overhydration or congestive heart failure (CHF). After intensive ultrafiltration in 18 HD patients with high ANP levels at the end of HD, ANP values normalized (28 +/- 4 fmol/ml) in 11 patients (group A), suggesting previous volume overload, whereas ANP remained elevated (126 +/- 31 fmol/ml) in seven patients (group B). Left ventricular ejection fraction by radionuclide ventriculography (LVEF) was significantly (p less than 0.01) lower in group B (41 +/- 7%) as compared to normal values in group A (67 +/- 8%). M-mode echocardiography demonstrated left atrial enlargement (53 +/- 3 mm) and pathologic enddiastolic left ventricular diameters (58 +/- 4 mm) in group B, compared to normal dimensions of left atrial (43 +/- 1 mm) and left ventricular enddiastolic diameters (47 +/- 4 mm) in group A. Persisting high ANP concentrations after intensive ultrafiltration in HD patients indicate CHF and require further diagnostic evaluation. PMID- 2970179 TI - Atrial natriuretic peptide counteracts angiotensin-II-induced impairment of renal function. AB - The present study was designed to investigate the effect of atrial natriuretic peptide (ANP) on angiotensin II (AT II)-induced impairment of renal function. Female Sprague Dawley were used in this study. After anesthesia and surgical procedure, animals were infused with AT II at two different doses (group I, 15 micrograms/kg BW/min and group II, 15 ng/kg BW/min) into the left renal artery. Subsequently, alpha-ANP was infused intrarenally (left kidney, LK) at 2 micrograms/kg BW/min. Controls received isotonic saline only. In all cases the right and noninfused kidney (RK) served as control organ. In all animals, AT II led to a dose-dependent compromise of kidney function of the LK. Glomerular filtration rate (GFR) had decreased to 0.16 +/- 0.06 ml/min and 0.41 +/- 0.12 ml/min in group I and II, respectively. Subsequent ANP infusion induced a significant increase of GFR to 0.83 +/- 0.23 (p less than 0.01) and 0.99 +/- 0.20 (p less than 0.05) ml/min in group I and II, respectively. Urinary volume (V) exhibited a slight but not significant increase following AT II infusion. ANP infusion led to a highly significant increase of V in both groups. A less pronounced, but significant increase of absolute sodium excretion of the LK was observed in both groups after AT II infusion. Absolute potassium excretion did not change in this phase of the experiment. ANP infusion induced a significant increase in both parameters. Isotonic saline had no effect on all parameters measured. Mean arterial pressure (MAP) had increased dose dependent following AT II infusion. After ANP infusion, a significant diminution of MAP to basal values was obtained from both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970180 TI - [Atrial natriuretic peptide (ANP) in essential hypertension: a humoral marker for salt sensitivity and hypertensive heart disease at a clinically asymptomatic stage?]. AB - Out of 32 patients (f = 10, m = 22; mean age: 46 +/- 3 years) with untreated essential (primary) hypertension (WHO-classification I-III) and without clinical signs of congestive heart failure or chronic renal failure, 19 showed plasma-ANP base levels above the normal range (greater than 100 pg/ml; normal range: 50 +/- 10 pg/ml). While high sodium loading caused an increase of plasma ANP levels and a concomitant decrease of plasma renin and aldosterone concentrations, low sodium loading caused the opposite pattern of ANP and renin/aldosterone secretion. Some patients with essential hypertension with highly elevated plasma ANP levels (10 20-fold above the normal range) showed an only moderate decrease of ANP and a slight increase of renin under a low sodium diet. Plasma ANP levels were significantly correlated with the heart volume (r = 0.54; p less than 0.05; radiologically determined), the electrocardiographic signs of left ventricular hypertrophy (Sokolow-Lyon index; r = 0.62; p less than 0.05) and with the left atrial diameter (r = 0.34; p less than 0.05; determined by 2-D-echocardiography). We speculate that high levels of plasma ANP in patients with essential hypertension might be interpreted as a compensatory mechanism either for an insufficient excretion of sodium or for myocardial dysfunction. PMID- 2970181 TI - [Pathophysiologic and clinical relevance of atrial natriuretic factor in patients with cirrhosis of the liver]. AB - Defects of the ANF system in cirrhosis have not been demonstrated in terms of absolute deficiency of plasma levels or by evidence of major abnormalities of processing in patients with cirrhosis. In the present study, the responsiveness of the ANF system to acute volume stimulation by water immersion and the diuretic and natriuretic effects of ANF infusion were examined. Stimulation of ANF release by 1-h immersion was significantly blunted in ten cirrhotic patients with ascites (increase of plasma ANF by 46 +/- 18%), whereas 11 cirrhotics without ascites showed a 104 +/- 16% increase, similar to the 117 +/- 29% stimulation in 25 healthy controls. Immersion increased urinary volume by 3.6 +/- 0.6/2.0 +/- 0.8/0.7 +/- 0.4 ml/min and urinary sodium excretion by 146 +/- 38/75 +/- 43/43 +/ 19 mumol/min in controls/cirrhotics without/cirrhotics with ascites. Infusion of ANF for 30 min prompted an increase in diuresis and natriuresis in seven cirrhotic patients, which was less marked in patients with ascites, as compared to patients without ascites. Thus, the stimulus response coupling for ANF may be impaired in patients with cirrhosis and ascites. PMID- 2970182 TI - [Skeletal muscle involvement in enthesopathy of the lateral epicondyle of the humerus (so-called tennis elbow)]. AB - Muscle specimens from the origin of the m. extensor carpi radialis brevis, were obtained from 53 patients suffering from chronic enthesopathy of the lateral epicondyle of humerus (EELH). Frank neurogenic atrophy as well as disuse atrophy were caused by radicular irritation due to the cervicobrachial syndrome. The borderline and/or reinnervation patterns of the muscle lesion may be due to the compression neuropathy of the peripheral branches of the radial nerve or may reflect adaptation of the muscle to the chronic nociceptive stimulation in sense of chronic reflex atrophy of the muscle. The elbow joint, and especially the humeroradial joint in cases of EELH represents a functional unit. Any damage to any part leads to secondary reparative and adaptive changes in other structures including the muscle. PMID- 2970183 TI - Asepsis in the dental office. PMID- 2970184 TI - [Facial and skull profile analysis as an aid to orthodontic diagnosis and treatment planning?]. PMID- 2970185 TI - [The Tubingen index for deciduous dentition]. PMID- 2970186 TI - [Incidence and morphological characteristics of frontal crowding in deciduous dentition]. PMID- 2970187 TI - [Overbite bite behavior in the individual from the longitudinal viewpoint]. PMID- 2970188 TI - [Frequency distribution and epidemiology of cheilognathopalatoschisis]. PMID- 2970189 TI - [Reticular atrophy of dental pulp. An artifact or pathomorphological reality?]. PMID- 2970190 TI - [Effect of surface conditioning of sintered ceramic and glass ceramic shells on the compound quality of veneer crowns]. PMID- 2970191 TI - [The value of functional analysis for orthodontic diagnosis]. PMID- 2970192 TI - [The functional requirements of complete dentures]. PMID- 2970193 TI - [The motivation of adults for orthodontic treatment]. PMID- 2970194 TI - [Behavior of T-lymphocytes during the normal menstrual cycle]. AB - FSH, LH, estradiol-17 beta, progesterone, and T-lymphocytes were examined daily in 12 biphasic cycles of 6 healthy women (age 19-29 years). T-lymphocytes were estimated by the alpha-naphthyl-acetate-esterase-reaction as T-cell-marker. There was a significant reduction of T-cells in the first and a nonsignificant reduction in the second cycle phase. The suppressions were related to the high estradiol levels in the follicular as well as the luteal phase. If there is a causative connection and whether the whole immune system is depressed or other regulatory mechanisms interfere isn't quite clear. PMID- 2970195 TI - Further differentiation of Enterobacteriaceae by means of siderophore-pattern analysis. AB - By means of a combination of 5 siderophore bioassays using several indicator strains, genera, species and subspecies of Enterobacteriaceae can be further differentiated. Enterobactin, aerobactin and other siderophores produced can be detected. Each strain shows specific pattern which we called siderophore-pattern. It is easy to separate Morganella, Proteus, Providencia, Yersinia strains from the genera Salmonella, Shigella, Escherichia coli, Enterobacter, Citrobacter, Klebsiella, Serratia. Enterobacter agglomerans strains differ from other Enterobacter species with respect to their siderophore pattern. In Salmonella strains there are differences between the subspecies I and IV. Additionally the most strains of Salmonella subspecies I from nosocomial infections produced aerobactin, in the most cases determined by plasmids. Among Shigella strains different siderophore pattern exist according to other epidemiological markers. S. flexneri strains of serovar 6 produced contrary to the strains of other serovars enterobactin. By means of the siderophore-pattern analysis E. coli strains of serovars 01; 02; 018 can be further differentiated. E. coli 01:K1 strains containing the fimbrial antigen F11 produced aerobactin whereas the F9 strains did not. All Hafnia alvei strains produced a uniform siderophore pattern, different from all other members of the enterobacteriaceae family. With this aim Hafnia alvei strains can be easily separated under practical conditions. PMID- 2970196 TI - [Peridural administration of analgesics using an implanted Rickham reservoir]. AB - Peridurally administered analgetica provide pain relief and may also be used for treatment of chronic pain of malignant origin. Compared to injection via externalized catheter single-shot injection via implantable reservoir and peridural catheter offers some advantages. The authors have used this method first 1985. Technique of implantation and preliminary results of this method are presented as a case report. PMID- 2970197 TI - Complementary use of hysterosalpingography, hysteroscopy and laparoscopy in 100 infertile patients: results and comparison of their diagnostic accuracy. AB - While there is no disputing the invaluable role of laparoscopy in the study of the infertile woman, the function of hysterosalpingography (HSG) and hysteroscopy is still left open to debate. Our research concerns 100 infertile women undergoing HSG, hysteroscopy and laparoscopy; the aim of the study was to check the diagnostic accuracy of the three tests. The sensitivity and specificity of HSG vs hysteroscopy were 74.2% and 91.3% respectively. The sensitivity and specificity of HSG vs laparoscopy were 64.5% and 63.3% as regards tubal patency and 6.9% and 92.8% in the diagnosis of tubal phimosis and pelvic pathology. Despite the limits HSG revealed, our results show that a correct study of the infertile woman can only be accomplished by a combination of all three tests; we therefore recommend a combined approach using all three procedures in all infertile patients. PMID- 2970199 TI - [Percutaneous transluminal angioplasty in peripheral arterial occlusive disease]. AB - Non-surgical recanalization of arterial obstructions has gained world wide acceptance since the development of the balloon dilatation catheter by Gruentzig. The indication for this treatment depends on the clinical stage of the disease, on the location and extension of the occlusion and on the general clinical situation of the patient. Ideal situations with excellent short- and long-term results are stenoses of the iliac arteries and stenoses and short occlusions of the femoropopliteal arteries, whereas treatment of long occlusions of this region is less successful. Definite recommendations concerning additional medical therapy to improve the early and longterm patency rate cannot be given because of lacking studies. The most important advantage of PTA--compared to surgical recanalization--is the lower complication rate. A further improvement of early and late results can be expected by the combined use of different non-surgical recanalization procedures. PMID- 2970200 TI - [Thrombolysis and myocardial infarct]. AB - Thrombolytic therapy in acute myocardial infarction is able to recanalize thrombotic occluded infarct arteries in about 35 to 90%. The results depend on the thrombolytic agent and the route of administration (i.c., i.v.). Successful recanalization causes a reduction of hospital- and one year mortality and reduction of the infarct size in a close correlation to duration of ischemic pain at the beginning of thrombolysis. Significant residual stenoses persist in about 80 to 90% after successful reperfusion. In these patients, PTCA can be performed with high success rates and with a low incidence of complications. Successful PTCA is associated with a reduction of reinfarction and further improvement of left ventricular function. Thus, the value of thrombolysis in acute myocardial infarction in patients with pain duration shorter than 3 to 4 hours is established. The value and optimal point of time for performing PTCA after thrombolysis remains to be defined. PMID- 2970198 TI - Twin studies of coronary heart disease and its risk factors. AB - Traditional twin studies have resulted in higher concordance rates for premature coronary heart disease (CHD) in MZ than in DZ twin pairs. This is in agreement with strong evidence from several other studies, that genetic factors are of importance in the etiology of early onset CHD. Also, in a study of 291 Norwegian twin pairs the concordance rate for hypertension wa 0.36 in MZ and 0.08 in DZ pairs. Relationship between diseases and traditional gene markers have been extensively studied and several associations have been uncovered for CHD. Our group has developed a method to examine a possible permissive or restrictive effect of single genes on the degree of variation that environmental and/or life style factors can cause in a given parameter. This method for studying gene environment interaction is based on the fact that MZ twins are identical with respect to genes, so that any difference between the two members of an MZ pair must necessarily be caused by environmental or life style factors. The possibility that a given gene influences the degree of variability in a parameter such as cholesterol is examined by comparing the within-pair difference in cholesterol level between MZ pairs possessing, and MZ pairs lacking the gene in question, and results of such studies will be presented. New possibilities to study restriction fragment length polymorphisms (RFLPs) at apolipoprotein loci have added a new dimension to research on genetics of CHD and hyperlipidemias. Association between apolipoprotein B, cholesterol and fasting triglyceride levels on one hand and DNA variation at the apolipoprotein B locus on the other has been found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970201 TI - [Percutaneous transluminal coronary angioplasty (PTCA) in the therapy of coronary heart disease]. AB - Percutaneous transluminal angioplasty (PTCA) has been performed worldwide since 10 years and in Graz since 1980. 1977 6 procedures were performed, 1987 about 100,000 will be done worldwide. PTCA today represents a standard treatment of coronary heart disease in addition to pharmacological and surgical therapy. At the present time about one third of patients with coronary heart disease are treated by PTCA. Primary success rate ranges between 80 and 90%, complication rate has dropped under 4%. Besides patients with one-vessel-disease also those with multi-vessel-disease, reduced left ventricular function and patients with unstable angina and acute myocardial infarction are dilated. The "achilles" heel of PTCA is restenosis, which develops in 25 to 35% of cases within 6 months. At present, research is devoted to avoid or prevent this complication. PMID- 2970202 TI - [On behalf of the Austrian Society of Cardiology we are publishing recommendations for using percutaneous transluminal coronary angioplasty (PTCA)]. PMID- 2970203 TI - Paroxetine in the treatment of depression. A double-blind multicenter study versus mianserin. AB - Paroxetine is a new antidepressant drug. It is a potent and selective 5-HT re uptake inhibitor with only weak anticholinergic properties and less effect on the cardiovascular system than the classical tricyclics. In this double-blind multicenter study the antidepressant effect of paroxetine was compared with mianserin in 70 patients with unipolar or bipolar depression. Each drug was administered for 6 weeks after a 1 week run-in period at a daily dosage of 30 mg for paroxetine or 60 mg for mianserin. The 21-item Hamilton Depression Rating Scale (HAM-D) and the physician's global assessment were used to assess efficacy. Both treatment groups showed statistically significant improvement of the HAM-D at Weeks 1 (base-line values: paroxetine mean 28.5; mianserin mean 30.8) through to Week 6 (paroxetine mean 11.5; mianserin mean 17.8) (P less than 0.06). The endpoint differences between treatments however were not statistically different (P = 0.11). The Cleary and Guy factor analysis showed a significant difference (P less than 0.03) at Weeks 2 and 4 for cognitive disturbance and at Weeks 4 and 6 for retardation in favour of paroxetine compared with mianserin. Both drugs were well tolerated with nausea and headache in four patients and somnolence in six patients being reported as the most common side-effect for paroxetine and mianserin respectively. PMID- 2970204 TI - Ca-independent regulation of cardiac myosin. AB - Calcium-independent regulation of the contractile proteins of cardiac muscle has been studied using hyperpermeable cells from rat ventricles and sections of quickly frozen rat hearts. These preparations have been used to study maximum Ca activated force, myosin ATPase activity and the maximum velocity of unloaded shortening. Beta adrenergic activity increases the amount of force and the ATPase activity in accordance with the concentration of the V1 isozyme of myosin. V3 activity is decreased at the same time. In tissues containing only V1, there is no change in maximum velocity in response to beta adrenergic stimulation. These results indicate that beta adrenergic stimulation recruits V1 force generators and probably regulates a transition between a Ca unresponsive and a Ca responsive force generator. A 21,000 dalton protein that reproduces the effect of beta adrenergic stimulation on myosin has been isolated. PMID- 2970205 TI - Cross-bridge kinetics in asynchronous insect flight muscle. AB - Calcium-activated insect flight muscle is further activated by small applied strain. We have investigated the mechanism of strain activation by measuring the extent of oxygen exchange between phosphate and water during ATP hydrolysis by Ca2+ -activated, chemically skinned fibers from the flight muscle of the giant waterbug Lethocerus indicus at different degrees of strain. The maximally activated insect fibres show a pattern of oxygen exchange which is well fitted by a single pathway of ATP hydrolysis. This differs from results with rabbit muscle, which show a more complicated form of oxygen exchange. We ascribe this difference to the different pattern of distribution of myosin heads on the thick filament. Calcium-activated, but unstrained, insect fibres also require more than a simple pathway of ATP hydrolysis to account for the pattern of oxygen exchange. The transition between the more complex pathway and the simple pathway occurs over a very narrow range of strain, centred at zero strain (rest length in the relaxed fibres). We describe how the experimental results of strain activation might occur, based upon Wray's description of filament geometries, but taking into account likely sarcomere mechanics. The pattern of oxygen exchange found in fully activated insect fibres suggests that the process of phosphate release from the actomyosin is rate limiting, implying that the main tension-generating state is an AM.ADP.Pi complex. PMID- 2970206 TI - The elementary steps of the actomyosin ATPase in muscle fibres studied with caged ATP. AB - Caged-ATP (P3-1(2-nitrophenyl) ethyladenosine 5'-triphosphate) has been used to introduce millimolar concentrations of ATP rapidly into glycerinated muscle fibres, thus removing the limit imposed by diffusion to the time resolution of kinetic measurements. We have combined this technique with a rapid freezing method to arrest ATP hydrolysis abruptly. The work of Ferenczi et al. describes the time course of hydrolysis by fibres in the presence and absence of calcium, and for fibres stretched to a length where there is no myofilament overlap. Ferenczi measured the rate of the phosphate burst in muscle fibres. Here we show how these measurements can be extended to measure the equilibrium constant for the hydrolysis step and the rate of ATP release from the active site. Our initial results indicate that in chemically skinned psoas fibres of the rabbit at 12 degrees C, pH 7.1 and ionic strength of 200 mM, the equilibrium constant for the hydrolysis step is 6, and the rate of release of ATP from the active site is 13 s 1. PMID- 2970208 TI - The function of two heads of myosin in muscle contraction. AB - Myosin has two heads which can bind with F-actin and react with ATP. The skeletal muscle myosin forms each 1 mol of the myosin-phosphate-ADP complex (M-P-ADP) and the myosin-ATP complex (M-ATP). The actomyosin ATPase reaction which is coupled with muscle contraction is catalyzed only by the head which forms M-P-ADP. However, the function of M-ATP forming head in muscle contraction has not been elucidated. We studied the binding of S-1 and HMM with F-actin and the dissociation of acto-S-1 or acto-HMM by ATP or AMPPNP using the change in light scattering and fluorescence of pyrene bound to F-actin. S-1 and HMM bound with actin at 1:1 and 1:2 molar ratio, respectively. Acto-S-1 dissociated by one mole of ATP per mole of S-1 but acto-HMM dissociated by 1 mol ATP per mol of HMM (0.5 mol/mol head). Acto-HMM dissociates by AMPPNP (or ADP) via a ternally complex. Acto-HMM bound two mole of AMPPNP, but acto-HMM dissociated by a function of (AMPPNP) but not (AMPPNP)2. These results suggested that the affinity of HMM with F-actin decreased by the binding of one mole of AMPPNP. The result presented here showed that binding of M-ATP forming head with F-actin is controlled by the ATPase reaction of the M-P-ADP forming head. It is suggested that during muscle contraction two heads react cooperatively with thin filament. PMID- 2970207 TI - The role of monovalent phosphate anions in the crossbridge kinetics of chemically skinned rabbit psoas fibers. AB - The role of phosphate ions (Pi) in crossbridge kinetics is investigated by parallel measurements of the ATP hydrolysis rate and the tension transients in maximally activated, chemically skinned rabbit psoas fibers. The tension transients are induced by sinusoidal length oscillations, and resulting time courses are analyzed in terms of three exponential processes (A), (B), and (C). The ATP hydrolysis rate is measured by the NADH fluorscence method coupled with enzymatic reactions which rephosphorylate ADP and oxidize NADH. The hydrolysis rate of the standard activation at 20 degrees C is measured at 0.61 mM/sec. The isometric tension, stiffness, and the ATPase rate progressively decrease with increasing concentrations of Pi (0-16 mM). The decrease is most notably observed with tension, followed by stiffness and the ATPase rate. Both the apparent rate constant and the magnitude parameters of exponential process (B) increase with Pi concentration resulting in a large increase in the oscillatory power output. The active species of Pi which causes this effect is found to be the monovalent anion H2PO4-. The effects of Pi on processes (A) and (C) are only marginal. When fibers are oscillated at 1 Hz, no increase in the ATP hydrolysis rate is observed; a small increase is noticed at 10 Hz (1%), and at 100 Hz (6%). We interpret these results in terms of a crossbridge scheme which adds a branch pathway to the conventional hydrolysis cycle: the number of crossbridges entering the branch pathway increases at higher Pi concentrations and in the presence of oscillations. PMID- 2970209 TI - Cooperative interactions of myosin two heads in muscle force generation. AB - To investigate the possibility of cooperative interactions between the two myosin heads in muscle contraction, Ca2+-activated force development, K+-EDTA- and Mg2+ ATPase activities, muscle fiber stiffness, and the velocity of unloaded shortening were measured on partially p-PDM treated glycerinated muscle fibers, which contained a mixture of myosin molecules with zero, one and two of their heads inactivated. It was found that the magnitude of the Ca2+-activated isometric force development was proportional to the square of both K+-EDTA- and Mg2+-ATPase activities and also to the square of muscle fiber stiffness. If the two myosin heads in the glycerinated fibers are assumed to react independently with p-PDM, the above results strongly suggest that (i) each myosin molecule in the thick filaments can generate force only when its two heads do not react with p-PDM, (ii) muscle fiber stiffness is determined by the total number of native heads, and (iii) there is no cooperative interaction between the two myosin heads in catalyzing ATP hydrolysis. PMID- 2970210 TI - The micromechanics of contraction. AB - This paper suggests the molecular mechanism in muscle whereby some of the delta G of ATP hydrolysis is converted to mechanical work. There is good evidence for thinking that all mechanical, thermal, and chemical observations of muscle result from the summated, repetitive, operation of identical unitary "engines". Such an engine is the S-1 moiety of a myosin crossbridge and the adjacent actin. The operational principle is evident on realizing that S-1 is a particle with two interactive ligand (ATP and actin) binding sites. The change resulting from binding nucleotide is propagated to the actin-binding site, there specifying affinity and angles of attachment. Repetition follows because stepwise enzymatic degradation through intermediates is equivalent to cyclical changing of the site occupant. Trans- (S-1) propagation ensures a work cycle because actin held at a succession of relative positions can generate external work. Proximity mapping and protein chemical studies of S-1 suggest that variation in the position at which actin is held results from the simultaneous operation of a continuing (S-1) actin contact and a polyphosphate charge-modulated coulombic contact. PMID- 2970211 TI - Phosphofructokinase: a component of the thick filament? AB - F-protein, a consistent contaminant of myosin preparations, has been shown to be phosphofructokinase, the key regulatory enzyme of glycolysis. In homogenates of rigor muscle most of the phosphofructokinase sediments with the myofibrils, suggesting that in the living muscle cell phosphofructokinase is not in the soluble fraction as was formerly thought, but bound to the myofibrils. Fluorescent antibody to F-protein labels myofibrils in a zone in each half of the A-band. The increase in separation of the zones across the A-band with increase in sarcomere length suggests that the antibody binds to the parts of the cross bridge regions of the thick filament within the H-zone. It therefore seems likely that phosphofructokinase is located in the cross-bridge region of the thick filament, but that access of antibody is restricted by overlapping thin filaments. PMID- 2970212 TI - Stopped-flow calorimetry of myosin ATP hydrolysis: an implication of chemomechanical energy transduction. AB - Measurements were made of heat production in the pre-steady state phase of ATP hydrolysis by myosin subfragment 1 using an improved stopped-flow calorimeter. This instrument can measure temperature changes as small as 3 X 10(-4) K with a thermal response time of less than 10 ms. When SF-1 was mixed with ATP at 0 degrees C, there was a fairly rapid heat absorbing phase (rate constant, 20-25 s 1 at pH 8) after a burst of heat production was complete within the dead time of the instrument (10 ms). Then followed a phase of very slow heat production with a heat rate of 7 kJ/mol/s. This result indicates that the ATP binding to SF-1 is an exothermic process whereas the subsequent isoenergetic splitting of ATP on the myosin site is endothermic. Thus, a decrease in the internal energy drives the ATP binding which may be regarded as the chemical energy trapping process. By contrast, the myosin intermediate with hydrolysis products (ADP + Pi) is stabilized by a large increase in entropy. In energy transduction by myosin/actin system of muscle, therefore, switching of chemical energy into a form available for mechanical work would be a transition from a low entropy to a high entropy state of weakly actin binding conformations of myosin. PMID- 2970213 TI - [Responsibilities of education in the context of complex rehabilitation]. PMID- 2970214 TI - [Retinal circulatory changes in diabetic retinopathy]. PMID- 2970215 TI - The Doppler signal: where does it come from and what does it mean? AB - Doppler sonographic measurement of blood velocity and associated physiologic parameters is a powerful diagnostic technique. State-of-the-art instrumentation incorporates velocity measurement with two-dimensional imaging capability; it uses intensity and color coding to display complex physiologic and anatomic data to the observer in an easily understood format. Although the concepts underlying Doppler sonography are not complex, mastery of the technique requires extra training and commitment. The principal features and clinical practicalities associated with Doppler sonography are summarized in the following paragraphs. Continuous-wave Doppler is very sensitive to small vessels and has no upper velocity limit. In addition, the instrumentation is not complex and produces relatively low acoustic power. A significant drawback to continuous-wave Doppler is that there is no depth sensitivity, and thus complex structures or multiple vessels can give conflicting information. Pulsed Doppler (including duplex and color-flow) instrumentation has the capability of depth resolution and a variable sample volume. Pulsed Doppler equipment is prone to aliasing (false velocity indications) under some circumstances and also produces higher peak power levels than does continuous-wave equipment. Duplex equipment is more complex and expensive than continuous-wave equipment because the two-dimensional and Doppler modes must be synchronized in operation and display. Color-flow equipment is extremely complex and expensive. Color flow provides information of a qualitative and limited quantitative value. Absolute measurement still requires range-gate measurements. Technical and anatomic factors will affect the measured velocity profiles. Thus, it is important to fully appreciate the anatomy of the vessel and the angle between the vessel and the ultrasound beam when making quantitative measurements. Measurements that evaluate the velocity waveform and make use of ratios, such as the pulsatility index, eliminate the need for angular corrections; however, artifacts due to unappreciated anatomic or wall characteristics may lead to incorrect information if all parameters are not fully understood. Doppler sonographic measurements may be used to determine the presence of flow, determine the direction of flow, identify time-varying velocity characteristics, and detect velocity disturbances. Because flow and velocity are related, it is possible to estimate flow from velocity measurements with careful calibration and proper precautions. Velocity is related to flow, which, in turn, is related to both pressure and vascular resistance.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970216 TI - Focal renal artery stenosis caused by fibromuscular dysplasia: treatment by percutaneous transluminal angioplasty. AB - We performed percutaneous transluminal angioplasty of the renal artery in five hypertensive patients with focal renal artery stenoses caused by fibromuscular dysplasia. In four patients, the hypertension decreased or resolved. In all patients, the stenoses displayed considerable resistance to dilatation, requiring maximum inflation of the angioplasty balloon with 10 atm of pressure (10.1 x 10(5) Pa). In two patients, the stenoses were extremely firm with a persistent waist noted in the maximally inflated balloon. One of these patients was left with a residual 40-50% stenosis after initial angioplasty, and a second attempt at dilatation after restenosis was also unsuccessful. This patient's blood pressure did not improve. When the results of our experience in these five patients were combined with 22 cases reported in the literature, we found that 23 (85%) of the 27 patients with focal renal artery stenoses caused by fibromuscular dysplasia had a decrease in blood pressure after percutaneous dilatation. Percutaneous transluminal angioplasty is an effective treatment for patients with renovascular hypertension caused by focal renal artery stenoses resulting from fibromuscular dysplasia. PMID- 2970217 TI - Management of acne vulgaris. PMID- 2970218 TI - High-dose medroxyprogesterone acetate plus mitomycin-C or vindesine in the treatment of advanced breast cancer. AB - Forty women with metastatic breast cancer were randomized to receive either mitomycin-C (MMC) (21 patients) or vindesine (19 patients) with high dose medroxyprogesterone acetate (MPA). Response rates were not significantly different (33% for MPA and MMC and 28% for MPA and vindesine); the duration of response was better for patients receiving the MMC combination (median of 10.5 months versus 6 months for those on the vindesine combination). PMID- 2970219 TI - Intentions to use predictive testing by those at risk for Huntington's disease: implications for prevention. AB - Huntington's disease (HD) is a late onset, genetic disorder that is incurable and undetectable until the onset of symptoms. A marker for the gene that causes HD was recently discovered which will lead to a predictive test. The purpose of this research was to assess the knowledge, attitudes, beliefs, and behavioral intentions concerning the impending predictive test by those at risk for HD. Results indicated that a majority of participants favored using the test presymptomatically and would encourage their adult children to use it as well. Fewer participants favored using the predictive test prenatally or on minors. Characteristics that best differentiated those who intend and do not intend to use predictive testing were identified and discussed. Suggestions for interventions were presented with an emphasis on outreach and prevention programs designed to prepare HD families for the difficult decisions the predictive test will precipitate and the subsequent impact of those decisions. PMID- 2970220 TI - Guiding individual decisions: a randomized, controlled trial of decision analysis. AB - In early 1983, all 1,280 faculty and resident physicians at one hospital who were eligible to be vaccinated against hepatitis B were divided randomly into three groups: Group 1 physicians received general information about the risks and benefits of alternative vaccine decisions; Group 2 physicians were additionally invited to provide personal information for an individualized decision analysis (12.6 percent responded); and Group 3 physicians, who served as controls, were not contacted. In one year's follow-up, 20 percent of physicians were screened for hepatitis B antibody or vaccinated. More Group 2 physicians whose decision analyses recommended screening or vaccination took these actions (39 percent) than any other group. Group assignment remained significantly associated with vaccine decisions after analyzing results by the "intention to treat" principle, and after adjusting for training status, exposure to blood and blood products, and pre-study intentions about the vaccine. Despite the low overall vaccine acceptance rate, it is concluded that individualized decision analysis can influence the clinical decisions taken by knowledgeable and interested patients. PMID- 2970221 TI - Pregnancy after traumatic aortic aneurysm repair. AB - Thoracic aortic aneurysm is associated with increased morbidity and mortality in pregnancy. Successful pregnancy with vaginal delivery after repair of a traumatic aortic aneurysm is described. PMID- 2970222 TI - The influence of body weight on lipoprotein lipids in patients with polycystic ovary syndrome. AB - Lipoprotein lipid and androgen profiles were compared after a 12-hour fast that involved 13 women with polycystic ovary syndrome and 13 who did not have the syndrome. All women were matched for percent ideal body weight. As expected, patients with polycystic ovary syndrome had significantly higher luteinizing hormone--to--follicle-stimulating hormone ratios, higher testosterone levels, higher free testosterone levels, higher dehydroepiandrosterone sulfate levels, and lower testosterone-estradiol-binding globulin binding capacity than the other women. Patients with polycystic ovary syndrome had significantly higher mean serum triglyceride levels, very-low-density lipoprotein cholesterol levels, and lower high-density lipoprotein cholesterol levels. Differences in body weight do not explain the male pattern of lipoprotein lipid concentrations in women with polycystic ovary syndrome. The association of hyperandrogenism with lipoprotein lipid abnormalities should be evaluated in regard to the influence of endogenous sex steroids on differences between the sexes in incidence of cardiovascular disease. PMID- 2970223 TI - C19 adrenal steroids enhance prostaglandin F2 alpha output by human endometrium in vitro. AB - Dehydroepiandrosterone sulfate significantly increased prostaglandin F2 alpha output by fragments of human secretory endometrium during the first and second 24 hour periods of incubation in Ham's F-10 medium containing 10% charcoal-treated calf bovine serum. The effects of dehydroepiandrosterone sulfate were noted at 10(-6) mol/L concentrations, which are close to the normal plasma levels of this compound. For the purpose of comparison, the effects of estradiol at 10(-8) mol/L and those of unconjugated dehydroepiandrosterone, dehydroepiandrosterone acetate, 5-androstene-3 beta, 17 beta-diol, or 5 alpha-dihydrotestosterone were evaluated at 10(-8) to 10(-8) mol/L concentrations in parallel experiments. All of the delta 5-C19 steroids tested enhanced prostaglandin F2 alpha output at 10(-6) mol/L but not at 10(-8) mol/L; 5 alpha-dihydrotestosterone was inactive at 10(-6) mol/L but showed a stimulatory effect at 10(-5) mol/L in two experiments. The stimulation of prostaglandin F2 alpha production by the adrenal steroids was significantly reduced by the antiestrogen 4-hydroxytamoxifen at 10(-6) mol/L (a finding consistent with the reported utilization of the estrogen receptor for their actions on other systems) but was not affected by the antiandrogen 4 hydroxyflutamide at 10(-6) mol/L. Progesterone (10(-7) mol/L) also lowered the effects of dehydroepiandrosterone (10(-6) mol/L) and 5-androstene-3 beta,17 beta diol (10(-6) mol/L), as well as those of estradiol (10(-8) mol/L). delta 5-C19 steroids at 10(-6) mol/L levels did not antagonize the effect of 10(-8) mol/L estradiol, whereas 5 alpha-dihydrotestosterone reduced it by 50% at these concentrations. The significant effect of 5 alpha-dihydrotestosterone points to potential antiestrogenic effects of the C19 compounds that may be manifested in vivo at particular C19/estradiol concentration ratios. The demonstration of direct estrogenic effects of delta 5-C19 steroids, which are not significantly converted to estrogens in vivo, justifies their use in estrogen replacement preparations and indicates that aromatase inhibitors may not eliminate completely the stimulation of estrogen-responsive breast and endometrial tumors by the patient's adrenal steroids. PMID- 2970224 TI - A critical comparison of results of endolymphatic subarachnoid shunt and endolymphatic sac incision operations. AB - Detailed information was obtained by questionnaires, office visits, chart reviews, and telephone calls from 171 of 179 consecutive patients who underwent endolymphatic sac surgery between 1976 and 1986. A comparison of surgical results was made. For patients followed more than 24 months, complete control of vertigo was obtained in 40 of 68 patients (59%) by endolymphatic subarachnoid shunt and in 27 of 60 patients (45%) who underwent a variety of sac incision operations. This difference achieved borderline statistical significance (p less than or equal to 0.057). Profound or total hearing loss occurred in 8 patients (4.3%), 6 of whom had sac mastoid procedures. The percentage of patients indicating an absence of disability improved in both groups dramatically following surgery. PMID- 2970225 TI - Sodium balance effects on renin, angiotensinogen, and atrial natriuretic polypeptide mRNA levels. AB - The expressions of the renin, angiotensinogen, and atrial natriuretic polypeptide (ANP) genes were studied in rats with varying sodium intakes. mRNAs for angiotensinogen and renin were measured by a sensitive radiodensitometric hybridization assay. ANP mRNA was determined by a dot-blot hybridization assay. In the high-sodium state, plasma renin concentration (PRC), renal renin concentration (RRC), and renal renin mRNA decreased by 88, 90, and 75%, respectively. In the low-sodium state, PRC, RRC, and renin mRNA increased 17 fold, 2.5-fold, and 4.5-fold, respectively. With captopril treatment during sodium depletion, PRC and renin mRNA increased 144-fold and 17.1-fold, respectively, and RRC increased 4.2-fold. The angiotensinogen mRNA level in the liver and kidney decreased in the high-sodium and increased in the low-sodium state to similar extents. Captopril treatment decreased angiotensinogen mRNA in both organs by about half. Plasma ANP and ANP mRNA decreased in the low- compared with the high-sodium state, whereas atrial ANP content was unaltered in both low- and high-sodium states. These results demonstrate that sodium intake affects the expression of the renin and angiotensinogen genes and slightly alters the expression of ANP gene. PMID- 2970227 TI - Pancreatic islet discrimination of hexose anomers. I. Steady-state computer simulation. AB - Pancreatic islets detect glucose level by phosphorylating it and converting the glycolytic rate to a signal to secrete insulin. Insulin secretion is greater from the alpha- than from the beta-anomer when the D-glucose level is below 22 mM. D mannose behaves similarly but at nearly twofold higher concentrations. Two explanations have been proposed: 1) glucokinase, which has the same anomeric preference, is the principal hexose phosphorylating enzyme and limits glycolytic rate. 2) Phosphofructokinase limits glycolysis and hexokinase is the principal enzyme phosphorylating hexose; hexosediphosphate activators of phosphofructokinase are more readily synthesized from alpha-anomers of hexose phosphates. We have simulated both alternatives with a detailed anomerically specific model of the hexose-metabolizing glycolytic enzymes. The pathway preference for alpha-anomer of both hexoses was adequately reproduced with anomerically active limiting glucokinase. The other mechanism did not reproduce the observed pathway preference. PMID- 2970226 TI - Phenylarsine oxide and denervation effects on hormone-stimulated glucose transport. AB - Insulin and insulin-like growth factor I (IGF-I) stimulate glucose transport in skeletal muscle through separate receptors. The proximal postreceptor events in coupling insulin and IGF-I receptors to glucose transport have been suggested to differ. Denervation of skeletal muscle produces a postreceptor insulin resistance presumably at an early step in the signaling cascade. We examined the effects of denervation and phenylarsine oxide (PAO), an agent believed to block insulin action on transport at a postreceptor step, on insulin and IGF-I stimulated 2 deoxy-D-glucose transport in isolated solei. Denervation (24 h) produced severe IGF-I resistance without affecting IGF-I receptor number or affinity. PAO inhibited insulin and IGF-I stimulation of transport in control muscles by approximately 90 and approximately 70%, respectively. In denervated muscle PAO inhibited transport stimulation by both hormones less than in controls. Conclusions are that 1) skeletal muscle insulin and IGF-I receptors signal transport mainly through a PAO-sensitive mechanism, but IGF-I's action involves a larger PAO-resistant component; 2) the denervation-induced postreceptor resistance of glucose transport to both hormones involves primarily the PAO sensitive pathway. PMID- 2970228 TI - Continuous monitoring of portal vein and hepatic artery hemodynamics in unrestrained rats. AB - Continuous measurement of portal vein and hepatic artery blood flows during physiological experimental conditions, even in large animals, poses difficult problems. We report the successful use of miniaturized flow probes and pulsed Doppler flowmetry for chronic monitoring of hepatic artery and portal vein flows in intact unrestrained rats and describe the probe construction and implantation. Proportionality between portal vein velocity and portal flow was made possible by a technique for stabilizing the diameter of the venous segment from which velocity is recorded. The accuracy of the method in detecting changes in portal vein flow was established by the high statistically significant correlation between changes in velocity recorded simultaneously from portal vein and superior mesenteric artery in a series of rats with ligated celiac and inferior mesenteric arteries. In these preparations all portal vein flow is derived from the superior mesenteric artery. Complex dynamic changes in the hepatic circulation of conscious unrestrained rats were recorded in response to systemic injections of glucagon and angiotensin II. In the resting state several characteristic velocity patterns were recorded from the portal vein. Oscillations linked to respiration were not observed while the animals rested quietly but were noted during sleep and anesthesia. Two hitherto unrecognized patterns produced respectively by the pulsations of the superior mesenteric artery and by spontaneous contractions of the portal vein were also observed. The method described here provides the first opportunity to study hepatic circulation in chronically instrumented rats during physiological experimental conditions. PMID- 2970229 TI - Tubular site(s) of action of atrial natriuretic peptide in the rat. AB - With micropuncture techniques, the present study examined the delivery of chloride to the superficial late distal tubule and the base and tip of the papillary collecting duct in rats treated with either Wy 47663, a synthetic analogue of atrial natriuretic peptide (ANP), or vehicle alone. Whole kidney glomerular filtration rate (GFR) and single-nephron glomerular filtration rate were not significantly different between the two groups. Late distal tubule chloride delivery was also not different between ANP- (5.71 +/- 1.15%) and vehicle- (6.28 +/- 1.12%) treated animals. However, fractional delivery to the base of the papillary collecting duct was significantly greater in the ANP treated rats (14.37 +/- 1.98%) compared with vehicle-treated rats (7.32 +/- 1.47%). Tip papillary collecting duct delivery was also significantly greater in the ANP-treated rats (1.97 +/- 1.96 vs. 3.09 +/- 0.60%). In addition, the percent of chloride delivered that was reabsorbed along the papillary collecting duct was significantly less in the ANP-treated rats. In conclusion, ANP inhibits reabsorption in some tubular segments between the superficial late distal tubule and papillary collecting duct base as well as in the accessible portion of the papillary collecting duct. PMID- 2970230 TI - Effect of sodium intake on gene expression and plasma levels of ANF in rats. AB - The effect of short- and long-term sodium loading and sodium restriction on the gene expression as well as on circulating plasma levels of atrial natriuretic factor (ANF) was evaluated in normotensive Wistar rats. These rats were fed either a low-, a regular-, or a high-sodium diet (regular diet and 1% saline as drinking fluid) and studied after 1 and 3 wk. The ANF mRNA was determined in pooled atria and ventricles of the different groups of rats, using the dot-blot technique. Plasma ANF levels were measured with a radioimmunoassay. After 1 wk on the high-sodium diet, ANF mRNA was increased in right atria and ventricles together with circulating ANF levels when compared with animals maintained for the same period on a low-sodium diet. After 3 wk on the various diets, the differences in cardiac ANF mRNA and in plasma ANF levels had disappeared. Gene expression of ANF was also looked for in different areas of the brain, lung, thyroid, adrenals, and the kidney; no hybridization was detected in any of these organs. These data suggest that in rats, the transcription of the ANF gene and peptide release in enhanced only during short-term adaptation to dietary sodium loading. PMID- 2970231 TI - Direct cardiac effects of vasopressin: role of V1- and V2-vasopressinergic receptors. AB - Experiments were performed to determine the possible direct effects of arginine vasopressin (AVP) on cardiac function in the nonworking Langendorff preparation. Hearts were isolated from male Wistar rats, and the coronary arteries were retrograde perfused at a constant rate through the aorta with a Krebs-Henseleit solution, which was continuously bubbled with 95% O2-5% CO2. The hearts were paced at 280 beats/min and measurements made of peak ventricular pressure (PVP), first derivative of left ventricular pressure (dP/dtmax), and coronary perfusion pressure (CPP). By maintaining constant coronary flow, the direct cardiac effects of AVP could be determined independent of changes in myocardial O2 delivery elicited by potential coronary vasoconstriction. Myocardial function was assessed at AVP concentrations of 0, 10, 25, 50, 100, 200, 400, and 500 pg/ml. Progressive coronary vasoconstriction was observed with increasing AVP concentration. In contrast, PVP and dP/dtmax increased at 50 and 100 pg/ml of AVP but fell at 400 and 500 pg/ml. The maximal PVP and dP/dtmax responses were at 50 pg/ml (+16 +/- 3 and +44 +/- 4%, respectively), whereas at 500 pg/ml both PVP and dP/dtmax were reduced below control (-30 +/- 4 and -34 +/- 5%, respectively). Pretreatment with the specific V1-vasopressinergic antagonist d(CH2)5Tyr(Me)AVP (40 ng/ml) totally blocked both the coronary vasoconstrictor and contractility responses to AVP. Furthermore, infusion of a specific V2-agonist was without effect even at high doses. These data suggest that although AVP causes dose-related coronary vasoconstriction over a wide range of AVP concentrations, the hormone may exert a positive inotropic effect at doses mimicking circulating levels encountered in a number of pathophysiological situations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970232 TI - Acute hypoxemia stimulates atrial natriuretic factor secretion in vivo. AB - The hypothesis was tested that acute hypoxemia may be a physiological stimulus for atrial natriuretic factor (ANF) secretion in anesthetized, spontaneously breathing rabbits. Base-line plasma ANF (range from 29.8 to 219 pg/ml; mean +/- SE = 87.0 +/- 14.1 pg/ml; n = 16 rabbits) was negatively correlated with base line arterial PO2 (r = -0.759; P less than 0.01) but not with PCO2, pH, mean arterial blood pressure, central venous pressure (CVP), minute ventilation, heart rate, or type of anesthetics used. Acute hypoxemia (arterial PO2 22.3-44.3 mmHg) lasting 10 min increased plasma ANF levels over base line by 69.2 +/- 47.7 (SE) pg/ml at 6 min and 87.5 +/- 46.8 (SE) pg/ml at 9 min (P less than 0.01; n = 9). The increase in arterial pH and minute ventilation and the decrease of arterial PCO2 paralleled the changes in plasma ANF. Mean arterial blood pressure, CVP, and heart rate did not change significantly. ANF responses to hypoxemia (range from 4.4 to 423 pg/ml) correlated with base-line CVP (r = 0.761; P less than 0.01) and base-line ANF (r = 0.523; P less than 0.05) but with no other measured variable. Although the mediators of hypoxemia-induced release of ANF need to be explored further, this study raises the possibility that ANF might be involved in the adaptation to hypoxemia. PMID- 2970233 TI - Cardioprotective actions of thromboxane receptor antagonism in ischemic atherosclerotic rabbits. AB - Atherosclerosis was induced in New Zealand White rabbits by feeding them a 0.5% cholesterol-enriched rabbit chow for 10-12 wk. Half of the cholesterol-fed rabbits were given BM 13505, a specific thromboxane A2/endoperoxide (TxA2/PGH2) receptor antagonist, and the other half were given its vehicle (i.e., 2% Na2CO3). At the end of 10-12 wk, the rabbits underwent experimental myocardial ischemia or an identical sham operation, except that the coronary artery was not occluded. BM 13505 was shown to protect the ischemic rabbit myocardium by three different methods: 1) maintenance of myocardial tissue creatine kinase (CK) activity in the ischemic myocardium; 2) reduced loss of free amino nitrogen-containing compounds from the myocardium; and 3) blunting the rise of plasma CK activity. Part of the mechanism for these effects may be due to inhibition of platelet aggregation and blockade of the vasoconstrictor effect of TxA2. However, these protective effects were not due to differences in myocardial oxygen demand among the groups. Finally, BM 13505 exhibited an antiatherogenic effect by reducing the deposition of cholesterol in the aortic wall and by retarding plaque formation in coronary arteries. However, it does not achieve this antiatherogenic effect by lowering plasma cholesterol concentrations or by scavenging superoxide free radicals. Thus blockade of TxA2 receptors exerts a variety of beneficial effects that reduce the severity of ischemic damage resulting from myocardial ischemia. PMID- 2970234 TI - Atrial distension of isolated rabbit hearts and release of atrial natriuretic factor. AB - Interventions that increase atrial pressures in humans or laboratory animals release atrial natriuretic factor (ANF) into the circulation. We studied the relation between distension of the right or left atrium and release of ANF in retrograde-perfused isolated rabbit hearts. A fluid-filled balloon within the right or left atrium was inflated to a mean pressure of 5, 10, 15, or 20 mmHg, and ANF in the cardiac effluent was measured by radioimmunoassay. The slope of the regression line relating ANF release to atrial distending pressure was steeper for the left than right atrium (P less than 0.001), indicating that, at comparable increases in mean pressures, the left atrium releases more ANF than does the right atrium. Left atrial tissue concentration of ANF was greater than right atrial (1.58 +/- 0.15 vs. 1.05 +/- 0.09 micrograms ANF/mg protein, P less than 0.01). In contrast to previous studies showing right atrial dominance in rats, the left atria of isolated, perfused rabbit hearts contain more ANF and release more in response to atrial distension. PMID- 2970235 TI - Natriuresis during atrial distension and a concurrent decline in plasma atriopeptin. AB - In the conscious dog, left atrial distension elicits a composite response that modulates both cardiovascular and renal function. The response to atrial distension may be mediated by the combined effects of neural reflexes and the release of atriopeptin. To assess the relative contributions of atrial reflex mechanisms and circulating atriopeptin to the renal response elicited by atrial distension, alpha-human atrial natriuretic peptide (alpha-hANP) was infused into conscious dogs at 50 ng.kg-1.min-1 for 60 min. Then the infusion was stopped abruptly, and left atrial pressure was increased 8 mmHg by inflating a balloon positioned above the mitral valve. Plasma atriopeptin decreased during the 40-min period of atrial distension, but urine flow and sodium excretion increased during this time. In another series of experiments, volume expansion was substituted for atrial distension. Saline (24 ml/kg) was infused intravenously for 5 min immediately after the 60-min period of alpha-hANP infusion. Urine flow and sodium excretion increased after administration of saline even though plasma atriopeptin decreased substantially during the same time period. These results provide evidence that circulating levels of atriopeptin do not play a dominant role in influencing sodium excretion either during atrial distension or in response to saline infusion. PMID- 2970236 TI - Anaphylactic reaction to isoflurane. PMID- 2970237 TI - Histochemical types and sizes of fibers in the rectus abdominis muscle of guinea pig: adaptive response to pregnancy. AB - Effects of pregnancy stimulation upon histochemically assessed myofibrillar ATPase and muscle fiber diameters were analysed in the rectus abdominis (RA) muscle of guinea pig. Samples of the muscle were taken at 30, 40, 50, 60, and 70 days of pregnancy and compared with samples of the same muscle taken from nonpregnant guinea pigs. Changes in muscle fiber proportions were noted through the course of pregnancy. Starting from 50 days of gestation an increase in type I fibers and a decrease in type IIB fibers were noted. Increase in muscle fiber diameters was also observed in type I, IIA, and IIB fibers. In addition, the RA muscle of the male guinea pig was compared with that of the female guinea pig and showed more type IIA and less type IIB fibers and all the three fiber types were larger than those of the female. PMID- 2970238 TI - The influence of human immunodeficiency virus (HIV) infection on antibody responses to influenza vaccines. AB - STUDY OBJECTIVE: To ascertain whether subjects infected with human immunodeficiency virus (HIV) generally develop protective hemagglutination inhibition antibody responses to inactivated influenza vaccines. DESIGN: Prospective study of 104 persons before and after immunization. SETTING: Outpatient clinic and hospital ward. PATIENTS: Persons with the acquired immunodeficiency syndrome (AIDS) (n = 25), AIDS-related complex (n = 14), and HIV seropositive men with only lymphadenopathy or no symptoms (n = 27). Controls were HIV-seronegative homosexual men (n = 22) and HIV-seronegative heterosexuals (n = 16). INTERVENTIONS: Subjects were immunized with inactivated vaccines containing 15 micrograms of each of the following influenza virus hemagglutinins: A/Taiwan/1/86 (HINI), A/Mississippi/1/85 (H3N2), A/Chile/1/83 (HINI), and B/Ann Arbor/1/86. MEASUREMENTS AND MAIN RESULTS: Fourfold or greater antibody responses occurred less frequently in subjects with HIV infections than in HIV-seronegative controls. Protective levels (1:64 or greater) of hemagglutination inhibition antibodies were attained by 94% to 100% of HIV-seronegative controls, 52% to 89% of HIV-seropositive asymptomatic subjects, and 13% to 50% of subjects with AIDS or AIDS-related complex. No increase in the prevalence or level of serum HIV p24 antigen or clinical deterioration was detected among HIV-infected persons after influenza immunization. CONCLUSIONS: Because of the poor antibody responses to influenza vaccines among HIV-infected subjects, even in many with no or minimal symptoms, alternative strategies for preventing influenza, such as booster doses of influenza vaccine, prophylaxis with amantidine, or both should be considered. PMID- 2970239 TI - [Severe psoriasis induced by chloroquine (nivaquine)]. AB - The adverse effects of a variety of drugs on psoriasis are well documented, and they generally result in aggravation of the disease. We report the case of a 75 year old man who developed a psoriatic erythroderma and a generalized pustular psoriasis two weeks after the beginning of an antimalarial treatment. The psoriasis responded poorly to etretinate alone, and an etretinate-methotrexate combination was needed to cure the patient. Some psoriasis vulgaris plaques persisted for six months after the acute complication. Thus, chloroquine should be entered in the list of drugs which may induce psoriasis. PMID- 2970240 TI - ["Crocodile baby", a non-fatal form of malignant keratoma?]. PMID- 2970241 TI - Treatment of abdominal nerve entrapment syndrome using a nerve stimulator. AB - Seventy-six patients treated at York Pain Relief Clinic for Abdominal Nerve Entrapment Syndrome (ANES) between 1982 and 1986, using aqueous phenol and nerve stimulator control are reviewed. A questionnaire was sent to all the patients who had been discharged from the clinic to try to confirm that the initial improvements had been maintained and 60 patients replied. Group A (n = 44) had been diagnosed with confidence; 95% had gained complete or partial relief of symptoms. Group B (n = 32) had other symptoms making the diagnosis less certain; 50% gained some relief. Clinical presentation of ANES and the method of treatment are described. PMID- 2970242 TI - Diagnosis of significant abdominal trauma after road traffic accidents: preliminary results of a multicentre clinical trial comparing minilaparoscopy with peritoneal lavage. AB - A prospective multicentre study comparing the value of the recently introduced minilaparoscopy with peritoneal lavage in patients with abdominal trauma is in progress. To date 55 patients with blunt abdominal trauma have been entered into the study. Following initial resuscitation, 26 were randomised to peritoneal lavage and 29 to minilaparoscopy performed under intravenous sedation and local anaesthesia. The two groups were comparable with respect to age, sex, incidence of multiple injuries and mortality (2 patients in the lavage group and 1 in the minilaparoscopy group). A negative test was obtained in 15 patients subjected to lavage and 12 patients who underwent minilaparoscopy. A further four patients in the minilaparoscopy group were found to have a minimal static haemoperitoneum. All these patients were treated conservatively and none required surgical intervention on the abdomen. Thus neither investigation carried a false negative rate. A positive test was obtained in 11 patients in the lavage group and significant findings were observed in 13 patients assessed by minilaparoscopy. All these patients were subjected to emergency laparotomy. Absence of significant bleeding or trauma was observed at laparotomy in 3/11 (27%) and 1/13 (8%) in the lavage and minilaparoscopy groups respectively. Although both procedures were highly sensitive for the detection of significant intra-abdominal injury (100%), the specificity was 83% for peritoneal lavage and 94% for minilaparoscopy. The predictive value of a positive minilaparoscopic examination was 92% as opposed to a positive predictive value of 72% for peritoneal lavage. PMID- 2970243 TI - Contribution of education to cost effective care of health maintenance needs, low back pain, abdominal pain, and systemic hypertension. PMID- 2970244 TI - [Coronary angioplasty in the elderly patient. Immediate results and mid-term outcome]. AB - Between April 1980 and July 1986, 50 patients over 65 (32 men and 18 women; mean age: 72 years) were treated by transluminal coronary angioplasty. Before the dilatation, 58 p. cent of the patients presented a severe angor (class III or IV) and 44 p. cent were multi-truncular. The dilatation was successful in 39 patients (78%) and 13 patients developed a recurrent stenosis successfully treated in 9 instances by re-dilatation. Among the complications, there were 2 deaths (4%), 3 infarctions (6%) and 3 emergency coronary bypass operations (6%). With a mean 28 months follow-up, the overall survival is 92 p. cent. After a successful dilatation, 49 p. cent of the patients are completely asymptomatic and the subsequent cardiological hospitalizations are rare (11%). These results show that coronary dilatation represents an interesting therapeutic option in elderly coronary patients. PMID- 2970245 TI - [Revascularization in the acute phase of myocardial infarction. Study of left ventricular function in the 3d week and 6th month]. AB - 27 patients with primary myocardial infarction are treated, in the first 6 hours, with intravenous thrombolysis with immediate coronary angiographic control showing a patent artery in 76 p. cent of the cases. The treatment is completed with intracoronary thrombolysis (5 times) and transluminal angioplasty (6 times), enabling to obtain a patency rate of 88 p. cent. The overall left ventricular function is evaluated at D21 and in the 6th month (M6). Among anterior infarctions, those treated during the first three hours have a better ejection fraction (EF) at D21 than those treated later (40.3 p. cent +/- 6 versus 33.2 p. cent +/- -NS); this functional benefit is confirmed at M6 (42.8 p. cent +/- 12 versus 30.6 p. cent +/- 8; p less than 0.06). On the contrary, among inferior infarctions, the EF is comparable at D21 and M6 regardless of the early nature of the treatment. One patient died prematurely from a cerebral vascular accident which occurred during the thrombolysis. These results are in favor of a significant myocardial salvation in anterior infarctions revascularised at an early stage and of an active approach in order to obtain the most complete possible revascularisation. PMID- 2970246 TI - Immunology of renal transplantation in Singapore. AB - Renal transplantation is the best form of renal replacement therapy for patients suffering from endstage renal failure. While correcting the metabolic consequences of uremia, it is the only form of replacement therapy which permits full rehabilitation of the patient. However, the successful engraftment of a renal allograft is frequently limited by immunologically mediated graft loss which can be as high as 50% in the first year in cadaveric transplants. Since the beginning of the history of transplantation, various methods of modifying allograft rejection, both pretransplantation and posttransplantation have been studied. In this paper, we report the pretransplantation immunological factors that appear to modulate graft survival in renal transplantation in Singapore. PMID- 2970248 TI - Differential cognitive impairment in Alzheimer's disease and Huntington's disease. AB - The differentiation between cortical and subcortical dementias requires that the cognitive characteristics of dementias attributable to different causes be discriminable. For large samples of Alzheimer's disease and Huntington's disease patients, distinct cognitive profiles were obtained on the Mini-Mental State Exam. The profile differences were independent of severity of dementia and were sufficiently robust to classify patients as Alzheimer's disease or Huntington's disease with 84% accuracy. The qualitative differences in cognitive functioning may also be typical of other cortical and subcortical dementias. PMID- 2970247 TI - Positron emission tomographic scan investigations of Huntington's disease: cerebral metabolic correlates of cognitive function. AB - Fifteen drug-free patients with early to mid-stage Huntington's disease (HD) were evaluated with positron emission tomographic (PET) scans of 18F-2-fluoro-2-deoxy D-glucose uptake and quantitative measures of neurological function, learning, memory, and general intelligence. In comparison with a group of normal volunteers, the HD patients showed lower metabolism in both caudate (p less than 0.001) and putamen (p less than 0.001) on PET scans. A significant and positive relationship was found between neuropsychological measures of verbal learning and memory and caudate metabolism in the patient group but not in the normal group. Visual-spatial learning did not reflect a similar pattern, but performance intelligence quotient was positively related to both caudate and putamen metabolism in the HD group. Vocabulary level was unrelated to either brain structure. Discussion focuses on these and other observed brain-behavior relationships and on the implications of these findings for general behaviors such as those involved in coping and adaptation. PMID- 2970249 TI - [Congenital heart diseases and obstructive pulmonary vascular diseases in Down's syndrome. Apropos of 142 children with trisomy 21]. AB - Trisomy 21 accounts for 3 p. 100 of reasons for admission to the Paediatric Cardiology unit of the St Luc University Clinics, Brussels. In a series of 142 cardiac children with trisomy 21 evaluated by catheterization between 1969 and 1987, 54 p. 100 of the cardiac malformations observed consisted of persistent common atrioventricular canal (complete in 45 p. 100 of the cases). The other heart diseases were ventricular septal defect (23 p. 100), atrial septal defect of the ostium secundum type (10 p. 100) and tetralogy of Fallot (9 p. 100). In 40 p. 100 of the patients other cardiovascular abnormalities were associated with these predominant intracardiac shunts. These findings were in agreement with those usually reported in the literature. At the time of investigation (mean age 24 months), pulmonary vascular resistance had already reached a pathological level in 88 p. 100 of the cases. Oxygen tests only slightly improved these results, which suggested that the conditions were favourable to the early development of a pulmonary obstructive vascular disease in Down's syndrome, thus darkening the prognosis of congenital heart disease in mongoloid children. In this series to overall mortality rate of corrective surgery was 23 p. 100. The risk was maximum in infants aged less than 3 months with severe and rapidly symptomatic lesions. The outcome in patients successfully operated upon was satisfactory, with benign residual lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970251 TI - [Characteristics of isolated cardiomyocytes in concentric and eccentric hypertrophy of the human heart ventricle]. AB - Alkaline dissociation of cardiomyocytes was used for investigation of concentric and eccentric myocardial hypertrophy in 8 deceased patients with carions ventricular myocardial mass. For both ventricles and forms of hypertrophy the length, diameter and volume of the cells were found increased, the increase being proportional for the left ventricle (except the volume) and gross for linear enlargement of the right ventricle in eccentric hypertrophy. Absolute number of cardiomyocytes reached maximum in concentric left ventricular hypertrophy showing the same values with normal-mass ventricles in eccentric hypertrophy. PMID- 2970250 TI - Back pain: treatment and prevention in a community hospital. AB - Because back pain is a widespread and costly condition that tends to recur, treatment must focus on both the amelioration of acute symptoms and prevention over the long term. This paper reports a longitudinal evaluation of a program from a community hospital that emphasizes both these aspects. One hundred twenty patients routinely admitted to this program were randomly assigned to treatment and control groups. These groups were assessed for differences in demonstrated physical strength, mobility, body mechanics, and self-care knowledge, and in levels of self-reported exercise, anxiety, and pain. There were significant immediate gains on physical measures of fitness and in observed body mechanics; patients also reported significant gains in physical capabilities at home and in leisure activities. Self-care knowledge also improved. When assessed one year later, original gains in physical strength and mobility were being maintained, and self-reported physical capabilities also remained high. Although demonstrated knowledge of correct body mechanics declined over this period, it was still significantly greater than before the program. In light of these results, we believe that outpatient programs like the one reported here merit careful consideration in an era of concern about rising costs for primary health care. PMID- 2970252 TI - DDAVP and EACA used for minor oral surgery in von Willebrand disease. PMID- 2970254 TI - Cardiac pacemakers and dental equipment. PMID- 2970253 TI - Clinical evaluation of two materials for the bonding of composite resins to dentine: one-year results. PMID- 2970256 TI - BDA replies to AIDS claims. PMID- 2970255 TI - Prophylaxis for infective endocarditis. PMID- 2970257 TI - Isolation and expression of cDNA coding for a new member of the phospholipase A2 inhibitor family. AB - The placental protein PP41,2 was shown to have thromboplastin-inhibitor activity. We used partial amino acid sequence information from PP4 cyanogen bromide fragments to design oligonucleotide probes for the screening of a human placental cDNA library. In addition to the PP4 cDNA we isolated a cDNA coding for a protein with considerable homology which we subsequently termed PP4-X. PP4 and PP4-X belong to the phospholipase A2 inhibitor family, as judged by their homology to lipocortin I and calpactin I3. The full-length PP4-X cDNA encodes a protein of 321 amino acid residues including a fourfold repeat structure. Northern blot analysis using the PP4-X cDNA reveals two hybridizing RNA species of approximately 1400 nucleotides and 2500 nucleotides, respectively. The shorter one could well represent the PP4-X transcript which is in good agreement with the isolated cDNA insert of 1326 nucleotides. Expression of the PP4-X coding sequence in E. coli resulted in the appearance of a protein which crossreacts with antibodies raised against PP4. PMID- 2970258 TI - Reconstituted U1 small nuclear ribonucleoprotein complex restores 5' splice site cleavage activity. AB - Functional reconstitution of U1 small nuclear ribonucleoprotein particle (U1 snRNP) was performed using in vitro transcribed U1 snRNA. Hela cell nuclear extract was depleted of its constituent snRNPs by centrifugation at 100,000 X g. The supernatant was devoid of snRNAs and lacked cleavage activity in splicing reactions using in vitro transcribed beta-globin pre-mRNA as substrate. The resulting pellet which contained the snRNAs, retained 5' splice site cleavage activity in a similar splicing reaction. Supplementation of the inactive supernatant fraction with in vitro transcribed U1 snRNA, partially restored 5' splice site cleavage activity thereby demonstrating the specific requirement of U1 snRNP in the initial stage of pre-mRNA splicing. PMID- 2970259 TI - Insulin-like growth factors are mitogens for rat pheochromocytoma PC 12 cells. AB - We have addressed the issue of a mitogenic effect of insulin-like growth factors IGF-I and IGF-II on the PC 12 line of rat pheochromocytoma cells. The proliferation of PC 12 cells cultured in serum-free medium is stimulated threefold by IGF-I and IGF-II with significantly higher potency than epidermal growth factor, whereas platelet-derived growth factor, nerve growth factor, growth hormone and bombesin are inactive. Two types of IGF receptor are present in PC 12 cells and the dose-response curves suggest that the mitogenic responses to IGF's are mediated by the IGF-I receptor. These results suggest that IGF-I and IGF-II act as mitogens on pluripotent chromaffin cells in the development of the peripheral nervous system and adrenal medulla as well as in promotion of in vivo growth of neural crest-derived tumors. PMID- 2970261 TI - Increase in the actin-activated Mg2+-ATPase activity of smooth muscle myosin by increasing myosin concentration. AB - The actin-activated Mg2+-ATPase activity of smooth muscle myosin was measured in 85 mM KCl, 6 mM MgCl2 in the absence of tropomyosin. The activity was dependent on myosin concentration. Vmax increased as myosin concentration was increased, while the Ka (the apparent dissociation constant for actin) remained the same. The extent of filament formation was also correlated with myosin concentration and most of the myosin monomers existed in 10S conformation. These results suggest that myosin concentration influences the actin-activated Mg2+-ATPase activity by changing the 10S-6S-filaments equilibrium. PMID- 2970260 TI - Insulin-like growth factors in sheep thyroid cells: action, receptors and production. AB - Sheep thyroid cells cultured in serum-free medium were used to study the biologic activity, binding, and production of the insulin-like growth factors (IGFs). IGF I, IGF-II, and insulin stimulated thyroid cell division. Abundant, specific IGF receptors on sheep thyroid cell membranes were identified by binding displacement studies. Maximal specific binding of [125I]-labeled IGF-I and IGF-II to 25 micrograms of membrane protein averaged 21% and 27% respectively. The presence of type I and type II IGF receptors was confirmed by polyacrylamide gel electrophoresis of [125I]IGFs covalently cross-linked to cell membranes. Under reducing conditions, [125I]IGF-I bound to a moiety of approximate Mr = 135,000 and [125I]IGF-II to a moiety of approximate Mr = 260,000. Cross-linking of [125I]IGF-I to medium conditioned by thyroid cells indicated the presence of four IGF binding proteins with apparent Mr = 34,000, 26,000, 19,000 and 14,000. Thyroid cells also secreted IGF-I and II into the medium. IGF synthesis was enhanced consistently by recombinant growth hormone. These data indicate that sheep thyroid cells are a site for IGF action, binding, and production and provide further evidence that IGFs may modulate thyroid gland growth in an autocrine or paracrine manner. PMID- 2970262 TI - Evidence for 5-HT1A binding sites in chick embryo brain and discrimination by 5 methoxytryptamine. AB - The displacement characteristics of [3H]5-hydroxytryptamine (5-HT1) binding by several serotonergic ligands were studied in the chick embryo brain. Although most of ligands tested displaced [3H]5-HT in a manner suggestive of a single site, displacement curves for (+/-)-8-hydroxy-dipropylaminotetralin (8-OH-DPAT), 5-methoxytryptamine, 5-methyltryptamine and 5-methoxy-N,N-dimethyltryptamine displayed non-unity Hill plots, suggesting multiple site interactions. However, if spiperone (2 microM) was included in the assays, the Hill coefficients of these compounds were all similarly increased toward unity, suggesting that 8-OH DPAT as well as 5-methoxy and 5-methyl substituted tryptamines, have a high affinity for and can discriminate 5-HT1A binding sites in the chick embryo brain. PMID- 2970263 TI - Sex hormone status and osteoporosis in postmenopausal women with rheumatoid arthritis. AB - Sex hormones have important effects on bone, especially in postmenopausal women. These hormones may be of particular significance in patients with rheumatoid arthritis (RA), who have a high frequency of osteoporosis. To examine this, we measured estrogen and androgen concentrations and bone mineral density (BMD) in 49 postmenopausal women with RA and 49 normal postmenopausal women. Compared with the controls, postmenopausal RA patients had significantly reduced levels of estrone (median 18 pmoles/liter versus 49; P less than 0.001), dehydroepiandosterone sulfate (DHEAS) (median 0.3 mumoles/liter versus 2.0; P less than 0.001), testosterone (median 0.6 nmoles/liter versus 0.95; P less than 0.001), and femoral BMD (mean 0.72 gm/cm2 versus 0.80; P less than 0.002). Prednisolone therapy in 22 patients (mean dosage 8 mg/day) was associated with reductions in estrone and testosterone levels; however, DHEAS and femoral BMD were also decreased in RA patients who were not receiving corticosteroids. Reduced DHEAS levels in postmenopausal women with RA may increase their risk of osteoporosis. PMID- 2970264 TI - Traumatic injuries among medical center employees. PMID- 2970265 TI - [Special considerations in the dental management of patients with renal transplants]. PMID- 2970266 TI - [Periodontitis in the diabetic]. PMID- 2970267 TI - Suppression of the number of clonally expanding T cells by CD8+ cells as demonstrated by murine T-cell colony formation. AB - Clonally developing murine T cells in the form of T-cell colonies (TCC) in methylcellulose were used to investigate the interaction between polyclonally activated T cells. Only CD8+ T cells proliferate in the methylcellulose and form TCC after stimulation with PHA and IL-2. When the number of developing TCC was counted as a measure of developing clones, it was found that increased cell numbers in the cultures led to decreased percentages of TCC (Number of TCC per 100 seeded cells). This was found already at very low cell concentrations: 40 cells per ml culture, and was maintained at least up to 10,000 cells per culture. Cell sorting (FACS) of cells showed that the suppression of developing clones was mediated via a non-adherent, Thy-1+, CD8+ cell, present in lymph nodes, spleen and the thymic medulla. Such seemingly non-specific suppressor cells may be considered in the network regulation of the functionally mature T-cell population. PMID- 2970268 TI - Attendance patterns for dental treatment of inoculation risk patients. PMID- 2970269 TI - Increase in plasma concentrations of cardiodilatin (amino terminal pro-atrial natriuretic peptide) in cardiac failure and during recumbency. AB - Plasma concentrations of cardiodilatin, the peptide sequence at the amino terminal of the pro-atrial natriuretic peptide, in 17 normal subjects ranged from 59 to 202 (mean 118 (SEM) (9] pmol/l. Recumbency increased the mean (SEM) concentration to 160 (13) pmol/l. The plasma concentration of cardiodilatin in 24 patients with congestive cardiac failure was much higher (964 (175) pmol/l) than in the normal subjects. It was highest in those with heart failure in New York Heart Association functional classes III and IV and the concentration correlated both with atrial natriuretic peptide concentrations and left ventricular ejection fraction. Concentrations rose during induced tachycardia in three patients tested. Chromatography showed a single clean peak of plasma cardiodilatin immunoreactivity. It seems that cardiodilatin is a second circulating cardiac peptide that is jointly released with atrial natriuretic peptide by common stimuli. Other workers have reported that, like atrial natriuretic peptide, three partial cardiodilatin sequences can stimulate renal particulate guanylate cyclase and increase cyclic guanosine monophosphate. The simultaneous release of cardiodilatin in higher circulating concentrations than atrial natriuretic peptide may be relevant to the finding that appropriate concentrations of exogenous atrial natiuretic peptide alone do not produce the full renal effects associated with endogenous peptide release. PMID- 2970270 TI - Anomalous origin of the left anterior descending coronary artery from the pulmonary trunk: recognition in life and successful surgical treatment. AB - An anomalous origin of the left anterior descending coronary artery from the pulmonary trunk with the right and left circumflex arteries arising from the aorta is very rare and the diagnosis made only at necropsy. An anomalous coronary artery was suspected owing to unexplained cardiomegaly in a 17 month old girl and the anatomy was defined by angiography. The anomalous vessel was successfully reimplanted into the aorta. PMID- 2970272 TI - Back pain and sciatica. PMID- 2970271 TI - Reserpine treatment increases viscosity of fluid in the epididymis of rats. AB - Reserpine treatment in rats induces morphological and functional disturbances in exocrine glands which resemble those produced by cystic fibrosis. The general feature is a decrease in fluid secretion with a rise in mucous concentration and altered electrolyte composition. Chronically reserpinized rats have therefore been used as an animal model for the disease. It is known that cystic fibrosis men are infertile due to obstruction of the epididymal duct with inspissated material, a phenomenon that may be secondary to abnormal electrolyte and water transport in the epididymis. Male rats were treated with reserpine (0.5 mg/kg/day) for 12 to 14 days. At the end of the treatment, epididymal fluids were flushed out from the cauda epididymidis for measurement of spermatocrit, viscosity, total protein concentration, sperm concentration and motility. It was found that reserpine treatment caused a rise in viscosity (by 40%), spermatocrit, sperm concentration, and protein concentration. These changes were observed in the epididymis of rats that had been efferent duct-ligated before reserpine treatment. Despite a rise in viscosity of the fluid bathing the spermatozoa, the viability of the stored spermatozoa was apparently normal. Spermatozoa were able to initiate forward motility when suspended in a sodium-containing medium. Testis fluid secretion measured by weight gain after efferent duct ligation for 16 h was not affected by reserpine treatment. The change in viscosity probably was due to a decrease in fluidity in the epididymis. It is concluded that reserpine treatment in rats produced changes in the exocrine functions of the epididymis similar to those seen in other exocrine glands. PMID- 2970273 TI - Effect of stilboestrol and testosterone on the incorporation of 75selenomethionine by prostatic carcinoma cells. AB - Controversy still exists as to whether oestrogens exert a direct effect on the prostatic cell. Incorporation of 75Selenomethionine (SeM) was used as a measure of protein synthesis by prostatic carcinoma cells in vitro to investigate the action of hormones on prostatic carcinoma cells in tissue culture. Stilboestrol (DES) and stilboestrol diphosphate (Honvan) inhibited protein synthesis in a proportion of patients, while testosterone was stimulatory. A similar effect was noted in cells from patients with benign hyperplasia (BPH). This work confirms that oestrogens have a direct inhibitory effect on prostatic cells at high concentrations which can be attained in patients given intravenous stilboestrol diphosphate. PMID- 2970274 TI - ABC of eyes. General medical disorders and the eye. PMID- 2970275 TI - Which patients should undergo laparoscopy? PMID- 2970276 TI - Quinolinic acid-induced seizures, but not nerve cell death, are associated with extracellular Ca2+ decrease assessed in the hippocampus by brain dialysis. AB - Seizures, neuronal damage and extracellular Ca2+ concentration were studied in rats unilaterally injected in the dorsal hippocampus with quinolinic acid, a brain metabolite with excitotoxic properties. In freely moving animals, in the first 2 h after the injection of a convulsant and neurotoxic dose (156 nmol), quinolinic acid induced a tetrodotoxin-insensitive decrease in the extracellular Ca2+ concentration (nadir 40%) in the injected area, as assessed by brain dialysis coupled to a fluorimetric method for Ca2+ detection. Blockade of quinolinic acid-induced decrements in Ca2+ by 15.6 nmol D-(-)2-amino-7 phosphonoheptanoic acid indicated that this effect was receptor-mediated. Dose response relationships showed a close association between seizure activity (measured by EEG) and extracellular Ca2+ changes in the injected area. Changes in Ca2+ were apparent at the site of injection prior to the onset of focal seizures and they were not found in the homotypic structure where seizures were conducted. Drugs effective in blocking seizures (carbamazepine and flunarizine) prevented the fall in extracellular Ca2+, while drugs without anticonvulsant activity (ethosuximide and nifedipine) did not. Destruction of nerve cells by quinolinic acid was not prevented by treatment with carbamazepine and flunarizine. The results suggest that the fall in extracellular Ca2+ observed in the first 2 h after quinolinic acid, probably reflecting the ion influx into neurons, is involved in triggering focal seizures but is not related to the occurrence of nerve cell death. PMID- 2970277 TI - Quantitative autoradiography of serotonin uptake sites in rat brain using [3H]cyanoimipramine. AB - The binding of [3H]cyanoimipramine to serotonin uptake sites in rat brain slices was studied using quantitative autoradiography. Binding was of high affinity and was to a single class of binding site. This is in contrast to results previously obtained by others with [3H]imipramine where two binding sites were observed. The sites labeled by [3H]cyanoimipramine had properties consistent with this ligand labeling serotonin uptake sites, as: (1) binding is displaced by drugs which are potent inhibitors of serotonin uptake but not by drugs which are weak inhibitors of uptake; (2) binding is dependent on the presence of sodium ions as is the uptake of serotonin; (3) binding is almost completely eliminated in the brains of rats lesioned by the serotonin neurotoxin 5,7-dihydroxytryptamine; (4) the distribution of binding sites throughout the rat brain is highly correlated with that found previously for [3H]indalpine, a potent serotonin uptake inhibitor, and for [3H]imipramine. The properties of binding of [3H]cyanoimipramine make it an ideal ligand for the quantitative autoradiography of serotonin uptake sites. PMID- 2970278 TI - Heterogeneous effects of serotonin in the dorsal horn of rat: the involvement of 5-HT1 receptor subtypes. AB - The effect of ionophoretically applied serotonin (5-HT) was tested on cutaneous sensory responses of multireceptive dorsal horn neurones in the anaesthetized rat. Three types of 5-HT action were discerned: selective inhibition of nociceptive responses (10/18 cells), non-selective inhibition of responses to both noxious and innocuous stimuli as well as to excitatory amino acids (4/18 cells) and non-selective excitation of evoked responses (1/18 cells). A few cells (3/18) were unaffected by 5-HT. The use of agonists, shown to discriminate between subtypes of 5-HT1 receptor revealed that a 5-HT1A receptor agonist mimicked the non-selective effects of 5-HT, whereas a 5-HT1B receptor agonist mimicked the selective antinociceptive effects of 5-HT. A 5-HT2 receptor agonist, in contrast, was without effect. Both the selective and the non-selective effects were reversed by a 5-HT1 receptor antagonist, but not a 5-HT2 antagonist. PMID- 2970279 TI - Insulin and insulin-like growth factor 1 receptors during postnatal development of rat brain. AB - The binding of insulin and insulin-like growth factor 1 (IGF1) to high-affinity sites in the brain of rats aged 2-37 days was studied. Specific binding of insulin and IGF1 was assessed using tracer concentrations of 125I-insulin or 125I IGF1. Sites for insulin and IGF1 were distinguished in these conditions as shown by competition experiments. The Kd were 3.6 nM (insulin) and 2.0 nM (IGF1). These values did not change significantly over the age range studied. The numbers of high-affinity binding sites for insulin and IGF1 were similar in adult animals. IGF1 binding was higher than the insulin binding in 2-day-old animals. The binding capacity for both insulin and IGF1 decreased from birth to age 15 and days remained stable thereafter. Tyrosine kinase activity, which is associated with these receptors, was measured using the artificial substrate poly (Glu, Tyr). It decreased over the first 15 days of life and remained stable thereafter. Autophosphorylation of the receptors confirmed this result. This decrease appears to be due to changes in the numbers of the two types of receptors, and is probably a reflection mainly of the variation in the number of IGF1 receptors. Similar results for insulin and IGF1 binding as well as tyrosine kinase activity were obtained with hypothyroid rats. PMID- 2970280 TI - Evidence for vasopressin V1 receptors of rostrodiencephalic neurons: iontophoretic studies in the in vivo rat. Responses to oxytocin and to angiotensin. AB - Extracellular recordings were obtained in anaesthetized rats from single neurons located in various structures around the rostral end of the third ventricle, known to harbour integrative neurons sensing deficiencies in and originating corrective responses for water-electrolyte balance. Once arginine vasopressin (AVP) responsive neurons were located, a selective antidiuretic agonist (binding to V2 receptors) and either V1 (pressor response related) or V2 (antidiuretic) antagonists were iontophoretically applied. Neurons in this region did not respond to the V2 agonist and only the V1 antagonist was able to block the response to AVP. It is assessed that the investigated region has neurons equipped only with receptors of the V1 type. Interestingly, a number of these neurons also responded to angiotensin II (AII), oxytocin and to blood pressure changes. The integrative neuronal population of parasagittal rostrodiencephalic neurons seem therefore to sense indices of haemodynamic changes including their neuro-hormonal signals within the brain such as AII and AVP which bind to V1 (pressor response related) receptors. PMID- 2970281 TI - [Characterization, in the guinea pig, of a hepatic cytosol protein binding dehydroepiandrosterone sulfate and estrone sulfate]. AB - A protein which binds dehydroepiandrosterone sulfate and estrone sulfate was detected in the cytosolic fraction of female Guinea-pig liver. It is characterized by a molecular mass of 14,400 Da, its affinity for DHEA sulfate (KD = 8.8 microM) and estrone sulfate (KD = 8.5 microM), and its lack of affinity for free steroids such as dehydroepiandrosterone or estrone. It is eluted by gel filtration on Sephadex G-50 simultaneously with the inhibitor of microsomal DHEA sulfatase recently described by some of us. This protein could be implicated in the intracellular transport or in the metabolism of sulfated steroids. PMID- 2970282 TI - [Effect of the single intravenous injection of ethanol on exocrine pancreatic secretion in conscious rats]. AB - The studies on the effect of an intravenous infusion of ethanol on the exocrine pancreatic secretion of the rat are contradictory: weak ethanol quantities (less than 2 g/kg) being inefficient, inhibiting or stimulating pancreatic secretion according to different authors. The purpose of this work was to explain these discrepancies. Our results show that: 1) If the secretion of the rats is greater than or equal to 200 microliters/30 min (good general conditions) ethanol inhibits the protein secretion stimulated by either exogenous secretion or endogenous release of CCK (interruption of the flow of pancreatic juice into the duodenum) but does not modify non stimulated secretion (pancreatic juice reintroduced into the duodenum). 2) The differences observed in the literature are probably due to the use of rats in bad general conditions. It is recommended to use only rats, the secretion of which is greater than or equal to 200 microliter PMID- 2970283 TI - [Effects of the addition of carbon dioxide on manifestations of acute hypoxia in rats]. AB - In pentobarbitalized rats, hypoxia induced by inhalation of O2 8%-N2 92%, produces a transient hyperventilation which is followed by a respiratory depression and an apnea. A cardiovascular collapse is then observed. Correction of the hypocapnia depending on the initial hyperventilation, by inhalation of a gas mixture containing 4% CO2 maintains the hyperventilation and suppresses the cardiovascular collapse. Carbon dioxide activity is both a direct one by stimulation of respiratory centers and an indirect one by increasing the sensitivity of the peripheral arterial chemoreceptors to hypoxia. Four percent carbon dioxide just compensating hypocapnia are sufficient to prevent apnea and vascular collapse. The increase of this concentration up to hypercapnia complicates the interpretation of the results by addition of hypoxic and hypercapnic effects. PMID- 2970284 TI - [In vitro differentiation of inflammatory and non-inflammatory states of the synovial fluid by magnetic protonic relaxation]. AB - The differentiation between inflammatory and non inflammatory states has been performed using Nuclear Magnetic Resonance (NMR) in vitro by measuring relaxation times T1 and T2 in 84 synovials fluids obtained from various rheumatologic diseases. The results show that the T1/T2 ratio is more sensitive to distinguish these two situations rather than the isolated T1 or T2 values. In particular, high values of T1/T2 ratio are found in septic arthritis. PMID- 2970285 TI - [Cytometric analysis of the DNA content of the thymus, spleen and liver in C57BL/6 mice]. AB - Flow cytometric analysis of cell DNA content in the mouse C 57 BL/6 shows better results with fine needle aspiration than with mechanical dissociation of the tissues. There is no difference fresh and deep frozen stored tissues. In the liver, it has been found two cell populations with different DNA content including diploid and tetraploid cell lines. In the spleen and the thymus, only diploid cells are encountered with different cell cycles belonging to the organ. PMID- 2970286 TI - [Paradoxical hyperglucagonemia after oral administration of glucose in diabetic dogs. Impact of the cholinergic nervous system]. AB - Alloxan diabetic dogs with insulin deficiency showed a transient but significant rise in glucagon levels after oral glucose load (1 g/kg). Pretreatment with atropine sulfate (0.2 mg/kg intravenously) totally suppressed this increase. So, the transient paradoxical rise of glucagon level observed in diabetic dogs after glucose intake is under cholinergic control. PMID- 2970287 TI - [Immunohistological study of the expression of glucagon in dorsal pancreatic endoderm in the early chick embryo]. AB - An immunohistological study demonstrated that glucagon first appears in the dorsal pancreatic endoderm of the chick embryo at stage 16 of Hamburger and Hamilton during normal development. It was also shown that the self differentiation potency of the isolated dorsal endoderm to express glucagon in vitro in the absence of adjoining tissues appears at stage 11. PMID- 2970288 TI - Computed tomography in the diagnosis of Huntington's disease. PMID- 2970289 TI - Compensatory adaptation of the heart to chronic rate overload: increase in calcium transport ATPase activity of myocardial sarcoplasmic reticulum. AB - This study was undertaken to test the hypothesis that a compensatory response of the heart to a chronic and continuous, metabolic and heart rate overload was an increase in the calcium sequestering activity of the myocardial sarcoplasmic reticulum. Calcium sequestering activity was estimated by determination of the calcium-dependent ATPase (Ca2+-ATPase) activity of isolated microsomes. Chronic rate overload was modelled by comparing: dysthyroid and control rats; control swine and swine with implanted cardiac pacemakers set at 180 beats/min; and different species of mammals with widely different heart rates. The myocardial sarcoplasmic reticulum Ca2+-ATPase pump activity was significantly increased by 39% for hyperthyroid rats compared to control rats and by 87% for control rats compared to thyroidectomized rats; by 63% for paced swine compared to control swine; and by 43% for rats compared to guinea pigs, by 140% for guinea pigs compared to dogs and by 120% for dogs compared to cows. These data indicate that calcium sequestering activity of myocardial sarcoplasmic reticulum increases in equivalent proportion to the chronotropic demand and that heart rate is a hemodynamic correlate of the sarcoplasmic reticulum Ca2+-ATPase activity. PMID- 2970290 TI - Efficacy of delayed percutaneous transluminal coronary angioplasty after intravenous use of streptokinase in myocardial infarction. AB - Between June 1984 and December 1986, 35 patients with acute myocardial infarction received streptokinase intravenously within 3 hours after the beginning of chest pain and underwent percutaneous transluminal coronary angioplasty (PTCA) either immediately (in 2 cases) or 1 to 19 (mean 4.4) days later (in 33). The rate of successful PTCA was 89%. Reocclusion occurred in one patient. The mean percentage of stenosis decreased from 86% to 11%. The mean trans-stenotic gradient was reduced from 41 to 11 mm Hg. The results suggest that in patients whose condition is stable, PTCA performed a few days after thrombolysis is a valuable alternative to more aggressive treatment with immediate PTCA. PMID- 2970291 TI - Mixing studies during hepatic artery infusion in an in vitro model. AB - A glass model of the hepatic artery network was used to study the effect of infusion rate on the degree of mixing from an end hole catheter placed in the gastroduodenal artery. Red dye solutions were infused at rates ranging from 1 ml/hour to 20 ml/minute. Effluent samples from each of 16 branch arteries were collected and dye concentrations were analyzed by means of a spectrophotometer. Low infusion rates, e.g., up to 5 ml/minute, showed streaming of the dye solutions and a nonhomogeneous dye distribution in the distal branches. At 20 ml/minute, dye distribution was much more uniform. These experiments are designed to simulate intrahepatic infusion of chemotherapeutic drug solutions. Theoretical considerations suggesting a pharmacokinetic advantage of intraarterial delivery implicitly assume uniform distribution of drug solutions to all perfused tissue. The in vitro data in this study suggests that this assumption may not be operative under certain infusion conditions. Slow infusion can lead to streaming and nonuniform distribution of infused drug solutions, which may in part explain the variability in tumor response in different tissue regions and also some observed toxicities, such as bile duct stricturing and fibrosis after intrahepatic infusions. PMID- 2970292 TI - T-cells of multiple myeloma patients triggered by the autologous mixed lymphocyte reaction suppress polyclonal immunoglobulin synthesis. AB - To elucidate the possible role of T-cells of patients with multiple myeloma (MM) in the suppression of polyclonal immunoglobulin synthesis. T-cells with and without prior activation by the autologous mixed lymphocyte reaction (AMLR) were added to normal immunoglobulin (Ig)-secreting cultures. The suppression induced by AMLR-activated T-cells from patients with MM was compared to that induced by AMLR-activated T-cells from apparently normal controls. The addition of 10% unstimulated autologous T-cells from patients with MM resulted in minimal suppression of IgG synthesis (87 +/- 19% of baseline values for patients and 115 +/- 21% for controls, no significant difference). The suppression sharply increased when T-cells were preactivated by AMLR and then added in the same concentration to the IgG-secreting cultures (38 + 12% of baseline values for patients compared to 106 + 14% for controls, P less than 0.05). AMLR cultures were performed in the presence of adherent monocytes and after their depletion. The T-cell suppressor effect on normal IgG synthesis was unchanged after monocyte depletion. T-cells preactivated in the AMLR from patients with MM sharply suppress in vitro polyclonal IgG synthesis, and the activation of these suppressor T-cells is not dependent on the presence of monocytes. PMID- 2970293 TI - Novel dehydroepiandrosterone analogues with enhanced biological activity and reduced side effects in mice and rats. AB - Treatment of laboratory mice and rats with the adrenal steroid, dehydroepiandrosterone (DHEA), produces antiobesity and broad spectrum tumor chemopreventive activity. Certain side effects are associated with DHEA administration which could limit its usefulness as a drug. DHEA can be metabolized into both testosterone and estrone and also increases liver weight and liver catalase activity. We have developed two synthetic steroids, 16 alpha fluoro-5-androstan-17-one and 16 alpha-fluoro-5 alpha-androstan-17-one, which, unlike DHEA, do not stimulate uterine weight in sexually immature female rats or seminal vesicle weight in castrated male rats, nor stimulate liver weight and liver catalase activity in mice. 16 alpha-Fluoro-5-androsten-17-one is also about three times as potent as DHEA as an antiobesity agent and is more active when administered p.o. in inhibiting [3H]-7,12-dimethylbenz(a)-anthracene binding to skin DNA and tetradecanoylphorbol-13-acetate stimulation of epidermal [3H]thymidine incorporation in the mouse, two effects believed to be important in the tumor preventive action of DHEA. 16 alpha-Fluoro-5 alpha-androstan-17-one is as active as 16 alpha-fluoro-5-androsten-17-one in inhibiting [3H]-7,12 dimethylbenz(a)anthracene binding to skin DNA and tetradecanoylphorbol-13-acetate stimulation in epidermal [3H]thymidine incorporation but, on the contrary, is not more active than DHEA as an antiobesity drug. Compounds such as 16 alpha-fluoro-5 androsten-17-one and 16 alpha-fluoro-5 alpha-androstan-17-one, which lack specific side-effects of DHEA treatment and demonstrate greater potency, may have therapeutic application as drugs for humans. PMID- 2970294 TI - Phase I clinical and pharmacokinetic study of trimetrexate using a daily x5 schedule. AB - Trimetrexate (TMQ; NSC 352122) is a potent inhibitor of dihydrofolate reductase with good activity against murine i.p.-implanted B16 melanoma and colon 26 tumors. Preclinical antineoplastic activity, demonstrated schedule dependency, and data suggesting effectiveness against methotrexate-resistant cells prompted a Phase I clinical and pharmacokinetic study of trimetrexate using an i.v. daily x5 schedule. Forty-three good performance status patients were treated with 12 dose levels using daily doses varying from 0.5 to 15 mg/m2/d. Plasma and urine samples were obtained for pharmacokinetic analysis using a high-performance liquid chromatographic method. Myelosuppression was dose limiting and 15 mg/m2/d x5 was the maximum tolerated dose. White blood cell (WBC) and platelet toxicity were noted at doses of 1.6 mg/m2 and above. Median WBC and platelet nadirs occurred on approximately Days 11-12 with recovery by Days 15-18. Nonhematological toxicity included mucositis, nausea and vomiting, stomatitis, diarrhea, and rash. Evidence for antitumor activity was seen in seven patients. Trimetrexate elimination from plasma could be represented as either a bi- or triexponential process. Terminal elimination half-lives were in the range of 5-14 h in patients represented by a triexponential model. Approximately 10-20% of the dose administered was excreted in urine over a 24-h period. The recommended starting dose for patients in Phase II trials using the d x5 i.v. schedule is 8.0 mg/m2/d repeated every 21 days. Dose escalations may be possible depending on the extent of prior therapy and individual tolerance of the drug. PMID- 2970295 TI - Effect of medroxyprogesterone acetate on proliferation and cell cycle kinetics of human mammary carcinoma cells. AB - The effect of medroxyprogesterone acetate (MPA) on breast cancer cell proliferation kinetics was investigated in ten human breast cell lines growing as monolayer cultures. Significant inhibition of growth occurred only in the estrogen receptor-positive, progesterone receptor-positive cell lines, T-47D, MCF 7, ZR 75-1, BT 474, and MDA-MB-361. Among these cell lines sensitivity to MPA varied widely; concentrations required for 20% inhibition of growth ranged from 0.04 nM for T-47D to greater than 100 nM for ZR 75-1 cells. Furthermore, although the most sensitive line, T-47D, had the highest level of PR, sensitivity to MPA was not correlated with PR levels among the responsive cell lines. More detailed studies were undertaken with the T-47D cell line. The growth-inhibitory response was confined to the progestins: MPA, ORG 2058, R5020, and progesterone, while androgens, estrogens, and glucocorticoids were without effect over the same concentration range (0.1-100 nM). MPA-induced growth inhibition was associated with a significant decrease in the proportion of S-phase cells with an accumulation of cells in the G0-G1 phase of the cell cycle. Cells began to accumulate in G0-G1 after 12 h of drug treatment and the effect was maximal by 24 h, i.e., maximal effects were observed during the first cell cycle following drug treatment. By contrast, significant accumulation in G0-G1 required exposure of MCF-7 cells to MPA for at least two cell cycle times, i.e., 48 h and the effect was still increasing at 96 h. Stathmokinetic studies revealed that in both cell lines accumulation in the G0-G1 phase was due to an MPA-induced increase in the G1 transit time. These data indicate that MPA and other progestins have direct growth inhibitory effects on estrogen receptor-positive and progesterone receptor positive human breast cancer cells in vitro and these effects can be accounted for by a decrease in the rate at which cells traverse the G1 phase of the cell cycle. PMID- 2970297 TI - [Hypertrophy and left ventricular function in essential arterial hypertension]. PMID- 2970296 TI - Gene for ovarian granulosa cell tumor susceptibility, Gct, in SWXJ recombinant inbred strains of mice revealed by dehydroepiandrosterone. AB - Spontaneous, malignant ovarian granulosa cell (GC) tumors occur in pubertal SWR and specific SWXJ recombinant inbred strains of mice. Treatment of these mice with dehydroepiandrosterone (DHEA), an adrenal secretory steroid with anticancer actions against spontaneous and carcinogen-induced tumors of different tissues, gave unexpected results. Diet supplemented with 0.4% DHEA (a) induced significantly more GC tumors in spontaneous tumor-susceptible strains (SWR and SWXJ-1, -4, and -9), (b) induced the first GC tumors observed in five previously tumor-free strains (SWXJ-6, -7, -8, -10, and -12), and (c) failed to induce GC tumors in SJL and in the remaining six SWXJ strains (SWXJ-2, -3, -5, -11, -13, and -14). The strain distribution pattern of DHEA-induced GC tumor susceptibility versus resistance was compared with strain distribution patterns for 35 different loci known to distinguish SWR and SJL progenitor strains. A complete match of DHEA-induced GC tumors with pancreas-2 (Pan-2) on mouse chromosome 4 was found. We have named this new locus GC tumor susceptibility (Gct), with the Gcts (susceptible) allele found in SWR and the Gctr (resistant) allele found in SJL mice. The Gct locus is closely linked to pancreas-2, Pan-2, but the order of genes is not yet confirmed. In addition, data from F1 progeny of matings between SWR and selected inbred strains provide suggestive evidence for a second gene controlling GC tumor incidence that we hypothesize involves steroid metabolism. Differences in GC tumor incidence data from reciprocal F1 progeny of matings between SWR and SJL mice reveal a strong maternal effect that may represent yet a third gene. These data support a heritable basis for GC tumorigenesis in the SWR model involving a small number of genes. PMID- 2970298 TI - [Treatment of hirsutism by the long-term administration of the anti-androgen cyproterone acetate]. PMID- 2970299 TI - Transcatheter retrieval of intracoronary detached angioplasty guidewire segment. AB - A transcatheter technique of removing a detached angioplasty guidewire segment from the diagonal branch of the left anterior descending coronary artery is described. A standard 0.014 angioplasty guidewire was modified into a "snare" apparatus for retrieval of the detached wire segment. The retrieval was successfully accomplished without intimal dissection, plaque hemorrhage, or vessel occlusion. PMID- 2970300 TI - Percutaneous double-balloon angioplasty of a stenotic modified Fontan aortic homograft conduit. AB - A 28-year-old woman with pulmonary atresia underwent a modified Fontan procedure that utilized a valved aortic homograft; she developed stenosis of the distal anastomosis between the homograft and the pulmonary artery. Because of the increased risk of reoperation, balloon angioplasty of this stenotic lesion was performed successfully. In the subsequent 11 months she has remained asymptomatic. Balloon angioplasty was a successful alternative to reoperation in this patient. PMID- 2970301 TI - Hugging balloons: coronary angioplasty of oversized vessels with side-by-side balloons. AB - Angioplasty of large (greater than 4.0-mm) saphenous vein grafts using the largest available (4.0-mm) balloon catheters may result in inadequate graft dilatation. To avoid leaving a significant residual stenosis, two balloon catheters can be inflated side by side across the stenosis. Herein, we report two cases where this "hugging balloon" technique was used to achieve wide graft patency. PMID- 2970302 TI - Value of immediate coronary angioplasty following intracoronary thrombolysis and acute myocardial infarction. PMID- 2970303 TI - Mechanism of cos DNA cleavage by bacteriophage lambda terminase: multiple roles of ATP. AB - In the terminus-generating (ter) reaction of phage lambda, the phage enzyme terminase catalyzes the production of staggered nicks within the cohesive-end nicking site (cosN). Although the two nicks are related by a rotational symmetry axis that bisects cosN, the in vitro ter reaction is strikingly asymmetric at the nucleotide level. Nicking of the lambda r strand precedes nicking of the I strand. Furthermore, when the two nicking reactions are uncoupled, they have different nucleotide cofactor requirements. ATP plays critical roles during cos cleavage: First, nicking of both DNA strands is stimulated by the addition of ATP. Second, ATP is required for the correct specificity of r-strand nicking since, in the absence of nucleotide, the r-strand nick is shifted 8 bases to the left. Studies with nonhydrolyzable analogs indicate that ATP hydrolysis is not required for these functions. However, after the two nicks are made, terminase catalyzes a disengagement of the cohered ends in a reaction that requires ATP hydrolysis. PMID- 2970304 TI - Down's syndrome: abnormal neuromuscular junction in tongue of transgenic mice with elevated levels of human Cu/Zn-superoxide dismutase. AB - To investigate the possible involvement of Cu/Zn-superoxide dismutase (CuZnSOD) gene dosage in the neuropathological symptoms of Down's syndrome, we analyzed the tongue muscle of transgenic mice that express elevated levels of human CuZnSOD. The tongue neuromuscular junctions (NMJ) in the transgenic animals exhibited significant pathological changes, namely, withdrawal and destruction of some terminal axons and the development of multiple small terminals. The ratio of terminal axon area to postsynaptic membrane decreased, and secondary folds were often complex and hyperplastic. The morphological changes in the transgenic NMJ were similar to those previously seen in muscles of aging mice and rats as well as in tongue muscle of patients with Down's syndrome. The findings suggest that CuZnSOD gene dosage is involved in the pathological abnormalities of tongue NMJ observed in Down's syndrome patients. PMID- 2970305 TI - NBxFO factor--a novel suppressor factor produced by fusion of neonatal spleen and a nonsecreting myeloma. AB - Spleens from 1-day-old DBA/2J mice were fused to the nonsecreting myeloma, FO. Dialyzed supernates of these cells were found to suppress the antigen-specific proliferative response of cloned helper or alloreactive T cells at a final concentration of less than or equal to 5%. The same supernate-containing factor did not suppress the response to IL-2 of an IL-2-addicted T cell line. The factor was found not to suppress the production of either IL-2 or antibody, following stimulation of spleen cells with LPS. Absorption analysis revealed that the target of the factor was the accessory cell population. Further experiments indicated that the factor blocked the proliferation of thymocytes due to IL-1. Biochemical analysis revealed a molecular weight for the factor of about 90,000 and a pI of approximately 4.5. PMID- 2970306 TI - Characterization of cell surface molecules involved in the recognition of antigen presenting cells by cloned helper T-cell lines. AB - The recognition of antigen-presenting cells (APCs) by T helper (TH) cells occurs in an antigen (Ag)-specific, MHC-restricted manner. Recent evidence, however, suggests that other interaction molecules may also be involved in TH:APC interaction in addition to the T-cell receptor (Ti) and class II or la antigens. We chose, therefore, to examine the role of various interaction molecules (Ia, Ti, L3T4, and LFA-1) in Ag presentation using several TH clones with distinct recognition patterns (self-Ia, self-Ia/Ag, and allogenic Ia). We describe here the use of a rapid clustering assay to study the initial binding events that occur between TH cells and APCs of various types. In all combinations of TH cells and APCs, conjugate formation was both Ag-specific and MHC-restricted. Moreover, with one exception cell clustering was prevented by the addition of monoclonal antibodies (mAb) against either the T-cell receptor or class II MHC molecules. In contrast, mAb to L3T4 and LFA-1 generally failed to inhibit cluster formation even though T-cell proliferation was profoundly inhibited. The relative importance of these interaction molecules in conjugate formation appeared to depend on the APC type as well as on the T-cell clone used. The implications of these findings for the mechanisms of Ag presentation and T-cell activation are discussed. PMID- 2970307 TI - The role of accessory molecules in T-helper activation induced by antigen, lectin, or CD3 antibodies. AB - We examined the role of L3T4 and LFA-1 molecules in T-helper-cell activation, under conditions where the physical stability of T helper-accessory cell interactions was not an issue. T-helper hybridomas were activated by accessory cells coated either with concanavalin A (Con A) or with CD3 antibodies. Activation of the T helper cells was measured by microtubule-organizing center (MTOC) reorientation as an early activation event, and by interleukin-2 (IL-2) production as an indication of a fully matured response. Both parameters were strongly blocked by L3T4 and LFA-1 antibodies in the case of Con A activation. In the case of stimulation with accessory cell-bound CD3 antibody, activation was blocked by LFA-1 but not L3T4 antibody. These results support the notion that L3T4 and LFA-1 molecules play more than a simple adhesion role in T-cell activation. The differential effect of L3T4 antibody in the case of Con A activation vs CD3 activation is consistent with the possibility that L3T4 and the alpha/beta portions of the T-cell receptor must interact during antigen- and lectin-stimulated T-cell activation. PMID- 2970308 TI - Infectious and noninfectious tolerance are blocked by a monoclonal antibody to T suppressor factor. AB - Two forms of hapten-specific unresponsiveness have been demonstrated following intravenous (iv) injection of hapten-conjugated syngeneic spleen cell based on the nature of the antigen-presenting cell (APC): I-J+, I-A- APC have been shown to induce T-suppressor cells (Ts cells) which are demonstrated upon adoptive transfer, while I-J-, I-A+ APC induce a nontransferable tolerance. In this paper we report that a monoclonal antibody specific for T-suppressor effector cells and factors (14-12) can block the Ts cells induced by I-J+, I-A- APCs and the tolerance induced by I-J-, I-A+ APCs. In addition, it sufficiently overcomes suppression such that injection of TNP-spl iv induces immunity rather than suppression. We show that the I-A+, I-J- TNP-spl, which induce nontransferable tolerance upon iv injection, are the cells which induce immunity in 14-12-treated recipients. These results demonstrate that injection of I-J-, I-A+ APC does not lead to clonal deletion and the tolerance induced by the iv injection of both I J+, I-A- and I-J-, I-A+ APC operate via Ts cells. PMID- 2970309 TI - Activation of antigen-specific suppressor T cells by the intravenous injection of soluble blood-stage malarial antigen. AB - The regulation of delayed-type hypersensitivity (DTH) to soluble antigens derived from blood-stage parasites was investigated. DTH responses to soluble blood-stage malarial antigen were induced by subcutaneous (sc) sensitization in the flanks and elicited by ear challenge with the same antigen 6 days later. Adoptive transfer studies revealed that T cells of the L3T4+ phenotype were mediating this response. When a high dose of malarial antigen was injected intravenously (iv) prior to sc sensitization, immunosuppression of DTH resulted. The degree of immunosuppression was dependent on the dose of antigen injected iv and the time at which it was administered prior to sc sensitization. Immunosuppression was antigen-specific and mediated by Lyt-2+ splenic T cells. PMID- 2970310 TI - The endpoints of an inversion in wheat chloroplast DNA are associated with short repeated sequences containing homology to att-lambda. AB - The endpoints of an inversion in wheat chloroplast DNA are shown to be associated with copies of a short repeated sequence. Recombination across the repeats in an inverted configuration may have been responsible for the inversion, although they are currently in a direct orientation owing to a second inversion. The repeated sequence contains an element homologous to the core of the bacteriophage lambda att-site, which can function as such in vivo. PMID- 2970312 TI - Genetic analysis of transformation in a microconidiating strain of Neurospora crassa. AB - We have characterized Neurospora crassa transformants obtained with plasmid pDV1001 bearing the cloned catabolic dehydroquinase (qa-2+) gene (Hughes et al. 1983a) and fluffy 268 host strain producing only uninucleate microconidia allowing to isolate individual transformation products. The percentage of transformed nuclei in the mycelium and their stability were determined by genetic analysis of microconidia produced on selective or non-selective medium. About half of the transformants originating from mycelial spheroplasts were apparently homokaryotic. Catabolic dehydroquinase activity was in agreement with the proportion of transformed nuclei. The DNAs from four transformants analyzed by Southern hybridization showed restriction fragments expected for integration of pDV1001 into genomic DNA by non-homologous recombination. No plasmids could be rescued from the undigested DNAs of the transformants by transformation of E. coli. One transformant, fl268-6, was unstable and generated a high proportion of segregants. Plasmid pDV1001 sequences were absent in their DNA. Colonies originating from microconidia of strain fl268-6 on selective plates often lost the transformed character. These results suggest that instability in this transformant is due to the loss of integrated plasmid sequences during vegetative growth. PMID- 2970311 TI - Sporulation-regulated genes of Saccharomyces cerevisiae. AB - We have characterized 46 hybrid phage which hybridize preferentially to mRNA from sporulating cells. Cross-hybridization experiments demonstrate that 27 distinct SPR (Sporulation regulated) sequences are represented among these phage. The SPR genes can be grouped into three classes: early, middle, and late. The early class shows an accumulation of transcripts soon after transfer to sporulation medium and continues to accumulate RNA throughout sporulation. Transcripts of the middle class increase in level at about the time of DNA synthesis, rise rapidly in abundance until meiosis II, then accumulate more slowly for at least the next 3 h. Late gene transcripts begin to accumulate at about the time of meiosis I, increase 10- to 20-fold in the next 2 h, then remain constant in late sporulating cells. PMID- 2970313 TI - Metal ion-activated acid adenosine triphosphatase from chicken liver lysosomes- purification and enzyme properties. PMID- 2970315 TI - [Study on the inhibitory effect of sodium selenite on hepatocarcinogenesis in ducks]. PMID- 2970314 TI - Effects of streptozotocin and of glucose loading on activities of lysosomal thiol proteinases in rat pancreatic tissue. PMID- 2970316 TI - [Clinical significance of anti-HBc and IgM.anti-HBc]. PMID- 2970317 TI - [Immunochemical localization of gastrin peptide in the CNS and gastric antrum]. PMID- 2970318 TI - [Effect of gossypol (+/-, +, -) on Leydig cells of the rat in vitro]. PMID- 2970319 TI - [Purification of TNF and observations on its biological characteristics]. PMID- 2970320 TI - [Electron microscopic observation of the effect of gossypol on atypical hyperplasia of the endometrium]. PMID- 2970321 TI - [Clastogenicity of gossypol as a male contraceptive in sperm head anomalies in mice]. PMID- 2970322 TI - [ELISA for the detection of anti- [Met].enk antibodies]. PMID- 2970323 TI - [Initial investigation of a colorimetric method for determining ornithine carbamoyl transferase in serum and its clinical significance]. PMID- 2970324 TI - [Radiologic-pathologic correlation of solitary pulmonary mass with reference to the pulmonary artery or vein]. PMID- 2970325 TI - [A case of Aint and its family investigation]. PMID- 2970326 TI - [CT diagnosis of high-density renal cysts in an autosomal dominant polycystic kidney (2 case reports)]. PMID- 2970327 TI - [Cryptosporidiosis]. PMID- 2970328 TI - [A survey of medical exposure dose delivered to examinees during X-ray diagnoses in Beijing]. PMID- 2970329 TI - [Protective effect of anisodamine (654-2) in adrenalectomized rats]. PMID- 2970330 TI - [Antioxidation effect of Rhaponticum uniflorum against hematoporphyrin derivative photooxidation]. PMID- 2970331 TI - When can the infusion period be safely ignored in the estimation of pharmacokinetic parameters of drugs in humans? AB - We describe a simple and rapid method to determine the amount by which the area under the curve (AUC) is underestimated when a drug is given by i.v. infusion, but the infusion period is ignored and the post-infusion data are analysed as if they derive from a bolus injection. Charts are provided that allow the investigator to determine the approximate underestimation for drugs following the one- and two-compartment models, and hence to decide whether it is acceptable to ignore the infusion period in a given case (e.g., underestimation less than 5% or 10%). Equations are also provided that allow the exact underestimation to be calculated, together with the true value of the AUC. PMID- 2970332 TI - Ouabain-insensitive active sodium transport in rat jejunum: evidence from ATPase activities, Na uptake by basolateral membrane vesicles and in vitro transintestinal transport. AB - The basolateral membrane of the jejunal enterocyte of the rat was separated by self-orienting Percoll-gradient centrifugation and further purified from brush border contamination. Pellets were analysed for Mg-, Na- and (Na,K)-ATPase activities. The uptake of 0.02 M NaCl was also followed by the rapid micro filtration technique. Transintestinal transport of fluid and electrolytes, and cell water, Na and K were determined in the in vitro everted and incubated jejunum. There is ouabain-insensitive Na-ATPase in addition to the well-known (Na,K)-ATPase in the basolateral membrane. These are differently inhibited by furosemide and ethacrynate. Na uptake by osmotically active basolateral membrane vesicles is enhanced by ATP and a further enhancement is obtained if there is intravesicular K. The ATP effect is inhibited differently by strophanthidin, furosemide and ethacrynate. In the everted sac preparation, transintestinal transport of Na and fluid still occurs when the Na/K pump is totally inhibited by ouabain. These experimental results suggest that there is also a ouabain insensitive Na pump, different from the Na/K pump, in the basolateral membrane. PMID- 2970333 TI - Diminished cardiac hypertrophy and muscle performance in older compared with younger adult rats with chronic atrioventricular block. AB - The combined effect of advancing age and hemodynamic overload on cardiac muscle function has received little attention. In male, Sprague-Dawley rats, we studied the interaction of chronic atrioventricular heart block induced by transvenous electrocautery for 4-12 months (mean, 7 months) and age at study (12, 19 +/- 0.7, and 24 +/- 0.2 months) on cardiac hypertrophy and muscle function compared with age-matched, sham-operated controls. Hypertrophy was determined by the ratio of heart weight to tibial length. Muscle function was first determined from the mechanical variables of the isometric contraction of an excised, thin, left ventricular trabecular muscle bathed at 29 degrees C under a variety of calcium concentrations and stimulation patterns. Then, in the same muscles after disruption of membranes with Triton X-100, the force-pCa curve of the myofibrils was obtained. No hypertrophy occurred with aging in the control group, but alteration in hypertrophy with age occurred in the block group such that the youngest animals with block had the most hypertrophy (170%) and the oldest animals with block the least hypertrophy (120%). The tension developed by cardiac muscle and the duration of the isometric contraction were not affected by age in the control group but were significantly affected by age in the block group. The young animals with block had a markedly prolonged contraction duration and almost twice the developed tension compared with the older animals with block or with controls. The age-related difference in muscle contraction duration in the block group was associated with, and may have only been secondary to, the age-related difference in the extent of cardiac hypertrophy. For developed tension, the age related difference in the block group could not be explained by differences in the extent of cardiac hypertrophy. Rather, this difference was attributable to both an increased myofibrillar force-generating capacity in the young block and to an impairment in excitation-contraction coupling in the old block. The results show that during long-term block, age exerted not only a significant effect on the extent of cardiac hypertrophy but also an independent effect on the developed tension of cardiac muscle. PMID- 2970334 TI - Cellular basis of wall remodeling in long-term pressure overload-induced right ventricular hypertrophy in rats. AB - To determine the effects of long-term pressure overload on the structural mechanisms implicated in wall remodeling of the right ventricle, a mild pulmonary artery banding was applied to rats approximately 2 months old, and the animals were killed 150 days later. The surgical procedure resulted in a 60% reduction in the cross-sectional area of the constricted vessel and a 52% increase in the weight of the right ventricle. The hypertrophic myocardial response was associated with an elevation in right ventricular systolic pressure (from 33 +/- 11 mm Hg to 71 +/- 12 mm Hg), right ventricular end-diastolic pressure (from 3 +/ 1 mm Hg to 10 +/- 3 mm Hg), and central venous pressure (from 2 +/- 0.2 mm Hg to 10 +/- 3 mm Hg). The 76% increase in wall thickness after pulmonary artery stenosis was the result of a 24% lateral expansion of cardiac muscle cells and a 44% increase in the number of myocytes across the ventricular wall. The intermyocyte distance was also increased by 22%. These cellular adaptations occurred with no alterations in total myocyte length, average sarcomere length, and volume composition of the myocardium. Ventricular wall area was decreased by 14%, which suggests a small reduction in chamber volume. Myocyte growth was accompanied by proportional expansions of mitochondrial and myofibrillar components, so that the ratio of mitochondria to myofibrils in the cytoplasm remained essentially constant. In conclusion, ventricular remodeling in this model of chronic pressure hypertrophy is characterized by increases in cellular diameter and number that would both tend to decrease the magnitude of systolic and diastolic stresses on a per cell basis and thus improve the myocardial response to a prolonged and sustained mechanical load. PMID- 2970335 TI - Doppler color-flow imaging assessment of shunt size in atrial septal defect. AB - Two-dimensional echocardiography and pulsed-Doppler studies have not proved to be reliable methods of assessing left-to-right shunt size in atrial septal defect. Doppler color-flow imaging displays the transatrial jet, providing a new dimension with the potential capability of quantifying left-to-right shunt size. Twenty-three patients with atrial septal defect were studied by color-flow imaging and cardiac catheterization. The defect size measured by two-dimensional echocardiography, the maximal color-flow jet width in the atrial septum, and the maximal color-flow jet area in the right atrium were correlated with cardiac catheterization-derived left-to-right shunt size. Correlation coefficients were 0.57 (p less than 0.01), 0.67 (p less than 0.001), and 0.65 (p less than 0.01), respectively. Atrial septal color-flow jet width distinguished patients with less than a 2:1 left-to-right shunt size ratio (eight patients, jet width less than 15 mm in in all) from patients with greater than a 2:1 left-to-right shunt size ratio (15 patients, jet width greater than 15 mm in all). These results indicate that Doppler color-flow imaging can distinguish left-to-right shunt size in atrial septal defect accurately enough to influence decisions with regard to subsequent patient management. PMID- 2970336 TI - Detection, localization, and quantitation of bioprosthetic mitral valve regurgitation. An in vitro two-dimensional color-Doppler flow-mapping study. AB - The usefulness of two-dimensional color-Doppler flow-imaging (2D Doppler) in the detection, localization, and quantitation of bioprosthetic mitral valve regurgitation is uncertain. Mitral bioprostheses, before and after the creation of transvalvular (n = 33), paravalvular (n = 17), or combined (n = 23) defects, were mounted in a pulsed duplication system (flow rates, 2.5-6.5 l/min; pulse rate, 70 beats/min). An Aloka 880 2D Doppler system (Japan) was used to image the regurgitant jets in the simulated left atrial chamber, analogous to images obtained with transesophageal echocardiography. Jet area was corrected to an estimate of stroke volume: 2D Doppler measurements were divided by [(valve effective orifice area) X (continuous-wave Doppler-determined mean diastolic filling velocity)]/pulse rate. Regurgitant fraction and regurgitant volume were measured by an electromagnetic flow probe. 2D Doppler correctly identified the presence and location of paravalvular regurgitation. In transvalvular and combined transvalvular-paravalvular defects, there were six incorrect interpretations, all having transvalvular regurgitant volumes less than 4 ml/beat. In the presence of transvalvular regurgitation, jet area, length, and width correlated linearly with regurgitant volume (r = 0.82, 0.80, and 0.68, respectively; p less than 0.0001) and regurgitant fraction (r = 0.62, 0.61, and 0.45, respectively; p less than 0.001). Correlations with regurgitant fraction were improved when 2D Doppler measurements were corrected for stroke volume (r = 0.78, 0.79, and 0.67, respectively; p less than 0.0001). Mitral bioprostheses with transvalvular defects were also studied at varying flow rates (3.2-7.5 l/min) and pulse rates (70, 90, and 110 beats/min). The correlation between jet area and regurgitant volume was improved with a second-order polynomial regression (r = 0.93, p less than 0.0001). Our conclusions are that 1) in this in vitro model analogous to transesophageal imaging, 2D Doppler accurately detects and localizes bioprosthetic mitral valve regurgitation; 2) in transvalvular bioprosthetic mitral valve regurgitation, 2D Doppler measurement of jet area has a curvilinear relation with regurgitant volume, and correlation with regurgitant fraction is improved with correction for stroke volume; and 3) in paravalvular bioprosthetic mitral valve regurgitation, correlations between 2D Doppler measurements and regurgitant volumes are weaker, possibly because of jet eccentricity. PMID- 2970337 TI - Influence of balloon size on initial success, acute complications, and restenosis after percutaneous transluminal coronary angioplasty. A prospective randomized study. AB - Restenosis after percutaneous transluminal coronary angioplasty (PTCA) is strongly associated with incomplete initial dilatation. To determine if oversized PTCA balloons would reduce the restenosis rate without increasing the risk of arterial dissection and acute complications, we prospectively randomized 336 patients to receive either smaller or larger balloons. Thirty-four percent of patients had multivessel disease and 18% had multisite dilatation. One hundred sixty-nine patients were randomized to PTCA with a larger balloon and 167 to PTCA with a smaller balloon. Balloon:artery diameter ratios were 1.13 +/- 0.14 in the larger group and 0.93 +/- 0.12 in the smaller group (p less than 0.001). The trial was halted as clinically important differences in acute complications emerged. Emergency bypass graft surgery, usually for the treatment of arterial dissection, was required in 7.1% of patients in the larger balloon group and 3.6% of patients in the smaller balloon group (p = 0.15). Myocardial infarction (Q wave and non-Q wave) complicated 7.7% of procedures in which large balloons were assigned and 3.0% of procedures in which small balloons were assigned (p = 0.056). There were no deaths in either group. The incidence of bypass surgery was 1.7% when the balloon:artery ratio was less than 0.9, 3.1% when the ratio was 0.9 1.1, and 7.8% when it was greater than 1.1. Stepwise logistic regression analysis demonstrated that larger balloon assignment, multiple lesion dilatation, and multivessel coronary artery disease were independent predictors of emergency surgery. Angiographic restudy rates were 50% in the larger group and 60% in the smaller group (p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970338 TI - Potential protective effect of high coronary wedge pressure on left ventricular function after coronary occlusion. AB - To assess the potential of coronary collateral circulation to protect myocardium after occlusion of a coronary vessel, the mean coronary wedge pressure, the angiographic grade of collateral channels, and the left ventricular function were studied in 47 consecutive patients with mechanical recanalization of totally occluded coronary arteries. Coronary wedge pressure measurements were obtained 39 +/- 51 days (range, 2 hours to 361 days) after the presumed time of occlusion. The patients were divided into two groups: 31 with a coronary wedge pressure more than 30 mm Hg (group 1) and 16 with a coronary wedge pressure of or less than 30 mm Hg (group 2). Patients in group 1 had a significantly higher mean global left ventricular ejection fraction than those in group 2 (63 +/- 9% vs. 49 +/- 7%, p less than 0.001). Regional left ventricular function (artery-related area change) was also superior in group 1 compared with group 2 (47 +/- 11% vs. 36 +/- 10%, p less than 0.01). Global left ventricular function was significantly correlated to coronary wedge pressure (r = 0.51, p less than 0.001) but not to the angiographic presence of collaterals. The data suggest that a high coronary wedge pressure is associated with improved left ventricular function after coronary artery occlusion and that coronary wedge pressure more accurately reflects the physiological role of collaterals than their angiographic presence. PMID- 2970339 TI - Demonstration of an imbalance between coronary perfusion and excessive load as a mechanism of ischemia during stress in patients with aortic stenosis. AB - Patients with aortic stenosis are susceptible to myocardial ischemia during hemodynamic stress, which may be caused by two mechanisms. First, vascular abnormalities inherent in myocardial hypertrophy may impair coronary vasodilation, limiting the ability to increase coronary blood flow to meet increased metabolic demands. Second, aortic stenosis itself may cause an imbalance between oxygen supply and demand during hemodynamic stress by decreasing aortic pressure (decreasing coronary perfusion or oxygen supply) and increasing left ventricular pressure (increasing oxygen demand). By decreasing aortic valve gradient without immediately altering ventricular hypertrophy, aortic balloon valvuloplasty offers the opportunity to distinguish these mechanisms. We hypothesized that aortic valvuloplasty would improve the balance between myocardial oxygen supply and demand, especially during isoproterenol infusion. Nine patients undergoing aortic balloon valvuloplasty were assessed at baseline and during isoproterenol infusion (5 +/- 2 micrograms/min, mean +/- SD) before and after valvuloplasty. Valvuloplasty increased myocardial oxygen supply. After valvuloplasty, isoproterenol decreased diastolic pressure time index (DPTI) less and increased coronary sinus blood flow more than before valvuloplasty (-630 +/- 367 vs. -292 +/- 224 mm Hg.sec/min, p = 0.02 and 53 +/- 137 vs. 179 +/- 145 ml/min, p = 0.001, respectively). Valvuloplasty also decreased oxygen demand, decreasing systolic pressure time index (SPTI) from 4,135 +/- 511 to 3,021 +/- 492 mm Hg.sec/min (p = 0.0002). Valvuloplasty improved the balance between myocardial oxygen supply and demand, increasing baseline DPTI:SPTI, decreasing aortocoronary sinus oxygen content difference (0.51 +/- 0.15 to 0.68 +/- 0.14, p = 0.005 and 96 +/- 14 to 78 +/- 15 ml O2/l, p = 0.002, respectively), and decreasing myocardial lactate production during isoproterenol infusion (mean lactate extraction fraction, -0.26 +/- 0.40 to 0.14 +/- 0.17; p = 0.01). We conclude that aortic valvuloplasty improves the balance between myocardial oxygen supply and demand during hemodynamic stress induced by isoproterenol infusion. We speculate that the clinical improvement, which often occurs in these patients after valvuloplasty despite persistence of hemodynamically "critical" aortic stenosis, is in part attributable to immediate improvement in the myocardial oxygen supply:demand ratio. PMID- 2970340 TI - Intra-arterial stenting in the atherosclerotic rabbit. AB - The major problem associated with percutaneous transluminal coronary angioplasty is recurrence of the stenotic lesion. Balloon catheter-mounted intracoronary stent devices may reduce restenosis by improving luminal morphology and flow characteristics. This study assessed the effects of a stainless steel wire, interdigitating coil stent on restenosis in the atherosclerotic rabbit model. Fifteen cholesterol-fed rabbits with preexisting iliac arterial lesions induced by balloon deendothelialization were instrumented in one iliac artery with a 2.0 mm diameter stent after balloon dilatation; the contralateral iliac lesion was treated by dilatation only to serve as a control. The animals were given heparin with aspirin (60 mg) and aspirin every 3rd day until death. Arteriography was repeated 4 weeks after stenting, just before death. Tissue sections from stented and control arterial segments were analyzed morphometrically. Stented arteries had a significantly larger luminal diameter at restudy, whether measured by arteriography (1.38 +/- 0.19 vs. 0.94 +/- 0.35 mm, p less than 0.01) or from tissue sections (1.26 +/- 0.18 vs. 0.81 +/- 0.30 mm, p = 0.0001). Wall thickness of the stented segment was slightly, but significantly, less than the control segment (436 +/- 143 vs. 532 +/- 221 microns, p less than 0.05). Scanning electron microscopy of five stented atherosclerotic rabbit aortas revealed regeneration of a nonthrombogenic, confluent, flow-directed endothelium by 4 weeks after placement. Intra-arterial stenting may be of benefit in the prevention of restenosis by the preservation of a larger functional lumen and by a decrease in the neointimal hyperplastic response to arterial injury. PMID- 2970341 TI - Importance of adequate heparin dosage in arterial angioplasty in a porcine model. AB - Acute occlusion after a successful angioplasty is a severe complication that has been reported in 2-12% of patients. Therefore, to determine whether there was a relation between the dosage of heparin and the presence of mural thrombosis, we studied in a pig model the relation between the dosage of heparin and acute platelet-thrombus deposition on the site of arterial injury. We retrospectively analyzed the effect of three heparin regimens on platelet deposition in 32 normal pigs (mean weight, 32.7 kg) that underwent bilateral carotid angioplasty and were sacrificed 90 +/- 26 minutes later. Pigs in protocol 1 (n = 7) received an intravenous bolus injection of 4,000 units heparin 10 minutes before angioplasty. Pigs in protocol 2 (n = 11) received two bolus injections of 4,000 units heparin 40 minutes apart; the angioplasty was performed immediately after the second bolus. Pigs in protocol 3 (n = 14) had an initial 4,000-unit bolus injection of heparin followed immediately by an infusion of 4,000 units/hr; angioplasty was performed 20 minutes after starting the infusion. 111In-labeled platelet deposition on deeply injured (torn into the media) arterial segments were 86.3 +/ 68, 56.2 +/- 56.9, and 37.7 +/- 37.7 X 10(6)/cm2 for protocols 1, 2, and 3, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970342 TI - Platelets, vasoconstriction, and nitroglycerin during arterial wall injury. A new antithrombotic role for an old drug. AB - Endothelial injury in vivo is associated with platelet deposition and a localized platelet-dependent vasoconstrictive response. To assess the influence of nitroglycerin on platelets and vasoconstriction, quantitative 111In-labeled platelet deposition (no platelets x 10(6)/cm2) of the injured segment and the degree of angiographic vasoconstriction (percent diameter narrowing proximal and distal to the dilated segment) produced during injury by balloon angioplasty of the common carotid arteries were studied in heparinized normal pigs that were sacrificed immediately after the procedure. In deeply injured (injury extending through the internal elastic lamina) compared with mildly injured (deendothelialization only) arteries, there was both greater platelet deposition (63.8 vs. 6.9, p = 0.04) and more vasoconstriction (30% vs. 19%, p = 0.02). In the presence of deep arterial wall injury, nitroglycerin given intravenously at a dose sufficient to lower mean arterial pressure by 9 +/- 2% significantly decreased both platelet deposition (16.2 vs. 63.8, p less than 0.008) and the vasoconstrictive response (20 vs. 30%, p less than 0.02) relative to control. However, in the presence of mild arterial wall injury, nitroglycerin decreased vasoconstriction relative to control (10% vs. 19%, p less than 0.01) without causing a significant decrease in the already low level of platelet deposition (5.6 vs. 6.9, respectively; p = NS), suggesting a direct smooth muscle relaxant effect of nitroglycerin. This is the first reported in vivo effectiveness of nitroglycerin in the reduction of platelet deposition after deep arterial injury. PMID- 2970343 TI - Laser angioplasty. What has been learned from experimental studies and clinical trials? PMID- 2970344 TI - Laser angioplasty. Now and in the future. PMID- 2970345 TI - Report of the Joint ISFC/WHO Task Force on Coronary Angioplasty. The International Society and Federation of Cardiology and the World Health Organization. PMID- 2970346 TI - Current prognosis of ischemic mitral regurgitation. Implications for future management. AB - Ischemic mitral regurgitation is a serious and increasingly common clinical disorder, but at present, little is known of the associated prognostic implications, especially in specific therapeutic subgroups. Over a 6.5-year period beginning January 1, 1981, postinfarction mitral regurgitation was demonstrated ventriculographically in 2,343 (19%) of 11,748 patients having significant coronary artery disease defined at cardiac catheterization. Moderate or severe regurgitation was observed in 381 (3%), and among these patients, four treatment groups were defined: Group I (medical, n = 165), Group II (reperfusion, n = 63), Group III (coronary artery bypass only, n = 94), and Group IV (valve replacement or repair in addition to coronary bypass, n = 59). Multivariable regression analysis of survival data in the overall population and in specific treatment groups was performed with the Cox proportional hazards model. Defined and undefined selection biases precluded formal quantitative survival comparisons among some treatment groups; however, unadjusted and adjusted survival analysis for each group revealed several interesting concepts. First, increasing severity of mitral regurgitation had a progressively negative impact on survival prognosis regardless of treatment. Congestive heart failure, the number of associated disorders, acute presentation requiring cardiac care unit admission, diminished ejection fraction, increasing coronary obstruction, and advanced age all worsened prognosis (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970348 TI - A six-year evolution of percutaneous transluminal coronary angioplasty. The Cleveland Clinic experience, 1981-1986. PMID- 2970347 TI - Derivation of neointima in vascular grafts. AB - Vascular prostheses woven from absorbable lactide/glycolide copolymers are replaced by myofibroblast-laden tissue conduits lined by endothelium. When these prostheses are implanted in arteries of animal models, the absorption-replacement reactions are inhibited by Dacron. To determine whether these reactions resulted primarily from transanastomotic pannus ingrowth, prostheses (30 mm x 4 mm) were constructed of three 10-mm long segments with Dacron segments on both ends and a middle polyglactin 910 (PG910) segment. The prostheses were implanted in the aortas of 15 adult New Zealand White rabbits. Resultant prosthesis/tissue complexes were harvested in triplicate at 2 weeks and at 1, 2, 3, and 4 months after implantation. Explants were photographed and sectioned for light microscopy and for scanning and transmission electron microscopy, which showed 100% patency with no aneurysms or stenoses. Inner-capsule thickness in the PG910 segments increased only during the interval from 2 weeks to 2 months; this thickness was statistically greater than either Dacron segment at 1 and 2 months (p less than or equal to 0.004 and p less than or equal to 0.0001, respectively). Proximal and distal Dacron segments did not differ from each other in thickness. Inner capsules of PG910 segments at 1 month were composed predominantly of myofibroblasts, whereas inner capsules of Dacron segments were composed of fibrin coagulum beyond the initial 2 mm of endothelial pannus ingrowth. These data suggest that transanastomotic pannus ingrowth is not the primary source of cells replacing absorbable vascular prostheses. PMID- 2970349 TI - Advantage of desogestrel containing pill in oral contraception: influence on blood lipids and LCAT activity. AB - In the present study the effects of a combined oral contraceptive preparation containing 0.150 mg desogestrel and 0.030 mg ethinylestradiol on lipid metabolism were investigated. In particular, we observed significant increase in HDL cholesterol and apolipoprotein-A-I (apo A-I) and B (apo B). Triglycerides were not significantly modified. The cholesterol esterifying enzyme LCAT, assayed under "maximal" conditions against an exogenous substrate, was significantly decreased despite an increase in the physiological stimulator apo A-I. No changes were observed in the anti-atherogenic indexes apo A-I/apo B and HDL cholesterol/LDL-cholesterol. Thus, it appears that this combined oral contraceptive has the promising ability to increase the anti-atherogenic HDL cholesterol particle without altering the atherogenic LDL-cholesterol. PMID- 2970350 TI - Short term treatment with ketoconazole: effects on gonadal and adrenal steroidogenesis in women. AB - Ketoconazole, an imidazole derivative, is a large spectrum antifungal agent. The drug is known to cause a decrease in plasma androgens and adrenal steroids in normal men; it is also an active drug in the treatment of malignant tumors of the prostate. To examine the antiandrogenic action of this drug in women, we measured several gonadal and adrenal steroids in 21 normally menstruating women before and after receiving oral ketoconazole (200 mg twice daily) for 5 days. Plasma testosterone (T) decreased from a basal level of 0.35 to 0.25 ng/ml (+/- SEM) (P less than 0.001); dihydrotestosterone (DHT) from a basal level of 190.62 +/- 23.2 to 159.75 +/- 19.43 pg/ml (P less than 0.02); dehydroepiandrosterone sulphate (DHEA-S) from 1.42 +/- 0.44 to 1.15 +/- 0.19 micron/ml (P less than 0.02). Plasma 17 beta-estradiol (E2) decreased from a basal level of 97.42 +/- 29.37 to 54.32 +/- 9.9 pg/ml (P less than 0.05). In contrast, plasma 17-OH-progesterone (17-OHP) levels increased from a basal level of 44.81 +/- 8.21 to 71.81 +/- 15.81 ng/100 ml (P less than 0.05). These results confirm that the ketoconazole blocks the conversion of progestins into androgens. The decrease in the plasma concentration of E2 suggest a direct effect of the ketoconazole on the ovary. It is likely that the effect of the drug, both at the level of the ovaries and of the adrenal gland, is dose-dependent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970351 TI - Production in vitro of antibodies directed against alloantigen-specific recognition sites on T cells and on lymphocytotoxic HLA antibodies. AB - We have examined the mechanism of immunological unresponsiveness in a recipient (P.S.) with a long-term functioning renal allograft. P.S., whose HLA type is A1, A30; B14, B18; DR1, w8; DRw52; DQw1 and in whose serum we had earlier demonstrated the presence of antiidiotypic antibodies, received a kidney from a cadaver donor of HLA type A1, A10, B8 in March, 1970. Peripheral blood B lymphocytes from the patient were transformed with Epstein-Barr virus (EBV), and by the cluster-picking technique a B cell line was propagated with continuous production of antibodies. Antiidiotypic antibodies with two distinct biological functions were demonstrable; one specifically inhibiting the lymphocytotoxic activity of anti-HLA-B8, B5, and DR3 reference typing sera, and the other specifically inhibiting proliferative responses in MLC of the recipient's lymphocytes and of third party cells sharing B14, DR1, DQw1 with the patient against stimulator cells carrying B8, DR3 antigens. Immunodepletion experiments demonstrated that the inhibitory activity was associated with the IgM fraction. Absorption experiments suggested that different antibodies may be responsible for the inhibition of lymphocytotoxic activity of anti-HLA sera and of the proliferative responses in MLC. Antiidiotypic antibodies have been postulated to be important in maintaining allograft tolerance in vivo, thereby enhancing renal allograft survival. The availability of such antibodies in large quantities, produced in vitro, could provide antisera for the immunochemical characterization of specific idiotypic receptors on immunoglobulins and T lymphocytes. PMID- 2970352 TI - Generation of functionally active suppressor cells by haemorrhage and haemorrhagic serum. AB - Mitogen (PHA)-induced proliferation of peripheral blood mononuclear cells (PBMC) was reduced by more than 70% 2 h after the haemorrhage of 30% of blood volume. Experiments using isolated macrophages and lymphocytes showed that post haemorrhage macrophages were functionally normal and that lymphocytes were responsible for the observed haemorrhage-induced depression of proliferative response. Surface marker determinations showed that at least some, if not all, of the haemorrhage-induced suppressor cells are of the OX8+ phenotype. Exposure of PBMCs to serum from bled animals also brought about activation of OX8+ suppressor T cells. These results suggest that the depressed proliferative response of PBMCs induced by haemorrhage or by exposing the cells to haemorrhagic serum (serum from bled animals) is due to the activation of OX8+ suppressor T cells. PMID- 2970353 TI - Lymphocytic infiltration and HLA-DR expression of salivary glands in bone marrow transplant recipients: a prospective study. AB - Lymphocytic infiltration and epithelial HLA-DR expression of lip salivary glands were studied by means of an immunohistoenzymatic staining technique in patients undergoing repeated lip salivary gland biopsies before, and 12, 26, 52 and 104 weeks after bone marrow transplantation (BMT). Within 12 weeks of transplantation, lymphocytes, mainly of the anti-Leu3a+ T 'helper' phenotype, were seen infiltrating the salivary glands of all the patients, reaching a maximum between 26 and 52 weeks. Epithelial HLA-DR expression, present at the 12th week after BMT, was seen close to the lymphocytic infiltrates in all the specimens. Two years after BMT, lymphocytic infiltrates and epithelial HLA-DR expression were still noted in about half of the specimens but not seen in the remaining ones. No correlations were found between immunohistopathology and earlier or persistent chronic graft-versus-host disease or immunosuppressive treatment. The significance of the findings as well as their resemblance to idiopathic connective tissue diseases, notably Sjogren's syndrome, are discussed. PMID- 2970355 TI - Two distinct subsets of patients with systemic lupus erythematosus. AB - This study was undertaken to examine the levels and function of peripheral blood immunoregulatory T cell subpopulations in systemic lupus erythematosus (SLE). T cell subpopulations can be distinguished by the T cell differentiation antigens CD4 (recognized by the monoclonal antibodies OKT4 or Leu3) and CD8 (recognized by the monoclonal antibodies OKT8 or Leu2). All SLE patients tested had normal percentages of CD8 cells in their peripheral blood. The SLE patients, however, fell into two groups based on their CD4 cell numbers. Fifty-five percent of the SLE patients had normal levels of CD4 cells (Group A) and therefore normal CD4/CD8 cell ratios, whereas 45% of the SLE patient population had markedly depressed CD4 cell levels (Group B) and significantly low CD4/CD8 cell ratios. T cells from normal donors and SLE patients were further examined for their ability to stimulate allogeneic normal B/M phi cells to secrete IgM in the presence of pokeweed mitogen (PWM). Utilizing this assay system two forms of immunosuppression were observed: (1) that mediated by high concentrations of purified CD4 cells and (2) that mediated by CD8 cells. High concentrations of purified CD4 cells, added to a constant number of allogeneic normal B/M phi cells, suppressed PWM-stimulated IgM synthesis. Group B SLE patients, with significantly low CD4 cell numbers, had defective CD4 cell-mediated suppression which was concentration dependent. This result was confirmed in a study using identical twins discordant for SLE. In this case CD4 cells from the SLE twin did not induce immunosuppression at a high concentration of CD4 cells whereas similar concentrations of CD4 cells from the normal twin resulted in suppression. SLE patients (Group A) with normal levels of CD4 cells had normally immunosuppressive CD4 cells. Suppression mediated by CD8 cells was demonstrated by the fact that removal of CD8 cells resulted in enhanced IgM synthesis induced by the remaining CD4 cells. Although all the SLE patients in this study had normal peripheral blood levels of CD8 cells, SLE Group A patients had defective CD8 cell suppression whereas CD8 function appeared to be normal in Group B patients. These results suggest that in SLE patients with depressed CD4 cell numbers (Group B) there is a corresponding defect in CD4 cell function. We demonstrate that in SLE Group B patients, defective suppression is due to a subset of T cells that bear the CD4 antigen. The SLE patient population (Group A) with normal CD4/CD8 ratios and normally functioning CD4 cells, however, appear to have normal CD4 cell mediated suppression but defective CD8 suppressor cell function. PMID- 2970354 TI - The influence of the normal microbial flora on the susceptibility of rats to experimental autoimmune thyroiditis. AB - Female PVG/c strain rats maintained under specific pathogen-free conditions until weaning were found to be significantly less susceptible to the induction of autoimmune thyroiditis by thymectomy and irradiation than conventionally reared rats of the same strain. Other SPF-derived strains had a similarly low susceptibility, thus pointing to an important external factor influencing the induction of autoimmunity by this procedure. It was found that the oral administration of antibiotics followed by fresh, homogenized, intestinal contents obtained from conventionally reared rats to newly weaned SPF PVG/c rats significantly augmented their autoimmune susceptibility. The offspring of female SPF rats which had been treated in late pregnancy with oral antibiotics and conventional intestinal contents were similarly more susceptible than the offspring of non-treated counterparts. It is proposed that the composition of the normal gastrointestinal flora of conventionally reared rats profoundly influences susceptibility to the induction of autoimmune thyroiditis in this particular experimental model, possibly by antigenic cross-reactivity with thyroid tissue. PMID- 2970356 TI - Heterogeneity of lymphokine-activated killer (LAK) populations at the clonal level: both NK and CD3+, CD4-, CD8- clones efficiently mediate tumor cell killing. AB - To investigate at the clonal level the phenotypic and functional properties of interleukin 2 (IL-2) activated killer cells (LAK), recombinant IL-2 activated peripheral blood lymphocytes were cultured under limiting conditions. Among 56 clones that lysed P815 in the presence of phytohemagglutinin (PHA) (22% of total proliferating microcultures) 36 clones lysed also the natural killer (NK) sensitive K562 and the NK-resistant Hu126 glioma cell lines and one clone lysed only the K562 cell line. Several LAK clones were further assayed for both phenotype and functional activity. Of 22 clones, 10 were CD3-, CD4-, CD8-, and expressed the CD16 marker of NK cells; only one clone had the conventional phenotype of cytolytic T cells (CD3+, CD4-, CD8+), while 11 clones were CD3+, CD4 , CD8- and did not express alpha/beta heterodimer of T-cell antigen receptor as identified by WT31 monoclonal antibody. Only one of the latter clones was CD16+. Endogenous production of IL-2 after stimulation with PHA and phorbol myristate acetate was positive in 3/9 CD3- and in 8/8 CD3+, CD4-, CD8- clones. CD3- mediated strong antibody-dependent cellular cytotoxicity, a function exerted also by some CD3+, CD4-, CD8- T-cell clones to a lower extent. CD3+, CD4-, CD8- T-cell clones lysed different major histocompatibility complex unrelated tumor targets; moreover, this lytic activity seems to be CD3 dependent. PMID- 2970357 TI - Treatment of patellofemoral instability in Down's syndrome. AB - In a retrospective review, 16 patients with Down's syndrome with the chief complaint of patellofemoral joint dislocations or subluxations were studied. Twenty-six dislocatable or dislocated patellae were seen in these 16 patients. The degree of patellofemoral instability was not correlated with either the ambulatory status or the form of treatment (operative/nonoperative). An increasing degree of patellofemoral instability was correlated strongly with the presence of deformities. Nonoperative treatment either maintained or improved the ambulatory status in 67% of these knees with either fair or good ambulation, whereas 80% of the knees with poor ambulation did not improve. Operative treatment resulted in good ambulatory ability in 86% of the knees with fair or poor preoperative ambulatory status, but did not effectively correct deformities that subsequently resulted in degenerative arthritis. PMID- 2970358 TI - Repair of Achilles tendon ruptures with Dacron vascular graft. AB - A technique has been developed for the repair of Achilles tendon ruptures that allows for early mobilization of the patient. A Dacron vascular graft is woven from distal to proximal and across the site of the rupture in a Bunnell-type fashion. The patients are immobilized in a short-leg cast for two weeks and are then fitted for a posterior fiberglass splint. Seven patients with acute ruptures who were treated with repair with the Dacron graft were followed for a period ranging from ten to 38 months. They were allowed to return to their normal level of activity approximately five months after surgery. There have been no reruptures. This technique also holds particular promise for the late reconstruction of an Achilles tendon rupture as well as for the treatment of partial tears in the severely degenerated tendon. PMID- 2970359 TI - [A case of hereditary chorea with elliptocytosis]. PMID- 2970360 TI - Atrial natriuretic peptide inhibits osmolality-induced arginine vasopressin release in man. AB - 1. Eight normal volunteers were infused with 5% saline (5 g of NaCl/100 ml) at a rate of 0.06 ml min-1 kg-1 for 120 min to increase plasma osmolality and plasma arginine vasopressin. Human atrial natriuretic peptide (alpha-hANP; 100 micrograms) or placebo was given in random order in a double-blind cross-over design for the last 20 min of the saline infusion. 2. Compared with the placebo infusion, atrial natriuretic peptide (ANP) produced a 43% greater sodium excretion and a 34% greater urinary volume in the subsequent hour. 3. Mean plasma immunoreactive ANP did not increase in response to changes in osmolality and rose to a peak of 118 pg/ml during the alpha-hANP infusion. alpha-hANP produced significant suppression of mean plasma arginine vasopressin over the 60 min after the infusions. 4. We conclude that ANP is not released in response to increased osmolality in vivo, and that it inhibits osmolality-induced arginine vasopressin release in man. PMID- 2970361 TI - Effect of a meal containing protein on lithium clearance and plasma immunoreactive atrial natriuretic peptide in man. AB - 1. The effect of meals with a high and low protein content and of the fasting state on renal function and plasma atrial natriuretic peptide was studied in water-loaded normal volunteers. 2. Creatinine clearance increased after the high protein meal, but did not change after the low protein meal or while fasting. Observations of similar increases in urine sodium and potassium excretion and a transient decrease in urine flow after both meals suggest that the protein content of the meal is not an important contributory factor in these responses to feeding. 3. Absolute delivery of sodium and water out of the proximal tubules (assessed by the lithium clearance method) was higher after both meals than while fasting; fractional lithium clearance was higher after the low protein meal than the high protein meal and while fasting. Absolute reabsorption from proximal tubules was increased after only the high protein meal. 4. A transient decrease in the fraction of water delivered to distal nephron segments that appeared in the urine (fractional distal water excretion) was observed after both meals. Fractional distal sodium excretion and absolute distal sodium and water reabsorption increased after both meals. 5. Since plasma atrial natriuretic peptide either decreased (high protein meal) or remained unchanged (low protein meal and fasting), it is unlikely that this hormone is involved in the hyperfiltration after the high protein meal and the natriuresis after both high and low protein meals. PMID- 2970362 TI - Endocrine and renal response to water loading and water restriction in normal man. AB - 1. Nine normal subjects (eight male, one female) on a fixed daily intake of 150 mmol of sodium and 80 mmol of potassium, were randomized to receive either 3 days of 1.0 litre total water intake/24 h (food + fluid) or 4 days of 6.8 litres total water intake/24 h, and were then crossed over after a 3 day control period (2.7 litres water/24 h). 2. During water restriction, urine volume fell from 1.94 litres/24 h to less than 1 litre/24 h by the first day and was 0.77 litre/24 h on the final day. Plasma atrial natriuretic peptide levels were unchanged from baseline despite a large increase in plasma vasopressin and plasma and urine osmolality. Urinary sodium was unaltered throughout, while urinary potassium was increased on the final 2 days of water restriction. 3. During water loading, urine volume increased from 1.85 litres/24 h to 5.44 litres/24 h on the first day and remained at approximately 6 litres/24 h for the final 3 days. Plasma atrial natriuretic peptide showed no change. Plasma vasopressin and plasma and urine osmolality were reduced. Urinary sodium and potassium output were unchanged from baseline. 4. These results suggest that changes in plasma atrial natriuretic peptide are unlikely to be involved in the normal homoeostatic response to changes in water balance in man. PMID- 2970363 TI - Dentists professional and ethical responsibilities for HIV-positive patients and patients with AIDS. PMID- 2970364 TI - Effects of fructose 2,6-bisphosphate and glucose 1,6-bisphosphate on phosphofructokinase from chicken erythrocytes. AB - 1. Phosphofructokinase (EC 2.7.1.11) from chicken erythrocytes is activated by fructose 2,6-bisphosphate, glucose 1,6-bisphosphate and AMP, and it is inhibited by 2,3-bisphosphoglycerate and inositol hexaphosphate. 2. The stimulatory effects produced by the two bisphosphorylated hexoses are additive and the effects produced by fructose 2,6-bisphosphate and by AMP are synergistic. 3. The activatory effect produced by fructose 2,6-bisphosphate is counteracted by fructose 1,6-bisphosphate. 4. The inhibition produced by both 2,3 bisphosphoglycerate and inositol hexaphosphate is released by fructose 2,6 bisphosphate. 5. It is concluded that, like phosphofructokinase from mammalian tissues, the enzyme from chicken erythrocytes can be modulated by the relative concentrations of those metabolites. PMID- 2970365 TI - Special holidays. Special children welcome. PMID- 2970366 TI - Special holidays. Holidays with care. PMID- 2970369 TI - Percutaneous transluminal angioplasty following endarterectomy. AB - Prior to the introduction of percutaneous transluminal angioplasty (PTA), bypass grafting or endarterectomy was the treatment of choice for aortoiliofemoral atherosclerotic occlusive disease. Currently, PTA is a well-established procedure for the treatment of aortoiliofemoral atherosclerotic occlusive disease. PTA is as effective as, and safer than, surgery in these cases. Percutaneous transluminal angioplasty and endarterectomy cause similar trauma to the arterial wall, i.e., intimal denudation, plaque disruption and splitting, and medial disruption, splitting, and overstretching. Both PTA and endarterectomy heal in a similar manner, i.e., neointima formation and scarring. Both PTA and endarterectomy can be repeated. Therefore, PTA can be performed after endarterectomy or vice versa. Several patients with recurrent occlusive disease after endarterectomy have been safely treated with PTA. Our results obtained with PTA in patients who had a prior endarterectomy are comparable to the results obtained in patients who did not have a prior endarterectomy. No complications have been encountered. A previous endarterectomy does not preclude a subsequent PTA, or vice versa, in patients with recurrent occlusive disease. PMID- 2970368 TI - Histological changes in the ovary and uterus of rat after injectable contraceptive therapy. AB - Histological changes were studied in the ovary and uterus of rats receiving different doses of Depo-Provera (DMPA) and Noristerat (NET-EN) for varying duration. The effect of DMPA on ovarian and uterine tissues was strongly progestational. The whole morphological alteration in the ovary after DMPA therapy appeared to be the atresia of the follicular apparatus with degeneration of the growing follicles. Uterine histology reflected that endometrial tissues gradually became inactive and with prolonged treatment at high doses, atrophy of the endometrium was noted. In NET-EN-treated rats, absence of mature follicles and recent corpora lutea reflected the blockade of ovulation. There was no extreme atrophy of the ovary or endometrium as found with DMPA treatment. With higher doses of NET-EN, endometrial growth was arrested. PMID- 2970367 TI - A study of adrenal corticol function and its reserve activity in long-acting injectable contraceptive users. AB - Adrenal function and its reserve capacity were studied in eighteen healthy non lactating women who received depot-medroxyprogesterone acetate (DMPA: 9 subjects) and norethisterone enanthate (NET-EN: 9 subjects) as long-acting injectable contraceptives. Methodological assessments included: (i) morning fasting concentration of serum cortisol; (ii) diurnal variation of adrenal corticol activity as evidenced by cortisol concentration in serum samples taken at 0700 and 1600 hours; (iii) urinary free cortisol excretion; and (iv) cortisol response to insulin-induced hypoglycaemia. None of these assessments was significantly altered by contraceptive administration. This finding indicates that the subject's ability to release cortisol in response to a stressful situation, as well as to normal day-to-day activity, was not impaired by these contraceptive steroids. PMID- 2970370 TI - Long-term preservation of dried phosphofructokinase by sugars and sugar/zinc mixtures. AB - We have demonstrated that sugars and suger/zinc mixtures can be used to preserve the activity of dried phosphofructokinase (PFK) during long-term storage over CaSO4. After 9 weeks in the presence of either 200 mM sucrose or 200 mM trehalose little loss of PFK activity was noted, with almost 60% of the original prefreeze dry activity recovered when samples were rehydrated. Even reducing sugars protected the dried enzyme throughout the entire storage period. Of the sugars tested, 200 mM lactose provided the most stability to PFK; at the end of the dry storage, over 80% of the initial activity was recovered. With either 200 mM maltose or 400 mM glucose, about 40% of the initial activity was recovered at the end of the experiment. With all the sugars tested, the addition of 0.6 mM Zn2+ to sugar/PFK mixtures enhanced the stability of the enzyme, and no long-term adverse effects of the metal ion on enzyme activity were noted. PMID- 2970371 TI - Clinical use of hematoporphyrin derivative and photoradiation therapy in nasopharyngeal carcinoma. PMID- 2970372 TI - A novel hormone candidate. Rat atrium cross-reactive with antibody against pre proANF. PMID- 2970373 TI - Pathogenesis of amyloid formation in Alzheimer's disease, Down's syndrome and scrapie. AB - Paired helical filaments (PHF) are abnormal fibrous structures found in human nerve cells and their processes. Ultrastructural studies of the proto-filaments that make up the PHF revealed that the individual proto-filaments have a different substructure from normal neurofilaments or any other known fibrous profiles. Studies using immunological and biochemical methods suggested that abnormally phosphorylated tau, ubiquitin and neurofilament peptides are part of the PHF. Deposits of amyloid fibres in Alzheimer's disease and senile dementia of the Alzheimer type (AD/SDAT) are found in meningeal and brain vessels, choroid plexus and neuritic plaques. In 1984 Glenner and Wong reported the sequence of a beta-protein isolated from cerebrovascular amyloid. We used the amino acid sequence of the cerebrovascular amyloid protein to synthesize oligonucleotide probes specific for the gene encoding this amyloid protein. Screening of a human brain cDNA library allowed us to isolate a clone which encodes the amyloid peptide. In situ hybridization studies and Southern blot analysis of a DNA sample isolated from a human-mouse hybrid cell line indicated that the corresponding genomic sequences of this cDNA clone are located on human chromosome 21. Using immunochemical and histochemical methods, we have identified the cells associated with the formation of the amyloid fibres. With immunochemical and biochemical methods we and others also showed that the protein constituting amyloid in AD/SDAT is different from amyloid in unconventional slow virus diseases. PMID- 2970374 TI - [Allergic contact dermatitis caused by Asteraceae. Identification of an 8,9 epoxythymol-diester as the contact allergen of Arnica sachalinensis]. AB - A pharmacist developed an allergic contact dermatitis while working with Arnica sachalinensis. Patch testing with sesquiterpene lactones was negative. The contact allergen of A. sachalinensis was identified as 10-acetoxy-8,9-epoxy thymol-isobutyrate. This compound, or structurally related 8,9-epoxythymol diesters, are often found in Asteraceae. In addition to sesquiterpene lactones, epoxythymol-diesters should be considered as contact allergens. Cross reactions between the two groups of compounds are not expected and could not be demonstrated in our patients. PMID- 2970375 TI - Skin disorders and prick and patch test reactivity in Finnish reindeer herders. AB - In addition to an interview, skin tests and clinical examinations were performed on 211 randomly selected reindeer herders from 21 to 77 years of age (mean 45 years). Skin tests consisted of skin prick tests (SPT) with 9 allergens: cat and cow epithelium, dog, horse and reindeer epithelium, house dust mite, birch pollen, meadow grass pollen and mugwort pollen, as well as patch tests with 31 allergens. Of the 211 tested, 19 (9%) had positive SPT reactions to at least one allergen, and 19/173 (11%) showed a positive patch test result. Thirty-six of the 211 (17%) had past or present atopic dermatitis. These findings suggest that the prevalence of immediate and contact allergies and skin diseases is roughly the same as that of other Finns. A positive SPT reaction to birch pollen was encountered less frequently than in a previous Finnish study conducted in southeastern Finland. PMID- 2970376 TI - [Allergies to 8-hydroxyquinoline derivatives]. AB - Evaluation of our standardized test results from 1972 to 1983 indicated Vioform allergies in 1.2% of all cases and Sterosan allergies in 1.1%. Our results are within the spectrum found in the literature for Vioform between 0.3 and 5.1% and for Sterosan between 0.3 and 3.7%. In the course of those 12 years we observed a growing tendency towards Vioform and Sterosan allergies on the basis of annual percentage figures. In the last year of our observation the frequency of Vioform and Sterosan allergies amounted to 1.7% each. We conclude, therefore, that inclusion of these two substances in the standard battery is justified. According to the pertinent literature, Sterosan and Vioform allergies are, on an average, 3 to 4 times more frequent in patients suffering from leg ulcers and/or chronic venous insufficiency than amongst eczema patients. PMID- 2970377 TI - [Group allergies and multiple reactions in patients with allergy to hydroxyquinoline derivatives]. AB - We carried out patch testing with a chinoline block comprising 25 substances in 316 patients with a suspected allergy to chinoline derivatives in the period from February 1969 to December 1983. 110 patients reacted to 2 or more substances. The reactions were totaled and tabulated in a list indicating frequency of sensitisation as decreasing in the order of Cl-F-Br-I halogens. The most frequent reactions observed were with Chlorisept, Sterosan, Vio-form, Chinosol, Ondron, and Broxychinolin, for all of which primary sensitisations are also possible. Concomitant reactions to chinaldol, chinaldine, aminochinaldine and chinoline are considered to be group allergies since contact is hardly possible with these substances in everyday life. Due to the lack of appropriate history, reactions to Resochin and the isoquinoline compounds Papaverine and Eupaverine may also be regarded as group allergies. Accordingly, non-hydroxylized chinoline derivatives are also viewed as group allergies in cases of sensitisation to hydroxy-chinoline derivatives. The group spectrum of hydroxy-chinoline allergy seems to be more far reaching than was assumed. Five cases of a quinine allergy identified by appropriate history and confirmed epicutaneously are presented with a group allergy to various chinoline derivatives, including two cases with quinidine. PMID- 2970378 TI - [Allergic contact eczema caused by dicyanodiamide]. AB - After four years employment in a factory manufacturing flame retardants, a 29 year old man developed eczematous reactions on both hands. Patch tests performed with different agents handled by the patient solely at his place of work were positive for the flame retardant and dicyanodiamide, even in high dilutions. Upon changing jobs, the lesions healed completely. To our knowledge, this is the first recorded instance of a contact allergy due to dicyanodiamide in the pertinent literature. PMID- 2970380 TI - Plaque formation and plaque inhibition. PMID- 2970379 TI - [Which hormone therapy should be used in advanced breast carcinoma in males?]. AB - A 76-year-old man with advanced carcinoma of the breast who had not been orchiectomized underwent sequential hormonal treatment. The tumour progressed during monotherapy with dexamethasone. Objective regression of the tumour followed sequential administration of tamoxifen (twice) and the two aromatase inhibitors, aminoglutethimide and testololactone. In addition, his condition remained stable on two progestogens in high doses, medroxyprogesterone acetate and megestrol acetate. PMID- 2970381 TI - [Sugar substitutes--new aspects]. PMID- 2970382 TI - [Effect of levamisole on the activity of the blood T-lymphocytes and of factors they produce in patients with laryngeal cancer]. AB - The functional activity of T-lymphocytes was studied in 20 donors and in 59 patients with laryngeal tumours, stages II to IV. A considerable decrease in the activity of T-lymphocytes was observed in patients with laryngeal cancer, stages III and IV. Levamisole increased the functional activity of T-lymphocytes in vitro. It was found that spontaneous rosette-formation, PHA-induced lymphocyte proliferation and gamma-interferon production increased while the activity of suppressor T-lymphocytes induced by concanavalin A decreased. PMID- 2970383 TI - Effects of eel calcitonin on plasma beta-endorphin-like immunoreactivity levels in rats. AB - The effect of synthetic eel calcitonin (CT) on the level of plasma beta-endorphin like immunoreactivity (beta-En-LI) and on its content in hypothalamus and anterior pituitary in vivo and on the release of beta-En-LI-from anterior pituitary in vitro was studied. The level of beta-En-LI was increased significantly in a dose-dependent manner after the injection of 1-100 U/kg CT, maximum level being found at 10 min after the injection. After the administration of 50 U/kg CT, the increase was still significant at 40 min. Such an increase was not found in hypophysectomized animals. The content of beta-En-LI in the hypothalamus and anterior pituitary did not change significantly after CT injection. In vitro, the release of beta-En-LI from anterior pituitary was enhanced significantly after CT. It may be concluded that CT acts on the pituitary to increase the release of beta-En-LI which results in increased plasma level. PMID- 2970384 TI - Molecular mechanics calculation of geometries of NAD+ derivatives, modified in the nicotinamide group, in a ternary complex with horse liver alcohol dehydrogenase. AB - The geometry of seven NAD+ analogues bound to horse liver alcohol dehydrogenase (LADH) modified only in their nicotinamide group, have been studied using AMBER molecular mechanics energy-minimization procedures. Starting geometries were taken from X-ray crystallographic data for NAD+/Me2SO/LADH reported by Eklund and co-workers. In this study the NAD+ analogues were encaged by the constituent amino acids of the enzyme within a range of 0.6 nm from the initial NAD+/Me2SO/Zn2+ complex. The calculational method used is able to rationalize individual substituent effects and to evaluate the essential interactions between NAD+ analogue, enzyme, Me2SO and Zn2+ without the necessity of additional X-ray data. The results presented here demonstrate that the reactivity of NAD+ derivatives as reported in literature can be qualitatively related to the position of the pyridine moiety in the active site. PMID- 2970385 TI - Evaluation of modified trypticase soy broth versus supplemented peptone broth in the detection of bacteremia and fungemia. AB - In vitro, 1.2% gelatin counteracts the inhibition of growth of bacterial species by sodium polyanetholsulfonate in blood culture media. Additionally, 1% yeast extract has been used to promote bacterial growth. We compared the performance of supplemented peptone broth and Trypticase soy broth, both of which contained sodium polyanetholsulfonate, gelatin and yeast extract. Trypticase soy broth with gelatin and yeast extract inhibited (p less than 0.001) and delayed growth, especially of gram-positive (p less than 0.01) and gram-negative (p less than 0.005) anaerobic bacteria. Although the recovery of organisms usually inhibited by sodium polyanetholsulfonate was similar in supplemented peptone and Trypticase soy broths, supplemented peptone broth clearly was superior in the recovery of other organisms commonly found in blood cultures. PMID- 2970386 TI - Serological response in patients with chronic asymptomatic human immunodeficiency virus infection. AB - To examine a possible relationship between decreased immune function and serological parameters, such as human immunodeficiency virus (HIV) antigenaemia and the quality and quantity of whole virus antibodies and antibodies against the major core protein p24, we investigated 160 healthy HIV infected individuals (CDC classification II and III). According to the number of T-helper lymphocytes (CD4 cells) these were divided into two groups (CD4 cell counts above or below 500/microliter), which according to the lymphocyte transformation response to pokeweed mitogen (response above or below 20% of control value) were further subdivided into two groups. Both the presence of HIV antigen (p = 0.022) and the absence of p24 antibodies (p = 0.001) correlated to a decreased CD4 cell count. Lack of p24 antibodies was more frequent than was the presence of HIV antigen among persons with decreased CD4 cell count and decreased response to pokeweed mitogen, indicating that absence of p24 antibodies may be an earlier marker of immune dysfunction than the presence of HIV antigen. In persons with p24 antibodies present, a low such titer was associated with a decrease of both immune parameters. Presence of HIV antigen and absence of p24 antibodies thus seems to correlate with the severity of immune dysfunction in healthy HIV infected individuals. PMID- 2970387 TI - Atrial natriuretic peptide in atrial fibrillation before and after electrical cardioversion therapy. AB - In eight patients with atrial fibrillation of less than 3 months duration and without congestive heart failure the plasma concentration of atrial natriuretic peptide was determined one day before, the day after and again 30 days after electrical cardioversion therapy. The pretreatment plasma concentration of the peptide was 99 pg mg-1 (23-480, median and range). The day after cardioversion to sinus rhythm the peptide concentration had normalized to 36 pg ml-1 (18-151). The plasma concentration of atrial natriuretic peptide remained stable in all but one patient for a period of 30 days (46 pg ml-1, 16-695) (P = 0.03). In conclusion, the plasma concentration of atrial natriuretic peptide in patients with atrial fibrillation was significantly reduced after electrical cardioversion to sinus rhythm and remained stable for a period of 30 days. PMID- 2970388 TI - Effects of intracoronary isosorbide dinitrate during acute myocardial ischaemia: a study during angioplasty. AB - The effects of isosorbide dinitrate on acute myocardial ischaemia were studied in 19 patients during coronary angioplasty. The duration of balloon inflation to the onset of ST segment depression was increased by the administration of intracoronary isosorbide dinitrate from (mean +/- SEM) 13 +/- 2 to 20 +/- 2 s (48%) (P less than 0.01) and the duration to 1 mm ST segment depression increased form 23 +/- 2 to 29 +/- 3 s (28%) (P less than 0.01). Systolic blood pressure fell from 118 +/- 3 to 111 +/- 4 mmHg but there was no change in intracoronary distal occlusion pressure. Thus during coronary occlusion isosorbide dinitrate prolongs the time to the onset of myocardial ischaemia. PMID- 2970389 TI - Plasma and right auricle concentrations of atrial natriuretic polypeptide in children with cardiac diseases. AB - To investigate the atrial natriuretic polypeptide (ANP) levels in congenital heart diseases, we measured plasma immunoreactive ANP (ir-ANP) in 29 patients with left-to-right shunt, and right auricle ir-ANP in 25 patients who underwent operation. The levels of ir-ANP in the pulmonary artery correlated with that in the femoral vein (r = 0.95, P less than 0.001), mean pulmonary arterial pressure (r = 0.74, P less than 0.001) and right atrial pressure (r = 0.53, P less than 0.01). The right auricle ir-ANP level was high in patients with heart failure combined with pressure or volume overload of the right atrium, as observed in tricuspid atresia, complete endocardial cushion defect, and total anomalous pulmonary venous drainage. These results suggest that the production of ANP in the atrium increases as a compensatory or secondary response to persistent release of ANP into the circulation in cardiac diseases. PMID- 2970390 TI - Clonal variation in childhood acute lymphoblastic leukaemia at early and late relapse detected by analyses of phenotype and genotype. AB - To increase our knowledge of the clonal relationship of leukaemia relapse, the genotypes and phenotypes of ten children with acute lymphoblastic leukaemia (ALL) were examined at initial diagnosis and relapse. Seven patients were phenotyped as common ALL, two as mixed, and one as T-cell ALL (T-ALL). Comparative analyses of immunoglobulin (Ig) heavy and light chain as well as T-cell receptor beta-chain (T beta) sequences revealed clonal variations, i.e. appearance of a novel or an evoluted leukaemic cell clone in five patients coinciding with the loss of common acute lymphoblastic leukaemic antigen (CALLA) in four cases, irrespective of early or late relapse. Conversion of early B- to T-ALL or lymphoblastic to non lymphoblastic leukaemia was not noted in any of the patients examined. Our results suggest that clonal variation is a frequent event in childhood ALL. PMID- 2970392 TI - Evidence for synergism between the antimyoclonic actions of 5-hydroxytryptophan and clonazepam in rats. AB - The protective effect of the precursor of 5-hydroxytryptamine (5-HT), 5 hydroxytryptophan (5-HTP) against myoclonus induced in rats by picrotoxin and allylglycine was demonstrated. The inhibition by 5-HTP of picrotoxin-induced myoclonic movements was found to correlate well with an increased 5-HT release from the cerebral cortex. p-Chlorophenylalanine (PCPA) pretreatment aggravated the actions of both picrotoxin and allylglycine by shortening their myoclonic latencies. These findings suggest that there is an antimyoclonic effect of 5-HT in the brain. The protective effect of clonazepam against these two myoclonic models was found to be potentiated in 5-HTP-pretreated animals. Only a partial inhibition of its protective effect resulted from PCPA pretreatment. These data suggest that a beneficial synergism is likely to occur between 5-HTP and clonazepam for the inhibition of myoclonus and that a 5-HTergic mechanism does not play a significant role in the antimyoclonic action of clonazepam. PMID- 2970391 TI - Phase I clinical and pharmacokinetic trial of oral menogaril administered on three consecutive days. AB - Eighteen adult patients with solid tumors were treated with oral menogaril, a new anthracycline antibiotic active against human breast cancer after intravenous administration. The drug was given orally on 3 consecutive days every 4 weeks at doses ranging from 50 to 175 mg/m2/day. Reversible and dose-related leukopenia was the dose-limiting toxicity. Thrombocytopenia was less frequent. Hematologic toxicity was maximal usually 2 weeks after treatment and recovery usually occurred within 4 weeks. At doses from 50 to 150 mg/m2/day, non-hematologic side effects of oral menogaril were infrequent and mild and consisted of nausea and vomiting (one patient), alopecia (two patients), mucositis (two patients) and liver function test abnormalities (three patients). The single patient treated at a daily dose of 175 mg/m2/day developed grade IV leucothrombocytopenia, with fever and gastrointestinal bleeding. This was followed by heart failure and the patient died from multisystem organ failure. Peak plasma concentrations of menogaril ranged from 0.043 to 0.409 microM and were linearly correlated with the dose. Similarly, the area under the plasma concentration versus time curve varied from 0.33 to 9.59 microM X h and was linearly correlated with the dose. The mean harmonic half-life was 11.3 +/- 6.4 h. A comparison of the data from the present trial and our previous study with intravenous menogaril indicates a bioavailability of 32 +/- 12%. There was an excellent relationship between the white blood cell decrease (as a percentage of the pretreatment value) and several pharmacokinetic parameters; the best correlation was obtained with the plasma concentration of menogaril at 4 h after treatment. A dose of 150 mg/m2/day for 3 consecutive days is recommended for phase II trials with oral menogaril but the bioavailability of the drug should be monitored carefully and, more specifically, the concept of a pharmacokinetic adjustment of the dose of menogaril should be evaluated prospectively. PMID- 2970393 TI - Echocardiographic abnormalities in acromegalic patients. AB - The aim of our study was the evaluation of the cardiac size and left ventricular function in 26 patients with acromegaly and 20 control subjects using echocardiography. The following parameters were evaluated: systolic and diastolic ventricular dimensions, diastolic posterior wall and interventricular septal thickness, left ventricle ejection fraction and left ventricular mass, amplitude of the posterior left ventricular wall movement, speed of the circular fibres shortening area of the left ventricular muscle and left ventricular contractility index. RESULTS: In 91% of patients with acromegaly echocardiographic abnormalities were found. The most frequent were: concentric thickening of the left ventricular posterior wall in 38%, interventricular septal thickening in 54% and increased left ventricular mass in 84.6% of acromegalic patients. Left ventricular dimensions were increased: diastolic in 65.4% and systolic in 92% of patients. In 46% of cases the ejection fraction was decreased. There was no correlation between extent of the left ventricular hypertrophy and other echocardiographic abnormalities, serum growth hormone concentration, choice of therapy and duration of the disease. PMID- 2970394 TI - Effect of medial preoptic oestradiol implants on hypothalamic beta-endorphin concentration. AB - Twenty-four-day-old immature and cyclic female rats in metoestrus were ovariectomized and bilaterally implanted with oestradiol benzoate (OB) and cholesterol at a ratio of 1:360 or with cholesterol alone in the medial preoptic area (MPOA), or, for control, the hypothalamic dorsomedial nucleus. Estimation of beta-endorphin immunoreactivity in the ventromedial-arcuate-median eminence region at 6 and 3 days after implantation, respectively, revealed a significantly higher concentration in rats implanted with OB in the MPOA as compared to those implanted with cholesterol. OB implants placed in the dorsomedial nuclei were ineffective in this regard. The hypothesis is put forward that inhibition of hypothalamic beta-endorphin release as probably resulting from the implantation of OB in the MPOA may be related to desensitization of the negative oestrogen feedback that is induced by similar preoptic oestrogen implantation. PMID- 2970395 TI - Rapid stimulation of fluid-phase endocytosis and exocytosis by insulin, insulin like growth factor-I, and epidermal growth factor in KB cells. AB - Effects of growth factors on fluid-phase endocytosis and exocytosis in human epidermoid carcinoma KB cells were examined by measuring horseradish peroxidase (HRP) as a marker. Insulin, insulin-like growth factor-I (IGF-I), and epidermal growth factor (EGF) promoted HRP accumulation. They also stimulated the efflux of the preloaded HRP from the cells. From these results it follows that these growth factors stimulate the influx as well as the efflux of HRP, because the accumulation rate is the sum of the influx rate and the efflux rate. The stimulation of both HRP accumulation and HRP efflux was rapidly induced within 2 4 min of the addition of growth factors and persisted for at least 60 min. The concentrations eliciting half-maximal stimulatory effects of insulin, IGF-I, and EGF were about 5 X 10(-7), 1 X 10(-9), and 5 X 10(-10) M, respectively. aIR-3 (anti-type I IGF receptor antibody) completely blocked the stimulation of HRP accumulation by IGF-I but very slightly inhibited the stimulation by insulin. The 528 IgG (anti-EGF receptor antibody) inhibited the stimulation of HRP accumulation by EGF. These results indicated that each of these growth factors stimulates the HRP accumulation mediated by the corresponding (homologous) growth factor receptors. The rapid stimulation of fluid-phase influx and efflux may constitute one of the common early cellular responses to growth factors. PMID- 2970396 TI - Rhodopsin-induced experimental autoimmune uveoretinitis: dose-dependent clinicopathological features. AB - We have studied the clinicopathological features of experimental autoimmune uveoretinitis (EAU) induced in Lewis rats by injection of different doses of rhodopsin and its illuminated form opsin. Rhodopsin consistently appears to be more pathogenic than opsin. Injected in Freund's complete adjuvant and pertussis adjuvant 50 micrograms of rhodopsin induces a frequency of severe EAU similar to 250 micrograms of opsin. Intensity, frequency and location of ocular inflammation are markedly dose dependent. At high dose (100-250 micrograms), rhodopsin induces severe bilateral uveoretinitis in all animals, which starts with acute inflammation of the anterior eye segment at day 10-12 followed by chorioretinitis (predominantly retinitis) which results in complete elimination of the photoreceptor cells. At low dose (20 micrograms), rhodopsin induces mild transient inflammation in 60% of the animals, mainly consisting of mild posterior retinitis which starts at day 20 and leads to a typical multiple focal destruction of the photoreceptor cells. Intermediate doses cause an intermediate type of disease. Omission of pertussis adjuvant lowers the frequency of severe disease at low doses of rhodopsin, delays its onset and changes its features. The last characteristic has been observed in particular at intermediate doses (50-100 micrograms). In these cases, EAU usually starts by cell infiltration of the vitreous, while the anterior segment is only mildly affected. Without pertussis adjuvant the pathogenicity of opsin is low. Even in both adjuvants severe EAU can only be evoked by a high dose of opsin. Although there exists a marked difference in uveitogenicity between rhodopsin and opsin, the immunogenicity is similar and seems not to be correlated with their pathogenicity. PMID- 2970397 TI - [Comparative evaluation of the cardiotropic effect of broncholytic agents]. AB - The cardiotropic effect of broncholytic agents of different groups used for treatment of bronchial asthma was analyzed by using the method of determination of isoenzyme of MB creatine phosphokinase (CPK) and total CPK in the blood of the patients. A total of 125 biochemical assays on determination of CPK in the blood of the patients were made. The cardiotropic effects of three agents (adrenaline, novodrin, Berodual) were studied. Adrenomimetics adrenaline and isodrin (novodrin) were shown to increase significantly CPK activity. A combined agent Berodual insignificantly increased the activity of MB isoenzyme of CPK. PMID- 2970398 TI - [Cardio- and hemodynamic effects of a crown ether derivative and an analysis of its cardiotropic action in an experiment]. AB - The cardio- and hemodynamic effects of the derivative of aza-15-crown-5 was studied in acute experiments on anesthetized (ethaminal sodium, 50 mg/kg) cats. At intravenous administration the compound was found to induce hypotension, to dilate arterial vessels, to exert negative ino- and chronotropic effects, to decrease the cardiac output. The pharmacological analysis suggests that the mechanism of the cardiotropic action of the compound is due to its intervention in calcium ion metabolism. PMID- 2970399 TI - [Effect of nicotinamide on body glucose utilization in animals]. AB - A single administration of nicotinamide (62.5 mg/kg) was found to enhance the relative activity of exogenous insulin, to decrease glucose level in the peripheral blood, to increase the activity of malate dehydrogenase and glucose-6 phosphate dehydrogenase in the liver cytosol in intact mice and to increase the rate of exogenous glucose utilization in rabbits. The character of nicotinamide effect on carbohydrate metabolism in intact animals is suggested to be dose dependent. When administered in low doses, the drug increases tissue sensitivity to endo- and exogenous insulin. PMID- 2970400 TI - The vascular surgeon and transluminal angioplasty. PMID- 2970401 TI - Femoro-popliteal artery thrombolysis with intra-arterial infusion of recombinant tissue-type plasminogen activator--report of a pilot trial. AB - Recombinant tissue-type plasminogen activator (rt-PA) was infused at a rate of 10 mg/h into 50 thrombosed femoral and popliteal arteries. Patency was restored in 43 but a secondary angioplasty led to 2 reocclusions and in 3 patients early rethrombosis occurred. A favourable clinical result was thus obtained in 38 patients (76%). Thirteen bleeding complications occurred in 10 patients, mainly haematomas at puncture sites. One patient required blood transfusion for gastro intestinal bleeding from a previously unknown ulcer. The angiographic recanalisation rate in 16 patients who received a slower infusion of rt-PA (5 or 3 mg/h) was 94% and the clinical success rate in this series was 81%. However, the incidence of bleeding complications was not decreased by the slower infusion rate. The data obtained confirm the feasibility of rt-PA thrombolysis in peripheral arterial thrombosis and warrant a comparative study with streptokinase. PMID- 2970402 TI - [Health education materials in the examination office]. PMID- 2970403 TI - [Pains and the lumbosacral area]. PMID- 2970404 TI - [Self-examination and its role in the early detection of breast cancer]. PMID- 2970405 TI - Carbon dioxide laser laparoscopy by means of a 3.0-mm diameter rigid wave guide. AB - Rigid carbon dioxide wave guides measuring 300 to 500 mm in length and 3.0 mm in diameter were evaluated in rabbits and humans. The wave guide provided an effective laser delivery system for operative laparoscopy. The tested guide was introduced into the abdominal cavity alternatively via the operating channel of a 9-mm operating laparoscope, through the central channel of a smoke evacuation/irrigation cannula, or via a 3.25-mm second puncture trocar. In contrast to previously tested flexible guides, the rigid device transmitted a visible helium-neon aiming beam, power up to 50 watts, and a beam diameter of less than 1 mm. The hollow guide was kept free of smoke by-products by continuous purging with carbon dioxide gas flowing at 800 to 1000 cc per minute. All wave guides could be sterilized by ethylene oxide gas or Cidex (Surgikos, Arlington, TX) soaking. Fourteen women with a variety of reproductive disorders underwent laser laparoscopy. The rigid wave guide was fired at distances ranging from less than 1 mm to 30 mm from the target and performed with maximal impact at distances of 3 to 5 mm. Perhaps the greatest advantage of this system is its ability to focus-defocus while directly coupling to the handpiece of standard lasers and requiring no special alignment procedures. PMID- 2970406 TI - Treatment independent pregnancy with operative laparoscopy for endometriosis in an in vitro fertilization program. AB - Thirty-nine patients with Stage I to IV endometriosis and at least 5 years of primary infertility were enrolled in the Johns Hopkins In Vitro Fertilization Program. At the time of laparoscopic oocyte retrieval, operative endoscopy with lysis of adhesions, fulguration or resection of pelvic endometriosis, or enucleation of ovarian endometriomas was performed. Although the in vitro fertilization-embryo transfer cycle did not result in pregnancy, 12 patients (28%) conceived within 10 months of the operative laparoscopic procedure. Nine of the pregnancies occurred in patients with Stage I to II endometriosis. PMID- 2970407 TI - Comparison between flare up and down regulation effects of luteinizing hormone releasing hormone agonists in an in vitro fertilization program. AB - Luteinizing hormone-releasing hormone (LH-RH) agonists are being increasingly used in ovulation stimulation protocols in IVF programs. The results of two methods of utilization of LH-RH agonists are compared. In the long protocol, gonadotropin stimulation was commenced only after a preliminary period of pituitary desensitization with LH-RH agonist. In the short protocol, exogenous gonadotropins were administered shortly after the start of LH-RH agonist therapy, benefiting from the gonadotropin flare up effect. One hundred eighty-six patients were equally divided between the two treatments. There was no difference in the ovarian response on the day of human chorionic gonadotropin (hCG) or the number of mature oocytes recovered. The cleavage rate of mature oocytes was higher in the short protocol (70% versus 56% P less than 0.01). The ongoing pregnancy rate per treatment cycle was similar in both groups (18% in the long protocol and 16% in the short protocol). Analysis of the luteal phases revealed a trend for higher progesterone values in the long protocol although this was only significant on the 2nd day following oocyte retrieval. As the clinical results were similar, other factors should be taken into account when deciding therapy. These include patient convenience, cost, and side effects. Other schedules of ovulation stimulation with LH-RH agonists are discussed. PMID- 2970408 TI - [Dental prostheses for hemophiliac patients]. PMID- 2970409 TI - [Christ-Siemens-Weech syndrome. Clinical case]. PMID- 2970410 TI - Preliminary report on activity and tolerability of epomediol, administered by intravenous infusion, in patients with chronic hepatopathies. AB - The therapeutic activity and tolerability of epomediol were studied in 28 patients with chronic hepatopathies. Treatment was continued, parenterally, for 10 days (400-600 mg once daily by intravenous infusion). Clinical parameters (headache, right hypochondrial pain, bitter taste in the mouth, asthenia and nausea) and hepatic function (transaminase, alkaline phosphatase and gamma glutamyl transpeptidase) showed significant improvements. Clinical and systemic tolerabilities of epomediol were satisfactory. PMID- 2970411 TI - Beta-endorphin-induced inhibition and stimulation of insulin secretion in normal humans is glucose dependent. AB - This study evaluated the effect of human beta-endorphin on pancreatic hormone levels and their responses to nutrient challenges in normal subjects. Infusion of 0.5 mg/h beta-endorphin caused a significant rise in plasma glucose concentrations preceded by a significant increase in peripheral glucagon levels. No changes occurred in the plasma concentrations of insulin and C-peptide. Acute insulin and C-peptide responses to intravenous pulses of different glucose amounts (0.33 g/kg and 5 g) and arginine (3 g) were significantly reduced by beta endorphin infusion (P less than .01). This effect was associated with a significant reduction of the glucose disappearance rates, suggesting that the inhibition of insulin was of biological relevance. beta-Endorphin also inhibited glucose suppression of glucagon levels and augmented the glucagon response to arginine. To verify whether the modification of prestimulus glucose level could be important in these hormonal responses to beta-endorphin, basal plasma glucose concentrations were raised by a primed (0.5 g/kg) continuous (20 mg kg-1.min-1) glucose infusion. After stabilization of plasma glucose levels (350 +/- 34 mg/dl, t = 120 min), beta-endorphin infusion caused an immediate and marked increase in plasma insulin level (peak response 61 +/- 9 microU/ml, P less than .01), which remained elevated even after the discontinuation of opioid infusion. Moreover, the acute insulin response to a glucose pulse (0.33 g/kg i.v.) given during beta endorphin infusion during hyperglycemia was significantly higher than the response obtained during euglycemia (171 +/- 32 vs. 41 +/- 7 microU/ml, P less than .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970412 TI - [Use of transcranial electrical stimulation for managing the alcohol abstinence syndrome]. PMID- 2970413 TI - [Congenital coronary fistulae. Comments on 9 surgical cases]. AB - The authors relate their experience concerning the surgical correction of congenital coronary fistula. Between May 1971 and June 1986, 9 patients (4 boys and 5 girls) aged from 17 days to 49 years were operated upon at the Division of Cardiac Surgery of Bergamo (Ospedali Riuniti). All the patients, except three who were asymptomatic, showed early cardiac failure or dyspnoea on effort and angina in the elderly. At the physical examination a continuous murmur was heard in 8 cases; the chest x-ray showed significant cardiomegaly and on the electrocardiogram a right/left ventricular hypertrophy pattern was detected in 5 patients. All the patients underwent cardiac catheterization and a coronary angiography. The left-to-right shunt ranged from 60% to 250% of the cardiac output. The anomalous communication affected the right coronary artery in 7 cases and the left in 2, opening into the right atrium in 4 patients, into the right ventricle in 3 and into the pulmonary artery in 2. All patients but one, in whom division and suture were the only necessary procedures, underwent correction by means conventional cardiopulmonary by-pass with moderate hypothermia. In 3 cases closure through the coronary artery was preferred, in 1 through the right ventricle and in 2 transpulmonary. There was only 1 late death which occurred in a 3 year-old patient due to renal failure. After a mean follow-up of 6 years, 7 patients are to be asymptomatic while 1 patient had to be reoperated for a significant residual shunt. PMID- 2970414 TI - [Dermatopathies in the domestic worker: the Lombard experience]. AB - In order to identify the etiopathogenesis of cutaneous diseases in housewives, a questionnaire was made, and the gathered data are here described. 1,824 people have answered this questionnaire: 1,007 have reported lesions with a suspicious immunological pathogenesis. The commercial products which were reported as a cause of the dermatitis have been 834. In a group of 142 housewives (86 with allergic contact dermatitis and 56 with irritative contact dermatitis), the allergic sensitizations to the chemicals in the detergents were tested. 21% of the cases had positive cutaneous reactions for nickel salts and 8.4% for chromium salts. The analysis in atomic absorption method of these metals in 34 commercial products has shown their presence in almost all the samples. Hypothesis are formulated to find the source of these polluting substances. The aims and the methods of a investigation carried out on 1.800 housewives, who are going to be controlled in order to estimate the prevalence, the clinical kinds and the etiopathogenesis of dermatitis related to house work, are described. The study will be carried out at the Occupational Health Institute "Clinica del Lavoro L. Devoto" of Milan and at the Dermatological Departments of the Universities of Bari and Florence. PMID- 2970415 TI - [Dermatopathies in those engaged in domestic work and related risks]. AB - The AA., considering the skin diseases in the subjects employed in the domestic work and similar activity, in a more ample table of general medicine, produced a contribution to the study of etiopathogenetic and nosographic problems. They have estimated the role of the individual predisposition and the etiopathogenetic, chemical, physical, biotic, complex factors. They pointed out the various nosographic, skin expressions in range of contact irritant dermatitis, of contact allergic eczema, skin abrasion, of dermatitis by physical agents, of urticaria, of dermal alterations by infectious and parasitic agents and of various dermal, morbid expressions by chemical agents. They reported the direct experience of personal casuistry and called attention on multiple aspects of preventive medicine. PMID- 2970416 TI - Listeria peritonitis diagnosed by laparoscopy. PMID- 2970417 TI - Percutaneous endoscopic gastrostomy feeding tube migration and impaction in the abdominal wall. PMID- 2970418 TI - [Improvement in the blood service in the Georgian SSR]. PMID- 2970420 TI - Purification and characterization of the DNA-binding protein Ner of bacteriophage Mu. AB - The construction is described of a plasmid (pL-ner) which directs the high-level production of the bacteriophage Mu Ner protein in Escherichia coli. The protein, recovered in the soluble cellular fraction, was susceptible to in vivo proteolytic processing, in many host strains, but not in E. coli B, a natural lon prototroph. A simple purification method is described which takes advantage of the basic nature of the protein. The purified protein was shown to be physically and chemically homogeneous and to have an amino acid sequence identical to that predicted for the authentic protein. The protein was also shown to have in vitro biological activity, as measured by specific binding to a DNA fragment containing the consensus Ner-binding sequence, and in vivo biological activity as the protein produced by the pL-ner plasmid allowed lysogenic-like maintenance of a Mu prophage c mutant unable to synthesise a functional Mu repressor. PMID- 2970419 TI - Adaptation at specific loci. V. Metabolically adjacent enzyme loci may have very distinct experiences of selective pressures. AB - The polymorphic phosphoglucomutase (PGM) and glucose-6-phosphate dehydrogenase (G6PD) loci have been studied in parallel to experimental work on the phosphoglucose isomerase (PGI) polymorphism in Colias butterflies. PGI, PGM and G6PD are also autosomal in Colias. PGM and G6PD are loosely linked (and represent the first identified autosomal linkage group in Colias); they assort independently from PGI. Recombination occurs in both sexes. Neither PGM nor G6PD shows large, consistent differences in flight capacity through the day among its genotypes, as PGI does. PGM shows some change of allele frequencies, and match to Hardy-Weinberg expectation, with air temperature in middle and latter parts of the season, but not early in the season. G6PD may show some heterozygote excess over Hardy-Weinberg expectation early in the day, but more testing is needed. No evidence for differential survivorship was seen at PGM or G6PD, in contrast to PGI. At the PGM and G6PD loci, male heterozygotes are advantaged in mating with females, but without the evidence of female choice which occurs for PGI. These effects are not correlated among the three loci. There is no assortative mating at G6PD (nor at PGI). There is minor positive assortative mating of PGM heterozygotes, but it is too weak to account for the PGM-genotype-specific male mating advantage. No trends of multilocus genotype frequencies involving PGI are seen. Certain PGM-G6PD two-locus genotypes are over-represented, and others under represented, in wild adult samples, particularly among males and uniformly among successfully mating males. Our results emphasize that enzyme loci sharing a substrate need not have common experience of the existence or strength of natural selection, and suggest initial food-resource processing and allocation as a possible context for fitness-related effects of the PGM and G6PD polymorphisms. PMID- 2970421 TI - Expression of cloned genes by in vivo insertion of tac promoter using a mini-Mu bacteriophage. AB - The strong trp-lac(tac) promoter has been incorporated into the mini-Mu bacteriophage genome to form a mini-Mu-tac (mMu-tac) expression transposon. This mMu-tac element can transpose efficiently in Escherichia coli cells when derepressed and occasionally insert into a recombinant plasmid. When mMu-tac integration occurs upstream of a cloned gene in the orientation of its transcription, the tac promoter can direct the expression of the gene insert. mMu tac contains translation stop codons downstream of the tac promoter. Thus, mMu tac should be useful to express only those genes containing their own translational information. We report here the successful expression in E. coli of several prokaryotic genes using the transposon expression system. PMID- 2970422 TI - Two human genes encoding tRNA(GCCGly). AB - Two human DNA fragments of 16.7 and 15.5 kb have been selected from a human lambda Charon-4A library by hybridization to an unfractionated tRNA probe. Restriction mapping and Southern and Northern hybridization analyses revealed the presence of a single tRNA-hybridizing region in each of the human DNA fragments. Nucleotide sequence analysis has identified two identical members of the tRNA(GCCGly) gene family. These tRNA(GCCGly) genes encode all of the conserved and semiconserved nucleotides of the tDNA split promoter sequences. Neither gene contains introns or encodes the CCA sequence present on the 3' terminus of mature tRNA. One of these identical tRNA(GCCGly) genes was found to be expressed at a substantially greater efficiency than the other in a HeLa cell lysate in vitro transcription system. No similarity was detected in the nucleotide sequences flanking these genes other than the characteristic, 3' oligo[dT] transcription termination signals and a TCTTT sequence located 7 to 10 bp upstream. These data are consistent with the hypothesis that as yet unidentified tDNA flanking sequences may have an important role in modulating human tRNA gene expression. PMID- 2970423 TI - A cytogenetic study of mentally retarded school children in Taiwan with special reference to the fragile X chromosome. AB - A cytogenetic study was made on 341 mentally retarded children in the Provincial Nantou Rehabilitation Center for the Mentally Retarded and the St. Raphael Opportunity Center in Tainan. Of the 89 mentally retarded children with chromosomal abnormalities, 63 had Down syndrome, 13 had the fragile X [fra(X)] syndrome, and the remaining had other aneuploid constitutions. Family studies were possible for 2 of the 13 fra(X) probands. The results of this study illustrate the contribution of chromosomal abnormalities to the pathogenesis of mental retardation in children. PMID- 2970424 TI - Expression of fragile sites in childhood acute lymphoblastic leukemia patients and normal controls. AB - A high concordance has been reported between fragile sites and breakpoints involved in chromosomal rearrangements in cancer. A prospective study on the role of fragile sites in the etiology of childhood acute lymphocytic leukemia (ALL), with appropriate comparisons to results obtained from normal controls, analyzed fluorodeoxyuridine-, aphidicolin-, and caffeine-induced fragile sites in the peripheral blood of seven ALL patients (three with cytogenetically normal karyotype and four with pseudodiploid karyotype) and eight normal controls. While extensive variations in the number and distribution of fragile sites was observed within each group, there was no significant difference in the mean total fragile sites and mean fragile sites per cell between the two groups (P greater than 0.05) in all three treatments. Similarly, within the ALL patients, the two karyotypic groups did not exhibit any significant difference in fragility (P greater than 0.05). PMID- 2970425 TI - The role of fluorinated pyrimidine analogues in the induction of the in vitro expression of the fragile X chromosome. AB - The modes of action of 5-fluoro-2'-deoxyuridine (FdUrd) and 5-fluoro-2' deoxycytidine (FdCyd) were studied in PHA-stimulated lymphocytes from normal volunteer donors and a fragile X patient. In both cell types, FdUrd and FdCyd inhibited cell proliferation at concentrations of 3 x 10(-8) M. Thymidylate synthetase was identified as the decisive target for the action of both FdUrd and FdCyd, as judged from the following observations: First, addition of thymidine to the culture medium was able to counteract both FdUrd and FdCyd toxicities, whereas addition of dCyd had no observable effect. Second, inhibition of the in situ thymidylate synthetase activity measured as an increase in the level of [3H] dThd incorporation coincided with the inhibition of cell proliferation. Third, the inhibition of the thymidylate synthetase-dependent incorporation of [3H]-dUrd into newly synthesized DNA coincided with the inhibition of cell proliferation. The effects of FdUrd and FdCyd on the in vitro expression of fragile site Xq27 of fragile X chromosomes was shown to be based on the depletion of the intracellular pool of thymidine-5'-monophosphate (dTMP), as judged from the following observations: First, both the FdUrd- and FdCyd-dependent induction of site Xq27 coincided with the antiproliferative effects of the respective fluoropyrimidines. Second, addition of thymidine (dThd) to the culture medium both prevented the expression of site Xq27 and neutralized the cytotoxicity of FdUrd and of FdCyd. On the basis of these findings, we provide further evidence for the concept that the fragile X site is located in an AT-rich region. PMID- 2970426 TI - Breakage of the T cell receptor alpha chain locus in non malignant clones from patients with ataxia telangiectasia. AB - Patients with ataxia telangiectasia (A-T) develop specific chromosome translocations, which may confer a proliferative advantage, resulting in the appearance of large clones in the peripheral blood lymphocytes. These lymphocytes are not malignant. Using in situ hybridisation techniques we have investigated a consistent 14q11 translocation breakpoint observed in a t(X;14)(q28;q11) translocation clone from each of two different patients and a t(14;14)(q11;q32) clone from a third patient. In all cases the chromosome translocation involved breakage within the alpha chain locus of the T cell receptor (TCR alpha), between the variable and constant regions, at 14q11. Chromosome rearrangement involving breakage within TCR alpha can therefore precede the development of malignancy. Further chromosomal rearrangement may be required in these patients, for progression to the leukaemic state. PMID- 2970427 TI - Biotypes, serotypes, and susceptibility to antibiotics of 60 Haemophilus influenzae strains from genitourinary tracts. AB - Sixty strains of Haemophilus influenzae were isolated from the genitourinary tracts of adults: 19 from cervicovaginal secretions, one from a woman with bartholinitis, 37 from urethral exudates, and three from urine. Non-capsulated strains were recovered predominantly, and biotype III accounted for 28 isolates and biotype IV for 25. Many of the H influenzae strains were found to be resistant to one or more of the antibiotics commonly used against sexually transmitted diseases. Resistance to tetracycline was prevalent and was found in 17 of the strains. Ten strains were ampicillin resistant and beta lactamase producing. Kanamycin resistance was less common (two strains). Trospectomycin (U 63366), a new spectinomycin analogue, was eight to 16 times more active than spectinomycin. All the quinolones tested were very active against all strains and may provide an effective alternative for the treatment of resistant H influenzae in genitourinary infections. PMID- 2970428 TI - Binding and catabolism of aggregated immunoglobulins bearing C3b or iC3b by U937 cells. AB - Mononuclear cells play an important role in the elimination of immune complexes (IC). In the presence of complement (C) the binding and degradation of IC by mononuclear cells is enhanced at least two-fold. The enhancement of binding is caused by a synergistic interaction of the IC with cellular Fc and complement receptors (R). In the present study we have investigated the contribution of the complement receptors CR1 and CR3 of human monocyte cell line U937 on the complement-mediated binding and degradation of immune complexes and soluble aggregates of IgG (AIgG) bearing C3b or iC3b. It was found that deposition of C3b on AIgG enhanced the binding of AIgG to U937 cells at least two-fold. The C3b mediated enhancement of binding was abolished by anti-CR1. iC3b-bound to AIgG also enhanced the binding of AIgG to the cells. This binding was only partially reduced by anti-CR3 antibodies, but the combination of anti-CR1 and anti-CR3 fully abolished the iC3b-mediated enhancement of binding. These results suggest that both CR1 and CR3 contribute to the complement-mediated binding and degradation of soluble IC by mononuclear phagocytes. PMID- 2970429 TI - Suppressor T-cell population induced by Junin virus in adult mice. AB - Intracerebral (i.c.) Junin virus (JV) infection of adult BALB/c mice is characterized by the absence of morbidity and a low mortality (barely 8-10%). In contrast, the suckling mouse model exhibits almost 100% mortality following central nervous system (CNS) alterations consistent with a delayed-type hypersensitivity (DTH)-like immune response. Besides, JV infection of adult (resistant) mice leads to immunosuppression of DTH to unrelated antigens. Here we present evidence demonstrating that such suppression is mediated by JV-induced cells present in spleen from 24 hr to 24 days post-infection, bearing the Thy-1+, Ly-1+2- phenotype and reactive to an unrelated antigen such as sheep red blood cells (SRBC). No evidence of suppressor factors was found. A relatively low number of total splenic cells (5 x 10(6) cells/mouse) was enough to transfer suppression. Therefore, this cell population may be involved in adult mouse survival to JV infection. PMID- 2970431 TI - Purified human NK cells do not function as accessory cells in T-cell proliferative responses. AB - In the present study we investigated the accessory function of natural killer (NK) cells in PHA, anti-CD3 (OKT3) and solid-phase-bound anti-CD3-induced T-cell proliferative responses, as well as in allogeneic mixed lymphocyte reaction (MLR). Although NK cell-enriched Percoll fraction cells showed accessory activity in all of the studied T-cell responses, the accessory function in anti-CD3 and allogenic responses were always found to be strictly dependent on the proportions of contaminating monocytes (Mo) and dendritic cells (DC). Purified CD16+ NK cells were totally incapable of inducing T-cell proliferation in anti-CD3, solid-phase bound anti-CD3 and in allogeneic MLR responses, whereas these NK cells could support PHA-induced T-cell proliferation, although to a very low extent. Growing evidence suggests an inhibitory role of NK cells in various immune responses, but in the studied T-cell responses NK cells did not suppress either Mo or DC accessory ability. PMID- 2970430 TI - The effects of pertussis toxin on human T lymphocytes. AB - Purified pertussis toxin (PPT) is a potent mitogen for human T lymphocytes and is shown to cause rapid calcium mobilization in resting T cells, a T-cell line and CD3- lymphocytes with natural killer (NK) activity. In resting T cells the PPT activation is associated with cytoplasmic alkalinization. A similar rise in intracellular free calcium ([Ca2+]i) and cytoplasmic alkalinization is observed with activation through the antigen receptor complex. The effect of PPT is unlikely to be mediated through this pathway, however, as it can mobilize calcium in lymphocytes that do not express the CD3-Ti complex. In contrast to several other cell types, re-incubation of resting human T cells with PPT, up to a dose of 100 ng/ml for 2 hr does not block subsequent agonist-induced calcium mobilization dependent on G protein-mediated phospholipase C activation. Mitogenic doses of PPT cause a modest reduction in subsequent agonist responses, but this is likely to be due to a post-activation refractory state rather than G protein inactivation. PMID- 2970433 TI - Significance of renal vasodilation after administration of atrial natriuretic factor in the conscious dog. AB - The contribution of alterations in renal hemodynamics to the diuretic and natriuretic actions of atrial natriuretic factor (ANF) was studied in chronically instrumented conscious dogs. Injection of ANF-(99-119), 10 micrograms/kg, had no effect on mean arterial pressure, heart rate, renal blood flow, or calculated renal vascular resistance; However, it increased urine flow rate (86 +/- 20%) and sodium (118 +/- 24%) and potassium (35 +/- 22%) excretion (p less than 0.05). In contrast, ANF-(99-122), 10 micrograms/kg, significantly increased renal blood flow (26 +/- 4.5%), reduced renal vascular resistance (24 +/- 2.9%) and arterial pressure (5.5 +/- 1.9%), and markedly increased urine flow rate (198 +/- 34%) and sodium (206 +/- 32%) and potassium (75 +/- 27%) excretion (p less than 0.05), being almost twice as effective in the first 10 minutes as was ANF-(99-119) infusion. During a brief infusion, ANF-(99-122) (10 micrograms/kg/min for 4 minutes) increased renal blood flow (24 +/- 2.7%), heart rate (18 +/- 5.7%), urine flow rate (199 +/- 25%), and sodium (290 +/- 81%) and potassium (104 +/- 17%) excretion. Injection of radioactive microspheres (15 or 9 micron) to measure intrarenal distribution of blood flow during the steady state increase in renal blood flow indicated that ANF-(99-122) infusion preferentially increased outer cortical blood flow. Blood flow in the four zones of the kidney cortex (Zone 1, outer, and Zone 4, inner) increased 96 +/- 25% (Zone 1), 199 +/- 87% (Zone 2), 139 +/- 47% (Zone 3), p less than 0.05, and 25 +/- 28% (Zone 4, p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970432 TI - Expression of Fc gamma receptor-mediated phagocytosis by airway intra-luminal macrophages. AB - Macrophages in the conducting airways of the lower respiratory tract constitute an anatomically defineable subpopulation of pulmonary macrophages. Little information regarding the functional characteristics of the airway intra-luminal macrophages (AI-LM) is currently available. In this study, the AI-LM resident in the trachea and mainstem bronchi of the rat were harvested by an airway lavage technique and the ability of the AI-LM to phagocytize by Fc gamma-receptors was evaluated relative to the phagocytic activities of alveolar macrophages (AM) obtained by bronchoalveolar lavage. More than 60% of the AI-LM phagocytized sheep erythrocytes opsonized with IgG (SRBC-IgG) and the distributions of the engulfed SRBC-IgG in the phagocytic AI-LM were virtually identical to those in AM. The AI LM may represent AM that have been translocated from the alveolar space compartment to the conducting airways; it remains possible, however, that at least some of the AI-LM may have an airway origin. Regardless, the results of our phagocytic studies suggest AI-LM may functionally provide a protective phagocytic role in the conducting airways. PMID- 2970434 TI - Dietary sodium change in primary aldosteronism. Atrial natriuretic factor, hormonal, and vascular responses. AB - Atrial natriuretic factor (ANF) may be physiopathologically involved in several clinical conditions including human hypertension. However, few data are available regarding this putative hormone and its relationship to aldosterone, blood pressure, and vascular responsiveness to alpha-adrenergic receptor stimulation in primary aldosteronism, a volume-expanded, low-renin model of human hypertension. For this reason, the behavior of supine and upright plasma ANF as related to aldosterone, blood pressure, and forearm alpha-adrenergic sensitivity (plethysmographic technique) to intra-arterial norepinephrine infusion was studied in eight patients with primary aldosteronism (five with adenomas, three with hyperplasia) before and at the end of two sequential 1-week low (20 mmol/day) and high sodium (200 mmol/day) diet periods. Basal, predict ANF concentrations decreased and increased after low and high sodium intakes, respectively. Furthermore, highly significant postural ANF decrements after 1 hour of standing occurred with each diet, although they were lower after the low than after the high sodium diet. Plasma aldosterone, either supine or upright, was insensitive to dietary sodium manipulations, suggesting the absence of ANF mediated control of aldosterone secretion in our patients. In spite of about twofold higher ANF concentrations during the high than during the low sodium diet, forearm vascular sensitivity to intra-arterial norepinephrine infusion did not change during the study. Furthermore, systemic arterial blood pressure rose to a highly significant extent after dietary sodium content was increased, thus casting doubt on a role for ANF as an endogenous long-term modulator of systemic blood pressure and peripheral alpha-adrenergic sensitivity in patients with primary aldosteronism. PMID- 2970435 TI - Adherence of Streptococcus sanguis to conformationally specific determinants in fibronectin. AB - The adherence of Streptococcus sanguis to specific receptors exposed or deposited at the site of endothelial damage may play an important role in the development of infective endocarditis. Adherence of the Challis strain of S. sanguis to gelatin (or collagen) and gelatin-binding components of plasma was examined with an enzyme-linked immunosorbent assay. S. sanguis adhered poorly to immobilized gelatin and to molecular or fibrillar collagen. However, in the presence of fresh human plasma, the adherence of S. sanguis to all three substrates increased as much as eightfold. Removal of gelatin-binding proteins eliminates the ability of plasma to enhance adherence of S. sanguis to the substrates. Addition of purified human plasma fibronectin (Fn) to the absorbed plasma restored the adherence promoting ability in a dose-dependent manner. A similar dose-dependent increase in S. sanguis adherence was observed when increasing concentrations of Fn alone were added to the gelatin-coated assay wells. S. sanguis adherence to immobilized fibronectin could not be inhibited by preincubating either the bacteria or the gelatin-coated assay wells with Fn or by including excess soluble Fn in the assay mixture. Studies with peptides purified from trypsin digests of Fn indicated that the 160- to 180-kilodalton (kDa) fragments which retain both the gelatin-binding and the cell-binding regions of the intact molecule support adherence of S. sanguis to gelatin. The 160- to 180-kDa fragments inhibited the interaction of S. sanguis with immobilized Fn. In contrast, intact Fn and the 31-kDa amino-terminal fragment were unable to inhibit the adherence when used in equivalent or greater molar amounts. These in vitro results suggest that in the presence of whole plasma, S. sanguis binds to immobilized gelatin or collagen via Fn bound to the immobilized substrates. Our finding that adherence of S. sanguis to immobilized Fn can occur in the presence of large concentrations of Fn, whether in plasma or purified, indicates that a S. sanguis-binding domain is cryptic in the Fn molecule while in solution and is exposed by a conformational change when the Fn becomes bound to gelatin-coated plastic. The ability of peptide fragments of Fn to inhibit S. sanguis adherence is consistent with this hypothesis. PMID- 2970437 TI - The suppressive effect of histamine on anti-DNP antibody production in mice. AB - To clarify the effect of histamine on antibody production, histamine was injected intravenously into DNP-KLH immunized C3H/HeN female mice. The number of anti-DNP plaque-forming splenic cells decreased in immunized and histamine-injected mice. Decreased PFC responses similarly occurred in recipient mice into which Lyt-2.1 T cells obtained from histamine-injected mice were transferred before immunization. The effect of histamine is not due to inhibition on plaque formation. The results indicated that histamine induced the augmentation of suppressor T cell (Ts) function and caused the decrease of antibody production in histamine-injected mice. The maximum activity of Ts in recipient mice was obtained from mice injected with histamine 3 days before the cell transfer. The suppressive activity of histamine was not antigen specific because Ts was induced in histamine injected, nonimmunized mice. The treatment of histamine type 2 (H2) agonist showed decreased PFC responses in the mouse as well as histamine did. When mice were treated with an H2 antagonist followed by histamine, antibody production was not reduced. The above results indicated that histamine seemed to regulate antibody production through H2 receptors on lymphocyte membranes. PMID- 2970436 TI - Differential effects of human alpha and gamma interferon on mixed lymphocyte culture and on T-cell-mediated cytotoxicity. Inhibition of proliferation but not of IL-2 production by alpha interferons. AB - We compared the effects of natural and recombinant (r) alpha (IFN-alpha) and gamma (IFN-gamma) interferons on the proliferative responses of human peripheral blood mononuclear cells to mitogens and allogeneic cells in mixed lymphocyte culture (MLC) and on the generation of specific T-cell-mediated cytotoxicity. In 14 of 19 donors, natural IFN-gamma and rIFN-gamma had no significant effect on the proliferative responses to mitogens or allogeneic cells in MLC, even at very high IFN-gamma concentrations (10,000 U/ml). In the remaining 5 donors, a statistically significant (p less than 0.001) enhancement by 49 +/- 8% of the proliferative responses was observed. In contrast, natural IFN-alpha and rIFN alpha 2 significantly inhibited (p less than 0.001) proliferative responses to mitogens and to allogeneic cells, even at concentrations as low as 10 U/ml, in agreement with previous reports. Although natural and recombinant IFN-alpha significantly inhibited these proliferative responses, they did not affect interleukin-2 (IL-2) production in these cultures, suggesting that they inhibit proliferation by a mechanism that does not involve inhibition of IL-2 production. rIFN-gamma did not affect the generation of specific cytotoxicity in MLC, although it was significantly enhanced by natural IFN-alpha and rIFN-alpha 2. Additionally, we compared the ability of human rIFN-alpha subtypes to inhibit proliferative responses to allogeneic cells in MLC. rIFN-alpha 2, rIFN-alpha 4, and rIFN alpha 7 displayed the most potent inhibitory activity of allogeneic responses and were active at concentrations as low as 0.3-0.6 ng/ml.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970438 TI - 1,25-Dihydroxyvitamin D-induced suppressor cells in uremic versus normal lymphocytes. AB - 1,25-Dihydroxyvitamin D3 (DHD) has been shown to suppress mitogen-induced blast transformation. This inhibition is abolished by prior elimination of adherent cells. Chronic renal failure is an immunodeficiency state on the one hand and is associated with abnormalities in vitamin D metabolism on the other. The effect of DHD on the induction of suppressor cells in uremic vs. normal peripheral blood mononuclear cells was investigated. Study groups included 16 chronically uremic patients and 16 age- and sex-matched controls. DHD induced suppressor cell activity in normal lymphocytes. However, no suppressor cell activity was observed in lymphocytes from the uremic patients preincubated with DHD. The origin of the responder cells (normal or uremic) did not affect the outcome. The results would suggest that monocyte-adherent suppressor cells from uremic subjects are either incapable of binding DHD or fail to mount a normal post-receptor intracellular chain of events culminating in suppressor activity. PMID- 2970439 TI - Inhibition of spermatogenesis in men using various combinations of oral progestagens and percutaneous or oral androgens. AB - Eight men (experiment 1) requesting male contraception received a daily oral dose of 20 mg medroxyprogesterone acetate (MPA) combined with 125 mg percutaneous dihydrotestosterone (DHT). Three months later the mean sperm count was only diminished slightly; the replacement of DHT for four men by percutaneous testosterone at the same concentration led to a dramatic fall in sperm count. For 6-18 months all men were treated with MPA plus percutaneous testosterone (250 mg daily). The latter dose restored physiological levels of plasma testosterone. Follicle-stimulating hormone levels were inhibited more severely than in the DHT treated group, whereas LH levels were variable. Azoospermia was achieved and maintained in six cases; two men were oligozoospermic and in one case a moderate secondary rise in the sperm count was observed. Twelve volunteers (experiment 2) received a daily oral dose of either 5 or 10 mg norethisterone acetate plus percutaneous testosterone (250 mg daily). All of them achieved azoospermia within 2 months, but two subjects later exhibited a partial restoration in sperm count. Follicle-stimulating hormone and LH levels were inhibited more severely than in the first experiment. The sperm count and gonadotrophin levels returned to initial values within 6 months after cessation of the treatment in both experiments. No side-effects were noted concerning blood parameters, libido or body weight. However, several female partners had elevated levels of plasma testosterone. In experiment 3 (13 volunteers), percutaneous testosterone was replaced by oral testosterone undecanoate (160 mg daily). Only seven men were azoospermic and most of them had lowered levels of plasma testosterone. Thus, the combination of percutaneous testosterone and oral progestagens appears to be the most convenient for male hormonal contraception. PMID- 2970440 TI - Three-dimensional visualization of velocity profiles in the ascending aorta in humans. A comparative study among normal aortic valves, St. Jude Medical and Starr-Edwards Silastic Ball valves. AB - Blood velocities were recorded intraoperatively in the ascending aorta approximately 6 cm downstream of the aortic annulus in eight patients with normal aortic valves (prior to coronary bypass surgery) and in seven patients after implantation of a St. Jude Medical valve (4) or Starr-Edwards Silastic Ball valve (3) in the aortic position by means of constant temperature hot-film anemometry. The velocities were measured during 5-15 heart cycles at 41 predetermined measurement points almost evenly distributed in the cross sectional area. Velocity profiles were visualized three-dimensionally at 100 time intervals during one mean heart cycle. The standard deviation of velocities was correspondingly three-dimensionally visualized. For the patients with normal aortic valves the velocity profiles were generally flat but skew, with the highest velocity closer to the left vessel wall at the start of systole. During systole the skewness turned counter-clockwise with the highest velocities closer to the posterior, right and finally anterior vessel wall at the end of systole. In patients with an artificial heart valve in the aortic position the design of the valve was reflected on the contour of the profiles. The findings correspond well with model studies. PMID- 2970441 TI - Analysis of velocity in the ascending aorta in humans. A comparative study among normal aortic valves, St. Jude Medical and Starr-Edwards Silastic Ball valves. AB - To analyze velocity spectral energy distribution in humans, blood velocities were recorded by means of hot-film anemometry at 41 predetermined measurement points in the cross-sectional area of the ascending aorta approximately 6 cm downstream of the aortic valves. Measurements were made in 8 patients with normal aortic valves, in 4 after insertion of a St. Jude Medical (SJM) aortic valve and in 3 after insertion of a Starr-Edwards Silastic Ball (SSB) aortic valve. Data analysis based on Fast Fourier Transform demonstrated that turbulence energy was lower in patients with normal aortic valves than in patients after insertion of an artificial valve in the aortic position and probably more pronounced after SSB valves than after SJM valves. The spatial distribution of the turbulence energy above 100 Hz was more irregular than corresponding laminar velocities previously presented. The VER100 (Velocity Energy Ratio at 100 Hz, i.e. the velocity energy above 100 Hz divided by the total velocity energy) proved useful for evaluating differences in flow disturbances downstream of different aortic valves. The mean VER100 in the three categories of patients were respectively 0.3, 1.4, and 2.1%. PMID- 2970442 TI - Effects of chronic administration of the fixed combination slow-release oxprenolol-chlorthalidone on left ventricular hypertrophy in hypertensive patients. Echocardiographic study. AB - To evaluate the effects of the chronic administration of the fixed combination slow-release, oxprenolol 160 mg and chlorthalidone 20 mg on left ventricular hypertrophy, ten hypertensive out-patients, with left ventricular hypertrophy documented by left ventricular mass index (LVMI) greater than 130 g/m2, were studied. After a two-week placebo period, patients were given the study medication, once daily for six months. Blood pressure and heart rate were measured, 24 h after-dosing, at the end of placebo and thereafter every month. A m-mode echocardiographic examination was performed at the end of the placebo period, after 1 month of active treatment and at the end of the study. In comparison with placebo, the study medication induced a significant reduction (p less than 0.01) of systolic and diastolic blood pressure, 24 h after dosing, after 1 month of treatment (from 181.0 +/- 18.5/108.5 +/- 12.0 to 146.5 +/- 10.8/94.0 +/- 7.7 mmHg), and this reduction was maintained until the end of the study (142.0 +/- 14.0/90.0 +/- 8.2 mmHg). At the 6th month and in comparison with placebo, a significant (p less than 0.01) reduction of left ventricular mass (LVM) and of LVMI was observed (LVM: from 295.9 +/- 113.8 to 221.5 +/- 66.1 g; LVMI: from 158.1 +/- 39.0 to 126.2 +/- 35.8 g/m2. In conclusion, our results confirm the good antihypertensive efficacy of the fixed combination slow-release oxprenolol and chlorthalidone and show that the study medication is able to induce a regression of left ventricular hypertrophy, in hypertensive patients. PMID- 2970443 TI - The effect of pirenzepine on the functions of human platelets. AB - Despite the very high chemical similarity of pirenzepine (P) to the selected tricyclic antidepressive drugs (TCAD) (imipramine, desimipramine, nortriptyline and amitriptyline), P has a distinctly different effect on platelets, in particular, on serotonin uptake and platelet aggregation. Using two groups of nine healthy volunteers, mean age 27.7 and 27.5 years, respectively, assessment of the two indicators in vitro helped us to eliminate the possibility of a drug induced defect of the two thrombocyte functions, i.e. their aggregation and serotonin uptake. There is therefore, no reason to fear the danger of decreased hemocoagulation in ulcer patients treated with P for gastrointestinal hemorrhage. The results obtained in vivo using two tests which showed the level of platelet activation associated with release of a specific platelet protein (beta thromboglobulin in platelet-poor plasma) and malondialdehyde in thrombocyte-rich plasma ruled out any changes, specifically their increase, and have disproved any kind of stimulating activity of P on thrombocyte reactivity. PMID- 2970444 TI - Suitability of cyproterone acetate in the treatment of osteoporosis associated with athletic amenorrhea. AB - The effectiveness of the antiandrogenic agent cyproterone acetate (CA) in its contraceptive form (2 mg CA + 50 micrograms ethinyl estradiol) in the treatment of osteoporosis associated with athletic amenorrhea was studied in seven high performance athletes. Four women with similar characteristics served as controls. Their mean age was 21.9 years +/- 3.9. Training was started at a mean age of 14.0 years +/- 2.0. The mean training intensity expressed as kilometers run per week was 35 +/- 15. Mineral density was primarily affected by the hypoestrogenic status of these athletes (= 22 pg/ml +/- 8.8 in the midluteal phase). All participants showed low serum progesterone (= 2.85 ng/ml +/- 2.10) and LH profiles (= 5.6 mlU/ml +/- 0.8) during the midluteal phase. Cyproterone acetate was administered for 8 months to treat the increased bone loss in seven women athletes. Vertebral density appeared to be increased with 9.5% +/- 2.45% (mean +/ SD) while cortical base mineral content measured at the radius was not significantly changed. Our results demonstrate that cyproterone acetate administered in combination with estrogens provides a suitable therapeutic agent in the management of osteoporosis due to a hypoestrogenic status. This treatment could substitute other contraceptive agents. Moreover, women with the most severe estrogen deficiency showed a more pronounced reaction to this therapy. PMID- 2970445 TI - Radiological findings in the lumbar spine of 39 young cross-country skiers with low back pain. AB - The lumbar spines of 39 active young cross-country skiers suffering from long lasting or frequently remitting low back pain (LBP) were examined by a new radiological method. Plain radiographs in lateral and anteroposterior views were taken. The spinal curves, anomalies, and possible degenerative changes were investigated. The radiological method chosen proved to be suitable for imaging the lumbar spine. There were various radiologically detectable abnormalities in the spine. The most common findings were functional scoliosis (in 49%) and different spinal anomalies (in 23%). The mean lumbar lordosis was 69 degrees (70 degrees in males and 64 degrees in females). The effect of abnormal lumbar posture on LBP should be further studied. None of the numerous radiological findings had an effect on therapy in these skiers. PMID- 2970446 TI - The probability of causation approach. AB - Probability of causation (PC) is sometimes viewed as a great improvement by those persons who are not happy with the present rulings of courts in radiation cases. The author does not share that hope and expects that PC will not play a significant role in these issues for at least the next decade. If it is ever adopted in a legislative compensation scheme, it will be used in a way that is unlikely to please most scientists. Consequently, PC is a false hope for radiation scientists, and its best contribution may well lie in some of the spin off effects, such as an influence on medical practice. PMID- 2970447 TI - Effect of neonatal denervation on the distribution of fiber types in a mouse fast twitch skeletal muscle. AB - This study was designed to assess the changes in fiber-type distribution of the extensor digitorum longus (EDL) muscle of the mouse during the first 21 days of age following neonatal sciatic neurectomy. Denervated and normal muscles were compared at 7, 14, and 21 days of age and the normal EDL was also studied at 1 day of age. Frozen sections of the EDL were treated histochemically to detect NADH-tetrazolium reductase and myosin ATPase reactions. Quantitative assessment included measurements of cross-sectional areas and fiber counting. Denervation resulted in muscle atrophy which was due primarily to a decrease in individual fiber area as opposed to fiber loss. Histochemical maturation of the EDL was severely affected by neonatal denervation during the first three postnatal weeks. By 21 days, two extrafusal fiber types which were both oxidative could be distinguished. One type was highly atrophied and resembled an immature fiber exhibiting myosin ATPase staining at both acid and alkaline preincubation conditions, whereas another type was less atrophied and showed myosin ATPase staining resembling fast-twitch (type IIA) fibers. These findings emphasize the importance of an intact nerve supply in determining the phenotypic expression of skeletal muscle, and point to the early postnatal period as a critical stage in fiber type differentiation. PMID- 2970448 TI - Sibling response to chronic childhood disorders: research perspectives and practice implications. PMID- 2970449 TI - Echocardiographic findings in 103 cats with hyperthyroidism. AB - Using M-mode echocardiography, cardiac abnormalities were studied in 103 cats with untreated hyperthyroidism. In addition, follow-up echocardiography was performed on 24 of these cats to assess the long-term (4 to 21 months) effect of treatment (thyroidectomy or radioiodine) on thyrotoxic cardiac disease. The most common echocardiographic abnormality in the 103 untreated hyperthyroid cats was hypertrophy of the left ventricular caudal wall (71.9%). Hypertrophy of the interventricular septum also was documented in 39.8% of the 103 cats. Other abnormalities included high values for left atrial diameter (70.0%), aortic root diameter (18.5%), and left ventricular diameter at end diastole (45.6%). In some of these cats, indices of contractility were enhanced; in 21.4% and 14.6% of the cats, values for shortening fraction and velocity of circumferential fiber shortening, respectively, were greater than those values measured in clinically normal cats. After treatment of the hyperthyroidism, left ventricular hypertrophy resolved or improved in many of the cats, as indicated by decreases in left ventricular caudal wall and interventricular septum thicknesses. Hyperdynamic wall motion resolved in all cats after treatment, as evidenced by consistent decreases in shortening fraction and velocity of circumferential fiber shortening. Despite these improvements, some cats had one or more persistently abnormal echocardiographic values after treatment. These results suggested that in cats, hyperthyroidism commonly is associated with largely reversible cardiomyopathy. In those cats in which cardiomyopathy persists or worsens after treatment, underlying primary cardiomyopathy or thyroid hormone-induced cardiac structural damage may exist. PMID- 2970450 TI - Immune-mediated polysynovitis in four foals. AB - The deposition of immune complexes in the synovial membrane resulted in polysynovitis in 4 foals. All 4 foals had an infection at a site other than the joints. The polysynovitis was characterized by marked effusions of affected joints and joint stiffness. Bacterial and mycoplasmal cultures of the joints did not yield growth. Staining of synovial membrane biopsy specimens with fluorescein labeled anti-equine IgG revealed immune complexes in the synovial membrane. Immune-mediated polysynovitis might develop in foals with bacterial infections. We propose that deposition of immunoglobulin in the synovial membrane of the affected foals was caused by an increase in circulating immune complexes formed as a result of the primary disease processes. PMID- 2970451 TI - Polycythemia vera in a cat with cardiac hypertrophy. AB - Polycythemia vera, a rare and poorly documented disease in cats, was diagnosed in a 4-year-old domestic shorthair cat admitted because of seizures. The diagnosis was made on the basis of high PCV, normal serum erythropoietin concentration (as determined by bioassay, using rabbit bone marrow cells), and elimination of secondary polycythemia as a diagnosis. Cardiac hypertrophy, which might have been secondary to blood hyperviscosity, also was diagnosed. The cat has been treated by periodic phlebotomy and has been without clinical signs of disease for more than 20 months. PMID- 2970452 TI - In-vitro activity of trospectomycin sulphate against Mycoplasma and Ureaplasma species isolated from humans. AB - Trospectomycin sulphate, a novel analogue of spectinomycin, was compared to spectinomycin, tetracycline, lincosamide, macrolide, quinolone and naphthalenic ansamycin-class antibiotics for in-vitro activity against Mycoplasma pneumoniae, M. hominis and Ureaplasma urealyticum. MIC determinations were conducted by the agar dilution method using either SP-4 (Mycoplasma spp.) or A8 (U. urealyticum) agars. Trospectomycin compared favorably with tetracycline for all the strains of M. pneumoniae, M. hominis or U. urealyticum tested. Ciprofloxacin was the most active of the newer quinolones tested. Trospectomycin was as active as this quinolone antibiotic, or more active, in vitro. Other antibiotics such as the macrolides and lincosamides were highly active against one or more of the three mycoplasma groups tested but none was active against all three. The results suggest that trospectomycin may be useful in the treatment of mycoplasma-induced respiratory or genital tract infections in man. PMID- 2970453 TI - The understanding and use of interpersonal gestures by autistic and Down's syndrome children. AB - Autistic adolescents with mild, moderate, and severe degrees of mental retardation, Down's syndrome adolescents, and clinically normal 4-, 5-, and 6 year-old children were compared in their ability to understand a set of simple instrumental gestures. Almost all gestures were perfectly understood, that is, correctly responded to, by normal children from age 5 onwards, and by all the handicapped groups, regardless of diagnosis or degree of retardation. However, the ability to initiate such gestures on verbal request was generally less good, especially in the less able autistic groups. The same subjects were unobtrusively observed in the playground and during mealtime at their schools. Peer interactions were least frequent in the autistic subjects, regardless of degree of mental retardation. However, relative to interaction frequency, the autistic group used nonverbal instrumental gestures as a means of communication to the same extent as the other groups. Unlike Down's syndrome adolescents, or normal preschool children, no autistic adolescent ever used expressive gestures. PMID- 2970454 TI - A comparison of potentially pathologic factors in European children with autism, Down's syndrome, and multiple physical handicaps. PMID- 2970456 TI - Drug profiling by computed electroencephalography and brain maps, with special consideration of sertraline and its psychometric effects. AB - The five main psychopharmacological drug groups are reviewed in respect of their effects on the pharmaco-electroencephalogram (pharmaco-EEG), with special consideration given to drug profiling by topographic brain mapping. In addition, the CNS effects of sertraline (100, 200, and 400 mg), zimelidine (100 mg), and placebo were studied. The pharmaco-EEG profiles of both sertraline (100 mg) and zimelidine were indicative of vigilance-enhancement seen after desipramine-type antidepressants; with higher sertraline dosages there was a change toward the sedation associated with imipramine-type antidepressants. Both sertraline (100 mg) and zimelidine improved psychometric performance, although some deterioration occurred with higher sertraline (200 and 400 mg) doses. Psychophysiological variables showed dose-dependent changes in CNS activation and pupillary dilatation. Adverse effects were minimal with both sertraline (100 mg) and zimelidine, while nausea and vomiting increased with sertraline 200 and 400 mg. PMID- 2970455 TI - Novel activation of araC expression and a DNA site required for araC autoregulation in Escherichia coli B/r. AB - Mutations in the araC gene have been isolated which alter both the activator and autoregulatory functions of AraC protein (L.G. Cass and G. Wilcox, J. Bacteriol. 166:892-900, 1986). In this study, the effect of each araC mutation on autoregulation was characterized in vivo and in vitro in the presence of L arabinose. The effect of L-arabinose in some of these araC mutants revealed a novel activation of araC expression which was not observed in the araC+ cell. Experiments were therefore focused on understanding the mechanism of this novel activation. We describe a systematic analysis of the effect of mutations within the known regulatory binding sites for araBAD and araC transcription on araC expression. Our results suggest that the novel activation of araC expression requires the AraC activator-binding site, araI, and the cyclic AMP receptor protein-cyclic AMP complex-binding site. We also found that in the absence of L arabinose, the araI site was required for maximal autoregulation by the wild-type AraC protein. PMID- 2970457 TI - Reaction cycle of solubilized monomeric Ca2+-ATPase of sarcoplasmic reticulum is the same as that of the membrane form. AB - Monomeric Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum dispersed in Triton X-100 is stoichiometrically phosphorylated from Pi in a Ca2+-depleted medium containing dimethyl sulfoxide and catalyzes efficient (80%) phosphoryl transfer to ADP following a jump in water activity in the presence of Ca2+. The Ca2+ concentration dependence of ATP synthesis was sigmoidal (nH = 1.7) and in the millimolar range (K0.5 = 0.3 mM), indicating the involvement of at least two low affinity Ca2+ binding sites. These results, taken together with the properties of the monomer in the forward direction of catalysis, show that the catalytic cycle of the detergent-solubilized monomer is essentially the same as that of the membrane enzyme. The substrate and ion specificity of the catalytic intermediates suggest that the monomer is capable of coupled vectorial transport of Ca2+. PMID- 2970458 TI - ATP regulation of sarcoplasmic reticulum Ca2+-ATPase. Metal-free ATP and 8-bromo ATP bind with high affinity to the catalytic site of phosphorylated ATPase and accelerate dephosphorylation. AB - To localize and characterize the regulatory nucleotide site of skeletal muscle sarcoplasmic reticulum Ca2+-ATPase, we have investigated the effects of ADP, ATP, and analogues of these nucleotides on the rate of dephosphorylation of both native ATPase and ATPase modified with fluorescein 5'-isothiocyanate (FITC), a reagent which hinders access of nucleotides to the ATPase catalytic site without affecting phosphorylation from Pi. Dephosphorylation of the phosphoenzyme formed from Pi was monitored by rapid filtration or stopped-flow fluorescence, mostly at 20 degrees C, pH 6.0, and in the absence of potassium. Fluorescence measurements were made possible through the use of 8-bromo-ATP, which selectively quenched certain tryptophan residues of the ATPase, thereby allowing the intrinsic fluorescence changes associated with dephosphorylation to be measured in the presence of bound nucleotide. ATP, 8-bromo-ATP, and trinitrophenyladenosine diand triphosphate, but not ADP, enhanced the rate of dephosphorylation of native ATPase 2-3-fold when added in the absence of divalent cations. Millimolar concentrations of Mg2+ eliminated the accelerating effects. Acceleration in the absence of Mg2+ was observed at relatively low concentrations of ATP and 8-bromo ATP (0.01-0.1 mM) and binding of metal-free ATP and ADP, but not Mg.ATP, to the phosphoenzyme in this concentration range was demonstrated directly. Modification of the ATPase with FITC blocked nucleotide binding in the submillimolar concentration range and eliminated the nucleotide-induced acceleration of dephosphorylation. These results show that dephosphorylation, under these conditions, is regulated by ATP but not by Mg.ATP or ADP, and that the catalytic site is the locus of this "regulatory" ATP binding site. PMID- 2970459 TI - Biosynthesis of blood group i-active polylactosaminoglycans. Partial purification and properties of an UDP-GlcNAc:N-acetyllactosaminide beta 1----3-N acetylglucosaminyltransferase from Novikoff tumor cell ascites fluid. AB - An N-acetylglucosaminyltransferase has been partially purified from Novikoff tumor cell ascites fluid by affinity chromatography on concanavalin A-Sepharose. The enzyme was obtained in a highly concentrated form after lyophilization. The enzyme appeared to be highly specific for acceptor oligosaccharides and glycoproteins carrying a terminal Gal beta 1----4GlcNAc beta 1----R unit. Characterization of products formed by the enzyme in vitro by methylation analysis and 1H NMR spectroscopy revealed that the enzyme catalyzed the formation of a GlcNAc beta 1----3Gal beta 1----4GlcNAc beta-R sequence. The enzyme therefore could be described as an UDP-GlcNAc:Gal beta 1----4GlcNAc beta-R beta 1 ---3-N-acetylglucosaminyltransferase. Acceptor specificity studies with oligosaccharides that form part of N-glycans revealed that the presence of a Gal beta 1----4GlcNAc beta 1----2(Gal beta 1----4GlcNAc beta 1----6)Man pentasaccharide in the acceptor structure is a requirement for optimal activity. Studies on the branch specificity of the enzyme showed that the branches of this pentasaccharide structure, when contained in tri- and tetraantennary oligosaccharides, are highly preferred over other branches for attachment of the 1st and 2nd mol of GlcNAc into the acceptor molecule. The enzyme also showed activity toward oligosaccharides related to blood group I- and i-active polylactosaminoglycans. In addition the enzyme together with calf thymus UDP Gal:GlcNAc beta-R beta 1----4-galactosyltransferase was capable of catalyzing the synthesis of a series of oligomers of N-acetyllactosamine. Competition studies revealed that all acceptors were acted upon by a single enzyme species. It is concluded that the N-acetylglucosaminyltransferase functions in both the initiation and the elongation of polylactosaminoglycan chains of N-glycoproteins and possibly other glycoconjugates. PMID- 2970460 TI - Crystal violet as an uncoupler of oxidative phosphorylation in rat liver mitochondria. AB - Crystal violet exhibited characteristics of an uncoupler of oxidative phosphorylation, i.e. it released respiratory control, hindered ATP synthesis, enhanced ATPase activity, and produced swelling of isolated rat liver mitochondria. Maximal stimulation of respiration, ATPase activity, and swelling was observed at a concentration of 40 microM. The inhibition of State 3 respiration by oligomycin was released by crystal violet. High concentrations of crystal violet inhibited mitochondrial respiration. The uncoupling effect of crystal violet required inorganic phosphate and was abolished by N ethylmaleimide. The adenine nucleotides ADP and ATP protected mitochondria from uncoupling by the dye. The dye taken up by mitochondria was released into the incubation medium on induction of uncoupling. In the absence of phosphate, the dye did not cause uncoupling, but its retention was much greater than in the presence of phosphate. Crystal violet is suggested to induce uncoupling by acting on the membrane, rather than by its electrophoretic transfer into the mitochondria. PMID- 2970462 TI - Insulin receptors with defective tyrosine kinase inhibit normal receptor function at the level of substrate phosphorylation. AB - Rat 1 fibroblasts have been transfected with the cDNA encoding a kinase-defective mutant human insulin receptor (A/K1018). Expression of this cDNA results in a receptor that is not only biologically inactive but also inhibits normal insulin action through the normal endogenous rat receptors in this fibroblast line (McClain, D. A., Maegawa, H., Lee, J., Dull, T. J., Ullrich, A., and Olefsky, J. M. (1987) J. Biol. Chem. 262, 14663-14671). We have investigated the mechanism of this inhibition and show that: 1) rat receptors are expressed at normal to increased levels in two cell lines which also express A/K1018 receptors at low (A/K1018-A, 5700 total receptors) or high (A/K1018-B, 2.2 x 10(5) total receptors) levels. 2) The rat receptors in the A/K1018 lines can be normally autophosphorylated under the control of insulin in vitro. 3) A/K1018 receptors do not inhibit the kinase activity of normal receptors when mixed together in vitro. 4) In intact A/K1018-B cells, the ability of insulin to stimulate autophosphorylation of the rat receptor is unimpaired; furthermore, the autophosphorylated rat receptor becomes normally activated as a tyrosine kinase. 5) The expression of receptors for insulin-like growth factor I and stimulation of hexose uptake mediated by this receptor are unaffected in cells expressing inhibitory A/K1018 receptors. 6) Expression of the A/K1018 receptor inhibits insulin-stimulated phosphorylation of two endogenous protein substrates (pp220 and pp170) by the native rat receptors. We conclude that the inhibition of insulin action seen in the A/K1018 cells is not mediated at the levels of native receptor expression or activation, nor is the effector (hexose uptake) mechanism affected by the A/K1018 receptors. The expression of this kinase-defective receptor does, however, inhibit the phosphorylation of substrate molecules by the normally activated endogenous rat receptors. PMID- 2970461 TI - The secretion of atrial natriuretic factor-(99-126) by cultured cardiac myocytes is regulated by glucocorticoids. AB - Atrial natriuretic factor (ANF) is stored in mammalian atria primarily as ANF-(1 126), the precursor to the known circulating form of the hormone ANF-(99-126). When primary cultures of atrial myocytes were maintained in a complete serum-free medium, they contained and secreted an ANF-(1-126)-like peptide. The addition of dexamethasone to the culture medium, however, resulted in the secretion of a molecule with chromatographic characteristics identical to ANF-(99-126), although the intracellular storage form of ANF was unchanged. Radiosequencing and amino acid analysis confirmed that the cultures maintained in dexamethasone secreted authentic ANF-(99-126). Chronic exposure of the cells to dexamethasone also resulted in a significant increase in the quantity of immunoreactive ANF both contained and secreted by the cultures. Dexamethasone stimulated ANF processing and secretion by atrial cultures in a dose-dependent manner, with an approximate EC50 of 10 nM. This stimulation could be reversed by removing the glucocorticoid from the culture medium. ANF processing was also stimulated by the specific glucocorticoid receptor agonist RU 28362, and both DEX- and RU 28362-stimulated ANF processing was inhibited by the specific glucocorticoid receptor antagonist RU 38486. Ventricular cells, which possess few granules and release ANF in a constitutive fashion, were also capable of processing ANF in a glucocorticoid dependent fashion. Medium freshly removed from atrial cultures did not convert ANF-(1-126) to ANF-(99-126) nor was exogenous ANF-(1-126) efficiently processed when added to the medium of actively processing cultures. These results indicate that the post-translational processing of ANF-(1-126) to ANF-(99-126) likely occurs within or in close association with the cardiac myocytes and is not dependent on the presence of large quantities of secretory granules. Furthermore, it is apparent that both the expression and the post-translational processing of ANF by cultured cardiac myocytes is specifically regulated by glucocorticoids. PMID- 2970463 TI - Purification and characterization of the Mg2+-ATPase from rabbit skeletal muscle transverse tubule. AB - Transverse tubule membranes isolated from rabbit fast skeletal muscle contain a very active Mg2+-ATPase (ATP phosphohydrolase, EC 3.6.1.3). This enzyme is very sensitive to inactivation by most detergents. However, after solubilization with either lysolecithin or digitonin, the Mg2+-ATPase can be purified in active form. Using a combination of selective solubilization followed by lectin affinity chromatography, ion-exchange chromatography, and native gel electrophoresis, the Mg2+-ATPase has been purified to near homogeneity. A prominent band with molecular mass of 105 kDa is observed when the purified protein is analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified 105-kDa Mg2+-ATPase protein is not structurally similar to the sarcoplasmic reticulum Ca2+-ATPase protein, as evidenced by very different cyanogen bromide peptide maps and amino acid compositions. The structural dissimilarities are complemented by functional differences observed between the Ca2+- and Mg2+-ATPases, including differential susceptibility to proteases, chemical modification reagents, and inactivation by fluorescein isothiocyanate and vanadate. All these data taken together demonstrate that the Mg2+-ATPase is a unique protein with little, if any, structural similarity to the sarcoplasmic reticulum Ca2+-ATPase or to other related enzymes such as mammalian kidney (Na,K)-ATPase or gastric mucosal (H,K) ATPase. PMID- 2970464 TI - Purification and characterization of kinesin from bovine adrenal medulla. AB - Kinesin was purified from bovine adrenal medulla. The sedimentation coefficient was 8.8 S. Sedimentation equilibrium ultracentrifugation studies showed the molecular weight of kinesin to be 300,000. The calculated axial ratio was 1:16. The Stokes radius was estimated to be 8.9 nm by gel filtration. Circular dichroism showed the alpha-helix content to be about 50%. Purified kinesin preparation contained a major polypeptide with a molecular weight of 120,000 and minor ones with molecular weights of 71,000, 68,000, and 65,000. Bovine adrenal kinesin had an ATPase activity which was stimulated severalfold by microtubules to a specific activity of about 0.1 mumol/min.mg. Kinesin molecules adsorbed to a glass slide promoted the movement of microtubules on the glass surface at a rate of about 0.5 micron/s. Immunostaining of EBTr (bovine embryonic trachea fibroblast) cells and bovine adrenal chromaffin cells in interphase with an affinity-purified antibody against the major polypeptide of kinesin showed that some kinesin was located on microtubules and the rest distributed throughout the cytoplasm in a diffuse manner. EBTr cells in mitotic phase gave a staining pattern showing that kinesin was present throughout the cytoplasm with higher concentration in the region of mitotic apparatus. PMID- 2970465 TI - Novel subunit in C4b-binding protein required for protein S binding. AB - C4b-binding protein (C4BP) is a multimeric protein with regulatory functions in the complement system. It also interacts with vitamin K-dependent protein S, which is involved in the regulation of the coagulation system. It has been demonstrated that C4BP consists of seven disulfide-linked, identical 70-kDa subunits, which are arranged to give the molecule a spider-like structure. We now have evidence for the presence of a new subunit in C4BP. On sodium dodecyl sulfate-poly-acrylamide gel electrophoresis it appears as a weakly stainable band with a molecular weight of approximately 45,000. The subunit was isolated by gel filtration in 6 M guanidine hydrochloride of reduced and carboxymethylated C4BP. Its amino-terminal sequence is distinct from previously known protein sequences. The stoichiometry of 45- to 70-kDa subunits was estimated to be 1:9, indicating the presence of one 45-kDa subunit per C4BP molecule. The new subunit was demonstrated to be a disulfide-linked component of the central core of C4BP. It was sensitive to proteolysis by chymotrypsin, and when cleaved the protein S binding ability of C4BP was lost. With protein S bound to C4BP, the 45-kDa subunit was protected from degradation by chymotrypsin, and the protein S binding site remained intact. These data suggest that the new subunit is directly involved in protein S binding. PMID- 2970466 TI - Adhesion defective BHK cell mutant has cell surface heparan sulfate proteoglycan of altered properties. AB - In the light of accumulating data that implicate cell surface heparan sulfate proteoglycans (HSPGs) with a role in cell interactions with extracellular matrix molecules such as fibronectin, we have compared the properties of these molecules in wild-type BHK cells and an adhesion-defective ricin-resistant mutant (RicR14). Our results showed that the mutant, unlike BHK cells, cannot form focal adhesions when adherent to planar substrates in the presence of serum. Furthermore, while both cell lines possess similar amounts of cell surface HSPG with hydrophobic properties, that of RicR14 cells had decreased sulfation, reduced affinity for fibronectin and decreased half-life on the cell surface when compared to the normal counterpart. Our conclusions based on this data are that these altered properties may, in part, account for the adhesion defect in the ricin-resistant mutant. Whether this results from the known alteration in assembly of N-linked glycans affecting the carbohydrate chains on the proteoglycan or some other combination of factors is discussed. PMID- 2970467 TI - Structure, biosynthesis, and function of the hexose transporter in Chinese hamster ovary cells deficient in N-acetylglucosaminyltransferase 1 activity. AB - We have used a Chinese hamster ovary cell line deficient in N acetylglucosaminyltransferase 1 activity (Lec1) to study the effects of altered asparagine-linked oligosaccharides on the structure, biosynthesis, and function of glucose transporter protein. Immunoblots of membranes of Lec1 cells show a glucose transporter protein of Mr 40,000, whereas membranes of wild-type (WT) cells contain a broadly migrating Mr 55,000 form similar to that observed in several other mammalian tissues. The total content of immunoreactive glucose transporters in Lec1 cells is 3.5-fold greater than that of WT cells. Digestion with endoglycosidases, treatment with inhibitors of glycosylation, and interactions with agarose-bound lectins demonstrate that glucose transporters of both cell lines derive from a similar Mr 38,000 core polypeptide and that both contain asparagine-linked oligosaccharide. Transporters in Lec1 cells contain primarily "undecorated" but "trimmed" mannose-type asparagine-linked oligosaccharides, while the protein in WT cells contains a mixture of "decorated" and "trimmed" asparagine-linked oligosaccharides. Biosynthetic and turnover studies demonstrate that Lec1 cells, in contrast to WT cells, are unable fully to process the core asparagine-linked oligosaccharides of maturing glucose transporters. When radiolabeled in methionine-deficient medium both Lec1 and WT cells show similar rates of synthesis and turnover of glucose transporter proteins. It should be noted, however, that starvation for a critical amino acid may alter the ability of the cell to synthesize or degrade proteins. The abilities of Lec1 and WT cells to transport hexoses and to interact with the inhibitor cytochalasin B are very similar. The results indicate that, although altered asparagine-linked glycosylation can affect the content and biogenesis of glucose transporters, these changes do not greatly modify cellular hexose uptake. The possibility that alterations in asparagine-linked glycosylation may change the cell surface localization or acquisition of a "functional conformation" of the glucose transporter is also suggested. PMID- 2970468 TI - Phosphorylation of ankyrin down-regulates its cooperative interaction with spectrin and protein 3. AB - Ankyrin mediates the primary attachment between beta spectrin and protein 3. Ankyrin and spectrin interact in a positively cooperative fashion such that ankyrin binding increases the extent of spectrin tetramer and oligomer formation (Giorgi and Morrow: submitted, 1988). This cooperative interaction is enhanced by the cytoplasmic domain of protein 3, which is prepared as a 45-41-kDa fragment generated by chymotryptic digestion of erythrocyte membranes. Using sensitive isotope-ratio methods and nondenaturing PAGE, we now demonstrate directly (1) the enhanced affinity of ankyrin for spectrin oligomers compared to spectrin dimers; (2) a selective stimulation of the affinity of ankyrin for spectrin oligomer by the 43-kDa cytoplasmic domain of protein 3; and (3) a selective reduction in the affinity of ankyrin for spectrin tetramer and oligomer after its phosphorylation by the erythrocyte cAMP-independent membrane kinase. The phosphorylation of ankyrin does not affect its binding to spectrin dimer. Ankyrin also enhances the rate of interconversion between dimer-tetramer-oligomer by 2-3-fold at 30 degrees C, and in the presence of the 43-kDa fragment, ankyrin stimulates the rate of oligomer interconversions by nearly 40-fold at this temperature. These results demonstrate a long-range cooperative interaction between an integral membrane protein and the peripheral cytoskeleton and indicate that this linkage may be regulated by covalent protein phosphorylation. Such interactions may be of general importance in nonerythroid cells. PMID- 2970469 TI - Biospecific sorption of cyclodextrin glucosyltransferase on physically modified starch. PMID- 2970470 TI - Gamma-atrial natriuretic polypeptide (gamma ANP)-derived peptides in human plasma: cosecretion of N-terminal gamma ANP fragment and alpha ANP. AB - Using RIAs for the N- and C-terminal fragments of the human atrial natriuretic polypeptide (ANP) precursor gamma ANP, that is gamma ANP-(1-25), and alpha ANP [gamma ANP-(99-126)], we studied the secretion of gamma ANP-derived peptides from the heart in normal subjects and patients with heart disease, chronic renal failure, and cirrhosis. We detected gamma ANP-(1-25)-like immunoreactivity (-LI) in plasma from normal subjects (n = 17) in considerable amounts [mean, 510 +/- 62 (+/- SE) pg/mL (174 +/- 21 pmol/L)]; the mean plasma alpha ANP-LI level at the same time in these subjects was 32.8 +/- 4.4 pg/mL (10.7 +/- 1.4 pmol/L). Gel permeation chromatographic analysis of plasma samples from normal subjects and patients with heart disease and chronic renal failure revealed two major components; one was alpha ANP, and the other was the 10K N-terminal gamma ANP fragment (N-peptide) resulting from the removal of alpha ANP (3K) from gamma ANP (13K). In addition, gamma ANP (13K), which possessed both gamma ANP-(1-25)-LI and alpha ANP-LI, and beta ANP, an antiparallel dimer of alpha ANP, were detected in some patients as minor components. A significant positive correlation between plasma levels of the N-terminal gamma ANP fragment and alpha ANP (P less than 0.01) and almost equal step-ups in the coronary sinus plasma levels of the N terminal gamma ANP fragment and alpha ANP suggest that they are cosecreted in equimolar amounts. The high molar ratio of plasma gamma ANP-(1-25)-LI to alpha ANP-LI (17.4 +/- 1.4) in normal subjects and the significantly higher ratio in patients with chronic renal failure (36.9 +/- 7.1; P less than 0.01) suggest the slower clearance of the N-terminal gamma ANP fragment than alpha ANP and a role for the kidney in its degradation. Since the molar ratio of plasma gamma ANP-(1 25)-LI to alpha ANP-LI in patients with cirrhosis (20.7 +/- 2.7) was similar to that in normal subjects, it is unlikely that the N-terminal gamma ANP fragment is metabolized by the liver. In patients with heart disease, plasma gamma ANP-(1-25) LI and alpha ANP-LI levels were higher in those with cardiac decompensation and were positively correlated with right atrial pressure, pulmonary arterial pressure, and pulmonary capillary wedge pressure, indicating cosecretion of the N terminal gamma ANP fragment and alpha ANP in response to atrial stretch.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970471 TI - Plasma atrial natriuretic hormone levels in patients with the syndrome of inappropriate antidiuretic hormone secretion. AB - This study explored whether atrial natriuretic hormone (ANH) might be involved in the escape from salt and water retention that occurs in patients with the syndrome of inappropriate antidiuretic hormone secretion (SIADH). Sixteen patients with low serum Na+ concentrations [123 +/- 1 (+/- SE) mmol/L] were studied. Each patient excreted urine that was hyperosmolar (mean, 391 +/- 4 mosmol/kg) in relation to serum osmolality (mean, 258 +/- 4 mosmol/kg). Sodium excretion (81 +/- 20 mmol/L) also was inappropriate to the low serum Na+ level. The probable causes of SIADH were head trauma (4), pneumonia (5), lung cancer (3), and chlorpropamide therapy (4). In the nontumor patients, plasma and/or urinary vasopressin (AVP) concentrations were in the normal range, but inappropriate for serum osmolality. Urinary AVP values of 50 pg/mL or more (greater than 46 pmol/L) were found in the three tumor patients. The mean plasma ANH concentration was 6-fold higher than that in normal subjects [296 +/- 51 vs. 51 +/- 13 pg/mL (100 +/- 20 vs. 17 +/- 4 pmol/L); P less than 0.01]. Six SIADH patients were studied again after brief (1-3 days) water restriction. Although serum osmolality increased in each, their plasma AVP concentrations decreased very little, and urinary AVP excretion and plasma ANH did not change. These results indicate that plasma ANH levels are markedly increased in patients with SIADH. Their increased ANH secretion may antagonize water retention resulting from the inappropriate AVP secretion. PMID- 2970472 TI - Selective loss of CD4+ CD45R+ T cells in peripheral blood of multiple myeloma patients. AB - The phenotypic distribution of T-lymphocyte subsets in peripheral blood from multiple myeloma (MM) patients shows a reduced proportion of CD4+ cells and a normal proportion of CD8+ cells. The decrease in CD4+ cells could be due to a random process, with all types of CD4+ cells being equally affected, or it could reflect a nonrandom process with selected subsets preferentially reduced. In order to distinguish between these possibilities, double immunofluorescence analysis was performed on blood samples from patients with MM, patients with monoclonal gammopathy of unknown significance (MGUS), and age-matched normal donors, using monoclonal anti-CD4 or anti-CD8 paired with antibodies to the common leukocyte marker Lp220 (CD45R) or 4B4 (CDw29). Normal peripheral blood lymphocytes (PBL) include two phenotypically and functionally distinct CD4+-cell subsets, identified as CD4+ Lp220+ 4B4- and CD4+ Lp220-4B4+, whereas the majority of CD8+ cells expresses Lp220 (70-85%). MM patients had a highly significant selective reduction of the CD4+ Lp220+ subset compared with normal controls (P less than 0.001). Although the percentage of CD4+ Lp220- cells was also reduced in some MM patients relative to normal donors, most of MM patients had an elevated Lp220-/Lp220+ ratio of CD4+ cells (P less than 0.001). The proportion of the two CD8+ subsets was also markedly abnormal. In the set of patients studied the abnormalities within the CD4+ and CD8+ lymphocytes were exclusive to patients with MM since patients with MGUS had normal proportion of CD4+ and CD8+ subsets. PMID- 2970473 TI - Differentiation of human B lymphocyte subpopulations induced by an alloreactive helper T-cell clone. AB - We have used cloned alloreactive helper T cells to determine if direct T cell-B cell interaction can induce differentiation of human peripheral blood B cells which do not respond to pokeweed mitogen (PWM). T-cell clone 2F8 was derived from a one-way mixed lymphocyte reaction. 2F8 cells are T3+T4+T8-IL-2R+ and proliferate in response to irradiated stimulator cells, but not autologous cells, in the absence of exogenous interleukin-2. 2F8 cells provide allospecific help for polyclonal proliferation and differentiation of B cells in the absence of any other stimulus. The magnitude of this response is comparable to that of the response of the same B cells to PWM and fresh autologous T cells. 2F8 cells could also provide nonspecific help for unrelated donor B cells in the presence of PWM, with no requirement for costimulation by irradiated stimulator cells. Allospecific stimulation of B cells was completely inhibited by antibodies to class II major histocompatibility complex (MHC) framework determinants and was abrogated by 1000-rad irradiation. Cloned 2F8 T cells stimulated differentiation of both small, high-density B cells and larger B cells, generating up to 30% plasma cells with either fraction. B cells forming rosettes with mouse erythrocytes were also induced to differentiate by the helper T cell clone. As found previously, neither small, high-density B cells nor mouse rosette+ B cells responded well to PWM. Direct interaction with allospecific T cells induces differentiation of a broader spectrum of B cells than soluble growth and differentiation factors in conjunction with polyclonal activators such as PWM and protein A containing staphylococci. PMID- 2970475 TI - Retinoids and acne. AB - Retinoids, chemicals that have vitamin A activity, have become important therapeutic agents for a variety of cutaneous disorders, including acne. This paper reviews the history of retinoid use in acne, including retinol, topical tretinoin, topical and systemic isotretinoin, systemic etretinate, and new investigational retinoid molecules. PMID- 2970476 TI - Systemic retinoid therapy for psoriasis. AB - Retinoids are potent therapuetic agents that have been found to be effective in a variety of skin diseases. They are of benefit in various forms of severe psoriasis. With severe plaque psoriasis, they are used most effectively in combination with other forms of therapy, such as phototherapy. With generalized pustular psoriasis, they are effective monotherapy and are frequently helpful for the control of exfoliative psoriasis. A variety of new retinoid analogs have been studied in clinical investigations. This article discusses important aspects of the use of etretinate, isotretinoin, acitretin, and arotinoid ethyl ester in the treatment of severe forms of psoriasis. PMID- 2970477 TI - Effects of retinoids in bone. AB - Skeletal effects of retinoids on the spine were studied in two clinical trials. In the first study, spinal radiographs of 96 patients who had been treated with isotretinoin for 4 to 9 months were reviewed. The average age of these patients was 25 years, and during treatment or within 2 1/2 years after the end of treatment, 26% of the patients showed progressive formation of small bony spurs consisting of tiny horizontal excrescences that arose at the anterior margin of one or more vertebral bodies adjacent to the intervertebral disk. In a second study, the radiographs of 241 patients with psoriasis who were treated continually for 1 to 2 years with acitretin were examined. Many of these patients had abnormal radiographs at the start of therapy. These preexisting conditions included psoriatic arthritis, degenerative arthritis, and diffuse idiopathic skeletal hyperostosis. Five percent of the patients showed progression of their abnormalities during the study. The difference in the rate of spur formation in the two groups may be due to multiple factors and not simply to retinoid therapy. Because of the extensive amount of preexisting disease in the psoriasis group compared with the relatively normal appearance of the spine in the isotretinoin group, the underlying disease process may be more important than the retinoid therapy. The development of the spinal spurs was not associated with specific clinical symptoms. Since there was no control group, it is unknown whether the spurs would have developed or progressed in the absence of retinoid therapy. PMID- 2970478 TI - Subacute cutaneous lupus erythematosus presenting as erythroderma. AB - We report the first case of subacute cutaneous lupus erythematosus presenting with exfoliative erythroderma. Although our patient had uniquely extensive skin disease, she had the clinical, histologic, serologic, and immunogenetic features of subacute cutaneous lupus erythematosus. Complete remission was obtained with a short course of systemic corticosteroids and long-term hydroxychloroquine therapy. We briefly discuss the usual features of subacute cutaneous lupus erythematosus. PMID- 2970479 TI - Gentamicin: systemic exposure to a contact allergen. AB - A case is presented of an allergic dermatitis provoked by intravenous gentamicin in a patient previously sensitized by topical medications. Patch tests confirmed hypersensitivity to gentamicin and neomycin. The nature of reactions to contact allergens given systemically and the nature of cross-reactions between aminoglycoside antibiotics are reviewed. PMID- 2970474 TI - Pathway for the communication between the ATPase and actin sites in myosin. PMID- 2970480 TI - A national survey of sealant use by pediatric dentists. AB - A national survey of pediatric dentists was undertaken to determine their level of sealant use and the factors that affected level of use. A twelve-page mailed questionnaire was completed by 329 practitioners. Decision-making factors, attitudes and knowledge about sealants, sealant awareness, characteristics of sealant use, undergraduate dental school experience with sealants, auxiliary factors, impact of insurance, characteristics of dental practice, interaction with colleagues, and characteristics of patient populations were explored as they related to level of sealant use. Variables significantly associated with sealant use included: availability of insurance, ability to delegate sealant procedures, and patient income and acceptance. Frequent users were more knowledgeable about sealant issues and were more likely to be consulted by their peers. PMID- 2970481 TI - The curved-bristle toothbrush: an aid for the handicapped population. AB - This study was undertaken to assess the comparative effectiveness of two toothbrushes in the removal of plaque in an institutionalized, handicapped population. The two methods were: a conventional toothbrush using the Modified Stillman Method; and a curved-bristle toothbrush using the scrubbing technique recommended by the manufacturer. Subjects selected were profoundly retarded individuals, incapable of self-brushing skills. According to random selection, subjects' teeth were brushed by trained personnel with either the curved bristle or the conventional toothbrush. Results demonstrated that the curved-bristle toothbrush was as effective as the conventional toothbrush when used by trained personnel. PMID- 2970482 TI - Marginal leakage associated with three posterior restorative materials. PMID- 2970483 TI - [Oral symptom and sign in AIDS: finding among 23 homosexual men]. PMID- 2970484 TI - Muscle spindles in the digastric muscle of the rabbit. AB - The digastric muscle of the rabbit was studied for the presence of muscle spindles. One of the 19 adult digastric muscles we examined contained in excess of 12 spindles scattered throughout the belly. No spindles were found in the contralateral muscle, or in any of the digastric muscles from other animals. Although jaw depressor muscles of most mammals contain few, if any, muscle spindles, their occasional presence suggests that these muscles have a potential for spindle formation. PMID- 2970485 TI - Preliminary evaluation of a multichannel electrotactile speech processor. AB - Speech discrimination testing, using both open- and closed-set materials, was carried out with four severely to profoundly hearing impaired adults and seven normally hearing subjects to assess performance of a wearable eight-channel electrotactile aid (Tickle Talker). Significant increases in speechtracking rates were noted for all subjects when using the electrotactile aid. After 70 h of training, mean tracking rate in the tactile plus lipreading condition was 55 words per minute (wpm), as compared with 36 wpm for lipreading alone, for the normally hearing group. For the hearing impaired group, the mean tracking rate in the aided condition was 37 wpm, as compared with 24 wpm for lipreading alone, following 35 h of training. Performance scores on Central Institute for the Deaf (CID) everyday sentences, Consonant Nucleus Consonant (CNC) words, and closed-set vowel and consonant identification were significantly improved when using the electrotactile aid. Performance scores, using the aid without lipreading, were well above chance on consonant and vowel identification and on elements of the Minimal Auditory Capabilities Battery. Two hearing impaired subjects have used the device satisfactorily in the home environment. PMID- 2970486 TI - Perception of aperiodicities in synthetically generated voices. AB - The purpose of this study was to investigate univariate relationships between perceived dysphonia and variation in pitch perturbation, amplitude perturbation, and additive noise. A time-domain, pitch-synchronous synthesis technique was used to generate sustained vowels varying in each of the three acoustic dimensions. A panel of trained listeners provided direct magnitude estimates of roughness in the case of the stimuli varying in pitch and amplitude perturbation, and breathiness in the case of the stimuli varying in additive noise. Very strong relationships were found between perceived roughness and either pitch or amplitude perturbation. However, unlike results reported previously for nonspeech stimuli, the subjective quality associated with pitch perturbation was quite different from that associated with amplitude perturbation. Results also showed that perceived roughness was affected not only by the amount of perturbation, but also by the degree of correlation between adjacent pitch or amplitude values. A strong relationship was found between perceived breathiness and signal-to-noise ratio. Contrary to previous findings, there was no interaction between signal-to noise ratio and the amount of high-frequency energy in the periodic component of the stimulus: Stimuli with similar signal-to-noise ratios received similar ratings, regardless of differences in the spectral slope of the periodic component. PMID- 2970487 TI - Suicide and course of illness in major affective disorder. AB - The relationship between a history of attempted suicide and prior course of illness was explored in 87 patients with major affective disorder. Fifty-eight percent of bipolar (n = 67) and 50% of unipolar (n = 20) patients had a history of a suicide attempt. Females were more likely to have made an attempt (67%) than males (42%) and were equally likely to have made a violent attempt. Course of illness and prior history of psychosis were similar in patients with and without a history of an attempt. However, in patients who made an attempt, the severity of the worst attempt was positively correlated with the duration of illness. Greater numbers of prior attempts were associated with greater lethality of the worst attempt. Suicidal ideation for the period of most severe depression in the prior 12 months did not correlate with any measure of lethality of suicide attempt. These data not only suggest the need for further studies of suicide in relationship to the longitudinal course of affective illness, but also the need for continuous reappraisal of suicidal risk in patients with recurrent affective disorders. PMID- 2970488 TI - Failure to detect organic factors in mania. AB - Forty patients from an outpatient lithium clinic were studied who had a clear history of organic abnormalities which predated their affective symptomatology. In the course of reviewing the clinical histories regarding these patients, it was observed that only 37.5% of the patients had ever received a clinical diagnosis of organic affective syndrome. Variables associated with a failure to consider a diagnosis of organic affective syndrome were investigated. PMID- 2970489 TI - Diagnosis and outcome in schizo-affective depression: a replication. AB - Forty-three inpatients with RDC schizo-affective depression were given structured interviews and then followed to 1 year using a design closely resembling that of another recent follow-up of schizo-affective patients. In replication of the earlier study, patients with either 'chronic' or 'mainly schizophrenic' schizo affective disorder had significantly worse outcomes than did patients with nonchronic or 'mainly affective' schizo-affective depression and bipolar patients were significantly more likely to develop manic syndromes. The preceding duration of schizophrenia-like symptoms and a history of schizophrenia-like prodromes appeared to be the most important components of these two distinctions. In both studies, diagnostic subtype was the most robust of various potential outcome predictors. Also in both studies, 'mainly affective' and 'nonchronic' schizo affective patients had outcomes no different from patients with psychotic depression. PMID- 2970490 TI - The clinical picture of the depressed form of puerperal psychosis. AB - The clinical features of 104 patients admitted to a mother and baby unit over a 5 year period with RDC diagnoses of depression or schizoaffective depression were studied by self-ratings, measurements of non-verbal behaviour from standardised videotaped interviews, and observer ratings based on multiple information sources. The hypothesis was tested that they included a large subset which represented the depressed form of a bipolar puerperal psychosis. It was deduced that these illnesses would start early in the puerperium, and would have some specific clinical features. A comparison of patients with onset of depression within 2 weeks of delivery with those with onset during pregnancy or much later in the postnatal period showed a number of significant differences. A discriminant function analysis using serial reclassification of atypical patients refined the early-onset group by the exclusion of about one third of the patients who had an atypical clinical picture; the patients who remained, who are considered to represent the depressed form of puerperal psychosis, showed less anger, less self-rated emotion and more animation than the other depressed patients. PMID- 2970491 TI - Differential changes in areas of social adjustment from depressive episodes through recovery. AB - The findings of the present short-term prospective study of 99 depressed outpatients further support previous cross-sectional observations to the effect that the course of depressive illness is often complicated by fluctuating social disturbances manifested by uneasiness in the work area, by disagreements with colleagues, and by difficulty in maintaining conflict-free relationships with significant others. By contrast, the incapacity to enjoy and use leisure time appeared less related to the symptomatologic variation in depression. Although we favor the hypothesis that impairment in leisure activity may represent a trait marker of depression, the hypothesis of it being a residual complication of repeated depressive episodes cannot be ruled out in view of short follow-up. PMID- 2970492 TI - The Manchester Nurse Rating Scales for the daily simultaneous assessment of depressive and manic ward behaviours. AB - Two nurses' rating scales, the Manchester Nurse Rating Scale for Depression (MNRS D) and the Manchester Nurse Rating Scale for Mania (MNRS-M) are described. The scales were designed for the daily rating of depressive and manic ward behaviours. The inter-rater reliability coefficients (r) for pairs of qualified nurses were found to be 0.790 (MNRS-D) and 0.949 (MNRS-M). All but one of the individual items yielded significant reliability coefficients. The validity coefficients, using psychiatrists' global ratings as the criterion, were 0.533 (MNRS-D) and 0.650 (MNRS-M); somewhat higher correlations were obtained between the MNRS-D and the Montgomery-Asberg Scale and the Beck Depression Inventory, and between the MNRS-M and the Young Mania Scale. The MNRS-D was shown to be sensitive to clinical change following treatment with antidepressant drugs. PMID- 2970493 TI - Efficacy of fluvoxamine in treatment-refractory depression. AB - The efficacy of the potent and selective unicyclic serotonin reuptake inhibitor, fluvoxamine, was evaluated in 38 consecutively admitted depressed patients judged refractory to standard antidepressants using operationalized criteria. Twenty eight patients completed a single-blind protocol involving greater than or equal to 2 weeks of placebo and 4-6 weeks of active fluvoxamine. Eight (29%) were judged responders to fluvoxamine alone, eight (29%) responded to lithium augmentation of fluvoxamine and two (7%) responded to fluvoxamine, lithium and perphenazine. These data suggest that selective and potent serotonin reuptake inhibitors may be effective in patients refractory to generally available antidepressant medications. PMID- 2970494 TI - Age at onset in geriatric depression: relationship to clinical variables. AB - The significance of age at onset of first depressive episode was evaluated in an elderly depressed population. A prospective study of 71 consecutively admitted inpatients with a diagnosis of major unipolar depression examined the relationship between age at onset of illness and several clinical variables. Subjects divided into early-onset (EO) and late-onset (LO) groups, matched for current age, did not significantly differ in terms of symptomatology, cognitive impairment, physical illness, family history or treatment responsivity. These findings do not support a nosologic separation of EO and LO depression in geriatric patients. PMID- 2970495 TI - Bipolar versus unipolar and primary versus secondary affective disorder: which diagnosis takes precedence? AB - The primary versus secondary distinction is often used as a way of subtyping depression. Its applicability to bipolar disorders has been unclear. This report examines the relative primacy of the bipolar versus unipolar distinction as compared to the primary versus secondary distinction in a sample of 955 patients in the NIMH Collaborative Study of the Psychobiology of Depression. These patients are divided into nine groups of the basis of whether they are bipolar I, bipolar II, or unipolar, and whether they are primary, 'pure' secondary, or 'complicated' secondary (i.e. bipolar I primary, bipolar I pure secondary, bipolar I complicated, etc.). Three sets of variables are used to determine the predictive validity of these various subtypes: data concerning age of onset and phenomenology of current episode, outcome, and familial prevalence. In general, data from these three sets of validators suggest that the bipolar distinction takes precedence over the primary versus secondary distinction. Within bipolars, there is little value in further subtyping into primary versus secondary. PMID- 2970497 TI - A family study of rapid-cycling bipolar illness. AB - Twenty-nine out of 195 bipolar/episodic schizoaffective patients were judged to be rapid-cyclers (15%). Twenty-five of the 29 were female (86%). The age corrected morbid risk for major affective disorder was 23.5% in 179 relatives of rapid-cyclers and 31.0% in 189 relatives of matched non-rapid cyclers (chi 2 = 2.6, NS). The prevalence of rapid-cycling itself was also not different in the two groups of relatives. Rapid-cycling thus appears to arise from factors which are separable from the genetic vulnerability to bipolar illness and which do not lead to aggregation within families. PMID- 2970496 TI - Thyrotropin response to thyrotropin-releasing hormone in RDC schizodepressed men. AB - Biological tests may help clarify the relationships of schizoaffective disorder to both major depressive disorder and schizophrenia. Thyrotropin-releasing hormone (TRH), 500 micrograms i.v., was administered to 14 schizodepressed, 23 schizophrenics, 41 unipolar major depressives (all by RDC) and 45 healthy controls, all males 20-67 years old with no significant differences in age, body height or weight. Results showed no differences in maximal delta TSH (dTSH max) amongst schizoaffective depressed, schizophrenia and healthy control groups (10.1 +/- 1.3, 9.2 +/- 1.1, 9.7 +/- 0.8 microU/ml, means +/- SEM respectively). Mean major depressives' dTSH max was lower than in each of the other three groups (6.2 +/- 0.4 microU/ml, P less than 0.01 for all). Utilizing a less than or equal to 5.0 microU/ml cut-off criterion for blunting, the schizodepressed had 36%, schizophrenics 44%, healthy controls 22% and major depressed 59% blunters (P less than 0.05 from other three groups). Schizodepressed patients appeared significantly different from major depressed but closer to schizophrenics (and healthy controls) on the TRH test. PMID- 2970498 TI - Treatment of mixed mania. AB - Mixed mania (i.e., a manic syndrome accompanied by depressive symptoms) and its response to long-term preventive drug treatment was studied as part of a larger NIMH collaborative study. Following recovery from a manic episode, patients received either lithium, imipramine, or the combination of lithium and imipramine for a 2-year period. It was found that patients who had recovered from a mixed manic state were at significantly higher risk for recurrences than patients who had recovered from a pure (non-mixed) manic state. Lithium and the combination of lithium and imipramine were highly effective preventive treatments for the pure manic group and poor treatments for the mixed group. Imipramine was ineffective for both the pure and mixed groups. The need for identifying mixed mania in therapeutic trials and in evaluating alternative treatments for lithium with this subgroup is discussed. PMID- 2970499 TI - Plasma dexamethasone and the dexamethasone suppression test. Initial and follow up tests in depressed patients. AB - Plasma dexamethasone concentrations following oral dexamethasone administration were examined in 78 patients with major depression prior to and during treatment. The test-retest stability of plasma dexamethasone levels within patients was satisfactory with an overall significant positive correlation between tests for each patient. However, significant variability was noted in individual patients. Change in pre-DST cortisol and plasma dexamethasone levels were the two variables, in that order of importance, contributing to change in DST status. In studies examining the clinical utility of serial dexamethasone suppression tests as a guide to recovery from depression, the effect of variability in plasma dexamethasone concentrations should be taken into account. PMID- 2970500 TI - Effects of naloxone on adrenal cortex regulation in patients with primary aldosteronism. AB - Excess production of proopiomelanocortin (POMC)-derived peptides with aldosterone stimulating activity has been suggested to play a pathogenetic role in idiopathic hyperaldosteronism (IHA). To further investigate this issue, the opiate receptor antagonist naloxone was administered to 14 patients with primary aldosteronism, 6 with an aldosterone-producing adenoma (APA) and 8 with IHA. Clinical and hormonal effects of iv administration of naloxone (10 mg as a bolus) were compared with those obtained in 8 normal subjects. In normals as well as in APA and IHA patients, naloxone caused a significant increase in plasma cortisol, and no change in ACTH, plasma renin activity (PRA) and aldosterone levels. All subjects were retested after 2 mg dexamethasone. ACTH and cortisol were reduced and PRA was unchanged in all groups, without modifications after naloxone. Baseline aldosterone showed no significant changes in all groups. While normal subjects and APA failed to show any aldosterone response to naloxone after dexamethasone, IHA patients demonstrated a significant decrease. beta-endorphin concentrations were in the normal range before and after dexamethasone. In conclusion, naloxone may have a direct action upon adrenal zona fasciculata increasing the cortisol responsiveness to physiological levels of ACTH in either normals or APA and IHA patients. The decrease of aldosterone induced by naloxone in IHA may be due to an intraadrenal opioid control of zona glomerulosa in this disorder. PMID- 2970501 TI - Persistence of specific antibodies after hepatitis B vaccination. AB - Antibody levels to hepatitis B surface antigen (anti-HBs) in healthy adults vaccinated with three doses of plasma-derived hepatitis B vaccine, containing 20, 10 or 5 micrograms of antigen, were followed for 4-6 years. After vaccination, 1034 of 1076 individuals had developed anti-HBs and 1016 had antibody concentrations above 10 IU/l. 681 of all initial responders could be tested after 1 year, 520 after 2, 380 after 3 and 213 after 4 years. 72 and 39 individuals, respectively, of the 185 earliest vaccinated volunteers were available for retesting after 5 and 6 years. Four years after the first vaccination, anti-HBs levels in 34% had dropped below 10 IU/l. The persistence of anti-HBs above this value depended on the peak antibody response after the third vaccination. Whereas all vaccinees tested with peak anti-HBs levels above 10,000 IU/l still had levels above 10 IU/l after 6 years, no-one with initial values between 10 and 100 IU/l maintained antibody concentrations above 10 IU/l for longer than 4 years. The rate of decrease in anti-HBs was independent of the peak anti-HBs value, the vaccine dose, and the age and sex of the vaccinees. PMID- 2970502 TI - Detection and quantification of anti-HBs antibodies by the Czechoslovak ELISA SEVAC Kit (Sevatest ELISA anti-HBs). AB - We invented and verified the possibilities of clinical use of the ELISA Kit for quantification of human serum anti-HBs antibodies. The kit does not employ any labelled antigen but is based on the principle of neutralization. The kit was tested on the panel of sera of the normal population, blood donors, workers at risk workplaces and persons vaccinated with Heptavax-B vaccine. The sensitivity of the kit is 80 mIU/ml, its capture is only slightly worse (in the convalescents) than the capture of the AUSAB (Abbott, USA). Statistical values and frequency distribution at various levels of anti-HBs antibodies in the followed-up group, have been indicated. The kit can be used very well for the follow-up of patients afflicted with virus hepatitis B, for the observation of workers at risk workplaces, for selection of persons eligible for vaccination and for checking the vaccination. It is suitable for the selection of sera for the preparation of specific globulin against hepatitis B and for standardization of this preparation. It is less suited for the compilation of epidemiological surveys where it is important to detect even as low levels of antibodies as 10-80 mIU/ml. PMID- 2970503 TI - The effect of withdrawing antihypertensive therapy: a review. AB - In this report the effects of withdrawing antihypertensive medication are reviewed from case reports and randomized trials. Success, defined as the proportion of patients remaining normotensive after withdrawal of medication varied from 15% to over 50%. Factors that predicted the return of high blood pressure after withdrawal, however, included high level of pre-treated blood pressure, marked obesity, short duration of treatment and left ventricular hypertrophy (LVH). Male patients also tended to be more likely to return to having high blood pressure than female patients. In community studies, it is predicted that many subjects will not be suitable candidates for withdrawal or treatment in view of their high blood pressures (both treated and pre-treated), obesity, and also their unwillingness to stop therapy. PMID- 2970504 TI - The functionally distinct subpopulations of human CD4+ helper/inducer T lymphocytes defined by anti-CD45R antibodies derive sequentially from a differentiation pathway that is regulated by activation-dependent post-thymic differentiation. AB - The CD4+ helper/inducer T cell population is comprised of functionally distinct subsets identifiable by the HB11 (anti-CD45R) mAb. We have previously shown that the cells that provide help for antibody production express the CD4+CD45R- phenotype. In contrast, CD4+ CD45R+ cells have minimal, if any, helper cell functions; rather, these cells function as inducers of Ts cell activity. The lineal relationship of these phenotypically and functionally distinct CD4+ subsets is unknown. In the present studies, we have examined the hypothesis that the CD4+ subpopulations identifiable with anti-CD45R antibodies represent "virgin" or "memory" T cells sequentially derived from a common differentiation pathway but differing in their relative maturation. When freshly purified cells were tested, CD4+ CD45R+ cells had no Th cell function. However, after in vitro activation with PHA and propagation in IL-2, CD4+CD45R+ cells acquired the ability to provide help for antibody production. Moreover, this functional acquisition by these cells was accompanied by their conversion to the CD4+CD45R- phenotype. Analyses of the activation, growth kinetics, and functional dose response characteristics of CD4+CD45R+ and CD4+CD45R- cells demonstrated that our findings did not result from the selective growth of CD4+ CD45R- cells contaminating the CD4+CD45R+ preparations. Thus, these data demonstrate that the "helper" and "suppressor-inducer" subsets of CD4+ cells identified by anti-CD45R antibodies are not comprised of fully mature, phenotypically and functionally stable T cells. Rather, these CD4+ subsets appear to represent cells at different maturational stages of an activation-dependent, post-thymic differentiation pathway. PMID- 2970505 TI - Localization of the receptor binding site of IFN-alpha 2b. AB - The receptor binding site of IFN-alpha is not precisely known. To further characterize this site, mAb against IFN-alpha 2b were selected that block the binding of radiolabeled IFN-alpha 2b to its cell surface receptor. These antibodies also neutralized the anti-viral and anti-proliferative properties of IFN-alpha 2b. A subset of these antibodies (group 1) do not recognize IFN-alpha 2a, either in solid-phase immunoassays or functional assays, whereas a second subset (group 2), with no cross-reactivity with group 1, recognizes both IFN alpha subtypes. Because IFN-alpha 2b and IFN-alpha 2a differ by only alpha Arg23 Lys23 substitution, group 1 antibodies must recognize an epitope within the receptor binding region of IFN-alpha 2b that includes Arg23. Group 2 antibodies recognize a separate and distinct epitope within the binding site that does not include Arg23. PMID- 2970506 TI - IL-1 as a co-factor for lymphokine-secreting CD8+ murine T cells. AB - Immunologically important among the known biologic activities of IL-1 is its ability to function as a co-factor for responses mediated by lymphokine secreting CD4+ Th cells. In contrast to its known effects in CD4+ T cell responses, IL-1 is not known to play a role in CD8+ T cell responses. In the present study, we have assessed the ability of murine recombinant IL-1 to function as a co-factor for stimulating CD8+ T cells to secrete lymphokines such as IL-2. We found that, in conjunction with either Ag or mitogen, IL-1 is able to stimulate lymphokine secreting CD8+ T cells. Furthermore, we found that, as a consequence of its stimulation of lymphokine-secreting CD8+ T cells, IL-1 is able to reconstitute MHC class I allospecific cytolytic T lymphocyte responses by cell populations depleted of both accessory cells and CD4+ T cells. These results demonstrate that the biologic activity of IL-1 is not restricted to CD4+ cell responses, and suggests that IL-1 can function as a co-factor for the stimulation of lymphokine secreting Th cells regardless of their CD4/CD8 phenotype. If IL1 acts directly on lymphokine-secreting T cells or on the APC with which they interact is not yet certain. PMID- 2970507 TI - The characterization of four monoclonal antibodies specific for mouse IL-5 and development of mouse and human IL-5 enzyme-linked immunosorbent. AB - Four rat mAb directed against mouse IL-5 have been characterized by their ability to remove and neutralize mouse IL-5 activity in various bioassays. All four mAb absorbed IL-5 activity from solution. Although all were able to neutralize mouse IL-5 bioactivity, two were significantly more effective. These two were also able to neutralize the activity of mouse IL-5 delivered to B cells during a cognate linked interaction with a Th cell clone. A two-site sandwich ELISA specific for mouse IL-5 was developed by using pairs of mAb. The mouse IL-5 ELISA is capable of detecting natural or mouse rIL-5 in supernatants, crude bacterial lysates, and high concentrations of mouse serum, and has a detection limit of less than 20 pg. Two of these antibodies cross-reacted with and neutralized human rIL-5, and one of these was used for development of an ELISA for human IL-5. PMID- 2970508 TI - Regulation of complement factor H synthesis in U-937 cells by phorbol myristate acetate, lipopolysaccharide, and IL-1. AB - The capacity of the human monocyte cell line U-937 to synthesize complement factor H was examined. The kinetics of secretion of factor H into cell culture supernatant were followed by a sensitive solid phase RIA capable of measuring 0.1 ng of protein. Daily secretion of factor H was low and almost linear and was approximately 3 ng of factor H per 10(6) cells. Factor H synthesis was inhibited by cycloheximide but returned to the levels seen in untreated cultures after removal of the inhibitor. LPS and IL-1 both effected a time- and dose-dependent enhancement of factor H synthesis. Induction of U-937 cells with PMA to differentiate into macrophage-like cells also resulted in increased factor H synthesis. RIA of cell lysates, immunofluorescence microscopy, as well as FACS analysis all revealed that factor H Ag was also associated with the U-937 cell membrane. The population of U-937 cells bearing membrane-associated factor H was decreased from 77 to 43% after stimulation for 48 h with LPS (1 microgram/ml). [35S]Methionine metabolic labeling and SDS-PAGE analysis of factor H immunoprecipitates from unstimulated and stimulated culture supernatants and cell lysates demonstrated a major polypeptide, m.w. 150,000, and a minor component, m.w. 42,000. Western blot analysis of factor H in fresh normal plasma also detected both 150,000 and 42,000 m.w. factor H proteins. This is in agreement with the recent demonstration of a 4.4- and 1.8-kb mRNA for factor H in human liver. These data demonstrate that U-937 cells synthesize factor H that is structurally and antigenically similar to factor H in normal plasma. The exact nature of the membrane-bound factor H and its functions and mechanism of attachment to the cell membrane remain to be elucidated. PMID- 2970509 TI - Genetic control of murine T cell proliferative responses to Mycobacterium leprae. V. Evidence for cross-reactivity between host antigens and Mycobacterium leprae. AB - T cell proliferative responses to Mycobacterium leprae were measured by immunization of mice at the base of the tail with Ag and challenging lymphocytes from draining lymph nodes in culture with M. leprae. C57BL/10J and B10.BR mice were identified as low responder mice and the congenic strains B10.M, B10.Q, and B10.AKM as high responders whereas F1 (high x low) hybrid mice were found to be low responders. The cellular basis of low responsiveness did not appear to result from a defect in Ag-presenting cells or the activation of suppressor T cells by M. leprae. The influence of the environment in which T cells developed on responsiveness to M. leprae was analyzed in chimeric mice prepared by irradiating F1(C57BL/10J x B10.M) mice and reconstituting with bone marrow from C57BL/10J, B10.M, or F1 donors. Six weeks later, chimeric mice were immunized with M. leprae, lymph node cells were subsequently prepared, and H-2 phenotyped and challenged in culture with M. leprae Ag. T cell proliferative responses were found to be low in all cases, similar to those observed using lymph node cells from F1 hybrid mice. These results suggested that high responder B10.M lymphocytes developing in the irradiated F1 mice became tolerized to antigenic determinants found on M. leprae. This implied cross-reactive epitopes existed between some mouse strains and M. leprae. Low responsiveness to M. leprae in low responder and F1 hybrid mice may result from tolerance to H-2-encoded Ag that show cross-reactivity with M. leprae. PMID- 2970511 TI - Position paper on short term strategies to resolve the nursing shortage May 5, 1988. PMID- 2970510 TI - Functional characterization of murine cell lines expressing high, intermediate, or negative levels of surface receptors for interferon-gamma. AB - Using radioiodinated murine interferon-gamma (IFN-gamma), various cell lines were analyzed for their capacity to bind IFN-gamma and to respond to its biological activity as measured by activation of the 2',5'-oligoadenylate (2-5A) synthetase, induction of the antiviral activity, and inhibition of [3H]thymidine uptake. A T cell lymphoma, EL-4, was found to express relatively high number of specific membrane receptors for IFN-gamma (60,000/cell) with a dissociation constant of around 9 X 10(-10) M. By contrast, a fibroblast cell line, K-BALB, transformed by the Kirstein murine sarcoma virus, was found negative for binding of radioiodinated IFN-gamma, even in the presence of high ligand concentrations. Other cell lines displayed numbers of receptors varying between 650 to 20,000 binding sites/cell with KD ranging between 5 X 10(-9) to 9 X 10(-10) M. When incubated with EL-4 expressing high levels of specific receptors, IFN-gamma caused stimulation of cell growth, but not resistance to viral (vesicular stomatitis virus) infection and expression of 2-5A synthetase. By contrast, when L1210 and TS/A cells, expressing intermediate levels of specific receptors, were incubated with IFN-gamma, inhibition of cell growth, induction of antiviral resistance and activation of 2-5A synthetase was observed. Finally, K-BALB cells, lacking specific receptors for IFN-gamma, were completely unsensitive to its biological activity. The present system could represent a useful model for the characterization of the interaction of IFN-gamma with target cells at molecular levels. PMID- 2970512 TI - Beta-endorphin levels in the preovulatory follicles and the outcome of in vitro fertilization. AB - The level of immunoreactive beta-endorphin-like (IR-beta-EP) material(s) was determined in follicular fluid samples obtained from patients treated for in vitro fertilization (IVF). Follicles containing morphologically mature oocytes had slightly lower concentrations of IR-beta-EP. The total IR-beta-EP content was similar in follicles with oocytes at different degrees of maturity. A smaller amount of IR-beta-EP in the follicle fluid was associated with a greater developmental potential of the oocytes, which formed embryos that reached the three- to six-cell stage at 40-44 hr after insemination (r = -0.35, P less than 0.026). A higher IR-beta-EP content was seen in fluid samples containing oocytes that remained uncleaved after fertilization (P less than 0.05 by Duncan's multiple-range test). PMID- 2970513 TI - Changes in small intestinal epithelial expression of MHC class II antigen after terminal ileal resection for Crohn's disease. AB - Aphthous lesions in the neoterminal ileum from patients operated for Crohn's disease are an early sign of recurrence that can be identified during ileocolonoscopy. The origin of these lesions was studied in nine patients treated by terminal ileal resection and right hemicolectomy for complicated Crohn's disease. During surgery the neoterminal ileum was turned inside out, the mucosa was carefully inspected and two large mucosal biopsies were obtained. The same procedure was carried out in seven patients operated for other diseases. Four to six months after surgery endoscopy of the neoterminal ileum was carried out and multiple biopsies were obtained from the neoterminal ileum. Another follow-up colonoscopy with biopsies was carried out one year after the operation. The operative specimens and the per- and postoperative biopsies were submitted to routine microscopy and immuno- and enzyme-histochemistry. None of the Crohn's patients had macroscopic lesions in the neoterminal ileum at operation and only one had microscopic signs of inflammation and a positive section margin. Four-six months after operation all Crohn's patients had active aphthous lesions in a 5-20 cm segment of the neoterminal ileum at endoscopy. Biopsies taken at this time showed microscopic features which were not observed in biopsies from control subjects: an increase of HLA-DR+, ATPase+ dendritic cells in the ileal mucosa and a defective expression of MHC class II antigens by the small intestinal epithelial cells. MHC class II expression by the small intestinal epithelial cells returned towards normal after one year.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970514 TI - Contact sensitivity to gentamicin-hydrocortisone ear drops. AB - Patch testing of ten patients who had been treated with gentamicin-hydrocortisone ear drops for active chronic suppurative otitis media showed evidence of contact sensitivity in five. Positive skin tests correlated with failure to respond to treatment. PMID- 2970515 TI - Up-regulation of gamma interferon receptors on the human monocytic cell line U937 by 1,25-dihydroxyvitamin D3 and granulocyte-macrophage colony stimulating factor. AB - The human monoblast-like cell line U937 can be induced to differentiate by a variety of agents including phorbol esters, retinoic acid, gamma interferon (IFN gamma), and 1,25-dihydroxyvitamin D3 (VD3). Increased expression of OKM1 antigen, Fc receptors, and other cell surface antigens occur with the differentiation of this cell line along the macrophage lineage. Whereas 10(-8) M VD3 alone induces changes in cell surface antigens, there were no changes in the number or affinity of IFN gamma receptors. Incubation of U937 with VD3 and 100 U/ml of granulocyte macrophage colony-stimulating factor (GM-CSF) resulted in further increases in OKM1 antigen expression and an up-regulation of IFN gamma receptors. The number of IFN gamma receptors increased between two- and fourfold and was maximal after 48 h incubation with VD3 and GM-CSF. Scatchard analysis revealed a single class of receptors before or after differentiation, although the increase in receptor number was associated with an overall decrease in receptor-binding affinity. Incubation of U937 with VD3 plus GM-CSF and IFN gamma resulted in further increases in the density of OKM1 antigen expressed per cell. This increase in OKM1 expression was greater than that observed for U937 incubated with VD3 and GM CSF or VD3 and IFN gamma alone. These results suggest that GM-CSF up-regulates IFN gamma receptors on VD3-stimulated U937 and enables these cells to be induced further along the pathway of macrophage differentiation, possibly by subsequent interaction with additional cytokines such as IFN gamma. PMID- 2970516 TI - Mechanisms responsible for mediating the antidiuretic action of vasopressin. PMID- 2970517 TI - Two subsets of human T lymphocytes expressing gamma/delta antigen receptor are identifiable by monoclonal antibodies directed to two distinct molecular forms of the receptor. AB - Two mAbs directed to the TCR-gamma/delta were analyzed for their pattern of reactivity with CD3+WT31- cell populations or clones. In normal individuals, the BB3 mAb reacted with approximately 2/3 of peripheral blood CD3+WT31- lymphocytes, whereas delta-TCS-1 stained approximately 1/3 of such cells. In addition, the sum of the percentages of BB3+ and delta-TCS-1+ cells approximated the percentages of peripheral blood CD3+WT31- lymphocytes in seven normal donors tested. Also, in peripheral blood-derived polyclonal CD3+WT31- populations, cultured in IL-2, cells reacting with one or another mAb accounted for the whole cell population. On the other hand, only delta-TCS-1-reactive cells, but not BB3+ cells, could be detected in unfractionated as well as in CD4-8-thymocyte populations. Analysis of peripheral blood-derived CD3+WT31- clones showed that 70% of 72 clones analyzed reacted with BB3 mAb, but not with delta-TCS-1 mAb. On the other hand, delta-TCS 1 mAb stained the remaining BB3- clones. Five clones expressing medium-low amounts of CD8 antigen were BB3- delta-TCS-1+. Both types of clones lysed the Fc gamma receptor-bearing P815 target cell in the presence of anti-CD3 mAb (but not of mAb directed against HLA-DR, CD7 molecules, or TCR-alpha/beta). In this cytolytic assay, BB3 mAb induced target cell lysis only by BB3+ clones, whereas delta-TCS-1 mAb was effective only with delta-TCS-1+ clones. The CD3-associated surface molecules expressed by BB3+ or delta-TCS-1+ clones were analyzed after cell surface iodination and immunoprecipitation with the corresponding anti-TCR mAb or with anti-CD3 mAb (in digitonin-containing buffer). In SDS-PAGE, molecules immunoprecipitated from 13 BB3+ clones displayed, under nonreducing conditions, a molecular weight of 80 kD (in some cases, a minor 38-kD band could be detected). Under reducing conditions, two major components of 44 and 41 kD (and a minor component of 38 kD) were detected. On the other hand, TCR molecules immunoprecipitated from 11 different delta-TCS-1+ clones appeared as a diffuse band of 41-44 kD, both under reducing and nonreducing conditions (under non reducing condition, an additional 38-kD band was present). Therefore, BB3+ cells express a disulphide-linked form of TCR-gamma/delta whereas delta-TCS-1+ cells express a non-disulphide-linked form.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970518 TI - Lymphokine-mediated regulation of the proliferative response of clones of T helper 1 and T helper 2 cells. AB - Murine Th1 and Th2 subsets differ not only in the lymphokines they produce, but also functionally. It is not clear what factors influence the preferential activation of one subset versus the other and what regulatory interactions exist between them. The purpose of this study was to examine the effect of lymphokines produced by clones of Th1 cells (IL-2 and IFN-gamma), Th2 cells (IL-4), and APC (IL-1) on the proliferative response of Th1 and Th2 cells after antigenic stimulation. Activation of both types of clones in the presence of antigen and APC resulted in the acquisition of responsiveness to the proliferative effects of both IL-2 and IL-4, although Th2 cells were more responsive to IL-4 than Th1 cells. Responsiveness of Th1 and Th2 cells to both lymphokines decreased with time after initial antigenic activation; Th1 cells lost their responsiveness to IL-4 more rapidly and to IL-2 more slowly than Th2 cells. IFN-gamma partially inhibited the IL-2 and IL-4-mediated proliferation of Th2, but not Th1 cells. Although the presence of IL-1 was not required for the response of Th1 or Th2 cells to IL-4, its presence resulted in a synergistic effect with IL-2 or IL-4 in Th2 but not in Th1 cells. Both subsets responded to a mixture of IL-2 and IL-4 in synergistic fashion. Delayed addition and wash-out experiments indicated that both IL-2 and IL-4 had to be present simultaneously in order for synergy to occur. These results suggest that Th cell subsets might regulate each other via the lymphokines that they secrete and that the pathways of IL-2 and IL-4 mediated proliferation are interrelated. PMID- 2970519 TI - IgE-antigen complexes enhance Fc epsilon R and Ia expression by murine B lymphocytes. AB - Murine monoclonal IgE interacts with B cells of BALB/c mouse spleen with greater efficiency in the presence of its specific antigen. Complexes of anti-DNP IgE and DNP-OVA not only resist elution from B lymphocytes by acid but have a substantially longer dissociation half-time when compared with monomeric IgE (440 vs. 8 min, respectively). Further, these IgE-antigen complexes induce Fc epsilon R expression in lymphoid cells more efficiently than IgE alone. Maximum levels of B cell Fc epsilon R were observed after a 24 h incubation with 1 microgram/ml IgE in the presence of 1 microgram/ml DNP-OVA, while 30 micrograms/ml monomeric IgE was needed to elicit a similar increase of Fc epsilon R expression. Most importantly, overnight incubation of B cell-enriched BALB/c spleen cells with IgE antigen complexes resulted in an augmented membrane expression of class II MHC antigens. B cell surface expression of both I-A and I-E antigens responded to a comparable level after incubation with IgE-antigen complexes but did not occur in response to either IgE or antigen alone. The enhanced sIa expression occurred in parallel to IgE-antigen concentrations that gave rise to Fc epsilon R hyperexpression. Moreover, double staining for Fc epsilon R and surface Ia antigen shows that B cells exhibiting the highest density of Fc epsilon R also demonstrated the most surface I-A, suggesting that B lymphocytes are autonomous in their response to IgE-antigen complexes. Changes in class I MHC or sIgM were not observed after overnight incubation with IgE and antigen. These results demonstrate the importance of IgE-antigen complexes for intercellular signaling and further suggest that the IgE system plays a broader role in immune response than it has generally been credited. PMID- 2970520 TI - In vivo stimulation and restoration of the immune response by the noninflammatory fragment 163-171 of human interleukin 1 beta. AB - The synthetic nonapeptide VQGEESNDK, corresponding to the fragment 163-171 of human IL-1 beta, showed in vivo immunomodulatory capacities qualitatively and quantitatively comparable to those of the mature human IL-1 beta protein. In fact, both IL-1 beta and the 163-171 fragment stimulated the immune response of normal mice and restored immune reactivities of immunocompromised animals. In addition, the synthetic IL-1 peptide was as efficient as the entire protein in inducing tumor rejection and radioprotection. On the other hand, the 163-171 fragment did not cause any of several inflammation-associated metabolic changes inducible by the whole IL-1 beta molecule in vivo: hypoferremia, hypoglycemia, hyperinsulinemia, increase in circulating corticosterone, SAA and fibrinogen, decrease in hepatic drug-metabolizing enzymes. Furthermore, at variance with IL-1 beta, the 163-171 peptide did not show the toxic effects causing shock and death in adrenalectomized mice. Thus, these results confirm our previous in vitro observations that functional domains are identifiable within the multipotent cytokine IL-1 beta, and demonstrate the biological relevance of this finding in a variety of in vivo systems. The identification of a selectively active fragment of a cytokine may thus represent a significant step towards a better directed and more rational immunotherapeutic approach. PMID- 2970521 TI - Lymphokine production by murine T cells in the mixed leukocyte reaction. AB - Although the production of B cell stimulatory factors by cell lines and hybridomas is well established, production of specific lymphokines by normal T cells in response to antigen stimulation has not been analyzed. We have used bioassays and neutralizing mAbs to demonstrate that IL-2, IL-4, and B cell growth factors (BCGF) are produced during primary and secondary MLRs. IL-2 is detected in the first 12 h of both types of MLR. IL-4 and BCGF appear at 24-48 h in the conditioned medium of the primary MLR, and peak by 12 h in the secondary MLR. The amount of IL-4 in the primary response reaches a level that is 10% of that detected in the secondary. In contrast, BCGF production steadily increases over time in the primary MLR, and maximal production is equivalent to that made in the secondary response. Allogeneic dendritic cells and anti-Ig-activated B blasts both stimulated lymphokine production in the primary MLR, whereas small B cells were weak. In the secondary MLR, all three cell populations stimulated the production of IL-2, IL-4, and BCGF. Therefore, the release of several defined B cell stimulating factors can be detected in the conditioned media of responding primary T lymphocytes. PMID- 2970522 TI - Inactivation of a T cell receptor-associated GTP-binding protein by antibody induced modulation of the T cell receptor/CD3 complex. AB - TCR modulation induced by anti-TCR or anti-CD3 mAbs leads to a transient state of refractoriness of the T cell to all signals given via cell surface structures. To investigate the underlying mechanisms, we have used human CTL permeabilized with the alpha toxin of S. aureus. This method of permeabilization allows manipulation of the interior milieu of the cell, but maintains its functional and structural integrity. Introduction of the G protein activator GTP gamma S into permeabilized CTL leads to triggering of granule exocytosis. The G protein inactivator GDP beta S inhibited exocytosis induced by TCR triggering but not that induced by activation of protein kinase C. This indicates that the G protein that triggers exocytosis is localized after CD3 triggering but before formation of the polyphosphoinositol breakdown product diacylglycerol. In TCR-modulated CTL, GTP gamma S is no longer able to activate exocytosis. Such CTL, however, still respond to PKC activators. This demonstrates that a TCR-associated G protein has been functionally inactivated by TCR modulation. PMID- 2970523 TI - Semliki Forest virus-specific non-structural protein nsP3 is a phosphoprotein. AB - Antisera were raised in rabbits against fusion proteins consisting of beta galactosidase and partial amino acid sequences of Semliki Forest virus (SFV) specific non-structural proteins nsP1, nsP2, nsP3 and nsP4. The antisera were specific since each of them precipitated only one labelled protein of a size expected for nsP1, nsP2, nsP3 or nsP4 from lysates of [35S]methionine-labelled SFV-infected BHK-21 cells. The specific antisera also precipitated p220 (with sequences of nsP1, nsP2 and nsP3), p155 (nsP1 and nsP2) and p135 (nsP3 and nsP4) which have been previously shown to be cleavage products of the polyprotein precursor of the non-structural proteins. nsP1, nsP4 and most of nsP3, together with the virus-specific RNA polymerase activity, were in the mitochondrial pellet (P15) fraction of infected BHK-21 cells whereas nsP2 was evenly distributed between P15 and the supernatant fraction (S15). Only antisera directed against nsP3 sequences precipitated a labelled protein from cells incubated with [32P]orthophosphate during SFV infection. Treatment of the immunoprecipitate with calf alkaline intestinal phosphatase reduced the amount of labelled nsP3 considerably. Immunoprecipitated 32P-labelled nsP3, isolated by SDS-PAGE, was subjected to acid hydrolysis. Both phosphoserine and phosphothreonine but not phosphotyrosine could be identified in the hydrolysate. Approximately twice as much [32P]serine as [32P]threonine was detected in nsP3. P15 and S15 fractions were prepared from [35S]methionine- and 32P-labelled SFV-infected cells and the 35S/32P ratio of nsP3 was determined after immunoprecipitation and SDS-PAGE. The nsP3 in S15 was less heavily phosphorylated (about 50%) than P15-associated nsP3. Anti-nsP3 serum revealed large cytoplasmic vesicles in SFV-infected cells in indirect immunofluorescence microscopy. PMID- 2970524 TI - Brain indoleamines in alloxan- and streptozotocin-induced diabetic rats. AB - Previous work by other authors has shown that alloxan-induced diabetes increases whereas streptozotocin-induced diabetes does not alter nonesterified fatty acid (NEFA) plasma levels. The present study replicates these results and demonstrates that fasted, streptozotocin-induced diabetic animals also have increased NEFA levels. In addition, brain levels of 5-hydroxytryptamine (5-HT) and of its immediate precursor and metabolite were measured. Alloxan- and fasted, streptozotocin-induced diabetic rats showed significant increases in brain indoleamine concentrations, whereas fed, streptozotocin-induced diabetic rats had unchanged levels of the same compounds. Levels of brain indoleamines exhibited a strong positive correlation with wet-dog shakes (an index of 5-HT activity) elicited by hippocampal stimulation. Blockade of wet-dog shakes by 5-HT receptor antagonists strengthens the proposal that this behavior is a good index of central 5-HT activity. The increased content of brain indoleamines in alloxan- and fasted, streptozotocin-induced diabetic rats may be related to the increased NEFA plasma levels seen in the same animals. This hypothesis is supported by the positive correlation demonstrated between NEFA and 5-HT levels. In conclusion, it is suggested that alloxan-induced diabetes may represent a useful model for studying the various behavioral changes known to occur in diabetics. PMID- 2970525 TI - Modulation by docosahexaenoic acid of the epinephrine-stimulated adenylate cyclase activity of the bovine retina. AB - This work shows that unsaturated fatty acids enhance the epinephrine-stimulated adenylate cyclase activity in bovine retina. The modulating effect on the epinephrine-stimulated formation of cyclic AMP seems to be linked to the degree of unsaturation of the fatty acid. Treatment of the intact retina with docosahexaenoic acid in the concentration range 0.5 X 10(-6)-1 X 10(-3) M does not affect the enzyme activity measured in the absence of the hormone but markedly increases the cyclase activity when the tissue is incubated in the presence of 0.1 mM epinephrine. Docosahexaenoic acid enhances the maximal response to epinephrine without affecting the apparent ED50 value for this effector. Docosahexaenoic acid at 0.5 mM also increases the hormone-stimulated adenylate cyclase activity in retinal cell-free homogenate, whereas it has no effect on the epinephrine-sensitive enzyme solubilized from the membrane fraction with 1% Triton X-305. When docosahexaenoic acid-preincubated intact retina and cell-free homogenate are incubated in the presence of defatted albumin, both the observed activating effect of the fatty acid on the epinephrine-stimulated adenylate cyclase activity and the enhancement of the enzyme response to the hormone significantly diminish. PMID- 2970527 TI - D1 and D2 dopamine receptors in caudate-putamen of nonhuman primates (Macaca fascicularis). AB - D1 and D2 dopamine receptors were characterized in the caudate-putamen region of nonhuman primate brains (Macaca fascicularis). D1 dopamine receptors were identified with [3H]SCH 23390 and D2 receptors with [3H]-spiperone. Scatchard analysis of [3H]SCH 23390 saturation data using washed membranes revealed a single high-affinity binding site (KD, 0.352 +/- 0.027 nM) with a density (Bmax) of 35.7 +/- 2.68 pmol/g original wet tissue weight (n = 10). The affinity of [3H]spiperone for the D2 site was 0.039 +/- 0.007 nM and the density was 25.7 +/- 1.97 pmol/g original wet tissue weight (n = 10). D1 and D2 receptors in nonhuman primates may be differentiated on the basis of drug affinities and stereoselectivity. In competition experiments, RS-SKF 38393 was the most selective D1 agonist, whereas (+)-4-propyl-9-hydroxynaphthoxazine [(+)-PHNO] was the most selective D2 agonist. Apomorphine was essentially nonselective for D1 or D2 binding sites. Of the antagonists, R-SKF 83566 and SCH 23390 were the most selective for the D1 site, whereas YM-09151-2 was the most selective for the D2 site. cis-Flupentixol and (S)-butaclamol were the least selective dopamine antagonists. D1 receptors bound benzazepine antagonists (SCH 23390/SCH 23388, R SKF 83692/RS-SKF 83692) stereoselectively whereas D2 receptors did not. Conversely D2 receptors bound (S)-sulpiride and (+)-PHNO more potently than their enantiomers whereas D1 receptors showed little stereoselectively for each of these isomeric pairs. These binding characteristics may be utilized for evaluation of individual receptor function in vivo. PMID- 2970526 TI - Studies on the metabolism and toxicity of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine in cultures of embryonic rat mesencephalon. AB - The metabolism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was studied in dissociated cell cultures prepared from embryonic rat mesencephalon. MPTP was added to the feeding medium and at the end of the incubation period MPTP was separated from the water-soluble metabolite by ether extraction. Our results demonstrate that, in cultures, MPTP is oxidized predominantly by monoamine oxidase B, since deprenyl but not clorgyline had an inhibitory effect on its metabolism. The metabolite of MPTP diffuses freely in the feeding medium (99% of the total) and its concentration increases with time. The concentration of the metabolite can be increased by increasing the number of cells plated into the tissue culture dish and the toxicity to dopamine neurons depends on the amount of metabolite accumulated in the feeding medium. The metabolism of MPTP is reduced by decreasing the number of glial cells present in the cultures. PMID- 2970529 TI - Various features of various disability contracts. PMID- 2970528 TI - A point mutation in the coding sequence of the beta-hexosaminidase alpha gene results in defective processing of the enzyme protein in an unusual GM2 gangliosidosis variant. AB - cDNA clones were isolated from cultured fibroblasts of a patient previously reported as having GM2-gangliosidosis due to defective processing of the precursor beta-hexosaminidase alpha chain. Sequence analysis of a clone containing the entire protein coding sequence showed a single nucleotide substitution, from G to A, at nucleotide residue no. 1444, which resulted in a change in amino acid residue no. 482, from the normal glutamic acid to lysine. This transversion was confirmed in two other cDNAs from the same unamplified library. The results collectively indicate that the change from the strongly negative to strongly positive charge at amino acid residue no. 482 is responsible for the defective processing of the enzyme in this patient. PMID- 2970530 TI - Risks and potential legal problems associated with treatment of human immunodeficiency virus (HIV) positive patients. PMID- 2970531 TI - Heparan sulfate proteoglycan and laminin immunoreactivity on cultured astrocytes: relationship to differentiation and neurite growth. AB - Extracellular matrix (ECM) produced by Schwann cells is known to promote growth of several types of neurites (Ard et al., 1987). Whether a similar material produced by astrocytes may be available to promote neurite growth during CNS development is now open to question. The present study was undertaken to define conditions under which cultured astrocytes deposit the ECM components laminin and heparan sulfate proteoglycan (HSPG), and to relate this deposition to the ability of astrocytes to support neurite growth. The use of 2 different culture media permitted the growth of astrocytes either with or without these ECM components. Neonatal rat cortical astrocytes were cultured by the method of McCarthy and de Vellis (1980) and studied by immunocytochemistry and electron microscopy. Astrocytes grown in serum-containing medium for 5 or 9 d after subculturing were shown to have fibrillar patches of ECM containing both HSPG and laminin immunoreactivity. Immunoreactivity for the 2 molecules was usually colocalized. In contrast, astrocytes subcultured for 5 d in defined medium showed no immunocytochemical staining for either laminin or HSPG and had no ECM visible in EM. Formation of stellate processes was increased when cells were grown in defined medium compared with that seen in serum-containing medium, and growth of the population was slower. In 3 other conditions, attainment of stellate morphological differentiation by the astrocytes was correlated with diminution in immunostaining for ECM components. (1) In older cultures (30-42 d after subculturing), stellate, mitotically quiescent cells showed relatively little HSPG or laminin immunoreactivity. (2) Cultures initially maintained in serum containing medium and then converted to defined medium lost much of their immunoreactivity for ECM components and developed longer processes. (3) When neurites from fetal rat dorsal root ganglion explants grew across monolayers of astrocytes in serum-containing medium, diminution of ECM immunostaining and development of stellate processes were seen in areas directly contacted by the neurites. ECM-containing laminin and HSPG did not appear to be necessary for neurites to interact with astrocytes. In defined medium, in which no ECM was detected, dorsal root ganglion neurites were found in contact with astrocyte surfaces rather than on the rat tail collagen substratum on the culture dish.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970532 TI - Technetium-99m methoxyisobutyl isonitrile (RP30) for quantification of myocardial ischemia and reperfusion in dogs. AB - This study was done to determine whether [99mTc]methoxyisobutyl isonitrile (RP30), a nonredistributing myocardial perfusion agent, could be used to quantify regional myocardial blood flow distribution during ischemia and reperfusion, employing sequential injections of tracer, tomographic imaging, and appropriate image subtraction. Dogs underwent transient (6 min) coronary artery occlusion, followed by two paired injections of RP30 and radioactive microspheres combined with tomographic imaging, the first during coronary occlusion and the second after 60 min of reperfusion. To obtain a true image representative of reperfusion, the first set of images was corrected for 99mTc decay and subtracted from the second. During occlusion, tissue microsphere content and scintigraphic RP30 activity in the center of the ischemic region (both expressed as a fraction of the nonischemic region) were closely correlated, although RP30 consistently exceeded microsphere content (0.43 +/- 0.03 vs. 0.24 +/- 0.04, p less than 0.01). Direct tissue counting of RP30 confirmed its relative excess in ischemic myocardium. Reperfusion was successful in 7/8 dogs, with an increase in RP30 activity to 0.98 +/- 0.04 compared to 0.89 +/- 0.03 for microspheres (p = N.S.). In one dog with microsphere-documented persistent ischemia, the RP30 defect was still present after reflow. Our results indicate that because of the lack of myocardial clearance and redistribution, repeat injections of RP30 can be used to quantify serial changes in regional myocardial blood flow. PMID- 2970533 TI - Treatment of patients with AIDS: a matter of professional ethics. PMID- 2970534 TI - Atrial natriuretic factor in the human fetus: effect of volume expansion. PMID- 2970535 TI - Mesaconic aciduria in patients treated with sodium benzoate. PMID- 2970536 TI - Characterization of suppressor T-cells induced with concanavalin A in vivo by cell electrophoresis. AB - The splenic T-cells of concanavalin A (Con-A)-injected mice suppressed the in vitro Con A-induced splenic blastogenesis of normal mice. We examined electrophoretic mobilities (EPM) and surface markers of Con A-induced suppressor T-cells as a model of suppressor T-cells. The EPM of the suppressor cells with both Thy-1 and gangliotetraosyl-ceramide (asialo GM1) was lower than that of normal T-cells. The ratio of low mobility T-cells (LMT) to high mobility T-cells (HMT) correlated significantly with dose-response and time-course of Con A in the suppressor activity and with the percentages of cells carrying asialo GM1 antigen and receptor for peanut agglutinin. For in vitro induction by Con A, Lyt-2+ cells were reported to have suppressor activity. However, for in vivo induction by Con A, Lyt-2+ cells did not correlate with the suppressor activity in the dose response and time-course experiments. By separating the Lyt-subsets, it was found that both Lyt-1+2+ and Lyt-1+2- subsets were suppressor cells in the blastogenesis. The suppressor activity of in vivo Con A-induced spleen cells correlated with the LMT/HMT ratio, but not with the analysis of Lyt-phenotype. These results suggested that lymphocytes electrophoresis clearly distinguished Con A-induced suppressor T-cells with low EPM from normal T-cells. PMID- 2970537 TI - Determination of 5-fluorouracil in plasma and liver after oral administration of 5'-deoxy-5-fluorouridine using gas chromatography-mass spectrometry. AB - A method for determination of 5-fluorouracil (5-FU) has been established for pharmacokinetic study of 5'-deoxy-5-fluorouridine (5'-DFUR), a newly developed prodrug of 5-FU. 5-FU was extracted with diethyl-ether containing 1-propanol and methylated with CH3I in the presence of (CH3)4NOH. The methylated 5-FU was analyzed by gas chromatography-mass spectrometry using 15N2-5-FU as an internal standard. Quantitation was carried out by selected-ion monitoring of molecular ions of N,N-dimethyl-5-FU (m/z 158) and the internal standard (m/z 160). The sensitivity (greater than 2 ng/g sample), specificity and precision of the method were satisfactory for application to clinical studies of this drug. After an oral administration of 5'-DFUR (800 mg/body) to patients with cancer, 5-FU in plasma peaked within 1 h and eliminated with an apparent half life of about 1 h. PMID- 2970538 TI - Increased tubuloglomerular feed-back mediated suppression of glomerular filtration during acute volume expansion in rats. AB - 1. Volume expansion is currently believed to change the intrinsic properties of the juxtaglomerular apparatus such that the sensitivity of the tubuloglomerular feedback (TGF) mechanism is reduced, thus allowing glomerular filtration rate, and hence salt and water excretion, to rise. Recent studies conflict with this view and indeed the older literature reveals that the rise in glomerular filtration rate (GFR) under these conditions is far more modest than would be expected if TGF control were eliminated. 2. To investigate this problem, TGF control of filtration rate was examined by measuring single-nephron glomerular filtration rate (SNGFR) during loop of Henle perfusion at varying rates in rats under control conditions, after acute, moderate (4% of body weight), iso-oncotic volume expansion and in rats treated with antibodies to atrial natriuretic peptide (ANP) prior to the acute volume expansion. 3. With TGF control of filtration interrupted by filtrate collection from the proximal tubule, SNGFR in the expanded rats was massively increased compared with controls, although SNGFR measured in the distal tubule, and hence with TGF control intact, was only modestly increased, as was whole-kidney filtration rate. Loop perfusion at increasing rates up to 30 nl min-1 progressively decreased SNGFR in controls, and in the expanded rats the range over which control was exerted extended up to 60 80 nl min-1. For changes in loop flow around the spontaneous operating point, the sensitivity of the TGF mechanism, defined as delta SNGFR/delta loop flow, was similar in both groups. Treatment of rats with ANP antibodies prior to volume expansion substantially blunted the changes in renal salt and water excretion and the increase in SNGFR seen in the absence of loop perfusion. 4. These results are not consistent with a diminution of TGF function after volume expansion, rather with an enhancement. The latter is best accounted for by vasodilation of preglomerular resistance vessels on volume expansion, a result predicted by calculations from a model based on the serial arrangement of preglomerular and TGF-controlled vascular resistance elements and the established pharmacological actions of ANP. PMID- 2970539 TI - Maximum velocity of shortening related to myosin isoform composition in frog skeletal muscle fibres. AB - 1. The velocity of unloaded shortening (V0), the myofibrillar ATPase activity and the immunoreactivity to two monoclonal antibodies (A1 and A2) that were raised against the myosin heavy chains were studied in single fibres of the anterior tibialis muscle of Rana temporaria. V0 was recorded for the fibre as a whole using the slack-test method. Myofibrillar ATPase activity was determined by means of a quantitative histochemical technique. 2. A highly significant, direct relationship was found to exist between V0 and the myofibrillar ATPase activity recorded in the same single fibres. Both V0 and the myofibrillar ATPase activity changed in proportion to the cross-sectional area of the fibres. 3. Muscle fibres that had first been characterized with respect to V0 and myofibrillar ATPase activity were exposed to monoclonal antibodies A1 and A2. Thin fibres, having relatively low V0 and low myofibrillar ATPase activity, reacted preferentially with A1. Thick fibres, on the other hand, exhibiting relatively high V0 and high myofibrillar ATPase activity, were preferentially stained by A2. A third category of fibres reacted with both A1 and A2. The results support the view that the variability in shortening velocity and myofibrillar ATPase activity that exists among twitch fibres in frog skeletal muscle is based on differences in myosin heavy-chain composition. 4. Attempts were made to elucidate further the previous observation (Edman, Reggiani & te Kronnie, 1985) that the velocity of unloaded shortening (V0) differs along the length of individual muscle fibres. To this end discrete segments (0.5-0.7 mm in length) of intact fibres were delineated by opaque markers of hair that were placed on the fibre surface. The change in length between two adjacent markers (one segment) was recorded photo-electrically while the fibre was released to shorten against a very small load between 2.2 and 2.0 micron sarcomere lengths. In the majority of fibres (eight out of eleven preparations), V0 and myofibrillar ATPase activity exhibited similar patterns of variation along the fibre. Pooled data from thirty-three segments of twelve fibres showed a positive correlation between V0 and myofibrillar ATPase activity (P less than 0.05). 5. The possibility was explored that the myosin isoform composition might vary along the length of an individual muscle fibre. For this purpose bundles of fibres were cross-sectioned at 0.5-1 mm intervals along their entire length and the reactivity to monoclonal antibody A2 was tested at each location.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970540 TI - Effect of peripherally administered atriopeptin III on water intake in rats. AB - 1. Extracellular fluid deficits of 33% were produced in male Long-Evans rats by peritoneal dialysis. The conscious, unrestrained animals were then infused I.V. for 30 min with atriopeptin III at doses of 0.01, 0.1, 0.5 and 1.0 micrograms/min. At 5 min into the infusion, the rats were offered water and subsequent intakes were monitored. Since atrial natriuretic peptide (ANP) causes hypotension, one group of control animals was given an injection of diazoxide sufficient to match this fall in blood pressure. 2. A similar group of rats was prepared for measurement of plasma ANP achieved by infusion. 3. Relative to the saline-infused controls, atriopeptin III did not reduce water intake. Indeed, intake was increased at the highest dose of 1.0 micrograms/min. 4. Relative to the diazoxide controls, water intake was influenced by atriopeptin III in a dose dependent manner, the greatest attenuation being observed at infusions of 0.1 microgram/min. 5. Infusion of atriopeptin III at 0.1 microgram/min caused plasma ANP levels to rise from 252 +/- 21 to 532 +/- 136 pg/ml (n = 9, P less than 0.05) at 15 min. The lowest dose (0.01 microgram/min) caused no detectable increase in plasma levels. 6. It is concluded that, in groups of hypovolaemic rats matched for blood pressure, atriopeptin III caused a dose-related reduction in water intake. PMID- 2970541 TI - Difficulties in laser Doppler measurement of skin blood flow under applied external pressure. AB - During the course of a study of skin blood flow under applied external pressure, it became apparent that decreasing blood flow by loading the skin surface reveals problems that are fundamental to the method of laser Doppler flowmetry. These problems have to do with the fact that the laser Doppler is extremely sensitive to red cell motion of any kind, whether associated with the ordered red cell motion of blood flow or the random red cell motion associated with changes in temperature or vessel occlusion. This effect becomes increasingly important whenever blood flow is compromised (a situation of considerable clinical significance), since the random portion of the signal then becomes significant in comparison with the diminished blood flow. Experiments have been conducted in living animals and with stationary drops of blood which clearly show the importance of these effects with regard to the interpretation of laser Doppler signals. Significant laser Doppler flow signals were repeatedly observed after manipulations which could reasonably be expected to reduce blood flow to zero. PMID- 2970542 TI - Speech perception by congenitally deaf subjects using an electrocutaneous vocoder. AB - Two congenitally profoundly deaf adults were trained to perceive words through the Tacticon 1600 electrocutaneous vocoder, an artificial hearing prosthesis. The subjects learned to identify 50 words during 47 hours (Subject One) and 41 hours (Subject Two) of training, with a 41.6 percent rate of success across all sessions. Both subjects showed consistent error patterns during the training phase. Analysis of these error patterns suggested that they were employing word identification strategies based on some general aspects of tactual patterns. Specific characteristics of the tactual patterns that they appeared to be using included: syllable number, tactual locus of word ending, direction of pattern movement, and position of bursts (/t/, /k/, /d/, for example). Following training, the subjects were tested for their abilities to integrate tactual and aided-auditory cues in word identification. Three conditions of aided-audition alone (A), tactual vocoder alone (TV), and aided-audition with tactual vocoder (TV + A) were used. The stimulus-word list for this phase consisted of the 50 words acquired in tactual vocoder training, and 50 "tactually-new" words, i.e., words that had not been presented to them in tactual vocoder training sessions. They correctly identified 93 percent (Subject One) and 56 percent (Subject Two) more trials in the TV + A condition than in the A condition. Tactually-new vocabulary was correctly identified 78 percent (Subject One) and 50 percent (Subject Two) more often when sensory modalities were combined, than when only aided-audition was used. Subjects identified tactually-new vocabulary better than chance in the TV condition. PMID- 2970543 TI - Spatial, tactile presentation of voice fundamental frequency as a supplement to lipreading: results of extended training with a single subject. AB - An adult with a severe, postlingually-acquired, sensorineural, hearing loss was given 2 hours a week of training in the perception of connected discourse by lipreading, supplemented by voice fundamental frequency (Fo), encoded as locus of vibratory stimulation of the forearm. Three, 3-week blocks of training in the supplemented condition were interspersed with 1-week periods of training by lipreading alone. Training was conducted via a computer-controlled, interactive video system, using a semi-automated connected discourse tracking procedure. Performance was measured as the percentage of words correctly recognized on the first presentation of new sentence material. Scores by lipreading alone averaged approximately 65 percent and remained essentially constant over the 13 weeks of training. Scores under the supplemented condition rose from 65 percent at the beginning of the study to 85 percent at the end. The final supplemented score represented roughly a 50 percent reduction of error rate, when compared with lipreading alone. Performance with the tactile supplement was not as good as with auditorily presented Fo, but was better than has previously been reported in the literature. These data provide evidence to support the notion that subjects can learn to integrate novel tactile codes with the visual stimulus during the lipreading of connected speech. PMID- 2970544 TI - Importance of variables associated with practitioners' estimates of pit and fissure sealant use. AB - Practitioner acceptance of pit and fissure sealants has been slow. To assess the factors that relate to the dentist's decision to use or not to use sealants, a comprehensive survey of 3,504 general dentists and 591 pediatric dentists was undertaken. Response rate to the mailed questionnaire was 37.6 percent (n = 1,193) for general dentists and 61.2 percent (n = 329) for pediatric dentists. Items were grouped into ten scales believed to portray major variables associated with level of sealant use. Level of use, measured as an estimated percent of child patients receiving sealants, served as the dependent variable. A stepwise multiple regression was used to study the relationship between the scales and estimated level of use. Overall, the preventive orientation, opinion, and patient influence scales contributed the most to the explanation of the estimated level of use. These scales, for both general dentists and pediatric dentists, had the strongest simple correlations with sealant use and they also were the three scales that showed up most consistently as the prime scales in the multiple regression analyses. These results suggested that efforts aimed at encouraging a preventive philosophy and improving patient and practitioner opinion and knowledge about sealants should be undertaken. PMID- 2970545 TI - Barrier techniques and hepatitis B vaccine policies in dental components of local health agencies. AB - National health organizations have issued guidelines for barrier control techniques against infectious diseases and for inoculation against hepatitis B. This study surveys the policies of local health agencies regarding the safeguards used in protecting their employees. Local health agencies (N = 201) operating independently of state health departments were mailed questionnaires seeking information regarding glove, mask, and eyewear policies, as well as hepatitis B vaccine policies for new and current employees. Of the 156 programs returning the survey, 127 (76.6%) reported having a clinical component. Many agencies did not require infection control policies to be implemented for every patient. Agencies that had dental directors who have held their positions for fewer than four years were more likely to have requirements for gloves and masks than agencies run by directors with longer tenure. Those agencies that provided dental treatment to inmates at correctional institutions were also more likely to require infection control policies. The data indicate policy inconsistencies among local dental health programs nationwide. Of considerable concern is the fact that infection control was not identified as a priority for the near future. Many local health agencies would benefit from further education in infection control. PMID- 2970546 TI - Time of ovulation in goats (Capra hircus) induced to superovulate with PMSG. AB - The timing of ovulation in feral goats treated with 1200 i.u. PMSG +/- 50 micrograms GnRH was studied by repeated laparoscopy. Experiment 1 established that superovulation began as early as 30 h after withdrawal of progestagen impregnated sponges and was not completed at 54 h if goats received PMSG alone. GnRH synchronized ovulation, leading to 91% of ovulations appearing between 36 and 48 h after sponges were withdrawn. Experiment 2 established that superovulation continued until up to 77 h in goats treated only with PMSG. The stress of repeated laparoscopy appeared to delay or abolish ovulation in some females. The mean (+/- s.e.) ovulation rate was greater in goats treated with GnRH (12.7 +/- 1.3) than in those that received PMSG only (9.7 +/- 1.1; P less than 0.05). Out of 47 of the females in Exp. 1, 43 had one or more corpora lutea at laparoscopy 24 h after withdrawal of progestagen. These early corpora lutea were associated with an increased concentration of plasma progesterone during the periovulatory period. Experiment 3 provided evidence that these corpora lutea arose before the withdrawal of progestagen-impregnated sponges. PMID- 2970547 TI - Facial reconstruction in Down's syndrome. PMID- 2970548 TI - Etodolac, a novel antiinflammatory agent. The syntheses and biological evaluation of its metabolites. AB - The syntheses of five metabolites of the antiinflammatory drug etodolac (1,8 diethyl-1,3,4,9-tetrahydropyrano-[3,4-b]indole-1-acetic acid) are described, viz. 6-hydroxyetodolac, N-methyletodolac, 4-ureidoetodolac, 8-(1'-hydroxy)etodolac, and 4-oxoetodolac. These syntheses were used to confirm the identities of the metabolites. The metabolites themselves, as well as the previously reported metabolite 7-hydroxyetodolac, were tested in a rat adjuvant edema model and in vitro for their capacity to block prostaglandin production in chondrocyte cells. All either were inactive or possessed only marginal activity. The isolation of N methyletodolac and 4-oxoetodolac from human and rat urine, respectively, is also described. PMID- 2970549 TI - 3-[1-(2-Benzoxazolyl)hydrazino]propanenitrile derivatives: inhibitors of immune complex induced inflammation. AB - 3-[1-(2-Benzoxazolyl)hydrazino]propanenitrile derivatives were evaluated in the dermal and pleural reverse passive Arthus reactions in the rat. In the pleural test these compounds were effective in reducing exudate volume and accumulation of white blood cells. This pattern of activity was similar to that of hydrocortisone and different from that of indomethacin. The structural requirements for inhibiting the Arthus reactions were studied by systematic chemical modification of 1. These structure-activity relationship studies revealed that nitrogen 1' of the hydrazino group is essential for activity and must be electron rich, whereas chemical modifications of other sites of 1 had only a modest effect on activity. PMID- 2970550 TI - Synthesis and evaluation of multisubstrate inhibitors of an oncogene-encoded tyrosine-specific protein kinase. 1. AB - The synthesis and testing of potential multisubstrate inhibitors of tyrosine specific protein kinases are described. One of the substrates, ATP, was mimicked by the known kinase inhibitor 5'-[4-(fluorosulfonyl)benzoyl]adenosine, which was covalently linked via the sulfonyl moiety to tyrosine mimics. The resulting multisubstrate inhibitors were tested for their ability to inhibit the transfer of phosphate from ATP to a protein acceptor by p60v-abl, the tyrosine kinase encoded by the transforming gene (v-abl) of the Abelson murine leukemia virus (A MuLV). Although the series of inhibitors displayed moderately potent activity (IC50 values as low as 19 microM), the absence of large effects produced by modification of the tyrosine mimic suggests that they do not behave as multisubstrate inhibitors but bind primarily through the adenosine moiety common to all the inhibitors. This interpretation is strengthened by the finding that the inhibitors lack specificity, inhibiting a serine kinase at comparable concentrations. PMID- 2970551 TI - Synthesis and in vitro activity of various derivatives of a novel thromboxane receptor antagonist, (+/-)-(5Z)-7-[3-endo-[(phenylsulfonyl)amino]bicyclo[2.2.1] hept-2-exo-yl]heptenoic acid. AB - Several sulfonyl derivatives (13a-t) of (+/-)-(5Z)-7-(3-endo aminobicyclo[2.2.1]hept-2-exo-yl)heptenoic acid (VI) were synthesized via its methyl ester 10. Sulfonylation of 10 with 11a-t followed by saponification yielded 13a-t. Inhibitory concentrations (IC50) of the corresponding sodium salts 14a-t for platelet aggregation were measured with rat washed platelets (WP) and rabbit platelet-rich plasma (PRP). IC50 values of some derivatives for contraction of the rat aorta were also measured. The IC50 values for rat WP increased from 2.9 to 26 nM in the order of 14a, 14c, 14d, and 14b for derivatives with an arylsulfonyl residue, depending on the number of of intervening methylene groups. Methyl derivative 14e exhibited a higher IC50 value than n-hexyl derivative 14f. Substitution with a p-methyl, p-fluoro-, or p-chloro group in 14a retained or slightly reduced its IC50 value, while a p-n-pentyl or p oxycarbonyl group augmented it significantly. The representative 14a suppressed (15S)-15-hydroxy-11,9-(epoxymethano)prosta-5(Z),13(E)-dienoic acid (U-46619) induced aggregation of human WP with an IC50 value of 7.7 nM, which corresponds well to the IC50 value of 3 nM obtained for each displacement by 14a of [3H]-U 46619 or (5Z,15 xi)-9 alpha, 11 alpha-(dimethylmethano)-15-hydroxy-16-(3 [125I]iodo- 4-hydroxyphenyl)-17,18,19,20-tetranor-13-aza-11a-carbathrombo-5-en oic acid [( 125I]-PTA-OH) bound to human WP. Synthesis of thromboxane A2 (TxA2) in human WP stimulated by thrombin was not inhibited by 14a at a concentration up to 10 microM. From these observations, the corresponding acid 13a (S-145) was concluded to be a potent TxA2 receptor antagonist. PMID- 2970552 TI - Characterization of a doubly mutant derivative of the lambda PRM promoter. Effects of mutations on activation of PRM. AB - The mutation, prmE37, located at -14 in the PRM promoter of bacteriophage lambda, reduces PRM function dramatically both in vitro and in vivo. In a search for second-site revertants of prmE37, we isolated a double mutant that exhibits a partially restored Prm+ phenotype. The second-site mutation (at -31) is identical to the mutation prmup-1. The activity of the doubly mutant (pseudo-revertant) promoter, prmE37prmup-1, was investigated in vivo using a PRM-lacZ fusion phage and found to be intermediate between that of prmE37 and wild-type PRM. However, the relative strength of the prmE37prmup-1 promoter was greater than expected following superinfection of a lambda lysogen. Since nalidixic acid was found to preferentially inhibit transcription from the doubly mutant promoter under these conditions, we suggest that DNA supercoiling favors activation of this promoter by repressor. In runoff transcription assays in the absence of repressor, the activity of wild-type PRM and the doubly mutant promoter were the same. However, while addition of repressor significantly stimulated wild-type PRM, it had little or no effect on the activity of the doubly mutant promoter. Values of KB, the equilibrium constant for formation of closed complexes, and kf, the rate constant for isomerization of closed to open complexes, were determined in abortive initiation assays, and the product of kfKB was used as a measure of promoter strength. The results of these assays are in agreement with those obtained in runoff transcription assays. In the absence of repressor, values of kfKB for the doubly mutant promoter and wild-type PRM are the same; however, tau obs, the time required for open complex formation, is significantly greater for the double mutant than for wild-type PRM at all RNA polymerase concentrations used for the abortive initiation analysis. In the presence of repressor, the doubly mutant promoter is stronger than the prmE37 promoter, but much weaker than wild-type PRM. This is due to the fact that kf for the doubly mutant promoter is increased 2.5-fold by repressor, but KB is reduced to the same extent. These two effects counteract each other, so that repressor has no net effect on the strength of the prmE37prmup-1 promoter in vitro. In contrast, repressor increases kf for wild type PRM eightfold and increases overall promoter strength (KBkf) nearly fivefold. In the presence of repressor, the effects of the two mutations, prmE37 and prmup-1, on kf are independent. This observation is discussed in relation to revised models for open complex formation. PMID- 2970554 TI - Phase II study of intravenous menogaril in patients with advanced breast cancer. AB - Menogaril was administered to 40 patients with advanced breast cancer who had not received anthracycline drugs previously. The drug was given iv as a 2-hour infusion, repeated every 4 weeks, at doses of 200 mg/m2 and 160 mg/m2 in good risk and poor-risk patients. The overall response rate was 22% in patients with no prior chemotherapy and 10% in patients previously exposed to chemotherapy. Leukopenia was generally moderate and predictable. Phlebitis and erythema along the vein injected occurred in 34% and 17% of the cases, respectively. Menogaril is an active drug used in the treatment of patients with advanced breast cancer who have not had prior systemic therapy. PMID- 2970553 TI - Crystals of an ATPase fragment of bovine clathrin uncoating ATPase. AB - A 44,000 Mr amino-terminal, clathrin-independent ATPase fragment of the bovine clathrin uncoating ATPase has been crystallized in a form suitable for X-ray diffraction studies. The crystals are orthorhombic, space group P2(1)2(1)2(1), a = 145.3 A, b = 65.0 A, c = 46.9 A, with one protein molecule per asymmetric unit (1 A = 0.1 nm). PMID- 2970555 TI - Randomized trial comparing aminoglutethimide with high-dose medroxyprogesterone acetate in therapy for advanced breast carcinoma. AB - A total of 218 postmenopausal patients were entered in a prospective randomized trial comparing aminoglutethimide (AG) and high-dose medroxyprogesterone acetate (MPA) as second-line hormonal therapy for advanced breast carcinoma. All responses were assessed by the criteria of the International Union Against Cancer. The response rates were 27% (29 of 106 patients) for AG and 31% (35 of 112) for MPA, but if stabilization of previously progressive disease is included, then the overall response rates were 51% (54 of 106) and 54% (61 of 112) for patients receiving AG or MPA, respectively. There was no difference in response to the two drugs at any site of disease, and the durations of response and survival were identical for the two drugs. The time to response was significantly shorter for patients treated with MPA (median, 8.7 wk) than for those treated with AG (median, 15.3 wk) (chi 2 = 9.96, 1 df, P = .0016). The percentage of patients experiencing toxic effects was equivalent in both arms, although the patterns and time courses of these effects were different. PMID- 2970556 TI - The neurohumoral response to burn injury in patients resuscitated with hypertonic saline. AB - Fourteen adult patients (mean age, 35 yrs) with 20-60% total body surface area (TBSA) burns (mean, 35%) were resuscitated using hypertonic sodium lactate (HSL: sodium = 250 mEq/L). Plasma concentrations of atrial natriuretic peptide (ANP), arginine vasopressin (AVP), angiotensin II (A-II), epinephrine (E) and norepinephrine (NE) were measured on admission and for 7 days following burn injury. Serum sodium concentrations and osmolalities were lowest on admission, and were persistently elevated following HSL resuscitation. Plasma AVP levels were highest on admission and correlated with the size of the burn injury. Between days 4 and 5 plasma ANP levels rose while plasma AVP levels returned to normal. Plasma concentrations of AVP and ANP did not correlate with serum osmolality or serum sodium concentrations on admission or after HSL resuscitation. Plasma levels of A-II, NE and E were elevated throughout the 7-day period and were unrelated to the size of the burn. PMID- 2970558 TI - Percutaneous transluminal coronary angioplasty for elderly patients. AB - To examine the clinical efficacy of percutaneous transluminal coronary angioplasty in elderly patients, 350 consecutive procedures of coronary angioplasty were reviewed by dividing the patients into two groups: 142 cases of elderly patients whose ages were 65 years or older and 208 cases of younger patients. The primary success rate was satisfactory in both groups (86.6% in the elderly patients and 88.5% in the younger patients) and frequency of complication was acceptable (3.1% vs 2.9%) in both groups. Although restenosis tended to occur more frequently in the elderly patients (36.8% vs 27.1%), most of the lesions were re-dilated. These indicate that coronary angioplasty can be extended to elderly patients. The primary success rate started to decrease with circumflex arteries at age 70 years, and a similar trend was seen with right coronary arteries at age 65 years. Careful selection of the patients for coronary angioplasty as well as detailed analysis of the coronary anatomy and aorta are required to obtain clinical success and to prevent complications. PMID- 2970557 TI - Lack of response to recombinant hepatitis B vaccine in nonresponders to the plasma vaccine. AB - Yeast recombinant hepatitis B vaccine was administered to 25 nonresponders to the plasma-derived hepatitis B vaccine. After three 10-micrograms doses, nine subjects (36%) produced levels of antibodies to hepatitis B surface antigen (anti HBs) of less than 2.1 sample ratio units (SRU) (nonresponders), and five (20%) developed anti-HBs of 2.1 to 9.9 SRU (hyporesponders); anti-HBs levels of 10 SRU or greater were detected at least once in 11 vaccinees (44%), but by the sixth and 12th months after the last vaccination, only three and one of these "responders," respectively, still maintained anti-HBs values of 10 SRU or greater. In these 25 subjects HLA subtyping showed a high prevalence of DR7, B8, and the combinations of DR3 and DR7 and DR4 and DR7. Our findings indicate that the yeast recombinant hepatitis B vaccine was not effective in eliciting a sustained anti-HBs response in nonresponders to the plasma hepatitis B vaccine. PMID- 2970559 TI - [Cytokines and receptors--their functions, structure and cloning of code genes. Interleukin-1]. PMID- 2970560 TI - [Percutaneous angioplasty in aortic coarctation]. PMID- 2970561 TI - [Thrombocyte aggregation, levels of thrombocyte factor 4 and beta-thromboglobulin in the blood plasma of patients with unstable stenocardia and acute myocardial infarct]. PMID- 2970562 TI - Marked acceleration of the autoimmune disease in MRL-lpr/lpr mice by the influence of the Yaa gene from BXSB mice. AB - MRL-lpr, Yaa males were developed by the transfer of the Yaa gene from BXSB males to the MRL-lpr strain. The early-onset autoimmune disease of MRL-lpr males was accelerated further by the action of Yaa. All the serological parameters of autoimmune disease examined, i.e., anti-DNA antibodies, rheumatoid factors and circulating immune complexes were elevated earlier in MRL-lpr, Yaa males than in MRL-lpr males. Consequently, the MRL-lpr, Yaa males showed the clinical signs of an earlier and more rapidly progressive glomerulonephritis as compared with MRL lpr males. The lifespan of MRL-lpr, Yaa males was shortened by about 50% in comparison to MRL-lpr males, with 90% of MRL-lpr, Yaa males dead by about 4 months of age. The clinical features and overall intensity of the autoimmune responses in the terminal stages appeared to be similar in both MRL-lpr, Yaa and MRL-lpr males. Thus, Yaa appeared merely to accelerate the disease course of MRL lpr males, without altering the essential nature of the disease. PMID- 2970563 TI - Cardiac architectural changes with hypertrophy induced by excess growth hormone in rats. AB - We have shown that excess growth hormone (GH) causes left ventricular hypertrophy in rats. To further characterize cell growth of this model, combined light and electron microscopic morphometric studies were done on 5- and 1-micron sections of the perfusion fixed left ventricle (LV) in 6 control (C) and 5 GH-stimulated rats. Total ventricular weight, LV weight and right ventricle weight were increased 45, 46, and 43%, respectively, in the rats after a few weeks of GH stimulation. However, neither the ratio of LV to body weight, nor right ventricle to body weight were significantly different between the two groups. Nuclei (n)/volume decreased by 25% in GH due to a decreased number of n/area (819 +/- 80 versus 718 +/- 52/mm2, p less than 0.05) and an increased n length (16.9 +/- 1.7 versus 19.7 +/- 0.5 micron, p less than 0.01). As the volume fraction of myocytes was the same in GH and C, the volume of myocytes/n increased (16.5 +/- 1.9 in C to 20.8 +/- 1.1 x 10(3) micron3 in GH, p less than 0.01). Since no change was found in cross-sectional area of myocytes (242 +/- 26 in C versus 244 +/- 23 micron2 in GH, p greater than 0.50) cell length/nucleus increased by 26% (68.5 +/ 8.6 in C versus 86.0 +/- 7.8 micron in GH, p less than 0.01). The average area of mid-LV wall increased by 35% in GH compared with C (p less than 0.01), but wall thickness was unchanged (2.1 +/- 0.4 in C versus 2.2 +/- 0.2 mm in GH, p greater than 0.20) as LV cavity radius increased (1.85 +/- 0.5 in C to 2.50 +/- 0.6 mm in GH, p = 0.06). With decreased developed pressure of LV in GH, systolic circumferential wall stress was unchanged (55.3 +/- 21.1 in C versus 57.2 +/- 22.5 x 10(3) dyne/cm2 in GH, p greater than 0.50). We conclude that left ventricular hypertrophy of this model is associated with cavity dilation and increased myocyte cell length; this adaptive response tends to minimize the effects of increased volume load on wall stress. PMID- 2970564 TI - Mitogenic activity of high molecular polysaccharide fractions isolated from the Cupressaceae Thuja occidentale L. I. Macrophage-dependent induction of CD-4 positive T-helper (Th+) lymphocytes. AB - The arborvitae or Thuja occidentale L., one of the Cupressaceae, has rarely been investigated until now. Several authors have demonstrated that allopathic extracts of this plant could be used as strong antiviral agents directed against plant and animal viruses. Polysaccharide fractions with molecular weights ranging between 20,000 and 1,000,000 and higher have been isolated from the aqueous alkaline extract of the herbal parts of T. occidentale L. by ethanol precipitation and fractionation by ultrafiltration. High molecular subfractions of Thujapolysaccharides (TPS) proved to be highly mitogenic on peripheral blood leukocytes. It was demonstrated by the alcalic immune phosphatase-antiphosphatase and Pappenheim staining methods that the mitogenic and cluster-forming activity of TPS cause T cell induction, in particular, of CD 4-positive T-helper/inducer cells as opposed to B cells. The CD-4+ T-helper/inducer cell induction is connected to an increased production of IL-2. The cluster-forming ability and mitogenity of TPS correlates well with the 3H-thymidine uptake and seems to be IL 1 and IFN-gamma dependent as could be shown by blocking the mitogenic effect using anti-IL-1- and anti-IFN antibodies. Whether it is possible to use these polysaccharide fractions as an adjuvant in the therapy of immune deficiency syndromes and cancer must now be further investigated. PMID- 2970565 TI - Left ventricular morphological changes in primary hypertension: echocardiographic evidence for two patterns of hypertrophy. PMID- 2970566 TI - Transformation of the [3H]aldosterone-type I receptor complex to a DNA-binding species. AB - For most steroid receptor complexes, the transformation to a DNA-binding species can be achieved readily in vitro by incubation at elevated temperatures and/or salt concentrations. Although the aldosterone-Type I receptor complex forms a clear exception to this generalization, a marked increase in its transformation can be achieved by incubation with the chaotropic anion, thiocyanate. Time and concentration-response analyses with brain cytosol revealed that over 40% of the complexes were retained in DNA-cellulose assays after a 15 min pre-incubation at 0 degree C with 100 mM thiocyanate. As expected, molybdate prevented this transformation; however, in contrast to results with heat- and/or salt-induced transformation of other steroid receptors, the molybdate effect was only partially removed by gel filtering the cytosol prior to thiocyanate addition. Thiocyanate-induced transformation should prove useful in the biochemical characterization and purification of non-transformed and transformed aldosterone Type I receptor complexes. PMID- 2970568 TI - Renal responses to atrial natriuretic factor in hydrated and hydropenic dogs. AB - Atrial natriuretic factor (ANF 101-126) was compared to the standard diuretics, furosemide and hydrochlorothiazide, and to the vasodilator, acetylcholine in hydrated and dehydrated anesthetized dogs. ANF 101-126 (20 pmole/kg/min, ira) modestly reduced solute-free water clearance in water-loaded dogs and slightly lowered free water reabsorption in dehydrated animals. This pattern of responses most closely resembled those produce by 10 mg/kg, ira of the distally-acting diuretic, hydrochlorothiazide and a natriuretic dose of acetylcholine (2.5 micrograms/kg/min, ira). In contrast, the loop diuretic, furosemide (1 mg/kg, ira) drastically suppressed both free water clearance and reabsorption. ANF 101 126 produced changes in free water handling which were not readily distinguishable from those induced by either hydrochlorothiazide, a distally acting diuretic, or acetylcholine, a vasodilator. PMID- 2970567 TI - Changes in lymphocyte subsets and macrophage functions from high, short-term dietary ethanol in C57/BL6 mice. AB - Chronic administration of a diet containing 7% ethanol (36% of total calories) for 8 days to male C57/BL6 mice resulted in significant changes in functioning of macrophages. Peritoneal exudate macrophages from the ethanol-fed mice released more tumor cell cytotoxic materials upon culturing in vitro than cells from controls. However, peritoneal exudate cells continued to respond to exogenous beta carotene in vitro to produce additional cytotoxic materials. Phagocytosis of sheep red blood cells in vitro was suppressed in cells from ethanol treated mice. The number of splenic lymphocytes of various subsets was significantly changed by the ethanol exposure. Total T cells and T suppressor cells were lower, with a significant decrease in B cells containing IgM on their surface. The percentage of spleen cells showing markers for macrophage functions and their activation were significantly reduced. It is concluded that short-term chronic consumption of dietary ethanol, which was sufficient to produce physical dependence, results in significant alterations in lymphocyte subtypes and suppression of some macrophage functions. PMID- 2970569 TI - Studies on the effect of 1-deoxynojirimycin on the release of albumin, sialyltransferase and alpha 1 acid glycoprotein from liver slices from normal and inflamed rats. AB - Liver slices from control and 24hr inflamed rats were incubated for up to 20hr with 5mM 1-deoxynojirimycin (DN), an inhibitor of the processing glucosidases. The amounts of albumin and alpha 1-acid glycoprotein (AGP) and the activities of sialyltransferase were determined in liver and medium. The presence of DN significantly inhibited the release of AGP and sialyltransferase. The inhibitory effect of DN was most pronounced with slices from inflamed rats. Secretion of albumin was not inhibited. Incorporation studies with labelled leucine and mannose showed that the inhibitor did not significantly affect protein synthesis, but it did inhibit mannose incorporation into AGP and sialyltransferase. The results show that DN inhibits the secretion of acute phase AGP and sialyltransferase in liver slices and further suggests that sialyltransferase is a glycoprotein. PMID- 2970570 TI - A sensitive radioimmunoassay of alpha human atrial natriuretic polypeptide using monoclonal antibody recognizing human form ring structure. AB - A monoclonal antibody (C351) against alpha human atrial natriuretic polypeptide (alpha hANP) recognizing human form ring structure was established and applied to a radioimmunoassay of plasma alpha hANP. The minimum detectable amount in terms of 10% radioligand displacement relative to zero dose were 0.28 fmol/tube, corresponding to 0.7 fmol/ml in plasma after extraction using Sep-Pak C18 cartridges. When the mean plasma levels at recumbent position in fasted morning were compared in 10 young (less than 30 years) and 10 elderly (greater than or equal to 50 years) healthy subjects taking normal sodium diet, it was slightly higher in the latter (3.2 +/- 0.4 vs 4.7 +/- 0.5 fmol/ml, mean +/- SE, p less than 0.05). After i.v. infusion of hypertonic saline (2.5% NaCl) at a rate of 0.24 ml/kg/min for 20 min in 6 normal subjects (26 to 35 years), it was increased from 4.1 +/- 0.4 to 5.9 +/- 0.7 fmol/ml (p less than 0.01). In 6 patients with essential hypertension (34 to 57 years), it was elevated with high salt intake, i.e. 3.3 +/- 0.3, 3.9 +/- 1.03 and 7.6 +/- 1.5 fmol/ml under 34, 170 and 340 mEq NaCl/day for 7 days, respectively. From these results, the radioimmunoassay of plasma IR-alpha hANP using MAb C351 seems to be quite suitable to detect rather small changes at low plasma concentrations and to investigate a physiological importance of alpha hANP in man. PMID- 2970571 TI - Effect of kynurenine loading on quinolinic acid production in the rat: studies in vitro and in vivo. AB - In vitro and in vivo techniques were used to examine the production and subsequent fate of the endogenous excitotoxin quinolinic acid (QUIN) following administration of its bioprecursor L-kynurenine (KYN). Incubation of liver slices in the presence of 10-1000 microM KYN resulted in a dose- and time-dependent release of QUIN into the incubation medium. Less than 15% of total QUIN produced was recovered from the tissue. In vivo experiments, performed with a microdialysis probe inserted in the jugular vein of anesthetized rats, showed that injection of KYN (20-600 mg/kg, i.v.) causes rapid and dose-dependent increases in the serum level of QUIN. Peak QUIN concentrations in serum dialysates were reached 75 minutes following KYN administration. Longer lasting increases were detected following the administration of pyrazinamide (20 mg/kg, i.p.), an indirectly acting stimulator of QUIN biosynthesis in the periphery. The data demonstrate the feasibility of assessing the mechanisms of QUIN production and disposition in experimental paradigms which can be expected to allow insights into the function and possible dysfunction of QUIN in the brain. PMID- 2970572 TI - The reproducibility of resting and post exercise plasma beta-endorphins. AB - This investigation examined the reproducibility of resting and post exercise plasma beta-endorphin levels. Twenty subjects (10 men and 10 women) had their resting endorphin levels measured under controlled conditions on four separate occasions. Concomitantly, the endorphin response of eight trained runners completing three similar ten mile runs was also determined. For the resting data, there was no significant overall variation among trials, but the intra-subject variability was substantial; the within subject variance was 6.16, and it corresponded to an intra-class reliability coefficient of r = 0.239. No gender effect was noted for the average beta-endorphin values for the four occasions (men = 4.6 +/- 1.7; women = 4.4 +/- 2.1 pM/l); however, the males' within-subject variance of 8.548 (r = 0.080) was significantly larger than that of 3.719 (r = 0.485) for females. Of the runners, one outlier subject had a uniquely high average beta endorphin level of 85.67. Analysis including and excluding the outlier subject yielded within-subject variances of 29.61 (r = 0.960) and 34.47 (r = 0.176), respectively; variances for differences in confidence limits for random variation, they must exceed 7 pM/l at rest, 17 pM/l post exercise, and 20 pM/L difference from rest to post exercise. PMID- 2970573 TI - [The role of the nurse and midwife in the comprehensive prevention of health disorders in mother and child]. PMID- 2970574 TI - [Characteristics of the work of a nurses' council in a district hospital]. PMID- 2970575 TI - [Work experience of the nurses' council of the N. A. Semashko Gorki Province Clinical Hospital]. PMID- 2970576 TI - [Ways to rationalize the work of nurses in a central district hospital]. PMID- 2970577 TI - cGMP-dependent protein kinase activation in intact tissues. PMID- 2970578 TI - Children who look different. PMID- 2970579 TI - T4 cell receptor distortion in acquired immune deficiency syndrome. AB - Among the possible affects of the HIV virus on T-cells in Acquired Immune Deficiency Syndrome is the cell membrane distortion of the T4 cell protein receptor for Genetically Related Macrophage Factor (GRF). The integrity of this specific T-cell receptor takes on profound significance due to its relationship to soluble antigen. Alpha-tocopheral has been shown to assist in the maintenance of cell membrane structure and may have application in a strategy of membrane restoration. Adoptive Transfer, a method of immune cell alteration applicable in anti-tumor therapy, may play a role in the mechanics of such a strategy. PMID- 2970580 TI - Night terrors and sudden unexplained nocturnal death. AB - A high incidence of sudden unexplained nocturnal deaths has been reported among young Asian males. These deaths are known as Pokkuri in Japan, Bangungut in the Philippines and Sudden Unexplained Nocturnal Death in the United States. Post mortem analysis has demonstrated cardiac conduction defects in many of the victims. Careful review of the terminal events surrounding these deaths suggests that the victims suffered from night terrors. Night terrors are a sleep disorder characterized by vocalization, motor activity, a nonarousable state, and severe autonomic discharge. The proposed recognition of both night terrors and cardiac anomalies in these patients offers a pathophysiologic mechanism for their sudden death. PMID- 2970581 TI - [Dicrocoelium lanceatum infestation in humans (based on autopsy data)]. PMID- 2970583 TI - Of golden eggs. PMID- 2970582 TI - [Significance of a combined pinworm and beef tapeworm infestation in the transmission of oncospheres to cattle]. PMID- 2970584 TI - Economic death--and life. PMID- 2970585 TI - [Carcinoma of Papilla Vater with celiac axis stenosis--application of percutaneous transluminal angioplasty]. AB - In a case of carcinoma of Papilla of Vater with angiographically proved celiac axis stenosis, PTA (Percutaneous transluminal angioplasty) was performed to restore sufficient blood flow, followed by standard pancreaticoduodenectomy. This would be the first case to have PTA applied to the cancer-bearing celiac axis stenosis. The patient was 58 year old female, presented with acute abdominal pain and vomiting. DIC revealing protruding defect in the choledochus and hypotonic duodenography showed elevated tumor at Papilla of Vater. Aortography revealed 75% circumferential stenosis at the celiac artery and superior mesenteric angiography visualized all the branches of the celiac axis via dilated pancreaticoduodenal arcades. PTA with Gruntzig Dilaca (4.3Fr), 5 atm for 60 seconds serially every two min for 4 times was performed with the immediate relief of the stenosis down to 25%. Intraoperatively 70-80ml/sec flow confirmed, pancreaticoduodenectomy was done without sacrificing curability. PTA would be superior strategy to restore sufficient blood flow in cases with GI anastomoses, compared with other bypass or reimplantation surgery. PMID- 2970586 TI - Dopamine D2 receptor binding subunits of Mr congruent to 140,000 and 94,000 in brain: deglycosylation yields a common unit of Mr congruent to 44,000. AB - The ligand-binding subunit of the porcine striatal dopamine D2 receptor was identified by photoaffinity labeling with [125I]N-azidophenethylspiperone ([125I]NAPS). Upon photolysis, [125I]NAPS covalently incorporated into a broad band of apparent Mr congruent 140,000 with an appropriate pharmacological profile for D2 receptors as assessed by autoradiography after sodium dodecyl sulfate polyacrylamide gel electrophoresis. Smaller subunits of apparent Mr congruent 94,000 and 34,000 were specifically labeled by [125I]NAPS with an appropriate D2 receptor profile and were similar to the major ligand-binding subunits of photoaffinity-labeled canine striatal D2 receptors. Photoaffinity labeling in the absence or presence of multiple protease inhibitors did not alter the migration pattern of the Mr congruent to 140,000/94,000 subunits upon denaturing electrophoresis in either the absence or presence of thiol-reducing/alkylating reagents. In order to investigate the possible basis for the existence of these high molecular weight forms of the D2 receptor, we assessed the carbohydrate nature of photolabeled D2 ligand-binding subunits by the use of lectin affinity chromatography and specific exo- and endoglycosidase treatments. Both photoaffinity-labeled D2 receptor proteins from porcine striatum (Mr congruent to 140,000 and 94,000) were glycoproteins as indexed by their absorption and specific elution from wheat germ agglutinin lectin resins. The exoglycosidase neuraminidase altered the electrophoretic mobility of both the Mr congruent to 140,000 and 94,000 labeled subunits to a single band of apparent Mr congruent to 51,000. Prior removal of sialic acid residues did not alter the reversible binding characteristics of [3H]spiperone to D2 receptors. Complete removal of receptor-associated N-linked carbohydrate by the endoglycosidase glycopeptidase F (peptide-N4[N-acetyl-beta-glucosaminyl]asparagine amidase) produced a further increase in the mobility of the Mr congruent to 51,000 subunit to apparent Mr congruent to 44,000. The porcine Mr congruent to 34,000 photolabeled peptide is an N-linked glycoprotein as assessed by lectin affinity chromatography and susceptibility to digestion by glycopeptidase F to a peptide of apparent Mr congruent to 23,000.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2970587 TI - Bepridil and cetiedil reversibly inhibit thyroid hormone stimulation in vitro of human red cell Ca2+-ATPase activity. AB - Thyroid hormone (10(-11) to 10(-10) M) stimulates plasma membrane Ca2+-ATPase activity in vitro in various tissues, including the human red cell (RBC), by a calmodulin-requiring mechanism. Bepridil and cetiedil are Ca2+ antagonists with an intracellular (calmodulin-antagonist) site of action, as well as an effect on the calcium channel in excitable tissues. We have studied the actions of bepridil and cetiedil on Ca2+-ATPase in a channel-free membrane (RBC) to determine effectiveness of these agents as inhibitors of thyroid hormone action on the enzyme. Dose-response studies showed that thyroid hormone stimulation of Ca2+ ATPase activity in vitro was significantly inhibited by as little as 2 x 10(-5) M bepridil and cetiedil. IC50 values of bepridil and cetiedil for thyroid hormone response of the enzyme were 5 x 10(-5) and 2 x 10(-5) M, respectively, whereas IC50s of these agents for enzyme activity in the absence of thyroid hormone were both 10(-4) M. Progressive addition of purified rat testis calmodulin in vitro (10-150 ng calmodulin/mg membrane protein) restored hormone responsiveness in the presence of bepridil and cetiedil. Binding of labeled thyroid hormone by RBC membranes was unaffected by bepridil and cetiedil (up to 2 x 10(-4) M). Thus, bepridil and cetiedil are Ca2+ antagonists that reversibly inhibit thyroid hormone action on human RBC Ca2+-ATPase by a calmodulin-dependent mechanism. Thyroid hormone effect on Ca2+-ATPase is more susceptible to bepridil and cetiedil inhibition than is basal enzyme activity. PMID- 2970588 TI - [Orofacial re-equilibration using a palatal splint in Down's syndrome]. PMID- 2970589 TI - Solubilization and kinetic characterization of mitochondrial adenosine triphosphatase from Leishmania donovani promastigotes. AB - Oligomycin-sensitive particulate ATPase (MB ATPase) from L. donovani promastigotes was solubilized by chloroform treatment. Polyacrylamide gel electrophoresis revealed several protein bands, with the major one possessing ATPase activity. The solubilized enzyme had Mg2+-ATPase and Ca2+-ATPase but no K+ dependent alkaline phosphatase activity. The Mg2+-ATPase activity was stimulated by monovalent cations and was not sensitive to oligomycin. Hence it is referred to as F1 ATPase. It had optimum activity at pH 7.6 and 30 degrees C. The Arrhenius plot for MB ATPase was biphasic with activation energies (Ea) of 16.2 and 3.4 kcal mol-1, while F1 ATPase exhibited a linear plot with Ea = 10.1 kcal mol-1. Lineweaver-Burk plots were biphasic with Km values of 0.17 and 1.25 mM for MB ATPase and 0.18 and 1.33 mM for F1 ATPase. The enzyme could be preserved at 15 degrees C in Tris-SO2-(4)-EDTA-ATP-glycerol (t1/2 = 20 days). PMID- 2970590 TI - Ventricular activation of the atrial natriuretic factor gene in acute myocardial infarction. PMID- 2970591 TI - Adrenal transplants for Huntington's disease? PMID- 2970592 TI - Intrathymic deletion of self-reactive cells prevented by neonatal anti-CD4 antibody treatment. AB - T-cell differentiation in the thymus involves the coordinate expression of genes encoding the alpha and beta chains of the major histocompatibility complex restricted heterodimeric antigen receptor (TCR) complex, as well as other functionally important molecules such as CD4 and CD8. The repertoire of TCR expressed by T cells is generally thought to be influenced by positive and/or negative selection events occurring when TCRs on developing T cells interact with self-antigens and major histocompatibility complex components. Using a model system in which specific antigen-reactive cells can be monitored by virtue of their preferential expression of certain TCR beta-chain variable (V beta) domains, it has been shown that self-reactive T cells are clonally deleted during development. We report here that clonal deletion of V+ beta 6 cells in Mlsa mice can be prevented by in vivo neonatal administration of monoclonal antibodies directed against CD4. Furthermore, as anti-CD4 monoclonal antibody treatment resulted in the reappearance of V+ beta 6 cells in the mature CD8+ T-cell subset, it is likely that clonal deletion acts on the CD4+CD8+ thymocyte subset and that this subset is an intermediate stage in the differentiation pathway of both CD4+ and CD8+ T-cell lineages. PMID- 2970593 TI - Thymic major histocompatibility complex antigens and the alpha beta T-cell receptor determine the CD4/CD8 phenotype of T cells. AB - T-cell receptors and T-cell subsets were analysed in T-cell receptor transgenic mice expressing alpha and beta T-cell receptor genes isolated from a male specific, H-2Db-restricted CD4-8+ T-cell clone. The results indicate that the specific interaction of the T-cell receptor on immature thymocytes with thymic major histocompatibility complex antigens determines the differentiation of CD4+8+ thymocytes into either CD4+8- or CD4-8+ mature T cells. PMID- 2970595 TI - [Relation between low backache and housekeeping habits in women in The Netherlands and Japan]. PMID- 2970594 TI - Transfer of a functional human immune system to mice with severe combined immunodeficiency. AB - The pressing need for a better experimental system for AIDS research has brought into sharp focus the shortcomings of available animal models and the practical and ethical limitations of studies of immune responses and viral pathogenesis in humans. Current studies of the human immune responses are limited to relatively restrictive in vivo experiments and several in vitro systems that, although useful, allow only short-term studies and support responses to a few antigens. Neither model is particularly amenable to studies of the pathogenesis of diseases of the immune system. We report here that injection of human peripheral blood leukocytes (PBL) can result in the stable long-term reconstitution of a functional human immune system in mice with severe combined immunodeficiency (SCID). Human PBL transplanted to SCID mice increase in number and survive for at least six months; reconstituted mice show spontaneous secretion of human immunoglobulin and a specific human antibody response is induced following immunization with tetanus toxoid. All of the major cell populations present in PBL are found in the lymphoid tissue and blood of SCID recipients, although the relative proportions of B cells, T-cell subsets and monocytes/macrophages in long term recipients differ from those found in normal PBL and, in mice transplanted with 50 x 10(6) or more PBL from Epstein-Barr virus (EBV)-seropositive donors, EBV-positive B-cell lymphomas often develop. Our results suggest that xenogeneic transplantation of human lymphoid cells into SCID mice may provide a useful model for the study of normal human immune function, the response of the immune system to pathogenic agents and early events in lymphomagensis. PMID- 2970596 TI - Campaigning for compensation. PMID- 2970597 TI - Who needs nurses? PMID- 2970598 TI - [The locked-in syndrome--experience dimensions and possibilities for an enlarged communication system]. PMID- 2970599 TI - Analysis of left-ventricular changes associated with chronic hemodialysis. A noninvasive follow-up study. AB - To assess the reasons for the frequent cardiovascular complications in patients with end-stage renal disease (ESRD), 61 out of 131 normotensive ESRD patients originally examined (mean ESRD duration: 71 +/- 41 months) were followed over 2.5 years by echo-, electro- and mechanocardiography. Clinical and biochemical parameters were comparable. The prevalence of pericardial effusion (3%), pericardial thickening (14%), aortic valve sclerosis (14%) and mitral valve anulus sclerosis (12%) was unchanged. The interventricular septum diameter (14.3 +/- 3.0 vs. 16.4 +/- 3.4 mm), index of left-ventricular (LV) wall asymmetry (1.25 +/- 0.30 vs. 1.52 +/- 0.36) and left atrial diameter (38.3 +/- 5.4 vs. 42.6 +/- 3 mm) increased (p less than 0.001). The LV end-systolic diameter decreased slightly (35.8 + 6.3 vs. 34.2 +/- 6.4 mm; p less than 0.05), with no significant changes for end-diastolic diameter (50.4 +/- 6.3 vs. 49.3 +/- 6.1 mm), muscle mass index (189 +/- 57 vs. 197 +/- 50 g/m2), stroke volume (86.1 +/- 26.2 vs. 85.7 +/- 26.7 7 ml/m2) and fractional shortening (29.1 +/- 7 vs. 30.8 +/- 8.6%). We conclude that the predominant finding in ESRD is an LV hypertrophy progressing towards an asymmetric septum hypertrophy, while the increase of the primarily enlarged left atrial diameter over 30 months reflects a further deterioration of the diastolic LV dysfunction. PMID- 2970601 TI - [Relation, in Raynaud's phenomenon, of clinical severity and severity as disclosed by 2 methods of non-invasive vascular diagnosis]. PMID- 2970600 TI - The minor symptoms of increased intracranial pressure: 101 patients with benign intracranial hypertension. AB - Of 101 patients with benign intracranial hypertension not related to vasculitis, neck stiffness occurred in 31, tinnitus in 27, distal extremity paresthesias in 22, joint pains in 13, low back pain in 5, and gait "ataxia" in 4. Symptoms resolved promptly upon lowering the intracranial pressure by lumbar puncture, and were probably directly caused by intracranial hypertension. Awareness of these "minor" symptoms of increased intracranial pressure can facilitate diagnosis and management. PMID- 2970602 TI - Digital subtraction angiography. PMID- 2970603 TI - Medical Practitioners Disciplinary Committee--medical certification. PMID- 2970604 TI - Low dose hepatitis B vaccination in children: benefit of low dose boosters. AB - Fifty eight children aged 5-12 years (mean 9.3 years) who had received three x 2 micrograms of Merck Sharp and Dohme (MSD) H-B-Vax intramuscularly at time 0, 1, 6 months were given a further 2 micrograms dose 12 months after dose 3 and tested for immune response two weeks later. Fifty seven of 58 children were positive for antibody to hepatitis B surface antigen (anti-HBs) when tested by radioimmunoassay. In 56 of the 57, the response was anamnestic in nature. The geometric mean titre of anti-HBs rose from 106 IU/L before, to 15,759 IU/L after the booster dose. The latter figure was greater than that obtained in 20 adults given three x 20 micrograms of the same vaccine and also greater than that reported in children given three x 10 micrograms of MSD H-B-Vax. This study demonstrates that 8 micrograms of H-B-Vax given as four doses is at least as immunogenic in children as 30 micrograms of the same vaccine given as recommended and that the low dose strategy is appropriate for this expensive vaccine. PMID- 2970605 TI - Accident consultations in one month in a semirural town. AB - General practice statistics for the month of July 1986 were analysed from a semirural town. There were 498 accident consultations and Maoris tended to be underrepresented. Thirty six percent of accidents were at work, 30% in the home and 12.6% in sports. Twenty three percent of the accidents involved no time off work and 32% had up to a week off. Thirty percent made a claim for earnings related compensation to ACC. Eight patients had prolonged periods off work. PMID- 2970606 TI - Atrial natriuretic factor and the atrium. PMID- 2970607 TI - Catapres-TTS: a two year posttrial followup. PMID- 2970608 TI - Compensated back injury in New Zealand. AB - Back injury is one of the commonest claims for accident compensation. A survey carried out over three months in 1984 identified 420 claimants having details of their accidents. Labourers, freezing workers, coal miners and railway workers were at high risk. Over half (54.7%) developed back pain whilst lifting and for almost two-thirds (63.6%) there was a sudden strain. Half had a previous history of back pain and the incidence rose to 70% for those aged 40-59 years. Eighty-two percent returned to work within four weeks. Most were treated by a general practitioner and physiotherapy was increasingly used as the period of disability lengthened. Rest and pain killers were assessed as the most popular forms of treatment. There was a need for education of the workforce in the ergonomics of lifting. PMID- 2970609 TI - Fenbufen: adverse reactions affecting the skin. PMID- 2970610 TI - Occupational overuse--important ACC decision. PMID- 2970611 TI - Evaluation of 99Tcm mercaptoacetyltriglycine for the detection and localization of gastrointestinal bleeding in an experimental animal model. AB - Technetium-99m mercaptoacetyltriglycine (99Tcm-MAG3) was evaluated for the detection and localization of the site of gastrointestinal bleeding in a sheep model. Radioactivity was detected in both the stomach and lower abdomen. However, 99Tcm-MAG3 is partially excreted by the liver and in the bile. The radioactivity in the bile moved to the small bowel. This movement of radioactivity in the lower abdomen can lead to a misinterpretation of the site of bleeding. Hence, 99Tcm MAG3 may not be an effective radiopharmaceutical for the localization of the site of bleeding in the gastrointestinal tract, particularly the lower abdomen. PMID- 2970612 TI - Therapy of the blast phase of chronic granulocytic leukemia with mithramycin and hydroxyurea. PMID- 2970613 TI - Identification of the protein product of the c-mos proto-oncogene in mouse testes. AB - The mouse c-mos proto-oncogene RNA is expressed primarily in mouse gonadal tissues and embryos. Until now, the c-mos protein has not been identified. Utilizing two different site-directed affinity purified anti-peptide antibodies, we have identified a 43 kDa c-mos protein in mouse testes and in germ cell preparations derived from testes. This 43 kDa testicular protein was found to be structurally related to a bacterially expressed c-mos protein by peptide mapping. Immunoblots of whole mouse sections were employed to establish that the c-mos protein is expressed primarily in the testes. PMID- 2970614 TI - Studies on stage-specific immunity against Taenia taeniaeformis metacestodes in mice. AB - The possible existence of stage-specific immune responses to Taenia taeniaeformis infection was investigated in C3H/He mice vaccinated with antigens prepared from either the oncosphere or metacestode stages. Mice were immunized twice, 2 weeks apart, with antigen in Freund's complete adjuvant. Two weeks after the second immunization they were challenged with 250 T. taeniaeformis eggs and killed day 0, 5, 10, 15, 20, 25, 30, 45 and 60 after infection. Gross examination of the livers revealed marked differences between oncosphere (TtO) and metacestode (TtM) vaccinated mice. Very few metacestodes were found in the first group but most of those that evaded the initial host attack developed like the cysts found in the control group. In contrast, many degenerating metacestodes were found in the TtM vaccinated group. In a subsequent experiment groups of mice were vaccinated with varying doses of either TtO or TtM to determine whether the qualitative differences observed above were due to antigen dose effects. However, varying antigen doses gave the same results. These data show that vaccination with oncospheres generates an immune response capable of killing invading larvae soon after infection whereas vaccination with TtM results in larvae being killed at a later stage, suggesting that there are stage-specific, host-protective antigens. PMID- 2970616 TI - Pain clinic #9. Physical exercise in the treatment of low back pain. Part I: A review. AB - Physical exercise has been advocated as therapeutic in the treatment of low back pain as far back as 1937, when Williams proposed his now famous flexion exercises. Its current widespread use invites two questions: 1) what evidence is there to support this claim, and 2) which exercises have been proven beneficial? In this first paper of a three-part series, the literature is examined for answers to these questions. Subsequent articles will present practical clinical regimens of both stretching and strengthening exercises. PMID- 2970615 TI - Analysis of antibody responses to Hymenolepis nana infection in mice by the enzyme-linked immunosorbent assay and immunoprecipitation. AB - Serum antibody responses in two strains of mice infected with embryonated eggs of Hymenolepis nana were analysed by the enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation (IP) using sodium deoxycholate (DOC)-solubilized antigens prepared from embryonated eggs (eggs), mouse-derived cysticercoids (cysts) and adult tapeworms with immature segments only (adults). Highly susceptible dd mice, which harbour mature tapeworms for a long period (greater than 70 days), produced high levels of antibodies to all three different stages of H. nana. BALB/c mice, almost all of which expel adult tapeworms by 30 days after infection, produced high levels of antibody against egg antigens only. The high antibody titres to cyst and adult antigens in dd mice did not lead to expulsion of the worms. However, worms are rejected early in BALB/c mice when there is little or no detectable serum antibody. The antibody responses to eggs seen in BALB/c mice which had long since shed their adult worms were probably due to ingestion of eggs from faeces of other infected mice. Antibodies to eggs were not detected in BALB/c mice which were initially inoculated with eggs (day 0) and then treated with praziquantel on day 6 after the tissue phase of infection only. The different antibody responses to egg antigens and the other two antigens (cyst and adult) in BALB/c mice suggest a difference in antigen specificity between eggs and both cysts and adults. A major antigen component with Mr 32,000 appears to be specific to the egg (or oncosphere) stage of H. nana. Antibody to this major component of eggs was absorbed only with intact eggs, but not with intact cysts nor adults with immature segments only, so that the antigen appears to be on the surface of the oncosphere. PMID- 2970617 TI - [Levels of erythropoiesis-regulating factors of the blood in patients with respiratory failure in chronic nonspecific lung diseases]. PMID- 2970618 TI - [Interrelation of different inflammatory processes occurring simultaneously in one organism]. PMID- 2970619 TI - Receptor-mediated uptake of acid alpha-glucosidase corrects lysosomal glycogen storage in cultured skeletal muscle. AB - Attempts at treatment of glycogenosis type II and other lysosomal storage disorders by enzyme replacement have been reported. Parenteral enzyme administration has been ineffectual. Treatment by bone marrow transplantation is currently under investigation. We have used cultured skeletal muscle cells from a patient with infantile glycogenosis type II to study fundamental aspects of enzyme replacement therapy. Efficient uptake of acid alpha-glucosidase was achieved by using the mannose-6-phosphate receptor on the cell surface as a target for an enzyme precursor with phosphorylated high-mannose types carbohydrate chains purified from human urine. We found that the enzyme was channeled to the lysosomes and converted to mature acid alpha-glucosidase. Glycogen storage was reversed. The results are discussed in relation to treatment of glycogenosis type II. PMID- 2970620 TI - Actuality in the treatment of unstable angina pectoris. AB - In an attempt to relieve ischaemic symptoms and to prevent progression to myocardial infarction, coronary angioplasty was attempted in 236 multivessel coronary heart disease patients with unstable angina, refractory to medical treatment including oral Ca2+ antagonists, beta blockers and nitroglycerin drugs. Unstable angina was defined as ischaemic chest pain at rest lasting for at least 20 min, accompanied by reversible ST-T changes. The initial angioplasty success rate was 87% (205/236 cases). Vessel occlusion necessitating urgent bypass surgery occurred in five patients (2.1%). There was evidence of myocardial infarction in eight patients (3.4%). There were seven deaths (2.9%) related to the procedure. 191 of the 205 successfully dilated patients were followed up for 14 months on the average. Late mortality occurred in 4.2% (8/191), late nonfatal infarction in 2.6%, 127 patients remained asymptomatic and 11 were considered to be disabling angina (New York Heart Association classification III or IV). Recurrent angina rate with progression in ischaemic disease (restenosis and native vessel stenosis) occurred in 30%. For this reason, repeated angioplasty and elective bypass surgery were performed in 48 and 14 cases, respectively. These results support the growing evidence that angioplasty as an emergency procedure in multivessel disease patients with unstable angina pectoris refractory to intensive medical treatment can restore coronary blood flow with an acceptable risk and a good initial and short-term success rate. PMID- 2970621 TI - Alterations of vasoconstrictor and sodium-regulating hormone systems in vascularly decompensated liver cirrhosis. AB - Plasma levels of atrial natriuretic peptide (ANP), aldosterone (PA), vasopressin (AVP), and the plasma renin activity (PRA) were examined in 15 vascularly decompensated patients suffering from liver cirrhosis, before and after administration of albumin and after a subsequent administration of furosemide. The initial ANP level was lower in 9 patients (group "A") and higher in 6 patients (group "B") than in healthy controls (Group "A": 19.5 +/- 3.0 fmol/ml; group "B": 36.7 +/- 3.9 fmol/ml; control: 25.8 +/- 2.4 fmol/ml). The initial PRA (4.4 +/- 1.0 ng AngI/ml/h) and AVP (8.5 +/- 1.5 pg/ml) activity in group "A" increased significantly compared to group "B" (PRA: 0.44 +/- 0.09; AVP: 4.1 +/- 0.5), indicating an intravascular volume depletion in group "A". Albumin infusion raised the urine and sodium excretion and the plasma concentration of ANP in group "A" but lowered in plasma levels of renin and vasopressin. The same parameters were not changed by albumin in group "B". Furosemide equally raised the urine flow rate and sodium excretion in both groups. Plasma ANP level depends on the intravascular volume, and the secondary change in its plasma concentration plays a considerable role in the retention of fluid and electrolytes in patients with cirrhosis. The increased intravascular volume in these patients depletes the fluid and electrolyte retention via the increase in ANP level. PMID- 2970622 TI - Mechanical circulatory assistance in the management of patients in coronary care unit. AB - Mechanical circulatory assistance was attempted in 73 patients at our coronary care unit from January 1, 1983, to January 1, 1987. Of these, 62 were men and 11 were women. The intra-aortic balloon pump (IABP) was successfully inserted by percutaneous femoral puncture in all cases. Indications for an IABP included: acute infarction and haemodynamic deterioration (23 cases), refractory congestive heart failure (2 cases), hypotension, ST-T changes and chest pain complicating coronarography or coronary angioplasty procedures (16 cases), unstable angina pectoris resistent to medical therapy, treated by coronary angioplasty (16 cases) and preoperative haemodynamic coverage (16 cases). The overall major complication rate of IABP was 1.5%. Our experience supports aggressive management of cardiogenic shock, i.e. early angioplasty or cardiac surgery, which considerably influences the functional status and the long-term survival rate (7/9 cases) over medical therapy combined with IABP (2 of the 14 patients are alive). The IABP was also effective in managing other high-risk patients when it was combined with some form of definitive cardioprotective mechanical correction, i.e. aneurysmectomy, valve replacement, coronary recanalization procedure. When the counterpulsation was used prophylactically, no interventional myocardial infarctions or deaths occurred; the survival rate was 27/32 patients for a 22 month average follow-up. It is suggested that the clinical results and late survival were improved by use of IABP in selected patients undergoing coronary angioplasty or open-heart surgery. PMID- 2970623 TI - The dual effect of thyroid hormones on contractile properties of rat myocardium. AB - This study was designed to investigate the changes in cardiac contractile properties induced by triiodothyronine (T3) administration in adult rats. Myofibrils and myosin were isolated from ventricular muscles from euthyroid and hyperthyroid animals and enzymatically and electrophoretically characterized. The time course of the isometric response, the force velocity curve, the force interval relation were studied in papillary muscles isolated from the right ventricles of euthyroid and hyperthyroid rats. T3 administration induced significant increases in Mg2+ activated myofibrillar ATPase activity (+11.4%) and in Ca2+ activated myosin ATPase activity (+20.1%). Significant increases in shortening velocity at low and zero loads (+20.4%) were found in papillary muscles from treated animals when compared with the control muscles. These variations in enzymatic activity and shortening velocity could be related to the increase in the amount of the fast isomyosin V1, as shown by pyrophosphate gel electrophoresis. The negative force-frequency relation at steady state, typical of rat cardiac preparations, was observed in treated and control animals; its slope was, however, halved in hyperthyroid papillary muscles when compared with control ones. In accordance with this finding, the potentiating effect of a prolonged diastolic interval was significantly reduced in hyperthyroid papillary muscles. In the frame of an interpretation of the force interval relation on the basis of the excitation contraction coupling processes, these latter observations might indicate an enhanced activity of the sarcoplasmic reticulum. We conclude that thyroid hormone administration has a dual effect on cardiac contractility, on one hand regulating the synthesis of the different isomyosin and, on the other hand, stimulating the activity of the sarcoplasmic reticulum. PMID- 2970624 TI - [Bone and muscle pain in dentists, attributed to their professional work]. PMID- 2970626 TI - Nucleotide sequence of the genome region encoding the structural proteins and the NS1 protein of the tick borne encephalitis virus. PMID- 2970625 TI - Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. AB - A lambda insertion type cDNA cloning vector, Lambda ZAP, has been constructed. In E. coli a phagemid, pBluescript SK(-), contained within the vector, can be excised by f1 or M13 helper phage. The excision process eliminates the need to subclone DNA inserts from the lambda phage into a plasmid by restriction digestion and ligation. This is possible because Lambda ZAP incorporates the signals for both initiation and termination of DNA synthesis from the f1 bacteriophage origin of replication (1). Six of 21 restriction sites in the excised pBluescript SK polylinker, contained within the NH2-portion of the lacZ gene, are unique in lambda ZAP. Coding sequences inserted into these restriction sites, in the appropriate reading frame, can be expressed from the lacZ promoter as fusion proteins. The features of this vector significantly increase the rate at which clones can be isolated and analyzed. The lambda ZAP vector was tested by the preparation of a chicken liver cDNA library and the isolation of actin clones by screening with oligonucleotide probes. Putative actin clones were excised from the lambda vector and identified by DNA sequencing. The ability of lambda ZAP to serve as a vector for the construction of cDNA expression libraries was determined by detecting fusion proteins from clones containing glucocerbrosidase cDNA's using rabbit IgG anti-glucocerbrosidase antibodies. PMID- 2970628 TI - [Contact dermatitis]. PMID- 2970627 TI - Working with Theresa. PMID- 2970629 TI - [First visit in pedodontics]. PMID- 2970630 TI - Evaluation of the patient with low back pain. PMID- 2970631 TI - Conservative management of low back pain. An evaluation of current methods. AB - The overwhelming majority of patients with low back pain recover, most of them within 6 to 12 weeks. Those who do not, however, represent a major challenge to physicians, who need to choose from among the many therapies for low back pain that have been advocated but in many cases not proven. For those with chronic low back pain, a functional restoration program and a multidisciplinary pain management program may prove effective. PMID- 2970632 TI - Diagnosis and treatment of spinal deformities. What to do for the patient with scoliosis. AB - Evaluation of scoliosis should include a careful medical history and a complete physical and neurologic assessment. Standing posteroanterior and lateral radiographs of the spine should be obtained and every effort made to arrive at an accurate diagnosis that pinpoints the cause of the spinal deformity. Special attention should be paid to painful spinal deformities in the pediatric population because they represent a potentially serious medical condition. Painful spinal deformities in adults usually respond to conservative therapy. Use of a spinal orthosis may help control pain in an adult and may prevent progression of a spinal deformity in a child or adolescent. Regardless of the patient's age, however, progressive or painful scoliosis may necessitate surgical treatment. PMID- 2970633 TI - Determination of fiber types of chicken skeletal muscles based on reaction for actomyosin, calcium+2, magnesium+2-dependent adenosine triphosphatase. AB - Muscle fiber subtypes, determined with the actomyosin Ca+2,Mg+2-adenosine triphosphatase (ATPase) reaction in chicken anterior latissimus dorsi and posterior latissimus dorsi muscles, were demonstrated only after acid or alkaline preincubation followed by a 60-min enzyme incubation. In contrast, subtypes were demonstrated in the sartorius muscle either with or without preincubation. A single-step procedure was therefore possible with this muscle. The results were generally similar to those obtained previously with the mycosin Ca+2-ATPase procedure. Both methods revealed corresponding muscle fiber subtypes, with the exceptions noted below. The actomyosin Ca+2,Mg+2-ATPase procedure, following preincubation at pH 9.4 and 10.3, resulted in a similar reaction intensity in all fiber types. With the myosin Ca+2-ATPase procedure, the IRA (slow) type in anterior latissimus dorsi and sartorius muscles and the I (slow), IIR (fast oxidative-glycolytic), and IIW (fast glycolytic) types in posterior latissimus dorsi muscle had a higher reaction intensity following preincubation at pH 9.4 than at pH 10.3. Fiber Types IIR and IIW in sartorius muscle were easily distinguished with the actomyosin Ca+2,Mg+2-ATPase procedure. PMID- 2970634 TI - CD3-associated heterodimeric polypeptides on suppressor hybridomas define biologically active inhibitory cells. AB - We have investigated the relationship between CD3 expression and the suppressor T cell function. We have isolated stable clonal cell lines of the F12.23 suppressor T-cell hybridoma that are either CD3+ or CD3-. These lines were subjected to functional assays including inhibition of in vivo hapten-specific delayed-type hypersensitivity responses, in vitro hapten-specific interleukin 2 responses, as well as hapten-specific cytotoxic T-lymphocyte assays. In all assays, the functional suppressor phenotype absolutely correlated with CD3 surface expression. Furthermore, we have immunoprecipitated heterodimeric proteins that share molecular features with some receptor polypeptides previously described. CD3 polypeptides found on the surface of suppressor T cells are phosphorylated after phorbol ester stimulation. Collectively these studies unambiguously define the suppressive supernatant function as a product of CD3+ receptor-bearing T cells. PMID- 2970635 TI - Molecular characterization of T-cell antigen receptor expression by subsets of CD4- CD8- murine thymocytes. AB - Precursors of all T-lineage cells are found in a population of thymocytes that lack the CD4 and CD8 surface glycoproteins. These "double-negative" thymocytes are markedly heterogeneous in their expression of other surface markers and include cells at various stages of development. In this study, CD4- CD8- adult murine thymocytes were separated into subsets based on the expression of the "heat stable antigen" (HSA) and of Ly 1 (CD5). The sorted subsets were analyzed directly (without prior expansion in culture) for T-cell antigen receptor (TcR) gene rearrangement and mRNA expression and for TcR and CD3 cell-surface protein expression. Very little surface CD3 or TcR expression was detected on the major HSA+ Ly 1low subset. However, the HSA+ Ly 1high, HSA- Ly 1high, and HSA- Ly 1low subsets all contained cells with surface expression of CD3 and TcR. In contrast to previous studies, we found no subset that exclusively expressed either the alpha beta or gamma delta heterodimer, although the ratio of alpha beta+ to gamma delta+ varied widely. Two of these three subsets (HSA- Ly 1low and HSA- Ly 1high) showed very high usage of V beta 8 gene products in the alpha beta heterodimer, but nevertheless included approximately equal to 15% non-V beta 8 alpha beta forms. All CD4- CD8- subsets were found to have extensively rearranged their TcR gamma genes and to express gamma mRNA. Expression of a high ratio of mature [1.3 kilobases (kb)] to truncated (1.0 kb) beta message and presence of alpha message was largely restricted to subsets with TcR alpha beta surface expression. PMID- 2970636 TI - Reexpression of a developmentally regulated antigen in Down syndrome and Alzheimer disease. AB - ALZ-50 is a monoclonal antibody that recognizes a protein of apparent molecular mass 68 kilodaltons (A68). The protein is present in the brains of patients with Alzheimer disease but is not detectable in normal adult brain tissue. We now report that ALZ-50-reactive neurons are found in normal fetal and neonatal human brain and in brain tissue from neonatal individuals with Down syndrome. Reactive neurons decrease sharply in number after age 2 and reappear in older individuals with Down syndrome and in patients with Alzheimer disease. PMID- 2970637 TI - Purification and partial characterization of a receptor protein for mouse interferon gamma. AB - A receptor protein for mouse interferon gamma has been purified from solubilized plasma membranes of the mouse monomyelocytic cell line WEHI-3. Sequential wheat germ agglutinin and ligand affinity chromatography of membranes extracted with octyl beta-D-glucopyranoside resulted in at least a 680-fold purification of the receptor, as measured by precipitating it in association with liposomes composed of phosphatidylcholine. The purified receptor bound 125I-labeled recombinant mouse interferon gamma (rMuIFN-gamma) with a Kd of 10 nM, a value comparable to that obtained with isolated membranes (3.5 nM). PAGE analysis of radiolabeled (with either 35S or 125I) receptor preparations consistently revealed a major band of 95 kDa. This species was degraded with time to smaller fragments, principally one of 60 +/- 5 kDa. Treatment with peptide:N-glycosidase F reduced the apparent molecular masses of the proteins in the 95- and 60-kDa regions by 15 20 kDa each. GR-20, a monoclonal antibody against the receptor, completely inhibited specific binding of 125I-labeled rMuIFN-gamma to WEHI-3 cells, blocked the induction of priming by rMuIFN-gamma of macrophage-mediated tumor cell killing, removed binding activity for 125I-labeled rMuIFN-gamma from solubilized membranes, and immunoprecipitated a single 95-kDa protein from the extract of surface labeled (125I) WEHI-3 cells. Cross-linking of 125I-labeled rMuIFN-gamma to its receptor yielded a complex of 125 +/- 5 kDa, consistent with the binding of the dimeric form of mouse interferon gamma (32 kDa) to a membrane protein of 95 kDa. These data suggest that the receptor for mouse interferon gamma (or a ligand-binding subunit thereof) is a glycoprotein of 95 kDa. PMID- 2970638 TI - Kinesin ATPase: rate-limiting ADP release. AB - The ATPase rate of kinesin isolated from bovine brain by the method of S.A. Kuznetsov and V.I. Gelfand [(1986) Proc. Natl. Acad. Sci. USA 83, 8530-8534)] is stimulated 1000-fold by interaction with tubulin (turnover rate per 120-kDa peptide increases from approximately equal to 0.009 sec-1 to 9 sec-1). The tubulin-stimulated reaction exhibits no extra incorporation of water-derived oxygens over a wide range of ATP and tubulin concentrations, indicating that Pi release is faster than the reversal of hydrolysis. ADP release, however, is slow for the basal reaction and its release is rate limiting as indicated by the very tight ADP binding (Ki less than 5 nM), the retention of a stoichiometric level of bound ADP through ion-exchange chromatography and dialysis, and the reversible labeling of a bound ADP by [14C]ATP at the steady-state ATPase rate as shown by centrifuge gel filtration and inaccessibility to pyruvate kinase. Tubulin accelerates the release of the bound ADP consistent with its activation of the net ATPase reaction. The detailed kinetics of ADP release in the presence of tubulin are biphasic indicating apparent heterogeneity with a fraction of the kinesin active sites being unaffected by tubulin. PMID- 2970639 TI - Increased mRNA expression of a laminin-binding protein in human colon carcinoma: complete sequence of a full-length cDNA encoding the protein. AB - Reliable markers to distinguish human colon carcinoma from normal colonic epithelium are needed particularly for poorly differentiated tumors where no useful marker is currently available. To search for markers we constructed cDNA libraries from human colon carcinoma cell lines and screened for clones that hybridize to a greater degree with mRNAs of colon carcinomas than with their normal counterparts. Here we report one such cDNA clone that hybridizes with a 1.2-kilobase (kb) mRNA, the level of which is approximately equal to 9-fold greater in colon carcinoma than in adjacent normal colonic epithelium. Blot hybridization of total RNA from a variety of human colon carcinoma cell lines shows that the level of this 1.2-kb mRNA in poorly differentiated colon carcinomas is as high as or higher than that in well-differentiated carcinomas. Molecular cloning and complete sequencing of cDNA corresponding to the full length open reading frame of this 1.2-kb mRNA unexpectedly show it to contain all the partial cDNA sequence encoding 135 amino acid residues previously reported for a human laminin receptor. The deduced amino acid sequence suggests that this putative laminin-binding protein from human colon carcinomas consists of 295 amino acid residues with interesting features. Containing only two cysteine residues, the protein does not have consensus sequences for asparagine-linked glycosylation, amphipathic alpha-helix, or the N-terminal leader signal sequences for entry into endoplasmic reticulum, although hydrophobic segments for potential membrane associations exist. There is an unusual C-terminal 70-amino acid segment, which is trypsin-resistant (no lysine or arginine) and highly negatively charged (13 aspartic plus glutamic residues). Within this segment are five repeats of (Asp/Glu)-Trp-(Ser/Thr); two of these are nearly tandem repeats of Thr Glu-Asp-Trp-Ser-Ala-Xaa-Pro. PMID- 2970641 TI - Blockade of the interleukin-2 receptor by anti-Tac antibody inhibits the generation of antigen-nonspecific suppressor T cells in vitro. AB - The role of interleukin 2 (IL-2) in the activation of suppressor T cells was investigated by using the monoclonal antibody anti-Tac, which blocks the binding of IL-2 to the 55-kDa peptide of the high-affinity IL-2 receptor. Anti-Tac was added to an antigen-nonspecific suppressor system in which Con A-induced suppressor T cells were generated during a preculture period, and their effects on immunoglobulin production were assessed in second, indicator cultures containing pokeweed mitogen and peripheral blood mononuclear cells. Anti-Tac added during the preculture period inhibited Con A-induced suppressor T-cell generation. Cells activated by a short (2-day) preculture period to become effectors of suppression were primarily of the Tac-positive, T8 (CD8)-positive phenotype. Tac-positive, T8-negative T cells might also contribute to the suppressor activity. Our studies indicate that anti-Tac, by producing a functional blockade of human high-affinity IL-2 receptors, inhibits the generation of antigen-nonspecific suppressor T cells. PMID- 2970640 TI - The 289-amino acid E1A protein of adenovirus binds zinc in a region that is important for trans-activation. AB - The E1A gene of adenovirus type 5 encodes two major proteins of 289 and 243 amino acid residues, which are identical except that the larger protein has an internal stretch of 46 amino acids required for efficient trans-activation of early viral promoters. This domain contains a consensus zinc finger motif (Cys-Xaa2-Cys-Xaa13 Cys-Xaa2-Cys) in which the cysteine residues serve as postulated ligands. Atomic absorption spectrophotometry applied to bacterially expressed E1A proteins revealed that the 289-amino acid protein binds one zinc ion, whereas the 243 amino acid protein binds no zinc. Replacing individual cysteine residues of the finger with other amino acids destroyed the trans-activating ability of the 289 amino acid protein, even when structurally or functionally conserved amino acids were substituted. These results strongly suggest that the zinc finger of the 46 amino acid domain is intimately linked to the ability of the large E1A protein to stimulate transcription of E1A-inducible promoters. Furthermore, zinc binding to one of the mutant finger proteins suggests either that only a precise finger structure formed by the tetrahedral coordination of zinc to the four consensus ligands is required for trans-activation or, possibly, that one of several neighboring histidine residues in various combinations with three of the consensus cysteine residues normally coordinates zinc. How the zinc finger in E1A might interact with DNA or protein to bring about trans-activation is discussed. PMID- 2970642 TI - Isolation and characterization of cDNAs coding for the beta subunit of the high affinity receptor for immunoglobulin E. AB - Among receptors that bind the Fc region of immunoglobulins ("Fc receptors"), only the one with high affinity for immunoglobulin E (IgE) is known to consist of more than a single polypeptide. In addition to the IgE-binding alpha chain, the receptor contains a single beta chain and two, disulfide-linked, gamma chains. From a cDNA library of a rat mucosal mast cell tumor, from which we recently cloned cDNAs coding for the alpha chain, we have now isolated cDNAs coding for the beta subunit. In vitro transcription-translation of the cDNA directed the synthesis of a polypeptide reactive with two distinctive anti-beta monoclonal antibodies and whose molecular weight was identical to that of authentic beta chains. Polyclonal antibodies to beta peptides expressed in Escherichia coli reacted with intact receptors and isolated beta chains. The gene encodes a protein of 243 residues with no leader sequence. A hydropathicity plot suggests that the polypeptide crosses the plasma membrane four times. The epitope recognized by one of the monoclonal antibodies was localized to the NH2 terminus; that by the other was localized to the COOH terminus. Since those antibodies react with membranes and not with intact cells, we suggest that both ends of the beta subunit are cytoplasmic. RNA transfer blots at high stringency failed to reveal mRNA for beta chains in a variety of cells (in particular, monocytes) that do not contain the high-affinity receptor for IgE. PMID- 2970643 TI - Role of the Escherichia coli DnaK and DnaJ heat shock proteins in the initiation of bacteriophage lambda DNA replication. AB - We examined the role of two Escherichia coli heat shock proteins, the dnaK and dnaJ gene products, during the initiation of lambda dv DNA replication in vitro. Using 14C-labeled lambda P protein we showed that the DnaK and DnaJ heat shock proteins function together to release lambda P protein from the preprimosomal complex consisting of lambda origin of replication-lambda O-lambda P-DnaB protein. Hydrolysis of ATP, catalyzed presumably by DnaK, is required during this reaction. Substitution of DnaK protein with that of the mutant DnaK756 protein blocks lambda P release. After DnaK and DnaJ action, the preprimosomal complex, isolated on Sepharose 4B, can support lambda dv DNA replication without any additional prepriming proteins. Using DnaK-affinity chromatography we showed that both lambda O and lambda P proteins bind to DnaK protein. The lambda P protein interacts with DnaK protein in a salt-resistant, hydrophobic manner, and ATP hydrolysis is necessary to elute at least part of lambda P protein from the DnaK affinity column. The proposed mechanism of action of the prokaryotic DnaK and DnaJ heat shock proteins agrees with the hypothesis that Hsp70, the DnaK analogue of eukaryotes, uses ATP to disrupt hydrophobic aggregates [Pelham, H. R. B. (1986) Cell 46, 959-961]. PMID- 2970645 TI - How labels affect public attitudes to people with handicaps. PMID- 2970644 TI - IgE production by normal human lymphocytes is induced by interleukin 4 and suppressed by interferons gamma and alpha and prostaglandin E2. AB - The effect of human recombinant interleukin 4 (IL-4) on antibody production by normal peripheral blood mononuclear cells enriched for B cells was investigated. IL-4 preferentially induced IgE synthesis in vitro. In addition, a low induction of IgG production was observed, whereas IL-4 had no effect on IgA and IgM synthesis. The IL-4-induced IgE production by B cells required T cells and monocytes but was specifically inhibited by an anti-IL-4 antiserum indicating that, although IL-4 acts indirectly, it is responsible for the induction of IgE synthesis. IL-4-induced IgE production was blocked in a dose-dependent way by interferon gamma (IFN-gamma), interferon alpha (IFN-alpha), and prostaglandin E2. IFN-gamma also inhibited IL-4-induced IgG production. These inhibitory effects of IFN-gamma and IFN-alpha on IgE production cannot be attributed to toxic effects since IFN-alpha induced IgM production in the presence of IL-4, whereas IFN-gamma was ineffective in inhibiting IgG production induced by IL-2. IFN-gamma, IFN alpha, and prostaglandin E2 also inhibited IL-4-induced expression of the low affinity receptor for the Fc portion of IgE (CD23) on B cells, indicating that there is an association between CD23 expression and IL-4-induced IgE production. This theory was supported by the finding that IL-4-induced IgE production was inhibited by F(ab')2 fragments of an anti-CD23 monoclonal antibody. PMID- 2970646 TI - Intracellular transport of the mannose-6-phosphate receptor. PMID- 2970647 TI - Phosphomannosyl receptor binding is localized in vesicles separated by free-flow electrophoresis from rough endoplasmic reticulum preparations of rat liver. PMID- 2970648 TI - [Interference with the urine analysis of etodolic acid]. PMID- 2970649 TI - 8-OH-DPAT elicits feeding and not chewing: evidence from liquid diet studies and a diet choice test. AB - There have been recent claims that the 5-HT1A agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) elicits chewing and eating of solid but not liquid foods. Therefore, the effects of 8-OH-DPAT and another 5-HT1A agonist gepirone on the consumption of a liquid chow diet, by free feeding male rats, were examined. Both drugs produced a dose-dependent increase in the consumption of liquid diet during a 2 h test. The doses of 8-OH-DPAT and gepirone which increased liquid diet intake in this study were in the same range as those which were found previously to increase food pellet consumption by free feeding rats. The effects of 8-OH-DPAT were also examined in a feeding choice test in which free feeding animals were allowed to choose between food pellets and a liquid chow diet. In this test, 8-OH-DPAT significantly increased total food intake (liquid plus pellet) but had no significant effect on the consumption of either liquid or pellet diets when analysed separately. Thus, there were large individual differences in diet choice after 8-OH-DPAT injection. However, rats did not consistently choose to eat food pellets rather than the liquid diet, as would be predicted if the drug elicited chewing rather than eating. These results provide strong evidence that 8-OH-DPAT elicits a behaviourally specific hyperphagia and not chewing or gnawing. PMID- 2970650 TI - [Anxiety in disabled persons in the light of Antoni Kepinski's concept]. PMID- 2970651 TI - [Contributions to the biochemical study of saliva in periodontal disease and gingivitis in humans]. PMID- 2970652 TI - [Periodontium alterations in hypophyseal failure (Sheehan disease)]. PMID- 2970653 TI - [Aspects of occlusal and physionomic reorganization in the immediate total prosthesis]. PMID- 2970654 TI - [Cytophysiologic behavior of buccal mucosa in contact with the acrylic prosthesis, with and without styren additives]. PMID- 2970655 TI - [Longitudinal study on the age and eruption sequences of permanent teeth in the second stage of mixed dentition. Orthodontic implications]. PMID- 2970656 TI - [Iatrogenic aspects in orthodontics]. PMID- 2970657 TI - [Post-surgical orthodontic treatment in labio-vello-palatine clefts with the Frankel III device]. PMID- 2970658 TI - [Comparative data concerning the provision of assistance on request between 1979 and 1984 in the no. 1 Polyclinic Dispensary from Baia Mare]. PMID- 2970659 TI - [Value of duplication of total prostheses]. PMID- 2970660 TI - [A device for measuring the thickness of the occluding surfaces of cast crowns]. PMID- 2970661 TI - [Backache in a hospital environment: epidemiologic aspects and the role of various risk factors]. AB - A cross-sectional study was carried out in 1985 among 5941 hospital workers from the Regional and University Hospital Center (CHRU) of Strasbourg on the occasion of the occupational medical examinations. The purpose was to determine the importance of low back pain among hospital staff and to describe the relationship between individual and occupational characteristics and the onset of these symptoms. Our results confirm the risk-factors determined by other investigators: age, sex, length of service, heavy work assignments (nursing aides, stretcherbearers, nurses, etc.) but also underline the role of obesity and previous attacks of low back pain, and invalidate the part musculoskeletal abnormalities play in this pathology. PMID- 2970662 TI - [Defects of the atrioventricular septum with a small left ventricle]. PMID- 2970663 TI - [Ebstein anomaly in a 72-year-old woman]. PMID- 2970664 TI - [Familial idiopathic enlargement of the right atrium]. PMID- 2970665 TI - [Angioplasty of peripheral arteries]. PMID- 2970666 TI - [Correct sitting posture--an important element in the therapy and prevention of chronic back complaints. Ergonomic requirements for correct furniture for sitting]. PMID- 2970667 TI - Zinc and copper in plasma and erythrocytes of Down's syndrome children. AB - The concentration of zinc and copper in the plasma and the erythrocytes of 24 children with Down's syndrome was measured and compared with the values in a control group of normal children. Zinc and copper were determined in this biologic material by flame atomic absorption spectrophotometry. A significant (p less than 0.001) decrease of the plasma zinc content well as an increase of copper (p less than 0.024) and zinc (p less than 0.001) in the erythrocytes of Down's syndrome patients were found. The possible mechanisms of these changes are discussed. PMID- 2970668 TI - Abnormal helper-inducer/suppressor-inducer T-cell subset distribution and T-cell activation status are common to all types of chronic synovitis. AB - We have previously shown that rheumatoid synovial T cells are virtually all helper-inducer (CD4+4B4+UCHL1+) rather than suppressor-inducer (CD4+2H4+) cells. CD8 cells were also largely 4B4+. In addition, the majority of T cells were HLA DR+. To investigate whether these findings were specific for rheumatoid disease, we studied the prevalence of these markers in a variety of chronic inflammatory arthropathies such as ankylosing spondylitis, Reiter's syndrome, and psoriatic arthritis. Again, almost 90% of the T cells were 4B4+UCHL1+ and only 11% were 2H4+; 50% expressed the HLA DR antigen. Thus this phenotypic distribution represents a final common pathway of chronic synovitis and may help to explain the immunopathology of the lesion. PMID- 2970669 TI - [Advances in studying the brain peptides]. PMID- 2970670 TI - Isotypic exclusion of gamma delta T cell receptors in transgenic mice bearing a rearranged beta-chain gene. AB - The rearrangement of T cell antigen receptor beta- and gamma-chain gene segments was studied in transgenic mice that bear a functional beta-chain gene. Virtually all CD3-positive T cells derived from transgenic mice express beta chains containing the transgene-encoded V beta 8.2 variable region on their surfaces and do not express endogenous beta-chain variable regions. Expression of endogenous V beta genes is inhibited at the level of somatic recombination during thymic ontogeny. Furthermore, rearrangements of the TCR gamma-chain genes are also markedly inhibited in these transgenic animals. Hence expression of the TCR beta transgene has led to allelic exclusion of alpha beta receptors and isotypic exclusion of gamma delta T cell receptors. PMID- 2970671 TI - The human laminin receptor is a member of the integrin family of cell adhesion receptors. AB - A receptor for the adhesive basement membrane protein, laminin, was isolated from human glioblastoma cells by affinity chromatography on laminin. This receptor has a heterodimeric structure similar to that of receptors for other extracellular matrix proteins such as fibronectin and vitronectin. Incorporation of the laminin receptor into liposomal membranes makes it possible for liposomes to attach to surfaces coated with laminin. The receptor liposomes also attached to some extent to surfaces coated with fibronectin, but not with other matrix proteins. These properties identify the laminin receptor as a member of the integrin family of cell adhesion receptors. PMID- 2970672 TI - Assembly of functional U1 and U2 human-amphibian hybrid snRNPs in Xenopus laevis oocytes. AB - Oligonucleotides complementary to regions of U1 and U2 small nuclear RNAs (snRNAs), when injected into Xenopus laevis oocytes, rapidly induced the specific degradation of U1 and U2 snRNAs, respectively, and then themselves were degraded. After such treatment, splicing of simian virus 40 (SV40) late pre-mRNA transcribed from microinjected viral DNA was blocked in oocytes. If before introduction of SV40 DNA into oocytes HeLa cell U1 or U2 snRNAs were injected and allowed to assemble into small nuclear ribonucleoprotein particle (snRNP)-like complexes, SV40 late RNA was as efficiently spliced as in oocytes that did not receive U1 or U2 oligonucleotides. This demonstrates that oocytes can form fully functional hybrid U1 and U2 snRNPs consisting of human snRNA and amphibian proteins. PMID- 2970673 TI - Selection of variable-joining region combinations in the alpha chain of the T cell receptor. AB - Most T lymphocytes express an antigen-specific receptor composed of two subunits, alpha and beta, each of which can exhibit structural variability. A complex selection process operates on T cells during development in the thymus such that cells expressing only particular alpha beta-receptors migrate to the periphery. The alpha-chain repertoire was dissected at different stages of the selection process by using the polymerase chain reaction (PCR) technique to amplify only those transcripts of a particular variable region gene (V58). Sequences from these V58 cDNAs reveal the predominant expression of four joining (J) segments by T cells in the adult thymus, suggesting that molecular or cellular processes select particular V alpha J alpha combinations during development. T cells expressing one of these V58J alpha chains appear to have been negatively selected at a later stage, since these transcripts were present in the spleen at approximately one-tenth the level in the thymus. Results also indicate that residues present at the V alpha J alpha junction may be important in an early selection process. PMID- 2970674 TI - [Health status of schoolchildren from general-education boarding schools and of inmates of children's homes]. PMID- 2970675 TI - [Comparative characteristics of morbidity among young children in complete and incomplete families]. PMID- 2970676 TI - [The organization of ultrasonic diagnosis]. PMID- 2970678 TI - [Medical workers--active fighters in establishing Soviet rule in Azerbaijan]. PMID- 2970677 TI - [Organization of scientific research in medical schools]. PMID- 2970679 TI - [A organizer of higher medical education, scientist and Medical Society founder S.S. Salazkin]. PMID- 2970680 TI - Altered form of subunit 6 of mitochondrial ATP synthase complex in oligomycin resistant mutants of Chinese hamster ovary cells. AB - Using an antiserum generated against a synthetic peptide predicted from the DNA sequence of the ATPase 6 gene of the mitochondrial DNA, we demonstrate that mitochondria from two oligomycin-resistant Chinese hamster ovary cell lines with a defined mutation in the ATPase 6 gene synthesize an altered ATPase 6 gene product. This altered gene product migrates in sodium dodecyl sulfate polyacrylamide gels as if it has a molecular mass that is larger by 1000 daltons than the wild-type ATPase 6 gene product. We also demonstrate that mitochondria from four other independently isolated oligomycin-resistant Chinese hamster ovary mutant cell lines contain a similar altered ATPase 6 gene product. These results suggest that all six oligomycin-resistant cell lines have a similar mutation in the ATPase 6 gene of the mitochondrial DNA that encodes subunit 6 of the ATP synthase complex. PMID- 2970681 TI - Genetic mapping in chromosome 21 and its implications for Down's syndrome and other diseases. PMID- 2970682 TI - Value of diagnostic laparoscopy in young women with possible appendicitis. AB - Removal of a normal appendix because of suspected appendicitis occurs most frequently in women of reproductive age. We investigated the value of laparoscopy in the diagnostic evaluation for possible appendicitis in women of childbearing age. Fifty-one women were entered in a management protocol, which included diagnostic laparoscopy in instances with atypical features. Twenty patients underwent immediate appendectomy because of history and physical findings classical for appendicitis; 31 women with atypical history and physical findings underwent an initial diagnostic laparoscopy. In the group of patients who underwent immediate appendectomy because of classical presentation, five of the appendices removed were normal. At laparoscopy, appendicitis was diagnosed in five patients, another disease in 15 and no diagnostic abnormality was found in six. Five patients had a normal appendix removed, two because of questionable inflammation and three because of nonvisualization of the appendix. There were no false-negative examination results. Twenty-one patients required no further operative intervention after laparoscopy. The negative appendectomy rate remained unchanged (ten negative appendectomies of 30 performed). Definitive diagnoses of eight instances of pelvic inflammatory disease, six of ruptured ovarian cysts and one instance of ileitis of the small intestine were made earlier than would have been possible without diagnostic laparoscopy in this setting. Diagnostic laparoscopy permits earlier definitive diagnosis and prompt institution of appropriate therapy for disease of the female reproductive tract that simulates appendicitis. Caution is advised, however, when diagnostic laparoscopy is applied more frequently than right lower quadrant exploration in the management of probable appendicitis. The improvement in diagnostic accuracy may be offset by an increased number of negative appendectomies resulting from nonvisualization and false-positive inflammation. PMID- 2970683 TI - Early experience with laparoscopic appendectomy in women. AB - Experience and the surgical technique of laparoscopic appendectomy in 70 female patients over a period of more than three years is described. Three women were pregnant. Complications occurred in one case. PMID- 2970684 TI - Laparoscopic ovarian electrocautery in patients with polycystic ovarian disease resistant to clomiphene citrate. AB - Eleven patients with polycystic ovarian disease resistant to clomiphene citrate were treated by ovarian electrocautery. Nine patients ovulated after electrocautery, and five became pregnant (four-following additional treatment with clomiphene citrate). Testosterone and androstenedione levels were reduced, but androstenedione levels returned to pretreatment values at 6 months. PMID- 2970685 TI - An experimental model for the combined medical and surgical treatment of gallstones. AB - New approaches to the treatment of gallstone disease have been introduced in recent years. In addition to bile acid dissolution, a chemical solvent (methyl tertiary-butyl ether) has been instilled directly into the gallbladder by percutaneous transhepatic techniques. Shock wave fragmentation of stones, induced by extracorporeal techniques or contact catheter probe placement, have also been successfully employed. Recurrence of gallstones, however, can be anticipated in most of these patients with an intact gallbladder and continued hepatic production of lithogenic bile. In order to explore these new procedures and to attempt to find a solution to the problem of stone recurrence, an animal model has been developed. The essential components of this model consist of an endoscopic (laparoscopic) cholecystopexy and, at a later date, percutaneous re entry into the gallbladder by dilation of the cholecysto-cutaneous tract and the use of electrohydraulic and ultrasonic lithotripsy and methyl tertiary-butyl ether to achieve stone dissolution. PMID- 2970688 TI - [Safe dental treatment and AIDS]. PMID- 2970687 TI - Effect of photodynamic therapy on blood flow in normal and tumor vessels. AB - The aim of this series of experiments was to determine the dynamic blood flow changes that occur in normal and neoplastic tissues during photodynamic therapy. Mice bearing SMT-F tumors and rats with transplanted chondrosarcomas were injected with graded doses of dihematoporphyrin ether. Studies of changes in single-vessel and whole-tumor blood flow were carried out with 630 nm light activation. A helium neon laser Doppler velocimeter was used to stimulate dihematoporphyrin ether, as well as to measure changes in flow velocity in both single-vessel and whole-tumor models. There was a reduction of flow velocity in all vessels and tumors in animals injected with 1 to 40 mg/kg dihematoporphyrin ether intraperitoneally. The extent of flow reduction was related to drug dose administered. Decreases in blood flow began within 10 seconds of light stimulation and were maximal within 5 minutes. Both normal and tumor vessels responded similarly. We conclude that photodynamic therapy leads to significant microcirculatory changes that may be pertinent to the mechanism of tumor necrosis. PMID- 2970686 TI - Clearance of cryoglobulins in man. AB - From the foregoing it is evident that the defective clearance of immune complexes may contribute to tissue damage seen in patients with cryoglobulins. Hypocomplementaemia, decreased erythrocyte CR1, and the nature of the immune complexes may all contribute to decreased binding of immune complexes to erythrocytes with the potential consequence of increased availability for deposition and decreased processing of immune complexes. In addition, in type II cryoglobulinaemia the nature of the immune complex namely IgM RF/IgG complexes, has been shown in some circumstances to fix C3 and C4 inefficiently in spite of detectable fluid phase complement activation. The poor C3 fixation results not only in decreased transport by erythrocytes to the RES but also in inefficient removal of immune complexes by phagocytic cells, since the Fc receptor on these cells would be acting alone without the synergy of occupied complement receptors. Persistence of immune complexes in tissues in these circumstances has two potential consequences. First, the multivalency of IgM may contribute to enlargement of these immune complexes in situ by successive trapping of antigen and antibody complexes; this process may be aided by local factors such as low temperature at the peripheries and increased protein concentration in glomerular capillary loops, which favour "cryo" precipitation. The higher avidity of RF for IgG that is surface bound rather than monomeric would also favor immune complex formation in these circumstances. Second, although there is no opsonization of the immune complex complement activation results in bystander fixation of C4 and C3 to the surrounding tissues resulting in tissue injury. PMID- 2970689 TI - Great variation in the response of tissue plasminogen activator activity, plasminogen activator inhibition and plasmin inhibition to endotoxin, aspirin and endotoxin after administration of aspirin. AB - The effect of endotoxin, aspirin and endotoxin after administration of aspirin on the tissue plasminogen activator activity (PAA), plasminogen activator inhibition (PAI) and plasmin inhibition (PI) was studied in the rat. PAA, PAI and PI were determined in key organs (brain, heart, lungs, kidneys, liver and aorta) spectrophotometrically by procedures involving hydrolysis of the chromogenic substrate S-2251. Aspirin at three different doses had not any significant effect on tissue PAA; PAI and PI were affected in several organs. Four hours after a sustained infusion of endotoxin PAA was found to be increased in brain, kidneys and aorta, decreased in heart and lungs, while in liver the PAA was unchanged compared to controls. Changes in PAI and PI showed also a tissue variation. In endotoxin-infused rats pretreated with aspirin the PAA changes induced by endotoxin were prevented or modified; PAI and PI were affected in most organs studied. However, this effect of aspirin was varying and depending on the tissue, the parameter studied and the dose of aspirin. In some organs, as the heart and lungs, changes in PAI or PI were noticed, while neither aspirin nor endotoxin separately induced such changes in these organs. Also, a differential response of PAI and PI to the same stimulus in the same tissue was a noteworthy finding. The results of the present study show that the response of PAA, PAI and PI to aspirin depends on the tissue, the physiological or pathophysiological condition of the tissue and the dose of the aspirin. PMID- 2970691 TI - Comparative study of different laser systems with special regard to angioplasty. AB - Laser angioplasty has found increasing interest in the treatment of cardiovascular diseases. First clinical applications of laser angioplasty in atherosclerotic stenoses or occlusions of peripheral and coronary vessels primarily showed a high success rate, but later were followed by significant complications as aneurysm formation, late perforations and restenoses. Most of these complications were due to a thermal damage of surrounding wall structures. To avoid these complications an ideal laser system should only minimally influence surrounding tissue layers and should offer a well predictable penetration and ablation including also severely calcified material. In conventional continuous wave laser systems photo energy is mainly changed into thermal energy. Mode of application, local cooling and the use of special laser probes can reduce the thermal side effects, but nevertheless histologic examinations revealed thermal injuries of various degree to surrounding structures. Furthermore these laser systems mostly faile to ablate or penetrate calcified plaques and only pulsed Nd:YAG-lasers with high frequencies seem ot offer some improvement. Eximer lasers with their wave lengths of 193, 248 and 308 nanometers also work in a high frequent pulsed mode. They offer a very low penetration and mainly show a so called photoablative effect with only minimal thermal side effects. Within a distance of only 50 micrometers temperature raises in the surrounding tissue up to 5 degrees C and in a distance of 1 mm, up to 1.5 degrees C. Eximer lasers are moreover able to penetrate calcified material. A transluminal application is up till now limited by a lack of flexible fiber systems, which can deliver the high energy pulses. Another point of discussion are the mutagenity and cancerogenity of ultraviolet light.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970690 TI - Platelet and clotting activities after cold stress in diabetic patients. AB - Platelet and clotting abnormalities have been described in diabetes, but little is known about their relationship to daily stresses. In order to evaluate whether states of abnormal carbohydrate metabolism modify the hemostatic response to stress, 12 subjects with type I diabetes mellitus, 9 with type II, 7 with impaired glucose tolerance and 10 healthy controls were exposed to a cold pressor test. Plasma concentrations of beta-thromboglobulin (index of platelet activation) and of fibrinopeptide A (index of thrombin formation) were measured before and 15 minutes after forearm immersion in melting ice. Basal levels of both proteins were significantly elevated (p less than 0.02) in the combined group of patients with diabetes and impaired glucose tolerance. While in healthy controls cold exposure failed to modify plasma concentration of either protein, obvious changes occurred in the diabetic and impaired glucose tolerance groups. In the combined patients group, beta-thromboglobulin levels decreased from 1.37 +/- 0.44 nmol/l to 1.03 +/- 0.39 (mean +/- SD, p less than 0.01), after the cold test, possibly in consequence of enhanced vascular permeability; while fibrinopeptide A levels increased from 1.52 +/- 1.03 nmol/l to 3.45 +/- 4.19 (p less than 0.02). The degree and pattern of abnormalities observed in basal as well as stimulated levels of fibrinopeptide A differed somewhat among the three groups of patients. These studies indicate that, in the basal state, patients with diabetes or simple carbohydrate intolerance are more susceptible than controls to platelet activation and that after stress thrombin formation can occur although some variability exists among and within groups of patients. The consequences of such increased thrombotic activity may have a bearing on the pathogenesis of large vessel disease, a complication common to diabetes and impaired glucose tolerance. PMID- 2970692 TI - Experimental and clinical angioplasty with a laser probe fiberoptic catheter system. AB - The ablation of atherosclerotic tissue by laser energy was a promising new idea in the treatment of atherosclerotic vascular diseases. Initial studies showed a lot of beneficial effects but also a lot of complications like perforations and thermal injuries. The introduction of a new fiber optic system with a metalcapped quartzfiber, changing the laser energy into thermal energy, showed significant improvements. First animal studies revealed a higher success rate and a significantly decreased risk of perforation. Based on these results first clinical studies in the femoro-popliteal region were started. Meanwhile 129 patients with stenoses or occlusions of the femoral arteries have been treated with this new fiber optic system under fluoroscopic control. Patency rate was 93% for stenoses and 89% for total occlusions. In all cases the procedure was completed by a balloon angioplasty. Reinvestigations revealed a cumulative one year-patency rate of 77%. This is well comparable to long-term results of balloon angioplasty. First clinical applications of this so called "hot-tip" in coronary arteries proved the possibility of reopening stenoses or occlusions in the small vessels. Still further improvements in flexibility and steerability of the catheter systems have to be made, before this method can become routine too. By further improvement laser angioplasty will find its place besides balloon angioplasty and surgery in the treatment of the atherosclerotic vascular disease. PMID- 2970693 TI - Laser recanalization: a basic and clinical perspective. AB - Inspite of the effectiveness of PTCA and surgery in the treatment of occlusive vascular diseases, still a number of patients remain untreatable by these methods. Laser light with its ability to ablate tissue and to be delivered even in small distal branches of the vascular tree seemed to offer a new perspective. The interaction between laser light and tissue is determined by special physical characteristics. Up till now none of the available laser systems are optimal for application in the cardiovascular system, but still many of them have been effective clinically. One common complication was the high perforation rate by the use of flexible quarz fibers which could not be visualized by fluoroscopy. The addition of a small metal ring and later-on of a metal cap covering the end of the fiber showed significant improvements. Not only could those fibers be visualized by a fluoroscopy, but could convert laser energy into thermal energy. The perforation rate was decreased and a greater size channel created. Lately the introduction of a small saphir window at the tip of the metal cap offered the combined application of light and thermal energy. Animal studies demonstrated these advantages with a low perforation and a high penetration rate of atherosclerotic material. Clinical studies also revealed good results and a significantly lower perforation rate with the metal capped probe. Using the probe with a small saphir window at its tip, the success rate in totally occluded peripheral vessels, not treatable by conventional PTCA, was 82%. As the newly created channel showed a sufficient diameter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970694 TI - Percutaneous laser angioplasty in the treatment of peripheral vascular disease. AB - For a long time the treatment of peripheral vascular diseases mainly was a surgical one. Improvements in drug therapy and success of PTCA have increased the interest in a more conservative therapy. So laser promised to be another tool in the percutaneous treatment of peripheral vascular diseases. First successful experiments were followed by the experience of technical problems and side effects of this new method. First clinical studies in occluded human femoral arteries were started at Stanford University in 1983. The results, published in 1985, showed that occlusive lesions in this vascular region could be successfully reopened by a percutaneous approach. But in cases of severe calcification the danger of perforation or dissection was significantly increased. Moreover the newly created channel proved to be too small for a sufficient revascularization, so that this method had to be combined with balloon angioplasty. By the development of the metalcapped "hot tip", laser energy was totally changed into thermal energy. By this new system greater channels of 2 up to 2.5 mm could be created and by the special design of the metalcap thermal injury to the arterial wall could be decreased. Nevertheless perforation occurred also with this system. Moreover, additional balloon angioplasty was still necessary in proximal segments of the femoral artery. Futural developments of greater metal heads may lead to the exclusive use of the "hot tip" especially in more distal parts. Longterm results will show, if laser angioplasty is comparable to other conventional methods like balloon angioplasty or surgery. There is still a long way to go, until the ideal of reopening atherosclerotic stenoses only by laser energy can be reached. PMID- 2970695 TI - Intraoperative laser coronary angioplasty. AB - After the first intraoperative application of an Argon-laser by Choy in 1983, a special trial on laser angioplasty was started in Houston in 1985. Under the indications of tandem stenoses, diffuse atherosclerotic alteration or peripheral stenoses, laser endarterectomy was performed with a hand-held CO2-laser in addition to conventional bypass surgery. Between January 1985 and December 1986, 20 patients were treated with this combined method. Laser endarterectomy was successful in 37 out of 40 diseased vessels. After an interval of one week to 6 months after operation, 18 patients underwent a reangiography, showing a patency rate of 76% in the lased arterial segments. This result is comparable to conventional endarterectomy or intraoperative balloon angioplasty in similar cases. Further improvements can be expected by modifications in antiplatelet drug therapy. PMID- 2970696 TI - [The economic sequelae of mandibular fractures]. PMID- 2970697 TI - [Characteristics of the reaction to disease and its treatment in patients at an orthodontic clinic]. PMID- 2970698 TI - [The possible use of glass ceramic materials in orthodontics]. PMID- 2970699 TI - [The role of ethics and deontology in the success of orthodontic treatment of disabled, middle-aged and elderly patients]. PMID- 2970700 TI - [The development of oral surgical care in Tadzhikistan]. PMID- 2970701 TI - [The need for hospital care of patients with inflammatory processes in the maxillofacial area (based on data from the Moldavian SSR)]. PMID- 2970702 TI - Plasma atrial natriuretic factor and subarachnoid hemorrhage. AB - Hyponatremia is common following aneurysmal subarachnoid hemorrhage and has been linked to the syndrome of inappropriate secretion of antidiuretic hormone. However, the demonstration of volume depletion and natriuresis in some patients has suggested that salt wasting is a more likely etiology. Atrial natriuretic factor appears to play a role in both central and peripheral regulation of sodium homeostasis. To investigate the behavior of circulating atrial natriuretic factor following subarachnoid hemorrhage, we studied 25 patients with intracranial aneurysms: 21 after acute subarachnoid hemorrhage and four without evidence of recent rupture. Atrial natriuretic factor was measured by radioimmunoassay of extracted plasma (normal value, 20.8 +/- 24.6, mean +/- 3 SD). Mean +/- SEM plasma atrial natriuretic factor concentration was elevated to 84 +/- 25 pg/ml on Day 1, rose to 134 +/- 29 pg/ml on Day 3, and fell to 86 +/- 17 pg/ml by Day 7 after subarachnoid hemorrhage (p less than 0.01). In two patients (9.5%) who developed hyponatremia after aneurysm rupture, plasma concentrations were no different from that in the group as a whole; concentrations in patients with no evidence of recent subarachnoid hemorrhage were not elevated. Neither fluid administration nor timing of surgery could account for the elevated concentrations. We conclude that concentrations of circulating atrial natriuretic factor are elevated after subarachnoid hemorrhage but do not solely account for the accompanying hyponatremia. PMID- 2970703 TI - [Protein metabolism during the growth of hybridoma cells]. AB - Protein metabolism during the growth of mouse hybridoma, producing monoclonal antibodies to phage lambda has been studied. The rate of protein accumulation Va at any time of growth was equal to that of protein synthesis Vs minus the degradative rate Vd and protein secretion rate Vsecr (Vn = Vs--Vd--Vsecr). Cellular protein metabolism was described in terms of the relationship 6.3% hr = 8.1%/hr--1%/hr for the middle of exponential growth phase, and in terms of the relationship 1.4%/hr = 2.9%/hr = 0.8%/hr--0.7%/hr for the early stationary phase. The results obtained enabled us to conclude that the synthesis of cellular proteins may be the main process regulating the protein metabolism during hybridoma cell growth. PMID- 2970705 TI - [Doxazocin]. PMID- 2970704 TI - [Sarcoplasmic reticulum Ca2+-ATPase reconstructed into low-permeable proteoliposomes]. AB - The kinetic characteristics of Ca2+-ATPase reconstructed into proteoliposomes were studied with fluorescent probes. Reconstruction was made using purified resin XAD-2. The data obtained evidence for an electrogenic character of the reconstructed Ca2+-ATPase activity. PMID- 2970706 TI - Intra-arterial ultrasonic imaging for recanalization by spark erosion. AB - Presently several new methods are being developed to recanalize obstructed arteries during catheterization. Intra-arterial high frequency ultrasonic imaging may be used as a guidance for these new techniques. Spark erosion is a new obstruction removal technology. Experiments have shown that this method can be applied in a selective way. An ultrasonic intra-arterial imaging system allows for the proper indication of the spark erosion catheter relative to the obstruction. The first in vitro results of this study illustrate that integration of catheter tip imaging and spark erosion is possible. PMID- 2970707 TI - Stability of flow patterns in the in vivo post-stenotic velocity field. AB - Pulsed Doppler velocimetry was used to construct velocity profiles which demonstrated jet flow and recirculation in the poststenotic velocity field of canine peripheral arteries in vivo. Maximal spectral broadening in near midline velocity waveforms was used to assess turbulent type flow disturbance. A doubling or halving of mean volume flow, in iliac and carotid arteries, respectively, had no effect on the spatial relationship of these flow patterns. Peak jet velocity was changed less, and end diastolic velocity was changed more than expected. Magnitude of turbulent type flow disturbance was reduced by halving but not doubling flow. Effects of changes in pulsatility and heart rate are discussed. It is suggested that similar variation in maximum and end diastolic jet velocities and poststenotic spectral broadening will occur in patients undergoing noninvasive diagnostic tests for arterial disease. This will reduce the resolution of methods based on absolute levels of these parameters. PMID- 2970708 TI - Renal calculi-urate as a urokinase inhibitor. AB - The formation of renal calculi is one of the most widely studied urinary ailments. Spectrophotometric analysis of urinary inhibition on the urokinase/plasmin system revealed a significant difference between subjects with and without renal calculi (P less than 0.001). The percentage urokinase/plasmin inhibition in the two groups was 77.1% for those with, and 47.1% for those without renal calculi. Because of the significant (P less than 0.015) positive correlation (r = 0.762) between the percentage inhibition of urokinase/plasmin and urinary urate concentration, the inhibition of urate on urokinase/plasmin and on plasmin and urokinase was investigated. The urokinase/plasmin system was inhibited up to 94.4% with 0.5 mM urate. Inhibition occurred with both low and high molecular weight urokinases. In order to determine which enzyme of the complex was inhibited, each was investigated independently and a 50% inhibition of urokinase activity was obtained with 4 mM urate. It was found that the urine of stone formers contained a higher concentration of urate than the urine of healthy subjects and that at the same time there was a decrease in urinary urokinase activity in these patients. PMID- 2970709 TI - Complete regression of skeletal metastases from hypernephroma after angioinfarction and medroxyprogesterone acetate therapy. PMID- 2970711 TI - Ventricular versus atrial natriuretic factor. PMID- 2970710 TI - Hormonal modulation of macrophage clearance of IgG-sensitized cells. PMID- 2970712 TI - Cutaneous lymphosarcoma of helper/inducer T-cell origin in a calf. AB - A calf with a lymphoproliferative disease resembling human mycosis fungoides, had multiple, ulcerated skin nodules with enlarged regional lymph nodes. Tumor cells were 10-15 micron in diameter and had round to oval nuclei without indentations. Mitoses were regularly present. Pautrier's microabscesses were in the epidermis, and T-cell areas of regional lymph nodes were infiltrated by neoplastic cells. Demonstration of the T-helper/inducer origin of neoplastic lymphocytes suggested classification in analogy with human mycosis fungoides. PMID- 2970713 TI - Equine post anaesthetic myositis: muscular post ischaemic hyperaemia measured by laser Doppler flowmetry. AB - Measurements of muscular microcirculation in horses anaesthetised with halothane were performed by laser Doppler flowmetry. Variations of microcirculation in the compressed and uncompressed triceps brachii were measured when horses were positioned in dorsal recumbency after a prolonged period in lateral recumbency. A significant post ischaemic hyperaemia was recorded in horses which developed a post anaesthetic myositis. PMID- 2970714 TI - Bovine interleukin 2: production by an E-rosette-defined lymphocyte subpopulation. AB - E-rosette-separated bovine peripheral blood lymphocyte subpopulations were examined for ability to produce interleukin 2 (IL 2). Sequential E-rosetting techniques resulted in three T-cell subpopulations and a non-T population. Separated cells were stimulated with Con A and the resulting culture supernatants were assayed for IL 2 activity on IL 2-dependent cells. The bovine T-cell subpopulation which rosetted with both neuraminidase-treated and 2 aminoethylisothiouronium bromide (AET)-treated erythrocytes was found to produce significantly more IL 2 than the other T-cell subpopulations or the non-T population. These results suggest that this population may have a T-helper cell function. IL 2-dependent cells were found to be predominately T-cells by E rosetting, were lymphoblastoid in appearance and surface immunoglobulin negative. Conditioned media containing IL 2 were used to demonstrate cytotoxic T-cell activity against allogeneic lymphocytes in peripheral blood lymphocytes. PMID- 2970715 TI - Adenovirus type 5 and adenovirus type 12 recombinant viruses containing heterologous E1 genes are viable, transform rat cells, but are not tumorigenic in rats. AB - Two sets of adenovirus type 5 (Ad5)-adenovirus type 12 (Ad12) recombinant viruses were constructed and analyzed. In one case the Ad12 E1A, E1B, or E1A plus E1B genes were substituted for the corresponding Ad5 genes in the Ad5 chromosome. The second set contained the Ad5 E1A, E1B, or E1A plus E1B genes in place of the cognate Ad12 genes in the Ad12 chromosome. The hybrid viruses were all viable and expressed the appropriate E1 antigens. They were able to transform secondary rat fibroblasts, but at reduced efficiency as compared to either parental virus. Fibroblasts transformed with the recombinant Ad5 virus carrying the Ad12 E1A plus E1B genes were tumorigenic in newborn, syngeneic rats. Some of the cell lines transformed with the Ad5 virus containing the Ad12 E1A gene were tumorigenic but none of the recombinants with the Ad12 E1B gene was able to induce tumors in this assay. Although Ad12 was tumorigenic, none of the Ad5 or Ad12 recombinant viruses induced tumors in newborn rats injected either intracerebrally or subcutaneously with virus particles. PMID- 2970716 TI - [50 anniversary of medical staff internship in the Leningrad Military District]. PMID- 2970717 TI - [Comparative characteristics of hemagglutinins of influenza A viruses with antigenic structure Hsw1N1 isolated from man and animals]. AB - Competitive radioimmunoassay was used to study the antigenic composition of hemagglutinin of Hsw1N1 viruses isolated from man in comparison with hemagglutinin Hsw1 of influenza virus of swine and ducks. The data of oligonucleotide analysis of the 4th RNA segment coding for hemagglutinin in these viruses are presented. It has been shown that in Alma-Ata, 1984-1985, influenza viruses Hsw1N1 were isolated with the antigenic structure of hemagglutinin and with the hemagglutinin gene identical with those of the classical influenza virus of swine A/Swine/Iowa/15/30 but differing from virus A/New Jersey/8/76. PMID- 2970718 TI - [Suppressive action of the influenza virus on the cellular immune reaction]. AB - Experiments in mice using syngeneic transfer of spleen cells and intraperitoneal inoculation of cyclophosphamide, levamisole, hydrocortisone, and anti-0 serum demonstrated that influenza virus may inhibit T-suppressors from the first days of infection. Then the T-helper system of lymphocytes is inhibited and the capacity of the animals to show delayed type hypersensitivity is reduced. In this period, higher activity of T-helpers is observed. It is assumed that inhibition of the T-helper system is associated with the affection by influenza virus of the cells which activate T-helpers. PMID- 2970719 TI - [Effect of hemosorption on the anti-chalone properties of the blood serum of patients with psoriasis]. PMID- 2970720 TI - [Status of the autonomic nervous system and immunological reactivity in patients with eczema during reflexotherapy]. PMID- 2970721 TI - [Sensitization of workers in the leather-processing industry to occupational irritants]. PMID- 2970722 TI - [Bladder exstrophy--anterior osteotomy of the pelvis--a new surgical technic for facilitating stabilization of the pelvis and abdominal closure]. AB - The reconstruction of a stable anterior pelvic ring and the tension-free soft tissue closure is the key to a successful reconstructive surgery of the ectopic bladder. Instead of the commonly practised iliac osteotomy to achieve this goal we postulate the bilateral anterior osteotomy of the superior ramus of the pubic bone. The operative technique of this new method is described. The early results of four cases are discussed. PMID- 2970723 TI - [Histopathological and histobacteriological research on failures in root canal treatment]. PMID- 2970724 TI - [Experimentally induced osteoplasia in the root canal of canine teeth after application of the hydrogel poly(2-hydroxyethyl methacrylate) and collagen]. PMID- 2970726 TI - [A method for the scanning electron microscopic assessment of initial plaque formation in vivo]. PMID- 2970725 TI - [Chemotaxis study of peripheral granulocytes in patients with adult periodontitis]. PMID- 2970727 TI - [Correction of open bite on the mandible]. PMID- 2970728 TI - [Effect of the processing technic on amalgam creep]. PMID- 2970729 TI - [Evaluation of the success of conservative and surgical therapeutic methods in temporomandibular fractures]. PMID- 2970730 TI - [Fluoride content of the deciduous enamel in amelogenesis imperfecta hereditaria]. PMID- 2970731 TI - [Rhinophyma: pathological anatomy and actual therapeutic criteria]. PMID- 2970732 TI - [Gingival model. Important carrier of information in preparation of prosthesis]. PMID- 2970733 TI - [Hydrocolloids--volume stability and elastic rebound of different media]. PMID- 2970734 TI - [Crowns and partial crowns of new porcelain system]. PMID- 2970736 TI - [Glass ceramic. Possibilities and limits]. PMID- 2970735 TI - [Computer in dental laboratory--yes or no?]. PMID- 2970737 TI - [Right- or left-handedness. Ergonomic resolution. Rotary instruments and interesting ways of cutting]. PMID- 2970738 TI - [Does Doppler blood flow measurement and the fetal actogram proven advantages over the classical fetal monitoring in pregnancy with the aid of cardiotocography? ]. PMID- 2970739 TI - [Patients with chronic viral hepatitis as sources of infection]. AB - Dynamic observation on 126 foci of infection formed by patients with manifest forms of chronic hepatitis B, 41 foci of chronic hepatitis of unknown etiology, and 37 foci formed by chronic "healthy" carriers was made. In the foci of type 1 the epidemic process developed intensively and was manifested mainly by HBsAg carriership in persons having had contacts with the patients. During the period of observation 43.0% of new cases of infection were detected. In the foci of types 2 and 3 the frequency of contacting infection was not different from that in the control group of the population. PMID- 2970740 TI - [Prevention of mumps in preschool institutions using a live mumps vaccine made from strain L-3]. AB - The effectiveness of the emergency vaccinal prophylaxis of epidemic parotitis was studied in 19 children's day-care centers. As revealed in this study, the immunological effectiveness of vaccination did not depend on the age of vaccinees, but sharply decreased if live parotitis vaccine contained less than 10,000 HADU50 per immunization dose. After a single administration of the vaccine 91.1 +/- 0.98% of children were found to produce mumps antibodies. The immunization of children with live parotitis vaccine prepared from strain l-3 immediately after the first case of parotitis had been registered proved to be a highly effective measure. The coefficient of epidemiological effectiveness was 96.4%. PMID- 2970741 TI - [Epidemiological characteristics of postoperative wound infections in a trauma center hospital]. AB - The occurrence of cross infections in osteomyelitis patients has been proved by clinical, microbiological, and epidemiological methods. The patients are infected mainly in the dressing room for patients with purulent processes; the working conditions in this dressing room do not preclude the possibility of cross infections. PMID- 2970742 TI - [A natural focus of icterohemorrhagic leptospirosis in the floodplain of the lower Don and means for its eradication]. AB - An active natural focus of icterohemorrhagic leptospirosis has been detected in the area of fish-breeding ponds in Rostov Province, where the intensive epizootic among the population of Norway rats is observed the whole year round (574 animals have been examined, 56 cultures have been isolated). The epizootic process reaches its highest intensity in autumn (the proportion of infected animals exceeds 50%). This natural focus in the area of fish-breeding ponds is epidemiologically dangerous. The limitation of its infectious potential is possible by means of poisoned baits. PMID- 2970743 TI - [Results of a study of a live intranasal influenza vaccine in the immunization of children 3 to 15 years old]. AB - A total of 10,971 children aged 3-15 years were immunized with live influenza vaccine (the variant intended for children), type A (H1N1). The vaccine proved to be nonreactogenic and produced no harmful effect on the child organism. The genetic stability of the vaccine strain and its high immunogenic activity were shown. PMID- 2970744 TI - [Prophylactic efficacy of reaferon in viral hepatitis A and acute respiratory infections in children]. AB - Reaferon, the analog of human alpha 2-interferon obtained by gene engineering techniques, was studied with a view to its use for the prevention of hepatitis A. The study involved children of preschool age in Tashkent. In a strictly controlled trial children aged 2-6 years received the preparation orally in a dose of 1 X 10(6) I. U. or the diluent alone used as placebo. The preparation was administered to 1,100 children and the placebo to 1,078 children. The preparation and placebo were administered twice a week for two months. On the whole, during that period hepatitis A morbidity in both test and control groups of children was the same (5.1% and 4.9% respectively), but among children of nursery age receiving Reaferon the incidence of hepatitis A and acute respiratory viral infections was lower than among those receiving the placebo, though this difference was statistically significant only for cases of acute respiratory infections. PMID- 2970745 TI - [The dynamics of alcoholic psychoses in recent years in the intensified campaign against drunkenness and alcoholism]. AB - An analysis of hospital admissions for alcoholic psychoses made among males in an industrial city over the period of July-December 1984 and the corresponding period of 1985 showed that the number of admissions in 1985 decreased by one third largely at the expense of people relatively intact from the social and clinical standpoint. PMID- 2970746 TI - [The prognostic selection of subjects with neurological manifestations of lumbar osteochondrosis during mass prophylactic examinations of the population (screening)]. AB - Several types of tabulated and computerized screening systems have been developed for the selection of subjects at high risk of the development or a relapse within 1-3 years of marked lumbosacral pain involving a temporary disability. The systems are intended to be used during wide-scale prophylactic medical examinations but may also be employed on an individual basis. Some of the proposed systems are based on a self-interview (7-9 questions), others take into consideration objective neurological findings. The authors present a simple calculation table to assess the probability of a relapse of vertebrogenic lumbosacral pain within 12 months. The prognostic coefficients were calculated with the help of modified Bayes' method. PMID- 2970747 TI - [The course of neurological manifestations of lumbar osteochondrosis and the quantitative assessment of prognostically significant factors]. AB - On the basis of computer-assisted processing of an archive of formalized histories of 2187 patients with neurological manifestations of lumbar osteochondrosis the author has revealed a number of regularities characterizing the time-course of this disease. A comparative quantitative assessment of the prognostic value of a number of history, clinical and paraclinical parameters is presented. Tabulated and computerized systems predicting the course of the disease in a given patients are described. Data obtained during 1-3 year long follow up are presented. PMID- 2970748 TI - [The pathomorphosis of alcoholic delirium]. AB - The authors have examined case histories of 402 patients with delirium tremens treated in 1897-1931, in 1955-1970 and in 1971-1985. An analysis has demonstrated a reduction in the frequency of visual hallucinations of zoological contents and an increase of erotic hallucinations. The duration of the psychotic state in different time periods has been determined. Changes in manifestations of emotional disturbances are described. PMID- 2970749 TI - [The medico-psychological situation in Leningrad 1941-1945 (somatopsychic relationships)]. PMID- 2970750 TI - [Dynamics of the immune status and the role of the T-lymphocyte subpopulation in the acute period of craniocerebral injuries]. AB - The author studied the blood immune values (T-, B-lymphocytes, T-helpers, T suppressors, immunoglobulins M, G, A) in 84 patients with craniocerebral trauma (CCT) of different severity in the acute period. Changes in the values according to the severity of the trauma and the time of examination were revealed. In severe CCT the organism's defence forces were found to be reduced on the 2nd-3rd week according to the values of T-lymphocyte subpopulations, which may be the cause of secondary inflammatory complications. PMID- 2970751 TI - Vanadate stimulated NADH oxidation in sarcoplasmic reticulum membrane. AB - Vanadate stimulated NADH oxidation was detected in sarcoplasmic reticulum membrane preparations. The reaction showed enzymatic character, with half maximal activating concentration of 1.2 mM vanadate and maximal NADH oxidation 50 nmol/mg protein/minute. Acidic pH, micromolar free Ca2+ concentration and decavanadate addition increased the rate of NADH oxidation. The described enzyme activity is similar to the ones observed in erythrocyte liver and cardiac plasma membranes. The vanadate stimulated NADH oxidation in sarcoplasmic reticulum preparations does not seem to originate from other contaminating membrane elements. The presence of this enzyme activity in the sarcoplasmic reticulum should be taken into consideration when planning experiments with vanadate, especially when measuring ATPase activity through NADH oxidation with coupled enzymatic assay. PMID- 2970753 TI - [Safety provisions for recreational flying or sport with a hang-glider]. AB - The Act. n. 106 of 25th March 1985 had defined the specifications of the particular aircraft designed for hobby or sport flying as is the hang-glider. It has also provided for the issue, within six months, of special regulations aimed at "checking the psycho-physical fitness required in handling" such aircraft in additions to the technical knowledge and the information about traffic, safety, insurance regulations relevant to the matter. However, the patent default of the legislator causes the protection of hobby and sport practice of hang-gliding to be either wholly inadequate or ruled by ambiguous regulations. If, instead, the present law in force is referred to, it is deemed that--while waiting for the regulations provided for by n. 106 Act.--the Aereo Club of Italy may define as "agonistic" the practice of "hobby or sport flight" so that the checking of the "specific" fitness required by such sport comes into operation in compliance with the State Decree of 18th February 1982. PMID- 2970752 TI - [Results of infiltration of lower lumbar posterior articulations in lumbago]. PMID- 2970754 TI - [Clinical use of pantethine by parenteral route in the treatment of hyperlipidemia]. AB - Recent investigations have confirmed the effectiveness and the excellent tolerability of pantethine, a derivative of pantetheine, an essential part of the acetylation coenzyme CoA, administered P.O., in normalizing the blood lipid concentrations of patients with hyperlipidemias. A group of 18 patients with hyperlipidemias (9 M, 9 F), with an average age of 52.6 years, was submitted to pantethine parenteral treatment. After a 20 days wash-out, pantethine (400 mg/day; BID) was administered intramuscularly, for 20 days. Total cholesterol, triglycerides, HDL-cholesterol, apo A-1 and B lipoprotein, uric acid in serum, glycemia, CBC, B.U.N., creatininemia, E.S.R., SGOT, SGPT, bilirubinemia, cardiac frequency, blood pressure and body weight were controlled before and after treatment. The drug showed to have a therapeutic effectiveness by a rapid and significant improvement in the blood lipid pattern with reduction of total cholesterol, triglycerides and apo-B lipoprotein and increase of HDL-cholesterol and apo A-1 lipoprotein. The tolerability of pantethine at the stated dosage and mode of administration was invariably excellent, with non complaints or visible side effects imputable to the test drug. BUN, creatininemia, glycemia, SGOT, SGPT, bilirubinemia, E.S.R., CBC, cardiac frequency and blood pressure readings showed no noteworthy changes throughout the study. PMID- 2970755 TI - [Renal colic: behavior of urinary parameters in the assessment of lithogenic risk]. AB - In patients with renal colic we studied lithogenic urinary risk factors before and after the stone passage. We showed abnormalities in water, electrolytes and other substances excretion due to retention and metabolic disorders. The effects more pronounced is on urinary sodium, calcium, magnesium and ammonium. Citrate behaviour suggests a transient intracellular acidosis. PMID- 2970756 TI - [Acquired immunodeficiency syndrome: epidemiological, clinical and immunological findings in risk groups in Parma]. AB - Between August 1985 and June 1987, 809 subjects at risk for AIDS have been studied. 231 (28.5%) were seropositive for human immunodeficiency virus (HIV) antibodies. The seropositivity rate was 41% among drug addicts, 20.5% among homosexual/bisexual males, 19.7% among sexual partners of seropositive individuals. None of 62 subjects belonging to the health care personnel who interacted with seropositive patients and none of the 26 relatives of HIV infected subject, have been found to be seropositive. Moreover the HIV seropositivity in the population of Parma was only 0.01%. Among the seropositive subjects, 155 (67.1%) were asymptomatic; 2 (0.8%) showed acute infection (a mononucleosis-like syndrome in both, associated with aseptic meningitis in one); 57 (24.6%) had PLG, 7 (3.4%) ARC, 9 (3.8%) full-blown AIDS (8 of these latter are dead). PMID- 2970757 TI - [A case of rupture of the testis]. AB - A case of traumatic rupture of the testis is presented in which the conservative surgery was successful. The results of the ultrasonographic evaluation and of the scanning with Tc 99, executed long after the operation, are presented. The importance of an early explorative operation in all cases of post traumatic scrotal swelling is stressed. On account of the youth of most patients, the importance of a conservative surgical approach is emphasized. PMID- 2970758 TI - [Familial malignant melanoma and its relation to dysplastic nevus syndrome: considerations on 6 clinical cases]. AB - Two cases of familial expression of malignant melanoma are presented, with regard to three patients recently operated on for this neoplasm. The authors review the literature existing on this subject and make important comments about some hereditary factors probably predisposing to malignant melanoma; among these characteristics are the pigmentary traits of skin, adnexa and iris, the so-called "dysplastic" or "atypical" moles, the cutaneous sensitivity to the sun. PMID- 2970759 TI - [Hepatic abscesses secondary to peritonitis: puncture and ultrasonically guided transparieto-hepatic drainage]. AB - The Authors describe one case of duplex and separated hepatic abscesses secondary to perforated appendicitis and peritonitis. A contemporary literature review confirms the actual rarity of this entity. The patient was treated by percutaneous and transhepatic puncture and drainage of abscesses under sonographic control in two different stages. Material and methods of this technique are herein reported, in particular those concerning the management of the smaller of the two abscesses, located posteriorly into the lower segment of the right lobe of the liver, having a more difficult approach. The therapeutic results were excellent: either few days later percutaneous treatment than two months later after the patient was discharged, the clinical and sonographic controls confirmed a successful recovery. PMID- 2970760 TI - [Various aspects and problems in surgery of the parotid gland: experience in 70 cases]. AB - The pathology and the relative surgical treatment, performed in all the swellings of the parotid gland observed in our clinic, are reported. Particularly the most important topics and problematics, both of short and long term, connected to the surgical therapy carried out, are described. Finally the actual diagnostic aspects and the necessity of an accurate preoperatory staging are discussed. PMID- 2970761 TI - The pharmacokinetics of fentanyl, sufentanil and alfentanil: a comparative review. PMID- 2970763 TI - Prenatal diagnosis of abdominal wall defects. AB - The antenatal diagnosis of omphalocele and gastroschisis is briefly reviewed, together with the major differences between the two lesions. The frequency of associated anomalies in cases of omphalocele is stressed. The controversy regarding the mode and timing of delivery is outlined. PMID- 2970762 TI - Tracing of lateral pterygoid muscle-related neurons in the trigeminal brainstem nuclei in the rat. AB - Retrograde transport of fluorescent tracers (diamidino yellow and true blue) was used to study the arrangement of brainstem neurons innervating the lateral pterygoid muscle in the rat. The lateral pterygoid motoneurons were located in the dorsolateral (jaw-closing) part of the trigeminal motor nucleus with clear somatotopy in the caudal part of the nucleus. No muscle-related neurons were present in the mesencephalic trigeminal nucleus. Histological examination of serial sections of lateral pterygoid muscles confirms the notion that, at least in the rat, this muscle is devoid of muscle spindles. PMID- 2970764 TI - Abdominal wall defects: neonatal surgical problems. AB - The advantages accruing from the prenatal diagnosis of gastroschisis and omphalocele are outlined. The surgical management of each defect is discussed, including details of the various surgical options available at the time of the repair. Seventy-five cases with abdominal wall defects born between 1970 and 1984 without an antenatal diagnosis having been made are compared with 8 recent cases in whom an antenatal diagnosis was made. The early results suggest that antenatal diagnosis improves survival. PMID- 2970765 TI - Early diagnosis of intrauterine growth retardation by Doppler ultrasound. AB - The significance of intrauterine growth retardation (IUGR) and the methods currently available for assessment of fetal growth are briefly reviewed. The use of Doppler ultrasound assessment of the flow velocity waveform (FVW) of the uteroplacental waveform at 16-18 weeks of gestation is outlined. A pilot study of 127 pregnancies suggested that the sensitivity of the test was 67% and the specificity 65%. The prediction value of a negative test was 92%. Six of the 14 cases of IUGR had a normal FVW, but 3 of these had asymmetrical morphology. Four of the 8 IUGR cases with an abnormal FVW required delivery by cesarean section. The FVW also appears to be a valuable predictor of pregnancy-induced hypertension and fetal asphyxia. PMID- 2970766 TI - Unstable angina pectoris. Experience with an acute revascularization program. AB - One year's experience of an acute revascularization program for unstable angina pectoris is presented. In the total material of 63 patients, significant coronary artery stenosis was found in 61 and normal coronary arteries in two. Coronary artery bypass grafting was performed in 39 patients (62%) and percutaneous transluminal angioplasty in nine (14%). There were two perioperative myocardial infarctions and one hospital death. Thirteen patients were not eligible for revascularization. At follow-up (14-26 months) 60 patients were still alive (95%). In the revascularization group, one patient had died, but of the remaining 46 patients, 30 (65%) were free of angina pectoris, and in addition six had only minor symptoms. PMID- 2970767 TI - Plasma atrial natriuretic peptide in cardiac transplant recipients. A prospective study. AB - Cardiac transplantation in 10 patients with congestive heart failure resulted in reduction of high plasma concentrations of atrial natriuretic peptide (ANP), preoperatively five-fold above normal, to a level two-fold above normal, which was maintained throughout a 12-week follow-up period. Cardiac function was normalized in all patients. Transient increases in plasma ANP, in four cardiac recipients 3-10-fold their basal levels, could neither be related to rejection episodes nor to cardiac dysfunction, but rather to signs of fluid and sodium retention. High plasma ANP levels in cardiac transplant recipients suggest that the capacity to secrete ANP is preserved in the transplanted heart. PMID- 2970768 TI - Atrial natriuretic peptide, angiotensin II and aldosterone in plasma in chronic glomerulonephritis during basal conditions and during exercise. AB - Atrial natriuretic peptide (ANP), angiotensin II (AII) and aldosterone (Aldo) in plasma were determined at supine rest in 16 normotensive and 9 hypertensive patients with chronic glomerulonephritis and in 18 control subjects (Study 1). In addition, 12 of the normotensive, 7 of the hypertensive patients and 11 of the control subjects were studied with the same parameters after an exercise test (Study 2). Study 1 showed that supine ANP, AII and Aldo did not differ significantly between the groups. In Study 2, ANP increased after exercise in the normotensive patients (8.4 vs. 11.4 pmol/l (median), p less than 0.05) and control subjects (7.6 vs. 9.3 pmol/l, p less than 0.02) but not in the hypertensives (7.6 vs. 7.9 pmol/l, p greater than 0.10), and after exercise ANP was increased in the normotensive patients compared with the controls (p less than 0.02). After exercise, an enhanced increase of Aldo was found in the hypertensives but not in the normotensive patients compared with the controls, whereas the increase of AII did not differ significantly between the groups. It is concluded that patients with chronic glomerulonephritis and relatively well preserved renal function do not have major abnormalities of ANP at rest or during exercise. In the normotensive patients, however, ANP increased to a higher level than in the controls, but the difference was small and further studies are needed to define the role of ANP in blood pressure regulation of early stage chronic glomerulonephritis. PMID- 2970769 TI - Experimental study on myocardial salvage by coronary thrombolysis and mechanical recanalization. AB - Salvage of the ischemic myocardium by coronary thrombolysis and mechanical recanalization (simulated angioplasty) was studied in a canine experimental model of acute myocardial infarction induced by coronary occlusive thrombus at the left anterior descending coronary artery. Forty-four open-chest dogs divided into three groups were studied. Group I (n = 15, control group) was observed for 6 hours following the onset of infarct. In group II (n = 14, thrombolysis group), thrombolysis was obtained by intravenous administration of urokinase 2 hours after the onset of infarct. In group III (n = 15, mechanical recanalization group), simulated angioplasty was performed 2 hours after infarct. Coronary reperfusion was continued for 4 hours in groups II and III. The areas of left ventricular risk and infarct were measured by double staining methods with Evans blue dye and triphenyl tetrazolium hydrochloride. There were no significant differences in control blood flow and risk area in the three groups. Myocardial infarct area/risk area was 65 +/- 3% in group I, 45 +/- 1% in group II, and 35 +/ 2% in group III (group I vs II, p less than 0.001; group II vs III, p less than 0.001). Restored coronary blood flow in the left anterior descending artery was 8 +/- 1 ml/min in group II and 14 +/- 1 ml/min in group III (p less than 0.001). The data suggest that coronary mechanical recanalization is more effective than thrombolysis in salvaging the ischemic myocardium in the early phase of myocardial infarction, most probably because coronary blood flow is better restored by mechanical recanalization. PMID- 2970770 TI - Acute coronary occlusion occurring after successful percutaneous transluminal coronary angioplasty: temporal relationship to discontinuation of anticoagulation. AB - The incidence and prognosis of acute coronary reocclusion occurring after patients had left the catheterization laboratory following a successful percutaneous transluminal coronary angioplasty (PTCA) procedure and the temporal relation of this event to the discontinuation of systemic heparin administration were analyzed in a series of 1238 consecutive patients. Acute reocclusion, 1 to 96 hours after successful PTCA, occurred in 22 of 1238 patients (1.8%). Patients undergoing PTCA in the setting of acute myocardial infarction were excluded. Out of 22 patients, 15 had a nonocclusive dissection and four had evidence of small intracoronary thrombus immediately post-PTCA, with no evidence of flow disturbance. Acute reocclusion occurred within 5 hours of heparin discontinuation in 12 patients or while they were receiving inadequate anticoagulation (four patients). In 16 of 22 (73%) patients, acute reocclusion was temporally related to a time of diminished anticoagulation. Redilation was attempted in 14 patients and was ultimately successful in five patients (36%). Ten patients required coronary artery bypass surgery and three patients died. Our findings suggest that acute reocclusion after an initially successful PTCA has a poor outcome and seems to be temporally related to the loss of effective anticoagulation in most of these patients. It is advisable to discontinue heparin infusion at a time when facilities for urgent revascularization are available. PMID- 2970771 TI - Neurohumoral and hemodynamic changes in congestive heart failure: lack of correlation and evidence of compensatory mechanisms. AB - The objective of this study was to assess the hemodynamic and neurohumoral (plasma renin activity, aldosterone, epinephrine, norepinephrine, vasopressin, and atrial natriuretic peptide) determinants of systemic vascular resistance in 35 patients with stable congestive heart failure. In the supine position, although activation of the various neurohumoral systems tended to occur in the same patients, there was little correlation between activation of any of the neurohumoral systems, as reflected by circulating levels, and systemic vascular resistance. There was also little correlation between changes in circulating neurohormones and changes in either mean arterial pressure or systemic vascular resistance in the standing position. Acutely reducing the activity of the renin angiotensin system with the use of captopril did not improve the correlation between other neurohumoral and hemodynamic variables. In fact there was no correlation between the effects of acute captopril therapy and baseline renin values. These results support the concept that activation of one or another vasoconstrictor neurohumoral system varies from patient to patient and that the effects of their activation are tempered by activation of parallel vasodilator systems and by attenuation of neurohormone release and effector organ response. PMID- 2970772 TI - Left ventricular hypertrophy among dark- and light-skinned Puerto Rican men: the Puerto Rico Heart Health Program. AB - Racial groups were assessed by determinations of skin color in 3366 urban men in the Puerto Rico Heart Health Program, an epidemiologic study of coronary heart disease in Puerto Rico. These men participated in a comprehensive cardiovascular examination that included assessment of blood pressure, relative weight, physical activity, cigarette smoking, left ventricular hypertrophy (LVH) as determined by ECG, and measurements of skinfold thickness. They were followed for mortality over the next 6 years. Dark-skinned Puerto Rican men had a higher prevalence of both definite and possible LVH-ECG after stratifying by levels of hypertension, relative weight, physical activity, number of cigarettes smoked, and subscapular skinfold thickness. After multivariate adjustment of these factors, the dark skinned men had approximately twice the prevalence of both definite and possible LVH-ECG as the lighter skinned men. Definite LVH-ECG carries a high mortality risk over the next 6 years as compared to men without definite LVH (fivefold increased risk in lighter skinned and sevenfold increased risk in darker skinned Puerto Rican men). PMID- 2970773 TI - Electrocardiographic diagnosis of exercise-induced left ventricular hypertrophy. AB - To assess the prevalence of physiologic left ventricular hypertrophy and the usefulness of ECG criteria for its diagnosis, we compared ECGs and M-mode echocardiograms from 44 ultraendurance athletes and 20 similarly aged sedentary control subjects. Left ventricular mass was elevated in 25 of 44 (57%) athletes including 17 of 29 (59%) men greater than 134 gm/m2 and 8 of 15 (53%) women greater than 110 gm/m2. The sensitivity and specificity of the three ECG criteria used to diagnose left ventricular hypertrophy were: Sokolow-Lyon voltage (S-V1 + R-V5 greater than or equal to 3.5 mV), 65% and 61%; Romhilt-Estes score (greater than or equal to 4), 16% and 84%; and Cornell voltage (R-aVL + S-V3 greater than 2.8 mV in men and greater than 2.0 mV in women), 8% and 95%, respectively. Left ventricular mass, mass index, posterior wall thickness, chamber diameter, and relative wall thickness were not related to any measurement of QRS voltage. Nonvoltage ECG criteria for left ventricular hypertrophy were rare in athletes. Thus hypertrophy is a common but not universal adaptation to exercise. It is only moderately well detected by standard voltage criteria for left ventricular hypertrophy and is not reflected in nonvoltage criteria. PMID- 2970774 TI - Comparison of left ventricular function and contractile reserve after successful recanalization by thrombolysis versus rescue percutaneous transluminal coronary angioplasty for acute myocardial infarction. AB - To determine how coronary reperfusion affects rest and exercise ventricular function after acute myocardial infarction (AMI), 63 patients with a patent infarct artery after intravenous thrombolytic therapy (lysis) were compared with 27 patients who failed thrombolysis but had successful acute recanalization by percutaneous transluminal coronary angioplasty (PTCA) as a "rescue" procedure. Contrast ventriculography was performed acutely and on day 7. Resting radionuclide ventriculography was performed at 24 hours and repeated with exercise on day 30. There were no differences in global ejection fraction (EF) between the 2 groups during acute contrast ventriculography. However, by 24 hours, the EF had deteriorated in the rescue group (40 +/- 17 vs 49 +/- 11% in the lysis group, p less than or equal to 0.05). No improvement occurred in either group on day 7. By day 30, an improvement in resting radionuclide EF 5.9 +/- 1.9% occurred in rescue patients and the difference between rescue and lysis groups was no longer significant (46 +/- 14 vs 50 +/- 11%, p = 0.12). A normal (greater than or equal to 5%) increase in EF with exercise occurred in 64%, with either normal or exercise-enhanced regional wall motion present in 67% of patients. A significant increase in EF occurred within the rescue group, from 46 +/- 14% at rest to 50 +/- 15% at peak exercise (p less than or equal to 0.0005). The EF increased with exercise from 50 +/- 11 to 58 +/- 15% in the lysis group (p less than or equal to 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970775 TI - Left ventricular function after successful percutaneous transluminal coronary angioplasty for postinfarction angina pectoris. AB - The purpose of this study was to determine if coronary revascularization by balloon angioplasty (PTCA) in patients with postinfarction angina can elicit an improvement of global systolic left ventricular (LV) function. LV function was evaluated in 18 patients with postinfarction angina based on peak aortic blood acceleration measured noninvasively with a continuous wave Doppler velocimeter. Initial Doppler measurements were made 4 +/- 1 days after infarction and just before PTCA and were repeated 24 to 48 hours after PTCA. Patients were divided into 2 groups. Group I (n = 10) had successful PTCA (reperfusion). Group II (n = 8) either had an unsuccessful PTCA or did not undergo PTCA or thrombolytic therapy (no reperfusion). In group I patients, peak acceleration increased from 16 +/- 1 m/s/s just before PTCA to 24 +/- 2 m/s/s 24 to 48 hours after successful PTCA (p less than 0.001). In group II patients, peak acceleration was 17 +/- 2 m/s/s just before PTCA and remained relatively unchanged (18 +/- 2 m/s/s) 24 to 48 hours later. These data indicate that revascularization of coronary arteries in patients with postinfarction angina can elicit considerable improvement of global LV systolic function. PMID- 2970776 TI - Coronary revascularization after intravenous tissue plasminogen activator for unstable angina pectoris: results of a randomized, double-blind, placebo controlled trial. AB - To determine the role of intravenous tissue plasminogen activator (t-PA) in unstable angina, it was compared with placebo in a randomized, double-blind trial. Forty patients with angina at rest and provocable ischemia (pacing induced) had baseline coronary angiography, study drug infusion and then repeat angiography at 20 +/- 9 hours. All patients received diltiazem, nitrates, beta blockers, aspirin and intravenous heparin. During study drug infusion (150 mg over 8 hours), refractory ischemia necessitating emergency bypass surgery (CABG) or coronary angioplasty (PTCA) occurred in 4 of 20 t-PA patients compared with 1 of 20 placebo patients (p = 0.21). Before discharge, revascularization for persistent, provocable ischemia and a residual stenosis greater than or equal to 60% was as follows: t-PA patients, 8 PTCA and 7 CABG; placebo patients, 11 PTCA and 8 CABG (p = 0.39). Quantitative angiographic percent diameter stenosis of the culprit artery at baseline and follow-up was: t-PA 71 +/- 17 and 63 +/- 22; placebo 70 +/- 19 and 67 +/- 22 (difference not significant). However, 3 t-PA patients compared with no placebo patients demonstrated an insignificant (less than 60% diameter) residual stenosis and averted PTCA (p = 0.14). There were no complications of PTCA in the 8 t-PA patients; in contrast, 3 of 11 placebo patients had abrupt closure, necessitating emergency CABG in 2 (p = 0.23). Thus, intravenous t-PA in unstable angina can eliminate the need for PTCA in a few patients, does not appear to decrease the overall or emergency rate of revascularization procedures and may facilitate the safety of PTCA. PMID- 2970777 TI - Administration of enoximone in cardiogenic shock. AB - Thirteen patients in severe cardiogenic shock, persisting despite the use of adrenergic agents, were treated with enoximone, a recently available phosphodiesterase inhibitor. Cardiogenic shock was characterized by low cardiac output (less than 2.5 liter.min-1.m-2), elevated pulmonary artery balloon occluded pressure (greater than or equal to 15 mm Hg), decreased urine output (less than 20 ml.hour-1) and increased blood lactate (greater than or equal to 2.0 mEq.liter-1). Ten patients were mechanically ventilated. A short-term intravenous infusion of 0.5 mg.kg-1 in 20 minutes of enoximone resulted in significant increases in cardiac index (from 1.8 +/- 0.3 to 2.9 +/- 0.3 liter.min 1.m-2, p less than 0.001) and stroke index (from 17.8 +/- 3.3 to 21.9 +/- 5.1 ml.m-2, p less than 0.001) and significant decrease in pulmonary artery balloon occluded pressure (from 21.7 +/- 5.8 to 19.8 +/- 6.0 mm Hg, p less than 0.01) without a consistent change in mean arterial pressure (from 79 +/- 8 to 76 +/- 9 mm Hg, difference not significant). Enoximone administration decreased arterial oxygen tension (from 108 +/- 42 to 94 +/- 36 mm Hg, p less than 0.01) and increased venous admixture (from 12.8 +/- 6.5 to 16.0 +/- 8.0%, p less than 0.01). In 8 patients, a second infusion of 0.5 mg.kg-1 immediately thereafter amplified these changes. All patients but one survived the episode of cardiogenic shock and 5 patients left the hospital alive. These results indicate that the addition of enoximone to adrenergic agents in the treatment of cardiogenic shock can markedly increase cardiac output and stroke volume without substantial effects on arterial pressure. PMID- 2970778 TI - Evaluation of age, gender, heart rate and blood pressure changes and exercise conditioning on Doppler measured aortic blood flow acceleration and velocity during upright treadmill testing. AB - To evaluate the effect of upright exercise on aortic peak flow acceleration and velocity, 60 normal subjects between 15 and 74 years of age were evaluated by continuous wave Doppler during treadmill stress testing using the Bruce protocol. Subjects were divided into 3 age groups, each with 20 subjects: group 1, 21 +/- 4 years of age (mean +/- standard deviation), group 2, 36 +/- 5 years and group 3, 58 +/- 7 years. Periodic measurements of heart rate, blood pressure and Doppler blood flow velocity and acceleration were made before, during and after exercise. Continuous wave Doppler measurements were recorded from the suprasternal notch. The relation between Doppler aortic measurements and age, gender, normal heart rate and blood pressure responses during exercise, and exercise preconditioning, was evaluated. Age alone was significantly related (inversely) to immediate postexercise Doppler aortic peak blood flow peak acceleration (group 1, 55 +/- 15, group 2, 46 +/- 11 and group 3, 36 +/- 9 m/s2, p less than 0.05) and peak velocity (1.1 +/- 0.2, 1.0 +/- 0.2 and 0.8 +/- 0.2 m/s, respectively, p less than 0.01). Gender, heart rate and blood pressure changes during exercise, as well as preconditioning, had no significant effect on these flow characteristics. Consequently, the effects of normal aging must be considered when using Doppler measurements of peak aortic acceleration and velocity in the evaluation of left ventricular function, e.g., to detect or exclude the presence of coronary artery disease. PMID- 2970779 TI - Pattern of left ventricular diastolic filling associated with right ventricular enlargement. AB - Right ventricular (RV) dilatation associated with pressure overload may alter left ventricular (LV) geometry resulting in abnormal diastolic function as demonstrated by a smaller LV diastolic volume for a given LV diastolic pressure. To determine whether abnormalities in LV geometry due to RV dilatation result in abnormalities in the LV diastolic filling pattern, we obtained pulsed Doppler transmitral recordings from 23 patients with RV dilatation with RV systolic pressure estimated to be less than 40 mm Hg (group 1), 18 patients with RV dilatation and RV systolic pressures greater than or equal to 40 mm Hg (group 2) and 33 normal patients. RV systolic pressures were estimated from continuous wave Doppler peak tricuspid regurgitation velocities using the modified Bernoulli equation. Diastolic filling parameters in group 1 patients were similar to normals. In group 2 patient, increased peak atrial filling velocity (76 +/- 14 vs 57 +/- 12 cm/s, p less than 0.001), decreased peak rapid filling velocity/peak atrial filling velocity (1.1 +/- 0.4 vs 1.5 +/- 0.4, p less than 0.01), increased atrial filling fraction (41 +/- 14 vs 30 +/- 10%, p less than 0.01) and prolongation of the atrial filling period (171 +/- 47 vs 152 +/- 39 ms, p less than 0.05) were noted compared with the normal group. RV end-diastolic size and LV end-systolic shape were significantly correlated with the atrial filling fraction in group 2 patients. In patients with RV dilatation and RV systolic pressures greater than or equal to 40 mm Hg, there is increased reliance on atrial systolic contribution to the LV filling volume. PMID- 2970780 TI - Percutaneous transluminal coronary angioplasty in patients with intracoronary thrombus. PMID- 2970781 TI - Long-term follow-up and influence of symptom-free interval on restenosis after repeat percutaneous transluminal coronary angioplasty. PMID- 2970782 TI - New electrocardiographic criteria for right ventricular hypertrophy: a correction for the 1986 study. PMID- 2970783 TI - Influence of enoximone and UDCG-115 on coronary hemodynamics in idiopathic dilated cardiomyopathy. AB - Coronary hemodynamics were studied in 24 patients with idiopathic dilated cardiomyopathy and in 17 patients without any significant heart disease under resting conditions using the argon method. Neither myocardial blood flow normalized for 100 g muscle tissue nor myocardial oxygen consumption per minute (MVO2) or oxygen supply-demand ratio were different between these 2 groups of patients. When enoximone (1 to 2 mg/kg body weight) was given intravenously in patients with idiopathic dilated cardiomyopathy, myocardial oxygen consumption decreased by only 8% (difference not significant), whereas a significant (p less than 0.05) 26% decrease of myocardial oxygen consumption was observed after UDCG 115 (1.25 mg/hour intravenously). However, with both substances the oxygen supply demand ratio significantly increased from 1.46 +/- 0.10 to 1.57 +/- 0.20 (p less than 0.025; enoximone) and from 1.40 +/- 0.08 to 1.56 +/- 0.19 (p less than 0.05; UDCG-115), respectively. It is concluded from these data that (1) resting coronary hemodynamics related to a unit of myocardium are not different between normal and idiopathic dilated cardiomyopathy, and (2) phosphodiesterase inhibitors exert beneficial effects on coronary hemodynamics by improving the oxygen supply-demand ratio. PMID- 2970784 TI - Regional blood flow in congestive heart failure: concept of compensatory mechanisms with short and long time constants. AB - With physiologic stress to the cardiovascular system, some circulatory compensatory mechanisms are designed to restore homeostasis quickly (e.g., sympathetic nervous system activation and the Frank-Starling mechanism). These compensatory mechanisms are not nearly as effective when there is a chronic pathologic stress such as congestive heart failure (CHF). In this circumstance, other mechanisms that operate with longer time constants come into play (e.g., activation of the renin-angiotensin-aldosterone system, myocardial hypertrophy and deconditioning). The most successful chronic drug therapies of CHF are those that are designed to reverse the latter group of compensatory mechanisms, a process that is slow. It takes especially long to reverse those CHF-induced changes in blood vessels and skeletal muscle metabolism that are activated to cope with inadequate delivery of oxygenated blood to working muscles. The concept that compensatory mechanisms have either short or long time constants for activation, effectiveness and reversal may help explain why the improvement in exercise tolerance with effective heart failure therapy lags behind hemodynamic improvement. PMID- 2970785 TI - Reduced subendocardial myocardial perfusion as one mechanism for congestive heart failure. AB - One mechanism for the eventual decompensation of the hypertrophied to the failing heart may involve inadequate myocardial perfusion. In support of this concept are studies in experimental models of both right and left ventricular hypertrophy and failure. These studies demonstrate blunted reactive hyperemic responses to brief periods of coronary artery occlusion, and reduced vasodilation in response to adenosine, when the most severe impairment in coronary reserve occurs in the subendocardium. During exercise, in the hypertrophied and failing heart, the normal coronary vasodilator response is also blunted. Although endocardial/epicardial blood flow ratios are depressed in congestive heart failure under baseline conditions, the endocardial/epicardial ratio decreases further with stress, e.g., during either adenosine or reactive hyperemia. These data suggest that one mechanism of failure of the severely hypertrophied heart may involve inadequate coronary perfusion to the subendocardium. PMID- 2970786 TI - Effects of vasodilators on the coronary circulation in congestive heart failure. AB - Pressure or volume overload of the myocardium increases the wall stress, particularly of the subendocardium, and leads to hypertrophy. Even though cardiac hypertrophy is viewed as a beneficial compensatory process that normalizes wall stress, the increased muscle mass carries with it the need of increased blood supply. Overall flow per unit mass is similar at rest in hypertrophic and normal hearts but a reduction of flow to the subendocardium and an increase in minimal coronary vascular resistance have been described. Thus, the potential exists for a vasodilator-induced steal mechanism shunting blood away from potentially ischemic areas. Angiotensin-converting enzyme inhibitors reduced myocardial oxygen consumption and coronary blood flow in parallel manner in some studies, indicating preserved coronary autoregulation, but there is also some evidence of a coronary vasodilator effect. Calcium antagonists reduce coronary vascular resistance and improve the myocardial demand-supply ratio, but the clinical usefulness of the newer compounds with supposedly little or no negative inotropic effects remains to be established. Hydralazine improved the myocardial oxygen demand-supply ratio in patients with dilated cardiomyopathy, but metabolic function may deteriorate more often after hydralazine than after angiotensin converting enzyme inhibitors in patients with coronary heart disease. Similar observations have been made using alpha-adrenergic blockers. Although progress has been made in the understanding of the coronary circulation and the influence of vasodilators in congestive heart failure, many questions await clarification using refined or new methodology. PMID- 2970787 TI - Amelioration of ischemia during angioplasty of the left anterior descending coronary artery with an autoperfusion catheter. AB - A new autoperfusion balloon angioplasty catheter with sideholes proximal and distal to the balloon--facilitating distal blood flow during inflation--was compared with standard angioplasty catheters in a prospective, randomized study with blinded data analysis. Hemodynamic and electrocardiographic markers of ischemia after 1 minute of standard or autoperfusion catheter inflations were compared with ischemia after control inflation with standard balloons. In the patient group randomized to standard balloon inflation only, ST-segment elevation after control inflation with a standard balloon catheter was 0.37 +/- 0.04 mV; ST segment elevation after final balloon inflation with a standard catheter was unchanged at 0.35 +/- 0.04 mV (difference not significant). In the group randomized to the autoperfusion catheter, control inflation with a standard catheter resulted in 0.48 +/- 0.1 mV ST elevation; final inflation with the autoperfusion catheter demonstrated 0.16 +/- 0.09 mV ST elevation (p less than 0.005). Autoperfusion catheter inflation was continued for 2 minutes without change in electrocardiographic findings: ST segments remained at 0.08 +/- 0.03 mV, unchanged from 0.07 +/- 0.03 mV before angioplasty (difference not significant). Thus, while coronary angioplasty performed with standard catheters resulted in marked ST-segment elevation, in patients undergoing angioplasty with the autoperfusion catheter, ischemia was generally not seen, despite sustained balloon inflation for 2 minutes. PMID- 2970788 TI - Clinical safety and efficacy of once-a-day amlodipine for chronic stable angina pectoris. AB - This study evaluated the safety, antianginal and antiischemic effects of amlodipine, a dihydropyridine calcium antagonist, in patients with chronic stable angina pectoris. The patients (n = 29) were evaluated during a 26-week single blind dose titration phase followed by a 6-week double-blind placebo-randomized withdrawal phase. No patient withdrawals resulted from adverse events directly related to amlodipine. A comparison of the antianginal effects of amlodipine in the single-blind phase with the placebo phase showed a reduction in anginal episodes (p less than 0.001) and a decrease in sublingual nitroglycerin usage (p less than 0.01) that persisted in the treated double-blind group (n = 12). The place-bo double-blind group (n = 10) had a reduction in anginal episodes, but no significant change in sublingual nitroglycerin usage. The antiischemic effects of amlodipine were evident; there was an increase in exercise tolerance and a reduction of ST-segment depression, as seen in the 24-hour after-dose (range 23 to 30 hours) exercise tread-mill test. During single-blind therapy, total exercise time (p less than 0.001) and time to 1 mm ST depression (p less than 0.001) displayed an overall improvement. During the double-blind phase, the treated group demonstrated an improvement in total exercise time (p = 0.01) while the placebo group had no significant change. The total amount of ST depression also differed between treated and placebo groups (1.2 +/- 0.12 vs 1.8 +/- 0.17 mm, respectively, p less than 0.01). These results lend support to the clinical safety of this medication used as once-a-day therapy. PMID- 2970789 TI - Physiologic changes during supraventricular tachycardia and release of atrial natriuretic peptide. AB - Plasma levels of atrial natriuretic peptide (ANP) increase markedly during supraventricular tachycardia (SVT). Although natriuresis associated with SVT may be secondary to the augmented secretion of ANP, whether or not physiologic changes other than natriuresis can be attributed to the release of ANP has not been determined. In the present study, plasma ANP levels in 10 patients with SVT were found to be significantly (p less than 0.05) increased, from 37 +/- 11 pg/ml (mean +/- standard error of the mean) during the control period to 160 +/- 54 pg/ml at 60 minutes after the induction of SVT. Urinary sodium excretion, although insignificant, tended to increase during the 30-minute period after SVT termination. The filtration fraction determined by the ratio of creatinine to para-aminohippurate clearance significantly increased during SVT. An increase in capillary permeability seemed to have occurred as there was a rise of hematocrit, the changes of which showed a different time course from that of the urine volume. The ratio of plasma aldosterone concentration to plasma renin activity significantly decreased during SVT. As the same effects are observed after ANP infusion, these changes were attributed to ANP activity. PMID- 2970790 TI - Electrocardiographic diagnosis of left ventricular hypertrophy in the presence of complete right bundle branch block. AB - There are few data evaluating electrocardiographic diagnosis of left ventricular (LV) hypertrophy in the presence of right bundle branch block (BBB). This study assesses the relative value of multiple electrocardiographic variables for detecting LV hypertrophy in patients with right BBB. LV hypertrophy was defined as LV mass greater than or equal to 215 g calculated from the Penn method using standard M-mode echo measurements. Sixty-two patients were evaluated (ages 41 to 94) and 35 had LV hypertrophy. None of the electrocardiographic criteria was sensitive; the best was mean QRS axis less than or equal to -30 degrees (sensitivity) 52%) and excessive negative P terminal force (sensitivity 28%). Several electrocardiographic variables were specific: Sokolow index greater than or equal to 35 mm (100%), RV5 or RV6 greater than or equal to 25 mm (96.3%), RI + SIII greater than or equal to 25 mm (92.6%) and intrinsicoid deflection greater than or equal to 0.05 seconds (88.5%). Conventional electrocardiographic criteria do not reliably detect LV hypertrophy in the presence of right BBB, but several criteria are relatively specific for LV hypertrophy. PMID- 2970791 TI - Plasma catecholamine responses to balloon angioplasty in children with coarctation of the aorta. PMID- 2970792 TI - Comparison of "host cell infiltrates" in patients with follicular lymphoma with and without spontaneous regression. AB - The "host cell infiltrates" in five patients with low-grade follicular lymphoma who had spontaneous regression without therapy were studied with the use of immunohistochemical methods applied to frozen sections. These infiltrates were compared with the "host cell infiltrates" in six patients with follicular lymphoma with progressive disease. The group with progressive disease was selected to be similar to the group with spontaneous regression in age, sex, histologic characteristics, and stage of disease. The patients with spontaneous regression had significantly more T-helper cells in the host cell infiltrate than the control patients. There were no statistically significant differences between the two groups in numbers of cytotoxic/suppressor T-cells, macrophages, Tac positive cells, Leu-7-positive cells, or proliferating cells. PMID- 2970793 TI - Hepatitis B virus infection in an institution for the mentally retarded. AB - A cross-sectional survey on the prevalence of hepatitis B serologic markers and hepatitis B virus DNA was performed in a population of 493 mentally handicapped males. Special interest was focused on age-related variables such as age at entry into the institution and on duration of residency. Furthermore, the differences with regard to the prevalence of hepatitis B markers found in Down's syndrome residents and other mentally retarded persons were analyzed. In a longitudinal study, the impact of the presence of hepatitis B virus DNA in serum was studied. Overall, 62.1 per cent of residents had serologic evidence of infection with hepatitis B virus, while 16.7 per cent of those residents with markers of infection were positive for hepatitis B surface antigen (HBsAg). Hepatitis B virus DNA was found in 24 per cent of HBsAg carriers (all positive for hepatitis B e antigen (HBeAg). In residents whose age at entry was less than 15 years, those with Down's syndrome were more often carriers of HBsAg than other mentally retarded residents. In addition, Down's syndrome residents more often had serum hepatitis B virus DNA compared with residents with other forms of mental retardation. A young age at entry was recognized as an important factor with regard to the prevalence of hepatitis B markers. From the longitudinal studies, it appeared that loss of hepatitis B virus DNA from serum indicated imminent loss of HBeAg and normalization of alanine aminotransferase values. Knowledge of the hepatitis B virus DNA status of HBsAg carriers in these institutions may therefore provide a valuable tool in attempts to reduce the transmission of this infection. PMID- 2970795 TI - Low concentrations of ANP cause pressure-dependent natriuresis in the isolated kidney. AB - The effect of alteration in renal perfusion pressure on the response of the isolated perfused rat kidney to concentrations of alpha-human atrial natriuretic peptide (ANP) within the pathophysiological range has been examined. At a perfusion pressure of 90 mmHg ANP concentrations of 50, 200, and 1,000 pmol/l were without effect on any parameter tested. At a perfusion pressure of 130 mmHg 50 pmol/l ANP produced an increase of 3.13 +/- 0.68 mumol/min in sodium excretion (UNa V), compared with a fall of 0.33 +/- 1.04 mumol/min in controls (P less than 0.02); fractional excretion of sodium (FENa) rose by 1.45 +/- 0.36% vs. -0.12 +/- 0.47% (P less than 0.05); glomerular filtration rate (GFR) was unchanged. At 200 and 1,000 pmol/l larger changes in UNa V and FENa were seen; only at 1,000 pmol/l was a significant effect on GFR observed. In contrast, frusemide (furosemide) at concentrations of 10 and 100 mumol/l was natriuretic at both 90 and 130 mmHg, with lesser absolute but greater proportional changes being seen at the lower pressure. It was concluded 1) the response of the isolated kidney to ANP is critically dependent on perfusion pressure, 2) at elevated levels of perfusion pressure the isolated kidney can respond to levels of ANP within the upper physiological and pathophysiological range. PMID- 2970794 TI - Causes of death to age 30 in Down syndrome. AB - To look at the underlying cause of death (U.C.O.D.) data for Down syndrome (DS), we studied 324 DS individuals who died out of a total of 1,337 DS births occurring in 1,066,508 consecutive live births during the years 1952-81 inclusive. U.C.O.D. rates, separated into ICD-9 classifications, for the DS population were compared with those of the age-matched general population. In general, an individual with DS is significantly more likely to die than the age matched general population over all ages studied up to age 30. The greatest absolute likelihood of dying is under 1 year, but the age group with the greatest relative risk of dying (17.2) is very definitely between ages 1-9. In order, the three categories for causes of death in DS with the greatest relative risk are congenital anomalies, circulatory system, and respiratory system. PMID- 2970796 TI - Direct vasodilatory action of atrial natriuretic factor on canine glomerular afferent arterioles. AB - Studies were performed to examine whether atrial natriuretic peptide (ANP) has a direct action on glomerular afferent arterioles, and if so, whether the action is mediated by guanosine 5'-cyclic monophosphate (cGMP). A single superficial afferent arteriole was dissected from the canine kidney and perfused with the single glomerular perfusion technique described by Osgood et al. [Am. J. Physiol. 244 (Renal Fluid Electrolyte Physiol. 13): F349-F354, 1983]. Norepinephrine (NE, 1 x 10(-6) M) significantly increased arteriolar resistance, calculated from the perfusion rate and arteriolar pressure. Synthetic human ANP (hANP) provoked afferent arteriolar dilation and attenuated the NE-induced increase in arteriolar resistance with 1 x 10(-10) to 1 x 10(-6) M concentrations. This vasodilatory effect was significantly potentiated by 2-o-propoxyphenyl-8-azapurin-6-one (M&B 22,948, 4 x 10(-12) M), a cGMP phosphodiesterase inhibitor, probably due to a sequential interaction of synergistic drugs. Also, the 1 x 10(-4) M concentration of 8-bromoguanosine 5'-cyclic monophosphate or dibutyryl guanosine, 5'-cyclic monophosphate (DBcGMP) lessened NE-induced arteriolar constriction, but DBcAMP did not. We conclude from these observations that ANP has a direct vasodilatory action on canine glomerular afferent arterioles, and that this ANP-induced vasodilation is mediated by enhanced cGMP synthesis. PMID- 2970797 TI - Hormone effects on NaCl permeability of rat inner medullary collecting duct. AB - It has been proposed that regulation of NaCl excretion occurs in part by hormonal effects on NaCl permeability in the inner medullary collecting duct (IMCD). We carried out experiments in isolated perfused terminal IMCDs to determine whether atrial natriuretic factor (ANF), vasopressin, or deoxycorticosterone (DOC) affects NaCl permeability. Apparent Cl- or Na+ permeabilities (PCl and PNa) were determined by measuring ion fluxes resulting from imposed electrochemical gradients. Transepithelial resistance (RT) was calculated from voltage deflections at the perfusion and collection ends of the tubule, which resulted from perfusion end current injection (cable analysis). ANF [rat ANF-(1-28), 100 nM in the peritubular bath] significantly decreased PCl from 2.20 to 1.84 x 10( 5) cm/s and did not alter PNa (1.11 to 1.18 x 10(-5) cm/s). ANF also decreased PCl in IMCDs from DOC-treated rats (1.14 to 0.98 x 10(-5) cm/s). Vasopressin (10 nM in the peritubular bath) did not affect PCl. RT averaged 39.3 omega.cm2 in IMCDs from control rats and was significantly increased to 62.3 omega.cm2 in tubules from DOC-treated rats. Neither ANF nor vasopressin significantly affected RT in either group. We conclude the following: 1) the results do not support the hypothesis that ANF causes natriuresis by increasing the NaCl permeability of the terminal IMCD. Instead, ANF significantly decreases the chloride permeability. 2) Vasopressin does not affect NaCl permeability. 3) Mineralocorticoid-induced antinatriuresis may be due in part to reduced NaCl permeability in the terminal IMCD. PMID- 2970798 TI - Atrial natriuretic factor does not inhibit basal or angiotensin II-stimulated proximal transport. AB - Atrial natriuretic factor (ANF) can functionally overcome the effects of angiotensin II in several tissues. Since ANF and angiotensin II in physiological concentrations have opposite effects on renal sodium excretion, we evaluated whether functional antagonism of the two hormones occurs in the proximal convoluted tubule, as has been recently reported with use of the shrinking split droplet technique. We used the more conventional technique of in vivo microperfusion to measure the response to systemic ANF (0.5 micrograms.kg-1.min 1) when the endogenous angiotensin II level and proximal transport were normal or when transport was first stimulated by systemic angiotensin II administration (20 ng.kg-1.min-1). In both cases, ANF did not significantly alter bicarbonate, chloride, or water transport in either the early or late proximal convoluted tubule. This inability by ANF to directly affect proximal transport is consonant with the known lack of high-affinity receptors and appropriate second messenger system for ANF in the proximal convoluted tubule. PMID- 2970799 TI - Renal metabolism of atrial natriuretic peptide in the rat. AB - The purpose of this study was to identify the nephron and cell sites involved in the renal metabolism of alpha-rat atrial natriuretic peptide (alpha-rANP) and to examine the degradation products of the peptide. In micro-dissected nephrons 125I labeled ANP degradation rate was highest in proximal convoluted (PCT) and straight tubules and lowest in glomeruli and papillary collecting tubules, indicating that the sites of ANP degradation and of the receptors that mediate its biological activity in the nephron do not coincide. Among subcellular fractions of cortical homogenates, the luminal membranes were the most active in metabolizing ANP. In contrast, ANP degradation by isolated basolateral membranes was negligible, and the basolateral uptake route in intact tubules did not contribute significantly to its catabolism. Cortical homogenates, luminal membranes, and isolated PCT degraded ANP without evidence of saturation up to pharmacological concentrations (10(-6) M) of the peptide. A major intermediate metabolite was rapidly formed by luminal membranes and was identified with use of a sequence and compositional analysis. This metabolite had the same amino acid sequence as ANP with a cleavage at position Cys7-Phe8, and the disulfide bridge was preserved. These results demonstrate a rapid degradation of ANP by kidney tissue and suggest that the luminal membrane of the proximal tubule is a major nephron site of ANP catabolism. PMID- 2970800 TI - Renal effects of atrial natriuretic factor are independent of dopamine1 receptors. AB - This study examined whether the renal effects of atrial natriuretic factor (ANF) are mediated by dopamine1 (DA1) receptor activation. Intravenous infusion of low dose ANF (0.0025 micrograms.kg-1.min-1) in euvolemic, pentobarbital sodium anesthetized male mongrel dogs enhanced urine flow (V) by 71 +/- 14% (mean +/- SE) and urinary sodium excretion (UNaV) by 457 +/- 172% (P less than 0.05). Renal blood flow (RBF) was unchanged. Administration of pharmacological doses of ANF (0.1 microgram.kg-1.min-1) into the renal artery in volume-expanded dogs increased RBF by 26 +/- 6, V by 56 +/- 15, and UNaV by 101 +/- 42%. The selective DA1 receptor antagonist SCH-23390 (0.5 microgram.kg-1.min-1 iv) did not affect the response to ANF at either dose. The selective DA1 agonist, fenoldopam, increased RBF by 45 +/- 3, V by 94 +/- 27, and UNaV by 61 +/- 15% in volume expanded dogs. With SCH-23390, fenoldopam increased RBF by only 16 +/- 6% whereas V and UNaV decreased by 16 +/- 10 and 17 +/- 10%, respectively. Accordingly, the failure of DA1 receptor-blocking doses of SCH-23390 to antagonize the response to ANF, at pharmacological or physiological doses, indicates that the renal effects of ANF, in the dog, are independent of DA1 receptor activation. PMID- 2970801 TI - Angiotensin II modulates the intrarenal effects of atrial natriuretic peptide. AB - The mechanism by which atrial natriuretic peptide (ANP) increases renal water and solute excretion is not fully understood. We studied the renal effects of ANP and angiotensin II (ANG II) separately and together in uninephrectomized conscious dogs (n = 7) in sodium metabolic balance (80 meq/day). Exogenous ANG II and ANP were without measurable systemic effects as demonstrated by absence of changes in blood pressure, plasma aldosterone concentration, and plasma renin activity. The quantity of ANG II that had significant renal effects that were without measurable systemic effects was 0.2 pmol.kg-1.min-1. Three infusion rates of ANP had significant renal effects (1, 10, and 20 pmol.kg-1.min-1). These quantities of ANP caused significant diuresis, natriuresis, kaliuresis, and increased glomerular filtration rate without significant changes in renal plasma flow. ANG II alone caused significant antidiuresis, antinatriuresis, and decreased glomerular filtration rate and renal plasma flow. When ANG II and ANP were given together, no change in urinary flow rate, urinary sodium or potassium excretion, or renal plasma flow was observed, whereas glomerular filtration rate increased. Filtration fraction increased significantly with ANG II and ANP separately and together. Intrarenal ANP prevents the ANG II-induced decrement in urinary sodium excretion and urine flow rate. ANP may play an important role in escape from the sodium-retaining action of intrarenal ANG II. PMID- 2970802 TI - CaBPr facilitates intracellular diffusion for Ca pumping in distal convoluted tubule. AB - The system of renal Ca transport in the rat is modeled in terms of two classes of processes: a nonsaturable flux that predominates in the proximal tubule, and an active, vitamin D-dependent flux with major expression in the distal convoluted tubule. There transport is against an electrochemical gradient, with much of the efflux probably mediated by the Ca/Mg-ATPase. Calculations of the rate of free Ca diffusion in tubular cells indicate that an unaided flux would be only one seventy-seventh of that found experimentally. It is suggested that the vitamin D induced renal calcium binding protein, CaBPr, Mr approximately 28,000, in raising total cellular calcium by three orders of magnitude, increases the transcellular Ca flux and thus the free intracellular Ca ion concentration at the basolateral pole, allowing the Ca/Mg-ATPase to function near its maximum. Analysis of the rate of nonsaturable Ca flux throughout the kidney tubule suggests a paracellular pathway via bulk flow, following water that is driven osmotically. Evaluation of whole animal data in terms of these two classes of calcium fluxes indicates that our model is consistent with experimental observations and assigns a functional role to active calcium transport. PMID- 2970803 TI - Sympathetic changes during development of cardiac hypertrophy in aortic constricted rats. AB - Cardiac hypertrophy is frequently associated with sympathetic changes that include increased myocardial norepinephrine turnover, depleted myocardial norepinephrine stores, decreased myocardial responses to sympathetic nerve stimulation, and elevated plasma catecholamines. To better understand these events mechanistically, the time course of each was assessed in rat hearts subjected to aortic constriction-induced pressure overload. There was no evidence of increased left ventricular norepinephrine turnover in abdominal aortic constricted rats, when compared with sham-constricted animals, during the first 3 days postoperatively. Moreover, their turnover rate constants tended to be lower during this period, then increased significantly by day 7. Plasma catecholamines were increased and left ventricular norepinephrine stores were decreased only on day 7. Heart rate responses to maximal sympathetic nerve stimulation were significantly reduced on the third postoperative day. Thus the decrease in norepinephrine stores coincided with changes in left ventricular norepinephrine turnover and plasma catecholamines, whereas the reduction in heart rate responses did not. This pattern suggests an initial reflex decrease in myocardial sympathetic tone, followed by baroreceptor resetting, with an eventual increase in general sympathetic outflow. The period of increasing catecholamine stimulation occurred after a relative left ventricular hypertrophy had developed but before a significant increase in absolute left ventricular mass. Thus catecholamines may still importantly contribute to the hypertrophy seen in this model. PMID- 2970804 TI - Atrial natriuretic factor-induced vasodepression occurs through central nervous system. AB - To characterize the blood pressure and heart rate effects of atrial natriuretic peptide (ANP) in the brain, we administered 20 micrograms/kg of atriopeptin III in 5 microliters of 0.9 normal saline into the fourth ventricle of awake, freely moving, spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. ANP produced a 13 +/- 1 mmHg decrease in mean arterial blood pressure (MAP) in the SHR (P less than 0.001 vs. base line or saline control, n = 10) and a 9 +/ 2 mmHg decrease in the WKY (P less than 0.02). Heart rate did not change significantly in response to ANP. To determine whether an interaction with the adrenergic nervous system played a role in the effects of ANP, we administered 100 ng yohimbine HCL, an alpha 2-antagonist, by intracerebroventricular injection, 45 min before ANP and completely prevented the ANP-induced decrease in MAP. In contrast, 100 ng intracerebroventricular prazosin, an alpha 1-adrenergic antagonist, had no significant influence on the MAP effect induced by ANP. A third group of SHR was pretreated with intracerebroventricular 6-OH dopamine to deplete central catecholamines or with saline. The rats pretreated with 6-OH dopamine (n = 6) had no significant response to ANP, which was administered 9 days later. This was significantly different from the saline-pretreated control group (n = 6), which responded with a 19 +/- 3 mmHg decrease in MAP (P less than 0.025). These studies indicate that the administration of ANP into the fourth ventricle of the brain decreases the MAP of rats through an interaction with the central alpha 2-adrenergic nervous system.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970805 TI - Reflex activation of sympathetic nervous system by ANF in humans. AB - Recent studies in experimental animal preparations suggest that ANF might alter sympathetic nervous system function. In the present investigation, direct recordings of postganglionic muscle sympathetic nerve activity were obtained from the peroneal nerve of conscious human volunteers. These data and hemodynamic parameters were recorded before and during infusions of atrial natriuretic factor (ANF, 99-126) or placebo (isotonic saline) in 10 subjects. Base-line ANF (36.5 +/ 3.8) increased to 329 +/- 22 pg/ml during 20-min infusions of ANF (15 ng.kg 1.min-1). This did not alter heart rate or blood pressure but reduced central venous pressure (CVP) by 47 +/- 10% (P less than 0.01). Base-line-integrated sympathetic activity (14.4 +/- 2.4 bursts/min) increased 30 +/- 12% during ANF infusion (P less than 0.05). However, when CVP was fixed at control levels with head-down tilt or lower body positive pressure, sympathetic activity was unchanged from pre-ANF base-line levels. These data indicate that exogenous infusions of ANF reduced CVP and unloaded cardiopulmonary baroreceptors. This elicits reflex increases of muscle sympathetic efferent activity. When CVP is maintained at control levels, ANF does not alter sympathetic neural outflow to muscles. PMID- 2970806 TI - Atrial natriuretic peptide inhibits postural release of renin and vasopressin in humans. AB - The effects of infusions of atrial natriuretic peptide (ANP) on the hormonal and hemodynamic responses to head-up tilt were investigated in six healthy adults. Head-up tilt at 45 degrees for 2 h during placebo saline infusion caused a 7% fall in blood volume accompanied by increases in plasma renin activity (PRA) and plasma arginine vasopressin (AVP) of 112 and 175%, respectively. Head-up tilt was repeated during an infusion of ANP producing a four- to sixfold increase in plasma ANP concentrations. This resulted in an 18% fall in plasma volume, yet despite this greater fall in plasma volume, PRA did not change. Two subjects experienced vasovagal symptoms toward the end of the ANP infusions accompanied by large increases in plasma AVP. In the other four subjects, plasma AVP remained unchanged during ANP infusions. Both procedures resulted in similar increases in plasma norepinephrine levels and in heart rate. Infusion of ANP prevents the posturally stimulated release of renin and AVP. PMID- 2970807 TI - Renal response of anesthetized rats to low-dose infusion of atrial natriuretic peptide. AB - Studies were performed in rats to determine the minimum infusion rate of atrial natriuretic peptide (ANP) associated with detectable changes in renal function and to determine the change in plasma levels of the peptide produced by these infusion rates. Synthetic ANP-(4-28) was administered to anesthetized euvolemic rats at rates ranging between 10 and 230 ng.kg-1.min-1 for 30 min. Significant natriuresis and diuresis were seen with an infusion of 20 ng.kg-1.min-1. At this rate of infusion, plasma ANP averaged 279 +/- 19.9 pmol/l (vs. 158 +/- 11.8 pmol/l in control rats). A transient increase in K excretion was seen with infusions higher than 100 ng.kg-1.min-1. There was no measurable change in glomerular filtration rate up to an infusion of 160 ng.kg-1.min-1. A significant decrease in mean arterial pressure was only seen with an infusion of 230 ng.kg 1.min-1. In volume-expanded rats, infusion of ANP at 10 ng.kg-1.min-1 induced a significant natriuresis. Our results indicate that natriuresis and diuresis are caused by an infusion of ANP which produces changes in plasma ANP concentration that may well result from stimulation of endogenous ANP release. In contrast, changes in K excretion, glomerular filtration rate, and arterial blood pressure may require changes in plasma ANP that are not easily achievable by physiological interventions. PMID- 2970808 TI - The immunophenotyping of extramedullary myeloid cell tumors in paraffin-embedded tissue sections. AB - Extramedullary tissue infiltrates of acute myeloid leukemia are rare and often difficult to recognize in routine paraffin-embedded tissue sections. Since appropriate therapy for these tumors depends on their precise identification, we have studied a series of tissues infiltrated with primitive myeloid cells using monoclonal and polyclonal antibodies capable of labeling cells of the myeloid/monocytic system in paraffin-embedded tissue sections. The current retrospective study involved tissues from 15 patients (eight men and seven women) with a mean age of 51 years (range, 23-77). A diagnosis of extramedullary myeloid cell tumors had been made on the basis of routine histology, chloroacetate esterase cytochemical stain, and--in some cases--electron microscopy. Paraffin embedded tissue sections were cut and stained employing the alkaline phosphatase antialkaline phosphatase (APAAP) immunocytochemical procedure with monoclonal antibodies against leukocyte-common antigen (PD7/26-2B11), restricted components of the leukocyte-common antigen (UCHL1, 4KB5), granulocytes (Mac-387, Leu-M1), leukocytes (MT1, MT2, LN1, LN2), HLA-DR (LN3), and elastase (NP57), as well as polyclonal antibodies against lactoferrin, lysozyme, alpha-1-antitrypsin, and alpha-1-antichymotrypsin. Results indicate that antibodies against Mac-387, elastase, and lysozyme are most useful in the recognition of neoplastic myeloid cells. We conclude that tissues containing granulocytic tumors can be identified in paraffin-embedded tissue sections using a panel of antibodies and the APAAP procedure. PMID- 2970809 TI - Undifferentiated primary hepatic non-Hodgkin's lymphoma in childhood. AB - This report describes a case of primary non-Hodgkin's lymphoma occurring in the liver of an 11-year-old boy. Preoperative imaging established a large mass confined to the right lobe of the liver; complete removal was effected by right hepatic lobectomy. At presentation, there was serological evidence of active hepatitis B infection, which was confirmed histologically in the nonneoplastic portion of resected liver. The right lobe of liver was virtually replaced by a multinodular tumor mass that histologically resembled small-cell lymphoma. No evidence of either a B- or T-cell lineage could be established by immunophenotyping. However, immunohistochemical staining for panleukocyte markers demonstrated membrane staining. Gene rearrangement studies were not available. The patient remains well with no evidence of disease 2 1/2 years after surgery. PMID- 2970810 TI - "Shadow" cells as clues to follicular differentiation. PMID- 2970811 TI - The in vivo effect of triethylphosphine gold (auranofin), sodium aurothiomalate and azathioprine on lymphocyte subsets of patients with rheumatoid arthritis. AB - The percentages of CD5+ (pan T), CD8+ (mainly T suppressor) and CD4+ (mainly T helper) cells were studied in 80 patients with rheumatoid arthritis (RA), grouped according to medication into four groups: 1) controls, not receiving remission inducing therapy, 2) patients on oral gold (triethylphosphine gold, auranofin), 3) patients on parenteral gold (sodium aurothiomalate) and 4) patients on azathioprine. The total number of lymphocytes was not influenced by treatment with auranofin or sodium aurothiomalate, while the number of lymphocytes was depressed in the azathioprine-treated group (P less than 0.01). Neither treatment with azathioprine or parenteral gold, nor treatment with azathioprine influenced the CD4+/CD8+ ratio. PMID- 2970812 TI - Functional units in rainbow trout (Salmo gairdneri, Richardson) liver: II. The biliary system. AB - The intrahepatic biliary system was studied in the rainbow trout (Salmo gairdneri), a teleost known to form liver neoplasms after exposure to various carcinogens. Normal adults (N = 25) were examined using light microscopic, enzyme histochemical, and transmission and scanning electron microscopic methods. In light micrographs, longitudinal arrays of hepatocytes appeared as double rows incompletely divided by elongated darkly stained cells. Electron micrographs showed tubules of five to nine pyramidally shaped hepatocytes with their apices directed toward a central biliary passageway and their bases directed toward sinusoids. Sequentially, beginning with hepatocytes, biliary passageways included canaliculi, preductules, ductules, and ducts. Canaliculi were short and joined transitional passageways (preductules) formed by junctional complexes between plasma membranes of hepatocytes and small, electron-dense cells with a high nuclear to cytoplasmic ratio. Ductules, completely lined by biliary epithelial cells, occupied central regions of hepatic tubules. Relatively elongated, ductular cells were intimately associated with surrounding hepatocytes, separated from them by only a thin extracellular space devoid of a basal lamina. Epithelium of bile ducts included cuboidal through mucus-laden columnar cells, surrounded by basal lamina and, in larger ducts, by fibroblasts, smooth muscle cells, and a capillary plexus. Bile ducts and hepatic arterioles, but not venules, were distributed together. The ultrastructure of biliary epithelium, periductular, and periductal cells is presented. PMID- 2970813 TI - Hemodynamic effects of doxacurium chloride in patients receiving oxygen sufentanil anesthesia for coronary artery bypass grafting or valve replacement. AB - Doxacurium chloride is an investigational long-acting neuromuscular blocking drug, which has been shown to be devoid of cardiovascular side effects when administered in modest doses to healthy patients. This is the first hemodynamic study of doxacurium in adult patients with cardiac disease. Forty-one patients scheduled to undergo cardiac surgery were studied. Anesthesia consisted of induction with midazolam 0.2-0.3 mg/kg and sufentanil 0.01-0.03 mg followed by an infusion of sufentanil at 0.03-0.06 mg.min-1. Baseline hemodynamic data were collected during a stable state of sufentanil anesthesia. Doxacurium was then administered in doses of 1, 2, or 3 times its ED95 of 0.025 mg/kg. Hemodynamic measurements were repeated at 2, 5, and 10 min after doxacurium injection in the absence of surgical stimulation. An additional group of control patients received saline instead of doxacurium. Baseline hemodynamic measurements were similar among groups. There was a slight decrease in heart rate in all groups over time. However, there was no significant difference between the groups of patients receiving doxacurium and the control group in which the heart rate decreased progressively from 52 beats/min at baseline to 49 beats/min 10 min after doxacurium administration. At no time was there any significant change in mean arterial pressure, right atrial pressure, or cardiac output. Likewise derived hemodynamic variables including cardiac index, stroke volume, and pulmonary vascular resistance were unchanged. In addition to the decrease in heart rate, the hemodynamic changes, which reached statistical significance, were clinically insignificant and occurred predominantly in the group of patients receiving doxacurium 0.08 mg/kg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970814 TI - Hospital resources used for inpatient and ambulatory surgery. AB - New hospital and physician payment schemes encourage physicians to participate actively in efforts to minimize hospital resource use. As an example of the type of evaluations anesthesiologists may conduct, we examined hospital resources used for comparable groups of inpatients (INPTs) and day surgery unit (DSU) patients. Although INPTs and DSU patients undergoing surgical arthroscopy of the knee or diagnostic laparoscopy were similar with regard to age, physical status, and staff surgeon, more preoperative tests were performed for INPTs than for DSU patients (P less than .05). Hospital costs for these tests were four times greater for INPTs than for DSU patients. Operating room time was from 20 to 45 min longer for INPTs than for DSU patients (P less than .05). Recovery room time was from 25 to 52 min longer for DSU patients (P less than .05). Per patient nursing labor costs paralleled operating and recovery room times. These kinds of analyses are important in identifying opportunities to improve resource use, in assessing institutional costs for surgical care, and in designing strategies that allow institutions and physicians to respond to cost containment pressures. PMID- 2970815 TI - Doppler-guided "percutaneous" radial artery cannulation in small children. PMID- 2970816 TI - Bronchoconstriction by nebulized metabisulfite solutions (SO2) and its modification by ipratropium bromide. AB - A nebulized solution of sodium metabisulfite (MBS) induced bronchoconstriction in nine subjects with asthma. An anticholinergic drug, ipratropium bromide (IPRA), in doses (500 to 700 micrograms) which were found to inhibit methacholine-induced bronchoconstriction, protected three of the nine subjects against nebulized MBS. These results suggest that both cholinergic and non-cholinergic reflex mechanisms are involved in MBS-induced bronchoconstriction. PMID- 2970817 TI - Captopril and vasculitis. PMID- 2970818 TI - [Physiopathological approach to seborrhea of the scalp]. AB - Objective assessment of seborrhea of the scalp has seldom been reported in the literature. We have used a variant of the method described by Saint-Leger and Leveque to measure the sebum excretion rate in normal scalp, male pattern alopecia, pityriasis amiantacea, pityriasis capitis and alopecia areata. The sebum excretion rate was significantly increased in male pattern alopecia, significantly decreased in pityriasis amiantacea and showed a trend towards reduction in alopecia areata. Pityriasis capitis was not associated with changes in the excretion of sebum. PMID- 2970819 TI - [Propionibacterium acnes septicemia in a mycosis fungoides]. PMID- 2970820 TI - Is abdominal wall tenderness a useful sign in the diagnosis of non-specific abdominal pain? AB - Pain arising from the abdominal wall has been implicated as a cause of non specific abdominal pain (NSAP), and the presence of abdominal wall tenderness (AWT) has been proposed as an accurate diagnostic test for NSAP. One hundred and fifty eight patients admitted to hospital with abdominal pain were tested for the presence of positive AWT. In 53 patients the final diagnosis was appendicitis and positive AWT was found in five. Thirty eight patients were found to have a variety of other recognised pathological diagnoses, none of whom had a positive AWT. In 67 patients a diagnosis of NSAP was made in the absence of other pathological diagnosis, 19 of whom had positive AWT, which was significantly different from the other diagnostic groups. This study confirms the presence of AWT in up to 28% of patients with NSAP, and suggests that testing for AWT is of value in patients with abdominal pain, although a positive AWT is not as accurate a predictor of NSAP as previously reported. PMID- 2970821 TI - Efficacy of a levamisole bolus in field infections of bovine nematodes. AB - Forty four parasite free calves were divided into eight groups, four of six and four of five calves. Two of the groups of six were given a bolus containing 22 g of levamisole in a formulation designed to release the anthelmintic for ninety days. One group of treated calves was put out to graze at the beginning of May with one of the untreated larger groups on a field contaminated with O ostertagi and C oncophora, and the other treated group grazed with controls on a field known to have the same species plus D viviparus. On mid August these resident calves were removed, slaughtered and worm counts estimated. At the same time, the other groups of untreated calves were put onto the same fields to act as tracers of the level of infection. After grazing for one month they were housed, slaughtered and worm counts carried out. The faecal egg counts of treated calves were much less than those of the controls until the end of July. Control calves on the paddock infected with lungworm were severely affected by parasitic bronchitis whereas treated calves although infected did not display clinical symptoms. In the resident calves, O ostertagi burdens were reduced by 65 and 67% and C oncophora by 97% compared to untreated controls. In the tracer calves, there was no difference in burdens of O ostertagi and D viviparus but large differences in the number of C oncophora established.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970823 TI - Effect of urea on the partial reactions and crystallization pattern of sarcoplasmic reticulum adenosine triphosphatase. AB - Steady-state ATPase activity, calcium binding, formation of phosphorylated enzyme intermediate with ATP in the presence of Ca2+, or with Pi in the absence of Ca2+, and association of ATPase molecules into bidimensional crystals, were studied using vesicular fragments of sarcoplasmic reticulum. The vesicles were exposed to increasing concentrations of urea in order to produce stepwise perturbations of protein structure and to test the effect of such perturbations on the partial reactions and crystallization pattern of sarcoplasmic reticulum ATPase. It was found that low concentrations of urea produce specific inhibition of Pi binding and enzyme phosphorylation with Pi (but not with ATP). Intermediate concentrations of urea reduce calcium binding affinity and cooperativity, while the ability of the enzyme to be phosphorylated with ATP and to form dimeric arrays is retained. These observations demonstrate that the sarcoplasmic reticulum ATPase is sensitive to physical perturbations producing specific and reversible changes in the Pi and calcium binding domains. These changes interfere with enzyme turnover, indicating that conformational effects related to binding and dissociation of Pi and calcium are tightly coupled to catalysis and energy transduction. Higher concentrations of urea produce irreversible denaturation, accompanied by total inhibition of calcium binding, enzyme phosphorylation with ATP, and association of ATPase chains in bidimensional crystals. Under these conditions, protein unfolding is manifested by a sharp reduction in the fluorescence of intrinsic tryptophan residues and of a covalently bound probe. These observations suggest that dimeric association and a tendency to form bidimensional crystals correspond to a basic property of the enzyme, which is linked to its native structure and whose character may change in the presence of ligands and/or during the catalytic cycle. On the other hand, the decavanadate induced crystallization pattern cannot be interpreted in terms of a mechanistic relationship of ATPase dimerization with one of the intermediate states of the catalytic cycle. PMID- 2970822 TI - Effect of age and housing location on antibiotic resistance of fecal coliforms from pigs in a non-antibiotic-exposed herd. AB - The relationship of age and housing location to single antibiotic resistance, multiple antibiotic resistance, and resistance patterns of fecal coliforms obtained during a 20-month period from pigs in a herd that was not exposed to antibiotics for 126 months was determined. Bacteria resistant to single and multiple antibiotics were isolated more frequently (P less than 0.01) from pigs under 7 months of age. A greater proportion of isolates from pigs over 6 months of age was sensitive to the 13 antimicrobial agents tested (P less than 0.01), while a smaller proportion showed resistance to single (P less than 0.05) and multiple (P less than 0.01) antibiotics. More than 80% of the resistant isolates were resistant to tetracycline, streptomycin, or sulfisoxazole. Resistance was greater (P less than 0.01) for pigs in the finishing unit than for those on pasture. Resistance to ampicillin, carbenicillin, and tetracycline was greater (P less than 0.05) for pigs in the finishing unit than for those in the farrowing house. More isolates from pigs on pasture were sensitive to all antimicrobial agents tested (P less than 0.01). A greater proportion of isolates from pigs in the finishing unit showed resistance to a single antibiotic (P less than 0.01). The data from this study suggest that exposure to antibiotics is not the only factor that influences the prevalence of bacteria that are resistant to single and multiple antibiotics in the feces of domestic animals and that considerable research is needed to define the factors influencing antibiotic resistance in fecal bacteria. PMID- 2970824 TI - Phosphofructokinase is responsible for the fructose 2,6-bisphosphate inhibition of hexokinase in tissue extracts. AB - Mammalian and yeast hexokinases were reported to be reversibly inhibited by fructose 2,6-bisphosphate in the presence of cytosolic proteins (H. Niemeyer, C. Cerpa, and E. Rabajille (1987) Arch. Biochem. Biophys. 257, 17-26). Reinvestigation of this finding using a radioassay with [14C]glucose as substrate showed no effect of fructose 2,6-bisphosphate on hexokinase activity of rat liver cytosols. Detailed reexamination of the spectrophotometric assay resulted in the observation that the fructose 2,6-bisphosphate-dependent inhibition was a function of the cytosolic phosphoglucose isomerase and phosphofructokinase activities compared to the amount of glucose-6-phosphate dehydrogenase used as auxiliary enzyme. The diminution or loss of the fructose 2,6-bisphosphate dependent inhibition produced in aged cytosols was restored by addition of crystalline muscle phosphofructokinase, as well as by decreasing the amount of glucose-6-phosphate dehydrogenase in the assay. When phosphoglucose isomerase, phosphofructokinase, and hexokinase activities were separated by DEAE chromatography of liver cytosol, no fructose 2,6-bisphosphate-dependent inhibition of hexokinase was found in any single fraction of the chromatogram. However, combination of fractions containing both phosphoglucose isomerase and phosphofructokinase displayed the fructose 2,6-bisphosphate-dependent inhibition on either endogenous hexokinase or added yeast hexokinase. From these results we conclude that the activation of phosphofructokinase elicited by fructose 2,6 bisphosphate is responsible for the hexokinase inhibition observed in the coupled spectrophotometric assay. PMID- 2970825 TI - [Neocarzinostatin therapy of advanced renal cell carcinoma]. AB - Fifteen patients with advanced renal cell carcinoma underwent chemotherapy of neocarzinostatin (NCS). First, 6-8 mg of NCS was infused into renal artery at angiography; second, 6-8 mg was infused by the same route just before ligation of renal artery at nephrectomy, if possible; and third, 2 mg was given intravenously at 2-week intervals, about 10 times. One patient who suffered from metastatic renal cancer on rs. tibia was treated by femoral arterial injection of 2 mg NCS 5 times. By treatment of NCS for advanced renal cell carcinoma 2 out of fifteen patients achieved complete regression, one patient showed partial regression and four a minor response. Effective rate (CR + PR) of NCS for metastatic renal cancer was 20%, and the response rate (CR + PR + MR) was 47%. We consider that NCS is presently the most effective drug for renal cell carcinoma. PMID- 2970826 TI - Purpura fulminans and transient protein C and S deficiency. PMID- 2970827 TI - Prenatal screening for Down's syndrome. PMID- 2970828 TI - Maternal and fetal beta endorphin: effects of pregnancy and labour. PMID- 2970829 TI - [Does the handicapped child still need a pediatrician?]. PMID- 2970830 TI - Leukocyte common antigen expression in lymphomas. PMID- 2970831 TI - [Changes in hemostasis of dental importance. Diagnostic system]. PMID- 2970833 TI - [Myocardial reaction of the right ventricle to lung resection]. AB - Right-sided pulmonectomy (resection of 63-65% of the lung parenchyma) in white noninbred rats resulted in development of chronic cor pulmonale, that develops according to the stages: I--from the time of the operation up to the 10th-15th days after the operation--the stage of acute disturbances and mobilization forces of the organism; II--from the 11th-15th up to the 90th day is the stage of a relative steady compensatory hypertrophy of the cardiac right ventricle; III- after the 90th day--the stage of decompensation. The hypertrophy of the right ventricle myocardium transfers into its dilatation. Amount of cardiomyocytes and their nuclei in 1 mg of the right ventricle tissue progressively decreases, quantity of multinuclear cardiomyocytes increases, ploidy of the nuclei changes: number of tetraploid nuclei decreases, octaploid nuclei appear. Lethality among the animals is 56%. PMID- 2970832 TI - Effect of anabolic steroids, alone or in combination with antibiotics, on bull performance, carcass traits and meat quality characteristics. AB - Two trials were conducted with 32 and 39 finishing Belgian white-red bulls, treated with anabolic agents during 55 to 60 days prior to slaughter. Xenobiotic androgens and estrogens, together with (Exp. I) or without progesterone (Exp. II) were injected at three-week intervals. Diets consisted of maize silage and 7.5 g concentrate daily per kg liveweight. Half of the animals within each experiment received an antibiotic for the entire fattening period lasting about 285 days. Lasalocid-sodium (Exp. I) and virginiamycin (Exp. II) were incorporated at 65 mg per kg concentrate. Anabolic agents improved average daily gain from 1.06 to 1.20 kg (P less than 0.10) and from 1.24 to 1.42 kg (P less than 0.05) respectively. In both trials steroidal growth promoters stimulated intake and improved feed efficiency (P less than 0.10). Feed antibiotics did not significantly affect daily gain during this short term finishing period. A reduced feed intake (P less than 0.05) and improved feed conversion were observed for lasalocid, while there was no difference for virginiamycin. Dry matter intake amounted to 80.5 and 76.3 g per kg metabolic weight and 9.02 and 8.41 kg per kg gain, respectively for control and lasalocid treated animals. The data were 79.6 and 80.3 g and 7.30 and 7.34 kg, respectively, for the experiment with virginiamycin. No significant interaction between anabolics and antibiotics was obtained with respect to gain and feed efficiency. Dressing percentage, carcass composition, carcass grading and meat traits were not affected by anabolic treatment or antibiotic supplementation. PMID- 2970834 TI - [Histochemical characteristics of the muscle fibers of the biceps and triceps brachii muscles in human ontogeny]. AB - By means of morphometrical and histochemical methods for revealing myosin ATPase and SDG activity development of various types of muscle fibers (MF) has been studied in the postmortem material, using m. biceps and m. triceps brachii in human ontogenesis. The flexors and extensors have features in common in the dynamics of the MF maturation, and some distinctive peculiarities. The appearance of histochemical distinctions between the MF takes place on the 5th-6th months of the intrauterine development. Morphofunctional specialization begins with formation of tonic fibers. During the 1st-2nd years phasic fibers form. A relative amount of fast MF in both muscles increases at the age of 11-12 years. The dynamics of final specialization of the MF is connected with stages of sexual maturation. The first stage of the sexual maturation (about 14 years of age) is connected with decrease in the relative amount of the MF of glycolytic type of energy supply and corresponding increase in the number of oxidative type structures. From 15-17 years of age a final differentiation begins, it is connected with an intensive transversal growth of all the MF and distinguish of thick glycolytic MF. The m. biceps brachii has a relatively greater amount of oxidative fibers, and the m. triceps brachii, glycolytic ones. The transversal section area of the MF in the m. triceps brachii exceeds that of the m. biceps brachii, beginning from the 7th month of the intrauterine development up to 14 years of age. The investigation performed does not reveal any anticipating development either in the flexors or in the extensors. The differentiating processes in the m. biceps and m. triceps brachii occur nearly simultaneously. PMID- 2970835 TI - [The structure of iatrogenic diseases based on autopsy data from Moscow and Leningrad]. AB - The occurrence and pattern of disorders attributed to diagnosis and treatment were assessed comparatively by autopsy findings obtained in Moscow and Leningrad. The analysis suggested high frequency rate for iatrogenic diseases promoted by surgical interventions and chemotherapy. Some of these can be traced to physician's errors while delivering therapeutic and diagnostic procedures. The authors cover methodological approaches to gaining information, point to a high informative value of autopsy records, argue that iatrogenic diagnosis must be reflected in autopsy protocols and registered all over the country. PMID- 2970836 TI - [Complications of x-ray endovascular recanalization and their role in thanatogenesis]. AB - In line with clinical benefits observed in the diagnosis and treatment of arteriopathies, invasive methods currently introduced in a wide practice entail some negative sequelae presenting with specific complications with occasional lethal outcomes. Because the complications are so closely related to pathogenesis and outcomes of the disease, they should be thoroughly analyzed in view of their role and place in the conceptions of thanatogenesis and pathological diagnosis in which they can be introduced as the main, concurrent or competitive disease, registered as complications depending on the role in the thanatogenesis and initial status of the patient assessed both by the clinician and the pathologist before the invasive procedure. PMID- 2970837 TI - Spectrum of cerebrospinal fluid findings in various stages of human immunodeficiency virus infection. AB - This report summarizes the results of neurologic and cerebrospinal fluid (CSF) study findings in over 400 of the 649 human immunodeficiency virus-infected US Air Force personnel, evaluated as of Dec 31, 1987. Eighty percent of these patients were entirely asymptomatic and immunologically normal, 13% had low T helper lymphocyte counts and/or cutaneous anergy, and only 7% had opportunistic infection. Sixty-three percent of all patients had some CSF abnormality. Sixty percent of the asymptomatic group had at least one abnormal result, over 25% had three or four CSF abnormalities, and over 7% had five or six abnormal values. When patients with evidence of blood-brain barrier leak were excluded, significant differences were seen between disease groups with regard to CSF glucose, CSF IgG levels, and CSF IgG synthesis. No human immunodeficiency virus related central nervous system abnormalities were found on neurologic examination in immunologically intact asymptomatic patients regardless of CSF findings. No clear-cut predictor of impending central nervous system complications has, as yet, been identified from the CSF parameters studied. PMID- 2970838 TI - Laparoscopy ... a useful technique for the general surgeon. AB - A series of 227 consecutive laparoscopies performed over 3 years at a provincial hospital in the North Solomons Province is presented. There were no deaths due to the procedure and morbidity was 2.4%. Positive findings were revealed in 89% of patients, allowing rapid decisions to be made regarding patient management: a vital consideration in a busy surgical unit with limited resources. Laparoscopy provides an efficient, safe, rapid and direct approach to many abdominal problems encountered in general surgery, particularly liver disease, blunt abdominal trauma, intraperitoneal infection and neoplasia. PMID- 2970839 TI - Coronary thrombolysis with and without nifedipine in pigs. AB - To investigate whether addition of Ca2+ antagonists adds to the beneficial effects of thrombolysis we studied recovery of regional myocardial performance in pigs, in which occlusive thrombi were induced by electrical stimulation, with and without addition of nifedipine to the thrombolytic agent. To this end, four different groups of animals with thrombotic coronary occlusion were studied. Groups 1 and 2 received either saline or intracoronary nifedipine (0.1 micrograms.kg-1.min-1) 15 min after coronary artery occlusion. Groups 3 and 4 were treated with the thrombolytic agent plasmin which was infused directly into the left anterior descending coronary artery (LADCA) at a rate of 2 U.min-1. The animals in group 4 also received intracoronary nifedipine. 4 h after thrombus formation the animals were sacrificed. No important differences in systemic hemodynamics were observed between the four groups of animals. Reperfusion occurred only in the animals which received plasmin, with or without nifedipine. After intracoronary plasmin regional blood flow increased from 7 +/- 2 to 40 +/- 7 ml.min-1.100 g-1 in the LADCA-perfused subepicardial and from 9 +/- 2 to 30 +/- 6 ml.min-1.100 g-1 in the LADCA-perfused subendocardial layers. The combination of plasmin and nifedipine increased flow to the LADCA-perfused subepicardial layers from 8 +/- 2 to 74 +/- 21 ml.min-1.100 g-1 and that to the subendocardial layers from 8 +/- 2 to 57 +/- 16 ml.min-1.100 g-1 (in both cases: p less than 0.05 vs. plasmin alone). However, addition of nifedipine did not enhance recovery of regional myocardial function or high-energy phosphate metabolism. Because reperfusion was accompanied by a high ventricular ectopic activity, the question may be raised of whether reperfusion of ischemic myocardium which does not result in functional recovery could be deleterious. PMID- 2970840 TI - Propranolol and thyroxine-induced hypertrophic rabbit hearts: effect on heart size and regional O2 supply/consumption variables. AB - The purpose of this study was to determine the effect of acute and chronic propranolol on heart size and regional O2 supply/consumption variables in thyroxine (T4)-treated rabbit hearts. New Zealand white rabbits were given 0.5 mg/kg T4 for 3 or 16 days with and without concomitant 2 mg/kg propranolol. Another group was given 16 days of propranolol alone and another 3-day T4 group was given 2 mg/kg propranolol 1 h before the experiment began. Another group served as control. Myocardial blood flows were determined using radioactive microspheres and small arteriolar and venous O2 saturations were determined using microspectrophotometry. Treatment with T4 for 3 or 16 days increased the heart weight/body weight ratio, myocardial blood flow, and regional O2 consumption. 16 day T4 treatment resulted in myocardial flow 195% and O2 consumption 300% above control group values. When propranolol was given chronically along with T4, heart weight/body weight ratios did not increase to the degree seen with 3 or 16 days of T4, alone. Propranolol given acutely in 3-day T4-treated animals, resulted in a reduced O2 consumption and O2 extraction, though not to the extent seen with chronic propranolol treatment of T4-treated animals. Acute propranolol treatment slightly reduced myocardial blood flow in 3-day T4-treated animals, while chronic treatment significantly reduced it. Chronic propranolol treatment in 16-day T4 treated animals resulted in a significant reduction in flow and O2 consumption. Thus, T4 treatment increased O2 consumption, flow, and heart size and these effects could be attenuated using acute and chronic propranolol. PMID- 2970841 TI - Ventricular myosin pattern of spontaneously hypertensive turkeys is unaffected by labetalol treatment. AB - In most animal species, left ventricular hypertrophy due to pressure overload is associated with an advantageous increase of the "slow" V3 isomyosin. In contrast, in spontaneously hypertensive turkeys, the development of left ventricular hypertrophy is associated with the synthesis of a "fast" V1-like isomyosin, with high incidence of cardiac failure. This could be related to the high catecholamine levels found in these animals. This is why we studied the ventricular myosin pattern after lowering of blood pressure and regression of cardiac hypertrophy obtained by means of labetalol, and alpha- and beta-blocking drug which inhibits the effects of catecholamines. From the 2nd to the 32nd week of age, 22 turkeys were treated with increasing doses of p.o. labetalol (from 20 to 35 mg/kg body weight daily) and 16 other turkeys were given daily p.o. placebo. Blood pressure and heart rate were periodically measured by an indirect method. After sacrifice, the degree of cardiac hypertrophy was evaluated by the biventricular weight to body weight ratio, ventricular myosin was purified, Ca++ activated ATPase activity assessed, and ventricular myosin pattern was determined by two-dimensional gel electrophoresis of myosin heavy chains. Plasma and cardiac catecholamines were measured by high performance liquid chromatography. Throughout the study period, blood pressure and heart rate were significantly reduced in the labetalol-treated animals as compared to the untreated ones. At the end of the study period, the ventricular mass was significantly lower in the labetalol group. Nevertheless, no differences were observed in ventricular myosin pattern and Ca++-activated ATPase activity levels between the two groups. In the labetalol group, an increase in plasma catecholamines and only a slight, but not significant, increase in cardiac catecholamines was found. These data indicate that in spontaneously hypertensive turkeys, the synthesis of the "fast" V1-like isomyosin is not influenced by known pathophysiological stimuli like blood pressure, cardiac hypertrophy and catecholamines. PMID- 2970842 TI - Changes in ECG, plasma and myocardial lipids in experimental myocardial hypertrophy in rats. AB - ECG, systolic blood pressure (BP), the ratio (R) of grams of myocardial mass/100 g of body mass, total lipids, cholesterol, triglycerides and phospholipids in blood plasma and the left ventricular myocardium, as well as the plasma free fatty acids, were investigated in 58 male Wistar rats 3, 30 and 180 days after operation, in a model of myocardial hypertrophy (MH) induced by experimental coarctation hypertension, after the method of Selye. An attempt was made to correlate some functional and metabolic indices which characterize the development of this type of MH. On a background of progressively rising BP and parallel increasing R, ECG changes were recorded. They were typical of the respective stage of arterial hypertension and MH and expressed mostly in a shifting of the electrical axis of the heart to the left and in essential repolarization disturbances. The most significant changes in the studied lipid fractions were found 30 days after the induction of hypertension. The pathological changes manifested on the 180th day are discussed in relation to age, the stage of hypertension and especially in relation to the developing hypoxic and ischaemic myocardial damage. PMID- 2970843 TI - Analysis of the phosphofructokinase subunits and isoenzymes in human tissues. AB - The 6-phosphofructo-1-kinase (PFK) subunits and isoenzymes were studied in human muscle, heart, brain, liver, platelets, fibroblasts, erythrocytes, placenta and umbilical cord. In each tissue, the subunit types in the native isoenzymes were characterized by immunological titration with subunit-specific antibodies and by column chromatography on QAE (quaternary aminoethyl)-Sephadex. Further, the subunits of the partially purified native isoenzymes were resolved by SDS/polyacrylamide-gel electrophoresis, identified by immunoblotting, and quantified by scanning gel densitometry of silver-stained gels and immunoblots. Depending on the type of tissue, one to three subunits were detected. The Mr values of the L, M and C subunits regardless of tissue were 76,700 +/- 1400, 82,500 +/- 1640 and 86,500 +/- 1620. Of the tissues studied, only the muscle PFK isoenzymes exhibited one subunit, which was the M-type subunit. Of the other tissues studied, the PFK isoenzymes contained various amounts of all three subunits. Considering the properties of the native PFK isoenzymes, it is clear that, in human tissues, they are not simply various combinations of two or three homotetrameric isoenzymes, but complex mixtures of homotetramers and heterotetramers. The kinetic/regulatory properties of the various isoenzyme pools were found to be dependent on subunit composition. PMID- 2970844 TI - A 36 kDa monomeric protein and its complex with a 10 kDa protein both isolated from bovine aorta are calpactin-like proteins that differ in their Ca2+-dependent calmodulin-binding and actin-severing properties. AB - Interaction of plasma membrane with the cytoskeleton involves a large number of proteins, among them a 36 kDa protein that was found to be involved in the interaction with actin filaments. We have isolated a 36 kDa protein from bovine aorta as a monomer and in a complex with a 10 kDa protein. Partial amino acid sequence determinations show that the 36 kDa and 10 kDa proteins isolated from bovine aorta are analogous to or identical with corresponding proteins purified from bovine intestine already described by Kristensen, Saris, Hunter, Hicks, Noonan, Glenney & Tack [(1986) Biochemistry 25, 4497-4503]. We report here that the association of the 10 kDa protein with the 36 kDa protein confers specific calmodulin-binding and actin-severing properties on the complex that are not possessed by the 36 kDa monomer alone. These findings suggest that the protein complex could be involved in thin-filament-related structures or could modulate some Ca2+-regulated events mediated by calmodulin. PMID- 2970845 TI - Subcellular localization of the sulphation reaction of heparan sulphate synthesis and transport of the proteoglycan to the cell surface in rat liver. AB - We report on the incorporation of radiolabelled sulphate into proteoglycan in the 'in situ'-perfused rat liver. After 5 min virtually all of the [35S]sulphate was incorporated into heparan sulphate; no partially sulphated precursors were detected. Pulse-chase experiments, followed by centrifugation in gradients of sucrose and metrizamide, showed that, at 5 min, the heparan sulphate was associated predominantly with the Golgi membranes. Over the next 20 min, intact proteoglycan appeared at the plasma membrane. At intermediate times the heparan sulphate was detected simultaneously in two distinct populations of membrane vesicles. Whether the heparan sulphate in these two populations has two different destinies (e.g. plasma membrane or secretion) is not yet clear. Subfractionation of the Golgi membranes showed that the N-sulphotransferase co-purified with the heparan [35S]sulphate and was separable from the galactosyltransferase of glycoprotein synthesis, confirming that the Golgi membrane system is functionally segregated. Subfractionation also permitted an almost 100-fold purification of the N-sulphotransferase over the homogenate: this will provide an excellent starting material for isolation and further characterization of the enzyme. PMID- 2970846 TI - Identification of beta-N-acetylhexosaminidase A in mouse tissues with the fluorigenic substrate 4-methylumbelliferyl-beta-N-acetylglucosamine 6-sulphate. AB - beta-N-Acetylhexosaminidase from mouse tissue was separated into its constituent isoenzymes on DEAE-cellulose and its activity was monitored with 4 methylumbelliferyl-beta-N-acetylglucosamine and 4-methylumbelliferyl-beta-N acetylglucosamine 6-sulphate. Forms corresponding to the human isoenzymes A (acidic), B (basic) and an 'intermediate' form were present in mouse liver and spleen, whereas in kidney the B and 'intermediate' forms predominated, with A present only as a minor component. In brain the 'intermediate', A and C activities were detected. Testis had predominantly A activity, whereas epididymis, the tissue with the highest specific activity of beta-N acetylhexosaminidase, had an abundance of the 'intermediate' form, but was almost entirely lacking in the A form. PMID- 2970847 TI - Reversal of caldesmon function by anti-caldesmon antibodies confirms its role in the calcium regulation of vascular smooth muscle thin filaments. AB - Direct evidence that caldesmon is the Ca2+-regulated inhibitory component of native smooth muscle thin filaments is provided by studies using caldesmon specific antibodies as antagonists. The antibodies reverse caldesmon inhibition of actomyosin ATPase and abolish Ca2+-regulation of native aorta thin filament activation of myosin ATPase. This effect is a result of antibody binding to the caldesmon on the filament thereby inactivating it and not due to antibody-induced caldesmon dissociation from the filament. The antibodies, however, neutralise caldesmon only in systems using skeletal muscle myosin and not in those using smooth muscle myosin; this implies that smooth muscle myosin prevents appropriate antibody binding to caldesmon perhaps because smooth muscle myosin binds to caldesmon thus preventing access of antibody to antigenic sites. PMID- 2970848 TI - NMR relaxation measurements detect four intermediate states of ATPase and transport cycle of sarcoplasmic reticulum Ca2+-ATPase. AB - At least four of the intermediate states of Ca2+-ATPase (and presumably ion transport) can be trapped and characterized using water proton relaxation measurements. Gd3+ binds to two occluded Ca2+ transport sites on Ca2+-ATPase which have a low accessibility to solvent water. In the presence of the MgATP analogue Co(NH3)4AMPPCP, a new state for bound Gd3+ with one less water of hydration) is observed. In the presence of Co(NH3)4ATP or ATP, two additional states for bound Gd3+ are detected by NMR, the first of which probably represents an intermediate state of ATP hydrolysis. The latter is the most occluded Gd3+ site yet observed in these studies and corresponds to the highly occluded E1-P state observed with CrATP (Vilsen and Andersen, Biochim. Biophys. Acta 898, 313 (1987). PMID- 2970849 TI - A specific insulin receptor and tyrosine kinase activity in the membranes of Neurospora crassa. AB - Cells of the wall-less ("slime") strain of Neurospora crassa possess specific high affinity insulin binding sites on their cell surface. 125I-labeled bound insulin was not displaced from these cells by insulin-like growth factor II (IGF II), and was only weakly displaced by IGF-I and proinsulin. Cross-linking of 125I labeled insulin with N. crassa cells using disuccinimidyl suberate resulted in the labeling of a single band of ca. 67 kDa m.w. on a polyacrylamide gel. Two proteins of ca. 66 and 59 kDa m.w. were purified from detergent solubilized plasma membrane preparations by passage over an insulin-agarose affinity matrix. Antibodies against an autophosphorylation site on the human and Drosophila insulin receptors (anti P2) immunoprecipitated a single phosphoprotein of ca. 50 kDa m.w. from detergent solubilized plasma membranes, which possessed protein tyrosine kinase activity when histone H2 was used as substrate. PMID- 2970850 TI - [Percutaneous penetration of mucopolysaccharide-polysulfate ester from a combination preparation]. AB - The cutaneous penetration of mucopolysaccharide polysulfate (MPS) from a commercial available combination drug (Mobilat) could be shown by means of histochemical and immunohistological methods. Deposition of MPS from ointment and gel in corium and subcutis of different animal species has been improved by the keratolytic activity of the salicylic acid component. There are differences in the intensity of absorption of MPS not only in the various animal species, but also caused by the particular galenic preparations investigated. PMID- 2970851 TI - Alteration of atrial natriuretic peptide binding sites in spontaneously hypertensive rats. AB - Atrial natriuretic peptide (ANP) binding sites were studied by quantitative autoradiography in young (4-week-old) and adult (14-week-old) spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) control rats. Young SHRs had fewer ANP binding sites in the smooth muscle layer of thoracic aorta. Both young and adult SHR had undetectable ANP binding in superior cervical and stellate sympathetic ganglia, pituitary gland, thymus and spleen, and lower binding in subfornical organ and choroid plexus. Conversely, in young SHRs and WKY rats, ANP binding site density was similar in adrenal zona glomerulosa, kidney glomeruli, and papilla. The SHRs have notable tissue specific decrease in ANP binding site concentration. This alteration may relate to the pathophysiology of genetic hypertension. PMID- 2970852 TI - Long-term humoral and hemodynamic effects of celiprolol. AB - To evaluate the humoral and hemodynamic (both systemic and renal) effects of chronic treatment with celiprolol, six out-patients with mild to moderate uncomplicated essential hypertension received placebo for 1 month and celiprolol (400 mg qid) for 6 months. At the end of placebo and of the first and sixth month of treatment, blood pressure (BP), heart rate (HR), renal plasma flow (RPF), glomerular filtration rate (GFR), plasma renin activity (PRA), aldosterone (ALD) and noradrenaline (NA), urinary enzymes (NAG: N-acetyl-beta glucosaminidase, AAP: alanine aminopeptidase) were measured. Compared to placebo, celiprolol significantly and steadily reduced BP and HR. However, although the systemic hemodynamic effect was constant during the whole period of the study, the reduction of renovascular resistance and of plasma noradrenaline, detectable at the first month of therapy, disappeared at the sixth month. However, PRA, plasma aldosterone, GFR, and urinary enzymes did not change. These findings suggest that the antihypertensive effect of celiprolol is well maintained over the 6-month period; the drug did not exert any adverse effect on the kidney, and chronic celiprolol treatment does not influence renal hemodynamics. PMID- 2970853 TI - Isradipine (PN 200-110) versus hydrochlorothiazide in mild to moderate hypertension. A multicenter study. AB - The effects of 10 weeks of treatment with isradipine (ISRP), a new dihydropyridine Ca antagonist, was evaluated in a prospective, randomized, double blind, parallel group, hydrochlorothiazide (HCTZ) controlled study in patients with mild to moderate hypertension. Of 98 patients enrolled, 73 completed the study and were deemed valid for efficacy analyses; 36 in the ISRP group and 37 in the HCTZ group. Monotherapy with ISRP significantly (P less than 0.001) decreased (mean +/- SD) sitting systolic blood pressure (BP) from 146 +/- 11 mm Hg to 128 +/- 11 mm Hg and diastolic BP from 100 +/- 4 mm Hg to 83 +/- 5 mm Hg. Heart rate during the plateau period was not significantly different (76 +/- 11 vs 78 +/- 11 bpm) between the ISRP and HCTZ groups. These reductions in BP were comparable to monotherapy with HCTZ. The mean reduction in diastolic BP with ISRP (17 +/- 6 mm Hg) was significantly (P less than 0.05) greater than that with HCTZ (14 +/- 5 mm Hg). The mean doses for ISRP and HCTZ were 12 mg/day and 60 mg/day, respectively. There was no significant difference in frequency of common side effects (headache, nausea, fatigue, dizziness, palpitations) between the two groups. However, transient or intermittent peripheral edema occurred more frequently in ISRP group. Four patients in ISRP group (two due to edema and two due to palpitations) and two patients in HCTZ group (due to poor BP control) were discontinued from the study. Our results indicate that ISRP in doses of 5 to 10 mg bid is as effective as HCTZ as monotherapy in the treatment of mild to moderate hypertension. PMID- 2970854 TI - Pharmacokinetics of single and consecutive doses of cilazapril and its depressor effects in patients with essential hypertension. AB - The pharmacokinetic properties and antihypertensive effects of cilazapril, a long acting angiotensin-converting enzyme (ACE) inhibitor, were investigated in five patients with mild to moderate essential hypertension (mean age 57 years, mean serum creatinine 1.2 mg/dL, mean glomerular filtration rate 69 mL/min/1.73m2, mean blood pressure 158/94 mm Hg). All patients were hospitalized and placed on a constant sodium diet (7 g of NaCl/day) throughout the study. After an overnight fast, a 1.25-mg dose of cilazapril was given orally once a day for 5 or 8 days. On the first and last days of treatment, blood samples were taken and blood pressure was measured. All patients tolerated cilazapril with no untoward effects. Cilazapril induced a significant decrease in both systolic and diastolic blood pressure, and its antihypertensive effect was still present 24 hours after administration. Serum ACE activity was markedly suppressed for at least 24 hours. The peak plasma concentrations (Cmax) of cilazapril and its diacid were 117 and 24.6 ng/mL on the first treatment day, and 144 and 31.1 ng/mL on the last day. The area under the plasma concentration time curve (AUC) of cilazapril and its diacid were 408 and 227 ng.h/mL on the first day, and 501 and 305 ng.h/mL on the last day. In looking at the data gathered on the first and last treatment days, no significant differences were noted in Cmax and AUC values. These results suggest that cilazapril has a long-lasting effect and is a useful antihypertensive agent in controlling blood pressure in patients with mild to moderate essential hypertension. PMID- 2970856 TI - Efficacy and safety of cilazapril, a new angiotensin-converting enzyme inhibitor. AB - Cilazapril (CIL), a new angiotensin-converting enzyme inhibitor, was evaluated for 16 weeks in 29 patients with mild to moderate essential hypertension (diastolic pressure 95 mm Hg to 115 mm Hg). Twenty-four patients (83%) normalized their blood pressure (BP) (diastolic pressure less than 90 mm Hg), 11 with low dose CIL, six with high-dose CIL, one with high-dose CIL plus low-dose thiazide, and six with high-dose CIL and high-dose thiazide. Three withdrew because of side effects (fatigue, bloating, and polyuria). Statistically significant reductions in sitting and standing systolic and diastolic pressures occurred at 8 and 16 weeks on CIL. There was no change in standing or sitting heart rate, white blood cell count, creatinine clearance, urine protein levels. This is the first long term data on this new converting enzyme inhibitor in human beings. PMID- 2970855 TI - Effects of fosenopril, a once-daily angiotensin-converting enzyme inhibitor, on resting and exercise-induced changes of blood pressure, hormonal variables, and plasma potassium in essential hypertension. AB - Fosenopril, a new angiotensin-converting enzyme (ACE) inhibitor, is a prodrug that is converted to its active diacid metabolite after intestinal absorption. Its excretion is equally divided between hepatic and renal routes. This study details the efficacy of fosenopril at rest and during exercise on blood pressure, plasma potassium, plasma renin activity, and plasma aldosterone in 11 hypertensive males, mean age 55 years. Individual subjects underwent an identical exercise protocol (bicycle ergometry) on placebo and on active treatment. Supine mean blood pressure fell from 116 to 100 mm Hg, P less than 0.005, and at 9 minutes of exercise mean blood pressure fell from 137 to 125 mm Hg, P less than 0.01 on fosenopril. Plasma potassium fell slightly at rest from 4.27 to 3.96 mmol/L and during exercise from 5.23 to 4.93 mmol/L, both P less than 0.025. Plasma renin activity (ng/mL/hr) rose on fosenopril, at rest 0.94 to 4.72, and during exercise 2.06 to 10.39, both P less than 0.005. Aldosterone changes on treatment were marginal and nonsignificant. The compound was well tolerated and free of subjective and routine laboratory side effects. Its antihypertensive action is broadly similar to the other ACE inhibitors currently available. PMID- 2970857 TI - Prevention of hypertension is associated with reduced susceptibility to histamine induced arrhythmias in SHR. AB - Two groups of spontaneously hypertensive rats (SHR) were treated with alpha methyldopa (2.5 g/L in drinking water) for 12-17 weeks. One group was treated during the normal time course for development of hypertension and myocardial hypertrophy (beginning at age 4 weeks), while the other was treated after stabilization of hypertension/hypertrophy (21 weeks). Appropriate age-matched controls (WKY strain) also were treated. Intracellular microelectrodes were used to monitor action potential configuration, automaticity, and the incidence of arrhythmias (including delayed afterdepolarizations and repetitive tachyarrhythmias) in isolated left ventricles from each group. The younger rats did not develop hypertension of left ventricular hypertrophy; susceptibility of isolated left ventricles to histamine-induced (10(-5) M) arrhythmogenesis was significantly reduced. The older rats showed decreased blood pressure, although not to normal levels; further left ventricular hypertrophy was prevented, though the existing enlargement did not regress. These hearts were as susceptible to the development of histamine-induced arrhythmias as were hearts of untreated SHR. Enhanced susceptibility to arrhythmias in hypertrophied hearts reflects a preventable alteration of the myocardial cell membrane that occurs during the course of blood pressure elevation. This change may be associated with defective sarcolemmal calcium transport in hypertrophied myocardium. PMID- 2970858 TI - Attitudes of primary school children toward the physical appearance and labels associated with Down syndrome. AB - Attitudes of 80 kindergarten and 80 third-grade subjects responding to videotape excerpts of children either with or without Down syndrome and with or without the label and description "mentally retarded" were examined and affect, belief endorsements, and behavioral intentions were assessed. Multivariate analyses of variance indicated that subjects were significantly less positive in their affective evaluations and belief endorsements toward target children labeled and described as mentally retarded. Third-grade children were significantly more negative in their affective responses toward children with Down syndrome, and kindergarten children were more positive in their evaluations of such children. Possible reasons for the lack of significant gender and contact differences were discussed. PMID- 2970859 TI - Operant conditioning of vocalization rate of infants with Down syndrome. AB - Reinforcement control over vocalization rates of infants with Down syndrome was evaluated; elicitation of vocalizations by social stimulation was controlled. Parents of three infants (2.7, 5, and 8.2 months old) provided social reinforcement under two schedules: continuous reinforcement for vocalization (CRF) and differential reinforcement of other-than-vocalization (DRO). All infants produced systematically higher vocalization rates during CRF, even though the amount of social stimulation during DRO was equal to or greater than the amount provided during CRF. The results replicated findings with normally developing infants and support a contingent-stimulation model of intervention for infant vocal communication. PMID- 2970860 TI - Mean length of utterance of children with Down syndrome. AB - Mean length of utterance (MLU) of children with Down syndrome was examined and found to correlate highly with chronological age, r = .87, p less than .001, despite the children's language delays. The correlation was reliable at least up to MLU 3.00. In the 1.00 to 3.50 range, MLU also predicted grammatical complexity. PMID- 2970861 TI - Development of adaptive behavior in adolescents and young adults with autism and Down syndrome. AB - Vineland Adaptive Behavior Scales were used to assess adolescents and young adults with Down syndrome or autism. Matched for verbal mental age (MA), the groups did not differ in adaptive behavior; however, older individuals with Down syndrome had more skills than did younger ones in all areas measured. Skills did not change for the autistic group. For persons with Down syndrome, adaptive skills kept pace with verbal and nonverbal MA, whereas those with autism were delayed in communication and socialization relative to their nonverbal MA. PMID- 2970862 TI - Weekly low-dose doxorubicin with or without high-dose medroxyprogesterone acetate as secondary treatment in metastatic breast cancer--a randomized trial. PMID- 2970863 TI - The plasma membrane calcium transporting systems in the regulation of cell calcium. PMID- 2970864 TI - Ultrasound lumbar canal measurement in hospital employees with back pain. AB - The oblique parasagittal diameter of the lumbar spinal canal at the L5-S1 level was measured in 49 employees of the Wm Jennings Bryan Dorn Veterans' Hospital using real time ultrasound in a case-control study. Individuals with a canal diameter of less than 14 mm represented the lowest 10th percentile in this population and being in the narrowest 10th percentile constituted a risk factor for time missed from work because of low back pain (odds ratio 10.7). Whereas numbers in this pilot study are small, results are consistent with earlier ultrasound studies done in the United Kingdom and with other research showing increased morbidity from low back pain in individuals with small lumbar canals. Ultrasound has advantages over other modalities for measuring the size of the lumbar canal and may be useful as a preplacement screening examination in industry. PMID- 2970865 TI - A study of the structure of human complement component factor H by Fourier transform infrared spectroscopy and secondary structure averaging methods. AB - Fourier transform infrared spectroscopy was used to investigate the secondary structure of human complement component factor H in H2O and 2H2O buffers. The spectra show a broad amide I band which after second-derivative calculations is shown to be composed of three components at 1645, 1663, and 1685 cm-1 in H2O and at 1638, 1661, and 1680 cm-1 in 2H2O. The frequencies of these components are consistent with the existence of an extensive antiparallel beta-strand secondary structure. The exchange properties of the amide protons of factor H as measured in 2H2O buffers are rapid and lead to an estimate of NH proton nonexchange that is comparable with those for small globular proteins. Human factor H is constructed from a linear sequence of 20 short consensus repeats with a mean of 61 residues in each one. To investigate the secondary structure further, secondary structure predictions were carried out on the basis of an alignment scheme for 101 sequences for these repeats as found in human factor H and 12 other proteins. These predictions were averaged in order to improve the reliability of the calculations. Both the Robson and the Chou-Fasman methods indicate significant beta-structural contents. Residues 21-51 in the 61-residue repeat show a clear prediction of four strands of beta-structure and four beta turns. A structural model based on antiparallel beta-strands in the secondary structure is proposed and discussed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970866 TI - Cross-linking of three heavy-chain domains of myosin adenosinetriphosphatase with a trifunctional alkylating reagent. AB - The chemotherapeutic alkylating reagent tris(2-chloroethyl)amine (TCEA) was used as a trifunctional cross-linking reagent with a cross-linking span of 5 A for myosin subfragment 1 (S-1). When S-1 was incubated with TCEA, all three domains of 20, 26, and 50 kDa in the S-1 heavy chain were cross-linked via the highly reactive sulfhydryl group SH1 (Cys-707) on the 20-kDa domain. The cross-linking was accelerated by nucleotides. The present observation is consistent with the proposal that SH1 is close to both the 26- and 50-kDa domains of S-1 and that movement within S-1 associated with the nucleotide binding occurs around SH1 as well as around another reactive thiol, SH2 & Wong, A. G. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 6392-6396; Hiratsuka, T. (1987) Biochemistry 26, 3168-3173]. PMID- 2970867 TI - Liver composition and lipid metabolism in NZB/W F1 female mice fed dehydroisoandrosterone. AB - The beneficial effects obtained with dehydroisoandrosterone (DHA) feeding in the treatment of murine systemic lupus erythematosus are similar to those obtained with caloric restriction or with dietary manipulation of essential fatty acid availability. In this study, the fatty acid composition of selected tissues was examined in NZB/W F1 mice fed a diet containing 0.4% DHA. The effect of the DHA diet on liver composition and the activity of key hepatic enzymes involved in fatty acid synthesis and glucose metabolism was also investigated. The content of the essential fatty acid, arachidonate, was decreased in plasma cholesteryl esters and liver and kidney phospholipids in mice fed the DHA diet, yet no significant decrease in arachidonate content was observed in plasma phospholipid. The most striking change in both plasma and liver phospholipid was an increase in palmitic acid and a decrease in stearic acid, which could result from a decreased ability for fatty acid elongation. The liver mass was dramatically increased in the mice fed DHA, primarily from parenchymal cell hypertrophy, and contained little lipid. Significant changes in the activities of malic enzyme, glucose-6 phosphate dehydrogenase and pyruvate kinase, similar to those changes which occur with fasting, were observed during the initial adaptation to the DHA diet. The pyruvate kinase activity remained low, suggesting a decrease in liver glycolysis. These results are consistent with the concept that diets containing DHA result in an altered metabolism with a decreased dependence on carbohydrate metabolism and an increased metabolism of lipids. PMID- 2970868 TI - Hormonal regulation of renal function during development. AB - The present review summarizes current and new knowledge concerning the major hormonal systems that directly or indirectly affect renal function during development. The role of the renin-angiotensin-aldosterone system in regulating renal function during fetal and postnatal life is reviewed. A summary of the role of this system during fetal and postnatal stresses is also provided. The physiological role of the renal kallikrein-kinin system in the control of renal blood flow, renin release and sodium excretion during development is examined. Possible influences of the prostaglandin system on regulation of renal function and renin secretion during fetal and postnatal maturation are explored. The effect of vasopressin on the ability of the fetal and postnatal kidney to concentrate urine and regulate body fluid homeostasis is reviewed in detail. The physiologic action of vasotocin on renal sodium and water homeostasis is described. New information regarding the role of the sympathetic system in the regulation of renal hemodynamics and in the control of renal function during development is presented. Finally, recent studies demonstrating the effect of atrial natriuretic factor and corticosteroids on the developing kidney are discussed. PMID- 2970869 TI - [Immunologically mediated induction of hyperlipoproteinemia]. AB - The relationship of immune and metabolic processes, particularly, those of lipid metabolism, was studied in mice with diet induced hyperlipoproteinemia, leukemia related immune deficiency and in situ formation of antigen-antibody complexes after immunostimulation. A marked immunosuppression was observed in mice on a high-cholesterol diet, but when immune complex formation was induced after antigen injection, a high level of nonesterified cholesterol and low density lipoproteins was obtained, whereas an increase in high density lipoproteins in leukemic mice was observed. It is concluded that there exists a certain mechanism of immunologically mediated induction of hyperlipoproteinemia, which leads to the inhibition of immune response by immunoregulatory lipoproteins under conditions of antigen stimulation. PMID- 2970870 TI - [Effect of different fractions obtained by gel filtration of a chalone-containing preparation from Ehrlich ascites tumor on mitotic activity and DNA synthesis in this tumor]. AB - The effect of various fractions of chalone-containing preparation from ascite Ehrlich's tumour obtained by gel filtration on ultragel Ac-A-44 on mitotic activity and DNA synthesis in the tumour has been studied. The chalone-containing preparation (alcohol precipitate) was shown to suppress entering of tumour cells into M- and S-phase and DNA synthesis. After gel filtration, the partial division of active chalone component which inhibits entering of cells into M- and S-phase took place. The component inhibiting DNA synthesis eluted with G1-chalone. PMID- 2970871 TI - Serum levels of CD8 antigen in childhood lymphoid malignancies: a possible indicator of increased suppressor cell activity in poor-risk patients. AB - Serum concentrations of CD8 antigen were measured at diagnosis with an enzyme linked immunoassay in children with acute lymphoblastic leukemia (n = 344) or non Hodgkin's lymphoma (n = 65). All patients had detectable levels of the serum antigen, which in its soluble nonreduced form appeared to be a 52-Kd homodimer as compared with the 66-Kd surface membrane component on most thymocytes and on a subset of functionally distinct T cells (suppressor/cytotoxic). Increased serum levels of CD8 in leukemia patients were significantly related to recognized high risk prognostic features: high leukocyte count, large liver and spleen size, high serum lactic dehydrogenase level, T-cell immunophenotype, presence of a mediastinal mass, pseudodiploid karyotype, DNA index less than 1.16, and chromosomal translocation. Children with serum CD8 levels greater than or equal to 450 U/mL were more likely to fail treatment than were those with lower levels (P = .002), even in the group with non-T-cell leukemia (P = .003). In a multivariate analysis, serum CD8 antigen contributed independent prognostic information beyond that conveyed by age, leukocyte count, and race (P = .02). High serum CD8 antigen levels also correlated with advanced stages of disease in children with non-Hodgkin's lymphoma or B-cell leukemia. Children with higher serum CD8 antigen levels (greater than or equal to 700 U/mL) had a poorer treatment outcome (P = .003), even after results were adjusted for disease stage and serum lactic dehydrogenase level (P = .05). Measurement of serum levels of CD8 antigen not only has important prognostic value in childhood lymphoid malignancies but also could be useful in assessing the immunoregulatory role of T cells in patients with cancer. PMID- 2970872 TI - Activated idiotype-reactive cells in suppressor/cytotoxic subpopulations of monoclonal gammopathies: correlation with diagnosis and disease status. AB - The phenotypic pattern of peripheral blood T (PBT) lymphocytes was correlated with diagnosis and clinical status in 63 patients with monoclonal gammopathies (MGs). The numbers of lymphocytes expressing activation and CD11 determinants were significantly increased in suppressor/cytotoxic and helper/inducer subpopulations of patients with multiple myeloma (MM) and MG of undetermined significance (MGUS). The number of activated suppressor/cytotoxic cells was closely correlated with diagnosis and disease status. These cells were significantly higher in MM at diagnosis (160 +/- 88) than MGUS patients (61 +/- 79; P less than .01). Their number decreased to MGUS levels in MM in stable remission (58 +/- 53), but not in MM with tumor progression (172 +/- 102; P less than .001). In individual patients, part of these cells specifically adhered to dishes precoated with the related M-protein. No monoclonal T-beta gene rearrangement was detected in PBT and cytotoxic/suppressor subpopulations from two patients with a large proportion of activated cells. PMID- 2970873 TI - c-fms expression is a molecular marker of human acute myeloid leukemias. AB - The c-fms protooncogene product was identified as the CSF-1 or M-CSF receptor, a polypeptide growth factor that plays a major role in myelomonocytic differentiation. This led us to look for expression of c-fms in fresh acute myeloid leukemia (AML) cells, using Northern blot analysis. c-fms expression was found in the leukemic cells of 28 AML patients, regardless of their stage of differentiation, which was assessed in the French-American-British (FAB) classification. However, the level of c-fms expression was especially high in AML of the M5 stage. High levels of expression were not accompanied by either amplification or rearrangements of the c-fms gene in AML cell DNAs. In contrast, c-fms expression was not found in acute lymphoid leukemias, whether of T or B origin. Thus, c-fms expression appears as a specific molecular marker of leukemogenesis in the myeloid lineage. PMID- 2970874 TI - Stability of antigens on leukocytes in banked platelet concentrates: decline in HLA-DR antigen expression and mixed lymphocyte culture stimulating capacity following storage. AB - Repeatedly transfused thrombocytopenic patients frequently form antibodies directed against human leukocyte antigens (HLA) and become unresponsive to random donor platelet transfusions. Although exposure to foreign antigens borne on donor leukocytes appears necessary to provoke primary sensitization, the stability of leukocyte antigens during routine platelet storage is largely unknown. Accordingly, we serially measured the expression of surface markers on leukocytes derived from platelet concentrates during storage using immunofluorescence and flow cytometry. Our results indicate that the expression of class I HLA antigens, Leu-4 (T cell), and HLe-1 (pan leukocyte) remained stable on lymphocytes under standard platelet storage conditions, but that the percentage of lymphocytes bearing class II HLA antigens declined significantly over time. This decline in lymphocyte HLA class II expression was associated with a significantly diminished ability of stored leukocytes to stimulate blastogenesis in mixed lymphocyte culture. However, leukocytes retained the ability to respond in mixed lymphocyte culture (MLC) following storage. We also performed studies on lymphocytes cultured in the presence of cyclohexamide, which suggested that the expression of class I HLA antigens and B2 microglobulin are highly sensitive to the inhibition of protein synthesis, whereas the expression of class II HLA antigens, Leu-4, and HLe-1 are not. Our results may prove useful in understanding the mechanisms that lead to platelet refractoriness and in designing strategies to prevent HLA alloimmunization. PMID- 2970875 TI - Effects of undernutrition during suckling and of post-training beta-endorphin administration on avoidance performances of adult rats. AB - 1. The effects of undernutrition during suckling and of post-training beta endorphin administration on avoidance task were investigated in adult rats. 2. Young rats were undernourished from delivery until weaning (21 days) by feeding their mothers a diet containing 8% protein (w/w). Mothers of well-nourished rats were fed a 20% protein diet. After weaning, both groups of rats were fed a 20% protein diet until 90-120 days of age, when they were subjected to behavioral sessions. 3. Acquisition was measured in training sessions and retention in test sessions 24 h after training. Beta-endorphin or saline (control) was injected ip immediately after training. Rats were subjected to shuttle and step-down inhibitory avoidance sessions using footshock of 0.2 or 0.8 mA intensity. 4. Undernutrition during suckling caused hyperreactivity to 0.2 mA footshocks. Beta endorphin caused amnesia to shuttle avoidance task only in normal rats trained with 0.8 mA footshocks. In the step-down inhibitory avoidance task, beta endorphin was amnesic only for normal rats and only for 0.2 mA footshocks. Beta endorphin was not amnesic in undernourished rats. PMID- 2970876 TI - Removal and release of recirculating atrial natriuretic peptide (ANP) by rabbit lungs perfused in situ. AB - The finding that radioiodinated, synthetic ANP (125I-ANP), analogous to the ANP secreted by the right atrium, binds to specific sites of endothelial cells, smooth muscle, adrenals and lungs (1-3) raised the importance of analyzing lung ANP interactions. Rabbit lungs were accordingly loaded with 125I-ANP, 2000 Ci/mM (2.5 pM) in Krebs-albumin recirculating perfusion medium at a rate of 20 ml/min, over 27 min. 125I-ANP binding was estimated by taking samples at 2-min intervals from the perfusate. Fourteen min after recirculation, uptake was maximal (66.4 +/ 5.2%, N = 8) (X +/- SEM) and not different from that observed after a single passage through the lungs and was dose-dependent, antagonized by unlabelled ANP. The system was then changed to single-pass washout and efflux curves were obtained, collecting the entire efflux every 6 s per sample, over 15 min. Efflux curves after 125I-ANP loading fitted a bi-exponential decay model (Y(to) = Y(t)e b11t + Y(t)e-b2t). Lung ANP uptake/release may be involved in regulating systemic concentrations and hence renal and other actions of ANP. PMID- 2970877 TI - Effects of autologous mesothelial cell seeding on prostacyclin production within Dacron arterial prostheses. AB - Canine abdominal aortas have been replaced with Dacron arterial prostheses to assess the effects of mesothelial cell seeding on graft prostacyclin and thromboxane A2 release. At both 2 weeks and 6 weeks after surgery, three seeded and two unseeded control grafts were examined for prostacyclin release. In addition, thromboxane release was assessed in one seeded and one unseeded graft. Sections of aorta and graft were removed and incubated in PBS containing either 10 microM calcium ionophore A23187 or 20 microM arachidonic acid. The incubation mixture was sub-sampled at 5 min intervals over a 20 min period to assess the progressive release of prostacyclin and thromboxane A2 using a radioimmunoassay for 6-keto-prostaglandin F1 alpha and thromboxane B2 respectively. In seeded grafts, 6-keto-prostaglandin F1 alpha release averaged 15 per cent compared with aorta at 2 weeks and 45 per cent compared with aorta at 6 weeks. By contrast, release from unseeded grafts was undetectable at 2 weeks; however, by 6 weeks there was some release amounting to 15 per cent compared with aorta. There was a statistically significant increase in the release of 6-keto-prostaglandin F1 alpha from mesothelial cell seeded grafts at 6 weeks compared with unseeded grafts (P less than 0.01). Thromboxane release from the graft sections was variable and unrelated to whether the grafts had been seeded or not. These preliminary results, showing that grafts seeded with autologous peritoneal mesothelial cells release more prostacyclin than unseeded grafts, further highlight the role of the mesothelial cell as an alternative to the endothelial cell for improving the patency of arterial Dacron prostheses in the early postoperative days. PMID- 2970878 TI - An operation for incisional hernia. PMID- 2970879 TI - Immunisation of neonates at high risk of hepatitis B in England and Wales: national surveillance. AB - The results of a voluntary programme of immunisation against hepatitis B in neonates at high risk (mother being positive for hepatitis B surface antigen and without hepatitis B e antibody or having had acute hepatitis B late in pregnancy) are reported. The programme was offered in England and Wales from November 1982. Passive immunisation alone was available in the first six months of life until 1985, after which infants received passive and active immunisation from birth; in addition, some infants received passive immunisation for six months followed by a course of hepatitis B vaccine. All but a few infants received the first immunising dose within 48 hours after birth. Blood samples for analysing markers of hepatitis B virus were available at 1 year from 147 of the 223 infants given passive immunisation, 54 of the 72 given passive followed by active immunisation, and 102 of the 155 given passive and active immunisation at birth. At 1 year 11 of the 127 (9%) infants given four or more doses of specific hepatitis B immunoglobulin were positive for hepatitis B surface antigen compared with four of the 20 given three or fewer doses; 11 had levels of hepatitis B surface antibody greater than 50 IU/l. Only one of the 54 infants given passive then active immunisation was positive for hepatitis B surface antigen at 1 year and four infants had low (less than or equal to 50 IU/l) levels of hepatitis B surface antibody. Four of the 102 infants who received passive and active immunisation at birth were positive for hepatitis B surface antigen. Two had received the fill course of vaccine, whereas in the other two vaccination was incomplete or unstated. In 79 of the 89 infants who received a complete course of vaccination the level of hepatitis B surface antibody was known, and 70 had levels at 1 year greater than 100 IU/1. Reactions to immunisation were not severe at any age. The incidence of side effects was 8% for the immunoglobulin, 11% for the vaccine, and 9% when immunoglobulin and vaccine were given together. Wider collaboration in the programme is requested. PMID- 2970880 TI - Sleep disruption in Down's syndrome. PMID- 2970881 TI - Correlated asymmetries in striatal D1 and D2 binding: relationship to apomorphine induced rotation. AB - Long-Evans derived rats were tested for nocturnal, amphetamine-induced and apomorphine-induced rotation (circling behavior); the rats' left and right striata were subsequently dissected and D1 and D2 receptor densities (Bmax) were assayed in the same striatal homogenates using [3H]SCH-23390 and [3H]N methylspiperone, respectively. D1 and D2 Bmax values were correlated (r = 0.68). Moreover, left-right asymmetries in D1 and D2 Bmax values were more highly correlated (r = 0.84). Although asymmetries in D1 and D2 binding were not by themselves related to rotational behavior, an asymmetry in the ratio or balance of D1 and D2 binding was associated with the direction of apomorphine-induced rotation: the D1/D2 ratio of Bmax values was significantly higher in the striatum ipsilateral to the preferred direction of apomorphine-induced rotation. These results suggest that normal variations in numbers of D1 and D2 receptors are determined by a common mechanism, that D1 and D2 receptors are functionally coupled, and that, with respect to activation of striatal receptors, D1 is inhibitory and D2 is excitatory. The effects of apomorphine, a mixed D1 and D2 agonist, appear to reflect the balance between D1 and D2 receptors. PMID- 2970882 TI - Remote infarction from coronary artery dissection following percutaneous transluminal coronary angioplasty. PMID- 2970884 TI - The clinical application of percutaneous transluminal coronary angioplasty. PMID- 2970883 TI - Desmoid tumor: a review of eleven cases. PMID- 2970886 TI - Interaction of amiloride and hydrochlorothiazide with atrial natriuretic factor in the medullary collecting duct. AB - Medullary collecting duct function was studied using the in vivo microcatheterization technique in three groups of rats receiving amiloride, hydrochlorothiazide, or both diuretics. In each group of animals, atrial natriuretic factor (ANF99-126) was given in the second phase of the experiment. The combination of amiloride and hydrochlorothiazide resulted in a more marked natriuresis than either diuretic given as a single agent. Sodium reabsorption in the medullary collecting duct, as a fraction of the delivered load, was reduced from 64% (amiloride) and 69% (hydrochlorothiazide) to 29% (amiloride and hydrochlorothiazide). Atrial natriuretic factor reduced collecting duct sodium reabsorption when added to amiloride or hydrochlorothiazide to 23% and to 41%, respectively, but had no additional effect when given with amiloride and hydrochlorothiazide. Potassium excretion with amiloride and hydrochlorothiazide was intermediate between amiloride or hydrochlorothiazide given as single agents. With the diuretic combination, potassium transport showed no significant reabsorption or secretion along the medullary collecting duct, amiloride was associated with potassium reabsorption, and hydrochlorothiazide was associated with potassium secretion in the duct. The results confirm the importance of the medullary collecting duct as a site of diuretic action. The known additive effects of amiloride and hydrochlorothiazide on sodium excretion and the opposing effects of these agents on potassium excretion occur, to a major degree, in the medullary collecting duct. Furthermore, the additive effects of amiloride and ANF indicate that blocking of amiloride-sensitive sodium channels is not the only mechanism of action of ANF on duct salt transport in vivo. PMID- 2970885 TI - Renal actions of atrial natriuretic peptide on the postischemic kidney. AB - The objective of this study was to evaluate the renal actions of atrial natriuretic peptide (ANP) in the unilateral postischemic kidney of anesthetized dogs with a severe reduction in glomerular filtration rate. The dose of atrial natriuretic peptide (50 ng.kg-1.min-1) we gave did not alter the mean systemic arterial pressure, renal blood flow, and glomerular filtration rate in the normal kidney, as determined in foregoing studies. ANP was infused into the intrarenal artery continuously for 60 min after the release from 45 min of complete renal artery occlusion. In the vehicle-infused group, the glomerular filtration rate fell dramatically (6% of control), the renal blood flow decreased (60% of control), and the mean systemic arterial pressure tended to increase (136% of control). The urine flow rate and urinary excretion of sodium decreased significantly (25 and 25%, respectively) at 30 min after reflow in the postischemic period. Continuous renal artery infusion of ANP resulted in a marked increase in urine flow rate (246% of control) and the urinary excretion of sodium (286% of control). The administration of ANP led to an improvement in renal blood flow (99% of control) and glomerular filtration rate (40% of control), and attenuated the rise in mean systemic arterial pressure (109% of control), compared with findings in the vehicle-infused group. Plasma renin activity and prostaglandin E2 concentration in the renal venous blood were elevated after the release from complete renal artery occlusion in both groups. These results indicate that the vascular effects of ANP on the postischemic kidney were enhanced and that the peptide maintained the natriuretic effect. PMID- 2970888 TI - Intraoperative Doppler flow studies: emphasis on colour mapping. AB - Colour Doppler flow imaging (CD) is a well-established method of assessing valvular and congenital disease of the heart. The technique has proven useful intraoperatively in patients undergoing valve repair or replacement or intracardiac reconstruction. In 21 patients with various cardiac anomalies, colour Doppler flow imaging was used intraoperatively, before cardiac cannulation and again after cardiopulmonary bypass. Previously undisclosed features were demonstrated preoperatively in two patients and important residual defects after cardiopulmonary bypass in three others. Adequacy of valve replacement was confirmed in four patients and small leaks in ventricular septal defect patches were shown in another three. The procedure facilitated repair of complex cardiac fistulas in two patients. There were no apparent complications from CD. As intracardiac repairs become more complex, the role of intraoperative CD will expand. PMID- 2970889 TI - Neuroblastoma during acute lymphoblastic leukemia in remission. AB - Neuroblastoma was diagnosed in a child after a 20-month remission of a pre-B acute lymphoblastic leukemia (ALL). Clumps of atypical cells suggestive of neuroblastoma were seen in the bone marrow. They were positive for monoclonal antibody (MoAb) UJ13A (neuroblastoma cells) and negative for MoAb T29/33 (anti leucocyte common antigen CD45) with immunocytochemical staining. A right paravertebral mass displacing the kidney was demonstrated by abdominal echotomography, and serum vanilmandelic acid was slightly increased. Despite specific chemotherapy against neuroblastoma and after a transient clinical improvement, the patient died 7 months later of disseminated disease. Immunocytochemical staining on cells frozen at diagnosis of leukemia with MoAb UJ13A and T29/33 was unable to demonstrate neuroblastoma cells and showed the pattern usually observed in leukemia (UJ13A- and T29/33+). PMID- 2970890 TI - Correlation between human cell growth response to interleukin 1 and receptor binding. AB - Human recombinant interleukin 1 alpha (IL-1 alpha) and IL-1 beta inhibited the replication of the mammary tumor cell line, MDA-MB-415; stimulated division in the colon carcinoma, SW-48; and had no effect on the growth of the milk mammary line, HBL-100. Inhibition of growth was reflected in a significant decrease in DNA synthesis accompanying a transient increase in RNA synthesis. Specific binding of 125I-labeled recombinant IL-1 beta by MDA-MB-415 and SW-48 reached a maximum by 2 h of incubation, and an equivalent amount was bound by each cell type. Binding was inhibited in a dose-dependent manner by unlabeled IL-1 alpha or IL-1 beta. Scatchard plot analysis revealed that MDA-MB-415 cells expressed approximately 700 binding sites with an apparent dissociation constant of 8.8 x 10(-10) M. Reversibility of growth inhibition was independent of dose or time of incubation, but DNA synthesis did not return to control values. Flow cytometric analysis of DNA content showed that growth inhibition was cell cycle phase nonspecific with a slight reduction in the proportion of cells in S phase. The major conclusion from these studies was that inhibition or stimulation of malignant cell growth by IL-1 was related to the presence of receptor sites. PMID- 2970891 TI - Suppressor cells in the effector phase of autologous cytotoxic reactions in cancer patients. AB - Cytotoxicity was induced in lymphocytes (CL) from 10 out of 15 patients by autologous mixed lymphocyte tumor cell culture and further cultivation with recombinant interleukin-2. In cells from 3 of the 10 patients, cytotoxicity was suppressed by more than 50% when autologous peripheral blood mononuclear cells (PBMC) from the patients with large tumors were added to the autologous killing system. The cells responsible for suppressing the cytotoxicity in the effector phase were adherent or nonadherent to plastic depending on the patient examined. The T cell fraction from 1 patient significantly suppressed the cytotoxic activity, and this suppression was seen only in the autologous system. On the other hand, plastic adherent cells but not T cells from PBMC of 2 subjects suppressed the cytotoxic activity of CL. The reason why the main cell population suppressing the CL activity differed among the patients is unclear. However, the findings that the suppression was mostly abrogated following resection of the tumor mass suggested that suppressor cells, either of macrophage lineage or T cells, are induced in patients with a large tumor mass. This speculation is supported by the finding that the PBMC from a patient with tumor recurrence regained the suppressive activity. PMID- 2970892 TI - Clinical and angiographic follow-up after coronary recanalization during acute myocardial infarction. AB - Coronary angiography followed by percutaneous transluminal coronary angioplasty and/or intracoronary streptokinase infusion was performed in 50 patients 288 +/- 162 min after the onset of symptoms of acute myocardial infarction. Subocclusion of the infarct-related vessel was found in 5 patients, all of whom had angioplasty of the residual stenosis. Recanalization was achieved in 37 patients (success rate 82%). There was no procedure-related death. One patient died 4 days after the intervention. Control coronary angiography 5 +/- 2 months after the procedure in 35 of 42 patients with recanalization documented recurrence of stenosis or reocclusion in 8 (23%). Comparison of preintervention and control angiograms in 33 patients showed an increase in left ventricular ejection fraction from 55 +/- 8 to 61 +/- 13%, p less than 0.001, in patients with collaterals to the infarct-related vessel and/or recanalization within 180 min after the onset of pain, and from 55 +/- 9 to 59 +/- 8%, nonsignificant, in patients with recanalization later than 180 min and without collaterals. At follow-up 7 +/- 4 months after the procedure, 1 patient had died and 36 (86%) were asymptomatic. Good long-term results can be achieved at a reasonable risk by coronary angioplasty with or without thrombolysis in evolving myocardial infarction. Left ventricular function is better preserved in patients with collaterals and/or early recanalization. PMID- 2970893 TI - Late coronary spasm complicating successful angioplasty of the left main coronary artery. AB - A patient with successful percutaneous transluminal coronary angioplasty of the left main coronary artery is described. Two days later coronary angiography was performed because of recurrence of angina pectoris. Spontaneous spasm at the angioplasty site was documented. This potentially lethal complication issues a severe warning against angioplasty of the left main trunk. PMID- 2970895 TI - The force-velocity relationship for microtubule sliding in demembranated sperm flagella of the sea urchin. AB - We studied the relationship between the force and velocity of microtubule sliding in demembranated sperm flagella of the sea urchin, Hemicentrotus pulcherrimus, under auxotonic conditions following a quick release of the tension between sliding microtubules. The shape of the force-velocity curve was independent of the concentration of Mg-ATP over the range of 3.7 to 350 microM and appeared either linear or was the reverse of the hyperbolic curve seen for muscle. The power, calculated as the product of velocity and force, passed through a peak at c. 0.7 Fmax (the maximal isometric force). Thus, the maximal power is attained at a larger relative load than in muscle. The sliding velocity at 0.1 Fmax showed a hyperbolic dependence on Mg-ATP concentration, with a Km of 210 microM and a Vmax of 19 micron.sec-1. The maximal force did not significantly change over the Mg ATP concentration range of 3.7 to 350 microM. These results are discussed in terms of a crossbridge model similar to the one originally proposed by Huxley. It is suggested that the dynein crossbridge cycle is characterized by a relatively rapid rate of attachment and a relatively slow rate of detachment. PMID- 2970894 TI - Coexpression of opsin- and VIP-like-immunoreactivity in CSF-contacting neurons of the avian brain. AB - Cerebrospinal fluid-contacting (CSF) cells in both the septal and the tuberal areas in the brain of the ring dove are labeled by RET-P1, a monoclonal antibody to opsin that reacts with inner and outer segment membranes of rod photoreceptors in a variety of vertebrates. Immunoblot analysis of proteins from diverse brain regions, however, revealed bands of anti-RET-P1 immunoreactivity that did not correspond to opsin. Binding of RET-P1 to opsin-containing membranes, was not inhibited by membranes rich in muscarinic and beta-adrenergic receptor proteins (red blood cells, heart, lung) taken from doves. RET-P1-immunoreactive CSF contacting cells emit a dendritic process that penetrates the ependyma and ends in a knob-like terminal suspended in the ventricle. These cells also possess other processes that penetrate more or less deeply into the neuropil. Additionally, a band of labeled fibers occurs in the external layer of the median eminence. A double-label technique demonstrated that RET-P1-positive cells coexpress VIP-like immunoreactivity. VIP-positive cells in other brain areas are not RET-P1-positive. PMID- 2970896 TI - Synthesis of active center-directed peptide inhibitors of plasmin. PMID- 2970897 TI - Basal serum interferon-gamma production in hay fever patients. PMID- 2970898 TI - Inhibition of mannosidase in hybridomas yields monoclonal antibodies with greater capacity for carbohydrate labeling. AB - Labeling an antibody site specifically through its carbohydrate residues preserves more of its antigen-binding activity than does labeling through protein moieties. To boost the amount of immunoglobulin G carbohydrate capable of being labeled, we treated hybridoma cells with a mannosidase inhibitor, deoxymannojirimycin (dMM). Polyacrylamide gel electrophoresis showed formation of a glycoprotein with high mannose content, in that endo-beta-N acetylglucosaminidase H (EC 3.2.1.96) could digest the antibody from the dMM treated cells, but not from control cultures. Carbohydrate analysis confirmed this conclusion, indicating that the antibody from the dMM-treated cells had twice as much mannose as did the control antibody. The glucosamine content of the treated-cells' antibodies was half that of the control, and no additional carbohydrate residues were detectable in the antibodies secreted by the dMM treated cells. We conjugated both the dMM and control antibodies through their carbohydrate to a chelator. In labeling, the dMM antibody conjugate incorporated approximately threefold as much 111In isotope as the control conjugate. The two labeled antibodies were injected into mice and showed similar organ distributions. PMID- 2970899 TI - Automated method for phosphofructokinase inhibition test with a centrifugal analyzer. PMID- 2970901 TI - Lymphocyte subpopulations in Graves' disease. PMID- 2970900 TI - Plasma and cyst fluid levels of delta 5 and delta 4 steroid hormones in women with gross cystic breast disease. AB - delta 5 and delta 4 steroid levels were studied in the plasma and cyst fluid of women with gross cystic breast disease (GCBD). In luteal phase a significant increase in plasma levels (mean +/- SEM) of DHA (11.2 +/- 2.4 ng/ml), DHAS (1.45 +/- 0.6 micrograms/ml) and cortisol 277 +/- 15.7 ng/ml) was found; in follicular phase the mean levels were 4.09 +/- 0.47 ng/ml for DHA, 0.65 +/- 0.08 microgram/ml for DHAS and 190 +/- 46.3 micrograms/ml for cortisol. The DHA/DHAS and cortisol/androstenedione ratios were significantly higher in the plasma and lower in the cyst fluid of GCBD patients, than in the plasma of controls; the androstenedione/DHA ratio was higher in the cyst fluid than in the plasma of controls. The hormonal situation of the GCBD patients thus differed from that of the controls both in the plasma and cyst fluid, particularly as regards the delta 5 steroids. PMID- 2970902 TI - The histological effects on the guinea pig external ear of several constituents of commonly used aural preparations. AB - Medicament contact dermatitis can be troublesome in external ear disease. In this work the effects on the guinea pig external ear of instillation twice daily for 3 weeks of benzalkonium chloride, Cerumol, gentamicin, the pharmaceutical base for Gentisone HC, hydrocortisone and propylene glycol were studied using light microscopy, thickness measurements of tympanic membrane and meatal epidermis, and the mitotic index of meatal epidermis. Benzalkonium chloride induced severe inflammatory changes and markedly increased the thickness measurements and meatal mitotic index. Cerumol also increased the thickness measurements and the mitotic index; there was additionally low grade chronic inflammation in the dermis. Prophylene glycol increased the mitotic index and thickness measurements though no signs of inflammation were seen. Other compounds did not produce significant changes except that hydrocortisone reduced the mitotic index. Contact dermatitis to benzalkonium chloride may be important in persistent ear disease. Cerumenolytic agents ought not to be used for prolonged periods or as prophylaxis. PMID- 2970903 TI - The pathogenesis of chronic recurrent parotitis in infants: a study of 93 cases including an analysis of the vascular and glandular changes before and after parasympathectomy. AB - Ninety-three cases of infantile chronic recurrent parotitis are recorded. Fifty three cases were treated with parasympathectomy which was carried out at 2 levels. Good results were achieved in 79.1% of patients. The paper describes the use of the Doppler examination to study the effects of operation on the physiology of the parotid. PMID- 2970904 TI - Comparative cardiac effects of intravenous bolus of ipratropium bromide (itrop) and atropine sulfate in 22 patients. AB - At the present time, there is no satisfactory pharmacological treatment for arrhythmia or conduction disorders induced by or aggravated by vagal hypertonia. The limited duration of action of the atropine derivatives currently available justifies the development of new compounds with expected longer acting duration. The aim of this study was to compare the effects of a single blind intravenous injection of ipratropium bromide to those of atropine sulfate in 22 patients. These patients were studied with continuous Holter recordings for three days. During the second and the third nights (patient sleeping), boluses of atropine (0.03 mg/kg) and of ipratropium bromide (0.03 mg/kg), respectively, were added to a continuous saline intravenous infusion. Accurate ECG analysis allowed determination of maximal heart rate peak, timing of maximal heart rate, variations in sinus cycle length, atrioventricular conduction, and durations of drug action. A nonsuggestive questionnaire was presented to patients to detect possible occurrence of side effects. The mean maximal heart rate rose significantly (p less than 0.001) for atropine (+46.2%) and for ipratropium bromide (+57.4%). The effects obtained with ipratropium bromide on the heart rate lasted nearly twice as long as those obtained with atropine (respectively, 120 +/ 38.4 min and 70 +/- 30 min- for the pharmacological half-life). Common minor muscarinic side effects (dryness of the mouth) were noted with the two drugs. In conclusion, this comparative intraindividual study confirmed the prolonged vagolytic effects of intravenous ipratropium bromide, which may be valuable in the treatment of patients with vagally mediated automaticity and conduction disturbances. PMID- 2970905 TI - Antenatal and perinatal causes of handicap. PMID- 2970906 TI - Genetic causes of mental handicap and opportunities for prevention. AB - Prior to 1970 nearly one half of severe mental handicap (IQ less than 50) was unexplained and genetic causes were recognized in one third of all patients. In recent years the genetic contribution has risen to 50%, with the decline in importance of environmental causes and the growing recognition of X-linked forms of mental retardation and chromosomal microdeletion syndromes. Recognition of genetic conditions is vital for accurate assessment of recurrence risks and in order, in some instances, to provide specific prenatal diagnosis. With full utilization of existing prenatal screening procedures and a more detailed family history, severe mental handicap due to genetic conditions should be reduced by 30 40%. PMID- 2970907 TI - Porous polymers for biological fixation. AB - Porous polymers are recommended as coatings for femoral stems on the basis of their low modulus, which allows uniform distribution of stress to surrounding bone and the fabrication of more flexible stems by reducing the metallic cross section. Application of the porous polymer coating to the metallic substrate reduces metal ion release; the polymer coating does not affect the mechanical properties of the device. Porous polytetrafluoroethylene-carbon fiber composite, (PCFC), porous polyethylene, and porous polysulfone are undergoing investigation for this application. However, the low strength of the PCFC makes its use for this application questionable. Recently reported unfavorable clinical results (i.e., a high incidence of pain) have led to the discontinuation of one trial of porous polyethylene. Clinical investigation of porous polysulfone-coated titanium alloy devices are in progress. Animal studies have shown bone ingrowth into porous polyethylene and porous polysulfone implants in cortical and cancellous bone and in canine femoral stems coated with these materials. Pull-out testing of porous polysulfone implants revealed that the strength of the porous material was higher than the strength of surrounding bone. Less cortical bone loss was found around porous polysulfone-coated canine stems than was reported for fully coated porous metallic prostheses, presumably because of the lower stiffness. PMID- 2970908 TI - Down syndrome. Cervical spine abnormalities and problems. AB - A review of the radiographs of 34 individuals with Down syndrome (DS), between 5 and 21 years of age, demonstrated subluxation of atlantoaxial instability (C1-C2) greater than 5 mm in three of the 34 individuals (9 percent). This is in general agreement with previously reported ranges of 10 to 20 percent. A review of the cervical spine radiographs of adults with DS, between 26 and 42 years of age, showed no subluxation but significant degenerative changes of the cervical spine with spur formation, narrowing of foramina, narrowing of the disc inner space, and osteophyte formation. In both the DS child and the DS adult, the cervical spine may be an area of significant potential problems. In all cases any complaints of cervical pain or historical/clinical findings suggestive of neurologic involvement should initiate evaluation of the cervical spine. Baseline radiographic studies of the cervical spine are indicated in all DS children older than 5 years of age and should be considered in all DS adults, particularly those 30 years of age and older. PMID- 2970909 TI - Home nebulizers in childhood asthma. Parental perceptions and practices. AB - Seventy five percent of 91 parents of asthmatic children (aged 6 months to 15.4 years) with home nebulizers, responded to a questionnaire that sought to document parental experience with this form of therapy. The most common indications for acquiring a nebulizer were inability to use metered dose inhalers and poor response or intolerance to oral medications. When the children were classified into those who used the nebulizer daily (27/69) and those who used it less than daily (42/69), we found that the daily users were more likely to be in the severest category of asthma, by parental assessment, prior to the home nebulization program (p = 0.0035). Retrospective, uncontrolled comparison before and after the acquisition of a home nebulizer showed significant decreases both in hospital admissions (1.7 vs 0.7; p less than 0.001) and total in-patient days (4.1 vs 1.7; p less than 0.0001). Home nebulizers are well tolerated by parents and children alike and may be associated with decreased morbidity. PMID- 2970910 TI - Absorption and metabolism of nafarelin, a potent agonist of gonadotropin releasing hormone. AB - Nafarelin, a potent gonadotropin-releasing hormone (GnRH) agonist, was absorbed rapidly into systemic circulation (time to reach peak concentration, 20 to 40 minutes) after intranasal but not after sublingual or vaginal administration. Serum elimination half-life was about 2 hours. Nasal absorption of nafarelin was increased by increasing the concentration of the drug in the dose solution and incorporating sodium glycocholate into the nasal formulation. An optimal formulation providing maximum nasal absorption of nafarelin was one containing 1.75 mg nafarelin per ml and 2% sodium glycocholate. Bioavailability of this nasal formulation relative to a single subcutaneous dose averaged 21%. The metabolism and excretion of nafarelin were determined in three subjects after subcutaneous administration of [14C]-nafarelin. Radioactivity was excreted in approximately equal amounts in urine and stool. Six metabolites accounted for most of the radioactivity in urine. Four metabolites were short peptide fragments of nafarelin and the other metabolites were naphthylalanine and 2-naphthylacetic acid. PMID- 2970911 TI - Atrial natriuretic peptide and adaptation of sodium urinary excretion in patients with chronic renal failure. AB - 1. In order to examine the potential role of endogenous atrial natriuretic peptide (ANP) in modulating the increased sodium excretion per nephron in chronic renal failure, we studied healthy subjects with normal renal function (group I) and patients with moderate (group II) or severe chronic renal failure (group III) before, during and after administration of an intravenous sodium load. All subjects had been on a controlled diet containing 120 mmol of sodium per day for 5 days before the study. 2. Under basal conditions, plasma ANP and fractional excretion of sodium (FENa) were highest in group III. Both parameters increased in response to the sodium load in the three groups studied (P less than 0.001). Changes with time differed from group to group (P less than 0.05), the more marked response for both parameters being observed in group III. After adjustment with respect to plasma ANP (analysis of covariance), FENa was no longer modified in response to the sodium load, whereas adjustment of FENa with respect to mean blood pressure was without consequence on the significance of its change with time. This demonstrates that plasma ANP, but not mean blood pressure, represents the main factor producing variation in FENa during and after the sodium load. 3. These results suggest an important role for plasma ANP in promoting adaptation of short-term sodium excretion in response to an acute sodium load in patients with chronic renal failure who ingest a normal sodium intake. PMID- 2970913 TI - Osteopenia and diabetes. PMID- 2970912 TI - Phenotypes of peripheral blood T lymphocytes in rheumatoid arthritis and juvenile rheumatoid arthritis. Findings in patients with varying disease activity and clinical subgroups. AB - The percentage of T4 and T8 positive cells in peripheral blood of 78 patients with rheumatoid arthritis (RA) and 26 patients with juvenile rheumatoid arthritis (JRA) was determined using a rosette technique. The T4/T8 ratio +/- SEM (standard error of the mean) in the RA patients was increased, 2.02 +/- 0.09, as compared with normal blood donors, 1.71 +/- 0.06 (p less than 0.02). The subgroups of RA patients who had a significantly increased T4/T8 ratio were ANA positive patients (p less than 0.02) and patients on no medication (p less than 0.05). In the total group of JRA patients the T4/T8 ratio was 2.01 +/- 0.12 versus 1.75 +/- 0.08 in controls (p = 0.061). Polyarticular JRA patients had an increased T4/T8 ratio as compared with controls (p less than 0.05) while patients with the pauciarticular form had a normal ratio (p greater than 0.10). No correlation between the T4/T8 ratio and a disease outcome index, a laboratory activity index, ESR, WBC, platelet count, hemoglobin, serum albumin, age and disease duration was found (K less than +/- 0.20, p greater than 0.10). PMID- 2970915 TI - Basal rate subcutaneous insulin infusion: absorption kinetics and relation to local blood flow. AB - Twenty diabetic patients received a constant subcutaneous infusion of 125I labelled short-acting insulin (1.12 U/h) into the anterior abdominal wall. Measurement of the size of the insulin depot at the infusion site was performed by measuring the radioactivity and by using a calibration bolus to transform count rate to units of insulin in the depot. By the 133Xenon technique, local subcutaneous blood flow rate was measured on the contralateral side of the anterior abdominal wall. From the start of the infusion the SC depot built up slowly, but did not reach steady-state by 6 h. After 24 h of infusion, the size of the depot ranged between 2.7 and 10.9 U (5.2 +/- 0.5 U, mean +/- SE). The size of this steady-state depot was inversely correlated with SC blood flow. After stopping the infusion, insulin disappeared according to first order kinetics without any lag period. The insulin disappearance rate from the depot was positively correlated with the blood flow in a curvilinear fashion. The results suggest that the relationship between blood flow and the size of the insulin depot is best explained by a predominantly blood flow limited removal of insulin at low rates of blood flow, with increasing limitation by diffusion at higher rates. PMID- 2970914 TI - The effects of physical training in people with insulin-dependent diabetes. PMID- 2970916 TI - Insulin antibodies in diabetic children before treatment: a marker for islet B cell destruction? AB - Insulin antibodies were measured in the sera of 28 newly diagnosed diabetic children (age 8.0 +/- 4.0 (+/- SD) years) prior to insulin therapy and after 3, 6, 9, and 12 months. The levels at diagnosis and after 12 months were compared to endogenous insulin production at onset and after 12 to 14 months. Endogenous insulin production was evaluated through the measurement of 24-h urinary C peptide excretion, fasting plasma C-peptide levels and plasma C-peptide levels after glucagon stimulation. Insulin antibodies were detected in 29% of the patients (8 out of 28). In all but one patient antibodies binding porcine and human insulin were detected. No relationship was found between the presence of antibodies binding human or porcine insulin at diagnosis and age. After 1 year 27 out of 28 patients presented insulin antibodies. No relationship was found between the presence of insulin antibodies before therapy and 1 year after therapy. Insulin antibodies prior to diagnosis showed no relationship with the urinary C-peptide excretion at diagnosis (with antibodies 67 +/- 27%, without antibodies 76 +/- 11%). However, after 1 year significantly lower urinary C peptide excretions were found in patients with insulin antibodies prior to therapy (with antibodies, 17 +/- 7%, without antibodies, 31 +/- 5%, p less than 0.02). Peak plasma C-peptide levels after 1 year were possibly lower in patients with insulin antibodies before treatment (with antibodies 0.17 +/- 0.06 nmol/l, without antibodies 0.26 +/- 0.04 nmol/l, p less than 0.1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970917 TI - UK Prospective Diabetes Study. V. Characteristics of newly presenting type 2 diabetic patients: estimated insulin sensitivity and islet 4-cell function. Multi centre study. AB - In 713 newly diagnosed Caucasian diabetic patients aged 25-65 inclusive, insulin sensitivity and islet B-cell function were estimated from fasting plasma glucose and insulin concentrations by Homeostasis Model Assessment. Insulin sensitivity was reduced in obese subjects. It was also slightly lower in male than in female diabetic patients, in those who were sedentary and in those with high fasting plasma glucose concentrations. The estimated B-cell function was particularly impaired in patients with a high fasting plasma glucose and in those with normal rather than excess body weight. Whilst diabetes can present in normal weight patients with marked deficiency of B-cell function, presenting patients often have only a moderate impairment of B-cell function with markedly impaired insulin sensitivity secondary to obesity, physical inactivity, or being male. PMID- 2970918 TI - Health attitudes, self-monitoring of blood glucose, metabolic control and residual insulin secretion in type 1 diabetic patients. AB - The aim of the present cross-sectional study was to determine in patients who have had insulin-dependent diabetes mellitus for a long time, the relationship between residual insulin secretion and self-monitoring of blood glucose, psychological factors, and blood glucose control (HbA1c). In 97 patients with a disease duration of 9-16 years a sample from a 24-h urine collection was analysed for C-peptide excretion. In 35 of these patients an excretion greater than or equal to 0.2 nmol/24 h was detected. They were matched for age at onset and disease duration with patients without any detectable C-peptide excretion. Twenty nine matched pairs were obtained and 22 of them have been further investigated. HbA1c was lower in the C-peptide excretors than in the non-excretors (6.9 +/- 0.3 vs 7.9 +/- 0.3%) (p less than 0.025). The patients were interviewed by means of a questionnaire dealing with their 'health beliefs'. It was shown that in the group with residual insulin secretion there was a significantly lower perceived risk of developing kidney disease (p less than 0.025) than in the group without residual insulin secretion, but in all other items measured there was no difference between the two groups. Furthermore, in the group with residual insulin secretion we found a correlation (p less than 0.01) of low HbA1c with high frequency of self-monitoring of blood glucose and high perceived vulnerability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970919 TI - Reduced venous compliance and increased transcapillary escape of protein in insulin-dependent diabetic patients. AB - Lower limb venous compliance and transcapillary escape rate of transferrin were measured in eight normotensive, insulin-dependent male diabetic patients and eight control subjects using a dual isotope technique. Technetium-99m labelled autologous erythrocytes were used to measure venous compliance and to correct for local changes in blood volume, whilst Indium-113m labelled transferrin was used to measure transcapillary escape of protein. The diabetic patients were found to have reduced venous compliance 1.5 (0.7 to 3.4) x 10(-2) mmHg-1 compared with controls 3.2 (2.4 to 4.1) x 10(-2) mmHg-1 (p less than 0.01). The diabetic patients were also found to have greater transcapillary escape of transferrin 2.7 (-1.5 to -5.3) x 10(-3), compared with control subjects -5.2 (-4.1 to -8.1) x 10(-3) (p less than 0.02) in response to increasing hydrostatic pressure. These results show reduced venous compliance in patients with a mean duration of diabetes of 15 years and with only at most, early complications of diabetes, and confirm previous observations showing increased transcapillary escape of protein. PMID- 2970920 TI - Use of memory meters to measure reliability of self blood glucose monitoring. AB - Glucose reflectance meters equipped with memories were used in 21 insulin dependent diabetic patients to assess reliability of self blood glucose monitoring records. Results reported in logbooks showed widespread discrepancies when compared with corresponding meter-held records. Discrepancy rates of at least one in five were observed in 11 patients. Unreliable reporting was usually recognized by clinical staff, and associated with high glycosylated haemoglobin levels and poor technique in measuring blood glucose at home. PMID- 2970921 TI - Urinary ethanol and diabetes mellitus. AB - Ethanol concentrations were assayed in urine samples obtained from 10 newly presenting diabetic patients (aged 50-69 years) with glycosuria and symptoms suggesting genito-urinary candidiasis. No alcohol was detected initially with diabetic urine samples, but when stored at room temperature 8 out of 10 samples showed increasing levels of ethanol from day 3 to day 20. Peak levels were more than 10,000 mg/l in three samples and more than 1500 mg/l in six samples. Studies were also performed using normal urine and urine to which was added glucose and/or Candida. Samples were kept at either room temperature or 4 degrees C for up to 21 days and ethanol assays performed at regular intervals. Studies with non diabetic urine showed a maximum ethanol level of 500 mg/l after 11 days only in samples containing both glucose and Candida and kept at room temperature. PMID- 2970922 TI - Scleroedema diabeticorum: a minor but often unrecognized complication of diabetes mellitus. AB - The association of specific dermatological complications with diabetes mellitus is well recognized. Of 100 hospital-based patients with diabetes mellitus (age 48 years +/- 2SE), 14% had scleroedema diabeticorum. The affected subjects had a higher prevalence of retinopathy (p less than 0.001) and albuminuria (p less than 0.025). The duration of scleroedema correlated with the duration of diabetes (p less than 0.005). These findings highlight the relatively common occurrence of this skin condition which often goes unrecognized in people with diabetes. PMID- 2970923 TI - Platelet function in uncomplicated insulin-dependent diabetic patients at rest and following exercise. AB - There are conflicting reports of platelet function abnormalities in diabetic patients without vascular complications. We have studied in vitro platelet aggregation, using platelet rich plasma and whole blood techniques, in 18 patients with uncomplicated insulin-dependent diabetes and a matched group of 24 non-diabetic subjects. In addition we measured plasma beta-thromboglobulin levels in these groups, as an index of in vivo platelet activation, and compared the indices of in vitro and in vivo platelet function before and after maximal bicycle exercise. Before exercise plasma beta-thromboglobulin levels and platelet sensitivities to ADP, collagen or adrenaline, as assessed by both methods of platelet aggregation, were the same in diabetic and control subjects. Both groups showed similar increases in beta-thromboglobulin levels and in platelet sensitivity to all agonists in whole blood following exercise. Using platelet rich plasma there were no changes in platelet sensitivity in either group after exercise. In non-diabetic subjects, increases in noradrenaline levels after exercise correlated with increases in platelet sensitivity to adrenaline in whole blood. This was not observed in the diabetic group. Abnormalities of platelet function, using the techniques described here, are not present in diabetic patients who do not have clinical evidence of vascular disease. PMID- 2970924 TI - An evaluation of multiple choice questionnaires for the assessment of knowledge in diabetic children and their families. AB - The comparability of five multiple choice questionnaires was tested in 69 diabetic children and their parents. Analysis of variance showed that 68% of the variation in the scores was due to differences between individuals, with only 8% due to differences between questionnaires. A second study then compared results from diabetic children, their mothers and fathers, and some nurses and doctors, who each completed one questionnaire. Analysis of variance of this study showed that the mothers, fathers, and nurses scored about the same, but more than the children (p less than 0.05), whilst the scores of the doctors were not significantly different from any of the other groups. PMID- 2970925 TI - A proficiency score for assessing the reliability of self blood glucose monitoring. AB - The reliability of self blood glucose analysis was studied in 83 insulin-treated diabetic patients. Reliability was assessed by laboratory analysis of capillary blood samples collected simultaneously on filter paper. Patient and laboratory results were compared using a scoring system which produces an easily understood Proficiency Score. The score was designed to highlight unreliable performance in measuring glucose concentrations in the euglycaemic as well as hypo- and hyperglycaemic ranges. Only 17% of patients achieved acceptable Proficiency Scores by maintaining adequate performance on five occasions. PMID- 2970926 TI - New referrals to a diabetic clinic. AB - New referrals to a diabetic clinic were studied over 6 months. One hundred and twelve patients were referred from their General Practitioner (GP) and 49 from other hospital departments. Thirty percent of GP referrals and 10% of hospital referrals were made on the basis of glycosuria alone. From all the patients referred six were found to be non-diabetic and a further 19 had been prescribed drugs inappropriately (either the wrong drug or the right drug in excessive dose), some with potentially dangerous consequences. Almost half of the patients were given no dietary advice at all. There is, still much ignorance concerning diagnosis and initial management of diabetes in both general and hospital medical practice. PMID- 2970927 TI - Haemoglobin K-Woolwich as cause of falsely elevated HbA1. AB - Falsely elevated levels for haemoglobin A1 can be found with the haemoglobinopathies. A case is reported where falsely high levels for haemoglobin A1 were caused by the asymptomatic presence of haemoglobin K-Woolwich. PMID- 2970928 TI - Variability of albumin excretion in children. PMID- 2970929 TI - Responses to chickling vetch and bread meals in normal and type 2 non-insulin dependent diabetic subjects. PMID- 2970930 TI - Social iatrogenesis of the Australian disease 'RSI'. PMID- 2970931 TI - Primary irritant effect of 3,4,5-trichloropyridazine. AB - A primary irritant dermatitis to 3,4,5-trichloropyridazine was observed on the hands of a chemist after accidental exposure. Healing was rapid in comparison to a report on 3,4,6-trichloropyridazine toxicity. Prompt treatment with a potent topical corticosteroid, wet soaks, and an oral antihistamine brought resolution within 14 days. PMID- 2970932 TI - Contact dermatitis in funeral service workers. AB - 84 funeral service workers and 38 control workers were evaluated for the presence of skin disease by history, clinical examination and patch tests with formaldehyde and glutaraldehyde. No relationship between either personal or family history of cutaneous or respiratory manifestations of atopy and clinical parameters of cutaneous disease or patch test results was found. Cutaneous disease was reported in apprentices, active embalmers and inactive embalmers in decreasing order of frequency. Positive patch test reactions to formaldehyde and glutaraldehyde were found in 4% and 7% of the exposed workers, but in none of the controls. Although exposure to glutaraldehyde was less frequent, the prevalence of positive patch test reactions did not differ. This may suggest that glutaraldehyde poses a greater practical risk of cutaneous sensitization in this trade than formaldehyde. PMID- 2970933 TI - Frequency of sensitisation to the preservative Kathon CG. AB - The incidences of sensitisation to Kathon CG are reported for patients routinely tested from 1984 to 1986 in Bari, London, Louvain, Nijmegen, Oulu and San Francisco. For these 3 years, the overall frequency of sensitisation varied for women from 3.3% in Oulu to 0.6% in London and San Francisco and for men from 1.4% in Oulu to 0% in San Francisco. Women were predominantly sensitised, probably by cosmetics, toiletries and domestic cleaners. Occupational sensitisation was much less frequent. PMID- 2970934 TI - Irritant contact dermatitis from diallylglycol carbonate monomer and its prevention. AB - We report an outbreak of irritant contact dermatitis from diallylglycol carbonate monomer among factory employees making optical plastic lenses. Symptoms lessened after simple preventive methods were introduced. Further upgrading of production methods lessened symptoms still further and also restored the company's economic viability. PMID- 2970936 TI - Occupational allergic contact dermatitis from Rengas wood. PMID- 2970935 TI - Allergic dermatoses and respiratory diseases from reactive dyes. AB - 5 cases of occupational eczema, urticaria and respiratory disease from reactive dyes, occurring during 1977-1987, are reported. The patients, 4 men and 1 woman, were 24-52 years old when examined. They had been working in dye houses or textile plants, and had been exposed to reactive dyes for 8 months to 4 years before symptoms developed. Only 1 of the patients has been able to continue in the same occupation. On patch testing, the 4 patients with eczema reacted positively to 9 commercial dye powders. 2 patients reacted to the same dye, Remazol Schwarz B. On scratch and/or prick testing, the 2 patients who also had respiratory symptoms and/or urticaria reacted positively to the same dyes as on patch testing. The 5th patient, who had urticaria and respiratory symptoms, reacted positively to a dye, Remazol Gold Gelb RNL, but the patch test with that dye was negative. None of the patients was patch-test-positive to para phenylenediamine (PPD) or to textile dye allergens in a series of organic dyes. Thus, the series of organic dyes has little value in the screening of allergy to reactive dyes. A 1% pet. dilution of commercial dye powder for patch testing and the same concentration in distilled water for prick testing seem to be suitable for the screening of allergy to reactive dyes. PMID- 2970937 TI - Contact allergy to benzyl alcohol in a cutting oil reodorant. PMID- 2970938 TI - Chloracetamide as an allergen in the paint industry. PMID- 2970939 TI - Sensitization to DD soil fumigant during manufacture. PMID- 2970940 TI - Metal content of Indian coins. PMID- 2970941 TI - Facial dermatitis from a neoprene rubber mask. PMID- 2970942 TI - Contact granuloma annulare. PMID- 2970943 TI - Photoallergic contact dermatitis from fentichlor. PMID- 2970944 TI - Allergic contact dermatitis from dodecyldiaminoethyl glycine. AB - We were previously ignorant of DDEG and its widespread uses. We have been unable to find references to any previous cases and suggest a 1% aq. concentration of it as suitable for patch testing. PMID- 2970945 TI - The immunobiology of the T cell response to Mls locus disparate stimulator cells. PMID- 2970946 TI - Short-term treatment of dandruff with a combination of propylene glycol solution and shampoo. AB - Many studies now indicate an association between the lipophilic dimorphic yeast Pityrosporon ovale and dandruff. Propylene glycol has been proved effective in the treatment of pityriasis versicolor, Pityrosporon folliculitis, and seborrheic dermatitis. The patients in this study found treatment with a combination of propylene glycol, ethanol, and water, used with a shampoo, to be effective, easy to use, and cosmetically attractive. No side effects were observed by the patients. PMID- 2970947 TI - Highlights of the AAD post-graduate course "Recent Developments in Contact Dermatitis and Occupational Dermatology" sponsored by the AAD with the North American Contact Dermatitis Group San Diego, May 21-28, 1988. Part I. PMID- 2970948 TI - Toxicodendron dermatitis. PMID- 2970949 TI - Pharmacokinetics of teicoplanin. AB - We investigated the pharmacokinetic properties of teicoplanin, after 200 mg i.v. and i.m. administration in 10 healthy male subjects by assuming a three compartment open model with elimination from the central compartment. The mean peak plasma level was 7.16 micrograms/ml reached after 2.26 h. The half-life, the plasma and renal clearances, evaluated from i.v. data were 44.49 h, 15.31 and 9.08 ml/min, respectively. The same parameters after i.m. administration were 45.62 h, 15.31 and 9.46 ml/min. The estimates of creatinine clearance (Clcr greater than 80 ml/min), renal clearance and the low free fraction (fB approximately equal to 0.1) suggested a tubular reabsorption, FR, of the drug. The distribution volume at steady state after i.v. and i.m. administration (Vss = 41.29 and 44.76 litres) were nearly total body water. Bioavailability of the drug (F = 92.4%) showed an almost completely absorption of teicoplanin after i.m. administration. Urinary recovery was 49.6 and 47.9% of the dose after i.v. and i.m. administration, respectively. PMID- 2970950 TI - High teicoplanin uptake by human neutrophils. AB - The cellular/extracellular (C/E) concentration ratio of teicoplanin in polymorphonuclear leukocytes (PMNs) increased rapidly with time (C/E 60 +/- 13 at 20 min). The C/E ratio was time- and concentration-dependent. At 20 min and an initial concentration of 75 +/- 16 micrograms/ml the cellular drug concentration was 4,700 +/- 1,300 micrograms/ml. The mechanism of drug uptake was by an active process and transported (cellular) drug retained its antimicrobial activity. Washing removed 42% of cellular drug. Teicoplanin inhibited PMN chemotaxis at very high concentrations and PMN microbicidal activity at lower concentrations. PMID- 2970951 TI - Cefodizime administration in healthy subjects: studies of natural killer cells, urinary hormones and electrolytes. AB - Two studies are here discussed: the first one on changes of the natural killer cell activity of PBM cells exposed in vitro to CDZ and the second one on urinary electrolytes, cortisol, aldosterone and DHEA-S circadian rhythms, evaluated in healthy subjects who received a single dose (2 g i.v.) of cefodizime (CDZ). The effect of CDZ in health includes a reduction of the circadian amplitude of natural killer cell activity. With the methods used, no difference was found between placebo and CDZ, as far as circadian rhythms of urinary electrolytes, cortisol, aldosterone and DHEA-S are concerned. PMID- 2970952 TI - [Trichothecenes in staple food from a high-incidence area of carcinoma of the esophagus and gastric cardia and their carcinogenic potential]. AB - This is the first report to demonstrate the presence of a group of mycotoxins in corn-staple food of Linxian residents, including Nivalenoal (NIV), Deoxynivalenol (DON), 3-acetyl-deoxynivalenol (3-ADON), 15-acetyl-deoxynivalenol (15-ADON), T-2 toxin (these 5 toxins belong to trichothecenes) and zearalenone (ZEN). These 6 toxins were produced by Fusarium species. Using TLC, HPLC, GC, monoclonal antibody affinity column chromatography combined with RIA, respectively, 107 corn samples from Linxian and 2 rice samples from USA were analyzed. It was demonstrated that the average level of NIV was 757 +/- 707(54-2,760)ng/g, DON was 5,376 +/- 4,460 (360-12,670) ng/g with 100% positivity in corn samples consumed as staple food by esophageal cancer patients and their families. Corn samples collected from 5 villages in Linxian at different seasons in 1984-1986 all showed high levels of NIV and DON with 100% positivity which suggested that NIV and DON were consistently and widely existed in Linxian corn samples. Levels of 3-ADON and 15-ADON in Linxian corn were 113 +/- 57 and 495 +/- 538 ng/g. This paper also first demonstrates that the extracts of corn from esophageal cancer patients' families and their NIV and DON fractions purified by HPLC could induce significant chromosome aberrations in V79 cells, indicating that they had carcinogenic potential. Pure toxins of NIV, DON, T-2 and 3-ADON could induce chromosome aberrations in V79 cells starting from 1 ng/ml-a very low concentration. When concentrations increased to certain range they all showed toxic effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970953 TI - [Abdominal wall desmoid tumor--analysis of 42 patients]. AB - Forty two patients with abdominal wall desmoid tumor, including one Gardner's syndrome, are reported. All patients were female except one. The tumor occurred in various sites in the abdominal wall, 66% in the lower abdominal wall. The fascia, sheath and muscle layer were chiefly involved and a very large tumor could invade peritoneum and viscera. This tumor shows aggressive growth and is prone to recurrence. Surgery is the treatment of choice. Local recurrence rate was 5.5%. Abdominal wall desmoid tumor associated colonic polyposis is named Gardner's syndrome. The authors emphasize that a local extended resection should be performed with a safety margin at least 2-3 cm beyond the tumor. Peritoneum, if involved, should be resected together with the primary focus. PMID- 2970954 TI - [Synthesis and comparative biological activity of atrial natriuretic peptide analogs]. PMID- 2970955 TI - Comparison of cicletanine with other antihypertensive drugs in SHR-SP models. AB - The effects of cicletanine were compared with those of four other antihypertensive drugs (prazosin, a highly selective alpha 1 antagonist; captopril, an angiotensin-converting enzyme inhibitor; indapamide, an antihypertensive diuretic; and hydrochlorothiazide, a purely diuretic agent) on young stroke-prone SHR rats with high-salt diet. All the drugs except hydrochlorothiazide prevented the onset of hypertension. The minimal effective dose on blood pressure was 1 mg/kg for both cicletanine and captopril, and 3 mg/kg for indapamide. The action on cardiac hypertrophy and diuresis occurs at a dose of cicletanine 10 to 30 times higher than that required to produce the antihypertensive effect. Renal hypertrophy was also decreased significantly by cicletanine at a dose of 100 mg/kg. PMID- 2970956 TI - Modrasone for the skin: worth prescribing? PMID- 2970957 TI - [Fetal and neonatal blood flow]. PMID- 2970958 TI - Insulin-like growth factor-binding proteins (IGF-BPs) produced by human skin fibroblasts: immunological relationship to other human IGF-BPs. AB - We have characterized the insulin-like growth factor-binding protein (IGF-BP) produced by neonatal human skin fibroblasts in monolayer culture using antibodies specific for the acid-stable subunit of the 150K GH-dependent IGF-BP complex, BP 53, and the amniotic fluid IGF-BP, BP-28. Fibroblasts produced 65.3 +/- 10.4 ng/ml.72 h (SE; n = 6) immunoreactive BP-53 in serum-free medium; this was stimulated by increasing fetal bovine serum in the medium up to 385.3 +/- 49.0 ng/ml.72 h at 10% serum. Epidermal growth factor (EGF) also caused dose-dependent stimulation of BP-53 production, with a maximal effect (3-fold increase) at 30 ng/ml EGF. No immunoreactive BP-28 production was detectable in the presence or absence of serum or EGF. Neutral gel chromatography of serum-free medium revealed a peak of immunoreactive BP-53 at about 50K, with a smaller species at 20-30 K. Serum- and EGF-stimulated cells produced higher levels of about 50K BP-53, and an additional peak of immunoreactivity at 150K was present in serum-stimulated, but not EGF-stimulated, samples. Comparison of IGF-I and IGF-II binding by fibroblast BP-53 revealed slightly higher IGF-II than IGF-I binding, and association constants of 3-4 x 10(10) liter/mol for both IGFs, similar to BP-53 from human plasma. Affinity labeling of acid-stripped medium followed by nonreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed specifically cross linked IGF-binding species of 60K (identical to labeled plasma BP-53), 42K, and 37K. Only the 60K and 42K complexes were precipitable by antiserum to plasma BP 53, and none was precipitable by anti-BP-28 serum, suggesting that the 37K band might represent a third class of IGF-BP. We conclude that neonatal skin fibroblasts produce no BP-28, but do produce two IGF-BPs immunologically homologous to human plasma BP-53, one of which shows size and IGF-binding characteristics identical to the plasma protein. PMID- 2970959 TI - Acetylcholine and norepinephrine stimulate the release of corticotropin-releasing factor-41 from the rat hypothalamus in vitro. AB - The effects of the two putative neurotransmitters acetylcholine and norepinephrine on immunoreactive CRF-41 release from incubated rat hypothalami were studied. Acetylcholine at concentrations of 10(-11) to 10(-7) M stimulated CRF-41 release. This effect was blocked in a dose-dependent manner by the muscarinic antagonist atropine (10(-9) to (-7) M). The nicotinic antagonist hexamethonium was ineffective at a dose of 10(-7) M, but produced slight inhibition of this response at 10(-5) M. Norepinephrine at concentrations of 10( 10) to 10(-6) M also produced a dose-dependent stimulation of CRF-41 release. The beta-adrenoceptor antagonists propranolol (10(-5) M) and timolol (10(-6) M) blocked norepinephrine-induced CRF-41 release. The alpha 1-adrenoceptor antagonists thymoxamine (10(-5) M), prazosin (10(-5) M), and corynanthine (10(-4) M), and the alpha 2-antagonist idazoxan (10(-5) M), were ineffective. Potassium depolarization (56 mM) caused stimulation of CRF-41 release which was dependent on the presence of calcium in the incubation medium. Authenticity of immunoreactive CRF-41 released was demonstrated by chromatographic criteria using gel filtration and reversed phase HPLC. These results provide evidence for a stimulatory role of acetylcholine and norepinephrine on CRF-41 release, and consequently on hypothalamo-pituitary-adrenal axis in the rat, through actions at a hypothalamic level. The stimulatory effect of acetylcholine is mediated principally through muscarinic receptors and that of norepinephrine through beta adrenoceptors. PMID- 2970960 TI - Cellular processing of pro-atrial natriuretic factor (pro-ANF): studies using an antiserum that selectively binds ANF-(99-126) after its cleavage from pro-ANF. AB - Atrial natriuretic factor (ANF) is stored in atrial myocytes as a 15-17K prohormone, but circulates in plasma as a 3K, carboxy (C)-terminal fragment of the prohormone. The tissue location at which the cleavage of pro-ANF to its hormonal form occurs is unknown. In the present study, an immunological approach was taken to address this question. A polyclonal antiserum was generated which recognizes the hormonal form of ANF [ANF-(99-126)] only after its cleavage from the prohormone. This was accomplished by immunizing rabbits with a synthetic peptide corresponding to the seven amino (N)-terminal residues of ANF-(99-126) coupled to carrier protein via a C-terminal cysteine. This antiserum, anti-ANF (99-105), demonstrated high affinity for ANF-(99-126) (IC50 = 170 pM), but displayed 100-fold less affinity for recombinant pro-ANF [ANF-(2-126)]. The N terminal specificity of anti-ANF-(99-105) was evident by its failure to bind ANF (103-126) at concentrations up to 100 nM. The specificity of anti-ANF-(99-105) for the hormonal form of ANF was examined by using thrombin to cleave pro-ANF and testing for the generation of anti-ANF-(99-105) immunoreactivity. Cleavage of atrial pro-ANF or 35S biosynthetically-labeled pro-ANF resulted in the production of immunoreactive material from the prohormone, whereas pro-ANF itself demonstrated no cross-reactivity with anti-ANF-(99-105). Anti-ANF-(99-105) could also recognize ANF released from the isolated perfused rat heart. When anti-ANF (99-105) was used in immunohistochemical studies of rat atrial myocardium, no staining was observed in unfixed frozen sections. This suggests that proteolytic processing of pro-ANF is not an intracardiocytic event. PMID- 2970961 TI - Receptors for insulin-like growth factors I and II: autoradiographic localization in rat brain and comparison to receptors for insulin. AB - Receptors for insulin-like growth factor I (IGF-I) in rat brain were visualized using autoradiography with [125I]IGF-I. The binding of the labeled peptide was competed for fully by high concentrations of unlabeled IGF-I. At intermediate concentrations of unlabeled peptide the binding of [125I]IGF-I was competed for by unlabeled IGF-I more effectively than by IGF-II or insulin, which is typical of receptors for IGF-I. Essentially every brain section shows specific binding of IGF-I, and the pattern of binding of IGF-I to its receptors correlated well with the cytoarchitectonic structures. In parallel studies we showed that [125I]IGF-II was bound to tissue sections of rat brain and that the binding was competed for by an excess of unlabeled IGF-II. However, intermediate concentrations of unlabeled peptides gave inconclusive results. To confirm that the binding of [125I]IGF-II was to IGF-II receptors, we showed that antibodies specific for the IGF-II receptor inhibited the binding of labeled IGF-II. Furthermore, the binding of the antibody to regions of the brain section, visualized by the application of [125I]protein-A, gave patterns indistinguishable from those obtained with [125I]IGF-II alone. Again, the binding was very widely distributed throughout the central nervous system, and the patterns of distribution corresponded well to the underlying neural structures. Densitometric analysis of the receptors enabled us to compare the distribution of IGF-I receptors with that of IGF-II receptors as well as retrospectively with that of insulin receptors. PMID- 2970962 TI - Endocytosis of receptor-bound insulin-like growth factor II is enhanced by mannose-6-phosphate in IM9 cells. AB - The insulin-like growth factor II (IGF-II), and glycoproteins containing mannose 6-phosphate (M6P), bind to two different sites of the same receptor molecule (Morgan et al, Nature 329:301, 1987). To study the interactions between the two ligands on their common receptor in intact cells, we examined the effect of free M6P on IGF-II binding and endocytosis in the IM9 human lymphoblastoid cell line. M6P, up to a 3 mM concentration, had no effect on the binding of IGF-II to the cell surface receptor of intact IM9 cells at 4 degrees C. By contrast, when IM9 cells were incubated with 125I-IGF-II at 37 degrees C, 1mM M6P increased cell associated radioactivity by twofold. The increase was resistant to acid wash at 4 degrees C, and therefore assumed to represent endocytosed IGF-II. Acid-washable radioactivity was no different, confirming that, in intact cells, M6P does not affect IGF-II surface binding. In addition, preincubation of cells with M6P at 37 degrees C for up to 3 hours did not change the abundance of receptor on the cell surface, as measured by a subsequent 4 degrees C binding assay. We conclude that M6P causes a shift of IGF-II-occupied receptors form the cell surface to intracellular locations without affecting surface binding of this ligand in IM9 cells. The effect could be produced by the binding of M6P itself, or by the displacement of endogenous phosphomannosylated ligands. PMID- 2970964 TI - Calcium-dependent inactivation of the Ca2+-ATPase from sarcoplasmic reticulum by chemically reactive adenosine triphosphate. AB - 1. ATP gamma P-imidazolidate, synthesized from ATP and carbonyldiimidazole, inhibits the Ca2+-ATPase of sarcoplasmic reticulum in a biphasic manner. A fast first phase is concentration-dependent while a slower second phase is independent of the inhibitor concentration. 2. The inhibition is calcium- and magnesium dependent. No inhibition occurs in the absence of either cation. 3. Inhibition of the Ca2+-ATPase can be prevented by the protection with ATP. 4. The loss of ATPase activity is pH-dependent. Maximal inhibition coincides with maximal ATPase activity. It indicates a participation of the reacting amino acid side chain in the catalytic cycle. 5. The incorporation of radioactive inhibitor is reversed in a time-dependent fashion while the Ca2+-ATPase remains inhibited. 6. We conclude that ATP gamma P-imidazolidate initially reacts covalently with an amino acid side chain, probably Asp-351, but is subsequently expelled by a reaction with a second amino acid. 7. This two-step reaction induces an intramolecular cross-link which can be shown by the creation of a new protein band on SDS-PAGE which originates in the Ca2+-ATPase. PMID- 2970966 TI - Skin microvascular circulation in the sympathetic dystrophies evaluated by videophotometric capillaroscopy and laser Doppler fluxmetry. AB - Finger-skin microcirculation and its reactions to sympathetic stimuli were investigated in 12 patients with sympathetic dystrophies, secondary to trauma or other diseases. Nailfold-skin capillary blood cell velocity (CBV) was measured by videophotometric capillaroscopy. Laser Doppler fluxmetry was used to provide an index of skin circulation in vessels in addition to the superficial capillaries. Both CBV and laser Doppler flux (LDF) values were significantly lower in the patients, compared with the healthy controls (P less than 0.05), despite the fact that skin temperature was the same in both groups. During cooling of the contralateral hand, CBV and LDF decreased markedly (22-60%) in the control group but not in the patients (0-13%). The decrease in skin perfusion normally seen upon lowering of the hand was also impaired in the patient group (7%) compared with controls (42%) (P less than 0.05). These impaired vasomotor reflex responses are consistent with sympathetic dysfunction and may well explain some of the typical features of the syndrome, e.g. limb oedema. PMID- 2970965 TI - Glycogenin is the priming glucosyltransferase required for the initiation of glycogen biogenesis in rabbit skeletal muscle. AB - Purified preparations of glycogen synthase are a complex of two proteins, the catalytic subunit of glycogen synthase and glycogenin, present in a 1:1 molar ratio [J. Pitcher, C. Smythe, D. G. Campbell & P. Cohen (1987) Eur. J. Biochem. 169, 497-502]. This complex has now been found to contain a further glucosyltransferase activity that catalyses the transfer of glucose residues from UDP-Glc to glucosylated-glycogenin. The glucosyltransferase, which is of critical importance in forming the primer required for de novo glycogen biosynthesis, is distinct from glycogen synthase in several ways. It has an absolute requirement for divalent cations, a 1000-fold lower Km for UDP-Glc and its activity is unaffected by incubation with UDP-pyridoxal or exposure to 2 M LiBr, which inactivate glycogen synthase by 95% and 100%, respectively. The priming glucosyltransferase and glycogen synthase activities coelute on Superose 6, and the rate of glycosylation of glycogenin is independent of enzyme concentration, suggesting that the reaction is catalysed intramolecularly by a subunit of the glycogen synthase complex. This component has been identified as glycogenin, following dissociation of the subunits in 2 M LiBr and their separation on Superose 12. The glycosylation of isolated glycogenin reaches a plateau when five additional glucose residues have been added to the protein, and digestion with alpha-amylase indicates that all the glycogenin molecules contain at least one glucosyl residue prior to autoglucosylation. The priming glucosyltransferase activity of glycogenin is unaffected by either glucose 6-phosphate or by phosphorylation of the catalytic subunit of glycogen synthase. The mechanism of primer formation is discussed in the light of the finding that glycogenin is an enzyme that catalyses its own autoglucosylation. PMID- 2970963 TI - Molecular characterization and chromosomal mapping of melanoma growth stimulatory activity, a growth factor structurally related to beta-thromboglobulin. AB - Melanoma growth stimulatory activity (MGSA) is a mitogenic polypeptide secreted by Hs294T human melanoma cells. Comparison of the N-terminal sequences of the 13 and 16 kd MGSA species with the cDNA sequence revealed that the mature form of human MGSA is maximally 73 amino acids long. Expression of the cDNA in mammalian cells results in the secretion of this peptide with mitogenic activity. MGSA is structurally related to the platelet-derived beta-thromboglobulin and to several other polypeptides. These factors may constitute a family of growth factors. MGSA mRNA was detected in a variety of cell types. The level of MGSA mRNA in melanoma cells is strongly elevated by treatment with MGSA. MGSA is the gene product of a recently detected gene gro. The gene was mapped to chromosome 4 (region q13--- q21). This same region also contains genes for two of the structurally related factors, for c-kit, a receptor for an as yet unidentified ligand, and for 'piebald trait', an inherited skin pigmentation disorder. PMID- 2970967 TI - Modification of tumor cells by a low dose of Newcastle disease virus. Augmentation of the tumor-specific T cell response in the absence of an anti viral response. AB - The present study elucidates the mechanism whereby viral xenogenization of highly metastatic ESb lymphoid tumor cells increases tumor immunogenicity and syngeneic tumor-specific T cell responses in comparison to nonmodified tumor cells. It was found that the frequency of cytotoxic T lymphocytes specific for the Esb tumor associated transplantation antigen (TATA) and the cytotoxic anti-tumor activity in bulk cultures of immune spleen cells were significantly increased (by factor 3 and 25, respectively) when using virus-modified tumor cells. An amplified response was observed both in vivo and in vitro which might explain the demonstrated effectiveness of this approach for postoperative immunotherapy of ESb metastases. For the stimulation of tumor-specific cytolytic T lymphocytes (CTL) the ESb tumor cells which are highly metastatic were infected with an avirulent strain of the paramyxovirus Newcastle Disease Virus (NDV). Infection of ESb cells with low amounts of NDV was sufficient to lead to an increase in cytolytic activity of tumor-specific CTL after sensitization in vivo and restimulation in vitro. In a sensitive limiting dilution mixed leukocyte-tumor cell microculture system the direct effect of viral modification on the frequency and specificity of CTL was investigated. The number of ESb-specific CTL per spleen could be raised from about 3300 (without modification) to 9100 by both in vivo and in vitro application of ESb-NDV. One application of ESb-NDV (in vivo or in vitro) increased the number of CTL to 4900 and 4600, respectively. In split type experiments it could be shown at the clonal level that viral modification did not alter the specificity of ESb-specific CTL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970968 TI - Two subsets of human CD4+ T helper cells differing in kinetics and capacities to produce interleukin 2 and interferon-gamma can be defined by the Leu-18 and UCHL1 monoclonal antibodies. AB - Human CD4+ T helper cells were separated into CD4+45R+ and CD4+45R- cells. When stimulated with the polyclonal activator staphylococcal enterotoxin A in the presence of autologous monocytes, these two subsets exhibited a striking difference in production of interleukin 2 (IL2) and interferon-gamma (IFN-gamma). While the CD4+45R- subset produced maximal amounts of IL2 within 24 h and IFN gamma within 72 h, the CD4+45R+ subset produced no IL2 within 24 h and merely marginal amounts of IFN-gamma as assayed after 24 to 96 h. This discrepancy between the subsets was found when the cells were stimulated by other accessory cell-dependent activators and by the accessory-independent combination of the calcium ionophore A23187 and the phorbol ester 12-O-tetradecanoylphorbol 13 acetate as well. The inability of the CD4+45R+ cells to produce IL2 during the first day of culture was not due to any modulation of either the CD4 or the CD45R antigens, as purified CD4+45R+ cells obtained by negative panning selection with the reciprocal UCHL1 monoclonal antibody responded in a similar manner as the positively selected sorted CD4+45R+ cells. Analysis of the kinetics of IL2 production by the two T helper cell subsets clearly demonstrated that the IL2 recorded after 1 day of culture was entirely produced by the CD4+45R- cells, whereas the CD4+45R+ cells produced IL2 during and after the second day of culture. This discrepancy in kinetics was not due to an increased absorption of IL2 by the CD4+45R+ cells during the first day of culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970969 TI - The role of suppression in immunoregulation: in vivo analysis using a monoclonal antibody to T suppressor factors. AB - We have used a monoclonal antibody (mAb)-specific for murine T suppressor (Ts) cells (mAb 14-12) to study the role of T cells in tolerance and immunoregulation. We demonstrate that mAb 14-12 can block in vivo Ts cell activity in a variety of experimental systems. It prevents the induction of Ts cells induced by i.v. injection of the water-soluble hapten 2,4,6-trinitrobenzene sulfonic acid, and the protein antigen bovine serum albumin. When 14-12 mAb is given prior to the i.v. injection of trinitrophenyl-conjugated spleen cells (TNP-SC) it blocks the induction of Ts cells and sufficiently overcomes suppression so that TNP-SC is able to induce immunity. mAb 14-12 can convert nonresponder mice into responders for the Ir gene-controlled response to the random terpolymer L-glutamic acid60-L alanine30-L-tyrosine 10 (GAT), and can substitute for cyclophosphamide in overcoming a suppressor barrier in the adoptive transfer of contact sensitivity. Administration of 14-12 mAb just prior to immunization results in the augmentation of contact sensitivity, antibody and plaque-forming cell responses. These results demonstrate the versatility of this reagent for the study of Ts cell activity. PMID- 2970970 TI - Aggravation of experimental cutaneous leishmaniasis in mice by administration of interleukin 3. AB - Previous studies from our laboratory have shown that some in vitro maintained Leishmania major-specific L3T4+ T cells were capable of exacerbating cutaneous leishmaniasis after adoptive transfer to normal syngeneic mice. Results presented in this report show that these cells released substantial amounts of interleukin 3 (IL 3) and granulocyte-macrophage colony-stimulating factors after specific stimulation in vitro. In order to assess the involvement of such lymphokines in the exacerbation of cutaneous leishmaniasis by these L3T4+ T cells, the effect of the administration of important doses of IL 3 on the course of infection with L. major was investigated. The treatment of genetically susceptible BALB/c mice with IL 3 resulted in an enhancement of the size of lesions and favored the multiplication of parasites at anatomical sites distant from the primary lesion. Although IL 3 did not modify the development of lesions in genetically resistant CBA mice, this lymphokine promoted the growth of Leishmania in lymph node draining the lesion. Finally, the addition of IL 3 to macrophages parasitized in vitro enhanced the survival of intracellular Leishmania major. PMID- 2970971 TI - Involvement in the initiation of T cell responses and structural features of an 85-kDa membrane activation antigen. AB - A monoclonal antibody, 7F7, directed at a recently described membrane activation antigen of 85 kDa was found to inhibit the T cell proliferation induced by an anti-CD3 antibody, phytohemagglutinin A and concanavalin A. The T cell response to allogenic stimulator cells was also weakly inhibited. The inhibition of these T cell responses was only obtained if the antibody was added within the first 8 h (first 24 h in the case of the concanavalin A response) of culture. In addition the antibody inhibited the formation of cellular aggregates seen in stimulated cultures when added within the first 8 h. The membrane glycoprotein recognized by 7F7 is shown to have a slightly different molecular mass on cells of different lineage, a protein core of 55 kDa and to contain 30-50% of N-linked as well as a small amount of O-linked carbohydrates and sialic acid residues. This study suggests that the highly glycosylated membrane activation antigen defined by antibody 7F7 could contribute to the contact between those cells which are involved in the initiation of T cell responses. PMID- 2970972 TI - Expression of CR2/EBV receptors on human thymocytes detected by monoclonal antibodies. AB - The biologic effects of the third component of complement, C3, are mediated via receptors which specifically bind the enzymatic degradation products resulting from the cleavage of C3. One of the products, C3d, has been associated with binding to the second complement receptor CR2 (CD21). This receptor, which is identical to the receptor for Epstein-Barr virus (EBV), has been primarily found on cells of the B lineage, but not on mature T cells or other cells of erythroid or myeloid lineages. In the present investigation, we report the presence of CR2 on human thymocytes. Indirect immunofluorescence analysis employing monoclonal anti-CR2 antibodies revealed a range of thymocyte reactivity from 15% to 63% in thirteen experiments using cells of different donors. Reactivity was always greater with the monoclonal anti-CR2 (CD21) antibody HB-5 than with two other antibodies which recognize distinct epitopes on the CR2 molecule. Two-color immunofluorescence analysis indicated that the brightest of the HB-5-stained thymocytes also reacted with the monoclonal anti-CD1 antibody T6 (immature thymocyte marker) while some of the duller HB-5-staining cells reacted with the monoclonal anti-CD3 antibody Leu-4 (mature thymocyte marker). Immunoprecipitation of CR2 on thymocytes with antibody HB-5 and polyacrylamide gel electrophoretic analysis revealed a protein of 145 kDa molecular mass which is consistent with the size of CR2 found on B lymphocytes. These findings raise several questions regarding the biologic role of CR2-EBV receptor on cells of the T lineage. PMID- 2970973 TI - 5-HT1A, 5-HT1B and 5-HT2 receptor agonists induce differential behavioral responses in neonatal rat pups. AB - Sprague-Dawley rat pups at 3-4 days prenatally were tested in both the absence and presence of milk following administration of various doses of either the 5 HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OHDPAT), the 5-HT1B agonist 1-(3-chlorophenyl)piperazine (mCPP), or the 5-HT2 agonist 1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane (DOI). Administration of 8-OHDPAT decreased mouthing, increased probing and increased behavioral activation. Conversely, the 5-HT2 agonist DOI and the 5-HT1B agonist mCPP increased mouthing and decreased probing. mCPP and DOI differed in their effects on behavioral activation, with mCPP decreasing and DOI increasing this composite behavioral score. mCPP increased grooming, whereas DOI elicited a characteristic unusual positioning of the limbs. Thus it appears that 5-HT1A, 5-HT1B and 5-HT2 receptor subtypes are present in the neonate and elicit differential behavioral responses upon stimulation with selective agonists. Ontogenetic variations in the balance among these receptor subtypes during development may be related to the ontogenetic reversal that has been previously reported in the impact of serotonin manipulations on mouthing and suckling behavior during the neonatal to weanling age period. PMID- 2970974 TI - Evidence that the hyperphagic response to 8-OH-DPAT is mediated by 5-HT1A receptors. AB - The 5-HT1A agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT) at a dose of 1 mg/kg s.c. increased food intake in free feeding rats. 8-OH-DPAT-induced feeding was blocked by metergoline which has comparable affinity for 5-HT1A, 5 HT1B, 5-HT1C and 5-HT2 receptors. This is consistent with the hyperphagia being mediated by an action at 5-HT receptors. Evidence against the involvement of 5 HT2 or 5-HT3 receptors was provided by the lack of effect of methysergide, ketanserin, MDL 72222 and ICS 205930 on the feeding response. Blockade of the hyperphagia by (-)- but not (+)pendolol which stereoselectively interacts with 5 HT1 receptors indicated an involvement of this receptor type. The lack of effect of ketanserin suggests that the 5-HT1C site is not involved as it has high affinity for both 5-HT2 and 5-HT1C receptors. Blockade of the hyperphagia by spiperone suggests mediation by 5-HT1A rather than 5-HT1B receptors. Although spiperone also blocks dopamine and alpha 2-adrenoreceptors, involvement of these sites is unlikely as neither the DA antagonist haloperidol nor the alpha 2 adrenoceptor antagonist idazoxan blocked 8-OH-DPAT-induced feeding. These results indicate that 8-OH-DPAT-induced feeding is mediated by 5-HT1A receptors. PMID- 2970976 TI - Vasorelaxant effects of atrial peptide II on isolated human pulmonary muscle preparations. AB - Isolated human pulmonary arterial preparations were approximately 10-fold more sensitive to atrial peptide II than venous preparations (pD2 values: 7.11 +/- 0.14 and 6.17 +/- 0.14, respectively). Atrial peptide II did not relax isolated human bronchial muscle preparations (pD2 value: greater than 5.5). These data demonstrate that atrial peptide II is a selective relaxant agent in isolated human pulmonary vascular muscle preparations with no effect on isolated human airway muscle preparations. PMID- 2970975 TI - A highly potent and selective H3 agonist relaxes rabbit middle cerebral artery, in vitro. AB - An H3-agonist, R alpha methylhistamine strongly relaxed isolated and perfused rabbit middle cerebral artery precontracted with K+. This vasodilatation was not antagonized by either mepyramine or cimetidine but was competitively antagonized by an H3-antagonist, a mixture of impromidine and cimetidine. Histamine activated the H3-sites in this artery since its concentration-response curve (obtained with mepyramine and cimetidine) was parallel to that of R alpha methylhistamine. Our data indicate that H3-sites could exist in the rabbit cerebral artery. PMID- 2970977 TI - Recovery of muscle from tetrodotoxin-induced disuse and the influence of daily exercise. 2. Muscle enzymes and fatigue characteristics. AB - The fatigue characteristics and the activities of oxidative and glycolytic enzymes were determined in tetrodotoxin (TTX)-induced disuse muscles and in muscles of animals recovering from TTX-induced disuse (TTX-rec). In addition, the effects of additional daily exercise (grid-climbing and swimming) on the fatigue and metabolic profiles of muscles from TTX-rec and control animals were investigated. The activities of citrate synthase (CS), phosphofructokinase (PFK), and alpha-glycerophosphate dehydrogenase (alpha-GPD) were depressed following 28 days of inactivity produced by the chronic neural application of TTX. The response of these muscles to a pattern of stimulation that has been used to classify fast-twitch motor units according to their fatigability (6) (330 ms, 40 Hz, l/s, 4 min) was not affected to any great extent by inactivity, except for a loss in the ability to summate or maintain forces during each 330-ms burst, as fatigue developed. After 28 days of recovery, the concentration of CS had returned to normal, whereas the concentrations of PFK and alpha-GPD remained depressed. TTX-rec muscles, on the other hand, appeared more resistant to fatigue than control muscles, based on several indices of muscle fatigue. Control and TTX muscles responded similarly to daily training. Swimming but not climbing increased the activity of CS and the fatigue resistance of the muscle. Neither exercise influenced the activity of PFK and alpha-GPD. Although the activity of CS was influenced by the level of neuromuscular usage, the former did not appear to play a dominant role in determining the fatigue resistance of the muscle, emphasizing the need to consider other factors as primary determinants of muscle fatigue. PMID- 2970978 TI - Symbioramide, a novel Ca2+-ATPase activator from the cultured dinoflagellate Symbiodinium sp. AB - A novel sphingosine derivative, symbioramide, has been isolated from the laboratory-cultured dinoflagellate Symbiodinium sp. as a sarcoplasmic reticulum (SR)Ca2+-ATPase activator, and its structure elucidated to be 1 on the basis of spectral and chemical means. PMID- 2970979 TI - [Mechanism of the effect of atriopeptin on regional blood flow]. AB - Atriopeptin exerts a direct regulatory vasodilating effect on the mechanism of the regional blood flow maintenance. Under preliminary activation of vascular tone the atrial factor can produce the smooth muscle relaxation. Atriopeptin II is the most active among the three types of atriopeptin, atriopeptin III is the least active. PMID- 2970980 TI - Undersulfation of glomerular basement membrane heparan sulfate in experimental diabetes and lack of correction with aldose reductase inhibition. AB - In this study we examined the effect of experimental diabetes and of treatment with an aldose reductase inhibitor on the level of sulfation of glomerular basement membrane (GBM) heparan sulfate, the principal glycosaminoglycan in this extracellular matrix. Glycosaminoglycans were isolated from GBM purified from control, streptozocin-induced diabetic, and sorbinil-treated diabetic rats and analyzed for sulfate and uronate content. Glomerular yields from diabetic kidneys were greater than those from control animals, but the amount of sulfate per glomerulus in diabetic samples, both untreated and sorbinil treated, did not differ significantly from that in control samples. However, the sulfate-to uronate ratio in heparan sulfate isolated from diabetic GBM (0.34 +/- 0.08) was significantly less than in control samples (0.69 +/- 0.11), and treatment of diabetic rats with an aldose reductase inhibitor did not correct this reduced ratio (0.36 +/- 0.06). The results indicate that there is an undersulfation of heparan sulfate of GBM in experimental diabetes, an abnormality that may contribute to loss of anionic sites and decreased charge selectivity of the glomerular filtration barrier. The findings further suggest that this abnormality results from disturbances in glycosaminoglycan synthesis and/or metabolism in diabetes that are independent of polyol-pathway activation in the renal glomerulus. PMID- 2970981 TI - Effect of insulin therapy on established diabetic nephropathy in rats. AB - The effectiveness of insulin therapy on early diabetic nephropathy has not been established. In this study we examined the influence of continuous subcutaneous insulin on the progression of established nephropathy in streptozocin-induced diabetic rats. Normal controls and diabetic rats were studied for 11 mo. During the first 6 mo, all the diabetic rats received 2 U protamine zinc insulin s.c. twice weekly. During the last 5 mo of study, diabetic rats either continued on the occasional subcutaneous insulin regimen or received regular insulin by continuous subcutaneous infusion. Six months after the initiation of the study, the diabetic rats were severely hyperglycemic, and their relative mesangial areas had increased. Continued poor glycemic control in the rats receiving occasional insulin was associated with relative increased mesangial area (25.2 +/- 1.0% of glomerular area) and significant proteinuria (148 +/- 17 mg/24 h) compared with normal controls. In contrast, the use of continuous subcutaneous insulin therapy with improved glycemic control arrested mesangial changes (19.5 +/- 1.4% of glomerular area) and prevented the excessive proteinuria (71 +/- 13 mg/24 h). Indeed, these parameters did not differ from age-matched controls. We conclude that in the rat, continuous subcutaneous insulin therapy instituted after the development of early glomerular pathology is effective in arresting the disease process. PMID- 2970982 TI - Effect of L-carnitine treatment on lipid metabolism and cardiac performance in chronically diabetic rats. AB - The beneficial effects of L-carnitine administration were studied in vivo in isolated perfused working hearts from control and diabetic rats. Control and streptozocin-induced diabetic (STZ-D) rats were treated daily for 6 wk with high dose L-carnitine (3 g.kg-1.day-1 i.p.). STZ-D results in loss of body weight and hypoinsulinemia. These effects were not altered by L-carnitine treatment. Myocardial free-carnitine levels were decreased in the untreated diabetic rats. L Carnitine treatment of the diabetic rats increased myocardial free-carnitine levels, which were comparable with those of control rats. Six weeks after STZ administration, hearts from untreated diabetic animals exhibited depressed left ventricular developed pressure, cardiac contractility, and ventricular relaxation rates compared with control animals. However, this depression was not seen in the L-carnitine-treated diabetic animals. L-Carnitine treatment of diabetic rats significantly reduced plasma glucose and lipid levels but had no effect on control rats. Furthermore, thyroid hormone levels were higher in the L-carnitine treated diabetic rats than in the untreated diabetic group. The data suggest that high-dose L-carnitine treatment may reduce the severity of diabetes and result in improved cardiac performance. PMID- 2970983 TI - Expression of class II MHC antigens in murine pancreas after streptozocin-induced insulitis. AB - In this study, most class II antigen expression during streptozocin-induced insulitis was associated with the mononuclear cells infiltrating the pancreas. During the early stages of the insulitis response, phagocytic cells expressing class II antigens and containing beta-cell debris were often observed. Although there was de novo expression of class II antigens by ductular epithelium associated with the exocrine portion of the pancreas and transient expression by vascular endothelial cells, the endocrine cells of the islets remained devoid of class II antigens throughout the period of observation (42 days). PMID- 2970985 TI - [Visible control of peritoneal perforation in surgical pelviscopy]. AB - From the inception of laparoscopy, the primary blind incision has presented an as yet unresolved risk. In "open laparoscopy", the abdomen is opened in the classic manner as though a "mini-laparotomy" were being performed. This is time consuming, cosmetically unsatisfactory, and restricted locally. Using a newly tried-out technique, and adhering to the classic transumbilical Z-incision technique, both the muscle layer and the peritoneum beneath it are punctured under endoscopic sight. Here, the elliptically ground end of a trocar capsule slides along the epiperitoneum, until light shining through allows a growth-free site to be diagnosed. The preceding inflation of the pneuperitoneum is performed without risk by use of aspiration tests, vacuum tests, and sound tests, and simultaneous measurement of insufflation and static pressure (Semm monofil bivalent system). PMID- 2970984 TI - Adenosine triphosphatase in nerves and ganglia of rats with streptozotocin induced diabetes or galactosaemia; effects of aldose reductase inhibition. AB - This study measured the ouabain-sensitive and ouabain-resistant adenosine triphosphatase activity in homogenates of the sciatic nerves and of pooled fourth and fifth lumbar dorsal root ganglia from rats fed 20% galactose or made diabetic with streptozotocin for either 4 or 8 weeks. Diabetes caused reductions in both fractions of sciatic nerve adenosine triphosphatase activity. After 8 weeks the ouabain-sensitive fraction was 54% of control (p less than 0.05) and the ouabain resistant fraction was 57% of control (p less than 0.05). Galactose feeding more than doubled the ouabain-sensitive adenosine triphosphatase activity in the sciatic nerve (225% of control after 4 weeks, 215% of control after 8 weeks of galactose feeding, both p less than 0.01) and produced a progressive increase in the ouabain-resistant fraction (119% of control at 4 weeks (p less than 0.05) and 176% of control at 8 weeks (p less than 0.01)). In a group of rats fed galactose for 5 days, sciatic nerve ouabain-sensitive adenosine triphosphatase activity was 165% of control. Treatment with the aldose-reductase inhibitors tolrestat, ponalrestat or sorbinil prevented accumulation of polyol and depletion of myo inositol in the sciatic nerves, indicating effective inhibition of aldose reductase. These drugs prevented completely the effect of galactose on the sciatic nerve adenosine triphosphatase activity, but had no significant effect on the reduction in adenosine triphosphatase activity in the sciatic nerves of diabetic rats. In the dorsal root ganglia galactose feeding had no measurable effect on the adenosine triphosphatase activity. Diabetes caused a modest numerical reduction in the ouabain-sensitive activity only.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2970986 TI - Medroxyprogesterone acetate (MPA) enhances liver NADPH-generating enzyme activities in normal rats. AB - The aim of the present study was to evaluate the effects of medroxyprogesterone acetate (MPA), an inducer of liver drug metabolism, on the ability of liver to generate NADPH, a reducing cofactor for drug oxidation reactions in normal rats and to compare these results with those obtained in rats receiving phenobarbital (PB), a well known inducer of liver drug metabolism. The results showed that: 1. Administration of MPA (100 mg/kg body wt) for a week increased liver wt and NADPH cytochrome P-450 reductase activity, suggesting that the compound induced liver drug metabolism. 2. The regimen also increased the activities of two NADPH generating enzymes, isocitrate dehydrogenase and malic enzyme, suggesting that MPA enhanced the capacity of normal liver tissue to produce NADPH. 3. Phenobarbital treatment increased the activities of three NADPH generating enzymes, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and malic enzyme, suggesting that MPA and PB differ in their effects on the liver NADPH-producing system. PMID- 2970987 TI - Microsomal effects of cyproterone acetate and flutamide in rat testis. AB - 1. Effects of two doses of cyproterone acetate (CA) and flutamide (FLU) (50 and 100 mg/kg/3 days) on the testicular steroidogenesis in male rats have been investigated, by measuring the content of cytochrome P-450, cytochrome b5 and cytochrome c reductase activity in the microsomal fraction. 2. CA provoked a significant decrease in cytochrome P-450 content while FLU induced an increase with the lowest dose but a significant decrease after administration of 100 mg/kg. 3. On the other hand, in all cases the cytochrome b5 and cytochrome c reductase activity remained unchanged. 4. The plasma levels of LH and testosterone were measured after CA and FLU injection (50 and 100 mg/kg/3 days). 5. CA administration provoked a reduction in blood levels of these hormones while FLU induced a significant increase in both. 6. These data suggested that CA and FLU modified the cytochrome P-450 in rat testes by an indirect mechanism, probably through the modification of LH plasma levels. PMID- 2970988 TI - The human factor VII gene is polymorphic due to variation in repeat copy number in a minisatellite. AB - The gene coding for human factor VII, a vitamin K-dependent coagulation factor, contains five minisatellite imperfect tandem repeats with monomer element lengths ranging from 14 to 37 bp, and copy numbers ranging from 6 to 52. Three of these repeats are entirely within introns, one is entirely in an untranslated portion of an exon, and one spans an exon-intron border and contains coding sequence. A consensus sequence derived from a comparison of the monomers is similar to a core sequence found in other minisatellites. All of the minisatellites display higher order periodicities. At least one of these minisatellites is polymorphic. A variation in repeat copy number has been observed in a tandem-repeat region in the seventh factor-VII intron. PMID- 2970989 TI - Isolation and characterization of the Bos taurus beta-casein gene. AB - The expression of casein genes in the mammary cells is regulated by peptide and steroid hormones. To investigate the controlling mechanisms we have isolated and characterized the bovine beta-casein gene. The gene has the size of 8.6 kb, which is 7.8 times longer than the corresponding mRNA composed of nine exons. The genomic clones include additional 8.5-kb and 4.5-kb sequences of the 5'- and 3' flanking regions. We have determined the sequences of the 5' and 3' ends of the gene and compared them with the respective sequences of the rat beta-casein gene. Conserved sequences are identical or homologous to the potential binding sites for nuclear factors and for glucocorticoid and progesterone receptors. The regulatory region contains two different TATA signals and a repeat sequence between them. PMID- 2970990 TI - Structure and characterisation of a duplicated human alpha 1 acid glycoprotein gene. AB - Human alpha 1-acid glycoprotein (AGP), also known as orosomucoid, is a major acute-phase plasma protein. The amino acid sequence of AGP, which was determined by sequencing from protein isolated from pooled plasma, contained amino acid substitutions in 21 different positions. Genomic and cDNA clones which correspond to one of the possible amino acid sequences have been previously reported. In this paper we present the complete nucleotide sequence of a second gene, AGP2 which is located approx. 3.3 kb downstream from AGP1. The derived amino acid sequence of AGP2 contains 19 of the possible alternative amino acid substitutions as well as two additional differences. It is clear from the results presented here that the AGP in human plasma is the product of two separate gene loci. PMID- 2970991 TI - [Physiological and hygienic requirements for the rational construction of a tractor operator's seat]. PMID- 2970992 TI - [Mathematical approach to the evaluation of the immunological status of allergy patients]. PMID- 2970993 TI - [Adaptive reactions in workers in the metallurgical industry]. PMID- 2970994 TI - [Data on the substantiation of the maximum permissible level of dust of Viskoza 77 viscose fiber in the air of the work area]. PMID- 2970995 TI - [The main approaches to improving working conditions in the production of bronze and brass alloys]. PMID- 2970996 TI - [Occupational diseases among physicians]. PMID- 2970997 TI - [Employment of invalids in agriculture and evaluation of its expediency]. PMID- 2970998 TI - [Effect of artificial ozone treatment on microbial air contamination in the textile industry]. PMID- 2970999 TI - [Employment and medical rehabilitation in diseases caused by physical stress]. PMID- 2971000 TI - [Experience in using rooms for psycho-emotional relaxation at an engineering plant]. PMID- 2971001 TI - [Characteristics of the structure of occupational pathology of the skin in recent years]. PMID- 2971002 TI - [Occupational hygiene and the health measures at present-day coal-processing plants]. PMID- 2971004 TI - Behavioural objectives model in nursing curriculum design and development: its problems and the alternative approach. PMID- 2971003 TI - The effects of host sex and sex hormones on Trichinella spiralis Owen, 1835 and T. pseudospiralis Garkavi, 1972 in the mouse. AB - The intensity of Trichinella spiralis and T. pseudospiralis infections and effects of sex hormones, oestrogen (Agofollin) and testosterone (Agovirin), were studied in SPF ICR mice with respect to different sexes of the host. Males were found to be more sensitive to T. spiralis and T. pseudospiralis infections than the females. The number of larvae in the infected mice decreased after the application of the female sex hormone oestrogen to males and male sex hormone testosterone to females. PMID- 2971005 TI - Group processes in nursing education. PMID- 2971006 TI - The function of role play in nursing education with reference to adult learning. PMID- 2971007 TI - Classroom knowledge and its application to clinical practice. PMID- 2971008 TI - The developing role of the nurse. PMID- 2971009 TI - T3 reverses the changes in met-enkephalin and beta-endorphin contents in the anterior lobe, but not the neuro-intermediate lobe of the pituitary of rats rendered hypothyroid by PTU-treatment. AB - The changes in met-enkephalin and beta-endorphin contents in the pituitary in PTU induced hypothyroidism were studied in the rat. After 2 weeks of PTU-treatment, both IR-met-enkephalin and IR-beta-endorphin contents in the pituitary were significantly reduced. Gel filtration chromatography followed by radioimmunoassay showed that the immunoactivities in the peaks of precursors, met-enkephalin, beta lipotropin and beta-endorphin were all lower in the pituitaries from the PTU treated rats. In another experiment, some of the PTU-treated rats were injected daily with 500 micrograms T3/kg b.w. In the hypothyroid rats, IR-met-enkephalin and IR-beta-endorphin contents were decreased in both the anterior and neurointermediate lobes. Only the changes in the anterior lobe were reversed by T3 treatment. In conclusion, while the effects on the anterior lobe are probably due to a deficiency in thyroid hormones, the mechanism for the decrease of opioid peptide contents in the neurointermediate lobe is still unclear. PMID- 2971010 TI - Absent B-endorphin response to clonidine in obese children. AB - Plasma B-Endorphin (B-EP), Growth Hormone (GH) and cortisol response to 100 mcg/m2 b.s., i.v. clonidine (an alpha 2-adrenergic agonist) were evaluated in 17 normal weight children (8 prepubertal and 9 pubertal) and in 15 children with simple exogenous obesity (7 prepubertal and 8 pubertal, weight excess ranging from 29% to 97%). All the hormones were measured by radioimmunoassay either directly in the plasma (GH and cortisol) or after extraction and chromatography (B-EP). Obese prepubertal and pubertal children showed basal B-EP levels significantly higher than in controls and no differences were found in GH and cortisol levels. While in controls clonidine stimulated a significant release of plasma GH and B-EP in obese patients, irrespective of pubertal development, no changes were found. Cortisol levels decreased in both groups. These data suggest an impaired adrenergic control of GH and B-EP secretion in children with simple exogenous obesity. PMID- 2971011 TI - In vivo and in vitro studies in a 46, XY phenotypically female infant with 17 ketosteroid reductase deficiency. AB - A 46, XY phenotypically female infant with 17-ketosteroid reductase (17-KSR) showed normal plasma androgens for chromosomal sex shortly after birth, but did not show the physiologic testosterone rise. One intramuscular injection with human chorionic gonadotropin resulted in high ratios between androstenedione/testosterone and dehydroepiandrosterone/delta 5-androstenediol, confirming the diagnosis. In spermatic vein plasma similarly elevated ratios were found. A urinary steroid profile revealed elevated levels of metabolites of 17-OH progesterone and androstenedione. In vitro studies in testicular tissue showed a decreased capacity of 17-ketosteroid reductase, the reduction capacity being more affected than the oxidation capacity. The activity of 3 beta-hydroxysteroid dehydrogenase was slightly increased. The serial analysis of plasma androgens provides more insight in the natural history of 17-ketosteroid reductase. PMID- 2971012 TI - Evidence of immunoreactive human atrial natriuretic peptides in human anterior pituitary cells. PMID- 2971013 TI - Activation of murine Kupffer cell tumoricidal activity by liposomes containing lipophilic muramyl dipeptide. AB - The ability of liposomes containing a lipophilic muramyl dipeptide, N acetylmuramyl-L-alanyl-D-isoglutamine-glycerol dipalmitate, to induce Kupffer cell tumoricidal activity has been investigated. Liposomal N-acetylmuramyl-L alanyl-D-isoglutamine-glycerol dipalmitate was 16-fold more potent than liposomal N-acetylmuramyl-L-alanyl-D-isoglutamine and 2,400-fold more potent than N acetylmuramyl-L-alanyl-D-isoglutamine in inducing Kupffer cell tumoricidal activity in vitro. A single i.v. injection of liposomes containing N acetylmuramyl-L-alanyl-D-isoglutamine-glycerol dipalmitate was capable of inducing Kupffer cell tumoricidal activity as measured against B16-melanoma cells after Kupffer cell isolation. Maximal cytotoxic activity was obtained with 1 microgram muramyl dipeptide-glycerol dipalmitate encapsulated within liposomes: doses of 10 or 100 micrograms inhibited tumoricidal activity. Kupffer cells from mice treated with liposomes containing N-acetylmuramyl-L-alanyl-D-isoglutamine glycerol dipalmitate remained cytotoxic for at least 6 days after injection. Liposomal N-acetylmuramyl-L-alanyl-D-isoglutamine was significantly less potent than liposomal N-acetylmuramyl-L-alanyl-D-isoglutamine-glycerol dipalmitate in inducing Kupffer cell tumoricidal activity in situ. N-Acetylmuramyl-L-alanyl-D isoglutamine was capable of inducing Kupffer cell tumoricidal activity in vitro: its failure to induce tumoricidal activity in situ at doses of 1,000 micrograms demonstrates the utility of liposomal carriers for the in vivo activation of Kupffer cells by muramyl dipeptides. PMID- 2971014 TI - Activation of liver macrophages following phenobarbital treatment of rats. AB - Phenobarbital is a potent inducer of hepatic cytochrome P-450 and is a tumor promoter in the two-stage model of liver carcinogenesis. In the present studies, we show that phenobarbital also induces an accumulation of activated macrophages in the livers of treated rats. These macrophages are larger and more stellate than resident Kupffer cells and are highly vacuolated. In addition, macrophages isolated from livers of phenobarbital-treated rats display increased phagocytosis of sheep red blood cells, chemotaxis toward the complement fragment C5a and enhanced production of hydrogen peroxide. Biologically active mediators released by activated macrophages have been implicated in tumor promotion as well as in the regulation of cytochrome P-450-mediated drug biotransformation. We propose that the activated macrophages that accumulate in the liver following phenobarbital treatment may contribute, at least in part, to the biological responses to this drug. PMID- 2971015 TI - Peripheral arterial vasodilation hypothesis: a proposal for the initiation of renal sodium and water retention in cirrhosis. AB - Renal sodium and water retention and plasma volume expansion have been shown to precede ascites formation in experimental cirrhosis. The classical "underfilling" theory, in which ascites formation causes hypovolemia and initiates secondary renal sodium and water retention, thus seems unlikely. While the occurrence of primary renal sodium and water retention and plasma volume expansion prior to ascites formation favors the "overflow" hypothesis, the stimulation of the renin angiotensin-aldosterone system, vasopressin release and sympathetic nervous system associated with cirrhosis is not consonant with primary volume expansion. In this present article, the "Peripheral Arterial Vasodilation Hypothesis" is proposed as the initiator of sodium and water retention in cirrhosis. Peripheral arterial vasodilation is one of the earliest observations in the cirrhotic patient and experimental animals with cirrhosis. Arterial vasodilators and arteriovenous fistula are other examples in which renal sodium and water retention occur secondary to a decreased filling of the arterial vascular tree. An increase in cardiac output and hormonal stimulation are common features of cirrhosis, arteriovenous fistula and drug-induced peripheral arterial vasodilation. However, a predilection for the retained sodium and water to transudate into the abdominal cavity occurs with cirrhosis because of the presence of portal hypertension. The Peripheral Arterial Vasodilation Hypothesis also explains the continuum from compensated to decompensated cirrhosis to the hepatorenal syndrome. PMID- 2971016 TI - The M2 autoantigens of primary biliary cirrhosis are not subunits of the mitochondrial H+-ATPase. PMID- 2971017 TI - Immunophenotypes of malignant lymphoma centroblastic-centrocytic and malignant lymphoma centrocytic: an immunohistologic study indicating a derivation from different stages of B cell differentiation. AB - Five cases of intermediate lymphocytic lymphoma (ILL), 13 cases of malignant lymphoma centrocytic (MLCC), and 27 cases of malignant lymphoma centroblastic centrocytic (MLCBCC) were studied morphologically and with the aid of a panel of monoclonal antibodies. The immunophenotypes of ILL and MLCC (IgM+/IgD+, MT1+, CALLA-) indicate a mantle zone or very early follicle center derivation. The immunophenotypes of MLCBCC (IgM+ or IgG/IgA+, MT1-, CALLA+) indicate a "true" follicle center derivation. The morphologic diversity of MLCBCC could not be related to specific immunophenotypes. PMID- 2971018 TI - Maternal levels of pregnancy-specific beta 1-glycoprotein (SP-1) are elevated in pregnancies affected by Down's syndrome. AB - Concentrations of pregnancy-specific beta 1-glycoprotein (SP-1) were measured in maternal blood and amniotic fluid of patients with a trisomic fetus and compared with that of a cytogenetically normal fetus at weeks 16-19 of pregnancy. The SP-1 concentrations were significantly elevated in the sera of women with a Down's syndrome fetus, whereas amniotic fluid levels were only slightly increased. It is suggested that high levels of maternal SP-1 in the second trimester of pregnancy may be a valuable indicator in the prenatal detection of fetal trisomy 21. PMID- 2971019 TI - [Porokeratosis and immunosuppressive therapy. Presentation of a further case and review of the literature]. PMID- 2971020 TI - Streptococcus mutans gtfA gene specifies sucrose phosphorylase. AB - The ethanol-insoluble product formed from sucrose by purified enzyme encoded by the gtfA gene from Streptococcus mutans, expressed in recombinant Escherichia coli, was analyzed by 13C and 31P nuclear magnetic resonance. The product was identified as alpha-D-glucose-1-phosphate, and it was concluded that the GTF-A enzyme is sucrose phosphorylase (sucrose:orthophosphate alpha-D glucosyltransferase [EC 2.4.1.7]). PMID- 2971021 TI - Activation of mononuclear cells to be used for hybrid monoclonal antibody-induced lysis of human ovarian carcinoma cells. AB - Recently we reported that cytotoxic T-cell clones can be retargeted to unrelated tumor cells by bispecific monoclonal antibodies (MAbs), anti-CD3 and anti-ovarian carcinoma (alpha OC/TR) (Mezzanzanica et al., 1988). In the perspective of in vivo tumor immunotherapy, as an alternative to cytotoxic T lymphocytes (CTL) from T-cell clones, since human peripheral blood mononuclear cells (PBMCs) without stimulation were quite ineffective, a suitable in vitro activation method was developed to render PBMCs lytic for relevant targets in the presence of the bispecific hybrid MAb alpha OC/TR. This activation protocol was applied to PBMCs from 9 healthy donors (HD) and 6 ovarian carcinoma patients (P) and to tumor associated lymphocytes (TAL) from 4 ovarian carcinoma P. The method consisted of in vitro stimulation with phytohemagglutinin (PHA) for 2 days, followed by culture with a low dose of recombinant human interleukin-2 (rIL-2) for 6 to 10 days. The antibody-mediated lysis of CTL from HD PBMCs was found to be specifically directed against cells expressing the relevant ovarian tumor antigen when different tumor cell lines and short-term cultures of tumor and normal cells were tested. The antibody-mediated lysis of CTL from P PBMCs or TAL was efficient both on autologous and allogeneic ovarian tumor cells, whereas no reactivity with autologous normal cells was observed and LAK activity was only evident in 1 out of 4 cases. The hybrid antibody induced cytotoxic activity of CTL from P was, however, lower than that of CTL from HD. PMID- 2971022 TI - Endocrine effects of a trickle infection with Dictyocaulus viviparus in the calf. PMID- 2971023 TI - A diurnal rhythm in opsin content of Rana pipiens rod inner segments. AB - Quantitative electron microscope immunocytochemistry, employing an antibody specific to opsin, was used to evaluate the amount and location of opsin in Rana pipiens rod photoreceptors throughout a 24 hr light/dark cycle. We found a distinct diurnal rhythm in the density of anti-opsin labeling of the rough endoplasmic reticulum (RER) and Golgi apparatus in the myoid region of the rod inner segment. Opsin labeling of these organelles was lowest at light onset, increasing thereafter by three- to four-fold, and remained high until 2 hr into the dark phase. A fall in labeling density occurred within the following 4 hr, and remained low for the remainder of the dark phase. Our finding of a diurnal rhythm regulating inner segment opsin transport in Rana pipiens contrasts with published observations on outer segment membrane turnover, since it has been shown that the rates of disc formation and disc shedding are governed by environmental lighting alone in this species. These results imply that there is opsin pooling in the inner segment during the first 14 hr of a 24 hr light/dark cycle; thereafter the loss of inner segment opsin due to mobilization of this protein from the Golgi exceeds the rate of formation of new opsin. There was no evidence of accumulation of opsin-containing vesicles near the cilium or in the ellipsoid just prior to light onset. At light onset, prominent opsin labeling was identified at the proximal portion of the outer segment in regions separate from the disc stack. In two separate experiments, additional groups of frogs were killed around the time of light onset and were examined by conventional transmission electron microscopy. Disordered disc membranes were seen at the base of the outer segment which were not in register with the disc stack. These disordered membranes were observed as early as 2 hr before light onset, and were no longer observed by 1 hr after light onset. We suggest that these disordered membranes reflect a step in the biogenesis of new discs, serving as a pool of membrane that forms during the later part of the dark cycle. It appears that light onset triggers the ordering of neatly registered discs from this new membrane, rather than assembly of new membrane from pooled transport vesicles in the inner segment. PMID- 2971024 TI - Immunoanalysis of keratan sulfate proteoglycan from corneal scars. AB - Corneal keratan sulfate proteoglycan (KSPG) from scar tissue of experimental penetrating corneal wounds in rabbits was analyzed 2-8 weeks after injury using three previously characterized antibodies. Keratan sulfate (KS) was identified in 2 week scars and normal corneal tissue by indirect immunofluorescence using a monoclonal antibody against sulfated KS epitopes. KSPG was measured in unfractionated extracts of scar and of normal corneal tissue using a "sandwich" enzyme-linked immunosorbent assay (ELISA). In extracts of 2 week scars, KSPG molecules reacting with two different anti-KS monoclonal antibodies were 55% and 82% as abundant as in normal tissue extracts. Ion exchange high performance liquid chromatography (HPLC) of tissue extracts found qualitatively similar elution profiles of KSPG antigens from both scar and normal tissues. Direct ELISA of the HPLC-purified KSPG showed identical quantitative binding of antibodies against core protein and KS from normal and scar tissue. KS in the HPLC-purified extracts was sensitive to digestion with endo-beta-galactosidase, whereas core protein antigens were not affected by this enzyme, as expected. Alteration of the antigenic characteristics of the KSPG of scars was detected with a competitive immunoassay using immobilized monoclonal antibodies against KS. KS in extracts from 2, 6, and 8 week scars competed only 5-11% as effectively as KS from normal cornea, although core protein antigens in the scar extracts competed 61-80% as well as those of normal cornea.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971025 TI - Retinal histamine synthesis is increased in experimental diabetes. AB - We examined retinal de novo histamine synthesis mediated by retinal histidine decarboxylase in normal and streptozotocin-diabetic male, Sprague Dawley rats that were diabetic for 21 days. We also examined effects of insulin and alpha hydrazinohistidine (alpha HH) treatments on retinal histamine synthesis in this diabetic model. alpha HH is a specific inhibitor of histidine decarboxylase. Results indicate that the retina contains an active histidine decarboxylase enzyme system, and that in streptozotocin diabetes retinal histamine synthesis is increased 197%. Both insulin and alpha HH independently reverse and normalize retinal histamine synthesis. These data thus indicate that the retinal inducible histamine pool is increased in experimental diabetes, and that insulin is an important modulator of retinal histamine metabolism. This newly described retinal metabolic alteration may be one factor responsible for increased retinal vascular permeability in diabetic retinopathy. PMID- 2971026 TI - In vitro immunological function of human corneal fibroblasts. AB - Gamma-interferon-induced HLA-DR-like antigens on normal human fibroblasts were compared functionally to HLA-DR antigens on peripheral blood mononuclear leucocytes (PBML) from the same individual. Irradiated DR+ fibroblasts did not stimulate allogeneic PBML to divide in a mixed lymphocyte fibroblast culture (MLFC) as determined by 3H-thymidine incorporation, although lymphocytes autologous to the fibroblasts stimulated normally. Co-culture with interleukin 1 (IL-1), IL-2, 2-mercaptoethanol and indomethacin had no effect on MLFC. Pretreatment of stimulator fibroblasts with neuraminidase to modify sialoglycoproteins increased responses only modestly. Both DR+ and DR- fibroblasts inhibited a normal mixed lymphocyte culture (MLC) response in a dose dependent fashion. These results suggest that the induction of HLA-DR-like molecules on fibroblasts is not sufficient to initiate host sensitization and that fibroblasts may inhibit normal lymphoproliferation. PMID- 2971027 TI - Polymorphic markers of human T-cell receptor alpha and beta genes. Family studies and comparison of frequencies in healthy individuals and patients with multiple sclerosis and myasthenia gravis. AB - Polymorphic markers for the human T-cell receptor (TcR) genes are described. With a TcR beta-chain probe, polymorphic allelic fragments of 5.4 and 1.8 kb were detected in KpnI digests, and fragments of 12.5 and 11.5 kb were seen in the BglII digests. Polymorphism in alpha chain genes was observed in TaqI-digested DNA samples with bands at 10.2 and 6.2-2.1 kb. Mendelian codominant inheritance for all three polymorphisms was confirmed in family studies. The gene frequencies for these alleles were determined in a sample of 70 normal unrelated Caucasian individuals, and were shown to be in Hardy-Weinberg equilibrium. There were no significant differences in the frequency of these polymorphic alpha and beta alleles between patients with multiple sclerosis and patients with myasthenia gravis as compared to a panel of control healthy individuals. PMID- 2971028 TI - Additivity of radiosensitization by the combination of SR 2508 (etanidazole) and Ro 03-8799 (pimonidazole) in a murine tumor system. AB - The nitroimidazole radiosensitizers SR 2508 and Ro 03-8799 have different dose limiting toxicities in man and hence can be used in combination. Such therapy will be beneficial only if their radiosensitizing properties are additive, which this study sought to determine using clinically relevant radiosensitizer concentrations in the EMT6 tumor in the flanks of BALB/c mice. 240 mg/kg of each drug gave tumor concentrations (+/- 2 se) 55 min after i.v. administration of the combination of 50.4 +/- 10.6 micrograms/g (236 +/- 50 nmol/g) for SR 2508 and 39.7 +/- 9.0 micrograms/g (137 +/- 31 nmol/g) for Ro 03-8799. The radiosensitization by both agents administered both singly and in combination at 240 mg/kg and singly at 480 mg/kg was measured, giving sensitizers 30 min before 20 Gy of 250 kV X rays. Tumor response was assayed by clonogenic cell survival. SER values (with 95% confidence limits) were 1.28 (1.20-1.37) for 240 mg/kg SR 2508, 1.20 (1.10-1.30) for 240 mg/kg Ro 03-8799, 1.46 (1.33-1.59) for 240 mg/kg of both drugs in combination, 1.46 (1.38-1.55) for 480 mg/kg SR 2508 and 1.46 (1.31-1.62) for 480 mg/kg Ro 03-8799. These data confirm the additivity of radiosensitization by the two drugs administered in combination. PMID- 2971029 TI - Mechanism of action of the selective tumor radiosensitizer nicotinamide. AB - Nicotinamide has been shown to selectively enhance the radiation damage of tumors in preference to normal tissues. Our present study was an investigation into the mechanism responsible for this effect in the SCCVII/St tumor model grown on the backs of C3H/km mice. A large single injection of nicotinamide (1000 mg/kg), given intraperitoneally 60 minutes before whole body irradiation, significantly enhanced the radiation response of SCCVII tumors as measured by an in vivo/in vitro excision assay performed 24 hr following irradiation. It also gave rise to an almost 4-fold reduction in the binding of 14C-misonidazole, injected 1 hr after the nicotinamide and measured by scintillation counting of excised tumor material 24 hr later. This suggested that nicotinamide was decreasing the degree of tumor hypoxia. Attempts were made to correlate these results with nicotinamide induced changes in tumor blood flow using the techniques of 133Xe clearance, 86RbCl extraction and Hoechst 33342 fluorescent labelling. Nicotinamide produced between a 30-40% increase in mean tumor cell fluorescence of Hoechst 33342, which was consistent with an increase in tumor blood flow. A similar response was obtained using the uptake of 86RbCl as the end point. However, no statistically significant difference was seen between the tumor blood flow of control and nicotinamide treated mice using the 133Xe clearance procedure. These results are discussed with respect to their clinical implications. PMID- 2971030 TI - Hypoxic cell radiosensitizers: illusion or elusion? PMID- 2971031 TI - A histological and histochemical study of the cricopharyngeus muscle in man. AB - The human cricopharyngeus muscle was investigated by dissection and by histological, histochemical and morphometric methods. Muscle fibres in the cricopharyngeus were found to be similar in appearance to those of the lateral part of the quadriceps femoris, although they were generally much smaller and more variable in size. The endomysial connective tissue was markedly increased in the cricopharyngeus and muscle spindles were not found. Certain features normally considered to be pathological were also noted in the cricopharyngeus muscles. The fibre type population consisted mainly of histochemically 'slow-twitch' richly oxidative fibres. This finding is consistent with the proposed function of this muscle in its sphincteric role in deglutition, vomiting, eructation and in the control of aerophagia. PMID- 2971032 TI - Microbial transformation of 6-O-methylerythromycin derivatives. AB - Mucor circinelloides f. griseo-cyanus IFO 4563 was found to convert 6-O methylerythromycin A (TE-031, A-56268) to (14R)-14-hydroxy-6-O-methylerythromycin A [14R)-14-hydroxy TE-031). The TLC and spectral data of the conversion product were perfectly identical with those of an active major metabolite of TE-031 in humans (M-5). A related antibiotic, 6-O-methylerythromycin B (TB-010), was able to be similarly transformed to its C-14 hydroxy analogue [14R)-14-hydroxy-6-O methylerythromycin B, (14R)-14-hydroxy TB-010). The MICs of (14R)-14-hydroxy-6-O methylerythromycin B against some Gram-positive bacteria were almost equal to those of 6-O-methylerythromycin B. It is suggested that the hydroxylation at C-14 of 6-O-methylerythromycins A and B scarcely reduces their in vitro activity. PMID- 2971033 TI - 14-Hydroxy-6-O-methylerythromycins A, active metabolites of 6-O methylerythromycin A in human. AB - (14R)-14-Hydroxy-6-O-methylerythromycin A and (14S)-epimer have been isolated as active metabolites from human urine after oral administration of 6-O methylerythromycin A (TE-031, A-56268). The structures of these metabolites were determined by means of mass, 1H and 13C NMR spectroscopies. Antimicrobial activities of (14R)-14-hydroxy-6-O-methylerythromycin A, a major metabolite, were comparable to those of the parent drug TE-031, whereas (14S)-14-hydroxy-6-O methylerythromycin A was 4- to 8-fold less active than TE-031 against bacterial standard strains. PMID- 2971034 TI - Behavioral momentum in the treatment of noncompliance. AB - Behavioral momentum refers to the tendency for behavior to persist following a change in environmental conditions. The greater the rate of reinforcement, the greater the behavioral momentum. The intervention for noncompliance consisted of issuing a sequence of commands with which the subject was very likely to comply (i.e., high-probability commands) immediately prior to issuing a low-probability command. In each of five experiments, the high-probability command sequence resulted in a "momentum" of compliant responding that persisted when a low probability request was issued. Results showed the antecedent high-probability command sequence increased compliance and decreased compliance latency and task duration. "Momentum-like" effects were shown to be distinct from experimenter attention and to depend on the contiguity between the high-probability command sequence and the low-probability command. PMID- 2971035 TI - Carbamazepine and erythema multiforme major. PMID- 2971036 TI - The expression of the 35- and 67-kDa calcimedins is dependent on thyroid hormone. AB - We have investigated the expression of the 35- and 67-kDa calcimedins and calmodulin during fetal and neonatal periods and in adulthood in rat liver, muscle, and brain. The 35- and 67-kDa calcimedin expression in liver and muscle increased during the perinatal period and correlated with the thyroid status of the developing rat. In fact, animals treated with thyroxine demonstrated a precocious appearance of the 35- and 67-kDa calcimedin in liver and muscle. Animals treated with methylthiouracil, an inhibitor of T4 and T3 synthesis, strongly suppressed the synthesis of the calcimedins in these tissues. Neither treatment influenced the levels of either the 35- and 67-kDa calcimedins in brain. In contrast, each tissue examined produced a unique pattern of calmodulin expression during development. None of the tissue calmodulin concentrations changed during hyper- or hypothyroid states. Collectively, these data support the concept that the intracellular calcium signal possesses multiple, independent molecular pathways of mediation. In addition, the variety of these pathways is influenced by hormonal preconditioning in that the cellular response to elevated cytosolic calcium is dependent upon the thyroid status of a tissue. PMID- 2971038 TI - Monoclonal antibodies specific for type 3 protein kinase C recognize distinct domains of protein kinase C and inhibit in vitro functional activity. AB - Monoclonal antibodies (mAbs) which distinguish Type 3 protein kinase C (PKC) from Types 1 and 2 have been obtained from mice immunized with purified Type 3 PKC from rabbit brain cytosol. Most of these mAbs (seven out of eight) selectively recognize Type 3 versus Types 1 and 2 PKC in both enzyme-linked immunosorbent and immunoblot assays. Trypsin treatment of Type 3 PKC reduced the immunoreactivity with 82-kDa PKC and generated immunoreactive fragments of 45 and 35 kDa. The mAbs can be divided into two classes based on their ability to recognize the 45-kDa catalytic fragment (5/8) or the 35 kDa regulatory domain fragment (3/8). Each of the mAbs inhibits phosphorylation of histone or lipocortin by PKC, although the extent of the inhibition varied. Only those mAbs that recognize the 35-kDa regulatory domain inhibited phorbol ester binding. The inhibition of both kinase and binding activities by this group of mAbs was sensitive to the concentration of phospholipid used in the assay. This functional inhibition suggests that these mAbs may be useful for defining the phospholipid binding domain(s) of Type 3 PKC. The mAbs recognized 82-kDa PKC in a variety of cell types; the presence of smaller molecular weight fragments was not consistently found. Distinct immunofluorescence staining patterns were observed with mAbs directed toward different epitopes, suggesting that there may be heterogeneity in the subcellular localization of PKC. The type specificity of these mAbs will make them valuable tools for studying activation and regulation of Type 3 PKC in cell culture model systems. PMID- 2971037 TI - Atrial natriuretic peptide inhibits the agonist-induced increase in extent of myosin light chain phosphorylation in aortic smooth muscle. AB - The effect of atrial natriuretic peptide (ANP) on angiotensin II- and histamine induced contraction and muscle light chain phosphorylation was examined in strips of rabbit aorta smooth muscle. Preincubation of strips with 10(-7) M ANP prior to addition of either agonist inhibits both the increase in extent of myosin light chain phosphorylation and the contractile response to either 5 x 10(-8) M angiotensin II or 10(-5) M histamine without inhibiting the agonist-induced increase in the intracellular free Ca2+ concentration. Furthermore, in muscle strips precontracted with either angiotensin II or histamine, addition of ANP leads to a prompt relaxation and a prompt decrease in the extent of myosin light chain phosphorylation. These data argue that ANP uncouples the initial agonist induced Ca2+ transient from the increase in extent of myosin light chain phosphorylation either by inhibiting the Ca2+-dependent activation of myosin light chain kinase or stimulating the activity of a phosphoprotein phosphatase capable of bringing about the rapid dephosphorylation of phosphorylated myosin light chains. PMID- 2971039 TI - Synthesis and assembly of a catalytically active lysosomal enzyme, beta hexosaminidase B, in a cell-free system. AB - The synthesis and dimerization of beta-chains during the formation of catalytically active beta-hexosaminidase B were studied in a cell-free system. beta-chain mRNA, transcribed from the cloned cDNA with SP6 polymerase, was translated in a rabbit reticulocyte protein-synthesizing system in the presence of dog pancreas microsomal membranes and oxidized glutathione. Under these conditions, the primary beta-chain translation product was translocated into the microsomal vesicles and modified by the addition of N-linked oligosaccharide chains. After transfer into the microsomal vesicles, the beta-polypeptide assumed a conformation resembling the native state as determined by antibody reactivity. Like the authentic precursor enzyme, the microsomally located chains were assembled into dimers and were catalytically active. In intact human fibroblasts, dimerization of beta-chains occurred within 15 min after their synthesis, consistent with a site of assembly in the rough endoplasmic reticulum. The cell free expression system was also useful in establishing the functionality of beta chain initiator methionine codons. By expression of beta-chain mRNAs with altered methionine codons, we demonstrated that polypeptides initiating from any of the first three methionine codons in the beta-chain sequence contain a functional signal sequence and form catalytically active enzymes. PMID- 2971040 TI - Expression of the murine interferon gamma receptor in Xenopus laevis oocytes. AB - This study describes the isolation of mRNA for the murine interferon gamma receptor and its expression in frog oocytes. The binding properties and apparent molecular weight of the murine interferon gamma receptor protein synthesized in frog oocytes is similar to that found on mouse cells. This is the first report of a functional receptor for a polypeptide ligand (interferon gamma) expressed in and directly assayed on frog oocytes. PMID- 2971041 TI - Rat tropoelastin is synthesized from a 3.5-kilobase mRNA. AB - A lambda gt11 cDNA library was constructed from poly(A+) RNA isolated from aortic tissue of neonatal rats and screened with a human tropoelastin cDNA clone. DNA sequence analysis of several overlapping rat clones confirmed the presence of DNA sequences coding for murine tropoelastin and DNA sequences coding for the 3' untranslated region of the rat tropoelastin mRNA. Northern blot analysis of total RNA from aortic tissue of neonatal rats using oligonucleotide probes derived from these rat tropoelastin cDNAs demonstrated the presence of a 3.5-kilobase tropoelastin mRNA. The size of this rat tropoelastin mRNA agrees with previous reports for the size of the mRNA coding for tropoelastin in tissue from several vertebrate species but contrasts with several reports suggesting the presence of a higher molecular weight mRNA species responsible for the synthesis of tropoelastin in rodent tissue. PMID- 2971042 TI - Substitution of gamma Arg-275 by Cys in an abnormal fibrinogen, "fibrinogen Osaka II". Evidence for a unique solitary cystine structure at the mutation site. AB - In an abnormal fibrinogen with impaired fibrin monomer polymerization designed as fibrinogen Osaka II, we have identified substitution of Arg by Cys at position 275 of the gamma chain. This Cys is linked to a free cysteine molecule by a disulfide link as evidenced by fast atom bombardment mass spectrometry. This finding was supported by identification of a single cysteine released from isolated abnormal fragment D1 upon reduction. This unique cystine structure at the mutation site has not been reported heretofore in any abnormal protein including fibrinogen. The substitution may well perturb the structure required for fibrin monomer polymerization, specifically that assigned to the carboxyl terminal D domain of fibrinogen. Indeed, isolated fragment D1 with the Cys substitution failed to inhibit thrombin-mediated clotting of normal fibrinogen and normal fibrin monomer polymerization, while normal fragment D1 inhibited them markedly. Our data seem to provide supporting evidence that the putative polymerization site(s) assigned to the D domain of fibrinogen may be structure dependent, including the carboxyl-terminal segment of the gamma chain as well as a contiguous region that contains the gamma 275 residue. PMID- 2971043 TI - Fatty acids covalently bound to erythrocyte proteins undergo a differential turnover in vivo. AB - Recently, covalently bound fatty acids have been identified on a variety of proteins. Many of these acyl proteins are physiologically important, and the lipid modification often appears to be essential for their function. In this investigation mature erythrocytes have been used to study in detail the metabolic behavior of protein-bound fatty acids. Although deficient in protein synthesis, these cells are still able to covalently attach [3H]palmitic acid to proteins located at the plasma membrane and its associated cytoskeleton. Linkage analyses demonstrated that the labeled polypeptides contained ester- or thioester-bound fatty acids. The covalent binding of fatty acid was rapidly reversible. Half lives of the protein-bound fatty acid molecules ranged from less than 30 min to more than 3 h. The deacylation reaction was not due to a chemically labile linkage of protein and fatty acid but appeared to be physiologically induced. Differences in the fatty acid turnover rates between the acyl proteins suggested an independent regulation of their lipid turnover. A number of proteins underwent dynamic fatty acid acylation, indicating that palmitylated proteins undergoing fatty acid turnover are not a rare exception. PMID- 2971044 TI - Isolation and characterization of the hemichrome-stabilized membrane protein aggregates from sickle erythrocytes. Major site of autologous antibody binding. AB - Because the interaction of denatured hemoglobins (i.e. hemichromes) with the red cell membrane has been associated with several abnormalities commonly observed in hemichrome-containing erythrocytes, we have undertaken to isolate and characterize the hemichrome-rich membrane protein aggregates from sickle cells. The aggregates were isolated by two procedures: one at low ionic strength by centrifugation of detergent-solubilized spectrin-depleted inside-out vesicles, and the other at physiological ionic strength by detergent solubilization of whole cells followed by cytoskeletal disruption and centrifugation. The extensively washed aggregates obtained by both methods yielded similar results. These insoluble complexes were found to be highly cross-linked by predominantly intermolecular disulfide bonds; however, other nonreducible covalent linkages were also observed. Both in the presence and absence of reducing agents, the aggregate disintegrated when the hemichromes were removed by high ionic strength, suggesting that the aggregate depended heavily on the cohesive properties of the hemichromes for stability. Protein assays demonstrated that the aggregates comprised approximately 1.3% of the total membrane protein, roughly two-thirds of which appeared to be globin chains. Other major components identified in the aggregate were band 3, ankyrin, bands 4.1, 4.9, and 5, glycophorins A and B, and autologous IgG. Quantitative analysis of the IgG content demonstrated that three fourths of the surface-bound IgG on washed sickle cells was clustered at these aggregate sites, representing an enrichment of approximately 250-fold over nonaggregated regions of the membrane. Since clustered cell surface IgG is thought to trigger removal of erythrocytes from circulation, the hemichrome induced membrane reorganization at these aggregate sites may be an important cause of the greatly shortened life span of sickle cells. PMID- 2971045 TI - Isolation and characterization of a mutant of L1210 murine leukemia deficient in nitrobenzylthioinosine-insensitive nucleoside transport. AB - L1210 mouse leukemia cells exhibit two distinct types of nucleoside transport activity that have similar kinetic properties and substrate specificity, but differ markedly in their sensitivity to the inhibitor nitrobenzylthioinosine (NBMPR) (Belt, J. A. (1983) Mol. Pharmacol. 24, 479-484). It is not known whether these two transport activities are mediated by a single protein or by separate and distinct nucleoside transport proteins. We have isolated a mutant from the L1210 cell line that has lost the NBMPR-insensitive component of nucleoside transport, but retains NBMPR-sensitive transport. In the parental cell line 20 40% of the nucleoside transport activity is insensitive to 1 microM NBMPR. In the mutant, however, uridine and thymidine transport are almost completely inhibited by NBMPR. Consistent with the loss of NBMPR-insensitive transport, the mutant cells can be protected from the toxic effects of several nucleoside analogs by NBMPR. In contrast, the toxicity of the same analogs in the wild type cells is not significantly affected by NBMPR, presumably due to uptake of the nucleosides via the NBMPR-insensitive transporter. On the other hand, NBMPR-sensitive transport in the mutant appears to be unaltered. The mutant is not resistant to cytotoxic nucleosides in the absence of NBMPR and the cells retain the wild type complement of high affinity binding sites for NBMPR. Furthermore, the affinity of the binding site for the inhibitor is similar to that of parental L1210 cells. These results suggest that NBMPR-sensitive and NBMPR-insensitive nucleoside transport in L1210 cells are mediated by genetically distinct proteins. To our knowledge this is the first report of a mutant deficient in NBMPR-insensitive nucleoside transport. PMID- 2971046 TI - Characterization of an apparently lower molecular weight gamma-chain variant in fibrinogen Kyoto I. The replacement of gamma-asparagine 308 by lysine which causes accelerated cleavage of fragment D1 by plasmin and the generation of a new plasmin cleavage site. AB - Congenitally abnormal fibrinogen Kyoto I with impaired fibrin monomer polymerization contains a normal gamma-chain and a gamma-chain variant (gamma Kyoto I) that has an apparently lower Mr on sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the Laemmli system (Laemmli, U. K. (1970) Nature 227, 680 685) but migrates with apparently normal Mr in the Weber and Osborn system (Weber, K., and Osborn, M. (1969) J. Biol. Chem. 244, 4406-4412). Reverse-phase high performance liquid chromatographic analyses of the cyanogen bromide or lysyl endopeptidase cleavage fragments of the purified gamma-chains of fibrinogen Kyoto I showed the presence of peptides not seen from normal fibrinogen. Amino acid sequence analysis of these peptides indicated that gamma Asn308 of the gamma chain variant is replaced by lysine. Purified fragment D1 of fibrinogen Kyoto I also contains two types of D1 gamma-remnants: normal and apparently lower Mr types. Abnormal fragment D1 is cleaved faster to fragments D2 and D3 by plasmin in the presence of [ethylenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA) than normal fragment D1, as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by immunoblotting using anti-gamma-chain monoclonal antibody. Analysis of peptides released from fragment D1 by plasmin in the presence of EGTA demonstrated the cleavage of the gamma Lys308-Gly309 bond. Fragment D1 of fibrinogen Kyoto I has normal calcium binding properties. The data suggest that a region or conformation containing gamma Asn308 affects the polymerization of fibrin monomers and that the gamma Asn308----Lys replacement causes a conformational change in the gamma-chain which results in the accelerated cleavage of gamma Lys356-Ala357 and gamma Lys302-Phe303 bonds by plasmin and also results in the generation of a new plasmin cleavage site between Lys308 and Gly309 in the presence of EGTA. During these studies, we found that part of the gamma Lys212-Glu213 bond in fragment D1 is cleaved by plasmin in the presence of EGTA. PMID- 2971047 TI - Synthesis of a troponin C cDNA and expression of wild-type and mutant proteins in Escherichia coli. AB - An avian fast striated muscle troponin C cDNA was designed and synthesized from six oligonucleotides using the overlap-fill in method and overproduced in Escherichia coli for the purpose of developing recombinant DNA approaches to study structure-function relationships in this calcium-binding regulatory protein. The recombinant protein isolated from E. coli functions as a bona fide troponin C in all properties that were assayed: calcium binding, calcium dependent conformational change, calcium-dependent interaction with troponin I, and formation of a functional ternary complex with troponin I and troponin T that can confer calcium sensitivity on the actomyosin MgATPase. The initiating methionine was removed by E. coli leaving alanine as the first amino acid, as in the muscle troponin C. The first amino acid was not acetylated, but this difference from the muscle protein has no apparent effect on the function. The presence of Glu at position 99, as in turkey, versus Ala in chicken resulted in no detectable difference in comparing recombinant with chicken troponin C. A mutant in which residues 91-93 (Lys-Gly-Lys) in the D/E helical linker were deleted differs in function from wild-type troponin C in the conformational change that takes place upon calcium binding and its interaction with troponin I. Also, the mutant troponin C is impaired in its ability to form a functional complex with troponin I and troponin T that will confer calcium sensitivity on the actomyosin MgATPase. PMID- 2971048 TI - Fibromuscular dysplasia of renal arteries: comparison of intravenous digital subtraction angiography with conventional angiography. AB - Intravenous digital subtraction angiography (IV-DSA) was compared with conventional arteriography (CA) in 14 patients with fibromuscular dysplasia (FMD) of at least one renal artery. IV-DSA identified 29 of the 34 renal arteries detected by CA. A diagnostic quality IV-DSA examination was obtained in 23 of 29 renal arteries (78%). In adequately imaged renal arteries, IV-DSA correctly identified 12 of 20 FMD renal arteries, misdiagnosed 8 FMD renal arteries as normal and correctly identified 3 normal renal arteries. These poor results, due to poor spatial resolution and subtraction artifacts inherent in the IV-DSA system, warrant careful interpretation of negative examinations and further evaluation of high risk patients. PMID- 2971049 TI - Transcatheter embolotherapy of massive bleeding after surgery for benign gynecologic disorders. AB - Transcatheter embolotherapy is a well-established technique for control of bleeding in patients with pelvic trauma and pelvic malignancies, but it has been rarely used in massive vaginal bleeding after surgery for benign gynecologic disorders. We report five patients with massive post-operative vaginal hemorrhage following total abdominal hysterectomy (n = 3), repeated dilatation and curettage (n = 1) and cesarean section (n = 1), all successfully controlled with transcatheter embolization after many operative procedures had failed to stop the hemorrhage. The age of the patients ranged from 21 to 36 years. All patients underwent embolization of both internal iliac arteries with Gelfoam alone or in combination with stainless steel coils. The bleeding stopped immediately in all patients. Four patients recovered completely and one patient died of disseminated intravascular coagulation and multi-organ failure. PMID- 2971050 TI - Superselective cerebral arterial infusion of BCNU in high-grade glioma: the radiologist's point of view. AB - Thirty-two patients suffering from high-grade glioma were candidates for superselective cerebral arterial infusion of 1,3-bis-(2-chloroethyl)-1 nitrosourea (BCNU) after surgery and radiation therapy. There were 74 catheterizations using an 8-French guiding catheter through which a 2.5-French balloon catheter was placed into the main arterial trunk feeding the tumor. Eleven procedures were abandoned because of arterial spasm with a transient neurological deficit or because of prolonged catheterization time. Sixty-three infusions of BCNU were done, each lasting three hours. Eighty-one percent of patients showed stabilization or improvement on computed tomographic (CT) scans five weeks after treatment. We thus demonstrate the safety of supraophthalmic catheterization, the feasibility of prolonged catheterization, and the relative effectiveness of low doses of BCNU infused over a long period of time in the treatment of cerebral gliomas. PMID- 2971051 TI - Prosthetic heart valve malfunction: plain film findings. AB - Clinical signs of heart valve malfunction are often not specific so that recognition frequently depends on nonclinical methods. The chest radiographs of 34 patients with 41 prosthetic valve malfunctions (PVM) were compared before and after valve failure. The most frequent sign of PVM is postcapillary hypertension (90%). A marked increase in heart size occurs with regurgitation but not with obstruction. A change in attitude of a valve of more than 6 degrees. in the aortic and 12 degrees in the mitral position is virtually diagnostic of dehiscence. This was seen in 52% of patients with paravalvular regurgitation and in 29% of all patients with PVM. When dehiscence is suspected on clinical or radiologic grounds the valve should be examined fluoroscopically. Changes in the azygos vein and the vascular pedicle of the heart and the development of pleural effusions are less useful signs of PVM. Although the chest radiograph is often not diagnostic of PVM, it may point to the need for definitive investigation. PMID- 2971052 TI - Radiographic appearances of the St. Jude Medical Valve. AB - The St. Jude Medical Valve (SJMV), one of the more popular prosthetic cardiac valves in use today, has been described as poorly visualized on plain radiographs, being visible only at fluoroscopy. The chest radiographs of 76 patients (85 valves) in whom the SJMV had been implanted were reviewed in order to assess the visibility and appearances of the SJMV. We found that the SJMV was visible postoperatively in 70 of the 76 patients (92%) on either frontal or lateral chest radiographs or both. Knowing the various normal appearances of the SJMV described here and the anatomic locations of the heart valves, radiologists should be able to visualize and identify this prosthetic valve on chest radiographs in the majority of patients. PMID- 2971053 TI - CT appearance of the ventral pancreas. AB - Pancreas divisum has not been previously described in the computed tomography (CT) literature of the pancreas. Nine endoscopic retrograde choledochopancreatography (ERCP) confirmed examples of pancrease divisum were evaluated by CT. Four patients (44.4%) were observed to have a characteristic lobulated appearance of the pancreatic head. Of 21 patients with a normal pancreas confirmed by ERCP, only 2 (9.5%) showed lobulation on CT. The incidence of this previously unreported configuration was found to be significantly higher (p less than 0.05) in patients with pancreas divisum than in those with normal ductal anatomy. PMID- 2971054 TI - Real-time sonography of mediastinal and juxtamediastinal masses in infants and children. AB - Real-time sonography was used to evaluate 10 patients (age range, 2 days to 10 years) with mediastinal and juxtamediastinal masses. Seven of these also had computed tomography (CT). In eight patients sonography provided sufficient information with regard to probable diagnosis and adequate clinical management. CT was more helpful in two older children with postoperative seromas. Sonography is recommended as the imaging method of choice in neonates and in infants whose chest radiographs show mediastinal or juxtamediastinal masses. CT should be reserved for complex cases, and is of more value in older children. PMID- 2971055 TI - [Doughnut lesions of the cranial vault: an hereditary bone dysplasia]. AB - We describe a large family of whom twelve members were shown to have a benign bone dysplasia known as "doughnut lesions of the skull". Its clinical features are pathological fractures and cranial lumps; its radiological features comprise doughnut cranial lesions, double cortical lines of the vertebral bodies ("bone in bone"), squaring of the metatarsal and metacarpal bones, osteopenia, and tubulation defects of the diaphyses of the long bones. We believe the disease to be transmitted as an autosomal dominant trait. PMID- 2971056 TI - [Stress fractures of the femoral neck]. AB - Four young patients who developed a fatigue fracture of the femoral neck are presented. The clinical features of pain with movement are described. The role of nuclear medicine is discussed with emphasis on its importance in the early diagnosis of this entity. The radiologic stages based on the site and time of presentation are analyzed: 1) normal, 2) sclerotic bands perpendicular to the trabeculations, 3) incomplete fracture, 4) complete fracture. PMID- 2971058 TI - Cope drainage catheter: adding side holes without cutting the retention thread. AB - The retention thread of the Cope loop drainage catheter may be accidentally broken when additional side holes are cut into the catheter. We describe a simple technique of adding side holes which avoids this frustrating and expensive problem. PMID- 2971057 TI - The 30 degree cephalad anteroposterior tilt view to stage avascular necrosis of the femoral head. AB - A 30 degree cephalad anteroposterior tilt view of the femoral head was used, in addition to standard radiographic projections, in 55 patients having, or suspected of having, avascular necrosis of the femoral head. This method was then compared with standard views. Five patients with questionable findings on standard projections showed definite avascular necrosis, and 10 thought on standard views to have grade 2 disease were found to be grade 3, all from use of the additional projection. This projection should form part of the evaluation of suspected avascular necrosis of the femoral head. PMID- 2971059 TI - Dedifferentiated chondrosarcoma of bone with leiomyosarcomatous mesenchymal component: a case report. AB - Dedifferentiated chondrosarcomas are well-described aggressive tumors arising from low-grade chondrosarcomas and containing high-grade mesenchymal sarcomatous components. These tumors are important to consider in the differential diagnosis of well-defined lytic lesions in the pelvis associated with large soft-tissue components. We present the plain film, computed tomographic, and angiographic findings of the first such reported tumor containing a leiomyosarcomatous component. PMID- 2971060 TI - Longitudinal stress fracture of the tibia: case report. AB - Stress fractures usually present with vague clinical features and delayed radiographic findings. A patient with a longitudinal stress fracture of the tibia is described here. Computed tomographic images, perpendicular to the fracture, were diagnostic while plain radiographs and scintigraphy showed nondiagnostic changes due to an unexpected fracture pattern. PMID- 2971061 TI - Iliac artery aneurysms arising in patients having had resections of abdominal aneurysms. AB - We report the angiographic, computed tomographic, and ultrasonographic findings in two patients with iliac artery aneurysms detected several years after undergoing vascular graft surgery to the aorta. These lesions are often silent until serious complications occur, and they should be treated even when asymptomatic. Radiologists should be familiar with this complication so that it may be more readily diagnosed. PMID- 2971062 TI - Combined antegrade-retrograde catheterization of the occluded common iliac artery prior to angioplasty. AB - Catheterization of the totally occluded iliac artery prior to percutaneous transluminal angioplasty is frequently difficult. We report a patient in whom such an occlusion was traversed by a combined antegrade-retrograde approach, following which angioplasty was successfully performed. PMID- 2971063 TI - Intracranial subdural hematoma complicating metrizamide myelography. PMID- 2971064 TI - Torsion of a wandering spleen. AB - Splenic torsion is an uncommon cause of abdominal pain. It often leads to death and seldom has been diagnosed preoperatively. We report a patient in whom the anatomy of the lesion is illustrated, together with its predisposing factors and diagnostic features. PMID- 2971066 TI - Chylothorax associated with tuberculous spondylitis. AB - A patient is described with bilateral chylothorax which was associated with tuberculous spondylitis. The tuberculous spondylitis appeared at two widely separated thoracic levels. A similar association has not been reported previously. Anti-tuberculous treatment resulted in healing of the spondylitis and resolution of the chylothorax. PMID- 2971065 TI - Pyocolon: an unusual manifestation of colon ischemia. AB - An unusual manifestation of a large bowel ischemia was observed in two patients. Both had a left flank tubular fluid collection demonstrated respectively by computed tomography (CT) and ultrasonography (US). At surgery, pyocolon (i.e. ischemic, dilated large bowel segments filled with pus) was discovered and resected. Pyocolon should be suspected in patients with ischemic colitis and sepsis. US, although useful, may be misleading; CT is the investigation of choice to establish the diagnosis. PMID- 2971067 TI - The effect of body weight compression on axillo-femoral by-pass patency. AB - The influence of external compression on extra-anatomic bypass patency is still debated. The specific purpose of this study is the evaluation of ankle Pressure Index (P.I.) and Pulse Volume Recorder (P.V.R.) wave amplitude changes after 5 and 10 minutes of external bypass compression by body weight, lying on the side of the reconstruction. Eight patients with axillo-femoral bypass (mean follow-up 15.5 months) have been evaluated. The external body weight compression caused important changes of graft haemodynamics: (1) decrease in ankle Pressure Index at 5 min (p less than 0.005) or 10 min (p less than 0.0005); (2) decrease in P.V.R. wave amplitude at 5 min (p less than 0.005) and 10 min (p less than 0.025). PMID- 2971068 TI - Endothelial cell-derived heparan sulfate binds basic fibroblast growth factor and protects it from proteolytic degradation. AB - Cultured bovine capillary endothelial (BCE) cells were found to synthesize and secrete high molecular mass heparan sulfate proteoglycans and glycosaminoglycans, which bound basic fibroblast growth factor (bFGF). The secreted heparan sulfate molecules were purified by DEAE cellulose chromatography, followed by Sepharose 4B chromatography and affinity chromatography on immobilized bFGF. Most of the heparinase-sensitive sulfated molecules secreted into the medium by BCE cells bound to immobilized bFGF at low salt concentrations. However, elution from bFGF with increasing salt concentrations demonstrated varying affinities for bFGF among the secreted heparan sulfate molecules, with part of the heparan sulfate requiring NaCl concentrations between 1.0 and 1.5 M for elution. Cell extracts prepared from BCE cells also contained a bFGF-binding heparan sulfate proteoglycan, which could be released from the intact cells by a short proteinase treatment. The purified bFGF-binding heparan sulfate competed with 125I-bFGF for binding to low-affinity binding sites but not to high-affinity sites on the cells. Heparan sulfate did not interfere with bFGF stimulation of plasminogen activator activity in BCE cells in agreement with its lack of effect on binding of 125I-bFGF to high-affinity sites. Soluble bFGF was readily degraded by plasmin, whereas bFGF bound to heparan sulfate was protected from proteolytic degradation. Treatment of the heparan sulfate with heparinase before addition of plasmin abolished the protection and resulted in degradation of bFGF by the added proteinase. The results suggest that heparan sulfate released either directly by cells or through proteolytic degradation of their extracellular milieu may act as carrier for bFGF and facilitate the diffusion of locally produced growth factor by competing with its binding to surrounding matrix structures. Simultaneously, the secreted heparan sulfate glycosaminoglycans protect the growth factor from proteolytic degradation by extracellular proteinases, which are abundant at sites of neovascularization or cell invasion. PMID- 2971069 TI - Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C). AB - We recently found that the brain cytosolic microtubule-associated protein 1C (MAP 1C) is a microtubule-activated ATPase, capable of translocating microtubules in vitro in the direction corresponding to retrograde transport. (Paschal, B. M., H. S. Shpetner, and R. B. Vallee. 1987b. J. Cell Biol. 105:1273-1282; Paschal, B. M., and R. B. Vallee. 1987. Nature [Lond.]. 330:181-183.). Biochemical analysis of this protein (op. cit.) as well as scanning transmission electron microscopy revealed that MAP 1C is a brain cytoplasmic form of the ciliary and flagellar ATPase dynein (Vallee, R. B., J. S. Wall, B. M. Paschal, and H. S. Shpetner. 1988. Nature [Lond.]. 332:561-563). We have now characterized the ATPase activity of the brain enzyme in detail. We found that microtubule activation required polymeric tubulin and saturated with increasing tubulin concentration. The maximum activity at saturating tubulin (Vmax) varied from 186 to 239 nmol/min per mg. At low ionic strength, the Km for microtubules was 0.16 mg/ml tubulin, substantially lower than that previously reported for axonemal dynein. The microtubule-stimulated activity was extremely sensitive to changes in ionic strength and sulfhydryl oxidation state, both of which primarily affected the microtubule concentrations required for half-maximal activation. In a number of respects the brain dynein was enzymatically similar to both axonemal and egg dyneins. Thus, the ATPase required divalent cations, calcium stimulating activity less effectively than magnesium. The MgATPase was inhibited by metavandate (Ki = 5-10 microM for the microtubule-stimulated activity), 1 mM NEM, and 1 mM EHNA. In contrast to other dyneins, the brain enzyme hydrolyzed CTP, TTP, and GTP at higher rates than ATP. Thus, the enzymological properties of the brain cytoplasmic dynein are clearly related to those of other dyneins, though the brain enzyme is unique in its substrate specificity and in its high sensitivity to stimulation by microtubules. PMID- 2971070 TI - Cross-linking of IgE-receptor complexes by rigid bivalent antigens greater than 200 A in length triggers cellular degranulation. AB - We have examined the effect of cross-linking IgE-receptor complexes with variable receptor-receptor distances on the transmembrane signaling that leads to degranulation of rat basophilic leukemia cells. Linear polymers of the biotin binding protein avidin were generated with bis biotin-1,12-diamidododecane, and a dinitrophenyl-biotin conjugate was bound at each end of the polymers to form a series of rigid bivalent haptens of well-defined length. The polymers were fractionated by size with nondenaturing PAGE, electro-eluted, and tested for their ability to stimulate degranulation of rat basophilic leukemia cells sensitized with anti-DNP IgE. We found that hexamers of avidin (of length greater than or equal to 240 A) were as effective in triggering degranulation as dimers (of length approximately 80 A), while the monomeric avidin antigen (of length approximately 40 A) elicited a poorer degranulation response from the cells. The mechanism by which aggregation of cell surface receptors can initiate signal transduction is discussed in light of these results. PMID- 2971071 TI - Pancreas transplantation: international registry data. AB - The International Pancreas Transplant Registry received information on 1,157 pancreas transplants between December 1966 and April 1987. The results have progressively improved (p less than 0.001); graft and recipient survival rates at 1 year were 5% and 40% for 1966-1977 cases (N = 60), 20% and 71% for 1978-1982 cases (N = 205), and 43% and 80% for 1983-1987 cases (N = 92). In the 1983-1987 era, the graft survival rates have been similar (p less than 0.1) for the three most common duct management techniques, 46% at 1 year for polymer injection (N = 281), 44% for intestinal drainage (N = 253), and 44% for bladder drainage (N = 297), as well as for whole (N = 364) versus segmental (N = 528) grafts (41% versus 45% at 1 yr) and whether the spleen was (N = 27) or was not (N = 865) included (32% versus 44% at 1 yr). A preservation time of less than 6 hours (N = 557) was associated with higher graft functional survival rates than preservation times of 6 to 12 hours (N= 173) and greater than 12 hours (N = 67), 47%, 39%, and 35%, respectively at 1 year, but only the less than 6 versus the 6 to 12 hour difference was significant (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971072 TI - Type I diabetes as a chronic autoimmune disease. AB - The realization that Type I diabetes is an autoimmune disease and, in particular, a chronic autoimmune disease is beginning to impact on clinical care and research directed at elucidating the cause and prevention of diabetes. For example, specialized laboratory evaluation can now be used to exclude potential renal donors who are at high risk of developing diabetes (by screening renal donor candidates who are relatives of Type 1 diabetics for cytoplasmic islet cell antibodies and evaluating first phase insulin secretion on intravenous glucose tolerance testing). The most important long-term consequence of the ability to predict Type 1 diabetes may be the development of effective immunotherapy to prevent the disease. Finally, the realization that Type 1 diabetes is an auto immune disease and that some of the antigens expressed by islets (e.g., specific gangliosides identified with monoclonal antibodies) are expressed by renal glomerular cells, retinal microvascular pericytes, and neurons has renewed interest in searching for immunologic factors contributing to secondary complications. PMID- 2971073 TI - Role of altered capillary hemodynamics in the initiation and progression of diabetic microangiopathy. PMID- 2971074 TI - Vitrectomy for complications of proliferative diabetic retinopathy. PMID- 2971075 TI - Acid-base disorders in hyperglycemia of insulin-dependent diabetic patients on chronic dialysis. AB - The authors studied hyperglycemia occurring in insulin-dependent diabetic patients on chronic dialysis to determine the types of associated acid-base disorders, their treatment, and any differences from hyperglycemia in diabetic patients with intact renal function. Eighty-eight episodes of serum glucose greater than 25 mmol/L were observed, 23 in hemodialysis patients and 65 in patients on continuous peritoneal dialysis. Treatment consisted of low-dose insulin in 77 episodes and low-dose insulin plus saline in 11; no base was administered. Seventeen episodes (19%) presented with ketoacidosis. Arterial blood gas determinations were carried out at presentation in 37 of the episodes without ketoacidosis. Of these, 12 had respiratory alkalosis, six had respiratory acidosis and severe pulmonary edema, 14 had other single or mixed acid-base disorders, and only five had normal acid-base status. Insulin corrected the ketoacidosis in all instances and both pulmonary edema and respiratory acidosis in five of six instances. In eight cases metabolic alkalosis developed during treatment, without external acid loss. At the completion of treatment respiratory alkalosis was present in half the cases. No difference was noted between patients treated with hemodialysis or peritoneal dialysis. Insulin alone is sufficient for the management of hyperglycemia in dialysis patients. Certain acid-base disorders persist, but do not need further treatment. Hyperglycemia in patients on dialysis is characterized by infrequent development of metabolic acidosis and frequent presentation with respiratory alkalosis, by respiratory acidosis that is corrected by insulin, and by metabolic alkalosis developing during treatment without external cause. PMID- 2971076 TI - The Wisconsin Epidemiologic Study of Diabetic Retinopathy: VIII. The incidence of retinal photocoagulation. AB - The incidence of focal and panretinal photocoagulation and its relationship to demographic and other characteristics were examined in a population-based study of people with diabetes in southern Wisconsin. For participants whose age at diagnosis was less than 30 years, who were taking insulin, and who had not been previously treated with photocoagulation, the 4 year incidence of panretinal photocoagulation (10.8%) was significantly higher (p less than .0001) than the rate of focal and/or grid photocoagulation of the macula (4.3%). For those whose age at diagnosis was 30 years or older and who had not been previously treated with photocoagulation, the incidence rates of panretinal photocoagulation (4.4%) and focal and/or grid photocoagulation of the macula (3.1%) were not significantly different (p = .11). At follow-up examination, 33.8% of the eyes of younger onset persons and 57.7% of the eyes of older onset persons with Diabetic Retinopathy Study high risk characteristics for severe visual loss had never received panretinal photocoagulation. These relatively high frequencies of untreated eyes in need of panretinal photocoagulation remain a concern. PMID- 2971077 TI - Reduced urinary kallikrein in hypertensive diabetic patients. PMID- 2971078 TI - High blood pressure is a major factor in progression of diabetic nephropathy. AB - Arterial hypertension is a frequent finding, even early in the course of diabetic nephropathy. Systemic and glomerular hypertension enhance the development of diabetic glomerulopathy and accelerate the rate of decline in glomerular filtration rate in diabetic nephropathy. Conversely, effective antihypertensive treatment reduces albuminuria and diminishes the rate of decline in glomerular filtration rate, thereby postponing end-stage renal failure in diabetic nephropathy. PMID- 2971079 TI - The insulin-like growth factor II/mannose-6-phosphate receptor is present in monkey serum. AB - We recently reported that the insulin-like growth factor II (IGF-II)/mannose-6 phosphate (Man-6-P) receptor is present in fetal and postnatal rat serum and that its serum content declined dramatically postnatally between days 20 and 40 . We now provide evidence that the IGF-II/Man-6-P receptor is also present in monkey serum. Serum was gel filtered on Sephadex G-200, and the column fractions were assayed for binding of radiolabeled IGF-II. There was significant binding of [125I]IGF-II to the void volume fractions in addition to binding to the 150K and 40K carrier proteins. Binding to the void volume fractions was greatest in cord serum and decreased with age. Competitive binding studies with [125I]IGF-II and the void volume pools from monkey serum demonstrated that IGF-I competed less potently than IGF-II, and insulin did not compete. Radiolabeled IGF-I did not bind specifically to the void volume pools. Chemical cross-linking of [125I]IGF II to aliquots of the void volume pools from monkey cord serum samples and analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence of dithiothreitol demonstrated a specific band at about 240K. Western blotting using a specific antiserum (no. 3637) against rat IGF-II/Man-6-P receptor was performed on aliquots of the Sephadex G-200 void volume pools of monkey serum. A band of approximately the same size as that found with human fibroblast members (approximately 215 K without dithiothreitol) was detected. The IGF-II/Man-6-P receptor band was more intense in cord serum than in the postnatal samples. When cord serum Sephadex G-200 pools were gel filtered on Sephadex G-50 in 1 mol/L acetic acid to separate binding components from free IGF, and IGF-II was measured by RRA, approximately 20% of the circulating IGF-II was found to be associated with this IGF-II/Man-6-P receptor in monkey serum. We conclude that the IGF-II/Man-6-P receptor present in serum may be a significant carrier for IGF II in the monkey. PMID- 2971080 TI - The effect of a gonadotropin-releasing hormone agonist analog (nafarelin) on bone metabolism. AB - The effect on bone metabolism of an agonist analog of GnRH, nafarelin, was studied in 16 premenopausal women, who received 200 micrograms nafarelin/day for 6 months, and 9 premenopausal women, who received 400 micrograms nafarelin/day for 6 months, followed by a 6-month follow-up period. Bone mineral content in the forearm (measured by single photon absorptiometry) and in the spine (measured by dual photon absorptiometry) significantly decreased after 6 months of treatment with 400 micrograms nafarelin, but 6 months after termination of treatment all bone mineral measurements had returned to pretreatment levels. The bone mineral measurements in the 200 micrograms group did not change throughout the study. In both treatment groups the biochemical estimates of bone turnover increased significantly to postmenopausal levels. Withdrawal of treatment resulted in an abrupt decrease in the bone resorption parameters (fasting urinary hydroxyproline to creatinine and calcium to creatinine excretion ratios and serum phosphate), whereas there was a protracted fall in the bone formation parameters (plasma bone Gla protein and serum alkaline phosphatase) 6 months after termination of treatment. Our findings demonstrate that nafarelin in both doses increased biochemical indices of bone turnover, that 400 micrograms/day nafarelin resulted in a significant decrease in bone mineral content, and that these effects were reversible. PMID- 2971081 TI - In vitro induction of anti-thyroid microsomal antibody-secreting cells in peripheral blood mononuclear cells from normal subjects. AB - Secretion of immunoglobulin G (IgG) and IgM antithyroid microsomal antibodies (AMA) was induced in vitro by coculturing non-T cells (B lymphocytes) and autologous CD4 (helper/inducer) cells from normal subjects stimulated with pokeweed mitogen (PWM) or a combination of human thyroid microsomal antigen (McAg) and Staphylococcus aureus Cowan I (SAC) strain. With PWM stimulation, AMA production was induced in more IgM-secreting cells (AMA-M) than IgG-secreting cells (AMA-G). However, McAg plus SAC stimulation resulted in similar numbers of AMA-G- and AMA-M-secreting cells. PWM induced a significantly greater number of both AMA-M (and generalized IgM)-secreting cells than did McAg plus SAC, while the number of AMA-G-secreting cells induced by the two stimuli were similar. There were no significant differences between autologous or allogeneic CD4 cells from normal subjects or patients with autoimmune thyroid disease (AITD) when cocultured with B cells from normal subjects in terms of helper activity in the induction of AMA-M- or IgM-secreting cells with PWM stimulation. However, with McAg plus SAC, CD4 cells from patients with AITD induced a significantly greater number of AMA-M-secreting cells than did autologous or allogeneic CD4 cells from normal subjects. There was no difference in helper activity between autologous and allogeneic normal CD4 cells in the induction of generalized IgM-secreting cells regardless of the stimulus used. Normal autologous or allogeneic CD8 (suppressor/cytotoxic) cells cocultured with normal B cells and autologous CD4 cells suppressed the induction of AMA-M-secreting cells by PWM stimulation. On the other hand, CD8 cells from patients with AITD suppressed the induction of AMA M-secreting cells significantly less effectively. All CD8 cells suppressed the induction of IgM-secreting cells equally well. We conclude that 1) B lymphocytes from normal subjects are capable of producing autoantibodies in vitro in the presence of CD4 cells; 2) the helper activity of CD4 cells from patients with AITD to induce AMA-M secreting cells is greater than that of normal CD4 cells with thyroid antigen stimulation; and 3) this helper activity may be due to relatively impaired suppressor activity in thyroid antigen-specific CD8 cells from patients with AITD, whereas the immunoregulatory function of CD8 cells from normal subjects appears to play an important role in the maintenance of self tolerance. PMID- 2971082 TI - Ovarian hyperstimulation for in-vitro fertilization controlled by GnRH agonist administered in combination with human menopausal gonadotrophins. AB - Surges of luteinizing hormone (LH) in serum that result in luteinization, but occur prematurely with respect to the diameter of the leading follicle, frustrate attempts to induce multiple follicular maturation for in-vitro fertilization (IVF) in a number of women. We examined the possibility of blocking premature LH surges by the administration of D-TRP6-LH-RH, a potent agonistic analogue of gonadotrophin-releasing hormone (GnRH). Six patients who had repeatedly shown premature LH surges were treated for 10 days, beginning between days 1 and 3 of the cycle with daily s.c. injections of 500 micrograms D-Trp6-LH-RH followed by a daily injection of 100 micrograms of the analogue until the day of administration of human chorionic gonadotrophin (HCG). When pituitary and ovarian suppression had occurred, ovarian stimulation with human menopausal gonadotrophin was started and adjusted in dose according to the ovarian response. HCG was injected when the dominant follicle had reached a diameter of at least 18 mm and oestradiol levels were above 300 pg for each follicle greater than 15 mm. Oocyte collection was performed 36 h later via laparoscopy, followed by IVF and embryo transfer. The six patients studied to date responded to therapy and treatment could be completed up to embryo transfer. Two patients became pregnant; one of the pregnancies, however, resulted in abortion. Combined treatment with GnRH analogue for suppression of pituitary gonadotrophin secretion followed by the administration of gonadotrophins thus seems to be a promising method for ovarian stimulation in patients who frequently exhibit premature LH discharges and therefore fail to complete treatment. PMID- 2971083 TI - Hypotensive effect of taurine. Possible involvement of the sympathetic nervous system and endogenous opiates. AB - We studied the role of diminished sympathetic nervous system (SNS) activity and endogenous opiate activation in the hypotensive action of taurine, a sulfur amino acid, in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Supplementation of taurine could prevent the development of DOCA-salt hypertension in rats, but failed to change blood pressure in vehicle-treated control rats. Cardiac NE turnover, which was determined from the rate of decline of tissue NE concentration after the administration of alpha-methyl-p-tyrosine, was markedly accelerated in DOCA-salt rats, but 1% taurine supplement restored it to normal. Moreover, naloxone (2 mg/kg), the specific opiate antagonist, increased blood pressure in taurine-treated DOCA-salt rats, restoring it to levels similar to those in the DOCA-salt rats. In contrast, taurine did not decrease cardiac NE turnover in the control rats, nor did naloxone increase blood pressure in the taurine-treated control rats. Moreover, supplementation of taurine increased both beta-endorphin-like immunoreactive material and taurine contents in the hypothalamus of DOCA-salt rats, whereas it did not increase beta endorphin in that of control rats despite increased taurine contents. Thus, taurine not only normalized the increased cardiac SNS activity but also elicited an opiate-mediated vasodepressor response only in DOCA-salt rats. It is suggested, therefore, that endogenous opiate activation, which is intimately related to SNS suppression, may contribute to the antihypertensive effect of taurine in sodium chloride hypertension. PMID- 2971084 TI - Identification of high risk relatives for coronary heart disease. AB - The family at increased risk for future coronary heart disease is the family with a member who has 1) had one or more myocardial infarctions before age 55 years; 2) has levels of LDL cholesterol greater than 75th percentile for age; 3) has excessively low levels of HDL2 cholesterol; 4) has hypertension or has had a stroke, or both; 5) has excessive weight at any age and excessive weight gain during adulthood, or 6) smokes in the household. PMID- 2971085 TI - Doppler analysis of flow velocity profile at the aortic root. AB - This study of aortic root flow velocity profiles suggests that the flow velocity pattern is parabolic rather than flat as has been widely supposed. To analyze the distribution of flow velocity, a special ultrasound pulsed Doppler velocity flow meter, with a direct probe on the aortic wall, was designed. Sixteen patients, none with a diagnosis of aortic valve disease, were examined just before undergoing open heart surgery. Parabolic velocity patterns were always found, and it was determined that the parabola was skewed rightward toward the anterior wall. Furthermore, it was shown that the flow velocity vector was initially directed toward the commissure of the right coronary and the noncoronary cusps. PMID- 2971087 TI - Systolic and diastolic flow abnormalities in elderly patients with hypertensive hypertrophic cardiomyopathy. AB - Seventeen patients with clinical and echocardiographic features of hypertensive hypertrophic cardiomyopathy of the elderly were studied to more completely characterize left ventricular systolic and diastolic function in this group. Measurements of left ventricular structure and systolic and diastolic function were made in the study patients and compared with those of age-matched control subjects. The study group had significantly greater left ventricular mass, wall thickness, shortening fraction and relative wall thickness than did the control subjects. Left ventricular end-diastolic dimension was smaller and left atrial size was not different in study patients compared with control subjects. Left ventricular filling was characterized by an increased peak atrial velocity and reduced ratio of peak early to peak atrial velocity in the study group. Left ventricular outflow velocities were elevated in 14 of the 17 study patients with peak velocities ranging from 1.2 to 5.0 m/s corresponding to a peak intraventricular gradient of 16 to 100 mm Hg. The velocity waveforms in these patients were late-peaking, similar to those described in hypertrophic obstructive cardiomyopathy. The elevated velocities were localized to the left ventricular outflow tract. These findings imply a pathophysiologic state in these elderly patients with long-standing hypertension, very similar to that in hypertrophic obstructive cardiomyopathy, and provide further support for the use of pharmacologic agents with negative inotropic properties or positive lusitropic properties in this group. PMID- 2971086 TI - Incomplete redistribution in delayed thallium-201 single photon emission computed tomographic (SPECT) images: an overestimation of myocardial scarring. AB - To determine if incomplete redistribution at 4 h in exercise tomographic thallium 201 studies is always due to a myocardial scar, 141 patients were evaluated before and after a total of 160 successful percutaneous transluminal coronary angioplasty procedures. Thallium studies were analyzed using polar "bull's-eye" maps. For both immediate and delayed images, abnormalities were quantified as a thallium score by calculating a standard deviation-weighted sum of pixels greater than 2.5 SD below gender-matched normal limits. One hundred forty-four of 160 studies indicated abnormalities before angioplasty. Of these 144, incomplete redistribution occurred in 111 (77%): 16 (14%) in patients with and 95 (86%) in patients without prior Q wave myocardial infarction. After angioplasty, improvement in delayed image score occurred in 8 (50%) of 16 patients with prior infarction and 72 (76%) of 95 patients without prior infarction (p less than 0.05). After angioplasty, delayed images were normal in 1 (6%) of 16 patients with prior infarction and 32 (34%) of the 95 without (p less than 0.05). Before angioplasty, delayed image scores were positively correlated with scores in the immediate postexercise images in patients with (r = 0.84) and those without (r = 0.69) prior infarction. To determine if additional delayed images could help differentiate scar from ischemia, an 8 to 24 h delayed image was obtained in each of 40 other patients with incomplete redistribution at 4 h. Of 28 patients without prior infarction, 15 had no redistribution, and 13 had further redistribution at 8 to 24 h. PMID- 2971088 TI - A longitudinal study of functional change and mortality in the United States. AB - Studies of functional impairments in the U.S. elderly population have tended to rely on prevalence estimates from nationally representative health and institutional surveys. These prevalence estimates generally show higher rates of disability for females than males. Unfortunately, prevalence estimates can be misleading when one attempts to assess the risks of certain types of health event transitions for individuals. This study directly examined the individual transitions both into and out of functionally impaired states using longitudinal data from the 1982 and 1984 National Long Term Care Surveys (NLTCS). The data show that, even at very high levels of impairment, there are significant numbers of community residents who apparently manifest long-term improvement in functioning. The longitudinal data also show that the risks of becoming disabled are roughly the same for males and females. This suggests that sex differences in the national prevalence of disabilities arise from the greater longevity of females at any given level of age and functional impairment. PMID- 2971089 TI - Medical need and use of services among older men and women. AB - This article presents a model of the process by which men and women evaluate their health and utilize the medical care system. It is argued that an analysis of the effects of sex roles on medical care must look at the entire process and not just focus on the outcome variables of physician visits and hospitalization. Gender interactions are emphasized, but race interactions also are examined. Findings show that men and women differ in the number of illnesses they report, the likelihood of having a life-threatening illness, the degree of disability, and their perception of their health. Indicators of medical need were found to be the strongest measures in predicting the recency of seeing a physician; however, the physician visit had the strongest effect in determining hospitalization. No direct effects of gender or race on physician visits were found, and only one interaction was significant; however, the results were very different when the measure of medical care use was hospitalization. Both additive and interactive effects by gender and race were found when predicting hospitalization. PMID- 2971090 TI - Penetrating duodenal ulcer from hepatic artery chemotherapy infusion catheter. Successful conservative medical management. AB - A 45-year-old woman receiving continuous infusions of 5-fluoro-2'-deoxy-uridine into the hepatic artery through a surgically implanted intraarterial catheter for hepatic metastases from adenocarcinoma of the colon, developed acute epigastric pain and anemia. Endoscopy demonstrated a duodenal ulcer with a portion of the catheter visible in the crater. Conservative management with sucralfate and ranitidine allowed endoscopically proven healing of the penetrating ulcer with subsequent successful reuse of the catheter. PMID- 2971091 TI - Light and electron microscopic localization of atrial natriuretic peptide in the heart of spontaneously hypertensive rat. AB - Atrial natriuretic peptide (ANP) is a newly discovered peptide hormone present mainly in the atria. We investigated the occurrence and distribution of ANP immunoreactivity in the myocardiocytes of the ventricles of spontaneously hypertensive rats by use of immunocytochemistry at both light and electron microscopic level. ANP immunoreactivity was found in the specific granules in the cytoplasm of the cardiocytes in the subendocardium and the myocardium of the ventricles, as well as in the atria. The specific granules found in the ventricles of hypertensive rats were similar in size, shape, and ANP immunoreactive content to those in the atria. The abundance of ANP immunoreactivity in the left ventricle is greater than that in the right, and appears to increase with increasing severity of hypertension. Conversely, the overall content of ANP in the atria of hypertensive rats was decreased when compared with that in age-matched normotensive rats. The present findings indicate that ventricles may become a major source for ANP synthesis and release during hypertension, and may play important roles in cardiac endocrine pathology and cardiac hypertrophy. PMID- 2971092 TI - An immunocytochemical method for the detection of fluorochrome-labelled DNA probes hybridized in situ with cellular RNA. AB - Various detection systems for in situ hybridization of nucleic acids are currently used. We report here an immunocytochemical detection system which is based on the detection of FITC-labelled DNA/mRNA hybrids and takes advantage of FITC molecules attached covalently to the DNA probes prior to hybridization. In situ hybridization on cytocentrifuge spots is followed by the application of an anti-fluorescein antibody thus permitting detection of mRNA/DNA-FITC hybrids. The anti-FITC antibody reaction is demonstrated by an indirect immunocytochemical peroxidase-staining method. The T lymphoblast cell line Jurkat and the cDNA for the TCR-beta chain were chosen to establish the technique. PMID- 2971093 TI - [Study of the biological activities of toxic shock syndrome toxin-1 (TSST-1). III. Analysis of TSST-1-induced suppression of the primary antibody response]. PMID- 2971094 TI - [Endocrinological and electron microscopical studies on the complete form of testicular feminization syndrome]. AB - A 23-year-old patient with a complete form of testicular feminization syndrome (Tfm) was examined endocrinologically and morphologically by electron microscopy. 1. Serum testosterone, DHEA and estrone concentrations and the urine 17-KS concentration were higher than in adult males. Furthermore, serum pituitary gonadotropins, especially LH, were increased. 2. We found that testicular venous testosterone, estrone, estradiol and progesterone were remarkably increased compared with their peripheral concentrations, and that circulating concentrations decreased rapidly after gonadectomy. The electron microscopic study showed active steroidogenesis in Leydig cells. We believe that these results prove that these steroids are produced in Leydig cells. 3. We found that the administration of 10,000 IU/day of hCG for 2 days increased serum gonadal steroids but that of 225 IU/day of hMG for 2 days did not, that LH responded strongly to LH-RH but FSH response was weak, and that after gonadectomy serum LH immediately increased but FSH did not change for at least 24 hours. From these results, we assume that FSH is not related to steroidogenesis and that the mechanisms regulating serum LH and FSH are different. 4. The seminiferous tubules were occupied by two kinds of cells. Both kinds were Sertoli cells, one of which was numerous and of low electron density indicating an immature type, and the other was of high electron density indicating a mature type. Spermatogonia were not detected. PMID- 2971095 TI - Blood flow velocity in cutaneous lesions of leprosy. AB - The velocity of blood flow in the cutaneous lesions of leprosy was measured by the noninvasive technique of laser-Doppler velocimetry in nine male patients, all under treatment with World Health Organization multidrug regimens. In three patients with stable borderline tuberculoid (BT) lesions, the blood-flow velocity was slightly faster over the lesion than that in adjacent uninvolved skin. There was no substantial difference between different sites at the center and edge of individual lesions in each patient, but there was some variation between subjects. In one other BT patient with a reversal reaction, blood-flow velocity was 20-fold greater than in adjacent uninvolved skin. In four patients with stable borderline lepromatous/lepromatous (BL/LL) disease, the blood-flow velocity was 3 to 9 times faster over the plaques than in adjacent skin. There was relatively little difference between measurements over comparable points at the edge and center of individual plaques, or between plaques in the same patient, but there were considerable differences between patients. A fifth BL patient in reversal reaction showed generally similar results to those found in the stable BL/LL patients. Histometric study of the biopsies taken immediately after blood-flow measurement from two different plaques on each of four BL patients showed a clear relationship between the granuloma fraction measured by planimetry and the blood-flow velocity. This finding suggests that laser-Doppler velocimetry might prove to be a useful, clinically acceptable, noninvasive technique for monitoring the severity of hyperemia as an early indication of reversal reaction during chemotherapy trials in leprosy patients. PMID- 2971096 TI - Comedones induced by coconut oil in a borderline tuberculoid lesion. PMID- 2971097 TI - Pharmacological and biochemical characterization of rat hippocampal 5 hydroxytryptamine1A receptors solubilized by 3-[3 (cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS). AB - Rat hippocampal 5-hydroxytryptamine1A (5-HT1A) binding sites were solubilized with a yield of 34% using 3-[3-(cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS, 10 mM) as detergent. Kinetic analyses of [3H]8-hydroxy-2-(di-n propylamino)tetralin ([3H]8-OH-DPAT) binding indicated that the 5-HT1A sites exhibit the same properties in the soluble form as in the membrane-bound form. Furthermore, a positive correlation (r = 0.988) was found between the respective pIC50 values of a series of agonists and antagonists to inhibit [3H]8-OH-DPAT binding to either soluble or membrane-bound 5-HT1A sites. Gel filtration through Sephacryl S-400 as well as chromatography on wheat germ agglutinin (WGA)-agarose did not affect the modulation by guanine nucleotides (5' guanylylimidodiphosphate) of [3H]8-OH-DPAT binding which suggests that the 5-HT1A binding subunit is a glycoprotein tightly attached to a G protein even in its soluble form. The [3H]8-OH-DPAT binding material eluted from Sephacryl S-400 had an apparent molecular mass of 155 kilodaltons, as expected from a heterodimer with one binding subunit (approximately 60 kilodaltons) and one G protein (approximately 80 kilodaltons). Marked enrichment in 5-HT1A binding sites relative to other soluble proteins was found in the peak fractions eluted from Sephacryl S-400 (by sixfold) and WGA-agarose (by 26-fold) columns, suggesting that these chromatographic steps might be of interest for the purification of central 5-HT1A receptors. PMID- 2971098 TI - Differentiation of dopamine overflow and uptake processes in the extracellular fluid of the rat caudate nucleus with fast-scan in vivo voltammetry. AB - Stimulated overflow of dopamine (DA) into the extracellular fluid of the rat caudate nucleus was measured with fast-scan cyclic voltammetry. DA concentrations were sampled in less than 10 ms at 100-ms intervals with a Nafion-coated, carbon fiber microelectrode. Overflow of DA was induced by electrical stimulation of the medial forebrain bundle with 300-microA pulses of various duration and frequency. Stimulated overflow was measured as a function of stimulus duration before and after administration of benztropine, bupropion, and amphetamine. These results were correlated with simulated curves based on a simple uptake/overflow model. The observed overflow was assumed to be a function of [DA]p, the concentration of DA which overflows per stimulus pulse, and the kinetics of cellular uptake of DA. Correlation of experimental with stimulated results was obtained at the 95% confidence limit for the duration studies; however, it was not possible to distinguish between the effects of pharmacological agents on uptake and overflow. In contrast, modulation of stimulus frequency did permit such distinction. Simulations of an increase in [DA]p fit results following dihydroxyphenylalanine methyl ester at 95% confidence limits, whereas an equivalent change in the apparent Km did not fit. An increase in the apparent value of Km correlated with results obtained at different frequencies following nomifensine and bupropion administration at the 95% confidence limit, whereas an equivalent increase in [DA]p did not fit. The effects of GBR 12909 best correlated with an increase in the DA available for overflow. PMID- 2971099 TI - The Framingham Disability Study: physical disability among community-dwelling survivors of stroke. AB - The relationship between stroke and physical disability was examined in a cohort of adult, Framingham, Massachusetts, residents who, between 1948 and 1951, were assembled for a longitudinal examination of cardiovascular disease. Multivariate analyses examined the amount of residual disability attributable to stroke among 2540 community-dwelling survivors, 27 years after their initial examination, after controlling for age, cardiovascular risk factors, other cardiovascular diseases, and eight general health conditions related to physical disability. Among men living in the community, a history of stroke explained 12% of the variance in physical disability. Suffering a stroke, however, was not as strongly related to physical disability among women living in the community, accounting for only 3% of the variance. Results suggest that although older men and women die from the same major causes, they may not be disabled by the same conditions. PMID- 2971100 TI - Quantification of thermal asymmetry. Part 2: Application in low-back pain and sciatica. AB - Temperature differences between the lower extremities were measured using a computerized thermometric scanning system in order to compare the degree of thermal asymmetry in 144 patients with low-back pain. The patients displayed highly significant thermal asymmetries, with the involved limb being cooler (p less than 0.001). When asymmetries exceeded 1 standard deviation from the mean temperature of homologous regions measured in 90 normal control subjects, the positive predictive value of thermometry in detecting root impingement was 94.7% and the specificity was 87.5%. These values indicate that calculation of temperature asymmetry is particularly effective in evaluating reported pain in psychosocially affected patient populations in whom the chance of positive myelography or impaired root function is low. In this group of patients, thermometric study provides physicians with important information for proper decision making. The test can be performed to avoid more invasive and probably less revealing diagnostic or exploratory surgical procedures. PMID- 2971101 TI - Adoptive cell transfer studies to examine the role of T lymphocytes in immunity to Trypanosoma musculi. AB - Previous studies in this laboratory utilizing monoclonal antibody-induced immunosuppression have demonstrated that the T-helper lymphocyte is primarily responsible for the T lymphocyte dependency of Trypanosoma musculi elimination from the bloodstream of mice, and that T-cytotoxic lymphocytes play a minimal role in this response. In the current study, these findings were extended by examining the effects of adoptive cell transfers on the course of infection with T. musculi using immune splenocytes enriched for T lymphocyte subpopulations. These studies demonstrated that adoptive transfer of immune splenic T lymphocytes resulted in a specific, dose-related enhancement of kinetics of trypanosome elimination. This effect was found to be due to the presence of L3T4+ T-helper cells in the immune splenocyte population. Adoptive transfer of Lyt-2+ T cytotoxic cells or lymphokine-activated killer (LAK) cells was ineffective in altering the course of infection. In addition, it was found that immune B lymphocytes were equally capable of adoptively transferring immunity to T. musculi, suggesting that the primary role of the T-helper lymphocyte is to provide help in the induction of parasite-specific antibodies. PMID- 2971102 TI - Mononuclear cell subsets in patients with different clinical forms of paracoccidioidomycosis. AB - Seventy untreated paracoccidioidomycosis patients, 15 with the acute or subacute form of the disease and 55 with the chronic form, were compared with two normal control groups of the same age range. Peripheral blood mononuclear cell subsets were defined by monoclonal antibodies directed at total T cells, helper/inducer and suppressor/cytotoxic T cell subpopulations; B cells, cortical thymocytes and monocyte/null cells. Both groups of patients showed an increased number of monocyte/null cells, a low helper/suppressor ratio and a reduced percentage of total T cells and their helper/inducer subsets. In addition patients with the acute form of the disease exhibited high levels of suppressor/cytotoxic T cells and B cells. These findings are of importance in our attempts to understand the pathogenesis of this mycosis and also to evaluate its prognosis in individual patients. PMID- 2971103 TI - Severity of intestinal damage in gastroschisis: correlation with prenatal sonographic findings. AB - From 1982 to 1986, 26 abdominal wall defects were detected prenatally. Seventeen pregnancies culminated in live births with ultrasound correctly distinguishing between omphalocele and gastroschisis in all cases. In the 11 cases of gastroschisis diagnosed before birth, we attempted to correlate the clinical outcome with the size of the abdominal wall defect, sonographic appearance of the eviscerated bowel, and known time of exposure to amniotic fluid (gestational age at birth to gestational age at diagnosis). Neither the time of exposure to amniotic fluid (median duration of 14 weeks) nor the defect size could be correlated with eventual clinical outcome. The presence of small bowel dilatation and mural thickening on prenatal sonography (four patients) had a high correlation with severe intestinal damage and poor clinical outcome. The absence of these two sonographic findings (seven patients) was associated with mild intestinal changes and benign clinical course with no morbidity or mortality. We conclude that obstetric ultrasound cannot only accurately detect the presence and type of abdominal wall defect, but it also gives an indication of the severity of intestinal damage and subsequent clinical course in prenatally diagnosed cases of gastroschisis. Early delivery of the fetus with prenatally diagnosed gastroschisis should no longer be performed to limit exposure to amniotic fluid. Now that reliable sonographic criteria of severe intestinal damage have been defined, the decision to deliver early can be restricted to those fetuses with bowel dilatation and mural thickening. PMID- 2971104 TI - Hemodynamic and renal effects of atrial natriuretic factor in portal hypertensive rats. Potentiation by Phe-Ile-Orn-vasopressin. AB - The effects of atrial natriuretic factor (ANF) on splanchnic hemodynamics and renal function in portal hypertensive models are described incompletely. Furthermore, ANF-induced vasodilatation and hypotension may limit the assessment of its own renal physiological effects. We infused ANF (human ANF 102-126) to anesthetized portal vein-ligated rats, a model with prehepatic portal hypertension. Arterial pressure was reduced by 17%, but portal pressure was unaffected. Diuresis and natriuresis were explained in part by an increase in glomerular filtration rate; in addition, renal vascular resistance was significantly decreased. The natriuretic response to ANF was slightly, but significantly, decreased in portal hypertensive rats as compared to controls (fractional excretion of sodium, 1.8 +/- 0.4 vs. 2.9 +/- 0.3; P less than .05). The addition of Phe-Ile-Orn-vasopressin, a V1 receptor agonist, normalized arterial pressure but induced a significant decrease in portal pressure (15 +/- 0.9 mm Hg base line vs. 12.8 +/- 0.7 combination group; P less than .01). Furthermore, the combination of both drugs markedly potentiated the natriuretic effects (0.4 +/- 0.1 microEq/min of control vs. 10.0 +/- 2.3 ANF vs. 32.2 +/- 3.3 combination group; P less than .001). The natriuretic potentiation resulted from increments in glomerular filtration rate and renal blood flow. Normalization of arterial pressure may enhance the renal physiological effects of ANF, in this portal hypertensive model. PMID- 2971105 TI - Stretch sensitization of human muscle spindles. AB - 1. Sixty-seven afferents from the finger extensor muscles were consecutively recorded by microneurography. 2. The units were classified as primary or secondary muscle spindle afferents or Golgi tendon organ afferents on the basis of their responses to ramp-and-hold stretches, sinusoidals superimposed on ramp and-hold stretches, maximal twitch contractions and isometric contractions and relaxations. 3. The muscle was repeatedly stretched and then either kept short or long for a few seconds followed by a slow ramp stretch. The responses of the muscle afferents to the slow stretch were compared under the two conditions. 4. Thirty out of thirty-eight units classified as primary spindle afferents and four out of eleven units classified as secondary afferents showed an enhanced response to the slow ramp when the muscle had been kept short compared to the response when the muscle had been kept long. 5. None of the eighteen Golgi tendon organ afferents showed any difference in this respect. 6. It is concluded that stretch sensitization does occur in human muscle spindles and, when present, constitutes firm evidence of the afferent originating from a muscle spindle rather than a Golgi tendon organ. In addition, due to differences in the response characteristics of primaries and secondaries, the test may aid in separating muscle spindle primary afferents from secondary afferents. PMID- 2971106 TI - Outpatient laparoscopy. PMID- 2971107 TI - Nursing care of the patient undergoing laparoscopy in the ambulatory setting. PMID- 2971108 TI - Successful management of ruptured suprarenal aortic aneurysm with left artery stenosis (a case report). PMID- 2971109 TI - Surgery and balloon angioplasty in advanced renovascular hypertension(46 cases). PMID- 2971110 TI - The effect of human pregnancy serum on the synthesis and action of interleukin-1. AB - Supernatants from mixed lymphocyte reactions performed in the presence of pregnancy serum (PS) or normal human serum (NHS) were tested for interleukin-1 (IL-1) activity in a thymocyte assay. A lower response was obtained with PS containing supernatants, suggesting that PS exerts its suppressive effect by inhibiting IL-1 synthesis or by blocking its mode of action. IL-1 activity generated by human peripheral blood mononuclear cells (PBMC) stimulated with LPS and cultured in the presence of PS or NHS was similar. However, when PS was present, the proliferation of thymocytes co-mitogenically stimulated with PHA and exogenous IL-1 was diminished compared with the control (NHS), exhibiting its maximum inhibition when PS was added at the highest concentration and at the beginning of the incubation period. The suppressive effect of PS in the thymocyte assay disappeared with the combined action of exogenous IL-1 and IL-2. The response of T lymphocytes allogeneically stimulated was decreased in the presence of PS even when IL-1 was added; the recovery of proliferation being strictly dependent on the presence of the appropriate dose of exogenous IL-2. These results indicate that PS was interfering with IL-1 activity by rendering IL-2 producer cells unable to synthesize IL-2 and subsequently to proliferate. PMID- 2971111 TI - T/non-T and CD4/non-T cell autologous mixed lymphocyte reactions in juvenile rheumatoid arthritis. AB - Proliferation of T and CD4 cells in the autologous mixed lymphocyte reaction (AMLR) was determined for children with juvenile rheumatoid arthritis (JRA) and children with other rheumatic and connective tissue diseases. Children with musculoskeletal symptoms but no rheumatic disease and healthy adults served as controls. Patients with polyarticular rheumatoid factor (RF) positive JRA had a diminished CD4/non-T cell AMLR, whereas those with RF negative polyarticular and pauciarticular onsets had normal results. PMID- 2971112 TI - The efficacy of selective unilateral temporal artery biopsy versus bilateral biopsies for diagnosis of giant cell arteritis. AB - Two hundred bilateral temporal artery biopsies performed prospectively in 200 patients suspected of having giant cell arteritis (GCA) were always preceded by Doppler flow study and local clinical examination. If bilateral temporal artery biopsies had been guided by positive local clinical examination, 24 positive biopsies would have been found, and Doppler studies would have improved the diagnostic score to 39; however, the biopsies gave 42 histologically positive results, providing the best diagnostic yield. The possibility of GCA with normal Doppler flow studies was noted as well as the existence of relatively asymptomatic forms of GCA, so that systematic performance of bilateral temporal artery biopsy would seem justified in any case of suspected GCA. PMID- 2971113 TI - Oral zinc supplementation in Down's syndrome: restoration of thymic endocrine activity and of some immune defects. AB - Eighteen non-institutionalized Down's syndrome (DS) children (mean age: 7.0 +/- 10/12 years) with a history of respiratory tract, auditory and skin infections, low plasma levels of a nonapeptide thymic hormone, i.e. Serum Thymic Factor (STF), high plasma levels of inactive zinc-unbound STF molecules, and reduced absolute number of circulating T-lymphocytes, were given an oral non pharmacological supplementation of zinc sulphate (1 mg Zn++/kg body weight/day for 2 months; two cycles, 10 months apart) and monitored immunologically before and after each cycle. A dramatic increase of plasma STF level and concomitantly an almost complete disappearance of inactive STF molecules was observed after each cycle. The absolute number of circulating T-lymphocytes was significantly increased by zinc treatment. The marginal zinc deficiency was also corrected without any appreciable influence on copper plasma levels. A reduction of recurrent infections and an improvement in school attendance after zinc supplementation were recorded. These beneficial effects of zinc supplementation were also noted in those DS children who did not show an apparent zinc deficiency, as assessed by measuring zinc plasma level. The reduced number of circulating B lymphocytes and the impaired lymphocyte responsiveness to phytohaemagglutinin and concanavalin A were not restored. On the whole, these findings suggest that there exists a defect in the bio-availability and/or in the utilization of zinc in DS. This alteration, of unknown origin, can be underestimated on the simple basis of the zinc plasma level and can be corrected with moderate nutritional zinc supplementation. PMID- 2971114 TI - Analysis of blood cell populations, plasma zinc and natural killer cell activity in young children with Down's syndrome. AB - Down's syndrome (DS) is associated with respiratory infections, lymphoblastic leukemia and a mortality rate that is elevated particularly during the first 5 years of life. Unfortunately, most studies on immune function in DS have been performed on adults or older children, the results of which may be not relevant to a primary immune defect in DS since maturation of the immune system and compensatory mechanisms may mask to a certain degree primary immune defects which are associated with this syndrome. In the present study, several immune and hematologic parameters were studied in DS children under 6 years of age and in age-matched controls. The children with DS exhibited a reduced number of lymphocytes while the total number of white blood cells was normal. In addition, the children with DS had reduced numbers of circulating OKT4+ cells and a significantly depressed ratio of OKT4+ to OKT8+ cells, but their overall proportion of T-cells was normal. Normal NK activity was observed and cells from DS children were no more sensitive to the augmenting effects of alpha interferon than cells from control children. Hematocrit levels were slightly elevated in the DS group while plasma zinc levels appeared to be normal in this group. PMID- 2971115 TI - Parents' perception of social and sexual functions in adolescents with Down's syndrome. AB - The authors investigated parental perceptions of social interactions, interest in the opposite sex, sexual functions and issues concerning sex education in young persons with Down's syndrome. The evaluation of the data revealed that more than half of the study population showed interest in the opposite sex and are attending social gatherings. Many of the youngsters had expressed a desire to get married, however, only a few had an interest in sexual relationships. Masturbation was observed in 40% of the young men and in 22% of the young women. About half of the parents feel that their children should be sterilized or should have other forms of birth control if they would be able to reproduce. Only a limited number of adolescents had sex education. More parents who have girls with Down's syndrome than those who have boys are worried that their child may be taken advantage of sexually. PMID- 2971116 TI - Maltose transport and starch binding in phage-resistant point mutants of maltoporin. Functional and topological implications. AB - The relationships between the bacteriophage lambda binding site, the starch binding site and the pore formed by maltoporin (LamB protein, lambda receptor protein) were investigated. Bacteria with single amino acid substitutions in the maltoporin sequence, which were previously shown to be strongly reduced in phage lambda sensitivity, were assayed for maltose- (and maltodextrin) selective pore functions. Maltose transport assays was performed at low substrate concentrations, under conditions where LamB is limiting for transport. It revealed three classes of mutants. Class A is composed of mutants with no effect on transport (substitutions at amino acid residues 154, 155, 259, 382 and 401); class B corresponds to mutants with a significant but variable reduction in transport (sites 148, 151, 152, 163, 164, 245, 247 and 250); class C is represented by a single mutant for which transport is almost completely abolished (site 18). Starch binding was assayed by two different methods that gave compatible results. In class A mutants, binding was normal, while no binding was observed in the class C mutant. Binding was impaired to various extents in category B mutants. There was a correlation between the level of impairment of starch binding and impairment of maltose transport, consistent with the notion that the residues influencing starch binding are inside, or in close proximity to, the pore. These results, together with previous data on starch-binding mutants that were not affected in phage binding (substitutions at residues 8, 74, 82, 118 and 121), suggest that the binding sites for starch and phage lambda overlap but are distinct. Mutations affecting transport and starch binding are located in the first third of the protein and in the region of residues 245 to 250. Mutations affecting phage adsorption are located mainly in the last two thirds of the protein. The topological constraints suggested by the results with the available mutants altered in the lamB gene were used to propose a revised model of maltoporin folding across the outer membrane as well as to define the outlines of footprints of macromolecular binding sites (phage, starch and monoclonal antibodies) on the surface of the protein. PMID- 2971117 TI - Effect of point mutations on the in-vitro pore properties of maltoporin, a protein of Escherichia coli outer membrane. AB - Maltoporin (LamB protein), a protein of Escherichia coli outer membrane forms ionic channels with a selectivity for maltose and maltodextrins (Dargent et al., 1987). The effect of different point mutations on maltoporin pore properties was investigated in vitro with planar bilayers. The mutations belong to three classes in terms of selective maltose transport in vivo: class A (substitution at positions 259 and 382) does not affect maltose transport, class B (position 163 and 245) decreases maltose transport down to 20 to 30%, and class C (position 18) almost completely abolishes selective maltose transport. This in-vitro study reveals that class A does not affect the pore properties in contrast to class B substitutions. The class B maltoporins are still able to form channels but display some specific features and altered specificity for maltose and maltodextrins. The substitution (Gly18----Val) alters trimer stability and impedes pore function (class C mutant). Thus, there is a good correlation between the specific transport properties of the mutated maltoporins in vivo and their behavior in vitro. These data, in combination with the asymmetric orientation of the protein within the bilayer and topological considerations, indicate that residues 245 and 163 do not belong to the selectivity filter. Mutations at these sites cause hindrance at the mouth of the pore on the outer domain of maltoporin. PMID- 2971118 TI - Measurement of impotence by laser Doppler flowmetry and conventional methodology. AB - The problem of vasculogenic impotence was evaluated by laser Doppler flowmetry with the LD5000 capillary perfusion monitor to determine cutaneous penile blood flow in 25 men defined previously as impotent by Doppler ultrasound with penile brachial ratios of 0.63 +/- 0.14. Erotic visual stimulation for 1, 2 and 4 minutes did not statistically change the cutaneous flow as measured by laser Doppler flowmetry from its baseline of 20 mv. The dynamics of skin microcirculation in the penis may reflect changes in the deeper vessels and represent an alternate, dependable method to evaluate capillary perfusion. PMID- 2971120 TI - Femoropopliteal percutaneous transluminal angioplasty for limb salvage. AB - The role of balloon catheter femoropopliteal percutaneous transluminal angioplasty (FP PTA) continues to be controversial 14 years after its initial description. In high-risk patients with limb-threatening ischemia, the role of FP PTA is unknown. We reviewed the cases of 27 patients in whom high operative risk and advanced ischemia resulted in referral for PTA of the femoropopliteal segment. Technical and hemodynamic success rates were 74% and 52%, respectively. Patient follow-up was 3 to 84 months after PTA (mean 24 months). Nine patients (33%) had successful limb salvage for more than 12 months. The cumulative limb salvage rate was 47% at 2 years. Established prognostic criteria did not predict this subset of patients. Hemodynamic improvement after PTA did correlate closely with both early and late successful limb salvage. There were two periprocedural deaths. Morbidity included an acute arterial occlusion necessitating an emergency FP bypass and two minor complications. FP PTA is useful in selected high-risk patients with limb-threatening ischemia. Protocols to facilitate successful long term limb salvage must include close liaison between the vascular surgeon and interventional radiologist, intensive periprocedural monitoring, and serial noninvasive hemodynamic assessment. PMID- 2971119 TI - Long-term therapy with a depot luteinizing hormone-releasing hormone analogue (Zoladex) in patients with advanced prostatic carcinoma. AB - We treated 191 patients with histologically proved locally advanced stage (T3 or T4) and/or metastatic prostate cancer with a biodegradable depot formulation of a luteinizing hormone-releasing hormone analogue (Zoladex). After an initial increase in serum testosterone in week 1 of therapy a continuous decrease of testosterone to castrate levels was obtained. With a monthly injection of the luteinizing hormone-releasing hormone analogue 4 patients (2 per cent) experienced a transient increase in bone pain, 1 had ureteral obstruction and 1 suffered paraplegia during the first few weeks of therapy. Over-all objective and subjective responses were similar to those obtained by castration or estrogen therapy. The absence of local and systemic (long-term) side effects proves the validity of this approach for patients with advanced prostatic cancer. PMID- 2971121 TI - Results of renal artery balloon angioplasty limit its indications. AB - Percutaneous transluminal balloon angioplasty (PTA) of 83 renal artery lesions in 55 patients was done because of renal failure in eight patients, hypertension in 35, renal failure and hypertension in 11, and polycythemia in one patient. Early results in 38 patients with arteriosclerosis showed five (13%) were worse and 13 (34%) were unchanged. Twenty patients (53%) with arteriosclerosis were initially cured or improved; however, seven of these patients had recurrence in 4 to 48 months. Ultimately, only 13 of 38 patients with arteriosclerosis (34%) were cured or improved (mean follow-up 22 months). Nine patients with fibromuscular dysplasia required 17 dilatations of arteries (three bilateral and five repeat), resulting in eight patients (89%) who were cured or improved. Selection of patients with hypertension by medical failure while receiving three or more hypertension medications or by lateralizing renal vein renin values yielded benefit in 17 of 26 patients (65%). Five of six patients with transplant stenosis of the renal artery and hypertension were cured or improved at mean follow-up of 18 months. Overall technical results of 83 artery dilatations were as follows: good, 58 (69%); fair, 10 (12%); poor or unsatisfactory, 16 (19%); these were judged with a blinded radiologic review. No patient suffered main renal artery thrombosis. There were 16 patients with complications of dilatation (morbidity rate of 29%). Nine patients subsequently had renal artery surgery from the same day to 64 days later with good results in all patients except one.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971122 TI - [Three cases in a family of congenital protein S deficiency associated with cerebral infarction]. PMID- 2971123 TI - [Various types of glomerulonephritis. Minimal change glomerulonephritis]. PMID- 2971124 TI - [Clinical experience of PTCA for ischemic heart disease]. PMID- 2971125 TI - [Recent trends in occupational skin disorders]. PMID- 2971127 TI - [Possibilities of laparoscopy in the diagnosis of disseminated cancer of the stomach]. PMID- 2971126 TI - [Early bypass grafting following intracoronary thrombolysis in acute myocardial infarction]. PMID- 2971128 TI - [Causes of the delayed detection of cancer of the colon]. PMID- 2971129 TI - [Roentgeno-endovascular revascularization of organs and tissues]. PMID- 2971130 TI - [Combined treatment of acute obstruction of major arteries]. PMID- 2971132 TI - [Occupational cold-induced polyneuropathy]. PMID- 2971131 TI - Impaired renal responsiveness to human atrial natriuretic peptide (hANP) in normotensive patients with type 1 diabetes mellitus. AB - Potential impairment of the efficacy of human atrial natriuretic peptide (human ANF-(99-126), hANP), the most potent endogenous natriuretic agent in healthy subjects, was examined in eight male normotensive patients with uncomplicated type 1 diabetes mellitus (aged 22-37 years). After giving informed consent, patients and eight male control subjects (aged 22-28 years) received in a random double-blind study design i.v. bolus injections of 100 micrograms hANP (Bissendorf peptide) or placebo. At base-line, patients differed from controls in elevated creatinine clearance (P less than 0.05) and in mild postprandial hyperglycemia. Whereas the responses of urinary cyclic guanosine monophosphate, the second messenger of hANP, were found to be normal in patients, the diuretic and natriuretic effects of hANP were grossly impaired when compared to controls (P less than 0.01); hANP resulted in increased plasma protein concentrations only in controls (P less than 0.05 vs patients). In both groups, creatinine clearance remained uninfluenced by hANP. There were similar decreases in plasma renin activity, aldosterone, levels, and blood pressure (systolic more than diastolic) in both groups (P less than 0.05 vs placebo). Heart rate and blood glucose remained unchanged. Thus, there is evidence for a decreased responsiveness to hANP exclusively of renal fluid, sodium, and chloride excretion in uncomplicated type 1 diabetes mellitus. The mechanisms responsible for this phenomenon remain obscure, neither a down regulation at the hANP receptor sites nor an hANP-induced shift from intra- to extravascular fluid volume are likely to be involved in its probably diabetes-specific pathogenesis. PMID- 2971133 TI - [Equipment for the rehabilitation of patients]. PMID- 2971134 TI - The role of capsule as a barrier to bacteriophage adsorption in an encapsulated Staphylococcus simulans strain. AB - The polyvalent staphylococcal bacteriophage U16 failed to adsorb to an encapsulated Staphylococcus simulans strain. Partially purified cell wall and teichoic acid of this strain could, however, inactivate bacteriophage U16 to a great extent, indicating the presence of the phage receptor. It is concluded that the capsule of Staphylococcus simulans acts as a barrier for the interaction of the phage with its receptor in the bacterial cell wall. PMID- 2971135 TI - Interleukin-1 directly regulates hormone-dependent human breast cancer cell proliferation in vitro. AB - Direct in vitro effects of IL-1 on hormone-dependent (MCF-7 and ZR-75-B) and independent (HS-578-T and MDA-231) human breast cancer cell proliferation were investigated in short-term and long-term cell cultures. For short-term (48 h) studies [3H]thymidine uptake was used as an index of proliferation, while for long-term (12 day) cultures actual cell numbers were determined. Initial studies, conducted with MCF-7 cells, demonstrated that both forms of recombinant human IL 1 (alpha and beta) at 10(-11) M inhibited [3H]thymidine uptake by MCF-7 by 70%, and by day 7 of the long-term study alpha and beta IL-1 at 10(-11) M inhibited MCF-7 cell growth by 80%. IL-1, while inhibiting the growth of another hormone dependent breast cancer cell line; ZR-75-B, had no effect on the hormone independent cell lines MDA-231 and HS-578-T. The differing proliferative responses of the hormone-dependent and independent cells to IL-1 may, in part, be due to the expression of IL-1 receptors on these cells, in that MCF-7 cells express IL-1 receptors [dissociation constant (Kd) = 2.0 x 10(-10) M; receptor density = 2,500 sites per cell and mol wt = 80,000] while the hormone-independent MDA-231 cells do not. PMID- 2971136 TI - The interaction of IgG subclasses with solubilized Fc receptors of rat basophilic leukemia cells. AB - Rat basophilic leukemia (RBL) cells carry two surface glycoprotein molecules named R (or alpha) and H which, when detergent solubilized, bind to rat IgE Sepharose. The same two molecules also bind to rat IgG-Sepharose but with a lower affinity. R is a component of the high affinity Fc receptor for IgE. In the present study the inhibition of the binding of R and H to rat IgG-Sepharose by various homologous and heterologous immunoglobulins was used to assess their relative affinities for the two receptor molecules. Ranking the rat immunoglobulins in order of their affinities for the R receptor yielded: IgE much greater than IgG2a greater than IgG1 greater than IgG2b; and for H: IgE greater than IgG2b greater than IgG1 greater than IgG2a. Rat IgG2c inhibited the binding of both R and H but a precise ranking could not be assigned. Conclusive evidence has been obtained for the Fc specificity of these interactions. The affinities of the mouse IgG subclass/R interactions can be ranked: IgG1 greater than IgG2a greater than IgG2b; and for the H receptor: IgG1 greater than IgG2b greater than IgG2a. All of the mouse proteins and other heterologous IgGs, such as those of sheep, goat, equine and rabbit origin, interacted considerably more strongly with H than with R. No interaction with mouse IgG3 could be detected under the conditions tested. PMID- 2971137 TI - Studies with a monoclonal antibody to the beta subunit of the receptor with high affinity for immunoglobulin E. AB - The receptor with high affinity for IgE consists of a tetrameric complex of polypeptides, one of which (alpha), contains the binding site for IgE. The function of the other chains--a single beta and two disulfide-linked gamma chains -is unknown. We report the cloning of a murine hybridoma that secretes an IgG1 antibody which specifically reacts with the beta subunit. Studies with this monoclonal antibody show that the subunit stoichiometry of the receptor is unaffected by the presence or absence of bound IgE. We also found that under certain conditions where the alpha beta gamma 2 complex dissociates, beta remains attached to the dimer of gamma chains, indicating that these chains contact each other in the native receptor. In rat basophilic leukemia cells--a neoplastic line of mucosal-type mast cells--all of the beta subunits expressed by the cells appeared to be associated with the high affinity receptor. However, in at least one cell line which has no high affinity receptors--a putative rat lymphoma line- beta or beta-like polypeptides were also expressed. PMID- 2971138 TI - X-ray-induced specific locus mutations in the ad-3 region of two-component heterokaryons of Neurospora crassa. I. Modification of the heterozygous effects of multilocus deletions covering the ad-3A or ad-3B loci. AB - The basis for the reduced growth rates of heterokaryons between strains carrying nonallelic combinations of gene/point mutations (ad-3R) and multilocus deletion mutations (ad-3IR) has been investigated by a simple genetic test. The growth rates of forced 2-component heterokaryons (dikaryons) between multilocus deletion mutations were compared with forced 3-component heterokaryons (trikaryons) containing an ad-3AR ad-3BR double mutant as their third component. Since the third component has no genetic damage at other loci immediately adjacent to the ad-3A or ad-3B locus, the growth rate on minimal medium depends on the functional activity of the unaltered (and presumed "wild-type") ad-3A and ad-3B loci in the first two components. In many cases, the requirements of the original dikaryons have been satisfied by the addition of unaltered genes (in the third component), and these trikaryons grow at wild-type rate on minimal medium. Those trikaryons growing at less than wild-type rate were shown to be adenine-requiring, and wild type growth rate was obtained with the addition of low levels of adenine to the medium. Such tests in the present experiments have shown that ad-3IR mutations result not only in inactivation of the ad-3 loci by multilocus deletion but also, in many cases, in partial gene inactivation by an unknown mechanisms at other loci in the immediately adjacent regions. The heterozygous effects observed in our present experiments with multilocus deletions in Neurospora can be explained either by a spreading-type position effect of the type found by others in Drosophila, mice, Oenothera and Aspergillus or by undetected genetic damage ("cryptic mutations") in the immediately adjacent genetic regions. An attempt will be made to distinguish between these two alternative hypotheses with techniques for DNA cloning and sequencing in future experiments. PMID- 2971139 TI - Abnormal red-cell calcium pump in patients with idiopathic hypercalciuria. AB - Idiopathic hypercalciuria is a common disorder whose inheritance suggests an enzyme abnormality in calcium transport. We measured calcium-magnesium-ATPase activity in erythrocytes from 38 patients (mean age [+/- SEM], 40 +/- 2.1 years) with idiopathic hypercalciuria (24-hour urinary calcium excretion greater than or equal to 0.1 mmol per kilogram of body weight) and a history of multiple calcium oxalate kidney stones. As compared with 41 healthy controls, the patients with hypercalciuria had increased erythrocyte-membrane calcium-magnesium-ATPase activity (64.2 +/- 2.19 vs. 51.6 +/- 1.91 nmol of ATP split per milligram per minute; P less than 0.01) and increased sodium-potassium pump activity (6866 +/- 233 vs. 6096 +/- 228 mumol of sodium per liter of red cells per hour; P less than 0.05). No significant difference between the two groups was found in erythrocyte sodium-potassium cotransport, sodium-lithium countertransport, or potassium content. In 66 patients with kidney stones (38 with hypercalciuria and 28 with normal calcium excretion), 24-hour urinary calcium excretion correlated with calcium-magnesium-ATPase activity (r = 0.46, P less than 0.001). Erythrocyte calcium-magnesium-ATPase activity remained unchanged in eight subjects studied after four months on a low-calcium diet. A study of 30 healthy families found significant correlations between mean values in parents and those in offspring for calcium-magnesium-ATPase (r = 0.68, P less than 0.001) and urinary calcium excretion (r = 0.45, P less than 0.02), with no significant correlations between parents with respect to these measures (r = 0.27 and r = 0.08, respectively). We conclude that abnormalities in erythrocyte calcium-magnesium-ATPase activity may represent an inherited defect in calcium transport related to the cause of idiopathic hypercalciuria. PMID- 2971140 TI - Corn as a source of mycotoxins in Indonesian poultry feeds and the effectiveness of visual examination methods for detecting contamination. AB - Every truck load of corn (n = 52) entering and every batch of poultry feed (n = 290) leaving a Bogor feedmill over one year was analysed for aflatoxins, zearalenone, ochratoxin A and sterigmatocystin. Fifty loads of corn and 274 of the batches of chicken feed contained aflatoxins. Zearalenone was detected in 11 corn samples but was not found in the formulated feed. Ochratoxin A was detected in one corn sample, but not in feed. Corn can account for all of the aflatoxin in the feed since levels were always lower in the finished product. There was no quantitative association between the proportion of bright green-yellow fluorescent, purple or mouldy kernels and the mycotoxin contents of the composite samples. Nevertheless, the absence of abnormal kernels indicates higher quality corn since the highest levels of mycotoxins occurred in the abnormal kernels. PMID- 2971141 TI - Function of c-mos proto-oncogene product in meiotic maturation in Xenopus oocytes. AB - The c-mos proto-oncogene is expressed as a maternal mRNA in oocytes and early embryos of Xenopus laevis, but its translation product pp39mos is detectable only during progesterone-induced oocyte maturation. Microinjection of mos-specific antisense oligonucleotides into oocytes not only prevents expression of pp39mos, but also blocks germinal vesicle breakdown, indicating that it functions during reinitiation of meiotic division. PMID- 2971142 TI - Trans splicing involves a novel form of small nuclear ribonucleoprotein particles. AB - The trans-splicing reaction occurring in trypanosomes and related species as well as in the nematode Caenorhabditis elegans involves the transfer of a 5' exon from a spliced leader transcript (SL RNA) to a precursor messenger RNA transcript with a 3' splice acceptor site. This seems to take place in the same nuclear compartment as normal cis splicing and proceeds through Y-branched intermediates analogous to the lariats formed in cis splicing. The cellular machinery catalysing cis and trans splicing might therefore be expected to share some components, particularly in the nematode where some mRNAs are produced by both cis and trans splicing. We generated possible secondary structures for the SL RNAs of several species and found they were remarkably similar although neither nucleotide sequence nor length is conserved. Each contained three stem-loops; strikingly the 5' splice site is adjacent to the turn of the most 5' loop and an Sm-binding consensus sequence is found between the second and third stem-loops. Sm is an antigen associated with small nuclear ribonucleoprotein particles (snRNPs). When incubated in HeLa cell nuclear extracts, SL RNAs become immunoprecipitable by anti-Sm, but not by other autoantibodies directed against proteins of mammalian snRNPs. We propose that SL RNAs have a dual function in the trans splicing process: they consist of a 5' exon covalently linked to an snRNA like sequence and seem likely to exist as Sm snRNP particles (SL snRNPs) within the cell. Just as the RNA in the U1 snRNP base-pairs with the 5' splice site, rendering it susceptible to attack in the cis-splicing reaction, so might the SL snRNP autonomously activate its own 5' splice site and thereby eliminate the need for a U1-like snRNP in the trans-splicing machinery. PMID- 2971143 TI - Partial agonists of the morphine type: facilitative effects on electrophysiologically recorded C-fiber reflexes in the cat. AB - The purpose of the present study was to examine the effects of the partial agonists of the morphine type, buprenorphine, profadol and propiram on the spinally-mediated C-fiber reflex. Buprenorphine, profadol and propiram facilitated the C-fiber reflex. In the presence of naltrexone, the effects of buprenorphine were changed to depression of the C-fiber reflex. Profadol and propiram continued to facilitate the C-fiber reflex in the presence of naltrexone. The present results suggest that buprenorphine facilitated the C fiber reflex by blocking endogenous opioid peptide inhibition of the C-fiber reflex whereas profadol and propiram facilitated it by a non-opioid mechanism. PMID- 2971144 TI - The D1 dopamine receptor in the rat brain: quantitative autoradiographic localization using an iodinated ligand. AB - The distribution of dopamine D1 receptors in the rat, labeled with [125I]SCH 23982, was studied using a quantitative in-vitro light-microscopic autoradiographic method. The binding of [125I]SCH 23982 to slide-mounted tissue sections and membrane preparations of prefrontal cortex was saturable, specific and of high affinity. Scatchard analysis revealed a Kd of 1.15 +/- 0.47 nM and Bmax of 8.76 +/- 0.34 fmol/mg tissue in prefrontal cortex membranes and a Kd of 1.27 +/- 0.14 nM and Bmax of 67.6 +/- 3.75 fmol/mg tissue in slide-mounted tissue sections at the level of the striatum. [125I]SCH 23982 was found to predominantly label D1 receptors, but a small fraction of the binding was to serotonin receptors. D1 receptors were found throughout the forebrain and were concentrated in the substantia nigra pars reticulata, accumbens nucleus, caudate putamen, entopeduncular nucleus, olfactory tubercle and the major island of Calleja. [125I]SCH 23982 binding to serotonin receptors was concentrated in the cortices, dorsal raphe, central gray, anterior hypothalamic area and the molecular cell layer of the cerebellum. Knowledge of the distribution of D1 receptors may increase our understanding of the role of D1 receptors in central nervous system dopaminergic function. Furthermore, data on the potential sites of interaction of [125I]SCH 23982 with serotonin receptors may help to understand the complex physiology and pharmacology of the primarily D1 selective compound. PMID- 2971146 TI - Back pain. PMID- 2971145 TI - Vascular recanalizing techniques in the hind brain circulation. AB - Percutaneous transluminal angioplasty (PTA) was performed in 45 patients with a manifest subclavian steal syndrome. Thirty-five of those patients were subjected to follow up examinations over a period of 6 to 18 months. Five patients suffered from severe restenosis and were treated again. Two thirds of the patients benefited from the treatment. PTA of the proximal vertebral artery was performed in 15 patients with bilateral occlusive lesions of the extracranial vertebral arteries. In 13 of these cases the neurological and the vascular states of the patients were regularly reexamined, 8 showed a marked improvement. During the 2 to 25 month observation period (average 15 months post-PTA) reocclusion was observed in only two cases. These showed no recurrent neurological sequelae. Forty-three consecutive patients with acute vertebro-basilar or basilar occlusion received intraarterial fibrinolytic therapy with streptokinase or urokinase. Twenty-three of these had presented severe deficits at the beginning of therapy (e.g. complete tetraplegia, comatous state for more than 6 hours). None of this group survived. By contrast the 20 other patients in this group presented with incomplete fluctuating or progressive motor deficits. None was comatous for more than 6 hours. Fourteen patients (33% in this group) survived. Local intraarterial fibrinolytic therapy is the only therapy successful in the treatment of progressive stroke from vertebro-basilar thrombosis. PMID- 2971147 TI - Circulatory congestion in obese hypertensive women: a subset of pulmonary edema in pregnancy. AB - We describe four obese, chronically hypertensive women presenting with antepartum pulmonary edema in whom invasive hemodynamic monitoring showed elevated wedge pressure, normal to high cardiac index, and normal systemic vascular resistance. Echocardiography revealed large chambers, thick walls, and increased left ventricular mass with normal systolic but abnormal diastolic function. These findings are indicative of intrinsic volume overload occurring in the presence of impaired left ventricular relaxation, a combination resulting in high filling pressures and pulmonary congestion. Diuretic therapy is indicated in this subset of patients, who could not be recognized by the usual clinical parameters such as history and physical examination, chest x-ray, and arterial blood gas. PMID- 2971149 TI - [Early clinical manifestations of spondylolisthesis in children]. PMID- 2971148 TI - Prostaglandin E2-mediated suppression of human maternal lymphocyte alloreactivity by first-trimester fetal macrophages. AB - We examined the immunosuppressor role of the first-trimester human placental macrophages on maternal lymphocyte alloreactivity in vitro to determine whether these macrophages and their secreted mediator(s) participate in the local immunoregulation at the fetomaternal interface. These cells were tested for their effects on maternal lymphocyte proliferation when added alone (as stimulator cells) or as regulator cells in 1) a one-way mixed lymphocyte reaction for six days and 2) lymphocyte cultures in the presence of phytohemagglutinin for three days. We detected a dose-dependent inhibition of phytohemagglutinin-induced proliferation with 48-hour macrophage culture supernatants. Placental macrophages, when used alone as stimulator cells in the mixed lymphocyte reaction, were stimulatory only at a concentration of 1-2%; the stimulation was abolished at a concentration of 10%. When used as regulator cells, they showed a strong inhibition of mixed lymphocyte reaction and phytohemagglutinin-induced proliferation at a concentration of 10%; no effect was observed at a concentration of 1-2%. The major class of the molecules mediating the suppression was identified as prostaglandins, primarily prostaglandin E2 (PGE2), based on the findings that the presence of indomethacin (10(-5) M) or various dilutions of an anti-PGE2 antibody abrogated suppression substantially or completely. Prostaglandin E2 levels measured in the mixed leukocyte culture wells containing placental macrophages as regulator cells correlated positively with the macrophage dose and its suppressive effect. PMID- 2971150 TI - [An orthopedic chair on rollers with a rotating seat for patients and invalids]. PMID- 2971151 TI - [A mathematical model of the prognosis of trauma among children]. PMID- 2971152 TI - Factors affecting the normal perimetric profile derived by automated static threshold LED perimetry. I. Pupil size. AB - The relationship between pupil size and perimetric sensitivity was investigated in a group of clinically normal emmetropic subjects using the Dicon AP3000 computer-assisted perimeter at bowl luminances of 10 asb and 45 asb. Pupil size was modified with the miotic thymoxamine 0.5% and the mydriatic phenylephrine 10%; saline was used as the control. Perimetric sensitivity increased with an increase in pupil size. The effect was greatest for peripheral locations for both bowl luminances. The fluctuations in perimetric sensitivity increased with peripheral angle and with decrease in pupil size for both bowl luminances. PMID- 2971153 TI - Expectancies and functional impairment in chronic low back pain. AB - Perceived self-efficacy [2] and pain response expectancies [13] were examined as correlates of movement limitations and impaired functioning in a sample of 40 patients with chronic low back pain. Self-efficacy was operationalized as patients' ratings of their ability to perform movements, while response expectancies were operationalized as ratings of the degree of pain expected to accompany the movements. Patients predicted their ability to perform 10 simple movements and the degree of pain that would accompany each movement; on a separate occasion, subjects' actual performance of the movements was assessed. In general, independent predictions of both movement ability and pain correlated significantly with performance of specific movements as rated from videotapes. Total movement scores correlated 0.55 and -0.54, respectively, with total self efficacy and pain response expectancy ratings. Expectancy ratings were also predictive of average daily pain ratings and questionnaire ratings of physical impairment in everyday life. Multiple regression analyses, including causal modeling, indicated that actual performance was best predicted by self-efficacy ratings, which in turn appeared to be determined by pain response expectancies. PMID- 2971154 TI - Haemodynamic actions of clonidine in tetraplegia--effects at rest and during urinary bladder stimulation. AB - We studied the haemodynamic effects of clonidine (2 micrograms/kg/iv) in 7 tetraplegics and 7 normal subjects. Measurements of blood pressure, stroke volume, cardiac output and digital (finger) skin blood flow were made before and after clonidine for 60 minutes. Blood pressure, stroke volume and cardiac output did not fall in tetraplegics, unlike normals. Resting digital skin blood flow was higher in tetraplegics and fell after clonidine. In normal subjects however, an increase in digital skin blood flow occurred after clonidine. The pressor and digital vasoconstrictor responses to bladder stimulation were attenuated after clonidine. The inability of clonidine to induce a fall in blood pressure, stroke volume, cardiac output and cause peripheral vasodilation in tetraplegics is consistent with its central sympatholytic effects. Attenuation of the responses to bladder stimulation suggest an effect on spinal sympathetic neurones. PMID- 2971155 TI - Mechanisms for the anticoagulant effect of heparin and related polysaccharides. AB - The anticoagulant actions of heparin and related polysaccharides can best be addressed by answering the following 2 questions: to what extent do unfractionated heparin, low molecular weight heparins, dermatan sulfate and heparan sulfate inhibit the activation of prothrombin in plasma depleted of antithrombin III and heparin cofactor II? What roles do antithrombin III and heparin cofactor II play in the expression of the overall anticoagulant effects of heparin, low molecular weight heparins, dermatan sulfate and heparan sulfate? Since only heparin can, but only weakly at best, inhibit the activation of prothrombin in plasma depleted of both antithrombin III and heparin cofactor II, it is obvious that the anticoagulant effects of heparin, low molecular weight heparins, dermatan sulfate and heparan sulfate are mediated primarily by their catalytic effects on the antiprotease actions of antithrombin III (heparin, low molecular weight heparins and heparan sulfate) and heparin cofactor II (dermatan sulfate). The catalytic efficiencies of various glycosaminoglycans (GAGs) on the inhibition of thrombin by undiluted plasma follow in the order of the ability of each GAG to inhibit the intrinsic pathway activation of prothrombin. The reasons for this observation probably follow from the effects of GAGs on the 2 positive amplification reactions of thrombin during blood coagulation. Factor VIIIa and factor Va, which directly or indirectly contribute to the rapid formation of prothrombinase, arise from the limited proteolysis of factor VIII and factor V by thrombin and/or factor Xa. On depletion of thrombin by enhanced formation of thrombin-antithrombin III or thrombin-heparin cofactor II, factor Xa provides the only mechanism by which factor VIII and factor V activation in plasma will proceed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971156 TI - The sequence of the yeast ATP5 gene. PMID- 2971158 TI - Suppression of unexpected recombinant background in lambda EMBL3 cloning. PMID- 2971159 TI - New derivatives of lambda replacement vectors EMBL3 and EMBL4. PMID- 2971157 TI - Human U1 snRNP-specific C protein: complete cDNA and protein sequence and identification of a multigene family in mammals. AB - A complementary DNA clone for the human U1 snRNP-specific C protein has been isolated. The nucleotide sequence of the 733 bp cDNA insert includes a 15 bp 5' untranslated region, an open reading frame of 477 bp corresponding to 159 amino acids (Mr = 17,373 D), and a 223 bp 3'-untranslated region. The identity of the clone was confirmed by in vitro translation of hybrid-selected mRNA or an RNA transcript synthesized from the cDNA. The in vitro synthesized C protein has a slightly greater mobility on SDS-polyacrylamide gels, indicating that the in vivo product is post-translationally modified. The deduced primary structure contains a segment of high proline and methionine content. A region homologous to the RNP consensus sequence, found in the other two U1 snRNP-specific proteins 70K and A, is absent. Analysis of genomic DNA restriction enzyme digests shows hybridizing fragments in the genome of all vertebrate classes. The results are consistent with multi-copy representation of the C protein gene in mammals, whereas in the other vertebrate classes the related protein seems to be encoded by a single-copy gene. PMID- 2971161 TI - Midwives' Journal. PMID- 2971160 TI - Charon BS(+) and (-), versatile lambda phage vectors for constructing directional cDNA libraries and their efficient transfer to plasmids. PMID- 2971162 TI - [Why vaccines from genetic engineering?]. PMID- 2971163 TI - [Hepatic encephalopathy]. PMID- 2971164 TI - [Study of the hyo-thyro-epiglottic space by ultrasonography]. AB - The pre-epiglottic space is located superficially at the junction of the larynx, oropharynx and hypopharynx. When it is invaded by a neighbouring cancer, the latter rises to stages 3 or 4 of the TNM classification. The space is explorable by percutaneous ultrasonic scanning of the neck. The present paper gives the first description of the normal sonographic anatomy of the pre-epiglottic space, based on 16 healthy volunteers. It also gives the first description of invasion of that space, based on 18 patients operated upon for ENT (ear, nose and throat) cancers. In 5 of these 18 patients, invasion of the pre-epiglottic space was proven at histology. There were 3 positive and no false negative results, but 3 false positive results were obtained at the beginning of the series. The accuracy of ultrasounds in this study was 83 per cent. PMID- 2971165 TI - [Renal failure disclosing interstitial granulomatous nephropathy of sarcoidosis. 4 cases]. AB - In patients with interstitial granulomatous sarcoid nephritis, renal failure often suggests the diagnosis, being rapidly progressive and isolated, i.e. without proteinuria and with few abnormalities of the urinary sediment. The extra renal signs are usually discreet or absent, and the finding of a paradoxically high or normal calcaemia concomitant with renal failure should alert the clinician. When granulomatous lesions of the kidney are not too old, corticosteroids are remarkably effective in improving renal function. PMID- 2971166 TI - [Treatment of hyperthyroidism by radioactive iodine. Development of methods and difficulties in the surveillance]. AB - The experience of treatment of hyperthyroidism with radioactive iodine in the Nuclear Medicine department of the J. Godinot Institute, Rheims, from 1967 till 1987, is described. One thousand one hundred and fifty patients (723 with diffuse and 427 with nodular hyperthyroidism) received a total of 1,565 doses. The dosage calculation method was considerably simplified, enabling a patient to be treated in 24 hours. Thyroid gland mass evaluation, initially based on the scintigraphic projection area, is now performed by ultrasonography. The mean total radioactivity administered is 370 +/- 320 MBq per patient. All patients are seen on the 8th post-treatment week for clinical examination and hormone control. Seven out of 10 patients are cured with a single dose. Long-term follow-up, based on a computer file, is effected by means of an annual letter sent to the patient and to his family doctor, but in spite of sustained efforts 36 per cent of the patients are lost sight of. The diagnosis of iodine 131-induced hypothyroidism is based on plasma levels of TSH which, since 1984, are measured by the ultrasensitive method. The overall incidence of hypothyroidism is 6.6 per cent in the first year and 3 per cent thereafter. Despite a simplified procedure, the results obtained by the authors are similar to those found in the literature, and the cost-efficiency ratio is excellent. PMID- 2971167 TI - [Detection of anti-double stranded DNA antibodies of IgG, IgA and IgM classes by ELISA. Comparison with the Farr test and indirect immunofluorescence]. AB - Anti-double stranded DNA antibodies were measured by an immunoglobulin class specific immunoenzymatic assay (ELISA), in 450 sera from 265 patients as well as by indirect immunofluorescence using Crithidia luciliae as a substrate and, for 124 sera, by the Farr test. ELISA proved specific and reproducible and it yielded results that were well correlated with the Farr assay, with a slightly higher sensitivity of ELISA. Correlation with immunofluorescence was not as good because of the lower sensitivity of the latter method. ELISA enables the levels and isotypes of anti-DNA antibodies to be determined. Both appear to be critical parameters for a clinical interpretation of results, especially with respect to the diagnosis of systemic lupus erythematosus. PMID- 2971168 TI - [Newborn infants of drug-dependent mothers]. AB - The problems of embryo-foetopathy associated with drug-dependent mothers were determined from published data and from a series of 60 neonates born of heroin addicted mothers. Low weight at birth resulting from prematurity and/or hypotrophy was frequent. Monitoring of labour and the presence of a specialized medical team in the delivery room improve the Apgar's score and reduces the incidence of meconium inhalation. The possible occurrence of seldom severe withdrawal syndrome makes it mandatory to place every child born of a drug dependent mother under observation in a neonatology unit. The predominant threat, at mid- or long term, is human immunodeficiency virus infection. The fear of AIDS explains why such children are now followed up for longer periods, resulting in a more accurate evaluation of their future somatic and psychomotor status. PMID- 2971169 TI - [Sleeping habits in Sahelian Africa]. PMID- 2971170 TI - [A rare cause of rectal bleeding: cancer of the prostate]. PMID- 2971171 TI - [Allergenicity of house dust]. PMID- 2971172 TI - [Aseptic rheumatismal osteitis associated with palmoplantar pustulosis. Value of colchicine]. PMID- 2971173 TI - [Bone metastases disclosing carcinoid tumor of the pancreas]. PMID- 2971174 TI - [Adenocarcinoid of the appendix. A case]. PMID- 2971175 TI - [Cardiogenic shock status with conductive disorders in severe poisoning with viloxazine. A case]. PMID- 2971177 TI - [RU 41740 and recurrent respiratory infections]. PMID- 2971176 TI - [Chronic primary polyradiculoneuritis. Foci of cerebral demyelinization visible in magnetic resonance imaging]. PMID- 2971178 TI - [Structure and distribution of RU 41740]. AB - RU 41740 is obtained from Klebsiella pneumoniae by organic extraction. It is mainly composed of carbohydrates with a small portion of proteins. Its active principle consists of two repetitive glyco-protein subunits: P1 which has a relative molecular mass of 95 kD and comes from the bacterial capsule, and F1 (relative molecular mass 350 kD) which is issued from the external bacterial membrane. Animal studies performed with tritiated RU 41740 have shown that the compound crosses the intestinal wall and that it rapidly and preferentially enters the lymphatic system. After oral administration to mice, the drug is detected by immunofluorescence in the basal end of the cells and chorion of the digestive tract epithelium. PMID- 2971179 TI - [Experimental immunopharmacology of RU 41740]. AB - RU 41740 is a biological immunomodulator which stimulates phagocytic cells, mostly by increasing their functional activities: cell metabolism, chemotaxis and secretion of mediators, notably interleukine 1. This compound also stimulates cellular immunity, as shown by the increase observed in proliferative responses in vitro, cell-mediated lymphocytotoxicity and natural killer cell-dependent cytotoxicity. Finally, RU 41740 stimulated B cells through a mitogenic effect and through a non-specific increase in antibody secretion. PMID- 2971180 TI - [Effect of an immunomodulator, RU 41740, on experimental infections]. AB - RU 41740 is an immunomodulator of organic origin acting on cells of the immune system (B cells, T cells, phagocytic cells) and on mediators IL1-CSF). Its mode of action has been explored by means of experimental infections. The types of defence involved differ according to whether the experimental infection is caused by an extracellular or intracellular micro-organism. Candida albicans and Saccharomyces cerevisiae were used to produce fungal infections, while bacterial infections were obtained with Staph. aureus, E. coli, Strep. pneumoniae and other organisms. The influenza virus was used to produce a viral infection. In these experimental models, RU 41740 increased the survival time of the infected animals and reduced bacterial, fungal and viral proliferation. These effects were observed even in immunocompromised mice. These in vitro and in vivo studies have demonstrated that RU 41740 is effective whatever the type of body defence involved. PMID- 2971181 TI - [RU 41740: mode of action on the respiratory system]. AB - Alveolar macrophages are issued from circulating monocytes and stand at the cross road of all antixenic defences of the lung. They are obtainable for in vitro studies by alveolar lavage, and their properties have been determined during the last few years. Synthesis activity and secretion of inflammation mediators, enzymes, enzyme inhibitors, free radicals and factors of cell co-operation are responsible for the defence of the lungs. The phagocytic activities of alveolar macrophages and their ability to promote cell co-operation make them cells that are capable of antigenic presentation, a fundamental step in the development of immunological defence mechanisms. The effectiveness of RU 41740 in respiratory infections is due to its action on deep lung cells, and in particular alveolar macrophages. This compound has been shown to recruit cells (which is a sign of macrophage activation), to increase enzymatic activities, phagocytosis and release of free radicals and to promote interleukine 1 secretion. These activities have been demonstrated in in vitro studies as well as in animal and human studies. PMID- 2971182 TI - [Clinical immunopharmacology of RU 41740]. AB - The clinical immunopharmacology of RU 41740 was studied in man by means of double blind, drug versus placebo trials. When the compound was administered concomitantly with an anti-influenza vaccine, the anti-influenza antibody titers were higher than in the group which received a placebo. RU 41740 increased delayed hypersensitivity as demonstrated in vitro by lymphocyte proliferation tests in treated subjects and in vivo by cutaneous reactions in patients with malignant diseases. The drug was also found to stimulate phogocytosis in patients with chronic bronchitis. These studies show that RU 41740 has a non-specific enhancing effect on humoral and cellular immune responses and on phagocytic functions. PMID- 2971184 TI - [Preventive action of an immunomodulator on respiratory infections in elderly subjects]. AB - Three hundred and fourteen elderly subjects admitted to chronic medical centers were given either RU 41740 (n = 155) or a placebo (n = 159) at the rate of one course per month during three months. RU 41740 was administered in doses of 2 mg per day during 8 days in the first course and 1 mg per day during 8 days in the second and third courses. The subjects were followed up and regularly examined every three months for one year. The incidence of acute infectious episodes was evaluated in both groups. Compared to those patients who received the placebo, the number of subjects without infection was significantly higher in the treated group during the 0-6 months and the 0-9 months periods and during the 12 months of observation. The number of infectious episodes was reduced during the 0-3 months and 0-9 months periods and throughout the 12 months of the trial. The mean duration of pulmonary infections that occurred during the 0-6 and 0-9 months periods was reduced. Finally, there was a significant decrease in the duration of antibiotic therapy during the 0-3, 0-6, 0-9 months periods and during the 12 months of observation. The drug was well tolerated. This study showed that RU 41740 is effective in protecting elderly and therefore fragile subjects against respiratory infections. PMID- 2971185 TI - [Prevention of infections of the respiratory tract in children with RU 41740]. AB - Several double-blind drug versus placebo trials have been carried out to evaluate the effectiveness of RU 41740 in the treatment of recurrent respiratory infections in children. The compound was administered in doses of two tablets per day for one week during the first month and one tablet per day for one week during the second and third months. Effectiveness was assessed 1, 3 and 6 months after the end of the treatment period. These trials have shown that RU 41740 is well tolerated both clinically and biochemically and that it decreases the number, duration and complications of respiratory infections, as well as the consumption of antibiotics during the infectious episodes. It therefore appears that this compound is particularly useful to prevent recurrent respiratory infections in children. PMID- 2971183 TI - [Chronic bronchitis. Value of RU 41740]. AB - Chronic bronchitis is responsible for 20,000 deaths per annum in France, i.e. 5 per cent of the overall mortality rate. Infection of the bronchi and lung tissue is a frequent cause of death in these patients. Acute on chronic bronchitis ranks fifth among the causes of disablement and admission to hospital. Pneumococci and Haemophilus influenza are the organisms most frequently isolated. the incidence and potential severity of acute episodes of infection account for the repeated use of antibiotics which carries a risk of promotion bacterial resistance. RU 41740 is a non-specific immunomodulator agent which reinforces the non-specific means of the respiratory tract against infections. Three double-blind, drug versus placebo and therefore reliable therapeutic trials have shown that the drug is effective in preventing airway infection. In patients with moderately advanced chronic bronchitis, RU 41740 reduces the number and duration of acute infectious episodes as well as antibiotic consumption. This positive effect persists in patients with chronic respiratory failure, including those who present with extensive bronchial dystrophy. RU 41740 is particularly effective in patients with numerous previous episodes of infection, but it also acts at all stages of chronic bronchitis. PMID- 2971186 TI - [Administration of RU 41740, a preventive anti-infective immunomodulator in an acute respiratory episode. Synthesis of 3 clinical trials]. AB - In both adults and children RU 41740 exerts an immunomodulating effect and prevents recurrent respiratory infections. Patients with such infections frequently consult for acute episodes, and it was deemed necessary to evaluate the safety of the drug given concomitantly with antibiotic in acute infections. Three double-blind, drug versus placebo studies were conducted in fragile institutionalized or hospitalized patients. Antibiotics were administered simultaneously with RU 41740 in one group and with a placebo in another group. The studies performed by Albarede and Ollivier showed that in acute respiratory infections RU 41740 was well tolerated and resulted in a more rapid improvement of severity score. Grassi and al. studied chronic bronchitis patients admitted for acute on chronic episode. RU 41740 produced a more rapid improvement in the most severely ill patients, and it was well tolerated. It is concluded that RU 41740 can be initiated safely in acute episodes occurring in subjects with recurrent respiratory infections, and that it results in a faster improvement of clinical symptoms. PMID- 2971187 TI - [Immunotolerance of RU 41740]. AB - RU 41740 is an immunomodulator of organic origin to be used preventively against recurrent respiratory infections in both adults and children. The immunological safety of the drug was first studied on NZB-NZW mice which spontaneously develop a disease resembling lupus erythematosus. The survival rate of these mice and the amounts of protein excreted in the urine were not modified by the compound. Clinically, RU 41740 was well tolerated by the adult mice and their offspring. Studies on immunological safety were also conducted in human adults and children. Compared with pre-treatment values, there were no pathological changes in blood count and differential, erythrocyte sedimentation rate, immunoglobulins E, A, M and G levels, apolipoproteins, haptoglobin, haemolytic complement, antinuclear factors, rheumatic serology and anti-red cell antibodies. In addition, the drug was well tolerated by atopic subjects. PMID- 2971189 TI - [Asthmology 1988]. PMID- 2971188 TI - [Contribution of molecular biology in the field of infectious diseases. The challenge]. PMID- 2971190 TI - [First-line treatment of severe myelomas of the young subject by high-dose melphalan]. AB - Six patients (aged 39 to 55 years) with stage III myelomas were treated with IV high dose melphalan (HDM) 140 mg/m2. They had previously received only one or two courses of conventional chemotherapy. The extrahematological tolerance of the procedure was good. The duration of neutropenias (less than 500/mm3) was 14 to 24 days. Complete remission (CR) was obtained in 4 cases but one patient died in CR of invasive aspergillosis. In the other 3 cases, bone marrow was cryopreserved without ex vivo treatment and reinfused after a 2nd course of HDM. These patients remain in apparent CR 12, 20 and 34 months after the diagnosis although one of them had a pure meningeal relapse treated with a second autograft. These results suggest that CR can be obtained with HDM but at the expense of profound myelosuppression. Long term disease control can be obtained but the role of the 2nd course of HDM with autologous bone marrow transplantation warrants further study. PMID- 2971191 TI - [Mycoplasma pneumoniae infections. A multicenter retrospective study of 182 cases]. AB - The spectrum of 182 hospital Mycoplasma pneumoniae infections, collected over a five-year period, is reviewed. The diagnosis was based on complement fixation test. Mycoplasma infection was encountered mainly in young people, mean age 29 years. Eighty-four per cent presented with an atypical pneumonia, 33 per cent had extrapulmonary manifestations mainly neurological and cutaneous. Acute respiratory failure occurred in 32 cases. Only 5 patients died. Macrolides or tetracyclines remain the most effective antibiotics but spontaneous recovery is usual. PMID- 2971192 TI - [Tuberculosis in patients infected with the human immunodeficiency virus. 30 cases]. AB - Between May, 1983 and September, 1987, 8 per cent of the patients hospitalised for an HIV infection (i.e. 30 patients, 20 with an ARC and 10 with AIDS) had tuberculosis. The percentage of patients originating from Central Africa or Haiti was important (23 per cent). Tuberculosis was thoracic (76 per cent) and/or extrathoracic (63 per cent). The main organs involved were the lungs (n = 21), the mediastinal lymph nodes (n = 9), the superficial lymph nodes (n = 9), and the liver (n = 8). The pulmonary infection was often multilobar (n = 14), but without caverns. The tuberculin PPD (purified protein derived) test was positive in 63 per cent of ARC patients and in 30 per cent of AIDS patients. The diagnosis of tuberculosis was confirmed in 27/30 patients by culture of Mycobacterium tuberculosis (n = 23) and/or histology (n = 13), and in the remaining patients by response to a specific treatment. In 3 patients with normal X-ray film of the chest, M. tuberculosis could be recovered by culture of the gastric fluid. Antituberculous treatment was effective, but its optimum duration is to be determined since relapse may occur, even after one year of treatment. The side effects of the treatment were unusually frequent (54 per cent). The occurrence of tuberculosis seemed to aggravate the prognosis of the HIV disease, since 57 per cent of the ARC patients reached the stage of AIDS within 6 months on average. These results are in agreement with the new recommendations of the Centers for disease control which include extrapulmonary tuberculosis in the AIDS criteria. However, in our study, pulmonary tuberculosis had the same detrimental effect and should therefore be included in the AIDS criteria. PMID- 2971193 TI - [Cancer of the thoracic esophagus. Therapeutic strategy]. AB - Out of 100 patients with oesophageal carcinoma, 39 will have the tumour resected, 26 will survive the operation, and only 4 will be alive after 5 years. Although surgical resection is the best treatment for squamous cell carcinoma, new trends in pre-operative chemotherapy should provide better results. A therapeutic regimen based on data from the literature and on personal experience is suggested as neoadjuvant chemotherapy in operable and in nonsurgical tumours. Operability should be re-evaluated after 2 courses of chemotherapy according to response rate. Surgical possibilities and post-operative treatment depend on this response. Surgical procedures and the indications of surgery are described. PMID- 2971194 TI - [Impotence caused by venous leakage. Placement of a prosthetic venous bandage]. AB - A new treatment of impotence by venous leakage is reported. It consists of tying the dorsal and lateral aspects of the cavernous body with a bandage of synthetic material. The bandage, inserted on the bulbo-cavernous muscles, restores a "carter effect" and prevents venous leakage during erection. This technique has been used in 20 patients, with satisfactory results in 10 who were followed up for more than 4 months. PMID- 2971195 TI - [Diagnostic consequences of an unusual connection between the two ductal systems: incomplete pancreas divisum]. PMID- 2971196 TI - [Indirect pathogenicity: a cause of failure of various antibiotic treatments in pediatrics]. PMID- 2971197 TI - ["Azygo-portal disconnection", "direct attack" of esophageal varices? In favor of more adequate terminology]. PMID- 2971198 TI - [Who eats stones loses his iron]. PMID- 2971199 TI - [Apical fibrosis in ankylosing spondylitis. Contribution of the mucociliary clearance]. PMID- 2971200 TI - [Treatment of chronic myeloid leukemia by recombinant alpha interferon and hydroxyurea]. PMID- 2971201 TI - [Abdominal actinomycosis. 1 case]. PMID- 2971202 TI - [Metastatic tumors of the small intestine]. PMID- 2971203 TI - [Osteoporosis and nerve compression]. PMID- 2971204 TI - [A complication of cutaneous necrosis following phlebography]. PMID- 2971205 TI - [Destructive forms of recurrent pulmonary tuberculosis among the rural population]. PMID- 2971207 TI - [Present-day dynamics of the thanatogenesis of tuberculosis]. PMID- 2971206 TI - [Valve of laparoscopy in the diagnosis of tuberculosis of female genital organs and tuberculous mesenteric lymphadenitis]. PMID- 2971208 TI - [Founding and development of phthisiological services in Uzbekistan]. PMID- 2971209 TI - [Recurrence of tuberculosis of the respiratory organs]. PMID- 2971210 TI - [State of the red blood and daily erythropoiesis in patients with tuberculosis of the lungs in the USSR in northern Europe]. PMID- 2971211 TI - [Incidence of tuberculosis of the respiratory organs and the composition of newly diagnosed patients]. PMID- 2971212 TI - [Dispensary care of children with mucoviscidosis in an urban pulmonology center]. PMID- 2971213 TI - [Organization of otorhinolaryngology and bronchology services to tuberculous patients with mental diseases]. PMID- 2971214 TI - [Individual health education of patients with tuberculosis]. PMID- 2971215 TI - Laser treatment of peripheral vascular disease. Implications for nursing care. PMID- 2971216 TI - Preliminary X-ray diffraction studies and biochemical characterization of the antitumor protein mitomalcin indicate close similarity to neocarzinostatin. AB - The antitumor antibiotic protein mitomalcin, from the microorganism Streptomyces malayensis, has been purified to apparent homogeneity and crystallized. The crystals belong to space group P2(1)2(1)2(1) and have the following cell parameters: a = 27.2 A, b = 34.1 A, c = 101.7 A, and alpha = beta = gamma = 90 degrees. These crystal properties are extremely similar to crystals of the antitumor protein neocarzinostatin (11.7 kilodaltons [kDa]) from Streptomyces carzinostaticus in spite of differing pH conditions for crystallizing the two proteins and an apparent difference in molecular weight. Gel electrophoresis shows the molecular weight is similar to that of neocarzinostatin. An amino acid composition analysis of mitomalcin indicates that some differences may exist between the two molecules, but a preliminary amino acid sequence analysis of the first 37 residues found no difference in the N-terminal region of the molecule. PMID- 2971217 TI - Biochemically stimulated release of atrial natriuretic factor from heart-lung preparation of Dahl rats. AB - We investigated the effect of sodium chloride and adrenergic agents on the release of atrial natriuretic factor (ANF) using working heart-lung preparations from Dahl salt-hypertension sensitive (S) and Dahl salt-hypertension resistant (R) rats. High concentrations of NaCl moderately increased ANF release, but this was attributed to small increases in left atrial pressure rather than to a direct effect of NaCl on ANF release; S and R rats responded similarly. Neither isoproterenol (beta 1 + beta 2 agonist) nor clonidine (alpha 2 agonist) had any effect on ANF release in the heart-lung preparation. In contrast, phenylephrine (alpha 1 agonist) stimulated ANF release. This could not be accounted for by change in atrial pressure and appeared to be a direct effect. S and R rats both released ANF in response to phenylephrine, but there was a modest tendency for hypertensive S rats to release more ANF than normotensive R rats, which is consistent with previous data on mechanically induced (atrial stretch) ANF release in these strains. PMID- 2971218 TI - Activation of phospholipases during masculine differentiation of embryonic genitalia. AB - We have investigated whether the androgen-induced masculine differentiation of the sex organs involves an induction of phospholipases. We have measured phosphatidylinositol-specific phospholipase C and phosphatidylcholine-specific phospholipase A2 in the reproductive tract of male and female mouse (CD-I) fetuses at the 18th day of gestation. We report here that (1) the activity of these two enzymes is higher in the male genitalia than in the female genitalia; (2) exogenous testosterone at the 13th to 17th day of pregnancy induces both phospholipase A2 and phospholipase C in the female fetal genitalia; and (3) prenatal administration of cyproterone acetate, an antiandrogen, known to produce feminized males, completely prevents the stimulation of phospholipase A2 and C by testosterone in the female fetuses. In the male fetuses, however, cyproterone acetate inhibits the PLC activity but is unable to alter phospholipase A2 activity. These findings provide evidence that the mechanism by which testosterone organizes the genitalia may involve a modification of phospholipases A2 and C. PMID- 2971219 TI - The role of thromboxane A2 in reperfusion injury. AB - Thromboxane A2 (TXA2) receptor antagonists can limit infarct size in models of coronary occlusion and reperfusion, but it was unknown if these compounds can mitigate reperfusion injury. Anesthetized open chest dogs were subjected to left circumflex coronary (LCX) occlusion for 90 min. Two minutes before reperfusion, the dogs were given iv saline (0.9% NaCl) or the TXA2 antagonist SQ 29,548 (0.2 mg/kg + 0.2 mg/kg/hr). Reperfusion was instituted for 5 hr at which time infarct size was determined. Regional myocardial blood flow was determined before, during, and after occlusion. SQ 29,548 treatment resulted in a significant reduction in infarct size (57 +/- 7 and 34 +/- 8% of the left ventricular area at risk infarcted in the saline and SQ 29,548 groups, respectively). No differences in collateral flow during occlusion were observed between groups, but SQ 29,548 treatment resulted in a significantly higher subendocardial reperfusion flow (54 +/- 10 and 93 +/- 14 ml/min/100g for the saline and SQ 29,548 groups, respectively). Thus, TXA2 seems to play a role in exacerbating reperfusion injury and TXA2 receptor blockade may have potential as a mode of therapy for ischemia reperfusion damage. PMID- 2971220 TI - Overview: mechanism of free energy coupling between ATP hydrolysis and ion transport. PMID- 2971221 TI - NMR studies of complexes of Gd3+ with nucleotides and substrate analogues on sarcoplasmic reticulum Ca2+-ATPase. PMID- 2971222 TI - Molecular characterization of a unique anion pump: the ArsA protein is an arsenite(antimonate)-stimulated ATPase. PMID- 2971223 TI - Ca2+ occlusion in monomeric sarcoplasmic reticulum Ca-ATPase. PMID- 2971224 TI - "Uncoupling" of the sarcoplasmic reticulum Ca2+-ATPase? PMID- 2971225 TI - Inhibition of Ca++-ATPase by limited proteolysis. PMID- 2971226 TI - Binding of calcium to the vanadate complex of FITC-labelled Ca-ATPase. PMID- 2971227 TI - Specificity of the E-P state of the SR Ca-ATPase: implications for the coupling mechanism. PMID- 2971228 TI - Modulation by free ATP of sarcoplasmic reticulum ATPase dephosphorylation. PMID- 2971230 TI - Properties of the catalytic site following thiol modification of the Ca2+-ATPase of sarcoplasmic reticulum from skeletal muscle. PMID- 2971229 TI - Ca2+ uptake and affinity of the Ca2+ pump during single cycle catalysis of the sarcoplasmic reticulum ATPase. PMID- 2971231 TI - The effect of thyroxine on the activity of calmodulin-dependent Ca-ATPase in skeletal muscle sarcolemma. PMID- 2971232 TI - The Ca2+ pump of skeletal muscle sarcoplasmic reticulum. PMID- 2971233 TI - Changes in innominate tilt after manipulation of the sacroiliac joint in patients with low back pain. An experimental study. AB - The purposes of this study were to 1) propose a method to detect sacroiliac joint dysfunction (SIJD), 2) test the interrater reliability of the method on a group of patients with low back pain (LBP), and 3) document changes in innominate tilt after manipulation of the sacroiliac joint. Criteria for SIJD were established by the authors. Twenty-six patients with unilateral LBP were examined independently for presence of SIJD by two examiners. Interrater agreement for presence or absence of SIJD was found to be excellent (Cohen's Kappa = .88). Twenty of the patients who met the criteria for SIJD were randomly assigned to an Experimental Group (n = 10) or a Control Group (n = 10). The left and right innominate bones of these 20 patients were measured for tilt before and after the intervention period. The sacroiliac joint of the patients in the Experimental Group was manipulated during the intervention period, whereas the patients in the Control Group received no treatment. Data were analyzed using a mixed three-factor analysis of variance. The data analysis revealed that the manipulation procedure resulted not only in an altered innominate tilt of the same side but also in an equal and opposite tilt of the opposite side (F = 67.07; df = 1.18; p less than .05). The results indicate that SIJD can be identified reliably in patients with LBP and that a manipulative procedure purported to be specific to the sacroiliac joint changes innominate tilt bilaterally and in opposite directions. PMID- 2971234 TI - Thrombin and plasmin activity in diabetes mellitus and their association with glycaemic control. AB - Abnormalities of haemostasis are common in diabetes mellitus. As indicators of fibrinolysis and coagulation, plasmin and thrombin activity were assessed by assay of the fibrinogen peptide derivatives B beta 15-42 and fibrinopeptide A respectively in 60 diabetic patients and 50 control subjects in a cross-sectional study. Glycosylated haemoglobin (HbA1) correlated with B beta 15-42 (r = -0.26, p less than 0.05) and fibrinopeptide A (r = 0.30, p less than 0.05) in the diabetic patients suggesting that poor glycaemic control (i.e. high HbA1 levels) was associated with depressed plasmin and enhanced thrombin activity. Compared to controls, fibrinopeptide A levels were increased in diabetics (p less than 0.001) irrespective of sex or type of diabetes. B beta 15-42 levels were normal in diabetic females but increased in diabetic men (p less than 0.001) possibly secondary to the activation of coagulation. These results suggest that in diabetes mellitus activation of coagulation is the dominant haemostatic abnormality and that better glycaemic control could influence in-vivo plasmin and thrombin activity favourably. PMID- 2971235 TI - Guide wire angioplasty balloon catheter: preliminary report. AB - A new balloon catheter has been developed for angioplasty. The balloon is more flexible and its walls are thinner than those of conventional catheters. It folds down better yet withstands 8-10 atm of pressure. This advance has been married to new guide wire technology, producing a low-profile angioplasty balloon on a 0.038 inch guide wire. It compared favorably with a standard 7-F angioplasty catheter when tested at 4 atm in a rabbit and has been successfully used to dilate ten peripheral stenoses in four patients. PMID- 2971236 TI - Percutaneous transluminal angioplasty below the knee: new techniques, excellent results. PMID- 2971237 TI - Percutaneous transluminal angioplasty of brachiocephalic vein stenoses in patients with dialysis shunts. PMID- 2971238 TI - Arterial occlusive disease below the knee: treatment with percutaneous transluminal angioplasty performed with low-profile catheters and steerable guide wires. AB - In 98 patients with arterial occlusive disease below the knee who were candidates for limb salvage surgery, percutaneous transluminal angioplasty (PTA) was performed with a low-profile balloon catheter and steerable guide wire system. Sixteen patients had bilateral disease; thus, there were 114 endangered limbs and 145 diseased vessels, including 19 with a single stenosis, 94 with multiple stenoses, and 32 with total occlusions. Primary anatomic success was achieved in all stenosed vessels and in 28 occluded vessels. Initial limb healing without amputation was achieved in 88% of limbs. In four patients occluded vessels could not be recanalized; thus, the four affected limbs were amputated. Two years after PTA, 32 of 37 patients available for follow-up had viable pain-free extremities. Cumulative limb salvage rate at 2 years was 86%. At this time, below-the-knee PTA is still recommended only for limb salvage candidates. However, with this new catheter and guide wire system, success rates have been increased and complication rates decreased. PTA may be useful in selected patients with severe claudication. PMID- 2971239 TI - Percutaneous transluminal angioplasty of infrapopliteal vessels: preliminary results and technical considerations. AB - Infrapopliteal balloon angioplasty was performed in 11 patients facing reconstructive surgery for limb salvage. Twelve separate sessions of angioplasty were performed, during which 15 of 16 diseased tibial runoff vessels were successfully dilated. Three complications occurred during percutaneous transluminal angioplasty (PTA). One was related to the concomitant use of a thrombolytic agent to open a femoropopliteal bypass graft and not related to the angioplasty itself. No limbs were lost as a result of complications. The procedure was a technical success in nine of 12 cases (75%) and an initial clinical success in eight of the nine that were technically successful (89%). Follow-up ranged from 1 to 22 months. Six of the nine cases (67%) remained clinically successful during this period. Preliminary results suggest that infrapopliteal PTA is a safe and effective alternative to surgical reconstruction in selected patients. PMID- 2971240 TI - Percutaneous transluminal angioplasty: delayed response in neurofibromatosis. AB - Percutaneous transluminal angioplasty (PTA) was eventually successful in a patient with renal artery stenosis due to neurofibromatosis. Although the immediate postangioplasty appearance showed little improvement, the residual stenosis has completely resolved at 5 months. Delayed response to PTA has been previously documented in other types of vascular disease. Its occurrence in renal artery stenosis due to neurofibromatosis emphasizes the importance of long-term follow-up and may be a factor in the poor short-term results that some have reported for PTA in patients with this condition. PMID- 2971241 TI - Methods of measuring myocardial blood flow in humans. PMID- 2971243 TI - [Recent advances and trends in cancer chemotherapy using nucleoside analogs]. PMID- 2971242 TI - Effects of coronary bypass surgery and angioplasty on coronary blood flow and flow reserve. PMID- 2971244 TI - [False abdominal surgical emergencies]. PMID- 2971246 TI - [Appendiceal cancer. Clinico-therapeutic aspects]. PMID- 2971245 TI - [Cholestasis due to intestinal parasites migrating into the common bile duct]. PMID- 2971247 TI - [Acute appendicitis in the elderly]. PMID- 2971248 TI - [Predictive factors in biliary calculus surgery]. PMID- 2971249 TI - [Acute pancreatitis. Clinico-therapeutic study]. PMID- 2971250 TI - [The value of the I125-fibrinogen test in the diagnosis of postoperative phlebitis]. PMID- 2971251 TI - [Splenic echinococcosis]. PMID- 2971253 TI - [Diagnostic and therapeutic approaches in neurosurgical inflammatory diseases of ENT origin]. PMID- 2971252 TI - [Splenic cysts]. PMID- 2971254 TI - [Traumatic involvement of the facial nerve in ear surgery]. PMID- 2971255 TI - [Indications and methods for strengthening the tympanic membrane]. PMID- 2971256 TI - [Autologous and allogeneic bone in tympanoplasty]. PMID- 2971257 TI - [Myringoplasty with the temporal fascia. The experience of the Timisoara ENT Clinic over the past 5 years]. PMID- 2971259 TI - [Ambulatory treatment of posttraumatic ruptures of the tympanum by the mentor method]. PMID- 2971258 TI - [Traumatic perforations of the tympanum]. PMID- 2971260 TI - [Agenesis of the stapedius muscle. Morphofunctional aspects]. PMID- 2971261 TI - [Chronic seromucous otitis media in children. The etiopathogenetic, clinical and therapeutic aspects]. PMID- 2971262 TI - [The experience of the Iasi Military Hospital in treating transmission hypoacusis]. PMID- 2971263 TI - [Causes and prevention of transmission hypoacusis in children]. PMID- 2971265 TI - [Rosacea: recent findings on the subject of its etiopathogenesis and therapy]. PMID- 2971264 TI - [Labyrinthine fracture with mixed deafness]. PMID- 2971266 TI - [Ependymoma and degenerative lumbar pathology]. PMID- 2971267 TI - Heterogeneity of glycolytic enzyme activity and isozyme composition of pyruvate kinase in breast cancer. AB - In 6 patients with breast cancer - of whom specimens of the primary tumor as well as one of its metastases were available for examination - we demonstrated intratumoral and intertumoral heterogeneity in expression of activity of the glycolytic enzymes hexokinase, phosphofructokinase, aldolase, enolase and pyruvate kinase. Heterogeneity also existed in isozyme composition of pyruvate kinase. The transition of the tumors towards normal surrounding breast tissue showed either a sharp drop in activity, or a gradual decrease in activity, corresponding to pushing margins or infiltrative growth of the tumor as was demonstrated by histologic examination of these specimens. Likewise, the shift towards expression of K isozyme of pyruvate kinase in breast cancer compared to normal breast tissue could be demonstrated. PMID- 2971268 TI - Second look laparoscopy after treatment of acute salpingitis with doxycycline/benzylpenicillin procaine or trimethoprim-sulfamethoxazole. AB - To have their diagnosis verified, etiology determined and treatment evaluated, 64 patients with a suspected acute salpingitis underwent laparoscopy during which isolates were taken. The patients were then randomized to one of two groups for treatment; doxycycline/benzylpenicillin procaine (DC + BP) or trimethoprim sulfamethoxazole (TMP-SMZ). The results were evaluated three to six months later by second look laparoscopy when adhesions and tubal passage were looked for. Isolates from the cervix were culture positive for Chlamydia trachomatis (CT) in 37 (58%) patients and for Neisseria gonorrhoeae (NG) in 15 (23%). Isolates from the oviducts were positive in 17 (27%) patients of whom 12 had CT and two had NG. Mild salpingitis (Grade I) was found in 16 patients, moderate (Grade II) in 26 and severe (Grade III) in 22 patients. At second look laparoscopy, two patients had totally occluded oviducts, 31 had adhesions but tubal passage on at least one side while 31 patients had healed without any signs of residue. Results at second look laparoscopy showed no statistical difference between the two treatment groups. PMID- 2971270 TI - Treatment of shunt-produced edema. PMID- 2971269 TI - The SCID-hu mouse: murine model for the analysis of human hematolymphoid differentiation and function. AB - The study of human hematopoietic cells and the human immune system is hampered by the lack of a suitable experimental model. Experimental data are presented showing that human fetal liver hematopoietic cells, human fetal thymus, and human fetal lymph node support the differentiation of mature human T cells and B cells after engraftment into mice with genetically determined severe combined immunodeficiency. The resultant SCID-hu mice are found to have a transient wave of human CD4+ and CD8+ T cells and human IgG (immunoglobulin G) in the peripheral circulation. The functional status of the human immune system within this mouse model is not yet known. PMID- 2971271 TI - The utilization of physicians' opinions regarding the optimal frequency of periodical post-graduate courses. PMID- 2971272 TI - [Handicapped children. Hokksund's children's hospital]. PMID- 2971273 TI - D1 dopamine receptor--the search for a function: a critical evaluation of the D1/D2 dopamine receptor classification and its functional implications. AB - The present review focuses on the hypothesized D1/D2 dopamine (DA) receptor classification, originally based on the form of receptor coupling to adenylate cyclase activity. The pharmacological effects of compounds exhibiting putative selective agonist or antagonist profiles at those DA receptors positively coupled to adenylate cyclase activity (D1 DA receptors) are extensively reviewed. Comparisons are made with the effects of putative selective D2 DA receptor agonists and antagonists, and on the basis of this work, the DA receptor classification is critically evaluated. A variety of biochemical, behavioral, and electrophysiological evidence is presented which supports the view that D1 and D2 DA receptors can interact in both an opposing and synergistic fashion. Particular attention is focused on the possibility that D1 receptor stimulation is required to enable the expression of certain D2 receptor-mediated effects, and the functional consequences of this form of interaction are considered. A hypothetical model is presented which considers how both the opposing and enabling forms of interaction between D1 and D2 DA receptors can control behavioral expression. Finally, the clinical relevance of this work is discussed and the potential use of selective D1 receptor agonists and antagonists in the treatment of psychotic states and Parkinson's disease is considered. PMID- 2971274 TI - Comparison of the efficacy of preservative free ipratropium bromide and Atrovent nebuliser solution. AB - The paradoxical bronchoconstriction observed with commercially available isotonic ipratropium bromide nebuliser solution (Atrovent) in patients with asthma results from an adverse reaction to the preservatives, benzalkonium chloride and ethylenediaminetetra-acetic acid (EDTA). The airway response to inhaled Atrovent and preservative free ipratropium bromide nebuliser solutions has been examined in a double blind study. On separate occasions 30 asthmatic subjects inhaled 2 ml of the solutions and airway calibre was measured in terms of FEV1 for 45 minutes. Atrovent nebuliser solution provoked a greater than 20% fall in FEV1 in five of the 30 subjects, whereas this did not occur after preservative free ipratropium bromide. Inhalation of the preservative free solution resulted in more rapid and greater overall bronchodilatation than Atrovent, with mean maximum increases in FEV1 of 29.2% and 18.5% respectively. It is concluded that the risk of paradoxical bronchoconstriction with ipratropium bromide is considerably reduced by removal of benzalkonium chloride and EDTA and that preservative free ipratropium bromide is a more potent bronchodilator than the currently available Atrovent solution. PMID- 2971275 TI - Propranolol inhalation challenge in relation to histamine response in children with asthma. AB - The relation between airway responsiveness to propranolol and histamine was studied in 32 asthmatic children. Propranolol and histamine were given by nebuliser to a maximum dose of 16 mg/ml and 32 mg/ml respectively and the response was measured as the provocative concentration of agonist causing a 20% fall in FEV1 (PC20). A PC20 histamine value of less than 32 mg/ml was obtained in 24 of the 32 children, of whom 15 had a measurable PC20 propranolol (less than 16 mg/ml). In these 24 children the geometric mean PC20 histamine was 4.5 mg/ml and 14.4 mg/ml respectively in those with and without a measurable PC20 propranolol (p = 0.023). There was a linear relationship between histamine and propranolol PC20 values (r = 0.60), and between PC20 histamine and FEV1 % predicted (r = 0.43), but not between PC20 propranolol and FEV1 % predicted (r = 0.38). In an open time course study in 12 children with asthma recovery of FEV1 after inhaled propranolol was incomplete in seven of the children after 90 minutes. When inhaled propranolol was followed by inhaled ipratropium bromide in a further 11 children FEV1 had returned to baseline in all children after 60 minutes. Thus propranolol inhalation can be used in children with asthma to assess the contribution of the beta adrenergic system to the regulation of bronchial smooth muscle tone. The test has several disadvantages in comparison with histamine provocation-long duration, the prolonged action of propranolol, and the fact that only the children with substantial hyperreactivity to histamine react to propranolol. PMID- 2971276 TI - Suppression associated lymphocyte markers in lesions of sarcoidosis. AB - The presence within the epithelioid granulomas of sarcoidosis of lymphocytes of the "helper" (T4 positive) phenotype suggests that, as in granulomatous diseases of known aetiology, active cell mediated immunity plays a part in the disease. It has, however, been noted that T8 positive lymphocytes are also found on occasion in the granulomas. The presence of cells bearing markers associated with suppressor activity in various lesions of sarcoidosis has been investigated with a range of monoclonal antibodies. T4 positive cells were present in all granulomas; T8 positive lymphocytes were present within the epithelioid cell areas in proportions that varied from 5 to 55/100 of T4 cells. Other lymphocyte markers associated with suppression (Leu8 and SN130) were very rare on lymphocytes within granulomas but common on both T4 positive and T8 positive lymphocytes in the perigranulomatous mantle, which suggests that expression of these markers was down regulated in the granuloma or that cells bearing them are unable to migrate into the granuloma. Nevertheless, suppressor cell mechanisms could mediate the spontaneous resolution of most cases of sarcoidosis, and the presence or absence of cells bearing suppression associated phenotypes in the granuloma or its mantle may have prognostic importance. PMID- 2971277 TI - [Cyproterone acetate and cancer of the prostate. Experience in 46 cases]. PMID- 2971278 TI - [Cyproterone acetate. Its value in the treatment of prostatic cancer. Apropos of 45 cases]. PMID- 2971279 TI - Mechanisms of immunologic unresponsiveness induced by ultraviolet-irradiated donor-specific blood transfusions and peritransplant cyclosporine. AB - Recipient pretreatment with UV-B irradiated donor-specific blood transfusions (UV DST) combined with peritransplant cyclosporine on days 0, +1, and +2 leads to permanent cardiac allograft survival in the ACI-to-Lewis rat strain combination. This study investigates the mechanisms of immunologic unresponsiveness induced by UV-DST and CsA by examining several in vitro and in vivo parameters in long-term cardiac allograft recipients. The results of the in vitro studies demonstrate that thoracic duct lymphocytes (TDL) of treated and allografted Lewis rats respond less in a mixed lymphocyte reaction to donor splenic lymphocytes (SpL) by 69%, 75%, and 73% (P less than 0.001) at 30, 50, and 100 days after transplantation, respectively, compared with controls, while the response to a third-party (W/F) SpL is unimpaired. In coculture experiments, the TDL from treated recipients specifically suppressed the response of unmodified Lewis TDL to ACI SpL by 59% and 40% (P less than 0.01) at 30 and 50 days after transplantation, respectively, while responses to W/F SpL were suppressed by only 3-6%. The sera obtained from ungrafted rats transfused with UV-DST suppressed the MLR between unmodified Lewis TDL and ACI SpL by 31% (P less than 0.05) while the sera from UV-DST and CsA-treated and allografted rats specifically suppressed the MLR by 75%, 80% (P less than 0.001) and 37% (P less than 0.01) at 10, 30, and 50 days after transplantation, respectively. In vivo adoptive transfer of 10(4) donor-type dendritic cells (DC) into recipients of beating cardiac allografts at 40 or 60 days after transplantation led to rapid and acute allograft rejection, while the adoptive transfer of 10(8) unseparated SpL obtained at 50 days after transplantation from treated Lewis recipients to syngeneic naive hosts led to a modest but significant prolongation of ACI test cardiac allografts. The transfer of serum from treated and allografted recipients at 10, 30, and 50 days after transplantation led to specific and significant prolongation of test grafts in syngeneic naive hosts. These findings suggest that the mechanisms underlying the in vivo immunologic unresponsiveness induced by pretreatment with UV-DST and peritransplant CsA include inactivation without elimination of class II-antigen presenting cells (APC), generation of specific serum suppressor factor(s) and/or antiidiotypic antibody, and induction of donor-specific suppressor cells. PMID- 2971280 TI - Postirradiation recovery of lymphoid cells in the rat. AB - Whole-body irradiation has been extensively used to ablate immune responsiveness in rodent recipients in adoptive allograft assays. This study was undertaken to determine the relative radioresistance and the tempo of regeneration, following whole-body irradiation, of cells involved in the allograft response. Six distinct cell populations have been identified in the lymphoid tissues of rats subjected to sublethal whole-body irradiation. The relative representation of these subpopulations was significantly different from that in nonirradiated controls. NK cells, macrophages, and plasma cells, which are present in very low numbers in cell suspensions prepared from normal lymphoid tissues, made up a significant proportion of the residual/regenerating population in the tissues of rats recovering from whole-body irradiation. More significantly perhaps, the mature T cell populations showed a significant increase in the T cytotoxic/suppressor to T helper cell ratio. These observations support the suggestion that a number of the cell types within the mixed cell population observed in the rejecting indicator grafts of irradiated recipients in adoptive allograft assays are host derived. The finding that the T cytotoxic/suppressor population is apparently more radioresistant than the T helper population supports a conclusion that graft rejection in irradiated recipients, restored with pure populations of T helper cells, may not be directly mediated by the injected cells but may be the result of collaboration between these and host-derived cytotoxic cell populations. PMID- 2971281 TI - [Social and occupational sequelae after hospitalization for low back pain]. PMID- 2971283 TI - [Skin complaints of persons employed in the fish industry]. PMID- 2971282 TI - [Low back pain requiring hospitalization. Waiting time, sick leave and treatment prior to hospitalization]. PMID- 2971284 TI - [Atrial fibrillation, abnormal P-waves and enlarged left atrium]. PMID- 2971285 TI - [An optical method of evaluating intramural blood flow during laparoscopy]. PMID- 2971286 TI - [Effective ways of controlling the educational process]. PMID- 2971287 TI - [Laparostomy and planned reoperations in the complex treatment of suppurative pancreatitis]. AB - An experience with using laparostomy and planned reoperations in 22 patients with purulent pancreatitis is presented. A method of creation of open controlled drainage cavity is described which allows to make the timely and adequate revision of the pancreato-duodenal zone and other organs of the abdominal cavity in planned relaparotomies. Criteria of choice of the optimal terms of reoperations are proposed. PMID- 2971288 TI - [Surgery of the heart conduction system in Leningrad: experience with 3000 operations]. AB - The authors make an analysis of the experience with 3239 operations for different disturbances of heart rate. Recommendations are given for different kinds of electro-stimulation. PMID- 2971289 TI - Role of the hare (L timidus) in the transmission of the bovine lungworm Dictyocaulus viviparus. PMID- 2971290 TI - [Gamma-aminobutyric acid metabolism in the rat brain after administration of nootropic agents]. AB - N,N-bis-pyrrolidonomethyl-diaza - 18-crown-6 and piracetam, which are structural analogues of gamma-aminobutyric acid (GABA) and exhibit nootropic properties, affected the GABA metabolism, the content of total protein, DNA and RNA in rat brain after long-term administration at pharmacologically active doses of 100 mg/kg and 400 mg/kg, respectively. As compared with control values, content of GABA in rat brain was increased by 40% after administration of N,N-bis pyrrolodonomethyl-diaza-18-crown-6 and by 28%--after treatment with piracetam. At the same time, activity of GABA transaminase was decreased by 49.4% and 39.8%, respectively, while the decarboxylase activity was unaltered. PMID- 2971291 TI - [The activity of nonspecific bone marrow and spleen suppressors in the latent period of tumor growth]. AB - Female A/Sn mice aged 2-3 and 11-14 months (with spontaneous adenocarcinomas of the mammary gland and tumor-free) and C57Bl/6 mice were followed after treatment with DMBA. Suppressor cells were tested for the ability to inhibit in vivo antibody responses to sheep erythrocytes by adoptively syngeneically transferred cells. It was demonstrated that the activity of nonspecific suppressors of the bone marrow and spleen increases considerably in the precancer period. PMID- 2971292 TI - [Experience with the introduction and operation of an automatic information processing system for oncology in the Chuvash ASSR]. PMID- 2971293 TI - [Work experience of the WHO International Reference Center in assessing the effectiveness of self-examination for the early diagnosis of breast cancer (results of 2 years' research)]. AB - A prospective controlled study of the effectiveness of self-examination for early detection of breast cancer has been conducted in Leningrad since 1985. The study was carried out in a cohort of 90,000 females at 28 in- and outpatient clinics within 24 months. As a result, breast cancer was identified in 82 cases. Mean size of primary tumor proved 1.0 cm less and tumor detected 4.8 months earlier than those with standard diagnostic procedures. The cohort is planned to expand to 150,000 by the end of 1988. Data on correlation between application of self examination and breast cancer mortality are expected by 1994. PMID- 2971296 TI - [Cut models of investment material for metal-free ceramics prepared by Zeiser method]. PMID- 2971294 TI - [Economic aspects of the treatment of cervical cancer]. AB - The paper presents data pointing to an 1.5-fold economic gain in the use of rehabilitative treatment for cervical cancer. PMID- 2971295 TI - Preparation of leukocyte-poor platelet concentrates from buffy coats. III. Effect of leukocyte contamination on storage conditions. AB - To study the influence of contaminating leukocytes on the storage conditions of platelet concentrates (PC), various amounts of leukocytes were added to identical PC. From 12 blood donations, 12 leukocyte-poor PC were prepared and pooled. Subsequently, the pool was divided into 12 identical PC. The plasma volume of the PC was 58.6 +/- 0.6 ml, the platelet concentration was 1.01 +/- 0.04 x 10(9)/ml (mean +/- SD) and the red cell contamination did not exceed 10(7) per PC. To 4 groups of 3 PC, pooled leukocytes were added from the same 12 blood donations. The leukocyte contamination for each group of 3 PC was 0.14 +/- 0.05, 1.96 +/- 0.09, 5.53 +/- 0.98 and 13.0 +/- 0.93 x 10(6)/ml (mean +/- SD) for groups I-IV, respectively. The PC were stored for 7 days at 22 degrees C in normal polyvinylchloride bags. A significant correlation was found between increasing concentrations of leukocytes in the PC and the drop in pH (r = -0.93), glucose consumption (r = -0.91), lactic acid production (r = 0.93) and release of lactate dehydrogenase (r = 0.92) during storage of the PC. The excretion of beta thromboglobulin, depletion of platelet adenine nucleotides, decreased ability to incorporate 3H-adenosine into metabolic nucleotides and poor morphology of the platelets were also significantly correlated with an increased number of leukocytes in the PC. These data show that high concentrations of leukocytes in PC have a significant detrimental effect on the viability of platelets during storage at 22 degrees C. We conclude that for good storage conditions of PC, the upper limit of leukocytes per PC should not exceed 10(7). PMID- 2971297 TI - [Orthodontic study model]. PMID- 2971298 TI - [Solution to a problem--SG-attachment]. PMID- 2971299 TI - [Clinical test of Kulzer Silicoater process. 10-year study in 4th year]. PMID- 2971300 TI - Increased suppressor cell activity in cattle persistently infected with bovine virus diarrhoea virus. PMID- 2971301 TI - [Inhibitory mechanism of plasma fibronectin on aggregation of elastase-treated platelet]. PMID- 2971302 TI - Immunohistochemical observations of lysozyme in the Paneth cells of specific pathogen-free and germ-free mice. AB - The localization of lysozyme, which may function as an antibacterial agent, was immunohistochemically studied on the mouse Paneth cell secretory granules showing bipartite substructures (central core and peripheral halo). The lysozyme immunoreactivity was observed in the core, but not in the halo. Even in germ-free mice, Paneth cells have many secretory granules and their cores show lysozyme immunoreactivity. It seems likely that mice Paneth cells possess the ability to produce secretory granules containing lysozyme in disregard of the intestinal bacterial milieu. PMID- 2971303 TI - Angiotensin II and progesterone effects on laser Doppler measures of cochlear blood flow. AB - Anesthetized, ovariectomized rats were given bolus injections of angiotensin II (AII) or saline vehicle while Cochlear Blood Flow (CBF) changes were measured using a laser Doppler flowmeter. Experimental animals were pretreated with replacement progesterone and control animals were given oil vehicle injections in place of progesterone. AII increased blood pressure and CBF. Progesterone pretreatment augmented AII-induced pressor responses, while slightly depressing the cochlear blood flow response. These results suggest that AII may be involved in the local, autoregulatory control of CBF, and in possible steroid-related alterations of susceptibility to ototraumas. PMID- 2971304 TI - Gastric blood flow and acid secretion. AB - Prerequisites for proper measurement of gastric mucosal blood flow (MBF) are different according to whether the blood supply of the stomach is being recorded in experimental animals or in humans. In this review the pros and cons of the methods to measure MBF are analysed systematically. Although it is easier to find a procedure for experimental animals none of the method hitherto published proved to be quantitatively accurate for the determination of MBF changes and simultaneous variations in gastric acid production. It is even more difficult to elaborate a method for human studies which should not cause serious inconvencience for the patient. The most one can achieve is a sufficiently accurate estimate of the MBF and its changes in the course of the study. With the help of such procedures it is possible to clarify the interrelation between MBF and acid secretion using interventions or bioactive substances which influence either the mucosal circulation or parietal cell function or both. PMID- 2971305 TI - The effect of low calcium diet, mithramycin, and dichlorodimethylene bisphosphonate on humoral hypercalcemia of malignancy in nude mice transplanted with the canine adenocarcinoma tumor line (CAC-8). AB - The effect of a low calcium diet, mithramycin, or dichlorodimethylene bisphosphonate were evaluated in nude mice with humoral hypercalcemia of malignancy associated with the transplanted canine adenocarcinoma (CAC-8). Low calcium (0.01%) diet significantly reduced serum calcium levels in hypercalcemic nude mice and reduced urine calcium excretion to control levels. Mithramycin (8 mg/kg) decreased serum calcium concentration and urine calcium excretion to the range of control non-tumor-bearing nude mice at day 5 after a single injection, but there was no change in the number of tartrate-resistant acid phosphatase positive osteoclasts in lumbar vertebrae. Osteoclasts from CAC 8-bearing nude mice after mithramycin administration were decreased in size, had small ruffled borders, and increased relative size of clear zones. Dichlorodimethylene bisphosphonate (Cl2MDP) (45 mg/kg) partially reduced serum calcium concentration of hypercalcemic tumor-bearing nude mice, decreased urine calcium excretion to control levels, and markedly reduced the numbers of tartrate-resistant acid phosphatase-positive osteoclasts in lumbar vertebrae. Osteoclasts from Cl2MDP treated nude mice were smaller and had a reduced frequency of ruffled borders than saline-treated hypercalcemic nude mice. In vitro bone resorption induced by CAC-8 extract was significantly reduced by Cl2MDP and mithramycin. The results of these investigations suggest that the hypercalcemia and hypercalciuria associated with HHM in nude mice with CAC-8 are the combined result of altered calcium homeostasis in the bone, kidney, and intestine. Chemotherapeutic agents that specifically affect only bone or feeding a low calcium diet alone may not completely ameliorate the hypercalcemia of HHM. PMID- 2971306 TI - The influence of sex steroid hormones on plasma calcitonin response to the calcium clamp in normal subjects. AB - The influence of sex steroid hormones on plasma calcitonin levels in healthy subjects was studied in 15 males and 10 premenopausal (PREMF) and 12 postmenopausal females (PMF). A standardized ionized calcium stimulus was achieved by means of the calcium clamp technique, and a sensitive RIA was used to determine immunoreactive CT (iCT) in plasma. Plasma iCT levels increased in response to the calcium clamp with an initial peak at 15 or 30 min. The iCT levels then declined but remained at an elevated level for the rest of the 180 min infusion period in all three groups. In males a positive correlation was found between the serum testosterone levels at 0 min and the estimates for initial iCT response, i.e., the change between 0 and 15 min (r = 0.80, n = 11, p less than 0.01), between 0 and 30 min (r = 0.56, n = 15, p less than 0.05) and between 0 and the maximum value (r = 0.68, n = 15, p less than 0.01). In the two female groups no such correlation was found. Serum 17 beta-estradiol or dehydroepiandrosterone sulfate concentrations were not correlated either to basal iCT levels or to the iCT response to the calcium clamp. PMF had lower levels of 17 beta-estradiol than PREMF (p less than 0.001), while the testosterone levels were similar. The iCT levels and response to the calcium clamp showed no significant difference in the two groups. The results indicate that sex difference in the plasma CT levels may be related to different testosterone levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971307 TI - Embryophysiology of motility. PMID- 2971308 TI - Recurrent postdiskectomy low back pain: MR-surgical correlation. AB - The preoperative MR findings in 11 patients, all of whom had developed recurrent low back pain after surgery for herniated lumbar intervertebral disk, were correlated with the surgical findings to determine possible criteria for distinguishing recurrent disk herniation from postoperative scar (extradural fibrosis). The preoperative MR findings agreed with the surgical findings in seven of eight patients with recurrent disk herniation and in six of nine individuals with extradural fibrosis. The most important parameters in differentiating recurrent herniated disk from extradural scar were the configuration and margination of the extradural mass rather than its signal characteristics. The most reliable MR sign for recurrent herniated disk was the presence of a sharply marginated focal polypoid disk protrusion beyond the posterior margins of the adjacent vertebral bodies shown to best advantage on sagittal T1- and T2-weighted and axial T1-weighted spin-echo MR images. Disk herniations usually maintained isointensity with the intervertebral disk of origin, while extradural fibrosis exhibited variable signal intensity. The preoperative diagnosis of extradural fibrosis on MR was based primarily on its irregular configuration and extension. This study suggests that preoperative differentiation between scar and recurrent herniated disk is possible with MR when morphology and topography are considered in addition to signal intensity. PMID- 2971309 TI - Cardiac function in systemic hypertension before and after reversal of left ventricular hypertrophy. AB - In 3 age- and sex-matched groups of subjects--15 normotensives, 15 hypertensives without left ventricular (LV) hypertrophy and 15 hypertensives with LV hypertrophy--the slopes of the regression line obtained by plotting the individual values of LV fractional shortening against the corresponding values of echocardiographic end-systolic stress were compared. The first 2 groups were studied only in control conditions while the third group was restudied after a 20% reduction in LV mass index induced by a long-term antihypertensive treatment and after a 3-week washout period. A significant relation between fractional shortening and end-systolic stress was found in all instances. The slope of this correlation was higher in normotensives (-0.251) and in hypertensives without LV hypertrophy (-0.232) (both p less than 0.01) than in hypertensives with ventricular hypertrophy (-0.079). In this latter group, the slope increased after the reversal of LV hypertrophy (-0.230, p less than 0.01) and remained unchanged (-0.202) at the end of the washout period. No difference was detectable between the slopes obtained in these patients after reversal of LV hypertrophy, both with the antihypertensive treatment on and off, and those of normotensives and hypertensives without LV hypertrophy. Thus, LV hypertrophy attenuates the influence of changes in afterload on LV function. Reversal of LV hypertrophy restores a fractional shortening end-systolic stress relation quite comparable to that found both in normotensives and in hypertensives before the development of LV hypertrophy. PMID- 2971310 TI - Effects of celiprolol on plasma renin, aldosterone, norepinephrine and epinephrine in primary hypertension. AB - Celiprolol is a newly developed cardioselective beta-blocking agent with mild beta 2-agonist and weak alpha 2-antagonist properties. To evaluate the acute (2.5 hours) and chronic (2 weeks) effects of celiprolol (400 mg once a day) on plasma renin, aldosterone, norepinephrine and epinephrine, 20 patients with mild to moderate primary hypertension were studied in a double-blind placebo-controlled crossover trial. Two and one-half hours after the first dose of both placebo and celiprolol, supine and standing measurements of blood pressure showed a significant reduction, whereas plasma norepinephrine increases were comparable with baseline values. Placebo and celiprolol produced similar changes on supine blood pressure and plasma norepinephrine. In 9 patients celiprolol decreased plasma renin (from a mean +/- standard deviation of 1.09 +/- 0.35 to 0.77 +/- 0.52 ng/ml/hr, p less than 0.05) and aldosterone (from 9.2 +/- 3.7 to 6.7 +/- 3.9 ng/dl, p less than 0.05) acutely both supine and standing, but placebo did not change these parameters. Celiprolol increased pulse rate supine (but not standing) as compared with baseline values. After 2 weeks of celiprolol therapy, blood pressure was decreased both supine and standing compared with placebo in 18 patients (140 +/- 18/88 +/- 8 vs 149 +/- 18/94 +/- 7, 136 +/- 18/91 +/- 6 vs 142 +/- 20/97 +/- 9 mm Hg, respectively, each p less than 0.05), without a change of pulse rate when supine and with a reduction when standing. There were no significant changes of plasma renin, aldosterone, norepinephrine and epinephrine levels during chronic therapy compared with placebo. PMID- 2971311 TI - Role of right and left atrial dimensions for release of atrial natriuretic peptide in left-sided valvular heart disease and idiopathic dilated cardiomyopathy. AB - To evaluate the role of atrial dimensions for release of atrial natriuretic peptide (ANP), right and left atrial dimensions (cross-sections) were determined by 2-dimensional echocardiography in 50 patients with left-sided valvular heart disease or idiopathic dilated cardiomyopathy. All patients underwent right- and left-sided heart catheterization with measurement of central hemodynamics. Plasma samples for ANP were withdrawn from femoral vein (ANPv) and ascending aorta. An estimate of right and left meridional atrial wall stress was derived by multiplying cross-sectional areas with pressures of the respective atria. As expected ANPv was closely related to mean right (r = 0.63; p less than 0.001; n = 50) and left atrial pressures (r = 0.61; p less than 0.001; n = 47). Furthermore, a positive correlation between ANPv and right (r = 0.56; p less than 0.001; n = 48) and left (r = 0.30; p less than 0.05; n = 48) atrial cross-sections was obtained. Finally, an excellent relation was found between ANPv and right (r = 0.73; p less than 0.001; n = 48) as well as left (r = 0.58; p less than 0.001; n = 44) meridional atrial wall stress, indicating that atrial wall stress rather than atrial pressures or dimensions alone determines plasma ANP concentrations. However, for identical right and left meridional atrial wall stress 3- to 4-times higher plasma ANPv levels were obtained in patients with idiopathic dilated cardiomyopathy than in patients with left-sided valvular heart disease. This indicates that release of ANP to the same stimulus may be modulated by the nature of the underlying heart disease. PMID- 2971313 TI - Comparative costs of percutaneous transluminal coronary angioplasty and coronary artery bypass grafting in multivessel coronary artery disease. PMID- 2971312 TI - Determinants of plasma atrial natriuretic factor concentrations in congenital heart disease. AB - Seventeen children with congenital heart disease undergoing elective cardiac catheterization were studied to define potential hemodynamic and echocardiographic determinants of plasma atrial natriuretic factor (ANF) concentrations. ANF concentrations in the right ventricle and femoral vein were measured by radioimmunoassay. Right ventricular levels were significantly higher than femoral venous levels (p less than 0.01), but the correlation between the 2 was high (r = 0.87, p less than 0.001). Stepwise multiple linear regression analysis demonstrated that right ventricular ANF concentrations were predicted by a combination of indexed right atrial area measured by 2-dimensional echocardiography, right atrial pressure, pulmonary resistance and heart rate (multiple r2 = 0.83). Femoral venous ANF concentrations were predicted by indexed right atrial area and systolic pulmonary artery pressure (multiple r2= 0.79). It was concluded that there are significant independent predictors of right ventricular and femoral venous plasma ANF concentrations in children with congenital heart disease. The right atrial area indexed by body surface area was the only significant predictor of both central and peripheral plasma ANF concentrations in the multivariate analysis. PMID- 2971314 TI - Initial experience with coronary angioplasty and aortic valvuloplasty using elective semipercutaneous cardiopulmonary support. PMID- 2971315 TI - Balloon dilation angioplasty of Blalock-Taussig shunts. PMID- 2971317 TI - Reverse end-diastolic flow velocity on umbilical artery velocimetry in high-risk pregnancies: an ominous finding with adverse pregnancy outcome. AB - Systolic/diastolic ratios of umbilical velocimetry have been used to assess downstream placental vascular resistance. Reverse end-diastolic flow velocity during end diastole suggests extreme abnormality in waveform and resistance. We reviewed our experience of patients showing reverse end-diastolic flow velocity over a 2 1/2-year period. Out of 550 high-risk patients studied with umbilical velocimetry over this period, 12 patients showed the finding. All patients were delivered of small for gestational age fetuses and the perinatal mortality in this group was 50%. There was also significant perinatal morbidity as judged by cesarean section performed because of fetal distress, low Apgar scores, days in neonatal intensive care, prematurity, placental infarcts, and lethal anomalies. These findings suggest that reverse end-diastolic velocity on umbilical velocimetry is associated with catastrophic perinatal outcome, and aggressive perinatal management may be advised in this group of patients. PMID- 2971316 TI - The application of uterine and umbilical artery velocimetry to the antenatal supervision of pregnancies complicated by maternal sickle hemoglobinopathies. AB - To assess the efficacy of Doppler flow velocimetry in predicting fetal compromise and neonatal outcome in pregnant women with sickle cell hemoglobinopathies, a prospective study was conducted of 96 patients, 48 with sickle cell hemoglobinopathy (8 with SS and 40 with AS hemoglobin) and 48 low-risk AA hemoglobin controls. All subjects were followed biweekly from the third trimester of pregnancy through delivery with uterine and umbilical artery velocimetry, nonstress, tests, and hematocrit and blood pressure measurements. An abnormal systolic/diastolic ratio was defined as a value greater than or equal to 3. The incidence of abnormal systolic/diastolic ratios for uterine or umbilical arteries was significantly higher in pregnant women with SS hemoglobin (88%) when compared with patients with AS (7%) and AA (4%) hemoglobin. In addition, the abnormal systolic/diastolic ratios for both umbilical and uterine arteries are correlated with abnormal nonstress test results. The nonstress test results became abnormal on average 3 weeks after the systolic/diastolic ratios did. The presence of abnormal systolic/diastolic ratios for umbilical and uterine arteries is predictive of fetal distress and infants small for gestational age. The high incidence of concordant uterine and umbilical artery abnormal systolic/diastolic ratios in pregnant women with SS hemoglobinopathy, which were identified earlier than were abnormal nonstress results, suggests an important parameter in the monitoring of these high-risk pregnancies. PMID- 2971318 TI - Atrial natriuretic peptide production in association with nonimmune fetal hydrops. AB - The presence and elevation of atrial natriuretic peptide in fetuses has not previously been demonstrated. This study of right atrial pacing in fetal lambs demonstrated a threefold to fourfold increase in atrial natriuretic peptide during the production of fetal hydrops. Its rate of return to a normal level paralleled the clearance of fetal hydrops. Its possible role in fetal cardiovascular hemodynamics is discussed. PMID- 2971319 TI - Evidence for a local change in the progesterone/estrogen ratio in human parturition at term. AB - The purpose of this study was to determine if a local change in the concentrations of estrogen and progesterone occurs in the amniotic fluid during human parturition at term. Amniotic fluid was retrieved from 20 women in active labor and from 20 women not in labor. Patients were matched for maternal age and gestational age. Estradiol, estriol, progesterone, and dehydroepiandrosterone sulfate were measured in amniotic fluid by radioimmunoassay. For women in active labor, the median concentrations (range) of these steroid hormones were 2.4 ng/ml (1 to 9.2 ng/ml), 1661.2 ng/ml (556.6 to 3928.1 ng/ml), 12.3 ng/ml (4.7 to 41.4 ng/ml), and 187.5 ng/ml (61.2 to 470 ng/ml), respectively. The median concentrations (range) for women not in labor were 1.6 ng/ml (0.3 to 5.7 ng/ml), 684.2 ng/ml (70.6 to 2103.1 ng/ml), 13.2 ng/ml (7.5 to 63 ng/ml), and 65.6 ng/ml (20 to 334 ng/ml), respectively. Thus spontaneous human parturition at term was associated with significantly increased amniotic fluid concentrations of estradiol, estriol, and dehydroepiandrosterone sulfate (p = 0.02, p = 0.002, p = 0.003, respectively). The progesterone/estrogen ratios (progesterone/estradiol and progesterone/estriol) were significantly lower for women in active labor compared with those not in labor (p = 0.002 and p = 0.0006, respectively). We conclude that a local change in the progesterone/estrogen ratio occurs during human parturition at term. PMID- 2971320 TI - Cardiovascular collapse during laparoscopy: a report of two cases. AB - Cardiovascular collapse is a rare but serious complication of laparoscopy. Presented here are two cases of sudden cardiovascular collapse during laparoscopy. Possible causes include severe atypical vagal reactions and carbon dioxide embolism. Arterial blood gases indicated pulmonary vascular shunting. PMID- 2971321 TI - Thymoxamine reverses phenylephrine-induced mydriasis. AB - We performed a randomized double-masked evaluation of the alpha-adrenergic blocking agent thymoxamine (0.1%) as compared to placebo for the reversal of phenylephrine-induced mydriasis. Topically applied thymoxamine reversed the mydriasis from a single drop of 2.5% phenylephrine in 36 of 40 eyes (90%) within one hour. The mydriasis was completely reversed in 25 of 40 eyes (63%). Eyes with blue irides responded more quickly and more completely than did those with brown irides. The 40 contralateral eyes, which had also been dilated with phenylephrine, remained dilated or dilated further after receiving a placebo eyedrop. Twenty subjects (50%) reported mild transient ocular irritation upon instillation of thymoxamine. Thymoxamine was useful in individuals with narrow anterior chamber angles who were at risk of acute closed-angle glaucoma following dilation with an adrenergic agent. PMID- 2971322 TI - Paraneoplastic night blindness with malignant melanoma. AB - A 69-year-old hyperopic man developed acute night blindness and hallucinations of shimmering lights three years after resection of a cutaneous malignant melanoma. There were no metastatic ocular lesions and he had received no medications. His electroretinogram showed abnormalities comparable to those of patients with congenital stationary night blindness with myopia. Metastatic melanoma was recognized several months later. His electroretinographic responses were also identical to those ascribed to vincristine therapy in a previously described patient with malignant melanoma. Our findings showed that acquired night blindness, apparently resulting from interruption of intraretinal rod signal transmission, can be a paraneoplastic effect of a malignant melanoma. PMID- 2971323 TI - Ouabain induces secretion of proatrial natriuretic factor by rat atrial cardiocytes. AB - Neonatal rat atrial and ventricular cardiocytes in primary culture secrete proatrial natriuretic factor (proANF), the 126-amino acid precursor of atrial natriuretic factor (ANF). The cellular mechanisms of proANF secretion differ in the two cell types: atrial cardiocytes store proANF in abundant secretory granules before secretion, whereas ventricular cardiocytes secrete the precursor rapidly after synthesis. In this study, we used the Na+-K+ adenosinetriphosphatase (ATPase) inhibitor ouabain to investigate the functional significance of these differing secretory mechanisms. Ouabain increased immunoreactive ANF (iANF) secretion by atrial cardiocytes 1.5- to 2.4-fold. Metabolic labeling studies demonstrated that proANF was the form of iANF released in response to ouabain. Ventricular secretion of iANF was unaffected by the Na+ K+-ATPase inhibitor. These observations are consistent with a model in which atrial cardiocytes are able to release proANF via a regulated secretory pathway, whereas ventricular cardiocytes utilize a constitutive secretory pathway. The ability of ouabain to stimulate proANF secretion suggests that an increase in intracellular calcium concentration may promote fusion of secretory granules with atrial cell membranes thereby mediating exocytosis of stored proANF. PMID- 2971324 TI - Perinatal expression of the atrial natriuretic factor gene in rat cardiac tissue. AB - The expression of the atrial natriuretic factor (ANF) gene was examined in the perinatal rat ventricle. The late fetal ventricle (day -3 relative to parturition) demonstrated a modest level of ANF gene expression. This expression increased at the time of birth and peaked on day +1 after birth. Atrial ANF immunoreactivity (irANF) increased gradually during the postpartum period (day +1 to 2-3 wk of age), while ventricular irANF as well as ventricular ANF mRNA levels fell over the same interval. This decrease in ANF gene expression resulted from a decrease in activity of the same major transcription start site employed in the adult atria and ventricle. Plasma irANF levels were very high in the neonatal period and decreased over the ensuing 2-3 wk, a profile similar to that seen for ventricular ANF gene expression. In situ hybridization histochemistry revealed that, while adult rat ventricular ANF mRNA was predominantly confined to the subendocardial layer, neonatal ventricles expressed the ANF gene throughout approximately 50% of the inner wall thickness. These findings suggest that the ANF gene is responding to regulatory stimuli that appear at or near the time of parturition and dissipate in the early neonatal period. PMID- 2971325 TI - Antigen presenting cells in human decidual tissue: III. Role of accessory cells in the activation of suppressor cells. AB - Human decidual antigen presenting cells (DAPCs) exposed to fetal cells in vitro induce generation of suppressor T cells among a population of peripheral blood lymphocytes. Human lymphocyte antigen (HLA)-class II positive antigen presenting cells were isolated from early normal human decidual tissue and from peripheral blood (PAPCs) by adhering Ficoll-Paque separated cell suspensions to fibronectin. In contrast to PAPCs, DAPCs pulsed with fetal antigens induced a radio-sensitive, Leu 1,2-positive T suppressor cell population. A nylon wool adherent B cell population is required during the in vitro induction of the suppressor cells. These suppressor cells impair primary mitogen and mixed lymphocyte culture (MLC) responses, generation of anti-trinitrophenyl (TNP) cytotoxic T lymphocytes, and antibody response of autologous and allogeneic lymphocytes. Only intact viable embryonic cells can effectively confer upon DAPCs the ability to induce T suppressor cells. The T suppressor cell induction by DAPCs primed with fetal antigens is restricted by the major histocompatibility complex. Our results show that the HLA-DR molecules are the most prominent restriction elements. PMID- 2971327 TI - Factitious malignant hyperthermia. PMID- 2971326 TI - [Effect of maternal age, parity and the structure and duration of birth intervals on the incidence of labor complications]. PMID- 2971328 TI - Cardiovascular effects of lorazepam during sufentanil anesthesia. PMID- 2971329 TI - Acute pulmonary thromboembolism treated successfully by balloon angioplasty--a case report. AB - Acute pulmonary thromboembolism frequently occurs in patients on protracted bed rest and by itself can cause acute right ventricular failure. The authors report findings in a patient with this disorder treated successfully by balloon angioplasty. PMID- 2971330 TI - Noninvasive measurement of the human peripheral circulation: relationship between laser Doppler flowmeter and photoplethysmograph signals from the finger. AB - Under certain conditions laser Doppler flowmeter (LDF) signals obtained from the finger pulp may appear very similar to those obtained by use of a direct current (dc) photoplethysmograph (PPG). A combined LDF/PPG system was used in conjunction with a circumference strain gauge as an index of volume change to identify the conditions in which the correlation between these signals was good. Simultaneous LDF and dc PPG measurements were made on 10 normal volunteers by using arterial occlusion and on 7 normal subjects by using the Valsalva maneuver at different elevations of the forearm and hand with respect to the midsternum. By altering the elevation of the upper limb the influence of venous filling on each of the signals during these maneuvers was observed. Since the dc PPG signal always appeared similar to the volume change indicated by the circumference strain gauge, it is concluded that the dc PPG signal is related to blood volume change if allowance is made for the effects of blood oxygenation. In circumstances of low venous filling, however, blood volume changes correlate well with blood flow changes, producing the correlation between the dc PPG and LDF traces. The dc PPG signal may be used as a means of monitoring changes in blood flow in the finger only when venous filling is low and the return remains unrestricted. Thus, in investigations using this method, the relative position of the limb with respect to the heart should always be indicated. The LDF method appears to be a reliable indicator of blood flow changes in the microcirculation irrespective of the degree of venous filling. PMID- 2971331 TI - [Reduction of circulating lymphocytes after giving blood. Effects on the establishment of reference values for CD4 and CD8 lymphocytes]. PMID- 2971332 TI - Method to create and maintain the patency of the bovine mammary papilla. AB - The patency of mammary papillae was reestablished after surgically induced injury. Perforated prosthetic tubes with affixed Dacron tubing or Teflon strips were implanted in 18 abraded papillae of lactating dairy cows and were secured with sutures. Wound healing was assessed by palpation and visual inspection. All wounds, with one exception, healed by first intention. Machine milking, reinstituted on day 5 after surgery, caused no apparent discomfort. Grossly and histopathologically, all implants stimulated a variable degree of mucosal metaplasia and hyperplasia. Only implants with Teflon strips became anchored by fibrotic invasion. Mastitis, tube migration, and milk fistulas were complications of the procedure. PMID- 2971334 TI - V-Y advancement of the inferiorly based rectus abdominis musculocutaneous flap. AB - We describe two cases in which a rectus abdominis musculocutaneous flap was used to reconstruct an ipsilateral groin defect by V to Y advancement. Cadaveric studies undertaken to investigate the likely versatility of this flap showed that it may be advanced to midthigh level. This range can be increased by division of the inguinal ligament and by partial reflection of the rectus abdominis muscle from the skin island. The operative technique and the merits and disadvantages of the flap are discussed. This variation is a useful alternative. It has a considerable excursion potential and may be cosmetically more acceptable than other transposition flaps. PMID- 2971333 TI - Organic treatment for the male sexual offender. PMID- 2971336 TI - Ectopic pregnancy. Surgical intervention and perioperative nursing care. AB - Because of the wide variety of ectopic pregnancies, there is no established routine for surgical removal. Fortunately, many are small, unruptured tubal pregnancies that can be easily removed via laparoscopy. But because these procedures and skills are relatively new, most surgeons continue to use a laparotomy in the treatment of ectopic pregnancy. A skilled OR team is required when faced with a ruptured ectopic pregnancy. The Centers for Disease Control, Atlanta, reports that of the 165 deaths resulting from ectopic pregnancies it studied, hemorrhage was the cause of death in 88% of the cases. According to the study, only one third of these women had surgery, but the researchers suggest that prompt surgical intervention may be the key to preventing mortality. PMID- 2971335 TI - [Peritoneal tuberculosis in west Africa]. PMID- 2971337 TI - Occupational skin diseases, United States. Results from the Bureau of Labor Statistics Annual Survey of Occupational Injuries and Illnesses, 1973 through 1984. AB - The overall incidence rates, numbers, and proportions of occupational skin diseases recorded in the Bureau of Labor Statistics Annual Survey of Occupational Injuries and Illnesses, from 1973 through 1984, were reviewed, and a detailed analysis of occupational skin diseases recorded in the 1984 Annual Survey was performed. Overall incidence rates and numbers of cases declined from 1973 through 1983, but increased slightly in 1984. The major industrial divisions of agriculture and manufacturing have consistently had the highest rates and numbers of cases, respectively; skin diseases have accounted for almost two thirds of all occupational illnesses within agriculture. In the 1984 Annual Survey, 11 industries were ranked in the "Top 15" for both incidence rates and numbers of cases, at the two-digit Standard Industrial Classification level. At the four digit level for manufacturing, four industries were also ranked in the "Top 15" for both indexes. This analysis has identified industries toward which research efforts should be directed to characterize those occupational activities or exposures most responsible for these higher risks. PMID- 2971338 TI - Verrucous malignant melanoma. AB - Five cases of verrucous malignant melanoma--a rare variant of melanoma that was described in 1967, but is rarely mentioned today--are presented and the clinical and histopathologic criteria defining this variant are discussed. The importance of this rare entity in the differential diagnosis of verrucous pigmented skin lesions is emphasized. PMID- 2971339 TI - Occupational disease statistics. In perspective. PMID- 2971340 TI - Interaction between pregnancy zone protein and plasmin. AB - Pregnancy zone protein (PZP, alpha 2-PAG, SP3) was found to bind to plasmin in crossed affino-immunoelectrophoresis using sodium caseinate in the first dimension gel. The plasmin presence in the PZP-plasmin complex was confirmed by addition of antiserum against plasminogen to the gel. In crossed affino immunoelectrophoresis using plasmin in the first dimension gel a non migrative PZP immunoreactive peak appeared, similar to the peak obtained with casein in the first dimension gel. Incubation of mixtures of PZP and plasmin also demonstrated complex formation between PZP and plasmin. The complex between PZP and plasmin could be precipitated not only by anti-PZP antibodies, but also by anti plasminogen antibodies, confirming the interaction between the two molecules. The significance of the binding between plasmin and PZP remains to be elucidated, but it is tempting to speculate that PZP, present on the trophoblastic surface, immobilizes plasmin, rendering this molecule able to perform a local fibrinolytic activity. PMID- 2971341 TI - Plasma and urine beta-thromboglobulin in severe preeclampsia. AB - Beta-thromboglobulin (BTG) has been shown to be a specific platelet protein and can be used as a marker of platelet activation in preeclampsia. Concomitant studies of BTG levels in plasma and urine were performed with eight primiparous severe preeclamptic patients and eight normal primiparous women matched for age. The mean plasma BTG in the severe preeclamptic patients was 186.62 +/- 29.93 ng/ml, and in the control group 45.38 +/- 31.84 ng/ml. The P-value for the difference was highly significant (P = 0.000). In contrast, the mean urine BTG in the study group was 8.42 +/- 4.61 ng/ml, while the mean value for the control group was similar, 5.00 +/- 3.20 ng/ml. The P-value for the difference was not significant (0.05 less than P less than 0.10). These results show that urinary BTG cannot be considered an indicator of platelet activation in severe preeclampsia. A low urinary BTG concentration in the presence of high plasma BTG levels may rather express renal impairment. Failure of BTG renal clearance would contribute to further raising the level of plasma BTG. PMID- 2971342 TI - Biologic markers, genetics, and Alzheimer's disease. PMID- 2971343 TI - Glucosteroid (GS)-dependent immunomodulation of immunoglobulin biosynthesis in vitro. II. Effect of GS on mitogen- and alloantigen-induced Ig synthesis. AB - GS cause an enhancement of T cell regulated Ig synthesis in vitro, while direct B cell response is little affected. The action of GS depends upon the timing of their addition in vitro. Early administration of GS produces stimulation, while late may cause inhibition of Ig synthesis. PMID- 2971344 TI - Glucosteroid (GS)-dependent immunomodulation of immunoglobulin biosynthesis in vitro. III. Effect of GS on helper and suppressor cells' Ig production in vitro. AB - GS inhibit both the function of spontaneously occurring helper and suppressor T cells, but the latter activity appears to be more sensitive. The process of induction of suppressor T cells by Concanavalin A is partly steroid-sensitive. Active ConA-induced suppressors appear to be more resistant to GS. PMID- 2971345 TI - Modulation of oligosaccharide processing in an exocrine secretory glycoprotein of rat parotid cells by beta-adrenoreceptor activation. AB - Such stimulation of rat parotid acinar cells in vitro modulated the rate of processing of N-linked oligosaccharides in a high-molecular weight (220 kdalton) secretory glycoprotein. Conversion of polymannose-type oligosaccharides to complex-type oligosaccharides was evaluated by sensitivity to endoglucosaminidase H and alpha-mannosidase, and with a specific inhibitor of glucosidases I/II. Oligosaccharide maturation in the 220 kdalton glycoprotein required one-third to half less time in cells exposed to the beta-adrenergic agonist isoproterenol than in controls. PMID- 2971346 TI - [T cell subsets (Tc, Th, Ts, Tsi) and IL2 receptor bearing cells in peripheral blood in patients with acute phase of alcoholic hepatitis]. PMID- 2971347 TI - Acne in the adolescent. PMID- 2971348 TI - Simple rapid method of rumen cannulation. PMID- 2971349 TI - Ligand-receptor interactions involved in the stimulation of Swiss 3T3 fibroblasts by insulin-like growth factors and insulin. AB - 1. The binding of 125I-labelled insulin-like growth factor 1 (125I-IGF-1) to Swiss mouse 3T3 fibroblasts was time- and concentration-dependent. Unlabelled IGF 1 had a slightly higher potency than multiplication-stimulating activity (MSA) in inhibiting the binding of 125I-IGF-1, and insulin gave a parallel inhibition curve at 300-1000-fold lower potency. Chemical cross-linking of bound 125I-IGF-1 to its receptors, followed by polyacrylamide-gel electrophoresis under reducing conditions, revealed a major band of Mr 130,000, the labelling of which was inhibited by IGF-1 or high concentrations of insulin. 2. The binding of 125I-IGF 1 was not affected by either co-incubation or preincubation of the cells with a range of heterologous growth factors and mitogens. However, IGF-1 and MSA each induced down-regulation of 125I-IGF-1 binding sites. 3. The maximal stimulations of DNA synthesis induced by IGF-1, MSA and insulin, in the presence of a synergizing mitogen, were similar. The dose-response curve for insulin was not parallel to those for IGF-1 and MSA; in particular, low concentrations of insulin induced a greater stimulation than expected on the basis of its potency in the inhibition or down-regulation of 125I-IGF-1 binding. 4. The preincubation of 125I IGF-1 with Swiss 3T3 cells at 37 degrees C decreased its ability to bind to a second batch of cells. This inactivation did not occur when the preincubation was performed at 4 degrees C or in the presence of cycloheximide. Chemical cross linking revealed that the cells released an IGF-binding protein, giving a complex of Mr about 48,000. 5. It is concluded that type I IGF receptors mediate the stimulation of Swiss 3T3 cells by insulin-like mitogens, but that insulin probably stimulates the cells through insulin receptors. The cells can modulate the amount of ligand binding, both by down-regulation of the receptors and by the secretion of an IGF-binding protein. PMID- 2971350 TI - Characteristics of the Ca2+ pump and Ca2+-ATPase in the plasma membrane of rat myometrium. AB - A plasma membrane-enriched fraction from rat myometrium shows ATP-Mg2+-dependent active calcium uptake which is independent of the presence of oxalate and is abolished by the Ca2+ ionophore A23187. Ca2+ loaded into vesicles via the ATP dependent Ca2+ uptake was released by extravesicular Na+. This showed that the Na+/Ca2+ exchange and the Ca2+ uptake were both occurring in plasma membrane vesicles. In a medium containing KCl, vanadate readily inhibited the Ca2+ uptake (K1/2 5 microM); when sucrose replaced KCl, 400 microM-vanadate was required for half inhibition. Only a slight stimulation of the calcium pump by calmodulin was observed in untreated membrane vesicles. Extraction of endogenous calmodulin from the membranes by EGTA decreased the activity and Ca2+ affinity of the calcium pump; both activity and affinity were fully restored by adding back calmodulin or by limited proteolysis. A monoclonal antibody (JA3) directed against the human erythrocyte Ca2+ pump reacted with the 140 kDa Ca2+-pump protein of the myometrial plasma membrane. The Ca2+-ATPase activity of these membranes is not specific for ATP, and is not inhibited by mercurial agents, whereas Ca2+ uptake has the opposite properties. Ca2+-ATPase activity is also over 100 times that of calcium transport; it appears that the ATPase responsible for transport is largely masked by the presence of another Ca2+-ATPase of unknown function. Measurements of total Ca2+-ATPase activity are, therefore, probably not directly relevant to the question of intracellular Ca2+ control. PMID- 2971351 TI - Biosynthesis of secreted beta-hexosaminidase in Tetrahymena thermophila. A comparison of the wild type with a secretory mutant. AB - The synthesis and secretion of beta-hexosaminidase was studied in wild type and secretion-deficient Tetrahymena thermophila cells by metabolic labelling and immunoprecipitation. beta-Hexosaminidase is synthesized as a Mr 79,000 polypeptide which is within 10 min converted into a Mr 59,000 form. The Mr 59,000 polypeptide is further processed (within 20 min) into at least three major mature forms of Mr 58,000-54,000, which are almost quantitatively secreted into the culture medium within 1-2 h after their synthesis. Both precursor and mature forms contain asparagine-linked oligosaccharide chains which are cleavable by endoglucosaminidase F, but not by endoglucosaminidase H. Neither [32P]orthophosphate nor [35S]sulphate are incorporated into immunoprecipitable precursor and mature beta-hexosaminidases, suggesting the absence of a phosphorylated recognition marker. Biosynthesis and processing of beta hexosaminidase is apparently unaltered in the secretory mutant MS-1; however the processed polypeptides remain cellular bound in the mutant, indicating that the mutation affects a late event in the secretion pathway of lysosomal enzymes. PMID- 2971352 TI - Phorbol ester stimulation of fibronectin-mediated cell adhesion. AB - Stimulation of Chinese hamster ovary cells with 12-O-tetradecanoyl 13-acetate increases the rate of adhesion to fibronectin-coated substratum. The EC50 of the phorbol ester that initiates the change in kinetics of adhesion is approximately 8 nM and is specific to those phorbol esters which activate protein kinase C. When compared to control cells, cells stimulated with active phorbol esters require a significantly lower amount of fibronectin to support their adhesion, and exhibit 50% adhesion inhibition by a log-fold higher concentration of PB1, a monoclonal antibody which specifically blocks fibronectin-mediated adhesion. These results indicate that stimulation of cells with phorbol esters results in a modification of the fibronectin receptor leading to an apparent increase in the interaction of the receptor with fibronectin. PMID- 2971353 TI - Effects of an anti-tumor polysaccharide, schizophyllan, on interferon-gamma and interleukin 2 production by peripheral blood mononuclear cells. AB - We examined the effect of schizophyllan, a neutral glucan isolated from the culture filtrate of Schizophyllum commune Fries, on the production of interferon gamma (IFN-gamma) and interleukin 2 (IL 2) from the mitogen-stimulated human peripheral blood mononuclear cells (PBMC). When the levels of IFN-gamma and IL 2 in the culture medium of phytohemagglutin (PHA)- or concanavalin A (Con A) stimulated PBMC were measured by radioimmunoassay and enzyme-linked immunosorbent assay (ELISA) respectively, a significant increase in the production of both cytokines by schizophyllan was demonstrated. These results suggest that the increased production of IFN-gamma and IL 2 may be responsible for the anti-tumor activity of this glucan. PMID- 2971354 TI - Biochemical and immunological evidence for a calcium pump in chromaffin granules. AB - ATP stimulated the accumulation of 45Ca2+ by chromaffin granule ghosts which contained 5 mM oxalate to trap transported calcium within the lumen. Inasmuch as the ATP-dependent 45Ca2+ transport was resistant to 25 mM ammonium acetate as well as the proton ionophore, carbonylcyanide-m-chlorophenylhydrazone, the chromaffin granule proton translocating ATPase does not provide the energy for this process. Instead, we suggest that chromaffin granules contain a calcium translocating ATPase which catalyzes the 45Ca2+ uptake directly. The observation that chromaffin granules bind to a monoclonal antibody raised against a calcium pump from bovine brain supports this hypothesis. PMID- 2971355 TI - Evidence for the presence of ANP-precursor material in the rat thymus. AB - Acidic extraction of the thymus from two day old rats followed by purification on Sephadex G-50 gelfiltration revealed the presence of atrial natriuretic peptide like material (IR-ANP) by radioimmunoassay. Verification of the obtained immunoreactivity has been achieved by the use of two different types of antisera, i.e. two antisera directed against ANP (99-126), the other antiserum recognizing the N-terminal fragment (11-37) of the precursor ANP (1-126) molecule. In addition, reverse phase high performance liquid chromatography (RP-HPLC) and high performance gel permeation chromatography (HP-GPC) monitored by the three antisera have been employed for analysis of the extracted IR-ANP. In both systems the IR-ANP corresponded to the 15 kDa-ANP molecule (1-126). Furthermore, by using immunohistochemical techniques a distinct localization of the IR-ANP material could be demonstrated. The outer cortical area of the thymus, containing mostly lymphoid cells, showed extensive immunostaining with the three different antisera. The data reported here indicate that the rat thymus is a source of ANP. PMID- 2971356 TI - Regional distribution of immunoreactive brain natriuretic peptide in porcine brain and spinal cord. AB - In order to elucidate the physiological functions of brain natriuretic peptide (BNP), radioimmunoassay for BNP was established and regional distribution of BNP in porcine brain and spinal cord was investigated. The concentration of immunoreactive (ir-) BNP in porcine whole brain was estimated to be 0.63 pmol/g, 13 times higher than that of ir-atrial natriuretic peptide (ANP). Highest concentrations of ir-BNP were found in the medulla-pons, striatum, and spinal cord. Medium concentration was noted in the hypothalamus. This distribution of ir BNP in porcine brain was found to be different from that of ir-ANP simultaneously measured. Furthermore, BNP and BNP-32 were identified as two major forms of ir BNP in porcine brain. PMID- 2971357 TI - Strategies for the in vivo inhibition of endogenous atriopeptin. AB - Pharmacological strategies have been employed to manipulate the response to atriopeptin (AP-28; ANF 99-126) in rats. These approaches include: heparin infusion which blocks the natriuresis-diuresis produced by exogenous AP infusion or by acute volume expansion; and autoimmune rats with endogenous antibody that blocks the natriuresis-diuresis produced by administration of exogenous AP or by acute volume expansion in anesthetized rats. Administration of the pressor analog 1-deamino-arg8-vasopressin (dAVP) to rats produces an elevation in systemic and right atrial blood pressure and markedly increases atriopeptin blood levels. The dAVP also produces a delayed natriuresis-diuresis which is effectively blunted in the autoimmune or heparin treated rats. The above approaches provide tools, in the absence of a specific receptor antagonist, with which to validate the intrinsic involvement of AP in physiological, pharmacological, or pathophysiological events. PMID- 2971358 TI - Antagonism of 5-hydroxytryptamine1A (5-HT1A) receptor-mediated modulation of adenylate cyclase activity by pindolol and propranolol isomers. AB - The interactions of the stereoisomers of pindolol and propranolol with 5 hydroxytryptamine1A (5-HT1A) binding sites and adenylate cyclase activity were examined in rat hippocampus. (-)Pindolol and (-)propranolol displayed high affinity for 5-HT1A binding sites, and their affinities were not affected significantly by the addition of 10(-4) M GTP to the radioligand assay. The selective 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) decreased forskolin-stimulated adenylate cyclase activity. The (-)isomers of pindolol and propranolol did not affect basal or forskolin-stimulated activity but, at a concentration of 10(-5) M, they reversed the 8-OH-DPAT inhibition of the forskolin-stimulated cyclase activity. The (+)isomers were less potent in producing this effect. These data suggest that (-)pindolol and (-)propranolol are potent antagonists at 5-HT1A receptors in rat hippocampus. PMID- 2971360 TI - Hypertrophic cardiomyopathy in infants of diabetic mothers: an update. AB - Infants of diabetic mothers (IDMs) are at known risk for developing a hypertrophic type of cardiomyopathy. The severity of IDM cardiomyopathy can vary from an incidental finding on echocardiography to an infant with severe symptoms of congestive heart failure. The purpose of this article is to review the pathophysiologic mechanisms involved in the development of cardiomyopathy in IDMs and to discuss the diagnostic tests utilized in making the diagnosis (especially echocardiography) and the potential mechanisms that may result in congestive heart failure. This report will conclude with a review of a 2 1/2 year prospective study of diabetic women who had carefully maintained diabetic control during pregnancy. Although the IDMs in this study continued to have mild evidence of generalized hypertrophy when compared with control newborn infants, none developed symptoms of congestive heart failure. These data support the contention that careful diabetic management in pregnancy reduces the severity of hypertrophic cardiomyopathy in IDMs. PMID- 2971359 TI - Cytotoxicity, calmodulin and DNA lesions in cells treated with streptozotocin. AB - Streptozotocin is a nitrosourea-derivative containing a methyl group instead of a haloethyl group. Treatment of human adenocarcinoma cells with streptozotocin induces DNA lesions. The presence of the lesions is visualized during cell lysis in dilute alkali as changes in fragmentation of prelabelled DNA. The increase in DNA fragmentation is paralleled by increased cytotoxicity. Furthermore prolonged duration of treatment reduces the level of DNA fragmentation. The repair of the DNA lesions is prevented by treating the cells with W-7, an inhibitor of calmodulin. PMID- 2971361 TI - Studies on the pathogenesis of aplastic anemia in Thailand: evidence of immune mediated mechanism. AB - The pathogenesis of aplastic anemia in Thailand was studied by using in vitro progenitor cells culture. In 37 patients who had active disease, the numbers of colonies derived from erythroid and granulocyte-macrophage progenitor cells (BFU E and CFU-GM) were markedly decreased both in the blood and bone marrow as compared to normal controls. Co-culture of patients' cells with normal blood cells was performed in order to verify an immunologically mediated mechanism. In 8 of 26 patients, there were very low numbers of colonies both BFU-E and CFU-GM in the blood and bone marrow with significant suppression of colony formation in co-culture. Suppressor cells may have caused the aplasia in these patients. The rest had low colony formation and no suppression in co-culture. These patients may have absent or defective stem cells. None had normal colony formation. Therefore, aplastic anemia in Thailand may result mostly from defects involving the stem cells. Only some patients had cell mediated suppression of hematopoiesis as detected by co-culture. PMID- 2971362 TI - Neuromotor disabilities within the James Bay Cree pediatric population. PMID- 2971363 TI - Children attending the local centre for education of handicapped children in Maniitsoq/Sukkertoppen, Greenland. Medical data and living conditions. A case/control study. PMID- 2971364 TI - The problem of heart disease in Puerto Rico. PMID- 2971365 TI - T-cell subsets in peripheral blood of patients with newly diagnosed and posttreatment Hodgkin's and non-Hodgkin's lymphomas. Analysis by monoclonal antibody staining and flow cytometry. AB - T-cell subsets in the peripheral blood of patients with Hodgkin's disease (HD) and non-Hodgkin's lymphomas (NHLs) were determined using anti T-cell monoclonal antibodies and flow cytometry. Forty HD patients and 30 NHL patients were evaluated; 76 normal blood donors served as controls. Newly diagnosed (untreated) HD and NHL patients had relatively normal values for percentages of total T cells, helper cells and suppressor cells; their helper/suppressor ratios were also normal. The total lymphocyte count was normal for pretreatment HD, but lower than normal for NHL. Following treatment, both HD and NHL patients showed significantly decreased helper/suppressor ratios, caused by a significant decrease in the percentage of helper cells in HD patients and a significant increase in the percentage of suppressor cells in the small number of NHL patients studied. A small number of NHL patients, followed without specific treatment (passive follow-up), had relatively normal values for percentages of helper and suppressor cells and total T-cells. For both groups of patients off treatment, it is concluded that the lower helper/suppressor ratios are due to the prolonged effects of treatment (predominantly irradiation). PMID- 2971366 TI - [AIDS and syphilis: the lessons of history]. PMID- 2971367 TI - [A world without smallpox]. PMID- 2971368 TI - [Schistosomiasis in the Caribbean]. PMID- 2971369 TI - [Evaluation of maternal and child health services]. PMID- 2971370 TI - [WHO endorsement of intrauterine devices]. PMID- 2971371 TI - [60 years' work of the International Social Security Association]. PMID- 2971372 TI - [Prevalence of epilepsy in Medellin, Colombia, 1983]. PMID- 2971373 TI - [Maternal nutrition during pregnancy: study of women from a rural area of Costa Rica]. PMID- 2971374 TI - [Acute respiratory infections in children under 2 years of age in the rural area of Guatemala]. PMID- 2971375 TI - [Efficacy of BCG vaccination evaluated by the case-control method in Buenos Aires, Argentina]. PMID- 2971376 TI - [Construction of a health status index: methodological proposal and application]. PMID- 2971377 TI - [The socioeconomic crisis and the demand for health professionals in Brazil, 1976 1984]. PMID- 2971378 TI - Nuclear location of the p55 subunit of the IL-2 receptor following activation of the HT-2 T helper cell line. AB - After mitogenic or antigenic stimulation, T cells express interleukin-2 receptors (IL-2R). The mechanism and control of signal transduction following binding of IL 2 to IL-2R are poorly understood. Using two rat monoclonal antibodies (5A2 and 7D4) specific for two distinct epitopes of the p55 subunit of mouse IL-2R, we have studied the cellular localization of this molecule by immunocytochemistry during the IL-2-mediated activation of mouse T helper cell clone HT-2. During the activation cycle, nuclear staining for the p55 subunit of the IL-2 receptor was transiently observed. It is suggested that the transient nuclear location of the IL-2R may play a critical role in the control of T-cell activation, proliferation and/or differentiation. PMID- 2971379 TI - Maternal language environment of children with mental retardation. AB - Maternal language directed towards children with mental retardation and children matched either for language ability or chronological age was compared in free play and instruction situations. Results were consistent with the hypothesis that mothers of children with retardation match their verbal behavior to their children's language ability while at the same time adopting a more pervasive teaching role. Specifically, they were more directive, but only in free-play. When explicitly asked to instruct their child, they changed very little in comparison with control mothers, whose directive behavior altered to levels previously adopted in free-play by mothers of children with mental retardation. PMID- 2971380 TI - Mothers' directiveness in their interactions with their children with and without Down syndrome. AB - Preschool-aged children (11 with and 11 without Down syndrome), individually matched for communicative ability, mental age, and demographic variables, were videotaped while interacting with their mothers during 15-minutes of free play in a simulated playroom. Maternal responsiveness and three components of maternal directiveness were examined in the context of the child's interactional behavior. Both groups of mothers used a high frequency of controls, but used them primarily to support and encourage the child's participation in interaction. Although mothers of children with Down syndrome exerted greater control in most of the aspects of directiveness, they were not less responsive. Mothers of children without Down syndrome were more likely to silently watch their children who, in turn, were more likely to initiate topics than were the children with the syndrome. PMID- 2971381 TI - Family resources and stress associated with having a mentally retarded child. AB - Concepts drawn from family stress theory and current empirical information on families of handicapped children were integrated to further understand factors influencing parental adjustment to stress associated with such children living at home. Internal and external family resources and characteristics of the children were examined. Mothers (n = 60) were asked to complete four relevant questionnaires. Results of multiple regression analyses indicated that characteristics of the child and the family's crisis-meeting resources were significant predictors of various forms of stress. Clinical implications of these findings and current research directions were discussed. PMID- 2971382 TI - Bone and renal components in hypercalcemia of malignancy and responses to a single infusion of clodronate. AB - Increased bone resorption (BR) and increased renal tubular reabsorption of calcium (TRCa) may both be involved in the pathogenesis of hypercalcemia of malignancy (HM). We have evaluated the relative importance of these two mechanisms in 33 patients with HM after extracellular volume expansion and after single infusion of clodronate (C12MDP: 500 mg iv over 8 h). The fasting urine Ca/creatinine ratio was taken as an index of BR (BRI). An index of TRCa was calculated (TRCaI) from a nomogram based on the relationship between urine Ca excretion per unit of glomerular filtration rate and plasma Ca (PCa). Mean (+/- SEM) PCa fell from 3.29 +/- 0.07 to 2.69 +/- 0.05 mmol/l three days after C12MDP (n = 33, p less than 0.001), a response similar to that obtained with repeated daily iv injections of 500 to 1000 mg C12MDP. The pathogenesis of hypercalcemia varied according to the type of neoplasm. BRI was the highest in multiple myeloma and breast tumors. TRCaI was markedly increased in squamous-cells lung, bladder, kidney and liver carcinomas, reaching levels observed in primary hyperparathyroidism. TRCaI was normal in most cases of multiple myeloma. Breast tumors appeared to be heterogeneous with respect to TRCaI. The fall in PCa in response to a single infusion of C12MDP was usually most marked in cancer patients with elevated BRI and normal TRCaI. It was very modest in patients with high TRCaI and slightly elevated BRI. In conclusion, this study confirms that stimulation of bone resorption is not the only mechanism of the maintenance of hypercalcemia of malignancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971383 TI - Paedodontics. PMID- 2971384 TI - Special surgery opens. PMID- 2971385 TI - 'AIDS and the dentist'. PMID- 2971386 TI - The pharmacokinetics of medroxyprogesterone acetate following two different loading dose schedules in advanced carcinoma of the breast. AB - A loading dose of MPA employing a regimen of 1,000 mg six hourly for 8 doses can achieve plateau levels above 100 ng ml-1 within the first 36 h of treatment, without any untoward toxicity. This raises the possibility of shortening the time to response for this agent. The additional factor of the time required to achieve adequate serum levels may explain the apparent contradictions between reports correlating response rates and dose and those correlating serum level and response. PMID- 2971387 TI - Matrix heparan sulphate, but not endothelial cell surface heparan sulphate, is degraded by highly metastatic mouse lymphoma cells. PMID- 2971388 TI - Identification of CD4+, 2H4+ (T8 gamma +) suppressor-inducer cells in normal human epidermis and superficial dermis. AB - Immunohistological staining of frozen sections of normal human skin demonstrated the presence of significant numbers of mononuclear cells expressing novel epitopes associated with CD4-positive suppressor-inducer functions. The cells were located around superficial vessels and within the basal layers of the epidermis and hair follicles. The antigen identified by the various antibodies has been shown to be functionally important in the induction of various suppressor cells capable of abrogating B cell responses to pokeweed mitogen. The presence in the skin of cells with possible down-regulatory functions in the immune response may be significant with respect to surveillance against neoplasms and control of appropriate responses to infectious agents. PMID- 2971389 TI - Painful red hands: a side-effect of leukaemia therapy. AB - Five cases of 'painful red hands', following chemotherapy for leukaemia, are reported. All the patients were women who had received treatment with cytosine arabinoside, 6-thioguanine and adriamycin. Several days after courses of chemotherapy, painful erythematous swelling of the palms and soles developed. In all cases the reaction was self-limiting and did not adversely affect their prognosis. Physicians should be aware of this reaction in order to reassure their patients. No treatment is required and the chemotherapy need not be stopped or altered. PMID- 2971390 TI - In vitro protein kinase C phosphorylation sites of placental lipocortin. AB - Human placental lipocortin is a high-affinity substrate for rat brain protein kinase C in vitro with phosphorylation occurring on serine and threonine residues in a ratio of approximately 2 to 1. Comparison of the ability of various N terminal-truncated derivatives of lipocortin to serve as phosphorylation substrates, and direct analysis of the N-terminal peptides cleaved from 32P labeled lipocortin, indicated that threonine-24, serine-27, and serine-28 were the phosphorylation sites. The possibility is discussed that a lysine residue near the carboxy side of the phosphorylation site was involved in lipocortin interaction with the catalytic site of protein kinase C. PMID- 2971391 TI - Catalytic mechanism of biotin carboxylase: steady-state kinetic investigations. AB - Biotin carboxylase was purified from Escherichia coli by a new procedure, and its steady-state kinetic parameters were examined. MgATP and bicarbonate add to the enzyme randomly, followed by addition of biotin. Both bicarbonate and MgATP add in rapid equilibrium. A catalytic base with a pK of 6.6 is observed in V/K profiles. Inactivation studies also revealed a sulfhydryl group in the active site that is essential for catalysis. It is proposed that the acid-base catalysts are necessary for the tautomerization of biotin, which presumably enhances its nucleophilicity toward the carboxyl group donor. A second enzymic group with a pK of 6.6, whose role is unknown, is seen in Vmax profiles. The pH profiles for the biotin carboxylase catalyzed phosphorylation of ADP by carbamoyl phosphate have the same shape as the profiles for the forward reaction, which demonstrates that the enzymic bases assume the same protonation states for catalysis of transphosphorylation in either direction. The lack of reactivity of thionucleotide analogues of ATP when Mg is used as the divalent metal ion suggests that both metal ions required for reaction coordinate to the nucleotide. The second metal ion appears to be absolutely required for reaction and not merely an activator of the reaction. Characterization of a bicabonate-dependent biotin-independent ATPase activity strongly suggests that carboxylation proceeds via a carboxyphosphate intermediate. PMID- 2971392 TI - Synexin enhances the aggregation rate but not the fusion rate of liposomes. AB - The effect of synexin on the calcium-induced fusion of large unilamellar liposomes was studied by using two assays for the mixing of aqueous contents. The results were analyzed in terms of the mass action kinetic model, which describes the overall fusion reaction as a two-step sequence consisting of a second-order process of liposome aggregation followed by a first-order fusion reaction. By using several different lipid compositions and varying the electrolyte composition, it was possible to select the rate-limiting step of the overall fusion process. When aggregation was the rate-limiting step, as in the case of Ca2+-induced fusion of phosphatidylserine (PS), phosphatidate (PA)/phosphatidylethanolamine (PE) (1:3), and PS/PE (1:3) liposomes, synexin increased the overall fusion kinetics by increasing the aggregation rate constant (up to 100-fold). When aggregation was rapid compared to destabilization of apposed membranes, i.e., fusion was rate limiting, synexin either had no effect or reduced the overall fusion kinetics. In one such case involving liposomes composed of PA/PS/PE/phosphatidylcholine (PC) (10:15:65:10), synexin reduced the fusion rate constant by 50%. The effect of calcium-induced synexin polymerization was investigated by preincubation of synexin with calcium prior to addition of liposomes. Prepolymerization by Ca2+ always decreased the activity of synexin such that it was less than the activity of an equal amount of untreated monomers. However, it was found that the activity of synexin monomers polymerized to an average hexameric size was greater than that of one-sixth as many untreated monomers, with respect to the liposome aggregation rate constant. Neither polymers nor monomers increased the fusion rate constant. PMID- 2971393 TI - Chemical cross-linking of Sm and RNP antigenic proteins. AB - Nuclear extracts, competent for in vitro premessenger RNA splicing, were chemically cross-linked with thiol-reversible reagents in order to study the organization of proteins within ribonucleoprotein particles (RNPs) containing uridine-rich small nuclear RNAs (UsnRNPs). The distribution of select UsnRNP antigens within cross-linked complexes was determined by Western blotting of diagonal two-dimensional gels. On the basis of calculations from the molecular weights of cross-linked complexes containing UsnRNP common proteins B', B, and D, it is proposed that each of these proteins was associated with UsnRNP common proteins E and G. In addition, D' is proposed to be positioned close to D. The spatial distribution of UsnRNP common proteins was such that B' and B could not be cross-linked to D. The data also suggested that the 63-kDa U1 snRNP specific protein was cross-linked to other U1-specific proteins, particularly C, but not to the UsnRNP common proteins. We propose that part of the UsnRNP core of common proteins contains at least two asymmetrical copies of B':B:D:D':E:G with stoichiometries of 2:1:1:1:1:1 and 1:2:1:1:1:1. PMID- 2971394 TI - Characterization of a microtubule-stimulated adenosinetriphosphatase activity associated with microtubule gelation-contraction. AB - A microtubule-stimulated ATPase is associated with particles that are responsible for microtubule gelation-contraction in vitro. These particles have been proposed to be slow axonal transport, component a, particulates (SCAPs) [Weisenberg, R. C., Flynn, J. J., Gao, B., Awodi, S., Skee, F., Goodman, S., & Riederer, B. (1987) Science (Washington, D.C.) 238, 1119-1122]. The SCAP ATPase activity is stimulated approximately twofold by microtubules. The microtubule-stimulated ATPase activity correlates with the occurrence of microtubule gelation contraction. Both microtubule-stimulated ATPase activity and microtubule gelation contraction are inhibited by millimolar calcium, 0.3 M KCl plus 2 mM ethylenediaminetetraacetic acid (EDTA), 5 microM vanadate, and millimolar N ethylmaleimide (NEM). Neither the ATPase activity nor microtubule gelation contraction is affected by high magnesium concentrations (up to 8 mM) or by the anti-ATPase drugs ouabain, oligomycin, sodium azide, and erythro-9-(2-hydroxy-3 nonyl)adenine (EHNA). Magnesium is required for both ATPase activity and microtubule gelation-contraction. Microtubule-stimulated hydrolysis of GTP, CTP, ITP, and UTP is less than 50% of ATP hydrolysis, and microtubule gelation contraction is reduced in these nucleotides. On the basis of these results we propose that the microtubule-stimulated ATPase activity associated with SCAPs is a previously undescribed enzyme that is responsible for microtubule gelation contraction in vitro and that is the likely motor for component a of slow axonal transport. PMID- 2971395 TI - Oligosaccharide structure and amino acid sequence of the major glycopeptides of mature human beta-hexosaminidase. AB - Human beta-hexosaminidase (EC 3.2.1.52) is a lysosomal enzyme that hydrolyzes terminal N-acetylhexosamines from GM2 ganglioside, oligosaccharides, and other carbohydrate-containing macromolecules. There are two major forms of hexosaminidase: hexosaminidase A, with the structure alpha(beta a beta b), and hexosaminidase B, 2(beta a beta b). Like other lysosomal proteins, hexosaminidase is targeted to its destination via glycosylation and processing in the rough endoplasmic reticulum and Golgi apparatus. Phosphorylation of specific mannose residues allows binding of the protein to the phosphomannosyl receptor and transfer to the lysosome. In order to define the structure and placement of the oligosaccharides in mature hexosaminidase and thus identify candidate mannose 6 phosphate recipient sites, the major tryptic/chymotryptic glycopeptides from each isozyme were purified by reverse-phase high-performance liquid chromatography. Two major concanavalin A binding glycopeptides, localized to the beta b chain, and one non concanavalin A binding glycopeptide, localized to the beta a chain, were found associated with the beta-subunit in both hexosaminidase A and hexosaminidase B. A single major concanavalin A binding glycopeptide was found to be associated with the alpha subunit of hexosaminidase A. The oligosaccharide structures were determined by nuclear magnetic resonance spectrometry. Two of them, the alpha and one of the beta b glycans, contained a Man3-GlcNAc2 structure, while the remaining one on the beta b chain was composed of a mixture of Man5-7-GlcNAc2 glycans. The unique glycopeptide associated with the beta a chain contained a single GlcNAc residue. Thus, all three mature polypeptides comprising the alpha and beta subunits of hexosaminidase contain carbohydrate, the structures of which have the appearance of being partially degraded in the lysosome. In the alpha chain we found only one possible site for in vivo phosphorylation. In the beta it is unclear if only one or all three of the sites could have contained phosphate. However, mature placental hexosaminidase A and B can be rephosphorylated in vitro. This requires the presence of an oligosaccharide containing an alpha 1,2-linked mannose residue. Only the single Man6-7 (of the Man5-7-GlcNAc2 glycans) containing site on the beta b chain retains this type of residue. Therefore, this site may act as the sole in vitro substrate in both of the mature isozymes for the phosphotransferase. PMID- 2971396 TI - (Iodoacetamido)fluorescein labels a pair of proximal cysteines on the Ca2+-ATPase of sarcoplasmic reticulum. AB - Previous energy transfer studies [Squier, T. C., Bigelow, D. J., de Ancos, J. G., & Inesi, G. (1987) J. Biol. Chem. 262, 4748-4754] have utilized fluorescent iodoacetamide derivatives covalently bound to the Ca2+-ATPase of sarcoplasmic reticulum (SR), using labeling conditions that completely modify the most reactive of the protein's surface sulfhydryls to a final level of 9 nmol/mg of SR protein. Unambiguous interpretation of these results requires localization of these labeling sites with respect to the primary structure of the Ca2+-ATPase. In the present study, we have used the probe 6-(iodoacetamido)fluorescein (IAF) as a marker for these sites. The IAF-labeled Ca2+-ATPase was completely proteolyzed with trypsin, followed by centrifugation to remove (unlabeled) membrane associated portions. The soluble IAF-labeled tryptic peptides were purified by size-exclusion and reverse-phase high-performance liquid chromatography. Two IAF peptides resulted. The major (4.1 nmol of IAF/mg of starting protein) and minor (1.9 nmol/mg) IAF-peptides were sequenced and were identified, respectively, as Ala673-IAF-Cys674-Cys675-Phe676-Ala677+ ++-Arg678 and as Glu668-Ala669-IAF-Cys670 Arg671. A model is proposed to explain the selectivity of IAF for Cys670 and Cys674 of the approximately 14 surface sulfhydryls of the Ca2+-ATPase. The labeling region, Arg667 through Arg678, has been predicted to be alpha-helical; Cys670 and Cys674 would be adjacent in the helix and imbedded in an Arg cluster. The Arg residues would both attract the anionic IAF and enhance sulfhydryl reactivities by lowering their pK values. PMID- 2971397 TI - Photooxidation of cell membranes in the presence of hematoporphyrin derivative: reactivity of phospholipid and cholesterol hydroperoxides with glutathione peroxidase. AB - The susceptibility of photodynamically-generated lipid hydroperoxides to reductive inactivation by glutathione peroxidase (GPX) has been investigated, using hematoporphyrin derivative as a photosensitizing agent and the human erythrocyte ghost as a target membrane. Photoperoxidized ghosts were reactive in a glutathione peroxidase/reductase (GPX/GRD)-coupled assay only after phospholipid hydrolysis by phospholipase A2 (PLA2). However, enzymatically determined lipid hydroperoxide values were consistently approx. 40% lower than iodometrically determined values throughout the course of photooxidation. Moreover, when irradiated ghosts were analyzed iodometrically during PLA2/GSH/GPX treatment, a residual 30-40% of non-reactive lipid hydroperoxide was observed. The possibility that cholesterol product(s) account for the non-reactive lipid hydroperoxide was examined by tracking cholesterol hydroperoxides in [14C]cholesterol-labeled ghosts. The sum of cholesterol hydroperoxides and GPX/GRD-detectable lipid hydroperoxides was found to agree closely with iodometrically determined lipid hydroperoxide throughout the course of irradiation. Thin-layer chromatography of total lipid extracts indicated that cholesterol hydroperoxide was unaffected by PLA2/GSH/GPX treatment, whereas most of the phospholipid peroxides were completely hydrolyzed and the released fatty acid peroxides were reduced to alcohols. It appears, therefore, that the GPX resistant lipid hydroperoxides in photooxidized ghosts were derived primarily from cholesterol. Ascorbate plus Fe3+ produced a burst of free-radical lipid peroxidation in photooxidized, PLA2-treated ghosts. As expected for fatty acid hydroperoxide inactivation, the lipid peroxidation was inhibited by GSH/GPX, but only partially so, suggesting that cholesterol hydroperoxide-derived radicals play a major role in the reaction. PMID- 2971398 TI - Effects of starvation and streptozotocin-induced diabetes on the activity of phosphofructokinase in the epithelial cells of rat colon. AB - The regulation of phosphofructokinase in the colonic mucosa of 48 h-starved and streptozotocin-induced diabetic rats was investigated. The specific activities of phosphofructokinase from colonic mucosa of starved and diabetic rats were found to be diminished compared with normal controls. The enzyme obtained from the colonic mucosa of normal, diabetic and starved rats showed sigmoidal velocity curves with respect to fructose-6-phosphate, with apparent Km values of 0.6, 0.62 and 0.7 mM, respectively. However, the present results indicated that phosphofructokinase from the epithelial cells of rat colon is not regulated in a manner similar to that of the intestinal enzyme, which was shown to be highly regulated. PMID- 2971399 TI - [Does calmodulin participate in the regulation of the Ca-pump of erythrocytes in vivo?]. AB - Using a highly effective chelator of Ca2+ and 45Ca, the concentration of Cai2+ in human and rat erythrocytes was measured both at normal and accelerated Ca2+ influx into the cells. No effect of the calmodulin-dependent reaction inhibitor R24571 was observed. The Ca-ATPase from saponin-treated erythrocytes was characterized by a high affinity for Ca2+ (K 0.5-0.7 microM). This value is 2-3 times as low as that for Ca2+ concentration causing a 50% increase of the Ca ATPase activity in erythrocyte ghosts obtained during hypoosmotic hemolysis. The Ca-ATPase activity in saponin-treated erythrocytes did not change either under the effect of calmodulin or by R24571. It was assumed that calmodulin did not participate in the regulation of the Ca2+-pump operation in erythrocytes in vivo. PMID- 2971400 TI - [Calmodulin-dependent regulation of Ca,Mg-ATPase activity in plasma membranes of the swine myometrium]. AB - Highly purified plasma membrane (PM) preparations of pig myometrium were found to contain 0.91 +/- 0.22 microgram calmodulin per mg of PM protein. Treatment of membranes with 1 mM EGTA in the presence of 0.2 M NaCl causes the diminution of the calmodulin content down to 3% of the original level. The activity of Ca, Mg ATPase is thereby decreased by 40%. Exogenous calmodulin restores the enzyme activity up to 1.94 +/- +/- 0.30 mumol Pi/mg protein/hour. The maximal activation of Ca, Mg-ATPase is observed with 10(-7) M calmodulin. Calmodulin increases the total ATPase activity of myometrium PM without affecting the Mg-ATPase activity. Trifluoroperazine (20 microM) diminishes the activating effect of exogenous calmodulin on Ca, Mg-ATPase. Calmodulin stimulates Ca, Mg-ATPase at low concentrations of Ca2+(10(-8)-10(-6) M) by decreasing Km for Ca2+ from 0.4.10(-6) M to 2.10(-8) M as well as by increasing Vmax--from 0,8 to 1.42 mumol Pl/mg protein/hour. It is supposed that the activating effect of calmodulin on Ca, Mg ATPase is based on electrostatic interactions of Ca2+-free calmodulin with the enzyme. PMID- 2971402 TI - Quantitative study of gliosis in schizophrenia and Huntington's chorea. PMID- 2971401 TI - [Participation of beta-endorphin in the regulation of the membrane chemoreactive properties of the cortical neurons in rats]. AB - The subcutaneous administration of beta-endorphin and its endogenic accumulation in the process of elaboration of adaptation to the short-term stress influences is accompanied by the change of chemoreactive properties of rat sensorimotor cortex neurons expressed in increasing of the share of areactive cells and lowering of the share of cells reacting by the response activational form to acetylcholine and noradrenaline led to them microiontophoretically. The reaction intensity of cells, both activational and inhibitory forms to the led neurotransmitters in both experimental groups is significantly lower than in control. The revealed changes of chemoreactive properties of neurons, arising under the influence of the increase of beta-endorphin amount in brain may be one of the mechanisms of its participation in the processes of learning and memory. PMID- 2971404 TI - Hemodynamic stress in lateral saccular aneurysms. AB - The flow velocities in glass and silastic lateral aneurysm models were quantitatively measured with the non-invasive laser Doppler method. The influences of the elasticity of the wall, the pulse wave and the properties of the perfusion medium on the intra-aneurysmal circulation were investigated. As shown previously, the inflow into the aneurysm arose from the downstream lip and was directed toward the center of the fundus. Backflow to the parent vessel took place along the walls of the fundus. With non-pulsatile perfusion, flow velocities in the center of the standardized aneurysms varied between 0.4 and 2% of the maximum velocity in the parent vessel. With pulsatile perfusion, flow velocities in the center of the fundus ranged between 8 and 13% of the flow velocity in the axis of the parent vessel. Flow velocities in the aneurysms were slower with a polymer suspension with blood-like properties compared to a glycerol/water solution. Flow velocity measurements near the aneurysmal wall allowed the estimation of the shear stresses at critical locations. The maximum shear stresses at the downstream lip of the aneurysm were in the range of the stresses measured at the flow divider of an arterial bifurcation. The present results suggest that in human saccular aneurysms intra-aneurysmal flow and shear stress on the wall are directly related to the pulsatility of perfusion, i.e. the systolic/diastolic pressure difference and that the tendency to spontaneous thrombosis depends on the viscoelastic properties of the blood, namely the hematocrit. PMID- 2971403 TI - Clodronate for osteolytic metastases due to breast cancer. AB - Breast-cancer patients with multiple osteolytic bone metastases were treated with clodronate (Cl2MDP) 1.6 g/day (17 patients) or placebo (17 patients) for 12 months. Bone pain, extension of bone metastases and formation of new osteolytic foci were reduced by Cl2MDP, and development of severe hypercalcaemia was prevented. After withdrawal of treatment, the patients were followed up for at least 12 months. New bone metastases developed in both groups. There were, however, less fractures and less hypercalcaemia in the Cl2MDP than in the placebo group. The survival rate was higher in the Cl2MDP group than in the placebo group. No side-effects were observed in the Cl2MDP group. PMID- 2971405 TI - Flow profiles and wall shear stress distribution at a hemodialysis venous anastomosis: preliminary study. AB - The phasic velocity field in the vicinity of the venous anastomosis in a hemodialysis angioaccess arteriovenous fistula loop graft (AVLG) is investigated employing a laser Doppler anemometer (LDA) system. Detailed LDA velocity profiles are obtained by sectional survey performed in a transparent, elastic flow model which was fabricated to represent the geometry of the AVLG system under physiological pressure and flow waveforms. The geometry of the flow model was based on a silicone rubber cast obtained from an experimental dog model. In the present study, detailed distribution of velocity profiles is obtained. The distribution of wall shear stress in the model is computed from the slope of the local velocity profiles near the wall. The relationship between the results obtained by flow visualization and the LDA measurement is discussed. PMID- 2971406 TI - [The state of lipid peroxidation and enzymes of calcium transport system in sarcoplasmic reticulum of ischemic myocardium]. AB - In 30 experiments on mongrel dog hearts it was shown that 30 min of total ischemia (37 degrees C) followed by accumulation of MDA in the SR membranes and decrease of their Ca2+-uptake, but had no effect on activity Ca2+-ATPase. After 60-120 min ischemia marked a decrease of Ca2+-uptake and activity Ca2+-ATPase took place, MDA content remained at the increased level. The results show that lipid peroxidation take part in the increase of the permeability of SR membranes for Ca2+ and inhibiting of Ca2+-ATPase. PMID- 2971407 TI - Heterogeneity of drug-dependent platelet antigens and their antibodies in quinine and quinidine-induced thrombocytopenia: involvement of glycoproteins Ib, IIb, IIIa, and IX. AB - The molecular nature of platelet receptors for quinine- and quinidine-dependent antiplatelet antibodies (Q.Ab and Qd.Ab) was studied by immunoblotting. One Q.Ab caused quinine-dependent IgG binding to platelet proteins with molecular weights (mol wts) of 174 Kd and 93 Kd and another to only a 93-Kd protein. A third Q.Ab caused binding to 174-, 140-, 93-, and 57-Kd proteins, while a fourth Q.Ab and a Qd.Ab caused IgG binding to 174- and 18-Kd proteins. Using platelets from patients with Glanzmann's thrombasthenia or Bernard Soulier syndrome and purified GPIIIa, these proteins were shown to be GPIb, GPIIb, GPIIIa, GPIX, and an unidentified 57-Kd protein missing in Bernard Soulier syndrome. Binding to the 93 Kd protein was independent of the PIA1 antigen. Absorption of one Q.Ab with Glanzmann's thrombasthenia platelets revealed different populations of antibodies with different specificities within the one patient. Thus Q.Ab and Qd.Ab are heterogeneous and may be directed toward different epitopes on major platelet glycoproteins. PMID- 2971408 TI - T cell subpopulations defined by monoclonal antibodies after HLA-identical sibling marrow transplantation. II. Activated and functional subsets of helper inducer and cytotoxic-suppressor subpopulations defined by two-colour fluorescence flow cytometry. AB - Two-colour fluorescence flow cytometry was utilized to define subsets within the Leu-3+ (T4+) helper-inducer and the Leu-2+ (T8+) cytotoxic-suppressor T cell subpopulations in recipients of unmanipulated HLA-identical sibling bone marrow transplants. The absolute number of activated cytotoxic-suppressor T cells expressing the HLA-DR and OKT10 activation antigens was increased. The proportion (or relative number) of cells bearing each of these activation antigens was also elevated in both the Leu-2+ and the Leu-3+ subpopulations, with up to 60% of Leu 2+ cells and up to 40% of Leu-3+ cells expressing them. Interestingly, absolute numbers of cells expressing two other activation-associated antigens, the interleukin-2 (IL-2) and transferrin receptors, were not increased in either major T cell subpopulation, although the relative number of Leu-3+ cells expressing both these surface structures was increased early post-transplant. Three putative functional subsets were also enumerated: the Leu-3+, Leu-8+ suppressor-inducer subset was depressed in absolute and relative numbers at most time points post-transplant. The Leu-4+, Leu-15+ suppressor-effector subset subpopulation was normal at all time points posttransplant, while the Leu-2+, 9.3+ cytotoxic precursor subset showed low relative and absolute numbers both early and late post-transplant. None of the abnormalities demonstrated in the present study was correlated with presence or absence of graft-versus-host disease. The study further demonstrates the heterogeneity of the abnormalities in the immune system after human marrow transplantation and lays the basis for functional studies involving these cell populations. PMID- 2971409 TI - Engraftment rates related to busulphan and cyclophosphamide dosages for displacement bone marrow transplants in fifty children. AB - The use of busulphan and cyclophosphamide permitted engraftment in 44 of 49 children receiving 'displacement' bone marrow transplants. Three patients who received T-cell-depleted marrow cells from HLA-haploidentical donors failed to engraft and other graft failures were due to inadequate induction dosage. Our standard schedule comprises busulphan 80 mg/m2/day x 4 days (adjusted if necessary to a minimum of 4 mg/kg/day or a maximum of 5 mg/kg/day) followed by cyclophosphamide 2 g/m2/day x 4 days but reduced so as not to exceed 75 mg/kg/day, a maximum dose preferred for patients with full marrows (e.g. those with thalassaemia major). Of 21 recipients of mixed lymphocyte culture (MLC) negative donor marrow cells with full engraftment at 100 days, there were three late rejections. Of patients transplanted with marrow from MLC-positive donors, one had late rejection after cyclosporin A toxicity had necessitated withdrawal of the drug at day + 146 but six other patients, whose cyclosporin A was stopped routinely 1 year, remain well with full grafts. Ten patients died as a result of graft-versus-host disease. We are therefore exploring new approaches to T-cell depletion and storing autologous marrow for use in the event of graft failure. If necessary, a second transplant with busulphan and cyclophosphamide is best performed at 3 months after full recovery of the host. We conclude that elective transplants can be performed successfully in children with normal immune function without the need for irradiation. PMID- 2971410 TI - Skin explant culture as a model for cutaneous graft-versus-host disease in humans. AB - An in vitro skin explant model for graft-versus-host disease (GVHD) in humans has been used to study the role of effector T cells in the histological pathogenesis of GVHD. In 11 of 12 experiments clear GVHD changes of grades II-IV were induced in HLA-mismatched skin explants cultured with allogeneic T cells sensitized by in vitro mixed lymphocyte culture. The role of effector T cells was investigated by comparing results before and after removal of CD3 positive cells, and CD4 positive and CD8 positive T cell-subsets by antibody and complement cytolysis from responder populations. Only total removal of CD3 positive T cells prevented histopathological lesions of GVHD in the skin biopsy specimens. The results also demonstrated that the CD4 positive population caused the greatest degree of GVHD in vitro in skin biopsy specimens and direct infiltration into skin by cells is not required for changes to become evident. These results confirm the early results on animal models and demonstrate the use of the skin explant model as a tool for studying the biology of GVHD in humans. PMID- 2971411 TI - Significance of atrial natriuretic factor in chronic heart failure. AB - Since the discovery by de Bold et al in 1981 of the natriuretic and diuretic properties of atrial extract a vast literature on the physiological, pharmacological and pathological aspects of atrial peptides has accumulated. In this article we review the significance of atrial natriuretic factor, endogenous and exogenously administered, in chronic heart failure. PMID- 2971413 TI - Laparoscopy in endometriosis. PMID- 2971412 TI - Interpreting the spinal X-ray: 2. AB - Part 1 of this article (July 1988, p. 46) detailed the important features of the interpretation of spinal X-rays in the injured patient and part 2 deals with those patients who present with pain or deformity. The type of examination, the way it is carried out, and the way it is interpreted may all differ from the assessment of the patient who has been injured. PMID- 2971414 TI - Early-onset Huntington's chorea. Diagnostic clues. AB - A patient with early-onset Huntington's chorea is described. The diagnosis had been overlooked during previous contacts with psychiatric services because there was no family history, and features typical of the adult disease were absent. Distinctive findings on CT scan and electroencephalography were observed. PMID- 2971415 TI - Down's syndrome with mania. PMID- 2971416 TI - Down's syndrome and eating disorders. A case study. AB - We describe the case of a 33-year-old man with Down's syndrome and severe eating disorders. The clinical symptoms, and their difference from those of anorexia nervosa, are discussed. The patient responded well to a strict behavioural programme after long, in-patient treatment and was still well at a 2-year follow up examination. PMID- 2971417 TI - Thoracic back pain in rowers and butterfly swimmers--costo vertebral subluxation. PMID- 2971418 TI - Intra-arterial hepatic chemotherapy for liver malignancy. PMID- 2971419 TI - Changes in severity of myocardial infarction and three year survival rates after myocardial infarction in Auckland, 1966-7 and 1981-2. AB - Mortality from coronary heart disease has been declining steadily over the past 20 years in Auckland. The possibility that improved survival contributed to this decline was examined in patients who survived four weeks after myocardial infarction, changes in the severity of the disease being controlled for. Patients who had had myocardial infarction in 1966-7 (191 patients) or 1981-2 (203) were compared to determine whether three year survival rates had changed and the severity of the disease altered. With a coronary prognostic index taken as the measure of severity the 1981-2 group had significantly more severe myocardial infarctions than the 1966-7 group, but despite this the three year survival was significantly better (86% in the 1981-2 group v 75% in the 1966-7 group). These findings suggest that improved survival after myocardial infarction contributed to the decline in mortality. PMID- 2971420 TI - Work of a rehabilitation medicine service. AB - The results of the work of the Edinburgh Rehabilitation Medicine Service in one year were evaluated. Over 1400 new patients were seen, of whom 525 were in hospital at the time. Patients most often referred were those who had had cardiovascular accidents, back pain, lower limb amputations, and head injuries and those convalescing after heart surgery. Most patients had several problems. Many were referred so that they could obtain wheelchairs or other appliances, or for assessment of their ability to drive. Only 3% of the referrals were considered inappropriate. Of the inpatients, 455 (87%) returned home after treatment, but 13 (2%) did not respond satisfactorily to rehabilitation. Twelve per cent (18% of the inpatients) were referred from other health boards, suggesting a need for similar facilities elsewhere. PMID- 2971421 TI - The distribution and plasticity of [3H]vasopressin-labelled specific binding sites in the canary brain. AB - [3H]Vasopressin was used to detect and to quantitate specific binding sites with in vitro receptor autoradiography in the canary brain. A discrete regional distribution of [3H]vasopressin-labelled binding sites was observed. A high density of specific binding sites was present in the medial posterior hypothalamic nucleus, the superficial layer of the optic tectum, the area ventralis of Tsai (AVT), the nucleus pretectalis, the habenula, the nucleus of Darkschewitch and the nucleus interstitialis. A low density occurred in the nucleus robustus archistriatalis of female birds. However, after testosterone treatment the density of the sites in this area as well as in the AVT increased 2 fold. [3H]Vasopressin binding was displaced to the same extent by excess vasopressin as well as by vasotocin. Moreover, the [3H]vasopressin-labelled brain regions are innervated by immunoreactive vasotocin fibers and terminals. It seems likely therefore that the labelled sites represent putative vasotocin receptors. PMID- 2971422 TI - Striatal deficiency of L-pyroglutamic acid in Huntington's disease is accompanied by increased plasma levels. AB - L-Pyroglutamic acid (L-PGA), a cyclized glutamate analogue, was measured in plasma, cerebrospinal fluid and brain tissue of patients with Huntington's disease (HD) and controls. In HD, plasma L-PGA was elevated and possibly reflects an increased requirement of cell membranes to be protected against peroxidative damage. L-PGA was decreased in caudate and putamen of HD patients. We suggest that striatal deficiency of L-PGA in HD is a consequence of neuronal loss which characteristically occurs in HD striatum. PMID- 2971423 TI - [Has transluminal coronary angioplasty replaced the bypass?]. PMID- 2971424 TI - A study of the prevention of hot tapwater burns. AB - A 4-year retrospective study of hot tapwater scalds admitted to Mount Vernon Hospital Burns Unit suggested that the three high-risk groups are young children, the elderly and the mentally and physically handicapped. A household survey was conducted of 60 homes inhabited by people over 70 years of age and 60 homes inhabited by families with toddlers to identify what measures could be taken to prevent hot tapwater scalds. The survey showed that over half of the old people did not have adequate bathing aids and that one-third of all the homes visited had a hot tapwater temperature greater than 60 degrees C, which is the temperature recommended by British Building Services Engineers. The survey also revealed that 50 per cent of the immersion heaters and 25 per cent of the gas boiler central heating systems produced an excessively high hot tapwater temperature. The study thus indicated that both the wider provision of bathing aids to the elderly and a nationwide programme to update old inefficient immersion heater and gas boiler central heating systems would reduce the risk of hot tapwater scalds. The installation of a 'thermoscopic' mixing valve, pre-set and locked at 43 degrees C, at bath and shower outlets can totally eliminate the risk of hot tapwater scalds. At present it is not a feasible option to install these valves in every household, but there is a strong case for installing them in hospitals and residential homes for paediatric, geriatric and mentally/physically handicapped patients. PMID- 2971425 TI - Nonspecific corticosteroid therapy in patients receiving intra-arterial chemotherapy for hepatic metastases of colorectal origin. AB - Cholestatic jaundice induced by hepatic intra-arterial 5-fluorodeoxyuridine (FUDR) demonstrated marked improvement in three patients following treatment with oral corticosteroids. Subsequent "prophylactic" use allowed continuation of chemotherapy and improved quality of life. Corticosteroid use in selected patients with FUDR-induced cholestatic jaundice may be beneficial. PMID- 2971427 TI - Endocrine changes with critical illness. AB - Many alterations in hormonal economy occur predictably with critical illnesses, including changes in thyroid hormones, glucocorticoids, mineralocorticoids, and gonadotrophins. These effects are generally nonspecific and relate primarily to the severity of the illness, rather than to the exact nature of the illness. Thus a wide variety of critical illnesses, ranging from severe pneumonia to hepatic or renal failure, all tend to produce a similar pattern of "stress-induced" hormonal alterations. PMID- 2971426 TI - [Augmentation of the expression of c-myc and c-fos oncogenes as a function of the mechanical activity of the isolated adult rat heart]. AB - c-myc and c-fos oncogenes encode nuclear DNA binding proteins, and are involved in both growth regulation and differentiation. Using the molecular hybridization technique and DNA probes complementary to c-myc and c-fos mRNA, we report an increase in c-myc and c-fos expression level in the isolated beating adult rat heart with reference to the arrested isolated heart. This suggests a causal relationship between mechanical activity of the heart and c-myc and c-fos expression. It evidences for the first time a messenger between mechanical factor and adaptational changes in the phenotype which occurs at the beginning of cardiac hypertrophy. PMID- 2971428 TI - Rehabilitation as a means of the integration of disabled persons into society. PMID- 2971429 TI - [The response of plasma beta-endorphin to postoperative pain after epidural morphine]. PMID- 2971430 TI - Platelets in allergy. Assays and interpretation. PMID- 2971431 TI - Alteration of fibronectin receptors (integrins) in phorbol ester-treated human promonocytic leukemia cells. AB - We describe the isolation of human fibronectin receptors (integrins) from two nonadherent promonocytic cell lines and from peripheral blood monocytes. Integrins purified from U-937 and THP-1 cells exhibited identical electrophoretic migrations on sodium dodecyl sulfate gels run under reducing (approximately Mr 150,000) and nonreducing (alpha, Mr 160,000; beta, Mr 130,000) conditions. Treatment of U-937 or THP-1 cells with phorbol esters induced these cells to express different integrins with electrophoretic mobilities (alpha, Mr 140,000; beta, Mr 115,000, nonreduced) identical to those from normal human peripheral blood monocytes. Receptors isolated from uninduced, nonadherent promonocytic leukemia cells (U-937 and THP-1) were distinct from glycoproteins IIb and IIIa and from leukocyte adhesion molecules (p150/95). However, receptors isolated here did react with an antibody known to block cell adhesion to fibronectin. The differences observed in apparent molecular masses of fibronectin receptors from uninduced and induced U-937 or THP-1 cells are removed by treatment of purified integrins with endoglycosidase F or N-glycanase. In summary, the data presented here demonstrate the purification of integrins by fibronectin affinity chromatography from human leukemia cells and normal peripheral blood monocytes. Our results suggest that these receptors differ in immature and mature monocytic cells, and are altered by glycosylation in the course of cellular maturation. PMID- 2971432 TI - Epidermal growth factor in breast cyst fluid: relationship with intracystic cation and androgen conjugate content. AB - In recent years, several studies focused on the biochemical analysis of breast cyst fluid composition. It has been shown that breast cysts lined by apocrine epithelium contain higher levels of potassium and dehydroepiandrosterone-sulphate as compared to cysts lined by flattened cells, and that women with apocrine cysts are more likely to develop breast cancer. In the present study, we measured the intracystic levels of sodium (Na+), potassium (K+), dehydroepiandrosterone sulphate (DHEA-S), and epidermal growth factor (EGF), a factor which could play a role in the autocrine or paracrine control of breast cancer cell growth as recently proposed by some investigators. Breast cyst fluids obtained by fine needle aspiration from 86 women with gross cystic breast disease were assayed. On the basis of the relative intracystic concentrations of Na+ and K+ two main classes of cysts were defined. An arbitrary cut-off value of 3 for the Na+/K+ ratio seemed adequate to separate these two types of cysts. An inverse relationship was found between the Na+/K+ ratio and DHEA-S concentration, median levels of the androgen conjugate being 3615 micrograms/dl in Na+/K+ less than 3 cysts and 480 micrograms/dl in Na+/K+ greater than 3 cysts (P less than 0.001). EGF levels were found to be significantly higher in Na+/K+ less than 3 cysts as compared to Na+/K+ greater than 3 cysts: 103.26 ng/ml versus 57.22 ng/ml, respectively (P less than 0.001). EGF appeared inversely correlated with total protein concentration in the Na+/K+ greater than 3 cysts, while in the Na+/K+ less than 3 cysts high EGF levels were observed independently of total protein content. In addition, a direct correlation was found between EGF and DHEA-S concentrations. On the basis of these results, the hypothesis can be made that EGF, which is measurable in all breast fluids tested and is nearly undetectable in plasma, is actually produced by the epithelium lining the cyst wall, particularly as far as the Na+/K+ less than 3 cysts are concerned. In view of our results this type of cyst, which has been shown to be lined by apocrine epithelium, appears to be characterized by high DHEA-S and EGF levels. It is suggested that the latter finding could provide a clue for understanding the increased risk of subsequent breast cancer in women bearing apocrine cysts. PMID- 2971433 TI - In vitro generation and antitumor activity of adherent lymphokine-activated killer cells from the blood of patients with brain tumors. AB - A procedure for enrichment in recombinant interleukin-2 (rIL2)-activated natural killer (NK) cells was developed and used for in vitro generation of antitumor effector cells from the peripheral blood of 20 patients with central nervous system (CNS) tumors. In comparison to the patients' unseparated mononuclear cells and nonadherent lymphocytes cultured in the presence of 1000 U/ml of rIL2 for up to 3 weeks, interleukin-2-stimulated lymphoid cells, when purified by adherence to plastic, proliferated better (up to 6,720-fold expansion) and achieved up to five times higher levels of antitumor activity against K562 cell targets and NK resistant glioblastoma cell targets. Two-color flow cytometry analysis showed that cultures of cells purified by adherence to plastic which had the best proliferation contained 10% or less of CD3+Leu19- T-lymphocytes, while the unseparated lymphokine-activated killer cell cultures which proliferated poorly contained up to 85% of CD3+Leu19- T-cells. Cultures of adherent lymphocytes which reached the highest antitumor cytotoxicity were enriched in CD3+Leu19+ effectors (60-80%); the proportion of CD3-Leu19+ NK-cells was not greater than 25% in these cultures. Thus, using the technique of 24- or 48-h activation in rIL2 and adherence to plastic, and in contrast to the results obtained with cells from normal donors, it was not possible to enrich in activated NK cells from the blood of patients with CNS tumors. Instead of activated NK cells, a population enriched in non-major histocompatibility complex-restricted cytotoxic T-cells (CD3+Leu19+) was obtained in cultures from most but not all patients. Low NK cell activity and elevated numbers of circulating CD3+Leu11+ cells seen in the blood of these patients, previously treated by surgery/radiation/chemotherapy and maintained on steroids, could be responsible for the preferential adherence and subsequent expansion to plastic of IL2-activated non-major histocompatibility complex restricted cytotoxic T-lymphocytes. PMID- 2971434 TI - Photodynamic effect in an experimental bladder tumor treated with intratumor injection of hematoporphyrin derivative. AB - Photodynamic therapy (PDT) is an experimental treatment modality for malignant tumors. It is based on the principle that a photosensitizer, such as hematoporphyrin derivative (HPD), is retained in higher concentrations in tumors than in surrounding nonmalignant tissues and that photoactivation of the sensitizer can be used to evoke tumor destruction. However, retention of the systemic injection of HPD is not limited to malignant tissues. This lack of specific tumor localization thus reduces the therapeutic ratio of the treatment and causes skin photosensitivity and possible systemic toxicity. Injection of HPD directly into the tumor, on the other hand, has been shown to yield higher levels of the drug in the tumor and lower levels in normal tissues, in comparison with systemic administration. In this study, we examined the photodynamic effect on s.c. implanted mouse bladder tumors subjected to intratumor (i.t.) and i.p. HPD injections. Tumor cell killing, measured by cell survival, was observed in both the it. and i.p. groups and was dependent on fluence and HPD dosage. However, no significant enhancement of cell killing was observed in the i.t. injected tumors, despite the higher porphyrin levels in these tumors. Histological examination of the effect of PDT on the blood vessels indicated that while cell death accompanied severe hemorrhage in the i.p. injected tumors, in the i.t. tumors there was much less hemorrhage and intact blood vessels remained. This observation suggests that with i.t. administration, direct photodynamic action may play a significant role in the tumor cell killing, in contrast to systemic administration, in which destruction of the blood vessels is believed to be the main cause of tumor destruction. PMID- 2971435 TI - Ultrastructure of myeloma cells in patients with common acute lymphoblastic leukemia antigen (CALLA)-positive myeloma. AB - We investigated the ultrastructure of myeloma cells obtained from four cases of common acute lymphoblastic leukemia antigen (CALLA)-positive myeloma. Clinically, the disease was aggressive and our patients died with a median survival after diagnosis of only 62 days. By light microscopic criteria of Greipp et al., their disease was classified as plasmablastic, immature (two cases), and intermediate. In contrast, the myeloma cells of all four cases were judged to be immature and abnormal on the basis of the electron microscopic observation. Characteristic features were sparse heterochromatin, high to moderate nucleocytoplasmic ratio, nuclear bodies, thin and short rough endoplasmic reticula, scattered pattern of mitochondria, and polysomes consisting of five to six ribosomes, along with irregular nuclear membrane, poorly developed organella, and abnormalities in cytoplasmic structures such as dense bodies, vacuoli, buddings, single-sac loop like structures, multilamellar bodies, and abnormal inclusion bodies. While overlapping each other, it is suggested that the CALLA-positive and the plasmablastic myelomas should be classified separately. Thus, the electron microscopic study, like the immunological marker analysis, provides a useful means for better assessment regarding immaturity and abnormality of myeloma cells. PMID- 2971436 TI - Isolation and characterization of the 36-kDa D-mannose 6-phosphate receptor from porcine testis. AB - A D-mannose 6-phosphate receptor was isolated from total membranes of porcine testis, and its interaction with ligands was examined. The receptor was a glycoprotein comprised of several 36-kDa sub-units with an isoelectric point (pI) of 6.1. The binding of the receptor to the insoluble phosphomannan core occurred in the absence of divalent cations, but was selectively stimulated by MnCl2 and effectively inhibited by D-mannose 6-phosphate, D-fructose 1-phosphate, and pentamannosyl monophosphate. The phosphate group and HO-2 of D-mannose 6 phosphate are important in the receptor-ligand interaction, HO-4 probably contributes to a lesser extent, and HO-1 seems to have no interaction. PMID- 2971438 TI - Medical-social intervention in a 70-year-old Swedish population. A general presentation of methodological experience. AB - The possibilities of improving physical, mental and social functioning or retarding the development of handicap and reducing the need for medical and social services were investigated in an interdisciplinary intervention study in a representative population sample of 1206 70-year-olds. The assessments concerned cognitive function and mental health, earlier and current social interaction and living conditions, state of health including dental status, muscular strength and performance, pulmonary and cardiovascular function, bone mineral content, ADL and dietary habits. The intervention (response rate 76.8%) concerned social, psychological, environmental and medical aspects in 400 and medical aspects only in 406 probands. 400 register controls were sampled for comparison. The intervention period was 2 years, and a follow-up study will be performed at the age of 75. The interventions were guided by the life-style and the activity hypotheses within the resource perspective to promote everyday activities. The study allows comparisons with two previous 70-year-old cohorts investigated with similar methods in Gothenburg. PMID- 2971437 TI - Preparation and application of a pentamannosyl monophosphate-bovine serum albumin conjugate. AB - Pentamannosyl monophosphate, derived from Hansenula holstii O-phosphomannan, was conjugated to bovine serum albumin by reductive amination. The conjugate inhibited the binding of the porcine testis mannose 6-phosphate receptor to the insoluble phosphomannan core. A mannose 6-phosphate receptor with a molecular weight of 200,000 was purified from porcine liver membranes, using an affinity matrix of the conjugate attached to Sepharose 4B. Rabbits were immunised with the conjugate, and the antisera were purified on a phosphomannan core-Sepharose 4B column in order to give an antibody which was specific for the 6-phosphate group and the equatorial HO-4 of D-mannose 6-phosphate. On Western blot analysis using the purified antibodies, ovalbumin, which contained a typical high-mannose type of oligosaccharide, was not recognised. However, a testicular glycoprotein fraction formed an immunostaining band. These results indicate the effectiveness of the conjugate as a ligand for mannose 6-phosphate receptors. The antibodies highly specific for mannose 6-phosphate may be used to detect or purify lysosomal enzymes. PMID- 2971440 TI - Thrombolysis in acute myocardial infarction. AB - This is an extensive review of thrombolysis in myocardial infarction with discussions of the pathophysiology of occlusion, clinical occlusion and rationale for thrombolysis, thrombolytic agents, results of thrombolysis (incidence of reperfusion, left ventricular function, and mortality), complications of thrombolytic therapy, reocclusion, and current recommendations. PMID- 2971439 TI - [Circadian rhythm of atrial natriuretic peptide in healthy man]. PMID- 2971441 TI - Percutaneous transluminal coronary angioplasty during acute myocardial infarction. AB - Intravenous thrombolytic therapy with streptokinase in the setting of acute MI has been shown to be effective in improving left ventricular function, limiting infarct size, and improving early mortality. The benefit of this therapy is greatest when administered within 3 hours and is of minimal benefit when given more than 6 hours from symptom onset. Newer second generation thrombolytic agents such as intravenous r-TPA have been shown to be more effective at establishing patency of acutely thrombosed coronary arteries. TPA treatment produces patency rates similar to those observed with intracoronary administration of streptokinase (65 to 75 per cent). This agent will probably become standard therapy for patients with acute MI. Unfortunately, there are significant problems with systemic thrombolytic therapy. The potential for bleeding complications contraindicates the use of this therapy in patients with recent cerebrovascular events, recent surgery, or other possible bleeding problems. Acute angioplasty of the infarct-related artery has been shown to be effective in restoring blood flow in 85 per cent of patients with acute MI. Preliminary studies have suggested that this therapy, when administered within 4 hours from symptom onset, improves global and regional left ventricular function to a greater degree than intracoronary streptokinase. Patients receiving acute PTCA as a primary reperfusion modality have a lower incidence of post-infarction angina and provokable ischemia by exercise testing. If facilities and skilled personnel are available to perform PTCA within 4 hours from symptom onset, this therapy remains an alternative revascularization modality in patients with acute infarction and contraindications to systemic thrombolytic therapy. However, the benefit of PTCA with regard to reduction in mortality when used in this manner is unproven. PTCA can also be used as an adjunctive therapy administered at some time following systemic thrombolytic therapy. Performing PTCA acutely offers the potential to restore blood flow in 90 per cent of the patients that initially fail thrombolytic therapy. However, despite the use of PTCA in this subgroup, benefits with regard to improved ventricular function and decreased mortality have yet to be conclusively demonstrated. Performing acute PTCA following systemic thrombolytic therapy also incurs a high incidence of bleeding complications. If initial thrombolytic therapy reestablishes vessel patency, similar improvements in ventricular function can be expected even if PTCA is deferred until clinically indicated by evidence of recurrent ischemia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2971442 TI - Selective thrombolysis with low-dose urokinase in chronic arteriosclerotic obstructions. AB - Twenty-one patients, 15 males and 6 females aged 52-75 years, with angiographically demonstrated occlusions of the superficial femoral or popliteal arteries, were treated by low-dose urokinase intraarterial infusion. The obstructions were 2-12 months old and from 7 to 18 cm in length. Urokinase was infused at 50,000 U/h; heparin was simultaneously administered by intravenous route in doses of 800 U/h. The average duration of treatment was 18 h. Effective clot lysis was accomplished in 18 cases (85%); 15 patients had underlying stenoses treated by balloon dilatation to prevent rethrombosis. Of the primarily recanalized arteries, two reoccluted within 4 weeks. PMID- 2971443 TI - Percutaneous transluminal angioplasty of the superficial femoral artery by retrograde catheterization via the popliteal artery. AB - We report the results of 50 angioplasty procedures via the popliteal artery. A 3 year follow-up including control of blood pressures at ankle and toe levels show results comparable to reports in the literature. This new approach for angioplasty of the superficial femoral artery and eventually of coexisting iliac lesions enables treatment of previously inaccessible lesions. The technique is especially suited for lesions close to the takeoff of the superficial femoral artery. PMID- 2971444 TI - Percutaneous transluminal coronary angioplasty in dextrocardia: case report. AB - Percutaneous transluminal coronary angioplasty (PTCA) is described in a patient with dextrocardia. Selective cannulation of the right anterior descending coronary artery (morphologic left anterior descending coronary artery) was achieved with modification of standard guiding catheters "preformed" for the normal, levo-position heart. This report demonstrates the feasibility of PTCA in patients with cardiac malposition who survive into adulthood and develop coronary atherosclerosis. PMID- 2971445 TI - Balloon dilatation of pulmonary artery banding in dogs: an experimental study. AB - The technique at our institution for banding of the pulmonary artery (PA) makes debanding by balloon dilatation theoretically possible. Previously we tested this hypothesis in an in vitro study (see previous article in this issue). This study describes balloon dilatation in dogs. Eight Labrador Retriever dogs had a PA band placed early in life, and when their body weight had doubled, they were restudied and debanded by balloon dilatation or surgical removal. One dog died of a virus infection before debanding and was excluded from the study. Five of the dogs (groups 1 and 2) underwent balloon dilatation with satisfactory relief of the gradient across the PA band. Two dogs (group 3) underwent surgical debanding without a plastic procedure on the PA, and this also produced satisfactory relief of the gradient. All dogs in group 2 and 1 dog in group 3 were studied 3 months later, and 1 dog in group 2 was also studied 6 months after the debanding procedure. The systolic pressure gradient fell from a mean of 54 to 18 mm Hg after the debanding procedure. There was no difference in the gradient after debanding in the balloon group compared to the surgical group. PMID- 2971446 TI - In vitro balloon dilatation of the banded pulmonary artery. AB - The pulmonary artery of unfixated human heart-lung specimens was banded by placing a Dacron tape around the artery and securing the tape with a 5.0 Prolene suture at selected circumferences. The banding was successfully dilated with a balloon catheter in 24 instances. The mean pressure necessary to burst the suture securing the tape was 2.4 atmospheres. PMID- 2971447 TI - Laser angioplasty with a contact probe for the treatment of peripheral vascular disease. AB - Percutaneous transluminal laser angioplasty was carried out for the dilatation of 30 completely occluded peripheral arterial segments and three subtotal stenoses. A neodymium-YAG laser and an optical fibre delivery system with a sapphire tip were used. All three arterial stenoses and 26 of the 30 occluded arterial segments were successfully dilated. In 20 patients additional dilatation was carried out with a balloon catheter. Laser angioplasty failed to recanalise four occlusions, and vessel leakage without clinical consequences occurred in one patient. Reocclusion occurred within 48 h in two patients and after five months in one patient during a follow up period of at least six months. It is concluded that percutaneous transluminal laser angioplasty using direct contact with a sapphire tip is highly effective (89% success rate) in reopening peripheral vascular occlusions. The procedure is safe, and reocclusion of vessels is rare during the six month follow up period. PMID- 2971448 TI - [Radionuclide study of patients with ischemic heart disease after percutaneous transluminal coronary angioplasty]. PMID- 2971449 TI - [The identity of the handicapped child and his parents]. PMID- 2971450 TI - Diversity in the lipocortin/calpactin family. PMID- 2971451 TI - Molecular cloning and expression of the human interferon-gamma receptor. AB - A cDNA encoding the human interferon-gamma receptor was isolated from a lambda gt11 expression library using a polyclonal antireceptor antiserum. The gene for this receptor was identified in a cosmid library and transfected into mouse cells. The human interferon-gamma receptor expressed in mouse cells displayed the same binding properties as in human cells. However, transfected cells were not sensitive to human IFN-gamma, suggesting the need for species-specific cofactors in receptor function. As inferred from the cDNA sequence, the human interferon gamma receptor shows no similarities to known proteins and represents a novel transmembrane receptor. It is most likely the product of a single mRNA and a gene located on chromosome 6q. PMID- 2971452 TI - Selective recycling of the mannose 6-phosphate/IGF-II receptor to the trans Golgi network in vitro. AB - Mannose 6-phosphate receptors carry soluble lysosomal enzymes from the trans Golgi network (TGN) to prelysosomes, and then return to the TGN for another round of lysosomal enzyme sorting. We describe here a complementation scheme that detects the vesicular transport of the 300 kd mannose 6-phosphate/IGF-II receptor from prelysosomes to the TGN in cell extracts. In vitro transport displays the same selectivity observed in living cells in that the transferrin receptor traverses to the TGN at a much lower rate than mannose 6-phosphate receptors. Furthermore, recycling of mannose 6-phosphate/IGF-II receptors to the TGN requires GTP hydrolysis and can be distinguished biochemically from the constitutive transport of proteins between Golgi cisternae by its resistance to the weak base, primaquine. PMID- 2971453 TI - Secretion of a suppressor cell inducing factor by an interleukin 3-dependent cell line with natural cytotoxic activity. III. Comparison with other interleukin 3 dependent cell lines. AB - We previously described the interleukin 3 (IL-3)-dependent cell line, M1-A5, which has both natural cytotoxic (NC) and suppressor cell activities, the latter of which is mediated, in part, by the release of two cytokines which activate suppressor cells from unprimed lymphoid precursor cells. In this study we have compared the M1-A5 cell line with four other IL-3-dependent cell lines to determine whether these dual activities are universally associated with IL-3 dependence and to test the hypothesis that there is a direct relationship between the cytotoxic and the suppressive activities. The cell lines tested were a bone marrow derived Dexter culture derived line (FDC-P1), two Moloney leukemia virus induced leukemias (DA-1 and DA-3), and a mast cell line (PT18(A17]. All lines were dependent on IL-3 for survival but FDC-P1, DA-1, and DA-3 showed varying degrees of short-term proliferation in granulocyte-macrophage colony stimulating factor (GM-CSF). The cell lines all expressed asialo GM1 and Ly-5 surface markers but differed with respect to other markers. DA-1 expressed MAC-1, FDC-P1 and DA-3 expressed Thy-1, and PT18(A17) expressed receptors for the Fc portion of IgE. The cell lines varied greatly in their cytotoxic activity against WEHI-164. FDC-P1, DA-1, and PT18(A17) had low NC activity. DA-3 had consistently high activity, greater than that seen with M1-A5 cells. However, none of the cell lines secreted constitutively a suppressor cell inducing factor (SIF). In addition, it was demonstrated that recombinant murine TNF did not activate suppressor cells capable of inhibiting antibody synthesis and that anti-TNF did not block SIF activity, thus suggesting that TNF contamination of the M1-A5 derived SIF preparation is not responsible for the induction of suppressor cells. We conclude that suppressor cell inducing factors are not universally secreted by IL-3 dependent cell lines, that there is no correlation between NC and SIF activity, and that the dual activities of M1-A5 cells are not mediated by TNF. PMID- 2971454 TI - Con A-stimulated thymocytes produce interleukin 2 after removal of Lyt-1 positive cells. AB - Mouse lymphocytes produce several lymphokines, including interleukin 2 (IL-2) and colony-stimulating factors (CSFs) following stimulation with T-cell mitogens. However, very little IL-2 is produced by thymocytes upon concanavalin A (Con A) stimulation. Strong selective inhibition of IL-2 production was observed when fresh spleen cells were mixed with Con A-activated thymocytes. Sorting of populations on the basis of antigenic phenotype showed that the cell mediating the blockage in IL-2 secretion is a large T cell expressing markers for both Lyt 1 and Lyt-2. This specific inhibition of IL-2 accumulation was not mediated by a soluble product, or by absorption on expressed IL-2 receptors on the activated thymocytes. Removal of the Lyt-1 positive cells from a thymocyte population renders it capable to produce IL-2 upon Con A stimulation, indicating a functional role of these cells. PMID- 2971455 TI - In situ localization of T cell receptor beta chain in the murine thymus: changes in the intrathymic distribution of thymocytes expressing beta chain during fetal development. AB - The expression of T cell receptor beta chain in the developing thymus was examined at the light and electron microscopic levels using the monoclonal antibody F23.1. Cells expressing cytoplasmic forms of beta chain were first observed at Day 16 of gestation, while thymocytes expressing cell surface beta chain were detected about a day later. Clustering of cortical F23.1+ cells was more pronounced in fetal thymus when compared to adult. The density of F23.1+ cells in the subcapsular areas of the thymus was initially lower than that in the rest of the cortex or the medulla. Within the subcapsular and cortical areas of the thymus there was an inverse relationship between the density of F23.1+ cells and cells labeled with the lectin from Dolichos bifloris, which binds to terminal alpha-linked N-acetylgalactosamine residues preferentially expressed by L3T4 /Lyt2- thymocytes. Although this pattern was less pronounced with increasing gestational age, it was still apparent at birth. PMID- 2971456 TI - T suppressor efferent circuit which affects contact sensitivity to picryl chloride: the late-acting, second nonspecific T suppressor factor bears I-A determinants which are responsible for the I-A genetic restriction in its interaction with its target cell. AB - The T suppressor efferent circuit in the picryl (TNP) system, which inhibits the passive transfer of contact sensitivity, involves at least two antigen nonspecific factors. The second nonspecific T suppressor factor (ns-2) bears I-A determinants of both the alpha and the beta chain as shown by affinity chromatography on immobilized anti-I-A monoclonal antibodies. Sequential absorption shows that the determinants of the alpha and beta chain occur on the same molecular complex. No absorption was obtained with anti-I-E antibody. There are two genetic restrictions associated with ns-2--the first is in its release from the second T suppressor efferent cell (on exposure to antigen) and the second is in its inhibitory interaction with its target cell. Both are MHC restricted and matching in I-A (but not I-E, or I-J) is sufficient. The question was asked whether the I-A of the ns-2 was directly responsible for the I-A genetic restriction in its action. F1 TsF was made in (H-2k X H-2b)F1 mice by injecting picrylated parental cells intravenously and triggering the release of ns-2 with the corresponding picrylated parental cells. Both I-Ak- and I-Ab positive ns-2 were produced and were separated by affinity chromatography on immobilized anti-I-A monoclonal antibody. The I-A phenotype of these separated ns 2 of F1 origin determines the genetic restriction in their action; i.e., I-Ak+ ns 2 only inhibits passive transfer by H-2k cells and I-Ab+ ns-2 only acts on H-2b cells. In contrast, the I-A haplotype of the picrylated cell used to induce the Ts cell which makes ns-2 is unimportant. It was concluded that the I-A on the ns 2, and not a possible recognition site for I-A, serves as a restriction element. This finding suggests that ns-2 may act directly on the I-A-restricted T cell which mediates contact sensitivity. PMID- 2971457 TI - Cell interactions in alveolar macrophage-mediated suppression of the immune response: an unusual suppressor pathway involving a population of T-cells that express Lyt-1, L3T4, and I-J. AB - Studies from this laboratory have demonstrated that incubation of murine alveolar macrophages (AM) with SRBC-primed spleen cells (SC) results in suppression of the in vitro plaque-forming cell (PFC) response and that suppression is mediated by a soluble factor contained in supernatants obtained from cultures of AM and SC. In the present study, immunological techniques employing monoclonal antibody (MoAb) were used to isolate various T-cell subsets in order to determine the phenotype of the cells which interact with AM to produce suppression. Spleen cell populations depleted of Thy-1+-, Lyt-1+-, L3T4+-, or I-J+-bearing cells failed to generate suppressive supernatants when cultured with AM. Depletion of Lyt-2+ T cells (the classical suppressor/effector subset) did not alter the ability of the remaining cell population to cooperate with AM for generation of suppressive supernatants. Direct suppression of the PFC response in cultures containing AM was abrogated after treatment of the spleen cells with anti-I-J, but not anti-Lyt 2 MoAbs. Reconstitution of the AM-mediated suppressive response with enriched populations of SC required the presence of T-cells which expressed Lyt-1, L3T4, and I-J. These results suggest the existence of an unusual suppressor pathway involving I-J restriction but which appears to be mediated by the interaction of AM with a population of T-cells that expresses surface markers characteristic of T-helper cells. PMID- 2971458 TI - The induction of antigen-specific thymic regulatory cells in the mouse. AB - Regulatory T cells are known to play a role in the termination of the immune response. In this investigation, we bring evidence in support of the induction of antigen-specific regulatory cells in the thymus of the mouse. These thymus derived regulatory cells depend upon injection with a high dose of antigen for their induction, appear in the thymus early after primary immunization (Days 7 12), are H-2- and strain-restricted in their action, and can by themselves suppress directly the secretion of antibodies from syngeneic antibody-forming cells in vitro. These regulatory cells, therefore, differ from previously reported murine suppressor T cells which exert their effect on antibody formation indirectly, generally by acting on helper T cells. PMID- 2971460 TI - Monoclonal antibody internalization and degradation during modulation of the CD3/T-cell receptor complex. AB - Although it is well known that the CD3/T-cell receptor (TCR) complex modulates from the surface of T cells upon exposure to monoclonal antibodies (mAb) directed against it, the fate of bound mAb has not been yet elucidated. We therefore perform direct binding experiments of 125I-labeled mAb against CD3 or TCR to investigate their fate in Jurkat T cells. We demonstrated that all mAb were progressively internalized and degraded in Jurkat T cells and that this degradation was inhibited by chloroquine, an inhibitor of lysosomal degradation enzymes. The sequestration of anti-CD3 mAb in acid compartments was furthermore shown using cytofluorometry. All together our results show that antibodies against CD3 or against TCR follow the same endocytic pathway. PMID- 2971459 TI - A soluble factor produced by bone marrow natural suppressor cells blocks interleukin 2 production and activity. AB - We previously reported that a population of Fc gamma-receptor+ (Fc gamma R+) suppressor cells present in normal unstimulated rabbit bone marrow inhibited the growth of autologous rapidly proliferating bone marrow cells devoid of Fc gamma R. It is now reported that the Fc gamma R+ bone marrow cells produced a soluble, nondialyzable suppressor factor(s) (SF) which blocked the proliferation of Fc gamma R- bone marrow cells. In addition, the Fc gamma R+ cells and SF significantly inhibited spleen cell proliferation in response to concanavalin A (Con A), phytohemagglutinin, and pokeweed mitogen. The bone marrow SF exhibited a dose-dependent suppression of the growth of IL-2-dependent T lymphocytes in the presence of IL-2. SF also completely blocked the production or release of IL-2 by Con A-stimulated T cells. Thus, these bone marrow natural suppressor cells produced a soluble factor, which regulated the growth of rapidly proliferating bone marrow cells and also regulated T cell reactivity by modulating IL-2 production and activity. PMID- 2971461 TI - [Contribution of ultrasonography and laparoscopy in the diagnosis of interstitial pregnancy]. PMID- 2971462 TI - [Medroxyprogesterone acetate in the treatment of recurrent vaginal candidiasis]. PMID- 2971463 TI - [Does therapy of hyperandrogenic conditions with cyproterone acetate improve the prognosis for growth in boys?]. PMID- 2971464 TI - The treatment of onychomycosis with a new form of tioconazole. AB - The difficulties encountered in the treatment of onychomycosis are primarily related to the necessity of prolonged systemic therapy. Many of these difficulties could, then, be avoided by the use of an effective local treatment. The present study compared the effectiveness and tolerability of two topical ungual preparations: a 28% solution of tioconazole and a 2% tincture of miconazole. The therapeutic results and tolerability of both preparations were found to be satisfactory. The tioconazole preparation proved to be slightly more effective although the difference was not statistically significant. PMID- 2971465 TI - Haemodynamic stability with midazolam-sufentanil analgesia in cardiac surgical patients. AB - Since the administration of both diazepam and midazolam are claimed to cause adverse haemodynamic effects following fentanyl or sufentanil intravenous injection, we evaluated the effectiveness and safety of the reverse sequence, (midazolam-sufentanil) on haemodynamic variables, adequacy of analgesia, amnesia and recovery in 15 adult patients undergoing coronary artery surgery (with a mean +/- SEM ejection fraction of 0.41 +/- 0.03). After routine premedication, midazolam 0.14 +/- 0.01 mg.kg-1 IV was given over one min followed 5 min later by sufentanil in incremental IV doses of 1.5 micrograms.kg-1 to a total pre intubation dose of 4.0-5.0 micrograms.kg-1 injected in 10 min. One minute after the initial dose of sufentanil, pancuronium 0.1 mg.kg-1 IV was given in 30 seconds. The incremental doses of sufentanil were based on a greater than 15 per cent increase in rate-pressure product. The mean dose of sufentanil before cardiopulmonary bypass was 9.6 +/- 2.1 micrograms.kg-1 and 13.9 +/- 1.3 micrograms.kg-1 for the entire procedure. A significant decrease in systolic and diastolic blood pressures occurred after midazolam administration which was sustained until sternotomy. A significant reduction in systemic vascular resistance occurred following midazolam. Sufentanil reduced the left ventricular stroke-work index. Tracheal intubation, skin incision and sternotomy elicited no adverse haemodynamic responses. Adequate analgesia, complete amnesia and early recovery of wakefulness were observed. PMID- 2971466 TI - Progress in the control of viral hepatitis: memorandum from a WHO meeting. AB - Viral hepatitis is a major public health problem in all parts of the world, with hepatitis B (HB) as the most important of all the viral hepatitides. It is estimated that worldwide there are nearly 300 million carriers of HB markers of active infection. More than 40% of persistently infected persons who survive into adult life will die of the consequences of HB, such as cirrhosis and hepatocellular carcinoma. The vaccines available against HB have an impressive record of safety and efficacy, and the best means of controlling the infection on a global scale, including the reduction of mortality due to its sequelae, will be by mass immunization of infants. In areas where most infections are acquired early in life the vaccine should be administered shortly after birth, and HB immunization should be integrated into the Expanded Programme on Immunization.Progress has been made also in developing hepatitis A vaccines and in isolating the agents that induce hepatitis non-A non-B. PMID- 2971468 TI - Parent playfulness: a case study of infant twins with handicaps. AB - This longitudinal case study was designed to provide descriptive data on game playing between one mother and father and their infant twin sons, each of whom had handicaps. Specific issues addressed were the proportion of interaction time spent playing parent/infant games, the types and characteristics of the games played, whether these varied between the mother and father, and whether there was a relationship between parental game playing and the developing but different abilities of the infants. Results showed many parallels with existing literature on parent/infant games and on differences between mothers and fathers. Results also indicated variations in game playing in relation to the extent of handicap in the infant. PMID- 2971467 TI - Potentiation of the antitumor activity of cisplatin in mice by 3-aminobenzamide and nicotinamide. AB - 3-Aminobenzamide (3AB) and nicotinamide (NA), inhibitors of adenosine-ribose transferase (ADPRT), potentiated the antitumor activity of cisplatin (DDP) on Ehrlich ascites carcinoma in mice. The mean survival times of the mice increased from 21.2-37.0 days in DDP-treated groups to 47.0-54.6 days in mice treated with DDP plus NA or 3AB. These drugs also potentiated DDP antitumor activity on sarcoma 180, with the inhibition rates increasing from 12.4%-20.8% in groups treated daily with DDP to 29.8%-46.4% in those treated with DDP plus NA or 3AB; however, neither 3AB nor NA alone showed any antitumor activity. The single-dose lethality of DDP on mice was partially reversed by either NA or 3AB. The pathological study revealed that the morphologic changes in the proximal tubules 1 month after a single dose of DDP (10 mg/kg) were partially prevented by a single protective dose (5 mmol/kg) of NA or 3AB. Our results suggest that the combination of DDP with ADPRT inhibitors might be used clinically in the future. PMID- 2971469 TI - The social skill difficulties of young adults with physical disabilities. AB - Adolescence is generally considered to be a time of increased social activity and the rapid expansion of a teenager's social circle. By contrast, a young person with a physical disability is likely to lack social independence, become socially isolated and experience difficulties in maintaining social relationships. The present study indicates that while both able-bodied and physically disabled young people experience difficulties in social situations, those with physical disabilities experience difficulties that are more severe in nature. Overall, 75% of the disabled group and 70% of the comparison group experienced difficulties in social situations. However, 37% of the disabled group experienced difficulties that were severe in nature compared to only 3% of the comparison group. Whilst these problems are of sufficient magnitude to warrant intervention, programmes of social skills training for people with physical disabilities are not yet generally available. It is recommended that training programmes in social skills should be made available to teenagers with physical disabilities. PMID- 2971471 TI - Excimer laser-induced simultaneous ablation and spectral identification of normal and atherosclerotic arterial tissue layers. AB - A krypton-fluorine excimer laser at a 248-nm wavelength was used to irradiate normal and severely atherosclerotic segments of human postmortem femoral arteries. Single pulses and multiple pulses required for penetration or perforation of the arterial wall were applied with 16 nsec pulse width and 5 J/cm2/pulse energy fluence. The total fluorescence of irradiated and ablated tissue was analyzed in real-time mode by means of spectroscopy. Each laser pulse produced one spectrum that was characteristic of the composition of the tissue layer, which was ablated. Fluorescence spectroscopy indicated a broad-continuum emission between 300 and 700 nm with peak fluorescence of equal intensity at wavelengths of 370 and 460 nm (ratio, 1.004 +/- 0.087) for normal media layers. Atheromas without calcification (lipid, fibrous, and mixed) were found with spectral maxima at the same wavelengths but with significantly reduced intensity at 460 nm (ratio, 1.765 +/- 0.263; p less than 0.001). In contrast to this broad continuum fluorescence, calcified plaques displayed multiple-line emission with the most prominent peaks at wavelengths of 397, 442, 450, 461, 528, and 558 nm. These fluorescence criteria identified the histologically classified target tissue precisely. Histological examination of the corresponding arterial layers indicated sharply delineated and circumscribed tissue ablation. These results indicate that simultaneous tissue identification (diagnosis) and ablation (treatment) by excimer laser irradiation is feasible under strict laboratory conditions. We conclude that this principle demonstrates the potential for laser beam control by means of target-specific ablation. PMID- 2971470 TI - Calcium uptake by sarcoplasmic reticulum isolated from hearts of septic rats. AB - Myocardial sarcoplasmic reticulum (SR) plays a critical role in the regulation of the cytosolic calcium fluctuations that occur during the cardiac cycle. One function of the SR is to lower the calcium concentration so that myocardial relaxation and thus ventricular filling can occur. The aim of the present study was to determine if hyperdynamic sepsis induced a decrease in the capacity of SR to take up calcium. This defect would result in decreased ventricular filling and thus decreased cardiac output, as has previously been shown in isolated perfused working hearts removed from septic rats. Therefore, rats were anesthetized with ether, and sepsis was induced by the injection of an aliquot of a fecal homogenate into the peritoneal cavity. Control animals either underwent surgery and received an aliquot of sterilized fecal inoculum (sham) or were untreated (no surgery). On day 2 after surgery, animals were anesthetized with pentobarbital, and hearts were removed, weighted, and SR isolated. The rate of uptake of 45Ca2+ by SR from septic rats was not depressed compared to controls but in fact was elevated. Maximum 45Ca2+ accumulated by the SR and Ca2+-stimulated ATPase activity were similar in SR from control and septic hearts. These results suggest that the contractile dysfunction noted in the myocardium in early sepsis is probably not due to inadequate SR removal of Ca2+ during diastole. PMID- 2971472 TI - Atrial natriuretic peptide distribution in fetal and failed adult human hearts. AB - The distributions of atrial natriuretic peptide (ANP) in human hearts during the developmental stage and in adult pathological states was examined with an antibody specific to human alpha-ANP. With immunoblotting and immunofluorescence methods, we found that a 17-kDa protein, which is a pro ANP, was expressed in human fetal ventricles, in which the numbers of myofibers containing ANP granules were more abundant in the subendocardial region than the subepicardial region. As determined by radioimmunoassay, the content of immunoreactive ANP (per milligram protein) in the developing heart was greatest in the left atrium and occurred decreasingly in the right atrium, right ventricle, and left ventricle, respectively. Because ANP content in the left ventricle declined during the progress of gestation in developing hearts and because it was very low, if ever detectable, in normal adult hearts, ventricular ANP expression appears to be developmentally regulated from the early gestational stage. However, it was reexpressed in the ventricles of patients who had suffered from severe congestive heart failure. In this situation, we found that the ventricular ANP expression was more marked in patients with dilated cardiomyopathy than in patients with severe valvular disease. Interestingly, in the ventricles of patients with dilated cardiomyopathy, ANP contents were higher in the left ventricular free wall than in the right ventricular free wall, although the left ventricular subendocardium contained more ANP than the subepicardium, showing a transmural gradient similar to that expressed in fetal ventricles. Thus, the expression of ANP in human ventricles is developmentally regulated from the early gestational stage, and even adult ventricular myofibers can synthesize ANP during severe congestive heart failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971474 TI - Dietary salt intake. A determinant of cardiac involvement in essential hypertension. AB - Because a given increase in afterload does not consistently produce the same degree of left ventricular hypertrophy, we evaluated several clinical, hemodynamic, and endocrine factors that are prone to modify the adaptation of left ventricular structure in patients with mild essential hypertension (World Health Organization stages I or II). Dietary salt intake assessed by sodium excretion over 24 hours was a powerful determinant of posterior wall thickness (r = 0.64, p less than 0.001), relative wall thickness (r = 0.67, p less than 0.001), and left ventricular mass (r = 0.37, p less than 0.05). In contrast, diastolic pressure, body mass index, hematocrit, and epinephrine were found to be weaker determinants of left ventricular structure (r = 0.31-0.40, p less than 0.05). A stepwise multiple regression analysis revealed that sodium excretion was the strongest predictor for posterior wall thickness (p less than 0.02) and relative wall thickness (p less than 0.05) independent of the other examined variables. These results identify dietary salt intake as a strong determinant of cardiac structural adaptation to a persistent increase in arterial pressure. Consequently, a high salt intake might aggravate and, conversely, dietary salt restriction might prevent (or at least mitigate) the development of left ventricular hypertrophy in patients with essential hypertension. PMID- 2971473 TI - Reversion of cardiac hypertrophy and reduced arterial compliance after converting enzyme inhibition in essential hypertension. AB - Blood pressure, forearm arterial hemodynamics (with a pulsed Doppler flowmeter), and echocardiographic parameters were studied in 16 patients with sustained essential hypertension before and 3 months after administration of the converting enzyme inhibitor perindopril. In a single-blind study versus placebo, it was shown that perindopril significantly reduced blood pressure (p less than 0.01), whereas there was an increase in brachial blood flow (p less than 0.01) because of a simultaneous increase in blood flow velocity (p less than 0.01) and arterial diameter (p less than 0.01). During a 5-minute period of wrist occlusion, blood flow velocity was reduced to a greater extent with perindopril than with placebo (p less than 0.001), whereas corresponding reductions in arterial diameter were equivalent, indicating that the increase in diameter after perindopril could not be explained simply on the basis of flow-dependent dilatation. During active treatment, brachial artery compliance increased (p less than 0.01) and pulse wave velocity decreased (p less than 0.01), whereas there was no change in the tangential tension of the arterial wall, defined as the product of mean arterial pressure and arterial diameter. Four weeks after treatment was stopped, blood pressure and forearm arterial hemodynamics returned toward baseline values. Cardiac mass was significantly decreased after perindopril (p less than 0.01) and remained decreased 4 weeks after cessation of treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971475 TI - Nonspecificity of a direct 17 alpha-hydroxyprogesterone radioimmunoassay kit when used with samples from neonates. AB - We analyzed 240 samples for 17 alpha-hydroxyprogesterone (17-OHP) with the direct assay kit ("Coat-A-Count" method for serum samples) from Diagnostic Products Corp. (DPC). The specimens were from 50 patients with known or suspected congenital adrenal hyperplasia (CAH); 74 mostly hospitalized neonates and infants, ages three days to three months; and 116 other patients, ages six months to 23 years. Samples from the CAH group were also analyzed with our in-house assay. For 39 of the neonatal samples, the analysis with the DPC assay was repeated with re-solubilized material that had been extracted from the serum with organic solvents. Values for "17-OHP" measured with the DPC direct assay were high, not only in CAH patients, but also in many of the unaffected neonates and infants. The extraction properties of the cross-reacting immunoreactive material into various organic solvent systems were different from those of 17-OHP, and were more like those of steroid sulfates. Because of this significant cross reactivity, we recommend that the DPC kit not be used for sera from children younger than six months of age, unless the method is modified to include an extraction step. PMID- 2971476 TI - Demonstration of monoclonal IgE by isoelectric focusing: first reported case of IgE myeloma in Australia. AB - A 56-year-old man who presented with back pain was diagnosed with multiple myeloma. Serum protein analysis by isoelectric focusing identified a monoclonal IgE lambda of unique spectrotype compared with other monoclonal immunoglobulins. The similarity of the spectrotype of this monoclonal IgE to that of two purified monoclonal IgEs suggests that these proteins have a characteristic spectrotype, sufficiently different from those of other immunoglobulin classes to allow its recognition. PMID- 2971477 TI - Estimation of nasal cross-sectional areas, using oral versus nasal pressure measurements. AB - This study examines calculations of model nasal cross-sectional area, using nasal versus oral pressure measurements. The results indicate that greater accuracy of nasal cross-sectional area estimation is achieved by using nasal rather than oral pressures. Nasal pressures measured in the anterior model nose more closely reflect nasopharyngeal pressures under a wide range of nasal constriction sizes and airflow rates. PMID- 2971478 TI - Enzyme immunoassay for alpha-human atrial natriuretic polypeptide--direct measurement of plasma level. AB - A sandwich enzyme immunoassay for alpha-human atrial natriuretic polypeptide (alpha-hANP) was developed. Polystyrene balls were coated with monoclonal IgG1 specific for the N-terminal half of the ring structure of alpha-hANP, and rabbit Fab' specific for the C-terminal (17-28) of alpha-hANP was conjugated to horseradish peroxidase. The polystyrene ball was incubated with alpha-hANP standards or plasma and subsequently with the conjugate. Bound peroxidase activity was measured by fluorimetry. No addition of hANP-free plasma for the standard curve was required when less than 50 microliters of plasma sample was used. The detection limit of alpha-hANP was 30 fg (10 amol). Using 50 microliters of plasma, we could detect 0.6 ng (0.2 pmol)/l without extraction. The basal plasma alpha-hANP level of healthy men in a supine position after an overnight fast was 24.5 +/- 13.2 (SD) ng/l and tended to decrease (15.3 +/- 8.5 (SD) ng/l) after intravenous administration of furosemide (40 mg) and subsequent one hour walking. There was a highly significant correlation between the plasma alpha-hANP concentrations measured by this enzyme immunoassay and by radioimmunoassay (r = 0.92). This enzyme immunoassay for alpha-hANP allows measurement of plasma alpha hANP without extraction even in the volume-contracted state. PMID- 2971479 TI - Measurement of platelet factor 4 and beta-thromboglobulin by an enzyme-linked immunosorbent assay. PMID- 2971480 TI - I-cell disease: evidence for a mannose 6-phosphate independent pathway for translocation of lysosomal enzymes in lymphoblastoid cells. PMID- 2971482 TI - Participation of suppressor-inducer cells in the suppression of adjuvant arthritis by transfer of spleen cells expanded by T cell growth factor. AB - We attempted to elucidate the mechanism of action of T cell growth factor (TCGF) expanded cells after stimulation with concanavalin A (Con A), a process which reduces the severity of adjuvant arthritis (AA), as seen in transferred syngeneic rats. TCGF-expanded cells were fractionated by Percoll density gradient or by the panning method, before the transfer. Transfer of cells with a density of between 1070 and 1079 suppressed AA most effectively, compared with other fractions with a density of less than 1070. A large number of the cells with density 1070-1079 were of the W3/25 phenotype. The transfer of W3/25 positive cells obtained by the panning method from TCGF-expanded cells reduced AA but not W3/25 negative cells. The suppressor function of TCGF-expanded cells was examined in an assay system in vitro. The addition of Con A-stimulated cells to the assay culture (but not expanded with TCGF), suppressed both the proliferation of spleen cells stimulated with Con A and the IgG production stimulated with lipopolysaccharide. The addition of TCGF-expanded cells to the assay culture had no effect on these responses. Thus, the W3/25 cells in the TCGF-expanded cells seem to function as suppressor-inducer cells, affect the presuppressor cells of the recipient rat and differentiate to suppressor effector cells. PMID- 2971481 TI - OKT4+ T cell deficiency and an association of immunoglobulin deficiency in systemic lupus erythematosus. AB - A patient with systemic lupus erythematosus and selective serum IgA and IgM deficiency also had a deficiency of OKT4+, OKT4A+, OKT4B+, and OKT4E+ T cells in circulating blood. This deficiency of helper/inducer T cell subsets in blood was associated with impaired T cell responses to phytohaemagglutinin in vitro, and an impaired ability to produce IgA and IgM by the patient's B cells. The patient's OKT4+ (CD4+) T cell deficient population suppressed IgM synthesis by normal B and T cells. PMID- 2971483 TI - A new experimental model of in-situ immune complex disease of the lung. AB - A model of acute immunological lung injury, initiated by the binding of passively administered antibody to an antigen (Concanavalin A (Con A] affixed to the pulmonary endothelium, is described. Pulmonary injury, monitored using the 125I albumin lung permeability index (LPI) did not occur with antigen deposition alone (LPI 0.235 +/- 0.012). The binding of antibody to the 'planted' lung antigen resulted in injury only when antibody binding exceeded a threshold value of 7.4 +/- 1.4 micrograms antibody globulin per gram of lung. Alevolar-capillary permeability changes were maximal 4 to 8 h after antigen-antibody interaction (LPI at 2 h, 0.353 +/- 0.015: at 4 h, 0.387 +0.33; at 8 h, 0.373 +/- 0.025; at 24 h, 0.289 +/- 0.031; at 48 h, 0.239 +/- 0.022). Lung injury was significantly attenuated by neutrophil (PMN) depletion (LPI at 4 h, 0.325 +/- 0.014: P less than 0.01cf PMN-intact animals), and further reduced by complement depletion (LPI at 4 h, 0.275 +/- 0.023: P less than 0.05 cf PMN-intact and PMN-deplete animals). This model of immune lung injury demonstrates the potential for in situ immune reactions on the pulmonary endothelial surface to induce acute inflammatory injury. Further the model illustrates the cooperative effects of neutrophils and complement in the genesis of inflammation, with both these mediator systems contributing to immunological pulmonary injury. PMID- 2971484 TI - Suppressor cell activity of human alveolar macrophages in interstitial lung diseases. AB - It has been shown that alveolar macrophages (AM) are able to modulate lymphocyte proliferation in vitro. The purpose of this study was to evaluate the effect of AM on the proliferation of autologous peripheral lymphocytes (APL) in patients with interstitial lung disease (ILD) compared to controls. Thirty patients were investigated: eight with idiopathic pulmonary fibrosis (IPF), nine with sarcoidosis(SA), seven with rheumatoid arthritis (RA) and six controls (CO). AM and APL were co-cultured at an increasing macrophage/lymphocyte ratio: 1%, 10%, 20% and 50%. A dose-dependent effect was observed and related to the number of AM added to APL, enhancing at low ratios and suppressing at high ratios. Suppression of proliferation by 50% AM differed in the four groups tested: 94.6% (92-98) in IPF, 73.0% (49-100) in SA, 43% (25-57) in RA, 32.4% (22-41) in the CO (P less than 0.01, P less than 0.05, and P0.05 respectively, compared to CO). Suppressive cell activity of AM from patients with ILD was higher than suppressive cell activity of the CO group. Suppression with 10% of AM in ILD group was 18% (2.2 62) compared with 2.58% (-13-17) in the CO group (P less than 0.05). 20% AM in ILD group showed 35% (3-76) suppression in comparison with 9.76% (-11-27) in the control group (P less than 0.01), 50% AM in ILD have a suppressive activity of 71% (25-100) in contrast to 32.4% (22-41) in control (P less than 0.01). In conclusion, AM from patients with interstitial lung diseases have a significantly stronger suppressive effect on the proliferation of autologous peripheral lymphocytes than controls. This is a new aspect of the study of activation of AM in these kinds of disorders. PMID- 2971485 TI - Rapid appearance of plasmin in tear fluid after ocular allergen exposure. AB - Specific allergens were applied topically to the conjunctivae of 18 allergic patients. In cases with positive allergic reaction, proteolytic activity, identified as plasmin using zymographic analysis and a monoclonal anticatalytic antibody to plasmin, was found to appear within 3-5 min in the tear fluid, reaching concentrations of 2.2-28.6 micrograms/ml. No plasmin was detected, either in the tear fluid of 30 unchallenged non-atopic control persons, or atopic patients provoked with a test material that had given no reaction in a skin prick test. These findings suggest that generation of plasmin is a consequence of the conjunctival allergic reaction. PMID- 2971486 TI - Correlation of immunoregulatory function with cell phenotype in cord blood lymphocytes. AB - The strong suppressor activity of cord T lymphocytes contrasts markedly with their mainly CD4 (helper) rather than CD8 (suppressor) phenotype. We studied the phenotype of cord CD3, CD4 and CD8 cells compared to adult cells using the monoclonal antibodies, 2H4, 4B4, and UCHL1. Almost all cord CD4 lymphocytes carried the suppressor-inducer marker 2H4, whereas 4B4+ UCHL1+ helper-inducer cells were virtually absent; CD8 cord cells were also of the 2H4+ 4B4- UCHL1- phenotype. In contrast in adult peripheral blood, half of the T cells, whether CD4 or CD8, were 2H4+ and half 4B4/UCHL1+. The suppressor-inducer phenotype of cord T cells was shown, in parallel functional experiments, to correlate with their enhanced proliferation to lectin and poor production of immunoglobulin and with the ability of cord mononuclear cells to suppress proliferation and immunoglobulin production by adult cells in co-culture experiments. These results indicate that the major imbalance in the cord CD4 subset in favour of 2H4 cells can explain many of the functional differences from adult cells. However, involvement of other cell types, in particular of the monocyte lineage, is necessary to explain other properties of immunocompetent cord cells. PMID- 2971487 TI - T cell clones providing helper function for IgE synthesis release soluble factor(s) that induce IgE production in human B cells: possible role for interleukin 4 (IL-4). AB - We have previously demonstrated that a proportion of human T cell clones (TCC) derived from tonsil or peripheral blood (PB) of non-allergic donors, upon triggering with phytohaemagglutinin (PHA) or anti-CD3 monoclonal antibody (MoAb), were able to provide help for IgE synthesis in B cells from both allergic and non allergic individuals. In this study we show that, upon PHA stimulation, culture supernatants from 10 selected TCC active on IgE synthesis also provided helper activity for IgE, whereas supernatants from unstimulated cultures of the same TCC were ineffective. In contrast, culture supernatants derived from five PHA stimulated TCC, unable to provide helper function for IgE synthesis, consistently failed to elicit production of IgE. While the induction of IgE synthesis by TCC occurred in B cells from virtually all allergic and non-allergic donors, their soluble factor(s) were found to be able to provide substantial help for IgE production only in B cells from a proportion of donors tested. In addition, B cells from non-atopic donors usually appeared to be less responsive than atopic B cells to the activity of such factor(s). In contrast, synthesis of both IgG and IgM was induced in every B cell donor by both TCC and their supernatants. Partial characterization of the factor(s) providing helper function for IgE synthesis in B cells showed that it apparently had a mol. wt between 10 and 50 kD and did not bind to immobilized IgE. Such an activity appeared to be associated with the presence of interleukin 4 (IL-4) in supernatants and it was inhibited by adding both gamma-interferon and anti-human IL-4 antibody in culture. PMID- 2971489 TI - Alterations in thymocyte subpopulations in Down's syndrome (trisomy 21). AB - To correlate the histologically observed thymic abnormalities with the cellular immunodeficiency found in Down's syndrome (DS), thymus fragments and thymocyte suspensions from 14 noninstitutionalized DS subjects were studied. Histologic examination and immunohistologic studies using an anticluster of differentiation (CD) 1 monoclonal antibody showed a contracted cortex due to cortical thymocyte depletion. When DS unselected thymocytes were phenotyped, a significant reduction of CD3-, CD1-, CD4-, and CD8-positive cells was found as compared to controls. To evaluate if the deficient expression of these markers was due to the reduction of thymocyte subsets identifiable on the basis of their physical properties, we separated DS unselected thymocytes into 10 fractions by continuous Percoll density gradient centrifugation. DS thymuses were almost completely devoid of high density thymocytes. Since in normal thymus, these cells correspond to small CD1+, CD4+, CD8+, and 50% CD3+ cortical thymocytes, their absence may explain the unrestricted reduction of markers on DS unfractionated thymocytes. Furthermore DS thymuses appeared to be enriched in CD1+ first fraction (Fr1) low density thymocytes of the Percoll gradient. Fr1 CD1+ cells constitute the main spontaneously proliferating pool in normal human thymus. When the spontaneous proliferating activity of DS Fr1 was compared to that of the control, a significant reduction was observed. This reduction associated with the absence of high density thymocytes, with the reduction of cells expressing alpha- and beta chains of the T cell receptor and in conclusion with the lymphocyte depletion, suggests that in DS thymuses there is a deficient expansion of immature T cells resulting in a reduction of the various thymocyte subpopulations, including the thymocyte pool able to differentiate into functionally mature T cells. PMID- 2971488 TI - T lymphocytes of the human colonic mucosa: functional and phenotypic analysis. AB - Normal human colonic lymphocyte populations were isolated for both phenotypic analysis by double-label immunofluorescence and assessment for regulatory effects on Ig production by co-culture with responder cells from colonic mucosa and peripheral blood. Mean CD4:CD8 ratios for colonic intraepithelial lymphocytes (IEL) and lamina propria lymphocytes (LPL) were comparable to values obtained from tissue sections. IEL alone did not produce Ig in vitro and were without effect on Ig production when co-cultured with LPL. However, T-enriched LPL had a marked helper effect for T-depleted LPL. Maximal help was for IgA production, increasing with numbers of T-enriched cells. In colonic LPL T-depleted and T enriched co-cultures, pokeweed mitogen (PWM) had no significant effect. By contrast, in co-cultures of T-enriched and T-depleted peripheral blood mononuclear cells, Ig production was PWM-dependent. In all experiments with colonic mucosal responder cells, IgG production was low. The effects of unfractionated colonic biopsy lymphocytes on T-depleted peripheral blood mononuclear cells were additive for IgM production and synergistic for IgA synthesis, although almost no IgG was produced. Moreover, PWM had helper effects for IgM, but was suppressive for IgA production. These data suggest that colonic mucosal regulatory cells reside in the lamina propria, and predominantly provide help for IgA and IgM synthesis. The data further suggest the existence of a pre stimulated IgA-specific T helper cell population. PMID- 2971491 TI - Effect of interferon-gamma on the abnormality of T cell activation in NZB mice. AB - It is known that New Zealand black (NZB) mice have a defect in the autoantigen recognition mechanism by T cells. The present study was carried out to examine whether the defect could be improved by normalizing the self Ia molecule expression on thymic reticuloepithelial cells (TRC) by using interferon-gamma (IFN-gamma). Syngeneic mixed lymphocyte reaction (SMLR) was used as an indicator of the mechanism of autoantigen recognition. In the reaction, T cells separated from the thymus were used as responders, and machrophages separated from the spleen were used as stimulators. When the responders were cultured with TRC, in which self Ia molecule expressions had been enhanced by IFN-gamma, for activation, the low level of SMLR in NZB mice was elevated close to the normal level. It was suggested, therefore, that IFN-gamma could normalize the defective autoantigen expression on TRC in NZB mice and improve the abnormal T cell activation against autoantigens. PMID- 2971490 TI - Rejection of bone marrow transplant and resistance of alloantigen reactive cells to in vivo deoxyadenosine in adenosine deaminase deficiency. AB - Severe combined immunodeficiency disease (SCID) in patients with adenosine deaminase (ADA) deficiency is thought to result from increased levels of purine metabolites. We attempted to immunosuppress a patient with ADA deficiency and SCID using a continuous infusion of deoxyadenosine to obtain engraftment of a T cell-depleted haplocompatible parental bone marrow graft. Before administering the drug in vivo, we investigated hematopoietic colony formation in two children with ADA deficiency (including the potential recipient), the obligate heterozygote donor (father), and normal controls using deoxyadenosine and erythro 9-(2-hydroxy-3-nanyl)adenosine (EHNA), and inhibitor of ADA. Deoxyadenosine alone in concentrations as high as 100 microM had no significant affect on erythroid (BFU-E) or myeloid (CFU-c) colony formation. However, in the presence of EHNA there was a significant reduction in BFU-E and CFU-c growth in all subjects and controls. Increasing doses of deoxyadenosine were given to one patient with ADA deficiency and SCID as a continuous 24-hr intravenous infusion. We found that there was a linear relationship between the dose administered and the plasma level; however, doses greater than 100 mg/day were required to increase erythrocyte dATP levels. We were able to raise intracellular dATP levels to more than three times baseline with doses of deoxyadenosine of 200 mg/day. However, there were no significant effects on the absolute lymphocyte counts or the lymphocyte responses to mitogen or alloantigen, and the haploidentical marrow failed to engraft. Our results suggest that the bone marrow of ADA-deficient patients is normal with respect to standard colony formation, that inhibitors of ADA do not adequately model the deficient state, and that the immunodeficiency in ADA deficiency is not proportionately related to either the deoxyadenosine or dATP levels, both of which were significantly elevated at the time of transplantation. PMID- 2971492 TI - Influence of mouse trisomy 16 on expression of specific genes. AB - We examined developmental changes in the relative activities of three different isozyme systems: aldolase, enolase and phosphoglycerate mutase, in tissues of fetal mice with trisomy 16 and of fetal euploid littermates. We wanted to determine whether morphological abnormalities such as reduced weight and size, which are generally observed in murine trisomy, are reflected at the molecular level. Following electrophoretic separation and subsequent measurement of relative activities of enolase isozymes in brain and phosphoglycerate mutase isozymes in heart, we found no significant differences between trisomy 16 fetuses and their euploid littermates. Synthesis of liver-specific aldolase was, however, delayed in trisomy 16 fetuses. PMID- 2971493 TI - Developmental consequences of autosomal aneuploidy in mammals. AB - Autosomal aneuploidy in mammals adversely affects developmental processes. In human beings, for example, trisomy 21 is the most frequent aneuploidy detected among newborns and the most common known genetic cause of mental retardation. In this review, several hypotheses are discussed that have been proposed to explain the mechanisms by which aneuploidy (especially trisomy) disrupts development. These mechanisms included specific gene dosage effects, generalized disruption of genetic homeostasis, and the influence of the parental origin of the duplicated chromosome. The availability of specific chromosomal rearrangements in mice, coupled with selective breeding schemes, permits generation of aneuploidy of specific chromosomes or chromosomal segments on controlled genetic backgrounds, thus enabling the systematic study of the causes and consequences of defined aneuploidy. Phenotypic characteristics associated with a number of specific aneuploidies in the mouse are discussed. Emphasis is placed on the effects of trisomy 16. Genetic homology between mouse chromosome 16 and human chromosome 21 has led investigators to suggest that analogous mechanisms will be responsible for the developmental abnormalities produced in these respective aneuploidies. Analysis of trisomy 16 mice from the organismal to the subcellular level has revealed a number of phenotypic characteristics (particularly neurobiologic ones) shared with human trisomy 21. The dosage effects of shared genes (or their products) may contribute to the development of these features. PMID- 2971494 TI - Sebaceous gland activity in black skin. AB - Sebaceous gland secretion was measured in 649 male and female subjects, of whom 67 (10.3 per cent) were black. No consistent difference in sebaceous gland activity was found between black and white skin. As sebum is an integral etiologic factor in acne, these findings are consistent with the clinical impression and with epidemiologic data, albeit scant, that the incidence of acne vulgaris in the black population differs little, if at all, from the incidence in the white population. PMID- 2971495 TI - Effects of acute smoking on the hemostatic system in humans. AB - Habitual smoking is one of the best established risk factors for cardiovascular disease. The pathogenesis of smoke-induced damage is not so well clarified, but it probably includes--among some other aspects--an activation of the hemostatic system. Recently it has been shown that smoking a single cigarette can significantly decrease the coronary blood flow in coronary patients as well as in normal subjects. We tested the hypothesis that the acute effects of smoke are mediated by the hemostatic system. Seven healthy male volunteers, aged 20-40 years (mean 32 +/- 6 years), entered the study. All were habitual smokers, but had abstained from smoking in the 12 hours preceding the test. After lying in absolute rest for about 30 minutes, each subject smoked a cigarette containing 1.2 mg of nicotine. Immediately before and after smoking, blood was drawn by clear venipuncture for the evaluation of the following hemostatic variables: collagen-induced platelet aggregation by the method of Born; thromboxane B2 (TxB2) production by platelets stimulated with collagen, radioimmunoassay (RIA); plasma beta thromboglobulin (TG) (RIA); plasma fibrinopeptide A (FPA) (RIA); plasma fibrinolytic activity in the euglobulin fraction (NEF) (fibrin plate method). The following results, respectively before and after smoking, were observed: collagen-induced platelet aggregation 55 +/- 3 vs. 57 +/- 6%; TxB2 100.5 +/- 5.9 vs. 90.3 +/- 9.0 ng/10(8) platelets; plasma beta-TG 20.8 +/- 2.2 vs. 19.2 +/- 2.3 ng/ml; plasma FPA 2.3 +/- 0.3 vs. 2.2 +/- 0.1 ng/ml; NEF, lysis diameter 16.8 +/- 1.6 vs. 16.7 +/- 1.7 mm; NEF + C1 inhibitor lysis diameter 8.8 +/- 0.7 vs. 9.1 +/- 0.7 mm.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971496 TI - Inhibition of polymorphonuclear leukocyte chemotaxis by the mucoid exopolysaccharide of Pseudomonas aeruginosa. AB - We tested the effect on human polymorphonuclear leukocyte (PMN) chemotaxis of the mucoid exopolysaccharide (MEP) elaborated by Pseudomonas aeruginosa strain P1M recovered from the sputum of a patient with cystic fibrosis (CF). A dose-related inhibition of chemotaxis under agarose was observed when mucoid exopolysaccharide at concentrations of 0.5 to 5.0 mg/ml was incorporated into the agarose. Similar inhibition was observed with the closely related polysaccharide alginic acid, but not with the neutral polymer Ficoll. The exopolysaccharide did not bind or inactivate the chemoattractant substance, nor was it toxic to PMN's. Digestion with alginase did significantly abrogate the inhibitory effect of alginic acid, but not of MEP. There was no difference in chemotaxis in the presence of the exopolysaccharide preincubated with heat-inactivated pooled antisera from CF patients chronically colonized with mucoid P. aeruginosa compared to MEP alone, or MEP preincubated with heat-inactivated pooled normal human sera. Inhibition of chemotaxis may represent yet another pathogenetic property of P. aeruginosa mucoid exopolysaccharide. PMID- 2971497 TI - Decreased iodine-123 IMP caudate nucleus uptake in patients with Huntington's disease. AB - To determine whether I-123 isopropyl iodoamphetamine (IMP) uptake is reduced in the basal ganglia of patients with Huntington's disease compared with that in aged-matched normal and abnormal control subjects, a caudate ratio was defined that compared the average separation (in pixel units) between the midline and the left and right caudate heads to the width of the brain as measured on transaxial cross-sections of I-123 IMP SPECT brain images. For six patients with Huntington's disease, the average caudate ratio was 29.0% (SD +/- 2.7%), significantly higher than that for 12 normal volunteer subjects (average caudate ratio, 19.1% +/- 3.5%; p less than 0.001) and 13 patients with a variety of other neurologic disorders (average caudate ratio, 19.3 +/- 2.2%; p less than 0.001). PMID- 2971498 TI - Tolerance of once-daily dosing of netilmicin and teicoplanin, alone or in combination, in healthy volunteers. AB - The purposes of this study were to test the pharmacokinetics and renal and otologic tolerances of a once-daily regimen of netilmicin and teicoplanin administered intramuscularly, alone or in combination (4.5 and 6 mg/kg, respectively), for 7 days in 30 healthy male volunteers. Teicoplanin induced only a mild increase in enzymuria. Nephrotoxicity was moderate and reversible with netilmicin; there was increased enzymuria and alteration in diluting ability, without significant changes in urinary beta 2-microglobulin levels, concentrating ability, and glomerular filtration rate. Ototoxicity was not detected in any of the subjects. Our results suggest that (1) teicoplanin and netilmicin given once daily induced only slight, reversible tubular damage, without any sign of ototoxicity; (2) their combination was not more toxic; and (3) clinical studies can be envisaged to evaluate the efficacy and tolerance of once-daily regimens in long-term treatment. PMID- 2971499 TI - Presumptive evidence for an immunosuppressor susceptibility gene, linked to HLA, in rheumatic fever. AB - In a study of rheumatic carditis, 135 members of 21 multiplex families have been investigated, along with 60 normal unrelated control individuals. Circulating T lymphocytes were reduced (as a percentage of total blood mononuclear cells) in all 'rheumatic' individuals, in 7 of 40 normal parents, and in 8 of 49 normal sibs. An immune response characterized by an increase in the proportion of suppressor T cells occurs in most individuals affected by rheumatic carditis and this change persists for a long time. Genetic analysis revealed three important points: 1. increased HLA haplotype sharing amongst the affected sib-pairs; 2. the possibility of using low circulating T-cell percentage as a marker of susceptibility; 3. presumptive evidence for a recessive susceptibility gene linked to HLA and responsible for the suppressor cell response. PMID- 2971500 TI - Histamine-like reaction after a single intravenous dose of rifampin. PMID- 2971501 TI - Antimicrobial agent resistance in Neisseria gonorrhoeae in St. Paul Minnesota. AB - In a prospective survey of 283 Neisseria gonorrhoeae isolated in St. Paul, Minnesota, 5% were found to produce beta-lactamase, and 11% were resistant to penicillin by chromosomal mediation, thus indicating a hyperendemic level of resistance according to Centers for Disease Control guidelines. A significant level of chromosomal resistance (30%) was noted for tetracycline, and MICs for spectinomycin approached the upper limits of susceptibility for a large number of isolates. The need for uniformity in agar dilution test methodology is discussed, and the variability in antimicrobial agent interpretive breakpoint criteria is illustrated by comparing penicillin breakpoints published by the Centers for Disease Control with those of the National Committee for Clinical Laboratory Standards as applied to the study isolates. PMID- 2971502 TI - A program for the identification of steady-state processes in biological systems. AB - A dedicated nonlinear regression program for the identification of steady-state processes is described in detail. Experimental data are fitted to the rational function which describes such systems. The algorithm makes use of parameter separation to relieve the user from the need to assign initial estimates to the linear parameters and to speed up the computation. A modified Marquardt algorithm is used and some properties of the function are exploited to improve the convergence rate and execution time. The search is made constrained by the addition of a penalty function to the residual sum of squares. Graphics are included to help the user find good initial estimates for the nonlinear parameters and as an aid to judge the goodness of fit of the result. Residual plots are also available to help the user identify the origin of the differences in the statistical tests. A final prediction error test is provided since this statistic is thought to be the most appropriate for the nonlinear function used. An enzyme kinetic example illustrates the procedure. The program was coded in Turbo Pascal on an IBM compatible PC. PMID- 2971503 TI - Reduced anaerobic glycolysis in oral contraceptive users. AB - Oral contraceptives containing combinations of estrogens and progestogens are known to impair glucose tolerance. The biochemical mechanisms underlying this lesion are speculative. In the present study women treated with OC for periods exceeding 10 cycles showed significant reduction in the activity of the key glycolytic enzyme phosphofructokinase (40%) and the levels of lactate (42%) in the erythrocytes compared to controls. These observations in women are analogous to those made earlier in female rats. PMID- 2971504 TI - Assessing changes in vaginal bleeding patterns in contracepting women. AB - In clinical trials comparing alternative contraceptive methods, women often discontinue for disturbances in menstrual bleeding patterns. Vaginal bleeding diaries have been used to monitor these patterns, and this paper suggests two new methods of analysis of these data. One analysis measures trends over time in the occurrence of specific patterns and, a second analysis, the relation between a woman's current experience and the probability of continuing method use. Contrasts between these approaches and the reference period method of analysis are made. A randomised clinical trial of the relative efficacy of 100 and 150 mg doses of depot-medroxyprogesterone acetate (DMPA) is used for illustration. PMID- 2971505 TI - Vaginal bleeding patterns among women using one natural and eight hormonal methods of contraception. AB - Menstrual diary records were obtained from a total of 5257 women using nine different methods of contraception, one natural and eight hormonal. This paper presents a comparative analysis of their vaginal bleeding patterns. The analytic procedures follow the recommendations of a recent WHO workshop on bleeding pattern analysis, which involve dividing each subject's diary into successive 90 day reference periods, calculating ten indices for each period, and classifying women according to whether they have "clinically important" bleeding disturbances. In general, the findings of this analysis confirm those of previous studies. Women using the natural method, who were deliberately selected for the regularity of their menstrual cycles, averaged three bleeding/spotting episodes of length 5 days in each 90-day period, with very little variability within or between women. Subjects given a combined oral contraceptive had more regular patterns than any other treated group, with short (4-day) episodes and 23-24 day bleeding-free intervals. Progestogen-only pill users had more frequent, longer episodes and shorter, less predictable intervals than combined pill users. Contrary to widely-held beliefs, the progestogen-only pills produced fewer spotting days than the combined pills, and almost no spotting episodes at all. Nearly half of vaginal ring users experienced some menstrual disturbance in each period; their most common problems were irregular, infrequent or prolonged bleeding. Women using the long-acting injectable, depot medroxyprogesterone acetate, had totally unpredictable patterns, with infrequent but prolonged bleeding/spotting episodes. The incidence of amenorrhea rose from just under 10% in their first injection interval to over 40% in their fourth. The methods of analysis recommended by WHO in 1985 still require substantial refinement. Nevertheless, they are more sensitive than those used previously for WHO trials and produce an easily understood, clinically meaningful characterization of bleeding patterns. PMID- 2971506 TI - The association between vaginal bleeding patterns and reasons for discontinuation of contraceptive use. AB - This paper describes the relationship between menstrual bleeding patterns recorded and reasons for discontinuing method use given by women using one of four types of hormonal contraception: combined oral pills, progestogen-only oral pills, a vaginal ring or depot-medroxyprogesterone acetate (DMPA). The women were recruited to five clinical trials, each of which lasted at least 48 weeks. The subject's reason for discontinuation was recorded if she stopped contraceptive use before the scheduled time. Overall, the reason expressed for discontinuing method use was a close reflection of the subject's experience. Among women using either type of oral contraceptive or a vaginal ring, subjects who discontinued for a non-menstrual reason or were lost to follow-up had bleeding patterns which did not differ markedly from those of women who continued method use. Only in the DMPA group was there any evidence that women who complained of non-menstrual side effects or were lost to follow-up might have ceased method use because they were unwilling or unable to tolerate their bleeding patterns. Subjects who discontinued because of amenorrhea had few bleeding/spotting days and at least one prolonged bleeding-free interval; women who complained of longer bleeding had long episodes and short intervals. Women who reported heavier bleeding had patterns which were similar to, but less extreme, than those of subjects who reported longer bleeding. The complaint of irregular bleeding, however, did not concur with the bleeding patterns recorded, regardless of contraceptive method. The predominant menstrual disturbance in this group of women was frequency of bleeding. They had no special difficulty in predicting either the time of onset or the length of their bleeding episodes. There were marked differences between individuals in terms of their acceptance of bleeding disturbances. Nevertheless, the results of this study confirm the importance of counselling. Women using DMPA tolerated far greater menstrual disruption than subjects using any other method. Subjects using an oral contraceptive were unlikely to have been warned of potential bleeding problems; when they encountered any, they tended to cease method use. In contrast, subjects using DMPA would have been advised to expect irregular patterns and possibly amenorrhea; prepared for such disturbances, their perseverance was remarkable. PMID- 2971507 TI - Determinants of menstrual bleeding patterns among women using natural and hormonal methods of contraception. I. Regional variations. AB - Records of the occurrence of vaginal bleeding were obtained from women using either a natural method of contraception or one of four types of hormonal contraceptive. The relationships between their bleeding patterns and a number of demographic variables were examined, with the aim of identifying subgroups of women who, if they used a particular hormonal method of contraception, would be likely to suffer more or less disruption to their bleeding pattern than the 'norm'. Within contraceptive method, bleeding patterns were more closely related to the women's geographical region of residence than to any other factor. Some of the differences between regions were consistent across contraceptive methods. European women tended to have more bleeding/spotting days than women in other regions; Latin American women had relatively short episodes and long bleeding free intervals, whether they were using the ovulation method, combined pills or a vaginal ring. Other differences were method-specific. Women using combined pills in India or Pakistan had fewer spotting episodes than women using the same method elsewhere; those using progestogen-only pills had more. Regional variations in bleeding patterns were particularly marked among women using DMPA, and increased over time: by their fourth injection interval, 25% of European women had amenorrhea, as compared with 72% of subjects in North Africa. These findings need to be confirmed by carefully controlled studies of menstrual bleeding patterns and their acceptability in various ethnic groups. The results would be valuable in counselling new contraceptive acceptors, and could eventually guide the choice of methods for introduction into national family planning programmes. PMID- 2971508 TI - Determinants of menstrual bleeding patterns among women using natural and hormonal methods of contraception. II. The influence of individual characteristics. AB - Menstrual diary records were obtained from women using either a natural method of contraception, a combined or progestogen-only oral contraceptive, a vaginal ring, or a long-acting injectable (DMPA). Within each method group, multiple regression analysis was used to examine the relationships between the subjects' bleeding patterns and their age, age at menarche, ponderal index, obstetric and contraceptive history and ethnic origin. The influence of ethnic origin was described in the preceding paper. Among combined pill users, increasing age was associated with more frequent spotting episodes. In both the untreated and combined pill groups, women with a higher ponderal index had less variable bleeding-free intervals; in the vaginal ring and DMPA groups, the more obese women had shorter bleeding/spotting episodes and longer bleeding-free intervals. Among combined oral contraceptive users, the most influential variable was the time since the end of the woman's last pregnancy: subjects who had been more recently pregnant had longer, less predictable episodes and shorter intervals. Previous oral contraceptive use was associated with more predictable bleeding patterns among women currently using either type of oral pill. In the vaginal ring and DMPA groups, subjects whose last pregnancy had ended in abortion had more bleeding/spotting days and episodes than those who had had a live birth. As reported previously, bleeding patterns were more closely related to the woman's geographical region of residence than to any other factor. The associations found with other individual characteristics were often inconsistent or difficult to interpret. A number of variables which could potentially influence menstrual bleeding patterns, such as nutritional status, were not measured and therefore could not be included in the analysis. However, contraceptive method and ethnic origin may be predominant influences, overriding any other factor. PMID- 2971509 TI - Serum levels of 3-keto-desogestrel and SHBG during 12 cycles of treatment with 30 micrograms ethinylestradiol and 150 micrograms desogestrel. AB - The serum concentrations of 3-keto-desogestrel (KDG) have been determined radioimmunologically in 11 female volunteers on Day 1, 10, and 21 of the 1st, 3rd, 6th, and 12th cycle of treatment with 30 micrograms ethinylestradiol and 150 micrograms desogestrel during the first 4 hours and 24 hours after intake. On the first day of each cycle the KDG levels were low, but increased thereafter until Day 21. Highest serum concentrations were measured on Day 21 of the 3rd and 6th cycle with peak levels between 1.5 and 6.2 ng/ml. Contrary to this, the KDG levels were significantly reduced during the 12th treatment cycle. The serum concentrations of SHBG rose significantly between Day 1 and Day 21 of each cycle reaching values which were 3-fold of those at the beginning of treatment. During the pill-free intervals, SHBG levels decreased but remained elevated as compared to controls. There was a significant correlation between the SHBG levels and the area under the KDG-concentration-versus-time curves (AUC) indicating a pronounced influence of the serum steroid-binding protein upon the pharmacokinetics of KDG. There were great interindividual differences in the KDG levels. The serum levels of the individual woman remain, however, in a relatively constant range throughout the treatment period of 12 months, possibly due to genetic factors. PMID- 2971510 TI - Complement in the pathophysiology and diagnosis of human diseases. AB - Complement is a humoral effector system composed of 21 plasma proteins that was identified initially because of its cytolytic effects. In addition to cytolysis, complement has a number of different functions related to inflammatory and other host defense processes. The description of the reaction mechanism includes: (1) activation of the classical pathway through recognition of IgG and IgM antibodies by C1q, (2) activation of the alternative pathway which is usually achieved without participation of immunoglobulins, (3) generation of proteolytic enzymes composed of heteropolymers that cleave certain precursor proteins, (4) formation of the membrane attack complex (MAC), and (5) participation of control mechanisms. Methodologies for studying protein concentration and functional activities of complement components include not only the classical hemolytic techniques but also the extremely sensitive new radioimmunoassays and enzyme immunoassays for measuring the products of complement activation that are generated in vivo. Examples of genetically controlled complement deficiencies have been published for most complement components. The symptomatology of some of these patients serves to emphasize the protective role of complement. Acquired deficiencies are significant not only as laboratory aids in diagnosis and to evaluate the course of certain diseases, but also to indicate possible pathogenic disease mechanisms. Recently, it has been recognized that the complement proteins with genes located in the HLA region are polymorphic. Certain variants of proteins C2, C4, and factor B occur with higher frequencies in certain diseases than in the general population, which appears to be of great practical importance in laboratory medicine. PMID- 2971511 TI - Highlights of the AAD post-graduate course "Recent Developments in Contact Dermatitis and Occupational Dermatology" sponsored by the AAD with the North American Contact Dermatitis Group San Diego, May 21-28, 1988. Part II. PMID- 2971512 TI - A comparison of Cleocin T Solution, Cleocin T Gel, and placebo in the treatment of acne vulgaris. AB - A twelve-week study in which investigators were blinded to the agents studied included 135 patients (aged twelve to thirty-five years) with moderately severe acne vulgaris. Patients were randomized into groups to compare the efficacy and local tolerance of 1 percent clindamycin phosphate gel, 1 percent clindamycin phosphate solution, and the gel vehicle which served as a placebo. One hundred and twenty-eight patients who completed at least six weeks of treatment were evaluated for efficacy of these agents. Both the solution and the gel were equivalent in reducing papule and pustule counts and results were superior to those produced by the placebo. All three treatments were tolerated well. PMID- 2971513 TI - Concanavalin A induced suppressor activity exerted by peripheral blood mononuclear cells--with special reference to chronic inflammatory bowel disease. PMID- 2971514 TI - The duration of action of the combination of fenoterol hydrobromide and ipratropium bromide in protecting against asthma provoked by hyperpnea. AB - We compared the duration of the protective effect of two beta-adrenoceptor agonists, fenoterol (200 micrograms) and salbutamol (200 micrograms), the anticholinergic agent ipratropium (80 micrograms), and the combination of fenoterol (200 micrograms) and ipratropium (80 micrograms) against challenge by eucapnic voluntary hyperventilation (EVH). Twelve patients with asthma performed EVH for two or four min at 60 percent maximal voluntary ventilation, 30 min, 2 and 4 h after treatment. All treatments (Rx) produced significant bronchodilation after 30 min. The Rx containing a beta-adrenoceptor agonist maintained this bronchodilation for at least 2 h. While all the Rx with a beta-adrenoceptor agonist significantly reduced the fall in forced expiratory volume in one second after EVH at 30 min, only the combination of fenoterol and ipratropium provided significant protection after 2 h. We advise that the duration of protective effect of beta-adrenoceptor agonists is short and patients with moderate to severe exercise-induced asthma may be better controlled by combination therapy. PMID- 2971515 TI - Should ipratropium bromide be added to beta-agonists in treatment of acute severe asthma? AB - In a double-blind randomized trial, 40 patients with acute severe asthma were given either nebulized salbutamol, 5 mg, or salbutamol, 5 mg mixed with ipratropium bromide 500 micrograms, on admission to hospital and again two hours later. There was no significant difference between the mean peak flows of the two treatment groups at any time. However, two hours after each treatment, there were fewer subjects in the ipratropium and salbutamol group whose peak flow rates had fallen back toward baseline levels than in the salbutamol only treatment group. Thus, although ipratropium did not improve the overall maximal bronchodilator response, it may have prolonged the duration of the response, which would be a clinically useful effect. PMID- 2971516 TI - Concord Chronicle. A special newspaper for special children. PMID- 2971517 TI - [The role of NMR in the diagnosis of lumbar disc pathology]. PMID- 2971519 TI - [The incidence of occupationally-induced allergic skin diseases in a large flower market]. AB - 150 questionnaires as well as epicutaneous tests in 56 individuals from a total of 675 persons cultivating and selling ornamental plants at the largest German flower market revealed that half of those investigated were suffering from allergic contact dermatitis. The leading plant species with sensitizing properties was found to be the chrysanthemum, followed by tulips and Alstroemeria cultivars. Allergic reactions to daffodils and primulas were rarely observed. Most of the reactions obtained with other Compositae species such as arnica, marguerite, sunflower, tansy and yarrow must be interpreted as cross-reactions due to the fact that cross-reactivity predominates within the sesquiterpene lactone constituents of the various Compositae species. PMID- 2971518 TI - Immune responsiveness to retinal S-antigen and opsin in serpiginous choroiditis and other retinal diseases. AB - The immune responsiveness to bovine retinal S-antigen and opsin has been investigated in some retinal disorders by means of in vitro lymphocyte proliferation, leukocyte migration inhibition and enzyme linked immune sorbent assays (ELISA). Sensitisation to S-antigen was observed in serpiginous choroiditis, but not in acute posterior multifocal placoid pigment epitheliopathy (APMPPE) or retinitis pigmentosa. No significant immune responsiveness was detected to opsin in any of the three diseases. Elevated antibody titers to S antigen were observed in some individual patients and healthy subjects. However, none of the patient groups exhibited an elevated antibody titer as compared to the control group. Although serpiginous choroiditis and APMPPE share some prominent clinical characteristics, the sensitisation in the former disease may perhaps be attributed to more severe and prolonged damage of the photoreceptor cells and blood-retina barrier. A combination of previous and present results suggests that in immunological investigations of retinitis pigmentosa patients it is more effective to use human than bovine S-antigen as test antigen because a species specific epitope seems to be involved. PMID- 2971520 TI - [Occupationally-induced allergic and photoallergic reactions to epoxide resins in railroad employees]. AB - Medical examinations were performed on 74 inductive oil wiring workers in contact with synthetic resins. 22 persons from a control group were examined. In the history, frequent complaints about the skin and mucous membranes were obtained, and clinically, moderate skin changes were observed in 9 individuals. These manifestations took the form of erythema and papules, appearing mainly on the uncovered parts of the body during the hot resin's hardening. In some of the workers, positive epicutaneous patch tests with cadmium, cobalt and epidian-3 and -5 were observed. Additional positive reactions were obtained via photo-patch tests with resins and solvents, indicating their photo-allergenic properties in some conditions. PMID- 2971521 TI - Adverse reactions of externally applied drugs and inert substances. AB - The lack of a first-pass effect for locally-applied substances is often not considered. This is especially true for such central nervous system agents as salicylic acid, hexachlorophene or caffeine which have a relatively low toxicity when applied orally. Manifestation of dermal irritation, such as a beginning rash or inflammation, are often tolerated because the beneficial effects of some topically applied substances outweight the damage (Dithranol, 5-Fluorouracil). This applies to some anti-phlogistics as well, which may also cause skin irritation. Some substances, such as benzoyl peroxide, vitamin-A acid and beta estradiol have direct or indirect desiccative effects so that skin irritations may occur. Other substances, such as tar, for example, are applied locally although they may have a carcinogenic potential when misused, or when contraindications for its use are disregarded. Local risks can be recognized in clinical-experimental and clinical testing of dermatological products. One must, however, look carefully for an intensification of possible risks related to the method of application, the condition of the skin, the site of application, supporting therapeutic measures or changes in the composition of the vehicle. Lanolin, cetyl alcohol and myristyl alcohol, sorbitol, isopropyl-myristate as well as polyethylene glycols (PEG) penetrate the skin like active substances. Some have been connected with skin allergies. In addition, some preservatives have mutagenic properties. Many vehicles cause dehydration of the horny layer and thus result in chronic surface damage. This is true for hygroscopic substances such as PEG as well as for liquid paraffins, lipid solvents (alcohol, acetone), solvents such as propylene glycol and for some O/W emulsifiers. PMID- 2971522 TI - [Functional relations between liver and kidney. I. Sodium excretion]. PMID- 2971523 TI - [Natriuretic hormones of the heart]. PMID- 2971524 TI - In vitro metabolism of 7 alpha-methyl-19-nortestosterone by rat liver, prostate, and epididymis. AB - The in vitro metabolism of 7 alpha-methyl-19-nortestosterone (7 alpha-methyl-3 oxo-estr-4-ene-17 beta-ol) was investigated in male rat liver, ventral prostate, and epididymis. Three metabolites were recovered from the liver incubation: 7 alpha-methyl-estr-4-ene-3,17-dione, 7 alpha-methyl-5 beta-estrane-3,17 beta-diol, and 7 alpha-methyl-3-oxo-estr-4-ene-16,17 beta-diol, whereas prostate and epididymis did not perceptibly metabolize the steroid. The kinetics of metabolism in liver differed significantly from those of testosterone or 19-nortestosterone. About 85% of the 7 alpha-methyl-19-nortestosterone added was recovered at the end of 30 min of incubation. We conclude that the androgenic potency of 7-alpha methyl-19-nortestosterone is not dependent on its further conversion to 5 alpha dihydro-metabolites. PMID- 2971525 TI - The effect of inhibition of aromatase enzyme activity on Leydig cell number and ultrastructure in beagles. AB - We have shown previously that administration of an orally active competitive aromatase inhibitor 4-(5,6,7,8-tetrahydroimidazo [1,5a] pyridin-5-yl) benzonitrile monohydrochloride to adult male beagles increases peripheral blood LH and testosterone concentrations, and that testes from treated dogs produce more testosterone when perfused in vitro than age-matched controls. In the present study we posed the question of whether the increased testosterone secretion by testes from these same aromatase-treated dogs was due to Leydig cell hypertrophy or hyperplasia, and if the latter, whether cytoplasmic organelles are increased. Beagles were treated with the inhibitor at a dosage of 2.5 mg/kg.day for 25 weeks and were euthanized by an overdose of iv sodium pentobarbital; testes were perfusion-fixed, embedded, and sectioned for stereological analysis. There were no significant differences in testis volume and absolute volumes of seminiferous tubules, blood vessels, lymphatic space, macrophage cells, and mesenchymal cells between the control and treated dogs. In contrast absolute interstitium volume and the absolute volume of Leydig cells per testis were significantly increased (P less than 0.05) in treated dogs. This increased Leydig cell volume per testis was due to increased volume of individual Leydig cells rather than to increases in Leydig cell number per testis. Additional studies showed that the surface area per Leydig cell of smooth endoplasmic reticulum, outer and inner mitochondrial membranes, and membranes of lipid droplets per testis were significantly higher (P less than 0.05) in the treated dogs as compared to the controls. In summary, the results of this study lead us to conclude that aromatase inhibition in the mature dog causes Leydig cell hypertrophy rather than hyperplasia and increased surface area per Leydig cell of subcellular organelles that contain enzymes involved in steroid biosynthesis. PMID- 2971526 TI - Progestin-mediated changes in progesterone receptor forms in the normal human endometrium. AB - Monoclonal antibodies to the human progesterone receptor (PR) were used to detect changes in PR form during the menstrual cycle. In proliferative phase samples, two PR proteins with mol wt (Mr) of 116,000 and 81,000 were detected on protein blots probed with anti-PR monoclonal antibodies. The 116,000 Mr protein was comprised of triplet isoforms, while the 81,000 Mr protein was a singlet. By contrast, protein blot analysis of secretory phase samples revealed a doublet isoform at 116,000 Mr and an 81,000 Mr singlet. Organ culture of human endometrium was used to mimic these changes in PR form in vitro. Although the triplet to doublet conversion was not realized in organ culture, time- and dose related changes in PR form were achieved which were similar to the in vivo state. Furthermore, these changes preceded the progestin-mediated induction of the enzyme estradiol dehydrogenase in parallel cultures, demonstrating that this is a useful system in which to study the relationship between receptor modulation and initiation and maintenance of biological response. PMID- 2971527 TI - The importance of variant insertions of the ligamentum teres hepatis in the Cruveilhier-Baumgarten syndrome. AB - Within the framework of a prospective study, we were able to establish that in 50 per cent of 220 patients submitted to laparoscopy, the ligamentum teres hepatis did not run from the porta hepatis directly to the umbilicus, but to a point of insertion craniad to the umbilicus in the median line of the anterior abdominal wall. As a rule, this topographic variant has no clinical relevance. It is, however, of importance in patients with cirrhosis of the liver and portocaval collateral channels. In our group, 25 per cent of the patients revealed complete cirrhotic transformation of the liver; in a quarter of these, the Cruveilhier Baumgarten syndrome presented, and thus also the possibility that large-caliber porto-femoral "umbilical" vessels might be running a course to the left of the umbilicus. The danger of injuring such a vessel during laparoscopy can, however, be prevented: in seven out of eight patients with portofemoral collaterals, this situation was established by ultrasonic examination, and in three cases, the periumbilical course of these vessels was accurately determined prior to carrying out the procedure. In our opinion, the definition of the Cruveilhier-Baumgarten syndrome needs to be extended. The results of our investigation show that, within the framework of portal neo-vascularisation, direct umbilical vein recanalisation is a relatively rare occurrence (approximately 15 per cent), while porto-femoral collateral formation is considerably more common (about 70 per cent). PMID- 2971528 TI - Source of increased serum beta-hexosaminidase in rat liver cirrhosis. AB - Lysosomal enzymes were studied by histo- and biochemical methods in cirrhotic rat livers in order to explain the serum increase of lysosomal enzymes in cirrhosis. Biochemically, the most evident increase was found in isolated hepatocytes. Histochemically, the lysosomal enzymes were increased mainly in rows of macrophages and necrotic or atrophic hepatocytes adjacent to strands of connective tissue at the periphery of the cirrhotic nodules. The results were compared to similar studies in cholestatic rats. It is concluded that most of the increased serum activities of the lysosomal enzymes derive from the hepatocytes. PMID- 2971529 TI - Biochemical studies of canine muscle phosphofructokinase deficiency. AB - Skeletal muscle from four dogs with erythrocyte phosphofructokinase (PFK; EC 2.7.1.11) deficiency were studied in vitro. Muscle PFK activities were severely decreased to 1% of the normal mean. The residual activities had a high Km for fructose-6-phosphate (F6P). Anaerobic lactate production of PFK-deficient muscle was minimal with the addition of glycogen and hexose-monophosphates, but was normal with fructose-1,6-diphosphate (FDP). Muscle glycogen concentration was twice normal, indicating a glycogen storage disorder. Histochemical studies for muscle PFK activity showed no enzymatic staining with F6P as substrate. In two muscle biopsies from asymptomatic related dogs, intermediate PFK activities were found. These data characterize canine muscle PFK deficiency in vitro. PMID- 2971530 TI - Phenotyping of phosphoglucomutase (PGM1) isoenzymes--a new method for the follow up of chimerism after bone marrow transplantation. Follow-up of chimerism after bone marrow transplantation by phenotyping of phosphoglucomutase (PGM1) isoenzymes. AB - In this study, phenotyping of phosphoglucomutase isoenzymes (PGM1, EC 2.7.5.1) in erythrocytes of bone marrow donors and hosts was shown to present a simple and exact method for the follow-up of chimerism after bone marrow transplantation, aimed at a better assessment of the transplant state and function. PMID- 2971531 TI - Inhibition of fibrinolytic enzymes by thrombin inhibitors. AB - Thrombin inhibitors have recently advanced to the stage of preclinical testing as anticoagulants. However, little is known about the effects of these inhibitors on the enzymes of the fibrinolytic system. In the present study we evaluated the effect of two protein and two synthetic inhibitors of thrombin on tissue plasminogen activator (tPA), urokinase, and plasmin. We found that hirudin inhibited the amidolytic activity of plasmin but had no effect on tPA or urokinase. Antithrombin III inhibited plasmin and urokinase but had no effect on tPA. D-Phe-Pro-Arg-CH2Cl inhibited plasmin and tPA but had no effect on urokinase. Thromstop inhibited all three fibrinolytic enzymes: plasmin, urokinase, and tPA. Thus each thrombin inhibitor tested had different inhibitory effects on the fibrinolytic enzymes. These effects should be carefully considered when thrombin inhibitors are used as antithrombotic drugs. PMID- 2971532 TI - Plasminogen activation in diabetes mellitus. Kinetics of plasmin formation with tissue plasminogen activator and plasminogen from individual diabetic donors and with in vitro glucosylated plasminogen. AB - Functional abnormalities of tissue plasminogen activator (t-PA) (high Km in the presence of fibrin) and plasminogen (Pg) (substrate inhibition in the fibrin stimulated system) from uncontrolled type I diabetics are reversible upon normalization of metabolic parameters. Therefore the effect of in vitro glucosylation of Pg was studied and similar but less pronounced substrate inhibition as with diabetic Pg was observed. However, the activation of the Pg fraction most likely containing bound cis-OH groups (e.g. glucose) was normal. These data suggest that either glucosylation in vitro is less stable than in vivo or that in diabetics protein modifications other than glucosylation occur. In this respect we can also show that acetone, a representative of ketone bodies and also elevated in poorly controlled diabetics, does not affect Pg activation in vitro. PMID- 2971533 TI - Activation of fibrin-bound plasminogen by pro-urokinase and its complementariness with that by tissue plasminogen activator. AB - Single chain urokinase (SC-UK) is a precursor of 55 kd two-chain UK (TC-UK). Treatment with catalytic proportions of plasmin or kallikrein converts SC-UK to TC-UK as a consequence of cleavage of its Lys158-Ile159 peptide bond. This plasmin-mediated activation of SC-UK induces a positive feedback secondary reaction and complicates measurement of its activity against its natural substrate, Glu-plasminogen. The fibrin-selective effect of pro-UK-induced clot lysis is not related to fibrin binding. Rather, a conformational change in Glu plasminogen, conferred when it binds to certain carboxy-terminal lysine residues on fibrin, has been implicated in this mechanism. This is complementary to t-PA. Fibrin-bound t-PA was found to exclusively activate plasminogen bound to certain internal lysine residues. Their complementariness is believed to explain their synergism in fibrinolysis. PMID- 2971534 TI - Complex rearrangements within the human J delta-C delta/J alpha-C alpha locus and aberrant recombination between J alpha segments. AB - We have examined DNA rearrangements within a 120 kb cloned region of the human T cell receptor J delta-C delta/J alpha-C alpha locus. Three types of pattern emerge from an analysis of T cell lines and clones. Firstly, cells with two rearrangements within J delta-C delta; secondly, cells with one rearrangement within J delta-C delta and one or more J alpha rearrangements, and finally, cells with rearrangements within J alpha and consequential deletion of the delta locus. Further analysis by cloning of rearrangements within the J alpha locus show that, in addition to V alpha-J alpha joins, J alpha-J alpha aberrant recombinations occur and rearrangement data indicate that such events are frequent. A model is presented to account for such recombinations. PMID- 2971537 TI - Heterogeneous distribution of calmodulin- and cAMP-dependent regulation of Ca2+ uptake in cardiac sarcoplasmic reticulum subfractions. AB - The activity of the Ca2+-pumping ATPase of cardiac sarcoplasmic reticulum is controlled by the phosphorylation level of the intrinsic membrane protein phospholamban. Phospholamban monomers contain two distinct phosphorylation sites for either the cAMP-dependent or the calmodulin-dependent kinase. The two kinases, however, preferentially phosphorylate different populations of phospholamban molecules and double phosphorylation of the same subunit by their concerted action is a phenomenon that occurs only under particular experimental conditions. This study investigates the phosphorylation pattern of phospholamban in various subfractions derived from dog cardiac sarcoplasmic reticulum. The results show that the endogenous calmodulin-dependent kinase preferentially phosphorylates the phospholamban population found in association with the cisternal compartments of the reticulum. The differential phosphorylation occurs despite the presence of sufficient amounts of the kinase in all sarcoplasmic reticulum subfractions. On the other hand, phospholamban molecules localized on the longitudinal system are preferential substrates for the cAMP-dependent kinase. Possibly, the different lipid and/or protein microenvironment of phospholamban in the various sarcoplasmic reticulum domains is responsible for the apparent heterogeneity of phosphorylation. The present findings are compatible with the concept of additive and independent action of the cAMP dependent and calmodulin-dependent kinases on cardiac sarcoplasmic reticulum. The imply, however, that different regions of the sarcoplasmic reticulum network are controlled by the two regulatory mechanisms. PMID- 2971535 TI - Thymocyte circular DNA excised from T cell receptor alpha-delta gene complex. AB - We have characterized thymocyte circular DNA excised from the T cell receptor alpha-delta gene complex. Some delta gene clones contained unusual recombinant structures derived from V-(D)-J joining: (i) a reciprocal joint of direct V to J delta joining, skipping the D delta segment; (ii) a V-D delta coding joint lacking an adjacent D delta-J delta coding joint; (iii) a V- D structure containing two D delta segments. Many of the alpha gen clones contained both coding and reciprocal joints of V alpha-to-J alpha joining on the same structure. Most of these coding joints were out of phase; however, in one clone there was an in-phase V-J alpha structure. Interestingly, some alpha gene clones contained the same V gene sequence as rearranged in the delta gene clone, indicating that the same V gene family, at least in part, could be utilized for both the alpha and delta gene systems. PMID- 2971538 TI - Cardiac failure, the kidney, renin and atrial peptides. AB - Cardiac failure is characterised by a marked reduction in renal blood flow. However, urea excretion and glomerular filtration rate remain well preserved until the terminal stages. Intrarenal compensatory actions of the renin angiotensin system probably effect this compensation. Important interrelations have recently been observed between the renin and atrial peptide systems. The latter, which is activated in cardiac failure, tends to emphasise the useful intrarenal effects while minimising the less valuable peripheral actions of the renin system in that condition. PMID- 2971536 TI - Isolation of a human gene with protein sequence similarity to human and murine int-1 and the Drosophila segment polarity mutant wingless. AB - An expressed gene sequence which was identified by the isolation of a methylation free CpG island from human chromosome 7 has been cloned from a human lung cDNA library. The deduced protein sequence contains 360 amino acids and has several features of a secreted protein; it is cysteine rich with a signal peptide sequence and two potential asn-linked glycosylation sites. The protein sequence shows marked similarity with human and murine int-1 and their Drosophila homolog wingless (Dint-1). This human int-1 related protein, int-1 and Dint-1 have diverse patterns of expression, but the inferred structural similarities suggest that some of the functional characteristics of these proteins may be shared. PMID- 2971539 TI - Atrial natriuretic factor in experimental and human hypertension. AB - Plasma levels of immunoreactive atrial natriuretic factor (IR-ANF) were evaluated by radioimmunoassay in several models of experimental hypertension and in human hypertension. Plasma levels of IR-ANF are consistently increased in all models of experimental hypertension studied so far. This is accompanied by a decrease of IR ANF levels in the left atrium at certain times. Plasma levels of IR-ANF are not increased in human essential hypertension, except in the severe form (diastolic blood pressure above 110 mmHg). Peripheral levels of IR-ANF in renovascular hypertension do not differ from normal but are increased above normal in aortic blood. PMID- 2971540 TI - Renal responses to atrial natriuretic factor in patients with congestive heart failure. AB - It is now recognised after many years of speculation regarding the role of mammalian atria in the direct regulation of intravascular volume, that atrial myocites produce a peptide hormone that results in natriuresis and diuresis, under the condition of atrial distention. This atrial natriuretic factor (ANF) is secreted in normal subjects and is increased in patients with congestive heart failure. Atrial natriuretic factor may be administered exogenously and the results of studies performed by our group indicate that natriuresis and diuresis are induced in normal subjects but not in patients with congestive heart failure. These findings, when taken together, suggest that ANF is increased in heart failure patients in response to the atrial distention associated with ventricular dysfunction and intrinsic renal insufficiency. The mechanism for the blunted renal response to atrial natriuretic factor in heart failure patients requires further clarification. PMID- 2971541 TI - Cardiovascular consequences of heart failure. AB - The cardiac and reflex neuroendocrine consequences of heart failure are reviewed. The mechanisms responsible for the many neuroendocrine reflexes that result as a consequence of heart failure are considered particularly in relation to their pathophysiological interactions. The lack of knowledge regarding quantitative changes occurring in the various circulatory reflexes activated by heart failure and the paucity of knowledge of the immediate and long-term effects of pharmacotherapeutic interventions on these reflexes are highlighted. PMID- 2971542 TI - The use of endomyocardial biopsy in heart failure. AB - Endomyocardial biopsy in this study of 1250 biopsied patients (mean of five samples/patient) proved to be a remarkably safe technique with no lethal complications. It may help to detect the underlying cause of heart failure but is handicapped by sampling error in focal disease processes (such as myocarditis and sarcoid heart disease) when conventional light and electron microscopy are used. In this biopsy series 123 patients (9.8%) suffered from severe heart failure; lymphocytic infiltrates were found in only 10 (8%). Immunohistological data suggested a secondary humoral immunopathogenesis in all patients with myocarditis and perimyocarditis, in 75% of patients with postmyocarditic heart muscle disease and in 48% of patients with primary dilated cardiomyopathy. There may thus be a need for a new classification of heart muscle diseases that includes immunological parameters of humoral and cellular autoreactivity. PMID- 2971543 TI - Humoral adaptation mechanisms and clinical outcome during vasodilator therapy. AB - Administration of hydralazine and isosorbide dinitrate leads to a reduction of one-year mortality by 38%. However, a major disappointment has been the inability to predict the individual patient's response to these drugs. The aim of the present study, therefore, was to evaluate possible predictors for a favourable response to hydralazine treatment on the basis of the acute humoral and haemodynamic response to the drug. Stroke volume increased in all 37 patients with severe left ventricular dysfunction in response to acute intravenous hydralazine (0.4 mg kg-1). However, when re-investigated after 2 months of chronic oral hydralazine therapy, sustained haemodynamic and clinical improvement was observed in only 16 patients. Pretreatment values of either left ventricular filling pressure, cardiac index or systemic vascular resistance were not useful in predicting the outcome of treatment. However, when patients were subdivided on the basis of their heart rate response to acute hydralazine, better discrimination for a beneficial response, was possible. It is concluded that the haemodynamic and humoral responses to acute intravenous hydralazine allow discrimination between patients not responding to chronic vasodilator treatment and those who are likely to benefit. A reflex excitation of sympathetic outflow in response to acute vasodilation (indicated by a reflex acceleration of heart rate and increase in plasma noradrenaline levels) characterised best those patients who did not benefit from oral hydralazine. The data indicate a detrimental role of reflex sympathetic stimulation which may blunt at least part of the direct beneficial drug-induced effect in chronic vasodilator therapy. PMID- 2971544 TI - Percutaneous transperitoneal nephrolithotomy. AB - The removal of a recurrent calculus from the pyeloureteral junction of a malrotated kidney by means of laparoscopically controlled percutaneous transperitoneal nephrolithotomy is described. PMID- 2971545 TI - Hormonal changes induced by the pure antiandrogen flutamide in postmenopausal women with advanced breast cancer. AB - Hormonal changes induced by the pure antiandrogen flutamide were studied in three postmenopausal metastatic breast cancer patients. The drug was administered at a dose of 250 mg, three times a day for 3-6 months. In each patient a sharp decrease of about 50% was observed in the circulating levels of DHT and DHEAS, irrespective of pretreatment values. A concomitant, although less pronounced, reduction in circulating testosterone, androstenedione and estradiol was found. A decrease in circulating steroids was associated with a 30% decrease in SHBG concentrations in two patients; in the third patient a 30% increase occurred. Androgens in urine, namely testosterone, androstanediol and 17-KS, decreased accordingly. In addition, a marked decrease in 17-OHCS occurred in two of the patients. These data indicate that flutamide is an effective antiandrogen in women and suggest two possible mechanisms by which the drug exerts its antiandrogenic activity: (a) inhibition of conversion of testosterone into the more active DHT, and (b) inhibition of synthesis of the adrenal precursors of active androgens. Minor variations in circulating LH and FSH were observed. Pretreatment prolactin values, which were higher than normal, dramatically decreased by 90% in one patient who had a partial remission of her disease, and they further increased in another patient who relapsed while on therapy. PMID- 2971546 TI - Effects of posture and saline infusion on atrial natriuretic peptide and haemodynamics in patients with Bartter's syndrome and healthy controls. AB - In order to explore the role of atrial natriuretic peptide (ANP) in Bartter's syndrome, five patients and five healthy controls matched for age and sex were studied. The study was designed to stimulate and suppress ANP secretion by manipulation of right atrial pressure with different body positions and mild volume expansion with saline. Other vasoactive hormones were also measured, and heart rate and blood pressure were recorded at 5-min intervals. Plasma ANP levels increased after head-down tilt and returned to baseline in the upright position. Infusion of saline failed to increase plasma ANP both in the control group and in four of the patients. No significant differences were found in plasma atrial natriuretic peptide concentrations between both groups. In view of previously reported elevated plasma ANP levels, Bartter's syndrome may be heterogeneous in this respect. Plasma renin activity was higher in the patients, but plasma aldosterone, adrenaline and noradrenaline were similar in both groups. Mean arterial blood pressure was similar in both groups, but rose significantly in the upright position in the control group only, while changes in heart rate were similar in both groups. We conclude that atrial natriuretic peptide does not seem to play a causal role in our patients with Bartter's syndrome. PMID- 2971547 TI - Decreased prostacyclin sensitivity of platelets in patients with Behcet's syndrome. AB - Patients with Behcet's syndrome have an increased risk of arterial and venous thrombosis, and abnormal platelet function has been implicated. Platelet function was studied in nine patients with Behcet's syndrome and in nine age- and sex matched healthy volunteers. Platelet aggregation in response to ADP was measured, and the threshold concentration required to produce irreversible aggregation determined. Sensitivity of platelets to the inhibitory effect of prostacyclin was also determined. In addition, plasma levels of the platelet-specific proteins, beta-thromboglobulin and platelet factor 4, and stimulated platelet thromboxane B2 production, were measured. Platelets from patients with Behcet's syndrome showed normal aggregation in response to ADP, irrespective of disease activity. Platelet sensitivity to prostacyclin was, however, decreased compared with controls--with a mean prostacyclin ID50 of 5.5 +/- 1.3 ng ml-1 (mean +/- SEM) and 1.9 +/- 0.3 ng ml-1, respectively (P less than 0.01). This reduction in platelet sensitivity to prostacyclin was greatest in patients with the most active disease. These results suggest that Behcet's syndrome may be associated with altered platelet function, and this may have important consequences with regard to the increased risk of thrombosis associated with this condition. PMID- 2971548 TI - Effect of exercise on plasma atrial natriuretic factor and cardiac function in men and women. AB - In order to provide an integrated view of the physiology of atrial natriuretic factor (ANF) during exercise, we studied changes of its plasma concentrations in 13 normal subjects (seven males, six females) during three graded exercise levels and two periods of recovery (5 and 30 min), concomitantly with an assessment of cardiac function and ventricular volumes by multigated radionuclide angiography. Mean ANF levels (+/- SEM) increased in all patients at the second (P less than 0.002) and third (P less than 0.002) exercise levels, and after 5-min recovery (P less than 0.01): in males from 16 +/- 7 to 30 +/- 11 pg ml-1 at the third level, in females from 27 +/- 12 to 61 +/- 33 pg ml-1. Normal values were observed after 30-min recovery. Even if mean ANF levels were all higher in females, this difference did not reach statistical significance (P = 0.06). Significant decreases of ventricular volumes, as well as increases of ejection fraction and rate pressure product, were noted during exercise and were similar in both sexes. The kinetics of plasma ANF concentrations, compared with the increase of rate pressure product, was characterized by a latency and a remanence in recovery. This remanence, also present in the changes of ventricular volumes, supports the hypothesis that other factor(s) like catecholamines might still exert their influence after the exercise stops. PMID- 2971549 TI - 24-hour control of blood pressure by once daily doxazosin: a multicentre double blind comparison with placebo. AB - The antihypertensive efficacy of the new, once daily, alpha 1-adrenoceptor inhibitor, doxazosin, was compared with placebo in 40 patients with mild to moderate hypertension. Following a dose titration the mean final daily doxazosin dose in 20 patients was 13.1 mg. Through-the-day blood pressure control was assessed by frequent measurements during 24 h hospitalisation in the 9 th week of double-blind treatment compared with similar measurements made during a 2 week single-blind placebo run-in. Mean reductions in standing and supine systolic and diastolic blood pressure during doxazosin treatment were statistically significantly greater than during placebo treatment at most hourly time points during the 24 h post-dose period. Twenty-four post-dose the mean falls in standing and supine diastolic blood pressure during doxazosin treatment were statistically significant when compared with placebo. Adverse effects during doxazosin treatment were generally minor and were tolerated or disappeared with continued therapy. No patients were withdrawn from the study due to adverse effects. We conclude that once daily doxazosin provides smooth and effective blood pressure control throughout a 24 h post-dose period. PMID- 2971550 TI - Lack of potentiation by cetirizine of alcohol-induced psychomotor disturbances. AB - The possible interaction between cetirizine 2 HCl and alcohol was studied in a double-blind, cross-over trial in 36 healthy volunteers. The performance of each volunteer was tested with a battery of five objective psychometric tests, and their subjective feelings were evaluated by visual analogue scales, and by extensive questionnaire. The psychomotor performance and subjective inventory of health was significantly impaired by alcohol intake. Cetirizine 2 HCl was not different from placebo. There was no adverse interaction between cetirizine 2 HCl and alcohol. Cetirizine and alcohol did not modify each others blood levels. PMID- 2971551 TI - The Fc valency of an immune complex is the decisive factor for binding to low affinity Fc gamma receptors. AB - Tetanus toxoid (TT) was complexed with two human monoclonal antibodies. The antibodies recognized different, nonrepeating epitopes. The complexes formed were characterized by gel filtration and isokinetic sucrose density gradient centrifugation. It was found that in antigenic excess the separate antibodies formed a complex of one antibody molecule and two TT molecules [IgG1-(TT)2 and IgG3-(TT)2]. In cases where equal amounts of TT and both antibodies were mixed, a dimeric complex [IgG1-(TT)2-IgG3] was formed. The binding of these immune complexes to human neutrophils and eosinophils was studied. Whereas the immune complexes containing one antibody did not bind to either cell type, the two antibody complex bound to both. This indicates that not the sterical change in the Fc part of an antibody molecule after binding an antigen, but the Fc valency of an immune complex is the decisive factor in Fc receptor interaction with neutrophilic and eosinophilic granulocytes. PMID- 2971552 TI - Stimulation-dependent lymphokine mRNA levels in human mononuclear cells. AB - Steady-state mRNA levels for lymphokines were studied in human peripheral blood mononuclear cells stimulated by various agents. The pattern of mRNA levels differed according to the way in which cells were stimulated or activated. Stimulation by anti-CD3 antibodies led to increased mRNA levels for interferon gamma but only extremely low levels of interleukin 2 mRNA were detected. In order to observe levels of interleukin 2 mRNA, it was necessary to use phorbol myristate acetate (PMA) in addition to either lectins or anti-CD3 antibodies. PMA by itself was already able to induce high levels of mRNA for granulocyte macrophage colony-stimulating factor. Most interestingly, different patterns of lymphokine production were obtained with stimuli yielding the same number of activated cells and the same ratio of activated CD4 and CD8 cells. Thus, different T cell stimuli induced distinct patterns of lymphokine mRNA levels. PMID- 2971554 TI - 5-HT1B receptors are negatively coupled with adenylate cyclase in rat substantia nigra. AB - We measured the inhibition of forskolin-stimulated adenylate cyclase by 5 hydroxytryptamine (5-HT) and other serotonin agonists in rat substantia nigra homogenates. 5-HT, 5-methoxy-3-(1,2,3,6-tetrahydro-4-pyridinyl)indole (RU 24969), 5-carboxamidotryptamine (5-CT), 1-(m-trifluoromethyl-phenyl)piperazine (TFMPP) and tryptamine inhibited forskolin-stimulated adenylate cyclase with EC50 of 67, 40, 83, 100 and 200 nM respectively. 8-Hydroxydipropylaminotetralin (8-OH-DPAT) and ipsapirone, both 5-HT1A-selective drugs, were respectively weak and ineffective to inhibit forskolin-stimulated adenylate cyclase. CGS 120 66B was almost as potent (EC50 = 100 nM) as 5-HT to inhibit the forskolin-stimulated adenylate cyclase in rat substantia nigra homogenates whereas this preferential 5 HT1B agonist was 100 times less potent than 5-HT in hippocampus guinea pig homogenates. Spiroperidol, mesulergine and ketanserin, which are potent 5-HT1A, 5 HT1C and 5-HT2 antagonists respectively, were unable to reverse the 5-HT-mediated inhibition of forskolin-stimulated adenylate cyclase whereas the beta adrenoceptor antagonists, (+/-)-cyanopindolol and (+/-)-propranolol or metergoline, fully reversed the 5-HT effect with calculated Ki of 34 +/- 18, 82 +/- 19 and 248 +/- 47 nM, respectively. The pharmacological profile of the 5-HT receptor mediating the inhibition of adenylate cyclase in substantia nigra indicates that this receptor probably corresponds to 5-HT1B binding sites. Our conclusion is that, in addition to the 5-HT1A receptor, the 5-HT1B receptor is also negatively coupled to adenylate cyclase. PMID- 2971553 TI - Phenotypic characteristics of cells involved in induced suppression to murine experimental autoimmune thyroiditis. AB - Suppression of induced experimental autoimmune thyroiditis can be consistently transferred with spleen cells to syngeneic recipients, provided they are first treated with 200 rads irradiation. Treatment with anti-Thy-1 in vivo immediately prior to transfer abrogates the suppression, while depleting B cells has no effect. The in situ induced tolerance can be prevented by treatment with monoclonal antibodies to the Ly-1 or L3T4 molecules either prior to or post tolerization. Anti-Ly-2 treatment has no effect. In the transfer, again treatment of donors with anti-L3T4 prior to transfer prevents the demonstration of suppression in the recipients, while anti-Ly-2 does not affect suppression. These data suggest that the suppression is being mediated either by a CD4+ T suppressor cell or a CD4+ suppressor inducer cell. Preliminary experiments do not support the possibility of a CD8+ suppressor cell being activated in the recipient. PMID- 2971555 TI - Cardiopulmonary effects of celiprolol and bisoprolol in serotonin-infused cats. AB - The effects of celiprolol and bisoprolol on cardiopulmonary function in serotonin infused cats were compared. Celiprolol reversed the bronchoconstrictive effect of serotonin at doses greater than or equal to 1.0 mg/kg. Also, decreases in mean arterial pressure and heart rate were noted after administration of 3-10 and 10 mg/kg celiprolol, respectively. In contrast, bisoprolol tended to induce bronchoconstriction. Reductions in mean arterial pressure and heart rate were observed with 1 or 3 mg/kg. Bisoprolol administration at 10 mg/kg was lethal. The unique ability of celiprolol to induce bronchodilation enhances its therapeutic potential. PMID- 2971557 TI - Cell-associated proteoheparan sulfate from bovine arterial smooth muscle cells. AB - Cell-associated proteoheparan sulfate has been isolated from bovine arterial smooth muscle cells preincubated with [35S]sulfate or a combination of [3H]glucosamine and [35S]methionine. The purified proteoheparan sulfate had an apparent Mr of 200,000 on calibrated Sepharose CL-2B columns. The glycosaminoglycan component (Mr approximately 30,000) was identified as heparan sulfate by its susceptibility to specific enzymatic and chemical degradation. After degradation of the proteoheparan sulfate by microbial heparitinase the resulting protein core had an apparent Mr of 92,000 on SDS-polyacrylamide gels. Its mobility was similar in the absence and presence of reducing agents indicating that the protein core consists of a single polypeptide chain. Pulse chase experiments revealed that about 40% of the cell layer-associated proteoheparan sulfate was released into the medium, while the remainder was internalized and converted to smaller species through a series of degradation steps. Initially there was a proteolytical cleavage of the protein core generating glycosaminoglycan peptide intermediates with polysaccharides chains similar in size to the original. The half-life of the native proteoheparan sulfate was found to be about 4 h. PMID- 2971556 TI - 12-O-tetradecanoylphorbol-13-acetate disrupts actin filaments and focal contacts and enhances binding of fibronectin-coated latex beads to 3T3-L1 cells. AB - The effect of a tumor-promoting phorbol ester on the binding of fibronectin coated beads to 3T3-L1 cells was studied to clarify the relationship between the binding of fibronectin to the cells, cell adhesion, and the organization of actin filaments. Interference-reflection microscopy revealed focal contacts of 3T3-L1 cells with the substratum. Stress fibers observed after rhodamine-phalloidin staining were well-developed in the cells. Treatment of the cells for 20 min with 12-O-tetradecanoylphorbol-13-acetate (TPA), but not with phorbol, disrupted focal contacts and caused a reorganization of stress fibers to generate actin ribbons. Treatment of the cells with TPA enhanced the binding of beads coated with human plasma fibronectin to the cells, as observed after incubation for 6 h with the beads. The TPA-induced increase in the percentage of cells with bound beads was dependent on the duration of treatment with TPA and on the concentration of TPA. Treatment of the cells with TPA also enhanced proliferation of cells in a dose dependent manner. The enhancement of binding of the beads by TPA was suppressed by addition of an adhesion-inhibitory peptide (Gly-Arg-Gly-Asp-Ser-Pro). Treatment with TPA did not enhance nonspecific binding of beads coated with heat denatured bovine serum albumin. Furthermore, treatment of the cells with phorbol did not enhance the binding of beads coated with fibronectin. These results suggest that TPA specifically enhances the binding of fibronectin-coated beads to 3T3-L1 cells, and that TPA-induced binding of the beads may be related to disruption of focal contacts and reorganization of actin filaments. PMID- 2971559 TI - The interaction between bone marrow immune suppression and hematopoiesis. AB - Murine bone marrow is known to contain a suppressor cell that suppresses in vitro immune responses, although its in vivo role is unknown. This cell was found to be lacking standard lymphocyte markers, including Thy 1, Lyt 1, Lyt 2, Fc receptors, and surface immunoglobin. A second cell, which acts to mask the activity of the bone marrow suppressor, was detected in neonatal mice. In the presence of this modifying cell, which was Thy 1+, the net amount of marrow suppression was decreased. A similar, though smaller, decrease in suppression could also be induced by making adult mice anemic through periodic bleeding. The parallel changes of hematopoiesis and marrow suppression suggest that these functions of the marrow are functionally linked, possibly via the Thy 1+ suppression-modifying cell. PMID- 2971558 TI - Expression of smooth muscle myosin in relation to growth kinetics of cultured aortic smooth muscle cells. AB - The goal of this study is to quantify smooth muscle myosin (SMM) expression at the level of the individual cell and to ascertain whether SMM expression in cultured aortic smooth muscle cells is related to definite growth phases, and whether the initial seeding density affects growth or SMM staining. Rabbit aortic smooth muscle cells (SMCs) were harvested by enzyme digestion of aortic tissue and plated at low (100 cells cm-2), medium (1000 cells cm-2), and high (10,000 cells cm-2) densities. Independent of seeding density, the lag phase lasted 2 to 3 days and, at all three densities, the growth rate during the logarithmic growth phase was almost the same. However, the time, the number of population doubling needed to reach the plateau phase and the cell number in the plateau were influenced by the initial seeding density. Immunofluorescence staining with anti smooth muscle myosin (ASMM) revealed intensive staining of striated and filamentous patterns in all cells during the lag and early logarithmic growth phases. During the late logarithmic growth phase, two subpopulations of cells appeared, one showing a positive and the other no reaction with SMM antiserum. The lowest relative number of cells which showed positive reactions with SMM antiserum was observed toward the end of the logarithmic growth phase. During the plateau phase, the SMM-positive subpopulation increased, amounting to about 60% of the total number of cells, independent of the seeding density. In terms of absolute numbers, the number of SMM-positive cells increased over the course of 21 days by factors of 13, 72, and 342 for high, medium, and low seeded cultures, respectively. We conclude that a SMC subpopulation can divide without loss of SMM and that some, but not all, cells which lose their SMM may possibly regain it in the postconfluent state. PMID- 2971560 TI - Trypanosoma musculi: characterization of the T-lymphocyte dependency of immunity by selective immunomodulation of the mouse, Mus musculus. AB - Trypanosoma musculi is a hemoflagellate parasite producing a characteristic course of infection in the mouse, terminating in elimination of the parasite from the vascular system with permanent immunity to reinfection. While this elimination is known to require functional T lymphocytes, the role of these cells has remained undefined. The current study examined the role of T-lymphocyte dependency by the selective in vivo depletion of T-lymphocyte subsets by monoclonal antibodies or immunosuppressive pharmacologic agents. In the former approach, monoclonal antibodies specific for the murine T-lymphocyte surface antigens Thy 1.2, Lyt 2, or L3T4 were administered to mice, resulting in functional depletion of either all T lymphocytes or the T-helper or T-cytotoxic subsets. Subsequent infection with T. musculi revealed that elimination of the T helper, but not the T-cytotoxic, phenotype resulted in a prolonged and elevated parasitemia. This enhancement of parasitemia was similar to that produced with depletion of all T lymphocytes (i.e., Thy 1.2+ cells), suggesting that the T helper cell was most important in parasite elimination. In the latter approach, pharmacologic agents displaying selective immunosuppressive activity were employed to inactivate, rather than destroy, T-lymphocyte subpopulations. In vivo inactivation of T-helper lymphocytes by the fungal metabolite cyclosporin-A enhanced the level of parasitemia in treated animals and resulted in a protracted duration of infection. In contrast, treatment with the highly substituted anthracenedione AEAD, which has been demonstrated to selectively inactivate T cytotoxic cells, had no appreciable effect on the kinetics of infection. These results strongly suggest that the T-lymphocyte dependency of Trypanosoma musculi elimination is due predominantly to the action of the T-helper, rather than T cytotoxic, lymphocyte. PMID- 2971561 TI - The effects of malnutrition on murine peritoneal macrophages. AB - Differences were detected between peritoneal macrophages (both resident and elicited) from mice on a low protein diet and from normal animals. The concentration of resident peritoneal macrophages was lower in animals on low protein diets than in normal controls. Although total protein (and therefore cell mass) of resident macrophages from malnourished mice was increased, their contents of thiamine pyrophosphatase, succinate dehydrogenase, and non-specific esterase were disproportionately reduced. In addition they did not ingest as many glutaraldehyde-fixed sheep erythrocytes or attach to as many adherent C3b sensitized sheep red blood cells as those from normal animals, although reduction of nitroblue tetrazolium was unaffected. Initially (24 hr after thioglycollate), elicited macrophages from malnourished mice did not divide as frequently as those from normal mice but by 48 hr the differences were insignificant. The elicited macrophage possessed lower levels of total protein (indicating a reduced cell mass); the levels of acid phosphatase, thiamine pyrophosphatase, succinate dehydrogenase, and nonspecific esterase and nitroblue reducing activity were also proportionately reduced. They ingested fewer glutaraldehyde-fixed erythrocytes and reacted with fewer C3b sensitised sheep red blood cells than those from normal mice; ingestion of IgG-coated sheep erythrocytes, on the other hand, was somewhat increased. These abnormalities may influence adversely the efficiency of early phlogistic responses and favor the establishment of infection in malnourished animals. PMID- 2971562 TI - Posture-induced airflow limitation in asthma: relationship to plasma catecholamines and an inhaled anticholinergic agent. AB - Postural influence on ventilatory function was investigated in thirteen asthmatic subjects on three consecutive days starting at 10.00 am. Lung function was measured in the seated position before and after four hours lying supine. Peak expiratory flow (PEF) was measured every 0.5 h in the supine position. Blood samples for determination of plasma catecholamines were collected before, during and after lying supine. After the initial lung function testing, placebo or ipratropium bromide (0.125 mg) was inhaled. On the third day the whole trial was performed seated, without any drug, as a control experiment. On the placebo day lying supine induced an initial, rapid fall of PEF followed by a progressive decrease during the four hours. The progressive decrease in PEF was apparently caused by bronchoconstriction. Ipratropium bromide prevented this posture-induced bronchoconstriction. On the day seated there was also a tendency towards a decline of PEF though less pronounced than in the supine position on the placebo day. No significant alterations in plasma levels of catecholamines were observed. We conclude that the supine posture is a stimulus to bronchoconstriction in asthma, likely to be involved in nocturnal wheezing. Postural bronchoconstriction is not explained by lowered plasma levels of adrenaline, as has been suggested for nocturnal asthma. The results raise the question of whether cholinergic mechanisms are involved. PMID- 2971563 TI - Clinical significance of abnormal rib cage-abdominal motion. PMID- 2971564 TI - Cumulative dose-response curves for assessing combined effects of salbutamol and ipratropium bromide in chronic asthma. AB - We investigated whether salbutamol (S) and ipratropium bromide (IB) exerted a true additive bronchodilator effect in asthma. In fifteen selected chronic asthmatics, individual cumulative dose-response curves to S and IB were performed on two separate days (linear regression of bronchodilator response (delta FEV1) between 20 and 80% of maximal response, versus log dose), and the dose of S equipotent to the IB dose giving the maximal bronchodilator effect (IBopt) was calculated by interpolation of each S curve. On two other days, each patient received IBopt or the equipotent S dose followed by an additional 400 micrograms S. On day 1 or 2, FEV1 reached 220 +/- 410 ml and 2410 +/- 380 ml (p less than 0.05) after the maximal dose of IB and S respectively. On day 3 or 4 after pretreatment by IB or S an additional 400 micrograms S gave a further increase, which was similar in both series (315 and 320 ml, respectively). FEV1 after combination treatment reached 238 +/- 350 ml and was not significantly different from the maximal effect of S (2440 +/- 290 ml). We conclude that S and IB exert a true pharmacological additive effect, since the combination effect is as great as the maximal effect of the most potent drug (S) and greater than the maximal effect of IB, and that the same additional dose of S gives the same increase after equipotent doses of S and IB. PMID- 2971565 TI - Effect of three different bronchodilators during an exacerbation of chronic obstructive pulmonary disease. AB - This study evaluates the effect of three different bronchodilators (beta 2 adrenergic, anticholinergic and methylxanthine) alone and in randomized sequence, during an exacerbation in thirteen patients with chronic obstructive pulmonary disease. Dose-response curves were obtained for inhaled salbutamol and inhaled ipratropium bromide. The bronchodilator effect of a perfusion of aminophylline was also assessed. When a plateau of bronchodilatation was achieved with one agent, one dose of a second bronchodilator was administered to see whether additional bronchodilation could be achieved. The increments in FEV1 and FVC were similar with the three agents. The addition of a second bronchodilator did not result in significant increments in most of the patients. In at least half of the patients the doses of salbutamol and ipratropium that produced the maximal bronchodilatation were twice that currently employed. PMID- 2971566 TI - Tuberculous pseudo-aneurysm of the descending thoracic aorta. AB - A tuberculous pseudo-aneurysm of the thoracic descending aorta was found in a 57 year-old female with haemoptysis three months after successful chemotherapy for a miliary tuberculosis. After aortic angiography, the aneurysm was resected and the aorta was repaired with a Dacron prosthetic graft. The patient is doing well one year after the operation. PMID- 2971567 TI - Cloning and sequence analysis of a cDNA encoding the beta-subunit of mouse beta hexosaminidase. AB - A cDNA encoding the prepro-beta-polypeptide of mouse beta-hexosaminidase (Hex) was isolated from a mouse lymphoblast cDNA library. The cDNA contains an open reading frame corresponding to a polypeptide of 536 amino acids which shows 74% homology with the human prepro-beta-polypeptide. An examination of the amino acid sequence identifies a putative signal peptide and five possible glycosylation sites, two of which are identical to the confirmed glycosylation sites of the human beta-chain. The amino acid sequence also shows a structurally similar though not identical site for internal cleavage responsible for the generation of mature beta a- and beta b-polypeptides. PMID- 2971568 TI - An antiserum to the C-terminus of the Alzheimer amyloid precursor recognizes a soluble 70 kDa protein. AB - A rabbit antiserum to the C-terminus of the putative brain amyloid precursor was used to probe Western blots of tissue proteins separated by SDS-PAGE. The antiserum specifically labelled a protein of approx. 70 kDa in the Tris buffer soluble fraction of brain samples from rat, Alzheimer subjects, cases of young and old Down's syndrome, and age-matched controls. The 70 kDa protein was present in low concentrations in human liver and kidney, and was undetectable in human skeletal muscle. The 70 kDa protein may be a metabolite of the amyloid precursor. PMID- 2971569 TI - NMR studies identify four intermediate states of ATPase and the ion transport cycle of sarcoplasmic reticulum Ca2+-ATPase. AB - Water proton nuclear relaxation measurements are used to detect and characterize four distinct intermediate states for Gd3+ bound to Ca2+ sites of sarcoplasmic reticulum Ca2+-ATPase in complexes with ATP analogues. In the absence of nucleotides, Gd3+ binds to two occluded Ca2+ transport sites on Ca2+-ATPase which have a low accessibility to solvent water. In the presence of the nonhydrolyzable ATP analogue, Co(NH3)4AMPPCP, a new state for bound Gd3+ (still occluded and with fewer waters of hydration) is observed. In the presence of Co(NH3)4ATP or ATP, two additional states for bound Gd3+ are detected in the NMR studies. The first of these probably represents an intermediate state for bound Gd3+ during ATP hydrolysis. The latter is the most occluded Gd3+ site yet observed in these studies and is probably analogous to the highly occluded E1-P state observed with CrATP [(1987) Biochim. Biophys. Acta 898, 313-322]. PMID- 2971570 TI - The 'cation-dependent' mannose 6-phosphate receptor binds ligands in the absence of divalent cations. AB - The requirement of divalent cations for binding of the 46 kDa mannose 6-phosphate receptors to phosphomannan and pentamannose 6-phosphate-substituted bovine serum albumin was examined. Receptors from human liver and human brain bound to both affinity ligands in the absence or presence of divalent cations with similar efficiency. The requirement for divalent cations therefore appears not to be necessary for the binding. PMID- 2971571 TI - Growth inhibitory activity of atrial natriuretic factor in rat glomerular mesangial cells. AB - Atrial natriuretic factor (ANF) binding sites, or ANF-induced cGMP accumulation, have been shown in numerous tissues that may not be intimately involved in volume homeostasis. In a series of experiments, we found that quiescent rat glomerular mesangial cells in culture could be reactivated to enter the cell cycle, and that ANF had potent inhibitory effects on proliferation of these cells. [3H]Thymidine incorporation increased by fivefold in reactivated compared to quiescent cells, and ANF in physiologic concentrations inhibited this incorporation by 48%. Thus, ANF appears to be a multifunctional peptide, consistent with a number of peptide growth factors. PMID- 2971572 TI - Aorta caldesmon inhibits actin activation of thiophosphorylated heavy meromyosin Mg2+-ATPase activity by slowing the rate of product release. AB - Activation of aorta thiophosphorylated heavy meromyosin (HMM[SP]) Mg2+-ATPase activity by aorta actin and the fraction of HMM[SP]-substrate intermediate complexes bound to actin were measured simultaneously. At 25 degrees C the Km for ATPase activation and the dissociation constant for the binding reaction were similar, irrespective of the presence or absence of tropomyosin. Aorta caldesmon (0.1 mol/mol actin) inhibited ATPase activation by 80-90% but did not alter the binding of HMM[SP]-product intermediates to actin. It is concluded that caldesmon inhibits by slowing the rate-limiting release of products from the actin HMM[SP].ADP.Pi complex. PMID- 2971573 TI - The amino acid sequences of frog heart atrial natriuretic-like peptide and mammalian ANF are closely related. AB - Despite few studies conducted in non-mammalian species, there has been a number of reports pertaining to the occurrence of a natriuretic-like substance in lower organisms. Thus, an immunoreactive substance reacting with antibodies directed against mammalian atrial natriuretic factor has previously been detected both in heart atria and ventricles of a chordate, the frog. This substance was isolated and purified from frog heart atria and its amino acid sequence established. The sequence, Ala-Pro-Arg-Ser-Ser-Asp-Cys-Phe-Gly-Ser-Arg-Ile-Asp-Arg-Ile-Gly-Ala-Gln Ser-Gly - Met-Gly-Cys-Gly-Arg-(Phe), is highly homologous to known mammalian ANF sequences. However, when aligned with the complete mammalian ANF precursor sequence at positions 121 to 151, it exhibits a single amino acid insertion at position 129 and other substitutions at positions 121, 125, 133, 135, 144, 147 and 148. Some evidence is also presented concerning the occurrence of uncleaved frog pronatriodilatin, the precursor form of ANF. This study represents the first report pertaining to the structure of a non-mammalian ANF and its precursor. PMID- 2971574 TI - Successful treatment of an infected vascular graft with gentamicin beads. AB - This short report describes the successful healing of an infected axillo bifemoral graft by the insertion of Gentamicin beads, this technique cannot be recommended for all infected grafts the majority of which will have to be removed and replaced. PMID- 2971575 TI - Analysis of complications after percutaneous transluminal angioplasty of renal artery stenoses. AB - During a five-year-period percutaneous transluminal renal angioplasty (PTRA) was attempted in 90 renal arteries with 109 stenoses and 3 occlusions in 78 patients. Complications were systematically recorded and classified as major, minor and radiological-technical. Twenty-one major complications (20.8%) including one fatality occurred as well as 17 minor, (16.8%) and 37 radiological-technical (36.6%) problems. The last group showed no clinical symptoms. The frequency of complications in our series is high compared with that in a survey of ten papers reviewing results in 675 patients. The most marked discrepancy was our high frequency of septic problems. Radiological changes are not usually reported in other series, probably because they are regarded as methodological but we considered these as potentially dangerous and important to report as they can lead to clinically relevant complications. Because of the problems reported here PTRA should only be performed in centres where complications can be properly dealt with. PMID- 2971576 TI - A non-invasive method in the assessment of surgically created arteriovenous fistulae for uraemia. AB - Subcutaneous arteriovenous fistulae are constructed regularly for haemodialysis in uraemia. However there is little available data on fistula blood flow patterns. Twenty-eight radio-cephalic fistulae constructed for 18 patients were examined over a 3-month-period using range gated pulsed Doppler ultrasound. Contralateral arms were also examined. The parameters measured were mean arterial and venous diameter, mean volume flow and mean velocity flow. The values obtained from successful and failed fistulae were compared. There was no statistical difference between either arterial or venous diameter in the two groups. Mean velocity flow and mean volume flow in the successful group however did differ significantly from those that failed. Unless the values obtained at 48 h are significantly higher than the contralateral arm, then the fistula seems unlikely to succeed. It would also seem that a simple Doppler flow meter which gives reliable mean velocity flow values is sufficient for these investigations and may even be used preoperatively, resulting in fewer re-operations. PMID- 2971577 TI - Skin blood flow: a comparison of transcutaneous oximetry and laser Doppler flowmetry. AB - In normal limbs skin blood flow (SBF) falls on standing, but in ischaemic limbs there is a characteristic blush on dependancy. We studied the effect of changing posture in a range of ischaemic limbs, using two non-invasive techniques--laser Doppler (LD) and transcutaneous oximetry (TcPO2). Warmed TcPO2 and LD probes were placed over the first metatarsal cleft and SBF recorded with the limb horizontal, dependant and elevated. Ankle to brachial pressure ratios (A:B) were also measured. Twelve patients with lower limb ischaemia, mean A:B ratio 0.5 +/- 0.21 (+/- S.D.) were compared with 21 asymptomatic controls, mean A:B ratio 1.2 +/- 0.15. In the controls, neither TcPO2 nor LD values changed significantly with limb position. Both flux and oxygen tension were significantly lower in ischaemic limbs in all positions; on lowering the ischaemic limb, flux unexpectedly rose to 23.5 +/- 17.7 V from a mean of 14 + 9.0 V (horizontal). Both TcPO2 and LD correlated well with A:B ratio in two positions: horizontal r (TcPO2) = 0.73, r(LD) = 0.57; elevated r(TcPO2) = 0.78, r(LD) = 0.68. Overall there was a highly significant correlation between LD and TcPO2 (r = 0.6, P less than 0.001, Student's test), but this was strongest in the elevated position (r = 0.87, P less than 0.001). LD and TcPO2 relate to A:B ratio and to each other, particularly in the stressed limb; in the well perfused limb, the poorer correlation suggests skin perfusion is not linearly related to tissue oxygen tension. PMID- 2971578 TI - The effect of laparoscopic multiple punch resection of the ovary on hypothalamo pituitary axis in polycystic ovary syndrome. AB - To evaluate the hormonal effect of laparoscopic ovarian punch resection in polycystic ovary syndrome (PCO), seven PCO patients were examined for pulsatile gonadotropin secretions and pituitary sensitivity before and after operation. Marked reductions of luteinizing hormone (LH) pulse amplitudes, mean LH levels, pituitary LH responsiveness, and circulating androgen levels (P less than 0.05) were observed, 3 to 4 days and 6 weeks postoperatively. LH pulse frequency, follicle-stimulating hormone (FSH), and prolactin (PRL) did not change significantly. These results are consistent with the reported hormonal effects of wedge resection and suggest that the impairment of ovarian surface acts on the hypothalamo-pituitary axis to decrease pituitary hypersensitivity and the key factor behind the resultant hormonal change lies not deep inside the ovarian stroma but in the superficial ovarian capsule. PMID- 2971579 TI - Tumor necrosis factor in peritoneal fluid of women undergoing laparoscopic surgery. AB - The level of tumor necrosis factor (TNF) in peritoneal fluid (PF-TNF) of 74 women undergoing laparoscopy was determined. The difference between the mean concentration of PF-TNF of women with normal pelvic anatomy and women with moderate or severe endometriosis was significant (P less than 0.01). The proportion of PF-TNF-positive women with PID and those with moderate or severe endometriosis was also significantly higher when compared to women with normal pelvic anatomy (P less than 0.05; P less than 0.02). The proportion of PF-TNF positive women among nulligravid and nulliparous women was significantly higher than that of women with two or more pregnancies (P less than 0.01) and two or more deliveries (P less than 0.005). These results indicate that the presence of PF-TNF is associated with primary infertility and endometriosis. PMID- 2971580 TI - [Epidermal lectin binding in psoriasis vulgaris. II. Mannosyl glycoconjugates]. PMID- 2971581 TI - [The effectiveness of prednisolone in zinc oxide containing ointments]. PMID- 2971582 TI - Efficacy and tolerance of a miconazole-benzoyl peroxide cream combination versus a benzoyl peroxide gel in the topical treatment of acne vulgaris. AB - 62-65% reduction in the number of lesions was obtained in the treatment of 52 patients with acne vulgaris in this randomized single-blind study comparing a 5% benzoyl peroxide/2% miconazole cream (BPO-MCZ) with a 5% benzoyl peroxide gel (BPO). While BPO gel was equally effective in male and female patients (66 and 73% reduction of lesions), the BPO-MCZ cream was significantly more effective in the latter (50 vs. 74% mean reduction of lesions). Tolerance was significantly better with the BPO/MCZ cream preparation, particularly in females. PMID- 2971583 TI - 15-year survey of tinea faciei in the adult. AB - In a series of 100 adults with tinea faciei various dermatologic manifestations were mimicked, viz.: discoid lupus erythematosus in 52 patients, lymphocytic infiltration in 15, seborrheic dermatitis in 11, rosacea in 8, contacts dermatitis in 7, polymorphous light eruption in 4 and granuloma faciale in 3. Trichophyton rubrum was isolated in 78% of the respective cases, other isolated organisms being Microsporum canis (13 cases), Trichophyton violaceum in 6, Trichophyton mentagrophytes (asteroides) in 2 and Epidermophyton floccosum in 1 case. In 85% of these patients the nails were also involved by the same agent found in the lesions of the face. On the basis of these observations it is recommended that all adult patients with tinea faciei should undergo a comprehensive mycological investigation to find the primary focus, which may be an infected nail. PMID- 2971584 TI - Epidermolytic palmoplantar keratoderma of Vorner: is it the most frequent type of hereditary palmoplantar keratoderma? AB - In a retrospective study, we reevaluated the biopsies that had been obtained, during the past 11 years, from 26 patients presenting with hereditary palmoplantar keratoderma (PPK). Twelve out of 26 biopsies disclosed the histological features of epidermolytic hyperkeratosis, consistent with the diagnosis of epidermolytic PPK of Vorner. A review of the histologically examined cases of the literature revealed a comparable predominance of this hereditary PPK. We conclude that, in contrast to the current opinion, epidermolytic PPK of Vorner represents the most frequent type of hereditary PPK. PMID- 2971585 TI - Perinatal correlates of major and minor neurological dysfunction at school age: a multivariate analysis. AB - A prospective study was carried out on 747 infants: 147 neurologically abnormal, 300 with mild neurological abnormalities and 300 normal infants. They were re examined at nine years of age, with special attention being paid to minor neurological dysfunction (MND). Extensive data on obstetrical history, neonatal course and interval complications were collected. Neurological handicap was not found on follow-up in the group of normal newborns. Other than neonatal neurological abnormality, risk factors contributing significantly to later handicap were low one-minute Apgar scores, a disturbed neonatal course, low social-class and interval complications; obstetrical events were conspicuous by their absence. Two aetiologically and clinically distinct kinds of MND were distinguished on the basis of a neurological cluster profile: MND-1 (one or two abnormal clusters) was only associated with a birthweight below the 2.3 centile and male gender, and MND-2 (more than two abnormal clusters) was associated with neonatal neurological findings, social class, obstetrical optimality score and gender. PMID- 2971586 TI - Facial plastic surgery for children with Down syndrome. PMID- 2971587 TI - [Relations between polycystic ovary and idiopathic hirsutism]. PMID- 2971588 TI - [Extreme cardiac hypertrophy in athletes. Morphological and functional echographic study]. AB - Cardiac hypertrophy is an adaptation phenomenon of the heart as a result of increased hemodynamic load due to intense and prolonged training in athletes. This is mainly seen in endurance athletes. In some cases cardiac hypertrophy can mimic hypertrophic cardiomyopathy, specially if hypertrophy is mainly localized at the interventricular septum as compared to the left ventricular free wall. In our study we tried to evaluate the different diagnostic features obtained by echocardiography and clinical examination in a group of 23 athletes with marked hypertrophy (all were participants to the 1984 Olympic Games held in Los Angeles) as compared to a group of 11 sportsmen with non-obstructive hypertrophic cardiomyopathy (HC). Cardiomyopathy was diagnosed on the basis of clinical, echocardiographic and angio-scintigraphic findings. The 23 athletes were selected on the basis of M-Mode and 2D echocardiographic thickness of the interventricular septum (IVS) which was in diastole greater than or equal to 15 mm. They were all asymptomatic, only 4 of the 23 athletes had ECG anomalies due to left axis deviation (LAS) and T wave inversion. Only 4 of the subjects with HC had a family history of HC. ECG changes were the following: T wave inversion (9 subjects), left axis deviation (LAD) (4 subjects), deep Q wave in D2-3, aVF, V5-6 (2 subjects) and low voltage R wave in V5-6 (1 subject). All the athletes had marked hypertrophy of the IVS. Interventricular septum thickness (IVST) was 15.7 +/- 0.6 mm, with a range from 15 to 17.5 mm. Posterior wall thickness (PWT) was 13.7 +/- 1.1 with a range from 12 to 16 mm. The sportsmen with HC had an IVST of 16.2 +/- 3.5 mm and a PWT of 11.3 +/- 1.5 mm. The IVST/PWT ratio was significantly lower (p less than 0.01) in the athletes (1.14 +/- 0.02) when compared to the group with HC (1.4 +/- 0.3). In 7 of the subjects with HC the hypertrophy was mainly localized at the anterolateral segment of the interventricular septum (in 4 of these subjects the hypertrophy involved also the antero-lateral part of the left ventricular free wall).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2971589 TI - [Ventricular hypertrophy in athletes]. PMID- 2971590 TI - [The significance of trace elements in the development of iron-deficiency anemia in the Volga River basin]. PMID- 2971591 TI - HLA and Down syndrome (DS): parents at the origin of the nondisjunction share no more HLA-A and -B antigens with their DS child than controls. AB - Fifty couples and their children with Down syndrome (D.S.) were typed for HLA-A and HLA-B antigens and compared to 50 control families and 464 blood donors. The parental origin of the extra chromosome 21 was determined by cytogenetic methods. All individuals were caucasians and there was no history of consanguinity. No excessive HLA sharing was present in D.S. parents. The mothers of D.S. shared no more HLA antigens with their D.S. children than the control mothers with their normal children (14% vs. 18%). Thirteen of the fifty pairs (26%) (parent in whom the nondisjunction occurred and D.S. child) shared three HLA antigens at the A and/or B locus. This was not significantly higher than the proportion in the control group (12/50 or 24%). These data suggest that it is not the sharing of HLA-A and HLA-B antigens between the parents or between the parent who was the origin of the nondisjunction and the D.S. child that is related either to the occurrence of trisomy 21 zygotes or to prenatal survival of affected embryos and fetuses. PMID- 2971592 TI - Cloning of human lysozyme gene and expression in the yeast Saccharomyces cerevisiae. AB - cDNA clones encoding human lysozyme were isolated from a human histiocytic cell line (U-937) and a human placenta cDNA library. The clones, ranging in size from 0.5 to 0.75 kb, were identified by direct hybridization with synthetic oligodeoxynucleotides. The nucleotide sequence coding for the entire protein was determined. The derived amino acid sequence has 100% homology with the published amino acid (aa) sequence; the leader sequence codes for 18 aa. Expression and secretion of human lysozyme in Saccharomyces cerevisiae was achieved by placing the cloned cDNA under the control of a yeast gene promoter (ADH1) and the alpha factor peptide leader sequence. PMID- 2971593 TI - New cloning vectors and techniques for easy and rapid restriction mapping. AB - We have modified plasmid, phage lambda and cosmid cloning vectors to be of general use for easily and unambiguously determining restriction maps of recombinant DNA molecules. Each vector is constructed so that it contains the rarely found NotI restriction site joined to a short synthetic linker sequence that is followed by a multiple cloning site. DNA cloned into these vectors may be restriction-mapped by either of two methods. In one technique, the cloned DNA is completely digested with NotI, followed by partial digestion with any other restriction enzyme. After electrophoresis and transfer to a nylon membrane, the fragments are hybridized to a labeled probe complementary to the NotI linker. In the second technique, referred to as recession hybridization detection, cloned DNA is digested with NotI and then briefly treated with exonuclease III to recess the 3' ends. After hybridizing a labeled complementary oligodeoxynucleotide to the single-stranded 5' end containing the linker sequence, the DNA is partially digested with another restriction enzyme, electrophoresed and the gel is exposed to x-ray film. With either method the size of each labeled fragment corresponds directly to the distance that a restriction site is located from the NotI linker terminus. Methods for obtaining partial restriction enzyme digests have been devised so that as many as 20 different enzymes may be conveniently mapped on a single gel in little more than a day. The vectors and techniques described may also be adapted to automated or semi-automated devices that read fragment lengths and calculate the resulting restriction map.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971594 TI - Characterization and cloning of gene 5 of Bacillus subtilis phage phi 29. AB - Sequencing of the phi 29 DNA region [open reading frames (ORFs) 12, 11 and 10] between genes 6 and 4 of the mutant ts5(219) showed that a G in the wild-type phage had been changed to an A in the mutant at position 218 of ORF 10 indicating that this ORF corresponds to gene 5. ORF 10 was cloned in plasmid pPLc28 under the control of the PL promoter of phage lambda and, after heat induction of the Escherichia coli cells carrying the recombinant plasmid pGM26, a 12-kDa protein was overproduced, accounting for about 5% of the de novo synthesized protein. Introduction of a nonsense mutation in ORF 10 indicated that the latter codes for the 12-kDa protein. The predicted secondary structure, the hydrophilicity values and the antigenic regions of protein p5 are discussed. PMID- 2971595 TI - Transcriptional regulation and gene arrangement of Escherichia coli, Citrobacter freundii and Salmonella typhimurium biotin operons. AB - The bio operons of Citrobacter freundii and Escherichia coli K-12 (strain C600) were isolated by screening lambda banks for complementation of E. coli bio mutants. These were compared with the previously isolated bio operon of Salmonella typhimurium and previous data on E. coli K-12. The restriction maps of the operon are very different in the three species, but no difference in gene order was found. Operator-promoter DNA, identified by repressible titration and by biotin-repressible transcription in E. coli, was sequenced and compared to the published E. coli K-12 sequence. In the segment previously identified as operator/bioB promoter, C. freundii and S. typhimurium DNA are identical and differ from E. coli only by 2 bp. The DNA to the right of this segment (indicated by previous data to be the bioA promoter of E. coli) has diverged in all three species, and only E. coli has a sequence resembling a consensus promoter. PMID- 2971596 TI - The elderly mystique: constraints on the autonomy of the elderly with disabilities. PMID- 2971598 TI - Deficiency of autologous mixed lymphocyte reaction in long-term bone marrow transplant recipients without clinical chronic graft versus host disease. PMID- 2971599 TI - Effect of vitamins on Trichinella spiralis Owen, 1835 infection in mice. AB - The effects of vitamins A, B complex, E, and ADE on the body weight, eosinophilia, intensity of infection and distribution of T. spiralis larvae were studied in mice. The greatest loss of weight followed after the application of vitamins B complex and E. An increased eosinophilia appeared in the majority of infected mice since day 7 p.i., reaching the maximum on day 21 p.i. In mice receiving vitamins B complex, A, and ADE, the increased eosinophilia was observed still on day 60 p.i. The highest levels of eosinophilia occurred after the application of vitamins B complex and E, which was directly proportional to the intensity of infection. The lowest intensity of infection was recorded in mice receiving vitamin A. Though there were great differences between individual mice, the greatest number of larvae were localized in the diaphragm and left masseter. PMID- 2971597 TI - Efficacy of sequential cyclical hormonal therapy in endometrial cancer and its correlation with steroid hormone receptor status. AB - Forty-six eligible women with metastatic endometrial cancer were randomly allocated to receive monthly cycles of either CAF (cyclophosphamide, adriamycin, 5-fluorouracil) or CAF plus Provera 200 mg daily for 3 weeks followed cyclically by Tamoxifen 20 mg daily for 3 weeks. Overall response rates of 15 and 43% were seen with CAF and CAF plus hormonal therapy. Using a multivariate analysis of the results, this difference is significant (P value 0.05). In 8 patients with operable endometrial cancer, negative estrogen receptor concentration (ER less than 15 fmole/mg protein) and Grade 3 disease, the clinical course was aggressive in 4 patients with systemic and local relapse. In 10 other similar patients (negative ER and Grade 3) who received adjuvant cyclical hormonal therapy only 1 relapsed and the other 9 are disease-free for an average of more than 31 months. Sequential cyclical hormonal therapy with ER and progesterone receptor analysis has a place in the management of endometrial carcinoma. PMID- 2971600 TI - [Acne: long-term therapy with side effect. Sebaceous gland activity is androgen dependent--acne therapeutic drug is effective as a contraceptive]. PMID- 2971601 TI - Receptors for insulin-like growth factors in the central nervous system: structure and function. AB - Insulin-like growth factors (IGFs) I and II are homologous peptides, which stimulate growth of several vertebrate tissues. Expression of IGF I and IGF II genes and production of IGFs have recently been demonstrated in rat and human brain. In search for the function of IGF I and IGF II in the central nervous system, we have studied IGF receptors in fetal and adult mammalian brain and growth effects of IGFs on primary cultures of fetal rat astrocytes. Two types of IGF receptor are present on adult rat brain cortical plasma membranes, on fetal rat astrocytes and on human glioma cells. Type I IGF receptor is composed of 2 types of subunits: alpha-subunits which bind IGF I and IGF II with high affinity and insulin weakly, and beta-subunits which show tyrosine kinase activity and autophosphorylation stimulated by IGF I and IGF II with almost similar potency. The molecular size of the type I IGF receptor alpha-subunit is larger in cultured fetal rat astrocytes and human glioma cells than in normal adult brain (Mr 130,000 versus 115,000), whereas the beta-subunit has the same electrophoretic mobility (Mr 94,000). The type II IGF receptor is a monomeric protein (Mr 250,000), which binds IGF II 5 times better than IGF I, and does not recognize insulin. The amounts of type II IGF receptor are significantly higher in fetal and malignant cells than in adult brain. Based on these findings we suggest that IGF receptors in brain undergo changes during fetal development and malignant transformation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971602 TI - Alzheimer's disease and Down's syndrome. AB - The neuropathology of Down's syndrome at middle age is compared with that of Alzheimer's disease at that age, through a review of the published literature and from the author's personal observations. The pathological changes of Down's syndrome at middle age, i.e. the form and distribution of senile plaques and neurofibrillary tangles, and the pattern of involvement (atrophy) of neuronal systems are qualitatively the same as those of Alzheimer's disease at that age. Quantitative differences do occur and these may relate to biological or sociological variations inherent to the two parent populations. It is concluded that, in pathological terms, patients with Down's syndrome at middle age do indeed have Alzheimer's disease. Some ways in which a study of patients with Down's syndrome can give insight into the nature and development of the pathological changes of Alzheimer's disease are put forward and discussed. PMID- 2971603 TI - Single dose oral norfloxacin or intramuscular spectinomycin to treat gonorrhoea (PPNG and non-PPNG infections): analysis of efficacy and patient preference. AB - Norfloxacin, a new oral quinolone, was compared with intramuscular spectinomycin for treating culture proved gonorrhoea (caused by penicillinase producing strains of Neisseria gonorrhoeae (PPNG) and non-PPNG strains. A total of 547 infected men and women were randomly allocated to treatment with either single dose norfloxacin (800 mg by mouth) or spectinomycin (2 g intramuscularly). Patient preference for tablets or injections was noted at this visit. Patients returned four to eight days later for assessment of efficacy, safety, and preference. Of the 482 patients who attended follow up, all those treated with norfloxacin (94 infected with PPNG strains, 145 with non-PPNG strains) and all 82 infected with PPNG strains and treated with spectinomycin were cured. Of 161 infected with non PPNG strains and treated with spectinomycin, 159 (99%) were cured. Side effects (headache, nausea, and sleepiness) occurred in three patients receiving norfloxacin and in 17 (16 pain at injection site, 1 giddiness) receiving spectinomycin. Most patients preferred tablets to injection both on day 1 (313 v 200) and at follow up (373 v 104). This study showed that norfloxacin was a highly effective alternative to spectinomycin, produced fewer side effects, and was the preferred mode of administration. PMID- 2971604 TI - A comparative study of the change in ATPase activity of proteolytic fragments of myosin in relation to the presence of DTNB light chains. PMID- 2971605 TI - Expression of the VEP13 antigen (CD16) on native human alveolar macrophages and cultured blood monocytes. AB - Human alveolar macrophages (AM phi) from thirteen patients, who were suffering from various lung diseases were harvested by bronchoalveolar lavage. Peripheral blood monocytes from eight healthy donors were isolated by Ficoll-Hypaque gradient centrifugation and adherence to plastic surface. To detect the VEP13 antigen (CD16) on these cells, a rosette assay employing ox erythrocytes coated by the CrCl3 method with purified VEP13 monoclonal antibody (Eo-VEP13) was used. A mean of 31.3% of freshly isolated AM phi and 3.9% of blood monocytes formed Eo VEP13 rosettes. Monocytes cultured for 3 or 6 days in the presence of a supernatant from mouse L929 cells, which had been shown previously to improve long-term viability of human monocytes in culture, showed 12.5% and 25.3% Eo VEP13 rosettes, respectively. No significant increase in VEP13 antigen expression was noted by culturing monocytes without L929 cell supernatant. The factor in L929 supernatant that induces VEP13 antigen expression has not been identified. Tunicamycin at 10 micrograms/ml inhibited significantly VEP13 antigen expression on monocytes. In contrast, IgG rosette formation was not reduced by tunicamycin. Our data show that subpopulations of native human AM phi and peripheral blood monocytes cultured in presence of a supernatant of L929 fibroblasts containing mainly murine CSF may express the CD16 antigen, which is normally found on large granular lymphocytes (LGL). Suppression by tunicamycin indicates that Fc receptor glycosylation takes place during a later differentiation step of mononuclear phagocytes. PMID- 2971606 TI - Traffic and proliferative responses of recirculating lymphocytes in fetal calves. AB - The thoracic duct or efferent prescapular duct was cannulated in four fetal calves aged 121-259 days post-conception. The duration of lymph flow ranged from 2 to 20 days and the mean flow rates sustained over these collection periods varied from 5.4 to 48.8 ml/hr. Lymphocyte output ranged from 4.4 x 10(6) cells/hr in thoracic duct lymph from a 121-day fetus to 3.9 x 10(8) cells/hr in efferent prescapular lymph from a 259-day fetus. The circulating lymphocyte pool in fetal calves of about 120 and 190 days gestational age was calculated to contain, respectively, 4 x 10(8) cells and 2 x 10(10) cells. The proportion of lymphocytes bearing surface immunoglobulin detected in fetal lymph ranged from 2.1% to 8.7%. Recirculating lymphocytes from fetal calves produced strong proliferative responses when stimulated by T-cell mitogens but responded poorly to B-cell mitogens. Fetal lymphocytes also responded to stimulation by allogeneic cells and stimulated other cells to proliferate during mixed lymphocyte culture. When stimulated with Con A, fetal lymphocytes secreted IL-2 to a degree that was indistinguishable from the secretory behaviour of lymphocytes from adult animals. The results presented in this paper show that chronic lymphatic fistulae can be established successfully in fetal calves to give access to recirculating lymphocytes. This provides a new experimental approach for studying the development of the bovine immune system. PMID- 2971607 TI - Induction of suppressor cells by staphylococcal enterotoxin B: identification of a suppressor cell circuit in the generation of suppressor-effector cells. AB - We have shown previously that staphylococcal enterotoxin B (SEB) has the capacity to non-specifically inhibit antibody responses in vitro through the induction of a suppressor cell population. In the present studies, an analysis of the cellular dynamics has shown that the generation of Lyt-1-2+ suppressor-effector cells is dependent on the initial activation by SEB of an Lyt-1+2- suppressor-inducer population. Co-culture experiments carried out in vitro suggest that the induction of the suppressor-effector population requires at least two signals: one signal is provided by the suppressor-inducer population, and the second signal is provided by SEB. Studies also show that the in vitro antibody response is suppressed when the suppressor cells are added as late as Day 4 of a 5-day culture. While the suppressor cell population activated early in the antibody response is Lyt-1-2+, depletion studies suggest that the population that acts late in an ongoing response bears the Lyt-1+2+ surface phenotype. The results demonstrate that at least three distinct SEB-induced T-cell populations are capable of participating in the suppression of the antibody response. The relationship between the generation of non-specific suppressor cells and the activation of antigen-specific cell circuits is discussed. PMID- 2971608 TI - Genetic and stimulator cell requirements for generation and activation of minor histocompatibility antigen-specific memory cytotoxic T-lymphocyte precursors. AB - By adding IL-2 (supernatant of culture of concanavalin A-activated rat spleen cells) on Day 3 of mixed leucocyte cultures (MLC) we managed to fully activate multiple minor histocompatibility antigen (MIHA)-specific cytotoxic T-lymphocyte precursors (CTLp). In this newly developed system we studied genetic and stimulator cell requirements for the generation and activation of MIHA-specific memory CTLp. Memory CTLp were activated to generate effector CTL in MLC only when major histocompatibility complex (MHC)-compatible MIHA-allogeneic cells were used as stimulators. In contrast, memory CTLp were generated in mice that were primed by injection of either MHC-compatible or incompatible MIHA-allogeneic spleen cells. A surprisingly small number (10(4] of MHC-disparate cells cross-primed mice effectively. For priming, no special accessory cell types were required as stimulators, and 10(4) adherent cell-depleted spleen cells primed mice as well. These results contrasted to another finding that sonication-disrupted 10(6) stimulator cells did not prime mice effectively, and antigens shed from 10(7) live stimulator cells failed to sensitize host antigen-presenting cells for priming. It is suggested from these results that the mode of recognition of MIHA by virgin CTLp is unique or that an as yet unknown unusual stimulation pathway works for the priming. PMID- 2971609 TI - AK cells were developed from NK cells during in vitro culture of allogeneic or F1 anti-parental stimulation: functional conversion in recognizing H-2 expression of target cells accompanied by phenotypical conversion. AB - In vitro sensitization of (CBA x A)F1 spleen cells for 3 days with allogeneic C57BL cells raised the killer activity to the NK-sensitive YAC-1 target. When (A x C57BL)F1 spleen cells were cultured with parental C57BL cells, the lytic activity to YAC-1, P815 and EL-4 targets occurred on Day 6 after the culture. Phenotypical analyses showed that these culture-activated killer (AK) cells were derived from asialo-GM1+Thy-1-NK cells; however, they expressed Thy-1 antigen but not asialo-GM1 antigen at the effector cell level. Generation of the AK cells was not evident in cultures of spleen cells from mice with a neonatally induced tolerance to stimulator antigen and in those from T-cell-depleted mice. The supernatant of allostimulated culture, which contained a low concentration of IL 2, rendered the above cells capable of evoking AK activity. The H-2-reduced target cells were sensitive to NK cells, but less sensitive to AK cells; on the contrary, the H-2 highly expressed cells (interferon-treated cells) were less susceptible to NK cells but highly susceptible to AK cells. Thus, the relation between NK susceptibility and susceptibility to AK cells is inverse. Our study shows that stimulation with lymphokines causes a functional conversion accompanied by a phenotypical conversion of NK cells. With reference to immunosurveillance, these observations lead to the idea that NK and AK cells represent two functionally distinct but complementary systems involved in cell mediated immunosurveillance. PMID- 2971610 TI - Protein malnutrition reduces the IgA immune response to oral antigen by altering B-cell and suppressor T-cell functions. AB - Mice fed a protein-deficient diet containing 2% ovalbumin for 6 weeks (PD) show a reduced spleen IgA plaque-forming cell (PFC) response after oral immunization with sheep red blood cells (SRBC), compared to the response of mice fed a control diet containing 20% ovalbumin (C). This reduced IgA PFC response was partially restored by reconstituting the PD mice with an i.v. injection of 5 X 10(5) Peyer's patch (PP) cells from C mice prior to immunization. Reconstitution with B cell enriched PP, but not helper T-cell (Th) enriched PP, from C mice resulted in an equally elevated IgA response, suggesting that malnutrition induced alterations in the PP B cells yet allowed partially functional Th cells. Reconstitution of PD mice with 1 X 10(7) PP cells from C mice yielded a very low IgA PFC response. This suppression was relieved by removal of Lyt-2+ T cells from the control PP cells prior to transfer. Transfer of spleen, but not PP, cells from oral-immunized PD donor mice to C mice prior to oral immunization resulted in a suppressed IgA PFC response of the recipient mice that was not seen when immune C cells were transferred. Removal of Lyt-2+ cells from the PD spleen cells prior to transfer abolished this suppression. These results suggest that protein deprivation is accompanied by the stimulation of Lyt-2+ suppressor T cells that suppress the IgA response after oral immunization. PMID- 2971611 TI - Opsonin-dependent and independent surface phagocytosis of S. aureus proceeds independently of complement and complement receptors. AB - We examined the mechanism of surface phagocytosis of Staphylococcus aureus by human polymorphonuclear leucocytes (PMN). Surface phagocytosis of unopsonized bacteria occurred, but was significantly enhanced by the presence of serum. The serum requirement was low, and a maximal effect occurred with serum concentrations of 0.25-0.5%. The opsonic effect of serum was not removed by heat inactivation of complement but was adsorbed, at low serum concentrations, by protein A, indicating that opsonin-dependent surface phagocytosis requires IgG but not C3. The requirement of opsonin-dependent surface phagocytosis for IgG was demonstrated further with purified IgG preparations as the sole opsonin. Activation of PMN by N-formyl-methionyl-leucyl-phenylalanine (FMLP) or phorbol myristate acetate (PMA) increased opsonin-independent surface phagocytosis by 47% and 66%, respectively, but had no effect on opsonin-dependent surface phagocytosis. Blockade of the PMN iC3b receptor (CR3), which has lectin-like properties, by a panel of monoclonal antibodies against the alpha- and beta chains of CR3 did not inhibit the surface phagocytosis of opsonized or unopsonized S. aureus, and one antibody (NIMP-R10) enhanced opsonin-independent surface phagocytosis. These results indicate that the mechanism of surface phagocytosis is quite different to that observed in suspension assays. Opsonin independent surface phagocytosis occurs and is enhanced by PMN activation, opsonin-dependent surface phagocytosis is dependent on IgG and not complement, and neither opsonin-independent nor -dependent surface phagocytosis proceeds through CR3. PMID- 2971612 TI - Abnormal T-cell activation in chronic hepatitis B viral infection: a consequence of monocyte dysfunction? AB - The process of T-cell activation in chronic hepatitis B virus (HBV) carriers has been investigated by measurement of membrane expression of lymphocyte-activation markers in response to a variety of mitogenic stimuli in order to delineate further the abnormality of T-cell-mediated immunity present in such patients. A substantial proportion of unstimulated T cells from the peripheral blood of patients but not controls expressed HLA-DR; in contrast the IL-2 and transferrin receptors were rarely expressed spontaneously in either group and there was no difference in spontaneous lymphocyte transformation. After stimulation with monocyte-dependent T-cell mitogens, phytohaemagglutinin (PHA) or anti-T3, patients had significantly reduced expression of the IL-2 and transferrin receptors and of HLA-DR in association with impaired lymphocyte transformations compared to controls. In contrast, lymphocyte activation was normal in response to the monocyte-independent T-cell mitogen phorbol-myristate-acetate (PMA). These data confirm that the process of T-cell activation is abnormal in chronic HBV carriers but suggest that the T cell is intrinsically normal. In allogeneic co cultures, monocytes from patients inhibited the transformation of normal and patients' lymphocytes in response to PHA, suggesting that defects of T-cell mediated immunity in chronic HBV carriers may be a consequence of monocyte dysfunction. PMID- 2971613 TI - Recombinant interleukin 2 inhibits pokeweed mitogen-induced proliferation of human adult peripheral blood and cord blood mononuclear cells. AB - We studied the effects of recombinant interleukin 2 (rIL-2) on pokeweed mitogen (PWM)-induced proliferation of unfractionated human mononuclear cells from the peripheral blood (PBMCs) and cord blood (CBMCs). rIL-2 was found to inhibit PWM induced proliferation of both PBMCs and CBMCs in a dose-dependent manner. The response was statistically significant at concentration 10 U/ml. The inhibition of adult PBMC proliferation was relatively mild, but in newborns up to 75% inhibition was found. The inhibitory effect of rIL-2 was best detectable at day 4, at the same time as PWM-induced proliferation peaked. However, also at days 8 and 10, the response induced by PWM plus rIL-2 was much lower than that induced by rIL-2 alone. These finding suggest that the inhibition was due to rIL-2 mediated stimulation of the function of PWM-activated suppressor cells. CD8+ T cells were, however, not alone responsible for the rIL-2-induced inhibition of PWM-induced proliferation, since the responses were essentially similar independent of whether CD8+ T cells were present or absent. In addition, the release of interferon-gamma (IFN-gamma) in response to rIL-2 was not a major cause of the inhibitory effects, because rIL-2 inhibited PWM-induced proliferation of CBMCs, which are deficient in producing IFN-gamma. Our results provide novel data to the ongoing discussion of IL-2 as a down-regulator of immune functions. PMID- 2971615 TI - Assignment of the gene coding for human FcRII (CD32) to bands q23q24 on chromosome 1. PMID- 2971614 TI - Molecular organization of the human CD3 gene family on chromosome 11q23. AB - The genes encoding three invariant components of the human T-cell antigen receptor, the CD3 delta, gamma, and epsilon chains, are located on human chromosome 11 at band q23. We isolated cosmid clones containing the human CD3 delta and gamma chain genes in vectors designed for rapid and efficient chromosome "walking". The human CD3 epsilon gene was located in the region immediately downstream of the CD3 delta and gamma genes using synthetic oligonucleotide probes and the localization of this gene confirmed by DNA sequencing. Detailed restriction mapping of the CD3 locus demonstrated that all three CD3 subunits are encoded within 60 kb of DNA with the CD3 epsilon gene located 26 kb downstream of the CD3 delta and gamma genes. Analysis of genomic DNA on pulsed field gels using probes isolated from these cosmid clones defined a physical map of 750 kb spanning the CD3 locus on human chromosome 11q23. The CD3 genes thus comprise a multigene family encoding cell surface components important for transmembrane signaling on T lymphocytes. The arrangement of these genes suggest that they may share common regulatory elements for the control of gene expression during T-cell ontogeny. PMID- 2971616 TI - Cardiopulmonary reflex before and after regression of left ventricular hypertrophy in essential hypertension. AB - Studies that have examined the cardiopulmonary receptor control of circulation in hypertension have produced conflicting results. In 10 normotensive subjects and in age-matched essential hypertensive subjects without (n = 10) or with left ventricular hypertrophy (n = 12), as well as in seven subjects of the latter group restudied after 1 year of treatment that induced regression of cardiac hypertrophy, we examined the cardiopulmonary reflex by increasing central venous pressure and stimulating cardiopulmonary receptors through passive leg raising and by reducing central venous pressure and deactivating cardiopulmonary receptors through nonhypotensive lower body negative pressure. Reflex responses were measured as changes in forearm vascular resistance (mean blood pressure divided by plethysmographically measured blood flow), plasma norepinephrine concentration, and plasma renin activity. In hypertensive subjects without left ventricular hypertrophy, stimulation and deactivation of cardiopulmonary receptors caused changes in forearm vascular resistance, norepinephrine concentration, and plasma renin activity that were modestly reduced as compared with those in normotensive subjects. However, all these changes were markedly reduced in hypertensive subjects with left ventricular hypertrophy. Following regression of left ventricular hypertrophy, the changes in vascular resistance, plasma norepinephrine, and plasma renin activity induced by cardiopulmonary receptor manipulation all improved markedly. These results demonstrate that cardiopulmonary receptor regulation of peripheral vascular resistance and of neurohumoral variables is impaired in essential hypertension and that the impairment is much more pronounced when this condition is associated with cardiac structural alterations. Therapeutic regression of these alterations, however, leads to a marked improvement of this reflex, with consequent favorable effects on circulatory homeostasis. PMID- 2971617 TI - Role of prostaglandins in mediating the renal effects of atrial natriuretic factor. AB - The natriuretic response to the intrarenal administration of atrial natriuretic factor (ANF) is accompanied by an increase in the synthesis of prostaglandins and by a redistribution of renal blood flow from the superficial to the deep cortex. This study was undertaken to define whether prostaglandins mediate the ANF induced redistribution of renal blood flow and if prostaglandins and renal blood flow redistribution contribute to the natriuretic actions of ANF. In anesthetized dogs, the intrarenal administration of indomethacin (10 micrograms/kg/min) or the intravenous administration of meclofenamate (5 mg/kg) completely prevented the sixfold and twofold increments in urinary prostaglandin E2 and 6-keto prostaglandin F1 alpha excretion, respectively; it also abolished the redistribution of renal blood flow to the deep cortex. However, ANF induced a similar natriuresis before (from 53 +/- 17 to 281 +/- 48 microEq/min) and after (from 45 +/- 13 to 273 +/- 60 microEq/min) the administration of prostaglandin synthesis inhibitors. It is concluded that the ANF-induced redistribution of renal blood flow to the deep cortex is prostaglandin-mediated but that neither redistribution nor increased prostaglandin synthesis is an important mediator of ANF's natriuretic action. PMID- 2971618 TI - Enhanced natriuretic effect of atrial natriuretic factor during mineralocorticoid escape in humans. AB - We examined the question of whether escape from the sodium-retaining effect of mineralocorticoid involves an increased natriuretic effect of atrial natriuretic factor (ANF). Seven healthy volunteers taking a 170 mmol Na/100 mmol K diet received an intravenous bolus (25 micrograms) followed by a 1-hour infusion (0.02 micrograms/kg/min) of ANF (human ANF-[99-126]) before and after 10 days of 9 fludrocortisone acetate, 0.5 mg b.i.d. Escape was accompanied by an increase in body weight (from 72.2 +/- 12.9 to 74.0 +/- 12.6 kg; p less than 0.05), mean arterial pressure (from 95 +/- 4 to 109 +/- 3 mm Hg; p less than 0.01), plasma ANF (from 9 +/- 2 to 24 +/- 4 pmol/L; p less than 0.01), and inulin clearance (from 124 +/- 9 to 137 +/- 7 ml/min; p less than 0.05). Indexes for renal sodium handling (lithium and free water clearance) were compatible with a decreased "proximal" and an increased "distal" tubular reabsorption fraction. ANF infusion raised inulin clearance comparably before and after escape to 138 +/- 10 and 152 +/- 7 ml/min, respectively, but the natriuretic effect was much larger (p less than 0.05) after escape (from 366 +/- 34 to 1294 +/- 278 mumol/min) than before (from 248 +/- 48 to 630 +/- 124 mumol/min). Indexes for tubular reabsorption were consistent with greater suppression of both "proximal" and "distal" tubular sodium reabsorption by ANF after versus before mineralocorticoid expansion. These results indicate that escape is accompanied not only by a rise in plasma ANF but also by potentiation of the natriuretic effect of ANF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971619 TI - Effects of incremental infusions of atrial natriuretic factor on aldosterone, renin, and blood pressure in humans. AB - To evaluate the physiological effects of human atrial natriuretic factor-(99-126) (ANF), we infused ANF, 0.1, 0.3, and 1.0 micrograms/min, or placebo for 125 minutes on different days into six sodium-deprived normal men. During the last 45 minutes of infusion, angiotensin II, 6 ng/kg/min, was infused. Blood pressure, heart rate, plasma concentrations of ANF, aldosterone, and cortisol, and plasma renin activity (PRA) were measured before and during infusion. Steady state mean plasma ANF levels during infusion were 26.2 (placebo), 68.8 (0.1 micrograms ANF/min), 221 (0.3 micrograms ANF/min), and 648 pg/ml (1.0 microgram ANF/min). Systolic blood pressure fell significantly (with 1.0 microgram ANF/min), and diastolic pressure tended to rise in a dose-dependent manner, while heart rate was unchanged. PRA and plasma aldosterone fell during ANF infusion in a dose dependent manner (significant with 0.3 and 1.0 microgram ANF/min infused). The blood pressure-raising and aldosterone-stimulating effects of angiotensin II were blunted by ANF (significant only with 1.0 microgram ANF/min). It is concluded that effects of ANF on blood pressure and the renin-aldosterone system occur with plasma ANF levels close to the physiological range, as well as with slightly elevated ANF levels, as observed in congestive heart failure and renal insufficiency. PMID- 2971620 TI - Iron-regulated hemolysin production and utilization of heme and hemoglobin by Vibrio cholerae. AB - El Tor and non-O1 strains of Vibrio cholerae were analyzed to determine whether synthesis of secreted hemolysin was influenced by the concentration of iron in the medium. Synthesis of hemolysin was found to be iron regulated in both El Tor and non-O1 isolates. Increased levels of hemolytic activity were detected in supernatants of iron-starved cells. Spontaneous hemolysin-deficient mutants of one non-O1 strain were found to occur at high frequency. These variants also failed to synthesize vibriobactin, the iron transport compound utilized by V. cholerae. Another non-O1 strain was found to synthesize both hemolysin and vibriobactin constitutively. When the cloned Escherichia coli fur gene, encoded on the plasmid pABN203, was introduced into this constitutive strain, normal iron regulation of both hemolysin and vibriobactin was reestablished. The ability of V. cholerae to utilize mammalian iron compounds was determined, and it was found that both hemin and hemoglobin could serve as sole sources of iron. PMID- 2971622 TI - Screening models in occupational health practice to detect and to assess individuals and groups at risk because of exposure and/or with decreased capacities to cope with workplace chemicals. PMID- 2971621 TI - Role of immunoglobulin G in platelet aggregation by viridans group streptococci. AB - The aggregation of human platelets by the viridans group streptococci requires both direct platelet-bacterium binding and plasma components. Some of these extracellular constituents (e.g., fibrinogen) are cofactors for ADP, which mediates the terminal events in platelet activation by these organisms. In addition, other plasma components which are specific for viridans group streptococci are necessary. To better define these latter cofactors, we examined the role of immunoglobulin G (IgG) in platelet aggregation by two strains of viridans group streptococci. The addition of either strain to washed human platelets suspended in normal plasma resulted in a 5- to 12-min lag phase, followed by brisk and irreversible platelet aggregation. In contrast, neither strain aggregated platelets suspended in IgG-depleted plasma (IgG concentration, less than or equal to 6.7 micrograms/ml). The addition of IgG (1.0 mg/ml) to the platelet suspension restored normal aggregation. Absorption of the IgG with intact bacteria abolished its ability to support aggregation. Preincubation of washed platelets with a murine monoclonal antibody to the 40,000-Mr platelet Fc receptor blocked aggregation by both strains, but had no effect on aggregation by ADP (5 microM) or collagen (200 micrograms/ml). Neither strain aggregated gel filtered platelets supplemented with fibrinogen (100 micrograms/ml), whereas ADP induced a maximal platelet response. When IgG (1.0 mg/ml) was added to the suspension of gel-filtered platelets, both strains produced normal aggregation. These results indicate that specific IgG is required for platelet aggregation by viridans group streptococci and that platelet activation is mediated through the 40,000-Mr Fc receptor on the platelet surface. PMID- 2971623 TI - Increased immunoglobulin E Fc receptor bearing cells in germinal centers of hyperimmunoglobulinemia E patients. AB - Lymph nodes from 6 patients with hyperimmunoglobulinemia E (hyper-IgE; 4 with Kimura's disease, 1 with atopic dermatitis, and 1 with immunoblastic lymphadenopathy-like T cell lymphoma) and from 7 patients with normal IgE levels were studied to determine the localization of dendritic reticulum cells and of cells bearing Fc epsilon and C3d receptors and immunoglobulin E. The avidin biotin-glucose oxidase method was used for unfixed biopsy specimens. To identify the above-mentioned cells, H107, a murine monoclonal antibody specific to the Fc epsilon receptor molecule, and corresponding antibodies specific to the other cell types were used. In 5 hyper-IgE patients (4 with Kimura's disease and 1 with atopic dermatitis) all germinal centers of the lymph nodes showed heavy reticular staining with H107, the dendritic reticulum cells being most intensely stained. In contrast, the germinal centers of the lymph nodes in the 7 patients with a normal IgE level only lightly or partially stained with H107. The staining pattern of anti-IgE was similar to that of H107 in the hyper-IgE cases. Likewise, C3d receptor and dendritic reticulum cell-related antigens were demonstrated very intensely in all germinal centers in lymph nodes of patients with hyper-IgE and normal IgE levels. These findings suggest that in a hyper-IgE state increased numbers of dendritic reticulum cells in a germinal center express the Fc epsilon receptor and that such cells may play a role in the differentiation of IgE producing memory B cells. PMID- 2971624 TI - Inhibition of eosinophil chemotaxis by a new antiallergic compound (cetirizine). AB - The in vivo inhibitory effect of a new antiallergic, anti-H1 drug, cetirizine, on eosinophil attraction at skin sites challenged with various stimuli has been recently suggested. In the present work, we confirmed that this molecule, at therapeutical concentration, has a potent inhibitory action on eosinophil response to different chemoattractant mediators such as platelet-activating factor (PAF acether) and N-formyl methionyl leucyl phenyl alanyl in vitro. Another anti-H1 drug, polaramine, did not show this effect at the same concentration. These findings suggest that cetirizine in addition to its antihistaminic effect could also play a direct inhibitory effect on eosinophil recruitment. Moreover, cetirizine was not toxic for eosinophils and did not induce degranulation, as shown by the absence of peroxidase release. Comparison between cetirizine and a PAF acether antagonist (BN 52021) suggested that cetirizine did not act by a PAF receptor-blocking activity. PMID- 2971625 TI - Endocrine, seminal and peripheral effects of depot medroxyprogesterone acetate and testosterone enanthate in men. AB - Depot medroxyprogesterone acetate (D-MPA, 250 mg) and testosterone enanthate (TE, 200 mg) were administered twice with a 4-week interval to nine healthy men, and the levels in blood of steroids, gonadotrophins, lipoproteins, sex hormone binding globulin (SHBG) and prostaglandins (PGs) were measured, as well as steroid levels in semen and the sperm count and motility. The hormones analysed were: MPA, testosterone, androstenedione (A), dihydrotestosterone (DHT), oestradiol (E2), cortisol (C), luteinizing hormone (LH), follicle stimulating hormone (FSH) and the sulphoconjugated forms (-S) of testosterone, DHT, pregnenolone (5-P) and dehydroepiandrosterone (DHEA). Peak values of MPA (10.2 +/ 4.6 nmol/l) and testosterone (28.0 +/- 10.0) were found in the first blood samples 2 days after each injection. Thereafter the levels of MPA decreased gradually and reached the limit of detection 18-20 weeks after the second injection. Blood levels of testosterone fell sharply from the peak values and were grossly subnormal 2 weeks after each injection; levels did not return to pretreatment values during 24 weeks of follow-up. The pattern of change of DHT, A, E2 and sulphonated androgens was similar to that of testosterone. These data suggest that D-MPA and TE are absorbed at similar rates, and that the TE is metabolized rapidly. The subsequent reduction in the levels of A, testosterone-S and DHT-S was less marked and reached pretreatment values earlier than did the testosterone levels. No obvious changes were found in the levels of C, 5-P-S and DHEA-S or in the seminal plasma levels of the various steroids studied. The blood levels of LH and FSH fell precipitously 2 days after the first injection, then started to increase 4 weeks after the second injection to reach pretreatment values 12 weeks later. Of the lipoproteins studied only the levels of HDL cholesterol and SHBG were found suppressed after treatment. Severe oligozoospermia and the complete absence of progressively motile sperm, in at least one semen sample, was observed in all subjects at 3-7 and at 5-16 weeks, respectively, after the last injection, suggesting that the men were infertile for at least 1 month after treatment. A spurious increase in the PG content of semen was also observed. In spite of the low blood testosterone levels, no subject reported changes in sexual behaviour or other signs of anabolic imbalance during or after the study. However, the increase in levels of E2 in some individuals should be kept in mind as a possible cause of side-effects.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2971627 TI - Differentiation of the human pancreatic adenocarcinoma cell line (Capan-1) in culture and co-culture with fibroblasts dome formation. AB - This study was designed to investigate differentiation of human pancreatic duct carcinoma cells (Capan-1) in vitro. Observations on live cells, and electron microscopic examination, together with enzymological and immunocytochemical methods, have demonstrated that these cells differentiate spontaneously at an early stage. The cells are seen to be joined by apical junctions. High ATPase activity can be detected in the basolateral membranes, and the cells secrete a gastric type mucin (MI) bearing acidic groups. During differentiation in culture, they form domes which are thought to be the morphological expression of trans epithelial transport of water and electrolytes. This particular structure is transitory, since after 6 days in culture all the cells lose their adhesivity, and form into floating cords. Co-culture of Capan-1 cells and human, nude mice or chick embryo fibroblasts leads to a higher degree of differentiation of epithelial cells, reflected by the earlier appearance of numerous domes. In addition, the anchorage of Capan-1 cells to fibroblasts prevents retraction of the monolayer, and enables the domes to be maintained in the cultures for more than one month. These findings suggest that Capan-1 cells are able to carry out trans-epithelial movement of water and electrolytes. It is suggested that excretion of ions (bicarbonate and/or chloride) is preserved after transformation of pancreatic duct cells. Mucins (MI) and the recently described VIP receptor sites are also thought to play a part in these exchange processes. PMID- 2971626 TI - Establishment and characterization of three transplantable EBV-containing nasopharyngeal carcinomas. AB - Three transplantable nasopharyngeal carcinoma (NPC) tumors, designated C15, C17 and C18, have been obtained and characterized. C15, derived from a primary NPC tumor, has been propagated in nude mice for 30 passages. C17 and C18, derived from metastatic NPC tissue, have been passaged 10 times. Desmosomes, present in every case, provided confirmation of the epithelial origin of all 3 tumors. The Epstein-Barr virus (EBV) genome is contained in C15, C18 and C17 tumor cells with 30, 12 and 3 copies, respectively. The Epstein-Barr virus nuclear antigen (EBNA) was stained by the classical anti-complement immunofluorescence (ACIF) technique. Fluorescence intensity was strong in C15, moderate in C18, and hardly detectable in C17 cells. No expression of the EA and VCA antigens was detected. Flow cytometry analysis performed on monocellular suspensions showed the absence of detectable CR2 molecules (the EBV receptor on B lymphocytes) in all 3 tumors, and the constitutive expression of HLA class-II antigens in C15 and C17 cells. IL-1 activity was demonstrated in the supernatant of C15 and C17 cells cultivated in vitro for 3 days. These data confirm that the constitutive synthesis of MHC class II molecules and the release of IL-1-like activities are frequent features of NPC cells. These characteristics could be of importance in relation with the T-cell infiltrate found in NPC primary tumors. PMID- 2971628 TI - Symmetrical bronchial pattern with normal atrial morphology. AB - An infant with trisomy 21 and tracheostenosis is described. Postmortem findings included symmetrical bronchi, normal atrial arrangement ("situs") and anomalous thoracic vessels. The lack of concordance between the bronchial morphology and atrial arrangement supports the premise that atrial morphology is a more accurate predictor of the presence or absence of the isomerism syndromes. PMID- 2971629 TI - Suppression of yeast ingestion by dexamethasone in macrophage cultures: evidence for a steroid-induced phagocytosis inhibitory protein. AB - The mechanism by which glucocorticoid steroids suppress yeast phagocytosis in cultures of resident and thioglycollate-elicited murine peritoneal macrophages was examined. Time course and dose-response studies demonstrated that the phagocytic capacity of resident macrophages was suppressed by dexamethasone to the same extent in both newly established cultures and cultures that were incubated for several days. In contrast, relative to newly established cultures of elicited cells that were treated with the drug, elicited macrophages that were incubated for at least 1 day prior to exposure to dexamethasone, exhibited enhanced sensitivity to the action of the steroid. Steroid-induced phagocytic inhibitory responses were blocked by the metabolic inhibitors cycloheximide and actinomycin D. The suppression of phagocytosis by dexamethasone was mediated by a factor, present in the cellular homogenates of steroid-treated macrophages, that was partially purified by Sephadex G-25 chromatography. Since the phagocytic inhibitory activity in these homogenates was destroyed following exposure to heat and trypsin, the factor has been named phagocytosis inhibitory protein (PIP). The antiphagocytic activity of PIP was neutralized by treatment with RM23, a monoclonal antibody directed against lipocortin. The results support the hypothesis that the suppression of yeast ingestion is mediated by the action of PIP, which is induced in dexamethasone-treated macrophage cultures. Moreover, PIP appears to belong to the lipocortin family of phospholipase inhibitory proteins. PMID- 2971630 TI - Immunological studies on paroxetine, a novel anti-depressant drug. AB - Paroxetine is a novel and selective neuronal 5-hydroxy-tryptamine uptake inhibitor with anti-depressant activity. Paroxetine was examined for its ability to induce adverse immunological reactions, either as a consequence of a specific immune response or by a direct or indirect effect on the immune system. Paroxetine did not react in vitro with protein amino or thiol groups, suggesting that it lacks the capacity to form potentially immunogenic hapten protein conjugates. No anti-paroxetine antibody was detected in plasma or serum samples from patients and rats following oral administration over prolonged periods, or from epicutaneously exposed guinea pigs, or from rabbits given paroxetine in Freund's adjuvant, suggesting that paroxetine does not have the capacity to elicit humoral immune responses. Guinea pigs epicutaneously exposed to paroxetine did not develop contact sensitivity, suggesting that it does not have the capacity to elicit cell-mediated immune responses. These results suggest that paroxetine lacks intrinsic immunogenicity. Anti-SRBC antibody plaque-forming cell responses in mice were unaffected by oral administration of paroxetine, and paroxetine had no significant effect on ex vivo and in vitro murine macrophage phagocytosis of opsonized SRBC or on ex vivo murine splenocyte mitogen responses, suggesting that paroxetine does not exert modulatory effects on the immune system or on macrophage function. These findings, together with the results of pre clinical safety evaluation studies, suggest that paroxetine is unlikely to have immunotoxic effects. PMID- 2971631 TI - Immunomodulating properties of cianidanol on responsiveness and function of human peripheral blood T-cells and K-cells. AB - Cianidanol (Ci) [(+)-catechin] is a lipophilic compound which interacts with membrane lipids and affects responsiveness and function of immunocompetent cells. We therefore studied the immunomodulating properties of Ci on the proliferative response of human peripheral T-cells in one-way mixed lymphocyte reaction (1-MLR) and autologous mixed lymphocyte reaction (AMLR); on the generation of cytotoxic T cells (Tc-cells), suppressor T-cells (Ts-cells, comprising radiosensitive as well as radioresistant suppressor T-cells) and radioresistant suppressor T-cells (rrTs) in 1-MLC; and on the cytolytic activity of Tc-cells and K-cells. In 1-MLR we observed a small stimulation of cell proliferation at Ci concentrations up to 108 microM whereas higher concentrations led to a marked suppression (100% at 435 microM). The generation of Ts-cells and rrTs-cells in 1-MLR was clearly suppressed at Ci-concentrations above 435 microM and 108 microM, respectively. The timing of Ts-cell formation was not influenced. The Tc-cell generation in 1 MLR was inhibited at high doses, and at 870 microM 59% suppression was observed. A similar dose-dependent suppressive effect of Ci was seen by testing for the cytolytic activity of ADCC-reactive K-cells and of CML-reactive Tc-cells generated in 1-MLR in the absence of Ci. At the highest concentration used (870 microM) the CML was suppressed by 45% and the ADCC by 46%. Our investigation on Ci's influence on the efferent and afferent part of immune responses in vitro demonstrated both stimulatory and inhibitory effects usually occurring at low and high concentrations, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971632 TI - Polyostotic fibrous dysplasia with contrasting responses to calcitonin and mithramycin: aetiological and therapeutic implications. PMID- 2971633 TI - Suppression of allograft immunity by 3,4,5,3',4',5'-hexachlorobiphenyl. II. Effects of exposure on mixed lymphocyte reactivity and induction of suppressor cell activity in vitro. AB - Previous studies in our laboratory have established the sensitivity of the in vivo allogeneic cytotoxic T lymphocyte (CTL) response to suppression by 3,4,5,3',4',5'-hexachlorobiphenyl[(345)2-HxCB], a toxic, Ah receptor-binding polychlorinated biphenyl isomer. The present studies have examined possible cellular mechanisms for this suppression. A modest dose-dependent suppression of the proliferative response to alloantigen in mixed lymphocyte culture (MLC) was observed with lymphocytes from B6 mice exposed to 10 or 100 mg/kg (345)2-HxCB while the CTL response generated in MLC was significantly suppressed only following exposure to 100 mg/kg (345)2-HxCB. The amount of time between treatment with (345)2-HxCB and sacrifice, which ranged from 2 to 23 days, did not appear to influence the degree of immunosuppression produced by (345)2-HxCB exposure. Mitomycin C-treated lymphocytes from B6 mice treated with (345)2-HxCB were not suppressive when added as third party cells to an independent MLC. However, if the mice were alloimmune, lymphocyte-mediated suppression of the MLC response was observed and directly correlated with the magnitude of the CTL response present in the same population. Thus, (345)2-HxCB-treated mice which had less CTL activity as compared to vehicle-treated mice also had less suppressor activity. Further analysis indicated that stimulator cell lysis by the CTL was likely to be responsible for the inhibitory activity of the alloimmune lymphocytes rather than suppressor cells per se. Avoidance of stimulator cell lysis by using H-2 incompatible MLC stimulator cells revealed the existence of antigen-nonspecific suppressor activity that was greater with lymphocytes from vehicle-treated than from (345)2-HxCB-treated mice, suggesting that both CTL and suppressor cell activities were suppressed by (345)2-HxCB exposure. Direct addition of (345)2 HxCB to lymphocyte cultures in vitro indicated a lack of direct toxicity of (345)2-HxCB on lymphoproliferative responses to mitogen or alloantigen at concentrations equal to or less than 1 x 10(-6) M. Thus, the in-vitro functional integrity of lymphocytes obtained from (345)2-HxCB-treated mice coupled with the lack of a direct lymphotoxic effect of (345)2-HxCB in vitro suggest an indirect mechanism of action for (345)2-HxCB-mediated suppression of CTL activity in vivo. Previous reports implicating suppressor cell induction and/or activation by Ah receptor-binding halogenated aromatic hydrocarbons that mediate the inhibition of CTL generation were not confirmed in these studies. PMID- 2971634 TI - Tetranactin, a macrotetrolide antibiotic, suppresses in vitro proliferation of human lymphocytes and generation of cytotoxicity. AB - Tetranactin, a hydrophobic cyclic antibiotic produced by Streptomyces aureus, has previously been shown to suppress in vitro activation of rat lymphocytes by concanavalin A as well as the onset of experimental autoimmune uveoretinitis in Lewis rats. Here we report the effects of tetranactin on human T and NK lymphocytes in vitro. Tetranactin, at concentrations up to 100 ng/ml, was not toxic to human lymphocytes but completely abrogated the proliferation of human T lymphocytes in response to allogeneic cells in mixed lymphocyte cultures. Tetranactin also blocked the initiation of proliferation in response to interleukin-2, but did not block proliferation of interleukin-2-activated cells. Tetranactin also blocked generation of cytotoxic T lymphocytes and activated killer cells in the mixed lymphocyte culture. However, up to 100 ng/ml tetranactin did not alter the lytic activity of cytotoxic T or NK lymphocytes generated in its absence. The ability of low doses of tetranactin to block the induction of lymphoproliferation is similar to the action of cyclosporin A. Since cyclosporin A is also a cyclic hydrophobic molecule, the immunosuppressive actions of these two agents may involve a similar mechanism. PMID- 2971636 TI - Immunohistochemical localization of atrial natriuretic polypeptide (ANP) in human atrial and ventricular myocardiocytes. AB - To date, there have been few immunohistochemical investigations of atrial natriuretic polypeptide (ANP) in human cardiac tissue, especially the ventricles. In this study, myocardial tissue was obtained from two sources: the bilateral atria and ventricles at autopsy; and biopsy tissues from the right auricle and left ventricle of a patient with myocardial infarction undergoing surgery. These tissues were examined by the avidin-biotin immunoperoxidase technique using three kinds of primary ANP-antibodies. ANP-immunoreactivity was observed in the perinuclear region of myocytes of all tissues examined. The intensity of the reaction was stronger in atrial tissue, weaker in ventricular tissue. In the later tissue, the positive-staining myocytes were not part of the pulse conducting system. Although the tissues we studied were not obtained from normal hearts, our data demonstrates that ANP-reactivity can be detected in ventricular myocytes outside the pulse-conducting system. PMID- 2971635 TI - Immunomodulatory activity and non-specific suppressor cell generation by spirogermanium in murine and rat models of cell-mediated immunity. AB - Spirogermanium (SG) is a metal-containing compound reported to have antitumor, antiarthritic, antimalarial and immunoregulatory activity. In this study we have demonstrated that treatment of mice and rats with spirogermanium results in an inhibition of autoimmune disease and cell-mediated immune (CMI) responses. Prophylactic administration of SG inhibited the development of adjuvant-induced arthritis and the DTH response to purified protein derivative (PPD) in Lewis strain rats. SG treatment was also able to alleviate the symptoms of experimental autoimmune encephalomyelitis (EAE) induced in Lewis rats. In two strains of mice, BDF1 and C57B1/6, the DTH response to sheep red blood cells could be suppressed by intraperitoneal (i.p.) administration of SG. The spleens of both mice and rats that have been treated with this drug contain suppressor cells which inhibit the response of normal cells to concanavalin A (Con A) and the mixed lymphocyte reaction. In addition, the generation of cytotoxic T cells (CTL) in the murine MLR is abrogated in the presence of these suppressor cells. The suppressor cells were radiation-resistant (2000 rad), indomethacin-insensitive and were not depleted by treatment with anti-Thy-1.2 antiserum plus complement. These results suggest that SG modulates cell-mediated immune responses in vivo by the induction of non-specific suppressor cells. PMID- 2971637 TI - Diagnostic laparoscopy in apparent uterine agenesis. AB - We report a case of an adolescent female with apparent agenesis of the uterus with chromosomally competent ovarian dysgenesis. In our case, the apparent absence of the uterus on the laparoscopy constituted a severe hypoplasia of the organ prior to estrogen stimulation, rather than aplasia. Following a year of estrogen/progesterone replacement, an infantile but otherwise normal uterus could be demonstrated by hysterosalpinogogram. In gonadal dysgenesis, therefore, it is not possible to differentiate the absence of the uterus from severe hypoplasia using laparoscopy. PMID- 2971638 TI - Expression of CR2 (C3d receptor) on the cell membranes of adult T cell leukemia. AB - MT-2 cells, which produce human T-cell lymphotropic virus type I (HTLV-I), are known to have a complement receptor. We have established that the complement receptor is CR2 which binds C3d on immune complexes but not CR1. CR2 was also detected on ATL-3I cells but no complement receptor was detected on ATL-1K cells which lack ATL antigen (ATLA). Since CR2 is not detectable on normal T lymphocytes, the presence of CR2 on some ATL cells might suggest that ATL cells were derived from a particular minor lineage of T cells, or HTLV-I has a capacity to induce CR2, which has been demonstrated to be an a-type growth factor for B lymphocytes and to be a receptor for Epstein-Barr virus. PMID- 2971640 TI - Isolation of 1,5-dideoxy-1,5-imino-D-mannitol from culture broth of Streptomyces species. PMID- 2971639 TI - Increased intratumor concentration of fluorescein-isothiocyanate-labeled neocarzinostatin in rats under angiotensin-induced hypertension. AB - On the basis of the observation that the tumor tissue blood flow selectively increases under angiotensin (AT)-induced hypertension, the change of the drug concentration in the tumor and normal tissues was examined in male Donryu rats. The intratumor concentration of fluorescein isothiocyanate-labeled neocarzinostatin was about 2-fold higher in the AT-induced hypertension group than in the control up to 20 min after the drug injection. In the normal organs or the uninvolved organs of the tumor-bearing rats, however, no clear increase was seen in the experimental group compared with the control, as anticipated from the observation of the tissue blood flow. The present study supports the hypothesis that the enhanced anticancer effect in chemotherapy under AT-induced hypertension formerly reported is due to the tumor-selective enhancement of the drug delivery. PMID- 2971641 TI - Calcium transport mechanism in the endolymph of the chinchilla. AB - The Ca2+ transport mechanism between endolymph and perilymph was evaluated by the effects of vanadate and amiloride on the endocochlear potential (EP) and the Ca2+ concentration in endolymph using Ca2+-selective microelectrodes. Under normal conditions, the EP was 81.8 +/- 0.9 mV, and the Ca2+ concentrations in endolymph and perilymph were 16.6 +/- 1.3 microM and 1.85 +/- 0.11 mM (N = 12), respectively. Therefore, the uphill electrochemical potential gradient for Ca2+ from perilymph to endolymph, 20.2 +/- 2.0 mV, indicates the existence of an active uptake of Ca2+ into endolymph. Vanadate, the inhibitor of Ca2+-ATPase, topically applied to the round window membrane caused biphasic changes of the EP and the endolymph Ca2+ concentration; the former in a transient increase followed by a consistent decrease and the latter in a slow decrease followed by a slow increase. Amiloride induced a slight EP depression and a concomitantly slight elevation of the Ca2+ concentration in endolymph. The electrochemical potential gradient for Ca2+ between endolymph and perilymph vanished with the use of vanadate but was not affected by amiloride. These results suggest that Ca2+ ATPase, sensitive to vanadate, maintained the bulk of active Ca2+ transport in the cochlea and that the participation of Na+-Ca2+ exchange is negligible. PMID- 2971642 TI - Relative strengths of the chest wall muscles. AB - We hypothesized that during maximal respiratory efforts involving the simultaneous activation of two or more chest wall muscles (or muscle groups), differences in muscle strength require that the activity of the stronger muscle be submaximal to prevent changes in thoracoabdominal configuration. Furthermore we predicted that maximal respiratory pressures are limited by the strength of the weaker muscle involved. To test these hypotheses, we measured the pleural pressure, abdominal pressure (Pab), and transdiaphragmatic pressure (Pdi) generated during maximal inspiratory, open-glottis and closed-glottis expulsive, and combined inspiratory and expulsive maneuvers in four adults. We then determined the activation of the diaphragm and abdominal muscles during selected maximal respiratory maneuvers, using electromyography and phrenic nerve stimulation. In all subjects, the Pdi generated during maximal inspiratory efforts was significantly lower than the Pdi generated during open-glottis expulsive or combined efforts, suggesting that rib cage, not diaphragm, strength limits maximal inspiratory pressure. Similarly, at high lung volumes, the Pab generated during closed-glottis expulsive efforts was significantly greater than that generated during open-glottis efforts, suggesting that the latter pressure is limited by diaphragm, not abdominal muscle, strength. As predicted, diaphragm activation was submaximal during maximal inspiratory efforts, and abdominal muscle activation was submaximal during open-glottis expulsive efforts at midlung volume. Additionally, assisting the inspiratory muscles of the rib cage with negative body-surface pressure significantly increased maximal inspiratory pressure, whereas loading the rib cage muscles with rib cage compression decreased maximal inspiratory pressure. We conclude that activation of the chest wall muscles during static respiratory efforts is determined by the relative strengths and mechanical advantage of the muscles involved. PMID- 2971644 TI - Doctors and dependency. PMID- 2971645 TI - Thrombotic thrombocytopenic purpura: the importance of aggressive treatment. PMID- 2971643 TI - Presence of insulinlike growth factor receptors and lack of insulin receptors on fetal bovine smooth muscle cells. AB - Previous investigations have demonstrated specific receptors and associated mitogenic actions for insulin and insulinlike growth factors I and II (IGF-I and II) in postnatal bovine aortic smooth muscle. Using fetal tissue we have observed different patterns of binding and action for these peptides. Smooth muscle cells isolated from near-term fetal bovine aortae were studied in early passage. Specific receptors for both IGF-I and IGF-II were identified. Specific binding averaged 5.7%/2.5 X 10(5) cells for IGF-I, and 16.2% for IGF-II, and 0.3% for insulin. High affinity Kd for both IGF receptors were nanomolar. IGF-II was fivefold less potent than IGF-I in displacing IGF-I binding. IGF-I showed no affinity for the IGF-II receptor. Insulin, at physiologic concentrations, was incapable of displacing either IGF-I or IGF-II binding. Cellular incorporation of [methyl-3H]thymidine was stimulated at the lowest dose of IGF-I tested, 0.5 ng/ml. IGF-II showed no effect up to 100 ng/ml, after which a sharp increase in incorporation was noted. Insulin had a similar effect only at concentrations greater than 0.5 micrograms/ml, with a maximal response noted at 5 to 10 micrograms/ml. Our results indicate that fetal bovine aortic smooth muscle cells have an abundance of IGF receptors but lack specific insulin receptors. In addition, IGF-II binding levels are three times higher than for IGF-I. These results are consistent with observations in other species, in which a predominance of IGF over insulin receptors has been demonstrated in fetal tissue, and provide further evidence for a role for the IGFs in embryonic cellular metabolism. PMID- 2971646 TI - Treatment of recurrent Hodgkin's disease. PMID- 2971648 TI - Identification of troponin-I of crayfish myofibrils. AB - Crayfish (Procambarus clarkii) myofibrils contain two basic proteins of molecular weights of 25,000 and 23,000. Both of the two proteins inhibit actomyosin ATPase as the vertebrate troponin-I does. These results differ from the previous one that troponin-I of crayfish (Astacus leptodactylus) showed a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS PAGE). PMID- 2971647 TI - Siderophore-mediated uptake of Fe3+ by the plant growth-stimulating Pseudomonas putida strain WCS358 and by other rhizosphere microorganisms. AB - Under iron-limited conditions, Pseudomonas putida WCS358 produces a siderophore, pseudobactin 358, which is essential for the plant growth-stimulating ability of this strain. Cells of strain WCS358, provided that they have been grown under Fe3+ limitation, take up 55Fe3+ from the 55Fe3+-labeled pseudobactin 358 complex with Km and Vmax values of 0.23 microM and 0.14 nmol/mg of cell dry weight per min, respectively. Uptake experiments with cells treated with various metabolic inhibitors showed that this Fe3+ uptake process was dependent on the proton motive force. Furthermore, strain WCS358 was shown to be able to take up Fe3+ complexed to the siderophore of another plant-beneficial P. fluorescens strain, WCS374. The tested pathogenic rhizobacteria and rhizofungi were neither able to grow on Fe3+-deficient medium in the presence of pseudobactin 358 nor able to take up 55Fe3+ from 55Fe3+-pseudobactin 358. The same applies for three cyanide producing Pseudomonas strains which are supposed to be representatives of the minor pathogens. These results indicate that the extraordinary ability of strain WCS358 to compete efficiently for Fe3+ is based on the fact that the pathogenic and deleterious rhizosphere microorganisms, in contrast to strain WCS358 itself, are not able to take up Fe3+ from Fe3+-pseudobactin 358 complexes. PMID- 2971649 TI - Preparation of subfragment-1 from abalone smooth muscle myosin. AB - Myosin subfragment-1 (S1), which has one heavy chain (HC) (93 kDa) and two light chains (LC1 and LC2), was prepared by papain digestion of myosin from abalone smooth muscle in the presence of Ca2+. The Ca-sensitivity of abalone S1 itself was not lost completely (about 30%). The tryptic digestion of S1 showed that in the presence of EDTA, S1 HC was split into 68, 55, and 23 kDa fragments, as in the presence of Ca2+, but 23 kDa was further degraded into 19 kDa. In contrast to the result in the presence of Ca2+, LCs disappeared in the early stage of reaction and Ca-ATPase activity decreased rapidly to about 70% of that of intact S1. This rapid decrease of Ca-ATPase activity seemed to be accompanied with the digestion of LCs. Therefore, LCs contribute to the protection of 23 kDa fragment from further digestion, to the maintenance of Ca-ATPase activity by stabilizing the structure of S1 to some extent in the presence of Ca2+. Since F-actin suppressed the cleavage of S1 HC to 68 and 23 kDa during tryptic digestion, it might be that 23 and 68 kDa corresponded to 20 kDa (C-terminal fragment) and to 50 + 25 kDa (N-terminal fragment) of skeletal myosin S1, respectively. PMID- 2971650 TI - Deficient polymerization in vitro of a point-mutated beta-actin expressed in a transformed human fibroblast cell line. AB - HUT-14 cells, tumorigenic human fibroblasts, express a mutant beta-actin which has a single amino acid substitution at position 244 (glycine to aspartic acid), in addition to normal beta- and gamma-actin. In order to characterize the biochemical function of the mutant beta-actin, actins were extracted and purified from HUT-14 cells. The partially purified actin fraction contained beta-, gamma-, and mutant beta-actins in the ratio of 1:1:1, the same ratio as in the cells. When the actin of this fraction was purified through a polymerization step, mutant beta-actin was always less incorporated into actin filaments than beta- and gamma-actin. When the polymerization ability of purified HUT-14 actins was examined by sedimentation technique, it was lower than those of muscle and of cytoplasmic actins from another human cell line (HUT-11) which expresses only normal beta- and gamma-actin, in the ratio of 2:1. The deficient polymerization of mutant beta-actin was also observed by examining the ratio of beta-, gamma-, and mutant beta-actins incorporated into actin filaments. The ratio of mutant beta-actin in polymerized actins under all conditions examined was always less than that before polymerization. These results indicate that the single amino acid substitution at position 244 caused the reduction of incorporation of the mutant beta-actin into actin filaments in vitro. PMID- 2971651 TI - Isolation of genes encoding the Neurospora vacuolar ATPase. Analysis of vma-1 encoding the 67-kDa subunit reveals homology to other ATPases. AB - The vacuolar membrane of Neurospora crassa contains a H+-translocating ATPase composed of at least three subunits with approximate molecular weights of 70,000, 60,000, and 15,000. Both genomic and cDNA clones encoding the largest subunit, which appears to contain the active site of the enzyme, have been isolated and sequenced. The gene for this subunit, designated vma-1, contains six small introns (60-131 base pairs) and encodes a hydrophilic protein of 607 amino acids, Mr 67,121. Within the sequence is a putative nucleotide-binding region, consistent with the proposal that this subunit contains the site of ATP hydrolysis. This 67-kDa polypeptide shows high homology (62% identical residues overall and 84% in the middle of the protein) to the analogous polypeptide of a higher plant vacuolar ATPase. The hypothesis that the vacuolar ATPase is related to F0F1 ATPases is strongly supported by the finding of considerable homology between the 67-kDa subunit of the Neurospora vacuolar ATPase and both the alpha and beta subunits of F0F1 ATPases. PMID- 2971652 TI - The actin-myosin subfragment-1 complex stabilized by phenyldiglyoxal. AB - Nuclear magnetic resonance studies have revealed the importance of arginine residues in the actin-myosin interface (Moir, A. J. G., and Levine, B. A. (1986) J. Inorg. Chem. 27, 271-278). In the present study, we tested the involvement of these residues in the rigor complex between actin and subfragment-1 (S1) by chemical cross-linking experiments using phenyldiglyoxal. Two kinds of linkages were observed, one within the S1 heavy chain itself (120-kDa product) and the other between actin and the S1 heavy chain (200-kDa product). The phenyldiglyoxal had an effect similar to that of phenylglyoxal on S1 ATPase activities. We also show that phenyldiglyoxal (of 0.6-0.8 nm arm length) cross-links an arginine residue of the 50-kDa domain to one in the 20-kDa domain of the S1 heavy chain in the absence of actin or to an arginine in actin when actin is present. The presence of Mg2+, adenosine 5'-diphosphate or 5'-adenylyl imidodiphosphate suppressed the intermolecular linkage with actin, and favored the intramolecular cross-link, (i.e. between 50-kDa and 20-kDa fragments). We propose that the same arginyl residue in the N-terminal part of the 50-kDa domain can be cross-linked to a nearby arginine in either the 20-kDa domain or the actin molecule. In accordance with the amino acid sequence of each protein this also implies that the actin-myosin interaction involves arginine residues located either after residue 28 of the N-terminal part of actin, since this actin region is devoid of arginine residues, or in the N-terminal portion of the 50-kDa domain, i.e. between residues 239 and 455. PMID- 2971653 TI - Purification of a 31,000-dalton insulin-like growth factor binding protein from human amniotic fluid. Isolation of two forms with different biologic actions. AB - Human amniotic fluid has been shown to contain a protein that binds insulin-like growth factor I and II (IGF-I and IGF-II). Partially purified preparations of this protein have been reported to inhibit the biologic actions of the IGFs. In these studies our laboratory has used a modified purification procedure to obtain a homogeneous preparation of this protein as determined by polyacrylamide gel electrophoresis and amino acid sequence analysis. During purification the ion exchange chromatography step resulted in two peaks of material with IGF binding activity termed peaks B and C. Each peak was purified separately to homogeneity. Both peaks were estimated to be 31,000 daltons by polyacrylamide gel electrophoresis and their amino acid compositions were nearly identical. Amino acid sequence analysis showed that both peaks had identical N-terminal sequences through the first 28 residues. Neither protein had detectable carbohydrate side chains and each had a similar affinity for radiolabeled IGF-I (1.7-2.2 x 10(10) liters/mol). In contrast, these two forms had marked differences in bioactivity. Concentrations of peak C material between 2 and 20 ng/ml inhibited IGF-I stimulation of [3H]thymidine incorporation into smooth muscle cell DNA. In contrast, when peak B (100 ng/ml) was incubated with IGF-I there was a 4.4-fold enhancement of stimulation of DNA synthesis. Additionally, pure peak B was shown to adhere to cell surfaces, whereas peak C was not adherent. The non-adherent peak C inhibited IGF-I binding to its receptor and to adherent peak B. We conclude that human amniotic fluid contains two forms of IGF binding protein that have very similar physiochemical characteristics but markedly different biologic actions. Since both have similar if not identical amino acid compositions, N terminal sequences, and do not contain carbohydrate, we conclude that they differ in some other as yet undefined post-translational modification. PMID- 2971655 TI - Translocation of a fragment of invertase across microsomal vesicles isolated from Neurospora crassa requires the hydrolysis of a nucleoside triphosphate. AB - The step which requires the hydrolysis of a nucleoside triphosphate for translocation of a protein across microsome was investigated by studying translocation uncoupled from translation using two truncated products of invertase: one product contains the first 262 amino acids of the secreted invertase (Inv262); the other, the first 104 amino acids (Inv104). The truncated products were translated from RNA transcripts without a stop codon. It is demonstrated that the translated products contain an associated ribosome, and the associated ribosome is essential for the posttranslational translocation phenomenon. Also, it is demonstrated that binding of Inv262 to microsomal vesicles made from the cell wall-less mutant of Neurospora crassa does not require the hydrolysis, or the presence, of a nucleoside triphosphate. However, the posttranslational translocation across the membrane, as monitored by glycosylation of the translocated polypeptide chain, does. The data suggest that a nucleoside triphosphatase activity is associated with the translocation of invertase across the membrane. Furthermore, it is demonstrated that binding, and subsequent translocation across the membrane, is dependent on trypsin-sensitive membrane component(s). PMID- 2971654 TI - Identification of mannose 6-phosphate in two asparagine-linked sugar chains of recombinant transforming growth factor-beta 1 precursor. AB - Recombinant transforming growth factor-beta 1 (TGF-beta 1) precursor produced and secreted by a clone of Chinese hamster ovary cells was found to be glycosylated and phosphorylated. Treatment of 32P-labeled precursor protein with N-glycanase indicated that phosphate was incorporated into asparagine-linked complex carbohydrate moieties. Fractionation of 32P-labeled glycopeptides followed by amino acid sequence analysis indicated that greater than 95% of the label was incorporated into two out of three glycosylation sites at Asn-82 and Asn-136 of the TGF-beta 1 precursor. Two-dimensional electrophoretic analysis of acid hydrolyzed precursor protein and precursor protein-derived glycopeptides indicated that 32P was incorporated as mannose 6-phosphate. Binding studies with the purified receptor for mannose 6-phosphate indicated that the TGF-beta 1 precursor could bind to this receptor and the binding was specifically inhibited with mannose 6-phosphate. PMID- 2971656 TI - Mechanism of the cytotoxic synergism of fluoropyrimidines and folinic acid in mouse leukemic cells. AB - We have investigated the mechanism by which reduced folates, such as folinic acid, enhance the cytotoxicity of fluoropyrimidines in L1210 mouse leukemic cells. Exposure of L1210 cells to folinic acid resulted in expansion of intracellular pools of 5,10-CH2-H4PteGlun, delayed the reappearance of catalytically active thymidylate synthase (TS) following 5-fluoro-2'-deoxyuridine exposure, and stabilized inhibited TS complexes over the same concentration range that augmented the cytotoxic effects of fluorodeoxyuridine and 5-fluorouracil. The data showed that, in intact L1210 cells, fluorodeoxyridylate behaves as an inhibitor whose complexes with TS dissociated with a biologically significant rate. However, these complexes become functionally irreversible in cells incubated with high levels of folinic acid. It was also found that bound and total TS levels increased in cells treated with fluorodeoxyuridine to an extent that substantially exceeded the increase in protein content per cell under the same conditions. These results are in accord with the concept that folinic acid augments the effects of the fluoropyrimidines by expansion of cellular 5,10-CH2 H4PteGlun pools with subsequent stabilization of ternary complexes among 5-fluoro 2'-deoxyuridine 5'-monophosphate, TS, and 5,10-CH2-H4PteGlun. In light of the accumulation of TS that occurs following exposure to fluoropyrimidines, this stabilization may be needed for efficient tumor cell killing by these agents. PMID- 2971657 TI - Ankyrin-independent membrane protein-binding sites for brain and erythrocyte spectrin. AB - Brain spectrin reassociates in in vitro binding assays with protein(s) in highly extracted brain membranes quantitatively depleted of ankyrin and spectrin. These newly described membrane sites for spectrin are biologically significant and involve a protein since (a) binding occurs optimally at physiological pH (6.7 6.9) and salt concentrations (50 mM), (b) binding is abolished by digestion of membranes with alpha-chymotrypsin, (c) Scatchard analysis is consistent with a binding capacity of at least 50 pmol/mg total membrane protein, and highest affinity of 3 nM. The major ankyrin-independent binding activity of brain spectrin is localized to the beta subunit of spectrin. Brain membranes also contain high affinity binding sites for erythrocyte spectrin, but a 3-4 fold lower capacity than for brain spectrin. Some spectrin-binding sites associate preferentially with brain spectrin, some with erythrocyte spectrin, and some associate with both types of spectrin. Erythrocyte spectrin contains distinct binding domains for ankyrin and brain membrane protein sites, since the Mr = 72,000 spectrin-binding fragment of ankyrin does not compete for binding of spectrin to brain membranes. Spectrin binds to a small number of ankyrin independent sites in erythrocyte membranes present in about 10,000-15,000 copies/cell or 10% of the number of sites for ankyrin. Brain spectrin binds to these sites better than erythrocyte spectrin suggesting that erythrocytes have residual binding sites for nonerythroid spectrin. Ankyrin-independent-binding proteins that selectively bind to certain isoforms of spectrin provide a potentially important flexibility in cellular localization and time of synthesis of proteins involved in spectrin-membrane interactions. This flexibility has implications for assembly of the membrane skeleton and targeting of spectrin isoforms to specialized regions of cells. PMID- 2971659 TI - Identification of the C3b receptor-binding domain in third component of complement. AB - We report here that complement receptor type one (CR1) binds to a region of C3b that is contained within the NH2 terminus of the alpha' chain. In an enzyme linked immunosorbent assay, CR1 bound to C3b, iC3b, and C3c but not to C3d, and this binding was inhibited by soluble C3b and C3c. Further attempts to generate a small C3 fragment capable of binding CR1 were unsuccessful. However, elastase degradation of C3 generated four species of C3c (C3c I-IV), two of which bound CR1. NH2-terminal sequence analysis and sodium dodecyl sulfate-gel electrophoresis of the C3cs indicated that the beta chains and the 40,000-dalton COOH-terminal alpha' chain fragments were identical; the NH2-terminal alpha' chain fragments of C3c I-IV varied from 21,000 to 27,000 daltons and accounted for the differential binding to CR1. C3c-I and II, which do not bind CR1, were missing 8 and 9 residues from the NH2 terminus of the alpha' chain when compared with the intact alpha' chain of C3b. C3c-III and IV, which bind CR1, had NH2 termini identical to the intact NH2-terminal alpha' chain of C3b. Using iodinated concanavalin A and endoglycosidase H, we showed that the NH2-terminal alpha' chains of C3c-I and III were glycosylated, while C3c-II and IV were not. Therefore, these data indicated that the amino terminus of the NH2-terminal alpha' chain fragment of C3c was responsible for binding CR1 while the COOH terminus of this fragment was not involved since the presence or absence of this region in C3c did not affect CR1 binding to C3c. Subsequently, two peptides were synthesized from the NH2-terminal alpha' chain fragment of C3c: X42, 42 residues in length from the NH2 terminus and C30, 30 residues in length from the COOH terminus. X42 inhibited binding of CR1 to C3b, and this effect was also observed with antipeptide antibodies against the X42 peptide. The C30 and other C3-derived peptides and antipeptide antibodies had no effect on the binding of CR1 to C3b. PMID- 2971658 TI - Cyclosporin A-binding protein (cyclophilin) of Neurospora crassa. One gene codes for both the cytosolic and mitochondrial forms. AB - Cyclophilin (cyclosporin A-binding protein) has a dual localization in the mitochondria and in the cytosol of Neurospora crassa. The two forms are encoded by a single gene which is transcribed into mRNAs having different lengths and 5' termini (approximately 1 and 0.8 kilobases). The shorter mRNA specifies the cytosolic protein consisting of 179 amino acids. The longer mRNA is translated into a precursor polypeptide with an amino-terminal extension of 44 amino acids which is cleaved in two steps upon entry into the mitochondrial matrix. Neurospora cyclophilin shows about 60% sequence homology to human and bovine cyclophilins. PMID- 2971660 TI - Kinetics of inactivation of membrane-bound factor Va by activated protein C. Protein S modulates factor Xa protection. AB - Kinetic analyses were done to determine what effect factor Xa and protein S had on the activated protein C (APC)-catalyzed inactivation of factor Va bound to phospholipid vesicles or human platelets. In the presence of optimal concentrations of phospholipid vesicles and Ca2+, a Km of 19.7 +/- 0.6 nM factor Va and a kcat of 23.7 +/- 10 mol of factor Va inactivated/mol of APC/min were obtained. Added purified plasma protein S increased the maximal rate of factor Va inactivation only 2-fold without effect on the Km. Protein S effect was unaltered when the phospholipid concentration was varied by 2 orders of magnitude. The reaction on unactivated human platelets yielded a Km = 12.5 +/- 2.6 nM and kcat = 6.2 +/- 0.6 mol of factor Va inactivated/mol of APC/min. Added purified plasma protein S or release of platelet protein S by platelet activation doubled the kcat value without affecting the Km. Addition of a neutralizing anti-protein S antibody abrogated the effect of plasma protein S or platelet-released protein S, but was without effect in the absence of plasma protein S or platelet activation. Studies with factor Xa indicated that factor Xa protects factor Va from APC catalyzed inactivation by lowering the effective concentration of factor Va available to interact with APC. From these data a dissociation constant of less than 0.5 nM was calculated for the interaction of factor Xa with membrane-bound factor Va. Protein S abrogated the ability of factor Xa to protect factor Va from inactivation by APC without affecting the interaction of factor Xa with factor Va. These combined data suggest that one physiological function of protein S is to allow the APC-catalyzed inactivation of factor Va in the presence of factor Xa. PMID- 2971661 TI - Isolation and partial characterization of high affinity laminin receptors in neural cells. AB - Neural cells in culture (NG-108, PC12, chick dorsal root ganglion, chick spinal cord, and rat astrocytes) bind laminin with an apparent Kd of congruent to 10(-9) M. Laminin affinity chromatography of chick brain membranes washed with 150 mM NaCl and eluted with 0.2 M glycine buffer, pH 3.5, yields a single protein with an apparent molecular mass of 67 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Isoelectric focusing and peptide mapping indicate that the 67-kDa protein is distinct from bovine serum albumin (68 kDa) but indistinguishable from high affinity laminin receptors isolated from skeletal muscle. After electroblotting onto nitrocellulose paper and probing with 125I-laminin, this putative laminin receptor binds laminin specifically (100 ng/ml). A second protein (congruent to 120-140 kDa) is also detected with 125I laminin (100 ng/ml) in the laminin affinity-purified membrane proteins. Both 67- and congruent to 120-140-kDa proteins can be laminin affinity-purified from cultures enriched for neurons (greater than 90%) following metabolic labeling with [35S]methionine. Our data suggest that neural cells (dorsal root ganglion, central nervous system neurons, astrocytes, and several neural cell lines) have high affinity binding sites for laminin and that two membrane proteins, 67- and congruent to 120-140-kDa, are responsible at least in part for this binding. PMID- 2971662 TI - Studies on the mechanism of Escherichia coli DNA polymerase I large fragment. Effect of template sequence and substrate variation on termination of synthesis. AB - Termination of Escherichia coli DNA polymerase I large fragment after processive synthesis on natural and other well-defined template.primer systems has been examined. We found that after any given deoxynucleoside monophosphate incorporation termination occurs in a nonrandom manner with phi X174 DNA as template: Termination is much more likely at some nucleotide residues along the template than at others. Analysis of these stronger termination sites indicates that the template base:incoming nucleotide combination influences termination. Introduction of a double-stranded region along the phi X174 template induces termination, and reducing dNTP concentrations or substituting 2'-deoxynucleoside 5'-O-(1-thio)triphosphate substrates also increases termination. Observations with the phi X174 DNA template system were extended with a defined template containing 1 inosine residue in an otherwise d(T)n homopolymer. Termination at the I residue is modulated by dCTP and decreases as dCTP concentration increases. A similar relationship is seen with the dCTP (1-thio) derivative, but termination is higher at given concentrations of this derivative than with dCTP. Pyrophosphate decreases general processivity in this system, but does not counteract the effect of increasing dCTP. Hill plot analysis of the dCTP effect in the inosine-containing template system gave a linear plot with Hill coefficient of 0.34, suggesting that dCTP influences termination at several steps in the polymerase reaction scheme. Substituting a methylated template base for I also increased termination, producing very strong blocks to processive synthesis. The results are consistent with a model in which termination occurs with several enzyme forms that are in equilibrium in an ordered catalytic mechanism. PMID- 2971663 TI - Human T-lymphocyte activation is associated with changes in O-glycan biosynthesis. AB - The activation of human T-lymphocytes by anti-CD3 antibodies and interleukin-2 results in a marked increase in apparent molecular weight of the major cell surface sialoglycoprotein. Both forms of the sialoglycoprotein were identified as leukosialin by a monospecific antiserum, and the differences in molecular weight were found to be due to changes in the carbohydrate structures. Our results suggest that resting T-lymphocytes express on leukosialin the disialotetrasaccharides NeuNAc alpha 2----3Gal beta 1----3(NeuNAc alpha 2--- 6)Gal-NAc-Ser/Thr, whereas activated human T-cells carry on leukosialin exclusively the more complex structures NeuNAc alpha 2----3Gal beta 1----3(NeuNAc alpha 2----3Gal beta 1----4GlcNAc beta 1----6)GalNAc-Ser/Thr. The radical shift in the biosynthetic pathway of O-glycans in activated T-lymphocytes compared to resting cells is apparently caused by a decrease of alpha 2----6 sialyltransferase activity and by the parallel dramatic stimulation of the beta 1 ---6GlcNAc-transferase. Since both enzymes compete for the same precursor substrate, the coordinate changes in their activities are most likely responsible for the complete change of the carbohydrate structures on leukosialin during the activation of human T-lymphocytes. PMID- 2971664 TI - Variation of scallop sarcoplasmic reticulum Ca2+-ATPase activity with temperature. AB - Methods for preparing native scallop sarcoplasmic reticulum vesicles, largely purified membranous scallop sarcoplasmic reticulum Ca2+-ATPase, and nonionic detergent-solubilized sarcoplasmic reticulum Ca2+-ATPase are described. The effect of a range of polyoxyethylene-based detergents on the solubilized Ca2+ ATPase was tested. Decaethylene glycol dodecyl ether (C12E10) supported the highest levels of activity, although C12E8 and C12E9 were more routinely used. Arrhenius plots of Ca2+-ATPase activity, where the assays were carried out with the same pH at all temperatures (7.4), showed a region of nonlinearity at 10 degrees C. A very similar plot was obtained when no compensation was made for pH variation with temperature. Both the break in the Arrhenius plot and the activation energies for the scallop sarcoplasmic reticulum above and below the break were very similar to those found for lobster sarcoplasmic reticulum (Madeira, V. M. C., Antunes-Madeira, M. C., and Carvalho, A. R. (1974) Biochem. Biophys. Res. Commun. 65, 997-1003). The Arrhenius plot of the scallop Ca2+ ATPase in C12E8 no longer showed the nonlinearity at 10-12 degrees C seen with the native sarcoplasmic reticulum, but instead a break now appeared at 20-21 degrees C. This is close to the Arrhenius break temperature of rabbit Ca2+-ATPase in C12E8 and of a perturbation in C12E8 (Dean, W. L. (1982) Biophys. J. 37, 56 57). PMID- 2971665 TI - Mechanism for 3,3',4',5-tetrachlorosalicylanilide-induced activation of sarcoplasmic reticulum ATPase. AB - Effects of 3,3',4',5-tetrachlorosalicylanilide (TCS), a lipophilic weak acid, on Ca2+ uptake and ATP hydrolysis by the sarcoplasmic reticulum calcium pump were characterized to obtain insight into the possible role of hydrophobic portions of the Ca2+-ATPase in the catalytic mechanism of the enzyme. TCS exhibited both the stimulatory and inhibitory effects on the calcium pump activities depending on its concentration. At optimal concentrations, it increased these activities by up to 5-fold at pH 7.0 and 6 degrees C. Analysis of partial reactions of ATP hydrolysis by the purified ATPase revealed that TCS accelerated Ca2+ release from the ADP-sensitive phosphoenzyme up to 6-fold, whereas it affected other reaction steps to a much less extent, indicating that the site of the stimulatory action of TCS is rather specific in terms of the reaction sequence. These effects of TCS became less prominent at higher temperatures, although the enzyme-TCS interactions as detected in the direct binding experiment or by measurement of quenching of protein fluorescence were not affected by a similar change in temperature. The TCS effects were also dependent on pH of the 8.0 suggested that the protonated form of TCS is responsible for both the stimulatory and inhibitory effects of the drug. These results, taken together with those obtained previously with a spin-labeled probe (Barratt, M. D., and Weaver, A. C. (1979) Biochim. Biophys. Acta 555, 337-348), may suggest that TCS stimulates the calcium pump activity through its effect on the lipid bilayer, although its direct action on hydrophobic portion(s) of the ATPase protein cannot be ruled out. PMID- 2971666 TI - Increase of the DNA strand assimilation activity of recA protein by removal of the C terminus and structure-function studies of the resulting protein fragment. AB - A proteolytic fragment of recA protein, missing about 15% of the protein at the C terminus, was found to promote assimilation of homologous single-stranded DNA into duplex DNA more efficiently than intact recA protein. This difference was not found if Escherichia coli single-stranded DNA binding protein was present. The ATPase activity of both intact recA protein and the fragment was identical. The difference in strand assimilation activity cannot be due to differences in single-stranded DNA affinity, since both the fragment and intact proteins bind to single-stranded DNA with nearly identical affinities. However, the fragment was found to bind double-stranded DNA more tightly and to aggregate more extensively than recA protein; both of these properties may be important in strand assimilation. Aggregation of the fragment was extensive in the presence of duplex DNA under the same condition where recA protein did not aggregate. The double stranded DNA binding of both recA protein and the fragment responds to nucleotide cofactors in the same manner as single-stranded DNA binding, i.e. ADP weakens and ATP gamma S strengthens the association. The missing C-terminal region of recA protein includes a very acidic region that is homologous to other single-stranded DNA binding proteins and which has been implicated in DNA binding modulation. This C-terminal region may serve a similar function in recA protein, possibly inhibiting double-stranded DNA invasion. The possible role of the enhanced double stranded DNA affinity of the fragment protein in the mechanism of strand assimilation is discussed. PMID- 2971667 TI - Distribution of isoelectric variants of the 17,000-dalton myosin light chain in mammalian smooth muscle. AB - Isoelectric focusing of purified vascular smooth muscle myosin revealed two variants of the 17,000-dalton light chain subunits. The isoelectric points of the light chain variants were determined to be 4.13 (LC17a) and 4.19 (LC17b). Tryptic peptide maps of the two species of light chain generated by reverse-phase high performance liquid chromatography disclosed small but obvious differences in peptide composition while amino acid analyses of the variants were quite similar. Two-dimensional electrophoresis of extracts from various mammalian smooth muscles revealed tissue-specific differences in the relative content of LC17a and LC17b. Vascular (aorta, carotid, and pulmonary artery) muscles and tracheal smooth muscle contained both light chain variants while smooth muscle of the gastrointestinal tract (stomach and jejunum) contained LC17a only. The actin activated Mg2+-ATPase activities of both phosphorylated and nonphosphorylated stomach (LC17b = 0) and aortic (LC17b = 40%) myosins were compared. In the presence of saturating tropomyosin, a 2-fold difference in Vmax was measured: phosphorylated, aortic, 0.119 +/- 0.009 versus stomach, 0.239 +/- 0.012 mumol of PO4 liberated/min/mg of myosin; nonphosphorylated, aortic, 0.065 +/- 0.004 versus stomach, 0.123 +/- 0.004 mumol of PO4 liberated/min/mg of myosin. In addition, the Vmax of myosin subfragment-1 ATPase from bovine aortic, pulmonary artery, and stomach myosins (LC17b contents, 40, 20, and 0%, respectively) was found to decrease in direct proportion to the LC17b content. Our results suggest that isoforms of the 17,000-dalton light chain subunits of mammalian smooth muscle myosin could play an important role in modulating actomyosin ATPase activity. PMID- 2971669 TI - Assay of isradipine and of its major metabolites in biological fluids by capillary gas chromatography and chemical ionization mass spectrometry. AB - A method is described for the determination of isradipine, a dihydropyridine calcium antagonist, and five of its metabolites in plasma and urine. The neutral compounds were extracted in toluene and analysed in a wide-bore silica capillary column. The acidic compounds were extracted in two steps, then esterified with diazomethane and assayed separately using the same column. Detection was performed by negative-ion mass spectrometry with chemical ionization. The limit of detection of isradipine was 0.04 ng/ml when the compound was determined alone and 0.7 ng/ml when its oxidized metabolite was determined simultaneously. The limits of detection of the metabolites in plasma ranged from 0.15 to 2 ng/ml. The method was successfully used in conventional pharmacokinetic studies and in a multicentre study of population pharmacokinetics. PMID- 2971668 TI - Fibronectin receptor exhibits high lateral mobility in embryonic locomoting cells but is immobile in focal contacts and fibrillar streaks in stationary cells. AB - The dynamic process of embryonic cell motility was investigated by analyzing the lateral mobility of the fibronectin receptor in various locomotory or stationary avian embryonic cells, using the technique of fluorescence recovery after photobleaching. The lateral mobility of fibronectin receptors, labeled by a monoclonal antibody, was defined by the diffusion coefficient and mobile fraction of these receptors. Even though the lateral diffusion coefficient did not vary appreciably (2 X 10(-10) cm2/S less than or equal to D less than or equal to 4 X 10(-10) cm2/S) with the locomotory state and the cell type, the mobile fraction was highly dependent on the degree of cell motility. In locomoting cells, the population of fibronectin receptors, which was uniformly distributed on the cell surface, displayed a high mobile fraction of 66 +/- 19% at 25 degrees C (82 +/- 14% at 37 degrees C). In contrast, in nonmotile cells, the population of receptors was concentrated in focal contacts and fibrillar streaks associated with microfilament bundles and, in these sites, the mobile fraction was small (16 +/- 8%). When cells were in a stage intermediate between highly motile and stationary, the population of fibronectin receptors was distributed both in focal contacts with a small mobile fraction and in a diffuse pattern with a reduced mobile fraction (33 +/- 9%) relative to the diffuse population in highly locomotory cells. The mobile fraction of the fibronectin receptor was found to be temperature dependent in locomoting but not in stationary cells. The mobile fraction could be modulated by affecting the interaction between the receptor and the substratum. The strength of this interaction could be increased by growing cells on a substratum coated with polyclonal antibodies to the receptor. This caused the mobile fraction to decrease. The interaction could be decreased by using a probe, monoclonal antibodies to the receptor known to perturb the adhesion of certain cell types which caused the mobile fraction to increase. From these results, we conclude that in locomoting embryonic cells, most fibronectin receptors can readily diffuse in the plane of the membrane. This degree of lateral mobility may be correlated to the labile adhesions to the substratum presumably required for high motility. In contrast, fibronectin receptors in stationary cells are immobilized in focal contacts and fibrillar streaks which are in close association with both extracellular and cytoskeletal structures; these stable complexes appear to provide firm anchorage to the substratum. PMID- 2971670 TI - Rapid separation of Pneumocystis carinii from lung lavage fluids. AB - A simple, rapid procedure to separate Pneumocystis carinii obtained by lavage of lungs of steroid-treated rats from rat leukocytes is described. Commercially available monoclonal antibody to rat common leukocyte determinants is used to sediment the leukocytes, resulting in supernatant fluid containing P. carinii cells virtually free of intact rat cells. PMID- 2971671 TI - Interleukin 1 stimulates its own receptor expression on human fibroblasts through the endogenous production of prostaglandin(s). AB - The regulation of interleukin 1 receptor (IL 1R) expression on human dermal fibroblasts was investigated. On exposure to IL 1 for 3 h at 37 degrees C, the capacity of fibroblasts to bind 125I-labeled human recombinant IL 1 alpha (125I IL 1 alpha) was reduced by 75%. The IL 1 binding capability of the fibroblasts was restored to control levels by 16 h after removal of unbound IL 1, and then increased to about twofold over that of control cells by 48 h. This later enhancement of IL 1 receptor expression after IL 1 treatment was abolished by indomethacin. Addition of exogenous (PGE1 and PGE2, also analogues of AMP, or forskolin increased the specific binding of 125I-IL 1 alpha to fibroblasts. Scatchard analysis indicated that PGE2 increased the number of IL 1R from approximately 1.6 X 10(3) to 5.4 X 10(3) per cell without change in the binding affinity. These data suggest that the later IL 1-induced up-regulation of IL 1R is mediated by IL 1 stimulation of endogenous prostaglandin production. The combination of PGE2 and prednisolone increased the number of IL 1R on fibroblasts in an additive manner. PMID- 2971672 TI - Platelets mediate neutrophil-dependent immune complex nephritis in the rat. AB - Neutrophils and platelets are frequently present in glomeruli in immune glomerulonephritis (GN). No role for the platelet in acute neutrophil-mediated renal injury has been defined. We investigated a neutrophil-mediated model of subendothelial immune complex GN in the rat. Rats were platelet-depleted (mean platelet less than 10,000/microliter) with goat anti-platelet IgG before induction of GN by the renal artery perfusion of concanavalin A followed by anti concanavalin A IgG. Platelet-depletion resulted in a significant reduction in albuminuria (7 +/- 2 vs. 55 +/- 10 mg/24 h) and fractional albumin excretion (0.045 +/- 0.01 vs. 0.410 +/- 0.09) compared with controls. The decrease in albuminuria was not due to differences in blood or glomerular neutrophil counts, complement, renal function, or glomerular antibody binding. Platelet-depleted rats had equivalent subendothelial deposits and glomerular endothelial cell injury but had minimal platelet infiltrates and fibrin deposition compared with controls. These studies demonstrate a role for platelets in mediating acute neutrophil-induced glomerular injury and proteinuria in this model of GN. PMID- 2971673 TI - Inactivation of human factor VIII by activated protein C. Cofactor activity of protein S and protective effect of von Willebrand factor. AB - Activated protein C (APC) acts as a potent anticoagulant enzyme by inactivating Factor V and Factor VIII. In this study, protein S was shown to increase the inactivation of purified Factor VIII by APC ninefold. The reaction rate was saturated with respect to the concentration of protein S when protein S was present in a 10-fold molar excess over APC. The heavy chain of Factor VIII was cleaved by APC and protein S did not alter the degradation pattern. Factor VIII circulates in a complex with the adhesive protein von Willebrand factor. When purified Factor VIII was recombined with von Willebrand factor, the inactivation of Factor VIII by APC proceeded at a 10-20-fold slower rate as compared with Factor VIII in the absence of von Willebrand factor. Protein S had no effect on the inactivation of the Factor VIII-von Willebrand factor complex by APC. After treatment of this complex with thrombin, however, the actions of APC and protein S towards Factor VIII were completely restored. In hemophilia A plasma, purified Factor VIII associated with endogenous von Willebrand factor, resulting in a complete protection against APC (4 nM). By mixing hemophilic plasma with plasma from a patient with severe von Willebrand's disease, we could vary the amount of von Willebrand factor. 1 U of von Willebrand factor was needed to provide protection of 1 U Factor VIII. Also in plasma from patients with the IIA-type variant of von Willebrand's disease, Factor VIII was protected. In von Willebrand's disease plasma, which was depleted of protein S, APC did not inactivate Factor VIII. These results indicate that protein S serves as a cofactor in the inactivation of Factor VIII and Factor VIIIa by APC and that von Willebrand factor can regulate the action of these two anticoagulant proteins. PMID- 2971674 TI - Gene for the rat atrial natriuretic peptide is regulated by glucocorticoids in vitro. AB - Glucocorticoids regulate the expression of the gene for atrial natriuretic peptide (ANP) in neonatal cardiocytes. Dexamethasone (Dex) increased cytoplasmic ANP mRNA levels and media ANP immunoreactivity in a dose-dependent fashion. These effects were not shared by the other classes of steroid hormones and were reversed by the glucocorticoid antagonist RU 38486. The effect on ANP mRNA levels resulted, at least in part, from enhanced transcription of the gene. Dex effected a two-fold increase in ANP gene activity assessed using a run-on transcription assay. The turnover of the ANP transcript was approximated using a standard pulse chase technique. The half-life of the ANP mRNA was 18 h in hormone-free media. In the presence of Dex this half-life increased modestly to 30 h, although the increase relative to the control did not reach statistical significance. The effect of Dex at the level of the individual myocardial cell was assessed by in situ hybridization analysis using a specific [3H]cRNA probe. These studies demonstrated a significant level of ANP expression within a subpopulation of cells in the cultures. Exposure of the cells to Dex for 24 h did not recruit additional cells into the expressing pool (27.3% cells/high power field vs. 31.3% for the control) but did increase the level of expression (i.e., grain density) within individual cells. These findings indicate that glucocorticoids stimulate expression of the ANP gene directly at the level of the myocardial cell. This results predominantly from transcriptional activation in cells already expressing the gene rather than through recruitment of previously quiescent cells. PMID- 2971675 TI - Defective expression of the 2H4 molecule after autologous mixed lymphocyte reaction activation in systemic lupus erythematosus patients. AB - Previous studies demonstrated that patients with active systemic lupus erythematosus (SLE), especially those with active renal disease, had a marked reduction in T4+2H4+ suppressor inducer cells in their peripheral blood. However, it was puzzling to find that active SLE patients without renal diseases often had normal percentages of T4+2H4+ cells. In the present study, we attempted to determine whether active SLE patients bearing normal percentages of T4+2H4+ cells had a defect in their expression of the 2H4 molecule on T4+ cells after autologous mixed lymphocyte reaction (AMLR) activation. The peripheral blood lymphocytes (PBL) from 50 SLE patients with normal percentages of T4+2H4+ cells (greater than or equal to 7% in PBL) were studied and the results were compared with those of 40 normal individuals. The density of the 2H4 molecule on T4 cells from normal controls increased during the 7-d AMLR; in contrast T4 cells from patients with SLE, especially those with active SLE, had defective expression of the 2H4 antigen after AMLR activation. Patients with inactive SLE, like normals, showed an increase in the 2H4 molecule after AMLR activation. Moreover, a strong correlation was observed between percent suppression of pokeweed mitogen (PWM) driven IgG synthesis and the density of the 2H4 antigen on AMLR-activated T4 cells. Serial analysis of patients with SLE showed that the density of the 2H4 antigen expression and the suppressor inducer activity of AMLR-activated T4 cells were inversely correlated with disease activity. Thus, defective expression of the 2H4 antigen may be an important mechanism for the failure of active SLE patients with normal percentages of T4+2H4+ cells to generate suppression. PMID- 2971676 TI - Evaluation of the antiinflammatory and phospholipase-inhibitory activity of calpactin II/lipocortin I. AB - We have examined the ability of a highly purified 38-kD phospholipase-inhibitory protein (p38) isolated from human placental membranes that is also a preferred substrate for the epidermal growth factor-urogastrone (EGF-URO) receptor/kinase, to block the release of arachidonate from zymosan-stimulated murine peritoneal macrophages in vitro and to exhibit antiinflammatory activity in a carrageenin rat paw edema test in vivo. The ability of glucocorticoids to increase the amounts of this protein in macrophage cultures was also examined. p38 represents the naturally occurring, intact, NH2-terminally blocked human placental form of the protein termed calpactin II (or lipocortin I), for which partial amino acid sequence data and a complete amino acid sequence deduced from cDNA analysis have been reported. Our data demonstrated that, whereas p38 was an effective inhibitor of pancreatic phospholipase A2 in vitro, it was unable to inhibit either the release of arachidonate from cultured zymosan-stimulated mouse peritoneal macrophages or inflammation in a rat paw edema test. At comparatively high protein concentrations, p38 enhanced either arachidonate release from intact macrophages in vitro (0.5-10 micrograms/ml) or carrageenin-induced paw swelling in vivo (2.5 or 25 micrograms per injection). Furthermore, we were unable to detect induced amounts of p38 in cultures of glucocorticoid-treated peritoneal macrophages obtained from either mice or rats. Our data indicate that the antiphospholipase activity of p38 in vitro and the ability of p38 to serve as a receptor/kinase substrate may in no way relate to the putative ability of the protein to modify eicosanoid release from macrophages in vivo, so as to modulate the inflammatory process. Our data also raise the possibility that p38 (calpactin II) may not be a true representative of the lipocortin family of glucocorticoid inducible antiinflammatory proteins, despite its ability to inhibit phospholipase A2 in vitro. PMID- 2971678 TI - Predictors of outcome in back surgery candidates. AB - This study employed a prospective design to examine the value of three demographic, three orthopaedic, and four psychometric variables as predictors of orthopaedic and rehabilitation outcome in 116 Workers' Compensation patients seriously considered for first lumbar surgery, separately for the 74 who eventually did undergo surgery, and the 42 who did not. The two groups differed only with respect to the number of Nonorganic Physical Signs, but a complex pattern of predictive relationships emerged, in which orthopaedic outcome was generally easier to predict than rehabilitation outcome. Among the demographic factors, level of English proficiency was the best predictor and correlated positively with orthopaedic outcome in both surgery and non-surgery cases and with rehabilitation outcome in non-surgery patients. The Nonorganic Physical Signs test emerged as the predictor of choice and correlated with orthopaedic outcome in both groups and with rehabilitation outcome in the non-surgery group. Scale 1 of the MMPI correlated with orthopaedic outcome in both groups and Scale 3 with both orthopaedic and rehabilitation outcome, but only in the non-surgery patients. PMID- 2971677 TI - Downregulation of interferon alpha but not gamma receptor expression in vivo in the acquired immunodeficiency syndrome. AB - Interferons (IFN) elicit antiviral and antineoplastic activities by binding to specific receptors on the cell surface. In evaluating the role of IFN as therapeutic agents in AIDS, we investigated the expression of IFN alpha and gamma receptors on peripheral blood mononuclear cells (PBM) from patients with AIDS, ARC, and heterosexual control subjects using radioiodinated IFN alpha 2 and IFN gamma. The binding characteristics of the 125I-IFN alpha and gamma to PBM were analyzed to determine receptor numbers and dissociation constants. PBM from controls expressed 498 +/- 247 IFN alpha receptor sites/cell (n = 17). However, eight patients with ARC and seven patients with AIDS had a mean number of IFN alpha receptor/cell of 286 +/- 235 (P less than 0.05) and 92 +/- 88 (P less than 0.001), respectively. This was consistent with elevated levels of serum acid labile IFN alpha and cellular 2-5A synthetase activity in patients. Treatment of PBM from the AIDS patients with exogenous IFN alpha in vitro resulted in minimal 2-5A synthetase induction in comparison to controls. In contrast, the expression of IFN gamma receptors in ARC (n = 5) and AIDS (n = 4) patients remained normal. Thus the decrease in IFN alpha receptor expression and consequent hyporesponsiveness to IFN alpha raises the question of the usefulness of IFN alpha therapy in end-stage AIDS. The normal expression of IFN gamma receptors in AIDS patients suggests that IFN gamma may prove useful in attempts to provide immune reconstitution. PMID- 2971679 TI - Chronic syphilitic meningitis resulting in superior orbital fissure syndrome and posterior fossa gumma. A report of two cases followed for 20 years. AB - Two case histories, each spanning more than 20 years, demonstrate the complexity and persistence of modern neurosyphilis. Both patients present uncommon manifestations of neurosyphilis that may be easily overlooked today: superior orbital fissure syndrome, posterior fossa gumma, and bilateral deafness. Computed tomographic scans were performed in both patients but were diagnostically nonspecific, and syphilis serology testing in both serum and cerebrospinal fluid was repeatedly misleading. Numerous mistaken diagnoses were put forward until tissue was finally available for histologic examination. Perhaps the most disturbing aspects of these two cases are the persistent failure of specific treponemal serologic testing to indicate the diagnosis, and the inability of repeated "adequate" and/or supramaximal doses of penicillin to prevent continued neurologic deterioration. We believe that chronic neurosyphilis may demand a reevaluation of current diagnostic and therapeutic practice. PMID- 2971680 TI - Neuropathologic findings in progressive supranuclear palsy. A brief review with two additional case reports. AB - Recent studies utilizing electron microscopy and immunohistochemistry have added to the understanding of the pathology of progressive supranuclear palsy. This article reviews the neuropathologic findings of progressive supranuclear palsy and includes two new case reports. Findings in the patients reported here include extensive neuronal loss with reactive gliosis in deep subcortical gray matter such as globus pallidus, subthalamic nuclei, substantia nigra, red nucleus, dentate nucleus, and inferior olives. Neurofibrillary tangles were also noted in thalamus, hypothalamus, striatum, nucleus basalis, locus ceruleus, basis pontis nuclei, midbrain, pontine, and medullary reticular formation as well as in various cranial nerve nuclei. PMID- 2971681 TI - Seesaw nystagmus. Role of visuovestibular interaction in its pathogenesis. AB - Elevation and intorsion of one eye and synchronous depression and extorsion of the other eye characterize a half cycle of seesaw nystagmus. Reversal of these movements constitutes the second half cycle, forming the "seesaw"-like movements. Based on analysis of the ocular oscillation characteristics of the cases of seesaw nystagmus reported in the literature, including the two new cases we present, we postulate that seesaw nystagmus is another type of ocular oscillation brought about by an unstable visuovestibular interaction control system. Nonavailability of retinal error signals to the inferior olivary nucleus essential for vestibuloocular reflex adaptation due to complete chiasmal dissection makes the system less stable. This system instability is further accentuated by the pursuit feedback element. The intact inferior olivary nucleus nodulus connections in seesaw nystagmus would explain the 180 degrees phase difference that distinguishes it from the midline form of oculopalatal myoclonus, where these connections are likely disrupted. PMID- 2971682 TI - Central bilateral sixth nerve palsy associated with a unilateral preganglionic Horner's syndrome. AB - Unilateral peripheral sixth nerve palsy associated with an ipsilateral postganglionic Horner's syndrome has previously been described. Isolated bilateral sixth nerve palsy due to a pontine hemorrhagic lesion, however, is a rare occurrence. We describe such a case whose only other neurological finding was a unilateral preganglionic Horner's syndrome. PMID- 2971684 TI - Acute posterior multifocal placoid pigment epitheliopathy associated with cerebral vasculitis. AB - Acute multifocal posterior placoid pigment epitheliopathy (APMPPE) is an unusual self-limited retinal disorder that has been associated with various systemic complications. To our knowledge, three prior cases associated with cerebral vasculitis have been described. This article describes a patient with APMPPE and angiographically documented cerebral vasculitis who was notable because of (a) the presence of two different cerebral ischemic events, occurring 1 month apart, and (b) the long latency (3 months) between the onset of ocular symptoms and the second cerebral ischemic event. Recognition of the association between APMPPE and cerebral vasculitis may permit early treatment of CNS involvement and prevention of morbidity. PMID- 2971683 TI - Herpes zoster ophthalmoplegia. Report of six cases. AB - Ophthalmoplegia occurs infrequently in herpes zoster ophthalmicus. The third nerve appears to be the most commonly affected and the fourth nerve the least. We describe herein the clinical course of six patients with herpes zoster ophthalmoplegia. Spontaneous recovery occurred in four patients. The pathogenesis and clinical features of this syndrome are described. PMID- 2971685 TI - Lymphoproliferative disorders. PMID- 2971686 TI - Mucocutaneous manifestations in patients infected with human immunodeficiency virus. AB - The mucocutaneous manifestations observed in 516 patients infected with human immunodeficiency virus attending an outpatient clinic for acquired immunodeficiency syndrome in Milan, Italy, from July 1985 to March 1987 are listed, and the clinicopathologic aspects of these disorders are reported. The prognostic significance of some human immunodeficiency virus-associated dermatoses is also discussed. PMID- 2971687 TI - Cutaneous manifestations of early human immunodeficiency virus exposure. AB - After demonstrating antibodies to the human immunodeficiency virus (HIV), two hundred patients were interviewed and given a complete cutaneous and mucous membrane examination. By means of the Walter Reed Staging Classification System for HIV infection, 155 patients were classified as having Walter Reed stage 1A-2A (WR1A-WR2A) infection. The prevalence of seborrheic dermatitis in this group was 36%. There were no other significant cutaneous findings in the WR1A-WR2A patient population. PMID- 2971688 TI - Dermatopharmacology: drugs around the corner. AB - This article describes impending advances in drug treatment of skin diseases. We have included the following categories: (1) interesting new drugs for which there are reasonable prospects of eventual licensing for skin indications, (2) important novel compounds whose progress through licensing procedures, although incomplete at the time of writing, is advanced, and (3) new indications or improved regimens that have recently emerged for established drugs. PMID- 2971689 TI - Successful treatment of hidradenitis suppurativa with acitretin. PMID- 2971690 TI - Use of oral and topical agents for acne in pregnancy. AB - Dermatologists frequently are consulted by a pregnant patient or a woman of childbearing age who desires acne therapy. Because there are no published studies in which women took acne medications throughout pregnancy, information about safety must be obtained indirectly from studies in which the agents were taken for another indication during some portion of pregnancy. Oral tetracycline is associated with maternal liver toxicity and deciduous tooth staining in the infant, and tetracycline occasionally has been associated with other congenital anomalies. Maternal isotretinoin ingestion is associated with major craniofacial and cardiac deformities, as well as other congenital anomalies. Erythromycin, however, appears to be safe. Topical acne medications never have been implicated as a cause of fetal deformities in human beings. Dermatologists should be aware of potential toxic and teratogenic effects of acne medicines before prescribing them to women of childbearing age. Prompt reporting of adverse effects is encouraged. PMID- 2971691 TI - Cutaneous manifestations of human immunodeficiency virus in Lusaka, Zambia. AB - Of the 1124 human immunodeficiency virus (HIV)-infected patients studied, one or more cutaneous lesions were found in 113 (98.3%) of the 115 patients with acquired immunodeficiency syndrome (AIDS) and in 541 (53.6%) of 1009 patients with AIDS-related complex (ARC). Kaposi's sarcoma, multidermatomal, necrotic herpes zoster, and pruritic maculopapular rashes are common cutaneous manifestations of AIDS and its related complex (ARC) in Zambia. The maculopapular rash results from a lymphoplasmacytic angiitis in the dermis, possibly in response to the presence of HIV in the dermis. Candidiasis, severe genital herpes, extensive molluscum contagiosum, and tinea corporis were less frequent and usually refractory to treatment. Drug reactions are also frequent in Zambians with AIDS. In seven patients given streptomycin, thiacetazone, and rifampicin for treatment of pulmonary tuberculosis, Stevens-Johnson syndrome occurred shortly after therapy was begun, and two died despite high-dose prednisone and discontinuance of tuberculosis therapy. Extensive seborrheic dermatitis refractory to topical fluorinated corticosteroids is also an associated condition in AIDS patients who have pulmonary tuberculosis. PMID- 2971692 TI - Acitretin versus etretinate in psoriasis. Clinical and pharmacokinetic results of a German multicenter study. AB - 175 patients with severe psoriasis of different types were treated with 10, 25, or 50 mg acitretin and compared with patients receiving 50 mg etretinate over a period of 8 weeks in a randomized, double-blind multicenter study in the Federal Republic of Germany. Plasma concentrations of etretinate and its metabolite acitretin were measured during therapy and also 3 weeks after cessation of treatment. After 4 weeks of treatment, a trend toward clinical improvement was shown in all groups with increasing dosage. Those groups receiving the lower acitretin doses (i.e., 10 and 25 mg/day) had more dropouts than the groups taking 50 mg acitretin or 50 mg etretinate. Complete remissions before the end of therapy occurred only among those receiving higher doses. Enlargement of psoriatic lesions, however, could be observed during treatment with both retinoids, despite improvement of other parameters, as measured by psoriasis area and severity index (PASI) and psoriasis severity index (PSI). After 8 weeks, a significant improvement was calculated by the PASI score and by a newly defined, corrected PASI score for all four dose regimens compared with baseline levels. A greater than 50% PSI score improvement was seen in 50% of patients treated with 10 mg acitretin, 40.5% with 25 mg acitretin, 53.8% with 50 mg acitretin, and 61.1% with 50 mg etretinate. No statistical differences were found among these groups at any time during the 8-week period. No new or unexpected side effects occurred during acitretin treatment. Moreover, cholesterol levels did not significantly change. Three weeks after cessation of drug administration, the plasma concentrations of acitretin were below the sensitivity level of the assay, whereas etretinate was still quantifiable. It is interesting that acitretin plasma concentrations during therapy with 50 mg acitretin were markedly lower (means = 18 ng/ml) than were acitretin levels during treatment with 50 mg etretinate (means = 36 ng/ml). PMID- 2971693 TI - Comparison of the stability of topical isotretinoin and topical tretinoin and their efficacy in acne. AB - After 4 hours of exposure to incandescent light, 80% of 0.05% topical isotretinoin and 60% of 0.05% topical tretinoin preparations remained in their original form. In contrast, after 2 hours of exposure to fluorescent light only 25% of topical tretinoin and possibly 60% of topical isotretinoin remained in their original forms. Longer exposure to fluorescent light did not result in further breakdown, and the final breakdown of both preparations was similar. A 12 week, double-blind clinical trial comparing isotretinoin (0.05%) with tretinoin (0.05%) applied topically to patients with moderate acne was carried out. Both preparations caused significant reductions in papules and pustules. However, neither treatment was significantly superior to the other in the reduction of acne lesions. This may be because both preparations are rapidly broken down to similar products when exposed to fluorescent light. PMID- 2971695 TI - Pustular eruption following administration of cefazolin: a second case report. PMID- 2971694 TI - Milk tester's dermatitis. AB - Hand dermatitis is a frequent problem among workers in milk testing laboratories. An epidemiologic study was conducted at the Pennsylvania Dairy Herd Improvement Association Milk Testing Laboratory, where more than 300,000 milk samples are examined monthly for protein, butterfat, and "somatic" cells. These samples are preserved with potassium dichromate for transport from the farm to the laboratory. A survey of the laboratory was conducted and workers were interviewed. Eight of 16 subjects reported a history of occupationally exacerbated hand dermatitis. Three of 16 subjects had positive patch test results to potassium dichromate. Two of 15 subjects who underwent patch testing to milk preserved with potassium dichromate had positive reactions. None reacted to milk alone, bronopol, or Kathon CG. Two workers are receiving workers' compensation because of severe allergic contact dermatitis of the hands to potassium dichromate. We conclude that milk testing laboratory workers are at substantial risk for acquiring allergic contact dermatitis from milk preserved with potassium dichromate. PMID- 2971696 TI - Homology of glucosyltransferase gene and protein sequences from Streptococcus sobrinus and Streptococcus mutans. AB - The sequences of glucosyltransferase genes from Streptococcus sobrinus (gtfI) and Streptococcus mutans (gftB) were compared and show a high degree of homology. There is a 57.7% homology of nucleotides in the genes and a 56.7% homology of amino acids in the deduced protein sequences. The G + C content for the protein coding region is 43.6% for S. sobrinus and 41.2% for S. mutans. Internal repeating sequences present in both proteins exhibit some difference in sequence pattern. PMID- 2971698 TI - The antibiotic susceptibility of Propionibacterium acnes: a 15-year bacteriological study and retrospective evaluation. PMID- 2971697 TI - Cross-sectional analysis of serum antibody to oral streptococcal antigens in children. AB - Antibodies to S. salivarius, S. sanguis, and S. mutans cells and to glucosyltransferases (GTF) prepared from these micro-organisms were measured in the sera of 133 infants and children by enzyme-linked immunosorbent assay (ELISA). IgG antibody activity to each cell type and GTP was present at birth (presumably derived from maternal transfer) and declined significantly thereafter. IgG antibody levels to S. salivarius and S. sanguis were next detected in young children (2 to less than 3 yr group). However, an increase in IgG antibody to S. mutans cells was not seen until children were older (4 to less than 8 yr group), possibly reflecting the later colonization of this organism. In contrast, IgG antibody to GTF of all three streptococcal species remained at low levels throughout the first four years of life. IgG antibody to S. mutans GTF was then the first to appear (4 to less than 8 yr group). Serum IgA antibodies to all GTFs were not detected until after this time. Fifteen sera were used to develop IgG immunoblots with the GTF antigens. Some positive sera (7/12) demonstrated reaction(s) with GTF from each of the three streptococcal species. Individual sera showed IgG antibody bonds to GTF from several serotypes of the mutans streptococci. No immunoblot reaction was observed with GTF and sera (3) from the four-to-seven-year and younger age groups. These results indicate the presence of serum antibody to bacteria and bacterial products associated with plaque formation very early in life and during and after the pre-adolescent years. The potential exists for this serum antibody to modulate bacterial colonization or accumulation in the oral cavity. PMID- 2971699 TI - Comparison of complications during percutaneous transluminal coronary angioplasty from 1977 to 1981 and from 1985 to 1986: the National Heart, Lung, and Blood Institute Percutaneous Transluminal Coronary Angioplasty Registry. AB - Because the effects of changing technology in percutaneous transluminal coronary angioplasty, increased operator experience and use of the procedure in patients with extensive disease are unknown in regard to complication patterns, the initial 1977-1981 cohort and the recent 1985-1986 cohort of the National Heart, Lung, and Blood Institute Percutaneous Transluminal Coronary Angioplasty Registry were analyzed with respect to complications. Compared with the initial cohort of 1,155 patients, the 1,801 new cohort patients were older and had an increased prevalence of multivessel coronary artery disease, depressed left ventricular function and prior infarction. Overall complication rates in the recent cohort were either unchanged or decreased from the rates in the initial cohort despite a higher risk patient population. The most significant decreases were in the incidence of coronary spasm (p less than 0.001) and the need for emergency coronary bypass surgery (p less than 0.01). Overall in-hospital mortality was low but was dependent on the extent of vessel disease--0.2% for single vessel disease, 0.9% for double vessel disease and 2.2% for triple vessel disease (p less than 0.001 for linear trend). Acute coronary complications of branch occlusion, dissection or abrupt closure were associated with increased rates of death, nonfatal infarction or need for emergency surgery. Factors showing a multivariate association with increased mortality included a history of congestive heart failure (p less than 0.001), age greater than or equal to 65 years (p less than 0.01), triple vessel or left main coronary artery disease (p less than 0.05), female gender (p less than 0.05) and new onset angina.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971700 TI - Posterior myocardial infarction revisited. PMID- 2971701 TI - Reversal of chronic ischemic myocardial dysfunction after transluminal coronary angioplasty. AB - From a cohort of patients referred for elective transluminal coronary angioplasty, a subset of patients was evaluated to determine whether revascularization using coronary angioplasty could salvage chronically ischemic myocardium. Reversible chronic ischemic left ventricular dysfunction was identified by a severe wall motion abnormality at rest and at least one of the following: 1) persistent angina pectoris; 2) postextrasystolic ventricular contraction potentiation of motion in the asynergic zone on baseline ventriculogram; and 3) thallium-201 uptake in the asynergic zone. Twelve patients were identified as having reversible chronic ischemia and underwent coronary angioplasty. Their mean age was 63 +/- 11 years and duration of symptoms 8.3 +/- 9.7 weeks. Immediate pre- and postangioplasty left ventriculograms were obtained. Regional wall motion was analyzed using a radial axis model, and global ejection fraction was calculated. After angioplasty, tension development (heart rate systolic pressure product) increased in the absence of an increase in left ventricular end-diastolic pressure. Global ejection fraction increased from 46 +/ 20 to 62 +/- 19% (p less than 0.005). The percent of left ventricular diastolic perimeter showing asynergy decreased from 29 +/- 11 to 10 +/- 13% (p less than 0.005). During follow-up ranging from 6 to 51 months, sudden death occurred in one patient who had had no improvement in wall motion after angioplasty, repeat angioplasty was performed in three patients and eight patients remained asymptomatic. Application of easily obtainable clinical data identifies a subset of patients with chronically ischemic myocardium. Coronary angioplasty in such patients is useful in salvaging hibernating myocardium. PMID- 2971702 TI - Stress thallium-201 imaging after coronary angioplasty predicts restenosis and recurrent symptoms. AB - The predictive accuracy of thallium imaging for the diagnosis of restenosis after angioplasty was evaluated in 121 patients who had undergone a successful procedure. Patients were evaluated three times over a 1 year follow-up period for symptoms, electrocardiographic (ECG) changes during exercise and the presence of reversible ischemia on exercise thallium imaging. At initial evaluation (4 to 6 weeks after angioplasty), 104 patients (86%) were asymptomatic. Of the 17 patients with chest pain symptoms, 9 had reversible ischemia on thallium imaging and all had restenosis. Of the 104 asymptomatic patients, 26 (25%) had a positive thallium scan (reversible ischemia) and this sign identified a high risk group. Evidence of restenosis was present by 6 months in 22 (85%) and by 1 year in 25 (96%) of these 26 patients. The largest group of patients manifested symptoms by the second evaluation (between 3 and 6 months after angioplasty). Of 28 patients with symptoms and a positive thallium scan at this evaluation, 26 had restenosis; on initial evaluation, 22 of these 26 patients had no symptoms but had had a positive thallium study. Ten of the 65 patients in the asymptomatic group had a positive thallium scan, but the scans of this group were not as predictive for recurrent symptoms as were those of the same group at initial evaluation. The 74 patients who had a negative thallium scan at 3 to 6 months with or without associated symptoms had a low likelihood of developing restenosis or symptoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971703 TI - Application of percutaneous transluminal coronary angioplasty to the internal mammary artery graft. AB - Between June 1982 and August 1987, 45 patients underwent percutaneous transluminal coronary angioplasty within the internal mammary artery graft (group 1) or had coronary angioplasty performed beyond the graft insertion with the internal mammary artery used as a conduit (group 2). Previous coronary artery bypass surgery was performed at a mean of 29.8 months (range 2 to 199) in group 1 and 51.3 months (range 2 to 134) in group 2. Sixteen (62%) of 26 patients in group 1 and 17 (71%) of 24 patients in group 2 had multivessel angioplasty; multilesion angioplasty was performed in 20 patients (77%) in group 1 and in 19 patients (79%) in group 2. Within group 1, 12 (37.5%) of 32 lesions were in the body of the internal mammary artery graft and 20 lesions (62.5%) occurred at the distal anastomosis. Angioplasty was successful in 30 (94%) of 32 attempts in group 1 and in 25 (96%) of 26 attempts in group 2. Procedure-related complications were limited to emergent bypass surgery in one patient in group 2. At a mean follow-up period of 12.7 months in group 1 and 18.2 months in group 2, 39 (90%) of the 45 patients had no or only mild angina. There were two late cardiac deaths (mortality rate 4.9%) in the 41 patients with successful angioplasty. The results of this study demonstrate that percutaneous transluminal coronary angioplasty of internal mammary artery grafts combined with multilesion angioplasty is technically feasible, can be performed with a high primary success rate and a low incidence of complications and achieves sustained clinical improvement in the majority of patients. PMID- 2971704 TI - Increased afterload aggravates infarct expansion after acute myocardial infarction. AB - After acute transmural myocardial infarction, the heart may undergo major remodeling characterized by thinning and dilation of the infarct zone and overall enlargement of the heart. The effect of increased left ventricular pressure on infarct expansion and the extent to which it alters postinfarction remodeling were studied in a rat model. Rats with either aortic banding or a sham operation and a survival period of 3 weeks were further randomized to sham thoracotomy or left coronary ligation. Surviving rats were killed 7 days later and the hearts were fixed in diastole for morphologic analysis. Hearts with aortic banding had a mean peak to peak gradient of 20.7 +/- 4.9 mm Hg across the aortic band at death and a significantly thicker heart than that of the comparison group without an aortic band. Infarct size, as a percent of total left ventricular mass, at the time of death was less in the group with aortic banding, yet infarct expansion was more marked. However, when original infarct size was estimated taking into account the effects of aortic banding, scar formation, infarct expansion and infarct-induced hypertrophy, it was found to be similar in both infarct groups (45.50 +/- 4.2 versus 47.90 +/- 3.1%). Infarct expansion, as measured by cavity dilation and infarct thinning, occurred in both infarct groups but was greater in the group with aortic banding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971705 TI - Doppler flow velocity mapping in an in vitro model of the normal pulmonary artery. AB - Pulsed Doppler pulmonary artery velocity measurements are useful in evaluating a number of cardiac conditions including pulmonary hypertension, pulmonary stenosis and insufficiency, intracardiac shunts and other congenital abnormalities. However, variations in sample location relative to the arterial wall and valve have been thought to affect pulmonary artery velocity and acceleration measurements clinically. Therefore, pulsed Doppler and color flow mapping were performed in a pulsatile flow apparatus connected to a glass or Plexiglas model of the main pulmonary artery and its bifurcation, which contained a Hancock 29 mm pericardial tissue valve (5.35 cm2 orifice). Doppler sample volumes were placed at four sites: 1) at the pulmonary valve leaflet tips, centrally; 2) 2 cm distal to the leaflet tips, centrally; 3) 2 cm distal but laterally near the pulmonary artery wall; and 4) at the pulmonary artery bifurcation, centrally. Doppler peak flow velocity and acceleration time were measured. There was no difference between sites 1 and 2 in peak flow velocity or acceleration time. At site 3, peak flow velocity and acceleration time were both less than at site 1 (mean +/- SD, 85 +/- 44 versus 105 +/- 39 cm/s, p less than 0.005, and 162 +/- 65 versus 188 +/ 46 ms, p less than 0.03, respectively). Moreover, the pulmonary artery velocity contour at site 3 exhibited increased spectral dispersion and notching and increased variance on the color spectrum. At site 4, peak flow velocity was less than at site 1 (85 +/- 31 versus 105 +/- 39 cm/s, p less than 0.005), whereas pulmonary artery acceleration time was not significantly different. In this model, Doppler pulmonary artery flow velocity was best recorded within 2 cm of the valve and in the center of the vessel. Similar studies should be performed in the human pulmonary artery to standardize the recording technique and sample sites for Doppler measurements of velocity and acceleration. PMID- 2971706 TI - Intracoronary verapamil for reversal of refractory coronary vasospasm during percutaneous transluminal coronary angioplasty. AB - Coronary artery spasm unresponsive to intracoronary nitroglycerin was observed in eight patients undergoing percutaneous transluminal coronary angioplasty for unstable ischemic symptoms (unstable angina or recent nontransmural infarction, or both). All patients manifested eccentric lesions angiographically with the right coronary artery involved in four, circumflex artery in two and left anterior descending in two. Severe coronary spasm was documented angiographically in all patients after angioplasty and resulted in symptomatic and electrocardiographic evidence of ischemia. Multiple sites of spasm were present in the dilated vessel in three patients. Coronary artery spasm persisted despite the infusion of large doses of intracoronary nitroglycerin (200 to 2,000 micrograms, mean 850 micrograms) over 10 min. Administration of intracoronary verapamil (1 to 1.5 mg over 10 min) resulted in complete relief of spasm with restoration of brisk anterograde flow in all patients. These findings suggest that intracoronary verapamil may be a useful agent for the relief of coronary spasm occurring in the setting of coronary angioplasty. PMID- 2971707 TI - Longitudinal study of alveolitis in hypersensitivity pneumonitis patients: an immunologic evaluation. AB - Cells recovered from bronchoalveolar lavage were studied, both from a phenotypic and functional point of view, in 18 patients with hypersensitivity pneumonitis (HP) during a prolonged follow-up. A series of monoclonal antibodies against different lymphocyte subpopulations, including T cells, T cell subsets, and natural killer (NK) cells have been used. In some cases, an immunohistologic analysis of lung tissue sections has also been performed. The NK activity has been evaluated with regard to the in vitro function. At the time of the first evaluation, a high number of CD8+ cells with an imbalance of CD4/CD8 ratio had been demonstrated in patients with HP. Consecutive bronchoalveolar lavage evaluations demonstrated a persistent increase of CD8+ cells and a reversal of CD4/CD8 ratio in patients who continued to be regularly exposed to etiologic antigens at work (W+). In the same cases, a persistent increase of NK cells was demonstrated. Cytotoxic cells demonstrated a persistently enhanced in vitro lytic function during the follow-up, even though there appeared to be a trend toward the normal range. Patients who continued to live in agricultural environments but were not further exposed to specific antigens at work (W-) exhibited a recovery of CD4+ cells, a decrease in CD8+ cells, and an increase of CD4/CD8 ratio to the normal range 6 months after the first observation. Immunohistologic analysis, performed at the time of the first evaluation, demonstrated a diffuse infiltration of lung parenchyma by CD8+ cells, both in W+ and W- patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971708 TI - Effect of vasoactive intestinal peptide (VIP) on propranolol-induced bronchoconstriction. AB - There is now considerable evidence in favor of vasoactive intestinal peptide (VIP) as a neurotransmitter of nonadrenergic noncholinergic nerves in the airways. The purpose of our study was to evaluate the influence of inhaled VIP on bronchomotor tone after a beta-adrenergic- and cholinergic-receptor blockage. The study was performed in six patients with asthma in 4 days. On the first day, a propranolol provocative dose producing a 20% change in FEV1 (PD20) was determined from the individual semilogarithmic dose-response curve. On the other days, the propranolol challenge was performed after inhalation of ipratropium bromide (40 micrograms), VIP (70 micrograms), and both drugs in randomized double-blind order. Statistical analysis was performed by two-way analysis of variance. The results demonstrated that mean propranolol PD20 was 0.14 mg (geometric mean + SD = 1.22). Ipratropium bromide administration, like VIP administration, significantly raised the PD20 value. The administration of both drugs elicited a further remarkable increase of mean propranolol PD20. The results demonstrated that inhaled VIP influences bronchomotor tone and that this effect is independent of the cholinergic blockage. PMID- 2971709 TI - Proportional hazards analysis of risk factors for coronary heart disease in individuals aged 65 or older. The Framingham Heart Study. AB - Risk factors for coronary heart disease were examined in 2,501 individuals in the Framingham Heart Study who survived to the age of 65 without evidence of coronary artery disease. We used a proportional hazards (Cox) analysis that examined risk factors over time and included events through the 16th biennial examination. The independently significant multivariate correlates of the development of coronary heart disease after the age of 65 were sex (incidence rate ratio [RR] for males of 1.7 [95% confidence interval of 1.4,2.0]), left ventricular hypertrophy (RR = 2.4 [1.7,3.5]), systolic blood pressure (RR = 2.2 [1.4,3.3] for systolic blood pressure of 160 mmHg or higher as compared with less than 120 mmHg), casual blood glucose (RR = 2.2 [1.5,3.4] for 175 mg/dL or more as compared with less than 90 mg/dL), Metropolitan relative weight from examination 1 (RR = 1.3 [1.0,1.6] for those 130% or more of ideal weight compared with those less than 110% of ideal), and total serum cholesterol (RR = 1.8 [1.3,2.5] for cholesterol in the 90th percentile or higher compared with less than 200 mg/dL). Controlling for all these risk factors, those smoking 20 cigarettes a day or more were at slightly increased risk of coronary heart disease (RR = 1.2 [.9,1.6]) compared with nonsmokers. These analyses suggest that older persons share the same risk factors for coronary artery disease that are significant in younger populations. PMID- 2971710 TI - Heart and lung hypertrophy, changes in blood volume, hematocrit and plasma renin activity in rats chronically exposed to increasing carbon monoxide concentrations. AB - Changes in blood volume, heart and lung mass and composition and plasma renin activity were examined in two strains of male albino rats inhaling incrementally increasing concentrations of CO: 250 ppm for 17 days, 500 ppm for 13-14 days, 750 ppm for 10 days, and 1300 ppm for 10 days. Blood volume increased 86% and erythrocyte mass increased 212% at 1300 ppm, while plasma volume was unchanged or decreased slightly. 'Real' hematocrit (hematocrit corrected for plasma trapping) increased from 50% in controls to a peak of 75% at 1300 ppm CO. Wet weight of right ventricle (RV) and combined right and left atria (2A) increased linearly with CO concentration, paralleling changes in blood volume; while real hematocrit increased non-linearly. Left ventricle + interventricular septum (LV + S) wet weight increased less than RV, but more in Sprague-Dawley than in Wistar rats. Plots of RV, LV + S and 2A weight vs real hematocrit showed sharp upward inflections at real hematocrit 65%, suggesting a possible role of increased viscosity in CO cardiomegaly at the higher hematocrit. Assymetric septal hypertrophy was not present. Lung weight increased with CO concentration, but was not due to increased lung blood volume or edema. Hydroxyproline measurements on heart and lung failed to show increased collagen content. Plasma renin activity measured by radioimmunoassay was depressed at 500 ppm, but was normal at 750 and 1300 ppm CO. Neither heart, lung, liver nor plasma renin measurements suggest congestive heart failure, supporting previous studies in the rat with chronic carboxyhemoglobinemia, in spite of enormously increased blood volume, hematocrit and heart weight. PMID- 2971711 TI - Quantitative and functional change in T cells of primiparous mice following injection of benzo(a)pyrene at the second trimester of pregnancy. AB - Progeny from benzo(a)pyrene (BP) exposed (150 micrograms/g body weight) primiparous mothers, injected during the second trimester of pregnancy, are severely compromised immunologically. In view of maternal-fetal associations, we studied, during pregnancy and postpartum, the quantitative and functional status of the maternal T cell population in the thymus and/or spleen. In the thymus, there is an exacerbated depression in the amounts of thymocytes, (theta +, Lyt 1+, Lyt 2+ cells) during pregnancy relative to the corn oil-injected controls, which is sustained postpartum. In the spleen, while there are inconsistencies in the level of theta + cells, The Lyt 1+ are depressed postpartum relative to virgins, but the Lyt 2+ are enhanced during pregnancy and postpartum, reaching levels greater than 700-fold of controls. In controls, while the number of Lyt 1+ cells was higher than BP-exposed mice or virgins during pregnancy, they virtually disappeared postpartum. A similar but reversed image is mirrored by the Lyt 2+ cells, i.e., they were virtually absent during pregnancy but increased postpartum. Splenic allogeneic and syngeneic mixed lymphocyte responses were subnormal. The data show that BP disrupts the maternal T cell repertoire, leading to an accumulation of Lyt 1-2+ cells, and suggest that splenic disruption may be due to changes in the differentiation potential of T precursor cells. These changes not only are most likely to affect maternal defenses, but also may have a direct bearing on the establishment of progeny immune status. PMID- 2971712 TI - Hepatitis B and dialysis patients. PMID- 2971713 TI - Are we trading disability for death? PMID- 2971714 TI - Relationship between plasma atrial natriuretic peptide and left atrial and left ventricular involvement in essential hypertension. AB - In order to investigate the role of cardiac hypertrophy in atrial natriuretic peptide (ANP) secretion in patients with essential hypertension, plasma levels of ANP were measured after overnight rest in 36 patients with untreated hypertension and in 31 normotensive controls. In the hypertensive subjects, plasma levels were correlated with left ventricular (LV) and left atrial abnormalities detected by chest X-ray, electrocardiogram (ECG) and M-mode echocardiography. Plasma ANP levels in patients with hypertension averaged 146 +/- 27 pg/ml compared to 46 +/- 7 pg/ml in the normotensive subjects (P less than 0.001). In patients with hypertension a significant correlation was found between ANP and supine systolic blood pressure (r = 0.54, P less than 0.001) and between ANP and diastolic blood pressure (r = 0.38, P less than 0.05). Furthermore, plasma ANP levels were correlated with total heart volume (r = 0.68, P less than 0.01), LV mass (r = 0.525, P less than 0.001), LV posterior wall thickness (r = 0.39, P less than 0.05), Sokolow-Lyon index (r = 0.721, P less than 0.001) and end-diastolic diameter of the left atrium (r = 0.334, P less than 0.05). The results suggest a contribution of LV and left atrial abnormalities to ANP secretion in essential hypertension. PMID- 2971715 TI - Clonal analysis of mechanisms of murine T helper cell collaboration with effector cells of macrophage lineage. AB - The molecular basis and genetic restrictions of collaboration between Th cells and macrophages (Mo) and the numbers of types of collaboration in the Ag-specific cellular immune response were analyzed. Using the response of cloned Ag-specific T cells we examined the mechanisms of induction of the macrophage procoagulant response. Two generic types of collaboration were identified. One was mediated by the lymphokine monocyte procoagulant inducing factor (MPIF) and the second mechanism was by apparent contact collaboration. The lymphokine MPIF was produced by T cells and cloned CD4+ T cells after specific Ag stimulation. Cloned CD8+ cells, most of which also exhibited cytolytic activity, produced little MPIF. There was no evident restriction of the response of Mo of different MHC or background genes. In the second collaborative pathway a subset of CD4+ cloned Th cells were able to directly collaborate by an apparent contact mechanism with Mo for the procoagulant response. There was no correlation of this latter capacity with MPIF production. In addition abrogation of protein synthesis and lymphokine production by Ag-driven clones did not abrogate the direct cell association type of collaboration. Both forms of collaboration were equally efficient across MHC incompatibility barriers and different genetic background. We conclude that there are two independent and parallel Th:Mo collaborative pathways for Ag-driven responses in this limb of the cellular immune response, i.e., a MPIF lymphokine pathway and a contact pathway, and that there are quantitative and qualitative clonal differences in the use of these two pathways. We suggest that the existence of multiple parallel pathways for cellular collaboration may occur more widely in the Th:Mo limb of the immune response in respect to other Mo effector molecules and should be explored to understand the orchestration of this limb of the immune response. PMID- 2971716 TI - Expression, distribution and specificity of Fc receptors for IgM on murine B cells. AB - Subpopulations of normal adult murine splenic B cells and a panel of murine B cell tumors were examined for their ability to bind murine IgM specifically. By using two-color flow cytometric analyses, we have demonstrated that 90 to 95% of surface (s)IgD+ B cells express surface membrane receptors for IgM (Fc mu R). The binding of pentameric murine IgM to splenocyte Fc mu R was IgM-specific since it was totally inhibited by other polymeric IgM proteins, but not by Ig of other H chain classes or by mAb specific for the murine IgG or IgE FcR. Binding of IgM to splenic cells was saturable. Fc mu R were co-expressed with the Fc gamma R as well as the Fc epsilon R on the majority of splenic B cells. Minor populations of splenic mononuclear cells expressed only an Fc mu R, Fc gamma R or Fc epsilon R. In a survey of B tumor cell lines representing different stages of B cell development, we observed that the Fc mu R was expressed on pre-B cell lines and that Fc mu R detection was maximal on immature B cell lines that expressed sIgM and low amounts of sIgD and Ia. Fc mu R were not detected on cell lines that had switched from sIgM to the expression of another sIg, or on plasmacytomas and hybridomas. The studies with normal splenocytes establish that the majority of sIgD+ B lymphocytes in adult BALB/c mice express surface membrane receptors that specifically bind IgM. The studies with B lineage tumor cells suggest that the expression of Fc mu R on B cells is developmentally regulated and that the pattern of expression exhibited by Fc mu R during B cell ontogeny differs from the patterns that have been previously found for IgG and IgE FcR. These observations raise the possibility that Fc mu R might have a functional significance in some aspect of B cell maturation and activation. By using a family of IgM H chain constant region domain deletional mutants, we have further demonstrated that, like the T cell Fc mu R, the B cell Fc mu R also requires a C mu 3 domain for binding to occur, raising the possibility that the T and B cell Fc mu R in mice may be structurally related molecules. PMID- 2971717 TI - Cloned and expressed human Fc receptor for IgG mediates anti-CD3-dependent lymphoproliferation. AB - We have utilized gene transfer experiments to investigate the role of a human monocyte receptor for IgG (Fc gamma RII) in mouse IgG1 anti-CD3 (Leu 4)-induced lymphoproliferation in vitro. Mouse Ltk- cells expressing human Fc gamma RII or a mutant of Fc gamma RII lacking the entire cytoplasmic domain of the receptor mediate anti-CD3-induced lymphoproliferation in cultures of adherent cell depleted human PBMC. Expression of an Fc gamma RII mutant lacking transmembrane and cytoplasmic domains (soluble Fc gamma RII) in COS7 cells yielded a secreted receptor which retained affinity for IgG, even in the absence of the mutant receptor's N-linked oligosaccharides. Soluble Fc gamma RII inhibits rosette formation by human IgG-sensitized RBC and the Fc gamma RII-bearing cell line K562, but does not sitmulate anti-CD3-induced lymphoproliferation under the conditions tested. PMID- 2971718 TI - Human recombinant IL-4 induces activated B lymphocytes to produce IgG and IgM. AB - In this report, we describe a novel biologic activity of IL-4 namely, its ability to induce activated human B cells to produce IgM. Staphylococcus aureus Cowan I activated blasts prepared from high density tonsil B cells were found to secrete IgG and IgM, but no IgE, when cultured in the presence of rIL-4. The differentiating activity of rIL-4 was totally blocked by a neutralizing anti-IL-4 antiserum, therefore demonstrating that the IgG/IgM-inducing activity of rIL-4 was an intrinsic property of IL-4. rIL-4 was only minimally inducing Ig production of blasts prepared from low density B cells, whereas it induced B cell blasts prepared from high density B cells to secrete a high amount of Ig. Delayed additions of the neutralizing anti-IL-4 antiserum demonstrated that a 48-h contact between IL-4 and B cell blasts was required for optimal Ig production. The IL-4-mediated IgG and IgM production was neither suppressed by IFN-gamma nor by anti-CD23 mAb 25, whereas these agents have been shown earlier to inhibit IgE production of enriched B cells cultured in the presence of IL-4. These data indicate that the IgG/IgM-inducing activity of IL-4 is not regulated like the IL 4-induced IgE production by enriched B cells. PMID- 2971719 TI - Spontaneous IFN-beta production. A common feature of natural suppressor systems. AB - Natural suppression has been described in several immunologic systems in which splenocytes are not only incapable of generating an immune response, but are also able to non-specifically suppress normal splenocyte reactivity. Natural suppressor cells exist in such transitory immune deficiency states as graft vs host disease, irradiation recovery, and during neonatal development. We have found that when splenocytes from each of these three systems were placed in culture, IFN was produced spontaneously without stimulus. This spontaneous IFN production was augmented by the addition of IL-2 or granulocyte-macrophage-CSF to cell cultures. However, these lymphokines induced little or no IFN from normal splenocytes. This unusual IFN production is especially interesting since all samples have been typed to be IFN-beta. Additionally, greater IFN-beta was produced by these spleen cells during active suppression of the MLR of normal spleen cells. In fact, antibody to IFN-beta was shown to partially reverse the natural suppression mediated by graft vs host disease splenocytes in the MLR. Thus, IFN-beta production correlates extremely well with natural suppressor states. As mice begin to recover normal immune responses in all three systems, IFN-beta ceases to be produced spontaneously in culture. These findings establish a previously undescribed IFN-beta-producing splenic phenotype and suggest that IFN-beta may contribute to the immunoregulation of natural suppressor systems. PMID- 2971720 TI - Expression of C gamma 4 T cell receptors and lack of isotype exclusion by dendritic epidermal T cell lines. AB - Although four murine C gamma gene segments (C gamma 1, 2, 3, and 4) are known to exist, the large majority of expressed gamma-chains have been shown to be of the C gamma 1 isotype and no evidence exists for the expression of more than one receptor by gamma delta TCR-bearing cells. We investigated the nature of the TCR expressed on a number of murine dendritic epidermal T cell-derived cell lines by using both Northern blot and immunoprecipitation analyses. One of these CD3+ cell lines (T195) expresses C gamma 4, V gamma 1, and delta mRNA, and its CD3 associated TCR complex can be precipitated by both anti-C gamma 4 and anti-delta sera, indicating that this receptor is a C gamma 4/delta heterodimer. Furthermore, we show that two cell lines (Y245, Y93) express two distinct TCR gamma-chains, one derived from the C gamma 4 locus, whereas the second gamma chain is probably derived from the C gamma 2 locus. Together with the previous demonstration of C gamma 1/delta TCR on a number of dendritic epidermal T cell lines (DETC), these results indicate that such DETC are capable of expressing a variety of gamma delta TCR and that, in some DETC, isotype exclusion of gamma chain expression does not occur. PMID- 2971721 TI - Possible role of human lymphocyte receptor for IgE (CD23) or its soluble fragments in the in vitro synthesis of human IgE. AB - The present study demonstrates that human rIL-4 is capable of inducing the secretion of IgE by PBMC. At a concentration of 200 U/ml, an IgE response was observed in 11/26 cultures of PBMC from normal donors and in 12/15 cultures from allergic individuals. The same rIL-4-stimulated cells released significant amounts of IgE-binding factors (IgE-BF) in their culture supernatant. These IgE BF were shown for the first time to bind simultaneously to some mAb against Fc epsilon R II (mAbER) and to soluble IgE. The lack of correlation between the rIL 4-induced secretion of IgE-BF and IgE indicates that the production of IgE-BF or the expression of Fc epsilon R II is not the only factor involved in the induction of IgE synthesis by rIL-4. However, the observation that mAbER suppressed the rIL-4-induced IgE synthesis strongly suggests that either Fc epsilon R II or IgE-BF are necessary for an IgE response. Finally, the spontaneous in vitro synthesis of IgE by enriched B cell preparations isolated from atopic donors was suppressed in an isotype-specific manner by the same mAbER or by their F(ab')2 fragments. These observations suggest that the ongoing IgE synthesis by in vivo pre-activated B cells is also regulated by IgE-BF or Fc epsilon R II. It is concluded that Fc epsilon R II or IgE-BF play an essential role both in the induction of IgE synthesis by normal B cells and in the ongoing IgE synthesis by in vivo activated B cells from allergic donors. PMID- 2971722 TI - Characterization of the effector mechanism of help for antigen-presenting and bystander resting B cell growth mediated by Ia-restricted Th2 helper T cell lines. AB - The mechanism of help for resting B cell growth in MHC-restricted T-B collaboration was investigated using an in vitro polyclonal model for these T cell-B cell interactions. In the presence of rabbit anti-mouse Ig, small, size selected B cells elicit help from syngeneic Ia-restricted Th2 cell lines specific for F(ab')2 rabbit globulin. Both Ag-presenting and bystander B cells receive signals from stimulated Th cells that lead to B cell proliferation. The results suggest that the direct activation of resting Ag-presenting and bystander B cells by Th2 cells is mediated by a similar effector mechanism. Although proliferative responses by Ag-presenting B cells are of greater magnitude, help for both Ag presenting and bystander B cell populations is characterized by the lack of a requirement for membrane Ig cross-linking, by identical kinetics, and by the necessity for direct cell contact or close proximity with Th cells. B cell proliferation is not induced by exposure to the sequence of diffusable mediators released from a synchronized Ag-specific T-B interaction. The T cell-dependent proliferation by both B cell populations can be inhibited by excess mitomycin C treated syngeneic "cold target" B cells, demonstrating a requirement for a short range T cell-B cell interaction. mAb inhibition experiments fail to identify a role for class II, LFA-1, or CD4 membrane molecules in the delivery of help to bystander B cells. Antibody against H2d bystander class II molecules has no effect on bystander B cell proliferation at concentrations that completely block Ag presentation by H2d B cells to an H2d-restricted Th cell line. Antibodies against the cell adhesion molecule LFA-1 or the Th cell molecule CD4 do inhibit bystander B cell proliferation, but only to the extent that they block T cell activation and the induction of help. The inductive stimulus leading to resting B cell growth results from an early, short-range interaction with Th cells. B cell proliferation is supported by T cell soluble mediators as a consequence of this interaction, which is required for at least 8 hr after T cell recognition of Ag/Ia on the surface of Ag-presenting B cells. PMID- 2971723 TI - Identification of a T cell subset using a rabbit antibody raised against a T suppressor molecule. AB - Rabbit antiserum to a unique component of an Ag-binding Ts-factor was generated by repeated immunization with purified 30-kDa protein isolated from Fd11 Ts factor (11). This antiserum (anti-p30) was shown to recognize cell surface determinants expressed on the Ts hybridomas Fd11 and A10 but not the fusion partner BW5147. Furthermore, this antiserum was shown to bind to approximately 4% of thymocytes and 10% of nylon wool-purified splenic T cells from all strains of mice tested. Sorting nylon wool-purified T cells from DBA/2 mice for the CD4+ and CD8+ subsets revealed both populations contained cells that bound anti-p30. In addition, when CD4-8- thymocytes were examined for anti-p30 labeling, it was found that about 30% of this enriched population also expressed p30 molecules. In a functional study, anti-p30 was able to neutralize the suppressive effects of Fd11 on a specific assay for in vitro antibody synthesis against ferredoxin. PMID- 2971724 TI - Role of ornithine decarboxylase in the regulation of cell growth by IL-1 and tumor necrosis factor. AB - Ornithine decarboxylase (ODC) is a rate-limiting enzyme in polyamine synthesis, and polyamines are required for cell growth. As an approach to clarifying the mechanism of action IL-1, the effects of IL-1 on ODC activity were examined in various cell lines whose proliferation was either suppressed or enhanced by IL-1. The proliferation of all cell types used in these experiments was markedly suppressed by a specific ODC inhibitor, alpha-difluoromethyl ornithine (DFMO), substantiating the crucial role of ODC activity for cell proliferation. ODC activity also was considerably suppressed by IL-1 in those cells on which IL-1 exerts an antiproliferative effect, such as a human melanoma cell line (A375) and malignant human mammary cell lines (MCF-7 and T-47D). On the other hand, ODC activity was stimulated in cells that are stimulated to proliferate in response to IL-1, such as a mouse helper T cell line (D10.G4.1), a NK cell-like cell line (YT), and a human glioblastoma cell line (U373 MG). The effect of IL-1 on ODC activity preceded and directly correlated in a dose-dependent manner with its effect on DNA synthesis. Furthermore, putrescine, a product of the ODC reaction and a precursor of polyamines, was able to overcome most, but not all, the antiproliferative action of IL-1 in A375 melanoma cells, which were the most sensitive to suppression by IL-1. However, putrescine did not reverse the cytostatic effect of IL-1 on MCF-7 and T-47D cell lines. In contrast, putrescine, like IL-1, exhibited some co-mitogenic activity on D10.G4.1 cells. Because the biological activities of TNF and IL-1 show considerable overlap, the effect of TNF on ODC activity also was examined. TNF had an antiproliferative effect on A375 cells and stimulated the proliferation of U373 MG cells. The ODC activity in A375 cells was suppressed by TNF, and the ODC activity in U373 MG cells was stimulated by TNF. Putrescine also partially overcame the inhibitory effect of TNF. These results suggest that the regulation of ODC activity may be a key component in the antiproliferative and proliferative action of IL-1 and TNF in some tumor cell types. PMID- 2971725 TI - IgE receptor on human lymphocytes. IV. Further analysis of its structure and of the role of N-linked carbohydrates. AB - Previous studies have shown that the low affinity receptor for IgE (Fc epsilon R II) on human B lymphocytes was comprised of three components with apparent Mr of 45, 65 to 95, and 37 kDa. The present results indicate that the 37-kDa component is a soluble degradation fragment of the 45-kDa component and they also suggest that the 65- to 95-kDa component is probably made of aggregates of the above components that are formed after solubilization of the cells. The 45-kDa component appears to be a glycoprotein containing several sialic acid residues, O linked carbohydrates and one N-linked carbohydrate chain that is of the complex type. Partial digestion of the purified 65- to 95-, 45-, and 37-kDa molecules with alpha-chymotrypsin or protease V8 generates several fragments with identical mobility on SDS-PAGE. The 37 kDa is not N-glycosylated but like the IgE-binding factors present in the culture supernatant of Fc epsilon R-bearing cells, it contains sialic acid and O-linked carbohydrates. On incubation with protease inhibitors, the Mr of IgE-binding factors (BF) is shifted from 25-27 to 37 kDa, indicating that IgE-BF are derived from the proteolytic cleavage of the 37-kDa molecule, previously identified as a membrane component of Fc epsilon R. On incubation with N-glycosylation inhibitors, the production of IgE-BF is significantly increased indicating that N-glycosylation inhibits the degradation of Fc epsilon R into IgE-BF. Inasmuch as the effect of glycosylation inhibitors is not prevented by monensin, it is concluded that all the IgE-BF are derived from surface Fc epsilon R and not from their intracellular precursors. PMID- 2971726 TI - Anti-proliferative effect of IFN-gamma in immune regulation. II. IFN-gamma inhibits the proliferation of murine bone marrow cells stimulated with IL-3, IL 4, or granulocyte-macrophage colony-stimulating factor. AB - A biphasic dose response curve was observed when the bone marrow-derived cell line FDCP1, used as an indicator line for IL-3 bioassays, was exposed to supernatants from some activated T cell clones but not others. The active component which inhibited proliferation at the higher supernatant concentrations appeared to be IFN-gamma, based on the following observations. 1) Only those culture supernatants which contained IFN-gamma gave a biphasic dose response curve; 2) with these supernatants, an anti-IFN-gamma mAb augmented the proliferation of FDCP1 cells at the higher supernatant concentrations; and 3) rIFN-gamma profoundly inhibited the proliferation of FDCP1 cells stimulated with rIL-3 or rIL-4. rTNF-alpha inhibited FDCP1 proliferation only to a modest extent, yet the combination of rTNF-alpha + rIFN-gamma provided greater inhibition than each agent alone. The proliferation of a second bone marrow-derived cell line, DA1, was not inhibited by rIFN-gamma or rIFN-gamma + rTNF-alpha when stimulated with rIL-3 or recombinant granulocyte-macrophage colony-stimulating factor (rGM CSF). Fresh bone marrow cells also showed a suboptimal proliferative response when stimulated with T cell supernatants containing IFN-gamma, and this response was augmented considerably upon the addition of anti-IFN-gamma mAb. Bone marrow cell proliferation was observed upon exposure to rIL-3, rIL-4, or rGM-CSF, and these responses were inhibited by rIFN-gamma; rTNF-alpha also produced a synergistic effect with these cells. Bone marrow cell colony formation stimulated by rIL-3 or rGM-CSF also was inhibited by rIFN-gamma. Colony formation in bone marrow cell cultures was not observed in response to rIL-4. Collectively, these results suggest that Th1 cells, which in addition to IL-3 and GM-CSF also produce IFN-gamma, may regulate hemopoietic cell proliferation and colony formation differently from the way Th2 cells do, which do not produce IFN-gamma. PMID- 2971727 TI - In vivo restoration of T cell functions by human IL-1 beta or its 163-171 nonapeptide in immunodepressed mice. AB - The immunorestorative capacities of human (hu) IL-1 beta or its synthetic fragment 163-171 (VQGEESNDK) were assessed in vivo in mice immunodepressed by aging, sublethal irradiation, or both. Subcutaneous administration of hu rIL-1 beta into immunodepressed animals immediately after carrier (horse red blood cells, HRBC) priming could restore to normal levels Th cell activity. This was measured as the ability of spleen cells from HRBC-primed mice to induce a hapten specific antibody response in spleen cells from nonimmune mice in vitro stimulated with the hapten-carrier conjugate TNP-HRBC. In parallel, the ability of spleen cells from hu rIL-1 beta-treated immunodepressed animals to produce T cell growth factor activity upon in vitro mitogen stimulation was also increased significantly as compared to that of untreated mice and approached that of immunocompetent controls. The immunorestorative activity of hu rIL-1 beta on Th cell activity and T cell growth factor production could be mimicked by the synthetic nonapeptide 163-171 which, at the doses used, produced in most instances even greater effects than the whole protein. Although the optimal immunorestorative doses of the 163-171 peptide were several orders of magnitude higher than those of hu rIL-1 beta, the complete lack of IL-1-like inflammatory and toxic effects suggests that the synthetic hu IL-1 beta fragment may be successfully used as immunomodulating agent in the therapy of T cell immunodeficiencies. PMID- 2971728 TI - Structural polymorphism of the human platelet Fc gamma receptor. AB - A variable T lymphocyte proliferative response to murine IgG1 anti-T3 monoclonal antibodies, in which most North American Caucasians respond whereas a minority do not, is well established. This is most likely the result of a genetic polymorphism manifested by 1) the inability of the monocyte 40-kDa IgG FcR of some individuals to bind murine IgG1, and 2) a distinctive trimorphic pattern on IEF of the monocyte 40-kDa FcR, one form being seen in all individuals who do not respond and another form (or a combination of both forms) being seen in those who do respond. We have evaluated the IEF patterns of the platelet 40-kDa FcR and find that in every individual tested the pattern for platelet FcR correlates with that seen for the monocyte 40-kDa FcR pattern. Furthermore, the platelets of those individuals whose "nonresponder" monocyte 40-kDa FcR did not mediate a murine IgG1 anti-T3 response did not respond with an aggregation reaction to murine IgG1 immune complexes (opsonized E). In contrast, platelets from donors possessing "responder" monocytes displayed positive "aggregation" responses to E coated with murine IgG1 antibody. However, the platelet FcR structural polymorphism described earlier did not correlate with the donor-specific variability in capacity of platelets to respond functionally to aggregated human IgG described in an earlier paper. Rather, the variation in capacity of platelets from individual donors to respond functionally to aggregated human IgG was related to the quantitative expression of platelet FcR. These data indicate that the molecular mechanisms responsible for the platelet 40-kDa FcR structural polymorphism are quite different from the mechanisms governing the variation in quantitative expression of the receptor. PMID- 2971729 TI - Defective production of and response to IL-2 in acute human falciparum malaria. AB - Patients with acute Plasmodium falciparum malaria have defective cell-mediated immune responses to malaria-specific Ag (MA). This immunologic defect may partially explain the difficulty with which natural immunity to falciparum malaria develops and may have important implications for the efficacy of potential malaria vaccines in endemic areas. To investigate the basis of this immune defect, we have examined the capacity of PBMC from patients with acute falciparum malaria to produce IL-2 and to express I1-2R in response to Ag stimulation. The effect of exogenous IL-1 and IL-2 on lymphocyte proliferation was studied. Soluble IL-2R levels were measured in acute and convalescent sera. Our results showed that no detectable IL-2 was produced and no IL-2R were expressed by PBMC in response to MA during the acute infection. IL-2 production and IL-2R expression were also depressed when PBMC were exposed to streptococcal Ag. The specific immune defect was not reconstituted by the addition of graded doses of purified human IL-1 or IL-2 and could not be attributed to suppressor adherent cells. In contrast to the absence of IL-2 and cell-bound IL-2R, circulating soluble IL-2R was elevated in acute sera. These findings suggest that the lack of IL-2, through either a defect in its production or inhibition of its activity, may be the basis of the Ag-specific immune unresponsiveness in acute P. falciparum malaria. PMID- 2971730 TI - Complete exon-intron organization of the human leukocyte common antigen (CD45) gene. AB - Ten genomic DNA clones encoding the human leukocyte common Ag (LCA, CD45) gene were isolated by screening human genomic DNA libraries with LCA cDNA probes. One genomic DNA clone contains the promoter region and the first two exons, as determined by primer extension analyses and S1 nuclease protection studies as well as nucleotide sequence determination. The first exon does not encode a peptide, while the second exon contains the initiation ATG codon and encodes the signal peptide. The other nine genomic DNA clones, which are separated from the first genomic clone by an unknown distance, are connected and span a total of 73 kb. The nine connected genomic clones encode a total of 31 exons. The 33 exons encoded by these 10 genomic clones account for the entire cDNA sequences including the 5' and 3' untranslated sequences. Exon 3 and exons 7 through 15 encode the extracellular domain sequences that are common to all LCA isoforms. Differential usage of exons 4, 5, and 6, generates at least five distinct LCA isoforms. Exon 16 encodes the transmembrane peptide. The cytoplasmic region of the leukocyte common antigens is composed of two homologous domains. Exons 17 through 24 encode the first domain, and exons 25 through 32 encode the second domain. The comparison of these exons indicated that the homologous domains were generated by duplication of several exons. The most 3' exon (exon 33) encodes the carboxy terminus of the LCA molecules and includes the entire 3' untranslated sequence. PMID- 2971731 TI - Receptors for Fc epsilon and Fc gamma are linked on mouse chromosome 1. AB - Recently isolated cDNA clones for the high affinity Fc epsilon receptors on mast cells and basophils (Fc epsilon RI alpha) and Fc gamma receptors on macrophages and lymphocytes (Fc gamma 2b/gamma 1R) are homologous members of the Ig supergene family. Analysis of the segregation of restriction fragment length polymorphism in crosses of inbred mice now establish that the structural genes encoding both Fc epsilon RI alpha and Fc gamma 2b/gamma 1R are indeed discrete genes and are linked at the distal end of mouse chromosome 1. This finding raises the possibility that a family of Fc receptors could be found in a region that is known to contain immunologically important markers of lymphocyte surface Ag and autoimmune defects. PMID- 2971732 TI - Synergistic antitumor activity with IFN and monoclonal anti-idiotype for murine B cell lymphoma. Mechanism of action. AB - Combination therapy with syngeneic anti-idiotype antibody and human hybrid rIFN alpha A/D synergistically increase survival in C3H/HeN mice challenged with a lethal dose of tumor cells. C3H/HeJ mice, which have previously been described to be LPS hyporesponsive and have a defect in Fc gamma R function, did not respond to anti-idiotype therapy as well as C3H/HeN normal mice. This defect was completely corrected in animals treated simultaneously with IFN. Anti-idiotype mAb that was cleaved into F(ab')2 fragments no longer had any antitumor activity alone and could not be enhanced by IFN therapy. These results suggest that antibody is functioning through Fc gamma R-bearing effector cells that are enhanced by IFN therapy. Synergy between IFN and anti-idiotype mAb was maintained in nude mice lacking classical T cells but was reduced in C3H beige mice lacking classical NK/killer cells. IFN did not increase idiotype expression on the tumor cells but did increase H-2 expression. Although we have previously shown that rIFN-alpha A/D can directly kill 38C13 in vitro, an IFN-resistant subclone derived from 38C13, SIR-1, was equally or more responsive to human rIFN-alpha A/D in vivo and had a synergistic antitumor response to combination IFN and anti idiotype therapy, indicating that IFN acts primarily through host mediated effects rather than direct effects. PMID- 2971733 TI - Plasticity of T cell function. Cloned EL-4 lymphoma cells may help or suppress a primary antibody response depending on their own concentration and the assay system. AB - Cloned EL-4 lymphoma T cells were tested in limiting dilution experiments for their capacity to suppress or to help the primary humoral immune response of spleen cells (or T cell-depleted spleen cells) to the Ag SRBC and 4-hydroxy-3 iodo-5-nitro-phenyl-keyhole limpet hemocyanin. EL-4 clones are able to suppress up to 80% of the total IgM responses in both systems, as well as to help. Suppression and help fluctuate between high and low levels with the numbers of EL 4 cells placed into tissue cultures. In Poisson plots, this is reflected as a "typical curve": usually one or two frequencies are estimated (e.g., integral of 1/4 and approximately 1/1000 for suppression and approximately 1/6 and approximately 1/200 for help), which appear regulated with increasing numbers of cells seeded. Control experiments showed that EL-4 cells need to be alive to exert the effects. EL-4 cells do not serve as additional antigen, do not induce an isotype switch and are not cytotoxic. Help and suppression are not restricted by the MHC. Help requires the presence of a small number of normal T cells in the assay system, indicating that EL-4 cells do not replace specific helper T cells. When a number of control cell lines were analyzed under identical circumstances, similar effects were observed with most long term T cell lines or clones expressing a T cell receptor, whereas cells of non-T lineages and T cells not expressing a T cell receptor did not show the phenomena. The results suggest a functional plasticity of T cells, dependent on cell numbers and the assay system used, and expressed via T cell communication. PMID- 2971734 TI - Inhibition of anti-IgM and growth factor-induced proliferation of guinea pig B cells by one of two distinct Fc gamma receptors on the cells. AB - Guinea pig B cells were found to proliferate when co-stimulated with F(ab')2 of rabbit anti-guinea pig IgM and human 12-kDa B cell growth factor (BCGF), though the proliferation did not occur with the replacement of the F(ab')2 by its parent IgG antibody. In addition, the intact antibody inhibited the proliferation induced by F(ab')2 of anti-IgM and BCGF. Because both two distinct types of FcR for IgG on the B cells, one specific for IgG2 (Fc gamma 2R) and the other for both IgG2 and IgG1 (Fc gamma 1/gamma 2R), can bind rabbit IgG, we determined whether they participate in the inhibition of the B cell proliferation by intact anti-guinea pig IgM antibody. Blocking Fc gamma 1/gamma 2R by F(ab')2 of anti-Fc gamma 1/gamma 2R mAb significantly reversed the inhibitory effect of intact anti IgM antibody. F(ab')2 of anti-Fc gamma 2R mAb, however, was not effective. Furthermore, guinea pig IgG1 and IgG2 anti-rabbit IgG antibodies suppressed similarly the B cell proliferation induced by F(ab')2 of rabbit anti-IgM and BCGF. These results show that between these two types of Fc gamma R on B cells, Fc gamma 1/gamma 2R alone is involved in the regulation of anti-IgM and BCGF induced B cell proliferation, and inhibits the response when cross-linked to the surface IgM. PMID- 2971736 TI - Production of human bone marrow-derived suppressor factor. Effect on antibody synthesis and lectin-activated cell proliferation. AB - Natural suppressor cells resident in normal human bone marrow (BM) exert potent suppressor activity on in vitro antibody responses and other immune functions. A suppressor-enriched population of BM cells can constitutively produce a soluble mediator with similar suppressor activity and kinetics as the suppressor cells. This novel BM-derived suppressor factor (BDSF) suppresses human in vitro primary antibody responses as well as lectin-activated proliferative responses. The mediator (BDSF) has a Mr of less than 1.5 kDa, contains a lipid component, and is insensitive to indomethacin treatment. The BM cells producing the factor bear the HNK-1 surface marker but not T, B, or macrophage markers. The ability of BDSF to suppress Ag-dependent IgM responses during the inductive phase makes it an ideal molecule with the potential to regulate early immune and hemopoietic events within the BM compartment. PMID- 2971735 TI - Alloantigen presentation by B cells. Requirement for IL-1 and IL-6. AB - We have evaluated the requirements for non-activated allogeneic B cells to present alloantigen in primary MLC. We found that the generation of an MLR was dependent on the addition of exogenous IL-1 and IL-6 to the cultures; neither lymphokine by itself was capable of significantly stimulating the response. Our results suggest that the concentration of these lymphokines present in a standard MLR culture may serve to regulate the magnitude of the proliferative response. The B cells that co-stimulated with IL-1 and IL-6 were found to be contained almost exclusively within the large, low density population. Presumably, large low density B cells have attained a state in which they are particularly effective at presentation of alloantigen. Together our findings indicate that three components are required for Ag presentation in primary MLC: class II MHC positive stimulator cells that have attained the appropriate state of differentiation and the presence of both IL-1 and IL-6. PMID- 2971737 TI - Leukocyte inhibitory factor stimulates neutrophil-endothelial cell adhesion. AB - We investigated the ability of the human lymphokine leukocyte inhibitory factor (LIF) to modulate neutrophil-endothelial cell (EC) adherence. EC were cultured from collagenase-treated human umbilical cord veins and grown in complete medium supplemented with EC growth factor. Adherence was measured as the percent of 51Cr labeled neutrophils remaining adherent to the EC after gentle lavage. Polymorphonuclear neutrophils (PMN) were pretreated with LIF (0.5 to 8 U/ml), extensively washed, and allowed to interact with the EC monolayers. LIF was demonstrated to induce an increase in the capacity of PMN to bind EC in a dose dependent fashion (from 30.9 +/- 2.1% adherence with control-treated PMN to 68.6 +/- 3.0% at 4 U LIF; p less than 0.001). In subsequent experiments we demonstrated that 10 min was a sufficient preincubation time for LIF to modulate the capacity of the PMN to adhere to EC. LIF has previously been observed to up regulate expression of C receptor type 3 on PMN, a receptor which has been shown to be involved in PMN-EC binding. Exposure of PMN to anti-C receptor type 3 antibody before their incubation with LIF abrogated its effect as did inactivation of LIF by an esterase inhibitor. We also investigated the ability of LIF to stimulate EC to bind untreated PMN. EC were pretreated with LIF (0.25 to 4 U/ml), extensively washed, and adherence measured as before. LIF was shown to induce a dose-dependent increase in the capacity of the EC to bind PMN (from 28.8 +/- 3.1% for untreated EC to 91.1 +/- 4.0% at 4 U LIF; p less than 0.001). Modulation of EC function required a minimum of 30 min and was inhibited in the presence of cycloheximide or actinomycin D. Neither anti-TNF-alpha or -beta antibodies nor polymixin B abrogated the augmentation by LIF. However, anti-IL-1 antibody partially inhibited the stimulation of EC adhesiveness by LIF, suggesting the possible involvement of this cytokine. These studies provide further evidence that LIF may mediate an important pro-inflammatory role in vivo. PMID- 2971738 TI - Enrichment of murine splenic natural killer (NK) cells by the sequential elimination of non-NK cells. AB - A simple and reliable three-step procedure to enrich for murine endogenous splenic NK cells is described. The method is based on the sequential elimination of non-NK cell subsets by standard and inexpensive techniques executed in a specific order. First, macrophages and other adherent cells are eliminated by incubation on plastic surface. Secondly, the T cells are excluded from the multicellular aggregates formed by agglutination of the remaining cells with wheat germ lectin. Thirdly, after dissociation of the aggregates with N-acetyl-D glucosamine and osmotic lysis of erythrocytes, NK cells are separated from other nucleated cells by nylon wool filtration. C57BL/6 spleen cells were used to establish the enrichment procedure. Usually their NK cell activity is intermediate but occasionally either low or high NK cell activity was observed in input cell suspensions. The NK cell activity recovery and the degree of enrichment varied inversely with the initial NK cell activity level of the input cell suspension. When initial NK cell activity was intermediate, it was enriched 10-30-fold. Experiments were done to establish if suppressor cells, and nylon wool-adherent, naturally activated NK cells, putatively present in input cells, could have been responsible for the abnormal initial NK cell activity detected in some C57BL/6 spleen cell suspensions and for the variations in the degree of enrichment achieved by the method here described. Either no or negligeable suppressor cell activity was noted in the cell fractions normally discarded at each step of the procedure. On the other hand, nylon wool-adherent NK cells were eliminated during the fractionation of spleen cells with higher than average initial NK cell activity and would account for the lower NK cell enrichment obtained in these conditions. PMID- 2971739 TI - Photometric assays for FcRI-dependent binding, phagocytosis, and antibody dependent cellular cytotoxicity mediated by monomeric IgG gamma 2a in murine peritoneal macrophages. AB - Mouse peritoneal macrophages possess distinct Fc receptors (FcR) for binding the various murine IgG isotypes. FcRI binds monomeric IgG gamma 2a, but not monomeric IgG gamma 2b or IgG gamma 1 with high affinity at 4 degrees C and is sensitive to trypsin degradation. We have assessed the functional consequences of the cytophilic binding at 4 degrees C of monomeric IgG gamma 2a to FcRI of mouse peritoneal macrophages using newly developed photometric microassays for quantification of binding, phagocytosis, and antibody dependent cellular cytotoxicity (ADCC) of post-opsonized sheep red blood cell (SRBC) targets. Dose dependent binding specificity of monomeric IgG gamma 2a, but not IgG gamma 2b or IgG gamma 1 to FcRI of oil-elicited mouse peritoneal macrophages at 4 degrees C for 2 h was confirmed to display typical saturation kinetics both by the photometric assay and by a cellular enzyme-linked immunosorbent assay (CELISA). Binding of monomeric IgG gamma 2a to macrophage FcRI promoted highly efficient phagocytosis of opsonized SRBC in that most cells that were bound were also rapidly internalized by the phagocytic process during a 1 h incubation at 37 degrees C. Upregulation of FcRI-dependent binding and phagocytosis occurred during 24-48 h in vitro culture of macrophages as shown both by the photometric assays and CELISA. Trypsin treatment of macrophages abrogated FcRI-dependent binding and phagocytosis by monomeric IgG gamma 2a, but had little effect on FcRII-dependent functions. Cytophilic binding of monomeric IgG gamma 2a to FcRI failed to trigger ADCC activation. Thus functional characterization of macrophage FcRI-dependent effector functions confirmed the fidelity of binding specificity of monomeric IgG gamma 2a to a trypsin degradable receptor which mediates highly efficient phagocytosis but fails to initiate the signal for ADCC activation. It appears that passively bound immune monomeric IgG gamma 2a could provide an efficient mechanism by macrophages in vivo for FcRI-dependent immune clearance of soluble or particulate cellular antigens without elicitation of potentially harmful cytolytic factors associated with ADCC activation. PMID- 2971740 TI - Importance of CD8+ T helper cell function in AIDS. PMID- 2971742 TI - Self-nonself discrimination due to immunological nonlinearities: the analysis of a series of models by numerical methods. PMID- 2971743 TI - Medico-economic implications of industrial hand injuries in India. AB - 625 five consecutive cases of industrial hand injuries attending the Employee's State Insurance Hospital, Jaipur, have been studied from 1983 to October 1986. The incidence of injuries was 36 per 10,000 workers per year. 47% were due to entrapment of the hand in active machines, 25% occurred during lifting and transportation of heavy objects and 12% while handling tools. The injuries resulted in residual deficit in 55% of cases and were serious enough to require absence from work of more than four weeks in 48% of cases. On an average 35 days were lost per injured worker. The average economic loss per injured worker was Rs. 6900 (approximately pounds 275) for workers in the wage-range of Rs. 5400 to 19,200 (pounds 216 to pounds 768) per annum. PMID- 2971741 TI - Angioplasty for stable versus unstable angina pectoris: are unstable patients more likely to get restenosis? A quantitative angiographic study in 339 consecutive patients. AB - Current evidence with regard to the possible association between clinical expression of coronary disease prior to the time of angioplasty, and the subsequent risk of restenosis following successful dilatation, remains inconclusive. To prospectively compare the incidence of restenosis in stable versus unstable angina pectoris patients, follow-up angiography was performed in 85 percent of patients from a consecutive series with a successful PTCA, irrespective of presence or absence of recurrent ischemic symptoms. Furthermore, changes in lesion severity were assessed quantitatively by an automated edge detection technique rather than visual analysis. Employing such a study design and follow-up protocol, it was found that the incidence of restenosis in patients with stable coronary artery disease was similar to that of patients with unstable rest angina, irrespective of the type of angiographic definition used. PMID- 2971744 TI - Ipratropium bromide in the treatment of the 'rhinorrhoea syndrome'. AB - Conventional treatments for non-allergic perennial rhinitis have proven somewhat unsuccessful in the control of rhinorrhoea when it is the predominant symptom. Hence, a double-blind cross-over trial of Ipratropium, a parasympatholytic, and placebo were carried out over a 12-week period. There was a significant reduction in rhinorrhoea during active treatment, with the most noticeable effect being in the moderate-to-severe rhinorrhoea group. No significant effect was noted on nasal obstruction or sneezing and no serious side-effects were seen. A carry-on effect was noted when active treatment was used initially. However, the group given active treatment following placebo had a better result overall. A strong placebo effect was also noted in both groups, in keeping with a trial of this order. PMID- 2971745 TI - [Contribution of femoral hemodynamics (intra-arterial velocity, pressure) in indications for vascular reconstruction in ischemia of the lower limbs. Preliminary study]. AB - The choice of the best operative procedure in vascular reconstructions for ischemic lower limbs, is mainly guided by clinical experience and by arteriography. Doppler wave analysis and femoral artery pressure measurements are new diagnostic tools. In those difficult cases of multiple-level-disease, the intra-operative measurement of the femoral artery pressure proved to be an important step for the operative decision. PMID- 2971747 TI - An analytical survey of structural aberrations observed in static radiographic examinations among acute low back cases. PMID- 2971748 TI - Exercise and smoking habits in patients with and without low back pain and leg pain. PMID- 2971746 TI - [Percutaneous angioscopy of the iliac and femoral arteries]. AB - Very few papers about peripheral angioscopy are reported in literature. Percutaneous angioscopy (P.T.A.) of 25 peripheral arteries (21 iliac and 4 femoral arteries) have been performed by the authors without surgery and without anesthesia. Three observations are selected. The first one demonstrates an eccentric stenosis altering its diameter during pulsations; an irregular ulcerated atheroma is observed. The second case shows the signs of a centric atheroma with an intimal fragment. The last one is an intimal dissection due to PTA. The prospective aspects of this new technique are discussed. PMID- 2971749 TI - Diagnostic importance of fibrillatory wave amplitude: a clue to echocardiographic left atrial size and etiology of atrial fibrillation. AB - The electrocardiographic fibrillatory wave amplitude in 148 patients with atrial fibrillation was measured. To assess its diagnostic importance in relation to the left atrial size and underlying etiology, M-mode echocardiography was performed on 89 patients. The patients are classified according to the type of atrial fibrillation (paroxysmal or chronic), the f wave amplitude (coarse or fine), and the underlying etiology (rheumatic or nonrheumatic). The average fibrillatory f wave amplitude and echocardiographic left atrial size in chronic and rheumatic atrial fibrillation are significantly greater than in paroxysmal and chronic nonrheumatic patients. Furthermore, the fibrillatory f wave amplitude correlates strongly with the echocardiographic left atrial size. PMID- 2971750 TI - Secretion of beta-endorphin into the maternal circulation by uteroplacental tissues in response to hypoglycaemic stress. AB - The placenta has been shown to contain ACTH and beta-endorphin but the roles of these peptides are unknown. To investigate whether they are released into the maternal circulation from the placenta in response to physiological stimuli the effects of hypoglycaemic stress were investigated. Plasma samples were collected from the femoral artery (FA) and uterovarian (UV) vein of nine pregnant sheep before and during hypoglycaemia induced by intravenous insulin (100U). Plasma concentrations of ovine beta-endorphin (o beta-EP) were measured by radioimmunoassay. Concentrations of o beta-EP rose in both vessels by 60 min after insulin. The peak concentrations of o beta-EP (pmol/l) were 122 +/- 29 (mean +/- SEM, n = 8) in the UV and 96 +/- 24 (n = 9) fmol/ml in the FA 60 min after insulin injection. There was no difference between the concentrations of o beta-EP in the vessels before insulin injection but at 60 and 120 min after insulin the concentrations of o beta-EP were significantly higher in the UV than FA (P less than 0.02, analysis of variance). This indicates that the pregnant uterus or placenta can respond to hypoglycaemia by secreting beta-EP into the maternal circulation. It is therefore possible that placental pro opiomelanocortin (POMC) peptides may have a role in maternal endocrinology and metabolism. PMID- 2971751 TI - Surface expression of only gamma delta and/or alpha beta T cell receptor heterodimers by cells with four (alpha, beta, gamma, delta) functional receptor chains. AB - Surface expression of TCR dimers by cells synthesizing three or four distinct types of receptor chains was analyzed. Cells containing intact gamma, alpha, and beta chains had only gamma delta dimers on the cell surface. In human PEER cells, addition of a functional alpha chain led to the loss of gamma delta dimer expression and expression of only alpha beta dimers. This result was not due to transcriptional down-regulation of the gamma or delta loci. In murine cells expressing all four chains, both gamma delta and alpha beta dimers could be demonstrated on a single cell. No other chain combinations (alpha gamma, alpha delta, beta gamma, or beta delta) were detected. Thus, there is stringent control of assembly and/or transport of TCR heterodimers, such that functional receptors consist only of alpha beta and gamma delta pairs, and no additional repertoire diversity is generated by cross pairing. PMID- 2971752 TI - Predominant variable region gene usage by gamma/delta T cell receptor-bearing cells in the adult thymus. AB - Previous studies have indicated that the diversity of gamma genes expressed by gamma/delta-bearing murine T cells is limited, but comparable information concerning the expressed diversity of delta genes is lacking. In this study, we have investigated the rearrangement and expression of delta and gamma genes in T cell hybridomas that express gamma/delta T cell receptors. Three productive delta chain cDNA clones were isolated (delta 7.3, delta 7.1, and delta 2.3) that encode new variable region sequences. Two of the delta cDNAs differ significantly from those observed in the V alpha repertoire. In addition, one cDNA expressed a new J delta region (J delta 2), which was localized between J delta 1 and C delta genes. Using these and other delta gene probes and gamma gene probes, we found that five independent hybridomas expressed four different V delta s and three different V gamma s. However, analysis of an enriched population of gamma/delta expressing cells from the adult thymus suggests that only a few V delta genes and one V gamma gene are used by the majority of the cells. These results suggest that important components of receptor chain that contribute to specificity (i.e., the germline V gene sequences) are relatively nondiverse in the thymic gamma/delta population. PMID- 2971753 TI - T cell receptor gene usage in the response to lambda repressor cI protein. An apparent bias in the usage of a V alpha gene element. AB - The T cell response to the lambda repressor cI protein is directed to the same region of the protein (residues 12-26) in both BALB/c and A/J mice. A panel of T cell hybridomas specific for P12-26 in the context of either I-Ek or I-Ad have been isolated To further understand the molecular interaction between the TCR and the Ia-P12-26 complex, the primary structures of the TCR of five T cell hybridomas have been determined. Southern and Northern analyses indicate that two members of the V alpha 3 gene family are used by 13 out of 14 I-Ek-restricted T cells. Four different V beta genes are used by these T cell hybridomas, while the majority (8 out of 13) express V beta 1 in combination with the J beta 2.1 element. No clear correlation can be seen in this system between gene usage and MHC restriction. In addition, the fine specificity of I-Ek-restricted T cells to a single amino acid substitution [Phe22/His22]P12-26 is not attributed to the usage of particular V alpha and V beta elements. The V alpha 3 family gene is also used by a few I-Ad-restricted T cells. Interestingly, these I-Ad T cells share a reactivity pattern more similar to that of I-Ek-restricted T cells than other I-Ad-restricted T cells. The nonrandom selection V alpha 3 is also demonstrated by the fact that V alpha 3 is used by P12-26-specific, but not by cytochrome c- or staphylococcal nucleus-specific, I-Ek-restricted T cells. This suggests that although antigen specificity may not be accounted for by either chain of the TCR, the members of V alpha 3 genes may be selected by the antigen (P12-26). PMID- 2971754 TI - Thymic selection of H-2-incompatible bone marrow cells in SCID mice. Differences in T help for induction of B cell IgG responses versus cytotoxic T cells. AB - Mice with congenital severe combined immunodeficiency disease (SCID) failed to mount either a T cell-independent IgM or T cell-dependent IgG anti-vesicular stomatitis virus (VSV) Indiana (IND) response. They did not generate cytotoxic T cells against lymphocytic choriomeningitis virus (LCMV) or vaccinia virus, but exhibited NK cell-like activities. When SCID mice were given bone marrow from syngeneic BALB/c (H-2d) nu/nu mice, all immune responses were expressed at control levels. If SCID mice were reconstituted with allogeneic H-2b C57BL/6 nu/nu bone marrow, the following primary anti-viral immune responses were measured. T-independent IgM anti-VSV-IND were normal, but T-dependent IgG anti VSV-IND responses were absent. Cytotoxic T cell responses to LCMV and vaccinia virus were within normal ranges, were donor cell mediated, and were specific exclusively for the recipient SCID H-2d type. Since antigen presentation by spleen cells was functional in these chimaeras, the presented results indicate that (a) thymic selection of T cell restriction is strict; and (b) the type of T help necessary for B cells depends upon H-2-restricted contact between T and B cells, whereas, such contact-dependent help is not mandatory for the induction of virus-specific cytotoxic T cells. PMID- 2971755 TI - Heterogeneity of lymphocyte calcium metabolism is caused by T cell-specific calcium-sensitive potassium channel and sensitivity of the calcium ATPase pump to membrane potential. AB - Calcium management differs in T and B lymphocytes. [Ca2+]i elevation in response to calcium ionophores is up to 10 times greater in T cells than B cells. There is no difference between them in ionophore uptake. T cells, but not B cells, possess a calcium-sensitive potassium channel which produces membrane hyperpolarization at [Ca2+]i above 200 nM. This alters T cell density providing a rapid and easy method of cell separation. In contrast, B cells depolarize when [Ca2+]i is increased. Isolated B cell membrane vesicle ATP-dependent calcium pump activity is higher than T cell vesicles. Membrane depolarization reduces the [Ca2+]i response to ionomycin, most dramatically in T cells because they are hyperpolarized by increased [Ca2+]i. The most likely basis of this behavior is an effect of membrane potential on lymphocyte membrane calcium pump activity. This mechanism provides an explanation of the inhibitory effect of membrane depolarization on T lymphocyte responses. PMID- 2971756 TI - Common acute lymphocytic leukemia antigen is identical to neutral endopeptidase. AB - We purified CALLA from human kidney and isolated a cDNA clone reactive with two oligonucleotide probes corresponding to two distinct peptides. The amino acid sequence translated from the CALLA cDNA revealed 100% identity with that of human neutral endopeptidase (NEP, enkephalinase). The distribution of CALLA antigen and NEP in normal tissues are similar. PMID- 2971757 TI - Identification of an alternative polyadenylation site in the human C3b/C4b receptor (complement receptor type 1) transcriptional unit and prediction of a secreted form of complement receptor type 1. AB - The human C3b/C4b receptor or complement receptor type one (CR1) is an approximately 200-kD single chain membrane glycoprotein of human peripheral blood cells that mediates the binding, processing, and transport of C3b-bearing immune complexes and regulates the activity of the complement cascade. Analysis of partial cDNA clones has shown that the COOH terminus is composed predominantly of three tandemly repeated regions of 450 amino acids each (15). In this report, we present a cDNA sequence that encodes the NH2 terminus of CR1. It appears to have been derived from an alternatively processed transcript, caused by polyadenylation occurring at a site within an intron in the CR1 transcriptional unit. The resulting truncated messenger carries an open reading frame that would produce a short, secreted CR1 form. We present genomic sequences and Northern blots which support this hypothesis and we propose that the NH2-terminal end of CR1 is a likely location for active sites. In addition, we report evidence for a CR1-like sequence in the human genome and we present a model for the organization of CR1. PMID- 2971758 TI - Bacterial cell wall-induced immunosuppression. Role of transforming growth factor beta. AB - Group A streptococcal cell wall (SCW)-injected rats exhibit a profound immunosuppression that persists for months after the initial intraperitoneal injection of SCW. The goal of this study was to determine the mechanisms for the suppressed T lymphocyte proliferative responses in this experimental model of chronic inflammation. When spleen cell preparations were depleted of adherent cells, restoration of T cell proliferative responses to Con A and PHA occurred, implicating adherent macrophages in the regulation of immunosuppression. Furthermore, macrophages from SCW-treated animals, when cocultured with normal spleen cells in the presence of Con A or PHA, effectively inhibited the proliferative response. Supernatants from suppressed spleen cell cultures were found to inhibit normal T cell mitogenesis. Taken together, these results implicated a soluble macrophage-derived suppressor factor in the down regulation of T cell proliferation after exposure to SCW in vivo. Subsequent in vitro studies to identify this suppressor molecule(s) revealed the activity to be indistinguishable from the polypeptide transforming growth factor beta (TGF beta). Furthermore, TGF-beta was identified by immunolocalization within the spleens of SCW-injected animals. The cells within the spleen that stained positively for TGF-beta were phagocytic cells that had ingested, and were presumably activated by, the SCW. These studies document that TGF-beta, previously shown to be a potent immunosuppressive agent in vitro, also effectively inhibits immune function in chronic inflammatory lesions in vivo. PMID- 2971759 TI - Properties of isolated red pulp macrophages from mouse spleen. AB - Dense monolayers of large, adherent macrophages were prepared from the red pulp of mouse spleen. These sinus-lining phagocytes resembled liver Kupffer cells in morphology, as well as expression of F4/80 and class II MHC antigens and receptors for IgG. C3-coated red cells attached at low levels to spleen macrophages, but attachment and endocytosis were enhanced on fibronectin-coated surfaces. The ionophore A23187 induced spleen macrophages to synthesize prostaglandin E2, but like Kupffer cells, spleen macrophages did not synthesize leukotrienes and made relatively small amounts of HETE and 12 hydroxyheptadecanoic acid. Resident spleen macrophages did not produce H2O2, but splenic inflammatory cells, induced by infection of animals with Listeria monocytogenes, actively released H2O2. We conclude that the functional properties of resident, sinusoidal-lining macrophages in liver and spleen are similar to one another but distinct from other pools of phagocytes. PMID- 2971760 TI - From conditioning to category learning: an adaptive network model. AB - We used adaptive network theory to extend the Rescorla-Wagner (1972) least mean squares (LMS) model of associative learning to phenomena of human learning and judgment. In three experiments subjects learned to categorize hypothetical patients with particular symptom patterns as having certain diseases. When one disease is far more likely than another, the model predicts that subjects will substantially overestimate the diagnosticity of the more valid symptom for the rare disease. The results of Experiments 1 and 2 provide clear support for this prediction in contradistinction to predictions from probability matching, exemplar retrieval, or simple prototype learning models. Experiment 3 contrasted the adaptive network model with one predicting pattern-probability matching when patients always had four symptoms (chosen from four opponent pairs) rather than the presence or absence of each of four symptoms, as in Experiment 1. The results again support the Rescorla-Wagner LMS learning rule as embedded within an adaptive network model. PMID- 2971762 TI - Strategy choice procedures and the development of multiplication skill. AB - Many intelligent strategy choices may be accomplished through relatively low level cognitive processes. This article describes a detailed model of how such "mindless" processes might lead to intelligent choices of strategies in one common situation: that in which people need to choose between stating a retrieved answer and using a backup strategy. Several experiments testing the model's applicability to children's single-digit multiplication are reported. These include tests of predictions about when different strategies are used and how early experience shapes later performance. Then, the sufficiency of the model to generate both performance at any one time and changes in performance over time is tested through the medium of a running computer simulation of children's multiplication. The simulation acquires a considerable amount of multiplication knowledge, and its learning and performance parallel those of children in a number of ways. Finally, several implications of the model for understanding cognitive self-regulation and cognitive development are discussed. PMID- 2971761 TI - Electrophysiological evidence for a shared representational medium for visual images and visual percepts. AB - Does mental imagery involve the activation of representations in the visual system? Systematic effects of imagery on visual signal detection performance have been used to argue that imagery and the perceptual processing of stimuli interact at some common locus of activity (Farah, 1985). However, such a result is neutral with respect to the question of whether the interaction occurs during modality specific visual processing of the stimulus. If imagery affects stimulus processing at early, modality-specific stages of stimulus representation, this implies that the shared stimulus representations are visual, whereas if imagery affects stimulus processing only at later, amodal stages of stimulus representation, this implies that imagery involves more abstract, postvisual stimulus representations. To distinguish between these two possibilities, we repeated the earlier imagery-perception interaction experiment while recording event-related potentials (ERPs) to stimuli from 16 scalp electrodes. By observing the time course and scalp distribution of the effect of imagery on the ERP to stimuli, we can put constraints on the locus of the shared representations for imagery and perception. An effect of imagery was seen within 200 ms following stimulus presentation, at the latency of the first negative component of the visual ERP, localized at the occipital and posterior temporal regions of the scalp, that is, directly over visual cortex. This finding provides support for the claim that mental images interact with percepts in the visual system proper and hence that mental images are themselves visual representations. PMID- 2971763 TI - Visual masking and visual integration across saccadic eye movements. AB - The visual world appears unified, stable, and continuous despite rapid changes in eye position. How this is accomplished has puzzled psychologists for over a century. One possibility is that visual information from successive eye fixations is fused in memory according to environmental or spatiotopic coordinates. Evidence supporting this hypothesis was provided by Davidson, Fox, and Dick (1973). They presented a letter array in one fixation and a mask at one letter position in a subsequent fixation and found that the mask inhibited report of the letter that shared its retinal coordinates but appeared to occupy the same position as the letter that shared its spatial coordinates. This suggests the existence of a retinotopic visual persistence at which transsaccadic masking occurs and a spatiotopic visual persistence at which transsaccadic integration, or fusion, occurs. Using a similar procedure, we found retinotopic masking and retinotopic integration: The mask interfered with the letter that shared its retinal coordinates, but also appeared to cover that letter. In another experiment, instead of a mask we presented a bar marker over one letter position, and subjects reported the letter that appeared underneath the bar; subjects usually reported the letter with the same retinal coordinates as the bar, again suggesting retinotopic rather than spatiotopic integration across saccades. In Experiment 3 a bar marker was again presented over one letter position, but in addition a visual landmark was presented after the saccade so that subjects could localize the bar's spatial position; subjects still reported that the letter that shared the bar's retinal coordinates appeared to be under it, but they were also able to accurately specify the bar's spatial position. This ability could have been based on retinal information (the visual landmark) present in the second fixation only, however, rather than spatiotopic visual persistence. Because such a visual landmark was present in the Davidson et al. (1973) experiments, we conclude that their findings can be explained solely in retinotopic terms and provide no convincing evidence for spatiotopic visual persistence. But the exposure parameters that Davidson et al. (1973) and we used were biased in favor of retinotopic, rather than spatiotopic, coding: The stimuli were presented very briefly just before a saccadic eye movement, and subjects are poor at spatially localizing stimuli under these conditions. Thus, in Experiment 4 we presented the letter array about 200 ms before the saccade; then, subjects reported that the letter with the same spatial coordinates as the bar appeared under it.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2971765 TI - Examination of perceptual reorganization for nonnative speech contrasts: Zulu click discrimination by English-speaking adults and infants. AB - The language environment modifies the speech perception abilities found in early development. In particular, adults have difficulty perceiving many nonnative contrasts that young infants discriminate. The underlying perceptual reorganization apparently occurs by 10-12 months. According to one view, it depends on experiential effects on psychoacoustic mechanisms. Alternatively, phonological development has been held responsible, with perception influenced by whether the nonnative sounds occur allophonically in the native language. We hypothesized that a phonemic process appears around 10-12 months that assimilates speech sounds to native categories whenever possible; otherwise, they are perceived in auditory or phonetic (articulatory) terms. We tested this with English-speaking listeners by using Zulu click contrasts. Adults discriminated the click contrasts; performance on the most difficult (80% correct) was not diminished even when the most obvious acoustic difference was eliminated. Infants showed good discrimination of the acoustically modified contrast even by 12-14 months. Together with earlier reports of developmental change in perception of nonnative contrasts, these findings support a phonological explanation of language-specific reorganization in speech perception. PMID- 2971764 TI - Pre- and poststimulus activation of response channels: a psychophysiological analysis. AB - To examine mechanisms of response activation, we asked subjects to respond differentially to the central letter of one of four arrays--HHHHH, SSHSS, SSSSS, and HHSHH--and measured event-related brain potentials (ERPs) and electromyographic activity (EMG). For very fast responses, accuracy was at chance level for all arrays, suggesting that subjects were guessing. For intermediate latency responses, accuracy was above chance if the noise was compatible with the targets and below chance if it was incompatible, suggesting that these responses were based on partial stimulus analysis. For slow responses, accuracy was above chance for all arrays, suggesting that these responses were based on complete stimulus analysis. The occurrence and accuracy of fast responses could be predicted by examining motor potentials preceding the presentation of the array. Measures of the motor potentials in the period following the presentation of the array suggested that partial analysis of stimulus information could activate responses and that the level of response activation at the time of the EMG response was constant for trials with different response latencies. The data are discussed in terms of a response channel conception. PMID- 2971766 TI - Context effects in two-month-old infants' perception of labiodental/interdental fricative contrasts. AB - We investigated 2-month-old infants' perception of a subset of highly confusable English fricatives. In Experiment 1, infants discriminated modified natural tokens of the voiceless fricative pair [fa]/[oa] but only when the syllables included their frication noises. They also discriminated the voiced pair [va]/[oa] both with and without fricative noises. These results parallel those found with adults by Carden, Levitt, Jusczyk, and Walley (1981). In Experiment 2 [f] and [o] noises were appended to [a], and the same [f] noise was appended to the previously indiscriminable fricationless versions of [fa] and [oa]. Infants discriminated both pairs of stimuli, indicating that (a) the frication is a sufficient cue for [fa]/[oa] discrimination and that (b) it provides a context for discriminating the [f] and [o] formant transitions. We conclude that infants' perception of labiodental/interdental fricative contrasts show evidence of context effects similar to those observed with adults. PMID- 2971767 TI - Effects of speaking rate and lexical status on phonetic perception. AB - Among the contextual factors known to play a role in segmental perception are the rate at which the speech was produced and the lexical status of the item, that is, whether it is a meaningful word of the language. In a series of experiments on the word-initial /b/-/p/ voicing distinction, we investigated the conditions under which these factors operate during speech processing. The results indicated that under instructions of speeded responding, listeners could, on some trials, ignore some later occurring contextual information within the word that specified rate and lexical status. Importantly, however, they could not ignore speaking rate entirely. Although they could base their decision on only the early portion of the word, when doing so they treated the word as if it were physically short- that is to say, as if there were no later occurring information specifying a slower rate. This suggests that listeners always take account of rate when identifying the voicing value of a consonant, but precisely which information within the word is used to specify rate can vary with task demands. PMID- 2971768 TI - Central and peripheral representation of whispered and voiced speech. AB - Whispered speech is very different acoustically from normally voiced speech, yet listeners appear to have little trouble perceiving whispered speech. Two selective adaptation experiments explored the basis for the common perception of whispered and voiced speech, using two synthetic /ba/-/wa/ continua (one voiced, and one whispered). In the first experiment the endpoints of each series were used as adaptors, along with several nonspeech adaptors. Speech adaptors produced reliable labeling shifts of syllables matching in periodicity (i.e., whispered whispered or voiced-voiced); somewhat smaller effects were found with mismatched periodicity. A periodic nonspeech tone with short rise time (the "pluck") produced adaptation effects like those for /ba/. These shifts occurred for whispered test syllables as well as voiced ones, indicating a common abstract level of representation for voiced and whispered stimuli. Experiment 2 replicated and extended Experiment 1, using same-ear and cross-ear adaptation conditions. There was perfect cross-ear transfer of the nonspeech adaptation effect, again implicating an abstract level of representation. The results support the existence of two levels of processing for complex acoustic signals. The commonality of whispered and voiced speech arises at the second, abstract level. Both this level, and the earlier, more directly acoustic level, are susceptible to adaptation effects. PMID- 2971769 TI - Stress in time. AB - The goals of this research were to determine whether speakers adjust the stress patterns of words within sentences to create an alternation between strong and weak beats and to explore whether this rhythmic alternation contributes to the characteristics stress differences between two major lexical categories of English. Two experiments suggested that speakers do alter lexical stress in accordance with rhythmic biases. When speakers produced disyllabic pseudowords in sentence contexts, they were more likely to place stress on the first syllable when the pseudoword was preceded by a weak stress and followed by a strong one than when the strong stress preceded and the weak followed. This occurred both when the pseudowords served as nouns and when they served as verbs. Text analyses further revealed that weakly stressed elements precede nouns more often than verbs, whereas such elements follow verbs more often than nouns. Thus, disyllabic nouns are more likely than disyllabic verbs to occupy contexts biased toward trochaic rhythm, a finding consistent with leftward dominant stress in disyllabic English nouns. The history of stress changes in English nouns and verbs also conforms with the view that rhythmic context may have contributed to the evolution of stress differences. Together, the findings suggest that the citation stress patterns of words may to some degree reflect adaptations of lexical knowledge to conditions of language performance. PMID- 2971770 TI - Haptically perceiving the distances reachable with hand-held objects. AB - Nine experiments are reported on the ability of people to perceive the distances reachable with hand-held rods that they could wield by movements about the wrist but not see. An observed linear relation between perceived and actual reaching distances with the rods held at one end was found to be unaffected by the density of the rods, the direction relative to the body in which they were wielded, and the frequency at which they were wielded. Manipulating (a) the position of an attached weight on an otherwise uniformly dense rod and (b) where a rod was grasped revealed that perceived reaching distance was governed by the principal moment(s) of inertia (I) of the hand-rod system about the axis of rotation. This dependency on moment of inertia (I) was found to hold even when the reaching distance was limited to the length of rod extending beyond an intermediate grasp. An account is given of the haptic subsystem (hand-muscles-joints-nerves) as a smart perceptual instrument in the Runeson (1977) sense, characterizable by an operator equation in which one operator functionally diagonalizes the inertia and strain tensors. Attunement to the invariants of the inertia tensor over major physical transformations may be the defining property of the haptic subsystem. This property is discussed from the Gibsonian (ecological) perspectives of information as invariants over transformations and of intentions as extraordinary constraints on natural law. PMID- 2971771 TI - Programming saccadic eye movements. AB - This article addresses questions about the preparatory processes that immediately precede saccadic eye movements. Saccade latencies were measured in a task in which subjects were provided partial advance information about the spatial location of a target fixation. In one experiment, subjects were faster in initiating saccades when they knew either the direction or amplitude of the required movement in advance compared to a condition with equal uncertainty about the number of potential saccade targets but without knowledge of the parameters required to execute the movement. These results suggest that the direction and amplitude for an upcoming saccade were calculated separately, and not in a fixed serial order. In another experiment, subjects appear to have programmed the saccades more holistically--with computations of direction and amplitude parameters occurring simultaneously. The implications of these results for models of eye movement preparation are discussed. PMID- 2971772 TI - Tests of a temporal theory of attentional binding. AB - Different features of stimuli present in the field of view appear to be registered in different cortical maps. How, then, are the features that come from the same object bound together rather than mistakenly assembled with features coming from other simultaneously present objects? One theory supposes that an attentional mechanism intercepts input coming from particular retinal locations at a way station prior to parsing of the features from the same object. Any enhancement (or facilitation) at that stage will cause all the features from that object to be modified simultaneously in the downstream registers. The imposed temporal synchronicity serves as the essential binding cue. Five experiments provided no support for the theory. There is no tendency for synchronicity of features to cause binding unless the features come from the same location. Location, rather than temporal synchronicity, appears to be the essential cue for binding. PMID- 2971773 TI - Components of the location probability effect in visual search tasks. AB - In visual search tasks, targets are detected more rapidly when they appear in locations that commonly contain a target than when they appear in locations that rarely contain a target. Five experiments were conducted to investigate two specific properties of this location probability effect: its dependence on spatial location versus relative position and its dependence on or independence of target identity. In Experiment 1 spatial location of a stimulus row was varied to determine whether high location probability facilitates target detection in a particular location in visual space or a particular relative position within the row. Both were facilitated to approximately the same extent. In Experiment 2 an inducing target occurred with high probability in one of four display locations, and a test target occurred with equal probability in all four locations. Both targets were found more quickly in the high-probability location than in the other locations, but the advantage associated with targets in the high probability location was larger for the inducing target than for the test target. In Experiments 3-5 the correspondence between the components observed in Experiments 1 and 2 was examined. The overall pattern of results was compatible with a model in which the location probability effect is produced partly by an attentional spotlight, which facilitates processing of any stimulus appearing in a particular location in visual space, and partly by a network of position specific letter detectors, which facilitates detection of a particular letter in a particular relative position within a display. Models with flexible scanning strategies were also considered. PMID- 2971774 TI - Font regularity constraints on the process of letter recognition. AB - Strings of four unrelated letters were presented for subjects to identify, followed by a patterned mask and then a forced choice test of each letter position. In Experiment 1, the type style in the regular conditions was consistent--all of the letters were of a single type font--whereas in the mixed condition, each string contained letters from two type fonts. Compared with the mixed condition, accuracy in the regular conditions was higher overall and increased at a faster rate as a function of processing time. This held across four sessions. In Experiment 2, the font in the mixed condition was varied either between or within letter strings; sizeable advantages for the regular conditions were found with both mix-methods. The results are consistent with the idea of a schemalike perceptual system that becomes tuned to the regularities of a particular font in order to process visual information efficiently. PMID- 2971775 TI - Sequence influence on the organization of meaningless serial stimuli: economy after all. AB - This article provides evidence for an extension of structural information theory, a theory which describes perceptual organization formally, into a more general theory of representation that takes the role of organizations obtained earlier into account. In eight experiments, subjects study series of colored dots. Each series contained 6-8 dots of different colors, and viewing time was between 400 ms and 60 s. In several experimental procedures, subjects recalled the series presented. The experiments systematically varied the economy of the organizations of the series as described by structural information theory as well as a number of aspects of the context in which the series were presented, like order of presentation of a series, order between the series, viewing span, viewing time, and recall procedure. Recall was influenced both by the economy as well as by the context. It is shown how the context influences the relative strength of the most economical organization as compared with rival, local organizations. A structured network model is presented that accounts for the influence of both the economy and the context. PMID- 2971776 TI - "Ratio" and "difference" judgments for length, area, and volume: are there two classes of sensory continua? AB - Subjects were required to judge ratios and differences of (a) line length for pairs of lines, (b) area for pairs of squares, and (c) volume for pairs of cubes. Nonmetric analyses of these judgments indicated that all subjects were able to make consistent ratio judgments for all three continua. Many of the subjects, when asked to judge subjective differences, however, performed as if they were judging subjective ratios rather than differences. The data for the few subjects who appeared to be judging subjective differences were not consistent across subjects and conditions. Previous studies of ratio and difference judgments of loudness and heaviness, on the other hand, showed the opposite pattern, in that subjects most often behaved as if they were judging sensory differences when asked to judge sensory ratios. We propose that ratio judgments are more natural to perceptual continua along which stimuli are easily "decomposed" into a number of smaller perceptual units. PMID- 2971777 TI - Assessment of the taste interaction between two qualitatively similar-tasting substances: a comparison between comparison rules. AB - The taste interaction between sucrose and fructose was assessed by using three different comparison procedures: the summated response comparison, the factorial plot comparison, and the equimolar comparison rule. The perceived sweetness intensities were obtained on a ratio scale by using a functional measurement approach in combination with a two-stimulus procedure. The conclusions obtained from each of the three comparison rules were identical. The taste interaction between sucrose and fructose could be explained to a large extent, but not completely, by the apparent taste "interactions" within sucrose and fructose as single substances. It is argued that the apparent taste interaction within a large number of single sugars and between two of these sugars in a mixture is somewhat synergistic at low sweetness levels, additive at intermediate sweetness levels, and suppressive at high sweetness levels. PMID- 2971778 TI - A warning about median reaction time. AB - When used with positively skewed reaction time distributions, sample medians tend to over-estimate population medians. The extent of overestimation is related directly to the amount of skew in the reaction time distributions and inversely to the size of the sample over which the median is computed. Simulations indicate that overestimation could approach 50 ms with small samples and highly skewed distributions. An important practical consequence of the bias in median reaction time is that sample medians must not be used to compare reaction times across experimental conditions when there are unequal numbers of trials in the conditions. If medians are used with unequal sample sizes, then the bias may produce an artifactual difference in conditions or conceal a true difference. Some recent studies of cuing and stimulus probability effects provide examples of this potential artifact. PMID- 2971779 TI - Identification of a phosphoglucomutase 1 (PGM 1) variant in a case of murder/suicide. AB - A murder/suicide case is reported in which a phosphoglucomutase (PGM) 1*W9 variant was detected in a woman, her child, and from blood collected at the scene. PMID- 2971780 TI - Phosphoglucomutase and haptoglobin phenotypes among Australian Caucasians. PMID- 2971781 TI - Experimental study of argon-dye laser photoradiation therapy on tumor-bearing mice. PMID- 2971782 TI - Cutaneous sporotrichosis. PMID- 2971783 TI - Involvement of norepinephrine neurons in the hypothermia induced by intracerebroventricular administration of 6-hydroxydopamine in mice, evidenced by antidepressants. AB - The intracerebroventricular (i.c.v.) administration of increasing doses of 6 hydroxydopamine (6OHDA) (12.5-50 micrograms) induces in mice a dose-dependent hypothermic effect. This hypothermic effect is not affected either by serotonin uptake inhibitors (indalpine, clomipramine, trazodone, fluoxetine) or by dopamine uptake inhibitors (GBR 12783, amineptine). On the contrary, the hypothermia is partly antagonized by norepinephrine uptake inhibitors (desipramine, nomifensine, viloxazine, maprotiline, protryptiline), as well as amfonelic acid. The antagonism elicited by desipramine is observed when the drug is administered intraperitoneally (from 5 mg/kg) or intracerebroventricularly (from 5 microgram per mouse). 6-hydroxydopamine-induced hypothermia is antagonized by imipramine after a time lag of 1 hour; this antagonism lasts 6-11 hours after intraperitoneal administration of the drug (20 mg/kg). The hypothermic effect of 6-hydroxydopamine is diminished by a previous 6-hydroxydopamine i.c.v. administration (50 micrograms, 7 days before), except in mice pretreated with desipramine at the time of the first 6-hydroxydopamine injection. The hypothermic effect is completely abolished by two previous 6-hydroxydopamine i.c.v. administrations (50 micrograms, 7 days interval). It is also decreased in mice receiving DSP4 15 days before testing (50 mg/kg, i.p.). Finally, neither haloperidol (0.5 mg/kg i.p.) nor SCH23390 (100 micrograms/kg s.c.) antagonize 6 hydroxydopamine-induced hypothermia. It is concluded that this effect is largely depending on central norepinephrine neurons. PMID- 2971784 TI - Dissociation kinetics of [3H]paroxetine binding to rat brain consistent with a single-site model of the antidepressant binding/5-hydroxytryptamine uptake site. AB - The effects of 5-hydroxytryptamine (5-HT) and 5-HT uptake inhibitors on the dissociation of [3H]paroxetine from rat brain membrane binding sites have been investigated. The dissociation induced by 5-HT (100 microM), paroxetine (0.15 microM), clomipramine (1 microM), citalopram (1 microM), imipramine (1 microM), or norzimeldine (1 microM) was consistent with first-order dissociation kinetics with half-life values of dissociation (t1/2) between 130 and 140 min. The dissociation induced by the combination of 5-HT (100 microM) with either citalopram (1 microM) or imipramine (1 microM) was not different from that initiated by either agent alone. These dissociation data, which are at variance with previous data on the 5-HT transporter labeled with [3H]imipramine, support a single-site model of the antidepressant binding/5-HT uptake site. PMID- 2971785 TI - Morphometric demonstration of atrophic changes in the cerebral cortex, white matter, and neostriatum in Huntington's disease. AB - We performed morphometric analysis of five standardized coronal brain slices at anterior frontal (AF), caudate-putamen-accumbens (CAP), globus pallidus (GP), lateral geniculate nucleus (LGN), and parieto-occipital fissure (OCP) levels in 30 patients with Huntington's disease (HD) and 13 controls. Associated with the 30% mean reduction in brain weight in HD patients (p less than 0.001) were significantly smaller overall cross-sectional areas of brain at all five levels studied, with striking losses in cerebral cortex (21-29%), white matter (29-34%), caudate (57%), putamen (64%), and thalamus (28%) (all p less than 0.005). In addition, the ventricular system was dilated up to 2.5 times normal at CAP, GP, and LGN levels, 9.5 times normal at the OCP level, and 13 times normal at the AF level. Higher grades of severity of HD had greater reductions in the cross sectional area of the caudate, putamen, thalamus, and cerebral cortex (p less than 0.005-0.001), and larger ventricles (p = 0.08) compared to lower (less severe) grades of HD. The findings confirm and quantitate the severe atrophy of the neostriatum, in addition to demonstrating a severe loss of cerebral cortex and subcortical white matter in HD. The global atrophy of cerebral cortex and white matter observed in all degrees of HD may account for the cognitive and neuropsychiatric impairments which often precede the onset of chorea. PMID- 2971786 TI - Laminin and heparan sulphate proteoglycan in the lesioned adult mammalian central nervous system and their possible relationship to axonal sprouting. AB - Several in vitro studies indicate that the extracellular matrix (ECM) glycoprotein laminin can promote neurite outgrowth from CNS (central nervous system) neurons. Laminin has been detected immunohistochemically in astrocytes in the embryonic but not the uninjured adult mammalian CNS. In the injured adult CNS, it is found in some reactive astrocytes located near the site of CNS lesions. In the present study, we have attempted to examine the relationship between these laminin+ astrocytes and the axonal sprouting that occurs after CNS injuries. This was studied in the intracranially transected adult rat optic nerve which consists of a cranial segment devoid of all retinal ganglion cell axons, and a retinal segment attached to the retina which contains some viable axons that undergo sprouting. Laminin+ reactive astrocytes were found in the cranial segment, but not in the retinal segment. In addition, the cut ends of the retinal and cranial segments were capped by an intensely laminin+, glial fibrillary acidic protein negative (GFAP) region. Axonal sprouts from the transected retinal ganglion cell axons, identified by anterogradely transported rhodamine isothiocynate (RITC), were confined to laminin-, GFAP+ regions of the retinal segment. These results suggest that injury-induced axonal sprouting in the adult mammalian CNS in vivo may be promoted by molecules other than laminin, that may be associated with astrocytes. The presence of heparan sulphate proteoglycan HSP G was also examined in the transected optic nerve because the neurite outgrowth promoting factors found in conditioned media derived from several cell types in vitro have been shown to consist of a complex of laminin and heparan sulphate proteoglycan. No significant changes in heparan sulphate proteoglycan-like immunoreactivity was observed after transection. The presence of laminin and HSPG were also examined in the lesioned adult rat cerebral cortex. PMID- 2971787 TI - Fluctuations in spontaneous discharge of hippocampal theta cells during sleep waking states and PCPA-induced insomnia. AB - 1. The spontaneous activity of hippocampal theta cells in head-restrained cats was recorded during slow-wave sleep (SWS), paradoxical sleep (PS), and the attentive state of bird watching (BW). We also recorded theta cell activity during a state of insomnia with pontogeniculoocipital (PGO) waves, which was induced by the administration of p-chlorophenylalanine (PCPA), a selective inhibitor of serotonin synthesis. 2. The time-dependent structure of fluctuations in theta cell activity was evaluated by power spectral analysis and Markovian analysis. The coefficient of variation for these time series was used as a measure of the variability of theta cell activity, which indicates the relative amplitude of fluctuations. 3. During SWS, theta cell activity showed a larger variability and a flat spectrum, i.e., low Markovian properties. During PS, this activity exhibited smaller variability and high spectral density in a low frequency band (0.01-1.0 Hz), i.e., high Markovian properties. During BW, variability, spectral and Markovian properties were intermediate. 4. The firing pattern of theta cells during PCPA-induced insomnia was similar to that during PS. However, after the administration of either a serotonin agonist, 5-Methoxy N,N-dimethyltryptamine, or a choline antagonist, atropine sulfate, theta cell activity no longer exhibited PS-like fluctuations, revealing instead a firing pattern similar to that during SWS. 5. During PS and PCPA-induced insomnia, not only the unit activity of theta cells, but PGO activity and theta wave frequency exhibited slow fluctuations, i.e., the high spectral density in the low-frequency band (0.01-1.0 Hz). 6. Cross-correlation analyses were performed between the fluctuations in theta cell activity, theta wave frequency, and PGO activity. These fluctuations correlated with each other during both PS and PCPA-induced insomnia. Bursts of PGO waves especially contributed to these cross-correlations. 7. These results suggest, first, that the slow fluctuations of the theta cell activity during PS and PCPA-induced insomnia is the physiological expression of the removal of aminergic influences and, secondly, that they are dependent on cholinergic mechanisms, including PGO generators. PMID- 2971788 TI - Abnormal captopril renogram with a technetium-99m-labeled hippuran analog. AB - A case of renovascular hypertension is presented in which the [131I]hippuran renogram was initially normal, but became strikingly abnormal upon administration of the angiotensin converting enzyme (ACE) inhibitor captopril. The patient presented with fibromuscular dysplasia of the renal arteries, which was shown by hippuran renography to be functionally significant on the right side. She became normotensive after angioplasty of the right renal artery. Hypertension recurred a year later, at which time the renogram was normal without captopril, but showed functionally significant left renal artery stenosis with captopril challenge. Both the conventional agent, [131I]hippuran, and an experimental new 99mTc labeled hippuran analog, [99mTc]MAG3, were used. Angiography confirmed progression of disease on the left side, which was successfully treated by angioplasty. Functionally significant unilateral renal artery stenosis was thus demonstrated first on the right side and then, 1 yr later, on the left side, using hippuran and [99mTc]MAG3. Anatomic progression of disease was documented by angiography. PMID- 2971790 TI - Coping with back pain. PMID- 2971789 TI - Causes of false-negative auscultation of regurgitant lesions: a Doppler echocardiographic study of 294 patients. AB - Few data are available regarding the prevalence and causes of false-negative auscultation (mis-auscultation) of aortic (AR), mitral (MR), or tricuspid regurgitation (TR), and there are no such data that are relevant when the patient's pretest probability of having regurgitation is unknown. The authors therefore studied 294 patients examined by pulsed Doppler echocardiography. On 755 examinations (2.57 examinations per patient), Doppler velocity patterns typical of AR, MR, or TR were found in 63, 96, and 49 patients, respectively. For all three murmurs, mis-auscultation was the rule, rather than the exception, with sensitivities of auscultation ranging from 0 to 37%, depending (but weakly) on the site of the murmur and the years of training of the observer. Specificity of auscultation was high (85% to 100%). The factors associated with the mis auscultation of AR were poor image quality in the echocardiograms, absence of cardiomegaly, and less experience of the examiner. The probability of missing MR increased in the presence of coronary artery disease (CAD) or if the examiner had less experience. The likelihood of missing TR by auscultation was increased by CAD, obesity, chronic obstructive pulmonary disease, or the absence of cardiomegaly. This study suggests that there is a high prevalence of "silent" murmurs, and that not hearing a regurgitant murmur does not suffice to rule out the presence of regurgitation. PMID- 2971791 TI - Demonstration of type I and type II somatomedin receptors on bovine growth plate chondrocytes. AB - The chondrocytes of the epiphyseal growth plate are the presumed target cells for hormones regulating skeletal growth. The somatomedins, a family of low molecular weight peptides, are thought to play a stimulatory role in this regulation. The cellular actions of the somatomedins are themselves determined by binding to specific receptors on target cells. Previous studies have characterized a specific receptor for somatomedin-C (Sm-C) or insulin-like growth factor I (IGF I) on bovine growth plate chondrocytes (GPCs). We now report the characterization of a second type of somatomedin receptor on these cells that is more specific for another class of somatomedin represented by multiplication-stimulating activity (MSA) or rat insulin-like growth factor II (rIGF-II). Binding of [125I]MSA/rIGF II to isolated GPCs was time dependent and saturable. Unlabeled Mr 7,100 MSA/rIGF II and Sm-C/IGF-I were approximately equipotent in competing with [125I]MSA/rIGF II for binding, while Mr 8,600 MSA/rIGF-II was an order of magnitude less potent. Low levels of competition by insulin appeared in some studies at concentrations of 10(-7) M and higher, suggesting displacement of [125I]MSA/rIGF-II binding to the Sm-C/IGF-I receptor. In affinity-labeling studies, [125I]MSA/rIGF-I labeled a complex of Mr greater than 300,000 (unreduced) and of Mr 140,000 (reduced), consistent with a type I somatomedin receptor composed of disulfide-linked subunits. [125I]MSA/rIGF-II labeled a Mr 240,000 moiety (unreduced) and Mr 260,000 (reduced), consistent with a type II somatomedin receptor. Both affinity labeling and kinetic data revealed cross-binding of MSA/rIGF-II and insulin with the type I receptor and of Sm-C/IGF-I with the type II receptor. In contrast, the type II receptor did not recognize insulin. These data suggest a complex pattern of graded specificity of these receptors for their ligands. These data are consistent with the hypothesis that IGF-II as well as Sm-C/IGF-I participate in the stimulation of skeletal growth. PMID- 2971792 TI - Ischemic bowel after primary closure for gastroschisis. AB - Between 1983 and 1986, four newborns who had primary closure of gastroschisis had postoperative ischemic bowel. Suspicion was raised almost immediately after closure that something was wrong inside the abdomen when there was persistent acidosis, sepsis, abdominal wall redness, and a generalized worsening condition. All four neonates were re-explored. Necrotic bowel was found, and three required silon pouch closure. The two survivors were left with a temporary short gut. Whether the cause of the bowel ischemia in the four babies was due to excessive intraabdominal pressure, volvulus, or the intestines being too vigorously manipulated, is speculative. Therefore, excessive manipulation and compression of gastroschisis contents seem unwise; if such a newborn has persistence of the above signs and symptoms, immediate reoperation and decompression are warranted. PMID- 2971793 TI - Metabolic reduction of acetohexamide in rat kidney: sex difference and effect of streptozotocin-induced diabetes. AB - The acetohexamide reductase activity in 10000 x g supernatant fluids of kidney homogenates was significantly higher in male than in female rats. Although difference in activity of acetohexamide reductase in the cytosol between the sexes was not observed, the activity in the microsomes was considerably higher in male than in female rats. These findings indicate that the microsomal enzyme plays an important role in the sex difference of acetohexamide reduction by 10000 x g supernatant fluids of kidney homogenates. The sensitivities to inhibitors of microsomal acetohexamide reductase were different from those of cytosolic acetohexamide reductase. Furthermore, streptozotocin-induced diabetes significantly decreased acetohexamide reductase activity only in the kidney microsomes of male rats, resulting in the abolishment of the sex difference of acetohexamide reduction by 10000 x g supernatant fluids of kidney homogenates. PMID- 2971794 TI - Inhibition of intestinal alpha-glucosidase and postprandial hyperglycemia by N substituted moranoline derivatives. AB - The inhibitory activity of N-substituted derivatives of moranoline (1 deoxynojirimycin) against intestinal alpha-glucosidase and postprandial hyperglycemia was evaluated. None of the N-substituted derivatives studied displayed more potent inhibition of sucrase or maltase from rabbit intestines than the parent compound moranoline. However, unlike the in vitro results, many compounds exhibited more potent hypoglycemic activities than moranoline in sucrose-, or starch-loaded rat models. Alkenyl or aralkenyl derivatives displayed more potent hypoglycemic activities than alkyl or aralkyl derivatives. There was a weak correlation between in vitro and in vivo assay systems found by statistical analysis. It is necessary to evaluate compounds in vivo in order to select the most potent hypoglycemic compound. PMID- 2971795 TI - Changes in lipid peroxide concentrations in plasma and tissues by repeated administration of clioquinol to neonatal rats. AB - The changes in lipid peroxide concentrations in plasma and tissues after subcutaneous administration of clioquinol to clioquinol-sensitive (S-rats) and resistant neonatal rats (R-rats) were investigated. When a fixed dose of 150 mg/kg/d of clioquinol was given to R-rats for 14 d after birth, no significant difference in lipid peroxide concentrations in plasma, liver, kidney, brain and spinal cord at 5, 10 and 15 d was observed between clioquinol-treated and untreated rats. However, with increasing doses of clioquinol to R-rats every 5 d (150----300----600 mg/kg/d), the lipid peroxide concentrations at 15 d were higher in plasma, brain and spinal cord of clioquinol-treated rats than in those of untreated rats. These results suggested that repeated administrations of large doses of clioquinol to rats increased the lipid peroxides in nerve tissues. With S-rats at 5 d after birth, the lipid peroxide concentrations in liver were approximately twice those in R-rats regardless of the clioquinol administration. PMID- 2971796 TI - Effects of putative thromboxane receptor agonists and antagonists on rat small intestinal ion transport. AB - Effects of a thromboxane mimic, U46619, on electrolyte transport were examined in vitro using stripped segments of rat ileal mucosa mounted in Ussing chambers. Addition of U46619 to the serosal bathing solution elicited a transient increase in short-circuit current (Isc) and decrease in transepithelial conductance (Gt). The increase in Isc was accompanied by a transient increase in Cl- secretion and decrease in Na+ absorption. In the steady-state, Isc was not increased whereas Gt remained decreased and Na+ and Cl- absorption were inhibited. Removal of Cl- or pretreatment with serosal and mucosal indomethacin (1 microM) or the thromboxane receptor antagonist, SK&F 88046, added to the serosal bathing solution, inhibited the increase in Isc stimulated by U46619 (apparent KB approximately 8 nM). The effects of U46619 on both Isc and Gt are qualitatively similar to those resulting from stimulation with leukotriene D4. However, the changes in Isc with leukotriene D4 (10 microM) are antagonized by SK&F 88046 only at high concentrations (1-10 microM). In addition, the secretagogues prostaglandin F2 alpha, lys-bradykinin, serotonin and histamine, produce qualitatively similar changes in Isc to those seen with U46619 without altering Gt. With the exception of prostaglandin F2 alpha, the effects of these secretagogues are not inhibited by SK&F 88046 (10 microM). These results indicate that U46619 acts at a thromboxane receptor to stimulate intestinal Cl- secretion and inhibit Na+ and Cl absorption. These changes are inhibited selectively by the thromboxane receptor antagonist, SK&F 88046. PMID- 2971797 TI - Interactions of D1 and D2 dopamine receptors on the ipsilateral vs. contralateral side in rats with unilateral lesions of the dopaminergic nigrostriatal pathway. AB - In rats with a unilateral lesion of the nigrostriatal dopaminergic pathway, the ipsilateral rotation produced by the enhanced actions of endogenous dopamine (DA) on the nonlesioned side, induced by either the DA-releasing drug amphetamine or the DA uptake inhibitor GBR 13069, was blocked effectively by pretreatment with either the selective D1 DA receptor antagonist, SCH 23390, or the D2 selective antagonist, haloperidol. In contrast, contralateral rotation produced by apomorphine or I-dihydroxyphenylalanine, which lead to the preferential activation of D1 and D2 receptors on the lesioned side, was effectively prevented only when both receptor subtypes were inhibited. The results of these experiments demonstrate that the interaction between D1 and D2 receptors in the lesioned side differs from that in the nonlesioned side. Whereas the simultaneous stimulation of both DA receptor subtypes in the normally innervated basal ganglia is required for the production of turning behavior, the stimulation of either subtype alone in the dopaminergic denervated side can produce rotation. However, the concurrent administration of the D1 agonist, SKF 38393, with the D2 agonist, LY 171555, produced a synergistic effect on contralateral rotation. These results suggest that there is preservation of at least some functional interaction between D1 and D2 receptors in the lesioned basal ganglia but that there may be in addition a mechanism by which the two receptor subtypes can function independently of each other. The unilaterally lesioned rat appears to be a very good model in which to study the interaction between D1 and D2 receptors under conditions of both normal innervation and of DA denervation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971798 TI - [3H]SCH 23390 labels both dopamine-1 and 5-hydroxytryptamine1c receptors in the choroid plexus. AB - [3H]SCH 23390 has been used to label the D-1 subtype of dopamine receptors. Quantitative autoradiographic studies with [3H]SCH 23390 have revealed high levels of binding in many regions of rat brain including the caudate-putamen, nucleus accumbens, substantia nigra and choroid plexus. The selectivity of the binding of [3H]SCH 23390 was characterized further in studies using homogenates of canine choroid plexus. Scatchard analysis of the binding of increasing concentrations of [3H]SCH 23390 resulted in a curvilinear plot when (+) butaclamol was used to define specific binding. Nonlinear regression analysis of untransformed data was consistent with the presence of two distinct classes of binding sites. The high-affinity site (Kd, 0.4 nM) was present at low density (maximum binding sites, 30 fmol/mg of protein) and had the pharmacological properties expected of a D-1 receptor. The low-affinity site (Kd, 24 nM) was present at a much greater density (maximum binding sites, 350 fmol/mg of protein) and had the pharmacological properties expected of a 5-hydroxytryptamine1c receptor. Quantitative autoradiographic studies of the binding of [3H]SCH 23390 to sections of rat brain also suggested that 5-hydroxytroptamine1c receptors in the choroid plexus are labeled by [3H]SCH 23390. It is possible that [3H]SCH 23390 labels 5-hydroxytryptamine1c receptors in other brain regions as well. PMID- 2971799 TI - Stain potential of four microfilled composites. PMID- 2971800 TI - Effects of four lubricants used during incremental insertion of two types of visible light-activated composites. AB - The effect of alcohols, bonding resins, or dentin adhesives on the adhesive bond between the layers of visible light-activated composites when used as lubricant for composite instruments was investigated. The results of this study indicated that alcohols are not compatible with certain BIS-GMA-based and UEDMA-based composite systems (Herculite and Heliomolar composites were used for this investigation). Use of ethanol as a lubricant caused pure adhesive failure and low bond strength in UEDMA-based composite (Heliomolar), indicating that the bond strength was less than the cohesive strength of the composite. A bonding resin of a similar brand (or the same system) used to lubricate the composite instrument did not affect the bond strength of the bonded composite specimens. In light of this study, a bonding resin of the same resin system as the composite may be the most suitable and reliable material to be used for lubricating the composite instrument. PMID- 2971801 TI - Strength of composites repaired by laminating with dissimilar composites. PMID- 2971802 TI - Alloantigen presenting capacity of human decidual tissue. AB - The capacity of first trimester human decidua-derived cells to serve as accessory cells for the presentation of alloantigens to unprimed T lymphocytes was assessed using a mixed lymphocyte culture (MLC) between accessory cell-depleted responder and stimulator peripheral blood lymphocytes (d PBL). In all of the seven experiments performed decidua-derived cells achieved significant reconstitution of the lymphoproliferative response between autologous responder and stimulator dPBL. Reconstitution indices (RIs) ranged between 3.1 and 22.2 and were significant in all cases. In six out of the seven experiments the level of lymphoproliferation exceeded that between untreated responder PBLs co-cultured with stimulator d PBL. Substitution of recombinant interleukin 1 (rIL-1) alpha or beta for the decidua-derived cells resulted in significantly lower RIs than those achieved by the decidua-derived cells. These findings suggest that decidua derived cells can serve as antigen presenting cells for the presentation of membrane-bound alloantigens in a primary lymphoproliferative response. This may have implications for maternal recognition of fetal antigens during first pregnancy. PMID- 2971803 TI - Immunoregulation and prostaglandin production by mechanically-derived and enzyme derived murine decidual cells. AB - Uterine cells from pregnant mice release soluble mediators which suppress in vitro immune responses non-specifically. Two methodologic approaches have been employed to obtain regulatory cells, and, although all cell populations derived by both methods were immunosuppressive, different factors and possibly different mechanisms of suppression appear to have been involved. The current study was designed to determine how suppression by mechanically and enzymatically derived cells might differ. Both enzymatically and mechanically derived cell populations suppressed mixed lymphocyte reactions non-specifically, mediated their effects through release of soluble factors, were immunosuppressive whether added at initiation or near the end of culture, suppressed generation of both IL-1 and IL 2, and suppression was not reversible with IL-1, IL-2, gamma interferon or combinations of the above. Immunosuppression by enzymatically derived, but not mechanically derived cells, was reversible with indomethacin. Enzymatically derived cells, but not mechanically derived cells, produced considerable amounts of PGE2. Morphological and immunohistochemical analysis revealed significant numbers of macrophages in both populations. Enzymatic treatment of mechanically derived cells did not increase the output of PGE2 when cells were cultured in vitro. The results suggest that, despite several similarities, enzymatically and mechanically derived cell populations are distinct and produce a distinct profile of soluble suppressive molecules. PMID- 2971804 TI - The uterotubal junction. A proposal for classifying its morphology as assessed with hysteroscopy. AB - Hysteroscopy was used to characterize the morphology of the uterotubal ostium (UTO). A comparison was made with the laparoscopic assessment of the tube in 86 infertile women. In addition, histology was performed on the UTO from women undergoing hysterectomy for cervical dysplasia. These comparisons revealed that the incidence of mucosal damage to the UTO is not related to that of adnexal adhesions unless the postinflammatory changes are severe. Most notable was the correlation between endosalpingeal hyperplasia and salpingitis isthmica nodosa. PMID- 2971805 TI - Laparoscopic management of ovarian pregnancy. A case report. AB - Operative laparoscopy has been reported to be a useful alternative to laparotomy for the treatment of tubal pregnancy. Ovarian pregnancy is a rare form of ectopic gestation that can be treated with operative laparoscopy. The most serious problem in treating ovarian pregnancy is making an accurate diagnosis; pathologic documentation is the only way to confirm the diagnosis. Postoperative follow-up should include evaluation of the patient for bleeding or infection and serial monitoring of quantitative beta-human chorionic gonadotropin levels to confirm the complete removal of placental tissue. PMID- 2971806 TI - The role of plasminogen in interleukin-1 mediated cartilage degradation. AB - Explants of 35SO4 labelled rabbit articular cartilage, cultured for 3 days with either 5 X 10(4) rabbit peritoneal cells (PEC) or 1:10 macrophage conditioned medium (MCM), released 30-40% of labelled proteoglycans into the medium while controls released 8-12%. The addition of 1 mM 4 aminophenylmercuric acetate (APMA) or 0.2 U/ml plasminogen increased proteoglycan release to 85%. Similar results were obtained when recombinant human interleukin-1 (IL-1) was used instead of MCM. Further, supernatant from MCM stimulated chondrocytes, incubated with dead cartilage explants for 3 days, did not significantly increase proteoglycan release above the background level of cartilage alone (7-10%), nor did the addition of 5 X 10(4) PEC to cultures of dead cartilage explants plus supernatant from MCM stimulated chondrocytes make any significant difference, indicating that supernatant from MCM stimulated chondrocytes and PEC alone had negligible cartilage proteoglycan degrading activity in these experiments. The inclusion of 0.1 mM APMA or 0.2 U/ml plasminogen in cultures of dead cartilage explants plus supernatant from MCM stimulated chondrocytes, however, increased proteoglycan release to 80-93%, with or without PEC. Our results suggest that plasminogen, activated by a product from IL-1 stimulated chondrocytes, greatly enhanced IL-1 mediated cartilage degradation by activating latent metalloproteinases. PMID- 2971807 TI - T-lymphocyte subsets and delayed type hypersensitivity as classification parameters for HIV infection among drug-addicts. AB - HIV selectively infects T helper lymphocytes, which play an essential role in cell-mediated immunity mechanisms. Impairment of these mechanisms can be evaluated, other than by clinical parameters, by testing lymphocytic subsets and cutaneous delayed type hypersensitivity. All these indices are used in the Walter Reed Foundation (WR) staging system, which includes the different stages of HIV infection. In our study we investigated the above mentioned parameters in a series of 180 HIV positive PDAs. A classification of the same patients according to WR's criteria was also designed. Of our patients, 46.1% resulted normoergic, 27.2% hypoergic and 26.7% anergic. Of all normoergic subjects, 83.7% (with more than 400 OKT4+ cells/mmc) could be assigned to the WR2 and 16.3% (with less than 400 OKT4+ cells/mmc) to the WR3 staging group. Among the hypoergic patients, only 34.7% were assignable to the WR4 group, whereas the majority (65.3%) had more than 400 OKT4+ cell/mmc. Even among the anergic group, 62.5% did not fit the WR5 stage, exhibiting an OKT4+ level higher than 400/mmc. In the latter 2 groups (referred to as D and E), T helper lymphocyte counts were performed at regular intervals; however, only the ones followed-up for at least 6 months were evaluated. OKT4+ cells were shown to decrease more frequently and more rapidly in the anergic than in the hypoergic patients, which points to cutaneous anergy as representing an early sign of impairment of cell-mediated immunity. PMID- 2971808 TI - The occurrence of maladaptive health-related behaviors and teacher-rated conduct problems in children of chronic low back pain patients. AB - Chronic low back pain (CLBP) is often associated with a complex of behaviors in the patient which cause interference in adaptive functioning of both the patient and the patient's family. This study examined a group of children of CLBP patients, a control group of children of diabetic patients, and a general control group of children. The study supported the hypothesis that, as a group, children of CLBP patients would exhibit a higher frequency of behaviors hypothesized to be learned through observation of and interaction with a CLBP parent than would children in either of the control groups. Significant differences between the pain group and the control groups on each of the nine teacher-reported and child self-report measures were observed. Implications and limitations of the study are discussed. PMID- 2971809 TI - The Minnesota Multiphasic Personality Inventory (MMPI) Hysteria (Hy) scale: scoring bodily concern and psychological denial subscales in chronic back pain patients. AB - Chronic back pain patients were compared to two nonpain comparison groups on Minnesota Multiphasic Personality Inventory (MMPI) Hysteria (Hy) scores and scores on two Hy subscales: Bodily Concern and Psychological Denial. Pain subjects had significantly higher scores on the Bodily Concern Subscale and lower scores on the Psychological Denial Subscale than nonpain subjects with similar elevated Hy scores. However, pain subjects had Psychological Denial scores similar to those of subjects with normal MMPI profiles, despite significantly higher Hy scores. These findings are interpreted to support the hypothesis that, among pain patients, Hy elevations are partially accounted for by the endorsement of a disproportionate number of Bodily Concern items. Finally, within pain patients, scores on the Bodily Concern subscale were significantly related to more indices of pain duration and severity than were scores on the Psychological Denial subscale. The potential clinical utility of scoring these subscales is discussed. PMID- 2971810 TI - Chronic back pain and rheumatoid arthritis: predicting pain and disability from cognitive variables. AB - Cognitive-behavioral models of chronic pain emphasize the importance of situation specific as well as more general cognitive variables as mediators of emotional and behavioral reactions to nociceptive sensations and physical impairment. The relationship of situation-specific pain-related self-statements, convictions of personal control, pain severity, and disability levels was assessed in samples of chronic back pain and rheumatoid arthritis patients. Both the more general and the situation-specific sets of cognitive variables were more highly related to pain and disability than disease-related variables. This association was found in the back pain patients who displayed only marginal levels of organic findings as well as the rheumatoid arthritis sample who had a documented basis for their pain. The combination of both situation-specific and general cognitive variables explained between 32 and 60% of the variance in pain and disability, respectively. The addition of disease-related variables improved the predictions only marginally. These results lend support to the importance of cognitive factors in chronic pain syndromes. PMID- 2971811 TI - Late left ventricular function following angioplasty in acute myocardial infarction. PMID- 2971812 TI - Reactivation of the methylation-inactivated late E2A promoter of adenovirus type 2 by E1A (13 S) functions. AB - The inactivating effect of sequence-specific promoter methylations was extensively studied by using the late E2A promoter of adenovirus type 2 (Ad2) DNA. The modification of the three 5' CCGG 3' sequences at nucleotides +24, +6 and -215, relative to the cap site in this promoter, sufficed to silence the gene in transient expression either in Xenopus laevis oocytes or in mammalian cells, and after the fixation of the E2A promoter-chloramphenicol-acetyltransferase (CAT) gene construct in the genome of hamster cells. It will now be demonstrated that the inactivation of the late promoter of Ad2 DNA can be reversed by transactivating functions that are encoded in the 13S messenger RNA of the E1A region of Ad2 DNA. The reactivation of a methylation-inactivated eukaryotic promoter by transactivating functions has general significance in that the value of a regulatory signal can be fully realized only by its controlled reversibility. It was demonstrated in transient expression experiments that the 5' CCGG 3'-methylated late E2A promoter was at least partly reactivated in cell lines constitutively expressing the E1 region of Ad2 or of adenovirus type 5 (Ad5) DNA. The reactivation led to transcriptional initiation at the authentic cap sites of the late E2A promoter and was not associated with promoter demethylation, at least not in both DNA complements. Reactivation of the methylation-inactivated E2A promoter could also be demonstrated in two BHK21 cell lines (mc14 and mc20), which carried the late E2A promoter-CAT gene assembly in an integrated form. In these cell lines the late E2A promoter was methylated and the CAT gene was not expressed. By transfection of cell lines mc14 and mc20, the reactivating functions were shown to reside in the pAd2E1A-13 S cDNA clone of Ad2 DNA. The pAd2E1A-12 S cDNA clone or the pAd2E1B clone showed no reactivating function. These findings implicated the E1A 289 amino acid residue protein of Ad2, a well-known transactivator, as the reactivating function of the endogenous, previously dormant, late E2A promoter-CAT gene assembly. The methylated promoter was not demethylated, at least not in both complements, and it was shown that reactivation of the methylated promoter entailed transcriptional initiation at the authentic late E2A cap site. Since E1A and E1B jointly had a more pronounced effect, it was conceivable that genes in both regions acted together in the abrogation of the inhibitory effect of promoter methylations in the late E2A promoter. PMID- 2971813 TI - Effect of photoreactivation on mutagenesis of lambda phage by ultraviolet light. AB - There is disagreement in the literature as to whether the major mutagenic photoproduct induced in DNA by ultraviolet light is the cyclobutane dipyrimidine dimer, the most common product, or the [6-4] photoproduct, the next most frequent. In the experiments reported here, cyclobutane dimers were removed from irradiated lambda phage DNA by enzymatic photoreactivation, a process thought to affect no other photoproduct. Photoreactivation of lambda phage in host cells and of lambda DNA in solution reduced clear plaque mutants per plaque-forming unit by two-thirds, in host cells with a constant and near-maximal expression of the SOS functions required for mutagenesis. This result is interpreted to mean that removal of cyclobutane dimers in or near the mutated gene reduces mutation induced by ultraviolet light by two-thirds; therefore, cyclobutane dimers in the phage DNA are responsible for most observed mutations. DNA sequences of mutations in photoreactivated phage showed a smaller fraction of G.C to A.T transitions and a larger fraction of A.T to G.C transitions, compared to phage that were not photoreactivated. This suggests that cyclobutane dimers at TC and CC sites are particularly mutagenic. PMID- 2971814 TI - Immunocytochemical localization of natriuretic peptide sequences in the human right auricle. AB - Samples of the right auricle from ten patients undergoing surgery were processed for the immunocytochemical localization of atrial natriuretic peptide sequences (ANP-28 and the 26-92, 26-55 and 56-92 segments of the prohormone) using commercially available antisera and labelled with protein-A gold. Single antigen and double antigen (ANP-28 and one other) detection procedures were employed. The results show that there is no correlation between the ultrastructural appearance of specific heart granules and the molecular form of their ANP content. All specific granule types contain the full range of prohormone sequences. D granules are few and appear to be recently budded from the Golgi apparatus, whereas A and B profiles comprise the majority of specific granules and are more widely distributed in the cytoplasm. In the Golgi elements, peptide immunoreactivity is principally associated with the membrane but a proportion lies within the of vesicles and becomes concentrated within the core of developing granules. The density of ANP-immunoreactivity is higher in A profiles than in the other types. Multivesicular bodies are not intrinsically immunoreactive for ANP, nor are the numerous synaptic nerve profiles which are present in atrial muscle. In two elderly patients the basal lamina of myocytes was heavily labelled for ANP. PMID- 2971815 TI - Cellular oncogenes in cardiovascular disease. PMID- 2971816 TI - Understanding trimetrexate toxicity. PMID- 2971817 TI - Correlates of severe or life-threatening toxic effects from trimetrexate. AB - Trimetrexate, an investigational antifol, has been associated with marked variability in drug tolerance among patients. The agent is extensively protein bound, and hepatic biotransformation plays a major role in its elimination. In early phase II testing, nine of 15 patients who experienced life-threatening or fatal toxic effects from trimetrexate had albumin levels less than or equal to 3.5 g/dL prior to treatment. This prompted a review of the data base on 272 patients entered in phase I clinical trails. The incidence of severe or life threatening anemia, leukopenia, neutropenia, thrombocytopenia, mucositis, and hepatic toxic effects during the first course of trimetrexate was analyzed according to dose, schedule, prior treatment, and baseline protein and albumin levels. The schedules using doses given by short infusions of 30-60 minutes daily for 5 days or weekly for 3 weeks were generally associated with higher incidence of toxic effects than the schedules using doses given every other week by short infusions or those using continuous infusion. The occurrence of leukopenia and mucositis was dose related. Patients with baseline albumin levels less than or equal to 3.5 g/dL had higher incidence of all types of severe or life-threatening toxic effects than those with albumin levels greater than or equal to 3.6 g/dL, and the differences were significant for the development of anemia, thrombocytopenia, and mucositis. Similar correlations were noted for pretreatment protein levels less than or equal to 6.0 g/dL. The small cohort of patients with leukemia experienced substantial toxic effects and tended to have low protein and albumin levels. Performance status and prior therapy did not emerge as strong predictors of severe toxic effects in the univariate analysis. Multivariate analysis confirmed that the type of cancer (leukemia vs. solid tumor), dose, schedule, and baseline albumin level were significant and independent predictors of severe and life-threatening toxic effects in the phase I patient population. Multivariate analysis including only patients with solid tumors indicated that albumin level, dose, and schedule remained significant predictors of toxic effects. Since normal liver function as reflected by bilirubin and transaminase values were a requirement for eligibility, the results suggest that albumin and protein levels may provide a more sensitive index of hepatic function. Patients with hypoalbuminemia and hypoproteinemia are at increased risk of experiencing severe or life-threatening toxic effects from trimetrexate and should be treated cautiously. PMID- 2971818 TI - Trimetrexate: predictors of severe or life-threatening toxic effects. AB - In four phase II trials of trimetrexate given iv daily for 5 days, we noted marked variability among patients in the development of severe or life threatening toxic effects. In an effort to define which of 15 patient characteristics were associated with toxic effects of this degree, we have carried out single-factor and multifactor analyses on toxic effects during the first cycle of therapy in 68 patients. The final logistic regression model identified both low pretreatment serum protein levels and metastatic liver disease as significant correlates of severe toxic effects (P = .02 and P = .0005, respectively). While drug dose was an important element of the best logistic model, its statistical significance was only borderline. Trimetrexate is extensively protein bound and is cleared primarily by hepatic metabolism, so it is not unreasonable to believe that alteration in protein binding of the drug or in metabolic capacity of the liver could produce enhanced toxic effects. Although the validity of these predictors should be confirmed, it seems prudent to recommend lower starting doses of trimetrexate for patients with metastatic liver disease and/or low protein levels and dose escalation if toxic effects allow it. PMID- 2971819 TI - Effects of inhibitors of glycoprotein processing on oligodendroglial differentiation in primary cultures of embryonic rat brain cells. AB - The effects of N-linked oligosaccharide processing inhibitors on oligodendroglial differentiation were examined in cultures of embryonic rat brain cells. The glucosidase inhibitors, 1-deoxynojirimycin (dNM and castanospermine, were found to have marked inhibitory effects on the developmental expression of oligodendroglial properties, i.e., sulfogalactolipid synthesis and 2'3'-cyclic nucleotide 3'-phosphohydrolase (CNP). On the other hand, the mannosidase inhibitors, 1-deoxymannojirimycin (dMM) and swainsonine, had relatively little effects. Since both classes of inhibitors block the formation of complex-type oligosaccharide chains as revealed by concanavalin A-Sepharose affinity chromatography of the glycopeptides, complex oligosaccharides do not seem to play a role in oligodendroglial differentiation. The results indicate instead that the early trimming reactions involving the removal of glucose residues by processing glucosidases may be critical for the functioning of specific glycoprotein(s) essential to oligodendroglial differentiation. PMID- 2971820 TI - [Clinical experiences of occupational dermatoses in recent years]. PMID- 2971823 TI - Renovascular hypertension. PMID- 2971821 TI - Differential distribution of the adenovirus E1A proteins and colocalization of E1A with the 70-kilodalton cellular heat shock protein in infected cells. AB - Five distinct localization patterns were observed for the adenovirus E1A proteins in the nuclei of infected HeLa cells: diffuse, reticular, nucleolar, punctate, and peripheral. The variable distribution of E1A was correlated with the time postinfection and the cell cycle stage of the host cell at the time of infection. All staining patterns, with the exception of peripheral E1A localization, were associated with the early phase of infection since only the diffuse, reticular, nucleolar, and punctate staining patterns were observed in the presence of hydroxyurea. Because the E1A proteins (12S and 13S) stimulate the expression of the cellular heat shock 70-kilodalton protein (hsp70), we examined the intracellular distribution of hsp70 in the adenovirus-infected cells. Whereas hsp70 was predominantly cytoplasmic in the cells before infection, after adenovirus infection most of the protein was now found within the nucleus. Specifically, hsp70 was found within the nucleoli as well as exhibiting reticular, diffuse, and punctate nuclear staining patterns, analogous to those observed for the E1A proteins. Double-label indirect immunofluorescence of E1A and hsp70 in infected cells demonstrated a colocalization of these proteins in the nucleus. Translocation of hsp70 to the nucleus was dependent upon both adenovirus infection and expression of the E1A proteins. The localization of hsp70 was unaltered by infection with an E1A 9S cDNA virus which does not synthesize a functional E1A gene product. Moreover, the discrete nuclear localization patterns of E1A and the colocalization of E1A with hsp70 were not observed in adenovirus-transformed 293 cells which constitutively express E1A and E1B. E1A displayed exclusively diffuse nuclear staining in 293 cells; however, localization of E1A into the discrete nuclear patterns occurred after adenovirus infection of 293 cells. Immunoprecipitation of labeled infected-cell extracts with a monoclonal antibody directed against the E1A proteins resulted in precipitation of small amounts of hsp70 along with E1A. These data indicate that the adenovirus E1A proteins colocalize with, and possibly form a physical complex with, cellular hsp70 in infected cells. The relevance of this association, with respect to the function of these proteins during infection and the association of other oncoproteins with hsp70, is discussed. PMID- 2971824 TI - Simultaneous aortic reconstruction and bilateral renal revascularization. Is this a safe and effective procedure? AB - Between 1982 and 1987, 32 patients with severe aortorenal atherosclerosis had simultaneous aortic and bilateral renal revascularization. All patients were hypertensive. Eighteen patients (56%) had renal insufficiency with a mean serum creatinine (SC) of 2.8 mg/dl. Nine patients had an aortic aneurysm; the remaining 23 patients had aortoiliac occlusive disease of varying severity. Aortic reconstruction was done with either a straight (six patients) or bifurcated (26 patients) Dacron graft. Renal revascularization was accomplished with either bypass (60 arteries) or transaortic endarterectomy (four arteries). One patient died of pulmonary embolism (operative mortality rate 3%). Beneficial blood pressure response was achieved in 28 of 31 survivors, (90%). Among the 18 patients with renal insufficiency, mean SC was 2.80 +/- 1.18 mg/dl preoperatively and 1.65 +/- 0.48 mg/dl postoperatively (p less than 0.001). Among eight patients with severe renal dysfunction before surgery (SC greater than 3 mg/dl), mean SC was 3.90 +/- 0.85 mg/dl before and 1.79 +/- 0.69 mg/dl after operation (p less than 0.001). In follow-up extending to 58 months (mean 27.6 months), five late deaths occurred; cumulative survival was 94% at 2 years and 60% at 4 years. There were no instances of worsening hypertension; one patient had deteriorating renal function. These results indicate that severe aortorenal atherosclerosis can be managed with simultaneous aortic reconstruction and bilateral renal revascularization at low operative risk. In addition, there can be high expectation of significant and persisting benefit in both hypertension and renal dysfunction after operation. PMID- 2971825 TI - Surgical renal artery reconstruction after percutaneous transluminal angioplasty. AB - Percutaneous transluminal renal angioplasty (PTRA) is a controversial treatment for renal artery stenosis. This article discusses whether or not a prior attempt at PTRA compromises a subsequent elective or emergent surgical revascularization. Thirteen patients had surgical renal artery reconstruction after one or more PTRAs. Eight of the patients were treated because of atherosclerotic renal artery disease whereas five had a form of fibromuscular dysplasia. Five patients had renal artery injury directly related to the angioplasty. Four of these kidneys were saved. Eight patients were treated from 6 to 920 days after PTRA because of recurrent stenosis or occlusion of the renal artery. Only one of these kidneys was lost, an attempt at revascularization of a small kidney that failed to resume function. A prior attempt at PTRA did not compromise the ability of subsequent surgical revascularization to ameliorate hypertension. We conclude that surgical renal revascularization is not made less likely to succeed by a previous attempt at PTRA; even if the renal artery is thrombosed or perforated during the procedure, a reasonable chance of renal salvage is obtained by immediate surgical revascularization. PMID- 2971822 TI - Evaluation of the interaction of phi X174 gene products E and K in E-mediated lysis of Escherichia coli. AB - Gene K of bacteriophage phi X174 was cloned, and its gene product was localized in the cell envelope of Escherichia coli. Compared with the sole expression of the phi X174 lysis gene E, the simultaneous expression of the K and E genes had no effect on scheduling of cell lysis. Therefore, a direct interaction of proteins E and K could be excluded. In contrast, phi X174 infection of a host carrying a plasmid expressing gene K resulted in a delayed lysis and an apparent increase in phage titer. PMID- 2971826 TI - The Kensey catheter: preliminary results with a transluminal atherectomy tool. AB - The Kensey dynamic angioplasty instrument is an atherectomy device approved by the Food and Drug Administration that uses a rotating cam tip housed within a flexible polyurethane catheter to recanalize obstructed and stenotic arteries. Twenty patients with significant femoral arteriosclerotic occlusive disease underwent attempted transluminal endarterectomy of 23 extremities with the Kensey catheter. Significant improvements of superficial femoral artery luminal diameter was achieved in 10 of 13 patients with stenosis and passage of the spinning catheter tip at 60,000 to 90,000 rpm through areas of complete occlusion was successful in 4 of 10 cases. Balloon dilatation was used as an adjunct to increase the diameter of the superficial femoral artery lumen in 11 of 14 successful cases. This preliminary report provides technical data and short-term follow-up of this new innovative vascular tool. PMID- 2971827 TI - Efficacy of a mass hepatitis B vaccination program in Taiwan. Studies on 3464 infants of hepatitis B surface antigen-carrier mothers. AB - To evaluate the efficacy of the mass hepatitis B vaccination program in Taiwan in interrupting perinatal hepatitis B virus transmission, 3464 randomly selected 18 month-old infant vaccinees born to hepatitis B surface antigen-carrier mothers were recruited from 9697 eligible infants during a six-month period of the program. They were divided into ten groups according to maternal infectivity and compliance with the vaccination schedule. Serum samples were tested for hepatitis B surface antigen, antibody to hepatitis B surface antigen, and antibody to hepatitis B core antigen. In 786 infants who had highly infectious mothers and who received hepatitis B immune globulin and vaccine on schedule, the protective efficacy was about 85%. The efficacy seemed to be slightly lower in those immunized off schedule. Overall, 11% of infants still carried hepatitis B surface antigen, and 81% of the infants had antibody to hepatitis B surface antigen that exceeded 10 mIU/mL in more than 90% of them. The geometric mean titers of antibody to hepatitis B surface antigen were more than 200 mIU/mL in every group of infants. We conclude that the mass vaccination program is efficacious in preventing perinatal hepatitis B virus transmission and the chronic carrier state; most infant vaccinees have adequate levels of protective antibody at 18 months of age. This program is extremely significant in the control of hepatitis B virus infection in Taiwan. PMID- 2971828 TI - Experimental studies on ischemic injury and reperfusion injury to the cardiac sarcoplasmic reticulum--the myocardial protective effect of nicorandil. AB - Ischemic and reperfusion injuries to the myocardium were evaluated by measuring cardiac function and the calcium binding capacity of the sarcoplasmic reticulum (SR) and by studying the myocardial protective effect of nicorandil. While undergoing cardiopulmonary bypass, dogs were subjected to 120-min global myocardial ischemia and then to 120-min reperfusion. Group I hearts were arrested with untreated potassium cardioplegic solution; Group II with the same solution containing 2 mg/L and Group III with the same plus 10 mg/L of nicorandil. Group III exhibited better recovery from ischemic and reperfusion injuries than Group I with recovery rates of LV dp/dt max (95.0 +/- 28.9% vs 61.1 +/- 30.4, p less than 0.05) and LV negative dp/dt max (69.0 +/- 12.5% vs 46.8 +/- 21.7, p less than 0.05). The 3 groups showed a marked decrease in the max calcium binding capacity during ischemia compared with the Control Group but exhibited no further decrease after reperfusion. After ischemia and reperfusion, Group III (30.4 +/- 9.13 nmol Ca/mg protein, 30.0 +/- 8.50) demonstrated a significantly higher binding capacity than Group I (17.0 +/- 2.41 and 18.3 +/- 1.01, p less than 0.05), while Group II did not. These results suggest that ischemia is more injurious to the calcium binding capacity of SR than reperfusion and that 10 mg/L of nicorandil added to the cardioplegic solution preserves SR function and enhances the recovery of cardiac function. PMID- 2971829 TI - [Hepatitis B virus. Application of recombinant DNA technics to the development of hepatitis B vaccines and its diagnosis]. PMID- 2971830 TI - [Studies on immune deficiency in chronic renal failure. I. Lymphocyte subset and ConA-induced suppressor cell activity in chronic hemodialysis patients]. PMID- 2971831 TI - Possible involvement of sodium pump in the relaxation of rat aorta induced by alpha-human atrial natriuretic polypeptide. AB - In order to clarify whether the sodium handling of smooth muscle is associated with the relaxing action of alpha-human atrial natriuretic polypeptide (alpha hANP), we examined the sodium pump-related effects of alpha-hANP on rat aortic smooth muscles. Application of Ca2+ (1.0 to 10.0 mM) to the muscle preincubated in Ca2+-free, and K+-free or 0.5 mM K+ medium for 60 min induced a contraction. Pretreatment with alpha-hANP (1 x 10(-8) M) decreased the contraction evoked in 0.5 mM [K+]o but not that in K+-free medium. After a contraction was elicited by norepinephrine in K+-free solution, an addition of KCl (1.4-5.4 mM) caused a transient relaxation in a concentration-dependent manner, presumably due to the activation of electrogenic Na pump. The alpha-hANP enhanced the relaxation, which was sensitive to ouabain, and the potentiation by alpha-hANP was inversely related to the concentration of K+ added. When alpha-hANP was applied to relax the muscle precontracted by norepinephrine in the varied concentration of external K+, alpha-hANP-induced relaxation was greater in 1.4 or 2.7 mM [K+]o than in 0 or 5.4 mM [K+]o. These results suggest that the vasodilating effect of alpha-hANP is at least partially mediated by the activation of electrogenic Na, K pump and this effect is prominent when the Na, K-pump is partially suppressed. PMID- 2971832 TI - [Spontaneous diabetes and diabetes induced by streptozotocin in the rat and mouse]. PMID- 2971833 TI - [Animal models of non-insulin-dependent diabetes induced in the rat by experimental reduction of B cell mass]. PMID- 2971834 TI - Coronary angioplasty after thrombolysis. PMID- 2971835 TI - Atrial natriuretic peptide and furosemide effects on hydraulic pressure in the renal papilla. AB - Atrial peptides (ANP) have been shown to preferentially increase blood flow to juxtamedullary nephrons and to augment vasa recta blood flow. To determine the effect of this alteration in intrarenal blood flow distribution on pressure relationships in inner medullary structures and their significance as a determinant of ANP-induced natriuresis, we measured hydraulic pressures in vascular and tubule elements of the renal papilla exposed in Munich-Wistar rats in vivo during an euvolemic baseline period and again during an experimental period. Rats in Group 1 received intravenous infusion of rANP administered as a 4 micrograms/kg prime and 0.5 microgram/kg/min continuous infusion, and were maintained euvolemic by plasma replacement. Infusion of ANP resulted in significant natriuresis, diuresis and increase in inulin clearance. Within 90 seconds of initiation of this systemic infusion, vasa recta hydraulic pressures were markedly increased and exceeded the small pressure increment occurring in loops of Henle and collecting ducts. Infusion of furosemide in Group 2 rats at a dosage which reproduced the increase in urine flow in Group 1 was associated with small and equivalent increases in both vascular and tubule elements, indicating that the differential pressure response observed in Group 1 was not due to increased tubule fluid flow rates, but was rather a specific ANP-induced vascular effect. Group 3 rats received an infusion of ANP in a setting where its whole kidney hemodynamic effects were prevented. This resulted in a marked blunting of natriuresis and diuresis, and obliteration of the pressure differential between vasa recta and tubules observed in Group 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971836 TI - Determinants of immune complex-mediated glomerulonephritis. AB - We have studied the influence of steric factors on the clinico-pathologic expression of immune complex-mediated glomerular diseases, utilizing ferritin as an exogenous antigen. The tracer was planted in the left kidney either in the subepithelial layer of the glomerular capillary wall or on the endothelium and lamina rara interna. Subepithelial immune complex formation resulted in non inflammatory injury with heterologous and autologous proteinuric phases (115 +/- 16 mg/24 hrs on day 2; 183 +/- 16 mg/24 hrs on day 9) lasting four to five weeks. The glomerular filtration rate of the experimental left kidney was reduced by 19% at day 3, and was increased by 20% at day 12 over right kidney values. Immune complexes persisted for more than seven weeks in the lamina rara externa. In contrast, immune complex deposition on the endothelium and in the lamina rara interna led to acute transient anuria, with a 38% drop in glomerular filtration rate at one hour, massive platelet accumulation, followed by a strong inflammatory response. Proteinuria did not develop. Functional and structural integrity was restored within 24 hours, with complete clearing of immune deposits. We conclude that the distribution of exogenous antigens within the capillary wall determines the structural and functional expression of immune mediated glomerular diseases. PMID- 2971837 TI - [McVay's hernioplasty]. PMID- 2971838 TI - Non-responsiveness to hepatitis-B vaccination: revaccination and immunogenetic typing. AB - The variation in immune responses to standard inoculation of the hepatitis-B virus vaccine suggest that host factors influence response in ways that are not presently understood. We studied 25 low/nonresponding health care workers (anti HBs titer less than 50 IU/l) after the third inoculation of an experimental hepatitis-B vaccine to determine their immune status (through lymphocyte phenotypes) and HLA type. After application of a fourth inoculation, the seroconverting subjects showed only low anti-HBs levels; three male subjects remained anti-HBs negative. Twelve months after the fourth inoculation only 9 of 25 subjects (36%) maintained anti-HBs titer greater than 10 IU/l. Almost all subjects had normal B-cell and CD-4 and CD-8 counts and ratios. Relative to other European populations HLA-A-10 (P less than 0.05), B-12 (P less than 0.025), CW-5 (P less than 0.05), DR-3 (P less than 0.025), and DR-5 (P less than 0.025) were increased, whereas DR-2 (P less than 0.05) was decreased. However, after correction of the P-values for the number of HLA antigens determined, these differences were no longer significant. Furthermore, these HLA types were not the same as those reported in other studies (except for DR-3). We suggest that larger sample sizes or even not yet available immunogenetic markers will be required to prove an "immunogenetic background" in low/nonresponders, if it exists. PMID- 2971839 TI - Changes of molecular forms of atrial natriuretic peptide after treatment for congestive heart failure. AB - In the present study, an attempt was made to clarify whether ANP molecular forms in the plasma of severe congestive heart failure patients differ from those in healthy persons and whether ANP molecular forms in the plasma of the patients were changed by successful treatment of cardiac disease. Twenty patients with congestive heart failure were treated at Kitasato University Hospital. They were classified as class III or IV by New York Heart Association criteria at the time of admission. Plasma ANP concentrations decreased after treatment from 356 +/- 58.2 to 72.3 +/- 14.8 pg/ml. The gel permeation chromatograms from the plasma of healthy persons showed low, or low and high molecular weight ANP peaks which correspond to the elution positions of authentic alpha-ANP or ribonuclease A (mol. wt., 13.7 kdalton). In patients with severe congestive heart failure at a severe stage, middle molecular weight ANP consisted with the elution position of authentic beta-ANP was particularly noted in addition of low and high molecular weight ANP peaks. This middle molecular weight peak disappears in most of cases by successful treatment. Molecular forms in the plasma obtained from the coronary sinus and the inferior or superior vena cava were essentially the same. These results indicate that the middle molecular weight ANP supposed as beta-ANP may particularly be secreted in severe congestive heart failure patients. PMID- 2971840 TI - Improved technique for the mixed lymphocyte response in the Chacma baboon. AB - Lymphocyte responsiveness in cells collected from baboons (Papio ursinus orientalis) has consistently been 1/10 of that obtained using human cells. Animals that vigorously rejected organ allografts demonstrated that this lack of responsiveness was not due to genetic identity. This paper defines the kinetics of the baboon mixed lymphocyte response (MLR) and suggests several modifications to improve the sensitivity of the technique. Optimal response was obtained using nylon wool purified T-cell responders, 20% fresh autologous plasma to supplement culture medium, an incubation period of 6 days and exposure to 3H-thymidine (2 Ci/mmol) for 24 h prior to harvesting. These modifications enhanced the MLR response from a stimulation index +/- standard error of 2.5 +/- 0.3 (n = 79) to 25.2 +/- 5.4 (n = 38). PMID- 2971841 TI - Pharmacokinetics and metabolism of medroxyprogesterone acetate in patients with advanced breast cancer. AB - The pharmacokinetics and metabolism of medroxyprogesterone acetate (MPA) were studied in patients with advanced breast cancer after i.v. injection and oral administration of [3H]MPA. MPA was distributed very rapidly into three compartments after i.v. injections, revealing half-lives of 4-7 h. Using a nonlinear model fitting metabolic clearance rates (MCR) were found to be 652 1/day before and 601 1/day during MPA treatment, and distribution volumes (V0) 5.9 and 3.41 respectively. The major metabolite of MPA following i.v. injection was a glucuronide of MPA, presumably of the 3-enol form. After oral administration the radioactivity in serum increased rapidly and reached a plateau of about 1% of the dose per litre serum after approx 2 h. About 80-90% of the radioactivity was found in the water phase after hexane extraction, persumably as glucuronides of metabolites more polar than MPA. Radioimmunoassay (RIA) of MPA in untreated serum samples showed 3-8-fold greater MPA values as compared to measurements in hexane extracts of serum. Ethanol extraction did not remove these interfering substances. Extraction of serum with a low polar solvent before RIA of MPA is essential in order to prevent great overestimation, as the glucuronidated polar metabolites most likely will crossreact in an assay with an antiserum raised against a MPA-3-O-carboximethyloxime coupled to bovine serum albumin. PMID- 2971842 TI - Myeloid antigen expression on common acute lymphatic leukaemia blasts after culture. AB - Blast cells from seven out of ten patients with common acute lymphoblastic leukaemia (cALL) developed the myeloid antigen MY7 (CD13) after culture, and one of these coexpressed the myeloid antigen MY9 (CD33). CD13 expression appeared to be independent of maturation since it could be induced more readily in cultures which did not contain the differentiation promoter 12-O-tetradecanoyl-phorbol 13 acetate (TPA). CD13 expression in culture was not seen on one null ALL, or 6 B CLL investigated or on normal tonsillar B cells or PBMC under similar conditions. CD13 expression on cALL blasts probably represents evidence of abnormal gene expression in the leukaemic cells. However the absence of CD13 expression on the earlier B null ALL or the later B-CLL suggests we cannot exclude the possibility that CD13 expression is a feature of normal precursor B cells. PMID- 2971843 TI - Aging decreases beta-endorphin enhancement of T-cell mitogenesis in mice. AB - Numerous studies have demonstrated the role of the central nervous system in immunomodulation. beta-Endorphin, a neuropeptide that is released along with adrenocorticotropin by the pituitary in response to stress, has been shown to have various effects on immune function, although these effects are dependent on dose, animal model, and immune cell tested. Since the increased risk for infection and tumor that is observed in the elderly is thought to be in part secondary to waning cell-mediated immunity, we investigated the effect of age on beta-endorphin immunomodulation of T-cell proliferation in a murine model. Spleen cells obtained from young and old BALB/c mice were cultured in vitro with various mitogens with and without beta-endorphin. beta-Endorphin at 10(-8) M on day 3 of culture significantly enhanced concanavalin A (2.0 micrograms/10(6) cells per ml) mitogenesis but not phytohemagglutinin or lipopolysaccharide mitogenesis. Moreover, this enhancement was shown only in spleen cells from young mice and was not blocked by the opiate receptor antagonist naloxone, which suggests that enhancement of mitogenesis by beta-endorphin was mediated by a non-opiate receptor. Finally, our results support an altered response to neuroimmunomodulation with age. PMID- 2971844 TI - Analysis of immunocompetent cells following inner ear immunostimulation. AB - Immunohistochemical methods were used to investigate the development of inner ear immunocompetent cells in healthy, BALB/c mice over a 3-week period as a result of a secondary inner ear response against keyhole limpet hemocyanin. Antibodies against murine macrophages and granulocytes (anti-Mac-1), T-cells (anti-Lyt-1, anti-Lyt-2), and immunoglobulins (anti-IgM, anti-IgG, anti-IgA) were used. Mac-1 positive (Mac-1+) cells were observed at 6 hours post-challenge in the endolymphatic sac and cochlea and rapidly increased in both sites. Lyt-1+ cells gradually increased in the endolymphatic sac after challenge, peaking at 2 and 3 weeks post-challenge. In the cochlea, Lyt-1+ cells were detected at 1 day post challenge and then increased. Lyt-2+ cells were detectable in the endolymphatic sac and the cochlea by 3 weeks post-challenge. The predominant immunoglobulin bearing cell in the endolymphatic sac was IgG, followed by IgM, with IgA seen late in the response. We conclude that the inner ear has the capacity to mount an immune response through the accumulation of the needed immunocompetent cells for antigen processing, antibody production, and modulation of the response through T helper and suppressor cell activity. PMID- 2971845 TI - Laser balloon angioplasty: effect of exposure duration on shear strength of welded layers of postmortem human aorta. AB - Laser Balloon Angioplasty (LBA) is a technique that may improve the results of balloon angioplasty by thermally sealing arterial dissections and reducing elastic recoil. To define the relationship between laser-exposure duration and the strength of thermal welds made between separated layers of arterial wall, 360 1-cm discs of human postmortem aorta were lased for six different exposure intervals at three different temperature ranges, comparing shear strength of thermal welds in the different groups. Twenty discs were lased to achieve plateau adventitial temperatures of 95 degrees C-104 degrees C (group A), 105 degrees C 114 degrees C (group B), or 115 degrees C-124 degrees C (group C) at each of the exposure periods (5, 10, 15, 20, 25, and 30 sec). A 400-micron fiberoptic coupled to a 1.06 micron continuous wave neodymium:YAG laser was placed perpendicularly 8 mm above the luminal surface of each disc, which had been split midway between the intimal and adventitial surface and reapposed. Mean laser energy ranged 78 378 J delivered in a decremental stepwise fashion to achieve quickly and maintain the target plateau tissue temperature. Mean weld strength increased in relation to both achieved tissue temperature and laser-exposure duration, with at least 10 sec necessary, at temperatures greater than 95 degrees C, for reliable thermal welding. Laser exposure for greater than 20 sec provided no statistical increment in weld strength. In the anticipated clinical performance of LBA, these data suggest that when thermal fusion of disrupted arterial tissues is desired, a laser-exposure duration of 10-20 sec is optimal. PMID- 2971846 TI - Management of rhinophyma with carbon dioxide laser: Lahey Clinic experience. AB - The carbon dioxide (CO2) laser as a surgical tool for the difficult cosmetic problem of rhinophyma permits unprecedented refinement in treatment. Previously described conventional techniques have included the cold knife and the dermabrader. With these methods, hemostasis must be obtained with electrocautery, which if used extensively makes fine sculpting difficult. Seven patients with rhinophyma were treated with the CO2 laser at the Lahey Clinic from 1982 to 1987. We used a laser handpiece with a variable spot size. With this technique, we found the laser to be an excellent cutting tool while simultaneously providing superior hemostasis. In the followup period of up to 4.5 years, the cosmetic results have been excellent. Thus, the CO2 laser has become our treatment of choice for the management of patients with rhinophyma. PMID- 2971847 TI - Combined effects of hematoporphyrin derivative phototherapy and adriamycin in a murine tumor model. AB - Hematoporphyrin derivative phototherapy (HpD-PT) is currently being used experimentally in the treatment of various malignancies. The effectiveness of HpD PT appears to be limited to superficial malignancies because of its inability to penetrate more than 0.5-1 cm. The mechanism of action has been demonstrated to result from the formation of toxic singlet oxygen and other oxygen radicals. Preliminary work suggested that the addition of other chemical agents known to produce toxic oxygen radicals may enhance the effect of HpD-PT. To test this hypothesis, we examined the effectiveness of HpD-PT in the Walker 256 carcinosarcoma with and without Adriamycin. Tumor kill was estimated by calculating the area of necrosis on photomicrographic sections. From this area, a radius of tumor necrosis was determined and compared among various control and experimental groups. HpD-PT was effective in this model when compared to control groups (radius of necrosis 3.79 +/- 0.97 mm vs. 2.60 +/- 1.26 mm, P less than .05). The addition of Adriamycin significantly increased the radius of necrosis when compared to HpD-PT (4.57 +/- 1.70 mm vs. 3.69 +/- 0.97 mm, P less than .01). From this work we conclude that HpD-PT demonstrates effective tumor killing in this murine model. The addition of Adriamycin significantly increased tumor kill. PMID- 2971848 TI - Neuronal damage in the albino rat: excitotoxin, microtubule inhibitor synergism. AB - Separate lines of investigation implicate excitotoxins or disruption of cytoskeletal architecture in the pathogenesis of human neurodegenerative disease. To assess the hypothesis that these neurotoxic mechanisms may have a synergistic effect, albino rats were given intrastriatal administrations of either the excitotoxin, kainic acid, or the microtubule inhibitor, colchicine, or both. Assessment of neuronal damage, either by loss of neurotransmitter metabolic enzyme activities or by histologic analysis, demonstrated synergism between the agents. Further, the synergism was still apparent with substitution of quinolinic acid, but not dimethyl kainate, for kainic acid, and Vinca alkaloids, but not lumicolchicine, for colchicine. PMID- 2971849 TI - Plasma and atrial levels of atrial natriuretic peptide (ANP) in pulmonary hypertensive rats. AB - Immunoreactive atrial natriuretic peptide (IR-ANP) was measured in plasma and atrium of normal and monocrotaline induced pulmonary hypertensive rats (PH rats). In these animals, there was right ventricular hypertrophy and right ventricular systolic pressure was elevated. Fourteen days after a single dose of monocrotaline (40 mg/kg), plasma IR-ANP concentrations were significantly elevated (964.3 +/- 63.0 pg/ml vs. 521.0 +/- 81.9 pg/ml in controls, p less than 0.001). Tissue levels of IR-ANP in the right atrium in PH rats was significantly lower than those in the controls (45.1 +/- 3.9 ng/mg vs. 240.5 +/- 10.4 ng/mg, p less than 0.001), while there was no significant difference in tissue levels of atrial IR-ANP in the left atrium between the two groups. Thus, development of pulmonary hypertension led to an increase in release of ANP from the right atrium. PMID- 2971850 TI - Time-dependent enhancement of lymphocyte activation by mitogens after exposure to isolation or water scheduling. AB - The effects of isolation and water scheduling on mitogen induced lymphocyte proliferation were investigated. Isolated rats were animals which had been raised in group-housed conditions and then transferred to individual cages with ad lib access to water for a 1 or 2 week period. Water scheduled rats were maintained in group housing (5 rats per cage) with ad lib access to food but with access to water for a single 30 minute session each day. Responses of these groups were compared to those of animals which had been continuously group-housed with ad lib access to food and water. No differences in lymphocyte responses to phytohemagglutinin (PHA) were found 1 week after exposure to isolation. However, after 2 weeks, splenic and blood T lymphocytes from isolated animals demonstrated an increased proliferative response to suboptimum and maximum concentrations of PHA. Splenic B lymphocyte responses to lipopolysaccharide (LPS) from isolated animals were also increased by 2- to 3-fold compared to group-housed controls. Two weeks of exposure of animals to daily water scheduling similarly increased the splenic lymphocyte proliferation. This increased responsiveness to PHA was not accompanied by a significant change in the sensitivity of the lymphocytes to PHA, in the total number of white blood cells, or the proportion of splenic T or T helper lymphocytes. Our results show that the increase in lymphocyte proliferation is time-dependent, requires greater than 1 week of exposure to isolation and is due to factors other than changes in sensitivity to mitogen or T lymphocyte number. PMID- 2971851 TI - h-ANF does not play a role in the regulation of myocardial force of contraction. AB - In the present study, h-ANF failed to reduce myocardial force of contraction alone and in the presence of isoprenaline. This lack of effect was observed in electrically driven, isolated human papillary muscle strips and rat papillary muscles. In contrast, adenosine and carbachol exerted antiadrenergic actions in rat and human myocardium. It is concluded that h-ANF obviously does not play a role in the regulation of human and rat myocardial force of contraction, whereas adenosine and carbachol are capable to produce antiadrenergic effects in the human and rat myocardium. PMID- 2971852 TI - Treating endometriosis with nafarelin. PMID- 2971853 TI - [Lithium and psoriasis. Critical review and case reports]. PMID- 2971854 TI - Postoperative monitoring of microsurgical free tissue transfers for head and neck reconstruction. AB - Approximately 10% of free tissue transfers fail because of thrombosis of the arterial or venous anastomoses. Postoperative monitoring to allow early detection of impaired flap perfusion is crucial, especially in head and neck reconstruction, in which failure of a free flap may be associated with significant morbidity. Furthermore, the increasing use of "buried" free tissue transfers in the head and neck precludes direct clinical observation. The various techniques that have been advocated for clinical monitoring of free tissue transfers are reviewed, and their potential applications in head and neck reconstruction are discussed with specific emphasis on the use of low-frequency continuous-wave Doppler surface monitoring and implantable high-frequency pulsed ultrasonic Doppler monitoring. PMID- 2971855 TI - Splanchnic disposal of human atrial natriuretic peptide in humans. AB - In healthy men (n = 6), the splanchnic fractional extraction of human atrial natriuretic peptide (hANP), as determined by the hepatic venous catheter technique, was 75% under basal conditions resulting in a splanchnic uptake of hANP of 8.5 +/- 5.0 pmol/min. In spite of a drop (P less than .05) in splanchnic fractional extraction to about 50%, splanchnic uptake of hANP rose to 56 to 99 pmol/min (P less than .01) when pharmacologic plasma concentrations of hANP were induced during a bolus (100 micrograms)-primed intravenous (IV) infusion (100 micrograms/h; time, one hour) of hANP. This was accompanied by a fall in estimated hepatic blood flow (P less than .05), in pulmonary arterial pressure (P less than .01), and, in each individual, in systemic BP. Total metabolic clearance rates, splanchnic clearance rates, and production rates of hANP were 4.5 +/- 2.2 L/min, 0.4 +/- 0.1 L/min, and 46.1 +/- 20.1 pmol/min, respectively. Thus, in healthy men, the splanchnic area accounts for approximately 10% of total hANP clearance. PMID- 2971856 TI - Enhanced antimicrobial activity of lactoferrin by binding to the bacterial surface. AB - In order to elucidate the mechanisms involved in the antibacterial activity of lactoferrin, quantitative determinations of siderophore production and lactoferrin adsorption on various bacterial species were performed. The binding of lactoferrin took place both on Gram-positive and Gram-negative species and occurred with bacterial cells grown in stress or in excess of iron. The different degrees of sensitivity to lactoferrin observed could not be directly related to the type and amount of siderophores produced. However, it was possible to find a correlation between the capacity of some strains to bind lactoferrin and their sensitivity to this protein. These data suggest that the binding of lactoferrin on the cell surface can result in an antibacterial activity additional to iron witholding. PMID- 2971857 TI - Possible regulation of autologous mixed lymphocyte reaction by OKM1+ NK cells. AB - T cells are stimulated by autologous non-T cells and interleukin 2 (IL-2) is produced in the conventional autologous mixed lymphocyte reaction (AMLR) in young healthy controls. The role of cells with natural killer (NK) cell markers (OKM1+ cells or Leu 7+ cells) in the AMLR was studied. There were significant inverse correlations between the percentage of input OKM1+ cells minus monocyte (OKM1+ NK cells) and either AMLR proliferation (gamma = -0.9, P less than 0.001) or IL-2 production (gamma = -0.75, P less than 0.01) in the AMLR cultures after 7 days measured at 7 days. A statistically significant correlation was observed between the percentage of input Leu 7+ cells and AMLR proliferation (gamma = -0.64, P less than 0.05), but not IL-2 production. These results suggest that the AMLR is controlled by OKM1+ NK, perhaps acting through IL-2 regulation. PMID- 2971858 TI - HIV virus, prostaglandins and essential fatty acids; a suggested mechanism for T helper cell penetration. AB - Prostaglandins and or its precursors are present in certain body fluids. It has been suggested that critical concentrations of PGE are essential for the HIV to penetrate a lymphocyte. Certain clinical implications of this hypothesis have been discussed. PMID- 2971859 TI - Neurological complications of total heart replacement: a proposed role for atrial natriuretic factor. PMID- 2971860 TI - Precision surgery with an electromagnetically induced current convergence probe application in aneurysm treatment, angioplasty, and brain tumor resection in in vivo and in vitro models. AB - A hand-held probe, or one introduced through a catheter, rapidly produces an extremely high, tissue-vaporizing temperature in a precisely defined manner enabling surgeons or interventional radiologists to perform angioplasty, thrombose aneurysms, and vaporize tumors. The probe is operated in a near field of an inductive coil, and the current induced in the biologic tissue is converged maximally at the tip of the probe at the resonance frequency of both the inductor and the probe, producing a maximum temperature in excess of 1400 degrees C. Radio frequency power controls the probe-tip temperature. The operation of the probe is comparable to that of a CO2 or YAG laser and is complementary to laser-surgical techniques. The low cost relative to lasers and simplicity of the device including its disposable components make the prospect of commercialization of this device promising. PMID- 2971861 TI - Acne fulminans with dapsone induced haemolysis: a case report. PMID- 2971862 TI - [Observations on a case load of workers occupationally exposed to corticosteroids]. PMID- 2971863 TI - [Epidemiological surveillance of natural foci of tick-borne encephalitis in the recreational area of a large city (the methodology and results]. PMID- 2971864 TI - [Isoenzyme analysis of the Leishmania major strains isolated from patients with zoonotic cutaneous leishmaniasis in different sections of the endemic territory in the USSR]. PMID- 2971865 TI - [Biochemical structure and allergenic activity of dermatophagoid mites (Dermatophagoides pteronyssinus)]. PMID- 2971868 TI - [Down's syndrome with unusual karyotype: tandem duplication of chromosome 21]. PMID- 2971866 TI - [Cilazapril in essential hypertension. A study of one year of continuous treatment]. PMID- 2971867 TI - Cycloheximide blocks nicotinamide action on phosphate excretion in thyroparathyroidectomized rats. AB - Nicotinamide causes marked inhibition of phosphate (Pi) reabsorption in the renal proximal tubule and this can be detected both as a specific decrease in Na+ dependent Pi uptake by isolated brush border membrane (BBM) vesicles and as a marked increase in urinary Pi excretion. In the present study, thyroparathyroidectomized (TPTX) rats were adapted to low-Pi diet and injected with cycloheximide (CHX), an inhibitor of protein synthesis, prior to nicotinamide treatment. Compared to rats that received nicotinamide alone, the pretreatment with CHX did not alter the inhibitory action of nicotinamide on BBM uptake of Pi. However, CHX completely blocked the phosphaturic response. We conclude that, at least in TPTX rats adapted to low-Pi diet, nicotinamide inhibits Pi reabsorption not only in the proximal convoluted tubule but also in other nephron segments such as the pars recta or distal sites. Nicotinamide action on these additional sites is dependent on protein synthesis, in contrast to its action in the proximal convoluted tubule. PMID- 2971869 TI - [Thyroid and adrenal response to acute stimulation with HCG: effects of bilateral ovariectomy]. PMID- 2971870 TI - Multistate outbreak of sporotrichosis in seedling handlers, 1988. PMID- 2971871 TI - Thymidylate synthase as a determinant of 5-fluoro-2'-deoxyuridine response in human colonic tumor cell lines. AB - A panel of seven human colorectal cell lines of differing phenotype has been examined to elucidate the role of thymidylate synthase (TS) in the response to 5 fluoro-2'-deoxyuridine (FdUrd). Although TS is a major target of FdUrd, no consistent relationship was observed between the intracellular levels of TS and the response to FdUrd among the cell lines. Levels of thymidine kinase and dihydrofolate reductase, enzymes that are involved in generation of ligands that form the inhibitory ternary complex with TS, do not correlate with FdUrd response. Two cell lines that exhibit innate resistance to FdUrd, relative to the other cell lines, have variations in TS enzyme structure or gene structure. Cell line HCT 116 contains two forms of TS, as defined by isoelectric focusing. One form, which is unique to HCT 116, is more basic than the common form, which is present in all the cell lines. Cell line RCA contains a variation in the TS structural gene, as defined by restriction fragment-length analysis. These structural variations, which are associated with reduced response to FdUrd, may serve as markers for reduced clinical response to TS-directed chemotherapy. PMID- 2971872 TI - A naturally occurring variation in thymidylate synthase structure is associated with a reduced response to 5-fluoro-2'-deoxyuridine in a human colon tumor cell line. AB - Inhibition of thymidylate synthase (TS) is an important mechanism of action of fluoropyrimidine antimetabolites. Thus, TS structure and expression are expected to be determinants of response to these agents. The role of TS in fluoropyrimidine response has been analyzed in a panel of human colonic tumor cell lines. Previous work has demonstrated that there is little correlation between TS concentration and sensitivity to 5-fluoro-2'-deoxyuridine (FdUrd) among these cell lines, suggesting that parameters other than the TS levels are responsible for the variations in drug response. One such parameter has been identified in cell line HCT 116. This line, which is relatively resistant to FdUrd, produces two structural forms of TS, as determined by mobility of the enzyme in isoelectric focusing polyacrylamide gels. One form is common to all the cell lines, whereas a variant form, which is more basic and is encoded by a separate structural gene, is unique to HCT 116. Cells expressing one or the other TS form have been isolated and used to demonstrate that the variant form is associated with FdUrd resistance. Kinetic experiments indicate that the variant TS has reduced affinities for 5-fluoro-2'-deoxyuridylate and 5,10 methylenetetrahydrofolate, which are ligands involved in formation of a stable inhibitory complex with the enzyme. Thus, the innate resistance of cell line HCT 116 to FdUrd is derived, at least in part, from production of an altered structural form of TS having reduced affinity for ligands. PMID- 2971874 TI - Effect of exercise on glycolytic enzymes of Zucker fatty rats. AB - The effect of fatigue (running to exhaustion) on the Vmax activity of the key glycolytic enzymes measured at saturating substrate concentrations in muscles, liver and brain of sedentary and trained (running on a treadmill one h/day at 20 m/min, five days/week for six months) female Zucker fatty rats and their lean littermates was investigated. In the sedentary rats, fatigue increased the activity of phosphofructokinase (PFK) in the red vastus muscle by 82% in lean, and 120% in obese rats. In the trained rats, fatigue increased PFK activity by 28% in the white vastus muscle of lean rats. In the lean animals, hexokinase (HK) activity was decreased by 26% in the red vastus of sedentary rats, and by 29% in the white vastus of trained rats upon fatiguing. Pyruvate kinase (PK) activity was also decreased by 29% in the white vastus of fatigued lean animals. Training by itself had no effect on the activity of glycolytic enzymes, except PK activity which was increased by 27% in the cortex of the lean animals. It is concluded that in the Zucker rat, these glycolytic enzymes may play a differential role in regulating glycolysis during exercise and fatigue; the extent of their involvement differs depending upon the type of tissue studied and exercise. In view of the reported short half-life (7-17 h) of PFK and its covalent modification, it is suggested that the total content and/or phosphorylation status of the enzyme may be affected in animals subjected to long-term fatigue. PMID- 2971873 TI - Purification and characterization of a Ca2+/Mg2+ ecto-ATPase from rat heart sarcolemma. AB - The Ca2+/Mg2+ ATPase of the rat heart sarcolemmal particles was solubilized with Triton X-100 after treating the membranes with trypsin and purified by high speed centrifugation, ammonium sulfate fractionation, hydrophobic chromatography and gel filtration. The purified enzyme was seen as a single protein band in non denaturing polyacrylamide gel electrophoresis and its molecular weight by gel filtration was found to be about 240,000. The enzyme utilized Ca-ATP or Mg-ATP as a substrate with high affinity sites (Km = 0.12-0.16 mM) and low affinity sites (Km = 1 mM). The enzyme also utilized CTP, GTP, ITP, UTP and ADP as substrates but at a lower rate in comparison to ATP. The enzyme was activated by Ca2+ (Ka = 0.4 mM) and Mg2+ (Ka = 0.2 mM) as well as by other cations in the order Ca2+ greater than Mg2+ greater than Mn2+ greater than Sr2+ greater than Ba2+ greater than Ni2+ greater than Cu2+. The ATPase activity in the presence of Ca2+ was markedly inhibited by Mg2+, Mn2+, Ni2+ and Cu2+ whereas the monovalent cations such as Na+ and K+ were without effect. The enzyme did not exhibit Ca2+ stimulated Mg2+ dependent ATPase activity and was insensitive to calmodulin, ouabain, verapamil, D-600, oligomycin, azide and vanadate. Optimum pH for Ca2+ or Mg2+ ATPase activity was 8.5-9.0. In view of the possible ectoenzyme nature of the ATPase, its role in adenine nucleotide and Ca2+ metabolism in the myocardium is discussed. PMID- 2971875 TI - The binding of IgG1 containing immune complexes to the FcR of allogenically activated T cells induces changes in the membrane potential and the cell surface charge. AB - The effect on membrane potential and cell surface charge of binding immune complexes containing IgG1 and IgG2a monoclonal antibodies to Fc receptors was studied in resting and allogenically activated murine T cells. IgG1 complexed by antigen or heat aggregation induced electrophysiological changes on activated T cells. A biphasic alteration of membrane potential was detected by measurement of the intra- and extracellular distribution of the fluorescent dye, DiOC6. A short lived hyperpolarization, detectable for 4-6 min after adding the respective ligand, was followed by a longer lasting depolarization. The cell surface charge, measured by cell electrophoresis, was also changed. This alteration was detected 2-4 hr after addition of immune complexes and disappeared by the 8th hr of incubation. Monoclonal antibody 2.4.G2, reactive with mouse FcR, induced a similar membrane potential response on activated T cells, but did not affect the cell surface charge. Monomeric IgGs and complexes of IgG2a did not modify these parameters. FcR ligands had no effect on the studied characteristics of resting T cells. PMID- 2971877 TI - Suppressor T cells in myasthenia gravis. PMID- 2971876 TI - Non-receptor striated muscle antibodies in sera from patients with myasthenia gravis. PMID- 2971878 TI - [Ultrasonographic morphology of liver diseases in the light of laparoscopy]. PMID- 2971879 TI - Myosin ATPase intermediate density fibers for diagnosis of reinnervation. AB - Myosin ATPase (pH 9.4) differentiates two muscle fiber types in healthy human muscle, while diseased muscle often contains intermediate density fibers (IDFs). We evaluated the possibility that, since almost all IDFs in pathologic muscle biopsies are changing type after reinnervation by a motor axon of the opposite type, IDFs are useful in diagnosis. In a retrospective study of 208 muscle biopsies, IDFs were seen as often as were esterase-positive angular atrophic fibers (EPAAFs). In denervation identified by EMG and by histopathology, EPAAFs and IDFs were found much more often than were other indicators. Of biopsies diagnosed without use of IDFs as minimal histologic change or no pathologic diagnosis, 16% had IDFs with sparse EPAAFs and 21% had IDFs without EPAAFs, suggesting mild denervation with rapid reinnervation. IDFs correlate well with EPAAFs, identifying reinnervated versus denervated fibers. Type grouping reveals completed reinnervation change that may be many years old, while IDFs are changing type when biopsied and thus reveal recent reinnervation and preceding denervation. IDFs usefully belong with the histochemical indicators used to evaluate muscle disease. PMID- 2971880 TI - Impaired response of atrial natriuretic factor to high salt intake in persons prone to hypertension. PMID- 2971881 TI - Novel anti-inflammatory peptides from the region of highest similarity between uteroglobin and lipocortin I. AB - Significant future developments in the effective treatment of inflammatory diseases may arise from non-toxic dual inhibitors of both cyclooxygenase and lipoxygenase pathways in the arachidonate cascade. Inhibition of phospholipase A2(PLA2)(EC3.1.1.4), may provide such a dual action and recent research has concentrated on the role of PLA2-inhibitory proteins as possible anti inflammatory agents. Blastokinin or uteroglobin is a steroid-induced rabbit secretory protein with PLA2-inhibitory activity. Its biochemical and biological properties have been extensively studied and its crystallographic structure has been resolved at 1.34 A (refs 15, 16). Lipocortins are a family of related proteins, which, it has been suggested, mediate the anti-inflammatory effects of glucocorticoids (for a review, see ref. 23). Some proteins of this group have been purified and the complementary DNA sequences of two human lipocortins are known. Lipocortins inhibit PLA2 in vitro, although their mechanism of action is still unclear. Recombinant lipocortin I inhibits eicosanoid synthesis in isolated perfused lungs from the guinea pig. Here, we report that synthetic oligopeptides corresponding to a region of high amino-acid sequence similarity between uteroglobin and lipocortin I have potent PLA2 inhibitory activity in vitro and striking anti-inflammatory effects in vivo. PMID- 2971882 TI - [Down's syndrome and dementia: a growing problem in the care of the mentally retarded]. PMID- 2971883 TI - [The relation between Down's syndrome and Alzheimer-type dementia]. PMID- 2971884 TI - [Corpus luteum hemorrhage; beware of overtreatment]. PMID- 2971886 TI - [The reversal of the medical burden of proof]. PMID- 2971885 TI - [Injuries of the oral mucous membrane]. PMID- 2971887 TI - [Sterilization of handicapped within the scope of a discussion outline for a new guardianship law]. PMID- 2971888 TI - Atrial natriuretic factor. What's in a name? PMID- 2971889 TI - A novel effect of molybdate on the binding of [3H]aldosterone to gel-filtered type I receptors in brain cytosol. AB - Recently we reported that adding molybdate to crude steroid-free cytosol at 0 degree C results in a dose-dependent reduction in the binding of [3H]aldosterone ([3H]ALDO) to Type I adrenocorticosteroid receptors. In the experiments outlined here, we found that addition of molybdate to steroid-free brain cytosol produces a 30-50% increase in the subsequently measured maximal specific binding capacity (BMAX) of [3H]ALDO-Type I receptors if the cytosol is subjected to Sephadex G-25 gel filtration prior to steroid addition. These manipulations were found to have no effect on the equilibrium dissociation constant (Kd) of the receptors. In contrast, when gel filtration of steroid-free cytosol was performed in the absence of molybdate, there was a 2-fold increase in the Kd and over a 50% reduction in the subsequently measured BMAX of [3H]ALDO-Type I receptors. When molybdate was added to this steroid-free cytosol immediately following gel filtration, there was no reduction (or increase) in Type I receptor [3H]ALDO binding capacity compared with non-gel-filtered controls. The addition of as little as 2 mM molybdate to crude steroid-free cytosol was found to stabilize the binding capacity of Type I receptors during exposure to 22 degrees C incubations; however, when gel-filtered steroid-free cytosol was exposed to these conditions at least 10 mM molybdate was required to stabilize Type I receptor binding capacity. Adding the sulfhydryl reducing reagent, dithiothreitol, to the various steroid-free cytosols had little effect on [3H]ALDO-Type I receptor binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971890 TI - Multilevel lumbar laminotomies for focal spinal stenosis: case report. AB - We describe a case of spondylotic lumbar spinal stenosis in an infirm elderly patient successfully treated with multilevel laminotomies. This procedure is less stressful to patients than a full laminectomy. In a selected subgroup of patients with "hourglass" spondylotic stenosis, it may provide adequate decompression. PMID- 2971891 TI - Increased concentration of atrial natriuretic factor in the cerebrospinal fluid of patients with aneurysmal subarachnoid hemorrhage and raised intracranial pressure. AB - Plasma and cerebrospinal fluid (CSF) atrial natriuretic factors/peptides (ANFs/ANPs) were measured in 26 patients with normal or raised intracranial pressure (ICP) by means of an instant radioreceptor assay. All 26 patients were suffering from aneurysmal subarachnoid hemorrhage (SAH), and 11 had also developed raised ICP (ICP greater than 20 mm Hg). In SAH patients with normal ICP, the plasma levels of ANF were 20 to 200 pg/ml (mean +/- SE, 89 +/- 68 pg/ml); in the 11 SAH patients with raised ICP, however, ANF levels were 14 to 262 pg/ml (mean 114 +/- 79 pg/ml). The difference was not statistically significant. The ANF/ANP plasma levels in 6 healthy volunteers were 15 to 167 pg/ml (mean 77 +/- 32 pg/ml). Although the ANF/ANP concentration in the CSF of patients with normal ICP did not reach the lower limit of detectability (i.e., 4 pg/ml) in any case, in those with elevated ICP it was 14 to 120 pg/ml (mean 49 +/ 37 pg/ml). This difference was statistically highly significant. The results of this preliminary study suggest that the ANF/ANP concentration in human CSF is 1 to 2 orders lower than that in the plasma and that there is no significant correlation between ANF/ANP levels in the CSF and the plasma. After SAH in patients with raised ICP, there was an accompanying increase in the ANF/ANP concentration in the CSF, but the ANF/ANP concentration in the plasma was not changed significantly. Accordingly, a central ANF/ANP release might be hypothesized to play a causative or adaptive role in the neuroendocrine regulation of ICP dynamics, although this may simply be an epiphenomenon. PMID- 2971892 TI - Sparing of cholinergic neurons following quinolinic acid lesions of the rat striatum. AB - A combination of immunocytochemical and enzyme histochemical methods have been used to study those neurons which survive lesions of the rat striatum, produced by low doses of the excitotoxin quinolinic acid. Nissl-stained sections revealed that following injection of this toxin many large neurons remained within areas of extensive cell loss. These large cells were found to express both the enzyme acetylcholinesterase and choline acetyltransferase-like immunoreactivity. The surviving cells did not contain the enzyme reduced nicotinamide adenine dinucleotide phosphate or the peptides, somatostatin and neuropeptide Y. This pattern of selective cell sparing was also found following lesions induced by low doses of the toxins ibotenic acid and kainic acid. The survival of large neurons indicates that the excitotoxin-lesioned rat striatum shares common features with the pattern of cell loss found in the caudate-putamen in Huntington's disease. The major difference between these two examples of striatal nerve cell degeneration is, however, the selective preservation of somatostatin/neuropeptide Y/nicotinamide adenine dinucleotide phosphate-diaphorase-containing neurons found in Huntington's disease but not observed following quinolinic acid lesions. PMID- 2971894 TI - [Spigelian hernia: diagnosis and therapy]. PMID- 2971893 TI - [Electrocardiography in chronic alcoholics]. PMID- 2971896 TI - [Redistribution of intrahepatic arterial blood flow after ligation of an aberrant lobar artery. Use of degradable starch microspheres]. PMID- 2971895 TI - [Possibilities and limitations of a myocutaneous rectus abdominis flap in the reconstruction of the breast after mastectomy]. PMID- 2971897 TI - [A case of ovarian dysgerminoma in Down's syndrome]. PMID- 2971898 TI - Non-operative revascularisation of renal artery occlusion by transluminal angioplasty. AB - A modified transluminal angioplasty technique for treatment of renal artery occlusion has been developed. From 1980 to 1985, 16 consecutive patients with 17 complete main renal artery occlusions underwent interventional transfemoral angiography for the purpose of recanalisation. In 12 patients the orifice of the renal artery could be clearly localised, so an attempt was made. Successful revascularisation of the occluded vessel was accomplished in seven patients. In four of these seven, contralateral renal artery stenosis was detected and dilated at the same session. Intact vasculature could be demonstrated distal to the occlusion. In six patients an improvement of renal function was apparent at the end of the dilatation procedure; this was indicated by the appearance of contrast material in the pelvicalyceal system. The mean serum creatinine fell from 4.6 +/- 2.9 to 1.9 +/- 0.4 mg/dl. Radioisotope studies confirmed improvement of renal function in the previously occluded kidney in four of four patients. Three patients had acute oliguric renal failure, which was reversible in two cases following revascularisation. Transluminal angioplasty improved hypertension in all cases. Mean blood pressure fell from 187/110 to 155/88 mmHg. Non-operative renal artery revascularisation can be achieved by transluminal angioplasty techniques and is an alternative to surgery in patients with increased operative risk. PMID- 2971900 TI - Percutaneous transluminal angioplasty of renal transplant artery stenosis in patients with rejection nephropathy. AB - A total of 13 procedures of percutaneous transluminal angioplasty were performed in 11 kidney graft recipients with renal transplant artery stenosis. Nine procedures were technically successful in eight patients (one redilatation was necessary because of restenosis). Graft biopsy confirmed rejection nephropathy in all cases. The outcome could not be evaluated in one patient who died of a concomitant disease shortly after angioplasty. The remaining seven patients (eight percutaneous transluminal angioplasties) showed improvement in hypertension in 63% and in glomerular filtration rate in 50% of procedures, persisting for 6-13 months. A single major complication encountered was a loss of graft related to unsuccessful percutaneous transluminal angioplasty. The results suggest that percutaneous transluminal angioplasty may bring prolonged increase in effective renal plasma flow and glomerular filtration rate even in patients experiencing rejection nephropathy. PMID- 2971899 TI - Presence of the C3b complement receptor (CR1) antigen in the glomerular basement membrane of human fetal kidneys. AB - The distribution pattern of the C3b receptor (CR1) antigen, one of the earliest differentiation markers of the podocyte plasma membrane, has been studied during human renal ontogenesis by indirect immunofluorescence and immunoelectronmicroscopy using Fab'2 fragments of anti-CR1 antibodies. CR1 antigen has been detected at the open-stage glomerulus, which precedes constriction of the vascular pole, along the capillary wall, in a thick and homogeneous pattern of staining. By immunoelectronmicroscopy, CR1 antigen was present on the porocyte plasma membrane towards the glomerular basement membrane (GBM), and diffusely distributed within the GBM according to a decreasing intensity gradient from podocytes to endothelial cells. At a later stage of differentiation ('closed' glomerulus stage) when constriction of the vascular pole has occurred, the podocyte plasma membrane appeared diffusely labelled for CR1; the GBM was also stained with a decreasing intensity gradient from podocytes to endothelial cells. This result indicates that a plasma membrane receptor belonging to the cell coat of podocytes can be found in the extracellular space within the GBM. Thus the antigenic repertoire of the GBM of human fetal kidney comprises antigens of constituent molecules of the extracellular matrix and planted antigens originating from adjacent cells. PMID- 2971902 TI - Industrial deafness. PMID- 2971901 TI - Inhibitory effect of brain natriuretic peptide on central angiotensin II stimulated pressor response in conscious rats. AB - The effect of intracerebroventricular injection of brain natriuretic peptide (BNP) on blood pressure and heart rate was studied in conscious, unrestrained rats. The intracerebroventricular (i.c.v.) injection of BNP did not affect basal blood pressure and heart rate, but it attenuated i.c.v. angiotensin II (AII) induced pressor response in a dose-dependent manner. These results suggest that BNP plays a role in the regulation of blood pressure in the central nervous system. PMID- 2971903 TI - Back injury. PMID- 2971904 TI - Excision of pelvic endometriosis with the carbon dioxide laser laparoscope. AB - Laparoscopic vaporization of peritoneal endometriosis has been reported to relieve pain and enhance fertility. However, vaporization of endometriosis, particularly large plaques and hemorrhagic areas, is associated with significant amounts of carbon deposition in the desicated tissues. Further, it is difficult to ascertain complete removal of disease in some areas. In this study, 66 patients were treated by excision of peritoneal and/or ovarian endometriosis with the carbon dioxide laser via laparoscopy. Using excisional techniques, we were able to visualize the extent of peritoneal penetration and subperitoneal spread of endometriotic nodules. Further, we obtained histologic confirmation of the disease process in all patients. No patient received postoperative hormonal therapy during follow-up. Follow-up evaluation of these patients revealed relief of the painful symptoms of endometriosis. No long-term complications were encountered. PMID- 2971905 TI - Minor trauma back pain. PMID- 2971906 TI - Hysteroscopic management of mullerian fusion defects. AB - Operative hysteroscopy for mullerian fusion defects in properly selected cases and in experienced hands is a safe and effective method of management of uterine septal defects associated with recurrent pregnancy losses. For the patient, it is a more acceptable alternative because it avoids laparotomy, a lengthy hospital stay, and blood transfusions. Medical costs are reduced compared with more traditional approaches. While providing the same level of success, this technique makes a vaginal delivery possible. PMID- 2971907 TI - Use of general anesthesia in dental care of the child with sickle cell anemia. A case report. AB - This article reviews the clinical manifestations of sickle cell anemia and describes the dental management, with the use of general anesthesia, of a child affected by the disease and exhibiting some of its complications. The discussion stresses the vital importance of awareness of the possible complications of sickle cell anemia and sickle cell trait in the safe and successful management of patients requiring dental care that includes the use of general anesthesia. PMID- 2971908 TI - Management of dental patients with bleeding disorders: review and update. AB - Management of the dental patient who has a bleeding disorder requires an understanding of the normal hemostatic system and the patient's specific coagulation defect. This patient group can receive quality comprehensive dental care, provided appropriate preoperative planning and evaluation with the patient's physician or hematologist is accomplished. Emphasis should be placed on providing appropriate replacement therapy before the dental procedure, selection of conservative treatment approaches, and use of local hemostatic measures to facilitate hemostasis. PMID- 2971909 TI - Management of sports injuries in children and adolescents. AB - In general, child and youth sports are safe. The vast multitude of injuries sustained are mild contusions, sprains, and strains. Major catastrophic injuries are rare. Overuse injuries secondary to repetitive, unrepaired microtrauma are becoming a more common source of injury. Children must not be thought of as simply small adults. Physiologically and emotionally, children, youth, and adolescents all vary in rate and magnitude of maturation in these spheres. The vast majority of sports-related injuries in children and adolescents require neither hospitalization nor surgery but lend themselves to out-patient treatment. The practice of preventive medicine is encouraged, and orthopedists should take an active part in advising patients about equipment and training programs to prevent coach and parental abuses that may lead to musculoskeletal long-term disabilities. PMID- 2971910 TI - Office management of low back pain. AB - Low back pain, with or without sciatica, is a very common medical problem. Since a great majority of patients can be successfully treated with nonoperative methods, office management of these patients is a very important issue for all clinicians. Patients with low back pain can be divided into three major groups: 1) the first group with acute (initial onset or very occasional recurrent) symptoms, 2) the second group with chronic (frequent intermittent recurrent or persistent) symptoms and 3) the third group with resistant symptoms. For the first group with acute symptoms, establishment of a specific diagnosis is rarely necessary. An active nonoperative treatment program (brief rest, protection, physical therapy modalities, medication, exercises, and reconditioning) provides a high rate (nearly 90 to 95 per cent) of success (return to normal activities and work) within 8 to 12 weeks. For the second group, with chronic symptoms, establishment of a specific diagnosis and a comprehensive and specific history are essential. Diagnostic procedures for specific pathologic conditions are outlined. A clear and specific treatment goal should be established and communicated to all the parties involved. Nonoperative treatment for this group includes back school, postural exercises, bracing, exercises for strength and endurance, work-site modification, vocational counseling, and pain management. For the third group with resistant symptoms, the role of the orthopedic or neurosurgeon is as a consultant to a pain management team for evaluation and treatment of missed diagnosis or complications of previous treatments. PMID- 2971911 TI - In vitro photosensitization of human head and neck squamous cancer cells by dihematoporphyrin-ether. AB - In vitro experiments were performed to evaluate the direct cytotoxic and photosensitizing effects of dihematoporphyrin-ether (DHE) on the head and neck squamous cancer cell line, UM-SCC-38. Normal fibroblasts, normal cultured keratinocytes, and the UM-MEL-1 pigmented malignant melanoma cell line were used as controls. The parameters of duration of light exposure, drug concentration, and incubation periods were studied. Uptake of DHE by squamous cancer cells, as assessed microscopically by intensity of fluorescence, was rapid and reached a dose-dependent maximum intensity within 10 minutes. Cells were irradiated with polychromatic light with an energy of one milliwatt/cm2 at a distance of 1.5 ft. Cells growing in plastic dishes were incubated in the dark for 1 to 4 hours with DHE in concentrations that ranged from 1.25 to 50 micrograms/ml. The cell monolayers were washed and irradiated for periods of time ranging from 15 to 120 min. Dose-dependent loss of cell viability could be detected by trypan blue dye uptake as early as 1 hour after radiation and continued to increase until 4 hours after light exposure. No further loss of cell viability was observed over the next 10 hours. There was no phototoxicity in the absence of DHE. UM-SCC-38 cells were more sensitive to the photosensitizing effects of DHE than were either normal fibroblasts or malignant melanoma cells. Recovery from the photosensitizing effects of DHE was observed if the DHE-containing medium was removed and the cells were incubated in the dark for periods that ranged from 1 to 14 hours before light exposure. UM-SCC-38 cells recovered more rapidly than normal fibroblasts, normal keratinocytes, or UM-MEL-1 cells. PMID- 2971912 TI - Comparison of lumbar paravertebral EMG patterns in chronic low back pain patients and non-patient controls. AB - According to myogenic models that relate abnormal EMG patterns to the experience of pain, lumbar paravertebral muscle activity has been considered to play an important role in chronic low back pain. In the present study, 40 chronic low back pain patients and 40 matched non-patient controls were compared on lumbar paravertebral EMG during mechanically stabilized static and dynamic postures. Differences between groups in lumbar curvature and spinal range of motion were determined using a dual goniometer technique. Although the two groups did not differ on absolute levels of EMG during quiet standing, significant differences were found for EMG patterns during dynamic postures. In addition, most patients did not show the flexion-relaxation response or the expected pattern of EMG responses during trunk rotation, most likely because of restricted range of motion and/or compensatory posturing. These findings provide support for the biomechanical model of chronic pain and indicate the need for further research pertaining to pain behavior and movement-related lumbar muscle activity. PMID- 2971913 TI - The premenstrual syndrome: psychophysiologic concomitants of perceived stress and low back pain. AB - The effects of experimentally induced and personally relevant stressors on low back EMG activity during 3 phases of the menstrual cycle in women with the premenstrual syndrome (PMS) and premenstrual low back pain were examined. Thirty nine women reporting either PMS and premenstrual severe low back pain (group 1), PMS with premenstrual moderate low back pain (group 2), or those reporting neither condition (group 3) participated. During each of 3 menstrual phase specific assessment sessions, participants were exposed to a neutral stimulus, an experimentally induced stressor, and 2 personally relevant stressors in a randomized order. Concomitant monitoring of low back EMG activity and heart interbeat interval was undertaken. Results indicated that participants in group 1 evidenced greater EMG changes in response to personal stressors compared to the neutral stimulus and experimentally induced stress during the premenstrual phase. EMG activity in response to personal stress was also significantly higher during the premenstrual phase than during the menstrual and intermenstrual phases for group 1, and higher than EMG changes evidenced by groups 2 and 3 during the same phase. Group 2, while not reporting as severe premenstrual back pain as did group 1, reported moderate levels of back pain and evidenced greater physiological reactivity to a personal stressor than did group 3. The findings highlight the link between personally relevant stressors and concomitant physiologic responsivity and the role that this arousal may play in the maintenance and exacerbation of premenstrual low back pain. PMID- 2971914 TI - Presentation of a new MMPI scale to predict outcome after first lumbar diskectomy. AB - In a pilot study MMPI was presented preoperatively to 57 patients operated for the first time for lumbar disk herniation. Six months and 24 months postoperatively, the patients were asked about their degree of pain, health status regarding back illness, and social functioning. An analysis of the individual MMPI questions in relation to outcome was performed, and on this basis we propose a new scale, the S.M. scale, which could predict poor outcome in either follow-up in 89% of the cases with poor outcome. PMID- 2971915 TI - Study of lymphocyte subpopulations in a group of patients with tuberculosis unresponsive to antibiotic treatment. PMID- 2971916 TI - The surfaces of the parasitic nematodes Trichinella spiralis and Toxocara canis differ in the binding of post-C3 components of human complement by the alternative pathway. AB - The binding of human complement components C3, C5 and C9 to the surface of the infective larvae of the nematode parasites Toxocara canis and Trichinella spiralis, by the alternative pathway, was examined by direct and indirect immunofluorescence on the intact parasites. This showed that although C3 bound to both nematodes, they differed markedly in the binding of C5 and C9; C5 bound only minimally to T. spiralis, and C9 binding to this parasite was barely detectable. In contrast, both early and late components bound to T. canis to a high density, comparable to, or in excess of, the binding of these components to the infective larvae of the trematode Schistosoma mansoni. The lack of binding of the post-C3 components to T. spiralis did not correlate with enhanced binding of the control protein, Factor H. PMID- 2971917 TI - Manipulative therapy. PMID- 2971918 TI - Pain clinic #10. Physical exercise in the treatment of low back pain. Part II: A practical regimen of stretching exercise. AB - In the first article of this three part series, research evidence for physical exercise in the treatment of low back pain were examined and data documenting significant therapeutic benefits was identified. However, different types and philosophies of exercise abound and, in clinical practice, little research evidence exists for substantiating one approach over another. This article will outline a practical physical stretching program with both empirical and research value in the treatment of over 4,100 patients with complaints of low back pain. PMID- 2971919 TI - Imaging rounds #96. Spondylolysis--laminal fragmentation on the AP view. PMID- 2971920 TI - [Diverse actions of pure crystalline progesterone, 17-alpha-hydroxyprogesterone capronate and medroxyprogesterone acetate (MPA), on the biosynthesis of collagen in the tail of the rat (ovariectomized females)]. PMID- 2971921 TI - Drug allergy. AB - Adverse reaction to drugs is a common problem in the pediatric population. This article discusses the different types of adverse drug reactions and their mechanisms. Also included are representative clinical syndromes of drug hypersensitivity reactions, as well as certain organ systems syndromes. Finally, diagnosis and management of allergic reactions to particular drugs such as penicillin, iodinated contrast dye, insulin, and others are discussed. PMID- 2971922 TI - Plasma atrial natriuretic peptide in patients with congenital heart diseases. AB - The plasma level of human alpha-atrial natriuretic peptide was measured in healthy children and patients, 1 month to 15 years of age, with congenital heart diseases. Significant increases were found in patients with a ventricular septal defect, tricuspid valve atresia, patent ductus arteriosus, and atrial septal defect but not in those with pulmonary valve stenosis or tetralogy of Fallot. The levels were significantly higher in children with ventricular septal defects (221 +/- 123 pg/mL) or patent ductus arteriosus (124 +/- 38 pg/mL) than in those with atrial septal defects (65 +/- 42 pg/mL) (P less than .01). The increased levels appeared to be correlated with enlargement of the left atrium (r = .85, P less than .01) but not with the right atrial size or the mean right atrial pressure. They were higher in younger than in older healthy infants, but this age difference did not affect the results. These findings indicate that human alpha atrial natriuretic peptide is released into the circulation in response to chronic atrial expansion in patients with congenital heart disease and may have an important role in volume homeostasis. PMID- 2971923 TI - [Prognostic aspects of immune disorders in glomerulonephritis in children]. PMID- 2971924 TI - [Phosphorus-calcium metabolism in urate nephropathies in childhood]. PMID- 2971925 TI - [Experience in the organization of medical care for newborn infants at a city maternity hospital]. PMID- 2971926 TI - [Organization of the treatment of stuttering in a logopedics kindergarten using the methods of therapeutic physical exercise]. PMID- 2971927 TI - A 15 year survey of Burmese amputees. AB - A 15-year retrospective study of 2228 civilian amputees was conducted at the Hospital for the Disabled, Thamaing, Rangoon. It was demonstrated that utilization of appropriate technology for development of essential components had enabled the hospital to serve more amputees. The ratio of male to female was 4.23: 1. The mean age was 31 years, male slightly older than female. Trauma was the leading cause of upper limb amputations (87%). In the lower limb although trauma (47%) was the prominent cause, disease (41%) was a close second. Major specific causes of trauma were gun-shot/explosion (25%), railway accident (20%) and road accident (19%). Leading specific causes of disease were leprosy (25%), vascular disease (24%) and gangrene (23%). Unless appropriate and effective preventive measures are instituted man-power drainage and demand for prosthetic services will continue. PMID- 2971928 TI - Rapid minipreparations of bacteriophage lambda DNA. PMID- 2971929 TI - Construction of a NotI linking library and isolation of new markers close to the Huntington's disease gene. AB - Linking clones contain sequences flanking recognition sites for enzymes cutting rarely in mammalian DNA. They can be used to obtain and correlate both physical and genetic mapping information over subregions of mammalian chromosomes. We have constructed and used a NotI linking clone library representing unmethylated NotI sites from HHW693 DNA, a hamster hybrid cell line containing 4p15-4pter and a fragment of 5p as its only human chromosome contribution. Human clones were identified by hybridisation with a cloned human repeat sequence, and localised further to subregions of human chromosome 4p15-4pter using a panel of additional hybrids. Clones from the region distal to the DNA probes (D4S10, D4S43, D4S95) linked to the Huntington's disease mutation, were further analysed. Four markers close to the HD gene: D4S111, D4S113, D4S114 and clone 417 are described here. In addition to serving as markers in physical and genetic mapping experiments, these linking clones provide probes next to cleavable NotI sites, and can therefore be used to screen NotI based chromosome jumping libraries. They also provide indications for potential gene sequences, identifiable as evolutionarily conserved sequences. PMID- 2971931 TI - The pediatrician, the disabled child, and the family: a special relationship. PMID- 2971930 TI - The effect of anaerobiosis on antistaphylococcal antibiotics. AB - The activity of eight antimicrobial agents which might be used in the treatment of staphylococcal osteomyelitis was tested under anaerobic conditions similar to those found in chronically infected bone. An agar-dilution method was employed to determine the minimum inhibitory concentrations of tobramycin, vancomycin, teicoplanin, ciprofloxacin, clindamycin, ceftriaxone, ticarcillin-clavulanic acid, and amoxicillin-clavulanic acid against 25 coagulase-positive and 25 coagulase-negative staphylococcal strains. The activity of tobramycin against coagulase-positive staphylococci, and of amoxicillin-clavulanic acid and ticarcillin-clavulanic acid against coagulase-negative staphylococci was markedly decreased with anaerobiosis. Vancomycin, teicoplanin, and ciprofloxacin were active against coagulase-positive and coagulase-negative staphylococci under both aerobic and anaerobic conditions. It was also found that antibiotic concentrations comparable to the high levels which might be achieved with local antibiotic therapy of osteomyelitis were not sufficient to overcome the level of resistance (100 micrograms/ml) of staphylococci which were not susceptible to tobramycin, clindamycin, ceftriaxone, and ticarcillin-clavulanic acid. PMID- 2971933 TI - Ethical issues in the care of newborn infants. PMID- 2971932 TI - Developmental assessment and early intervention programs for young children: lessons learned from longitudinal research. PMID- 2971934 TI - Harry. The price of being emotionally distant. PMID- 2971935 TI - Cardiac arrest associated with hypokalaemia in a patient receiving mithramycin. AB - The case is reported of a patient who developed severe hypokalaemia, leading to cardiac arrest, while receiving mithramycin for painful Paget's disease. The patient was also receiving antihypertensive and anti-anginal treatment including bendrofluazide, but plasma potassium before the administration of mithramycin was normal. Hypokalaemia has been reported in previous studies of the use of mithramycin but has received little attention. The drug should be used with great care in patients with cardiovascular disease and in those receiving diuretics. PMID- 2971936 TI - Extensive carbamazepine eruption with eosinophilia and pulmonary infiltrate. PMID- 2971937 TI - [Drug-induced pulmonary pathology]. PMID- 2971939 TI - [Is cholelithiasis a disease?]. PMID- 2971938 TI - Effects of monensin on the receptor-mediated endocytosis of 125I-labelled IgG by guinea-pig yolk sac in vitro. AB - The effects of the carboxylic ionophore, monensin, on the receptor-mediated binding and uptake of 125I-labelled IgG by the guinea-pig yolk sac have been studied in vitro. Exposure of tissue to 10 microM monensin resulted in a rapid inhibition of uptake which correlated with a time- and temperature-dependent loss of cell-surface receptor activity. Monensin appeared to bring about a change in receptor distribution since the lost activity could be detected after permeabilizing the tissue with saponin. Electron microscopic examination of monensin-treated tissue revealed that the apical plasma membrane of endoderm cells was depleted of coated and uncoated pits and that the apical cytoplasm contained numerous large vacuoles. Dilation of the Golgi apparatus was also observed. Normal surface receptor activity and ultrastructural features could be largely recovered by removal of monensin. Recovery of receptor activity was unaffected by the presence of cycloheximide. These results are consistent with a model in which IgG receptors are recycled and in which monensin blocks this process by causing receptors to be trapped intracellularly. Ammonium chloride or a combination of valinomycin and carbonyl cyanide p-(trifluoromethoxy) phenylhydrazone also brought about a loss of surface IgG receptors, lending support to the idea that inhibition of recycling was the result of perturbation of an intracellular acidification event and implying that passage through an acidic compartment may be important for correct receptor processing. PMID- 2971940 TI - [Statistics in daily practice. A surgeon's reflections]. PMID- 2971941 TI - [B virus, delta agent and human immunodeficiency virus infections in drug addicts]. AB - Infections with the B, D, A, NANB viruses and with the human immunodeficiency virus (HIV) are very common among drug addicts, some of whom may harbour several of these pathogens. The serum of 90 per cent of drug addicts contains one of the HBV markers, and 20 per cent of them carry an anti-D antibody which is more often present in HBs Ag-positive subjects but may also be found in those who are positive for anti-HBs and anti HBc antibodies. The presence of an anti-delta antibody increases the risk of severe histological lesions (chronic active hepatitis, cirrhosis), as does chronic alcoholism associated with drug addiction. Fifty to sixty per cent of drug addicts are seropositive for HIV. At the AIDS stage, hepatic lesions are extremely frequent (90 per cent), but they have low activity and are seldom responsible for death. PMID- 2971942 TI - [Heterozygote beta-thalassemia without microcytosis]. PMID- 2971943 TI - [Respiratory complications in status asthmaticus under mechanical ventilation]. PMID- 2971945 TI - [Full term pregnancy and Soulier-Boffa syndrome]. PMID- 2971944 TI - [Kaposi's sarcoma of the uterine cervix in a female patient with a heart transplant]. PMID- 2971946 TI - [Sex hormones and cortisol levels in patients with acquired immunodeficiency syndrome]. PMID- 2971947 TI - [Crossed digitalis-alkaloid detoxication with an anti-digoxin antibody. Surveillance and mixed elimination of circulating digitoxin]. PMID- 2971948 TI - [Legionnaires' disease in hairy cell leukemia]. PMID- 2971949 TI - [Considerable improvement of spasticity and dystonic phenomena by sulfamethoxazole-trimethoprim in Joseph's disease]. PMID- 2971950 TI - [Myasthenia associated with Horton's disease]. PMID- 2971951 TI - [Hemichorea, polycythemia and cyanogenic congenital cardiopathy. A case]. PMID- 2971952 TI - [Treatment of epilepsy. Superiority of a single daily dose of sodium valproate over fractionated doses]. PMID- 2971953 TI - [Increase in urinary excretion of vanilmandelic acid and noradrenaline under nifedipine]. PMID- 2971954 TI - [Secondary rectal lesions in abuse of dextropropoxyphene and paracetamol suppositories. 2 new cases]. PMID- 2971955 TI - [Use of deoxyribonucleic acid probes in infectious pathology]. PMID- 2971956 TI - [Tomographical use of Tc hexamethylpropylene amine-oxime. First experience (78 studies)]. AB - A new radiotracer of cerebral perfusion, 99mTc-labelled hexamethylpropylene amine oxime, has been tried in 78 subjects: 6 controls and 72 patients. Qualitatively, the distribution of this tracer in healthy subjects was very much the same as that obtained with a reference method using 133xenon inhalation. Quantitatively, there was no correlation between the real blood flow rate and the normalized cerebral uptake rate. On the other hand, the asymmetry indices obtained in controls (but also in 16 patients) correlated very closely with those obtained with 133xenon. Our first results in acute ischaemic diseases as well as in the evaluation of vasospasm or Alzheimer-like presenile dementia point to wide fields of application for the new compound. Unlike its predecessors, it is always available and will probably be used, without any logistic investment, with the standard equipment of all Nuclear medicine departments. PMID- 2971957 TI - [Injuries of the spleen. Value of an absorbable perisplenic mesh. 15 cases]. AB - Over a 2-year period, 15 patients with spleen injuries underwent a spleen repair using an absorbable perisplenic mesh. This procedure was the sole treatment in 11 cases, and it proved both simple and reliable. No major complication was observed. Eight patients could be followed up until the computerized tomographic images became normal. The absorbable perisplenic mesh is an important improvement, and in some cases it may replace other techniques for arresting splenic bleeding. PMID- 2971958 TI - [Diagnostic difficulties and current clinical aspects of Plasmodium falciparum malaria upon returning from a chemoresistance area]. AB - The incidence and clinical aspects of malaria in patients returning from chemoresistance areas were evaluated by means of two studies. The first one was a prospective study of 145 subjects back from a 4 months' stay in Gabon and who had complied with an appropriate chemoprophylaxis regimen. Twelve of these subjects were found to have particulate P. falciparum antigens on the 3rd day following their return. Spontaneous negativation of parasitaemia was observed in 10 cases, whereas 2 subjects were about to develop a malarial attack. Forty-five days after the return home, 5 new cases of malarial attack and 3 cases of anaemia had occurred, although none of these 8 patients had haematozoa on the 3rd day. In the second study 31 patients admitted to hospital in 1987 were reviewed. The following clinical syndromes were detected: malarial attacks with high parasitaemia in 6 cases, malarial attacks with low or undetectable parasitaemia in 5 cases, anaemia, thrombocytopenia or pancytopenia in 5 cases, and febrile alteration of the general condition in 3 cases. Two aspects of malaria were identified: "malaria infection" with a latent and asymptomatic parasitic infestation likely to disappear spontaneously or to turn into "malaria disease". The latter has protean manifestations, but bouts of fever with low parasitaemia and blood disorders are predominant. PMID- 2971959 TI - [Arterial hypertension, renal function and angiotensin-converting enzyme inhibition]. AB - Several effects of the antihypertensive treatment with angiotensin converting enzyme inhibitors (ACEI)--whether beneficial or detrimental--are best explained at the renal level. In the presence of a renal artery stenosis, the activation of the intrarenal renin-angiotensin system is first directed at maintaining glomerular filtration and intracapillary hydrostatic pressure through post glomerular vasoconstriction. Long-term elevation of blood pressure is associated with a shift of the renal function curve, possibly linked with an alteration of the intrarenal renin-angiotensin system. Inhibition of the converting enzyme does affect the mechanisms of sodium conservation. In a subset of essential hypertensive subjects, ACEI may correct a subtle primary abnormality in modulation by sodium of renal (and adrenal) response(s) to angiotensin. Finally, the possibility of renal protection in circumstances such as diabetes mellitus, provides an exciting area of investigations for antihypertensive treatment with ACEI. PMID- 2971960 TI - [Trendelenburg's operation using an sternostomy approach for pulmonary embolectomy]. AB - Five cases of extremely serious pulmonary embolism treated by embolectomy without extracorporeal circulation have prompted to recall the Trendelenburg's operation through sternotomy. Rapid, simple and requiring little equipment, this technique is perfectly suited to emergency surgery. Its use should save the lives of several patients who would be condemned to death without surgical operation. PMID- 2971961 TI - [Jaundice caused by tuberculous biliary stenosis. Percutaneous treatment]. PMID- 2971962 TI - [Bone and spinal tuberculosis: rare cause of false negative diagnosis with technetium scintigraphy]. PMID- 2971963 TI - [Vascular purpura caused by paracetamol. A case]. PMID- 2971964 TI - [Treatment of junctional tachycardia with high doses of flecainide in infants]. PMID- 2971965 TI - [Guided biopsy under endocavitary echography. Diagnosis of pelvic recurrence of anal canal cancer surgically treated]. PMID- 2971966 TI - [Prolonged toxic shock syndrome after cesarean section. A case]. PMID- 2971967 TI - [Is there a recrudescence of neurosyphilis? 7 cases observed in a 1-year period in non-carriers of the human immunodeficiency virus]. PMID- 2971968 TI - [Subacute intracranial subdural hygroma after lumbar puncture]. PMID- 2971969 TI - Plasma and urine concentrations of atrial natriuretic peptide in patients with diabetes mellitus. AB - To examine whether plasma and urine concentrations of human atrial natriuretic peptide (hANP) are altered in patients with diabetes mellitus (DM), plasma and urine hANP concentrations were evaluated in 86 patients with diabetes mellitus using an extraction procedure. The mean recovery rate of extraction was 71.8 +/- 0.6% (mean +/- SEM). The major immunoreactive component of hANP in extracted plasma and urine appeared to be identical to synthetic alpha hANP as judged by reverse-phase high-performance liquid chromatography (HPLC). The patients were divided into three groups according to their renal complications. The patients in group 1 had no apparent abnormality in serum creatinine, serum or urine beta 2 microglobulin (beta 2-MG), or urine N-acetyl-beta-D-glucosaminidase (NAG); those in group 2 showed either beta 2-MG or NAG abnormality but no creatinine abnormality. The patients in group 3 were though to have an established diabetic nephropathy and showed a serum creatinine increase. Plasma ANP concentrations in groups 1, 2, and 3 were 10.7 +/- 2.1, 19.9 +/- 5.6, and 39.2 +/- 9.9 fmol/ml, respectively. These values in groups 2 and 3 were significantly higher than the control values (p less than 0.05 or p less than 0.01 versus 6.2 +/- 0.7 fmol/ml). Urine ANP concentrations in group 1 were also within normal range, though those in groups 2 and 3 markedly increased in comparison with normal values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2971970 TI - Abdominal wall and gastric infarction in acute pancreatitis. AB - We report a case of severe acute pancreatitis associated with both gastric and abdominal wall infarction. The early skin changes on the anterior abdominal wall could be confused with Grey-Turner's sign. PMID- 2971971 TI - Identification of a functional mammalian spliceosome containing unspliced pre mRNA. AB - Functional 60S spliceosomes were assembled under conditions that block the first step of the mRNA splicing reaction. This block was imposed by carrying out the splicing reaction in nuclear extracts preincubated in 2.5 mM EDTA. Preparative amounts of the spliceosomes were isolated by gel filtration chromatography and shown to be functional by in vitro complementation assays. The unspliced pre-mRNA in the complex was converted to spliced products when incubated in cytoplasmic S100 extracts or in heat-treated or micrococcal nuclease-treated nuclear extracts. The latter result, in conjunction with the size of the complex, suggests that the spliceosome contains all of the small nuclear ribonucleoproteins (snRNPs) required for both steps of the splicing reaction. Biochemical characterization of the 5' cleavage reaction revealed that ATP and MgCl2 are required for this step in the splicing pathway. The presence of U1 snRNP in the blocked complex was demonstrated by quantitative immunoprecipitation of this complex by an anti-U1 snRNP monoclonal antibody. PMID- 2971972 TI - Adhesive receptor Mac-1 coordinates the activation of factor X on stimulated cells of monocytic and myeloid differentiation: an alternative initiation of the coagulation protease cascade. AB - Monocytes initiate coagulation through regulated surface expression of tissue factor and local assembly of a proteolytic enzymatic complex formed by tissue factor and factor VII/activated factor VII. We now show that, in the absence of these initiating molecules, monocytes and cell lines of monocytic/myeloid differentiation can alternatively initiate coagulation after exposure to ADP. The molecular basis for this procoagulant response consists of two distinct events. First, cell stimulation with ADP induces high-affinity binding of coagulation factor X to the surface-adhesive receptor Mac-1. Locally, Mac-1-concentrated factor X is then rapidly proteolytically cleaved to an active protease with size and activity characteristics of activated factor X, which supports the cell associated formation of thrombin and the procoagulant response. We conclude that the monocytic/myeloid adhesive receptor Mac-1 has the unexpected, specifically inducible property to organize a molecular assembly culminating in rapid fibrin formation that is independently regulated from tissue factor and factor VII/activated factor VII. PMID- 2971973 TI - A highly phosphorylated subpopulation of insulin-like growth factor II/mannose 6 phosphate receptors is concentrated in a clathrin-enriched plasma membrane fraction. AB - Insulin-like growth factor II (IGF-II)/mannose 6-phosphate (Man-6-P) receptors immunoprecipitated from purified plasma membranes of 32P-labeled rat adipocytes are markedly heterogenous in their phosphorylation state. Approximately 80% of the plasma membrane receptors are solubilized in 1% (vol/vol) Triton X-100 and are phosphorylated on serine residues at a stoichiometry of approximately 0.1-0.2 mol of phosphate per mol of receptor. In contrast, 15-20% of the receptors are Triton X-100-insoluble and are phosphorylated on serine and threonine residues at approximately 4 or 5 mol of phosphate per mol of receptor. This Triton X-100 insoluble membrane subfraction contains only 5% of the total plasma membrane protein and yet contains all of the clathrin heavy chain associated with plasma membrane, as detected by immunoblotting with a monoclonal antibody. Based on the relative yields of protein in the detergent-insoluble material, IGF-II/Man-6-P receptors are concentrated approximately equal to 3-fold in this clathrin enriched subfraction. Insulin treatment of intact cells increased the total IGF II/Man-6-P receptors in the Triton X-100-soluble fraction of the plasma membrane, whereas no change in receptor number in the detergent-insoluble fraction was seen. However, insulin markedly decreased the phosphorylation stoichiometry of the Triton X-100-insoluble receptors. Taken together, these results indicate that insulin decreases the phosphorylation state of a highly phosphorylated subpopulation of IGF-II/Man-6-P receptors on the plasma membrane. In addition, insulin action may prevent the concentration of these receptors in a clathrin enriched membrane subfraction. PMID- 2971974 TI - Complement receptor type three (CD11b/CD18) of human polymorphonuclear leukocytes recognizes fibrinogen. AB - Human polymorphonuclear leukocytes (PMN) have previously been shown to bind to aggregates of fibrin and to fibrinogen-coated surfaces. During their interactions with fibrinogen-coated surfaces, PMN make such close contact with the surface that a portion of the secreted elastase activity is protected from macromolecular protease inhibitors in the surrounding medium. Here we show that the receptor on PMN that mediates this interaction is complement receptor type 3 (CR3; CD11b/CD18), a molecule previously identified as a receptor for the complement protein fragment C3bi. Monoclonal antibodies against CR3 that block the binding of C3bi also block the binding of PMN to fibrinogen-coated surfaces and the formation of a protected compartment. The region of fibrinogen recognized by CR3 lies at the carboxyl terminus of the gamma chain, since peptides based on this sequence effectively inhibit the binding of PMN to fibrinogen-coated surfaces. These peptides also block the binding of C3bi-coated erythrocytes to CR3, thus indicating that a single binding site is used for binding both C3bi and fibrinogen. Sequence analysis shows strong structural similarity between this region of fibrinogen and other known ligands of CR3. These studies thus indicate that CR3 functions as a receptor not only for C3bi but also for fibrinogen. PMID- 2971975 TI - Side effects of flavonoids in medical practice. PMID- 2971976 TI - Plants causing adverse skin reactions. PMID- 2971977 TI - Comparison between aerosol and powder delivery system of fenoterol plus ipratropium bromide ('Duovent') in patients with asthma and chronic bronchitis. AB - A study was carried out in 15 stable asthmatics and 5 patients with partially reversible airflow obstruction to compare the efficacy and tolerability of a single dose of 200 micrograms fenoterol plus 80 micrograms ipratropium bromide ('Duovent') administered either by metered dose aerosol or as a dry powder preparation for inhalation ('Inhalets'). Using a double-dummy technique, patients received each formulation, in random order, on 2 separate days within 1 week. Spirometric measurements were made over a period of 6 hours after the dose. The results showed that a 35% peak improvement in mean FEV1 occurred with each method of administration and there was no statistical difference between responses. Improvements of 25% were sustained for over 4 hours and effects on pulse rate and tremor were similar for both preparations. PMID- 2971978 TI - The effect of 8-OH-DPAT on temperature in the rat and its modification by chronic antidepressant treatments. AB - Administration of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) to rats produced a dose-dependent hypothermia. Pretreatment with the receptor antagonist methiothepin abolished this effect, and pretreatment with haloperidol, propranolol and pindolol partially attenuated it, although methiothepin and pindolol had hyperthermic actions of their own. Other receptor antagonists including ritanserin, naloxone, clonidine, phenoxybenzamine and metergoline did not significantly modify the response elicited by subsequent 8-OH-DPAT challenge. In antidepressant studies, chronic treatment (22 days) with clorgyline attenuated the hypothermic response to 8-OH-DPAT, whereas similar duration of treatment with the tricyclics clomipramine and imipramine did not significantly modify it. Also, acute treatment for three days with each of the antidepressants did not modify 8 OH-DPAT-induced hypothermia. We conclude that rat rectal temperature can be a useful model to help assess the functional state of serotonergic mechanisms, including the adaptational changes induced by long-term antidepressant treatment. PMID- 2971979 TI - Intraspinalcord injection of colchicine as a tool to study micturition disturbances. Implication of endogenous opioid peptides. PMID- 2971980 TI - Physical therapists as first-contact nonphysician providers. PMID- 2971981 TI - The vascular anatomy of rectus abdominis musculocutaneous flaps based on the deep superior epigastric system. AB - Radiographic studies of the deep superior epigastric artery (DSEA) and its connections within the soft tissues of the abdominal wall were performed in 64 fresh cadavers. The patterns of anastomosis between the deep superior epigastric artery and the deep inferior epigastric artery (DIEA) were noted. Type I (29 percent) revealed a single deep superior epigastric artery and deep inferior epigastric artery, type II (57 percent) revealed a double-branched system of each vessel, and type III (14 percent) revealed a system of three or more major branches. In each case, the two systems were united by choke vessels in the segment of muscle above the umbilicus. The supply to the various transverse and vertical skin flaps from the deep superior epigastric artery was defined as a series of captured anatomic territories bounded by choke vessels. The upper transverse and vertical flaps had the best supply, and the TRAM flap had the most tenuous supply. Midline crossover occurs predominantly in the subdermal plexus and on the surface of the rectus sheath. Modifications of the design of the TRAM flap, the case for a delay procedure, the wisdom of including a strip of anterior rectus sheath, and the risks of splitting the muscle with respect to its nerve supply and vascular patterns are discussed on an anatomic basis. PMID- 2971982 TI - Mini-abdominoplasty. PMID- 2971983 TI - [Early and late results of percutaneous transluminal angioplasty of renal artery stenoses]. AB - The initial and long-term results of percutaneous transluminal angioplasty of renal artery stenoses are reported. In 4 out of 73 (5.5%) patients the angioplasty catheter could not be placed in the stenotic area. In 67 patients partial or complete dilatation of the segments treated was possible. Complications requiring therapy occurred in 12.3%. Most (80.3%) of the patients were discharged from hospital with completely corrected or improved hypertension. Cure or improvement was noted more often in patients with fibromuscular dysplasia, short history of hypertension, pathologic renin quotient, normal total hippuric acid clearance, or age under 40 years. On follow-up examination a mean of over 14 months after angioplasty 51.7% of the patients had normal blood pressure or less pronounced hypertension, while in 60.9% the total hippuric acid clearance had increased. PMID- 2971984 TI - Acute occlusion developing during or immediately after percutaneous transluminal coronary angioplasty: nonsurgical treatment. AB - The authors report results of nonsurgical treatment of acute occlusion developing during percutaneous transluminal coronary angioplasty (PTCA). Of 514 consecutive PTCAs, acute occlusion of the dilated artery developed during or within 1 hour after PTCA in 44 patients (8.5%); of these acute occlusions, 11 (2.1%) were due to guide wire or catheter manipulation during PTCA, while 33 (6.4%) were due to abrupt closure occurring after initially successful dilation. In seven of the 11 patients with acute occlusions resulting from manipulation, PTCA was continued with attempts to cross the reoccluded segment; this was successful in two patients (29%). In 25 of the 33 patients with abrupt closure, repeat balloon dilation was immediately attempted rather than send the patients to surgery for emergency coronary artery bypass grafting. Sixteen of these 25 attempts were successful (64%) and resulted in conversion of a potential complication into a successful outcome. Abrupt closure following initial dilation seems more likely to be successfully treated by continuing with PTCA than is acute occlusion resulting from catheter and guide wire manipulation. PMID- 2971986 TI - Plasma protein S deficiency and thromboembolic disease. PMID- 2971987 TI - Significance of pre-S2 peptide of hepatitis B virus: should it be in the vaccine? PMID- 2971985 TI - Estimation of human myocardial mass with MR imaging. AB - The accuracy and reproducibility of magnetic resonance (MR) imaging in the determination of left ventricular mass in humans was investigated. Left ventricular wall volume was measured from ten short-axis, end-diastolic MR images that spanned the left ventricle. Mass was estimated on the basis of average left ventricular wall volume and an assumed myocardial density. To establish the accuracy of the technique, the authors imaged ten cadaver hearts and compared true left ventricular weight with the mass estimate based on MR imaging findings. In vivo determination of left ventricular mass was evaluated in 40 subjects, with resultant calculated masses of 156.4-319.3 g. Intra- and interobserver variabilities of the technique were analyzed in ten subjects. Both the intra- (r = .96, standard error of estimate [SEE] = 11.1 g) and interobserver variabilities (r = .91, SEE = 17.8 g) were excellent. Eight subjects were imaged on two separate occasions to evaluate reproducibility of the technique and confidence limits for a given measurement. For these eight, there was good correlation between the two estimates (r = .93, SEE = 21 g). The authors conclude that MR imaging yields highly accurate and reproducible estimates of left ventricular mass in humans in vivo. PMID- 2971988 TI - Radiological patterns of lumbar spinal degenerative changes in symptomatic patients. PMID- 2971989 TI - [Percutaneous transluminal angioplasty of supra-aortic vessels. Presentation of 16 cases]. AB - Sixteen cases are reported of dilatation of supra-aortic vessels; in 14/16 patients the vessel involved was either the subclavian artery or the brachiocephalich trunk. Special attention is paid to the choice of patients--the ideal one presenting with a single uncalcified lesion, with stenosis more than 50% of diameter; the symptoms have recently appeared, with a significant difference (more than 20 mmHg) in the pressure of the two arms. The technical aspects of the angioplastic procedure are discussed, especially in order to preserve the intracranial circulation and to limit possible complications. The presence of reversed blood flow in the vertebral artery is extremely important to preserve intracranial circulation from possible embolism; normal flow obtained at the end of the procedure is therefore an useful proof of successful dilatation. The advantages are stressed of dilatation over the surgical techniques used in the past. Finally, the importance is emphasized of a correct follow-up and doppler control of supra-aortic circulation. PMID- 2971990 TI - [Percutaneous transluminal angioplasty of the vascular area of the lower limbs. Current indications]. AB - Usual indications for percutaneous transluminal angioplasty (PTA) in the vascular district of the lower limbs are single stenoses, short multiple stenoses and obstructions--shorter than 8 cm--in Fontaine's stage II, III and IV patients. Our experience is based on 642 PTAs carried out for obliterative arteriopathy of the lower limbs over a 6-year period. In agreement with the literature, we believe that today these indications can be extended to "limb salvage", to chronic femoral-popliteal obstructions longer than 8 cm and of a duration of less than one year, treated also with loco-regional fibrinolysis, and to asymptomatic lesions. Our series of patients include 15 treatments for "limb salvage", 21 chronic femoral-popliteal obstructions and 5 cases with bilateral lesions. The latter were treated with PTA only on the symptomatic side: later angiographic controls demonstrated a progressive deterioration only on the asymptomatic side. Immediate results were positive in 86% of the "limb salvage" cases and in 85% of the chronic femoral-popliteal obstructions. The advantages are discussed of PTA in the above-mentioned cases, paying also attention to the cost-benefit ratio. PMID- 2971991 TI - [Pelvic inflammations]. PMID- 2971992 TI - [Clinical observations on the cost-effectiveness relationship in endoscopic surgery on the lower urinary tract]. PMID- 2971993 TI - [Prognostic factors in reinterventions in biliary lithiasis]. PMID- 2971994 TI - [Upper digestive hemorrhages in the elderly: pathology, evolution, treatment]. PMID- 2971995 TI - [Papillary operations in biliary reinterventions]. PMID- 2971996 TI - [Use of free de-epidermized skin as autograft material in the surgical repair of umbilical hernias, eventrations and other diseases]. PMID- 2971997 TI - [A special method of extrahepatic biliodigestive anastomosis]. PMID- 2971998 TI - [Wounds of the portal vein due to silent weapons]. PMID- 2971999 TI - [Hyperchloremic metabolic acidosis during postoperative total parenteral nutrition]. PMID- 2972000 TI - [Acute postoperative necrotizing enteritis]. PMID- 2972001 TI - [Principles of resuscitation and anesthesia in early postoperative obstructions]. PMID- 2972002 TI - [Complications of venous catheterization]. PMID- 2972003 TI - [Emergencies in chemical eye burns]. PMID- 2972004 TI - [Ophthalmological changes in rheumatoid arthritis]. PMID- 2972005 TI - [Glaucoma secondary to aphakia and its outpatient therapeutic management]. PMID- 2972006 TI - [The role of ultrasonography in the implantation of an artificial crystalline lens]. PMID- 2972007 TI - [Implantation of an artificial crystalline lens in eyes with Fuchs' syndrome]. PMID- 2972008 TI - [Implantation of an inverse artificial crystalline lens]. PMID- 2972009 TI - [Reimplantation of an artificial crystalline lens]. PMID- 2972010 TI - [Idiopathic microcystic macular edema]. PMID- 2972011 TI - [A case of anophthalmos]. PMID- 2972012 TI - [Massive-focus choroiditis in staphylococcal septicemia with multiple sites]. PMID- 2972013 TI - [Persistent oculocardiac reflex following retrobulbar hematoma. Case report]. PMID- 2972014 TI - [History of the correction of vision from antiquity to our time]. PMID- 2972015 TI - [Cyclosporin A in ophthalmology]. PMID- 2972016 TI - [Viral keratitis: diagnostic and treatment principles]. PMID- 2972017 TI - [Phacolytic glaucoma]. PMID- 2972018 TI - [Percutaneous transluminal angioplasty in hypertensive patients with stenosis of the renal arteries]. PMID- 2972019 TI - [Hepatorenal syndrome]. PMID- 2972020 TI - [Coronary angioplasty in total occlusions]. PMID- 2972021 TI - [Multiple coronary angioplasty]. PMID- 2972022 TI - [Hepatitis A in the adult during vaccination against hepatitis B]. PMID- 2972023 TI - [Abdominal non-Hodgkin's lymphoma diagnosed by laparoscopy]. PMID- 2972024 TI - [Adverse clinico-humoral manifestations in ulcer patients treated with cimetidine. Our experience in 488 patients]. PMID- 2972025 TI - Plasma and CSF levels of immunoreactive beta-endorphin in algic peaks of patients with herniated intervertebral discs. AB - Plasma and CSF levels of beta-Endorphin (beta-End) were measured by radioimmunoassay in three groups of human subjects. The first group consisted of healthy adults, and only plasma beta-End was determined. The second group consisted of patients showing non-painful neurological diseases. The third group consisted of patients suffering from acute pain due to herniated intervertebral discs. In the last two groups, beta-End levels were measured in plasma and CSF. The results showed that plasma levels of beta-End were similar in the first two groups of patients. In contrast, patients with acute pain showed significantly increased levels of beta-End in plasma. CSF levels of beta-End did not show significant differences among the groups. The results suggest that the increase in plasma levels of beta-End was a consequence of the stress produced by acute pain. PMID- 2972026 TI - Serum androgen levels after streptozotocin administration in the male rat. AB - Serum levels of testosterone and dihydrotestosterone were measured in control and diabetic animals 5, 10 and 15 days after streptozotocin administration. The diabetic state produced a marked reduction in serum androgen levels 10 and 15 days after streptozotocin administration. Insulin treatment partially restored the circulating androgen levels when administered to diabetic rats. PMID- 2972027 TI - [Platelet activation in vivo before and after hemodialysis using polyacrylonitrile filters]. PMID- 2972028 TI - [Laparoscopy in the staging of lymphoma. Personal experience]. PMID- 2972030 TI - [At the midpoint of the United Nations Decade of the Handicapped 1983-1992]. PMID- 2972029 TI - Interactions of hyperthermia with hypoxic cell sensitisers. PMID- 2972031 TI - [Synopsis of procedures used in the evaluation of the physically handicapped and work demands]. AB - The selection of workplaces for disabled people requires the matching of their abilities with the requirements of potential workplaces. Such a comparison may occur in particular by directly contrasting abilities and job demands, for instance by profile comparison. Abilities are thereby described in terms of job demands and recorded in standardized items. In a synopsis of 13 existing job/person evaluation techniques it was to be ascertained which items are used for assessment of abilities and job demands and how detailed the various sections of abilities and job demands are recorded. For reasons of perspicuity the items used in the various techniques were grouped in 10 different sections. It turned out that the sections "work environment" and "movements of parts of the body" were in general emphasized most. Finally, the different sections are emphasized and subdivided differently in the various techniques, according to their scope (such as employment agencies, personnel planning, job design) and their range of applicability (to one company/different companies, to different branches of industry, etc.). The criteria of universality, completeness and acceptance are discussed in terms of devising an assessment technique. PMID- 2972032 TI - [Community-oriented special education requirements for handicapped persons in India]. PMID- 2972033 TI - Atrial natriuretic peptide, arginine vasopressin, and the renin-angiotensin system in carbon tetrachloride-induced hepatitis in rats. AB - The concentrations of atrial natriuretic peptide (ANP), arginine vasopressin (AVP) and hormones of the renin-angiotensin axis were studied in male rats with carbon tetrachloride-induced hepatitis and the results compared to normal control animals. The rats with hepatitis exhibited lower concentrations of ANP, plasma renin activity (PRA), and angiotensin I (AI) than did the control animals. Plasma concentrations of AVP and aldosterone were not significantly different in the two groups. The results suggest that experimental hepatitis is associated with renal hyperperfusion together with reduction in atrial pressures. PMID- 2972034 TI - Changes in abdominal muscle length during breathing in supine dogs. AB - To assess the mechanical function of the abdominal muscles during eupnea in the dog, we have measured the electrical activity and the respiratory changes in length of the rectus abdominis, external oblique, and transversus abdominis muscles in eight supine, lightly anesthetized, spontaneously breathing animals. Seven animals had phasic expiratory electromyographic (EMG) activity in the transversus and showed expiratory shortening of the muscle below its in situ relaxation length (Lr). In contrast, only three animals had expiratory EMG activity and expiratory shortening of the external oblique, and no animal had expiratory EMG activity in the rectus. Seven animals, however, showed shortening of the rectus muscle during expiration. The amount of transversus expiratory shortening in the eight animals averaged (mean +/- SE) - 7.61 +/- 1.72% Lr and was significantly larger (P less than 0.005) than the amount of external oblique (-0.11 +/- 0.10% Lr) or rectus (-0.90 +/- 0.39% Lr) expiratory shortening. Hyperoxic hypercapnia amplified these differences. These data thus indicate that in supine anesthetized dogs (1) the transversus is, in real mechanical terms, the primary abdominal muscle of expiration; and (2) the abdominal compartment of the chest wall during eupnea moves both below and above its neutral position, and not exclusively above it. PMID- 2972035 TI - Low back syndromes. PMID- 2972037 TI - Neurophysiologic evaluation of patients with low back pain. AB - 1. Neurophysiologic testing is as accurate as CT scanning or myelography in the diagnosis of root compression due to disk disease. 2. The most reliable findings are abnormal H reflex, and positive sharp waves or fibrillations, or both, in the appropriate muscles. 3. Neurophysiologic testing may be more accurate than clinical examination or radiologic study in patients with canal stenosis. 4. Symptomatic post-laminectomy patients present a difficult diagnostic problem, even when all available methods are used. PMID- 2972036 TI - Computed tomography in the evaluation of acute and chronic low back pain syndromes commonly seen. AB - The purpose of this article has been to provide the results of a survey of the more common conditions, and their CT findings, causing chronic low back pain. There is an extensive literature on this subject, providing much more detail and discussing less common entities, of which the reader may avail himself. PMID- 2972038 TI - Clinical evaluation of patients with low back pain. PMID- 2972039 TI - Treatment and functional restoration of the patient with back pain. PMID- 2972040 TI - A medical orthopaedist talks about low back syndromes. PMID- 2972041 TI - An orthopaedist talks about low back syndromes. PMID- 2972042 TI - A rehabilitation counselor talks about low back pain. PMID- 2972043 TI - Management of chronic low back pain: a comprehensive approach. AB - The treatment approach presented in this article is an obvious departure from ways physicians are typically trained to handle patients' pain complaints. Traditional medical training focuses primarily on the management of acute pain. Unfortunately, the treatment modalities appropriate for acute pain are not applicable to most chronic pain disorders. Since physicians' practices contain many chronic pain patients, it is important for them to develop a more comprehensive and effective approach to the management of CLBP. Through the use of case vignettes, this article has attempted to elucidate some common problems experienced by patients with CLBP. It is a complex disorder that requires that the physicians be sensitive to the biologic, psychologic, and social aspects of the illness. Simply handing a patient with CLBP a set of back exercises or prescriptions for narcotics and sedatives will not be beneficial. Rather, the patient must be educated about the pain and taught to take an active role in his own treatment. By working with patients and their families, physicians can teach patients with CLBP the self-management skills essential for the resumption of a normal, productive life. PMID- 2972044 TI - [Genetic-familial data in the major affective diseases]. PMID- 2972045 TI - [Computed tomography in a study to localize aphasic symptoms]. PMID- 2972046 TI - [Morphological characteristics and the evolutive electroclinical aspects of pathological biopotentials in subacute sclerosing panencephalitis]. PMID- 2972047 TI - [Electroretinographic studies in different lesions of the brain]. PMID- 2972048 TI - [The therapeutic approach in the carpal tunnel syndrome]. PMID- 2972049 TI - [The occupational recuperation of schizophrenics]. PMID- 2972050 TI - [Imipramine effects at the genetic level studied in vitro in man and in vivo in mice]. PMID- 2972051 TI - [Dr. Constantin Daniel (1914-1987)]. PMID- 2972052 TI - [Prions]. PMID- 2972053 TI - [Possible taxonomic description of coagulase-negative staphylococci isolated in infectious skin processes]. PMID- 2972054 TI - Evaluation of the systemic anticholinergic activity of nasally administered ipratropium bromide. AB - Plasma concentrations of nasally inhaled ipratropium bromide were analyzed in eight healthy volunteers by using a sensitive radioreceptor assay (RRA). The rate of saliva secretion, heart rate and changes in visual accommodation were quantitated in order to measure possible systemic anticholinergic drug effects. 240 micrograms of ipratropium bromide (40 micrograms each nostril, repeated twice at 15 min intervals) were inhaled nasally in a double blind, randomized, placebo controlled experiment. Ipratropium bromide absorbed fast, and peak plasma concentrations of the drug (257 +/- 55 pg/ml) were detected as soon as 5 min after the last inhalation (at 35 min from the beginning). The plasma levels of ipratropium bromide decreased rapidly, being only 86 +/- 7 pg/ml one hour after the last inhalation. These low concentrations of ipratropium bromide indicate that only a small portion of it absorbs after nasal application, which is consistent with the lack of any systemic anticholinergic drug effects in our subjects. It is concluded that nasally applied ipratropium bromide is not likely to cause systemic anticholinergic side-effects, even in doses exceeding the therapeutic recommendations. PMID- 2972055 TI - [Disk collapse after nucleolysis with chymopapain]. PMID- 2972056 TI - [Irreducible lordosis due to lamina-dural fusion]. PMID- 2972057 TI - [Cellular targets of immunoglobulins eluted from the placenta]. AB - Placenta eluted gammaglobulin were reported to have a beneficial effect among rheumatoid arthritis suffering patients. These gammaglobulin obtained by acidic elution from washed placentas (PEGG) were compared to plasmatic immunoglobulin (Ig) for their ability to inhibit mixed lymphocyte reaction (MLR), for their content of antibodies (Ab) directed against human tissues or bacteria and viruses. In addition, biochemical fractionations of PEGG were performed. Both PEGG and Ig from plasma (Sandoglobulin [SG]) contained anti nuclear Ab, Ab against smooth muscle, epithelium, vascular endothelium, synovial cells from rheumatoid arthritis synovium. Furthermore, PEGG at high concentration stained the Raji cell line that SG did not stain. PEGG inhibited undirectional MLR by acting on the stimulating cells. SG or plasmin digested Ig from retroplacental blood (Veinoglobulins, Merieux [VG]) also inhibited MLR but at 20 to 50 fold higher concentrations than PEGG. This activity was restricted to intact IgG, even further purified on protein A. Biochemical fractionation indicated that about 10% of PEGG consisted of free light chains, suggesting that deposits of free light chains might exist on placentas. PMID- 2972058 TI - [Abdominal-crural contracture in flexion related to hypophyseal insufficiency]. PMID- 2972059 TI - Pharmacological criteria for risk-benefit evaluation of NSAIDs. AB - Evaluation of new non-steroidal anti-inflammatory drugs (NSAIDs) must compare efficacy and toxicity with existing compounds. Real progress involves maintaining effectiveness while decreasing toxicity. It is relatively easy to assess the effects of NSAIDs in animal models, and to determine gastrointestinal toxicity. However, although the ratio of active and toxic doses in animals can be extrapolated to man, the approach is limited and the NSAID needs to be assessed in a clinical setting as early as possible. In France, a national survey system has reported a wide range of adverse effects related to NSAIDs and shown important differences between compounds. Overdosage may be one of the factors responsible for toxicity, therefore pharmacokinetic evaluation is useful. In some disease states e.g. rheumatoid arthritis, there is a higher possibility of saturation pharmacokinetics with some drugs. Other pharmacokinetic parameters of interest are half-life, functions limiting activity, and hepatotoxicity. Furthermore, different pharmacokinetic parameters are required for different forms of disease. In acute states, the NSAID should have a short half-life and low protein binding and vice versa in chronic states. An important goal is to develop more selective NSAIDs regarding mechanisms of action or distribution into diseased tissues. PMID- 2972061 TI - Alzheimer's proteins. PMID- 2972060 TI - [Spinal claudication following ileocaval venous thrombosis]. AB - The case is reported of a young male who presented with intermittent claudication after thrombosis of the right common femoral, iliac and caval vein. The symptoms were explained by the severe proximal venous obstruction. The patient, however, also complained of lumbar pain during and long after walking, and showed signs of irritation of nerve roots L4 to S1 on the right side. Venography and CT-scan disclosed venous collateral circulation leading through the spinal canal. The blood drained through the foramina intervertebralia to the caval vein, causing compression of the nerve roots depending on their filling. Thus, the claudication was "venous" in two ways, muscular (due to inadequate drainage) and spinal (due to intermittent compression of lumbar nerve roots by the collateral circulation). PMID- 2972062 TI - Virtue in viruses. PMID- 2972063 TI - "Snurps". AB - The whimsical acronym refers to "small nuclear ribonucleoproteins," which play a key role in the process whereby genetic information directs the activities of a living cell. SnRNP's help to delete the meaningless stretches called introns from messenger RNA. The author was one of the first investigators to reveal their function. PMID- 2972064 TI - The synthesis of protective fragments of alpha-human atrial natriuretic polypeptide (alpha-hANP). AB - In this paper the synthesis of alpha-human atrial natriuretic polypeptide fragments (positions, 1-4, 5-10, 11-16, 17-22, 23-28) by solution method is reported. In the synthesis of some peptides, the results from mixed anhydride procedure and active ester procedure are compared. In the synthesis of other peptides, the effects of reaction temperature, basicity of catalyst, and reaction time on the specific rotations and yields of these compounds are observed. The sequence of amino acids of all the protective polypeptides is confirmed by FAB MS. PMID- 2972065 TI - Antibody-enhanced infection by HIV-1 via Fc receptor-mediated entry. AB - Monocytes and macrophages, which may play a central role in the pathogenesis of infection with human immunodeficiency virus type 1 (HIV-1), express the CD4 molecule and Fc receptors (FcR) for immunoglobulin G (IgG). To explore the possibility that FcR mediate HIV-1 infection of monocytes, studies were conducted with the human monocytic cell line U937. These cells were exposed to HIV-1 complexed with various concentrations of serum from HIV-1 antibody-positive individuals and monitored for HIV-1 replication. Serum samples from antibody negative normal individuals did not affect virus yields. High concentrations of antibody-positive sera showed virus-neutralizing activity; however, cells infected with HIV-1 in the presence of antibody-positive sera at subneutralizing concentrations significantly enhanced virus replication. This infection enhancement was blocked by heat-aggregated gamma-globulin. Moreover, the IgG fraction from an HIV-1 antibody-positive serum enhanced HIV-1 infection at the same serum dilution equivalents. In contrast, IgG-F(ab')2 did not enhance HIV-1 infection but showed neutralizing activity with HIV-1. These results are compatible with the concept of FcR-mediated infection enhancement and suggest that this immunological response to HIV-1, instead of protecting the host, potentially facilitates the infection. PMID- 2972066 TI - Lipoprotein(a): a genetically determined lipoprotein containing a glycoprotein of the plasminogen family. AB - Lp(a) represents a genetically transmitted class of plasma LDL having apo B-100 linked by a disulfide bridge to a glycoprotein, apo(a). Lp(a) is heterogeneous in size and density. Apo(a) is also heterogeneous in size (molecular weight between approximately 300,000 and 700,000) due probably to the polymorphism of both polypeptide and carbohydrate chains. Recent studies have shown that apo(a) has a striking amino acid sequence homology with plasminogen, a serine protease zymogen that following activation to plasmin enters the fibrinolytic system. Apo(a) is severalfold larger than plasminogen (molecular weight approximately 90,000) and also differs from it because it fails to be activated to plasmin. This is due to the fact that arginine is replaced by serine at the site of cleavage by streptokinase, urokinase, or tissue plasminogen activator. A single gene locus appears to control the Lp(a) polymorphism as well as the concentration of the Lp(a) phenotypes in the plasma. Patients with high plasma levels of Lp(a) have been shown to have an increased incidence of cardiovascular disease but a causal relationship has not been firmly established. The information that is being rapidly acquired on the structure of Lp(a) should facilitate the understanding of the molecular basis of the polymorphism of this genetic variant and of the role that the various Lp(a) phenotypes play in atherosclerosis and thrombosis. The potential physiologic role of Lp(a) remains open to inquiry. PMID- 2972067 TI - Effects of n-3 fatty acids on reocclusion after angioplasty. PMID- 2972068 TI - Radionuclide evaluation of renal transplants. AB - In this review article, the following topics are treated: the radiopharmaceuticals 99mTc-diethylenetriaminepentaacetic acid (DTPA), 131I orthoiodohippurate (OIH), 99mTc-mercaptoacetyltriglycine (MAG3), 67Ga-citrate, radioiodinated fibrinogen, 99mTc-sulfur colloid, 111In-labelled white cells and platelets; gamma camera methods based on images, on first pass and on tubular transit; blood clearance methods; and the diagnosis of surgical complications, acute rejection (AR), acute tubular necrosis (ATN), chronic rejection (CR), and cyclosporine-A (CYA) toxicity. PMID- 2972069 TI - Journey patterns of the elderly and disabled in the Cotswolds: a spatial analysis. AB - The decline of services and transport provision in the North Cotswolds has led to a progressive deterioration in the welfare of the elderly and disabled. This study, based on interviews with 498 individuals in eight typical parishes, relates the travel behaviour of these groups to health status, personal circumstances, the grade and location of services, and mode of transport. The repercussions of the 1985 Transport Act for the car-less and severely disabled are then examined. Given the increased loadings on domiciliary support systems, the case is argued for coordinated initiatives in alternative transport provision which relate, sensitively, to the needs of the different groups among the elderly and disabled. PMID- 2972070 TI - Correlations of hip mobility with degree of back pain and lumbar spinal mobility in chronic low-back pain patients. AB - Mobility of hips and lumbar spine were measured in 301 men and 175 women who were in employment but suffered from chronic or recurrent low-back pain. The degree of low-back pain (LBP) was assessed with a questionnaire. Hip flexion, extension, internal rotation, and hamstring flexibility in the men, and hip flexion and extension in the women had statistically significant negative correlations with LBP. Among the correlations between hip and lumbar spinal mobility, hip flexion and extension with lumbar rotation were strongest. PMID- 2972071 TI - Annular protrusion: pathophysiology and roentgenographic appearance. AB - The degenerative changes of the lumbar spine in 100 patients with symptomatic low back pain were reviewed using plain roentgenograms and computed tomographic scans in order to determine the relationship between degeneration and annular protrusion. Additionally, the lumbar spinal units of 25 fresh cadavers were roentgenographed, injected with a mixture of methylene blue and renografin-60, dissected, and studied. The state of degeneration of each of the intervertebral units of both groups was graded on a four-point scale based on the roentgenographic presence or absence of osteophytes and facet joint changes, and the intervertebral disc height. The degree of annular protrusion was measured by dividing the anteroposterior diameter of the intervertebral disc by the anteroposterior diameter of the vertebral plate as determined on the radiographic studies. In the clinical group, 39 intervertebral discs having Grade II and III degeneration exhibited a statistically higher annular/vertebral diameter ratio (A/V index) of 1.30 as compared to the normal invertebral discs, with an A/V index of 1.12 (P less than 0.001). The dissection of the disc spaces of the cadavers with radiographic evidence of disc space narrowing and marginal osteophytosis, Grade II and III degeneration, displayed significant interruption and tearing of the annular fibers and peripheral migration of collagenized nuclear fragments. In both the clinical and pathologic groups, there was a direct correlation between the size of the annular bulge, as indicated in the A/V index, and the degree of narrowing of the disc space. Therefore, this study indicated that annular protrusion is an intricate part of the degeneration process. PMID- 2972072 TI - Evaluation and outcome of low-back pain of unknown etiology. AB - Between January 1980 and January 1985, 5,362 patients with low-back pain were evaluated prospectively using a standard approach. The majority improved with routine therapy or could be placed in a definitive diagnostic group. One hundred and nine (2%), however, failed to improve or could not be assigned a specific diagnosis. These patients, defined as having chronic low-back pain of unknown etiology, were referred to a rheumatologist for evaluation. The average time to referral was 6 months. A specific diagnosis was obtained for 14 of the 109 patients. The remaining 95 were placed on new treatment plans, which included detoxification, a change of physicians, and arbitrary changes in therapy. The results indicated that approximately 75% of the group had 50% or greater reduction of pain, and half of the workers' compensation patients returned to some form of employment (although perhaps only intermittent). PMID- 2972073 TI - The use of pain drawings in screening for psychological involvement in complaints of low-back pain. AB - The pain drawings of 54 low-back-pain patients were examined to find out if it is possible to use them as a brief screening test in order to assess the psychological impairment of the patients. We were using the scoring system of Ransford et al, which we slightly changed, and chose as a criterion variable the ERMSS (Erweiterte Revidierte Mehrdimensionale Schmerzskala) of Cziske. This test originates in the McGill Pain Questionnaire of Melzack and Torgerson; its scales describe four dimensions of pain perception: pain intensity, the sensory discriminative dimension reflecting the somatic aspect of pain; the affective motivational dimension, and the total number of words, both representing the psychological involvement of a pain patient. A correlation was found between pain drawing score and the sensory-discriminative dimension of pain perception, whereas there was no such correlation between drawing score and the affective dimension. These results indicate that the pain drawing score might not be a sufficiently valid instrument for assessing psychological disturbances in pain patients to allow it to be used for individual diagnosis without hesitation. PMID- 2972074 TI - Computed tomography and fluoroscopy guided anesthesia and steroid injection in facet syndrome. AB - One hundred patients with posterior compartment lumbar spinal axis pain syndromes and focal tenderness were injected with lidocaine and betamethasone in 194 sites. Lidocaine injection was diagnostic in 183 instances (94%) and steroid injection provided long-term relief (greater than 3 months) in 105 instances (54%). CT guidance proved helpful in directing the needle tip to the precise location for optimal delivery of medications to exact anatomical sites. PMID- 2972076 TI - A technique for early wound management using polypropylene mesh reconstruction of the abdominal wall. PMID- 2972075 TI - Levels of keratan sulfate-bearing fragments rise predictably following chemonucleolysis of dog intervertebral discs with chymopapain. AB - Chymopapain (1 mg) was injected into each of four-lumbar intervertebral discs of adult mongrel dogs. As expected, at 2 weeks, all injected discs exhibited marked loss of height (mean: 50% of original height) indicative of severe proteoglycan depletion. The appearance of keratan sulfate-bearing fragments in plasma was monitored by an ELISA-inhibition assay which uses a monoclonal antibody (1/20/5-D 4) specific for an epitope present only in the longest keratan sulfate chains. Levels of plasma keratan sulfate rose within 30 minutes and reached a maximum between 24 and 72 hours later. Levels then declined progressively but were still elevated at 2 weeks postinjection. Keratan sulfate-bearing fragments in plasma were purified by ion exchange chromatography on DEAE-Sephacryl and fractionated by sieve chromatography on Sepharose CL-6B. These plasma keratan sulfate-bearing fragments were found to be similar in size to keratan sulfate-bearing fragments generated by chymopapain digestion of dog nucleus pulposus proteoglycans, but slightly larger than single keratan sulfate chains obtained by alkaline borohydride treatment of dog nucleus pulposus proteoglycans. The results of this study show that measurements of blood levels of keratan sulfate could prove useful in monitoring effective degradation of disc proteoglycans in chemonucleolysis in man and help discriminate between ineffective enzyme placement, and alternative mechanisms of treatment failure. PMID- 2972077 TI - Diagnostic and therapeutic laparoscopy. An entity often overlooked by the surgeon. AB - For unknown reasons, only a few surgeons have become interested in diagnostic and therapeutic laparoscopy. Electronic imaging, the recent adjunct to endoscopy, has enhanced its value. In problematic cases, it is not only important to establish the diagnosis but also to be able to demonstrate the interesting findings during weekly conferences or to reevaluate or recapitulate the findings in the follow-up period. Ascites, pyrexia, abdominal pain with weight loss of unknown origin, a palpable mass, suspected primary or secondary liver tumors, staging, second look, and questionable operability of known primaries are examples of the indications for laparoscopy. If intra-abdominal malignancy is suspected, a computed tomographic (CT) scan or ultrasonic examination is the common current procedure. If liver or parietal peritoneal involvement is present, these costly examinations will not display lesions smaller than 1 cm, whereas they can often be seen during laparoscopy. Repeated CT or ultrasound examinations with guided needle biopsies have a high incidence of noninformative (cytological) specimen retrieval. Laparoscopy, providing a precise, well-aimed biopsy, can be performed under local anesthesia following premedication. It represents an important advantage for high risk patients because it can be performed with a negligible incidence of complications. PMID- 2972078 TI - Clinical evaluation of laser endoscopy for the treatment of gastric tumors. AB - Yttrium aluminum garnet (YAG) laser therapy was performed through a gastroscope on 123 patients from 1980 to 1986. A complete cure with YAG laser therapy was obtained in patients with atypical epithelium of gastric mucosa and gastric polyps. Laser therapy (YAG) was very effective for the elevated type of early gastric cancer, and radical treatment can be expected for well-differentiated adenocarcinoma of less than 2 cm when tumor infiltration does not exceed the mucosa. YAG laser therapy for the depressed type of gastric cancer was not as effective and presents various problems. Argon-dye laser therapy, however, is more effective for the depressed type of gastric cancer. Endoscopic laser therapy is very effective, but it is necessary to be aware that lymph node metastases of gastric cancer have been found frequently, even in the early stage. Therefore, laser therapy must be performed under strict indications. PMID- 2972079 TI - The clinical significance of coronary angioscopy in patients with coronary heart disease. AB - The feasibility and safety of coronary endoscopy was evaluated in three sets of investigations: in 7 cadaver hearts, in 11 patients undergoing coronary bypass surgery, and in 30 patients during routine cardiac catheterization prior to coronary balloon angioplasty (PTCA). In three of the seven cadaver hearts the lumen of the arteries appeared normal. In three diffuse atherosclerotic lesions, and in one, a high-grade, tight stenosis were observed. In nine of eleven patients in the operation room, the lesions of interest could be visualized, and high-grade stenoses were found in all. In addition, in three patients with unstable angina pectoris, fresh thrombi were seen at the site of stenosis. In six of the nine patients, the periphery of the native coronary vessel was found to have no further stenotic regions. During cardiac catheterization in 17/30 patients, the lesion of interest could be examined angioscopically, and in 13 instances the stenosis appeared excentric and irregularly shaped. In three instances, multiple ulcerations were seen in the stenotic area. In two of the five patients, intimal ruptures were found following PTCA, which could not be documented angiographically. Coronary endoscopy provides valuable additional information on the nature and appearance of atherosclerotic lesions. It can be performed clinically without great harm to the patients. Despite some limitations, it will probably become a routine diagnostic tool in patients undergoing routine coronary angiography, balloon angioplasty or high-frequency angioplasty, and coronary bypass grafting. PMID- 2972081 TI - [Work environment. Occupational injury and economics]. PMID- 2972080 TI - Peritoneoscopy for the diagnosis of acute appendicitis in females of reproductive age. AB - The diagnosis of acute appendicitis in women of reproductive age can be a difficult clinical problem. A retrospective audit found our incidence of normal appendix removal in this patient population to be 42%. This study was undertaken to determine if the selective use of peritoneoscopy would improve the accuracy of diagnosis and thereby reduce the need for celiotomy in these patients. Peritoneoscopy was performed on 21 patients, and acute appendicitis was confirmed in 12 cases. Tubo-ovarian pathology was found in 5 patients, no pathology was found in 4 patients, and an unnecessary celiotomy was avoided in 8 patients. The incidence of normal appendix removal was reduced to 15%. There were no serious complications associated with peritoneoscopy. Diagnostic peritoneoscopy should be liberally employed in women of reproductive age with suspected appendicitis. PMID- 2972082 TI - [Work environment. Skin problems]. PMID- 2972083 TI - [Help for the home helpers]. PMID- 2972084 TI - [Proliferation and differentiation of erythroid bone marrow cells in patients with hypoplastic anemia under alpine conditions (based on 3H-thymidine incorporation data)]. PMID- 2972085 TI - [Prognostic importance of blood plasma beta-endorphin levels in the most acute period of myocardial infarct]. PMID- 2972086 TI - Corticosterone induction of cleft palate in mice dosed with orciprenaline sulfate. AB - Orciprenaline sulfate is a beta-adrenoceptor stimulant chemically described as 1 (3,5-dihydroxyphenyl)-1-hydroxy-2-isopropylaminoethane sulfate (Alupent). The drug has broncho-dilating activity and has been developed in numerous countries since 1961. The purpose of these studies was to investigate the teratogenic potential of orciprenaline and its mode of action in pregnant Jcl:ICR mice, when administered during the period of organogenesis and, more systematically, during the critical period of palate formation. Daily doses of 5, 50, and 500 mg/kg were given orally by gavage to mice on days 6-15, 11-13, or on day 12 of gestation. Additional studies were done to evaluate the maternal cardiotoxic action of orciprenaline and its effects on adrenal cortex and endogenous serum corticosterone. Five mg/kg triamcin-olone acetonide, a glucocorticoid, were given subcutaneously as a positive control causing 100% cleft palate. Myocardial necroses occurred in pregnant mice only after 500 mg/kg orciprenaline had been given, and a significant increase in cleft palate occurred if exposure took place during days 11-13 or day 12 of gestation. This increase in cleft palate can be explained by the teratogenic effect of an elevated maternal serum corticosterone level 1 hr after orciprenaline treatment, about three times the control value. PMID- 2972087 TI - Behavior of protein S during long-term oral anticoagulant therapy. AB - It has recently been reported that a natural anticoagulant, protein S (PS), is depressed during oral anticoagulation. Since more detailed information is required from the clinical standpoint, we measured plasma levels of PS [both total and free (not complexed) PS antigen], C4b-binding protein (C4bp) and other vitamin K-dependent proteins (factors II, VII, IX, X and protein C) in 60 plasma samples from patients on long-term oral anticoagulant therapy with warfarin. Together with the reduction of other vitamin K-dependent plasma proteins, PS decreased during warfarin treatment, being dependent on the intensity of the therapy. A considerable variation in plasma PS levels was also observed among individuals with a similar intensity of anticoagulation. Plasma concentration of C4bp was closely correlated with total PS level, and free PS/total PS ratio was independent of thrombotest values. These findings indicate that long-term oral anticoagulant therapy results in the suppression of the synthesis of PS, and that its reduction is on the whole balanced with C4bp and vitamin K-dependent coagulation factors. It was suggested that the metabolism of C4bp might be regulated by the plasma PS level, although this hypothesis needs further exploration. PMID- 2972088 TI - Evaluation of lupus anticoagulants: antiphospholipid antibodies, endothelium associated immunoglobulin, endothelial prostacyclin secretion, and antigenic protein S levels. AB - Plasma samples from nineteen patients with well characterized lupus anticoagulants (LA) were evaluated using a series of test systems. An ELISA was used to determine if the plasmas contained antiphospholipid antibodies (APA); fifteen of nineteen LA plasmas contained APA, with five exhibiting IgG only, two exhibiting IgM, and eight plasmas containing both IgG and IgM. Anti-phosphatidyl serine (PS) was the predominant IgG specificity and all IgM APA-containing plasmas reacted with phosphatidyl inositol (PI). An ELISA was developed to determine if LA plasmas contained immunoglobulin which would associate with cultured human umbilical cord vein-derived endothelial cells (HUV); ten of nineteen plasmas contained endothelium associated immunoglobulin (EAI). There was significant concordance between the occurrence of EAI and IgM anti-PI. The occurrence of EAI or APA, either singly or in combination, did not correlate with a past history of thrombosis. Patient plasmas were incubated with HUV and examined for effects on HUV prostacyclin (PGI2) secretion; six plasmas significantly stimulated PGI2 secretion and one plasma was inhibitory. Finally, plasma levels of free and total antigenic protein S were determined by EID. Five plasmas contained significantly reduced levels of free antigenic protein S, and total antigenic protein S was reduced in ten plasmas. Patient histories were examined for evidence of thrombotic episodes; six patients had a history of either arterial or venous thrombosis, with five of these six patients having drug induced LA. Thus, unlike previous studies, drug-induced LA were associated with thrombosis. PMID- 2972090 TI - Assessment of carbamate pesticide immunotoxicity. AB - Adult female Swiss Webster and B6C3F1 mice received distilled water only or water containing 0.1, 1.0, 10, 100 or 1000 ppb of aldicarb daily for 34 days. The target concentration of aldicarb present in the 10 ppb dosing solution was analytically verified on a daily basis as was its stability over a 48-hr period. To develop an immune profile of this compound, functional parameters measured after exposure included resistance to infectious viral challenge; quantitation of splenic antibody-forming cells to sheep erythrocytes; splenic lymphocyte blastogenesis to B-cell mitogens; and T-cell mixed lymphocyte culture response. In addition, gross and histopathologic examinations of tissues relevant to the immune system were performed. The absence of significant effects on any of these parameters suggests that aldicarb at environmentally relevant exposure concentrations is not immunotoxic in rodents. PMID- 2972089 TI - Platelet activity during acute myocardial infarction treated with tissue plasminogen activator. PMID- 2972092 TI - The origin of espundia (esponja) PMID- 2972091 TI - Theophylline prolongs survival and decreases renal damage in female NZB/W F-1 mice. AB - Theophylline (Th) augments suppressor T cell activity (STCA). We attempted to test this immunoregulatory effect of TH on survival, renal disease and several immunologic parameters in NZB/W F-1 female mice. The median survival was 274 +/- 50 days for the mice receiving Th and 235 +/- 39 days for the controls (p less than 0.01). Assessment of the extent of renal damage by light microscopy revealed activity scores of 1.86 +/- 2 and 4.71 +/- 2.7 for the Th and control groups, respectively (p less than 0.001). T cell activity of NZB/W F-1 lymphocytes was assessed by the local xenogeneic GVH reaction. The mean GVH reaction volume of the Th group was significantly lower (9.3 +/- 8.8 mm3) as compared to controls (31 +/- 9 mm3) (p less than 0.001). The proportion of Lyt-1 versus Lyt-2 spleen lymphocytes did not change significantly. It is concluded that Th prolongs survival and decreases the extent of renal damage in female NZB/W F-1 mice. PMID- 2972093 TI - Cellular immunity to Trypanosoma cruzi is mediated by helper T cells (CD4+). AB - The infection of mice with Trypanosoma cruzi has been used as an experimental model for human Chagas disease, because the murine and human infections have similar acute and chronic phases generating similar immunopathological phenomena. Histopathological studies of murine tissues showed that the inflammatory lesions were small during the acute phase and composed mainly of mononuclear cells. During the chronic phase, cellular infiltrates were clustered in large granulomata consisting of mononuclear and polynuclear neutrophil cells. Characterization of the infiltrating cells by surface markers showed that about 6% were Thyl.2+ T cells, and CD4+(Lyt 1+) T cells (T helper/DTH subset) were more numerous than CD8+(Lyt 2+) T cells. These observations suggest that delayed type hypersensitivity plays a role in the pathology of Chagas disease. PMID- 2972095 TI - Pretreatment of chronic myelogenous leukemic blood samples with cytosine arabinoside prior to mixed lymphocyte culture typing. PMID- 2972094 TI - In vivo fluorescence microscopy of kidney subcapsular blood flow in mice. Effects of cyclosporine, (NVA2)-cyclosporine, and isradipine, a new calcium antagonist. AB - The subcapsular kidney microcirculation in mice was observed through a fluorescence microscope, recorded on videotape, and examined for response to infusions of cyclosporine A (CsA) and cyclosporine G (CsG). Coded video recordings were evaluated by a semiquantitative method. CsA infusion (1.6 +/- 0.4 mg/kg/min) induced a nearly complete inhibition of the subcapsular blood flow. At lower infusion rates (0.46 +/- 0.2 mg/kg/min), the blood flow inhibition was less pronounced. CsG infusions at corresponding rates induced significantly less inhibition. Pretreatment with a new calcium antagonist, isradipine (18-20 micrograms/kg bwt), completely prevented the CsA-induced impairment of subcapsular microcirculation. The calcium antagonist, however, did not improve blood flow when administered after induction of inhibition by CsA (16.8 +/- 2.5 mg/kg), emphasizing the importance of pretreatment. This study suggests hypoperfusion due to vasoconstriction as an important pathophysiologic mechanism for CsA-induced nephrotoxicity. CsG, when given at corresponding rates, induced less inhibition of the blood flow. Pretreatment with a calcium antagonist, isradipine, completely prevented a CsA-induced inhibition of blood flow, suggesting a potential value in the prevention of CsA-induced nephrotoxicity. PMID- 2972096 TI - Resolution of streptozotocin-induced diabetes mellitus by pancreatic transplantation in inbred mice. PMID- 2972097 TI - [Monoclonal antibody metabolism during the growth of hybridoma cells]. AB - Metabolism of monoclonal antibodies (MAb) during the growth of mouse hybridoma, producing MAb to phage lambda, has been studied. It was shown that the specific production of MAb decreased by 25-35% in the stationary phase of growth in comparison with the middle of the exponential growth phase, which was associated with the decrease in the rate of MAb synthesis. The secretion kinetics of MAb did not change during the growth of hybridoma cells. MAb did not degrade inside the cells and in the culture medium after being secreted. The ratio of the synthesis rate of MAb to that of cellular proteins increased from 7-10% in the exponential growth phase to 14-18% in the stationary phase, which points to a specific regulation of MAb synthesis in comparison with cellular proteins. Possible regulation mechanisms for synthesis of MAb and cellular proteins during the growth of hybridoma cells are discussed. PMID- 2972098 TI - [Inhibition of the activity of membrane-bound Ca2+-ATPase in the sarcoplasmic reticulum fragments of rabbit skeletal muscles by the alkaloid sanguinarine]. AB - Sanguinarine, a plant DNA-intercalator, is shown to inhibit the enzyme activity of the membrane-bound Ca2+-ATPase of rabbit skeletal muscle sarcoplasmic reticulum fragments. This inhibition could be interpreted by the well known ability of this alkaloid to interact with sulphydryl groups of the enzymes. Sanguinarine is a weaker inhibitor of this reaction than a sulphydryl group poison Ag+. The I50 is 3.10(-6) M for Ag+ and 7.10(-5) M for sanguinarine in the reaction medium with NO3- substituted for Cl-. In the standard reaction medium containing Cl-, the I50 for sanguinarine is 1.8.10(-4) M. In this case sanguinarine activates Ca2+-ATPase at low concentrations presumably because of uncoupling ATP hydrolysis from Ca2+ transport through membrane. Other agents studied are: DNA-intercalators--ethidium bromide, acriflavine, acridine orange; DNA-complexing antibiotics--actinomycin D, and olivomycin, alkaloids, quinine, morphine, berberine and an uncoupler of oxidative phosphorylation 2,4 dinitrophenol. These were found not to inhibit Ca2+-ATPase activity up to the concentrations of 10(-3)-10(-4) M. PMID- 2972100 TI - Conventional and color Doppler in the assessment of fetal and maternal circulation. PMID- 2972099 TI - [Sister chromatid exchanges, during x-irradiation, in the blood lymphocytes of patients with hereditary diseases and radioresistant DNA synthesis]. AB - X-ray irradiation induced sister chromatid exchanges (SCE) in blood lymphocytes from patient with Down's syndrome and adult progeria (in both the cases radioresistant DNA synthesis takes place). In these diseases, likely as upon form II of xeroderma pigmentosum (the replicative DNA synthesis is radioresistant), X ray irradiation lowers the rate of SCE compared with that in the control, then the SCE rate rises with the increase in radiation dose, reaching the rate of SCE in non-irradiated cells. In normal lymphocytes (in which ionizing radiation inhibits the replicative synthesis of DNA) the rate of SCE rises with the rise of radiation dose. Thus, the rate of SCE in X-ray irradiated lymphocytes is in reverse dependence with radioresistance of replicative synthesis of DNA. The data obtained are explained in accordance with the replicative hypothesis of the SCE nature (Painter, 1980a): in cells of patients with Down's syndrome, xeroderma pigmentosum form II and progeria of adults the time of existence of partly replicated clusters of replicons is decreased due to radioresistant replicative synthesis of DNA, but the presence of partly replicated clusters of replicons is necessary for SCE formation. Therefore the rate of SCE in X-irradiated cells of these patients decreases. PMID- 2972101 TI - Urodynamics of the lower urinary tract. AB - The principles and techniques of uroflowmetry, cystometry, pressure-flow studies and urethral pressure profilometry are reviewed. The interpretation of urodynamic tests depends on the methodology used and whether the patients usual voiding symptoms during the examination. The more complicated urodynamic tests may require computer assistance both for data storage but also for test interpretation. One of the main challenge in future urodynamic is to transform these tests to clinical usable tools. PMID- 2972103 TI - [Current views of prune belly syndrome]. AB - The recent literature is reviewed and the etiology, pathogenesis, diagnosis, therapy and prognosis of prune belly syndrome are presented and illustrated with reference to personal experience in typical cases. It seem possible that the syndrome is not an entity in its own right, since it also appears in incomplete forms. Prenatal sonography allows an early diagnosis, but to date intrauterine treatment is not indicated. Postnatal operative intervention in the form of major reconstructive surgery has to be viewed very critically. So far, no positive effects on long-term prognosis have been proven, so that, depending upon the severity of the syndrome, treatment should be as conservative as possible. PMID- 2972102 TI - Effects of long term GnRH analogue treatment on hormone levels and spermatogenesis in patients with carcinoma of the prostate. AB - The effects of long term GnRH treatment with the biodegradable depot formulation of ICI 118.630 on hormone levels and spermatogenesis were investigated in 18 males with advanced prostate cancer. Plasma levels of FSH, LH, testosterone, DHEA S and SHBG were monitored at regular intervals. The drug suppressed FSH, LH and testosterone significantly and did not affect DHEA-S and SHBG plasma levels. Tissue specimens for histologic assessment and quantitative analysis of germinal cell types were obtained at secondary orchidectomy in 16 patients immediately following GnRH analogue treatment. Germinal cell maturation was arrested at the spermatogonial stage. In two patients discontinuing treatment histologic assessment of secondary orchidectomy specimens 9 and 10 months after the last GnRH analogue depot injection resulted in germinal cell maturation to late spermatids in part of the tubule cross sections. These results indicate that long term administration of the GnRH analogue fails to produce complete testicular sclerosis and spermatogenic arrest might be reversible. PMID- 2972104 TI - Effect of long-acting LHRH analog (Zoladex) on prostate cancer: evaluated by transrectal ultrasonography. AB - Transrectal ultrasonography has proved valuable in assessing the effect of primary treatment modalities for prostate carcinoma. This study shows patients who had a significant reduction in primary tumor volume had a significantly better prognosis and had less local symptoms than did the group of patients that did not have a significant reduction (less than 50%) in primary volume secondary to therapy. Patients were treated with either castration or Zoladex and all had Stage D2 cancer of the prostate. PMID- 2972105 TI - Management of cloacal exstrophy. AB - This case study demonstrates our experience in the very complex management of cloacal exstrophy. It exemplifies the aggressive therapy by a dedicated multidisciplinary health team and the excellent postoperative results. PMID- 2972106 TI - [The staged team method in the ophthalmological care of children]. PMID- 2972107 TI - A comparison of the efficacy of four different long-acting boluses to prevent infections with Dictyocaulus viviparus in calves. AB - A field study of calves in their first grazing season tested the efficacy of four long-acting devices--a morantel sustained-release bolus, a levamisole sustained release bolus, an oxfendazole interval bolus, and an albendazole interval bolus- against Dictyocaulus viviparus. The pasture had been previously contaminated by four calves orally inoculated with infective lungworm larvae. The calves were grazed together with four bolus-treated groups, each comprising four calves. Lungworm infection became patent in the experimentally inoculated calves between 22 and 26 days. Infection in the bolus-treated groups became patent after 54 days. The morantel bolus group excreted the most larvae, followed by the albendazole bolus group, and the levamisole bolus group. The oxfendazole bolus group excreted by far the least larvae. Eosinophil curves and ELISA titres showed that treated groups had essentially the same course of infection. The heavy infection to which the treated calves were exposed produced complete immunity in all groups. Challenge infection of 10,000 larvae at housing did not change any of the test parameters. Post-mortem examination showed only one positive calf with few worms. We concluded that when pastures are heavily infested with lungworm larvae, all boluses prevent severe clinical signs and allow build up of solid immunity, although none completely prevent excretion of larvae. PMID- 2972108 TI - Therapeutic efficacy of medicating drinking water with spectinomycin and lincomycin-spectinomycin in experimental Escherichia coli infection in poultry. AB - Although no microbiological activity was found in blood plasma during treatment of broilers through the drinking water with spectinomycin or lincomycin spectinomycin, these drugs proved highly effective in treating experimentally induced colibacillosis. This efficacy may be produced by a metabolite or a degradation product of spectinomycin that reaches the infection site in the respiratory tract and interferes with adherence of bacteria. PMID- 2972109 TI - Epidemiological patterns of gastrointestinal and lung helminth infections in grazing calves in The Netherlands. AB - The patterns of gastrointestinal and lung helminth infections in grazing calves were studied in two small grazing experiments carried out between May and October in 1985 and 1986, respectively, at Utrecht State University. For lungworm a very rapid translation of infective larvae was seen in both experiments, resulting in a rapid build-up of the worm burdens, and in July the second generation of Dictyocaulus caused husk. After the end of patency for lungworm, in the beginning of August, a rapid decrease in pasture infectivity, measured by pasture larval counts and worm counts of tracer calves, was observed. For the gastrointestinal nematodes the translation of infective larvae was much slower as a distinct 'midsummer increase' of pasture infectivity was not seen before the end of July. The worm counts of the tracer calves and the pasture larval counts showed that these larvae remained on pasture much longer than the lungworm larvae. As in Dictyocaulus, patency stopped in August-September in Nematodirus but not in Ostertagia and Cooperia, indicating a slower rate of development of immunity in both latter genera. In contrast to lungworm, the results indicate a clear and predictable seasonal pattern for gastrointestinal helminth infections. PMID- 2972110 TI - The effect of halofuginone on the excretion of Salmonella typhimurium by experimentally infected chickens. AB - The effect of feeding halofuginone at 3 and 6 ppm on the excretion of Salmonella typhimurium by experimentally infected chickens was studied. A standardized procedure was used involving the oral inoculation of 72-h-old specific pathogen free chickens with 10(8) organisms of a nalidixic acid-resistant mutant of a strain of S. typhimurium. At weekly intervals, cloacal swabs were taken and a semi-quantitative assessment was made of the numbers of Salmonella organisms excreted. When compared with the control group of infected chickens fed unmedicated food, the group fed halofuginone at 3 ppm showed no significant increase in excretion rate. The group fed 6 ppm showed a slight increase in excretion which was statistically significant. PMID- 2972112 TI - Packaging and transduction of non-T3 DNA by bacteriophage T3. AB - A defined in vitro system for packaging T3 DNA also packaged other linear DNAs, including T4, lambda, and plasmid DNAs. The packaging capacity was determined to be 40 kb (kilobase pairs) by measuring the packaged length of T4 DNA. Packaged lambda and plasmid DNAs were injected into host cells to form plaques and transductants, respectively. The yield of transducers increased by using artificially ligated plasmid oligomers. The T3 mutant in gene 3 endonuclease (T3 3-) packaged plasmid DNA during abortive infection and transduced it into the recipient. Transduction of recombinant plasmids was not affected by the presence of the terminally redundant sequence (TR sequence) but increased by 4 orders of magnitudes when the genetic right-end 2.7-kb sequences, containing gene 19 (E1) but lacking TR, were present and by 7 orders when both E1 and TR sequences were present. However, these sequences did not increase transduction of these plasmids by T7 3-. Analysis of the structure of transduced plasmid DNAs indicates that transducing particles carry head-to-tail oligomers of plasmid DNA with the same termini as those of T3 genomic DNA. The mechanism of formation of transducing particles is discussed. PMID- 2972111 TI - Reactivity of eleven anti-human leucocyte monoclonal antibodies with lymphocytes from several domestic animals. AB - Nine commercially available monoclonal antibodies and two monoclonal antibodies from The American Type Culture Collection, raised against various human leucocyte surface antigens, were tested on lymphocytes from cow, sheep, goat, swine, horse, cat, dog, mink, and rabbit as well as man. Four antibodies bound to lymphocytes from some of the animals. These were the antibodies against CD8 and CD4 antigen, the antibody to C3b-receptor, and the antibody to the HLA-DR antigen. The CD8 antigen-reactive antibody reacted with lymphocytes from mink, cat, dog, and sheep, while the CD4 antigen-reactive antibody reacted with lymphocytes from mink. The anti-C3b-R antibody reacted with lymphocytes from horse, swine, dog, and cat, and the anti-HLA-DR reacted with lymphocytes from cow, goat, sheep, horse, dog, cat, and mink. PMID- 2972113 TI - Analysis of lettuce necrotic yellows virus structural proteins with monoclonal antibodies and concanavalin A. AB - Three major structural proteins of lettuce necrotic yellow virus (LNYV) were identified by discontinuous polyacrylamide gel electrophoresis (PAGE) to have Mr approximately 78,000 (G), 57,000 (N), and 19,000 (M). Unreduced G and M proteins had faster mobilities in PAGE indicating the presence of disulfide bonds. The G protein was shown to be glycosylated with a complex network of oligosaccharides containing beta-N-acetylchitobiose N-linked to asparagine residues of the protein. Up to 17 additional minor bands were also detected in silver-stained electrophoretograms. In Western immunoblots, 9 of these (Mr approximately 27,000 220,000) were recognized by a monoclonal antibody to the N protein and another 6 (Mr approximately 58,000-180,000) with a monoclonal antibody to the G protein, indicating that they were degradation products or aggregates of these two viral proteins. Two minor silver-stained bands failed to react with either of the monoclonal antibodies, but were recognized by polyclonal anti-LNYV serum and are probably the L (Mr approximately 190,000) and NS (Mr approximately 38,000) viral proteins. PMID- 2972114 TI - Mechanism of interferon action. Expression of reovirus S3 gene in transfected COS cells and subsequent inhibition at the level of protein synthesis by type I but not by type II interferon. AB - The effect of interferon on the expression of the reovirus serotype 1 Lang strain S3 gene was examined in simian COS cells transfected with the expression vector pSVS3 containing the S3 gene under the control of the SV40 late promoter. When COS cells were treated with type I interferon-alpha 24 hr after transfection, the synthesis of the reovirus S3-encoded sigma NS polypeptide was inhibited about 10 fold as compared to that in untreated control cells. By contrast, under the same conditions, neither the plasmid DNA copy number nor the S3 gene mRNA levels were significantly affected by interferon treatment. Type II interferon-gamma, unlike the type I interferons-alpha, did not affect the rate of synthesis of polypeptide sigma NS in pSVS3-transfected cells. PMID- 2972115 TI - Characterization of a unique protein produced by influenza A virus recovered from a long-term persistent infection. AB - Virus isolated from a persistent infection initiated in BHK cells with influenza A/WSN/33 (H1N1) produced an unusual pattern of protein synthesis in productive infections of BHK cells: The levels of NP and M1 proteins were slightly reduced compared to an infection with wild-type WSN, while the other proteins (Pb1, Pb2, Pa, HA, NS1, and NS2) were synthesized at very low or undetectable levels. In addition, a new viral protein with an approximate molecular weight of 11 kDa (Pi protein) is made (Frielle et al., Virology 138, 103-117, 1984). When viral RNA was analyzed by the Northern blot technique, a deletion was found in the NS gene segment and in NS1 mRNA; all other RNAs were full-sized. Immunoprecipitation of in vitro translation products demonstrated that the Pi protein reacts specifically with anti-NS1 serum. In addition, the Pi protein, like the NS1 of the parental wild-type virus, accumulated in the nucleus of infected cells. These results indicate that the Pi protein is a mutated form of the NS1 protein encoded by a deleted NS segment and suggest that this mutation may be involved in the expression of the persistent virus phenotype. PMID- 2972116 TI - [Incidence of ECG changes in the relatives of patients with hypertrophic cardiomyopathy without concurrent myocardial hypertrophy]. PMID- 2972117 TI - [Medical support of the Soviet troops in the Kursk defensive operation]. PMID- 2972119 TI - [Occupational rehabilitation--a responsibility in managing the industrial health system]. PMID- 2972118 TI - [Newly isolated strains of the Batken virus]. PMID- 2972120 TI - [International classification of impairments, disabilities and handicaps]. PMID- 2972121 TI - [Professional employment of graduates of rehabilitation centers for occupational education in East Germany l973-l985]. PMID- 2972122 TI - [Management of handicapped children and adolescents in Berlin-Marzahn child and adolescent protective service]. PMID- 2972123 TI - [HLA-associated predisposition to disease and mechanisms of its realization]. PMID- 2972124 TI - [Phytodermatoses]. PMID- 2972125 TI - [Blood T-helper and T-suppressor count of patients with gonorrheal orchido epididymitis]. PMID- 2972126 TI - [Psoriasis morbidity in Aktyubinsk Province, Kazakh SSR]. PMID- 2972127 TI - [DDS in the treatment of necrotic acne]. PMID- 2972128 TI - [A rhinophyma-like form of squamous cell skin cancer]. PMID- 2972129 TI - [Treatment of dermatophytoses and onychodystrophies using metronidazole ointment]. PMID- 2972131 TI - [The significance and meaning of genetic and environmental factors for the etiology and pathogenesis of dysgnathias]. PMID- 2972130 TI - Immunocytochemical study on the ultrastructural localization of human-type ABO (H)-blood group activities in a macaque (Macaca irus). AB - The immunocytochemical study on the ultrastructural localization of human-type ABO(H)-activities in a crab-eating macaque (Macaca irus) was carried out by using postembedding and immuno-gold staining method. The tissue specimens examined were the esophagus, stomach (St), small intestine (Si), large intestine, liver, kidney, and pancreas. The specimens from these organs and submandibular gland (Sg) of a human (O-group) were used as staining reaction controls. Primary and secondary antibodies were commercially obtained mouse monoclonal anti-A, -B, -H (IgM), and goat anti-mouse IgM labeled with colloidal gold particles (luminal diameter 20 nm), respectively. The results were as follows: (1) In macaque specimens, only A-activity could be observed as the location of gold particles on the peripheral rim of serous secretory granules (Sg) and of epithelial cells (esophagus), the mucous droplets in epithelial cells and brush border (St, Si), the intracellular secretory canaliculi [ISC (St)] and the zymogen granules and secretory ducts (pancreas). Gold particles could be also noted at the Golgi apparatus and nascent secretory granules. (2) By periodic acid-thiocarbohydrazide osmium tetroxide (PA-TCH-OS) reaction, hexose-rich neutral mucopolysaccharides were noted on the peripheral rim of serous secretory granules (Sg), the mucous droplets (St, Si), the ISC (St), and the brush border (Si). Such a distributional pattern corresponded well with that of gold particles, indicating that the substances were responsible for ABO(H)-activities. PMID- 2972132 TI - [Development of intermaxillary comparison values in the dysgnathic bite]. PMID- 2972133 TI - [The growth pattern of patients with cleft lip and palate]. PMID- 2972135 TI - [Dental research on patients with anhidrotic ectodermal dysplasia (AED)]. PMID- 2972134 TI - [Dentition parameters and correlations--results, considerations, conclusions]. PMID- 2972136 TI - [In vitro studies of caries-induced dentin transparency]. PMID- 2972138 TI - [Our current concept of preprosthetic surgery]. PMID- 2972137 TI - [Comparative biological and microbiological testing of root canal filling materials]. PMID- 2972139 TI - [Do carbohydrate compounds represent the initial plaque matrix?]. PMID- 2972140 TI - [A method for the combined electron microscopic study of sputter films]. PMID- 2972141 TI - [New progestational hormones]. AB - The pharmacology and the application in oral hormonal contraceptives of four new progestogens--gestodene, desogestrel, norgestimate, and dienogest--are described in a survey. Although derived from 19-norethisterone these progestogens exhibit in clinical dosages no androgenic effects. Both in animal and in human tests gestodene possesses the highest progestogenic potency followed by levonorgestrel, desogestrel, norgestimate, and dienogest. The monophasic preparations with the new progestogens (femovan--gestodene, marvelon--desogestrel, cilest- norgestimate, certostat--dienogest) are safe contraceptives and the incidence of bleeding disturbancies amount to about 5%. After administration of these monophasic contraceptives the concentration of SHBG in serum increases and as a result of this the level of free testosterone decreases. Further a favourable effect on lipid metabolism is observed. In spite of a different progestogen content these combination pills are almost without influence on biochemical parameters and carbohydrate metabolism. PMID- 2972142 TI - [Artashes Ivanovich Karamian (on his 80th birthday)]. PMID- 2972143 TI - [Biochemical mechanisms of temperature adaptation in the chick embryo]. AB - The effect of a short-term cooling of the incubated eggs has been investigated on the intensity of oxidative phosphorylation and the activity of ATPase in mitochondria from muscles and liver of chick embryos and chicks. It was found that the decrease of temperature increases oxygen consumption in muscle mitochondria decreasing esterification of inorganic phosphate. As a consequence, the value of P/O decreases. The activity of ATPase significantly increases. Uncoupling between oxidation and phosphorylation in liver mitochondria takes place more slowly. It is suggested that these changes account for realization of thermoregulation. PMID- 2972144 TI - [Virulence of Streptococcus pneumoniae strains belonging to different serovars]. AB - The study of the virulence of 352 S. pneumoniae strains isolated from patients with inflammatory lung diseases revealed that their virulence depended, to a certain extent, on the state of the polysaccharide capsule of streptococci, as among 299 typed cultures 38.1% were virulent, while out of 53 nontypable strains of these bacteria only 8 strains (15.1%) proved to be highly pathogenic for white mice and all S. pneumoniae R-forms proved to be avirulent. All 11 S. pneumoniae strains under study belonging to serovars 1 and 2 and 87% of the cultures belonging to serovar 3, isolated from patients with inflammatory lung diseases in Leningrad, were highly virulent. The characteristic feature of S. pneumoniae cultures of other serotypes was their wide spectrum of pathogenicity. S. pneumoniae cultures isolated from the spinal fluid of patients with pneumococcal meningitis also differed in their pathogenicity levels but strains highly pathogenic for mice prevailed. PMID- 2972145 TI - [Methodological bases for the study and the patterns of the seasonality of viral hepatitis A]. AB - On the basis of the study of the data on the incidence of viral hepatitis A in one of the districts of a big city for 20 years the authors come to the conclusion on the advisability of a comparative study of the monthly changes in the incidence of hepatitis A within individual uniform morbidity cycles covering the periods of many years and the use of average monthly data for many years for plotting the typical curve, as well as the use of the simplified for the calculation of the upper limit of annual morbidity. Similarities and differences in the monthly dynamics of morbidity in the years of high and low morbidity levels have been revealed, and the age group of the population (20-39 years) which ensures the continuity of the epidemic process all the year round has been determined. The factors contributing to the seasonal activation of the epidemic process start operating in June among schoolchildren aged 11-14 years, and later their operation spreads to other groups of the population. PMID- 2972146 TI - [Multiyear and seasonal hepatitis A morbidity in different population age groups in certain areas]. AB - At the areas with high activity of hepatitis A (HA) epidemic process the duration of epidemic cycles was shown to differ, the intervals between the peaks of morbidity increasing in older age groups. The beginning of seasonal rises exceeding the average annual HA morbidity level in different age groups was found to depend on the activity of the epidemic process. At the areas with the highest activity of the epidemic process children aged 1-2 years were the first to be affected by the seasonal rise of HA. Stable direct correlation between HA morbidity levels at the beginning of seasonal rises and some markers indicative of unfavorable sanitary conditions (the size of the fly population, the purity of water samples deviating from the requirement of the Government Standard) during the preceding year was demonstrated. PMID- 2972147 TI - Up-to-date surgical management of tubal infertility due to inflammations of the pelvis. AB - A total of 87 refertilization operations in the distal tubal segment are reported. Tubal obstructions developed following inflammations of the pelvis. Hysterosalpingography and laparoscopy were used to learn about the patients' anatomy and these served as bases for assessing the changes of pregnancy following recanalization. The interventions were made by applying up-to-date microtechnical methods and instruments. Among the causes of tubal infertility, the interruption of the first pregnancy showed a marked prevalence. The 20 pregnancies that ensued correspond to a success rate of 23%. Mature deliveries occurred in 17 cases. A considerable proportion of the pregnancies occurred within a year of the operation. The effectiveness is mainly influenced by adhesions in the pelvis and the characteristics of the distal tubal segment. The surgical solution is worth attempting when the expected chances are over 20%. PMID- 2972148 TI - Local treatment of venous leg ulcers. AB - At present the following guidelines for treatment of venous leg ulcers from our department are: Check the patients' general health--special attention to heart incompensation with oedema of the legs and the peripheral circulation. The most relevant laboratory tests are haemoglobin and urine-glucose. Routine bacterial cultivation is not necessary in non-diabetic patients as the result will generally not influence diagnosis, treatment or prognosis. Furthermore, treatment with topical antibiotics should be avoided. It is not only unnecessary, wasteful and sensitizing, but it also involves a risk of causing antibiotic resistance. Systematic antibiotic therapy is indicated only when obvious inflammatory signs in the tissues surrounding the ulcer are present, e.g. erysipelas or cellulitis. Non-sensitizing topical remedies should be applied. Avoid wool, alcohols, parabens, topical antibiotics and oxiquinolines. When eczema occurs use a hydrocortisone preparation in an inert base. Epicutaneous testing might be indicated. Bandages to be recommended are double-layer bandages consisting of an inner zinc oxide impregnated stocking and an outer elastic bandage and hydrocolloid dressing plus compression bandage. PMID- 2972149 TI - Treatment of venous leg ulcers with cadexomer iodine with particular reference to iodine sensitivity. PMID- 2972150 TI - Hormonal responses to synthetic atrial natriuretic peptide in patients on regular hemodialysis. AB - The effects of atrial natriuretic peptide (ANP) on mean arterial blood pressure, heart rate, plasma renin activity, aldosterone, cortisol, norepinephrine, epinephrine and arginine vasopressin were studied in 6 anuric subjects receiving regular hemodialysis. An iv bolus injection of 8 nmol of ANP followed by infusion at 32 pmol.kg-1.min-1 for 1 h in the pre- and posthemodialysis period was performed. Basal plasma ANP was higher before than after hemodialysis. ANP administration produced a reduction in mean arterial blood pressure accompanied by an elevation of norepinephrine and of plasma renin activity (from 2.49 +/- 0.52 to 3.39 +/- 0.85 nmol.1-1.h-1 predialysis and from 2.78 +/- 0.71 to 3.15 +/- 0.86 nmol.l-1.h-1 postdialysis, respectively, mean +/- SEM; P less than 0.05). Plasma aldosterone and cortisol were significantly decreased. Plasma epinephrine and AVP remained unchanged. These hemodynamic and hormonal changes were similar in the pre- and the postdialysis period. These results suggest that 1) ANP causes a fall in mean arterial blood pressure, which in turn induces reflex tachycardia and activation of the sympathetic nervous system without diuresis; 2) the activated sympathetic nervous system as reflected in elevation of plasma norepinephrine may increase plasma renin activity; 3) reduced plasma aldosterone is not influenced by enhancement of the renin-angiotensin system; therefore, 4) reduction of plasma aldosterone as well as cortisol is probably due to direct action of ANP, and finally 5) AVP had no direct relation with ANP administration. PMID- 2972151 TI - Prolonged influence of a neonatal cyproterone acetate treatment on renal androgen binding in mice. AB - To determine whether neonatal endogenous androgens influence adult renal androgen binding, newborn male mice were injected from 1 to 10 days of age with cyproterone acetate and newborn females with testosterone from 1 to 10 days and from 20 to 40 days of age. In controls, at adulthood, the total cellular androgen receptor content was significantly higher in males (1700 +/- 200 receptors per cell) than in females (1060 +/- 50) and, as expected, the nuclear receptor content was 12-fold higher in males. While the total number of receptors (1650 +/ 200 per cell) was unchanged in adult males neonatally treated with cyproterone acetate, their distribution between cytosol and nucleus was similar to that in control females despite normal circulating and renal testosterone levels. The nuclear receptors represented 50, 7 and 11% of the total receptors in control males, control females and cyproterone acetate-treated males, respectively. The very low levels of nuclear receptors present in the kidney of cyproterone acetate treated males probably explain the decreased sensitivity of this organ to testosterone. The nuclear receptor accumulation measured in adult animals after a single injection of testosterone did not seem to be affected by neonatal hormonal manipulations. PMID- 2972152 TI - Effects of an oral water load and intravenous administration of isotonic glucose, hypertonic saline, mannitol and furosemide on the release of atrial natriuretic peptide in men. AB - The effects of an oral water load and iv administration of isotonic glucose, hypertonic saline, mannitol and furosemide on release of human atrial natriuretic peptide (hANP) were examined in normal subjects to determine the main factors causing its release. In addition, the influence of age on hANP secretion was investigated. The mean plasma hANP level in normal subjects, 0-89 years old, was 20.6 +/- 1.1 ng/l (mean +/- SEM) and showed age-related change. The plasma hANP level did not change significantly after a water load or infusion of isotonic glucose, but rose significantly from 11.4 +/- 1.4 to 15.6 +/- 3.2 ng/l after infusion of hypertonic saline and from 10.9 +/- 1.6 to 17.8 +/- 4.1 ng/l after infusion of 20% mannitol in parallel with the increase in plasma volume. The plasma hANP level decreased from 17.3 +/- 2.5 to 9.0 +/- 2.5 ng/l after injection of 40 mg of furosemide. A positive correlation was found between change in the plasma hANP level and percent change in the plasma volume (P less than 0.001) on these treatments. The response of plasma hANP to hypertonic saline infusion was greater in older than in young men. These results indicate that 1) the secretion of hANP shows an age-related change and 2) increase in the circulating plasma volume is an important factor regulating hANP secretion. PMID- 2972153 TI - [Does the atrial natriuretic factor play a role in cirrhosis?]. PMID- 2972154 TI - Hemodynamic changes during laparoscopy with positive end-expiratory pressure ventilation. AB - Hemodynamic measurements were performed in 10 healthy women undergoing elective laparoscopy for the investigation of infertility. A standardized anesthetic technique which included the application of positive end-expiratory pressure (PEEP), 0.49 kPa (3.7 mmHg) was utilized. The following variables were studied: cardiac output, stroke volume and left ventricular ejection time (determined non invasively with impedance cardiography), heart rate, blood pressure, total peripheral vascular resistance and end-tidal carbon dioxide (ET-CO2). The combination of 25 degrees head-down tilt and PEEP ventilation during laparoscopy was associated with a pressure response that restored arterial pressures to essentially pre-anesthetic levels. Net cardiac effects were small. With this regime low pressure 0.7-1.1 kPa (5-8 mmHg) intra-abdominal insufflation with CO2 was associated with only minor cardiovascular changes. There were no indications that 0.49 kPa PEEP during laparoscopy produced adverse cardiovascular effects. The application of PEEP reduced (P less than 0.001) ET-CO2. There was no net increase in ET-CO2 after CO2-insufflation compared to the measurement after induction of anesthesia. This is in contrast to earlier studies without PEEP where a significant net increase in ET-CO2 was reported after CO2-insufflation. PMID- 2972155 TI - Evaluation of motor blockade by isometric force measurement and electromyographic recording during epidural anaesthesia--a methodological study. AB - Methods for assessing motor blockade by means of isometric force measurements and surface electromyographic (EMG) recordings in the lower extremities and abdominal wall were evaluated in 30 volunteers. The coefficients of variation were 10% for force measurements and 14% and 20% for average rectified EMG (RIEMG) recordings over the quadriceps muscle and abdominal muscles, respectively, and 8% overall for TURNS (the number of changes in the sign of the direction of the EMG signal). Seven of the 30 volunteers received epidural anaesthesia with 20 ml of mepivacaine 2% with adrenaline. The mean maximal cephalad analgesic level was T9. At that abdominal segment, RIEMG showed a reduction of 50% and TURNS of 20%. Isometric force and RIEMG recorded simultaneously in the quadriceps muscles during epidural anaesthesia displayed a linear relationship with a correlation coefficient of 0.91. TURNS was insensitive to force variations above 60% of maximum voluntary contraction. During the regression phase, 90% of both the initial force and RIEMG value was noted 180 min after the epidural injection. It is concluded that recording of RIEMG is a good method for quantitative assessment of motor blockade during epidural anaesthesia. PMID- 2972156 TI - Anesthetic management of the hypertensive patient. PMID- 2972157 TI - Congenital fiber type disproportion in an adult: a morphometric and microchemical study. AB - A 20-year-old man with marfanoid habitus had a history of congenital hypotonia and muscle weakness. Muscle biopsy showed extreme fiber type disproportion. There was total absence of Type 2B fibers. The severely hypertrophic Type 2A fibers showed twice the normal concentration of creatine phosphate at rest. These advanced morphometric, histochemical and biochemical changes may be interpreted as compensatory phenomena, which may explain the patient's pronounced functional improvement with advancing age. PMID- 2972158 TI - Comparative trial of the effects on glucose tolerance and lipoprotein metabolism of two new oral contraceptives containing gestoden and desogestrel. AB - The effects of two new low-dose oral contraceptives (triphasic ethinyl estradiol/gestoden and monophasic ethinyl estradiol/desogestrel) on glucose tolerance, plasma insulin response to glucose, fasting plasma cortisol, triglycerides (TG), total cholesterol (C), HDL-C, LDL-C, VLDL-C and sex hormone binding globulin (SHBG) were investigated in two groups of healthy women (n = 10). Investigations were performed prior to hormone ingestion and after treatment for 2 and 6 months. In both groups, fasting plasma levels of glucose and insulin as well as the areas below the glucose concentration curves were unchanged during treatment, whereas the insulin response to oral glucose was equally increased (p less than 0.05). Intake of both compounds was followed by similar increases in the levels of HDL-C (p less than 0.05) and in the HDL-C/total-C (p less than 0.05). A transient decrease in the levels of LDL-C was observed in both groups after two months. During intake of the gestoden-containing compound increases in VLDL-C and TG levels were registered after six months (p less than 0.05). Plasma levels of SHBG increased similarly in both groups (p less than 0.01). The study indicates, that intake of both hormonal compounds is free from adverse effects on glucose tolerance and lipoprotein metabolism known to be of clinical significance. No differences in the metabolic effects were found between the two compounds. PMID- 2972159 TI - Ethinylestradiol and desogestrel, alone or in combination with two doses of estriol. Effects on plasma proteins. AB - Nineteen fertile women were treated with three differently composed oral contraceptives in a cross-over study and were randomly allocated to begin with one of the three preparations. The tablets contained 0.030 mg ethinylestradiol +0.15 mg desogestrel alone (A) or in combination with either 0.5 mg estriol (B) or 2 mg estriol (C). After two treatment cycles on each preparation, blood samples were obtained and analysed for ceruloplasmin, SHBG, prealbumin, transcortin and thyroxine-binding globulin. No differences were found in the induction of liver protein synthesis. It was concluded that the addition of estriol in doses of 0.5 or 2 mg did not influence the effects induced by ethinylestradiol. PMID- 2972160 TI - Multicenter trial of a monophasic oral contraceptive containing ethinyl estradiol and desogestrel. AB - A clinical trial was conducted at 47 centers in 11 countries to assess the efficacy and acceptability of a monophasic oral contraceptive containing 30 micrograms ethinyl estradiol and 150 micrograms desogestrel. 1,613 women took part for a total of 23,258 cycles. One pregnancy occurred in a cycle where two consecutive tablets had been forgotten. Cycle control was excellent, with reported decreases in the duration and amount of withdrawal bleeding during consecutive treatment cycles and a low incidence of irregular bleeding. Blood pressure was not affected during 2 years of use. The incidence of minor side effects was already low in the first treatment cycle and decreased further in the subsequent cycles. The combination (Marvelon) was shown to be a very reliable and acceptable oral contraceptive. PMID- 2972161 TI - Oocyte retrieval in an IVF program. A comparison of laparoscopic and transvaginal ultrasound-guided follicular puncture. AB - From 1 April 1986 to 20 May 1987, 160 oocyte retrievals were performed in our IVF program. 80 oocyte retrievals were done by laparoscopy under general anaesthesia and 80 under light sedation and local analgesia using transvaginal ultrasound guided follicle puncture. More oocytes were recovered with the ultrasound procedure than with laparoscopy, 616 versus 478. The fertilization rate was higher when the oocytes were collected with the ultrasound procedure, vis-a-vis laparoscopy, 70.6% versus 59.6%. The number of embryos transferred was greater in the former than in the latter group, 300 versus 182. There were 13 clinical pregnancies in the laparoscopy group versus 17 in the ultrasound group. There were no serious complications in either group. Transvaginal ultrasound guided follicle aspiration is a safe and simple procedure. PMID- 2972162 TI - Fat-soluble 17 beta-estradiol: a way of reducing dosage in steroid hormonal substitution? AB - Eight ovariectomized women were given 0.5 mg 17 beta-estradiol cyclo-octyl acetate (E2COA) dissolved in arachis oil + 0.15 mg desogestrel, and 2 mg micronized 17 beta-estradiol (mE2) + 0.15 mg desogestrel orally in a crossover fashion for 20 days each. The preparations were taken on 10 days together with a meal, on 10 days 3 hours after a meal. Blood samplings were performed 3 h after capsule ingestion for analysis of serum estradiol (S-E2), estrone (S-E1) and sex hormone binding globulin (SHBG). Before treatment, all women had climacteric complaints. During treatment these symptoms were alleviated and no discomfort was reported. No differences in serum levels of estrogens were found in either of the preparations when capsules were taken with or without food. However, serum levels of E2 were found to be 100% higher per mg substance given after E2COA vis-a-vis mE2. This indicates either a delayed breakdown and/or a better resorption. The E1/E2 ratio after E2COA was only half that after mE2 intake. This hints at another route of resorption. SHBG concentrations were somewhat elevated following mE2 administration, whereas a slight decrease was found after E2COA. The resulting post-treatment difference was significant, suggesting a less estrogenic liver effect by E2COA. No accumulation of E2 or E1 was seen after either of the preparations. Our findings support the hypothesis that E2COA, being fat soluble, is resorbed via the lymphatic system. By avoiding the first liver pass the dosage of estrogen can be halved. PMID- 2972163 TI - Dehydroepiandrosterone sulfate in saliva. AB - Concentrations of dehydroepiandrosterone sulfate (DHAS) in saliva and serum were determined in 11 healthy men aged 23-40 yrs, 55 healthy, non-medicated women aged 20-81 yrs and 13 healthy women aged 20-30 yrs, all taking combined oral contraceptives (OC). Serum DHAS was higher in men than in women of corresponding age and was negatively correlated to age in OC-free women. These sex- and age related differences were not found for salivary DHAS. Salivary and serum DHAS values were poorly correlated and the ratios between salivary and serum DHAS were variable, with a tendency to higher values in older subjects. Low saliva/serum DHAS ratios were found in OC users. The lack of a uniform, unequivocal relation between DHAS concentrations in saliva and serum renders salivary DHAS assays unsuitable for clinical purposes. PMID- 2972164 TI - Mothers of primary school children with Down's syndrome. How do they experience their situation? AB - The occurrence of anxiety or depression, experience of social support and feelings about the family situation were evaluated in 13 mothers of children of primary school age with Down's syndrome (DS). The results were compared with those obtained in a group of 13 females engaged in taking care of these children and assisting their families. Questionnaires were used to assess feelings of depression or anxiety (Hospital Anxiety and Depression Scale), to evaluate social support (Interview Schedule for Social Interaction) and the family situation (Family Adaptability and Cohesion Evaluation Scale). A semi-structured interview with the mothers was also conducted. The results indicated that negative feelings at the birth of a child with DS had almost invariably changed in a positive direction. Experience of depression or anxiety was uncommon. Social and emotional contacts were quantitatively normal, although more empathy was often desired. The families were relatively often described as enmeshed and controlled, but the experience of the family situation was generally positive. PMID- 2972165 TI - A double blind comparative study of remoxipride and thioridazine in the acute phase of schizophrenia. AB - This is the first comparative double blind study of remoxipride. Sixty-one patients with acute schizophrenia received either remoxipride (75-375 mg daily) or thioridazine (150-750 mg daily) for 6 weeks. There was no statistically significant between-drug difference in improvement in mental state, as measured by the Brief Psychiatric Rating Scale, although the trend favoured thioridazine; global assessment of illness severity at the last rating also favoured thioridazine. Sedation, anticholinergic effects, autonomic dysfunction and weight gain were significantly more common in patients receiving thioridazine. Both drugs produced few extrapyramidal effects, but both produced cardiovascular changes in two patients; neither drug produced significant abnormalities in laboratory tests. PMID- 2972166 TI - Abolishment of the positive feedback mechanism: a criterion for temporary medical hypophysectomy by LH-RH agonist. AB - The hypothalamic pituitary axis was studied in patients with an abnormal pattern of gonadotropin release during chronic treatment with LH-RH agonist. Two patients had PCOD and the third demonstrated the early luteinization phenomenon. Following a well-defined gonadotropin rise with initiation of LH-RH treatment, no further response was noted. Stabilization of the LH:FSH ratio in PCOD patients was noted after 4 weeks of treatment. Administration of both native LH-RH (100 micrograms) and intravenous pulsatile LH-RH did not evoke any rise in LH. In addition to the above LH-RH challenges, the positive feedback was examined by administration of estradiol benzoate (EB). The study demonstrated that, although the pituitary did not respond to any LH-RH challenge, it may still respond by a rise in LH following EB administration. Both functions of the hypothalamic pituitary axis should be examined in order to determine the state of medical hypophysectomy. PMID- 2972167 TI - Placebo-controlled comparison of danazol and high-dose medroxyprogesterone acetate in the treatment of endometriosis. AB - A prospective, double-blind, placebo-controlled study was designed to evaluate the clinical efficacy and tolerance of danazol and high-dose medroxyprogesterone acetate (MPA) in the treatment of mild-moderate endometriosis. After laparoscopical confirmation of endometriosis, 59 patients were randomized to receive danazol (200 mg 3 times daily), MPA (100 mg daily) or placebo for 6 months. Clinical examinations were done before and 1, 3, 6 and 12 months after the beginning of the study, and a 2nd laparoscopy 6 months after termination of the medication. Eighteen patients in the danazol group, 16 in the MPA group and 17 in the placebo group completed the trial. Total or partial resolution of peritoneal implants was observed in 60% of the patients receiving danazol and in 63% of the patients receiving MPA. In the placebo group, resolution was observed in 18%, while the size of the implants was estimated to be increased in 23% of the patients. In relation to placebo, danazol and MPA significantly alleviated endometriosis-associated pelvic pain, lower back pain and defecation pain, but they did not differ from each other in these actions. The appearance of acne, muscle cramps, edema, weight gain and spotting bleeding complicated MPA treatment. The present results indicate that because of good efficacy and tolerance, high-dose MPA is a useful alternative in the hormonal treatment of endometriosis. PMID- 2972168 TI - A relationship between sex hormone binding globulin and dehydroepiandrosterone sulfate in normally menstruating females. AB - Blood samples were collected from 120 normally menstruating women throughout the menstrual cycle. Serum was assayed for sex hormone binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS), estradiol (E2) and testosterone (T). There was an inverse relationship between SHBG and DHEAS, but no relationship between T or E2 and SHBG levels. There is at present no model which readily explains this inverse relationship between SHBG and DHEAS in normal subjects. PMID- 2972169 TI - Progesterone and progestins modulate beta-endorphin concentrations in the hypothalamus and in the pituitary of castrated female rats. AB - Hypothalamic and pituitary beta-endorphin (B-EP) concentrations are modified by ovariectomy and estrogen treatments, supporting a direct interaction between this peptidergic system and gonadal steroids. Because the use of progestins is becoming even more diffuse in clinical practice, we evaluated the effect of progesterone and of the synthetic progestins medroxyprogesterone acetate (MPA), norethisterone acetate (NET) and desogestrel on the concentration of B-EP in the medial-basal hypothalamus and the anterior and neurointermediate pituitary lobes in ovariectomized rats (OVX), treated or untreated with estradiol benzoate (EB). B-EP concentrations were significantly increased by desogestrel in the anterior lobe and by progesterone, desogestrel and medroxyprogesterone acetate in the neurointermediate lobe. Progesterone and progestins significantly reduced B-EP increase induced by estradiol benzoate in the anterior lobe. Estradiol benzoate treatment did not modify the effect of progesterone and desogestrel on B-EP in the neuro-intermediate pituitary lobe. Norethisterone acetate and progesterone increased B-EP concentrations in the medial-basal hypothalamus, while the other steroids were inactive. In contrast, in the hypothalamus all progestins attenuated the increase of B-EP induced by estradiol benzoate (p less than 0.01). These data indicate that progesterone and progestins modulate the hypothalamic and pituitary B-EP concentrations in concert with estrogens. The capacity of progestins to modify the hypothalamic contents of B-EP may represent one of the mechanisms of action of these steroids in influencing brain function. PMID- 2972170 TI - Bone turnover in postmenopausal women after withdrawal of estrogen/gestagen replacement therapy. AB - Bone turnover before and after withdrawal of estrogen/gestagen treatment was studied in a randomized trial with 110 healthy female volunteers, who had passed a natural menopause 6 months to 3 years before the start of the study. Urinary excretion of intravenously injected 99m-technetium diphosphonate was measured as an index of bone turnover; plasma bone Gla protein and serum alkaline phosphatase were measured as indices of bone formation; and fasting urinary excretion of hydroxyproline and calcium were measured as estimates of bone resorption. During 2 years of hormone treatment, all variables decreased highly significantly (p less than 0.001) to a constant low level. Three months after withdrawal all variables increased highly significantly (p less than 0.001) towards, but not above, pretreatment and placebo levels. We conclude that withdrawal of estrogen/gestagen replacement therapy in postmenopausal women increases bone turnover, but not in excess of pretreatment values. This indicates that bone loss (after withdrawal) is similar to that seen in the placebo group and that a rebound phenomenon is unlikely. PMID- 2972171 TI - Placebo-controlled comparison of danazol and high-dose medroxyprogesterone acetate in the treatment of endometriosis after conservative surgery. AB - To evaluate the clinical value of postoperative hormone therapy in endometriosis, 60 patients with advanced disease were randomized to receive in a double-blind study danazol (200 mg, 3 times daily), medroxyprogesterone acetate (MPA) (100 mg daily) or placebo post-operatively for 6 months. Treatment efficacy was evaluated clinically and at laparoscopy 6 months after medication. In relation to placebo, danazol and high-dose MPA treatments, which did not differ from each other in efficacy, significantly alleviated pelvic pain. In addition, the peritoneal endometriosis lesions found at 6-months laparoscopy were significantly smaller in the MPA and danazol groups than in the placebo group. Breakthrough bleeding, weight gain and acne complicated danazol treatment but only breakthrough bleeding complicated MPA treatment. These data suggest that postoperative treatment of advanced endometriosis with high-dose MPA or danazol is clinically beneficial. PMID- 2972172 TI - Pulsatile beta-endorphin release from the human pituitary in vitro. AB - An in vitro perifusion system was used to characterize spontaneous immunoreactive beta-endorphin (i beta-END) release from 10 human fetal (20-23 weeks gestation) and 2 human adult anterior pituitaries. Spontaneous i beta-END release from fetal anterior pituitaries was pulsatile, with a mean (+/- SE) pulse interval of 9.1 +/ 0.5 minutes, pulse amplitude of 120.8 +/- 46.1 pg with nadir to peak increment of 106.0 +/- 32.9%, and overall release rate of 209.7 +/- 65.0 pg/2 minutes. Blockade of calcium activity with 10 microM verapamil and 4 mM EGTA suppressed the frequency and amplitude of the spontaneous pulsatile i beta-endorphin release (n = 2). Administration of 2 nM human CRF for 20 minutes at the end of 2 perfusions induced 205 and 883% increases of i beta-END release over the preceding basal levels. Administration of 2 nM CRF for 50 minutes at the end of another perifusion led to a greater and prolonged increase (maximum 4620% relative to the immediately preceding basal level) in i beta-END release. Addition of 56 mM KCl during the last 20 minutes of this prolonged CRF stimulation further increased i beta-END release (to 7680% relative to the baseline preceding the CRF stimulation). Each of 4 quarters of adult anterior pituitaries (2 quarters each from 1 male and 1 female) also released i beta-END in a pulsatile fashion, with a pulse interval of 11.8 +/- 2.0 minutes, pulse amplitude of 7.4 +/- 0.8 ng with nadir to peak increment of 51.4 +/- 15.3%, and overall release rate of 21.7 +/- 2.9 ng/2 minutes. These studies demonstrate that i beta-END release from the isolated human anterior pituitary in vitro is characterized by high-frequency pulses, independent of hypothalamic stimulation. This spontaneous calcium-dependent pulsatile i beta-END release apparently reflects the activity of an intrapituitary pulse-generating mechanism. PMID- 2972173 TI - Rapid reduction of uterine leiomyomas with monthly injections of D-Trp6-GnRH. AB - Ten women with intramural leiomyomas were treated with the microencapsulated GnRH analogue Decapeptyl for 24 weeks. Four (4) mg Decapeptyl was injected, starting on day 21 of the menstrual cycle, and injections were repeated every 4 weeks for a total of 24 weeks. All patients showed a marked reduction in uterine size: before treatment it measured 284 +/- 57 cm3, after 8 weeks 122 +/- 33 cm3, and after 24 weeks 89 +/- 14 cm3. LH and estradiol decreased significantly; FSH decreased but not significantly; prolactin remained almost unaltered. Serum calcium, phosphate, alkaline phosphatase and osteocalcin increased, but, since calcium excretion (and hydroxyproline excretion) remained unaltered, these changes were considered to reflect increased bone turnover rather than bone loss. From these data it is concluded that Decapeptyl is very effective in reducing uterine fibroids, that treatment can be shorter than 6 months and that measurable bone loss did not occur. PMID- 2972174 TI - Transient expression of 5-HT1A receptor binding sites in some areas of the rat CNS during postnatal development. AB - The developmental evolution of 5-HT1A receptor binding sites was examined in the rat CNS during the early postnatal period using quantitative autoradiography and binding assays with 3H-8-OH-DPAT as the selective ligand. A progressive increase in the density of 5-HT1A sites was observed in the hippocampus, septum and cerebral cortex, up to adult levels which were reached around the third postnatal week. In contrast, complex biphasic (increase then decrease) changes were noted in other structures (for instance the nucleus of the lateral lemniscus), and even a progressive decrease in the density of 5-HT1A sites took place in the cerebellum during the first two postnatal weeks. The transient expression of 5 HT1A receptor binding sites in a structure such as the cerebellum which develops exclusively for the postnatal period further supports that 5-HT might play a trophic role during maturation of the CNS. PMID- 2972176 TI - Percutaneous transluminal renal angioplasty in the patient with a solitary functioning kidney. AB - Since the introduction of percutaneous transluminal renal angioplasty, it has become established as an effective treatment for the correction of renovascular hypertension or the preservation of renal function. However, there has been a general reluctance to use it or other interventional procedures in any patient with a solitary functioning kidney. We attempted renal angioplasty in 11 patients each of whom had a solitary functioning kidney. All 11 had had either a previous contralateral nephrectomy or an autonephrectomy from chronic arterial occlusion. The patients ranged in age from 42 to 81 years. In nine of the 11 patients, a catheter was negotiated across the stenosis, and the lesion was dilated successfully. In two, the lesion could not be crossed. In the group of nine patients whose lesions were dilated successfully, clinical follow-up (2-180 weeks) revealed that hypertension was cured in four and improved in five. Of the two patients in whom angioplasty was not possible, one required emergency renal bypass surgery and the other had no change in renal status. Renal function test results after the procedure were variable and did not correlate with decreases in blood pressure. We conclude that percutaneous renal angioplasty is a satisfactory alternative to surgery for the treatment of renovascular hypertension or the preservation of renal function in a patient with a solitary functioning kidney. PMID- 2972175 TI - Alpha interferon in T helper phenotype chronic lymphocytic leukemia: a report of three cases. AB - Three patients affected by T helper chronic lymphocytic leukemia were treated with low dose interferon alpha-2b (3 MU/m2 3 times weekly). The disease presented different pathologic expressions with diffuse skin lesions in one patient, a mild clinical course and a prolymphocytic variant with aggressive features, respectively, in the other two cases. A consistent response was observed within 3 6 weeks; by that time a reduction of blood and marrow lymphocytosis in the three patients and a regression of the cutaneous lesions were documented. Therefore, it should be emphasized that the use of alpha IFN, whose effectiveness on cutaneous T cell lymphomas has been already demonstrated, may represent an active agent in the treatment of leukemic T helper phenotype chronic lymphocytic proliferations. PMID- 2972177 TI - Second-trimester sonographic diagnosis of Down syndrome: role of femur-length shortening and nuchal-fold thickening. AB - Relative shortening of femur length and nuchal-fold thickening were evaluated as sonographic signs for the prenatal detection of Down syndrome. The sonograms of 28 fetuses identified by amniocentesis as having trisomy 21 were reviewed. The sonograms of 128 fetuses with a normal karyotype were evaluated as a control group. The finding of either an actual-to-expected femur-length ratio of 0.91 or less or a nuchal-fold measurement of 6 mm or more detected Down syndrome with a sensitivity of 42% and a specificity of 88%. The positive predictive value for Down syndrome detection when using femur-length shortening alone was greater than that when using the combination of femur-length shortening and nuchal-fold thickness. We conclude that femur-length shortening may be potentially useful for the detection of Down syndrome, whereas nuchal-fold thickening has a limited role. PMID- 2972178 TI - Relation of left ventricular shape to volume and mass in patients with minimally symptomatic chronic aortic regurgitation. AB - In 19 male patients with significant but minimally symptomatic or asymptomatic chronic aortic regurgitation, measurements of left ventricular shape, volume, and mass were assessed by cross-sectional echocardiography. End-diastolic volume and mass were significantly larger than normal values. There was a significant negative correlation between the volume-to-mass ratio (V/M) and both the end diastolic (r = -0.70, p less than 0.01) and end-systolic (r = -0.73, p less than 0.001) long axis-to-minor diameter ratios (L/D), suggesting that maintenance of the normal geometry of the left ventricle is in part dependent on the adequacy of hypertrophy, as expressed by a normal V/M. Since preoperative volume-to-mass indices have been shown to be important predictors of postoperative course, the L/D ratio should be a useful and simple parameter to follow in asymptomatic patients. PMID- 2972179 TI - Left atrial enlargement: an early sign of hypertensive heart disease. AB - Left atrial abnormality on the electrocardiogram (ECG) has been considered an early sign of hypertensive heart disease. In order to determine if echocardiographic left atrial enlargement is an early sign of hypertensive heart disease, we evaluated 10 normal and 14 hypertensive patients undergoing routine diagnostic cardiac catheterization for echocardiographic left atrial enlargement. All patients had normal coronary arteriography, sinus rhythm, normal left ventricular volumes and function, no valvular disease, and no echocardiographic or ECG left ventricular hypertrophy. No patient met ECG criteria for left atrial abnormality. The mean left atrial dimension was 3.46 +/- 0.3 cm in normal individuals versus 4.04 +/- 0.3 cm in the hypertensive patients (p less than 0.01). The left atrial index was also higher in the hypertensive group, 2.18 +/- 0.45 versus 1.88 +/- 0.10 cm/m2 (p less than 0.05), and the left atrial-to-aortic root dimension ratio was significantly higher in the hypertensive group, 1.36 +/- 0.20 versus 1.17 +/- 0.07 (p less than 0.01). We conclude that echocardiographic left atrial enlargement may be an early sign of hypertensive heart disease in patients with no other discernible cause of left atrial enlargement. PMID- 2972180 TI - Exaggerated secretion of atrial natriuretic polypeptide during dynamic exercise in patients with essential hypertension. AB - The plasma concentration of atrial natriuretic polypeptide (ANP) was measured in nine patients with essential hypertension during two grades of exercise tests performed in the supine position on a bicycle ergometer. The plasma ANP concentration significantly increased from 97.0 +/- 19.2 pg/ml to 107.6 +/- 23.7 pg/ml (p less than 0.05) during low-grade exercise (50% of the maximal heart rate) and from 96.2 +/- 16.5 pg to 192.8 +/- 30.7 pg/ml (p less than 0.01) during high-grade exercise (75% of the maximal heart rate). During high-grade exercise plasma epinephrine and norepinephrine concentrations showed significant increases. The plasma ANP concentration was significantly correlated with systolic blood pressure (r = 0.51; p less than 0.05). Patients with essential hypertension showed greater absolute increases in the plasma ANP concentration and systolic blood pressure during exercise compared to normotensive subjects. These results suggest that exercise stimulates secretion of ANP in response to its intensity in patients with essential hypertension and that a greater rise in atrial pressure, resulting from a greater elevation of systolic blood pressure, may be involved in the exaggerated secretion of ANP in patients with essential hypertension. PMID- 2972181 TI - Atrial natriuretic peptide release during atrial arrhythmias in cardiac transplantation. PMID- 2972182 TI - Relationship between ambulatory and exercise blood pressure and cardiac structure. AB - Recent research using automated ambulatory blood pressure recorders and echocardiography has revealed a closer relationship between LV structure and blood pressure during usual activity than with physician measurements of blood pressure. Because increased LV mass is a strong predictor of an adverse prognosis, this suggests that ambulatory blood pressure may also be more predictive of prognosis than casual readings, a view supported by preliminary evidence. Ambulatory blood pressure monitoring has already proved useful to identify patients with "white coat" hypertension or "office" hypertension, whose hearts appear normal by echocardiography. Further research is needed to determine the relative predictive value and comparative cost-effectiveness of ambulatory blood pressure monitoring and echocardiography for identification of patients in whom hypertension does or does not constitute a cardiovascular disease that requires antihypertensive therapy. PMID- 2972183 TI - A proposed occupational exposure limit for 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - One contaminant produced unintentionally during the manufacture of chlorophenols and phenoxy herbicides is 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The resulting TCDD-containing wastes have been detected at many hazardous waste sites which in recent years have been in the process of remediation. Concerns about worker exposure to TCDD-contaminated soil (dust) during remediation of hazardous waste sites have produced a need for an occupational exposure limit (OEL) for TCDD. The animal toxicology data and human experience with TCDD are reviewed, and an occupational exposure limit for TCDD is proposed. The animal data support risk estimations which are based on TCDD as a nongenotoxic carcinogen. Studies on human populations have failed to demonstrate clearly any significant long-term health effects at levels to which humans have been exposed. The data indicate that an 8-hr time-weighted average limit of 2 ng/m3 is appropriate, and the associated risk would be consistent with other carcinogens at their corresponding OELs. A preliminary OEL of 0.2 ng/m3 (200 pg/m3) is recommended, however, in light of other sources of exposure because of TCDD's ubiquitousness in the environment, its unclear mechanism of action, and its rather long biological half life in humans. This limit provides an ample margin of safety to prevent chloracne following repeated, acute exposure, and it addresses those chronic effects of TCDD observed in animal studies as well as those observed after accidental human exposure. The resulting body burden caused by chronic exposure to TCDD at the proposed OEL is examined. Its toxicological significance is compared with human tissue data and with other similarly persistent chemicals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972184 TI - Advances in cardiovascular technologies. AJC guide to equipment and supplies. PMID- 2972185 TI - Interventional cardiac catheterization at Duke Medical Center. PMID- 2972187 TI - Determinants of primary success in elective percutaneous transluminal coronary angioplasty for significant narrowing of a single major coronary artery. AB - Clinical and angiographic characteristics, procedural details and outcome were analyzed in 2,677 consecutive patients who underwent elective single-artery, single-lesion percutaneous transluminal coronary angioplasty (PTCA) between December 1980 and May 1987. Primary success was achieved in 2,479 (93%) patients. The primary success rate was significantly lower during the first period, when nonsteerable systems were used (73%), than in later periods (94%) (p less than 0.0001), when steerable and low-profile systems became available. Univariate analysis revealed the following variables as predictors of lower primary success: totally obstructed arteries (p less than 0.0001), presence of calcium in the narrowing (p = 0.002), prior myocardial infarction (p = 0.005), stenoses located in the right coronary artery (p = 0.02), narrowings between 90 and 99% in diameter (p = 0.02) and patients older than 60 years of age (p = 0.07). Multivariate analysis revealed the following 4 independent predictors of lower primary success: 100% obstruction (p less than 0.0001), calcium (p = 0.005), previous myocardial infarction (p = 0.029) and patients older than 60 years of age (p = 0.036). With present technology, single-narrowing elective PTCA can be performed with a high success rate in most patients. Although total occlusion, presence of calcium, older age and history of myocardial infarction influence the outcome unfavorably, PTCA can still be performed with acceptable primary success rates. PMID- 2972186 TI - Role of proto-oncogenes in myocardial hypertrophy. AB - A question of major clinical significance in cardiology is the nature of the signals that initiate and maintain the various types of myocardial hypertrophy, either in response to hemodynamic loading or in the absence of altered load. This review suggests that the proto-oncogene model, a concept derived from the study of cancer, can be very useful in identifying these signals. The proto-oncogene model conceives of cell growth regulation in terms of a limited number of classes of critical regulatory proteins: growth factors, growth factor receptors, intracellular transducing proteins and ribonucleic acid (RNA) transcription factors. Growth of all cells has dissociable components: hypertrophy (growth in size), deoxyribonucleic acid synthesis, mitosis and cytokinesis. Hypertrophy may be the end result of activation of RNA transcription. The various types of hypertrophy could reflect transcription of specific myocyte genes in response to different growth factors. At least 1 member of each functional class of proto oncogenes has been detected in the myocardium or myocytes, or both. The alpha 1 adrenergic receptor has been shown to be a growth factor receptor and to regulate RNA transcription. Continued work on proto-oncogenes in myocytes may open the way to manipulate the growth of these cells. PMID- 2972188 TI - Results of low speed rotational angioplasty for chronic peripheral occlusions. AB - After experimental investigation using postmortem human arteries, 19 patients with chronic peripheral artery occlusions were treated with a new angioplasty technique between December 1986 and October 1987. In 17 patients the superficial femoral artery and in 2 patients the popliteal artery were completely occluded. The length of the occlusions ranged between 5 and 25 cm (mean 11). The duration- estimated according to patients' history--was 5 to 48 months (mean 17). In 5 patients, durations of up to 30 months had been documented by angiography. A flexible, blunt, motor-driven rotating catheter was introduced through an 8 or 9Fr sheath and rotational angioplasty was performed at low speeds (up to 200 rpm). In 11 of 14 patients in whom this new technique was used as the primary intervention, the occlusions could be successfully reopened. In 2 patients after failure of conventional techniques the rotating catheter could not bypass the preexisting dissections in the same intervention. In 2 of 3 further patients after failure of conventional techniques the occlusions could be successfully reopened in a second intervention after several weeks. In none of the 19 patients did a perforation occur. It is concluded that with the new technique chronic peripheral artery occlusions can be reopened with a high success rate and without the danger of arterial wall perforation. The method can also be used in patients in whom conventional techniques have failed. PMID- 2972189 TI - Use of balloon flotation pacing catheters for prophylactic temporary pacing during diagnostic and therapeutic catheterization procedures. AB - The use of prophylactic temporary pacemakers during diagnostic catheterization, coronary angioplasty and percutaneous balloon valvuloplasty was investigated retrospectively over an 18-month period. Balloon flotation temporary pacemaker leads were placed in 193 (12%) of 1,609 patients undergoing diagnostic catheterization, 641 (65%) of 993 patients undergoing coronary angioplasty and 199 (100%) of 199 patients undergoing aortic or mitral valvuloplasty. There were no perforations or significant arrhythmic complications related to pacemaker placement in these 1,033 cases, and pacing was initiated promptly when required by withdrawal of the catheter tip into the right ventricle. Significant bradycardia or new conduction defects developed in 17 patients (1%) during diagnostic catheterization, 10 patients (1%) during angioplasty and 20 patients (10%) during valvuloplasty, but were severe enough to require initiation of temporary pacing in only 1 (0.06%), 4 (0.4%) and 5 (2.5%) patients, respectively. No patient undergoing diagnostic catheterization or angioplasty (but 5 patients undergoing valvuloplasty) required immediate pacing support to treat a life threatening bradycardia. The total cost of prophylactic pacemakers was $103,300, with a cost per actual use of $19,300 for diagnostic cases, $16,025 for angioplasty and $3,980 for balloon valvuloplasty. These data suggest that prophylactic temporary pacing is not indicated during either diagnostic catheterization or coronary angioplasty, but should be used routinely during balloon valvuloplasty. PMID- 2972191 TI - Accuracy of the RV6:RV5 voltage ratio for increased left ventricular mass. PMID- 2972190 TI - Early and long-term results of percutaneous transluminal coronary angioplasty in patients 70 years of age and older with angina pectoris. PMID- 2972192 TI - Phase II evaluation of menogaril in women with metastatic breast cancer after failure of first-line chemotherapy. AB - A total of 25 patients with metastatic breast cancer who had failed one prior chemotherapy regimen and had not received prior treatment with doxorubicin were treated with menogaril (200 mg/m2 i.v. over 1 h) every 4 weeks. Four patients (16%) achieved partial regressions lasting a median of 46 days. The median time to progression for all patients was 60 days and the median survival was 264 days. Seventeen patients subsequently received doxorubicin after removal from protocol and six (35%) achieved objective regression. We conclude that menogaril administered by the method that we employed has marginal activity in women with metastatic breast cancer after failure of prior chemotherapy. Failure to respond to menogaril does not preclude response to subsequent treatment with doxorubicin. PMID- 2972193 TI - Hairy cell leukemia and anti-leukocyte common antigen. AB - Thirty-three bone marrow biopsies from 15 patients with hairy cell leukemia (HCL) were evaluated morphologically and immunohistochemically by use of the peroxidase antiperoxidase technique to demonstrate reactivity for leukocyte common antigen (LCA). Hairy cells in all biopsies showed a distinctive and characteristic pattern for LCA, which decorated the periphery of the cytoplasm but that left most of the cytoplasm and the nucleus unstained. Anti-LCA was particularly helpful in highlighting focal or subtle leukemic infiltrates. Hairy cells in biopsies from three patients had, on routine morphologic examination, a spindled and sarcomatoid appearance, but these too were strongly LCA positive. Treatment regimens were variable: five patients had splenectomy and received chemotherapy; five patients had splenectomy alone; and four patients had chemotherapy alone. Seven patients received interferon, and one patient received no treatment. In those six patients who had multiple biopsies as part of follow-up examinations, hairy cells as identified by anti-LCA were continuously present. Often in significant numbers, and were usually underestimated or not identified by routine examination. In those patients who received chemotherapy, qualitative alterations in the LCA reaction of hairy cells were observed. PMID- 2972194 TI - Pseudo-Gaucher cells preceding the appearance of immunoblastic lymphoma. AB - A 49-year-old white woman who presented with multiple lytic bone lesions was found to have Gaucher-like storage cells in multiple bone marrow aspirates and a percutaneous bone marrow biopsy several months before the development of overt disseminated lymphoma. Open bone biopsies of three different sites at initial presentation revealed only necrotic debris and a nondiagnostic polymorphic infiltrate. Elevated leukocyte beta-glucocerebrosidase levels ruled out the diagnosis of classic Gaucher's disease. The ultrastructural characteristics of these storage cells were found to be identical to those of pseudo-Gaucher cells found in patients with chronic myelogenous leukemia and distinctly different from those previously reported in other non-Hodgkin's lymphoma or plasma cell dyscrasias. A possible pathogenetic mechanism is suggested. PMID- 2972195 TI - Radioimmunoassay of beta-thromboglobulin. False high values from frozen plasma samples. AB - Aliquots of plasma samples assayed after storage at -26 degrees C can show a significantly higher level of measured beta-thromboglobulin (beta-TG) than the corresponding unstored aliquots. Blood spun at the recommended speed of 1,800 X g still contains platelets that will release beta-TG on freezing to yield false results--a point that has not been stressed in the literature. Aliquots of blood plasma spun at either 1,800 X g (low speed [LS]) or 20,000 X g (high speed [HS]) were assayed by radioimmunoassay fresh and after storage at 4 degrees C or -26 degrees C. Storage at -26 degrees C increased mean beta-TG values of fresh or 4 degrees C stored LS samples from 0.78 to 1.94 nmol/L (28-70 ng/mL), whereas all HS mean values were +/- 0.51 nmol/L (between 18 and 19 ng/mL). The authors believe that in addition to the accepted precautions regarding blood sampling and handling, it is essential to stress that for accurate measurement of beta-TG in plasma, samples should be centrifuged at HS or, if this is not feasible, that they be assayed as soon as possible without prior freezing. PMID- 2972196 TI - Deep venous thrombosis, inflammatory bowel disease, and protein S deficiency. AB - The authors observed a patient with severe inflammatory bowel disease (IBD) complicated by ten episodes of deep venous thrombosis (DVT). Her unusual thrombotic tendency prompted further investigation for primary hypercoagulability. A nonfamilial deficiency of Protein S was documented suggesting the deficiency was acquired. This occurrence suggests that low Protein S levels should be considered as a potential etiologic factor in patients with IBD and recurrent DVT. PMID- 2972198 TI - Treatment of renovascular hypertension with percutaneous transluminal renal angioplasty. PMID- 2972197 TI - Neonatal and maternal platelets: activation at time of birth. AB - Determination of the plasma concentrations of beta-thromboglobulin (BTG), thromboxane B2 (TxB2) and platelet factor 4 (PF4) were made at the time of birth in 18 newborns and their respective mothers. Both groups show significant elevation of all these molecular markers, suggesting marked platelet activation. The elevated TxB2 levels in the newborn group, 25 +/- 8 pg/ml, are compatible with a normally functioning and activated platelet prostaglandin pathway. Mode of delivery, vaginal or caesarean section, does not significantly influence the degree of activation in either group. Ultrastructural platelet examination did not reveal any morphologic differences between maternal and newborn platelets. There appears to be marked activation of the newborn and maternal platelet systems at the time of birth, and we postulate that this may explain in part the transient platelet dysfunction observed in newborns. PMID- 2972199 TI - Effect of increasing intraabdominal pressure and volume on left ventricular function in continuous ambulatory peritoneal dialysis (CAPD). AB - To determine the effect of increasing intraabdominal pressure on left ventricular (LV) systolic function in patients undergoing continuous ambulatory peritoneal dialysis (CAPD), we studied 15 patients who had been on CAPD for at least 2 months. The study protocol included measurement of echocardiographic LV internal dimensions in diastole and systole, LV wall thickness and mass, LV ejection time, the mean velocity of circumferential fiber shortening (mean Vcf), heart rate, BP, and intraabdominal pressure. Measurements were obtained at baseline (OL) and following intraperitoneal infusion of 1-L increments of dialysate solution (up to 4 L) in both the recumbent and standing positions. Significant differences in LV systolic function resulting from intraperitoneal infusion of dialysate were confined to the subgroup with increased LV wall thickness (n = 8). In this subgroup, mean Vcf decreased progressively from 0.99 +/- 0.18 circumferences/second at baseline to 0.88 +/- 0.16 circumferences/second at 1 L (P = NS), to 0.86 +/- 0.16 circumferences/second at 2 L (P = NS), to 0.66 +/- 0.18 circumferences/second at 3 L (P less than 0.005), and to 0.60 +/- 0.14 circumferences/second at 4 L (P less than 0.005) in recumbent position. Measurements obtained in the standing position paralleled those observed during recumbency. These changes were accompanied by a significant decrease in the mean LV internal dimension in diastole and a significant increase in intraabdominal pressure, but no significant change in the mean LV internal dimensions in systole, mean heart rate, or mean systolic or diastolic BP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972200 TI - Application of DNA-DNA hybridization of dual labeled probes to the detection of trisomy 21, monosomy 21, and sex determination. AB - Chromosomal aneuploidy is usually identified by cytogenetic methods. However, for some purposes it would be desirable to have an easier method of recognizing specific trisomies or monosomies. We have devised such an assay. It involves the simultaneous hybridization of two chromosome-specific DNA probes labeled with different isotopes (32P and 35S) to human chromosomal DNA immobilized on a nitrocellulose filter. The hybridization signals are then counted, and the dpm ratios between the two isotopes are determined. In this paper we show the application of this technique in sex determination, in the analysis of two human cell lines reported to have chromosome 21 monosomy, and, especially, in testing for Down syndrome. PMID- 2972201 TI - Maternal smoking and Down syndrome. PMID- 2972202 TI - Hyponatremia and atrial natriuretic peptide secretion in patients with vasopressin-induced antidiuresis. PMID- 2972203 TI - Aneuploidy and the fragile X syndrome. AB - The possibility that female carriers of the fragile X gene(s) are at increased risk for nondisjunctional events leading to aneuploid offspring has been suggested by several investigators. To better address this question we analyzed pedigrees of 117 families in which the fragile X syndrome is segregating. The 117 pedigrees, originally collected for segregation analyses, included 236 females with offspring whose carrier status was determined by cytogenetic or pedigree analysis or by analyses using flanking DNA markers. These 236 females have had 931 offspring including one 47,XXY and 6 trisomy 21 individuals (1/155). Statistical analysis suggested that the observed rate of trisomy 21 was significantly higher than expected (Fisher's exact test, p less than or equal to 0.05). Assuming a Poisson distribution to calculate the confidence interval for the observed rate of trisomy 21 individuals, we found that the expected rate of 1.6/1000 in this sample fell outside the 99% confidence limits of our observed rate of 1/155. Additional data from a larger sample are needed to replicate these findings. PMID- 2972204 TI - Crown size asymmetry in males with fra (X) or Martin-Bell syndrome. AB - We measured the mesio-distal and bucco-lingual crown diameters of 13 males with the fra(X) or Martin-Bell syndrome. Fluctuating crown-size asymmetry was calculated and compared with values obtained in normal Caucasian children and also with a sample of 19 males with Down syndrome. A statistically significant increased asymmetry was found in the fra(X) males when compared to normal control individuals. In the maxilla, Down syndrome males showed a significantly higher tooth crown asymmetry than fra(X) males. (less than 0.02); in the mandible, no significant differences were found between the 2 groups. It is suggested that crown size asymmetry be included in the evaluation of fra(X) males. PMID- 2972205 TI - Recent experience in prenatal fra(X) detection. AB - At least 35 cases of prenatal fra(X) diagnosis have been confirmed and reported. Amniotic fluid, fetal blood and chorion -ic villus samples have exhibited fra(X) (q27.3) in cultures from 26 males and 9 females. Here we have detected fra(X) in female and male amniotic fluid specimens, AF1/fra(X),X and AF2/fra(X),Y, respectively, and a male CVS/fra(X),Y using both FUdR and excess thymidine (THY) to demonstrate the marker chromosome. Both FUdR and THY detected fra(X) and usually FUdR was superior to THY with the exception of placental cultures. It was important to examine more than one culture per protocol since no fra(X) was observed in one AF2 FUdR culture while another exhibited 19.2% expression. Similarly, confirmation studies in lung fibroblast cultures for AF2 exhibited 4.3% fra(X) in one lab while another found negative results. A similar observation in whole blood cultures was also made recently by us. In addition, we have recently experienced our first false negative fra(X),X prenatal diagnosis. We have observed another case where only one cell in 300 exhibited fra(X) where the male fetus was 50% at-risk and was referred to us after the 20th week of gestation by sonography. On the basis of our experience we recommend the following: 1) the excess THY fra(X) induction system is effective but not superior to FUdR; 2) at least two duplicate cultures per induction system should be analyzed for the marker chromosome to avoid the possibility of false-negative diagnosis; 3) where fra(X) is not demonstrated or is present in very low frequencies in CVS and/or amniotic fluid cultures, complementary DNA marker studies and/or fetal blood cultures must be made available; 4) gestational age dating by ultrasonography is recommended as early as possible. PMID- 2972206 TI - Fragile X expression in short-term whole blood cultures is affected by cell density. AB - The effect of cell density on expression of the fragile site at (X)(q27.3) in short-term whole-blood cultures from patients with fragile X [fra(X)] or Martin Bell syndrome was studied. A significant increase in fra(X) frequency was observed in 7 of 8 samples when cell density was decreased. Higher fra(X) frequency was not always noted at below-routine density, but in some cases fra(X) expression was depressed at above-routine density. We conclude that decay of the FUdR effect explains the fact that fra(X) expression is affected by culture density. It is significant that a relationship exists between the two; it suggests that in order to maximize fra(X) expression in cases with low-percentage fra(X) with standard methods, cell density may have to be adjusted. It is possible that in individuals who are normally nonexpressing, such as some obligate female carriers and nonpenetrant males, fra(X) expression may be sensitive to cell density effects. PMID- 2972208 TI - Down syndrome and femur length. PMID- 2972207 TI - Aortic hypoplasia and cardiac valvular abnormalities in a boy with fragile X syndrome. AB - An 18-year-old mentally retarded male with the Martin-Bell syndrome was fragile X positive. He died suddenly with viral pneumonia and myocarditis. At autopsy, generalized tubular hypoplasia of the aorta and a mild coarctation were discovered. The base of the mitral and tricuspid valves showed striking aberrations in elastin distribution and structure by light microscopy. Local collagen alterations were also noted. Comparable changes were seen in the skin elastin as well as a severe depletion of acid mucopolysaccharides. These changes suggest a structural disruption underlying the clinical connective tissue problems in some patients with the fragile X syndrome. PMID- 2972210 TI - Comparison of different modalities of treatment for minimal and mild endometriosis. AB - This is a retrospective study designed to compare the results of treatment of minimal and mild types of endometriosis with danazol alone, operative laparoscopy and danazol combined, and operative laparoscopy alone. The population consisted of 112 patients with minimal endometriosis and 126 patients with the mild type. Pregnancy rate and endometriosis resolution were lowest in patients who were treated with danazol alone and highest in patients treated with operative laparoscopy alone. It is concluded that in an infertile population with minimal to mild endometriosis danazol alone appears ineffective in improving the pregnancy rate or in achieving complete resolution of endometriosis. Laparoscopic surgery for the treatment of minimal to mild endometriosis seems to be a promising method of treatment. PMID- 2972209 TI - Effects of smoking on serum lipoproteins and bone mineral content during postmenopausal hormone replacement therapy. AB - We examined the effect of smoking on the treatment response in serum estrogens, serum lipids and lipoproteins, and bone mineral content in 110 postmenopausal women treated for 2 years with either percutaneous or orally administered combined hormones or placebo and followed up by examinations every 3 months. Serum estradiol and estrone levels during oral hormone administration were lower in smokers than in nonsmokers, whereas no differences related to smoking habits were observed during percutaneous hormone administration. Serum total and low density lipoprotein cholesterol were significantly reduced in both smokers and nonsmokers receiving hormones, but the response in smokers was only half that observed in nonsmokers (not significant). When the impact of the route of hormone administration was examined in relation to smoking habits, the response in serum total and low-density lipoprotein cholesterol in smokers receiving oral hormones was significantly lower (p less than 0.05) than that observed in nonsmokers. No differences in serum lipids or lipoproteins were observed between smokers and nonsmokers receiving percutaneous hormones. The response in bone mineral content in smokers and nonsmokers receiving percutaneous hormones and placebo was not significantly different, although the overall response differed significantly in the two groups (p less than 0.001). In contrast, the response in smokers receiving oral hormones was significantly lower than that observed in the corresponding nonsmokers (p less than 0.01). We conclude that smoking greatly affects the response on circulating levels of estrogens in postmenopausal women treated with orally administered hormone replacement therapy and that the subsequent treatment response on serum lipids and lipoproteins and on bone mineral content is reduced accordingly. The study suggests that alternative routes of administration should be considered when postmenopausal estrogen therapy is instituted in women who smoke. PMID- 2972212 TI - Effect of vasopressin on atrial natriuretic peptide release and renal function in dogs. AB - To assess the effect of arginine vasopressin (AVP) and 1-deamino-8-D-AVP (DDAVP) on atrial natriuretic peptide (ANP) release, renal water and electrolyte excretion, and cardiovascular function, AVP and DDAVP were administered at a dose of 10 ng.kg-1.min-1 iv for 30 min into anesthetized dogs receiving saline infusion at a rate of 1 ml.kg-1.min-1 (n = 12). In the control study, saline was infused alone (n = 6). AVP potentiated the plasma ANP response to an increase in plasma volume produced by saline infusion, increased mean arterial blood pressure (MAP), and exaggerated the natriuresis and kaliuresis. DDAVP did not potentiate the increase in plasma ANP but enhanced the natriuresis without any rise in MAP. Saline alone increased plasma ANP as well as sodium and potassium excretion with no changes in MAP. Inulin and p-aminohippuric acid clearances did not change during these studies. The results suggest that in hydrated dogs, AVP may increase ANP release and arterial blood pressure via the vasopressor activity of AVP and potentiate the natriuresis and kaliuresis, but the increased ANP may play little role in the natriuresis. PMID- 2972211 TI - Effects of hypercalcemia and ouabain on plasma atrial natriuretic polypeptide in anesthetized dogs. AB - For the purpose of examining the role of calcium ion in the secretory process of atrial natriuretic polypeptide (ANP), we studied the effects of hypercalcemia and ouabain on plasma concentration of immunoreactive ANP (irANP). Pentobarbital sodium-anesthetized dogs were treated with calcium chloride infusion (0.136 mmol.kg-1.min-1, 10 min) or ouabain injection (30 micrograms/kg), and plasma irANP concentration, right atrial pressure, mean arterial pressure, heart rate, and serum calcium concentration were measured. To evaluate the effect of rising arterial pressure, plasma irANP concentration was also measured in dogs treated with phenylephrine hydrochloride (10 micrograms/kg). With calcium chloride infusion, serum calcium concentration and plasma irANP concentration, respectively, increased to about three times and about four times their basal levels. This increase of plasma irANP concentration was not attenuated by pretreatment with adrenoceptor blockers. Ouabain increased plasma irANP concentration to approximately 2.5 times the initial level. Neither calcium chloride nor ouabain produced any effect on right atrial pressure and heart rate, but both significantly increased mean atrial pressure. Phenylephrine caused a greater increase in mean arterial pressure than both calcium chloride and ouabain. However, there was no significant increase in plasma irANP concentration. These results suggest that the calcium ion may play a key role in the secretory process of ANP. PMID- 2972213 TI - Distinct receptors for insulin-like growth factor I in rat renal glomeruli and tubules. AB - Purified preparations of renal glomeruli and tubules were obtained by a procedure involving perfusion of rat kidneys with magnetic iron oxide particles to selectively separate the iron-containing glomeruli from the nonmagnetic tubules. Detergent-soluble extracts of both renal glomerular and tubular membranes showed high-affinity, specific binding of 125I-labeled insulin-like growth factor I (125I-IGF-I), whereas degradation of this peptide hormone was minimal during a 90 min incubation at 22 degrees C in the presence of 2.5 mM EDTA and 5 mM N ethylmaleimide. The affinity of these receptors for IGF-I appeared identical in the two types of renal tissue, since 50% inhibition of 125I-IGF-I binding to both glomerular and tubular tissue occurred in the presence of approximately 3 x 10( 9) M unlabeled IGF-I. In contrast, insulin was much less effective at blocking 125I-IGF-I binding to either tissue, with 1 x 10(-6) M insulin required to produce 50% inhibition of binding. Relative to 125I-IGF-I binding, 125I-insulin binding to glomerular and tubular tissue was significantly lower per milligram protein. 125I-IGF-I was specifically cross-linked to a glomerular receptor subunit that migrated as two discrete bands with relative molecular weight (Mr) of 140,000-150,000 on sodium dodecyl sulfate polyacrylamide gels in the presence of 40 mM dithiothreitol. In contrast, 125I-IGF-I was cross-linked to a tubular receptor subunit that migrated as two discrete bands but at a slightly different position, with Mr of 120,000-140,000.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972214 TI - Effect of puromycin aminonucleoside on HSPG core protein content of glomerular epithelial cells. AB - It has been suggested that the glomerular basement membrane heparan sulfate proteoglycan (HSPG) is an important determinant of the glomerular permselectivity barrier. Derangements in the content of basement membrane heparan sulfate have been implicated in alterations in glomerular permselectivity seen in many glomerular diseases such as aminonucleoside nephrosis. The cellular origin and metabolism of the glomerular basement membrane HSPG have not been studied in detail. We have detected the expression of the proteoglycan by cloned glomerular visceral epithelial cells of the rat by employing a specific antibody against the core protein of HSPG isolated from the rat glomerular basement membrane. These findings suggest that in the rat in vivo glomerular visceral epithelial cells are one source of heparan sulfate present in the glomerular basement membrane. The effect of puromycin aminonucleoside (PAN) on the HSPG core protein content of the cloned glomerular epithelial cells was studied. By a quantitative immunoperoxidase method, the aminonucleoside caused a 28% reduction in the core protein content of the epithelial cells (P less than 0.01) following 72 h of incubation. However, the content of Heymann nephritis-related antigen, Fx1A was unchanged. Studies employing [3H]leucine incorporation showed that PAN was a weak inhibitor of de novo protein synthesis at 24 h of incubation, with complete recovery at 48 and 72 h. These data suggest that PAN effect on heparan sulfate core protein cannot be attributed to generalized inhibition of protein synthesis. The precise mechanism underlying the aminonucleoside effect on heparan sulfate core protein remains to be elucidated. PMID- 2972215 TI - ANF inhibits vasopressin-induced Ca2+ mobilization and contraction in glomerular mesangial cells. AB - It has been suggested that atrial natriuretic factor (ANF) may increase glomerular filtration rate (GFR) via alteration of the glomerular ultrafiltration area, which is at least in part regulated by the contractile state of mesangial cells. The present study therefore investigated the effect of ANF (atriopeptin III) on arginine vasopressin (AVP)-induced Ca2+ mobilization and contraction in cultured rat mesangial cells. AVP (10(-8) M) increased intracellular Ca2+ ([ Ca2+]i) as measured by the quin2 method (142.1 +/- 3.7 vs. 297.9 +/- 8.9 nM, P less than 0.001). This effect, however, was diminished by 10(-8) M ANF (297.9 +/- 8.9 vs. 233.1 +/- 9.3 nM, P less than 0.0025). ANF alone did not affect basal [Ca2+]i. AVP-stimulated 45Ca2+ efflux (cpm.mg prot-1.30 s-1) from mesangial cells within 30 s (1,432 +/- 70 basal vs. 2,751 +/- 287 AVP at 30 s, P less than 0.001). This effect of AVP on 45Ca2+ efflux was also inhibited by ANF (1,403 +/- 62 basal vs. 1,584 +/- 87 AVP and ANF, NS); ANF alone had no effect on spontaneous 45Ca2+ efflux. Furthermore, ANF inhibited AVP-induced 45Ca2+ uptake, which is purportedly one of the major determinants of the sustained phase of cell contraction. Cell contraction was assessed by digital imaging analysis. AVP (10( 7) M) caused contraction of 33.8 +/- 2.0% of the cells, but the contractile response decreased to 20.7 +/- 2.3% in the presence of 10(-9) M ANF and was virtually abolished in the presence of 10(-8) M ANF. Guanosine 3',5'-cyclic monophosphate (cGMP) has been proposed as the second messenger for ANF. ANF (10( 7) M) stimulated cGMP production significantly (0.54 +/- 0.12 vs. 22.30 +/- 5.91 pmol.mg prot-1.10 min-1, P less than 0.01), but it had no effect on cGMP at concentrations of 10(-8) M and below. 8-Bromo-cGMP (8-BrcGMP) attenuated the stimulatory effect of AVP on [Ca2+]i, 45Ca2+ efflux, and 45Ca2+ uptake. AVP induced cell contraction was also decreased in the presence of 2 mM 8-BrcGMP (33.8 +/- 2.0% vs. 18.8 +/- 2.6%, P less than 0.005). The present results suggest that ANF may increase GFR in part by blocking the cellular action of AVP and possibly other vasoconstrictors on glomerular mesangial cells, thus affecting glomerular filtration area. A role of cGMP as second messenger for ANF is possible, but it appears unlikely to be the only mediator for the effects of ANF. PMID- 2972217 TI - Release of atrial natriuretic factor in hypophysectomized rats. AB - The purpose of these studies was to examine the effect of hypophysectomy (HYPOX) on the secretion of atrial natriuretic factor (ANF) and related changes in renal function. Pentobarbital sodium-anesthetized rats received a 20% intravenous blood volume expansion. In intact rats (n = 7), plasma ANF was 39 +/- 3 pg/ml before volume expansion and increased to 71 +/- 10 and 77 +/- 7 pg/ml after volume expansion (P less than 0.01). Volume expansion in HYPOX rats (n = 8) also resulted in an increase in plasma ANF concentration that was significantly less than in the intact group. With volume expansion, sodium excretion rate increased from 0.19 +/- 0.06 to 1.43 +/- 0.43 microM. min-1.100 g body wt-1 (P less than 0.01) in intact rats and from 0.24 +/- 0.08 to 0.74 +/- 0.14 microM.min-1.100 g body wt-1 (P less than 0.05) in HYPOX rats (HYPOX less than intact; P less than 0.05). In an isolated heart-lung preparation, HYPOX rats secreted significantly less ANF than intact rats during two 30-min perfusion periods (P less than 0.02). Atrial ANF concentration was also significantly less in HYPOX rats (P less than 0.05). These results show that hypophysectomy leads to an attenuation of the ANF response to atrial distention and attenuated natriuretic response to blood volume expansion and suggest that the pituitary is required to maintain normal cardiac endocrine function. PMID- 2972216 TI - Vascular vasopressin receptors in renal hypertensive rats. AB - We investigated the plasma concentration of [Arg8]-vasopressin (AVP), the density of AVP-binding sites on membranes from the mesenteric vascular bed, and the pressor response to AVP of the perfused mesenteric vasculature in vitro from one kidney, one-clip (1K, 1C) and two-kidney, one-clip (2K, 1C) Goldblatt hypertensive rats. The plasma concentration of AVP was increased in 1K, 1C hypertensive rats. The density of AVP-binding sites was similar in sham-operated normotensive, in 2K, 1C hypertensive, and in uninephrectomized rats but was significantly decreased in 1K, 1C hypertensive rats (P less than 0.05). The binding affinity of AVP was similar in all experimental groups. Vasoconstrictor response to AVP was increased in 2K, 1C hypertensive rats (27% higher than sham operated normotensive rats, P less than 0.05). Responses in 1K, 1C hypertensive rats were similar to those of uninephrectomized rats. Our results indicate that together with an increased concentration of AVP in plasma the number of vascular AVP-binding sites is decreased in 1K, 1C hypertensive rats, whereas both are unaltered in 2K, 1C hypertensive rats. Vascular AVP receptors appear to be regulated inversely to plasma AVP concentrations. Pressor responsiveness to AVP is normal in 1K, 1C hypertensive rats and exaggerated in 2K, 1C hypertensive rats. Increased vascular responsiveness to AVP may occur independently of the regulation of AVP receptors and may contribute to elevation of blood pressure in renal hypertension in the rat. PMID- 2972218 TI - Pathophysiology of experimental leishmaniasis: the role of parasite physiology in the development of metastatic disease. AB - This paper addresses the issue of how physiological properties of Leishmania determine the pattern of development of disseminated leishmaniasis in the mammalian host. It presents direct experimental evidence from in vivo studies that species of Leishmania differ in their capacity to multiply in cutaneous and visceral sites which results in differences in the pattern and rate of development of leishmaniasis. It was found that Leishmania mexicana amazonensis begins to multiply in the cutaneous site of inoculation within 7 days. Parasites, detected in the liver and spleen at 4 weeks, increased 100-fold during the next 4 months. However, the slow multiplication of L. mexicana amazonensis in the liver and spleen was more apparent than real. Parasites implanted in those organs of athymic nude mice by an intravenous injection were rapidly eliminated with a half time of 16 hr. Thus, the parasites found in small numbers in the liver during the development of disseminated cutaneous disease in mice are most likely those which have been recently removed from the blood. Those few parasites that are not removed from the blood can establish metastatic foci in distant cutaneous sites, and replicate progressively once there. PMID- 2972219 TI - [Developmental defects of the uterus and their surgical correction to restore the child-bearing function]. PMID- 2972220 TI - [The importance of laparoscopy in the diagnosis of chronic salpingo-oophoritis]. PMID- 2972221 TI - [Prevention of hemodynamic disorders during diagnostic and surgical laparoscopy in women suffering from sterility]. PMID- 2972222 TI - Mononuclear cell patterns in the skin. An immunohistological and morphometrical analysis. AB - A large variety of skin diseases is characterized by the presence of mononuclear cell infiltrates in the dermis and to some extent in the epidermis. We have investigated frozen material of 566 lesions of benign and malignant skin diseases by immunohistological methods with a panel of 20 monoclonal antibodies; quantitative studies using computer-assisted image analysis were additionally performed in 80 specimens. In all reactive lesions, mononuclear cells were arranged in distinct compartments simulating the architecture of normal lymphatic tissue. A qualitatively uniform T cell compartment (mainly helper-inducer T lymphocytes, suppressor-cytotoxic T lymphocytes, Langerhans' cells) with slight quantitative differences was found in all inflammatory skin diseases, in peritumoral infiltrates, and also in normal skin. The B cell compartment (B lymphocytes, few T lymphocytes and monocytes) is only rarely seen (some B cell lymphomas and pseudolymphomas). The monocyte compartment (monocytes, few T lymphocytes) associated with a T cell compartment is typical for granulomatous skin lesions. The epidermis forms a separate functional region, which might evolve into a lymphoepithelial compartment in diseased skin. Low-grade malignant lymphomas of the skin display similar architectural patterns as reactive lesions, whereas high-grade malignant lymphomas do not. Obviously the mononuclear cells in the skin are usually arranged in compartments and form a limited number of distinct patterns. As similar patterns are found in diseases that are completely different with respect to clinical appearance, histological features, cause, and outcome, the patterns are not disease specific but reflect general anatomical functional relationships of inflammatory cells and the skin. PMID- 2972223 TI - A comparison of the response of hand and facial muscles to non-depolarising relaxants. AB - This study compares the response of the muscles of the hand to those of the face during non-depolarising neuromuscular blockade. It was found that observation of the facial muscles was misleading both in terms of degree of neuromuscular blockade and adequacy of reversal. PMID- 2972224 TI - Sufentanil to supplement nitrous oxide in oxygen during balanced anaesthesia. AB - Sufentanil in doses of 0.4 and 1.0 microgram/kg was used as an analgesic supplement to nitrous oxide-oxygen anaesthesia in 28 patients whose lungs were ventilated during body surface or lower abdominal surgery. Significant decreases in arterial blood pressure followed drug administration (mean 35.6%). Decreases in arterial pressure of greater than 30% occurred in 5 out of 14 patients in the low dose group, and 12 out of 14 in the high dose sufentanil group. There was a blunting of the pressor response to laryngoscopy and intubation after sufentanil. Increases in blood pressure were seen in both treatment groups at the onset of surgery. The peak responses were 27% increase in mean arterial pressure in the low dose group, and 51% increase in the 1.0 microgram/kg group. There was however no significant change in heart rate. Fourteen patients (8 in the low dose group) required volatile supplementation during surgery for clinical signs of inadequate anaesthesia. This occurred between 21 and 70 minutes after sufentanil administration, and plasma drug concentrations 0.06 and 0.54 ng/ml. Tracheal extubation was successfully achieved at the end of surgery in 26 out of 28 patients at plasma sufentanil concentrations which ranged from 0.03 to 0.37 ng/ml. The two remaining patients, neither of whom had received additional volatile supplementation, required naloxone to overcome postoperative ventilatory depression. No cases of intra-operative awareness were noted. PMID- 2972225 TI - The Downs' adjustable flow generator. AB - A venturi powered device for the generation of continuous positive airways pressure has been assessed. Some of its functional characteristics appear to be ideal, but the oxygen concentration and adequacy of flows require checking after each adjustment. A positive pressure relief valve and an inlet valve are necessary for the safe use of the system and an airway pressure monitor is recommended. PMID- 2972226 TI - Postoperative pain: a comparison of laparoscopic sterilisation and diagnostic laparoscopy. PMID- 2972227 TI - Flushing between thiopentone and atracurium. PMID- 2972228 TI - Pruritus with sublingual buprenorphine. PMID- 2972229 TI - Developmental transitions of myosin isoforms and organisation of the lateral muscle in the teleost Dicentrarchus labrax (L.). AB - In Dicentrarchus labrax (the sea bass) the differentiation of lateral muscle fibres occurs at different stages and in different ways in the superficial (red), intermediate (pink) and deep (white) regions of the myotome. At hatching the myotomes are composed of presumptive white and red fibres, the latter forming a superficial monolayer present only near the transverse septum. At this stage, differences between the fibre types are mainly ultrastructural. From their different reactions with isoform-specific antibodies to mullet myosin, and the appearance of histochemical mATPase activity, it appears that in both red and white muscle fibres there is a transition in myosin composition from an early larval form (L1R and L1W respectively) to a late larval form (L2R and L2W) and then to the isoforms typical of adult red and white muscle. The transition from L1W to L2W in the deep muscle occurs very rapidly and early in larval life (between 10 and 28 days), whereas the equivalent transition in the superficial muscle (from L1R to L2R) is a gradual process beginning in fibres near the transverse septum and spreading hypo- and epi-axially as this layer grows around the deep muscle. The definitive adult forms (AR and AW), distinguishable by the appearance of characteristic histochemical myosin ATPase activity, are present in the superficial red muscle by 80 days, but later in the deep white muscle (by 20 months), respectively. Compared to the superficial red and deep white muscle, the intermediate (pink) muscle layer first appears relatively late (80 days), but then acquires the histo- and immunohistochemical profile characteristic of the adult form much more rapidly.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972230 TI - Development of immunoreactive atrial natriuretic peptide in fetal hearts of spontaneously hypertensive and Wistar-Kyoto rats. AB - The development of immunoreactive atrial natriuretic peptide (ANP) was studied in fetal hearts of spontaneously hypertensive (SHR) and compared to normotensive Wistar-Kyoto (WKY) rats. While SHR fetal hearts were noticeably less developed than those of WKY at 10 and 11 days gestation, both strains showed ANP immunoreactive cells in some but not all primitive heart tubes. At 12 days additional ANP immunoreactive cells appeared in formative trabeculae of the ventricle and atrium. ANP cells were also observed in the myogenic layer of the truncus and bulbus arteriosus and their derivatives from 11 through 16 days, but not at 18 days. In both strains, there were more ANP cells in the left ventricle than in right beginning at day 13. There were no obvious strain differences in the developmental pattern and timing of ANP producing cells. However, on the day of birth, staining was reduced in hearts from some WKY newborn pups compared with hearts from SHR newborns and ventricular staining was reduced in both strains when compared to fetal hearts. These observations indicate that ANP is one of the earliest peptide hormones produced and that the predisposition to genetic hypertension does not appear to influence the development of ANP. PMID- 2972231 TI - Effects of B-aminopropionitrile on mineralization during endochondral ossification in chick tibia. AB - Two-week-old white leghorn chicks were fed a diet containing BAPN (0.05%) for three weeks. Thirty-six hours before sacrifice, the controls and BAPN fed chicks were dosed with 35S. The zone of provisional calcification was isolated, and 35S incorporation was estimated by liquid scintillation counting. Alkaline phosphatase and Ca+2-ATPase were biochemically analyzed. Microdensitometry, to assess the level of mineralization, was done on epiphysis and the metaphysis. Morphometry was performed on the various zones of growth plate. 35S incorporation was significantly lower in the bones of BAPN treated chicks as compared to the controls. The enzymatic studies showed a significant inhibition of alkaline phosphatase and Ca+2-ATPase. The microdensitometric studies showed a smaller area of highly mineralized bone in the zones of provisional calcification of the BAPN treated chicks as compared to the controls. Morphometry showed a reduction in the width of the zone of calcification in BAPN treated chicks as compared to the controls. On the basis of the above data, it is suggested that BAPN induced inhibition of mineralization during endochondral ossification may be the result of a lower synthesis of sulfur containing GAG's, the inhibition of enzymes alkaline phosphatase and Ca+2-ATPase and the derangement of cellular zones of the growth plate. The implications of these results lie in the fact that mineralization is dependent on multifactorial control of the microenvironment of bone and cartilage. PMID- 2972233 TI - Clinical pharmacology of doxacurium chloride. A new long-acting nondepolarizing muscle relaxant. AB - Doxacurium chloride (BW A938U) is a bis-quaternary benzylisoquinolinium diester nondepolarizing neuromuscular blocking compound that is minimally hydrolyzed by human plasma cholinesterase. The effect of bolus doses of doxacurium ranging from 10 to 80 micrograms/kg were studied in 81 consenting ASA physical status I and II patients anesthetized with nitrous oxide-oxygen-fentanyl-thiopental. The neuromuscular and cardiovascular effects of doxacurium were compared with those of eight patients receiving 100 micrograms/kg of pancuronium receiving identical anesthesia. The calculated ED95 for evoked twitch inhibition of the adductor pollicis at 0.15 Hz was 30 micrograms/kg. At 1.3 times the ED95 dose of doxacurium, recovery times to 5% and 25% of control twitch height were 59.2 +/- 4.1 (n = 23 of 26) and 75.7 +/- 5.6 (n = 23 of 26) min respectively. For pancuronium comparable recovery times were 81.7 +/- 10.3 (n = 8 of 8) and 83.0 +/ 8.4 (n = 5 of 8) min. Residual doxacurium blockade was readily antagonized by neostigmine. No dose-related effect on heart rate or mean arterial pressure was seen with doxacurium at doses up to and including 2.7 times the ED95 (80 micrograms/kg). Doxacurium administration did not result in any elevation of plasma histamine at doses up to and including 2.7 times the ED95. In this study doxacurium appears to be a long-acting nondepolarizing relaxant with readily reversible neuromuscular blocking effects and devoid of cardiovascular effects. This profile offers clinical advantages over current long-acting agents and further clinical trials seem appropriate. PMID- 2972232 TI - Reversible effects of long-term treatment with D-Trp6-LH-RH-microcapsules on pituitary-gonadal axis, spermatogenesis and prostate morphology in adolescent and adult dogs. AB - The effect of long term treatment with D-Trp6-LH-RH in microcapsules (GnRH-A) on pituitary gonadal axis was studied in a adolescent and adult Fox-terrier dogs. They received intramuscularly 50 micrograms/GnRH-A/kg, on day 1 and 21 and every 4 weeks thereafter. Three adult dogs received 4 injections. cLH, cFSH and T levels were undetectable on day 7. Detectable then normal levels occurred 60 and 90 days respectively after the last injection on day 77. Testis thickness was respectively 22.1 +/- 0.8 mm and 16.3 +/- 0.8 mm on days 0 and 77; initial values were observed 90 days later. Spermatozoa disappeared from the ejaculate on day 21 in 2 dogs; reappearance and complete recovery were observed on days 161-175 and 252 respectively. Histological findings showed on day 91 atrophic lesions of testis and prostate and spermatogonia were present in all seminiferous tubules. After recovery a normal histological appearance was noticed. Three adolescent 29 weeks old dogs received 14 injections, the last one on day 357. cLH, cFSH and T levels were undetectable only from day 105. Testis thickness were respectively 15.8 +/- 0.7 mm, 18.1 +/- 0.7 mm, 12.5 +/- 0.3 mm and 21.4 +/- 0.7 mm on day 0, 21, 357 and 490. Initially, no spermatozoa were present in the ejaculates, they appeared in 2 dogs after 2 months for 20 to 40 days then disappeared until day 449. Normal semen characteristics were observed in all three dogs on day 581. Histological findings on day 371 were comparable to those observed in the adult dogs. This study demonstrates that longterm treatment with D-Trp6-LH-RH in microcapsules leads to a reversible inhibition of spermatogenesis in dogs. The delayed response in adolescent dogs might be due to a transient resistance to therapy related to. PMID- 2972234 TI - The effects of succinylcholine on doxacurium-induced neuromuscular blockade. PMID- 2972235 TI - Hypotension and cutaneous reactions associated with intravenous administration of etoposide in the dog. AB - A study was undertaken to determine the pressor and toxic effects of etoposide, an antineoplastic agent, when administered IV in 0.9% sodium chloride solution (0.4 mg of etoposide/ml) over a 30-minute period to dogs at a dosage of 40 mg/m2 of body surface. On day 1, 6 adult German Shorthaired Pointers were anesthetized with halothane, and blood pressures were measured via a femoral artery catheter before, during, and after the etoposide was administered. Systolic, diastolic, and mean blood pressures of each dog decreased [corrected] significantly (P less than 0.01) within 30 minutes after initiation of etoposide infusion. On day 3, when the dogs were not anesthetized, etoposide was again administered to each dog, using the same dosage. Each dog developed a moderate to severe cutaneous reaction characterized by moderate to severe pruritus, urticaria, and swelling of the head and extremities that began during the second infusion of etoposide. These same cutaneous reactions were seen on day 30, when etoposide was administered to 3 of the previously treated dogs and 2 previously untreated Beagles. We concluded that the administration of the commercial preparation of etoposide is likely to cause a significant reduction in blood pressure of anesthetized dogs, and that the drug is likely to induce a moderate to severe cutaneous reaction when administered to unanesthetized dogs. PMID- 2972236 TI - Effects of zearalenone on various components of rabbit uterine tubal fluid. AB - A study was conducted to determine whether an orally administered mycotoxin, zearalenone (11.5 mg/kg of body weight/day), enters the uterine tubal fluid (UTF) of rabbits and to evaluate its influence on certain constituents of UTF. Using high-performance liquid chromatography, the concentration of zearalenone in UTF was maximal on day 1 of initial exposure to the compound, then decreased rapidly to half the peak concentration by day 5. The disappearance curve for zearalenone in UTF was linear throughout the treatment period (10 days) and was not detectable within 3 days after the last administration of zearalenone and the initiation of pregnancy (does were bred on day 10 of the zearalenone treatment). Zearalenone increased the volume of UTF (P less than 0.05) in estrous does, but decreased the pH (P less than 0.05) and the total free amino acid concentration (P less than 0.05) of UTF, compared with pretreatment values. The concentrations of several trace elements and amino acids known to be of critical importance in early embryonic development were altered by zearalenone treatment and/or pregnancy. Gross developmental anomalies were not evidenced when fetuses were examined on days 28 to 30 of gestation. It was concluded that zearalenone or associated metabolites affect factors that influence fertility during the early preimplantation period. Furthermore, the analysis of UTF appears to be an effective screening method for assessing certain potential reproductive toxicants. PMID- 2972237 TI - Use of febantel or ivermectin for treatment of calves with experimentally induced Bunostomum phlebotomum infection. AB - In the first of 2 separate trials, the efficacy of febantel, given at a dosage of 5 mg/kg of body weight, was assessed in calves with 60-day experimentally induced Bunostomum phlebotomum infection. Ten calves were given febantel paste, and 10 were given the vehicle only. All 20 calves were necropsied 7 days after cessation of treatment. Compared with untreated calves, febantel-treated calves harbored 99.4% fewer nematodes. In the second trial, the efficacy of ivermectin, given as a paste formulation at a dosage of 0.2 mg/kg, was assessed in calves with experimentally induced B phlebotomum infection. Ivermectin was given at 18 (n = 6) and 60 (n = 6) days after infection. At each treatment date, 3 additional calves were given vehicle only. At 67 days after infection, all calves were euthanatized. Efficacies of ivermectin against 18- and 60-day infections were 100 and 99.8%, respectively. Both anthelmintic preparations were easily administered, and adverse reactions were not observed. PMID- 2972238 TI - Rectus sheath hematomas: their pathogenesis, diagnosis, and management. AB - Rectus sheath hematomas are an uncommon entity causing an abdominal pain that may mimic other serious intraabdominal conditions. Knowledge of its predisposing factors and presentation are paramount in making the diagnosis. To facilitate the understanding of this condition, the authors reviewed their experience at St. Joseph Mercy Hospital in Ann Arbor, Michigan for the period between 1977 and 1986. Eight patients who were hospitalized for other reasons were found to have rectus sheath hematomas. All patients were women with an age range of 26 to 85 years and a mean age of 58 years. The correct diagnosis was made clinically in four patients. In those in whom hematomas of the rectus sheath were not initially diagnosed, surgery was undertaken, at which time the condition was identified. Predisposing factors include trauma, childbirth, previous operation of the lower abdomen, and anticoagulant use. Ultrasonography may aid in the diagnosis but CT scan is most accurate in its ability to define the lesion. When diagnosed clinically, a conservative therapeutic program can usually be instituted. PMID- 2972239 TI - Influence of cyclosporine on CTL behavior in vitro and in vivo. PMID- 2972240 TI - Soluble antigen-specific helper molecules active in the induction of cytotoxic T lymphocytes. PMID- 2972241 TI - Cooperation between helper T cells and cytotoxic T lymphocyte precursors. AB - We have investigated the requirement for the presence of L3T4-positive T cells in the in vivo priming of Lyt-2-positive cytotoxic T-lymphocyte precursors. The antigens used were the male-specific antigen H-Y and autosomal minor histocompatibility antigens. In some experiments, responder mice were depleted of L3T4-positive cells by repeated intraperitoneal injections of the anti-L3T4 antibody, GK1.5. In other experiments, lymphoid cells from normal mice were fractionated in vitro by antibody-plus-complement treatment and the populations primed in irradiated adoptive hosts. In these antigen systems, depletion of L3T4 positive helper cells decreases the level of priming of cytotoxic T lymphocytes. With regeneration of the L3T4-positive subpopulation, the CTL response to antigen increases. To some extent, the reliance on L3T4-positive cells can be overcome by increasing the antigen dose. We conclude that in most physiological responses, L3T4-positive T cells enhance the cytotoxic T-cell response. PMID- 2972242 TI - The veto phenomenon in T-cell regulation. PMID- 2972243 TI - Function of granule perforin and esterases in T cell-mediated reactions. Components required for delivery of molecules to target cells. AB - Cognate T cell-mediated functions require antigen and MHC-restricted recognition of target cells. T-effector functions comprise the delivery of signals for help, for suppression, or for cell death of the target cell. In the case of the delivery of cytotoxicity and of help for B-cell antibody production, it is known that the secretory apparatus of the effector cell participates. Prior to secretion, many components of the effector cell are stored in cytoplasmic granules. Among the important and apparently constant constituents of granules are pore-forming proteins (perforins) and proteinases (granzymes). The putative role of perforin has been thought to mediate direct cytotoxicity. It is postulated here that, in addition, perforin at low concentrations may induce target-cell endocytosis through the formation of Ca channels. Localized endocytosis of the target at the contact site in turn may lead to the uptake of locally secreted effector-cell factors, such as cytotoxic factors (CTL), lymphokines (helper cells), or suppressor factors (suppressor cells). The potential importance of such a mechanism is the delivery and uptake of secreted effector-cell components into the endosomes of target cells, bypassing the need for appropriate target-cell receptors. Perforin thus may subserve two functions depending on its intragranular concentration: one, as a killer molecule, and two, as a delivery system for additional granule factors. One of the roles of esterases in T cell-mediated cognate-effector functions may be to allow recycling of the effector cell. This apparently is achieved by an active process of detachment of the effector T cell from the target cell, possibly by way of the proteolytic cleavage of adhesion molecules. Esterases are secreted, together with perforin and other factors, during granule release at the effector target-contact site, where they can cleave intercellular adhesion molecules and thus allow effector-cell recycling and attachment to new target cells. Other roles of esterases, not discussed here, may include participation directly in the cytotoxic process through uptake into the target cell. The evidence for a common intercellular molecular delivery mechanism of cognate effector T-cell function involving perforin and esterases is summarized. This concept represents a unifying hypothesis for MHC-restricted, contact-requiring, intercellular T cell signal delivery as well as for the delivery of cytotoxicity by non-MHC-restricted T cells and natural killer cells. PMID- 2972244 TI - Tumor-infiltrating lymphocytes in patients with renal-cell carcinoma. PMID- 2972245 TI - The Mls reaction. A regulatory phenomenon. PMID- 2972246 TI - Cellular interactions resulting in skin-allograft rejection. PMID- 2972247 TI - Donor-directed cytotoxic T cells and other inflammatory components of acute allograft rejection. PMID- 2972248 TI - Steroid hormone receptors and response to high-dose medroxyprogesterone acetate in advanced breast cancer. AB - Forty-six patients with advanced breast cancer were treated with oral high-dose medroxyprogesterone acetate (MPA) as a second line endocrine treatment, with a 28% 13/46) response rate. There was a significant difference in response to MPA between patients with estrogen (ER)- or progesterone (PgR)-receptors and those with negative receptors assayed just before the treatment (ER+, 7/10, ER-, 0/9 of response rates, p = 0.007, PgR+, 4/4: PgR-, 3/15, p = 0.02). The combination of ER and PgR was observed to be best in predicting the response (p = 0.02). The ER status as well as the combination of ER and PgR, but not PgR alone detected in the primary tumors, correlated well with the response (ER+, 10/23: ER-, 0/15, p = 0.0094, PgR+: PgR-, p = 0.13, ER+PgR+, ER+PgR-, ER-PgR-:p = 0.024). The correlation between steroid hormone receptors and response to MPA in advanced breast cancer was established from the literature. Response rate to MPA was shown to be 42% (69/169) in ER+ cancer patients, and 10% (9/90) in ER- cancer patients (p = 0.00004). There was no correlation between PgR status and the response. Changes of receptors were studied in relation to the response. When ER+ or PgR+ tumors retained their positivity until MPA treatment, a good response was obtained, while there were almost no responders if one or both receptors changed from positive to negative, or remained negative. In conclusion, the response to MPA can be predicted by the steroid hormone receptors. PMID- 2972249 TI - Loss of functional suppression is linked to decreases in circulating suppressor inducer (CD4+ 2H4+) T cells in multiple sclerosis. AB - A consistent immunological finding in patients with progressive multiple sclerosis is a loss of functional suppression. We have recently found decreases in suppressor inducer T cells in progressive multiple sclerosis as measured by two-color immunofluorescence using differentiation markers CD4 and 2H4. In the present study, we examined the relationship between functional suppression and circulating CD4+ 2H4+ T cells using a two-stage assay. (1) T cells were stimulated for 7 days with irradiated non-T cells (autologous mixed lymphocyte reaction [AMLR]) and harvested. It has previously been shown that suppressor T cells are generated during the course of the AMLR. (2) The AMLR-generated suppressor T cells were then incubated with mononuclear cells plus pokeweed mitogen, and immunoglobulin (Ig) synthesis was measured. There was less AMLR induced suppression of IgG synthesis in patients with progressive multiple sclerosis as compared with normal subjects and patients with other neurological diseases. More importantly, there were significant correlations between decreases in circulating CD4+ 2H4+ cells and the AMLR (p = 0.009). Thus, the decreases in functional suppression and the decreases in the AMLR in multiple sclerosis appear tightly linked to CD4+ 2H4+ cells, and their measurement provides a means to monitor suppressor function phenotypically. Decreases in suppressor inducer T cells may in part explain immunoregulatory abnormalities observed in multiple sclerosis. PMID- 2972250 TI - Fluctuations of CD4+ T-cell subsets in remitting-relapsing multiple sclerosis. AB - Patients with multiple sclerosis (MS) frequently have selective depletion of the CD45R+CD4+ T-cell subset during active phases of disease. To study the relationship between changes in this subset and the onset of objective clinical exacerbations of disease, a longitudinal study was undertaken. Two CD4+ T-cell subsets and two CD8+ T-cell subsets were monitored by two-color immunofluorescence using a fluorescence-activated cell sorter. These subsets of peripheral blood lymphocytes were monitored monthly for one year in a group of 9 patients with remitting-relapsing MS and in 11 healthy age-matched control subjects. Significant changes in the ratio of two CD4+ T-cell subsets (CD45R /CD45R+) were detected in 7 of 9 patients with MS, but not in any of the control subjects. Of those 7 persons, 4 suffered major clinical relapses substantiated by alterations in the neurological examination. The other 3 suffered minor relapses with subjective clinical abnormalities. All 7 had increased CD4+ T-cell subset ratios (%CD4+CD45R-/%CD4+CD45R+) within the month that new symptoms were reported. Most such increases resulted from a simultaneous depletion in the number of CD45R+CD4+ T cells and an increase in the number of CD45R-CD4+ T cells. One patient suffered a major relapse with no change in the ratio of CD4+ subsets but had a depletion of all CD4+ T cells. There were no consistent changes in any of the other subsets measured. These results indicate that a subgroup of patients with MS have abnormal fluctuations of two CD4+ T-cell subsets, which may correlate with increased disease activity. PMID- 2972251 TI - Cerebrospinal fluid homovanillic acid and parkinsonism in Huntington's disease. AB - In Huntington's disease (HD), normal or decreased levels of homovanillic acid (HVA) in cerebrospinal fluid (CSF) have been reported but have not been analyzed with respect to severity of parkinsonism, which in certain cases may be a predominant feature of the illness. We obtained CSF by lumbar puncture from four groups of nonmedicated subjects: (1) those with HD in the earliest stages of illness (n = 51), (2) those with parkinsonism, including idiopathic (n = 10) and atypical forms (n = 4), (3) those with nonparkinsonian movement disorders (n = 19), and (4) normal volunteers (n = 4). HVA was determined by high-pressure liquid chromatography with electrochemical detection, and motor signs were assessed in standardized fashion. The parkinsonian group had reduced levels of CSF HVA, but the other groups showed no significant differences. For those with HD, no correlation was found between HVA level and severity of parkinsonism, and there were no differences in HVA level between those subjects with (n = 14) or without (n = 37) prominent parkinsonism or between subjects whose age at illness onset was 30 years or less (n = 16) and those whose age at onset was over 30 (n = 35). Our findings indicate that in early, untreated HD, CSF HVA is in the normal range and does not correlate with the severity of parkinsonism. This observation supports neuropathological findings suggesting that parkinsonian features in HD are largely related to the loss of postsynaptic striatal dopamine receptors rather than to presynaptic nigral degeneration. PMID- 2972252 TI - Fibronectin and its receptors. PMID- 2972253 TI - Mycotoxins and Fusarium spp. associated with infected ears of corn in Minnesota. AB - Five Fusarium species were isolated from the grain of dent corn (Zea mays) selected from 20 of 32 damaged fields in 10 counties in Minnesota on the basis of hyphal growth visible on kernels in the field. Three mycotoxins were identified in the infected ears: zearalenone, deoxynivalenol, and 15-acetyl-deoxynivalenol. This is the first report of the presence of 15-acetyl-deoxynivalenol on corn ears in the field prior to harvest and in combination with deoxynivalenol and zearalenone. Ninety-nine cultures were selected from colonies growing from kernels on an agar medium; 30% of the cultures were F. graminearum, 23% were F. subglutinans, 20% were F. moniliforme, 14% were F. oxysporum, and 12% were F. proliferatum. PMID- 2972255 TI - MgATP-dependent, glucose 6-phosphate-stimulated liver microsomal Ca2+ accumulation: difference between rough and smooth microsomes. AB - Some features of the MgATP-dependent Ca2+-accumulating capacity of rough as compared to smooth liver microsomal fraction were studied. Smooth microsomes accumulate somewhat higher amounts of Ca2+ than rough ones in the presence of MgATP. In the presence of glucose 6-phosphate, which markedly stimulates MgATP dependent Ca2+ accumulation in both fractions, smooth microsomes exhibit a much higher Ca2+-accumulating capacity than rough ones. Possible reasons of the differences observed between the two fractions were investigated. Smooth microsomes exhibit a higher Ca2+-dependent ATPase activity, suggesting a higher Ca2+ inward transport into smooth vesicles. Also, following the inhibition of active Ca2+ transport by means of vanadate, smooth microsomes appear to release the Ca2+ previously accumulated--both in the absence (i.e., with MgATP only) and in the presence of glucose 6-phosphate--at a lower rate than rough ones. This indicates a lower passive backflux of Ca2+ accumulated in smooth vesicles. On the basis of these data, differences can be envisaged with respect to cellular Ca2+ handling by different domains of endoplasmic reticulum in the liver. PMID- 2972254 TI - Formation of fusarenone X, nivalenol, zearalenone, alpha-trans-zearalenol, beta trans-zearalenol, and fusarin C by Fusarium crookwellense. AB - Fusarium crookwellense KF748 (NRRL A-28100) (isolated from dry rotted potato tubers in Central Poland) produced six mycotoxins on both rice and corn substrates at 25 degrees C. The metabolites detected were zearalenone, alpha trans-zearalenol, beta-trans-zearalenol, fusarin C, and the trichothecenes fusarenone X and nivalenol. This is the first report of formation of alpha-trans zearalenol, beta-trans-zearalenol, fusarenone X, and nivalenol by F. crookwellense. PMID- 2972257 TI - Characterization of nicked myotoxin a and its effect on the sarcoplasmic reticulum calcium pump. AB - Myotoxin a, a muscle-necrotizing polypeptide isolated from Crotalus viridis viridis (prairie rattlesnake) venom, was nicked at Met-28 by cyanogen bromide. Amino acid analysis indicated that the methionine content was reduced to zero from the original 1 mol. Judging from circular dichroism, the nicked myotoxin a had a conformation similar to that of original myotoxin. Raman spectra indicated that the conformations of the three disulfide bonds are not affected in nicked myotoxin a. Like the original toxin, nicked myotoxin a was myotoxic and inhibited calcium ion loading activity, although the inhibitory action was slightly lower than that of the original myotoxin a. Both modified and unmodified myotoxin a showed myonecrotic activity as determined by examining histological slides. The modified toxin also inhibited the formation of decavanadate-induced two dimensional crystalline arrays of the sarcoplasmic reticulum Ca2+-ATPase just as the original myotoxin a does. PMID- 2972256 TI - Motility, heat, and lactate production in ejaculated bovine sperm. AB - Effects of various inhibitors on motility, heat, and lactate production of ejaculated bovine sperm were determined in the presence of antimycin A and rotenone. erythro-9-[3-(2-Hydroxynonyl)]adenine (EHNA) and polyvinylpyrrolidone (PVP-360) stopped motility and reduced heat or lactate production by 30-50%. Carbodiimides resulted in loss of motility and a reduction of metabolism by 60 75%. Quercetin treatment, which enhanced rather than inhibited motility, depressed heat and lactate production by 50-60%. Since mechanical immobilization reduced heat production by only 30%, the question arises as to what other cellular processes are major contributors to the energy budget. Inhibitors of ion flux had little-to-no effect on heat or lactate production, suggesting that neither mitochondrial nor Na+/K+ ATPases were major ATP-requiring processes. Calcium flux at the plasma membrane also was minimal and previous reports eliminated glycolytic substrate cycling as major consuming processes for ATP. Although quercetin inhibited lactate production in intact cells, no effect of quercetin on cell-free glycolysis and the ATPase activities of isolated dynein was detected. Quercetin did, however, inhibit ATPase activity of plasma membrane, suggesting that this unidentified ATPase may contribute to the formation of ADP and Pi required for lactate production by the intact cell. We propose (a) that the bioenergetic costs of motility are divided between regulatory events and dynein-microtubule interaction (dynein ATPase), (b) that some of the membrane related processes may be "inefficient," and (c) that quercetin may render these steps more "efficient," in a manner analogous to its action on the Na+/K+ pump of Ehrlich ascites tumor cells. PMID- 2972258 TI - [Clinical trial of 5'-DFUR in patients with recurrent breast cancer]. AB - 5'-DFUR was administered orally to recurrent breast cancer patients at a daily dosage of 1,200 mg given 3 times a day for more than 8 weeks. Out of 16 evaluable cases, 1 CR, 5 PR, 5 NC and 5 PD were observed, and the overall response rate was 37.5%. There was no significant difference in the response rate between the patient with or without prior fluorinated pyrimidine therapy, or between sites of the lesion. Toxic effects consisted of gastrointestinal toxicity such as diarrhea (25%), anorexia (12.5%), abdominal pain (12.5%) and nausea and vomiting (6.3%). No other severe side effect was observed. These results suggest that 5'-DFUR can be useful for the treatment of breast cancer. PMID- 2972259 TI - Topical antibiotics in dermatology. AB - Topical antibiotics are safe and effective in certain conditions, primarily acne, rosacea, and nasal carriage of Staphylococcus aureus. They are useful in impetigo only when it is of limited extent. Their efficacy in other pyodermas is unclear, although mupirocin is probably effective in many cases. In "infected eczema" that does not require systemic therapy they seem to add little to what topical corticosteroids alone achieve. They are ineffective in reducing the incidence of significant infection with indwelling intravenous catheters. They are safe preparations, but extensive use, especially in closed populations, may encourage the emergence of resistant bacteria. PMID- 2972260 TI - Intraspinal tumours. PMID- 2972261 TI - Distribution of Langerhans cells in skin lesions of MRL/l mice. PMID- 2972263 TI - Epidemiology and vectors of Dracunculus medinensis in northwest Burkina Faso, West Africa. AB - The epidemiology of the guinea worm, Dracunculus medinensis, was studied in the dry savanna zone of West Africa. The monthly incidence data collected over a period of four years showed peak transmission occurring in June and July at the beginning of the rainy season. The different types of local water sources (i.e. wells, periodic streams, seasonal cattle waterings, natural ponds and man-made ponds) were examined for infected cyclopoid copepods. Small hand-dug ponds or water-holes situated in the fields proved to be the most important sites of transmission. While the domestic water supply is obtained from draw-wells in the villages throughout the year, the villagers take additional drinking water from these ponds during the planting season when farm activities require long stays in the fields. Four cyclopoid species were found for the first time acting as natural intermediate hosts of D. medinensis. Thermocyclops inopinus was the most frequently infected cyclopoid, and small man-made ponds are the preferred habitats of this species. Occurrence of T. inopinus is confined to the first half of the rainy season, coinciding with peak transmission. The epidemiology of dracunculiasis in dry and humid regions of West Africa is compared with regard to seasonality. The use of protective water filters proved to be the only adequate method for guinea worm control in the project area. PMID- 2972262 TI - Source of drinking water supply and transmission of guinea worm disease in Nigeria. AB - During 1982, ecological factors associated with freshwater pollution were investigated in Idere, a rural Nigerian community with endemic guinea worm infection. Data were collected on the quality of all pond water sources, and on rainfall patterns and potable piped water available in the community. Pond water provided 76% of the total water used in Idere. This source of drinking water provided the classical ecological environment for the transmission of Dracunculus medinensis, other helminth parasites and bacterial enteric infections. The bacteriological analysis of drinking water from the ponds reflects the absence of sanitary arrangements for human waste disposal in the community, as the ponds are collectors of storm run-offs. Okina, the spring-fed pond which was nearest to the households, was the most reliable year-round source of water to the community; however, Okina also contained the highest density of infective Thermocyclops and the highest faecal coliform (FC) to faecal Streptococcus (FS) ratio (FC/FS), thus providing a central reservoir for guinea worm and bacterial infections. The transmission season of guinea worm infection corresponded with the period of greatest water scarcity in Idere. The amount of portable water available to Idere residents in 1981 was 3.6 litres per person per day. Frequent mechanical breakdowns, electric power failures, lack of fuel to run the water pumping engines and the direct link of the water pipeline supplying water to Idere with a water pipeline serving another major city in the same district were some of the reasons for potable water shortage in the community. PMID- 2972264 TI - Metabolism in Bunostomum trigonocephalum. Sixth communication: lipid biosynthesis from C14-labelled acetate and glucose in the two sexes and third-stage infective larvae. PMID- 2972265 TI - Early parental loss and development of adult psychopathology. AB - We assessed the effect of parental loss during childhood on the development of psychopathology in 90 adults. The subjects with a history of adult psychopathology (PATH group), in comparison with subjects with no history of a psychiatric disorder (NO PATH group), had poorer quality of childhood home life and personal adaptation subsequent to parental loss as assessed by the Home Life and Personal Adaptation (HAPA) scale developed by us. Total HAPA scale scores were the single most powerful predictor of adult psychopathology, accounting for correct prediction of adult psychopathology in 80% (72/90) of the subjects. The PATH subjects had significantly increased plasma levels of cortisol and beta endorphin immunoreactivity. Moreover, cortisol and adrenocorticotropic hormone levels significantly correlated with total HAPA scores. First-degree family history of psychiatric disorders, age at loss, and parental vs maternal loss were not significantly different between PATH and NO PATH subjects. We conclude that the quality of home life subsequent to early parental loss is critically related to the development of adult psychopathology. The hypothesis that early trauma results in enduring neuroendocrine alterations in hypothalamic-pituitary-adrenal axis function is examined. PMID- 2972266 TI - Lipoprotein(a). A potential bridge between the fields of atherosclerosis and thrombosis. AB - Lipoprotein(a) (Lp[a]) represents a class of plasma lipoprotein particles that have overall characteristics similar to low-density lipoproteins but distinct from them by having apolipoprotein B100 linked to apolipoprotein(a) by disulfide bridge(s). This protein has recently been shown to have a striking amino acid sequence homology with plasminogen, a serine protease zymogen that on activation to plasmin promotes the conversion of fibrinogen to fibrin. The high incidence of Lp(a) in the plasma of patients with cardiovascular disease has been noted by many investigators. The new knowledge being rapidly acquired on the structure of Lp(a) should facilitate the understanding of the mechanism of its atherogenicity and perhaps shed light on its possible physiologic role. PMID- 2972267 TI - Protein S deficiency. PMID- 2972268 TI - Low back disorders: conservative management. AB - Painful disorders of the low back may arise from a large variety of mechanisms and there is no simple, single solution to them. They have a great social and economic cost in industrialized countries. The major syndromes of low back pain and sciatica are reviewed, including myofascial pain syndromes, disc herniations, radiculopathies, spinal stenosis and facet syndromes. Non-operative management is reviewed, including bed rest, medications, traction, manipulation, external supports, physical modalities, therapeutic exercise, trigger point injection and chemonucleolysis. Prevention should become the main goal of physicians dealing with this multifaceted problem. PMID- 2972269 TI - Vascular complications of percutaneous femoral cardiac interventions. Incidence and operative repair. AB - Of 7333 patients undergoing percutaneous left-heart catheterization procedures during a seven-year seven-month period, 73 patients (1%) underwent 75 operative repairs of catheterization-related vascular complications. The overall incidence of operative repair varied according to the type of percutaneous femoral artery procedure performed: 0.6% for diagnostic heart catheterization, 0.9% for percutaneous transluminal angioplasty, 5.2% for transfemoral balloon valvuloplasty, and 11.5% for intra-aortic balloon pump placement. This suggests that (1) femoral and iliac artery occlusions can and should be repaired promptly while the patient is under local anesthesia; (2) false aneurysms in this clinical setting can be approached directly through the aneurysm cavity; (3) the diagnosis of false aneurysm or arteriovenous fistula can usually be made on clinical grounds alone, without resorting to angiography; (4) venous or arterial patch angioplasty is the preferred technique for small or severely traumatized femoral arteries; and (5) the necessity for operative repair of these lesions will continue to increase in frequency as percutaneous cardiologic diagnostic and therapeutic interventions are used more widely. PMID- 2972270 TI - Amantadine, fatigue, and multiple sclerosis. AB - In a double-blind placebo-controlled crossover study of ten patients with multiple sclerosis, we found amantadine hydrochloride therapy to be effective in improving fatigability in six. Administration of the drug was associated with significantly higher levels of beta-endorphin-beta-lipotropin and responders had significantly higher levels than nonresponders. Lactate levels were significantly higher and pyruvate levels lower in nonresponders. Amantadine given for fatigue to patients with multiple sclerosis is associated with measurable changes in levels of metabolites and peptides in the circulation. PMID- 2972271 TI - Immunogenicity study of a plasma-derived heat-inactivated hepatitis-B vaccine in newborn babies: a randomised dose-response study. PMID- 2972272 TI - [Possibilities for simultaneous determination of ABO, PGM1 and Gc polymorphisms in typing of blood stains]. PMID- 2972273 TI - Identification and partial purification of ABGP205, an integral membrane glycoprotein from brain that binds ankyrin. AB - 1. A procedure was devised that allows the membrane-skeletal proteins brain spectrin and ankyrin to be extracted selectively from a membrane-skeletal preparation, together with some actin, an Mr-103,000 protein and a population of glycoproteins. 2. Ankyrin-binding activities of the glycoproteins were investigated by affinity chromatography. We detected only one, Mr 205,000, that binds ankyrin and is prevented from binding by the cytoplasmic domain of Band 3, the established erythrocyte-membrane-binding site for ankyrin. The Mr-205,000 glycoprotein, designated ABGP205, may be a candidate for a membrane-binding site for ankyrin. PMID- 2972274 TI - Two populations of complement factor H differ in their ability to bind to cell surfaces. AB - Using hydrophobic affinity chromatography on phenyl-Sepharose, human complement factor H can be separated into two subpopulations, phi 1 and phi 2. Although phi 1 and phi 2 are known to differ in their aggregation properties under non physiological low ionic strength conditions, no difference in aggregation state was detected under the conditions used for cell-binding experiments. We have investigated these two subpopulations further to determine whether functional differences exist between them. The subpopulation phi 2 was found to bind specifically and saturably to the surface of Raji cells. The binding of the other subpopulation, phi 1, was low, and essentially non-specific. A monoclonal anti factor H antibody, BGH-1, was raised which recognizes preferentially the phi 2 subpopulation and inhibits the binding of factor H to cell surfaces. PMID- 2972275 TI - Identification of specific binding sites for keratan sulphate proteoglycans and chondroitin-dermatan sulphate proteoglycans on collagen fibrils in cornea by the use of cupromeronic blue in 'critical-electrolyte-concentration' techniques. AB - Proteoglycans (PGs) in bovine corneal stroma were stained with Cupromeronic Blue in 'critical-electrolyte-concentration' (CEC) methods for electron microscopy, and were located vis-a-vis collagen fibril a-e banding patterns. Keratanase and chondroitin ABC lyase digestion showed that a + c-band- and d + e-band-associated PGs were keratan sulphate-rich and chondroitin (dermatan) sulphate-rich respectively. The CEC pattern proved that the keratan sulphate PGs at the a and c bands differed. Comparison of their CECs with their behaviour on anion-exchange chromatography confirmed previous (indirect) attempts at identification [Scott & Haigh (1985) Biosci. Rep. 5, 765-774]. Similar arguments were applied to the dermatan sulphate PGs at the d and e bands. These results strongly support the one-PG-one-binding-site hypothesis [e.g. Scott (1988) Biochem. J. 252, 313-323]. Remarkable inter-species variations in the keratan sulphate PG patterns contrast with the relatively constant picture of dermatan sulphate PG-collagen fibril interactions. PMID- 2972277 TI - Binding of fructose 2,6-bisphosphate to yeast phosphofructokinase. AB - Binding of Fru-2,6-P2 to yeast phosphofructokinase was investigated by ultrafiltration technique. Per mol of subunit of phosphofructokinase (M = 100,000) 0.5 moles of Fru-2,6-P2 are bound. The binding curve proceeds cooperatively (nH = 1.8 +/- 0.2). The apparent affinity constant of Fru-2,6-P2 amounts to about 2.25 +/- 0.12 microM. Fru-1,6-P2 decreases the affinity of yeast phosphofructokinase to Fru-2,6-P2. The data can be described by assuming either competition of Fru-2,6-P2 and Fru-1,6-P2 for the same binding site or conformationally mediated interactions. PMID- 2972276 TI - Hydrolysis of alpha-human atrial natriuretic peptide in vitro by human kidney membranes and purified endopeptidase-24.11. Evidence for a novel cleavage site. AB - alpha-Human atrial natriuretic peptide (hANP) is secreted by the heart and acts on the kidney to promote a strong diuresis and natriuresis. In vivo it has been shown to be catabolized partly by the kidney. Crude microvillar membranes of human kidney degrade 125I-ANP at several internal bonds generating metabolites among which the C-terminal fragments were identified. Formation of the C-terminal tripeptide was blocked by phosphoramidon, indicating the involvement of endopeptidase-24.11 in this cleavage. Subsequent cleavages by aminopeptidase(s) yielded the C-terminal dipeptide and free tyrosine. Using purified endopeptidase 24.11, we identified seven sites of hydrolysis in unlabelled alpha-hANP: the bonds Arg-4-Ser-5, Cys-7-Phe-8, Arg-11-Met-12, Arg-14-Ile-15, Gly-16-Ala-17, Gly 20-Leu-21 and Ser-25-Phe-26. However, the bonds Gly-16-Ala-17 and Arg-4-Ser-5 did not fulfil the known specificity requirements of the enzyme. Cleavage at the Gly 16-Ala-17 bond was previously observed by Stephenson & Kenny [(1987) Biochem. J. 243, 183-187], but this is the first report of an Arg-Ser bond cleavage by this enzyme. Initial attack of alpha-hANP by endopeptidase-24.11 took place at a bond within the disulphide-linked loop and produced a peptide having the same amino acid composition as intact ANP. The bond cleaved in this metabolite was determined as the Cys-7-Phe-8 bond. Determination of all the bonds cleaved in alpha-hANP by endopeptidase-24.11 should prove useful for the design of more stable analogues, which could have therapeutic uses in hypertension. PMID- 2972278 TI - Identification of a 29-amino acid natriuretic peptide in chicken heart. AB - Morphological and pharmacological observations have suggested that chicken atrial natriuretic peptide (ANP) is different from mammalian ANP. The present survey for the as yet unidentified ANP in chicken heart was performed by monitoring the relaxant effect on chick rectum. From the low molecular weight component of rectum relaxant activity observed in acid extracts of chicken ventricle, a novel 29-amino acid peptide was purified. The identical peptide was also isolated from acid extracts of chicken atrium. The peptide elicited a pharmacological spectrum very similar to that of mammalian ANP, including diuretic-natriuretic and hypotensive activity. Thus, the peptide was designated "chicken alpha-ANP (alpha chANP)". The complete amino acid sequence determined for the peptide showed remarkable homology with that of mammalian alpha-ANP. However, maximum homology was observed when the peptide was compared with a recently identified porcine brain natriuretic peptide (BNP). PMID- 2972279 TI - Identification of new atrial natriuretic peptides in frog heart. AB - It has been observed that mammalian atrial natriuretic peptide (ANP)-like immunoreactivity is found in frog heart, but to date the natriuretic factors have not yet been identified. Isolation from bull-frog heart extract was performed mainly by immunoaffinity chromatography on a column linked with anti-hANP IgG. From the low molecular weight fraction, 24- and 21-amino acid peptides were purified to homogeneity. Both peptides were found to elicit diuretic-natriuretic as well as vasorelaxant activity, and were named "frog ANP-24" and "frog ANP-21", respectively. Complete amino acid sequences of the peptides were established by microsequencing and confirmed by syntheses. Frog ANP-21 was identified as an N terminally three amino acid deleted form of frog ANP-24. Remarkable sequence homology was observed between frog ANP and mammalian ANP, especially in the regions flanked by two half-cystine residues. PMID- 2972281 TI - Physiological concentrations of atrial natriuretic factor stimulate human erythroid progenitors in vitro. AB - Human alpha-ANF[1-28] potentiated erythroid colony formation up to four-fold in cultures containing erythropoietin. Both early and late erythroid precursor cells responded to alpha-ANF[1-28] [0.032 to 1 nM] in a dose dependent fashion. Removal of T lymphocytes and macrophages which have been shown to modulate erythropoiesis did not abolish the stimulatory effect. All major circulatory forms of ANF (alpha ANF[1-28], alpha-ANF[4-28] and alpha-ANF[5-28]) had potent erythropoietic activity. These results indicate that concentrations of ANF reached during hypoxia stimulate erythroid progenitor cells in the presence of erythropoietin. PMID- 2972280 TI - Calcitonin gene-related peptide(CGRP) stimulates the release of atrial natriuretic peptide(ANP) from isolated rat atria. AB - The effect of calcitonin gene-related peptide(CGRP) on the release of atrial natriuretic peptide(ANP) was studied in spontaneously beating, isolated rat atria. CGRP stimulated the ANP release in a dose-dependent manner. When the atria were incubated with a combination of phentolamine, propranolol, and atropine, these antagonists blocked neither the rise in ANP release nor the positive chronotropic and inotropic effects of CGRP. Therefore, we conclude that CGRP stimulates ANP release as well as cardiac contractility independently of adrenergic and cholinergic receptors. PMID- 2972282 TI - Calcimedin, calelectrin: correlation of relatedness. AB - A statistical method has been used to compare the amino acid compositions of several calelectrin proteins and the 67k calcimedin protein. The validity of the method for determining protein relatedness and ancestry has been established with many different proteins. Torpedo calelectrin, p68 brain calelectrin, 67,000 dalton brain calelectrin, 67,000 bovine aorta protein, 32k lipocortin (and probably 67R lung protein) share considerable homology with each other and with calpactin I or lipocortin II, the pp60src tyrosine kinase substrate. The 67k calcimedin appears to be unrelated to these several proteins, in agreement with biochemical evidence, although limited homology may still be present. The lung 67E protein, calregulin and calmodulin, are unrelated to any of the other proteins. PMID- 2972283 TI - Growth hormone-dependent insulin-like growth factor (IGF) binding protein both inhibits and potentiates IGF-I-stimulated DNA synthesis in human skin fibroblasts. AB - This study investigates the effects of BP-53, the acid-stable IGF-binding subunit of the circulating 150 kDa IGF-binding protein complex, on IGF-I-stimulated thymidine incorporation by neonatal human skin fibroblasts. When cells were incubated for 24 h with IGF-I in serum-free medium, and thymidine incorporation studied over the final 4-h period, maximal stimulation (4- to 7-fold) was seen with 30 ng/ml IGF-I, with a half-maximal effect at 6.8 +/- 1.2 ng/ml (SD, n = 4). Co-incubation of IGF-I with increasing concentrations of pure BP-53 caused dose dependent inhibition of IGF-I-stimulated thymidine incorporation, which was complete when IGF-I and BP-53 were equimolar. In contrast, preincubation of cells with BP-53 for 8-48 h before adding IGF-I resulted in a potentiation of the subsequent IGF-I effect. The potentiation was maximal (2-fold) at a BP-53 concentration of 150 ng/ml, and appeared to act by increasing the maximal rate of thymidine incorporation rather than the sensitivity of this process to IGF-I. Since neonatal fibroblasts produce a protein which is identical to BP-53 in size and immunoreactivity, these results suggest an autocrine role for BP-53 in regulating fibroblast responsiveness to IGF-I. PMID- 2972284 TI - Physicochemical characterization of [3H] DHEA binding in rat liver. AB - Dehydroepiandrosterone (DHEA), the native clinical steroid and steroid precursor, may have a targeted physiologic role. A high affinity (Kd 2.3nM) and steroid specific [3H] DHEA binding macromolecule in male Sprague-Dawley rat hepatic cytosol suggests that DHEA may have receptor mediated physiologic action. 3[H] DHEA binding was highest in the liver followed by kidney and testis cytosols. Sulfhydryl reagents such as N-ethylmalemide and iodoacetamide inhibited the binding of [3H]DHEA by up to 60-70%. The DHEA-macromolecular complex was stable at 35 degrees C. and addition of 5mM molybdate or 0.3M KCl increased stability. Interestingly, rat liver cytosol, specific binding at 4 degrees C increased by almost 40-50% with addition of 0.1M NaSCN or 0.3M KCl. Sucrose gradient analyses showed a 7-8 S macromolecular complex in the low salt and 3-4 S complex under high salt conditions. The [3H] DHEA- macromolecular complex shows minimal temperature dependent activation in vitro at 25 degrees C as judged by binding to DNA-cellulose. The results suggest a specific high affinity macromolecule for DHEA in rat liver cytosol with unique physicochemical properties. PMID- 2972285 TI - Water immersion increases the concentration of the immunoreactive N-terminal fragment of proatrial natriuretic factor in human plasma. AB - Atrial natriuretic factor (ANF) N-terminal (ANF 1-98) and C-terminal (ANF 99-126) fragments were determined by radioimmunoassay in human plasma. Mean basal plasma ANF N-terminal concentrations in 9 healthy subjects were 461 +/- 58 fmol/ml, significantly (p less than 0.0001) higher than ANF C-terminal concentrations (4.8 +/- 0.5 fmol/ml). Central volume stimulation by one hour head-out water immersion (WI) induced a significant (p less than 0.01) increase of the C-terminal peptide levels to 11.6 +/- 2.3 fmol/ml, paralleled by a significant (p less than 0.001) increase of the N-terminal fragment levels to 749 +/- 96 fmol/ml. Increases of plasma concentrations of both fragments upon WI correlated significantly (r = 0.71; p less than 0.05). These data suggest cosecretion of the N-terminal fragment with the C-terminal fragment of pro ANF 1-126 following a physiological stimulus of ANF release in man. PMID- 2972286 TI - Abscisic acid enhances aggregation and fusion of phospholipid vesicles. AB - The plant hormone abscisic acid (ABA) is shown to enhance the aggregation and fusion of small unilamellar lipid vesicles composed of 80 mol% dimyristoylphosphatidylcholine (DMPC) and 20 mol% dimyristoylphosphatidylcholine (DMPE). Aggregation and fusion did not occur with single component (100 mol%) DMPC vesicles. Fusion was followed by two fundamentally different techniques, fluorescence resonance energy transfer which monitors intermixing of bilayers and ANTS-DPX which monitors intermixing of the sequestered aqueous interiors. It is suggested that a previously unreported role of ABA may be as a membrane fusagen. PMID- 2972287 TI - Site-directed mutagenesis at the regulatory site of fructose 6-phosphate-1-kinase from Bacillus stearothermophilus. AB - We have mutated Arg-25, Asp-59 and Arg-211 to alanine; and Asp-59 also to methionine, in fructose 6-phosphate-1-kinase from B. stearothermophilus (designated as RA25, DA59, RA211 and DM59 respectively). All four mutants did not change the affinity of the enzyme for ATP. RA25 has half the affinity for fructose 6-phosphate and exhibits sigmoidicity with respect to this substrate (Hill # = 2.0). DA59 has the same affinity for phosphoenolpyruvate (PEP) as the wild type whereas DM59 has 3-fold the affinity for this modulator and the inhibition is reversed by GDP. RA25 and RA211 are 100-fold less sensitive to PEP inhibition which is not relieved by GDP. It is concluded that Arg-25 and Arg-211, but not Asp-59, are involved in the direct binding of PEP and GDP. PMID- 2972288 TI - Purification of the D-2 dopamine receptor from bovine striatum. AB - The D-2 dopamine receptor has been purified 21500 fold from bovine striatal membranes. Solubilized receptor preparation was partially purified by affinity chromatography on a haloperidol adsorbent followed by gel filtration on a Sephacryl S-300 column. The fractions eluted from this column which contained the ligand binding activity were further chromatographed on wheat germ agglutinin conjugated to Sepharose. The resulting receptor preparation displays a major polypeptide band of an apparent molecular weight of 92 kDa, and exhibits a specific binding activity of 2490 pmol spiperone per mg protein. This purified receptor preparation can reabsorb specifically to the haloperidol affinity column indicating that the 92 kDa polypeptide represents the ligand binding unit of the D-2 dopamine receptor. PMID- 2972289 TI - Identification of a high-affinity receptor for interleukin-1 beta in rat brain. AB - A single type of high-affinity binding sites for IL-1 beta was identified in the rat hypothalamus (Kd = 1.0 +/- 0.2 nM) and cerebral cortex (Kd = 1.3 +/- 0.2 nM), but not in the pituitary. The maximum binding capacity (Bmax) in the hypothalamus (Bmax = 75.4 +/- 10.8 fmol/mg protein) was 4 times greater than in the cerebral cortex (Bmax = 17.2 +/- 1.5 fmol/mg protein). Neither various neuropeptides nor IL-2 appeared to influence the binding of [125I]IL-1 beta to the hypothalamic membrane preparations. The potency of unlabeled IL-1 alpha to replace the binding of [125I]IL-1 beta to the hypothalamic membrane preparations was considerably less than that of unlabeled IL-1 beta. These findings indicate that IL-1 beta receptors are heterogeneously distributed in the central nervous system and that IL-1 alpha does not bind with IL-1 beta receptors in the brain. PMID- 2972290 TI - Effects of the ortho-quinone and catechol of the antitumor drug VP-16-213 on the biological activity of single-stranded and double-stranded phi X174 DNA. AB - We have studied the effects of the recently reported two new metabolites of the antitumor agent VP-16-213, the ortho-dihydroxy derivative or catechol and the ortho-quinone, on the biological activity of single-stranded and double-stranded phi X174 DNA, the binding of the metabolites to calf thymus DNA and the conversion of the catechol into the ortho-quinone. Evidence was obtained for the oxidation of the catechol into the ortho-quinone and for the fact that the ortho quinone is the metabolite of VP-16-213 responsible for its binding to rat liver microsomal proteins. The catechol and ortho-quinone of VP-16-213 were found to bind 7-9 times more strongly to calf thymus DNA than VP-16-213 itself. In contrast to the parent compound VP-16-213, the catechol as well as the ortho quinone inactivated both single-stranded (ss) and double-stranded (RF) biologically active phi X174 DNA. The mean T37-values for inactivation of ss and RF phi X174 DNA by 2.2 x 10(-4)M catechol at 37 degrees and pH 7.4 were 96 and 640 min, respectively. Reduction of the ortho-quinone by NADPH cytochrome P-450 reductase resulted in formation of the catechol. The system ortho-quinone/NADPH cytochrome P-450 reductase inactivated ss phi X174 DNA with a mean T37-value of 454 min, and this inactivation was inhibited by DMSO. The mean T37-value for inactivation of ss phi X174 DNA by 1.8 x 10(-4) M ortho-quinone at 37 degrees and pH 4.0 was 24 min. The chemical stability of the ortho-quinone and the extent of inactivation of ss phi X174 DNA by the ortho-quinone were both pH-dependent: at higher pH the ortho-quinone was less stable and gave less inactivation of DNA. The aqueous decomposition product(s) of the ortho-quinone formed at pH 7.4 inactivated ss phi X174 DNA with a mean T37-value of 175 min. The rate of inactivation of RF phi X174 DNA by the ortho-quinone at pH 4.0 was twice as low as the rate of inactivation of ss phi X174 DNA: T37 = 49 min. When using excision repair deficient E. coli mutants (uvrA- or uvrC-), a higher inactivation of RF phi X174 DNA was found: T37 = 29 min for uvrA- E. coli, indicating that a part of the DNA damage introduced by the incubation with ortho-quinone is removed by excision repair.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2972291 TI - Antiinflammatory and antiallergic effects of a novel metabolite of Nocardiopsis sp. as a potent protein kinase C inhibitor from microbial origin. AB - The antiallergic and antiinflammatory effects of the new protein kinase C and calmodulin inhibitor K-252a (8R, 9S, 11S)-(-)-9-hydroxy-9-methoxycarbonyl-8 methyl-2,3,9,10-tetrahydro-8,11- epoxy-1H,8H,11H-2,7b,11a-triazadibenzo [a,g]cycl oocta [cde] trinden-1-one) were investigated in animal models, and the following results were obtained: 1. Oral administration of K-252a, ketotifen or oxatomide showed a dose-dependent inhibition on 48 h homologous passive cutaneous anaphylaxis in rats and anaphylactic bronchoconstriction in passively sensitized guinea pigs. 2. K-252a (1-100 mg/kg p.o.) exerted a dose-dependent protection against platelet-activating factor (PAF)-induced mortality in mice. This protective effect of K-252a was sustained for 7 h. 3. K-252a (1-100 mg/kg p.o.), as well as dexamethasone (1 mg/kg s.c.), showed remarkable inhibitory effects on rat paw edema induced by carrageenin, zymosan, PAF, and antigen-antibody reaction (passive Arthus reaction) and on the croton oil-induced rat ear edema. On the other hand, indomethacin (10 mg/kg p.o.) significantly inhibited carrageenin induced edema, but did not inhibit edema induced by other phlogistic agents. Based on these results it is suggested that K-252a, by oral administration, has antiallergic and antiinflammatory effects. PMID- 2972292 TI - Immunochemical characterization and quantitation of lipoprotein (a) in baboons. Development of an assay depending on two antigenically distinct proteins. AB - We have raised specific antibodies against the protein component of baboon lipoprotein (a) (Lp(a]. Apolipoprotein (apo) Lp(a) is a very large protein which separates into two distinct proteins, apo B and apo (a), when 2-mercaptoethanol is included during sample treatment for sodium dodecyl sulfate-electrophoresis. The antibodies were specific for baboon apo (a) and apo B. The presence of the two distinct antigens in the lipoprotein permitted the development of an enzyme linked immunosorbent assay that was specific for Lp(a) particles in serum. The assay could detect less than 1 ng of Lp(a) protein per well and was useful in the range of 1-9 ng. The assay was specific for Lp(a) and did not respond to other lipoproteins, such as low density lipoprotein. Lp(a) could be accurately quantitated in serum frozen at -80 degrees C in plastic tubing segments. Using the Lp(a) assay, the mean serum level of 80 unrelated baboons was 4.7 mg/dl, with the distribution skewed toward the lower levels. These data further support the value of the baboon as a model of the atherogenic lipoprotein Lp(a). PMID- 2972293 TI - Advances in electromyographic monitoring and biofeedback in the treatment of chronic cervical and low back pain. PMID- 2972294 TI - Clinical assessment of chronic low back pain. PMID- 2972295 TI - The child and technology. PMID- 2972296 TI - [Control of tuberculosis in Mexico]. PMID- 2972297 TI - [Sorbitol-gelatin and glutamic acid-lactose solutions as stabilizers of reference preparations of measles virus]. PMID- 2972298 TI - [In memoriam Clarence H. Moore, 1909-1988]. PMID- 2972299 TI - Marginal leakage of two glass ionomer cements: Ketac-Fil and Ketac-Silver. PMID- 2972300 TI - A case of congenital ectropion in Down's syndrome. AB - A rare case of primary congenital ectropion of all 4 eyelids in a child with Down's syndrome is reported to emphasise the problems of surgical management and to distinguish the condition from congenital eversion of the eyelids. Congenital ectropion is associated with other eyelid abnormalities and usually requires surgical measures to protect the cornea in contrast to congenital eversion which is characterised by the protrusion of oedematous conjunctiva from everted eyelids. This usually resolves spontaneously with simple supportive measures and no structural or functional eyelid abnormality remains. PMID- 2972301 TI - Antistaphylococcal activity of teicoplanin in Riyadh, Saudi Arabia. AB - The antistaphylococcal activity of teicoplanin, a new glycopeptide antibiotic, has been evaluated in vitro and compared with that of vancomycin, fucidin, rifampicin and gentamicin. Teicoplanin was found to be as active as vancomycin against clinical isolates of Staphylococcus aureus (both methicillin-resistant and methicillin-sensitive strains). Teicoplanin was found to be more active than vancomycin against Staphylococcus epidermidis. The minimum inhibitory concentration (MIC) of teicoplanin was 2- to 4-fold lower than that of other drugs tested. The minimum bactericidal concentration (MBC) of both teicoplanin and vancomycin either equalled or exceeded by 2-fold the respective MIC's. The rate at which staphylococci were killed showed that teicoplanin at 4 X MIC produced rapid killing in 10 h and no growth was detected after 24 h. PMID- 2972302 TI - Specialists in paediatric dentistry in the United Kingdom. PMID- 2972303 TI - Can social workers damage your dental health. PMID- 2972304 TI - Thrombin and plasmin activity in coronary artery disease. AB - Basal plasmin and thrombin activity in plasma were assessed by radioimmunoassay of the fibrinogen derivatives containing the sequence B beta 15-42 and of fibrinopeptide A respectively in a cross sectional controlled study of men with coronary artery disease. Compared with healthy controls (n = 33) men with angiographically defined coronary artery disease (n = 98) had a modest but significant increase in concentrations of fibrinopeptide A, indicating an activated coagulation system. Concentrations of B beta 15-42 were similar in those with coronary artery disease and in the controls. The enhanced thrombin activity in coronary artery disease is in keeping with current evidence suggesting an association between coronary artery disease and a hypercoagulable state. PMID- 2972305 TI - Measurement of evoked diaphragm twitch strength during anaesthesia. Adaptation and evaluation of an existing technique. AB - The technique of recording the mechanical response of the diaphragm after the transcutaneous stimulation of the phrenic nerve, during anaesthesia and surgery, is described in patients undergoing non-abdominal surgery. The pressure difference generated across the diaphragm after phrenic nerve stimulation was derived from the simultaneous recording of gastric and oesophageal pressures with air-filled balloon catheter systems. This was recorded as the diaphragm "twitch" in a manner similar to that of peripheral muscles. Special problems encountered in the use and the validation of the technique in the anaesthetized patient are considered. PMID- 2972306 TI - Effects of nadolol on arrhythmias during laparoscopy performed under general anaesthesia. AB - Cardiac arrhythmias are a well recognized complication of anaesthesia for laparoscopy. The effect of nadolol, given by mouth 12 h before operation, was compared with placebo on arrhythmias in 86 females undergoing laparoscopy. All types of arrhythmia were documented; there was a 97% incidence in the placebo group, but in the nadolol group there was a smaller incidence of supraventricular tachycardia, ventricular ectopics and atrioventricular dissociation (P less than 0.01). There was no significant difference in the incidence of sinus bradycardia. Nadolol may be recommended as a safe agent to be given by mouth before laparoscopy to reduce the frequency of cardiac arrhythmias during anaesthesia. PMID- 2972307 TI - Vascular responses to local atrial natriuretic peptide infusion in man. AB - 1. The effect on skin and muscle blood flow of arterial infusion of atrial natriuretic peptide (ANP) directly into the forearm circulation, and on venous tone of direct infusion into a dorsal hand vein, was studied in normal subjects. 2. ANP produced a dose-dependent increase in both skin and muscle blood flow, but at equivalent doses, produced no dilatation of noradrenaline-preconstricted dorsal hand veins. These findings indicate that ANP acting locally, is an arterioselective dilator in the upper limb circulation in normal man. 3. Measurements of ANP in venous plasma during arterial infusion suggest marked clearance of ANP across the forearm vascular bed. Such peripheral clearance may, at least in part, account for the short plasma half-life of this peptide. 4. The lowest dose of ANP infused was calculated to produce plasma levels similar to those found in patients with heart failure. The findings with this dose suggest that, in heart failure, circulating levels of ANP may be within a range capable of influencing peripheral vascular resistance directly. PMID- 2972308 TI - The influence of ipratropium bromide and sodium cromoglycate on benzalkonium chloride-induced bronchoconstriction in asthma. AB - 1. Benzalkonium chloride, an antibacterial preservative that is added to nebuliser solutions, has been shown to cause bronchoconstriction when inhaled by asthmatic subjects. 2. To investigate the potential role of reflex and mast cell dependent mechanisms in the pathogenesis of bronchoconstriction produced by benzalkonium chloride we examined the effects of ipratropium bromide and sodium cromoglycate on this response in both concentration-response and time-course studies in nine asthmatic subjects. 3. Pretreatment with inhaled ipratropium bromide (1 mg) and sodium cromoglycate (40 mg) displaced the benzalkonium chloride concentration-response curves to the right by a mean 2.2 fold and 3.1 fold respectively. 4. Ipratropium bromide and sodium cromoglycate markedly attenuated the airway response to benzalkonium chloride throughout the 45 min time course period, inhibiting the overall response by 56% and 78% respectively. 5. We conclude that benzalkonium chloride provokes bronchoconstriction in asthmatic subjects through a combination of mast cell activation and stimulation of peripheral and central neural pathways. PMID- 2972309 TI - The effect of oral retinoid therapy on the normal human immune system. AB - Twenty four patients were studied prior to and after 6 and 12 weeks therapy with isotretinoin (17 patients) for acne and related disorders, or with etretinate (7 patients) for psoriasis and related disorders. Patients treated with isotretinoin had a significant reduction in natural killer cell activity at an effector: target cell ratio of 100: 1 at 12 weeks and also a reduction in natural killer cell numbers at this time. Patients treated with etretinate had elevated natural killer cell activity and a significant elevation of natural killer cell numbers at 12 weeks. Other tests which were performed and showed no significant change at 6 or 12 weeks compared with starting levels included lymphocyte transformation in response to phytohaemagglutinin, pokeweed mitogen and concanavalin A, total numbers of circulating T lymphocytes, B lymphocytes and T helper and T suppressor subsets, numbers of epidermal Langerhans cells and serum levels of IgA, IgM and IgE. In view of the involvement of natural killer cells in the initial phase of organ rejection, these results suggest that isotretinoin is the safer of the two retinoids if administration to renal transplant recipients is considered, particularly in the immediate post-transplant period. PMID- 2972310 TI - Disseminated superficial porokeratosis and immunosuppression. AB - We present a patient who developed skin lesions typical of disseminated superficial porokeratosis (DSP) while on immunosuppressive therapy for pemphigus foliaceus. Phototesting with artificial light sources did not have any effect on the DSP lesions. The literature describing occurrence of DSP on immunosuppression is reviewed and possible pathogenetic mechanisms are discussed. PMID- 2972311 TI - Effect of isotretinoin on carbamazepine pharmacokinetics. PMID- 2972312 TI - Sequential dissociation of Ca2+ from the calcium adenosinetriphosphatase of sarcoplasmic reticulum and the calcium requirement for its phosphorylation by ATP. AB - The kinetics for dissociation of the stable enzyme-calcium complex of the sarcoplasmic reticulum calcium ATPase, cE.Ca2, were followed by assay with simultaneous addition of [32P]ATP and EGTA, which gives 70% phosphorylation of cE.Ca2 with k = 300 s-1 (25 degrees C, pH 7.0, 5 mM MgSO4, 0.1 M KCl). The binding of ATP to cE.Ca2 is described by kATP = 1.0 X 10(7) M-1 s-1, k-ATP = 120 s-1, and Kdiss = 12 microM; ATP binding is partially rate limiting for phosphorylation at less than 100 microM ATP. The sequential dissociation of Ca2+ from cE.Ca2 is described by k-2 = 55-60 s-1 for the first, "outer" Ca2+, k-1 = 25 30 s-1 for the second, "inner" Ca2+, and K0.5 = 3.4 microM, n = 1.9 (from Kdiss = 7.4 X 10(-7) M for Ca.EGTA). Dissociation of the inner Ca2+ is inhibited by external Ca2+, with K0.5 = k-1/k2 = 0.7 microM. This confirms the conclusion that dissociation of the two Ca2+ ions is sequential. The ability of cE.Ca2 to catalyze phosphorylation by ATP disappears in the presence of EGTA with k = 50-55 s-1, the same as k-2 for dissociation of the outer Ca2+ ion. This result, and the absence of the induction period that would occur if both cE.Ca2 and cE.Ca1 were catalytically competent, shows that both Ca2+ ions are required for phosphorylation. This conclusion is confirmed by the stoichiometry of 1.4/0.7 = 2.0 for the ratio of Ca2+ internalized to phosphoenzyme formed after simultaneous addition of ATP and EGTA. Phosphorylation of cE.Ca2 in the presence of 45Ca gives 0.15, not 0.3, 45Ca internalized, which corresponds to exchange of only 1 Ca2+ and is in agreement with this conclusion. The requirements for binding of two Ca2+ for catalytic specificity toward ATP and loss of two Ca2+ from E approximately P.Ca2 for specificity toward water account for the stoichiometry of Ca2+ transport and provide a possible reason for the two steps in the phosphorylation of cE.Ca2.ATP. PMID- 2972313 TI - Specificity of the sarcoplasmic reticulum calcium ATPase at the hydrolysis step. AB - The coupling of Ca2+ transport to ATP hydrolysis by the SR ATPase requires that the enzyme operate with considerable specificity, which is different at different steps. The limits of specificity of the calcium-free phosphorylated enzyme for transfer of its phosphoryl group to water have been examined. The rate of transfer of the phosphoryl group to the simple nucleophile methanol was compared to its transfer to water by following the formation of methyl phosphate from inorganic phosphate. The reverse reaction, hydrolysis of methyl phosphate, was compared to phosphate-water oxygen exchange. The reactions involving methanol as nucleophile or leaving group are at least 2-3 orders of magnitude slower than those involving water. This result indicates that the transition state for this reaction involves strong and specific interactions of the H2O molecule with the enzyme. These interactions may also involve the bound Mg2+ ion. The results also suggest that the difference in specificity between Ca2+ free and Ca2+ bound states of the enzyme involves significant differences in the structure of the catalytic site. PMID- 2972314 TI - Fluorescence resonance energy transfer within the complex formed by actin and myosin subfragment 1. Comparison between weakly and strongly attached states. AB - Fluorescence resonance energy transfer measurements have been made between Cys 374 on actin and Cys-177 on the alkali light chain of myosin subfragment 1 (S1) using several pairs of donor-acceptor chromophores. The labeled light chain was exchanged into subfragment 1 and the resulting fluorescently labeled subfragment 1 isolated by ion-exchange chromatography on SP-Trisacryl. The efficiency of energy transfer was measured by steady-state fluorescence in a strong binding complex of acto-S1 and found to represent a spatial separation between the two probes of 5.6-6.3 nm. The same measurements were then made with weak binding acto S1 complexes generated in two ways. First, actin was complexed with p phenylenedimaleimide-S1, a stable analogue of S1-adenosine 5'-triphosphate (ATP), obtained by cross-linking the SH1 and SH2 heavy-chain thiols of subfragment 1 [Greene, L. E., Chalovich, J. M., & Eisenberg, E. (1986) Biochemistry 25, 704 709]. Large increases in transfer efficiency indicated that the two probes had moved closer together by some 3 nm. Second, weak binding complexes were formed between subfragment 1 and actin in the presence of the regulatory proteins troponin and tropomyosin, the absence of calcium, and the presence of ATP [Chalovich, J. M., & Eisenberg, E. (1982) J. Biol. Chem. 257, 2432-2437]. The measured efficiency of energy transfer again indicated that the distance between the two labeled sites had moved closer by about 3 nm. These data support the idea that there is a considerable difference in the structure of the acto-S1 complex between the weakly and strongly bound states. PMID- 2972315 TI - Interaction of adriamycin with human erythrocyte membranes. Role of the negatively charged phospholipids. AB - The interaction of the antitumor compound adriamycin with human erythrocyte membranes, used as models of target cell membranes, has been studied using circular dichroism measurements. In order to elucidate the nature of the sites involved in the electrostatic interaction between adriamycin and erythrocyte membranes, its interaction with the following macromolecular systems was studied: phosphatidylserine-containing small unilamellar vesicles (SUV), prepared from total lipid extracts of erythrocytes, sialic acid-depleted erythrocyte ghosts and mucopolysaccharides. We have shown that the interaction between adriamycin and carboxylate groups is very weak and that negatively charged phosphate groups, in the case of membranes, or sulfate groups, in the case of mucopolysaccharides, are responsible for the prime interaction of adriamycin with these macromolecular systems. PMID- 2972316 TI - Regulation of calcium transport in drug-induced taurine-depleted hearts. AB - Drug-induced taurine depletion of rat heart led to the accumulation of free CoA, free carnitine and long-chain acylcarnitine, but a small decrease in long-chain fatty acyl-CoA. Although elevations in total tissue long-chain acylcarnitine levels have been linked to defective membrane function and the association of long-chain acylcarnitines with extramitochondrial membranes, these effects were absent in isolated sarcoplasmic reticulum prepared from taurine-depleted hearts. In contrast to the sarcoplasmic reticulum data, taurine depletion was associated with a significant decrease in ATP-dependent calcium uptake by isolated sarcolemmal vesicles. The major effect of taurine depletion on the sarcolemma was a 2-fold decrease in both the Vmax of calcium transport and the activity of the Ca2+ -stimulated ATPase. Sarcolemmal vesicles prepared from taurine-depleted hearts also exhibited a decreased capacity to transport calcium in exchange for sodium, although the initial rate of the process was unaffected by taurine depletion. Since incubation of sarcolemma from taurine-depleted hearts with taurine could not overcome the effects of taurine depletion, it was concluded that the effects of taurine were not caused by a direct interaction of it with the calcium pump. Possible mechanisms of taurine action are discussed. PMID- 2972317 TI - Ca2+ transport by rat liver plasma membranes: the transporter and the previously reported Ca2+-ATPase are different enzymes. AB - A rat liver plasma membrane fraction showed an ATP-dependent uptake of Ca2+ which was released by the ionophore A23187. This activity represents a plasma membrane component and is not due to microsomal contamination. The Ca2+ transport displayed several properties which were different from those of the high-affinity Ca2+-ATPase previously observed in these membranes (Lotersztajn et al. (1981) J. Biol. Chem. 256, 11209-11215; Birch-Machin, M.A. and Dawson, A.P. (1986) Biochim. Biophys. Acta 855, 277-285). These observations have shown that Ca2+-ATPase does not require added Mg2+ whereas we have demonstrated that, in the same membrane preparation, Ca2+ uptake required millimolar concentrations of added Mg2+. The Ca2+-ATPase has a broad specificity for the nucleotides ATP, GTP, UTP and ITP while Ca2+ uptake remains specific for ATP. Ca2+ uptake also displayed different affinities for free Ca2+ and MgATP compared to Ca2+-ATPase activity, with apparent Km values of 0.25 microM Ca2+, 0.15 mM MgATP and 1.0 microM Ca2+, 4 microM MgATP respectively. The apparent maximum rate of Ca2+ uptake was about 150 fold less than Ca2+-ATPase activity. These features suggest that the high affinity Ca2+-ATPase is not the enzymic expression of the ATP-dependent Ca2+ transport mechanism. PMID- 2972318 TI - The effect of calmodulin on the interaction of carbodiimides with the purified human erythrocyte (Ca2+ + Mg2+)-ATPase. AB - The activity of the solubilized and purified (Ca2+ + Mg2+)-ATPase from human erythrocyte membranes was inhibited by N,N'-dicyclohexylcarbodiimide in a concentration-dependent manner. The carbodiimide prevented formation of the phosphorylated intermediate during the catalytic cycle of the enzyme. Treatment of the enzyme with N,N'-dicyclohexyl[14C]carbodiimide resulted in the formation of a 14C-labelled polypeptide corresponding to the enzyme monomer (molecular weight 136,000). The tryptic fragmentation of this 14C-labelled enzyme resulted in the formation of three major 14C-labelled fragments with molecular weights of 58,000, 36,500 and 23,000, the latter two probably representing transmembrane and calmodulin-binding domains of the enzyme, respectively. In the absence of calmodulin, 6.7 molecules of N,N'-dicyclohexyl[14C]carbodiimide covalently bound to each molecule of Ca2+-ATPase; in the presence of calmodulin, the number of molecules of carbodiimide bound was 13.1. The binding of N,N' dicyclohexylcarbodiimide to the (Ca2+ + Mg2+)-ATPase greatly reduced its ability to bind to a calmodulin-agarose gel. PMID- 2972319 TI - The fms oncogene. PMID- 2972320 TI - The separation and characterization of two forms of Torpedo electric organ calelectrin. AB - Two methods for extracting calelectrin, a Ca2+-regulated membrane-binding protein from the electric organ of Torpedo marmorata, have been compared and the more promising one was modified to increase the yield to 7-8 mg.kg-1 wet weight of tissue, that is 4-5 times greater than the original method. The calelectrin so obtain could be resoloved into a minor component (designated L-calelectrin) eluted from an anion-exchange column at relatively low ionic strength (100 mM NaCl) and a major component (H-calelectrin) eluted at higher ionic strength (300 mM NaCl). The two forms were also separated by chromatography on a hydrophobic resin. Electrophoresis on cellulose acetate indicated that L-calelectrin had a lower mean isoelectric point that the H-form and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate showed that under reducing conditions (presence of 5% beta-mercaptoethanol) both forms migrated as single species, the L-form having a lower apparent relative molecular mass (Mr 32,000) that the H-form (34,000). Under non-reducing conditions, there was no change in the migration of L-calelectrin but the H-form was resolved into two components of Mr 34,000 and 32,000. The addition of 2 mM Ca2+ had no effect on the migration of either form. Both forms were equally recognized by an anti calelectrin antiserum and were microheterogeneous with respect to their isoelectric points (pH 4.3-5.5) in two-dimensional gel electrophoresis. Physical measurements were carried out on the major H-form. The Stokes radius was estimated to be 3nm, corresponding to an apparent Mr of 44,000. It was unaffected by changes in ionic strength, pH or Ca2+ concentration. Analytical ultracentrifugation gave a sedimentation constant of 2.9 S and an apparent Mr of 36,000. Measurements of circular dichroism indicated that 78% of the molecule was in the alpha-helix configuration and 22% in random coil. Ca2+ had no significant effect on the conformation. PMID- 2972321 TI - Ca2+- and phospholipid-binding properties of Torpedo electric organ calelectrin. AB - The Ca2+-regulated lipid-binding properties of the H- and L-forms of calelectrin present in the electric organ of Torpedo marmorata have been compared. Binding of H-calelectrin required Ca2+ in millimolar concentrations, whereas that of L calelectrin occurred in the micromolar range. Dissociation of H-calelectrin previously bound to lipids in the presence of 2 mM Ca2+ took place only when the Ca2+ concentration was reduced to micromolar concentrations. Binding was most effective to acidic phospholipids such as phosphatidylserine. Both forms of calelectrin promoted the aggregation of membrane vesicles in the presence of Ca2+.Mg2+, Na+ and K+ inhibited the Ca2+-induced binding to phospholipid, decreasing in effectiveness in that order. Binding was also inhibited by high pH. The surface activity and hydrophobicity index showed that H-calelectrin is a hydrophilic molecule. It may represent a less active, more highly phosphorylated "down-regulated" form of L-calelectrin. The role of calcium in H-calelectrin binding to lipid appeared to be consistent with the formation of a ternary complex of the protein, an acidic lipid and Ca2+, rather than with a direct interaction of lipid with hydrophobic sequences in H-calelectrin whose accessibility is Ca2+-regulated. PMID- 2972322 TI - End organ changes associated with the self-regulatory treatment of mild essential hypertension? AB - M-mode echocardiograms were obtained on unmedicated males with mild hypertension before and after treatment with thermal biofeedback, autogenic training, or self relaxation. Although patients for whom diastolic blood pressure (DBP) was successfully reduced showed trends toward reduction in left ventricular parameters while unsuccessful patients showed no changes, the results were not significant. For the four patients with borderline left ventricular hypertrophy, there was a strong trend (p = .06) for successful treatment to lead to a reduction in left ventricular mass. Moreover, across the whole sample, reduction in left ventricular mass was related (r = .30) to decrease in DBP. PMID- 2972324 TI - [Age-related differences in the inhibition of DNA synthesis in the liver of albino rats by proliferation inhibitors of chalone type]. AB - It was demonstrated that during postnatal ontogenesis, the heterogeneity of endogenous inhibitors of DNA synthesis in albino rat liver does not change. Upon ageing, the sensitivity of liver cells to the inhibiting effect of chalone containing extracts and the functional activity of the inhibitor are diminished. PMID- 2972323 TI - [ATP-dependent transport of Ca2+ in myocardium sarcolemma vesicles and its activation by phorbol esters]. AB - Highly purified pig myocardium sarcolemma vesicles possess the Ca2+,Mg2+-ATPase activity (4.1 mumol Pi/mg protein/hour) and induce the ATP-dependent accumulation of 45Ca2+ (6.0 nmol/mg protein/min). This reaction is not stimulated by oxalate; Ca2+ are released from the vesicles by saponin and Na+ treatment, which suggests that Ca2+ transport against the concentration gradient is induced by myocardium sarcolemma vesicles and not by sarcoplasmic reticulum fragments. The phorbol ester possessing a biological activity of a growth-promoting factor and activating membrane-bound protein kinase C stimulates the Ca2+,Mg2+-ATPase activity and the ATP-dependent accumulation of Ca2+, whereas its counterpart devoid of biological activity does not influence Ca2+ transport. Polymixin B, a specific inhibitor of protein kinase C, prevents the activating effect of phorbol esters on Ca2+ accumulation inside the vesicles. It is suggested that the ATP dependent transport of Ca2+ in myocardium sarcolemma is controlled by Ca2+ phospholipid-dependent phosphorylation catalyzed by protein kinase C. PMID- 2972325 TI - Immunocytochemical localization of heparan sulfate proteoglycan in human decidual cell secretory bodies and placental fibrinoid. AB - A distinct ultrastructural feature of human decidual cells is the presence of membrane-bound secretory bodies, 0.3-0.5 micron in diameter, located within club shaped processes at the cell periphery. These secretory bodies contain 30-60 nm electron-dense granules. Using specific antibody and the protein A-gold technique, we examined the localization of heparan sulfate proteoglycan in human decidual cells. Morphometric analysis of gold particles in cellular compartments was performed with a Zeiss Videoplan computer system. Immuno-gold staining was present in the decidual cell cytoplasm and the extracellular space, especially in the zone of the external lamina. Gold particles, indicating the locale of heparan sulfate proteoglycan, were concentrated over the electron-dense granular material within decidual secretory bodies contained in club-shaped processes at the cell periphery. Immunolabeling of placental fibrinoid was also observed. This report provides the first identification of a specific molecular constituent of decidual secretory bodies and indicates a role for these structures in secretion of the peri-decidual cell extracellular matrix. PMID- 2972326 TI - In vitro inhibition of normal human hematopoiesis by marrow CD3+, CD8+, HLA-DR+, HNK1+ lymphocytes. AB - We previously demonstrated that after allogeneic bone marrow transplantation (BMT) a subset of CD8, HNK1, and DR-positive T lymphocytes are able to inhibit CFU-GM and BFU-E growth with an HLA-DR restriction. In this study we investigated whether these cells, present in normal marrow in low concentration (less than 1%), play the same role. HNK1-positive sorted marrow cells forming rosettes (E+C) were able to inhibit BFU-E and CFU-GM growth when added back to the marrow E-C at a ratio of 1:10 (HNK1+ E+C/E-C) in a range from 40% to 60%. This inhibitory effect was also detected for a cellular ratio of 1:100, which is the normal marrow value for this subset of T cell. HNK1+ DR+-sorted E+C after double immunofluorescent labeling also showed the same inhibitory activity as the HNK1+ E+C, whereas the negative fraction including all the other E+C had no detectable inhibitory activity. CD3 and CD8 antigens were also present on the membrane of these cells, as demonstrated in two cases by double-immunofluorescent labeling performed with anti-CD3 or anti-CD8 monoclonal antibodies (MoAbs) and HNK1 MoAb, respectively, and subsequent cell sorting. Blocking experiments, performed by adding in culture anti-CD4 and anti-CD8 MoAbs to HNK1+ T cells showed that only the last MoAb was able to prevent inhibition of hematopoietic colony growth. These results confirmed that one subset of CD3+, CD8+, HNK1+, and DR+ T cells was responsible for in vitro inhibition of normal hematopoiesis. In addition, this inhibition was genetically restricted to HLA-class II antigens, since in three co culture experiments with unrelated bone marrow cells inhibition occurred only when cells with one haplo-identical HLA-DR antigen was added back to the culture. Indeed, this effect was really HLA-DR restricted, since in blocking experiments with different anti-HLA class II MoAbs (anti-DR, anti-DP, and anti-DQ MoAbs) only an anti-HLA-DR MoAb was able to prevent the colony growth inhibition by CD3+ HNK1+, or CD8+ HNK1+ E+C. In conclusion, the CD3+, HNK1+, CD8+, DR+ cells may be the T-cell subset able to inhibit normal hematopoiesis with an HLA-DR restriction. PMID- 2972327 TI - Natural killer activity is not dependent on the CD3-Ti T-cell receptor antigen complex. PMID- 2972328 TI - Decreased Ca pump ATPase activity associated with increased density in human red blood cells. AB - Red blood cells (rbcs) from five different normal humans were separated according to density using a simple procedure. The procedure involved centrifugation for 30 minutes in small glass tubes in the absence of any density gradient medium. This produced a column of rbcs arranged according to their density. Samples of the top 8% of the columns and bottom 8% of the columns were removed from the tubes with a micropipet. From each donor, samples of the least and most dense cells, respectively, were pooled from multiple tubes for each donor and designated "top" and "bottom" cells. These top and bottom cells were compared with unselected (total) cells from the same subjects, respectively. Top cells were larger and bottom cells were smaller than total cells. ATPase activities were operationally defined and measured in saponin lysates of these rbcs. The Ca pump ATPase (both in the calmodulin-activated and calmodulin-independent states [achieved by addition of compound 48/80]) of the top cells exhibited greater activity, and the Ca pump ATPase of bottom cells exhibited lower activity than total cells. It was suggested that loss of Ca pump ATPase activity is associated with rbc aging and may be a determinant of rbc life span. A mechanism for the loss of Ca pump ATPase activity was suggested. This speculative mechanism is based upon selective proteolysis of the Ca pump ATPase by the Ca-activated protease, calpain. PMID- 2972329 TI - Ectopic pregnancy. PMID- 2972330 TI - Molecular genetics in clinical practice: evolution of a DNA diagnostic service. AB - The development of a molecular genetics diagnostic service over a three year period was studied in a National Health Service region with a population of three million. Starting from a time when few diagnostic applications were possible, the number of disorders and the overall demand had grown rapidly. Conditions for which molecular genetic diagnosis had been provided included Duchenne and Becker muscular dystrophy, myotonic dystrophy, Huntington's disease, and cystic fibrosis. Of 405 requests for diagnosis, 151 (37%) related to determination of carrier state, 187 (46%) to determining the feasibility of future prenatal diagnosis, and 67 (17%) were prenatal diagnostic biopsy samples, almost exclusively of first trimester chorion. DNA samples for future diagnostic use with a wide range of genetic disorders had also been banked. The study showed a need for close integration with clinical genetics services to allow satisfactory genetic counselling and interpretation of risks. PMID- 2972331 TI - Screening for Down's syndrome. PMID- 2972332 TI - Nickel chloride protection against alloxan- and streptozotocin-induced diabetes. AB - The ability of NiCl2 to prevent alloxan- and streptozotocin-induced hyperglycemia was tested in rats. NiCl2 injected before alloxan and streptozotocin prevented the hyperglycemic response to the drugs. The protective effect of NiCl2 was linked to an increase in the specific activity of Cu-Zn superoxide dismutase (SOD). The effect of NiCl2 on SOD activity might be related to the effect of nickel on calcium, copper and zinc concentrations. PMID- 2972333 TI - Construction and reconstruction of memories. AB - 1. Recent evidence suggests that treatments given after training may influence memory in two ways: by becoming themselves incorporated to the experience, or by altering post-training mechanisms involved in the storage of the experience. The two processes may be called consolidation. 2. Some endogenous substances that are normally released during or after training (brain beta-endorphin; the peripheral stress hormones, ACTH, epinephrine and vasopressin) appear to be of particular importance. Their effect may become incorporated to the experiences as a conditioned stimulus (CS), generating state dependency. The effect of beta endorphin appears to be physiological, since the substance is released by novel experiences. 3. Post-event information provided by other training experiences, in rats, or by comments or leading words, in humans, may also incorporate to the experiences, altering their content qualitatively or quantitatively. 4. A variety of substances including the stress hormones at low doses and analeptic drugs may facilitate retention when given after training. In this case, the effect is best explained by an enhancement of the post-training strengthening of memory traces. 5. The reiteration of part of the experiences at the time of testing facilitates retrieval. This may be viewed as a reconstruction of consolidation at the time of retrieval, and may be obtained using cognitive material ("priming"), or neurohumoral stimuli (a beta-endorphin injection, or a presumable release of brain beta-endorphin by an interpolated novel experience). The effect can be seen in animals rendered amnestic by electroconvulsive shock, and in humans with amnesia of organic and non-organic nature. 6. The human amnesic syndrome seems, thus, largely explainable by a deficit of retrieval. It is possible that the stimulation of retrieval by priming, or by drugs, through the "reconstruction" of consolidation, may be useful for the relief or treatment of the human amnesic syndrome. PMID- 2972334 TI - Breast reconstruction by the free lower transverse rectus abdominis musculocutaneous flap. AB - Free TRAM flap transfer for breast reconstruction following mastectomy overcomes the shortcomings of the pedicled TRAM flap. It ensures the perfusion of the entire flap via its dominant vascular pedicle and allows for flexibility in the design of the ensuing breast mound. The authors' experience with ten free lower TRAM flap transfers is reviewed. The present surgical technique is described and three cases are presented to illustrate its application. The advantages and limitations of the method are compared particularly with its pedicled version. PMID- 2972335 TI - Rectus nerve entrapment causing abdominal pain. PMID- 2972336 TI - Responses of cat muscle spindle afferents after short periods of denervation and reinnervation. AB - The functional recovery of cat muscle spindles has been examined following short periods of denervation and reinnervation. The restoration of function was more rapid than after longer periods of denervation though the firing rates showed a similar reduction except after the shortest recovery periods when a number of highly dynamic afferents were recorded. PMID- 2972337 TI - Microinjection of carbachol into the pontine area is unable to modify insomnia induced by p-cholorophenylalanine (PCPA). AB - It has previously been shown that cholinergic drugs administered into the pontine area (field tegmental gigantocellular, FTG) are capable of inducing significant increases of rapid eye movement (REM) sleep periods in normal cats. The purpose of this study was to determine whether such cholinergic stimulation is capable of inducing similar effects in p-chlorophenylalanine (PCPA) insomniac cats. Four groups (n = 6) of cats were used in this study. The first two groups received a microinjection of either, saline (1 microliter) or carbachol (8 micrograms/1 microliter). The last two groups, prior to the microinjection, received PCPA (400 mg/kg) pre-treatment. Histological analysis showed that the tips of the cannulae were located in the caudal FTG, at the level of the ponto-medullary union. Results showed that carbachol significantly increased REM sleep in normal cats as compared to saline, whereas, carbachol was unable to modify the insomnia induced by PCPA. These findings suggest that cholinergic effects on REM sleep require a serotoninergic system. The results are discussed in terms of acetylcholine/serotonin interaction in the control of REM sleep. PMID- 2972338 TI - [Radiosensitivity of chromosomes and the kinetics of division of peripheral lymphocytes in individuals with an aneuploid karyotype]. PMID- 2972339 TI - Inhibition of thymidylate synthase after administration of doxifluridine in a transplantable colon carcinoma in the rat. AB - Parameters for inhibition of thymidylate synthase (TS) in a DMH-induced transplantable rat colon carcinoma were studied after intraperitoneal administration of bolus doxifluridine (5'-dFUR) 200 mg/kg. Levels of 5'-dFUR, 5 fluorouracil (5-FU), fluorouridinediphosphate (FUDP), and fluorouridinetriphosphate (FUTP) were determined by use of high-performance liquid chromatography. Micromethods for analysis of 5-fluoro-2'-deoxyuridylate (FdUMP) and TS were used to study the in vivo intracellular pharmacokinetics of TS inhibition. Peak values of 5'-dFUR and 5-FU were found at 30 min and showed exponential declines with values close to zero at 5 hr. Substantial levels of FUDP and FUTP were found throughout the 24 h observation time. Peak FdUMP levels were modest compared to those observed after equimolar administration of 5-FU, but FdUMP persisted in amounts well above available binding sites on TS for the 24 h observation time. Reduction of free TS enzyme to undetectable levels (less than 0.05 pmol/g) lasted for 4 h, and at 24 h, there was still almost 70% enzyme inhibition. The total amount of TS (TStot) defined as free [3H]FdUMP-titrable enzyme (TSf) plus TS bound to FdUMP in a ternary complex (TSb) increased as a result of 5'-dFUR bolus injection from 15 to 50 pmol/g during the 24 hr observation time. We conclude from these data that 5'-dFUR is converted to 5-FU and subsequently to FdUMP, and the results suggest that 5'-dFUR exerts its cytotoxic effects through inhibition of TS and incorporation into RNA. PMID- 2972340 TI - [Ingestion of soybean epoxide oil. Effects on monooxygenases, epoxide hydrolase and activities of UDP glucuronosyltransferases in hepatic microsomes of the rat]. AB - The effect of peroxidized soybean oil in the diet of male Wistar rats was studied on hepatic drug metabolizing enzymes and their phenobarbital induction and compared to that of natural soybean diet in the same conditions. No hepatomegaly or increase in serum transaminases occurred, however growth was inhibited after ingestion of peroxidized soybean oil. In addition, the protein biosynthesis of epoxide hydrase determined by immunochemistry was largely stimulated by this treatment; but the corresponding activity measured with benzo(a)pyrene 4-5 oxide as a substrate was increased in weaker proportions. This induction was limited to epoxide hydrolase only, since the enzymes of phase one were not affected and UDP glucuronosyltransferase activities toward group I substrates were randomly activated. The induction of epoxide hydrolase may affect only one or several isoforms of the membrane enzyme which are not necessarily specific to benzo(a)pyrene 4-5 oxide activity determination of the enzyme. PMID- 2972341 TI - [Immunocytologic study of light cell lines established in vitro from Ewing's sarcoma. Identification of neural markers]. AB - Using immunocytological techniques, neuroectodermal markers were identified on Ewing's sarcoma cell lines established in vitro and carrying the chromosomal translocation t(11;22). Eight cell lines were tested using a panel of monoclonal antibodies. The presence of cell surface antigens recognized by HNK-1 antibody was confirmed. The cells showed also positive reactions using antibodies directed against Neuron-Specific-Enolase and neurofilament proteins. The presence of these neural markers in the Ewing's sarcoma cells tested is an additional argument substantiating the putative neural origin of this tumor. PMID- 2972342 TI - [Preliminary study on nerve regeneration through an autologous venous tube]. AB - Large posttraumatic defects in the peripheral nervous system bring up serious problems to the surgeon. There is no definitive answer yet. The authors present a model for nervous regeneration studies; venous autograft was used to bridge defects in a divided rat sciatic nerve; the venous tube guided the regenerating axons towards the distal stump over up to 1.9 cm length. Preliminary studies about nervous regeneration stimulation with autologous Schwann cells are discussed. PMID- 2972343 TI - [Peripheral distribution of a neuropeptide recognized by an antiserum raised against the mammalian CRF41, in the mollusc Helix pomatia]. AB - Immunocytochemical methods using an antiserum raised against ovine corticoliberin revealed perikarya and processes in the central and peripheral nervous system of the Pulmonate Gastropod Helix pomatia. The coexistence of immunoreactive nerve fibres and primary sensory neurons in the intestinal wall and in the tentacles appeared particularly significant. The distribution of this peptide suggests that it could act as a sensory neurotransmitter at the central and peripheral levels. PMID- 2972344 TI - [Cross-reactivity between parasitic antigens and T lymphocyte surface antigens: a possible host-escape mechanism for Echinococcus multilocularis]. AB - We report an immunocytochemical analysis of E. m. protoscolices obtained in 2 strains of mice (AKR, Balb c) which were experimentally infected. Sections of hepatic and peritoneal lesions and spreading of protoscolices from peritoneal vesicles were analyzed. Five monoclonal antibodies, specific of murine T lymphocyte populations, produced an intense and regular staining of the anterior area of the protoscolices. This immunostaining has not yet been explained; it could bear witness to particular mode of parasite protection against host immunological responses. PMID- 2972345 TI - [Preservation of the functional integrity of cellular material after cell microencapsulation by polymerization]. AB - A new process for microencapsulating living cells is described. A cross linked network was polymerized using acrylamide and bisacrylamide monomers round agarose embedded cells in an hydrophobic medium. The reaction was initiated by excitation of Vit. B2 under visible irradiation. The absence of toxic effect was evaluated from the insulin secretion of encapsulated islets of Langerhans. PMID- 2972346 TI - [Evidence for several storage and exportation pathways of pancreatic digestive enzymes during protein malnutrition followed by refeeding]. AB - Outputs of pancreatic juice hydrolase were constant during a three-hour collection in control rats fed 20% protein and in malnourished rats fed on a low protein diet (5% protein). However, during the same time after a 3H-Leucine pulse, the relative proportion of each labelled enzyme (newly synthesized) were changed. In control rats, this phenomenon was specially obvious for trypsinogen 2 (Tg2) and chymotrypsinogen 1 (Chtg1), and support the existence of separate pools for various pancreatic hydrolases. In malnourished rats, the relative proportion of labelled Tg2 is not modified by protein malnutrition, whereas the proportion of labelled amylase, lipase and Chtg1 were decreased. Refeeding carried out the same labelling profile of each enzyme in the two groups. PMID- 2972347 TI - [Nutritional utilization of cottonseed flour, compared to soya flour, by growing rats with protein malnutrition]. AB - The aim of this study was to compare the nutritional value of three diets with rats submitted to protein malnutrition. After one month on a low protein diet (2% casein), the rats were refed with corn-soya-milk (60%/30%/5%, W/W/W) and rice cotton-milk (60%/30%/5%) diets. This produced a higher growth rate than either the corn-cotton milk (60%/30%/5%) diets or a control diet containing 20% casein. The caloric intake/100 g body weight was about the same with the four rehabilitation diets. The highest apparent nitrogen digestive utilization coefficients (C. U. D.) values were obtained with the control diet, lower values with the rice-cotton-milk diet, then the corn-cotton-milk diet and lowest with the corn-soya-milk diet. Results of lipid C. U. D. were in the same order. The highest nitrogen balances were obtained with the control diet, then the corn-soya milk diet, the rice-cotton-milk diet and lowest values with the corn-cotton-milk diet. Our data prove that the cotton flour mixed with rice may perfectly replace soya flour associated with corn in a nutritional rehabilitation diet. PMID- 2972348 TI - Atrial natriuretic factor in the pediatric intensive care unit. AB - ANF is a newly discovered peptide hormone that has significant implications for critical care physicians. This hormone, released from the heart, is especially responsive to fluid challenges as well as to many of the drugs commonly used in the ICU, including pressor and anesthetic agents. It has potent arterial vasodilating effects in pharmacologic doses and may be an important natural vasodilating agent, especially in the renal vascular bed. In patients on dopamine, it may potentiate the renal vasodilating effect and may provide an effective therapy for developing acute renal failure. Children with congenital heart disease and patients with CHF have elevated levels that clearly alter the aldosterone-angiotensin II system and may help us to understand and treat these conditions more effectively. Additionally, ANF may be a marker for adequacy of treatment in these disease states. The potential uses for ANF include diuresis in patients with fluid overload and diuretic resistance, treatment of CHF, and as a short-acting vasodilator. In the ICU, many therapies affect cardiac pressures and volume regulation. Positive-pressure ventilation may decrease the release of ANF by decreasing venous return and thus contribute to water retention. Drugs used in the ICU may directly affect ANF levels and markedly affect the homeostasis of fluid and electrolyte balance. This hormone system interacts intimately with renin, angiotensin II, and aldosterone. These interactions may play a significant role in the development of essential hypertension. Although not addressed in this article, the treatment and understanding of essential hypertension may be significantly advanced by understanding these relationships. It is clear that ANF acts as a hormone with complex interactions between the heart, volume status, electrolyte balance, renin-angiotensin II-aldosterone, vasopressin, and vascular tone. Although currently no definitive picture exists for these complex interactions, this is an exciting new hormone with significant implications for patient management in the ICU. As research continues, the picture will become clearer and our understanding of this new hormone more precise. PMID- 2972349 TI - Patient communication tools in a pediatric intensive care unit. PMID- 2972350 TI - Effects of different thyroid treatments on the biochemical characteristics of rabbit myocardium. AB - It is well established that extreme dysthyroidism drastically alters the biochemical character of cardiac muscle. The purpose of this study was to determine if minor thyroid treatments would result in significant changes in the character of three major biochemical systems of muscle: metabolic; calcium regulating; and contractile systems. Different groups of New Zealand white rabbits had continuous time release propylthiouracil (PTU) pellets (500, 300, 200 and 100 mg) or triiodothyronine (T3) pellets (15 and 25 mg) subcutaneously implanted for 21 days. The ventricular myosin phenotypes shifted from 92% V3 myosin in the control rabbit hearts to 55% V3 and 10% V3 in the 15 mg and 25 mg T3 groups, respectively. PTU treatment resulted in a complete shift to the V3 myosin isoform. The sarcoplasmic reticulum Ca2+-ATPase activity increased with T3 and decreased with PTU treatments, except in the 500 mg PTU group. Ca2+-ATPase activity in the groups either side of the euthyroid group (100 mg PTU and 15 mg T3) did not change significantly. The glycolytic or aerobic potentials of the myocardium did not change with any of these minor thyroid treatments. It was concluded that the metabolic, enzymes, sarcoplasmic reticulum Ca2+-ATPase and myosin isozymes have different sensitivities to thyroid treatment and that minor thyroid treatments do result in significant changes in the biochemical character of the myocardium. These findings indicate that subclinical deviations in euthyroid status may affect myocardial biochemical character. PMID- 2972352 TI - Hepatitis: the law and the operating room nurse. PMID- 2972351 TI - Characterization and metabolism of canine proximal tubules, thick ascending limbs, and collecting ducts in suspension. AB - Preparations of distinct nephron segments were obtained from dog kidneys by collagenase treatment. Four morphologically different tissues were isolated: glomeruli, proximal tubules, thick ascending limbs, and papillary collecting ducts. Each segment possessed a characteristic assay of membrane-bound and cytoplasmic enzymes. Specific metabolic characteristics also were found: gluconeogenesis and ammoniagenesis in proximal tubules, glycolytic aerobic metabolism in thick ascending limbs, and glycolytic anaerobic metabolism in papillary collecting ducts. The assay of Na+ -K+ ATPase, H+ -ATPase, and Ca2+ ATPase activities in these nephron segments demonstrated a specific enrichment of Na+ -K+ ATPase in thick ascending limbs, and of H+ -ATPase in proximal tubules and papillary collecting ducts. Tubular respiration in the absence or presence of ouabain, 1,3-dicyclohexylcarbodiimide, or furosemide demonstrated that the respiration of each segment could be correlated to the activity of specific ion motive ATPases. Furthermore, a tight coupling between ion transport, ATP turnover, and substrate oxidation was demonstrated. These isolated tubular structures are thus viable and capable of transepithelial transport. Our preparation provides large amounts of defined population of tubules and are thus useful for the study of biochemical and functional heterogeneity along the nephron. PMID- 2972353 TI - Perceived benefits of a lifestyles modification program in the rehabilitation of Worker's Compensation recipients. PMID- 2972354 TI - Combined floxuridine and cisplatin in a fourteen day infusion. Phase I study. AB - Twenty patients received 28 courses of 5FUDR (floxuridine) admixed with Cisplatin (CDDP) and administered as a continuous infusion for 24 hours for 14 consecutive days. Pharmaceutical studies of the admixture of 5FU with CDDP and 5FUDR with CDDP demonstrated that only 5FUDR was compatible with CDDP and that the admixture was stable for 7 days. This Phase I study established the optimal dose rate for the individual components of the admixture and demonstrated that CDDP decreases the maximum tolerated dose rate for 5FUDR. The optimal dose rate for 5FUDR is 0.075 mg/Kg/d, and for CDDP the optimal dose rate is 7.5 mg/M2/d. Dose rate limiting toxicity is an enteritis which is radiographically similar to regional enteritis and is related to the 5FUDR. An ancillary finding was a significant decrease in serum magnesium levels in 11 of 13 monitored courses presumably related to the platinum. PMID- 2972355 TI - Ultrastructural localization of sulphate glycoconjugates in the human glomerular capillary wall using the high iron diamine method. AB - The distribution of fixed anionic sites within glomerular capillary walls has been studied in man by applying two ultrastructural histochemical methods--the high iron diamine and dialysed colloidal iron methods--to tissue chopper sections and to isolated glomeruli obtained from surgical fragments of renal tissue. By using the high iron diamine method we have been able to demonstrate that in man, too, there are sulphate (possibly heparan sulphate proteoglycan) sites preferentially located in the lamina rara esterna of the basement membrane and in the cell coat of the urinary surface of podocytes. Non-sulphate (high iron diamine-negative, dialysed colloidal iron-positive) anionic sites have been identified not only in the glycocalyx of the epithelial and endothelial cells but also in the laminae rarae of the basement membranes, where they show a more extensive distribution pattern than sulphate sites. The proposed methods seem particularly suitable for the study of human renal tissue; they could, in fact, provide useful information about the behaviour of the various anionic components of the glomerular capillary wall in pathological conditions. PMID- 2972356 TI - Cytogenetic findings in leukemic cells of 56 patients with constitutional chromosome abnormalities. A cooperative study. Groupe Francais de Cytogenetique Hematologique. AB - Fifty-six patients with a major constitutional chromosome anomaly and a malignant hematologic disorder in whom the chromosomes of the malignant cells were analyzed by banding techniques are reported from a cooperative and retrospective study. The patients included 25 with trisomy 21, nine with a sex chromosome anomaly, and 22 with a structural rearrangement. For Down's syndrome patients, the morphologic and cytogenetic aspects of acute leukemia were studied because it has been postulated that there could be a specific pattern of acute leukemia in Down's syndrome patients. For other chromosome rearrangements detailed karyotypic changes are reported, but the question of whether these patients are at greater risk of hematologic malignancy remains unanswered. PMID- 2972357 TI - Infiltration of interleukin-2-inducible killer cells in ascitic fluid and pleural effusions of advanced cancer patients. AB - Using ascitic fluid or pleural effusion obtained from 13 ovarian or metastatic breast cancer patients, we separated tumor cells from effusion-associated lymphocytes (EAL) with Percoll density centrifugation. Lymphocytes were incubated with recombinant interleukin 2 (IL-2) for 3-4 days and then assessed for tumoricidal activity in a 51chromium-release assay. The IL-2-activated EAL were found to lyse autologous fresh tumor cells, as well as allogeneic fresh tumor cells and FMEX tumor cells, a melanoma cell line which is resistant to natural killer cell activity but is sensitive to lysis by lymphokine-activated killer cells. There was little or no tumoricidal activity seen in freshly isolated EAL or in EAL which were cultured in medium without IL-2. Phenotypically, the IL-2 activated EAL were largely CD3-, although some cytolytic activity was found in CD3+ populations. Also, most activity was found in cells positive for CD2 (OKT11) and CD16 (Leu 11b), and negative for the monocyte marker Leu M3. These results indicate that the activated cell types found in EAL were predominantly natural killer/lymphokine-activated killer-like with a small contribution from T-cells. Finally, EAL were readily activated by IL-2 in medium containing autologous effusion fluid, indicating that in situ activation of tumoricidal activity by IL 2 can occur in the face of potentially inhibitory substances or cells that may exist in the effusions. Direct introduction of IL-2 may therefore be a potential therapeutic modality of effusion-forming cancers. PMID- 2972358 TI - Insulin-like growth factor 1 receptors in human breast cancer and their relation to estradiol and progesterone receptors. AB - Insulin-like growth factor 1 (IGF1) binding sites were characterized in breast cancer. We demonstrate the presence of one high affinity binding site. Chemical cross-linking of 125I-IGF1 to breast cancer membranes in reducing condition and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed one band with an apparent molecular weight of 130,000. The specificity of the binding was studied. IGF2 was a good competitor whereas insulin competed with a potency lower than 1/100 that of IGF1. This IGF1 binding corresponded to the previously described type 1 IGF receptor (IGF1-R). IGF1-R was determined in 76 human breast cancer biopsies. Ninety-three % of the tumors were positive. The specific binding range was 0-16.4%; the geometric IGF1-R mean level was 3.9%. There was a relation (chi 2 test) between IGF1-R and progesterone receptor positivity rates (P = 0.002). The IGF1-R concentrations were correlated (Spearman test) with those of estradiol receptor (P = 0.0018) and progesterone receptor (P = 0.0011). A positive linear correlation existed between IGF1-R and estradiol receptor (P = 0.006) and between IGF1-R and progesterone receptor (P = 0.003). Our demonstration of the presence of IGF1-R in human breast cancer biopsies suggests that IGF1, acting either via the endocrine, paracrine, or autocrine pathways, could stimulate tumor growth. PMID- 2972359 TI - Regulation of tumor-induced myelopoiesis and the associated immune suppressor cells in mice bearing metastatic Lewis lung carcinoma by prostaglandin E2. AB - The in vitro and in vivo effects of prostaglandin E2 (PGE2) and of its stable analogue, 16,16-dimethyl-PGE2 (dmPGE2), on myelopoiesis and on the myelopoiesis associated immune suppressor cell activity of mice bearing metastatic variant Lewis lung carcinoma (LLC-C3) tumors are assessed. In vitro studies showed a reduced susceptibility of bone marrow myeloid progenitor cells (CFC) from LLC-C3 tumor bearers versus normal mice to the growth-inhibitory effects of PGE2. When added to cocultures of bone marrow cells with LLC-C3 supernatants, PGE2 lessened the frequency of CFC and slightly reduced the generation of bone marrow immune suppressor cells. In vivo studies showed that 4 daily injections of dmPGE2 into LLC-C3 tumor-bearing mice caused some reduction in femoral bone marrow CFC and had an insignificant effect on bone marrow suppressor cell activity. In contrast to the relative insensitivity of bone marrow cells of tumor bearers to the effects of PGE2, in vitro studies showed that CFC formation by spleen cells of tumor bearers was readily inhibited by PGE2. Likewise, in vivo studies showed that spleen cells of dmPGE2-treated LLC-C3-bearing mice had a reduction in cellularity, CFC, and the level of spontaneous proliferation; a reduction in suppressor cell activity; and an increase in blastogenesis. Thus, short-term dmPGE2 treatment of LLC-C3-bearing mice limited the tumor-induced splenic myelopoiesis and reduced the associated splenic immune suppressor cell activity. PMID- 2972360 TI - Copolymer-I therapy for multiple sclerosis. PMID- 2972361 TI - Immunotherapy of advanced gynecologic cancer patients utilizing mumps virus. AB - Immunotherapy utilizing mumps virus (MV) was applied to 22 patients with advanced gynecologic cancer. Therapy schedule was based on the established concept of augmenting antitumor immunity utilizing virus-reactive helper T cell (Th) activity in a mouse system. Briefly, presensitization of cancer patients was performed subcutaneously (s.c.) with 10(8) PFU of mumps virus (MV). After increasing the anti-MV antibody activity of patients, 10(9) PFU of MV was administered locally or systemically. Marked clinical response was obtained by local (intraperitoneal or intrathoracic) injection of MV in patients with ascites or pleural fluid who had received preimmunization with MV. However, unprimed patients or patients with large tumor mass did not respond well to MV therapy. None of the patients showed untoward response to MV therapy except for controllable transient fever. PMID- 2972362 TI - Enhancement of murine mixed lymphocyte response by 1,1-dimethylhydrazine: characterization and possible mechanism. AB - Treatment of mice with 1,1-dimethylhydrazine (UDMH) resulted in enhancement of the one-way mixed lymphocyte response (MLR); this effect was seen when both responder and stimulator mice were treated as well as when just the stimulator or just the responder mice were treated. Experiments in which splenocytes were exposed to UDMH in vitro indicated that exposure of the stimulator cells alone resulted in an enhanced MLR; exposure of the responder cells alone had no effect; and addition of UDMH to the assay (exposure of both populations) resulted in suppression of the response at higher concentrations. A possible mechanism for the enhancement of the MLR by UDMH was suggested by further experiments showing that UDMH inhibited prostaglandin E2 production by adherent splenocytes. PMID- 2972363 TI - Enhancement of mixed lymphocyte culture reactions between donor and recipient in B cell chronic leukemia using antibody-coated beads. AB - Matching of donors and recipients in cases of B cell chronic lymphocytic leukemia (CLL) is complicated by the fact that the cancer clone dominates the blood. A case of CLL was characterized for its antigenic phenotype using monoclonal antibodies. Iron beads were coated with the appropriate antibody and used to remove the cancer clone. A mixed lymphocyte culture (MLC) utilizing the purified cells as well as unpurified cells and sheep cell rosetted purified cells was performed with the donor. Using coated beads, the T cell population rose from 8% to 95% in one treatment versus 8% to 42% with sheep cells. MLC data were statistically better with bead-treated cells, and the results demonstrated the importance of this technique for transplantation matching. PMID- 2972364 TI - Mechanism of immune dysfunction associated with minor antigen graft-vs-host disease in mice. AB - A well-characterized murine model of graft-vs-host disease (GVHD) that develops in response to minor histocompatibility antigens was used to study the mechanism of an immunodeficiency syndrome that is associated with GVHD. Lethally irradiated mice were transplanted with a combination of bone marrow and spleen cells from H 2 compatible donors that differed at multiple minor histocompatibility antigens, or from syngeneic donors. Four to 12 weeks later, the humoral responses of transplanted and control mice to the T dependent antigens bacteriophage phiX174 and TNP-sheep red blood cells (TNP-SRBC), and to the T independent antigen TNP Brucella abortus (TNP-BA) were determined. The results demonstrate that mice with GVHD have relatively intact B Cell function and a profound defect in T helper cell function. The immune response to T dependent antigens normalized with repeated immunization. We conclude that immune dysfunction in mice with GVHD is due to a reversible defect in T helper cell function. PMID- 2972365 TI - Importance of MHC antigen expression on solid tumors in the in vitro interaction with autologous blood lymphocytes. AB - In 54 patients with lung and 14 with ovarian carcinomas the quantitative variations of the major histocompatibility complex (MHC)-determined class I and class II antigens of the tumor cells were related to their in vitro interaction with blood lymphocytes, to the lymphoid infiltration of the tumors, and to the metastatic state of the disease. The tumor cell-lymphocyte interaction was measured by the proliferative response in mixed lymphocyte tumor cell culture and by the cytotoxic activity of the lymphocytes. The results showed that (1) none of the 23 tumors from patients with disseminated disease were lysed; (2) class I negative tumors were not damaged; (3) lymphoid infiltration was present in a higher proportion of class II-positive tumors; and (4) both MHC-positive and negative tumors were found among the disseminated tumors. The requirement of class I expression in the lytic interaction substantiates our earlier conclusion concerning the cytotoxic T lymphocyte nature of lymphocyte-mediated auto-tumor lysis. The lack of auto-tumor lysis in patients with metastases suggests impairment of lymphocyte function in advanced stages of the disease. PMID- 2972367 TI - Coronary angioplasty. PMID- 2972368 TI - Multivessel coronary artery disease. PMID- 2972369 TI - PTCA following coronary bypass surgery. PMID- 2972366 TI - Studies on cytotoxicity generated in human mixed lymphocyte culture. IV. Interleukin 2 alone or from mixed lymphocyte culture yields natural killer-like cytotoxic cells distinct from allospecific cytotoxic T lymphocytes. AB - High levels of cytotoxic activity against the natural killer (NK) cell-sensitive target K562 and the NK-resistant target UCLA-SO-M14 (M14) can be generated in vitro either by mixed lymphocyte culture (MLC) or by culture of lymphocytes in interleukin 2 (IL2) (lymphokine activated killer (LAK) cells). The purpose of this study was to identify similarities and differences between MLC-LAK and IL2 LAK cells and allospecific cytotoxic T cells. Induction of cytotoxicity against K562 and M14 in both culture systems was inhibited by antibodies specific either for IL2 or the Tac IL2 receptor. Like NK effector cells, the precursors for the MLC-LAK cells were low density large lymphocytes. However these precursors differed from the large granular lymphocytes that mediated NK cytolysis in sensitivity to the toxic lysosomotropic agent L-leucine methyl ester (LME). The resistance of the MLC-LAK precursors to LME indicated that the precursors included large agranular lymphocytes. Although interferon-gamma (IFN-gamma) is produced in MLC and in IL2 containing cultures, it is not required for induction of either type of cytotoxic activity. Neutralization of IFN-gamma in MLC- and IL2 containing cultures with specific antibodies had no effect on the induction of cytotoxic activities. Both allospecific cytotoxic T lymphocyte (CTL) and LAK activities were enhanced by IL2 and IFN-gamma at the effector cell stage. However, the mechanism of cytolysis was different in the two systems. NK- and MLC induced LAK activities were independent of CD3-T cell receptor complex while CTL activity was blocked by monoclonal antibodies specific for the CD3 antigen. These results suggest that NK and the in vitro induced LAK cytotoxicities are a family of related functions that differ from CTL. Furthermore, MLC-induced and IL2 induced cytotoxicities against K562 and M14 appear to be identical. PMID- 2972370 TI - Unstable angina and acute myocardial infarction. PMID- 2972371 TI - Vasospastic angina. PMID- 2972372 TI - Total coronary artery occlusion. AB - Although once believed to be unsuitable for coronary angioplasty, patients with subacute and chronic total occlusions have now emerged as important candidates for this procedure. They account for between 10% and 15% of current angioplasties. With careful attention to patient selection factors (occlusion duration, length, morphology, associated thrombus) and technique (initial use of soft guidewires and low-profile dilatation catheters, confirmation of correct wire position, examination for unsuspected distal disease, prolonged heparinization post-procedure), primary success can be achieved in more than 75% of patients. New equipment and techniques--such as thermal or specialized mechanical angioplasty--may facilitate successful crossing of the remaining 25% of total occlusions. The presence of collateral blood flow to the distal vessel provides an important "safety net" and explains the low incidence of emergency surgery following PTCA in this patient group. Patients undergoing successful PTCA of total occlusions should be watched carefully, given the somewhat higher incidence of abrupt reclosure and late restenosis compared with dilatation of conventional stenoses. PMID- 2972373 TI - The high-risk patient. PMID- 2972374 TI - Laser angioplasty: peripheral and coronary applications. PMID- 2972375 TI - Noninvasive assessment of patients following coronary artery angioplasty. AB - PTCA alters the natural history of coronary artery disease and thus presents a challenge for the cardiologist to detect the variety of changes that may occur. Continuous electrocardiographic monitoring with automatic detection of ST segment changes has proved valuable during the immediate post-PTCA procedure. Radionuclide ventriculography and thallium-201 perfusion scans are most important in providing physiologic evidence of improved functional blood flow, particularly in patients with limited revascularization. Finally, the important unsolved problem with PTCA, that of restenosis, is best detected by repeat noninvasive testing in the first months after successful PTCA. PMID- 2972376 TI - Complications of coronary artery angioplasty. PMID- 2972377 TI - Restenosis following coronary artery angioplasty: patterns, recognition, and results of repeat angioplasty. AB - Restenosis is an important problem confronting PTCA. Controversy relative to the long-term outcome of PTCA continues to be fueled by this important problem. At present, certain patterns related to clinical, anatomic, and procedural considerations can be identified as related to restenosis. No interventions based on data from controlled trials offer conclusive treatment to substantially prevent restenosis risk. PMID- 2972378 TI - The long-term followup results of coronary artery angioplasty. AB - The long-term efficacy of PTCA for the relief of angina pectoris and myocardial ischemia has been firmly established. Coronary angiography reveals sustained anatomic improvement in the majority, especially if PTCA is repeated in those patients who developed restenosis. Successful PTCA is associated with a low incidence of cardiac events such as myocardial infarction, elective cardiac bypass surgery, or death during late followup. The majority of patients are completely free of angina and demonstrate objective improvement with exercise testing. Patients with multivessel disease, if properly selected, may derive nearly as much long-term benefit from PTCA as patients with single-vessel disease. PMID- 2972379 TI - Myocardial revascularization: CABG or PTCA? PMID- 2972380 TI - Adjunctive pharmacologic treatment. PMID- 2972381 TI - Protection of the ischemic myocardium during coronary angioplasty. PMID- 2972382 TI - Double loop guiding catheter: a primary catheter for angioplasty of the right coronary artery. AB - Double loop guiding catheters have been used for percutaneous transluminal coronary angioplasty (PTCA) of the right coronary artery (RCA) in 42 consecutive cases. A catheter with a 90-degree primary curve was used when the proximal RCA had horizontal or inferior orientation. When the proximal segment of the RCA was oriented superiorly (shepherd's crook), the catheter with a 75-degree primary curve was used. Catheters were fabricated with short (1.5 cm) or long (2.3 cm) (USCI, C.R. Bard, Inc., Billerica, MA) distal tips. Short-tip catheters were satisfactory in the majority of cases. When the RCA had a complex course and more backup was necessary or when the ascending aorta was wide, long-tip catheters were found to be the best choice. Angioplasty of 49 lesions was attempted in 42 consecutive patients. In 39 patients successful dilatation was achieved (93%). In three patients the procedure was unsuccessful. In one patient, the lesion could not be crossed with the guidewire despite an excellent backup. In another patient, two of three stenoses were dilated successfully; the distal lesion was crossed with a guidewire but could not be crossed with the balloon catheter in spite of a good backup. The lack of a satisfactory engagement and inadequate backup were responsible for the failure in only one patient. There were no complications related to these guiding catheters. We conclude that double loop guiding catheters are safe and can be the primary choice in all right coronary angioplasties. These catheters provide an excellent backup with consequent high success rate. PMID- 2972383 TI - Percutaneous entry of the brachial artery for transluminal coronary angioplasty. AB - Angioplasty is most commonly performed with catheters introduced percutaneously into the femoral artery. A brachial approach has previously been described that employs arteriotomy and cutdown. We describe here the technique and initial results of an alternative method of brachial entry, wherein the catheter is introduced percutaneously. Angioplasty has been performed in 26 patients in this manner, without complications. PMID- 2972384 TI - An unusual case of guide wire fracture during percutaneous transluminal coronary angioplasty. AB - We report a case of guide wire fracture during percutaneous transluminal coronary angioplasty, using one of the newer catheter systems. The broken free end of the guide wire remained within the coronary tree, and surgical removal was necessary. Entrapment and overcoiling of the guide wire can cause fracture; in our case excessive bending in a tortuous coronary tree resulted in this unusual complication. An excessive tensile load to the guide wire may result from anatomical peculiarities and can cause wire fracture. PMID- 2972385 TI - Integration host factor: a protein for all reasons. PMID- 2972386 TI - Two species of human Fc epsilon receptor II (Fc epsilon RII/CD23): tissue specific and IL-4-specific regulation of gene expression. AB - The Fc epsilon receptor II (Fc epsilon RII, CD23) functions in B cell growth and differentiation and in IgE-mediated immunity. The Fc epsilon RII structure expressed on various cell types has been analyzed identifying two species, Fc epsilon RIIa and Fc epsilon RIIb. Sequence analysis of the cloned cDNAs revealed that they differ only at the N-terminal cytoplasmic region, but share the same C terminal extracellular region. These Fc epsilon RII species appear to be generated utilizing different transcriptional initiation sites and alternative RNA splicing. Fc epsilon RIIa is constitutively expressed only in normal B cells and B cell lines, whereas Fc epsilon RIIb expression is detectable in various cell types, such as monocytes and eosinophils. Normally, Fc epsilon RIIb is undetectable in B cells and monocytes, and can be induced by interleukin-4. Moreover, Fc epsilon RIIb is expressed on peripheral blood lymphocytes in atopic individuals. These findings may explain the difference in Fc epsilon RIIa and Fc epsilon RIIb function in B cells and the effector phase of IgE-mediated immunity. PMID- 2972387 TI - Interleukin-1-independent activation of human T lymphocytes stimulated by anti CD3 and a Hodgkin's disease cell line with accessory cell activity. AB - Antibodies directed against the human T cell receptor or the closely associated CD3 molecule stimulate polyclonal T cell proliferation via mechanisms that mimic a primary immune response. We have investigated the requirement for IL-1 production in anti-CD3 (OKT3)-mediated mitogenesis using a Hodgkin's disease cell line (L428) as the accessory cell. L428 cells did not produce detectable IL-1 following stimulation with lipopolysaccharide or phorbol ester (PMA), nor did they transcribe detectable levels of mRNA for IL-1 alpha or beta after such treatment. Despite their inability to produce IL-1, as few as 1 X 10(4) L428 cells reconstituted the proliferative response of accessory cell-depleted T cells to anti-CD3. Although larger numbers of non-rosette-forming (E-) cells were required for maximal responsiveness to anti-CD3, the maximal degree of proliferation was higher with E- cells than with L428 cells. L428-mediated T cell proliferation did not result from residual accessory cells in the responding population or an allogeneic effect since L428 cells were also capable of providing accessory cell activity for the anti-CD3-dependent generation of IL-2 by the Jurkat T cell line. Although the mechanism by which L428 cells provide accessory functions remains incompletely characterized, the ability of anti-HLA DR F(ab')2 fragments to completely abrogate L428 and monocyte-mediated anti-CD3 mitogenesis, despite the addition of exogenous IL-1, provides evidence for the participation HLA-DR molecules in this response. These data indicate that anti CD3-induced proliferation of unprimed human T lymphocytes can occur independently of IL-1 production by accessory cells and may involve the participation of HLA-DR molecules. PMID- 2972388 TI - IL-1 secretion and membrane IL-1 expression by neonatal spleen cells during soluble antigen presentation. AB - Antigen presentation and IL-1 production by neonatal spleen cells were studied in a murine model. The T-helper-cell line (D10-G4.1) (D10), which is specific for soluble antigen presented on syngeneic antigen-presenting cells and dependent on IL-1 for its proliferation, was used as an indicator cell for the ability of syngeneic neonatal or adult spleen cells to present antigen and produce IL-1. The antigen-presenting capacity of neonatal spleen cells is low as attested by D10 proliferation. During antigen presentation there is an augmentation of IL-1 production by the antigen-presenting spleen cell population. However, neonatal spleen cells do not respond to the same levels as do adult spleen cells. These reduced levels of secreted IL-1 cannot be attributed to a low potential for producing IL-1 as attested by the high levels of IL-1 made by these cells after induction by a crude IL-1 inducer factor (IL-1-IF) and by the stimulus of the IL 1-IF produced by D10 cells during antigen presentation by paraformaldehyde-fixed adult cells. The spontaneous expression of membrane IL-1 by neonatal cells is low. Membrane IL-1 levels on neonatal cells can be brought to adult levels by induction with IL-1-IF. Neonatal spleen cells have an impaired capacity to process and/or present soluble antigen. This impairment leads to a decreased stimulus of the T helper cell to produce inducer factors and thus a reduced level of IL-1 production by the neonatal cells during antigen presentation. PMID- 2972389 TI - Suppression of lymphocyte alloreactivity by early gestational human decidua. I. Characterization of suppressor cells and suppressor molecules. AB - We examined the immunosuppressor role of the first trimester human decidua on lymphocyte alloreactivity in vitro in order to identify (1) the major cell classes in the decidua mediating the suppressor effect; (2) the stages in the lymphocyte alloreactive responses susceptible to the suppressor effects of the decidua; and (3) the precise nature of the suppressor molecules. Irradiated (2800 R), Ficoll-Paque-separated nucleated cells of the collagenase-dispersed early gestational (6.5-9.5 weeks menstrual age) decidua containing 70-94% typical decidual cells (identified on the basis of distinctive morphology and numerous cytoplasmic or surface markers) or their plastic-nonadherent fractions further enriched for decidual cells (approximately 96% pure) caused a strong dose dependent suppression of the one way mixed lymphocyte reaction (MLR, i.e., proliferative response measured on Days 3, 4, or 5), when added at the onset of the mixed lymphocyte cultures (MLC). As few as 10(3) decidual cells caused a detectable inhibition of the MLR exhibited by 10(5)-1.5 X 10(5) responder lymphocytes. A smaller degree of suppression was noted with the plastic-adherent fractions of the early decidua (which retained all macrophages and granulocytes, but still included many decidual cells) or unfractionated cells of later gestational (10-13 weeks) decidua containing a higher incidence of leukocytes, granulocytes, and macrophages in particular, or the plastic-adherent fraction thereof, enriched for macrophages. Thus, decidual cells seem to represent an important suppressor cell class in the early gestational human decidua; however, suppression by decidual leukocytes, macrophages in particular, was also evident. The suppressor effect was unrelated to the major histocompatibility phenotype of the responder or the stimulator cells. It was not caused by cell crowding, since an equivalent number of irradiated K562 erythroleukemia cells had little effect on the MLR. The effect was exerted during both the initiation and the progression of the MLR. A delay in the addition of regulator cells progressively minimized the effect on the Day 4 MLR, but did not abolish it completely even when added as late as on Day 3. The major class of mediator molecules was identified as prostaglandins, primarily PGE2, on the basis of the following results: (1) the presence of indomethacin (10(-5) M) or varying dilutions of an anti-PGE2 antibody abrogated this suppression substantially or completely. (2) Addition of pure PGE2 (3 X 10(-7) to 1.1 X 10(-5) M), but not PGF2 alpha, reproduced a dose-dependent suppressor effect.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2972390 TI - A role for L3T4+ T cells and their lymphokines in the generation of suppressor effector (TS3) cells. AB - The cellular requirements for the in vitro induction of antigen-specific suppressor T cells were examined. Previous reports indicated that Ia-bearing macrophages and anti-idiotypic B cells are required as accessory cells to facilitate the generation of suppressor effector (TS3) cells which regulate the response to the 4-hydroxy-3-nitrophenyl acetyl (NP) hapten. The present study describes two distinct T cell populations which interact to generate antigen specific TS3. Fractionation of the T cell populations with monoclonal antibody to the L3T4 determinant led to the identification of an NP-specific L3T4- TS3 progenitor population and an L3T4+ helper/inducer subset. In the presence of NP coupled antigen, the L3T4+ subset could induce progenitor TS3 to differentiate into mature TS3 cells. The activity of the L3T4+ inducer population could be replaced with specifically activated cloned helper cells which were not NP reactive since an I-Ab-restricted, insulin-reactive, L3T4+ clone was capable of supporting the generation of NP-specific TS3. Inducer activity appeared to be confined to the Th1 but not the Th2 subset. In addition, 18-hr supernatants from antigen-activated clones were capable of substituting for L3T4+ cells or T cell clones in TS3 induction cultures. The TS maturation/differentiation factor(s) active in these supernatants does not appear to be IL-1, IL-2, IL-3, or interferon-gamma alone since purified sources of these lymphokines failed to induce TS3 activity. PMID- 2972391 TI - Contributions of host and donor T cells to the inflammatory process in murine lymphocytic choriomeningitis. AB - The severe inflammation characteristic of the infection of adult mice with murine lymphocytic choriomeningitis virus (LCMV) is induced earlier in unsuppressed, virus-infected recipients by the adoptive transfer of class I MHC-compatible, CD4 CD8+ LCMV-immune spleen cell populations. The time to onset of fatal LCM may also be slightly diminished, though not to the extent that would be expected from the enhanced kinetics of the extravasation of cells into cerebrospinal fluid. The development of symptoms is thus not solely related to the magnitude of the inflammatory process. The majority of the T lymphocytes in the inflammatory exudate are of host origin and have the size characteristics of resting cells, while the minority population of donor T cells show more of a lymphoblast morphology. The findings are consistent with the idea that relatively few CD8+ virus-immune effectors trigger an inflammatory process which consists largely of secondarily recruited host T cells and monocyte/macrophages. PMID- 2972392 TI - Regulation of immune responses by T suppressor cells and by serum in chronic paracoccidioidomycosis. AB - Regulation of cellular responses was studied during the course of chronic murine disseminated paracoccidioidomycosis. Regulation of peripheral blood lymphocyte (PBL) proliferative responses to concanavalin A (Con A) was studied in vitro by mixing PBL from infected and noninfected mice. PBL from mice infected for 18 weeks had depressed responses to Con A and they depressed the Con A responses of PBL from noninfected mice by 95% when they were mixed in a 1:1 ratio. After treatment of PBL from infected mice with anti-Lyt-2.2 antibody plus complement, the responses to Con A were increased to normal values. The percentage of T-cell subpopulations in PBL from infected mice did not differ significantly from those of normal mice. Immunoregulation of delayed-type hypersensitivity (DTH) responses to antigen by serum from infected animals was studied in mice 1 week after intranasal (i.n.) infection, a time when DTH responses were maximal. DTH responses to antigen 7 days after i.n. infection (10(7) CFU Paracoccidioides brasiliensis) were significantly reduced when 0.5 ml of immune mouse serum (ELISA antibody titer to P. brasiliensis antigens 1:10,240) was given i.v. 1 day before infection (P less than 0.01) or 1 day before skin testing (P less than 0.001). Normal mouse serum did not have this effect. The results indicate that progression of chronic disseminated paracoccidioidomycosis was associated with the development of T-cell suppressor activity for Con A responses of PBL, and that DTH responses to antigen were depressed by the administration of serum with specific high titer antibodies. PMID- 2972394 TI - IL-4, but not IL-5, can act synergistically with B cell activating factor (BCAF) to induce proliferation of resting B cells. AB - B cell activating factor (BCAF) was initially identified in the supernatant of the murine T helper cell clone 52-3 (52-3 SN) because of its ability to promote activation and proliferation of resting B cells in the absence of any other costimulus. In this paper, we show that 52-3 T helper cells also secrete IL-4 and IL-5 and we have analyzed the influence of these two lymphokines on B cell proliferation induced by BCAF-containing 52-3 SN. Using the neutralizing anti-IL 4 monoclonal antibody 11B11, we observed partial inhibition of B cell proliferation. 52-3 SN free of IL-4 prepared using an immunoabsorbent column was still able to induce significant B cell proliferation. Although recombinant IL-4 alone does not induce B cell proliferation, it increased the proliferation induced by IL-4-free 52-3 SN. Kinetic studies showed that IL-4 is required at the start of B cell cultures in order to exert optimal synergistic effects. In contrast, anti-IL-5 monoclonal antibody NC17 did not affect the B cell proliferative activity of 52-3 SN whether or not IL-4 was present. When 52-3 SN was tested on dextran-sulfate-activated B cells, IL-5 and BCAF activities were detected but only the IL-5 activity was neutralized by monoclonal antibody NC17. These results demonstrate that (i) BCAF-containing SN can induce proliferation of resting B cells independently of IL-4 and IL-5, and (ii) IL-4, but not IL-5, can act synergistically with BCAF to induce B cell proliferation. PMID- 2972393 TI - Synergistic interaction between anti-CD3 and IL-2 demonstrated by proliferative response, interferon production, and non-MHC-restricted killing. AB - Anti-CD3 monoclonal antibody acts on normal peripheral blood mononuclear cells to induce T cell proliferation, interferon-gamma production, and non-MHC-restricted cytotoxicity against both NK (CD16+)-sensitive and -resistant target cells. Moreover, anti-CD3 and interleukin 2 (IL-2) act synergistically to give greater proliferative, interferon-gamma (IFN-gamma), and natural cytotoxicity responses than those expected by the simple addition of the individual responses to each stimulus acting alone. This synergistic response is macrophage independent, greatest at low concentrations of anti-CD3, inhibited by anti-IL2 receptor, and depends upon the induction of IL-2 receptors by CD3 activation which are then available to respond to exogenously added IL-2. Natural cytotoxicity induced by anti-CD3 and IL-2 correlates with IFN-gamma production, is inhibited by anti-IFN gamma, and is still present after depletion of CD16-positive cells by specific monoclonal antibody and complement. The use of anti-CD3 in concert with IL-2 may be worthy of examination in a clinical setting, presumably because CD3/IL-2 generated LAK effector cells could be followed by in vivo administration of potentially lower and less toxic quantities of IL-2 than have been used in the past. PMID- 2972395 TI - Soluble factors in tolerance and contact sensitivity to DNFB in mice. VIII. Regulation of T suppressor cell function by autoreactive T helper cells. AB - When cultured with DNP-labeled I-A+ cells, Lyt 2+ T suppressor cells (Ts) from 2,4,-dinitrobenzene sulfonate (DNBS)-tolerized mice are activated to synthesize and release a suppressor factor (SSF) which suppresses the transfer of contact sensitivity to DNFB. The signals required to activate the DNBS-primed Ts to produce SSF were studied in greater detail. As previously observed with fixed DNP labeled spleen cell stimulators, the supernatants from cultures of DNBS-primed spleen cells and glutaraldehyde-fixed DNP-labeled P388D1 cell monolayers did not contain SSF. When the tolerant cells were harvested from these monolayers and were treated with IL-1, the Ts released the synthesized SSF. Synthesis and release of SSF required Ts recognition of DNP/class I MHC on the hapten presenting cells followed by interaction with the costimulator IL-1. When the tolerant cells were cultured with fixed DNP-labeled I-A+ or I-A- stimulators to induce SSF synthesis, release was induced by adding either unlabeled or TNP labeled unprimed spleen cells to the cultures. The release of SSF was blocked when the second stimulators were pretreated with anti-I-A antibody but not with anti-DNP or anti-class I MHC antibodies. These results indicate that the release of SSF by DNBS-primed Lyt 2+ Ts is regulated by the activity of a self-I-A reactive (i.e., autoreactive) T cell in the tolerant spleen cell population. PMID- 2972396 TI - Neonatal tolerance induction to Mlsa. I. Tolerance to Mlsa is restricted by shared MHC determinants. AB - In this paper we have examined the influence of MHC (major histocompatibility complex) on neonatal tolerance to Mlsa (minor lymphocyte stimulating). By employing a novel approach we have shown that tolerance to Mlsa is restricted by shared MHC determinants. Thus, neonatal Mlsb mice, injected at birth with spleen cells from Mlsa mice, were tested as adults for Mlsa specific responses by interleukin-2 limiting dilution analysis, a technique which allows us to discriminate between responses to MHC + Mlsa and to MHC alone. Tolerance to Mlsa was in the context of any MHC type examined--donor, host, and third-party MHC products. These results show that tolerance to Mlsa is restricted by shared MHC determinants and extend previous studies indicating that activation of Mlsa responses is similarly restricted. PMID- 2972397 TI - Role of the CD45 (T-200) molecule in anti-CD3-triggered T cell-mediated cytotoxicity. AB - NK-depleted human peripheral blood lymphocytes can be modulated with anti-CD3 to kill certain targets during 3-hr cytotoxicity assays. When triggered by anti-CD3 antibody, these effector T cells killed only NK-sensitive targets, such as K562 and HEL 92.1.7, and NK-resistant targets, such as Daudi, whose killing is inhibited by anti-CD45 (T-200) monoclonal antibodies, such as 13.3. NK-sensitive targets, MOLT-4, U266/AF10, Jurkat, and CCFR-CEM, and 10 NK-resistant cell lines, including Raji, IM-9, U698, U937, and GM-1056, whose killing is not inhibited by anti-CD45 monoclonal antibodies, were not killed by alpha-CD3-T effectors, suggesting that the CD45 molecule may be involved in the killing process. Anti CD3-triggered T cell killing of target cells was inhibited greater than 95% by the monoclonal antibody 13.3. This inhibition of cytotoxicity by 13.3 was not due to competition of this IgG1 antibody for Fc receptor binding site on the target cell, since the IgG1 monoclonal antibody anti-beta 2-microglobulin did not block cytotoxicity. Single cell assays and calcium pulse assays showed that CD45 is involved in a postbinding, pre-calcium-dependent stage, similar to that shown for NK cytotoxicity. There was a relative shift of importance of different epitopes of CD45 in anti-CD3-T cytotoxicity compared to NK cytotoxicity. Anti-CD45 antibodies which bind to the C terminus end of the molecule played a more important role in anti-CD3-T cytotoxicity than NK cytotoxicity. Thus, a subset of T cells exists that exhibits anti-CD3-triggered non-MHC-restricted killing of certain NK-sensitive and NK-resistant targets in association with a CD45 molecule which is functionally different from the NK CD45 molecule. PMID- 2972398 TI - Regulation of the distribution of carotenoid droplets in goldfish xanthophores and possible implication to secretory processes. AB - In goldfish xanthophores, the formation of pigment aggregate requires: 1) that a pigment organelle (carotenoid droplet) protein p57 be in the unphosphorylated state; 2) that self-association of pigment organelles occur in a microtubule independent manner; and 3) that pigment organelles via p57 associate with microtubules. In the fully aggregated state, the pigment organelles are completely stationary. Pigment dispersion is initiated by activation of a cAMP dependent protein kinase, which phosphorylates p57 and allows pigment dispersion via an active process dependent on F-actin and a cytosolic factor. This factor is not an ATPase, and its function is unknown. However, its abundance in different tissues parallels secretory activity of the tissues, suggesting a similarity between secretion and pigment dispersion in xanthophores. The identity of the motor for pigment dispersion is unclear. Experimental results show that pigment organelles isolated from cells with dispersed pigment have associated actin and ATPase activity comparable to myosin ATPase. This ATPase is probably an organelle protein of relative molecular mass approximately 72,000, and unlikely to be an ion pump. Isolated pigment organelles without associated actin have 5x lower ATPase activity. Whether this organelle ATPase is the motor for pigment dispersion is under investigation. The process of pigment aggregation is poorly understood, with conflicting results for and against the involvement of intermediate filaments. PMID- 2972399 TI - Dynamics of microtubules visualized by darkfield microscopy: treadmilling and dynamic instability. AB - Individual microtubules undergoing treadmilling in vitro were visualized by darkfield light microscopy, and the relationship between treadmilling and dynamic instability was studied as a function of microtubule-associated proteins (MAPs). In order to demonstrate treadmilling directly by real-time observation, we constructed three-block microtubules, the center-block of which was decorated with Tetrahymena dynein. The decorated block can easily be distinguished from undecorated blocks in the darkfield microscope because the decorated one appears much thicker. At steady-state conditions, the length of an undecorated block at one end increased and that at another end decreased, while the decorated center block did not change in its length. The results from these direct observations show that calf brain 3X-microtubules exhibit a treadmilling flux of 0.9 micron/h. Using a similar microscopy technique, we previously demonstrated that phosphocellulose PC-microtubules existed in either the growing or the shortening phase and alternated quite frequently at steady-state conditions (dynamic instability). How does treadmilling relate to dynamic instability? An image recording of individual 3X-microtubules containing MAPs revealed that the microtubules undergo treadmilling and do not exhibit any dynamic instability. This evidence shows that MAPs suppress the dynamic instability of microtubules. That is, treadmilling can take place in the steady state only after microtubules have been stabilized by MAPs. PMID- 2972400 TI - Dynein as a microtubule translocator in ciliary motility: current studies of arm structure and activity pattern. AB - The dynein arms of ciliary doublet microtubules cause adjacent axonemal doublets to slide apart with fixed polarity. This suggests that there is a unique mechanochemistry to the dynein arm with unidirectional force generation in all active arms and also that not all arms are active at once during a ciliary beat. Negative stain and thin-section images of arms in axonemes treated with beta, gamma methylene adenosine triphosphate (AMP-PCP) show a consistent subunit construction where the globular head of the arm interacts with subfiber B of doublet N+1. This interpretation differs from that provided by freeze etch and STEM interpretations of in situ arm construction and has implications for the mechanochemical cycle of the arm. A computer model of the arms in relation to other axonemal structures has been constructed to test these interpretations. Attachment of the head of the arm subfiber B is directly demonstrable in splayed axonemes in AMP-PCP. About half of the doublets in an axoneme show such attachments, while half do not. This might imply that about half the doublets in an axoneme are active at any given instant and can be identified as such. This information may be useful in probing questions of how active arms differ biochemically from inactive arms and of how microtubule translocators in general become active. PMID- 2972401 TI - [Operative care for abdominal muscle to replace quadriceps femoris musculus without fixation with gypsum]. PMID- 2972402 TI - [Status of estrogen receptors in breast cancer and prediction of chemotherapy sensitivity]. PMID- 2972403 TI - [Photodynamic effect of hematoporphyrin derivative (HpD) plus hyperthermia on human tumor cell lines in culture]. PMID- 2972405 TI - [Electron microscopic observation on human bladder cancer tissue after photodynamic therapy]. PMID- 2972404 TI - [Campylobacter pyloridis and gastroduodenal inflammation]. PMID- 2972406 TI - [Comparative evaluation of pharmacological effects of 9 Scopolia alkaloid quaternaries]. PMID- 2972407 TI - [HbD Ouled Rabah [beta 19 (B1) Asn----Lys] as a rare beta-chain detected by reverse phase high performance liquid chromatography]. PMID- 2972408 TI - [Observation on tumor invasion in vivo after subcapsular transplantation of a mouse forestomach carcinoma into a syngenic mouse kidney]. PMID- 2972409 TI - [Memory enhancement of [Arg8] vasopressin (AVP): effect and mechanism. I. Propranolol blocking effect on memory retention of AVP in mice]. PMID- 2972410 TI - [Expression of the IL-2 gene and its response to heat shock]. PMID- 2972411 TI - [Observation on blood biochemical indices after gossypol treatment in gynecological cases]. PMID- 2972412 TI - [Studies of distribution and myocardial imaging on 99mTc-CPI in animals]. PMID- 2972413 TI - [Nutritional status of vitamin A and carotenes in miners and lung cancer patients]. PMID- 2972414 TI - [Simultaneous determination of lidocaine and quinidine in serum by a simple HPLC method]. PMID- 2972415 TI - [Rapid detection of protein and nucleic acid homology with a microcomputer]. PMID- 2972416 TI - Modulation of natural killer activity by 12-O-tetradecanoylphorbol-13-acetate and benzoyl peroxide in phorbol ester-sensitive (SENCAR) and resistant (B6C3F1) mice. AB - Following two weeks of topical application of 12-O-tetradecanoylphorbol-13 acetate (TPA) at 2, 4 and 8 micrograms/mouse on alternate days (7X total) or benzoyl peroxide (BZP) at 10, 20 and 40 mg/mouse, natural killer (NK) activity was determined in local (lymph nodes draining the lower dorsal region) and systemic (spleen) lymphoid tissue in phorbol ester-sensitive (SENCAR) and resistant (B6C3F1) mice. SENCAR mice, sensitive to tumor induction by TPA in two stage chemical-induced carcinogenesis protocols, demonstrated suppression of NK activity in the spleen (no significant change in lymph nodes) and substantial dose-dependent increases in cell numbers in these organs after topical exposure to TPA. B6C3F1 (C57BL/6 X C3H F1) mice, reported to be resistant to TPA-induced promotion, demonstrated significant increases in NK activity in lymph nodes/spleen with an increase in cell numbers in the draining nodes only. Unlike the C57BL/6 parental strain, B6C3F1 mice are also reported to be resistant to promotion with BZP. Significantly, studies in this laboratory indicated that B6C3F1 mice dosed with BZP demonstrated increased NK activity in the spleen as was observed after dosing with TPA. These data suggest that alterations in NK activity as a result of exposure to tumor promoters may, in part, account for the resistance of particular strains of mice to tumor development. In both SENCAR and B6C3F1 mice, the blastogenic response of spleen cell suspensions isolated from TPA-dosed animals to phytohemagglutinin (PHA), a T cell lectin, was suppressed in a dose-dependent manner; BZP had no effect on spleen cell responses in either strain. Blastogenic responses of lymph node cells to PHA were enhanced in both strains of mice after topical application of TPA and BZP. Therefore, alterations in lymphoid cell responsiveness to PHA appeared unrelated to the reported sensitivities of SENCAR and B6C3F1 mice to tumor promotion. PMID- 2972417 TI - Long-term serial changes in left ventricular function and reversal of ventricular dilatation after valve replacement for chronic aortic regurgitation. AB - In most patients with aortic regurgitation, valve replacement results in reduction in left ventricular dilatation and an increase in ejection fraction. To determine the relation between serial changes in ventricular dilatation and changes in ejection fraction, we studied 61 patients with chronic severe aortic regurgitation by echocardiography and radionuclide angiography before, 6-8 months after, and 3-7 years after aortic valve replacement. Between preoperative and early postoperative studies, left ventricular end-diastolic dimension decreased (from 75 +/- 6 to 56 +/- 9 mm, p less than 0.001), peak systolic wall stress decreased (from 247 +/- 50 to 163 +/- 42 dynes x 10(3)/cm2), and ejection fraction increased (from 43 +/- 9% to 51 +/- 16%, p less than 0.001). Between early and late postoperative studies, diastolic dimension and peak systolic wall stress did not change, but ejection fraction increased further (to 56 +/- 19%, p less than 0.001). The increase in ejection fraction correlated with magnitude of reduction in diastolic dimension between preoperative and early postoperative studies (r = 0.63), between early and late postoperative studies (r = 0.54), and between preoperative and late postoperative studies (r = 0.69). Late increases in ejection fraction usually represented the continuation of an initial increase occurring early after operation. Thus, short-term and long-term improvement in left ventricular systolic function after operation is related significantly to the early reduction in left ventricular dilatation arising from correction of left ventricular volume overload. Moreover, late improvement in ejection fraction occurs commonly in patients with an early increase in ejection fraction after valve replacement but is unlikely to occur in patients with no change in ejection fraction during the first 6 months after operation. PMID- 2972418 TI - Myocardial protection during coronary angioplasty with an autoperfusion balloon catheter in humans. AB - An autoperfusion balloon catheter was developed to allow passive myocardial perfusion during inflation through a central lumen and multiple side holes in the shaft proximal and distal to the balloon. We report its safety and efficacy in 11 patients undergoing elective angioplasty to a single coronary lesion. Each lesion was dilated three times with the autoperfusion inflation bracketed between two inflations by standard angioplasty catheters. Chest pain score, 12-lead electrocardiogram, heart rate, and mean aortic pressure were recorded before each inflation and at 1-minute intervals after inflation. Inflation duration during autoperfusion angioplasty (513 +/- 303 seconds) was longer than for the pre- (107 +/- 55 seconds, p = 0.0004) and post- (139 +/- 71 seconds, p = 0.0006) standard dilatations. The maximum ST-segment elevation and depression in any lead during autoperfusion angioplasty (0.3 +/- 0.5 and 0.6 +/- 0.8 mm) was significantly less than for the pre- (2.4 +/- 1.7 mm, p = 0.002 and 2.2 +/- 1.3 mm, p = 0.0004) or post- (1.9 +/- 1.3 mm, p = 0.002 and 1.6 +/- 1.3 mm, p = 0.018) standard dilatations at the same point in time. Maximal chest pain score during autoperfusion (3.2 +/- 3.5) was lower than for the pre- (6.1 +/- 2.1, p = 0.003) but not the post- (5.2 +/- 3.1, p = 0.07) standard angioplasty. All 11 patients underwent successful, uncomplicated procedures. We conclude that this autoperfusion catheter significantly reduces ischemic symptoms and signs during coronary angioplasty, allowing prolonged periods of balloon inflation. PMID- 2972419 TI - Coronary vasodilator reserve after human orthotopic cardiac transplantation. AB - Cardiac transplantation is frequently associated with accelerated coronary atherosclerosis and immune-mediated microvascular injury. To determine if orthotopic cardiac transplantation impairs the capacity of the coronary vasculature to vasodilate and conduct hyperemic blood flow, maximal coronary vasodilator reserve was measured in 25 cardiac allograft recipients with no evidence of rejection 6-57 months after transplantation and in 20 normal subjects. Left ventricular wall thickness was assessed echocardiographically, and epicardial coronary anatomy was evaluated by quantitative coronary angiography. Coronary vasodilator reserve (CVDR) was measured in all patients with a coronary Doppler catheter and a maximally vasodilating dose of intracoronary papaverine. CVDR measured in the transplant recipients with normal coronary arteries, left ventricular function, and wall thickness (5.0 +/- 0.3 [mean +/- SEM] peak/resting velocity; range, 3.8-7.3; n = 16) was not different from that of normal subjects (4.8 +/- 0.2; range, 3.7-8.3). CVDR in the five cardiac allograft recipients with diffuse coronary atherosclerosis producing 30 +/- 5% narrowing (range, 25-38%) of epicardial vessel diameter also was normal (5.1 +/- 0.3; range, 4.3-6.2; n = 5). The CVDR was reduced, however, in two of the four cardiac allograft recipients with left ventricular hypertrophy. In the only transplant recipient in whom a regional wall motion abnormality was present, CVDR was abnormal in the vascular distribution of the hypokinetic wall segment (1.8) but was normal in the artery that supplied normally functioning myocardium (4.0). These findings demonstrate that in the absence of allograft rejection, acquired left ventricular hypertrophy, and regional wall motion abnormalities, coronary vasodilator reserve is normal after orthotopic human cardiac transplantation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972420 TI - Applications of percutaneous transluminal coronary angioplasty in cardiac transplantation. Preliminary results in five patients. AB - Atherosclerotic coronary artery disease is the major cause of late cardiac transplant failure secondary to silent ischemia and infarction. To increase the longevity of cardiac homografts, we performed percutaneous transluminal coronary angioplasty (PTCA) in five male patients (aged 45 +/- 7 years, mean +/- SEM); 17 lesions were dilated during eight procedures 83 +/- 11 months after cardiac transplant. PTCA was successful (greater than or equal to 20% change in vessel diameter) in 13 of 17 (76%) lesions (the degree of prePTCA stenosis was 84% +/- 3% vs. 40% +/- 4% postPTCA; p less than or equal to 0.01). Multiple PTCA procedures were performed for progressive coronary artery disease in two patients; in one patient, two procedures were 13 months apart, and, in the second patient, another three procedures were 2 and 6 months apart. Indications for PTCA included reversible thallium perfusion defects, segmental left ventricular wall motion abnormalities, or both in the distribution of proximal coronary artery stenoses. No deterioration occurred in the four unsuccessful PTCA attempts (two patients with initial total occlusion, and two patients in whom the lesion could not be crossed with a guidewire). Noninvasive evidence of ischemia was improved immediately after PTCA in all cases. Three patients remain alive 5, 7, and 11 months, respectively, after PTCA without evidence of new ischemia. One patient died 39 months after his first PTCA, while another patient was retransplanted 8 months after the first PTCA. Thus, PTCA can be performed in cardiac transplant patients with proximal major vessel coronary artery disease and may prolong cardiac homograft function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972421 TI - Enzyme-linked immunosorbent assay of lipoprotein(a) in serum and cord blood. AB - We have developed a new sensitive method for quantifying lipoprotein(a) (Lp(a] in human serum, using a 'sandwich' type noncompetitive enzyme-linked immunosorbent assay (ELISA). The solid-phase used was a polystyrene plate. The anti-Lp(a) antibody-enzyme conjugate was labelled by linking Fab' fragments to peroxidase (EC 1.11.1.7) by the maleimide method. The minimum detectable concentration was 0.5 ng/well. Routinely, the assay was carried out with 1,000-fold diluted serum, and Lp(a) was quantified between 4.0 and 500 mg/l. Within-run coefficients of variation (CVs) ranged from 3.5% to 10.4% and between-run CVs from 5.0% to 11.1%. Results by the ELISA were in good agreement with those by radial immunodiffusion (r = 0.955). The distribution of Lp(a) in serum from 820 healthy donors was highly skewed: mean 141.1 mg/l, medium 97.9 mg/l. In cord blood, the mean and median were 15.6 and 9.8 mg/l, respectively. This ELISA for Lp(a) has the advantages of being highly sensitive and specific, simple to perform, and does not use radioisotopes. PMID- 2972422 TI - Pathophysiology of increased urinary N-acetyl-beta-D-glucosaminidase activity in human hypertension: effect of cilazapril therapy. AB - To investigate the pathophysiology of elevated urinary N-acetyl-beta-D glucosaminidase (NAG) activity in human hypertension, total and fractional urinary NAG activities were correlated with Li clearance, a measure of proximal renal tubular Na reabsorption, in 15 white hypertensive subjects, aged 56 years, and in 13 normotensive control subjects. In 11 hypertensive subjects, the measurements were repeated after 2 months of antihypertensive therapy with the converting enzyme inhibitor cilazapril (2.5 or 5.0 mg daily). Urinary NAG activity was increased in hypertensives and correlated directly with Li clearance in both hypertensive and normotensive subjects. Li clearance and urinary NAG activity were reduced by antihypertensive therapy. The findings indicate an inverse correlation between urinary NAG activity and proximal renal tubular Na reabsorption, suggesting that NAG is reabsorbed in the proximal renal tubules with Na and other proteins. High Li clearance and urinary NAG activity in hypertension are in part functional and, therefore, reversible disturbances of renal function. PMID- 2972423 TI - Effect of long-term treatment with diltiazem on atrial natriuretic peptides in spontaneously hypertensive rats. AB - The present study was designed to examine the possible effect of long-term treatment with diltiazem on plasma and atrial concentrations of atrial natriuretic peptides (ANP) in spontaneously hypertensive rats (SHR). Diltiazem treatment reduced blood pressure and ventricular weight in SHR. Plasma ANP concentration in untreated SHR was higher than Wistar-Kyoto rats (WKY). Diltiazem treatment decreased plasma ANP concentration in SHR near to the level of WKY; moreover, plasma ANP concentration was correlated with blood pressure and ventricular weight in treated and untreated SHR. Left atrial ANP concentration in untreated SHR was lower than WKY. Diltiazem treatment increased left atrial ANP concentration in SHR, but this effect was not noted in WKY. These results suggest that the ANP release from the left atrium is chronically stimulated in adult SHR, and that the prevention of an increase in plasma ANP by diltiazem treatment may be, in part, attributed to the improvement of cardiac overload induced by reductions in blood pressure and cardiac hypertrophy. PMID- 2972424 TI - Huntington disease carrier status and the problems involved for those affected. A psychotherapeutic experience. AB - The paper presents a case report of a clinically healthy woman with a family history of Huntington disease, whose child developed symptoms of the disease at the age of 8 years. Psychotherapeutic treatment for more than 18 months has provided insight into the problems associated with being an unsymptomatic carrier of the gene and helps to elucidate some aspects of predictive testing. PMID- 2972425 TI - Characterization of lymphocytes that suppress IL-2 production in systemic lupus erythematosus. AB - IL-2 production by peripheral blood mononuclear cells (PBM) is decreased in patients with systemic lupus erythematosus (SLE). This defect can be reversed by the removal of CD8+ lymphocytes. The purpose of these studies was to determine whether the CD8+ IL-2 suppressor cells comprise a specific subset or whether all CD8+ cells have this activity. Lymphocyte subsets were identified and separated by two-colour flow cytometry prior to a 48 h mitogen stimulation. The CD8+ cells that suppressed IL-2 production co-expressed HLA DR and were radiosensitive. Other markers co-expressed by CD8+ cells which are found on suppressor cells such as Leu 15 (CD11), Leu 11 (CD16), and Leu 7 were also found on the CD8+ IL-2 suppressor cell population in SLE. In healthy subjects, removal of CD16+, but not of CD8+ cells markedly elevated the production of IL-2. The CD8- CD16+ non-T cell subset suppressed IL-2 production by normal and SLE PBM in autologous and allogeneic combinations. This subset may be a human equivalent of the murine natural suppressor cells. These results demonstrate that the cells that suppress IL-2 production in SLE are heterogeneous, and suggest that they belong to more than one lineage. PMID- 2972426 TI - Deficient interleukin 2 production in rheumatoid arthritis: association with active disease and systemic complications. AB - Interleukin 2 (IL-2) production and proliferative responses of peripheral blood mononuclear cells (PBMC) stimulated with three concentrations of PHA were measured in 75 patients with rheumatoid arthritis (RA) and 25 normal controls. All patients were on a standard therapeutic regime, and were assessed for disease activity by clinical and laboratory criteria. Rheumatoid cells showed significantly lower IL-2 production and proliferation than normal PBMC at all PHA doses. These differences were not attributable to different kinetics. Within the rheumatoid population, both IL-2 levels and proliferation were lower in patients with active disease than those with inactive RA. Patients with extra-articular disease showed the most pronounced defects. Proliferative responses showed an inverse correlation with clinical indices of disease activity but not with measures of the acute phase response. Rheumatoid patients had higher proportions of CD4+, TFR+ and Tac+ lymphocytes than controls. Both proliferative responses and IL2- levels showed a positive relationship with the proportion of CD4+ cells, and an inverse relationship with Tac+ lymphocytes. Monocyte depletion and partial reconstitution resulted in an increase of both proliferation and IL-2 production, which was more marked in RA patients, suggesting that depressed IL-2 production may relate in part to monocyte effects. However, this cannot completely explain the magnitude of the defects observed, because normal monocytes did not increase the responses of rheumatoid lymphocytes, neither did rheumatoid monocytes suppress the responses of normal lymphocytes. PMID- 2972427 TI - The majority of the activated T cells in the blood of insulin-dependent diabetes mellitus (IDDM) patients are CD4+. AB - Twenty-five recently diagnosed insulin-dependent diabetic patients were screened for the presence of activated T lymphocytes using the anti-IL-2 receptor monoclonal antibody; thirteen had elevated levels of activated T lymphocytes. The activated cells were mostly confined to the CD4 subset, with the CD4/CD8 ratio in IL-2 receptor expressing cells averaging 5 +/- 1 (s.d.) compared with 1.3 +/- 0.3 for all T cells. In recent onset insulin-dependent diabetes blood there was no lack of CD4 CD45R+ (suppressor/inducer) T cells. The activated IL-2 receptor, expressing cells belonged to both CD4 subsets, 'helper inducer' (CD44B4) and 'suppressor inducer', (CD42H4). PMID- 2972428 TI - Distinct phenotypic composition of diffuse interstitial and perivascular focal infiltrates in renal allografts: a morphometric analysis of cellular infiltration under conventional immunosuppressive therapy and under cyclosporine A. AB - Phenotypic analysis of interstitial mononuclear cell infiltrates was undertaken in 40 transplant renal specimens obtained from 38 patients in order to assess the influence of immunosuppressive therapy. Thirteen patients were given conventional immunosuppressive treatment (azathioprine and prednisone) and the other 25 received cyclosporine. The immunostaining was performed using seven antileucocyte antibodies by alkaline phosphatase-anti-alkaline phosphatase method. Interstitial infiltrates were distributed in two patterns: diffuse infiltrates and periglomerular/perivascular aggregates. The phenotypic composition was distinct in these two patterns: in diffuse infiltrates, monocytes/macrophages (EBM 11) represented the predominant inflammatory cell and were associated with a minor component of T cells (T 11). In contrast, aggregates had a major T lymphocyte phenotype in addition with few foci of B cells. T4 subset of T lymphocytes always predominated over T8 subset. The repartition and the proportion of each cell type were not significantly different in rejecting and not rejecting grafts and were not affected by the immunosuppressive regimen. PMID- 2972429 TI - Involvement of atrial natriuretic peptide in long-term regulation of sodium homeostasis in man. PMID- 2972430 TI - Synovitis-acne-pustulosis hyperostosis-osteomyelitis syndrome (SAPHO). A new syndrome among the spondyloarthropathies? PMID- 2972431 TI - Endocrine effects of combined treatment with an LHRH agonist in association with flutamide in metastatic prostatic carcinoma. AB - The plasma levels of pituitary hormones (LH, FSH and prolactin) as well as testosterone were determined in 62 patients treated with combined therapy using the LHRH agonist [D-Trp6, des-Gly-NH2(10)]LHRH ethylamide and the antiandrogen Flutamide. Plasma radioimmunoassayable LH and FSH levels increased to 534% (p less than 0.01) and 150% (p less than 0.01) of control, respectively, during the first 5 days of treatment, while, afterwards, a marked inhibition was observed which remained constant at approximately 30-50% of control values during the whole period of treatment. All patients showed a decrease of plasma testosterone concentration to approximately 10% of control levels. Detailed determinations of plasma testicular and adrenal steroid levels were then performed in 15 patients. Our data indicate that, except for the blockade of testicular 17 hydroxyprogesterone secretion, the combined therapy has no effect on plasma C-21 steroid levels. However, adrenal C-19 steroids, namely dehydroepiandrosterone and its sulfate, androst-5-ene-3 beta, 17 beta-diol and androstenedione were decreased to approximately 50% of control values (p less than or equal to 0.01). The main testicular steroids, testosterone and dihydrotestosterone, which were increased during the first 10 days of combined administration, rapidly decreased and reached approximately 10% of control values at later time intervals. The present study extends our previous observations indicating that the combined antihormonal treatment affects both testicular and adrenal steroidogenesis. Moreover, we have demonstrated that, up to at least 2 years, this treatment, in addition to decreasing the serum levels of testicular androgens, causes an inhibition of the plasma levels of C-19 steroids from adrenal origin. PMID- 2972432 TI - Evaluation of Dacron-covered and plain bovine xenografts as replacements for the anterior cruciate ligament. AB - Surgical repair of the anterior cruciate ligament often involves the use of a suitable autograft. As alternatives to sacrificing these normal structures, various allografts, xenografts, and synthetic materials have been investigated as ligament replacement materials. This study investigates Dacron fabric-covered and plain bovine xenograft tendon as such materials in the canine knee. The implants were tested to failure in an MTS machine following 13 weeks of implantation in a canine knee. Dacron woven fabric-covered implants became more firmly attached than those covered by Dacron mesh fabric or plain xenografts. The implants were also analyzed according to their method of attachment (fixation staples or sutures). Overall, the sutured implants failed at slightly higher forces than did the stapled ones. Histologically, limited vascular invasion of the xenograft was observed. No host fibrous or osseous tissue could be identified within the graft. Fibrous tissues did form between the bone and xenograft. The implants exhibited extreme intraarticular wear, which suggests a low potential for intraarticular ligament replacement. PMID- 2972433 TI - Iliopsoas abscess in an immunocompromised patient. PMID- 2972435 TI - Finding and dilating renal artery stenoses for hypertension. PMID- 2972434 TI - Altered metabolism and decreased efficacy of prednisolone and prednisone in patients with hyperthyroidism. AB - To evaluate the effect of hyperthyroidism on the protein binding and metabolism of prednisolone, eight subjects with hyperthyroidism were investigated before and after thyroid status returned to normal. Hyperthyroidism was associated with a reduced volume of distribution of prednisolone, a decreased systemic availability of prednisolone after oral prednisone, a displacement of the prednisolone in equilibrium with prednisone equilibrium toward prednisone, and an increased nonrenal clearance of unbound prednisolone in the presence of an impaired 6 beta hydroxyprednisolone formation. After oral prednisone or intravenous prednisolone, patients with hyperthyroidism had lower albumin-bound, transcortin-bound, and unbound concentrations of prednisolone but normal affinities of albumin and transcortin for prednisolone binding. These differences in prednisolone plasma concentrations were biologically relevant, because the capacity of these plasma samples to inhibit allogeneically stimulated lymphocytes was lower by 70% in the hyperthyroid than in the euthyroid state. Thus hyperthyroidism reduces the biologic effect of prednisolone and exhibits a differential effect on various enzymes involved in the catabolism of prednisolone. PMID- 2972436 TI - Transluminal balloon dilatation. PMID- 2972437 TI - Grading for angioplasty. AB - An increasing proportion of patients with lower limb arterial disease are being treated by transluminal angioplasty. There have been various estimates of the numbers of potentially treatable patients, but none have been accompanied by detailed selection criteria. Based on patients' arteriograms, we present a clear grading system of suitability for angioplasty aimed at allowing ready interpretation and comparison of data. We have used these grades to examine prospectively the suitability for angioplasty of 118 consecutive patients undergoing arteriography for the first time. Thirty-five per cent of patients with claudication, and 30% of those with rest pain or trophic lesions appeared suitable for angioplasty. Forty-five per cent of claudicants and 40% patients with rest pain or trophic lesions could have their symptoms relieved by appropriate angioplasty of aortoiliac disease alone in the presence of distal disease, or of isolated femoropopliteal lesions. These figures have important implications for the provision of interventional radiological services throughout the United Kingdom. PMID- 2972438 TI - [Long-term therapy of retinal vasculopathies with oral administration of high doses of O-(beta-hydroxyethyl)-rutoside]. PMID- 2972439 TI - [A double-blind controlled clinical study on the use of mucopolysaccharides in the treatment of patients with cerebral vasculopathy. Comparison with a placebo]. PMID- 2972440 TI - [Traumatic contusions of athletes]. PMID- 2972441 TI - [Double-blind parallel comparison of midazolam and triazolam in sleep disorders in the aged]. PMID- 2972442 TI - [Long-term treatment with nifedipine does not alter carbohydrate tolerance in normal subjects and does not worsen metabolic control in diabetics]. PMID- 2972443 TI - [Clinical study of intestinal infections treated with norfloxacin]. PMID- 2972444 TI - [Recent developments in contrast media]. PMID- 2972445 TI - [Drug-induced lupus]. PMID- 2972446 TI - [Hyperglycemic hyperosmolar coma without ketoacidosis during treatment with chlorthalidone]. PMID- 2972447 TI - [How to recognize and treat iodine deficiency which manifests as goiter?]. PMID- 2972448 TI - [Therapeutic advances in bone marrow transplantation]. PMID- 2972449 TI - [Research on the therapeutic activity and tolerance of flunoxaprofen in arthrosis]. PMID- 2972450 TI - [Efficacy of regulators of the intestinal bacterial flora in the therapy of acne vulgaris]. PMID- 2972452 TI - [Incidence of cesarean section in elderly pregnant women]. PMID- 2972451 TI - [Further observations on the effects of the treatment with amitriptyline and perphenazine in essential headache]. PMID- 2972453 TI - [Dynamics of the lower urinary tract after a rapid bolus intake of Fabia water]. PMID- 2972454 TI - [Aromatic amines: pathology and prevention]. PMID- 2972455 TI - [A controlled double-blind clinical study of 15 CH barium carbonate vs placebo in a group of hypertensive inmates of 2 homes for the aged]. PMID- 2972457 TI - [Pregnancy and labor in patients under 17 years of age. Clinico-statistical considerations]. PMID- 2972456 TI - [Use of a new delayed-release anhydrous theophylline by single daily administration in the treatment of reversible bronchial obstruction. Comparison with a delayed-release preparation administered twice daily]. PMID- 2972458 TI - [Free oxygen radicals. Their role in the physiopathology of rheumatic diseases and therapeutic implications]. PMID- 2972460 TI - [Nosography and risk factors in obliterating arteriopathies of the lower limbs]. PMID- 2972459 TI - [Effects of crenotherapy with a salso-bromo-iodine mineral water in hyperuricemic patients]. PMID- 2972461 TI - [Recommendations for surveillance and participation in school of children with HIV seropositivity]. PMID- 2972462 TI - [Effects of coenzyme Q10 on physical exercise tolerance and cardiac performance in normal untrained subjects]. PMID- 2972463 TI - [Short-term sucralfate treatment of duodenal ulcer]. PMID- 2972464 TI - [A new therapeutic approach to iron deficiency]. PMID- 2972466 TI - [Calcium antagonists and cerebrovascular circulation]. PMID- 2972465 TI - [Advances in neurosurgery]. PMID- 2972467 TI - [When and why to start drug therapy in essential hypertension]. PMID- 2972468 TI - [Updating deep venous thrombosis. I. Physiopathology and clinical aspects]. PMID- 2972469 TI - [Clinical picture of HIV]. PMID- 2972470 TI - [Telethermographic evaluation of pirprofen in rheumatic pathology]. PMID- 2972472 TI - [Mainly neutropenic pancytopenia caused by acute bone marrow depression during ticlopidine treatment]. PMID- 2972471 TI - [Treatment of erythrasma with 1% bifonazole cream]. PMID- 2972473 TI - [Current therapeutic aspects of Trichomonas vaginalis infections in gynecology]. PMID- 2972475 TI - [News about AIDS]. PMID- 2972474 TI - [Can thymostimulin influence the course of malignant subacute hepatitis?]. PMID- 2972476 TI - [Therapy of Paget's disease]. PMID- 2972478 TI - [Criteria for the evaluation of the therapeutic use of calcium antagonist drugs]. PMID- 2972477 TI - [Action of calcitonin in peri- and post-menopausal osteoporosis and in osteoporosis induced by long-term treatment with cortisone]. PMID- 2972479 TI - [Efficacy and tolerance of a preparation containing triamcinolone acetonide, nystatin and neomycin]. PMID- 2972480 TI - [A semirigid corset in the treatment of lumbar disc arthrosis. Physiopathology, indications, limitations and counterindications. A clinical contribution]. PMID- 2972481 TI - [Psychiatric disorders and vitamin B12 deficiency. Considerations apropos of a case]. PMID- 2972482 TI - [Amiodarone and thyroid function]. PMID- 2972483 TI - [Diagnostic imaging in the evaluation of jaundice]. PMID- 2972485 TI - [Hyposensitization]. PMID- 2972484 TI - [Diagnostic imaging in the evaluation of hepatic metastasis]. PMID- 2972487 TI - [Use of flunarizine in the therapy of hemicrania. Clinical evaluation in 25 patients]. PMID- 2972486 TI - [Prostaglandins and gastrointestinal cytoprotection]. PMID- 2972488 TI - [Topical treatment of acne with spironolactone]. PMID- 2972489 TI - [Role of lymphapheresis in treatment protocols of rheumatoid arthritis]. PMID- 2972490 TI - [Calcium antagonists and blood filterability]. PMID- 2972491 TI - [2 cases of lidocaine-resistant ventricular tachycardia treated with i.v. mexiletine during complicated acute myocardial infarct]. PMID- 2972493 TI - [Psychological problems in the treatment of obese patients]. PMID- 2972492 TI - [Therapy of erythema nodosum]. PMID- 2972494 TI - [Hypertensive emergencies treated with nifedipine drops]. PMID- 2972495 TI - [Activity and tolerance of a dermatologic preparation with a triamcinolone acetonide and nystatin base in skin diseases caused by Candida albicans]. PMID- 2972497 TI - [Sclerosing therapy of varices. Indications and technical notes]. PMID- 2972496 TI - [Polyunsaturated phosphatidylcholine + vitamin B complex in the therapy of alcoholic hepatopathies. A controlled study versus standard therapy]. PMID- 2972498 TI - [Acute abdominal pain in emergency medicine]. PMID- 2972499 TI - [Clinical features of a case of Dandy-Walker malformation]. PMID- 2972500 TI - [Humeral epicondylitis: notes on therapy]. PMID- 2972501 TI - [Therapeutic possibilities in Rendu-Osler-Weber disease]. PMID- 2972502 TI - [Technic and application of light reflection rheography]. PMID- 2972503 TI - [The grade of dyspnea in patients treated with a theophylline derivative (bamifylline) vs controlled-absorption theophylline]. PMID- 2972504 TI - [Inclusion of flunarizine in the antiepileptic drug protocol. Clinical and laboratory observations]. PMID- 2972505 TI - [Therapeutic use of metadoxine in alcohol-related mental and behavioral disorders]. PMID- 2972506 TI - [Drug therapy of arthrosis]. PMID- 2972507 TI - Papulosquamous eruptions: manifestation and management. PMID- 2972508 TI - Increased mortality in mice infected with rabies virus and subsequently vaccinated against rabies. AB - When mice infected 1 or 2 days before by an IM inoculation after high passage of the virus in the species ("challenge virus standard" strain) received an injection of live (Flury) or inactivated virus, their mortality was increased in comparison with unvaccinated controls. In the case of the inactivated virus vaccine, mortality was proportional to the dose of vaccine received. Conversely, when vaccination was carried out in mice recently infected with the same doses of a heterologous strain adapted to foxes, this phenomenon could not be demonstrated. The consequences of these observations on failures of treatment in animals infected with a homologous strain, cases of rabies occurring after vaccination or quality control of vaccines are discussed. PMID- 2972509 TI - An investigation of the irritant and allergenic properties of daffodils (Narcissus pseudonarcissus L., Amaryllidaceae). A review of daffodil dermatitis. AB - Irritancy of daffodil flowers and bulbs was assessed using various fresh plant preparations, solvent extracts and some of the known Amaryllidaceae alkaloids on guinea pigs. Sensitization was also carried out on guinea pigs using these plant preparations, solvent extracts and 7 fractions obtained after preparative chromatography of the bulb ether extract. Only 1 fraction, containing 2 alkaloids, was capable of inducing delayed hypersensitivity in the animals; the sensitivity achieved, however, was weak. The substances were identified as masonin and homolycorin, which acted as elicitors, but masonin may also be a sensitizer. While homolycorin is a known daffodil constituent, masonin has not been found previously in Narcissus pseudonarcissus. 3 other alkaloids as well as chelidonic acid and isorhamnetin were non-elicitors in the sensitized guinea pigs. PMID- 2972510 TI - Generalized eruption with severe liver dysfunction associated with occupational exposure to trichloroethylene. AB - Generalized dermatitis due to trichloroethylene is rare. A 21-year-old printer developed exfoliative dermatitis with mucous membrane involvement, fever and liver dysfunction after a 2-week occupational exposure to trichloroethylene. Positive patch-test reactions to trichloroethylene and to one of its metabolites, trichloroethanol, were observed. This dermatitis is considered to be mediated by a delayed-type hypersensitivity mechanism. PMID- 2972512 TI - Contact urticaria and anaphylactoid reaction from cornstarch surgical glove powder. PMID- 2972511 TI - Contact dermatitis from nickel: an investigation of its sources. AB - Patch tests with the GEIDC standard series of allergens, and with 8 washers made of copper, nickel, nickel-palladium, palladium, brass, bronze, gold and iron, were carried out in 964 consecutive patients who complained of intolerance to metals and in 200 controls who did not. All subjects were also questioned as to personal and family history of atopy, occupational contact and intolerance to gold. The results provide support for the substitution of nickel in imitation jewelry with metals such as palladium or bronze. PMID- 2972513 TI - Occupational irritant contact dermatitis from fibreboard containing urea formaldehyde resin. PMID- 2972514 TI - Occupational allergic contact dermatitis from Kathon WT. PMID- 2972515 TI - Contact dermatitis from acrylamide. PMID- 2972516 TI - Allergic contact dermatitis from triglycidyl isocyanurate in polyester paint pigments. PMID- 2972518 TI - Contact dermatitis due to glyceryl monothioglycolate. PMID- 2972517 TI - Occupational allergic contact dermatitis from amethocaine. PMID- 2972519 TI - Allergic contact dermatitis from selenite. PMID- 2972520 TI - Patch testing with a mixture of 2 phenol-formaldehyde resins. AB - 1310 patients were routinely patch tested with a paratertiary-butylphenol formaldehyde resin (PTBP-F-R), a resol resin based on phenol and formaldehyde (P F-R-2), and a mixture of these 2 resins. Approximately 2.5 times more patients with contact allergy to phenol-formaldehyde resins were diagnosed when routinely patch tested with P-F-R-2 in addition to PTBP-F-R. Although patch testing with a mixture of both resins was not as good as patch testing with the 2 resins separately, it was better than testing only with PTBP-F-R, since 1.6 times more patients with contact allergy to phenol-formaldehyde resins were still diagnosed. P-F-R-2 is therefore recommended for routine patch testing, preferably as a separate patch test but otherwise as a mixture with PTBP-F-R. PMID- 2972521 TI - Silver polish: another source of contact dermatitis reactions to thiourea. AB - A cleaning woman in a restaurant became sensitized to thiourea in a silver polish. The dermatitis persisted for several months and occasionally flared up after exposure to sunlight, even after she no longer used the product. Patch testing and photopatch testing showed that she had both contact and photocontact allergy. PMID- 2972522 TI - Dermatitis from crystal violet. PMID- 2972523 TI - The release of nickel from blackboard chalk may cause contact dermatitis. PMID- 2972524 TI - Contact dermatitis due to Ziram and Maneb. PMID- 2972525 TI - Protein contact dermatitis in food handlers. PMID- 2972526 TI - Allergy to subcutaneous heparin. PMID- 2972527 TI - Contact reaction to flame retardant. PMID- 2972528 TI - Chromium dermatitis caused by epoxy resin. PMID- 2972529 TI - Contact dermatitis from azodicarbonamide in earplugs. PMID- 2972531 TI - Determinants of atrial natriuretic factor in the adult respiratory distress syndrome. AB - We measured plasma atrial natriuretic factor over the clinical course of 12 patients with the adult respiratory distress syndrome (ARDS). A total of 33 sequential measurements were correlated with 23 hemodynamic, pulmonary, and renal parameters. Atrial natriuretic factor was found to be eight times higher than age matched healthy control subjects (p = 0.001). Although atrial natriuretic factor correlated well with pulmonary capillary wedge pressure (r = 0.592), there was no significant relationship between right atrial pressure and plasma atrial natriuretic factor (r = 0.258). Atrial natriuretic factor was best related to pulmonary artery systolic pressure (r = 0.751). We conclude that in ARDS, plasma atrial natriuretic factor is markedly elevated and arterial levels are best determined by left atrial pressure and pulmonary hemodynamics, and not by right sided cardiac pressures. The role of increased circulating atrial natriuretic factor in the pathophysiology of ARDS is to be clarified in future studies. PMID- 2972530 TI - Monoclonal antibodies to heparan sulfate proteoglycan: development and application to the study of normal tissue and pathologic human kidney biopsies. AB - Monoclonal antibodies (mAbs) (4F2 and 7E12) were prepared against heparan sulfate proteoglycan (HSPG) isolated from bovine glomeruli. Enzyme linked immunosorbent assays (ELISA) and immunoblotting demonstrated that the mABs reacted with HSPG. Indirect immunofluorescence (IF) showed that the mAbs stained renal basement membranes (BMs) and BMs in other organs of normal bovine and human tissues in patterns typical of HSPG. Immunoinhibition studies, and immunoblotting of heparan lyase digested HSPG, indicated that the mAbs recognize HSPG core protein. In kidney biopsies from patients with acute poststreptococcal GN, intact linear glomerular BM (GBM) staining for HSPG was noted despite markedly widened capillary loops. In membranous and in diffuse proliferative lupus GN, loss of HSPG staining was demonstrated at sites of immunodeposition of IgG or C3, while increased staining for HSPG was noted in areas of newly formed GBM. Extensive loss of HSPG was seen in areas of glomerular sclerosis and necrosis. In biopsies from patients with minimal change glomerulonephritis (GN) and mesangioproliferative lupus GN, a normal linear GBM distribution of HSPG was noted. The findings are discussed in the context of current knowledge regarding the pathogenesis of glomerular injury. MAbs to BM HSPG should prove useful for future immunochemical studies, and for the study of diseases of the basement membrane. PMID- 2972532 TI - Familial sudden death. Report of a case and review of the literature. AB - A 15 1/2-year-old boy died suddenly while swimming. He had a family history of sudden death involving three consecutive generations, including a brother. The patient had a history of exercise-related syncope, for which he was being treated with nadolol. Autopsy showed enlarged heart, normal coronary arteries, right ventricular septal hypertrophy, quadricuspid pulmonary valve, accessory tricuspid valve, and a moderately elongated and thickened mitral valve. Conduction system revealed that the penetrating bundle was pushed to the left side of the summit of the ventricular septum by the right ventricular septal hypertrophy; it was lobulated and showed fatty-fibrous changes. These findings extended throughout the beginning of the bundle branches. We conclude that in this patient with familial sudden death and normal QT interval, the abnormal right ventricular septal hypertrophy altered the course and produced degenerative changes in the conduction system, which may have caused sudden death. PMID- 2972533 TI - [Opening of chronic coronary artery occlusions with a recanalization catheter]. AB - Percutaneous transluminal angioplasty for recanalization was attempted for 44 chronic coronary arterial occlusions in 41 patients (two occluded vessels in three patients). In 11 instances the occlusion could be passed with a guide-wire. In the other 33 this was not possible. In 25 of them a 3, 4 or 5 F recanalization catheter, its end tapered to 2 or 3 F, was used. In this way re-opening was possible in 17 cases (68%). The catheter made it possible to splint or stiffen the guidewire to keep it straight, superselectively inject contrast medium, measure the pressure distal to the occlusion, and gradually bougie-like enlarge the resulting subtotal stenosis. A total of 28 of 44 coronary occlusions were re opened (64%). All these patients were functionally improved. At subsequent angiography (a mean of 3.6 months later) 22 of 25 vessels had remained open, but ten had narrowed and three had become re-occluded. PMID- 2972534 TI - Influence of a high-fat ration on parameters of defence against infection in weaned pigs. PMID- 2972535 TI - Review article: immune nephritides due to malaria. PMID- 2972536 TI - [Inhibiting effect of a chalone-containing extract of calf pancreatic tissue on the proliferative activity of normal and tumorous pancreatic cells in vitro and in vivo]. AB - The pancreatic chalone has been studied for its effect on the proliferative activity and morphology of normal and tumour cells of the pancreas. These findings in experiments in vitro were confirmed by the data of the in vivo experiments. The results show that pancreatic chalone inhibits cell proliferation, decreases mitotic index and causes morphologic changes in cultivated pancreatic cells. PMID- 2972537 TI - Laparoscopic findings in serosal eosinophilic gastroenteritis. Report of two cases. AB - This report describes the laparoscopic features of two patients with serosal eosinophilic gastroenteritis. In one case only hyperemia of the peritoneal serosa was found. In the other the laparoscopic picture resembled peritoneal carcinomatosis. We show that laparoscopy and peritoneal biopsy may permit diagnosis of the disease without the need for laparotomy. PMID- 2972539 TI - PL of coliphage lambda: an alternative solution for an efficient promoter. AB - Promoter PL of coliphage lambda is highly active in vivo although it is recognized 15-30 times less efficiently by RNA polymerase when compared with promoters of similar strength. Moreover, it differs significantly from the consensus sequence for Escherichia coli promoters. Sequence variants of PL which are more homologous to consensus promoters bind RNA polymerase with increased efficiency. They are nevertheless significantly reduced in their in vivo strength. High activity can be restored by a downstream sequence of a typical consensus-like promoter. Evidently, such elements are required for the efficient release of a stably bound RNA polymerase into a transcriptional elongation complex. We propose that the functional programme encoded in a promoter sequence can be optimized in alternative ways. PMID- 2972538 TI - Rapid turnover of adenovirus E1A is determined through a co-translational mechanism that requires an aminoterminal domain. AB - The product of the adenovirus E1A 13S mRNA can both stimulate and repress the expression of certain viral and cellular genes. As with several other regulatory proteins, E1A has a short half-life, approximately 40 min. Although this short half-life is observed in cells expressing the E1A gene, it is not the case with cells injected with E1A protein, where its half-life is very long, generally greater than 15 h. We have sought to reconcile these apparent differences in E1A stability. Using Xenopus oocytes, we find that E1A exhibits its characteristic short half-life when it is synthesized from injected mRNA while it has a very long half-life when it is injected as a protein synthesized originally in Escherichia coli or reticulocyte lysates. In order to delineate the amino acids responsible for rapid E1A turnover, several deletion mRNAs were constructed, injected into oocytes, and E1A half-life determined. Carboxyl-terminal deletions and an internal deletion of residues 38-86 failed to increase the half-life of E1A. In contrast, amino-terminal deletions of 70 and 14 residues resulted in very stable E1A proteins (t1/2 greater than 20 h). Furthermore, deletion of the second amino acid, an arginine, resulted in a stable E1A protein. The amino-terminal region of E1A was able to induce the rapid turnover of a normally stable protein, beta-globin, in oocytes injected with an E1A-globin chimeric mRNA. This E1A induced instability of globin was abolished, however, when the protein was first synthesized in reticulocyte lysates and then injected into oocytes. The amino terminal region of E1A is also important in governing halflife in adenovirus infected HeLa cells. These results demonstrate that the half-life of E1A is established cotranslationally through a mechanism involving sequences within the amino-terminal 37 residues. PMID- 2972541 TI - Molecular cloning of murine p68, a Ca2+-binding protein of the lipocortin family. AB - A plasmid cDNA library prepared from a T-lymphocyte clone of murine strain B10.A origin was screened by cross-species DNA hybridisation using a partial human p68 cDNA clone, identified as containing coding sequences for previously determined amino-acid sequences. The longest p68 cDNA insert from this library and a full length cDNA insert from a second similar library were fully sequenced. A comparison of the derived amino-acid sequence with that of human p68 revealed extensive homology (95% overall). Homology at the nucleotide level was 89% in the open reading frame and 85% and 50% in the 5' (33 nucleotides) and 3' (347 nucleotides) non-coding regions respectively. Eight segments of internal homology were observed, each containing a highly conserved consensus region of 17 amino acids correlating with that described for several membrane associated calcium binding proteins [Geisow, M. J., Fritsche, U., Hexham, J. M. & Johnson, T. (1986) Nature (Lond.) 320, 636-638]. These results provide further evidence that p68 is a member of the same gene family as p32,p36 and lipocortin I and demonstrate an unusually high level of inter-species sequence conservation of p68 between mouse and human. PMID- 2972540 TI - Epidural sufentanil for post-operative pain relief: effects of adrenaline. AB - The analgesic, respiratory and haemodynamic effects of epidural sufentanil 75 micrograms (Group 1) or sufentanil 75 micrograms with adrenaline 75 micrograms (Group 2) were studied in 20 patients following abdominal surgery in a double blind randomized trial. Pain relief, assessed on a linear analogue scale, sedation, heart rate (HR), mean arterial pressure (MAP), respiratory rate (RR) and arterial carbon dioxide tension (PaCO2) were recorded before, and for 12 h after, injection. Good post-operative pain relief was obtained after 4 min in Group 1 and 6 min in Group 2. All patients in both groups were pain-free after 30 min. Analgesia lasted for 450 +/- 46 min in Group 1 and 690 +/- 92 min in Group 2 (P less than 0.05). Coughing and active movement was possible with little pain for 6 h post-operatively in Group 1 and 7 h in Group 2. The respiratory rate was significantly decreased from 15 min to 1 h after injection in Group 1 compared with Group 2 (P less than 0.05) and in three patients in Group 1 it was necessary to stimulate the patients verbally to maintain adequate respiration. PaCO2 increased significantly in both groups over the first 2 h. The patients in Group 1 showed more marked sedation 1 h after injection. Changes of HR and MAP were similar in both treatment groups. Side-effects were observed more frequently in Group 2, although the differences were not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972543 TI - Electrocardiographic alterations suggestive of myocardial injury elicited by rapid pressure lowering in hypertension. AB - In hypertensive cardiac hypertrophy, the elevated coronary perfusion pressure compensates importantly for the raised coronary resistance. An imbalance between perfusion and left ventricular (LV) mass, such as that occurring with rapid or excessive blood pressure lowering, may result in an inadequate oxygen supply. In 28 primary hypertensives (Group A) with LV mass index within the mean + 1 SD (96 + 19 g m-2) of 145 controls, and in 26 patients whose LV mass exceeded these values (Group B), we lowered the diastolic blood pressure rapidly to 85-90 mmHg, using both s.l. nifedipine and i.v. nitroprusside. During each test, eight patients in Group B had inversion of T waves in lead I, aVL, V3-V6, which waxed and waned in parallel with the pressure fall and recovery, and was independent of conduction disturbances, variations or group differences in the QRS axis, QTc interval, heart rate, LV fractional shortening and wall stress. A 'coronary steal phenomenon' or passive collapse in compliant lesions consequent to vasodilatation may trigger acute myocardial ischaemia in the presence of severe coronary disease. Patients developing the ECG alterations, however, were free from angina and four, who were subjected to coronary angiography, had normal arteriograms. Patients with the myocardial injury pattern showed greater LV mass indices and larger falls in diastolic pressure for it to reach normal levels. The supply of energy to the hypertrophied hypertensive heart seems to depend importantly on the coronary perfusion pressure, suggesting the cautious use of rapid acting drugs. PMID- 2972542 TI - Improvement of myocardial perfusion after thrombolysis assessed by thallium-201 exercise scintigraphy. Report of the Working Group on Thrombolytic Therapy in Acute Myocardial Infarction of the Netherlands Interuniversity Institute of Cardiology. AB - The effects of recanalization by early intracoronary streptokinase in acute myocardial infarction on myocardial perfusion were studied in a subset of 236 out of 533 patients enrolled in a prospective multicentre trial, randomly allocated to either conventional treatment or to thrombolysis. Maximal symptom-limited thallium-201 stress testing was performed nine to 14 (median 12) weeks after the acute event in 236 patients with a first myocardial infarction, of whom 108 patients were allocated to conventional treatment and 128 patients to thrombolysis. Three-view thallium exercise and redistribution scintigrams were divided into eight segments. The number of normal thallium segments at redistribution was 5.3 +/- 1.6 (mean +/- SD) in the control group and 5.8 +/- 1.6 in the thrombolysis group (P less than 0.01), indicating increased myocardial perfusion after early thrombolytic treatment. This beneficial effect was particularly evident in patients with a first anterior infarction with 4.8 +/- 1.6 normal thallium segments in the control group and 5.9 +/- 1.6 normal segments in the thrombolysis group (P less than 0.001). There was no difference in residual ischaemia during exercise between the control and the thrombolysis group either in the patients with anterior or in those with inferior infarction. It is concluded that early thrombolysis within 4 h after onset of symptoms, in some of the patients followed by angioplasty or coronary artery bypass surgery, leads to sustained improvement of myocardial perfusion at rest without excessive ischaemia during exercise, particularly in patients with acute anterior infarction. PMID- 2972544 TI - Platelet activation in angina at rest. Evidence by paired measurement of plasma beta-thromboglobulin and platelet factor 4. AB - Indexes of in vivo platelet activation, beta-thromboglobulin and platelet factor 4 were measured in triplicate in plasma from venous blood of 69 patients with proven ischaemic heart disease (IHD), discarding samples with a ratio of the plasma concentrations of the two proteins less than 2.6, in order to rule out sampling artifacts. Compared with 60 control volunteers, differences were not significant [for beta-thromboglobulin controls (ng ml-1, mean +/- SD) 27.8-8.6, ischaemic patients 32.3 +/- 17.1; for platelet factor 4 controls 4.3 +/- 1.4, ischaemic patients 5.9 +/- 5.7]. However, when patients were stratified according to disease activity (Group I--patients without spontaneous ischaemic episodes at rest during 4 days of continuous electrocardiographic monitoring; Group II- patients with less than 1 ischaemic episode/day; Group III--patients with greater than 1 episode/day), these indexes were increased in 'active' patients (for beta thromboglobulin, in Group II--32.4 +/- 10.5 ng ml-1, P less than 0.05 vs. Group I; in Group III--42.6 +/- 14.6 ng ml-1, P less than 0.01 vs. Group I, P less than 0.05 vs. control. Platelet factor 4 was increased only in Group III--8.9 +/- 7.2 ng ml-1, P less than 0.05 vs. control). Beta-thromboglobulin and platelet factor 4 were 25.0 +/- 6.7 ng ml-1 and 4.9 +/- 4.8 ng ml-1, respectively, in Group I (P = NS vs. control). A relationship with the number of spontaneous ischaemic episodes at rest was confirmed by linear regression analysis (in Group III patients for beta-thromboglobulin: r = 0.76, P less than 0.01, and for platelet factor 4 r = 0.62, P less than 0.01). Levels were not elevated in patients with previous myocardial infarction without ischaemia at rest and/or patients with stable angina, and were not influenced by the occurrence of a positive exercise stress test. Coronary angiograms of ischaemic patients were analyzed to assess the extent and severity of atherosclerotic involvement: for both extent and severity, involvement was similar in the three groups. These data support the hypothesis of the occurrence of platelet activation in patients with spontaneous angina at rest, but not in other subsets of IHD patients, and establish the possibility of detecting in vivo platelet activation in IHD by means of such circulating markers. PMID- 2972545 TI - The efficacy of GR38032F, an antagonist of 5-hydroxytryptamine-3 (5-HT3) in the prophylaxis of cisplatin (CDDP)-induced nausea and vomiting. PMID- 2972546 TI - Presence and characterization of insulin-like growth factor 1 receptors in human benign breast disease. AB - Insulin-like growth factor 1 binding sites were characterized in human benign breast disease. We demonstrated the presence of one high affinity binding site. Chemical cross-linking of [125I]IGF1 to benign breast disease membranes in reducing condition and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed one band with an apparent Mr of 130,000. The specificity of the binding was studied: IGF 2 was a good competitor whereas insulin competed for binding with a potency lower than 1/100 that of IGF1. This IGF1 binding corresponded to the previously described type I IGF receptor (IGF1-R). IGF1-R was assayed in 35 cases of benign breast disease and two samples of normal breast tissue. Forty-three per cent of the lesions were IGF1-R positive. The mean geometric level of specific binding was 1.98% in the whole population, it was significantly lower in adenofibromas (1.55%) than in epithelial hyperplasia (2.5%); it was 2% in dystrophic disease. IGF1-R was undetectable in normal tissue. Considering our previous results showing that almost all the breast cancers contained IGF1-R, these data suggest that the increase in IGF1-R could be a marker of malignant tumor development. PMID- 2972547 TI - Immunological characterization and clinical significance of low mobility cells appearing in the peripheral blood mononuclear cells of cancer patients. AB - The characteristics of low mobility cells (LMC) in the peripheral blood mononuclear cells (PBMC) of cancer patients were studied using a Parmoquant-L, a fully automated electrophoresis instrument, and a fluorescence-activated cell sorter (FACS). The L/H ratio (%) of LMC (less than 0.95 micron/s/V/cm) to high mobility cells (HMC) (greater than or equal to 0.95) was used to analyze the electrophoretic mobility (EPM) histogram of PBMC. In the cancer patient, the L/H ratio increased as a result of the appearance of LMC. As the ratio closely correlated with clinical stage of cancer, recurrence of cancer, and performance status in the patients, the method is a useful parameter for prognosis of cancer, practically, for early detection of recurrence in the follow-up study of cancer patients. The EPM of monocytes and subsets of lymphocytes was determined to characterize LMC. Monocytes were in LMC. T-Cells such as helper/inducer (Leu-3+), suppressor (Leu-2+Leu-15+) and cytotoxic (Leu-2+Leu-15-) cells and natural killer (NK) cells (Leu-7+ or Leu-11+) were in HMC. The percentage of monocytes correlated significantly with the mean EPM of PBMC. In cancer patients, the percentage of monocytes increased significantly. On the other hand, suppressor T cells did not increase enough to account for LMC. The appearance of suppressor T cells correlated with that of monocytes. These results suggest that LMC in cancer patients consisted of mainly monocytes rather than suppressor T-cells. PMID- 2972548 TI - The influence of antihistamines on human performance. AB - We have studied the effects of three antihistamines on task performance in two separate experiments. Healthy subjects were tested at weekly intervals in a double-blind, Latin square design. In Experiment I the subjects were treated orally with loratadine 10 mg, clemastine 1 mg, terfenadine 60 mg, or placebo. In Experiment II 5 mg diazepam was given orally with each of the four treatments used in Experiment I. In both experiments subjects' performance was evaluated in reaction time and tracking tasks after treatment. In both experiments, the tracking task initially was performed alone and then simultaneously with a continuous memory task; the subject also graded their mental status on visual analogue rating scales. In both experiments task performance was not generally impaired after treatment with loratadine or terfenadine. The concomitant administration of diazepam in Experiment II appeared not to affect subjects' performance. However, clemastine caused a decay in subjects' performance in both Experiments I and II, but only on the tracking task. At the conclusion of both experiments, sleepiness was reported by more subjects when treated with clemastine than when treated with loratadine, terfenadine, or placebo. PMID- 2972549 TI - Attenuation of cutaneous reactivity to histamine by cetirizine and dexchlorpheniramine. AB - A double-blind cross-over study was performed in 12 healthy female volunteers comparing cetirizine di-HCl (10 mg) and sustained release dexchlorpheniramine maleate (6 mg) with respect to attentuation of histamine-induced skin wheals and subjective central nervous system (CNS) effects. Cetirizine was significantly more effective than dexchlorpheniramine in suppressing the size of wheals from 2 to 24 h after drug administration. In fact, at 24 h cetirizine was still as effective as 2 h after ingestion. Ten subjects receiving dexchlorpheniramine reported subjective symptoms relating to CNS depression, in contrast to only one subject given cetirizine. PMID- 2972551 TI - Characterisation of receptors mediating the contractile effects of prostanoids in guinea-pig and human airways. AB - Receptors mediating the contractile effects of prostanoids in the airways of guinea-pig and man have been compared using a range of selective agonists and antagonists. In guinea-pig and human lung parenchymal strips, contractile responses to all prostanoid agonists were blocked by EP092 and AH23848 (antagonists of thromboxane-sensitive receptors) but were unaffected by the EP1 receptor antagonist, AH6809. In guinea-pig tracheal rings responses to the thromboxane mimetic U46619 were antagonised by EP092 and AH23848. AH6809 was ineffective, but did antagonise responses to 16,16-dimethylPGE2 and PGF2 alpha in this tissue. In the human bronchiolar ring responses to U46619 were poorly antagonised by AH23848, EP092 and AH6809. The results suggest that prostanoid contractile agonists act through thromboxane receptors in guinea-pig and human lung parenchymal strips and through thromboxane and EP1 receptors in guinea-pig trachea. A novel prostanoid receptor type, or subtype, mediating contractile effects may be present in human bronchioles. PMID- 2972550 TI - Inhibition of the aldosterone response to sodium depletion in man by stimulation of dopamine DA2 receptors. AB - In this study we have investigated the effect of co-dergocrine, a selective DA2 agonist, on plasma aldosterone concentrations (PAC) in twelve patients with essential hypertension both in basal conditions and during sodium depletion. Sodium depletion resulted in an increase of PAC from 38 (13) pg/ml to 297 (21) pg/ml. The PAC response to sodium depletion was reduced to 155 (29) pg/ml by co dergocrine. No significant PAC changes were found in patients maintained on a normal sodium intake. In addition the drug did not significantly modify plasma renin activity (PRA) in either experimental group. These results suggest that the dopaminergic inhibition of aldosterone secretion in man is mediated by DA2 receptors in the adrenal cortex. PMID- 2972552 TI - 5-Hydroxytryptamine1A receptors are linked to a Gi-adenylate cyclase complex in rat hippocampus. AB - The ability of 5-hydroxytryptamine (5-HT) and the 5-HT1A selective agonist 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) to modulate adenylate cyclase activity was measured in rat hippocampus. In vitro ADP ribosylation of GTP binding proteins by pertussis toxin in this tissue abolished both 5-HT- and 8-OH DPAT-induced inhibition of forskolin-stimulated adenylate cyclase activity. These findings indicate that 5-HT1A receptors are linked a pertussis-sensitive Gi protein in rat hippocampus. PMID- 2972553 TI - Cephalometric comparison between children with and without Down's syndrome. PMID- 2972554 TI - Thermal sensitivity of Clostridium botulinum type C toxin. AB - A sterile suspension containing 950 mouse LD50 per ml of type C botulinum toxin was exposed for various periods to different temperatures. The time required for the 99% (hundred-fold) reduction of toxicity was more than 5 years at -70 degrees C or -20 degrees C, 6 months at +5 degrees C, 3 weeks at +20 degrees C, 2 weeks at +28 degrees C, 2 days at +37 degrees C, 9 h at +42 degrees C, less than 30 min at +56 degrees C, less than 20 min at +60 degrees C, and below 5 min at +80 degrees C. The results suggest that Clostridium botulinum type C toxin, if produced in an ecosystem of the mild climatic zone, might persist there over the winter season and cause the intoxication of vertebrates next early spring in the absence of further microbial toxigenesis. PMID- 2972555 TI - Fc receptor-mediated binding and ingestion of immunoglobulin G-coated erythrocytes by the epithelium of the posterior ciliary processes: an in-vitro study. AB - In the present in vitro study of intact ciliary processes, we observed, under differential interference microscopy, IgG inhibitable selective epithelial adherence and internalization of IgG-coated erythrocytes (E-IgG). Unequivocal evidence of ligand ingestion was obtained with a fluorescence technique which employed glutaraldehyde-fixed opsonized erythrocytes. Neither adherence nor internalization were seen when uncoated erythrocytes, IgM, or F (ab')2-coated erythrocytes were substituted for E-IgG in the incubation media. These in-vitro studies demonstrated that the cell membranes of this epithelium exhibit a specific receptor for IgG which may clear the aqueous of IgG aggregates and be associated with the exocytosis of soluble mediators of inflammation. PMID- 2972556 TI - IGF-I receptors in the bovine neural retina: structure, kinase activity and comparison with retinal insulin receptors. AB - The retina contains specific high-affinity receptors for insulin-like growth factor-I (IGF-I). Although IGF-I binding was observed in photoreceptor outer segments, the level of this binding was only 10% of that found in whole retina or mixed preparations of rod outer (ROS) and inner (RIS) segments. The higher IGF-I binding activity in RIS and non-photoreceptor regions of the retina suggests these sites as candidates for putative IGF-I action. Data from crosslinking experiments with and without neuraminidase treatment indicate that the binding subunits of the retinal IGF-I receptor exist in two subpopulations (Mr = 121- and 131 kDa), and that the larger of the two subunits has either a greater number or more exposed sialic acid residues. In these characteristics, the retinal IGF-I receptor is similar to the retinal insulin receptor. Retinal IGF-I and insulin receptors possess kinase activity towards their own beta-subunits, a tyrosine containing copolymer, and various molecular forms and subunits of transducin (T alpha-GDP, T alpha-GTP, T beta). The transducin forms are phosphorylated with different efficiencies (e.g. T alpha-GDP is 10-15 times more effective than T alpha-GTP as substrate). These differences are also observed in basal conditions and may reflect differences in transducin subunit affinity for the IGF-I and insulin receptor. In all retinal areas examined, tracer IGF-I binding is 10 to 20 fold higher than insulin binding; however, autophosphorylation levels are approximately equal. PMID- 2972558 TI - Retinoic acid regulates IgG Fc receptor expression in human myelomonocytic leukemia cells and normal peripheral monocytes. AB - The regulation of IgG Fc receptor (Fc gamma R) expression by retinoic acid (RA) in human myelomonocytic cells at different stages of maturation was studied. RA suppressed IgG-coated erythrocyte (EA) rosette formation of myelomonocytic cells blocked at relatively late stages of differentiation such as ML-1, U-937, THP-1 T, normal monocytes, and fresh cells of patients with acute myelomonocytic leukemia. However, RA increased the percentage of EA rosetting promyelocytes of HL-60 and of patients with acute promyelocytic leukemia and a part of myeloblasts isolated from acute myelogenous leukemia patients. Other myeloblasts including KG 1a, KG-1, and fresh cells from patients with acute myelogenous leukemia were not affected. A kinetic study using HL-60 and THP-1-T demonstrated that an increase required at least a 48-h exposure and that the maximum decrease required approximately 6 h. The RA effect on both cell lines was dose-dependent. The number of Fc gamma R of HL-60 and THP-1-T treated with RA became very close, although untreated THP-1 had almost 10 times as many as HL-60. Kd for IgG in both THP-1-T and HL-60, either untreated or treated with RA, remained unchanged. These observations indicate that one of the important roles of RA is regulation of Fc gamma R expression in myeloid cells. PMID- 2972557 TI - The prevalence of serological markers, hepatitis B virus DNA and elevated serum amino transferase values in institutionalized mentally handicapped males and females: an epidemiological study of HBV infection in residents with Down's syndrome or other forms of mental deficiency. AB - Conventional HBV serological markers, as well as serum alanine amino transferase and hepatitis B virus DNA (HBV DNA) were studied in a population of 667 institutionalized mentally handicapped males and females and in 676 staff members. The role of Down's syndrome (DS), sex and age-related factors with respect to the prevalence of these markers was analyzed. A young age at admission was found as one of the important factors in the development of the chronic HBsAg carrier state in females. The well-known higher prevalence of HBsAg carriership in DS patients appeared to be restricted to males. Markers indicative of viral replication, HBeAg and HBV DNA, were also more prevalent in male than in female DS residents, or OMR residents. These findings indicate that the phase of active viral replication is prolonged in male HBsAg carriers with DS. However, only 66.7 per cent of HBV DNA-positive and 26.2 per cent of HBsAg-positive residents had abnormal serum alanine aminotransferase (ALT) values. In addition, more than 50 per cent of residents with elevated serum alanine amino transferase values were negative for HBV serological markers. We conclude that male DS residents are the main source of HBV infection in institutions for the mentally handicapped and that determination of ALT values is not very useful in identifying those HBsAg carriers capable of transmission the infection. PMID- 2972559 TI - An incremental method to assess the linearity of gas flowmeters: application to Fleisch pneumotachographs. AB - A new method of studying the linearity of gas flowmeters was tested on different models of Fleisch pneumotachographs. The method applies a steady flow to the test flowmeter, which is increased in a stepwise manner by adding a constant flow increment. This is achieved using two flow sources in parallel. The method does not require any reference flow channel and may be implemented with standard laboratory equipment. Using this method, the gain of Fleisch pneumotachographs, whatever their size, decreased by about 2-3% from low flows to about 40% of their nominal full scale (FS), and then increased almost linearly with increasing flow. The error was 8-13% at 200% FS. The following equation was devised to correct the data at high flow: Vc = Vt (1-K (Vt-S] where Vc and Vt are the corrected and measured flow respectively, K a gain correction factor and S a flow threshold below which no correction is needed. Applying this correction with suitable coefficients, the maximal error was below 3% from 0-200% FS. PMID- 2972560 TI - [Potentiometry applied to the characterization and analysis of polysaccharides for pharmaceutical use]. PMID- 2972561 TI - Estrogen-controlled gene expression in tissue culture cells by zearalenone. AB - In two estrogen-sensitive cell lines, Le42 and MCF-7, the estrogenic potential of the nonsteroidal mycotoxin zearalenone has been investigated. The chloramphenicol acetyltransferase (CAT) gene expression in Le42 cells is induced by zearalenone after transfection with a CAT-gene construct controlled by an estrogen responsive element [(1986) Cell 46, 1053-1061]. In MCF-7 cells zearalenone induces at least 2 exoproteins (52 and 160 kDa) which are estrogen-specific [(1980) Cell 20, 353 362). These data suggest that zearalenone acts by activating the estrogen receptor. Due to the high sensitivity of these cell lines for zearalenone both test systems are proposed as assays for a quantitative estimation of the biological (estrogenic) activity of this widespread mycotoxin. PMID- 2972563 TI - Acute surgical intervention for complications of percutaneous transluminal angioplasty. AB - Since 1979, percutaneous transluminal angioplasty (PTA) resulting in an overall improvement of 80.3% has been performed in 4380 patients with occlusive atheroma of the lower extremities. Complications requiring immediate surgical treatment occurred in 123 (2.8%) of the cases, consisting of occlusion (16%), dissection (13%), perforation (10%), embolisation (15%) and haematoma at the puncture track (69%). Of the group treated surgically, 18 (14.6%) patients underwent amputation of a lower limb. Since 1980 348 percutaneous transluminal coronary angioplasties (PTCAs) have been performed for coronary artery disease. In 15 cases (4.3%) emergency coronary bypass surgery was necessary because of complications encountered with PTCA. Two patients died and perioperative myocardial infarction occurred in 7 (subendocardial-3: transmural-4). In spite of a low complication rate and good results from PTA and PCTA acute surgical intervention for complications carries a significant operative morbidity and mortality. We conclude that selection for PTA and PTCA has to involve vascular and cardiac surgeons, their presence being mandatory at institutions in which therapeutic radiological procedures are undertaken. PMID- 2972562 TI - The role of balloon angioplasty in the management of lower limb ischaemia. AB - Three hundred and twenty-three angioplasties (159 iliac; 164 femoro-popliteal) were performed for lower limb ischaemia on 253 consecutive patients from 1980-86. The mean resting ankle/brachial pressure index (ABPI) was increased at one month in open angioplasties as follows: Femoro-popliteal 0.56-0.82; iliac with open superficial femoral artery (SFA) 0.7-0.95; iliac with occluded SFA 0.52-0.63. Successful angioplasty virtually abolished the post-exercise fall in ABPI except for patients having iliac dilations when multisegment disease was present. At 5 years, cumulative patency was 72% for iliac angioplasty and 53% for femoro popliteal angioplasty. The results of angioplasty were compared with operative arterial reconstructions during 1985. Eighty-eight (91%) of 96 surgical bypasses were alive and patent at hospital discharge and 44 (71%) of 62 angioplasties were patent at 1 month. There were many fewer complications with angioplasty which involved a much shorter hospital stay (2 days vs 16 days). PMID- 2972564 TI - [Experience with health education work]. PMID- 2972565 TI - Laparoscopic CO2 laser vaporization of endometriosis compared with traditional treatments. AB - This study reports on 108 infertile patients with endometriosis diagnosed at laparoscopy. Sixty-four patients had endometriosis and adhesions vaporized with a CO2 laser (LAS) and were prospectively compared with a control group (CON) of 44 patients who had laparoscopy. The 6- and 12-month estimated cumulative pregnancy rates for LAS were 0.32 +/- 0.07 and 0.55 +/- 0.09 respectively, and for CON 0.17 +/- 0.06 and 0.43 +/- 0.09 (Breslow P = 0.10). Monthly fecundity rates were 6.7% in LAS and 4.5% in CON. Survival analysis with fixed covariates showed that pregnancy rates were increased in patients with adhesions (P = 0.002) and other pelvic disease (P = 0.0001). Pregnancy rates were reduced by age (P = 0.02), previous adhesiolysis (P = 0.0000) and post-laparoscopy medical treatment (P = 0.0002). Our findings indicate that CO2 laser laparoscopy vaporization of endometriosis can be a safe, effective, and possibly improved modality for treating endometriosis. PMID- 2972566 TI - Atrial natriuretic peptide, plasma renin activity, and aldosterone in women on estrogen therapy and with premenstrual syndrome. AB - Estrogens are known to increase the renin-angiotensin-aldosterone system and to produce fluid retention, while atrial natriuretic peptide (ANP) induces an increase of the urinary output and tends to return the fluid balance to normal. The aim of this study was to test whether the levels of ANP were decreased during chronic estrogen and progestin administration, thereby possibly decreasing the amount of fluid excreted. The authors also studied women with premenstrual syndrome (PMS), because of the associated fluid retention often described with this syndrome. Levels of ANP, plasma renin activity (PRA), and aldosterone were determined in premenopausal women in the early follicular phase (EFP) and on low dose oral contraceptives (OC), in postmenopausal patients with and without estrogen replacement therapy (ERT), and in women with PMS associated with fluid retention. The concentrations of ANP and PRA were enhanced in the women on OC, but those of aldosterone were unchanged. No differences were observed in the women on ERT or with PMS. It is concluded that the levels of PRA and ANP are affected by estrogen or progesterone therapy or the combination of the two and this response is dose dependent or additive. Furthermore, ANP and PRA do not seem to play a direct role in PMS. PMID- 2972567 TI - [Functiono-structural aspects of the adaptation of the heart to physical loads]. PMID- 2972568 TI - [250 years of the teaching of physiology at the Petersburg-Leningrad University]. PMID- 2972569 TI - Treatment of the geriatric dentally phobic patient. AB - The elderly patient, who is phobic or exceedingly anxious established behavior patterns years ago when dentistry was not as advanced as today and when dental experiences were unpleasant and uncomfortable. The elderly phobic patient may view his role as a passive participant as he was in his youth; that is, he fears that the dentist will treat him as he was treated by an unsympathetic dentist years ago. In order to create acceptable, positive experiences for the phobic or anxious patient, the first appointment should give the patient an opportunity to know the dentist and office staff. The dentist can take this opportunity to reassure the patient that pain does not have to be a part of dental treatment. It is imperative that the dentist and staff involve the patient and instill trust. One must always have the patient's perspective in mind and constantly, by word or deed, assure the patient that he will always be in control of decisions and treatment. The dentist must not view the geriatric patient as different from his other patients. He or she must be perceptive and deliberate in the initial encounter and early relationship and determine what is the best treatment for that patient. The dentist must be realistic about the goals and requirements of that patient, for whom the dentist's ultimate objective should be to achieve quality dental care, done expeditiously, without fear and pain. PMID- 2972570 TI - Treatment of the agoraphobic dental patient. AB - The dental-phobic patient is rarely motivated to seek treatment. When this type of patient does, it is difficult to perform the dental work without repeated interruptions due to the high anxiety being displayed. Many dental phobics are actually agoraphobic, and this further complicates the matter. The agoraphobic dental patient mislabels any physiologic changes that are experienced as a prelude to disaster, or horrific consequences, and something there is no control over. This type of patient looks to depend on "safe" places (like home) and "safe" people (like spouses and doctors). The physiologic changes that are perceived may be induced pharmacologically (for example, numbness and tingling from the lidocaine, imbalanced blood gases due to hyperventilation), physically (for example, tilt of the dental chair, pressure in the mouth), or cognitively (autonomic excitation due to negative thoughts). The inability to mitigate the physiologic changes or symptoms confirms the patient's irrational belief that disaster is inevitable. Two basic considerations must be given priority when treating agoraphobic dental patients. These are: (1) providing them with a sense of being in control of the situation and/or their symptoms, and (2) training them to relabel or reinterpret their symptoms objectively as nervousness, or some simple and explainable physiologic change that should be present. Therefore, teaching agoraphobic dental patients cognitive-behavioral procedures such as relaxation and cognitive-restructuring techniques works quite well. Both will help to prevent, or reduce to a manageable level, the symptoms, and result in successful dental work. PMID- 2972571 TI - Integrating the dental phobic into the general dental practice. PMID- 2972572 TI - Eruptive pyogenic granulomas and acne fulminans in two siblings treated with isotretinoin. A possible common pathogenesis. AB - We report the case of two siblings, one of whom presented isotretinoin-induced eruptive pyogenic granulomas in the course of severe acne, and the other developed isotretinoin-induced acne fulminans. A possible underlying common pathogenic process of these two side-effects of isotretinoin is discussed. PMID- 2972573 TI - Acetylsalicylic acid may induce a lichenoid eruption. PMID- 2972574 TI - Dentists and AIDS. PMID- 2972575 TI - Alterations in T-lymphocyte subpopulations in type I diabetes. Exploration of genetic influence in identical twins. AB - To evaluate factors influencing the alteration in subsets of T-lymphocytes, we studied 24 pairs of identical twins discordant for insulin-dependent (type I) diabetes mellitus. Subsets were assessed by monoclonal antibodies and a pure preparation of peripheral blood mononuclear cells obtained by centrifugation of heparinized whole blood with a Ficoll/Triosil gradient. In 12 pairs studied within 5 yr of diagnosis, we observed a reduction in the percentage of cells reacting with OKT8 (recognizing the CD8 antigen present on the suppressor/cytotoxic subset) (P less than .05), but a similar level was detected in their nondiabetic cotwins. In 12 pairs studied greater than 5 yr after the diagnosis and in whom the nondiabetic twin is less likely to develop diabetes, the percentage of cells reacting with OKT8 was reduced in both the diabetic (P less than .05) and the nondiabetic (P less than .01) twins. Reductions were also seen with OKT3 (recognizing the CD3 antigen present on the total T-lymphocyte population) and OKT4 (recognizing the CD4 antigen present on the helper/inducer subset), but only in the diabetic twins from the group with longer discordance. We conclude that a reduced percentage of suppressor/cytotoxic cells is associated with type I diabetes, but the reduction appears to be genetically determined. Total T-lymphocytes are also reduced but mainly in the helper/inducer subset and only in diabetic patients of long duration. Such a reduction cannot therefore be primarily genetically determined. PMID- 2972576 TI - Clarification of signaling pathways mediated by insulin and insulin-like growth factor I receptors in fibroblasts from patients with specific defect in insulin receptor. AB - Receptor binding and biological action of insulin and insulin-like growth factor I (IGF-I) were studied in fibroblasts from a patient with leprechaunism and a patient with type A syndrome of insulin resistance. Insulin binding was reduced to 18.8 and 27.7% of control value, respectively. In contrast, IGF-I binding was normal in both patients. In competitive binding studies, IGF-I had 0.2% of the ability of insulin to compete with 125I-labeled insulin binding, and insulin had 0.1% of the ability of IGF-I to compete with 125I-labeled IGF-I binding in control subjects and patient fibroblasts. The dose-response curves of insulin stimulation assessed by glucose incorporation and alpha-aminoisobutyric acid uptake showed normal responsiveness, and ED50 was significantly shifted to the right in fibroblasts from both patients. However, normal responsiveness and sensitivity were observed in thymidine incorporation studies. For IGF-I, dose response curves of glucose incorporation, alpha-aminoisobutyric acid uptake, and thymidine incorporation were all normal in both patients. These results indicate that 1) the defect is specific to the insulin-receptor binding in these patients, 2) insulin and IGF-I activate glucose incorporation and alpha-aminoisobutyric acid uptake mainly through their own specific receptors, but 3) the IGF-I receptor appears to have a more important role in stimulating thymidine incorporation than the insulin receptor in physiological condition or, alternatively, an unknown postreceptor process with cascade signal transmission may overcome the decreased insulin-receptor binding to produce a normal dose response curve. PMID- 2972577 TI - Control of glucose metabolism in pancreatic beta-cells by glucokinase, hexokinase, and phosphofructokinase. Model study with cell lines derived from beta-cells. AB - Glucose usage by soluble fractions of cell extracts from two insulin-producing cell lines, RINm5F and HIT, was investigated. Analysis of enzyme activities indicated that glucose phosphorylation and phosphofructokinase are likely to be the rate-limiting steps of glycolysis in both RINm5F and HIT cell extracts. RINm5F extracts, which lack glucokinase, exhibited relatively flat concentration dependency curves of glucose usage and showed substantial inhibition of hexokinase. HIT cell extracts, which contain glucokinase but lack hexokinase, exhibited sigmoidal concentration-dependency curves of glucose usage, reflecting almost fully expressed glucokinase activity. A reconstituted system prepared from RINm5F and HIT cell extracts exhibited a composite concentration-dependency curve of glucose usage and showed substantial inhibition of hexokinase and almost fully expressed glucokinase. However, conditions that activate phosphofructokinase, such as addition of ammonium sulfate or fructose 2,6-bisphosphate or alkalization, removed the inhibition of hexokinase without noticeably affecting the glucokinase component of usage. Results obtained with a reconstituted system containing RINm5F cell extract and purified glucokinase were consistent with these findings. The data presented here indicate that this reconstituted cell free system serves as a valid model for the study of aspects of glycolytic control in the islet. This model illustrates the preeminent role of glucokinase in the control of glycolysis, consistent with its glucose-sensor function in the islet. In addition, these studies help to define the contribution of phosphofructokinase to the control of glycolysis and the mechanism whereby changes in phosphofructokinase activity could modulate, via changes in the glucose 6-phosphate concentration, the activity of hexokinase and hence the net glycolytic flux. PMID- 2972578 TI - Protein kinases 1988: a current perspective. AB - This review focuses on several recent developments in the field of protein kinases. In the area of protein serine/threonine kinases, much has been learned recently about protein kinase C structure and function. Novel lipid mediators, both stimulatory and inhibitory, have been discovered, and kinase has been shown to be an increasingly large family of gene products. Heterogeneity of cellular localization and function has been documented. Calcium/calmodulin-dependent protein kinases are now believed to consist of at least five enzymes, which range from those with extreme substrate specificity such as phosphorylase kinase and myosin light-chain kinases to calcium calmodulin kinase II, with several known substrates. Several of these enzymes appear to be important in synaptic transmission and, for calcium/calmodulin kinase III, in the regulation of protein synthesis. Several new examples of pseudosubstrate prototopes as endogenous kinase inhibitors have been described, including regions intrinsic to kinase primary sequences, which could serve as constitutive inhibitors of enzyme activity. In the field of protein tyrosine kinases, new enzyme species are being discovered at a rapid rate. There are several well-documented examples of kinase autophosphorylation on tyrosine leading to stimulation of catalytic activity. For the growth factor receptors with intrinsic protein tyrosine kinase activity, it now seems clear that kinase catalytic activity is necessary for most hormone effects on cells, with the general exceptions of ligand binding and, possibly, receptor cycling. Finally, several groups have recently described a close association between protein tyrosine kinases and a phosphatidylinositol kinase activity, a link that might eventually explain some of the initial steps in signal transduction that occur after kinase activation. PMID- 2972580 TI - [Left ventricular hypertrophy in hypertension: functional aspects]. AB - The purpose of this study was to detect the anatomical and functional response of the left ventricle in essential, uncomplicated hypertension. Fifty outpatients, whose hypertension (DBP greater than 95 mmHg on three separate visits) had been documented from few weeks to a maximum of 5 years, showing neither electrocardiographic abnormalities nor other signs referable to target organ damage, underwent an M-mode, computerised echocardiographic study. Twenty-seven patients has never been treated, and 23 patients were withdrawn from previous treatments for at least one month. Echocardiographic data of the left ventricle were compared with those of 38 normals, matched by sex, age and body surface area. Hypertensives showed a significant increase in both LV mass index (p less than 0.001) and relative wall thickness (p less than 0.001), a pattern of concentric hypertrophy, which prevented an inappropriate rise in wall tension. Twenty-four patients, whose LV mass was above the mean normal value plus two standard deviations, were considered to have left ventricular hypertrophy. The remaining 26 patients were classified as non hypertrophic, for their mass resulted lower than the mean value of controls plus 2 SD: left ventricular mass was 152 +/- 32 g/m2 and 97 +/- 19 g/m2, respectively (p less than 0.001). Systolic performance was enhanced in both subgroups of hypertensives, since their values of fractional shortening and those of mean-Vcf were both above the line which represents the normal relationship with end-systolic stress. Diastolic relaxation, evaluated by means of time to peak filling rate, was impaired only in the hypertensive subjects with increased LV mass (p less than 0.001 vs. normotensive), being retained in the non-hypertrophic patients. In the whole hypertensive group, LV mass index poorly correlated with casual systolic blood pressure (r = 0.31; p less than 0.05), while time to peak filling rate showed a good correlation with LV mass index (r = 0.76; p less than 0.001). We conclude that: 1) in essential hypertension without electrocardiographic abnormalities or target organ damage, echocardiographic left ventricular hypertrophy is a rather frequent finding; 2) the pattern of concentric hypertrophy, by reducing the increase in wall tension due to pressure load, contributes to the retained, or enhanced, systolic performance seen in our series; 3) on the other hand, left ventricular filling seems to be impaired in relation to the degree of myocardial hypertrophy. PMID- 2972579 TI - Regulation of the systemic immune response by visible light and the eye. AB - The injection of certain antigens into the anterior chamber (AC) of the eye results in the induction of antigen-specific suppressor T cells (Ts cells), which inhibit systemic delayed-type hypersensitivity (DTH). We have previously shown that down-regulation by Ts cells after AC injection with 2,4,6-trinitrophenol (TNP)-coupled spleen cells (TNP-Spl) is initiated by the intraocular activation of Ts inducer cells. These cells activate T suppressor-effector cells in the spleen that are responsible for suppressed DTH. With dark- and light-reared mice (Balb/c), we show that visible light has a direct effect on the intraocular T cell reaction that leads to systemic suppression. Our results show that if light is prevented from reaching the eye by dark rearing, by placing light-reared animals in the dark after AC injection, or by closing the eyelids of light-reared animals after AC injection, Ts cells are not activated. We show that light is responsible for establishing conditions in the eye that cause the preferential activation of Ts cells. The intraocular conditions established by light are not developmentally mandated as is visual development, but can be eliminated in adult light-reared animals by placing them in the dark for 18 h after AC injection. These conditions can also be induced in adult dark-reared animals by returning them to the light for just over 24 h before AC injection. These studies have important implications for understanding intraocular immune responses and possibly for the treatment of eye disease. PMID- 2972581 TI - [Supravalvular aortic stenosis. Analysis of a surgical series of 8 patients]. AB - Supravalvular aortic stenosis (SAS) is an unusual form of progressive left ventricular outflow tract (LVOT) obstruction, frequently associated with others vascular and physical anomalies. Between July 1980 and July 1986 eight such patients have been operated upon. In seven patients patch aortoplasty (according to McGoon or Doty technique) relieved the stenosis; in one, a two years old boy, with associated severe hypoplasia of the ascending aorta and aortic arch, a left ventricle (LV) - abdominal aorta valved conduit was associated to aortoplasty. One patient died in the operating room with a severe anterior myocardial infarction; seven patients are in good clinical condition at a mean follow-up of 32 months. The aortic supravalvular gradient was dramatically reduced in all patients according with Doppler echocardiography. In our mind, patients with SAS would benefit from an early surgical repair, because of progressive involvement of the aortic valve, coronary arteries and LV. The surgical technique is strictly related to the anatomical situations. Per and postoperative management should rely on the pathophysiology peculiar to this malformation (especially about coronary perfusion). Taking care to these points, the surgical results at a mid term follow-up are now very satisfactory in this cardiac malformation. PMID- 2972582 TI - New developments in pediatric interventional heart catheterization. PMID- 2972583 TI - Plasma human atrial natriuretic factor in cirrhosis and ascites with and without functional renal failure. AB - Functional renal failure of cirrhosis (FRFC) is a usually fatal syndrome of acute renal failure occurring in patients with advanced liver disease. Although not conclusively proven, most evidence suggests that renal arterial and arteriolar vasoconstriction is the cause of the renal failure in these patients. However, the mediators of the vasoconstriction remain unknown. Human atrial natriuretic factor (hANF) is a hormone with potent natriuretic, diuretic, and vasorelaxant properties. A deficiency of hANF could lead to renal arterial vasoconstriction and avid renal sodium retention as seen in FRFC. This study was undertaken to determine if patients with FRFC are deficient in circulating hANF. Seven patients with advanced alcoholic liver disease and renal failure of unknown cause (FRFC) were compared with 7 patients with advanced alcoholic liver disease, ascites, and normal serum creatinine as well as with 14 healthy volunteers. Plasma hANF was measured by radioimmunoassay. Plasma hANF was 742 +/- 227 pg/ml (mean +/- SEM) in patients with FRFC compared with 360 +/- 70 pg/ml in patients with liver disease and normal serum creatinine (p greater than 0.05) and 28 +/- 5.7 pg/ml in healthy volunteers (p less than 0.005 vs. FRFC and chronic liver disease, ascites, and normal serum creatinine). Thus, FRFC is not caused by a deficiency of circulating hANF. The elevated plasma hANF levels in patients with chronic liver disease and continued sodium retention may suggest a renal insensitivity to the natriuretic effects of hANF. PMID- 2972584 TI - Pathogenesis of sodium retention complicating cirrhosis: is there room for diminished "effective" arterial blood volume? AB - We measured total blood volume (125I-albumin), cardiac dimensions and function (echocardiography with Doppler), systemic hemodynamics (blood pressure and pulse), and plasma renin activity and norepinephrine levels in cirrhotic patients with and without ascites to assess the likelihood that either diminished central or arterial filling is the stimulus to sodium retention. Patients with ascites (n = 9) had significantly increased total blood volume, cardiac output, pulse rate, plasma renin activity, and plasma norepinephrine concentration, as well as decreased systemic vascular resistance and arterial blood pressure compared with patients without ascites (n = 8). Left atrial size was similar in the two groups but significantly larger than in normal control subjects. Right and left atrial pressures were also similar in retrospectively studied patients with and without ascites. Sodium retention in cirrhosis is probably not triggered by diminished central filling. Increased blood volume is compatible with either a primary hepatorenal stimulus to sodium retention or a signal arising from a region of underfilling within an otherwise expanded circulation. If the latter model is correct, a "hyperdynamic" systemic circulation and increased plasma neurohormone concentrations may indicate "effective" arterial underfilling in patients with ascites. PMID- 2972586 TI - [Open laparoscopy in patients at risk]. AB - A variant of the technique of open laparoscopy is reported. The principal feature of this technique is that creation of a pneumoperitoneum is simplified. No additional instruments are required, nor sharp instruments, except for the skin incision and splitting of the fascia. The modified open laparoscopy technique was used on 23 high-risk patients in a group of 224 patients on whom laparoscopy was performed at Ludwigshafen Gynecological Clinic. The technique is discussed with reference to the literature. PMID- 2972585 TI - Tuberculous peritonitis: laparoscopic diagnosis of an uncommon disease in the United States. PMID- 2972587 TI - [Donor erythrocyte typing as a possible method for preventing post-transfusion complications]. PMID- 2972588 TI - [Demographic genetic characteristics of the population of a large multiethnic city (the example of Alma-Ata). An analysis of assortative mating and migrations]. AB - A demographic genetic investigation of migration and marriage structure of the population of Alma-Ata was carried out on the basis of total analysis of marriage certificates issued in 1954 and 1984. The marriages were analysed in relation to birth places and nationality of married couples. An increase in the quantity of international marriages was noted, from 20.77% in 1954 to 22.85% in 1984 (p less than 0.05) in the population studied. The rate of international marriages for various nationalities was determined. Positive assortative mating, according to national indications, was proved in all basic national groups of the population. Negative correlation between the size of definite national groups and the coefficient of assortative mating was found. The quantitative and territorial geographical characteristics of migration are presented. The proportion of married natives of Alma-Ata was 10.7% in 1954 and 34.4% in 1984, the increase being significant. The territorial-geographical character of migration also changed: so, in 1954 the basic flow of migrants came from Siberia and the European part of the USSR, while in 1984 they came from neighbouring regions of Alma-Ata. The migration activity both of men and women was identical. Thus, the shortening of migration radius and lowering of migration intensity are marked. The population of the large multinational cities is proved to be not panmictic. PMID- 2972589 TI - [Medico-genetic study of the population of Kostroma Province. VI. The parameters of isolation by distance in the populations of the Bui and Shar'ya Districts of Kostroma Province]. AB - A model of isolation by distance proposed by Malecot and developed by Morton is applied to the data on marriage distances collected in two regions of Kostroma Province. There is good agreement between the estimates of local inbreeding when using the isonymy method and the model of isolation by distance. Interpopulation kinship approaches 0 at the distance 700 km. The mean coefficient of kinship for parents in the families with autosomal-recessive pathology is 20 times higher than mean coefficient of kinship in the population. PMID- 2972590 TI - [Effect of iodine vapors on the oral mucosa of workers in iodine manufacture]. PMID- 2972591 TI - [Sanitary improvement in the working conditions at the core departments of foundries in relation to the use of synthetic resins]. PMID- 2972592 TI - [Central hemodynamic function of miners operating underground self-propelled equipment in the Arctic Kola Peninsula]. PMID- 2972593 TI - Fatty streaks and fibrous plaques in human aorta show increased plasminogen activator activity. AB - Marked changes of the plasminogen activator activity, plasmin inhibition, and plasminogen activator inhibition were found in fatty streaks and fibrous plaques compared to the normal intima of the human aorta. Plasminogen activator activity and plasmin inhibition showed the following pattern: fibrous plaques greater than fatty streaks greater than normal intima. Plasminogen activator inhibition showed the opposite pattern: normal intima greater than fatty streaks greater than fibrous plaques. The tissue-type, but not the urokinase-type, plasminogen activator was detected immunologically. PMID- 2972594 TI - Effectiveness, specificity and safety of intranasal 1-deamino-8-D-arginine vasopressin treatment of normal blood donors. AB - A study of the absorption of 300 micrograms of 1-deamino-8-D-arginine vasopressin (DDAVP) given intranasally to normal blood donors was carried out to determine (a) the correlation between plasma levels of DDAVP and the percent rise of factor VIII procoagulant activity (VIII:C) and (b) the efficacy, specificity and safety of this treatment in increasing the recovery of factor VIII:C in donated blood. The maximum drug concentration was highly correlated to the maximum percent rise of VIII:C (r = 0.858, p less than 0.01). A differentiated effect of DDAVP on increases of VIII:C, VIII:Ag, vWF:Ag and vWF multimers was observed. A transient rise of fibrinopeptide A from 5 to 16 ng/ml, 30 min post-DDAVP, was not accompanied by changes in fibrinogen levels or generation of detectable factor Xa or thrombin. DDAVP had no effect on the factor XII-dependent pathway of plasminogen activation, or on the donor's vital signs and hematological parameters. Side effects were minor and of short duration. Intranasal DDAVP treatment of blood donors is considered to be a practical means of improving the recovery of VIII:C from normal donors. PMID- 2972595 TI - Two-year carcinogenicity study on three aromatic epoxy resins applied cutaneously to CF1 mice. AB - The cutaneous and systemic carcinogenic potentials of pure and two technical diglycidyl ethers of bisphenol A, DGEBPA, EPON 828 and EPIKOTE 828, respectively, have been investigated in six groups each of 50 male and 50 female CF1 mice. Twice weekly over a period of 2 yr, 0.2 ml of a 1 or 10% (w/v) solution of one of the epoxy resins in acetone was applied to the dorsal skin. A group of 50 male and 50 female CF1 mice was similarly treated treated with 2% (w/v) beta propiolactone in acetone (the positive control) while a group of 100 male and 100 female mice was treated with acetone alone (negative control). Survival of the CF1 mice to 2 yr was unaffected by cutaneous exposure to each epoxy resin. The compounds proved to be mildly irritant to murine skin, the response in males being greater than in females. There was a very low incidence of benign and malignant tumours of the skin and subcutis after exposure to any of these compounds. The number of systemic lymphoreticular/haematopoietic tumours was increased only in females treated with EPIKOTE 828 or DGEBPA. In male mice, treated with 10% EPON 828, there was a slight increase in the number of renal tumours. The incidence of other systemic tumours in either sex was not increased following cutaneous application of the purified or the two technical DGEBPA resins (1 to 10% in acetone). The significance of all these findings is fully discussed. PMID- 2972596 TI - [Bronchial obstruction--therapy with ipratropium bromide. Results of a multicenter clinical study]. PMID- 2972597 TI - [Enoximone, a new phosphodiesterase inhibitor: the spectrum of applications during heart surgery--a comparison with dobutamine]. AB - During cardiac surgery treatment of deterioration of myocardial function is usually based on catecholamines. Development of selective phosphodiesterase-(PDE )III-inhibitors seems to be a new aspect in treating myocardial dysfunction. Therefore the hemodynamic effects of the new PDE-inhibitor enoximone were investigated in 20 coronary surgery patients unable to be weaned from extracorporeal circulation (ECC) without pharmacological intervention (MAP less than 60 mmHg, CI less than 2.00 l/min.m2, PCP greater than 15 mmHg). After controlled reperfusion with 2.4 1/min.m2 two groups were separated in a random sequence receiving either 0.5 mg/kg enoximone as a bolus (n = 10), or dobutamine (n = 10, 5 micrograms/kg.min) as perfusion. In the dobutamine-group MAP and CI ( 14%) were decreased, while HR was increased significantly (+30%). Application of enoximone was followed by a slight increase in CI (+5%), a significant decrease in TSR while HR remained almost unchanged. PCP, too, differed significantly between the groups (enoximone: -38%; dobutamine: -10%). Ten minutes after weaning from ECC additional pharmacologic therapy (calcium, vasodilators, epinephrine) was necessary in eight dobutamine treated patients in contrast to four patients in the enoximone group (calcium, epinephrine). In patients with impaired myocardial performance during weaning from ECC enoximone seems to be an alternative therapy and is judged to be of some advantage compared to dobutamine application in this situation. The mechanism for improvement appears to be enhanced contractility owing to its positive inotropic effects, as well as a decrease in left ventricular outflow resistance resulting from peripheral vasodilation. PMID- 2972598 TI - Interferon-gamma receptors on T cells in patients with chronic liver disease. AB - Interferon-gamma (IFN-gamma) appears to be important for the activation of T cells, and its binding to IFN-gamma receptors on T cells is an essential step for its actions. We investigated the expression of IFN-gamma receptors on T cells in chronic liver disease using radioiodinated recombinant interferon-gamma binding assay, and by Scatchard analysis. The mean numbers of IFN-gamma receptors on T cells from controls, asymptomatic hepatitis B virus carriers (ASC), patients with chronic active hepatitis (CAH), and patients with liver cirrhosis (LC) were 2,205 +/- 497, 2,494 +/- 1,074, 1,925 +/- 735, and 1,666 +/- 653, respectively. The numbers of IFN-gamma receptors on T cells from the patients with LC were significantly smaller than those of controls (P less than 0.05). Kids of IFN gamma receptor on T cells from control and patient groups were 2.3 4.8 x 10(-9) M, and there was no significant difference among the groups. The percentage of T cells reactive with OKT3, OKT4, or OKT8 was similar in control and patient groups. These findings suggest that the decrease in IFN-gamma receptors in LC is not related to the activity of the liver damage, but is associated with the severity of the underlying disease. The normal expression of IFN-gamma receptors on T cells from CAH may provide a reasonable basis for IFN-gamma therapy to type B CAH. PMID- 2972599 TI - Angiofollicular lymph node hyperplasia associated with myelofibrosis case report and immunologic marker studies. AB - A case of angiofollicular lymph node hyperplasia (AFH) (hyaline vascular type) of the mediastinum associated with myelofibrosis is reported with immune marker studies. We hypothesize that the myelofibrosis is more than a mere coincidence since other myeloproliferative disorders have been observed in conjunction with AFH. Immunohistochemistry on cryostat sections of lymph node using the biotin avidin fluorescence technique showed the presence of numerous T helper cells and very few T suppressor/cytotoxic cells in the parafollicular areas. Concentric rings of lymphocytes showed the phenotype of mantle zone lymphocytes (i.e. IgM, IgD positivity), while the germinal centres were ill-formed as shown by Leu 14 (a Pan B cell marker) staining. By electron microscopy, follicular dendritic reticulum cells were readily found. The above findings suggest an abnormality of germinal center formation. PMID- 2972600 TI - Receptor-dependent transport of lysosomal enzymes. PMID- 2972602 TI - A novel mechanism for the selection of isotype-specific antibody responses: the role of intestinal T cells in the regulation of IgA synthesis by the anti suppressor circuit. AB - Within 6 hr of immunization the serum of mice contains a unique form of processed antigen, which consists of a complex of immunoglobulin (Ig) and antigen formed in the presence of a factor derived from the anti-suppressor inducer T cell. This complex binds to and activates the anti-suppressor effector T cell, which eventually leads to the inhibition of suppressor cell function. Both of these cells are present in the spleen (SPL) and play a role in the regulation of antibody responses. The purpose of these studies was to identify the anti suppressor T cells in gut-associated lymphoid tissue and compare their function to their splenic counterparts. Inducer cells were detected in the Peyer's patches (PP), mesenteric lymph nodes but not in the intra-epithelial lymphocytes. The effector cells, which take up the complexes, were detected in PP and lamina propria lymphocytes but not in the intraepithelial or mesenteric lymph node lymphocytes. Furthermore, the uptake of the complexes correlated with the presence of T cells bearing Ia antigens. The PP and SPL anti-suppressor cells were compared for their ability to enhance the production of IgA and IgG. The data clearly showed that the product of the inducer cell, and the effector cell it activates, not only enhanced the antigen-specific responses but also selected for isotype-specific antibody responses. Cells from SPL enhanced IgG greater than IgA, whereas cells from PP selected for IgA. Thus, the presence in PP of cells in the anti-suppressor circuit and their ability to selectively promote IgA synthesis suggest that this regulatory mechanism plays a significant role in intestinal immune responses. PMID- 2972601 TI - Human thymic dendritic cells. Characterization, isolation and functional assays. AB - The phenotypic analysis of human thymic dentritic cells (DC) in culture and in purified suspensions has been studied with light and electron microscopic (EM) immunolabelling techniques. Using a series of monoclonal antibodies (mAb) and a protein A-gold technique, we demonstrated that DR- and T6-positive cultured DC strongly bind the 9.3F10 mAb, an anti-DR-related antibody produced against human blood DC, and weakly express the T4 antigen, a membrane marker shared by Langerhan's cells (LC). On the other hand, thymic-cultured DC are negative for the other T-cell and monocyte-macrophage antigens. These results support the hypothesis that human thymic DC may be related to blood DC and epidermal LC. Moreover DC, unlike thymic macrophages, do not phagocytose latex particles, opsonized sheep red blood cells (SRBC) or Candida albicans. An efficient two-step technique of isolation, using a Percoll density gradient followed by an indirect panning technique, yields a purified (70-80%) thymic DC population, OKIa1-, 9.3F10- and OKT6-positive and esterase-negative. Immunolabelling and electron microscopy confirm that these isolated DC present similar phenotypic and ultrastructural features to human thymic DC in situ and in culture. Purified DC, used as stimulator cells in mixed leucocyte reaction (MLR), induce stronger proliferative responses than peripheral blood monocytes used as a control; blocking assays with OKIa1 mAb plus complement greatly reduced this stimulatory potency. These functional assays demonstrate that we obtained a purified typical DC population that can be used in immunological functional studies to elucidate the specific role of DC in human thymus. PMID- 2972603 TI - Interleukin-2 production and response by helper T-cell subsets in man. AB - This study investigated the production of, and response to, the lymphokine interleukin-2 (IL-2) by functional subsets of human CD4+ T lymphocytes. Fresh peripheral blood mononuclear cells (PBMC) were sorted into CD4+2H4+/4B4- suppressor-inducer cells, and CD4+2H4-/4B4+ helper cells. The suppressor-inducer subset proliferated well in response to the T-cell mitogens concanavalin A (Con) A and phytohaemagglutinin (PHA), and produced IL-2. The helper cells produced no detectable IL-2 and proliferated poorly. However, the latter population were induced to express functional IL-2 receptors by Con A or purified protein derivative (PPD), and proliferated well if supplied with exogenous rIL-2. These findings suggest that the two functional CD4 subsets are not independent, or counteracting, but rather that the generation of T-cell help is likely to involve cooperative interactions between the two subsets. PMID- 2972604 TI - Cyclophosphamide (Cy)-facilitated adoptive immunotherapy of a Cy-resistant tumour. Evidence that Cy permits the expression of adoptive T-cell mediated immunity by removing suppressor T cells rather than by reducing tumour burden. AB - A cyclophosphamide (Cy)-resistant immunogenic tumour, the L5178Y lymphoma, was used to demonstrate that Cy-treatment of a host bearing this tumour enables passively transferred tumour-sensitized T cells to cause complete tumour regression without any need for Cy to cause a reduction in tumour burden. It was shown that whereas infusion of tumour-sensitized T cells from immune donors had very little effect on growth of the tumour, and whereas treatment with 150 mg/kg of Cy caused appreciable enhancement of tumour growth, combination therapy with Cy plus immune T cells caused complete tumour regression and resulted in long term survival. Evidence that Cy treatment facilitated the expression of adoptive immunity against the L5178Y lymphoma by eliminating tumour-induced suppressor T cells consisted of the demonstration that tumour regression caused by combination treatment with Cy and immune T cells could be inhibited by infusing the recipient with Cy-sensitive, L3T4+ T cells from tumour-bearing but not from normal donors. PMID- 2972605 TI - Maternal autoimmune disease influences self-tolerance in offspring: the role of suppressor cells and materno-foetal cell traffic. AB - Autoimmune haemolytic anaemia (AIHA) was induced in normal strain (CBA/Ca/T6) mice by repeated intraperitoneal injection of rat red blood cells (RRBC). Antibody production to cross-reactive antigens on mouse red blood cells (MRBC) and foreign antigens on RRBC was measured by the direct antiglobulin test (DGAT) and serum haemagglutination, respectively. RRBC primed female or male mice and sheep red blood cell (SRBC) primed controls were mated with naive partners and their progeny immunized with RRBC in adult life. The offspring of mothers but not fathers with active autoimmune disease showed a significant reduction in antibody response to self (MRBC) antigens, whereas the response to non-self (RRBC) was unaffected. Transfer of 30 X 10(6) spleen cells from the progeny of RRBC primed mothers into non-irradiated normal recipients resulted in selective suppression of the anti-self response following challenge with RRBC, provided that the cell donors had been boosted with RRBC 7-10 days before the transfer was performed. Thus the progeny of mothers with AIHA possessed self-reactive memory suppressor cells (Ts) shown previously to belong to the Thy-1+ I-J+ Ly-2+ T cell subset in this model. To test whether the Ts were of maternal or foetal origin the suppressor assay was repeated with spleen cells from the F1 offspring of RRBC primed B10.A(3R) (I-Jb) mothers and normal CBA(I-Jk) fathers. Pretreatment with anti-I-Jb serum plus complement completely abrogated suppression on adoptive transfer but anti-I-Jk serum failed to do so, indicating that the Ts were derived from the mothers. These findings emphasize the potential importance of Ts in induction of self tolerance during early ontogeny. PMID- 2972606 TI - Inhibition of cell-mediated immunity by tumour cell products: depression of interleukin-2 production and responses to interleukin-2 by mouse spleen cells. AB - Supernatants from cultures of mouse and human tumour cells inhibited the production of interleukin-2 (IL-2) by stimulated mouse spleen cells. The tumour cells tested, all of which were active, included a mouse and a human melanoma, three methylcholanthrene-induced fibrosarcomas of mice, and human HeLa cells. Supernatants from normal mouse and human fibroblasts were inactive. Inhibition was dose-dependent. Spleen cells from aged mice were more susceptible to inhibition than spleen cells from young mice. When tumour cell culture supernatants were fractionated on Sephacryl S-300, two peaks of activity were found, with apparent molecular weights of approximately 50 and 18 kD. Supernatants from tumour cell and fibroblast cultures caused variable, but generally weak, inhibition of responses of lymphoblasts to IL-2. It is suggested that inhibition of IL-2 production may be an important mode of action of tumour cell products that inhibit cell-mediated immunity. PMID- 2972607 TI - [Psoriasis-like eruption caused by flecainide]. PMID- 2972608 TI - [Immunologic changes and treatment with cyclosporin A in a case of erythrodermic psoriasis]. PMID- 2972609 TI - Immunoregulation of lymphoproliferation in vitro by monocytes and their subpopulations. I. The induction of suppressor T cells in long term cultures and the role of MHC class II determinants and preliminary characteristics. AB - Peripheral blood monocyte (MO) subpopulations isolated on a basis of functional expression or a lack of Fc receptor (FcR+ and FcR- MO) were used to study the regulation of the antigen (PPD)-driven lymphoproliferation. Long-term cultures of T lymphocytes with FcR+ MO, but not FcR- MO, led to the induction of T suppressor (Ts) cells that inhibited antigen-driven lymphoproliferation in the test cultures. These Ts cells were resistant to mitomycin C, belonged to afferent acting category of Ts cells, expressed CD8 and HLA-DR determinants, and showed no antigenic specificity nor genetic restriction in action. The expression of MHC class II molecules (HLA-DR and HLA-DP but not HLA-DQ determinants) on MO which were used for antigen presentation was critical for Ts cell induction. It was concluded that specialized MO subpopulations may regulate the lymphoproliferation by inducing Ts cells (or Ts cell circuit) that in turn inhibit antigen-driven immune response. PMID- 2972610 TI - Effect of an autoreactive T cell clone from (NZB x NZW)F1 mice on the production of anti-DNA antibodies in vivo and in vitro. AB - An autoreactive T lymphocyte clone, designated as F1C4 was established from an autoimmune mouse strain (NZB/NZW)F1. This clone proliferated in the presence of mitomycin C-treated splenic adherent cells (MMC-SAC) from syngeneic mice. This response was dependent on the numbers of MMC-SAC. The specificity of F1C4 for I-A was determined by an inhibition test carried out with monoclonal anti-Ia sera. Furthermore, the F1C4 cells did not exhibit alloreactivity in a proliferation assay and did not react to foreign antigens such as fetal calf serum (FCA) used in the culture medium. When F1C4 cells were cultured with autologous non-T cells in the absence of antigen, they strongly enhanced IgM class anti-ssDNA production from non-T cells of both young and old B/W F1 mice at appropriate cell numbers in vitro. Furthermore, the production of IgG class anti-ssDNA from non-T cells of old B/W F1 mice was also enhanced. The adoptive transfer of F1C4 cells enhanced the levels of both IgM and IgG anti-ssDNA antibodies in the serum of aged B/W F1 mice. Moreover, the serum levels of anti-ssDNA of IgG2a and IgG2b subclasses were readily enhanced by the transfer of F1C4 in vivo. PMID- 2972611 TI - Cyclosporin A blocks the generation of alloindifferent but not of allospecific suppressive cells. AB - The induction of alloantigen-indifferent, MHC unrestricted suppressive cells (SC) early on in human mixed lymphocyte culture (MLC) or after stimulation with suppressive T cell clones was blocked in a dose-dependent fashion by cyclosporin A (CsA). This was not prevented by the addition of the following defined lymphokines: interleukin (IL) -1, -2, -4, -5, interferon-tau or GM-CSF. The addition of MLC-conditioned medium as a source of multiple lymphokines including IL-3 also failed to reconstitute suppressive activity in the presence of CsA. In contrast, the development of allospecific, HLA-restricted SC in the same CsA blocked MLC was not prevented or was even enhanced. These results confirm that CsA 'spares' specific SC induction late in MLC, but show that it prevents induction of non-specific suppression earlier in MLC by a mechanism presumably unrelated to blocking the secretion of interferon-tau or IL-1 to IL-5. PMID- 2972614 TI - Flow cytometric analysis of lymphocyte activation in the mixed lymphocyte response. AB - Interleukin 2 receptor (IL2R) expression may be a useful parameter for assessing lymphocyte activation in the mixed lymphocyte response (MLR). However, the contribution of irradiated stimulator (S*) cells to the levels of IL2R+ cells recovered must first be defined. We have used a flow cytometric parameter termed the sip count to assess this potential contribution of S* cells. This parameter, which is the number of cells within a defined cell gate, is a reflection of the viable cell number per culture well, since (a) a constant number of cells were plated per well on day 0, (b) cells recovered from a well were resuspended in a constant volume, (c) the flow cytometer aspirated (sipped) a constant volume of cell suspension, and (d) nonviable cells were not included in the gate. Sip count assessment showed that only 5% of S* cells were recoverable by day 4 of culture; in contrast, 70% or more of unstimulated responder cells were recoverable. Sip counts of MLR cultures identified an increase in cell number beginning on day 5, reflecting DNA synthesis and cell division. We then used the sip count to assess changes in the levels of IL2R+ cells in MLR cultures. The number of IL2R+ cells continued to increase up to day 7, even though maximal DNA synthesis occurred on day 5. Further, dual color analysis revealed that the proportion of CD4 cells expressing IL2R was maximal on day 5, whereas the proportion of CD8 cells expressing IL2R continued to increase until day 9. These findings show that flow cytometry can be used to study lymphocyte activation by alloantigens. PMID- 2972613 TI - A rat antibody against a structure functionally related to the mouse T-cell receptor/T3 complex. PMID- 2972612 TI - The Ly-10 antigen is a marker of mouse-activated T lymphocytes. AB - Ly-10.1 is a lymphocyte surface antigen controlled by a gene linked to the Ly-1.1 locus and expressed on activated T helper, T suppressor (Ts), and cytotoxic T lymphocytes (CTL). In this report, we describe the following: 1) Ly-10 is a heterodimeric glycoprotein consisting of a 80,000 heavy and a 34,000 light chain. 2) Although mature CTL are Ly-10+ by negative selection with anti-Ly-10.1 and complement (C), CTL precursors reactive to allogeneic cells are Ly-10-. 3) Similarly, IL-2-producing effector T cells induced by Mls-incompatible cells and semiallogeneic stimulation are eliminated by anti-Ly-10.1 and C after activation but are not eliminated as precursors before activation. 4) In mixed lymphocyte culture with semiallogeneic cells, the frequency of Ly-10.1+ cells was highest on the 2nd to 5th day after stimulation, decreased by the 12th day, and increased after restimulation with fresh antigen as demonstrated by immunofluorescence, C mediated lysis, and IL-2 production. 5) When spleen cells were treated with anti Ly-10 and C before concanavalin A (Con A) activation, the suppressive activity in the Con A T blasts was reduced, suggesting that in normal mice, some Ts preexist in a Ly-10+ activated state. These results indicate that Ly-10 is a marker of activation of T cells, not expressed on precursor T cells and whose expression is both transient and dependent on the presence of antigen. The similarities in biochemical and cellular characteristics suggest that Ly-10 is a mouse homologue of the human lymphocyte activation marker 4F2. PMID- 2972615 TI - Phage types of Staphylococcus aureus isolates from the throats of school children. PMID- 2972616 TI - Why the quick fix won't fix today's nurse shortage. PMID- 2972617 TI - The dynamics of health insurance loss: a tale of two cohorts. AB - Past research has characterized the uninsured population in fairly static terms. Little is known about the transition between insured and uninsured states, the length of time persons remain uninsured, the existence of multiple spells of health insurance loss, and the extent to which uninsured persons subsequently acquire private or public coverage. In this paper we examine these dynamic aspects of health insurance coverage for cohorts of privately insured and uninsured persons over a 32-month period. The uninsured population was found to be quite heterogeneous, consisting of many persons who lost coverage for relatively short periods of time, others who were periodically without insurance, and many who were persistently uninsured. Implications for public policy initiatives for the uninsured population are also discussed. PMID- 2972618 TI - The erosion of purchased health insurance. AB - In this paper, we trace the decline of purchased health insurance and examine the reasons for the rapid growth of self-insurance between 1981 and 1985. Then, using nationally representative data on benefits in larger private sector firms, we examine the changing content of self-insured plans and compare them with fully insured conventional plans from commercial insurers and Blue Cross and Blue Shield Plans. Between 1981 and 1985, the percentage of employees in mid- to large sized firms covered by self-insurance grew from 21% to 42%. Self-insured plans cost more than purchased plans in 1981, and continued to cost more in 1985. Their higher premiums were not due to richer benefit packages. Indeed, they less often covered "fringe" services and required greater cost sharing via higher deductibles and coinsurance. Upon considering both the efficiency and the equity issues of self-insurance, we sound a cautionary note on this growing trend. PMID- 2972619 TI - Uncompensated care pools and care to the uninsured: lessons from the New York Prospective Hospital Reimbursement Methodology. AB - One policy response to both the growing number of uninsured and the increased volume of uncompensated care is the development of revenue pools to explicitly finance uncompensated care. Despite the growing popularity of this response, few analyses have examined their success in financing uncompensated care and improving access to the uninsured. This study examines one such program developed as part of New York State's all-payer rate-setting system. The results indicate that the revenue pools improved the financial condition of New York hospitals but were less effective in improving access to care by the uninsured. The latter result was traced to the method selected by New York to measure "need" and distribute payments to hospitals. If the goal of such programs is to earmark payments to the uninsured, methods other than the New York system should be employed. PMID- 2972621 TI - Factors associated with Medicare beneficiaries' interest in HMOs. AB - In this study we explore the demographic and health status factors associated with HMO awareness and enrollment interest among 314 Medicare beneficiaries. The results reveal that older adults most aware of HMOs were more demographically advantaged than others. In contrast, those most interested in information and HMO enrollment reported more financial vulnerability. An unanticipated finding was that self-reported health status and health care utilization were not associated significantly with HMO awareness or enrollment interest. If replicated, the findings suggest that relatively healthy Medicare beneficiaries with financial vulnerabilities could be enrolled in HMOs without necessarily increasing the proportion of high medical care utilizers. PMID- 2972620 TI - The early effects of Medicare's prospective payment system on psychiatry. AB - In this paper, we study the effects on psychiatry of Medicare's prospective payment system (PPS) during 1984, as PPS was implemented. We examined data on psychiatric discharges before and after PPS from three kinds of hospitals--those with psychiatric units exempt from PPS, those with nonexempt units, and those that treated psychiatric patients in scatter beds--as well as data on all hospital discharges. We conclude that the providers of psychiatric services responded to the incentives inherent in PPS much the way hospitals as a whole did -with significantly reduced lengths of stay. Of the three kinds of hospitals that rendered psychiatric care, those that treated patients in scatter beds had the greatest reduction in length of stay. Using readmission rates as a gross indicator of quality, we conclude that quality did not suffer because of the shortened stays. PMID- 2972622 TI - Trends and regional variations in hospital utilization and quality during the first two years of the prospective payment system. AB - The Commission on Professional and Hospital Activities monitored both Medicare and non-Medicare hospital discharges for a cohort of 646 U.S. nonfederal, short term general hospitals from nonwaivered states over the period 1980-1985. Using this data base and a linear forecasting model, we studied utilization and quality of-care trends for the years preceding implementation of the prospective payment system (PPS) and the two years following its implementation. We found that Medicare discharges declined significantly in both 1984 and 1985, length of stay was unchanged from 1984 to 1985 (except in the West, where further declines occurred), and skilled nursing facility and home health care use for postdischarge care increased both years. Our unadjusted measures of quality showed no adverse effects in either 1984 or 1985 for Medicare patients. PMID- 2972623 TI - Cost and quality effects of outpatient cataract removal. AB - In March 1985, a Health Care Financing Administration regulation went into effect requiring that cataract removal without exceptional circumstances be done in outpatient settings. In this paper, we study implications of that mandate by comparing the cost and quality outcomes of cataract removal in outpatient and inpatient settings both before and after the regulation went into effect. After controlling for population and physician differences for both study periods, we found by chi square significantly fewer infections, suture adjustments, and pain requiring medication among outpatients than inpatients. Log-linear regression, however, found that the only significant predictor was inpatient or outpatient, and only for infection (p = .02), with an odds ratio of 7.55 (95% confidence interval; .92-61.60). We also found lower Medicare payments for outpatients in both study periods. For the preregulation study groups, inpatient care was 34.8% more costly than outpatient care; the cost differential dropped to 32.5% for the postregulation study groups. PMID- 2972624 TI - Hospital-physician integration and hospital costs. AB - The integration of physicians into the management and policy-making structure of the hospital has been widely touted as necessary to achieve organizational efficiencies in a changing health care environment. This paper examines the implications of five such integrative strategies for hospital costs. Our findings indicate that greater administrative participation by physicians is consistently related to higher costs. We also found that other integrative strategies are associated with higher hospital costs in specific types of hospitals. We discuss the implications of our findings for hospital managers and policy makers. PMID- 2972625 TI - Determinants of provider selection or assignment in a mandatory case management program and their implications for utilization. AB - Mandatory managed-care programs for Medicaid beneficiaries typically require enrollees to select the provider who will act as gatekeeper to the medical care system. A substantial number of beneficiaries, however, do not exercise this choice and are assigned a gatekeeper. Using consumer survey data from the Missouri Managed Health Care Project, we examined characteristics and use experiences of assignees compared with selectors. We found that the assignees enjoyed better health than the selectors and were less likely to have had a regular source of care prior to the program. The utilization experience was similar for both groups. We conclude that the basis for not making a choice is one of indifference. PMID- 2972626 TI - Ability of monophosphoryl lipid A to augment the antibody response of young mice. AB - Treatment with nontoxic monophosphoryl lipid A increased the magnitude of the immunoglobulin M (IgM) antibody response to type III pneumococcal polysaccharide in young (2- to 4-week-old) mice. This was accompanied by the appearance of significant numbers of IgG1- and IgG3- secreting antibody-forming cells in 4-week old mice. These findings indicate that monophosphoryl lipid A can be used as an adjuvant to improve the immunogenicity of poorly immunogenic antigens in young, immunologically immature animals. PMID- 2972627 TI - Virulence properties and enterotoxin production of Aeromonas strains isolated from fish. AB - The biological activities in vivo and in vitro of 59 motile Aeromonas spp. isolated from fish and water tanks were simultaneously analyzed in poikilothermic and homoiothermic systems. A total of 64.3% of the isolates tested were pathogenic for fish, and 62% of Aeromonas hydrophila and A. sobria isolates either virulent or nonvirulent for fish were enterotoxigenic. Although the majority of the strains were proteolytic and amylolytic and produced DNase, other activities, such as elastase and staphylolysis, were only present in A. hydrophila. Most of the strains (96%) produced hemolysins, and 68% had agglutinating capacity, but neither isolates pathogenic for fish nor enterotoxigenic isolates showed specificity for trout or human erythrocytes, respectively. The production of siderophores, agglutination in acriflavine, and precipitation after boiling were found not to be useful tests for screening virulent strains. Although statistical analysis revealed a significant relationship between virulence for fish and positive results for arabinose and sucrose fermentations, elastase, and hemolysis of human erythrocytes, only lysine decarboxylase showed a significant positive relationship with enterotoxigenicity. Using extracellular products from representative Aeromonas strains with different virulence markers and belonging to distinct O serogroups, we demonstrated a lack of correlation between cytotoxicity for fish and homoiothermic cell lines and pathogenicity. The extracellular products from selected pathogenic A. hydrophila strains were lethal for rainbow trout and displayed proteolytic, hemolytic, and cytotoxic activities which were simultaneously lost after heat treatment. The findings reported here indicate that it is not possible to establish a common and single mechanism involved in the invasion of Aeromonas spp. in poikilothermic and homoiothermic hosts. PMID- 2972628 TI - Hepatitis B vaccination with short dose intervals--a possible alternative for post-exposure prophylaxis? AB - To achieve a more rapid antibody response following hepatitis B (HB) vaccination, vaccine injections were given to medical students at considerably shorter intervals than usually recommended. They received 10 micrograms of the Merck Sharp & Dohme recombinant HB vaccine at time 0, 2 and 6 weeks (27 vaccinees) or were vaccinated according to the recommended schedule for pre-exposure HB prophylaxis (0, 1 and 6 months) (26 vaccinees). The short interval regimen resulted in a significantly higher frequency of protective antibody levels (greater than or equal to 10 IU/l) two weeks after the second dose of vaccine (48% vs. 4%; p less than 0.001), and all short interval vaccinees had seroconverted within two months (i. e. two weeks after the third dose). The recommended interval regimen resulted in a slower development of antibodies but significantly higher peak antibody levels after the completed three doses (p less than 0.001). The results indicate that protective antibody levels against hepatitis B virus (HBV) can be achieved more rapidly in humans through vaccination with short intervals. This short interval vaccination regimen, which has proved effective for post-exposure prophylaxis in chimpanzees, should possibly also be considered for post-exposure prophylaxis in humans, for instance after accidental exposure to HBV-contaminated blood. PMID- 2972629 TI - Proliferative and cytolytic potentials of purified human tumor-infiltrating T lymphocytes. Impaired response to mitogen-driven stimulation despite T-cell receptor expression. AB - Using limiting dilution analysis (LDA) we have previously shown that in most instances, the frequency (F) of proliferative T lymphocyte precursors (PTL-P) was strikingly reduced in tumor-infiltrating lymphocytes (TIL). In this study involving 19 cases, we show that the impaired clonogenic potential of CD2+ TILs is primarily caused by an intrinsic defect rather than to suppressor T cells or to a direct effect of the tumor cells usually present in the culture system. This was demonstrated by experiments in which the F of PTL-Ps was quantitated both in highly purified CD2+ TILs (using a cell-sorter) and in non-purified TIL suspensions (still containing tumor cells), which originated from the same biopsy specimen. The F of PTL-Ps was virtually identical in either sorted or nonsorted suspensions and the data from LDA were always consistent with the single-hit Poisson model, indicating that no suppressor cells interfered with growth of CD2+ TIL. Stimulation of sorted CD2+ TIL in low-density cultures by either phytohemagglutinin or anti-CD3-monoclonal antibody (MAb) indicated that the antigen-dependent activation pathway was impaired, although structurally intact T cell receptor (TCR) complexes were apparently expressed, as assessed by immunofluorescence. The depressed proliferative response of CD2+ TIL could not be reversed in vitro when phorbol-esters were used in combination with ionomycin, which bypass the TCR. Nevertheless, 180 clones obtained from 8 cases were analyzed for their cytolytic activity. The majority mediated specific lytic activity (against unknown antigens), as assessed by lectin-dependent cell cytotoxicity, whereas only 6% of them manifested lymphokine-activated killing on appropriate targets. PMID- 2972630 TI - Expression of the c-Ha-ras and neu oncogenes in DMBA-induced, anti-estrogen treated rat mammary tumors. AB - Dimethylbenzanthracene (DMBA)-induced rat mammary tumors were analyzed for the structure and expression of the oncogenes c-Ha-ras and neu and the effects of anti-estrogen treatment. Tumor samples were divided into 3 groups, the first consisting of untreated tumors, the second of anti-estrogen (toremifene)-treated unresponsive (growing) tumors, and the third of toremifene-treated responsive (regressing) tumors. DNA and RNA derived from normal tissues of the same experimental animals were also analyzed. In Southern blot analysis of genomic DNAs, 2 tumors out of 23 contained a new Xbal site in the Ha-ras gene, indicating a point mutation in the second nucleotide of codon 61. Both of these tumors belonged to the group that had not received toremifene. No amplifications of the Ha-ras or the neu genes were observed. Although greatly variable, the levels of Ha-ras mRNA were highest in untreated tumors, lower in toremifene-treated, unresponsive tumors and even lower in toremifene-treated, regressing tumors, corresponding approximately to the levels detected in normal liver and uterus of untreated animals. Expression of the neu mRNA was variable and considerably lower than that of Ha-ras mRNA. It was similar in all 3 groups and somewhat elevated than in several non-malignant control tissues. Localization of c-Ha-ras expression by in situ hybridization revealed a relatively even distribution of the mRNA throughout the mammary tissue. The results suggest that mechanisms other than activation of the c-Ha-ras or neu genes are important for progression and regression of DMBA-induced rat mammary carcinomas. PMID- 2972631 TI - Etofenamate and transcutaneous electrical nerve stimulation treatment of painful spinal syndromes. AB - Thirty patients suffering from painful syndromes of the spine were admitted to a randomized controlled clinical trial. They were divided into two groups and treated either with transcutaneous electrical nerve stimulation (TENS), one application every other day, for 20 days or with TENS and an ointment containing etofenamate 10% gel, 3-5 cm daily on the day of TENS therapy, and the same dose twice daily on the other days. The associated therapy achieved, when compared with TENS alone, a statistically significant better outcome. Furthermore a marked improvement of symptoms was observed in a shorter period of time. Therapy was well tolerated and in only four cases mild, self-limiting, skin reactions were observed. On the basis of these results the use of etofenamate and TENS could represent a viable alternative to systemic nonsteroidal antiinflammatory drug therapy. PMID- 2972632 TI - Prevention of low dose streptozotocin induced diabetes by muramyl dipeptide. AB - The objective of the present investigation was to evaluate the effect of the synthetic immunomodulator MDP on an experimentally induced diabetes. It has been previously demonstrated that a single high dose of streptozotocin (STZ) induces hyperglycemia by direct destruction of pancreatic beta-cells. MDP had no effect on the diabetes induced by high dose STZ injection. However, MDP partially protected mice against the toxicity of STZ. In contrast to the first model, repeated low dosages of STZ have been shown to induce hyperglycemia due to autoimmune destruction of beta-cells. Large dosages of MDP given before these low dosages of STZ markedly decreased the diabetogenic effect of STZ. It is proposed that this protection is due to the immunosuppressive activity of MDP. PMID- 2972633 TI - Effects of cyclosporin A on in vitro lymphocyte response to autologous or allogeneic stimulation: influence of HLA phenotypes. AB - In this study we investigated whether the interindividual variability of lymphocyte sensitivity to cyclosporin A (CsA) could be controlled by the HLA region. The models used were the in vitro primary and secondary autologous (AMLR) and allogeneic mixed lymphocyte (MLR) cultures of cells from 32 healthy subjects from our HLA reference panel. Our results show that CsA inhibited primary allogeneic MLR to a much greater extent than primary AMLR (-81 +/- 2% vs -38 +/- 8%, P less than 0.001). The same pattern was observed when cells harvested from CsA-treated primary cultures were rechallenged in secondary cultures with the original sensitizing stimulator cells (-40 +/- 6% vs -17 +/- 9%, P less than 0.05). No differences were observed in primary autologous and allogeneic cultures among responders of different HLA phenotypes. In contrast, the secondary responses did vary according to the HLA types: in secondary AMLR, CsA-priming did not lower, or even enhance, the proliferative responses of DR5+ and/or DR2+ lymphocytes (+7 +/- 13%), whereas it significantly lowered the responses of DR2-5 cells (-46 +/- 8%). In secondary MLR, lymphocytes proliferation was lowered by CsA-priming in all but DRW11(5)+ subjects (-45 +/- 7% vs +2 +/- 23%, P less than 0.05). It is concluded that the individual HLA phenotype influences the pattern of lymphocyte sensitivity to CsA. PMID- 2972636 TI - Some considerations on control of continuous processes by speech. PMID- 2972635 TI - Adapting the environment to people with disabilities: constitutional issues in Canada. AB - Many problems experienced by people with disabilities can be ascribed to the effects of a disabling environment rather than personal deficiencies. Adapting the environment to make it less handicapping therefore has become a significant item on agenda of reforms sought by people with disabilities, but pursuit of this objective has wider legal, political and constitutional implications. Canada is one country in which all of these avenues have been explored in order to achieve progress in this sphere. This article, based on interviews with key informants, reports the constitutional and historical background to the Canadian experience and identifies various problems that have been encountered by the organisation and persons who have sought to secure appropriate changes. It is argued that this particular example, founded on a hard-won clarification of rights and entitlements under the Constitution, may offer a more promising way forward than alternative approaches focused on legislative or administrative regulations. PMID- 2972634 TI - Androgenic-anabolic steroid effects on serum and skin surface lipids, on red cells, and on liver enzymes. AB - The effects of large doses of testosterone and anabolic steroids on the serum lipids and skin surface lipids were studied during a 12-week strength training period. Decreased serum high-density lipoprotein cholesterol (HDLC) (P less than 0.001) and elevated serum triglyceride (P less than 0.05) levels were found at the same time with an elevated skin surface lipid cholesterol level. The erythropoiesis and function of the liver were only slightly stimulated. These abnormalities were assumed to be a direct effect of the testosterone and anabolic steroids. PMID- 2972637 TI - Planning a community survey of the younger chronic sick and disabled living in Lothian region. PMID- 2972638 TI - Study of integration strategies. PMID- 2972639 TI - Behavior modification and therapy in rehabilitation. PMID- 2972640 TI - Behavioural assessments of physically disabled patients: agreement between rehabilitation therapists and nurses. PMID- 2972641 TI - Architects supporting the integration of the disabled within the community. PMID- 2972642 TI - Using handicap-specific data of disabled workers to select adequate working tasks. PMID- 2972643 TI - Methods of changing the social environment. PMID- 2972644 TI - Disability and the built environment. PMID- 2972646 TI - Psychological factors in back pain: self-rated back pain. PMID- 2972645 TI - The understanding and evaluation of chronic pain: subjective vs. objective determinants of pain. PMID- 2972647 TI - Social aspects of rehabilitation. PMID- 2972649 TI - The dynamics of vertebrogenic pain and autodermographic recording. PMID- 2972648 TI - Experience with minimal therapy of low back pain. AB - A group of 20 patients suffering from repeated low back pain with radicular symptomatology in area L5 or S1 was followed-up during an acute attack, without use of some kind of analgetics or only in very reduced quantity. The protective function of pain and especially low back pain had been explained to the patients. The patients were instructed to engage in all movements which do not provoke pain, or if so, at an easily bearable level. They may have to walk with a scoliotic posture or to crawl. The average time of vertebrogenic pain attacks treated with analgetics lasted longer than the ones with minimal drug therapy. PMID- 2972652 TI - Information technology and remote working for disabled people. PMID- 2972651 TI - Assistive devices--from prototype, to product, to users. PMID- 2972650 TI - A comparison of simulated and realistic emergency stop responses: relevance of driving assessment. PMID- 2972653 TI - Fitness for work: medicolegal assessment of residual disability and employment handicap. PMID- 2972654 TI - Systematic information within the European community's action programme on disabled people. PMID- 2972655 TI - Towards a national action plan to meet the changing needs of the disabled & handicapped persons. PMID- 2972656 TI - New trends in the adjustment of disabled people to the marketplace: a United States perspective. PMID- 2972657 TI - A medical view of research methodology in rehabilitation. PMID- 2972658 TI - A critical evaluation of car seat belts for disabled people: the problems of upper limb mobility and manual dexterity. PMID- 2972660 TI - Studies on the effects of the antiandrogen cyproterone acetate on social encounters between pairs of male mice. AB - An attempt was made in two experiments to reinvestigate the effects of the antiandrogen cyproterone acetate (CA) on mouse social behavior in a variety of ethologically-assessed paired encounters. The data confirm that CA reduces offense (threat and attack) in animals when both subjects receive the material but that CA has no such action in other pairings. This suggests that CA's major suppressive effect on "hostility" is expressed in mice via a reduction in "attack promoting" pheromone production. Indeed, there was evidence in the more chronic study that CA, could augment (via a central mechanism?) offense in subjects paired with docile anosmic opponents. Changes in defense were largely responses to variations in the degree of attack to which animals were subjected. The antihormone also had actions on other aspects of behavior including sexual activity, social investigation and immobility in such tests. CA had a potent suppressive action on the weights of sex accessory glands. The data do not suggest that CA can be used as a specific antihostility agent. PMID- 2972659 TI - Behavioral effects in adulthood of serotonin depletion by P-chlorophenylalanine given neonatally to male rats. AB - The effect in male rats of depletion of brain 5HT over days 1-7 or 8-16 after birth, by daily administration of 100 mg/kg PCPA was noted on various behaviors after the animals had reached adulthood. Treatment over days 1-7 had little or no effect, but over days 8-16 induced the following behavioral changes: In the elevated plus-maze, the treated group spent longer on the open arms, indicating a lower level of anxiety. In a test for sexual orientation, the treated animals were more sexually oriented toward the female stimulus and they showed a reduction in pendulum movements (a fear-related parameter); however their sexual performance with a receptive female was not different from the controls. The animals were also tested as residents in a resident-intruder paradigm for their social and agonistic activity. The treatment induced an increase in offensive activity confined to the high intensity elements and a parallel decrease in defensive activity. These results can be interpreted in terms of an increased reactivity to environmental and social cues and a decreased anxiety. PMID- 2972661 TI - The effect of ketoconazole on sebum secretion in patients suffering from acne and seborrhoea. AB - Ketoconazole, an imidazolic antimycotic derivative, has proved to have an antiandrogenous effect, which may be observed to inhibit the adrenal and gonadal steroid synthesis. The authors wished to verify this effect of the drug in men and women suffering from acne and seborrhoea, conditions believed to indicate a heightened androgenous effect in the sebaceous gland. They aimed to do this by studying the cutaneous sebum, which is considered a reliable indirect indicator of androgenous activity. They found a statistically significant decrease in the rate of sebum production in all patients administered with a single daily oral dose of ketoconazole (200 mg). Moreover no untoward side-effects were reported. The anti-androgenous property of ketoconazole would therefore suggest that the drug could be proposed as a useful alternative in the treatment of acne and seborrhoea. PMID- 2972662 TI - Topical spironolactone inhibits dihydrotestosterone receptors in human sebaceous glands: an autoradiographic study in subjects with acne vulgaris. AB - The interaction between spironolactone and dihydrotestosterone (DHT) receptors was evaluated with an autoradiographic technique. The inhibition of DHT receptors by spironolactone was found to be related to the decrease of tritiated DHT granules in the sebaceous glands of the treated site. 6 male patients affected by acne vulgaris entered the study. The acute study was performed by applying to 25 cm2 of the back a cream containing 5% spironolactone under occlusive dressing. The dosage of spironolactone applied was 4 mg/cm2 for 48 h. The long-term study was performed by applying the same amount to the entire back, without occlusion, twice daily for 1 month. Skin biopsies were taken at the end of the treatment, incubated with tritiated DHT and processed for autoradiography. Both the acute and the long-term study revealed a decrease of the autoradiographic granules in the treated site. This effect is related to the binding of spironolactone with dihydrotestosterone receptors in the sebaceous glands. Our study demonstrates that 5% topical spironolactone cream acts as an antiandrogen in human sebaceous glands, competing with DHT receptors and producing a decrease of labelled DHT. At the concentrations used the effect has been only local. No side-effects were recorded during both studies. PMID- 2972663 TI - Identification of proteins in retinas and IPM from eyes with retinitis pigmentosa. AB - Opsin, the alpha-subunit of transducin, S-antigen, interphotoreceptor retinoid binding protein (IRBP) and cathepsin D were assessed in autopsy eyes from patients with retinitis pigmentosa (RP) and normal autopsy eyes. Immunochemical methods were used to determine the presence of these proteins on Western blots of retinal homogenates from five RP donors and on blots of interphotoreceptor matrix (IPM) preparations from six other RP eyes. The amounts of immunoreactive opsin, S antigen, alpha-transducin, and IRBP appeared below normal in retinas from RP eyes. All six IPM samples from patients with advanced RP had reduced amounts of S antigen and no detectable IRBP or transducin. Cathepsin D (an RPE protein) was present in IPM or RP eyes in amounts comparable to that in IPMs from normal eyes. Small amounts of cathepsin D were also detected in retinas from both normal and RP eyes. These studies show that proteins specific to the photoreceptor-pigment epithelium complex in normal eyes can be detected in autopsy eyes from patients with RP and suggest that the observed reductions in photoreceptor-specific proteins occur as a consequence of photoreceptor loss. PMID- 2972664 TI - Pulmonary tuberous sclerosis responds to progesterone treatment. PMID- 2972665 TI - [Polyvinylpyrrolidone dermatoses. Clinical aspects and ultrastructural morphology]. AB - High-molecular polyvinylpyrrolidone (PVP) is used in order to induce a depot of subcutaneously or intramuscularly applied drugs. Deposits of PVP in skin plaques persisting for years and distant from the injection site have been demonstrated by the use of a modified fixation and embedding method for electron microscopy. The storage of PVP did not only occur in macrophages but also in other dermal cells whose pinocytotic activity seems to have been underestimated so far. PVP was found in mucinous and serous cells of sweat glands, in myoepithelial cells, endothelial cells, mast cells, and in the perineurium as well as in Schwann cells. PMID- 2972666 TI - [Pemphigus herpetiformis following D-penicillamine in a patient with HLA B8]. AB - A 57-year-old patient with rheumatoid arthritis, who was being treated with D penicillamine, developed a bullous eruption 2 months after the onset of treatment. On the basis of the clinical, histological and immunofluorescent findings, the diagnosis of herpetiform pemphigus was made. The eruptions disappeared within 2 weeks after the discontinuation of D-penicillamine treatment. During the 16-month follow-up period there was no recurrence. It is noteworthy that a patch test with potassium iodide was positive and that testing for HLA antigens revealed HLA B8, A1, A2, and BW39. PMID- 2972668 TI - Radiologic diagnosis. Mild cardiomegaly with left atrial enlargement. Moveable caudal esophageal or gastric mass. PMID- 2972667 TI - The multi-dose clinical tolerance and pharmacokinetics of the combined radiosensitizers, Ro 03-8799 (pimonidazole) and SR 2508 (etanidazole). AB - The hypoxic cell radiosensitizers Ro 03-8799 (pimonidazole) and SR 2508 (etanidazole) have differing physico-chemical properties and clinical toxicities. The former is basic, lipophilic and produces an acute but transient central nervous system syndrome; the latter is neutral, hydrophilic and causes cumulative peripheral neuropathy. We therefore investigated the possibility of combining these agents to achieve additive radiosensitization with no enhancement of toxicity, as demonstrated in a rodent tumor model. Following a single dose study which showed a lack of interaction with respect to both toxicity and pharmacokinetics, twenty-one patients have now completed simultaneous drug administration on an escalating, multiple dose schedule. There has been no adverse acute interaction up to 0.75 g/m2 Ro 03-8799 with 2 g/m2 SR 2508 for 15 doses. At this dose-level, however, all patients experienced peripheral neuropathy. There was no adverse pharmacokinetic interaction, or perturbation of plasma pharmacokinetics between initial and final infusions. Tumor concentrations were determined in 48 biopsy samples 0-60 min after administration. Mean values normalized to a dose of 0.75 g/m2 Ro 03-8799 plus 2 g/m2 SR 2508 were 33 micrograms/g Ro 03-8799 and 74 micrograms/g SR 2508. These would be expected to produce a single-dose sensitizer enhancement ratio of around 1.5. The combination is predicted to be around 6.8 times more active than misonidazole, and superior to any single agent tested to date. The current schedules are reaching the limits of clinical tolerance, and an attempt is now being made to define the optimal regimen for use in a randomized clinical trial of the combination. PMID- 2972669 TI - Hydrogen bromide adduct of neocarzinostatin chromophore: one of the stable derivatives of native neocarzinostatin chromophore. PMID- 2972670 TI - In-vitro induction of resistance in coagulase-negative staphylococci to vancomycin and teicoplanin. AB - Twenty strains of coagulase-negative staphylococci (18 strains of Staphylococcus haemolyticus and two of S. epidermidis) were serially subcultured in broth media containing subinhibitory concentrations (half the MIC) of either vancomycin or teicoplanin. The MIC of the antibiotic was again measured after five passages in antibiotic-containing broth. The organisms were then subcultured in broth containing antibiotic concentrations half of the new MIC value. The experiment was terminated after 25 passages. Only one strain developed a four-fold increase in vancomycin MIC. On the other hand, 16 strains developed four-fold MIC increases to teicoplanin. PMID- 2972671 TI - Activity of A-56268 (TE-031), a new macrolide, against Toxoplasma gondii in mice. AB - The activity of A-56268 (TE-031), a new macrolide, was tested in a murine model of acute toxoplasmosis. All control animals died in 8 +/- 1 days, while all mice treated with nine daily doses of A-56268 at 300 mg/kg, administered by gavage, survived. Moreover, 41.6% of the surviving mice were free from cerebral infection with Toxoplasma gondii, as assessed by brain subpassage. A-56268 is active against T. gondii in vivo, but further studies are needed to determine its usefulness in the treatment of human toxoplasmosis. PMID- 2972673 TI - Periodic weight support effects on rat soleus fibers after hindlimb suspension. AB - The morphological and histochemical properties of the rat soleus were studied after 1 wk of hindlimb suspension, one model that removes the weight-bearing function of the hindlimbs. To examine the effectiveness of weight support activity in maintaining soleus mass, fiber size, and succinate dehydrogenase (SDH) activity, the hindlimbs of adult male Sprague-Dawley rats were suspended (HS) and half of these rats were walked on a treadmill for 40 min/day (10 min every 6 h) at 5 m/min and a 19 degree grade (HS-WS). Significant reductions in soleus mass and fiber size were found after 1 wk of HS. Weight support activity decreased the atrophic response by approximately 50%. In the alkaline myofibrillar adenosine triphosphatase (ATPase) dark-staining fibers, SDH activity was higher in the HS than control rats, whereas it was similar to control in the HS-WS rats. Total SDH activity (SDH activity X cross-sectional area) in fibers staining lightly for ATPase in HS and HS-WS rats was lower than in control rats, whereas in the darkly stained ATPase fibers it was similar among the three groups. No changes were observed in fiber type percentages after 1 wk of HS or HS WS. The results suggest that short-duration, daily weight support activity can ameliorate, but not prevent, soleus atrophy induced by HS. Furthermore, fiber cross-sectional area is more responsive to periodic weight support in dark than light ATPase fibers. These results also demonstrate that muscle fiber atrophy need not be associated with a loss in SDH activity. PMID- 2972674 TI - Central inspiratory influence on abdominal expiratory nerve activity. AB - Our purpose was to determine whether the intensity of abdominal expiratory nerve discharge is conditioned by the intensity of the preceding inspiratory phrenic discharge, independent of mechanical and chemical afferent influences. In decerebrate, paralyzed, vagotomized cats with bilateral pneumothoraxes, we recorded phrenic and abdominal (cranial iliohypogastric nerve, L1) nerve activities at hyperoxic normocapnia. We reduced the duration and intensity (i.e., integrated peak height) of phrenic nerve discharge for single cycles by stimulating the cut central end of the superior laryngeal nerve (SLN) during the central inspiratory phase (75 microA, 20-50 Hz, 0.2-ms pulse). Premature termination of inspiration consistently reduced expiratory duration (TE) and abdominal expiratory nerve activity (area of integrated neurogram), but the average reduction in TE was much less than the reduction in abdominal nerve activity (14 vs. 51%). Stimulation of the cut central end of the vagus nerve yielded similar results, as did spontaneous premature terminations of inspiration, which we observed in one cat. SLN stimulation during hyperoxic hypercapnia resulted in more variable responses, and higher stimulation frequencies were usually required to abort inspiration. SLN (or vagal) stimulation during expiration consistently increased abdominal expiratory nerve activity. We speculate that this facilitatory response is gated during inspiration, thereby allowing the inspiratory conditioning effect on the subsequent expiration to be expressed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972672 TI - Changes in fiber composition of soleus muscle during rat hindlimb suspension. AB - Chronic reduction of gravitational load in the rear limbs of rats to simulate the influence of near-zero gravity in skeletal muscles has been shown previously to elicit atrophy in the soleus muscle. Use of this model by the present investigation indicates that soleus atrophy was characterized by a decline in the number of fibers in groups that contained the slow isoenzyme of myosin and which were classified as type I from intensity of staining to myofibrillar actomyosin adenosinetriphosphatase (ATPase) and to NADH tetrazolium reductase. Furthermore total fiber number was not changed, whereas fibers containing the intermediate isoenzyme and those classified as type IIa increased. There results could be explained by either a change in the composition within existing fibers or a simultaneous loss of slow fibers and de novo synthesis of intermediate and fast fibers. Evidence for transformation included an absence of embryonic or neonatal myosin in muscles from suspended rats and the constant fiber number that was unchanged by 4 wk of suspension. Furthermore although fiber areas of both groups of type I and IIa fibers declined during suspension, variability of the fiber areas within each group did not increase. PMID- 2972676 TI - Factor structure of the graduate record examinations general test in handicapped and nonhandicapped groups. PMID- 2972675 TI - Atrial natriuretic peptide lowers pulmonary arterial pressure in hypoxia-adapted rats. AB - To test the hypothesis that atrial natriuretic peptide (ANP) has a direct vasodilator effect on the pulmonary vasculature that is enhanced in hypoxia induced pulmonary hypertension in the rat, we determined the effects of ANP on mean pulmonary (MPAP) and systemic arterial pressure (MSAP) in intact conscious Sprague-Dawley rats exposed to 10% O2 or room air for 4 wk. Catheters were placed in the pulmonary artery through the right jugular vein by means of a closed-chest technique. MPAP and MSAP were monitored before and after intravenous injections of graded doses of ANP. ANP produced dose-related decreases in MPAP that were greater in the hypoxic group than in air controls. There were no significant between-group differences in the systemic depressor responses to ANP or in the ANP-induced reduction in cardiac output. ANP lowered MPAP significantly in isolated perfused lungs from both hypoxia-adapted and air control rats, and this effect was significantly greater in the hypoxic than the air control lungs. These data indicate that ANP lowers pulmonary arterial pressure in rats with hypoxia induced pulmonary hypertension, mainly by a direct vasodilator effect on the pulmonary vasculature. PMID- 2972677 TI - Fitness to drive and the general practitioner. AB - This study investigates general practitioners' (GPs') knowledge and opinions on fitness to drive after stroke. Questionnaires were sent to 650 GPs. Despite a low response rate (24%) results suggest that most GPs rarely see stroke patients regarding driving fitness. There appears to be some confusion over terms used in the handbook issued to medical practitioners. Only a quarter of GPs knew of any driving assessment facilities and very few had ever referred a patient to a clinical psychologist for cognitive assessment. Almost all responding GPs (98%) wanted the opportunity to refer patients for assessment based on research findings. The current approach to assessment of driving fitness after stroke appears inconsistent. It is hoped that new assessment methods being developed will help to improve on the present system. PMID- 2972678 TI - Individual and collective initiatives in disablement. AB - After setting a historical background for health care activities this paper relates the development of self-care to changes that have taken place. Relatively recent technological possibilities in medicine have altered the tensions in regard to self-care, highlighting demands for greater knowledge and the search for means of coming to terms with the existential challenge of chronic illness or disablement. The characteristics of self-help groups are discussed briefly, and problems are noted in relation to condition-specific orientation and the emphasis on personal identity and personal responsibility. Various contradictions in attitudes to self-help are also reviewed, notably those associated with the almost opposing traditions of mutual aid and individual self-sufficiency, in moral as well as economic dimensions. The ambiguous liberation provided by self care and self-help has to be set against political attempts to promote the notion that we can all look after ourselves. Among other things, this rhetoric serves to conceal the real challenges for improvements in community care. PMID- 2972679 TI - Update: September 1988. AIDS and adolescents: the need for prevention. PMID- 2972680 TI - Patient autonomy: one man's story. PMID- 2972681 TI - Advance directives in Arkansas. PMID- 2972682 TI - Remedy for frivolous malpractice actions. PMID- 2972683 TI - University of Arkansas for Medical Sciences medical student awards. PMID- 2972684 TI - James Henry Southall, M.D. 1841-1901. PMID- 2972686 TI - Dilemmograms. PMID- 2972685 TI - Update: October 1988 physician survey about AIDS. PMID- 2972687 TI - Migraine headaches in children. PMID- 2972688 TI - A team effort. PMID- 2972689 TI - Early identification and habilitation services. PMID- 2972690 TI - Treatment of Stage A1 prostate cancer. PMID- 2972691 TI - Low plasma immunoreactive beta-endorphin levels in autism. PMID- 2972692 TI - Autistic social dysfunction and Down syndrome. PMID- 2972693 TI - Absence of antibodies to human immunodeficiency virus (HIV) in health workers after hepatitis B vaccination. PMID- 2972694 TI - Dextransucrase secretion in Leuconostoc mesenteroides depends on the presence of a transmembrane proton gradient. AB - The relationship between proton motive force and the secretion of dextransucrase in Leuconostoc mesenteroides was investigated. L. mesenteroides was able to maintain a constant proton motive force of -130 mV when grown in batch fermentors at pH values 5.8 to 7.0. The contribution of the membrane potential and the transmembrane pH gradient varied depending on the pH of the growth medium. The differential rate of dextransucrase secretion was relatively constant at 1,040 delta mU/delta mg (dry weight) when cells were grown at pH 6.0 to 6.7. Over this pH range, the internal pH was alkaline with respect to the external pH. When cells were grown at alkaline pH values, dextransucrase secretion was severely inhibited. This inhibition was accompanied by an inversion of the pH gradient as the internal pH became more acidic than the external pH. Addition of nigericin to cells at alkaline pH partially dissipated the inverted pH gradient and produced a fourfold stimulation of dextransucrase secretion. Treatment of cells with the lipophilic cation methyltriphenylphosphonium had no effect on the rate of dextransucrase secretion at pH 5.5 but inhibited secretion by 95% at pH 7.0. The reduced rate of secretion correlated with the dissipation of the proton motive force by this compound. Values of proton motive force greater than -90 mV were required for maximal rates of dextransucrase secretion. The results of this study indicate that dextransucrase secretion in L. mesenteroides is dependent on the presence of a proton gradient across the cytoplasmic membrane that is directed into the cell. PMID- 2972696 TI - Psychiatric morbidity associated with early clinical diagnosis of Huntington disease in a predictive testing program. AB - Predictive testing for Huntington disease is now possible by using DNA markers close to the mutant gene. Such tests result in an increased or decreased risk estimate as to whether the person tested has inherited the mutant gene. Persons who already manifest subtle signs of the disease but see themselves as presymptomatic will also present to the preclinical testing programs. The authors report a severe psychiatric reaction to a positive clinical diagnosis of Huntington disease in a woman enrolled in a pilot preclinical program who had a history of psychiatric illness and hospitalizations. She is the only one of 60 persons currently enrolled in the program who has had a serious crisis. The authors demonstrate the need for readily available and ongoing counseling services for all persons participating in such a program. PMID- 2972695 TI - Analysis of transcription termination signals in the nin region of bacteriophage lambda: the roc deletion. AB - Deletions in the region, nin, between the P and Q genes of phage lambda remove a portion of the phage genome that includes signals for termination of transcription. These deletions were selected because they permit growth of lambda derivatives defective in the N-mediated transcription antitermination system; i.e., the deletions confer N independence (nin). Thus nin phages (e.g., lambda nin5) grow in most Escherichia coli nus mutants. The nus genes encode functions necessary for N action. We report the isolation of a deletion in the nin region delta roc that confers a partially N-independent phenotype; lambda derivatives with delta roc can grow under normally nonpermissive conditions, 32 degrees C, in a host with the rpoB-nusC60 mutation. The roc deletion also partially suppresses the inhibitory effects of other nus mutations at higher temperatures. Delta roc, which extends from base pairs 41883 to 43825, overlaps the nin5 deletion, which extend from base pairs 40501 to 43306. Unlike the nin5 deletion, the sequences deleted by delta roc do not include a stem-loop structure, tR2, previously shown to have terminator activity. Using promoter and terminator testor vectors, we demonstrate that a 2,400-base-pair fragment that includes the wild-type roc region but excludes tR2 has terminator activity. Thus, delta roc permits a functional division of the transcription termination signals in the nin region. PMID- 2972697 TI - Elementary steps of the actin-activated ATPase reaction of cardiac muscle myosin subfragment-1. AB - The rates of the elementary steps of the actomyosin ATPase reaction were measured using the myosin subfragment-1 of porcine left ventricular muscle. The results could be explained only by the two-route mechanism for actomyosin ATPase (Inoue, Shigekawa, & Tonomura (1973) J. Biochem. 74, 923-934), in which ATP is hydrolyzed via routes with or without accompanying dissociation of actomyosin. The dependence on the F-actin concentration of the rate of the acto-S-1 ATPase reaction in the steady state was measured in 5 mM KCl at 20 degrees C. The maximal rate, Vmax, and the dissociation constant for F-actin of the ATPase, Kd, were 3.0 s-1 and 2.2 mg/ml, respectively. The Kd value was almost the same as that determined from the extent of binding of S-1 with F-actin during the ATPase reaction. The rate of recombination of the S-1-phosphate-ADP complex, S-1ADPP, with F-actin, vr, was lower than that of the ATPase reaction in the steady state. Thus, ATP is mainly hydrolyzed without accompanying dissociation of acto-S-1 into S-1ADPP and F-actin. In the cardiac acto-S-1 ATPase reaction, the rate of the ATPase reaction in the steady state and that of recombination of S-1ADPP with F actin were about 1/5 those of the skeletal acto-S-1 ATPase reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972699 TI - p-nitrophenyl penta-N-acetyl-beta-chitopentaoside as a novel synthetic substrate for the colorimetric assay of lysozyme. AB - p-Nitrophenyl beta-glycosides of N-acetylchitooligosaccharides (PNP-(GlcNAc)n n = 3-5) were examined as substrates for lysozyme [EC 3.2.1.17]. The enzyme released predominantly p-nitrophenyl N-acetyl-beta-D-glucosaminide (PNP-GlcNAc) from each substrate. Furthermore, the initial rate of PNP-GlcNAc formation in lysozyme catalyzed hydrolysis of p-nitrophenyl penta-N-acetyl-beta-chitopentaoside (PNP (GlcNAc)5) was about 350 and 25 times faster than those of p-nitrophenyl tri-N acetyl-beta-chitotrioside (PNP-(GlcNAc)3) and p-nitrophenyl tetra-N-acetyl-beta chitotetraoside (PNP-(GlcNAc)4), respectively. From these results, a new colorimetric assay method of lysozyme using PNP-(GlcNAc)5 as a substrate was developed on the basis of the determination of p-nitrophenol liberated from the substrate by lysozyme through a coupled reaction involving beta-N acetylhexosaminidase (NAHase). The assay system gave a linear dose-response curve in the range of 2-120 micrograms of lysozyme in a 15-60 min incubation. The present assay was not significantly influenced by the ionic strength of the medium and was reproducible. This method using PNP-(GlcNAc)5 as a substrate was shown to be useful for lysozyme assay. PMID- 2972698 TI - Biochemical characteristics of the Mr 52,000 component of Akazara scallop troponin. AB - Some biochemical properties of the Mr 52,000 component of Akazara scallop striated adductor troponin, which had been tentatively identified as troponin-I, were compared with those of rabbit troponin-I. Both the Mr 52,000 component and rabbit troponin-I together with rabbit tropomyosin inhibited the Mg-ATPase activity of rabbit reconstituted actomyosin to 1/10 of the original activity. The inhibition was neutralized by the addition of Akazara scallop and rabbit troponin C or Patinopecten scallop calmodulin. The Mr 52,000 component and rabbit troponin I were insoluble below 0.15 M KCl, but were solubilized by complexing with an equimolar amount of troponin-C or calmodulin. On alkaline urea-polyacrylamide gel electrophoresis, the Mr 52,000 component as well as rabbit troponin-I was found to form a stable complex with troponin-C or calmodulin in the presence of Ca2+. PMID- 2972700 TI - Structural aspects of skeletal muscle F-actin as studied by tryptic digestion: evidence for a second nucleotide interacting site. AB - Trypsin was used as a probe of F-actin conformation. F-actin is known to be refractory to proteolysis [Jacobson, G.R. and Rosenbusch, J.P. (1976) Proc. Natl. Acad. Sci. U.S. 73, 2742-2746]. However, here it was found that F-actin could also be digested by trypsin to a 33-kDa fragment (like G-actin) when free MgADP is present in the medium. The amounts of degradation of F-actin depended on the ADP concentration; saturation occurred at about 0.5 mM. Elimination of divalent cations from the medium completely suppressed the effect of ADP on the digestion of F-actin. Other nucleotides were also examined. The effect decreased in the order ADP greater than ATP much greater than IDP greater than GDP = UDP. Adenine, adenosine, AMP, and PPi had no effect at all. epsilon-ADP had the effect, and its fluorescence was changed on the addition of F-actin. The intrinsic tryptophan fluorescence spectrum of F-actin was ADP-dependent. These results suggest the presence of a second nucleotide interacting site on actin and that ADP interaction at this site induces conformational changes in monomeric actin molecule in F-actin filaments. PMID- 2972701 TI - Characterization of carbethoxylated actin. AB - The carbethoxylation of histidine residues in G-actin impairs actin polymerization. The histidine residue essential for polymerization was identified as histidine-40 [Hegyi, G., Premecz, G., Sain, B., & Muhlrad, A. (1974) Eur. J. Biochem. 44, 7-12]. Non-polymerizable actin was separated from the polymerizable fraction after partial carbethoxylation. The non-polymerizable actin recovered the ability to polymerize following addition of phalloidin. Taking into account the evidence that phalloidin does not bind to G-actin in the absence of salt, the results indicate that the actin monomer undergoes a conformational change and subsequently binds phalloidin before polymerization. The resulting polymers activated S1 ATPase activity as effectively as control F-actin. In the presence of tropomyosin and troponin, a strong inhibition of actin-activated ATPase activity was observed in the absence of Ca2+, although no inhibition was observed in the presence of Ca2+. These results indicate that His-40 is not directly involved in a myosin binding site nor in a tropomyosin-troponin binding site. PMID- 2972702 TI - Dynein ATPases as microtubule motors. PMID- 2972703 TI - The stereochemical course of phospho group transfer catalyzed by rat liver 6 phosphofructo-2-kinase. AB - Adenosine [gamma-(S)-16O,17O,18O]triphosphate was used as substrate in the phosphokinase reaction catalyzed by 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase. The product D-fructose 2,6-[2-16O,17O,18O]bisphosphate was then used as substrate in the alkaline phosphatase-mediated transfer of the phospho groups to (S)-butane-1,3-diol, where the configuration at phosphorus has been determined. Although there was approximately equal transfer by alkaline phosphatase of the labeled 2- and the unlabeled 6-phospho groups, the subsequent assignment of configuration of the chiral phospho group from the 2-position was unambiguous. It was found that 6-phosphofructo-2-kinase proceeds by a pathway that results in net inversion of the configuration at phosphorus. The simplest interpretation of this result is that the phospho group is transferred directly between substrates in a ternary complex by an "in-line" mechanism not involving a phosphoenzyme intermediate. This conclusion is consistent with the fact that the enzyme cannot be labeled by [gamma-32P]ATP and with steady-state kinetic data that suggest an ordered sequential mechanism. This finding also indicates that the adenine nucleotide and sugar phosphate isotope exchange reactions catalyzed by the enzyme are not relevant to the normal catalytic pathway of the kinase. PMID- 2972704 TI - Secretion of atrial natriuretic factor-(1-98) by primary cardiac myocytes. AB - Previous studies have demonstrated that primary cultures of cardiac myocytes maintained in a complete serum-free medium contain a precursor to atrial natriuretic factor (ANF-(1-126]. The cultured cells secrete this precursor unless maintained in the presence of glucocorticoids wherein the known circulating form derived from the C-terminal of ANF (ANF-(99-126] is secreted. The present study was designed to determine the fate of the N-terminal region of the ANF precursor during secretion from myocytes maintained in glucocorticoids. A radioimmunoassay (RIA) was developed using synthetic ANF-(1-16); the antiserum demonstrated cross reactivity toward ANF-(1-126) and ANF-(1-98)-like peptides but did not cross react with ANF-(99-126). Coupling this RIA with an ANF-(99-126)-specific RIA and reversed phase, size exclusion, and ion exchange high performance liquid chromatography (HPLC), it was shown that primary cultures of atrial myocytes maintained in dexamethasone contained ANF-(1-126) and secreted ANF-(99-126) and a peptide that was chromatographically indistinguishable from ANF-(1-98). Isolated perfused rat hearts were also shown by RIA and HPLC to secrete similar peptides. The primary cells were labeled with [35S]methionine, and the secreted N-terminal ANF-related material was immunoprecipitated with the ANF-(1-16) antiserum. HPLC, tryptic peptide mapping, and radiosequencing demonstrated that this peptide possessed an N-terminal structure identical to that of ANF-(1-126). When the cells were labeled with [3H] leucine and the secreted N-terminal ANF-related material was immunoprecipitated and analyzed by tryptic mapping, it was shown to possess labeled tryptic peptides consistent with the structure of ANF-(1-98). Tryptic mapping of [3H]arginine-labeled N-terminal ANF-related material demonstrated the presence of all peptides consistent with the ANF-(1-98) structure, including ANF-(92-98). These studies demonstrate that primary atrial myocytes contain ANF-(1-126) and in the presence of dexamethasone secrete both ANF-(1-98) and ANF-(99-126), the two major circulating forms of the hormone. PMID- 2972705 TI - Expression of human cation-independent mannose 6-phosphate receptor cDNA in receptor-negative mouse P388D1 cells following gene transfer. AB - We recently reported the cDNA cloning, sequence, and expression of the human cation-independent mannose 6-phosphate receptor (hCI-MPR) (Oshima, A., Nolan, C. M., Kyle, J. W., Grubb, J. H., and Sly, W. S. (1988) J. Biol. Chem. 263, 2553 2562). The sequence of the hCI-MPR was virtually identical to that of the human insulin-like growth factor II receptor cDNA (Morgan, D. O., Edman, J. C., Standring, D. N., Fried, V. A., Smith, M. C., Roth, R. A., and Rutter, W. J. (1987) Nature 329, 301-307). To test the role of the putative bifunctional receptor in intracellular sorting of acid hydrolases, we studied its effect on lysosomal enzyme transport following gene transfer to receptor-negative cells. Receptor-negative mouse P388D1 cells were transfected with a cDNA construct containing the entire coding sequence of hCI-MPR under the control of the mouse metallothionine I promoter. Stable transformants were isolated and characterized. The expressed hCI-MPR was localized in membranes including the plasma membrane, bound mannose 6-phosphate containing ligands, and mediated endocytosis which could be specifically blocked by mannose 6-phosphate. We next measured the effect of the expressed hCI-MPR on intracellular and secreted acid hydrolases. The intracellular activity of the lysosomal marker enzymes beta-glucuronidase and beta-hexosaminidase increased up to 2-fold following transformation. In addition, expression of the receptor greatly reduced the fraction of acid hydrolases secreted. These phenotypic changes in the transformed cell lines support the proposed role of the cation-independent mannose 6-phosphate receptor in intracellular sorting and targeting of lysosomal enzymes. PMID- 2972706 TI - The structural basis for insulin-like growth factor I receptor high affinity binding. AB - We have recently identified high and low affinity insulin-like growth factor I (IGF I) binding sites in solubilized human placental membranes and purified the high affinity IGF I receptor by IGF I affinity chromatography (Tollefsen, S. E., Thompson, K., and Petersen, D. J. (1987) J. Biol. Chem. 262, 16461-16469). To define the structural basis for high affinity IGF I binding, we have examined the effect of disulfide bond reduction on the binding parameters of the high affinity IGF I receptor. We find that the disulfide bonds linking the two alpha beta dimers of the IGF I receptor heterotetramer are reduced by incubation at pH 8.75 with 2 mM dithiothreitol (DTT) for 5 min at room temperature. Gel filtration chromatography on a Superose 12 fast protein liquid chromatography column indicates that the alpha beta dimers do not remain associated by noncovalent interactions after reduction. Scatchard plots of IGF I binding to the IGF I receptor incubated at pH 8.75 with or without DTT indicate that the IGF I receptor alpha beta dimers have a 6.1 +/- 1.6 (mean +/- S.D.) times lower affinity than the heterotetramer for IGF I. The total binding capacity of the IGF I receptor treated with DTT is 1.6 +/- 0.3 (mean +/- S.D.) times higher than that of an equal amount of receptor treated without DTT. These results are consistent with a model in which the heterotetramer binds a single IGF I molecule with high affinity, whereas each of the two alpha beta dimers binds an IGF I molecule with lower affinity after dissociation. We conclude that association of two alpha beta dimers is required for formation of an IGF I receptor with high affinity for its ligand. PMID- 2972707 TI - Biological activity and receptor binding of human prointerleukin-1 beta and subpeptides. AB - We report here that the human interleukin-1 beta precursor (proIL-1 beta) protein as well as several interleukin-1 beta (IL-1 beta) subpeptides bind cellular receptors specifically and exhibit biological activity by stimulating proliferation of helper T-cells. IL-1 beta polypeptides have been synthesized by in vitro translation of mRNAs transcribed from plasmid vectors containing the bacteriophage SP6 promoter joined to the complete IL-1 beta cDNA or to deletion constructs. The quantity of IL-1 beta in vitro translation products was increased significantly by replacing the cognate IL-1 beta untranslated leader sequence with a 37-nucleotide plant viral untranslated leader. Translation of chimeric mRNAs followed by direct bioactivity assay demonstrated that mature IL-1 beta (117-269), proIL-1 beta-(1-269), and peptide IL-1-(71-269) were all biologically active. Specific binding to cellular receptors was observed with these three IL-1 beta molecules; moreover, several peptides with minimal biological activity also bound receptor specifically. The biological activity and receptor binding properties of the IL-1 beta proteins reported here contrast with those described by Mosley et al. (Mosley, B., Urdal, D. L., Prickett, K. S., Larsen, A., Cosman, D., Conlon, P. J., Gillis, S., and Dower, S. K. (1987) J. Biol. Chem. 262, 2941 2944; Mosley, B., Dower, S. K., Gillis, S., and Cosman, D. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 4572-4576), who reported that proIL-1 beta-(1-269) had no biological activity and does not bind receptor. Our results indicate that proIL-1 beta is active at a relatively high concentration, and analysis of the proIL-1 beta-(1-269) and IL-1-(71-269) bioactivity data suggests a possible relationship with membrane-bound IL-1. PMID- 2972708 TI - Identification of cDNA clones encoding different domains of the basement membrane heparan sulfate proteoglycan. AB - We have used antibodies to the basement membrane proteoglycan to screen lambda gt11 expression vector libraries and have isolated two cDNA clones, termed BPG 5 and BPG 7, which encode different portions of the core protein of the heparan sulfate basement membrane proteoglycan. These clones hybridize to a single mRNA species of approximately 12 kilobases. Amino acid sequences obtained on peptides derived from protease digests of the core protein were found in the deduced sequence, confirming the identity of these clones. BPG 5 spanned 1986 base pairs and has an open reading frame of 662 amino acids. The amino acid sequence deduced from BPG 5 contains two cysteine-rich domains and two internally homologous domains lacking cysteine. The cysteine-rich domains show homology to the cysteine rich domains of the laminin chains. A globule-rod structure, similar to that of the short arms of the laminin chains, is proposed for this region of the proteoglycan. The other clone, BPG 7, is 2193 base pairs long and has an open reading frame of 731 amino acids. The deduced sequence contains eight internal repeats with 2 cysteine residues in each repeat. These repeats show homology to the neural-cell adhesion molecule N-CAM and the plasma alpha 1B-glycoprotein. Looping structures similar to these proteins and to other proteins of the immunoglobulin gene superfamily are proposed for this region of the proteoglycan. The sequence DSGEY was found four times in this domain and could be heparan sulfate attachment sites. PMID- 2972709 TI - Immunoprecipitation of interleukin-1 receptors from a mouse thymoma cell line, EL 4. AB - Receptors for the monokine, interleukin-1 (IL-1), have been successfully immunoprecipitated with a xenogeneic antiserum raised in our laboratories. Receptors solubilized from mouse cell membranes as well as nascent chains of molecules that could bind IL-1 were immunoprecipitated. Receptor complexes were identified on mouse cell lines which express IL-1 receptors by affinity cross linkage of the radiolabeled ligands, IL-1-alpha or IL-1-beta. Soluble IL-1 or IL 1 nonspecifically associated with membranes of cells which do not express IL-1 receptors was not immunoprecipitated. It is apparent, thus, that antibodies present in the xenogeneic antiserum could specifically bind to the IL-1 receptor moiety within the complex. The major proportion of IL-1 receptor complexes that were reproducibly immunoprecipitated had a molecular weight of 97,000. Cell membrane associated receptors for the monokine, tumor necrosis factor, were not immunoprecipitated. These antibodies have contributed to the understanding of the role of IL-1 receptors in cytolytic effector T cell generation and should contribute further in the purification and characterization of the IL-1 receptor moiety, as well as in determining IL-1-mediated mechanisms of cellular activation. PMID- 2972710 TI - Interleukin-1 receptor-like proteins synthesized by translation, in vitro, of immunopurified polysomal messenger RNA. AB - Interleukin-1 (IL-1) receptors can be solubilized from murine cell surfaces and immunoprecipitated with a xenogeneic rat antiserum raised in this laboratory. We demonstrated first that this antiserum contains antibodies directed against IL-1 receptors. We have now successfully used this antiserum as a reagent to immunopurify polysomes along with their messenger RNA from a murine leukemic cell line known to express relatively high levels of IL-1 receptors. The immunoselected mRNA was translated into proteins in vitro. The translation products contained an IL-1 binding protein which could specifically bind to immobilized IL-1 but not to other immobilized ligands such as interleukin-2 or tumor necrosis factor-alpha. The translation products which bound to IL-1 could be acid-eluted from the immobilized ligand, and the proteins released could still specifically bind to IL-1 in a receptor-ligand binding reaction. The eluted IL-1 binding proteins, as well as soluble receptor-ligand complexes derived from them, could also be immunoprecipitated with the xenogeneic rat antiserum. The xenogeneic rat antiserum could, furthermore, immunoprecipitate the IL-1 binding proteins from the translated products before ligand was added. The residual translated products no longer interacted with IL-1. We conclude that our antiserum contains antibodies that recognize determinants expressed on the following proteins: on nascent chains of IL-1 binding proteins; on soluble translated IL-1 binding proteins; on soluble complexes of IL-1 binding proteins that had been cross-linked with IL-1 ligand; and on cell surface-associated IL-1 receptors. The translated and unprocessed IL-1 binding proteins have a molecular mass of approximately 52,000-56,000 daltons. PMID- 2972712 TI - Studies on the role of the phi X174 gene A protein in phi X viral strand synthesis. II. Effects of DNA replication of mutations in the 30-nucleotide icosahedral bacteriophage origin. AB - phi X174 viral strand circular DNA can be synthesized in vitro from phi X174 replicative form I (RFI) DNA in the presence of the phi X A protein, the Escherichia coli DNA polymerase III elongation system, the E. coli rep helicase, and the E. coli single-stranded DNA binding protein. M13mp9 or pBR322 RFI DNAs containing a 30-base pair sequence found at the phi X origin of replication supported phi X A protein synthesis as well as the phi X template, giving rise to a net molar excess of deoxynucleotide incorporation. In this paper, we show that mutations in positions 1-3 of the 30-nucleotide origin replicated at a lower efficiency than plasmids containing the wild-type origin, because of a deficiency in the reinitiation reaction. Mutations in positions 4-7, upstream of the phi X A protein cleavage site, failed to support replication because of their inability to support nicking. An origin containing a mutation at the residue to which the phi X A protein is covalently linked to the DNA was an active template that supported a net molar excess of incorporation. Mutations at the 3' end of the origin region, retaining only the first 21-25 nucleotides of the 30-base pair origin, failed to support replication because of impaired binding of the phi X A protein to the template and consequently poor nicking. A construct bearing the first 28 nucleotides of the origin supported wild-type replication, as did a plasmid containing a 28-mer origin with a point mutation at position 26, but this latter construct also appeared to be partially deficient in phi X A protein binding activity. These results are consistent with the presence of a phi X A protein binding domain at the 3' end of the origin. PMID- 2972711 TI - Studies on the role of the phi X174 gene A protein in phi X viral strand synthesis. I. Replication of DNA containing an alteration in position 1 of the 30 nucleotide icosahedral bacteriophage origin. AB - The influence of a C----G transversion at position 1 of the 30-base pair replication origin of bacteriophage phi X174 replicative form I DNA (phi X RFI) was examined in the RF----single-stranded circular DNA replication pathway catalyzed by the combined action of the purified phi X A protein, the Escherichia coli DNA polymerase III holoenzyme, rep helicase, and single-stranded DNA binding protein (Eisenberg, S., Scott, J.F., and Kornberg, A. (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 1594-1597; Reinberg, D., Zipursky, S.L., and Hurwitz, J. (1981) J. Biol. Chem. 256, 13143-13151). RFI DNA containing this transversion was cleaved to RFII by the phi X A protein as effectively as DNA containing the wild type origin. The altered duplex DNA, however, supported replication at a slower rate (3- to 4-fold) than the wild-type DNA due to a defect in the termination and reinitiation reactions catalyzed by the phi X A protein. This defect resulted in the accumulation of DNA products containing long single strands covalently joined to the mutant DNA. These single strands were susceptible to nuclease S1 and exonuclease VII attack. The defect in the template DNA containing C----G transversion was not corrected when this mutant origin was placed on the same strand with a wild-type origin. This double-origin DNA was also replicated poorly and led to the accumulation of large products, in contrast to the products formed with RFI DNA containing two wild-type 30-base pair replication origins on the same strand. PMID- 2972713 TI - Studies on the role of the phi X174 gene A protein in phi X174 viral strand synthesis. III. Replication of DNA containing two viral replication origins. AB - Supercoiled plasmid bearing two wild-type phi X origin sequences on the same strand supported the phi X A protein-dependent in vitro formation of two smaller single-stranded circles, the lengths of which were equivalent to the distance between the two origins. Additional double origin plasmids were utilized to determine whether origins defective in the initial nicking event (initiation) could support circularization (termination). In all cases tested, the presence of a mutant origin on the same strand with a wild-type origin affected the level of replication in a manner consistent with the previously determined activity of the mutant origin. When a functional mutant origin was present on the same strand as a wild-type origin, the efficiency of replication and the DNA products formed were almost identical to those of the plasmid containing two wild-type origins. Plasmid DNA bearing both a wild-type origin and a mutant origin that did not support phi X A protein binding or nicking activity, on the other hand, supported efficient DNA synthesis of only full-length circular products, indicating that the origin defective for initiation was incapable of supporting termination. In contrast, the presence of a wild-type origin and an origin that did bind the phi X A protein but was not cleaved resulted in a marked decrease in DNA synthesis along with the production of only full-length products. This suggests that the phi X A protein stalls when it encounters a sequence to which it can bind but cannot cleave. Replication of double origin plasmids containing one functional phi X origin on each strand of the supercoiled DNA was also examined. With such templates, synthesis from the wild-type origin predominated, indicating preferential cleavage of the intact origin sequence. Replication of such substrates also produced a number of aberrant structures, the properties of which suggested that interstrand exchange of the phi X A protein had occurred. PMID- 2972714 TI - Effect of phi X C protein on leading strand DNA synthesis in the phi X174 replication pathway. AB - The influence of the bacteriophage phi X174 (phi X) C protein on the replication of bacteriophage phi X174 DNA has been examined. This small viral protein, which is required for the packaging of phi X DNA into proheads, inhibits leading strand DNA synthesis. The inhibitory effect of the phi X C protein requires a DNA template bearing an intact 30-base pair (bp) phi X origin of DNA replication that is the target site recognized by the phi X A protein. Removal of nucleotides from the 3' end of this 30-bp conserved origin sequence prevents the inhibitory effects of the phi X C protein. Leading strand replication of supercoiled DNA substrates containing the wild-type phi X replication origin results in the production of single-stranded circular DNA as well as the formation of small amounts of multimeric and sigma structures. These aberrant products are formed when the termination and reinitiation steps of the replication pathway reactions are skipped as the replication fork moves through the origin sequence. Replication carried out in the presence of the phi X C protein leads to a marked decrease in these aberrant structures. While the exact mechanism of action of the phi X C protein is not clear, the results presented here suggest that the phi X C protein slows the movement of the replication fork through the 30-bp origin sequence, thereby increasing the fidelity of the termination and reinitiation reactions. In keeping with the requirement for the phi X C protein for efficient packaging of progeny phi X DNA into proheads, the phi X C protein-mediated inhibition of leading strand synthesis is reversed by the addition of proteins essential for phi X bacteriophage formation. Incubation of plasmid DNA substrates bearing mutant 30 base pair phi X origin sequences in the complete packaging system results in the in vitro packaging and production of infectious particles in a manner consistent with the replication activity of the origin under study. PMID- 2972715 TI - Structure of the murine complement factor H gene. AB - Factor H is a regulatory protein of the alternative pathway of complement activation comprised of 20 tandem repeating units of 60 amino acids each. A factor H cDNA clone was used to identify 17 genomic clones from a cosmid library. Four clones were selected for analysis of intron/exon junctions and 5' and 3' regions of the gene and for mapping of the exons. The factor H gene was found to be comprised of 22 exons. Each repeating unit is encoded by one exon, except the second repeat, which is coded by two exons; the leader sequence is encoded by a separate exon. The exons range in size from 77 to 210 base pairs (bp) and average 178 bp. They span a region of approximately 100 kilobases (kb) on chromosome 1. The leader sequence exon is 26 kb upstream of the first repeat exon, representing the largest intron. The other introns range in size from 86 bp to 12.9 kb, and the average intron size is 4.7 kb. Analysis of the genomic organization of the factor H gene has provided insight into the protein structure and will enable the construction of deletion mutants for functional studies. PMID- 2972716 TI - Molecular cloning of the cDNA which encodes beta-N-acetylhexosaminidase A from Dictyostelium discoideum. Complete amino acid sequence and homology with the human enzyme. AB - beta-N-Acetylhexosaminidase A (EC 3.2.1.52), the product of the nag A gene, is a lysosomal enzyme which is developmentally regulated in Dictyostelium discoideum. The enzyme plays a role during the slug stage of development in the maintenance of pseudoplasmodia of normal size. We used a homogeneous preparation of deglycosylated enzyme subunits to generate antibody. The antibody was suitable for screening a lambda gt11 cDNA expression library derived from the mRNA of late log stage axenic cells. We isolated seven positive clones. One of these contains the complete coding sequence of the protein. We also isolated a genomic clone which contains 800 base pairs of 5'-flanking sequence and 728 base pairs of coding sequence. Analysis of the sequences and of primer extension studies indicates an inferred transcript size of 1665 bases which closely matches the 1.8 kilobase mRNA size estimated by Northern blot analysis of poly(A+) mRNA from the organism. The sequence contains an open reading frame which encodes a protein of 59,787 kDa. This equals the apparent molecular weight in sodium dodecyl sulfate polyacrylamide gels of in vitro translated enzyme. The amino terminus of the purified enzyme appeared to be blocked, but internal peptide sequences were obtained by automated Edman degradation of gel-purified peptides generated by treatment of protein subunits with staphylococcal V-8 protease. These sequences are included in the inferred sequence. In addition to a typical signal sequence, the open reading frame encodes a second candidate transmembrane region, a serine rich region, and four potential N-glycosylation sites. These are discussed with regard to the localization and processing of the enzyme during its biogenesis. Beginning at amino acid 100 of the Dictyostelium enzyme sequence, 36% of its amino acids are identical to the corresponding sequence of the beta chain, and 33% are identical with those of the alpha chain of human beta-N acetylhexosaminidase. This is strong evidence that the Dictyostelium enzyme is homologous to the alpha and beta chains of the human enzyme. PMID- 2972718 TI - Structure of D-prephenyllactate. A carboxycyclohexadienyl metabolite from Neurospora crassa. AB - A novel natural product structurally related to prephenate and arogenate was isolated from a mutant of Neurospora crassa. This D-beta-(1-carboxy-4-hydroxy-2,5 cyclohexadiene-1-yl)-lactic acid is herein given the trivial name of D prephenyllactate. The new metabolite is even more acid labile than is prephenate and is quantitatively converted to phenyllactate at mildly acidic pH. The structure characterization of prephenyllactate was performed using spectroscopic techniques (ultraviolet, 1H NMR, 13C NMR, two-dimensional heteronuclear experiments and mass spectrometry). Circular dichroism proved conclusively the R configuration of the asymmetric carbon at C-8 of prephenyllactate. Enzymatic utilization of prephenyllactate by cyclohexadienyl dehydratase and by cyclohexadienyl dehydrogenase from Klebsiella pneumoniae was demonstrated. PMID- 2972717 TI - Isolation and characterization of microplasminogen. A low molecular weight form of plasminogen. AB - A functionally active human microplasminogen without kringle structures was produced by incubation of plasminogen with urokinase-free plasmin at an alkaline pH. The microplasminogen was purified by affinity chromatography on lysine- and soybean trypsin inhibitor-Sepharose and by chromofocusing. Human plasminogen is specifically cleaved at Arg529-Lys530 by plasmin to form microplasminogen, which consists of a single polypeptide of 261 residues from the COOH-terminal portion of native plasminogen. It has an Mr of 28,617, calculated from the sequence, which is consistent with the molecular weight determined by sodium dodecyl sulfate gel electrophoresis. Microplasminogen is a slightly basic protein and is eluted from a chromofocusing column at pH 8.3. It can be activated by urokinase and streptokinase to a catalytically active microplasmin. The specific amidolytic activity of microplasmin is about three times higher than Lys77-plasmin on a weight basis and is about the same on a molar basis. The activation of microplasminogen by streptokinase is slower than that of either Glu-plasminogen or Lys77-plasminogen. On the other hand, the activation of microplasminogen by urokinase is faster than that of either of the latter. The Arg560-Val561 bond is cleaved during activation of both microplasminogen and native plasminogen. PMID- 2972719 TI - Phosphofructokinase from bumblebee flight muscle. Molecular and catalytic properties and role of the enzyme in regulation of the fructose 6 phosphate/fructose 1,6-bisphosphate cycle. AB - Phosphofructokinase from the flight muscle of bumblebee was purified to homogeneity and its molecular and catalytic properties are presented. The kinetic behavior studies at pH 8.0 are consistent with random or compulsory-order ternary complex. At pH 7.4 the enzyme displays regulatory behavior with respect to both substrates, cooperativity toward fructose 6-phosphate, and inhibition by high concentration of ATP. Determinations of glycolytic intermediates in the flight muscle of insects exposed to low and normal temperatures showed statistically significant increases in the concentrations of AMP, fructose 2,6-bisphosphate, and glucose 6-phosphate during flight at 25 degrees C or rest at 5 degrees C. Measuring the activity of phosphofructokinase and fructose 1,6-bisphosphatase at 25 and 7.5 degrees C, in the presence of physiological concentrations of substrates and key effectors found in the muscle of bumblebee kept under different environmental temperatures and activity levels, suggests that the temperature dependence of fructose 6-phosphate/fructose 1,6-bisphosphate cycling may be regulated by fluctuation of fructose 2,6-bisphosphate concentration and changes in the affinity of both enzymes for substrates and effectors. Moreover, in the presence of in vivo concentrations of substrates, phosphofructokinase is inactive in the absence of fructose 2,6-bisphosphate. PMID- 2972720 TI - Pre-steady-state phosphorylation of the human red cell Ca2+-ATPase. AB - The pre-steady-state kinetics of phosphorylation of the Ca2+-ATPase by ATP was studied at 37 degrees C and in intact red cell membranes to approach physiological conditions. ATP and Ca2+ activate with K0.5 of 4.9 and 26.4 microM, respectively. Preincubation with Ca2+ did not change the K0.5 for ATP. Preincubation with ATP did not alter the initial velocity of phosphorylation suggesting that binding of ATP was not rate-limiting. Mg2+ added at the start of the reaction increased the initial rate of phosphorylation from 4 to 8 pmol/mg/s. With 30 microM Ca2+, the K0.5 for Mg2+ was 60 microM. Mg2+ and Ca2+ added together beforehand accelerated phosphorylation to 70 pmol/mg/s. Phosphorylation of calmodulin-bound membranes was the fastest (280 pmol/mg/s), and its time course showed a neat overshoot before steady state. The results suggest that either preincubation with Ca2+ plus Mg2+ or calmodulin accelerated phosphorylation shifting toward E1 the equilibrium between the E1 and E2 conformers of the enzyme. K+ had no effect on the initial rate of phosphorylation and lowered by 40% the steady-state level of phosphoenzyme in the absence of Mg2+. Phosphorylation is not rate-limiting for the overall reaction since its initial rate was always higher than ATPase activity. In the absence of K+, the turnover of the phosphoenzyme was 2000 min-1, which is close to the values for other transport ATPases. PMID- 2972721 TI - Phosphoenzymes formed from Mg.ATP and Ca.ATP during pre-steady state kinetics of sarcoplasmic reticulum ATPase. AB - We have investigated here the pre-steady state kinetics of sarcoplasmic reticulum ATPase incubated under conditions where significant amounts of Mg.ATP and Ca.ATP coexist, both of them being substrates for the ATPase. We confirmed that these two substrates are independently hydrolyzed by the ATPase, which thus apparently catalyzes Pi production by two simultaneous and separate pathways. External calcium (or the Ca2+/Mg2+ ratio) determines the extent to which Ca2+ or Mg2+ is bound at the phosphorylation site, while internal calcium controls the rate of processing of both the slow, calcium-containing and the fast, magnesium containing phosphoenzyme. Time-dependent binding of calcium at the catalytic site is correlated with the observed burst of Pi liberation, which therefore results from reequilibration during pre-steady state of magnesium- and calcium-containing phosphoenzyme pools. Independently of direct exchange of metal at the catalytic site, ADP produced by the hydrolysis reaction contributes to reequilibration of these pools through reversal of phosphorylation by the ATP-ADP exchange pathway. PMID- 2972722 TI - A new major transmembrane glycoprotein, gp155, in goat erythrocytes. Isolation and characterization of its association to cytoskeleton through binding with band 3-ankyrin complex. AB - Erythrocyte membranes from goat contain a considerable amount, more than 10% of the total amount, of a glycoprotein with Mr = 155,000 (gp155) on sodium dodecyl sulfate-polyacrylamide electrophoresis gel. This report describes the first isolation and characterization of gp155. This gp155 has major trypsin-sensitive sites at each side of the plasma membrane to generate membrane-bound fragments, indicating that the gp155 spans the lipid bilayer several times. This protein consists of a single polypeptide containing about 1,200 amino acid residues corresponding to Mr = 134,000 and some complex type N-linked oligosaccharide chains. A fraction (15-20%) of the gp155 is recovered in nonionic detergent extracted ghosts along with 25-30% of band 3 and other cytoskeletal proteins and is completely released into solution by extraction with 1 M KCl. Immunoprecipitation with anti-gp155 and anti-ankyrin antibodies of detergent solubilized membranes separated on a gel permeation chromatography column showed that a part of the gp155 is tightly linked to band 3 with a molar ratio of 1:2 to 1:3. This gp155-band 3 complex in turn is associated to ankyrin through the binding of band 3 to ankyrin. These data indicate that, in native erythrocyte membranes, as well as in detergent solution, gp155 could play a physiological role in controlling cellular integrity and elasticity by forming the gp155-band 3 ankyrin complex. Partial amino acid sequences of the tryptic peptides are also determined. PMID- 2972723 TI - The small nuclear ribonucleoprotein E protein gene contains four introns and has upstream similarities to genes for ribosomal proteins. AB - The human small nuclear ribonucleoprotein E protein is an 11,000-dalton basic protein which is an integral component of several small nuclear ribonucleoprotein complexes involved in RNA processing reactions. Sequence analysis of the E protein multigene family reveals that at least one gene for this component of the RNA splicing machinery is interrupted by four introns. The exons of this gene are identical to two cDNA clones isolated from independent tissue sources and span approximately 9 kilobase pairs. Primer extension data indicated the presence of two major transcription start sites. The upstream region of the small nuclear ribonucleoprotein E protein gene does not contain TATA or CCAAT sequences within 175 nucleotides of the transcription start sites. However, the proximal upstream region does contain several similarities to the promoter regions of both snRNA genes and vertebrate ribosomal protein genes. PMID- 2972725 TI - Vertebral fractures without neurological deficit. A long-term follow-up study. AB - Twenty-five patients, who did not have osteoporosis and who were between the ages of seventeen and sixty years, were treated for one or more stable compression fractures of a vertebra with compaction of less than 50 per cent and without an associated neurological deficit. The patients were followed for a minimum of nine years. Associated vertebral fractures (12 per cent) were identified during the first three months after the injury, but no deformity progressed after three months. Radiographic changes of degenerative disc disease were evident in eight patients, but the changes did not correlate with symptoms or with level of activity. With one exception, all patients functioned as well as uninjured subjects of comparable age. Patients who have a compression fracture of a vertebra should have serial radiographs made for at least three months to ensure that all fractured vertebrae are identified and to document any progression of deformity. PMID- 2972724 TI - Clustering of ligand on the surface of a particle enhances adhesion to receptor bearing cells. AB - Human leukocytes express a receptor that mediates the binding of cells and particles coated with C3bi, a fragment of the third component of complement. Previous data indicate that the capacity of this receptor to mediate binding is regulated by changes in its aggregation state. Randomly distributed receptors bind ligand very inefficiently, but stimulation of polymorphonuclear leukocytes with phorbol esters causes a ligand-independent clustering of the receptors in the membrane, and the clustered receptors avidly bind C3bi-coated cells (1). We examined whether the aggregation state of surface-bound ligands also affects the efficiency of binding between receptors and ligands. We found that erythrocytes bearing C3bi in clusters were bound by both macrophages and polymorphonuclear leukocytes far more avidly than erythrocytes bearing the same number of ligands in random array. We made similar observations with erythrocytes coated with C3b, a ligand that is recognized by a separate receptor. Our observations show that the ability of a receptor-bearing cell to bind particles coated with the corresponding ligands is dramatically affected by the distribution of ligand on the surface of the particle. Cell-cell interactions may thus be regulated by alterations in the two-dimensional distribution of receptors and ligands on opposing cell surfaces. PMID- 2972726 TI - Idiopathic dissecting aortic aneurysm associated with pain in the back in an adolescent. Report of a case. PMID- 2972727 TI - Continuous or bolus chemotherapy with 5-fluoro-2'-deoxyuridine in transplanted experimental liver tumors? AB - The effect of continuous and discontinuous locoregional chemotherapy with Floxuridine (FUdR) was studied in a standardized and controlled animal model, using the transplantable Novikoff hepatoma in Sprague-Dawley rats. The liver was perfused after transplantation with a total of 420 mg/kg FUdR, via a fully implanted osmotic minipump or miniport and catheter in the hepatic artery, either continuously (n = 22) from day 5 to day 12, or discontinuously (n = 28) on days 5 and 8. Viable tumor volume and peritumorous cell infiltration were measured. No reduction in tumor volume was attained using discontinuous therapy, in contrast to a highly significant reduction using continuous therapy (P less than 0.001). Significantly less cell infiltration was found after discontinuous than after continuous therapy. In conclusion, continuous locoregional chemotherapy with FUdR was the superior administration method on measurable tumor effect, when compared to discontinuous infusion. PMID- 2972728 TI - Isolated interruption of the aortic arch. AB - We report a case of a 4-year-old girl with interruption of the aortic arch without a patent ductus arteriosus or intracardiac shunts. The anomaly was successfully corrected with the use of a 14 mm Dacron prosthetic graft to bypass the interrupted segment. The postoperative course was uneventful. PMID- 2972729 TI - Sertoli cell processes have axoplasmic features: an ordered microtubule distribution and an abundant high molecular weight microtubule-associated protein (cytoplasmic dynein). AB - Microtubules in the cytoplasm of rat Sertoli cell stage VI-VIII testicular seminiferous epithelium were studied morphometrically by electron microscopy. The Sertoli cell microtubules demonstrated axonal features, being largely parallel in orientation and predominantly spaced one to two microtubule diameters apart, suggesting the presence of microtubule-bound spacer molecules. Testis microtubule associated proteins (MAPs) were isolated by a taxol, salt elution procedure. Testis MAPs promoted microtubule assembly, but to a lesser degree than brain MAPs. High molecular weight MAPs, similar in electrophoretic mobilities to brain MAP-1 and MAP-2, were prominent components of total testis MAPs, though no shared immunoreactivity was detected between testis and brain high molecular weight MAPs using both polyclonal and monoclonal antibodies. Unlike brain high molecular weight MAPs, testis high molecular weight MAPs were not heat stable. Testis MAP composition, studied on postnatal days 5, 10, 15, and 24 and in the adult, changed dramatically during ontogeny. However, the expression of the major testis high molecular weight MAP, called HMW-2, was constitutive and independent of the development of mature germ cells. The Sertoli cell origin of HMW-2 was confirmed by identifying this protein as the major MAP found in an enriched Sertoli cell preparation and in two rat models of testicular injury characterized by germ cell depletion. HMW-2 was selectively released from testis microtubules by ATP and co purified by sucrose density gradient centrifugation with MAP-1C, a neuronal cytoplasmic dynein. The inhibition of the microtubule-activated ATPase activity of HMW-2 by vanadate and erythro-(2-hydroxy-3-nonyl)adenine and its proteolytic breakdown by vanadate-dependent UV photocleavage confirmed the dynein-like nature of HMW-2. As demonstrated by this study, the neuronal and Sertoli cell cytoskeletons share morphological, structural and functional properties. PMID- 2972730 TI - Isolated beta-heavy chain subunit of dynein translocates microtubules in vitro. AB - Our goal was to assess the microtubule translocating ability of individual ATPase subunits of outer arm dynein. Solubilized outer arm dynein from sea urchin sperm (Stronglocentrotus purpuratus) was dissociated into subunits by low ionic strength buffer and fractionated by zonal centrifugation. Fractions were assessed by an in vitro functional assay wherein microtubules move across a glass surface to which isolated dynein fractions had been absorbed. Microtubule gliding activity was coincident with the 12-S beta-heavy chain-intermediate chain 1 ATPase fractions (beta/IC1). Neither the alpha-heavy chain nor the intermediate chains 2 and 3 fractions coincided with microtubule gliding activity. The beta/IC1 ATPase induced very rapid gliding velocities (9.7 +/- 0.88 micron/s, range 7-11.5 micron/s) in 1 mM ATP-containing motility buffers. In direct comparison, isolated intact 21-S outer arm dynein, from which the beta/IC1 fraction was derived, induced slower microtubule gliding rates (21-S dynein, 5.6 +/- 0.7 micron/s; beta/IC1, 8.7 +/- 1.2 micron/s). These results demonstrate that a single subdomain in dynein, the beta/IC1 ATPase, is sufficient for microtubule sliding activity. PMID- 2972732 TI - Functionally distinct laminin receptors mediate cell adhesion and spreading: the requirement for surface galactosyltransferase in cell spreading. AB - The molecular mechanisms underlying cell attachment and subsequent cell spreading on laminin are shown to be distinct form one another. Cell spreading is dependent upon the binding of cell surface galactosyltransferase (GalTase) to laminin oligosaccharides, while initial cell attachment to laminin occurs independent of GalTase activity. Anti-GalTase IgG, as well as the GalTase modifier protein, alpha-lactalbumin, both block GalTase activity and inhibited B16-F10 melanoma cell spreading on laminin, but not initial attachment. On the other hand, the addition of UDP galactose, which increases the catalytic turnover of GalTase, slightly increased cell spreading. None of these reagents had any effect on cell spreading on fibronectin. When GalTase substrates within laminin were either blocked by affinity-purified GalTase or eliminated by prior galactosylation, cell attachment appeared normal, but subsequent cell spreading was totally inhibited. The laminin substrate for GalTase was identified as N-linked oligosaccharides primarily on the A chain, and to a lesser extent on B chains. That N-linked oligosaccharides are necessary for cell spreading was shown by the inability of cells to spread on laminin surfaces pretreated with N-glycanase, even though cell attachment was normal. Cell surface GalTase was distinguished from other reported laminin binding proteins, most notably the 68-kD receptor, since they were differentially eluted from laminin affinity columns. These data show that surface GalTase does not participate during initial cell adhesion to laminin, but mediates subsequent cell spreading by binding to its appropriate N-linked oligosaccharide substrate. These results also emphasize that some of laminin's biological properties can be attributed to its oligosaccharide residues. PMID- 2972731 TI - Immunogold localization of the regulatory subunit of a type II cAMP-dependent protein kinase tightly associated with mammalian sperm flagella. AB - We have shown previously that the regulatory subunit (RII) of a type II cAMP dependent protein kinase is an integral component of the mammalian sperm flagellum (Horowitz, J.A., H. Toeg, and G.A. Orr. 1984. J. Biol. Chem. 259:832 838; Horowitz, J.A., W. Wasco, M. Leiser, and G.A. Orr. 1988. J. Biol. Chem. 263:2098-2104). The subcellular localization of this flagellum-associated RII in bovine caudal epididymal sperm was analyzed at electron microscope resolution with gold-conjugated secondary antibody labeling techniques using anti-RII monoclonal antibodies. By immunoblot analysis, the flagellum-associated RII was shown to interact with mAb 622 which cross reacts with both neural and nonneural isoforms of RII. In contrast, a neural specific monoclonal antibody (mAb 526) failed to interact with flagellar RII. In the midpiece of the demembranated sperm tail, gold label after mAb 622 incubation was primarily associated with the outer mitochondrial membrane. Although almost all specific labeling in the midpiece can be assigned to the mitochondria, in the principal piece, there is some labeling of the fibrous sheath. Labeling of the outer dense fibers and the axoneme was sparse. Specific labeling was virtually absent in the sperm head. Sections of sperm tails incubated in the absence of primary antisera or with mAb 526 showed little labeling. A beta-tubulin monoclonal antibody localized only to the 9 + 2 axoneme. These results raise the possibility that a type II cAMP-dependent protein kinase located at the outer mitochondrial membrane plays a role in the direct cAMP stimulation of mitochondrial respiration during sperm activation. PMID- 2972734 TI - Genome interactions which influence DNA palindrome mediated instability and inviability in Escherichia coli. AB - The interaction of three factors determine the detrimental effect of a palindromic DNA sequence in Escherichia coli cells. The first is the nature of the palindrome (its length, extent of central asymmetry and perhaps its base sequence), the second is the genotype of the host cell and the third is the replicon within which it is located. In this paper we extend the genetic and physical characterization of lambda bacteriophages carrying a palindrome of approximately 560 base pairs. We also show that a palindrome of approximately 110 base pairs which can be cloned in a plasmid cannot be cloned in a phage M13 derivative. These observations are relevant to the choice of vectors used in the cloning of eukaryotic DNA containing palindromic sequences. PMID- 2972733 TI - Macrophage interactions with laminin: PMA selectively induces the adherence and spreading of mouse macrophages on a laminin substratum. AB - The ability of thioglycollate (TG)-elicited mouse peritoneal macrophages to adhere to a laminin substratum has been studied. These cells do not adhere to laminin-coated (20 micrograms/ml) surfaces, but the addition of phorbol myristate acetate (PMA; 50 ng/ml) results in their rapid adherence and spreading on this substratum. TG-elicited and PMA-activated macrophages, however, can bind soluble laminin. Macrophages adhere to fibronectin-coated surfaces and tissue culture plastic without PMA stimulation, and PMA does not increase the number of cells that adhere to these surfaces. The predominant surface proteins that bind specifically to laminin-Sepharose exhibit an Mr of 67 and 36 kD, but the expression of these proteins does not increase after PMA stimulation. Laminin receptor antibodies immunoprecipitate the 67-kD protein from radiolabled surface lysates and are capable of blocking macrophage adherence to a laminin substratum. Indirect immunofluorescence microscopy indicates that PMA stimulation does not increase receptor expression, but that it may induce the aggregation of the receptor on the cell surface. PMA stimulation also promotes macrophage spreading and induces a reorganization of the actin cytoskeleton. Taken together, these data indicate the mechanism by which PMA promotes macrophage adherence to laminin does not involve increased 67-kD receptor surface expression, but that it is related to the changes in cytoskeletal and receptor surface organization that occur in response to PMA stimulation. PMID- 2972735 TI - Facet syndrome and the relief of low back pain. PMID- 2972736 TI - In vitro inhibition of human herpesvirus-6 by phosphonoformate. AB - HSB-2 cell cultures productively infected with human herpesvirus-6 were treated with the antiviral drugs phosphonoformic acid (PFA), acyclovir (ACV), and gancyclovir (DHPG). ACV and DHPG showed significant toxic effects on uninfected HSB-2 cells, yet only incompletely inhibited viral expression upon infection of the cells. PFA, however, showed little direct toxicity on HSB-2 cells while viral replication was inhibited significantly. PMID- 2972737 TI - Phenotypic analyses and concanavalin-A-induced suppressor cell dysfunction of intrathyroidal lymphocytes from patients with Graves' disease. AB - The expression of phenotypic markers and Concanavalin-A-induced suppressor activity was compared among mononuclear cells isolated from thyroid glands and peripheral blood of thionamide-treated patients with hyperthyroid Graves' disease and peripheral blood from normal subjects. Intrathyroidal lymphocytes were obtained by two different methods (TG-1 and TG-2 cells), gradient centrifugation of supernatants of minced thyroid tissue and overnight culture of thyroid debris after mechanical disaggregation and enzymatic digestion, respectively. The percentages of CD3+ cells (all mature T cells) among peripheral blood and TG-1 and TG-2 cells from Graves' patients were similar, but the percentages of B1+ cells (pan B cells) among the TG-1 and TG-2 cells were markedly increased compared to that in peripheral blood. The percentages of CD4+ cells among the TG 1 and TG-2 cells were significantly less than that in peripheral blood. The percentages of CD4+2H4+ cells among CD4+ cells in TG-1 and TG-2 cells also were significantly less than that in peripheral blood. The percentage of CD4+4B4+ cells among CD4+ cells in thyroid glands was markedly higher than that in peripheral blood. The percentages of CD8+ cells and CD8+CD11b- cells (cytotoxic T cells) in thyroid glands were significantly higher than those in peripheral blood from Graves' patients and peripheral blood from normal subjects. The CD8+CD11b+ cells were subdivided into two subpopulations on the basis of CD8 antigen density. The percentage of dull CD8+CD11b+ cells (natural killer cells) among TG 2 cells was lower than that in peripheral blood, but there was no significant difference in bright CD8+CD11b+ cells (suppressor-effector T cells) between thyroid glands and peripheral blood. The percent suppression induced by Concanavalin-A in both TG-1 and TG-2 cells was significantly decreased compared with that in peripheral blood. These results suggest that impairment of suppressor cell activity and an increased number of B cells exist in thyroid glands of patients with Graves' disease compared to those in peripheral blood. It, thus, appears likely that both B cell hyperactivity and suppressor T cell dysfunction may induce excess production of autoantibodies in the thyroid glands of such patients. PMID- 2972738 TI - Effect of excess endogenous androgens on bone density in young women. AB - To determine whether endogenous androgens influence bone density in young women, we studied 27 normal women and 19 women with androgen excess, as defined by increased serum bioavailable testosterone (bio T) concentrations. The women ranged from 21-48 yr of age. The 2 groups were comparable with respect to age, anthropomorphic measures, nutrition, gynecological history, and serum cortisol and estradiol levels. Trabecular (lumbar) and cortical (radial) bone density were quantitated by computerized tomography and single photon absorptiometry, respectively. Serum obtained during the follicular phase of the cycle was assayed for bio T, total T, dehydroepiandrosterone sulfate, androstenedione (Adione), and 3 alpha-androstanediol glucuronide (3-Adiol-G). Trabecular bone density was significantly higher in the androgen excess group [172 +/- 7 (+/- SE) vs. 153 +/- 5 mg/mL; P = 0.03]; controlling for serum Adione (but not for serum bio T, total T, dehydroepiandrosterone sulfate, or 3 alpha-androstanediol glucuronide, or 3 Adiol-G) abolished this difference. Similarly, serum Adione correlated more strongly than the other androgens with trabecular bone density (r = 0.31; P = 0.03). Average cortical bone density was not higher in the androgen excess group (0.740 +/- 0.014 vs. 0.722 +/- 0.008 g/cm2; P = 0.27). Among the 27 normal women, cortical density was correlated to serum bio T (r = 0.47; P = 0.01) and total T (r = 0.53; P = 0.004), but not to the other androgens. We conclude that supraphysiological levels of endogenous androgens are associated with increased trabecular bone density in young women. Serum Adione appeared to be the best marker for the impact of androgen on trabecular density. Among normal women, cortical bone density was related to serum T. PMID- 2972739 TI - Correlation of serum 3 alpha-androstanediol glucuronide with acne and chest hair density in men. AB - Serum 3 alpha-androstanediol glucuronide (3 alpha-Adiol-G) is considered to be an indicator of peripheral tissue androgen metabolism. Precursor circulating androgens are converted in peripheral tissue to dihydrotestosterone (DHT), which is ultimately metabolized to 3 alpha-Adiol-G and secreted from the cell. Elevated serum 3 alpha-Adiol-G concentrations have been reported in women in hyperandrogenic states. We studied 44 consecutive male medical students for chest hair density, acne, and serum dehydroepiandrosterone sulfate (DHEA-S), total testosterone (total T), free and albumin-bound (bioavailable) T (bio T), and 3 alpha-Adiol-G concentrations. Although there was considerable overlap of serum 3 alpha-Adiol-G values among the groups defined by hair density or acne scores, we found statistically significant correlations between serum 3 alpha-Adiol-G and chest hairiness (P = 0.0034), acne (P = 0.0005), and a combined chest hairiness and acne score (P = 0.0018). There was no significant correlation between these clinical parameters and the levels of precursor androgens. There was, however, a strong correlation between serum 3 alpha-Adiol-G and bio T (P = 0.0005), suggesting that in men serum 3 alpha-Adiol-G levels may be dependent upon available free and albumin-bound T. The correlations in men of serum 3 alpha Adiol-G with chest hair density, acne, and the hairiness and acne index supports the hypothesis that the serum levels of 3 alpha-Adiol-G reflect the extent of androgen action in peripheral tissues. PMID- 2972740 TI - Ectopic expression of HLA class II antigens on thyroid follicular cells: induction and transfer in vitro by autologous mononuclear leukocytes. AB - Ectopic expression of HLA class II antigens by thyroid follicular cells (TFC) might trigger the immune recognition of TFC-specific surface constituents by self reactive T helper/inducer lymphocytes, with the consequent generation of organ specific autoimmunity. To explore the alternative possibilities underlying this ectopic expression, i.e. that it either reflects a primary abnormality of the TFC or is secondarily induced by the autoimmune attack itself, we employed immunofluorescence staining of monolayers cultured from thyroid tissue of patients with autoimmune thyroid disease as well as neoplastic, nodular, and normal thyroid tissues to assess the role that autologous mononuclear leukocytes (MNC) and serum factors play in HLA class II induction and modulation in vitro. In all Graves' disease (GD) tissues that initially displayed HLA-DR (DR) antigens, this expression was transient. Primary TFC cultures from tissues with tumor-associated chronic thyroiditis (CT) showed more widespread and persistent DR expression than did cultures from GD tissues. However, in contrast with the findings in GD, there was no correlation between ectopic DR expression on TFC from the CT areas and the presence of organ-specific autoimmune markers. DR expression by TFC was detected in only one of the five nontoxic nodules studied, and involved a small proportion (approximately 20%) of the TFC. A similarly low proportion of DR-positive TFC was found in cultures from two papillary carcinomas, while much stronger DR expression was seen on TFC cultured from the contralateral lobes affected with CT. Only one of four follicular adenomas expressed DR on about 35% of the cultured TFC. In contrast with the rapid extinction of DR expression on TFC cultured from GD tissues after depletion of infiltrated MNC, DR antigen loss did not occur or was significantly delayed when parallel TFC monolayers were cocultured in the presence of autologous MNC from the intrathyroidal infiltrates. Coculture of DR-negative TFC from normal tissues with autologous peripheral blood MNC in individuals without detectable thyroid autosensitization also resulted in pronounced induction of DR expression on the TFC. Purified T lymphocytes from the same peripheral blood MNC preparations, however, did not induce DR expression on these normal TFC. Similarly, MNC obtained from two follicular adenomas, mostly macrophages, did not induce DR expression when cultured with autologous TFC from the same lesions. Addition of 15% autologous serum did not prevent progressive extinction of DR expression on TFC cultured from GD or tumor-associated CT tissues after depletion of infiltrated MNC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2972741 TI - Production of and response to interleukin-2 in Graves' disease. AB - Interleukin-2 is a lymphokine which is believed to play a central role in the regulation of the immune response. The production of and response to interleukin 2 were determined in hyperthyroid Graves' patients together with thyroid function and serum thyrotropin receptor antibody, a marker of autoimmune activity. Interleukin-2 production by mitogen-induced peripheral blood mononuclears was markedly low in 24 of 29 patients when compared to controls. Five patients in remission had normal values. In nine patients followed during antithyroid drug therapy, interleukin-2 production returned gradually to normal levels within 4-6 months. This rise and the concomitant decrease in serum thyrotropin receptor antibody correlated with the decline in the free thyroxin index. Antithyroid drugs and triiodothyronine had no effect on interleukin-2 production in vitro. Mitogen-activated mononuclears from hyperthyroid Graves' patients did not proliferate as well as the controls in response to interleukin-2. However, seven patients treated with antithyroid drugs and three in remission responded normally. Flow cytometry using anti-Tac antibody revealed that the interleukin-2 receptor density on mononuclears from five patients was low. This parameter was normal in treated patients and those in remission. We conclude that the production of and response to interleukin-2 by peripheral blood mononuclears from hyperthyroid Graves' patients are poor, the latter being due to impaired receptor expression. Both aberrations are restored to normal by antithyroid drug therapy or in remission. The relative roles of the autoimmune process and thyroid function in modulating the interleukin-2 pathway and the question of whether antithyroid drugs act directly or through thyroid inhibition remain to be clarified. PMID- 2972743 TI - Disappearing optociliary shunt vessels and neonatal hydrocephalus. AB - This is a report of two cases of optociliary shunt vessels in patients with neonatal hydrocephalus. Complete disappearance of the optociliary shunt vessels occurred after surgical procedures to normalize intracranial pressure. The clinicopathological significance, prognosis, and treatment of patients with acquired optociliary shunt vessels is discussed. This is also the first report of the association of optociliary shunt vessels and neonatal hydrocephalus. PMID- 2972742 TI - Dendritic cell-lymphocyte clusters that form spontaneously in rheumatoid arthritis synovial effusions differ from clusters formed in human mixed leukocyte reactions. AB - Lymphocytes cluster about dendritic cells (DC) spontaneously in 48 h cultures of rheumatoid arthritis synovial fluid (RA SF) mononuclear cells and in peripheral blood autologous or allogeneic mixed leukocyte reactions. In the latter case, the clusters are predominantly CD4+ T cells (T4/T8 greater than 5) and with time progress in blastic cells that express IL-2 (Tac) and/or transferrin (T9) receptors. In contrast, the clusters in RA SF cultures have a T4/T8 ratio of less than 1 and a majority of the T8 cells coexpress the Leu 7 marker. T cells in these clusters remain inert and with time the clusters disintegrate. Addition of IL-1, IL-2, or IFN-gamma alone or in combination had no effect on RA SF clusters but T cells became blastic when exposed to 10% RA SF. Mixing experiments using RA SF DC with normal T cells and RA T cells with normal DC show that both RA SF DC and T cells are immunofunctional. In addition, clusters of RA SF from a patient with active tuberculosis proliferated vigorously to PPD. Therefore, the unique RA SF cluster profile may reflect the memory nature of the RA SF T cells resulting in a paucity of T cells that are responsive to autologous stimulation. However, an immunosuppressive role for the double-labeled (CD8 and Leu 7) cells has not been excluded. PMID- 2972745 TI - Loss of accommodation produced by peristriate lesion in man? AB - We observed a bilateral accommodative paresis associated with a spontaneous parieto-occipital hematoma in a 37-year old patient. There was no clinical or computerized tomography (CT) evidence of transtentorial herniation or upper brainstem pathology. With resolution of the hematoma, accommodation returned to normal. PMID- 2972744 TI - Abducens nerve palsy and Horner's syndrome revisited. AB - Sympathetic fibers destined for the eye join the abducens nerve for a short distance within the cavernous sinus; thus, a unilateral sixth nerve palsy with an ipsilateral Horner's syndrome is of significant localizing value. We report two cases of cavernous sinus lesions producing an ipsilateral abducens palsy and Horner's syndrome: one case due to an intrinsic and the other to an extrinsic lesion of the sinus region. PMID- 2972746 TI - Bilateral optic neuropathy associated with amiodarone therapy. AB - Amiodarone, an antiarrhythmic agent, has been associated with mild visual loss secondary to papillopathy and papilledema. We report a patient who developed bilateral optic neuropathy 4 weeks after initiation of amiodarone therapy. Nine months later, his vision was 20/50 in the right eye and 20/200 in the left. This report provides additional evidence that amiodarone may cause toxic optic neuropathy. PMID- 2972747 TI - Amiodarone optic neuropathy. PMID- 2972748 TI - Lid-opening apraxia in Wilson's disease. AB - A student was diagnosed as having Wilson's disease only after the severe, intermittent inability to open his eyes led him to seek neurologic evaluation. Although lid-opening apraxia is usually a symptom of diffuse extrapyramidal disease, it has not previously been reported in Wilson's disease. PMID- 2972749 TI - Subjective oscillopsia ("jiggling" vision) presumably due to aminoglycoside ototoxicity. A report of two cases. AB - Following aminoglycoside antibiotic therapy, two patients developed self-limited subjective oscillopsia in the absence of a detectable ocular motility disturbance (nystagmus or opsoclonus). Oscillopsia represents a rare, but highly distressing symptom resulting from disruption of the vestibulo-ocular reflex, producing profound illusory movement of the visual environment. Although the differential diagnosis includes vascular, inflammatory, and structural disorders impacting on either the central or peripheral projections of this brainstem reflex, iatrogenic aminoglycoside ototoxicity was the likely explanation in the two patients presented. Ways of minimizing the risk of aminoglycoside toxicity are briefly reviewed. PMID- 2972751 TI - Pseudo-pseudo-Foster Kennedy syndrome. AB - An 80-year-old woman presented with a classic story and findings of an anterior ischemic optic neuropathy in her left eye. Her right eye had slow and progressive decreased vision, ostensibly secondary to a cataract. However, the right eye showed slight temporal pallor of the optic disc and a superior temporal field defect was found. Her radiologic exam showed a tuberculum sella meningioma extending into the right optic canal compressing the right optic nerve. Two diseases, ischemic optic neuropathy and meningioma, in one patient may be termed a pseudo-pseudo-Foster Kennedy syndrome. PMID- 2972750 TI - Optic neuropathy associated with chronic lymphomatous meningitis. AB - A patient with paranasal sinus lymphoma (recognized retrospectively) developed unilateral, acute, self-limited optic neuritis during the course of chronic lymphocytic meningitis with elevated intracranial pressure and headache. Meningeal symptoms were adequately controlled with analgesics alone for 14 months and corticosteroids alone for a subsequent 11 months without evidence of development of other involvement of organs outside the central nervous system (CNS). Eventually, the visual alteration from optic neuritis prompted a repeat evaluation, which disclosed lymphoma in bone marrow. The subject of paraneoplastic optic neuritis is reviewed. PMID- 2972752 TI - Insight into the recent European literature. A review of studies concerning the visual pathways. PMID- 2972753 TI - The "phantom" optic nerve. Demonstration in CT and MR scans 19 years after resection of optic glioma. PMID- 2972755 TI - Evidence of orbital deformation in indirect optic nerve injury. Weight lifter's optic neuropathy. AB - A 16-year-old boy developed monocular visual loss due to optic neuropathy following frontal head trauma. His trauma was unique in that it consisted of a static loading force to the brow, rather than the dynamic impact more commonly associated with blunt orbital injuries. This case demonstrates the role of isolated deformation of orbital bones in the pathogenesis of indirect optic nerve injury. PMID- 2972754 TI - Congenital glaucoma and buphthalmos in a child with neurofibromatosis. PMID- 2972756 TI - Neuroendocrine effects of diazepam in normal subjects following brief painful stress. AB - The neuroendocrine effects of 5 and 10 mg of oral diazepam were assessed in 10 normal subjects under baseline (prestress) and laboratory-controlled stressful conditions. Although the 10-mg diazepam dose had no effect on cortisol at baseline, it significantly reduced the increase in cortisol produced by 15 minutes of exposure to a painful electrical stimulus. There were no significant effects on growth hormone, beta-endorphin or ACTH, either at baseline or after painful stimulation. PMID- 2972757 TI - An analysis of the speech of five Down's syndrome adolescents. AB - his paper presents an analysis of the speech of five Dutch-speaking adolescent girls with Down's syndrome. The analysis includes a phonetic analysis, a substitution analysis, and a phonological process analysis. The results indicate that the speech errors of Down's syndrome adolescents are for the greater part identical to the error patterns observed in young normal children, supporting the view that the misarticulations in mongoloid subjects are mainly the result of a delay in speech development. PMID- 2972758 TI - Staphylococcus aureus colonization of burrows in erythrodermic Norwegian scabies. A case study of iatrogenic contagion. AB - Scanning electron microscopy demonstrated extensive bacterial colonization of scabies burrows honeycombing the stratum corneum of an elderly woman with erythroderma. Cultures of scybala revealed hemolytic Staphylococcus aureus, possibly responsible for the erythroderma. Epidemiologic data revealed a trail of scabies through two nursing homes and one hospital during the 2-year period that physicians believed she had a drug-induced erythroderma. PMID- 2972759 TI - Cutaneous reactions to vitamin K. AB - In four patients the development of erythematous plaques after vitamin K injections was apparently due to delayed hypersensitivity reactions. The 36 cases described in the literature indicate that most patients with the reaction received vitamin K injections for liver disease and that a late sclerodermatous reaction develops rarely. PMID- 2972760 TI - On the classification of the sign of Leser-Trelat. PMID- 2972761 TI - Erythema multiforme following diphtheria and tetanus toxoid vaccination. PMID- 2972762 TI - Multiple eruptive syringomas associated with Down's syndrome. PMID- 2972763 TI - Polymerization contraction and reaction kinetics of three chemically activated restorative resins. PMID- 2972764 TI - Whole mount technique: an improved hamster ear model to evaluate pharmacologic effects on sebaceous glands. PMID- 2972765 TI - Assessment of the effectiveness of glove use as a barrier technique in the dental operatory. AB - To assess the effectiveness of routine glove use as a barrier technique in the dental operatory, the relationship between frequency of glove use and hepatitis B infection was examined. Hepatitis B screening results and information on infection control practices of 1,109 dentists who attended the 1985 annual session were compiled and statistically analyzed. The results showed that 18% of the dentists wore gloves routinely, 65% wore gloves intermittently, and 17% never wore gloves. Frequency of glove use was related to hepatitis B infection (P less than .01), with dentists who never wore gloves being infected more than twice as often as dentists who wore gloves routinely. Dentists with less frequent glove use and more years in practice had a higher incidence of hepatitis B infection (P less than .0001). PMID- 2972766 TI - Pretreatment management of the patient receiving anticoagulant drugs. AB - This literature review suggests that certain low-risk dental treatment procedures can be performed in patients without altering their anticoagulant medications. Intermediate-risk dental procedures in these patients may be accomplished outside the hospital if the patient's prothrombin time value is within a specified range and if certain techniques are followed. In some cases, temporarily altering the dose of anticoagulant may be necessary. Patients receiving anticoagulant medications should continue to receive high-risk dental treatment in hospitals. PMID- 2972767 TI - The effects of carbon monoxide on persistent changes in young rat heart: cardiomegaly, tachycardia and altered DNA content. AB - Newborn rat pups that inhale 500 ppm carbon monoxide (CO) for 32 days and develop increased heart mass (i.e. cardiomegaly) show persistent cardiomegaly and elevated resting heart rate as adults. Studies were carried out to explore the relationship of these phenomena to CO concentration and initial cardiomegaly using exposures of 350, 500 and 700 ppm CO. Initial cardiomegaly was greater in the right ventricle free wall (RV) than in the left ventricle plus interventricular septum (LV + S) at all three CO concentrations, and RV mass excess increased with CO concentration. Initial RV cardiomegaly was greater at 350 and 700 ppm CO in females than in males. Persistent cardiomegaly in the RV also increased with CO concentration, and was significantly greater in the females. Persistent cardiomegaly in the RV increased with initial cardiomegaly but at a decreasing rate at the higher CO concentrations, and when expressed as a percentage of initial cardiomegaly (i.e. 'efficiency'), the relative response was greatest at 500 ppm CO. For LV + S, efficiency of development of persistent cardiomegaly was greatest at 350 ppm CO. Persistent tachycardia increased with CO concentration in males but failed to do so in females, and was only weakly correlated with degree of persistent cardiomegaly. Thus, persistent cardiomegaly and persistent tachycardia (in males) are related to CO concentration. Myocardial DNA content of 32-day-old juveniles was significantly increased at 350 and 700 ppm CO. Adult DNA content of the RV was significantly elevated at 350 ppm CO (females) and continued to rise with CO concentration. DNA concentrations of the RV and LV were increased at 700 ppm CO.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972768 TI - The link between cytosolic Ca2+ and the Na+-H+ antiport: a unifying factor for essential hypertension. PMID- 2972770 TI - Resistance of primary CD8+ cytotoxic T lymphocytes to lysis by cytotoxic granules from cloned T cell lines. AB - Recent evidence has shown that cloned, murine CTL cell lines are resistant to the cytotoxic components of the toxic granules they release upon specific interaction with their target cells. Inasmuch as the resistance might be due to selection in culture over many months by repeated exposure to these cytolytic components (which are released repeatedly as a result of the cultured CTL being periodically stimulated by target cells), we asked whether primary CTL are also resistant. The primary CTL were elicited in vivo by i.p. injection of allogeneic tumor cells or in vitro by 5- to 6-day MLC or by 48-h exposure to the lectin Con A. The responding cells were separated into purified CD8+ (i.e., CD4-, CD8+) and purified CD4+ (i.e., CD4+, CD8-) T cell populations that were analyzed for cytolytic activity and for resistance to lysis by toxic secretory granules derived from cloned CTL cell lines. The CD8+ T cells were highly cytolytic and relatively resistant; they retained their cytolytic activity and were lysed to a minimal extent (0 to 10%) by quantities of isolated granules that lysed 80 to 90% of the P815 tumor cell line (tested as a representative standard cell line). The CD4+ T cells, in contrast, had only minimal cytolytic activity and were far more susceptible to granule-mediated lysis. Although the resistance of primary CD8+ T cells is impressive, it is not as pronounced as the resistance of the cloned CTL cell lines, indicating that during long-term culture there is some selection for increased resistance to granule-mediated lysis. In contrast to T cells (especially CD8+ T cells), Ia+ macrophages, isolated from primary immune peritoneal exudates, were highly susceptible to granule-mediated lysis. PMID- 2972769 TI - Differential activation requirements for virgin and memory T cells. AB - Most studies of the activation requirements for T cells have used either T cell lines or populations of normal T cells that consist of a mixture of virgin and Ag primed T cells. These two subpopulations of T cells can now be distinguished in humans by their reactivity with mAb. The anti-CD45R antibody HB10 identifies virgin T cells (T degrees) that are non-reactive to recall Ag and relatively poor at providing help for B cell differentiation. Conversely, memory T cells (T') that can react to recall Ag and enhance Ig production are non-reactive with anti CD45R, but can be identified with the UCHL1 antibody. We have used these antibodies to separate the T degrees and T' populations and examine their activation requirements. On activation CD45R+ cells rapidly began to lose the CD45R Ag and express the UCHL1 Ag in increased amounts, whereas the UCHL1+ cells retained this phenotype. Both populations responded to PHA in the presence of monocytes, but when triggered by an antibody to CD3 only the T' cells were induced to express IL-2R, produce IL-2, and to proliferate. The T degrees population of cells remained relatively quiescent by all of these parameters. However, anti-CD3 stimulation conditioned the T degrees cells for IL-2 responsiveness, inasmuch as the addition of rIL-2 resulted in significant IL-2R expression and proliferation. When the CD4+ T degrees and CD4+ T' subpopulations were isolated and examined in the same assays similar results were obtained. The data indicate that fundamental differences exist in the triggering requirements for T degrees and T' cells. PMID- 2972771 TI - Bone marrow cells from allogeneic bone marrow chimeras inhibit the generation of cytotoxic lymphocyte responses against both donor and recipient cells. AB - When added to a mixed lymphocyte culture, bone marrow cells suppress the generation of CTL activity against H-2 Ag shared by the BM cells and the stimulator cells. These cells have been referred to as veto cells and are thought to play a role in maintaining self-tolerance. We analyzed the H-2 specificity of the suppression expressed by the veto cells from H-2 incompatible bone marrow chimeras, because lymphocytes of such chimeras had been shown to be tolerant to both donor and recipient Ag when tested by CTL responses. We found that the bone marrow cells of such chimeras which were featured by non-T and non-B cell characteristics inhibited the generation of CTL directed against either donor or recipient Ag, but not against third-party Ag. These observations suggest that in allogeneic chimeras the veto or veto-like cells alter the inhibitory specificity exhibited in the recipient microenvironment and indicate that these cells are directly involved in the induction and maintenance of self-tolerance. PMID- 2972772 TI - Regulation of antibody production by helper T cell clones in experimental autoimmune myasthenia gravis. AB - Ten acetylcholine receptor (AChR)-specific T cell clones from Lewis rats were studied. These clones had various AChR subunit and peptide specificities, and proliferated in response to antigen on appropriate APC. All the T cell clones were CD4+CD8- and OX22-, helped anti-AChR antibody production by AChR-primed lymph node B cells, and could secrete IL-2. However, several lines of evidence suggested that IL-2 was not the lymphokine that mediated T cell help. B cells primed with native AChR and then exposed in culture to very low concentrations of native AChR effectively presented the Ag to the T cell lines, presumably due to uptake via Ag receptors, but primed B cells were no more effective than were non specific APC at presenting a synthetic AChR peptide which is recognized by AChR specific T cells but not by AChR-specific B cells. Increasing AChR doses produced an antibody production response that was bell shaped and low doses stimulated, whereas higher AChR concentrations suppressed the antibody production response. Evidence suggested that AChR exerted its inhibitory effect through the T cells, but not via IL-2. PMID- 2972773 TI - IL-4 induces loss of B lymphocyte Fc gamma R II ligand binding capacity. AB - Murine B lymphocytes cultured for 24 h with rIL-4 lost (mean reduction of 88%, range 81 to 96%) the capacity to bind Ag-IgG antibody complexes to B lymphocytes as assessed by flow microfluorometry. This effect was specific in that it was not seen with IL-1, IL-2, or IFN-gamma; IL-4 did not have a similar effect on other B lymphocyte membrane molecules; and the effect was completely prevented by anti-IL 4 (mAb 11B11). More than 60% inhibition of the binding of complexes was seen with as little as 1 U/ml of IL-4 although maximal inhibition was seen with greater than or equal to 30 U/ml. IL-4-induced inhibition of the binding of complexes was time dependent (the effect was first seen after 8 h and was not maximal until 24 h), temperature dependent (it did not occur at 4 degrees C), and reversible (B lymphocytes that had lost the ability to bind complexes due to IL-4 regained this capacity when re-cultured for 24 h in the absence of IL-4). The effect could be partially prevented by IFN-gamma. The inability to bind complexes appeared to be mainly due to an alteration of Fc gamma R (Fc Receptors) II rather than down regulation of receptor expression because IL-4 induced only a moderate reduction in the binding of two Fc gamma R II specific mAb (20% for 2.4G2 and 32% for K9.361). The IL-4-induced loss of binding of complexes to B lymphocyte Fc gamma R II appears to be a novel form of receptor regulation (function rather than expression), and likely plays a role in the up-regulation of B lymphocytes by IL 4 by preventing Fc gamma R II-mediated inhibition of B lymphocyte responses. PMID- 2972774 TI - Characterization of T helper 1 and 2 cell subsets in normal mice. Helper T cells responsible for IL-4 and IL-5 production are present as precursors that require priming before they develop into lymphokine-secreting cells. AB - We have shown that the requirements for the production of IL-4 and IL-5 by normal L3T4+T cells from murine spleen are very different from those for the production of IL-2. Secretion of detectable quantities of IL-4 and IL-5 and induction of the mRNA for each lymphokine occurs in vitro only after cells are primed and re stimulated. This priming can be achieved by mitogens (Con A), by antibodies to the TCR (anti-T3) or by stimulation with alloantigen. In contrast, requirements for induction of lymphokine production after priming resemble those for initial production of IL-2. Thus the majority of T cells of helper phenotype that have the potential to become IL-4- and IL-5-secreting T cells, exist in the form of precursors requiring stimulation and several days of culture as well as re stimulation with mitogen or Ag before they become detectable as lymphokine secreting cells. In contrast, among fresh CD4+T cells, secretion of IL-2, IL-3, granulocyte/macrophage CSF, and IFN-gamma is easily detected within 24 h of stimulation with mitogen or Ag. These observations establish that distinct phenotypes of Th cells are found at different times after stimulation and support the concept that synthesis and secretion of different lymphokines or groups of lymphokines are regulated independently. Furthermore the patterns of lymphokines secreted by fresh vs primed Th cells, which largely correspond to the patterns that have been used to define the Th1 and Th2 subsets among Th cell lines, provides evidence that different subsets of normal T cells exist that may correspond to these designations. Secretion of different lymphokines by two subsets of Th cells at different times in an immune response, and perhaps in different places, suggests a model in which the ratio of the two T cell subsets (Th1 vs Th2) and state of differentiation of each (precursor vs effector), influence or determine the direction of the response, with variations in these parameters leading to differing responses. PMID- 2972775 TI - Phagocytosis of serum-opsonized zymosan down-regulates the expression of CR3 and FcRI in the membrane of human monocytes. AB - The effect of iC3b receptor (CR3)-mediated phagocytosis on the expression of CR (C3b receptor, CR3) and IgG FcR (FcRI, FcRII) has been investigated by using serum-opsonized zymosan as a multivalent ligand for CR3. Sixteen hours after a short (1-h) pretreatment of human monocyte monolayers with zymosan opsonized with human AB serum (250 micrograms/ml), CR3 expression (as assessed by flow cytometric analysis with mAb Mo1) was significantly reduced by 59 +/- 3% (mean +/ SEM, n = 15, p less than 0.001). Concomitant with CR3 down modulation, FcR binding activity (as assessed by binding of IgG-coated E) was also found to be decreased to 41 +/- 4% of control (n = 7, p less than 0.001). Reduced FcR function was paralleled by a decrease in the expression of FcRI (as assessed with mAb 32.2). This FcRI modulation was not caused by zymosan-bound IgG because zymosan opsonized with agammaglobulinemic serum equally down regulated CR3 and FcRI expression. Pretreatment with zymosan opsonized with human AB serum, however, did not change the expression of other IgG and C-binding sites such as FcRII (examined with mAb IV.3 and 2E1) and CR1 (assessed with mAb 57F) as well as of unrelated cell membrane structures (beta 2m, MHC class II). In contrast, co modulation for FcR function and CR3 expression induced by polymeric IgG is accompanied by a decreased expression of FcRII. These data indicate that interaction of a specific receptor with its ligand not only changes the expression of the receptor triggered, but has also a modulating effect on other receptor systems on the same cell. PMID- 2972776 TI - Athymic nude CD4+8- T cells produce IL-2 but fail to proliferate in response to mitogenic stimuli. AB - A comparative study of immune functions of CD4+8- T cells isolated from normal and athymic nude mice by electronic cell sorting was performed. Athymic nude CD4+8- T cells expressed the TCR-associated CD3 molecule but the level of expression was significantly lower than that of normal CD4+8- T cells. Proliferative responses were studied upon stimulation by 1) the T cell mitogen Con A; 2) anti-CD3 mediated cross-linking of the CD3:TCR complex, and 3) the combined action of PMA + ionomycin. All three mitogenic stimuli caused readily detectable cell division in normal (euthymic) CD4+8- T cells. In marked contrast, none of the mitogenic stimuli induced significant proliferation in athymic nude CD4+8- T cells. The failure of athymic nude CD4+8- T cells to proliferate occurred over a wide range of mitogen concentrations and over a 4-day observation period. Neither exogenously supplied rIL-2 or mixed lymphocyte culture supernatant had any effect on the impaired proliferative response by athymic nude CD4+8- T cells. Although IL-2 was produced by athymic nude CD4+8- T cells at a reduced level when compared to normal CD4+8- T cells, it was nevertheless readily detected upon stimulation with either Con A or anti-CD3. Furthermore, stimulation of athymic nude CD4+8- T cells by anti-CD3 induced the expression of the p55 chain of IL-2R on the cell surface. Therefore, despite production of IL-2 and induced expression of IL-2R, athymic nude CD4+8- T cells failed to undergo cell division. PMID- 2972777 TI - Induction of anti-CD8 resistant cytotoxic T lymphocytes by anti-CD8 antibodies. Functional evidence for T cell signaling induced by multi-valent cross-linking of CD8 on precursor cells. AB - The effector function of most MHC class I allospecific CTL is inhibited by anti CD8 mAb. In the present study, we report the surprising observation that multi valent cross-linking of CD8 molecules on precursor cells by specific antibody actively induces the generation of CD8+ class I allospecific CTL whose lytic function is resistant to anti-CD8 antibody inhibition, and actively induces down modulation of cell surface CD8 expression on these cells. In marked contrast, bi valent cross-linking of CD8 inhibits the generation of CD8+ CTL from precursor cells and fails to induce down-modulation of cell surface CD8 expression. These results demonstrate that CD8 can transduce net positive signals, but only when the molecule is extensively cross-linked. PMID- 2972778 TI - IL-1 is an autocrine growth factor for T cell clones. AB - Activation of Th lymphocytes requires that Ag be presented on the surface of accessory cells displaying Ia Ag. A number of studies have concluded that the T cell also requires IL-1 from accessory cells. However, we recently reported that one murine T cell clone (D10.G4.1) produced its own IL-1-like activity after encountering APC (9). In this report, we demonstrate that 1) IL-1 production is a common property of murine T cell clones, 2) T cell IL-1 activity is blocked by anti-IL-1-alpha antiserum, 3) IL-1-alpha mRNA can be directly visualized in individual cloned T cells using in situ hybridization techniques, and 4) IL-1 appears to serve an autocrine role in the activation of T cell clones inasmuch as anti-IL-1-alpha antiserum blocks cell proliferation when the T cell is the only IL-1 source. PMID- 2972779 TI - Direct evidence for the clustered nature of complement receptors type 1 on the erythrocyte membrane. AB - C receptors 1 (CR1) of human E are involved in the transport of C3b-coated immune complexes (IC) in the circulation. Many studies have suggested that the binding of IC to E is multivalent. This would require CR1 to be clustered on the cell membrane, but no direct evidence for such clustering is available. We studied the distribution of CR1 on human E by immunofluorescence and shadow-casting immuno electron microscopy techniques with the use of a monoclonal anti-CR1 antibody followed by FITC- or gold-conjugated second antibodies, respectively. By immunofluorescence, CR1 appeared as small dots (clusters) on fixed and unfixed E prepared either at 4 degrees C or at 37 degrees C. In the same donor, the number of clusters varied extensively from cell to cell (e.g., 1 to 43 clusters/E for a donor with 520 CR1/cell), but the mean number of clusters per cell correlated significantly with the mean number of CR1/cell. These images contrasted with those obtained for Rhesus D (RhD) Ag used as controls (RhD Ag are known to be evenly distributed): only a faint uniform fluorescence was seen despite the presence of 10,000 antigenic sites. As determined by immunocytochemical method, more than 65% of the total gold particles were organized in clusters (2 to 15 gold particles/cluster) whether cells were prefixed or not. Quantitative determinations suggested that each gold particle corresponded to one CR1. The fraction of gold particles grouped into clusters of three or more receptors, the mean size of the clusters, and the maximal size of clusters correlated with the mean number of CR1 per cell. By contrast, RhD Ag were distributed homogeneously (less than 2% gold particles in clusters). These data are the first to demonstrate the preclustered nature of CR1 on E. Such distribution could explain the high binding efficiency of C3b-coated IC to E despite the low number of CR1 per cell. PMID- 2972781 TI - Pleiotropic effects of the Bcg gene. II. Genetic restriction of responses to mitogens and allogeneic targets. AB - The response of Bcgr and Bcgs spleen cells to allogeneic Ag, mitogens, and in a system of oxidative mitogenesis using neuraminidase and galactose oxidase was investigated in two Bcg congenic systems. The Bcgr macrophages supported the MLR across H-2 barrier much better than the Bcgs macrophages. At sub-optimal or optimal doses of mitogens Bcgr mice were higher responders than their Bcgs counterparts. The superior response of Bcgr spleen cells to Con A was further investigated with the aim of identifying the population expressing this phenotype. T cells of either Bcgr or Bcgs type showed equal ability to respond to Con A in the presence of macrophages. Purified splenic macrophages from Bcgr mice contained a significantly greater percentage of Ia+-bearing macrophages compared to Bcgs mice. Splenic macrophages of the Bcgr type were more efficient than their Bcgs counterparts at restoring the Con A response of accessory cell-depleted spleen cells. Resident peritoneal macrophages as well as splenic dendritic cells from Bcgr and Bcgs mice were equally efficient at restoring this response. Glutaraldehyde-fixed Bcgr splenic macrophages were shown to be more efficient than the Bcgs cells at replenishing the response of Con A-unresponsive spleen cells when supplemented with IL-1. PMID- 2972780 TI - Characterization of CD45 and CD45R monoclonal antibodies using transfected mouse cell lines that express individual human leukocyte common antigens. AB - Five cell lines that express one of five isoforms of the human leukocyte common Ag (CD45) were established by transfecting a murine pre-B cell line with leukocyte common Ag cDNA constructs. Using these cell lines, the specificities of CD45 and CD45R mAb were examined. Of the 43 mAb tested, 25 antibodies recognized sequences common to all five isoforms, 16 antibodies recognized isoforms that include exon A encoded sequences, one antibody recognized isoforms that include exon B encoded sequences, and one antibody recognized only the isoform that does not include either exon A, B, or C encoded sequences. PMID- 2972782 TI - Determination of cytotoxic T lymphocyte activity by time-resolved fluorometry using europium-labelled concanavalin A-stimulated cells as targets. AB - Time-resolved fluorometry was used to detect the cytolysis of concanavalin A stimulated blast cells by employing cells stimulated in a mixed lymphocyte culture as effectors. The target cells were labelled with europium diethylenetriaminopentaacetate (EuDTPA) chelates. The results obtained showed that this method, which has been successfully applied to the measurement of natural killer cell activity, was also applicable to the more specific cytotoxic T lymphocyte reaction. The specific release was higher with EuDTPA-labelled target cells than with 51chromium-labelled target cells. As with 51chromium, EuDTPA can be used to distinguish between the cytolysis of specific and non specific target cells. PMID- 2972783 TI - A simple and sensitive bioassay for the detection of IL-2 activity. AB - A simple, one-step quantitative assay for the detection of biologically active interleukin-2 (IL-2) is described. It is based on culture of pure CD3+ T cells which have been positively selected from blood mononuclear cells by particle (M450)-bound anti-CD3 monoclonal antibody (mAb). During culture, activation of the T cells via CD3 will occur, leading to expression of IL-2 receptors but not IL-2 production. By adding IL-2 a proliferative response is evoked, giving a linear dose-response curve for IL-2 concentrations between 0.01-20 U/ml. The cells were unresponsive to IL-1, IL-4, tumor necrosis factor alpha (TNF-alpha), interferon-alpha (IFN-alpha), IFN-gamma, phytohemagglutinin and concanavalin A. The responsiveness to IL-2 was enhanced by TNF-alpha and inhibited by IFN-alpha, while the other tested lymphokines and mitogens did not influence the proliferative response. Antibodies to IL-2 and to IL-2 receptor suppressed the IL 2 response in a dose-dependent manner. The method is both simple and specific, and obviates the necessity for keeping assay cells in long term culture. PMID- 2972785 TI - Endogenous opioids and the exercise-induced augmentation of natural killer cell activity. AB - Based on prior observations that both beta-endorphin and exercise stimulate natural killer (NK) cell activity, we have examined the hypothesis that the release of endogenous opioids during the stress of acute exercise may mediate this NK cell augmentation. Eight healthy young women underwent a maximal bicycle ergometer exercise test with prior in vivo administration of a placebo and an opioid antagonist, naloxone (100 micrograms/kg), in a randomized, blind protocol. Exercise after the placebo injection was accompanied by a dramatic rise in NK activity, as well as an increase in the percentage of lymphocytes bearing the NK cell surface markers Leu 11a and Leu 19. Significant stimulation of NK activity was observed with beta-endorphin in vitro before exercise, but after exercise, beta-endorphin had a nonsignificant inhibitory effect. When these experiments were carried out in the presence of naloxone in vivo, the rise in NK activity after exercise was no longer significant. Naloxone did not significantly alter the rise in Leu 11a+ or Leu 19+ cell after exercise, as compared with the placebo. Finally, when naloxone was given to the subjects beforehand, exercise no longer completely blocked the in vitro beta-endorphin stimulation of NK cells. In conclusion, our observations that the exercise-induced augmentation of NK activity and the lack of effect of beta-endorphin in vitro on NK activity after exercise are both significantly attenuated by prior administration of naloxone suggest that the opioid system may play a major role in the modulation of NK cells during physiologic stress. PMID- 2972784 TI - Improved results following use of an advancement technique in the treatment of ileoanal anastomotic complications. AB - Between December 1981 and March, 1987, 188 ileal reservoir and ileoanal anastomosis procedures were performed at the University of Toronto. Anal anastomotic complications occurred in 24 (12.8%) patients. There were 19 patients who developed a leak as a result of non-healing of the ileoanal anastomosis (group 1). Late fistulae occurred in 5 patients (group 2). Various treatments were employed. In group 1, 2 of 15 (13%) patients treated with drainage and antibiotics, 3 of 7 (43%) in whom the anastomosis was resutured, and 5 of 7 (71%) in whom repair was performed by advancing the ileal outlet and resuturing the anastomosis, healed their anastomosis. One patient was treated successfully by performing a fistulotomy. Treatment was unsuccessful in one patient in whom the ileal reservoir and ileoanal anastomosis were revised and re-anastomosed. One patient (50%) in group 2 was treated successfully by advancing the ileal outlet and resuturing the anastomosis. Complete healing of the anastomosis was achieved in 12 patients (50%), although 3 patients subsequently required excision of the reservoir because of inability to evacuate spontaneously (1), incontinence (1), or psychological reasons (1). All nine (38%) patients who were functioning with their ileal reservoir were continent. One patient intubated the reservoir. The rate of urgency and soiling, and the number of bowel movements per 24 hours did not differ significantly from those of patients without these anastomotic complications. Our results suggest that early diagnosis and repair of ileoanal anastomotic dehiscence by advancement of the ileal outlet and resuturing of the ileoanal anastomosis results in improved reservoir salvage and satisfactory functional results. PMID- 2972786 TI - Longitudinal study of emotional, social, and physical changes after traumatic brain injury. PMID- 2972787 TI - Effects of retinoids on macrophage function and IL-1 activity. AB - The effects of three retinoids, all-trans-retinoic acid (RA), 13-cis-retinoic acid (cRA), and N-(4-hydroxyphenyl) retinamide (4-HPR), on macrophage function were evaluated. In vitro, RA, cRA, and 4-HPR caused a greater than twofold increase in phagocytosis of IgG-sensitized bovine erythrocytes (IgG-ORBC) by a mouse macrophage cell line (RAW). Significant increases in phagocytosis were produced by retinoid concentrations as low as 2 x 10(-10) M. RA also significantly increased phagocytosis of IgG-sensitized ORBC by BALB/c peritoneal macrophages in vitro. The ability of RAW macrophages to bind IgG-ORBC was significantly increased by 10(-6) to 10(-14) M RA. The potentiation of mitogenic responses of spleen cells to Con A and PWM by RA was relatively independent of macrophage function, i.e., splenocytes that were macrophage-depleted were responsive to the potentiating effects of RA. The effects of retinoids on T-cell dependent B-cell mitogenesis induced by PWM appeared not to be dependent on their previously reported capacity to alter prostaglandin synthesis. Treatment of spleen cells with 10(-6) M indomethacin did not abolish the potentiating effects of RA. However, RA in a dose-dependent fashion increased IL-1 activity at the level of the target T-cell. The greatest potentiation of IL-1 activity was at 10( 8) M RA. These results show that retinoids can modulate macrophage function at two different levels: potentiation of phagocytosis and potentiation of IL-1 activity at the level of the T-cell. PMID- 2972788 TI - Lipoprotein[a] is the major apoB-containing lipoprotein in the plasma of a hibernator, the hedgehog (Erinaceus europaeus). AB - We have undertaken studies aimed at elucidating the interrelationships existing between the seasonal modifications in endocrine status (already demonstrated by Saboureau, M., and J. Boissin. 1978. C.R. Acad. Sci. (Paris) 286D: 1479-1482) and plasma lipoprotein metabolism in the male hedgehog. During the course of these studies, we discovered that a lipoprotein comparable to human Lp[a] was a prominent component of the plasma lipoprotein spectrum in the hedgehog. This lipoprotein was present in the 1.040-1.100 g/ml density range (approximately), exhibited pre beta mobility upon agarose gel electrophoresis, and its Stokes diameter was 275 A. Its apolipoprotein moiety consisted of two proteins with molecular weights and amino acid compositions similar to those of human apoB-100 and apo[a], respectively. These two apolipoproteins were present in hedgehog Lp[a] as a complex that could be dissociated using dithiothreitol and whose stoichiometry could be 1:1. Lp[a] polymorphism due to size heterogeneity of apo[a] appeared to be present in the hedgehog as in man. The chemical composition of hedgehog Lp[a], obtained from animals bled during spring and summer, differed from that of its human counterpart in that the proportion of triglycerides was approximately three times higher in the hedgehog particle (13% vs. 4%), to the detriment of cholesteryl esters. Dissociation of the apoB:apo[a] complex has allowed us to obtain Lp[a] devoid of its specific polypeptide (Lp[a-]), a particle that retained the characteristics of Lp[a] as regards its lipid composition but whose Stokes diameter decreased by 30 to 40 A. The plasma concentration of LDL particles, defined as lipoproteins containing apoB-100 as their sole apolipoprotein constituent, was considerably lower than that of Lp[a]. These findings suggest that the hedgehog could be a unique animal model for studies regarding Lp[a] metabolism. PMID- 2972789 TI - Prosthodontic rehabilitation of the head and neck cancer patient. PMID- 2972790 TI - Venous thrombosis after abdominal surgery. A comparison between subcutaneous heparin and antithrombotic stockings, or both. AB - In an open controlled study, 248 consecutive patients (age more than 40 yrs) admitted for major abdominal surgery were randomized to one of three prophylactic antithrombotic treatments. Eighty-five patients received subcutaneous heparin, 74 patients had graduated compression stockings to the knee (TED stockings), and 89 patients had both subcutaneous heparin and stockings. Treatment began on the evening before operation and continued to complete mobilization, or for not less than five days postoperatively. On the fourth or fifth postoperative day, the patients underwent a 99mTc-plasmin test of the lower limbs as a test for deep vein thrombosis. There were 29.7% positive tests in the stocking group, 29.4% in the group with heparin prophylaxis, and 25.8% in the combined group. Differences between treatments were not statistically significant. PMID- 2972791 TI - Comparative effects of combined oral contraceptives containing desogestrel or levonorgestrel on serum lipids, lipoproteins and sex hormone binding globulin in healthy women. PMID- 2972792 TI - A new member of the immunoglobulin superfamily that has a cytoplasmic region homologous to the leukocyte common antigen. AB - A human gene (LAR) that hybridizes to mouse leukocyte common antigen cDNA under relaxed hybridization conditions was isolated. The LAR gene is expressed in a broad range of cells, including T lymphocytes, kidney, and prostate cells. The structure of the protein encoded by the LAR gene was deduced by determining the nucleotide sequences of a 7.7-kb LAR cDNA. The putative LAR protein is composed of a 1,234 amino acid extracellular region, a 24 amino acid transmembrane segment, and a 623 amino acid cytoplasmic region. The cytoplasmic region contains two homologous domains that have extensive sequence similarity to the cytoplasmic region of the leukocyte common antigens. The NH2-terminal region of the extracellular segment of the LAR protein contains three tandem Ig-like domains and nine non-Ig-like domains. Among the known Ig-like proteins, the LAR protein has the highest degree of similarity to neural-cell adhesion molecule. The non-Ig like domains of the LAR protein are also similar to the non-Ig-like domains of neural-cell adhesion molecule. PMID- 2972793 TI - Angioedema induced by a peptide derived from complement component C2. AB - Synthetic peptides that correspond to the COOH-terminal portion of C2b enhance vascular permeability in human and guinea pig skin. In human studies, 1 nmol of the most active peptide of 25-amino acid residues produced substantial local edema. A pentapeptide and a heptapeptide corresponding to the COOH-terminal sequence of C2b each induced contraction of estrous rat uterus in the micromole range; a peptide of 25 amino acids from this region induced a like contraction of rat uterus at a concentration 20-fold lower than the smaller peptides. The vascular permeability of guinea pig skin was enhanced by doses of these synthetic peptides in a similar fashion as that observed for the concentration of rat uterus. The induction of localized edema by intradermal injection in both the guinea pig and the human proceeds in the presence of antihistaminic drugs, suggesting that there is a histamine-independent component to the observed increase in vascular permeability. Cleavage of C2 with the enzymic subcomponent of C1, C1s, yields only C2a and C2b, and no small peptides, whereas cleavage of C2 with C1s and plasmin yields a set of small peptides. These plasmin-cleaved peptides are derived from the COOH terminus of C2b, and they induce the contraction of estrous rat uterus. PMID- 2972795 TI - Presentation of the H-Y antigen on Langerhans' cell-negative corneal grafts downregulates the cytotoxic T cell response and converts responder strain mice into phenotypic nonresponders. AB - We have used the murine cornea is an allograft model to investigate the relative roles of graft-derived IA+ APC (Langerhans' cells) and host-derived APC during the induction of CTL responses to H-Y. The natural exclusion of LC from the immunizing corneal graft led to a specific state of unresponsiveness to H-Y in responder strain mice, while inclusion of LC resulted in responsiveness. Failure to respond to H-Y could not be attributed to the absence of H-Y or IA antigen expression on the surface of LC-deficient grafts but instead, appeared to be due to active suppression of the T helper cell response during in vivo priming. Reprocessing of the H-Y antigen by host APC did not occur after immunization with H-Y presented on H-2-incompatible grafts unless presented initially by graft derived LC. H-2 as well as some non-H-2 alloantigens were presented to the host without a requirement for donor-derived LC. Thus there appear to be differential requirements for the processing and presentation of alloantigens. PMID- 2972794 TI - Identification of distinct C3b and C4b recognition sites in the human C3b/C4b receptor (CR1, CD35) by deletion mutagenesis. AB - Complementary DNA clones encoding the NH2-terminal region of human CR1 have been isolated and sequenced. The deduced complete amino acid sequence of the F allotype of human CR1 contains 2,039 residues, including a 41-residue signal peptide, an extracellular domain of 1,930 residues, a 25-amino acid transmembrane domain, and a 43-amino acid cytoplasmic region. The extracellular domain is composed exclusively of 30 short consensus repeats (SCRs), characteristic of the family of C3/C4-binding proteins. The 28 NH2-terminal SCRs are organized as four long homologous repeats (LHRs) of seven SCRs each. The newly sequenced LHR, LHR A, is 61% identical to LHR-B in the NH2-terminal two SCRs and greater than 99% identical in the COOH-terminal five SCRs. Eight cDNA clones were spliced to form a single construct, piABCD, that contained the entire CR1 coding sequence downstream of a cytomegalovirus promoter. COS cells transfected with piABCD transiently expressed recombinant CR1 that comigrated with the F allotype of erythrocyte CR1 on SDS-PAGE and that mediated rosette formation with sheep erythrocytes bearing C4b and C3b. Recombinant CR1 also had factor I-cofactor activity for cleavage of C3(ma). Analyses of six deletion mutants expressed in COS cells indicated that the NH2-terminal two SCRs of LHR-A contained a site determining C4 specificity and the NH2-terminal two SCRs of LHR-B and -C each had a site determining C3 specificity. The presence of these three distinct sites in CR1 may enable the receptor to interact multivalently with C4b/C3b and C3b/C3b complexes generated during activation of the classical and alternative pathways. PMID- 2972797 TI - Semantic facilitation with pictures and words. AB - The present experiments explored the role of processing level and strategic factors in cross-form (word-picture and picture-word) and within-form (picture picture and word-word) semantic facilitation. Previous studies have produced mixed results. The findings presented in this article indicate that semantic facilitation depends on the task and on the subjects' strategies. When the task required semantic processing of both picture and word targets (e.g., category verification), equivalent facilitation was obtained across all modality combinations. When the task required name processing (e.g., name verification, naming), facilitation was obtained for the picture targets. In contrast, with word targets, facilitation was obtained only when the situation emphasized semantic processing. The results are consistent with models that propose a common semantic representation for both picture and words but that also include assumptions regarding differential order of access to semantic and phonemic features for these stimulus modalities. PMID- 2972798 TI - Repetition, semantic priming, and stimulus quality: implications for the interactive-compensatory reading model. AB - Recognition latencies of single words were manipulated by repetition, degradation, or both, and the effects of context were observed. In both lexical decision and pronunciation tasks, repeated words were recognized faster than nonrepeated words yet were not any less affected by semantic context. Both inserting asterisks between a word's letters and masking slowed word recognition in comparison with a clear presentation, but only the masking manipulation showed contextual inhibition. In short, the magnitude of context effects did not always vary monotonically with the word recognition latencies in the neutral condition. Also, presentation of a word in an unrelated rather than related context did not produce larger repetition effects. The implications of these findings for Stanovich and West's (1983) and Jacoby's (1983) models are discussed. PMID- 2972796 TI - Interferon gamma induces synthesis of complement alternative pathway proteins by human endothelial cells in culture. AB - Human umbilical vein endothelial cells grown in vitro under standard conditions contain a high level of mRNA specific for the complement regulatory factors H and I. An additional 1.8-kb mRNA encoding a truncated form of factor H is also present. IFN-gamma stimulation of the cells causes a 6-7 fold increase in both factor H mRNA species, and a greater than 10-fold increase in factor I mRNA. IL-1 and LPS slightly suppressed factor H mRNA, while TNF had no effect. mRNA for factor B is also detectable in IFN-gamma-stimulated cells, but messengers for C1q, C4bp, and CR3 beta chain were not found. Secretion of factor H protein was also stimulated by IFN-gamma. The presence of mRNA for factors H, B, and I, together with C3 secretion, demonstrated by others, suggests that endothelial cells can assemble the complete alternative complement pathway. Endothelial cell complement may be involved in leukocyte-endothelium interactions mediated by leukocyte C3 receptors. PMID- 2972800 TI - Overflow, first-sight, and vanishing point distances in visual imagery. AB - The relationship between the size of a familiar object and the distances at which it is imaged is examined in three experiments. The distance at which an imaged object overflows the visual field is linearly related to object size, a result consistent with the size-distance invariance hypothesis (Kosslyn, 1980). The distance at which an object is initially imaged, first-sight distance, is related to the object size by a power function with an exponent less than 1. In addition, time required to scan from the first-sight to the overflow distance increases as a function of the difference between the two distance estimates. The distance at which an imaged object becomes too small to be identified, vanishing point distance, is related to object size by a power function with an exponent less than 1. This result does not support predictions made from the size-distance invariance hypothesis or Kosslyn's model of visual imagery. Implications for a theory of visual imagery and memory are discussed. PMID- 2972799 TI - Automatic and strategic processes in picture naming. AB - Prior presentation of related priming stimuli can facilitate responding to subsequent targets. However, recent research has demonstrated that inhibition effects can be produced by related primes when some of the primes in the testing sequence directly name their targets. This study used a picture-naming task, manipulating strength of relation between the prime word and the target picture, and the presence of identical primes in the testing sequence. Related primes facilitated naming when identical primes were absent, but not when they were present, whereas the strength effect did not vary as a function of identical prime presence. A second experiment replicated the strength effect and showed that it was not affected by the presence or absence of primary associates in the testing sequence. A comparison-strategy explanation of the inhibition effect is proposed, in which the strategy is superimposed on an automatic activation of related information. PMID- 2972801 TI - Imagery limitations in totally congenitally blind subjects. AB - Research on totally blind subjects performing tasks that involve visual imagery has often shown that they do not behave differently from matched sighted subjects, even when their blindness is congenital. If visual imagery is based on visual perception, such tasks may not required visual imagery. In the present article visual images are considered as representations maintaining some properties of visible objects and constructed on the basis of information from various sources. Owing to the absence of visual experience, the limitations of such representations are explored in a series of experiments requiring memorization of single nouns, pairs of nouns, or triplets of nouns associated with a cue noun. Recall by blind subjects was impaired when multiple interactive images (with noun pairs and triplets) are formed. The poorer recall of blind subjects reflected also loss of order information. Recall was better for both groups with locative noun cues and high-imagery targets. PMID- 2972802 TI - Musical imagery: generation of tones and chords. AB - Four experiments are reported that examine subjects' ability to form and use images of tones and chords. In Experiments 1 and 3, subjects heard a cue tone or chord and formed an image of a tone or chord one whole step in pitch above the cue. This image was then compared to a probe tone or chord that was either the same as the image in pitch, different from the image in pitch and harmonically closely related, or different and harmonically distantly related. Image formation times and response times were measured. In Experiment 3 a random-tone mask was used to control for possible contribution of the cue in echoic memory. In both experiments tone images were formed faster than chord images, a result consistent with the idea of structural complexity as a determinant of image formation time. Response times and accuracy rates were found to parallel results found in music perception studies, results consistent with the idea of shared mechanisms in the processing of musical images and percepts. Experiments 2 and 4 were control experiments examining the possible influence of demand characteristics and subjects' knowledge. In experiment 2 subjects predicted the results of "an average person" using imagery, and in Experiment 4 subjects completed the task without receiving imagery instructions. The findings ruled out the possibility that demand characteristics and subjects' knowledge were solely responsible for the results of Experiments 1 and 3 and support the argument for the role of imagery. PMID- 2972803 TI - Interference in spatial memory. AB - Four experiments examined interference in human spatial memory. The first two tested for retroactive interference by varying the similarity between the interpolated task and the task that was learned originally. Retroactive interference was observed during relearning of a radial stylus maze and during relearning of the locations of pairs of playing cards in a concentration game. In the third and fourth experiments, proactive interference was tested during the relearning of the locations of pairs in a concentration game. Some proactive interference was observed when the number of proactive games was varied (Experiment 3), and substantial proactive interference was found when the similarity between the proactive and target tasks was the independent variable (Experiment 4). Although extensive evidence indicates that spatial memory differs from other types of memory, spatial memory is subject to interference effects, as are verbal memory, motor memory, visual discriminative memory in animals, and animal spatial memory. PMID- 2972804 TI - Allocation of self-paced study time and the "labor-in-vain effect". AB - This research explored the possibility that a metacognitive control process (namely, the allocation of self-paced study time) might be affected by the output from metacognitive monitoring processes (i.e., ease-of-learning and/or feeling-of knowing judgments). In three experiments, university undergraduates received instructions that emphasized either accuracy of learning or speed of learning. The major findings were: (a) ease-of-learning judgments and feeling-of-knowing judgments are reliably related to study-time allocation, with more self-paced study time being allocated to the supposedly more difficult items; (b) even when instructed to master every item and when allowed unlimited study time to do so, people terminate study before learning is completed; and (c) large increases in self-paced study time can yield little or no increase in the subsequent likelihood of recall (the "labor-in-vain effect"). Implications are drawn for a model of the interplay between metacognitive monitoring processes and metacognitive control processes. PMID- 2972805 TI - Evidence for auditory temporal distinctiveness: modality effects in order and frequency judgments. AB - Two new, long-lasting phenomena involving modality of stimulus presentation are documented. In one series of experiments we investigated effects of modality of presentation on order judgments. Order judgments for auditory words were more accurate than order judgments for visual words at both the beginning and the end of lists, and the auditory advantage increased with the temporal separation of the successive items. A second series of experiments investigated effects of modality on estimates of presentation frequency. Frequency estimates of repeated auditory words exceeded frequency estimates of repeated visual words. The auditory advantage increased with frequency of presentation, and this advantage was not affected by the retention interval. These various effects were taken as support for a temporal coding assumption, that auditory presentation produces a more accurate encoding of time of presentation than does visual presentation. PMID- 2972806 TI - Data-driven and conceptually driven processes in partial-word identification and recognition. AB - Three experiments examined priming in partial-word identification (Warrington & Weiskrantz, 1968) and its relation to recognition memory. The results showed that changes in modality of presentation between study and test reduced performance on both identification and recognition. In contrast, changes in elaborative processing enhanced recognition but had no effect on identification. Furthermore, when explicit memory instructions were given, identification was changed to a cued recall test and was consequently affected by elaborative processing. We also found that the time course of forgetting in priming was different from that in recognition; priming in identification did not change over a 24-hr interval, whereas recognition declined rapidly during this interval. Overall, these results suggest that identification relies primarily on data-driven processing, whereas recognition can rely on both data-driven and conceptually driven processing. PMID- 2972808 TI - Long-term memory in amnesia: cued recall, recognition memory, and confidence ratings. AB - We explored what kind of information is acquired when amnesic patients are able to exhibit significant retention on tests of cued recall and recognition memory. Amnesic patients and control subjects attempted to learn sets of sentences. Memory for the last word in each sentence was tested after 1 hr in the case of the amnesic patients, or after 1 to 2 weeks in the case of (delayed) control subjects. Amnesic patients and (delayed) control subjects performed at similar levels on tests of cued recall and recognition memory. Amnesic patients were just as confident of their correct answers as were control subjects. However, amnesic patients were no more disadvantaged than control subjects when they were cued indirectly by presenting paraphrases of the original sentences. These findings demonstrate that the residual knowledge retained by amnesic patients can be as flexible, as accessible to indirect cues, and as available to awareness as the knowledge retained by (delayed) control subjects. PMID- 2972807 TI - More on recognition and recall in amnesics. AB - Hirst et al. (1986) reported that amnesic forced-choice recognition was relatively preserved when compared with amnesic recall. They equated normal recognition and amnesic recognition by extending exposure time for the amnesics and then comparing amnesic recall and normal recall. Amnesic recall was worse than normal recall, despite equated recognition. We conducted two experiments to extend that result. Experiment 1 established that the findings of Hirst et al. are not paradigm specific and hold when amnesic recognition and normal recognition are equated by increasing the retention interval for normals. In Experiment 2 we further established the generality of the result by examining yes no recognition. Findings further specify the selective nature of the direct memory deficit in amnesics. PMID- 2972810 TI - Why does back surgery have a bad name? PMID- 2972809 TI - Electrophysiological study of the effects of cardiodilatin 1-16 on the frog spinal cord in vitro. AB - Using the isolated spinal cord of the frog, hemisected and further divided into two distinct quadrants, we studied electrophysiological changes produced by peptides present in the atrial natriuretic factor (ANF) preprohormone. ANF and related peptides (atriopeptin I and atriopeptin III) did not affect the frog spinal cord. The 1-16 fragment from cardiodilatin (10(-5) M) induced slow depolarization in ventral and dorsal nerve stumps. The depolarization was associated with an increase of the evoked dorsal root potentials and depression of the fast component of the reflex responses. When depolarization approached its maximum value, spontaneous slow potentials appeared progressively similar to the evoked potentials, and became rhythmic until they reached a frequency of one potential every 15-20 seconds. The effects of cardiodilatin 1-16 are localized at dorsal horn level. It is suggested that this substance exerts a modulatory effect on frog cord physiology. PMID- 2972811 TI - The influence of incorporation of octadecenoic acid on the cell-associated fructosyltransferase and the extracellular glucosyltransferase activities of Streptococcus salivarius. AB - The rate of expression of the cell-associated fructosyltransferase (FTFm) activity of Streptococcus salivarius ATCC 25975 grown in continuous culture was linearly related to the rate of octadecenoic acid (C18:1) incorporation into the membrane lipids irrespective of the presence or absence of Tween 80 in the growth medium. This observation was confirmed with data obtained from cells grown in the presence of a series of n-alkanols. The results suggested that cosynthesis of lipids containing C18:1 residues was necessary for FTFm expression and accounted for the slight stimulation of enzyme expression by Tween 80 at all growth rates. In contrast, addition of Tween 80 to the growth medium resulted in several-fold increases in extracellular glucosyltransferase (GTFe) production irrespective of the growth rate. Following the addition of the surfactant to the growth medium, an exponential relation between the increased rate of GTFe production and the concomitant net increase in the rate of C18:1 incorporation was noted. The results obtained in continuous culture emphasized the underlying effect growth rate had on GTFe production, especially when Tween 80 was added to the growth medium. In the presence of n-alkanols, the rate of GTFe production plotted as a single 'U'-shaped curve with respect to the rate of C18:1 incorporation irrespective of the chain length of the n-alkanol studied. Rapid analyses of the extracellular proteins by SDS-PAGE suggested that hexan-1-ol and Tween 80 specifically stimulated the synthesis and secretion of GTFe and no other extracellular protein.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972812 TI - Molecular cloning and nucleotide sequence of the cyclomaltodextrin glucanotransferase gene from the alkalophilic Bacillus sp. strain no. 38-2. AB - The cyclomaltodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from the alkalophilic Bacillus sp. strain no. 38-2 was cloned in Escherichia coli using pBR322. A plasmid, pCS8, was isolated from a transformant producing CGTase and the cloned CGTase gene was found to be in a 5.3 kb DNA fragment. The nucleotide sequence of a 2.5 kb segment encoding the CGTase was determined. This segment showed an open reading frame which would encode a polypeptide of 712 amino acids. The pCS8 CGTase had the same enzymic properties as those of the extracellular CGTase produced by the alkalophilic Bacillus sp. strain no. 38-2. The nucleotide and amino acid sequences of this CGTase gene and gene product, respectively, have strong homology with those of the Bacillus macerans CGTase. PMID- 2972813 TI - Systemic lipopolysaccharide and pokeweed mitogen increase quinolinic acid content of mouse cerebral cortex. AB - The effects of lipopolysaccharide and pokeweed mitogen on brain L-tryptophan and quinolinic acid (QUIN) concentrations were investigated in C57BL/6NCR mice. Twenty-four hours after an intraperitoneal injection of lipopolysaccharide (5 micrograms from Salmonella abortus equii) or pokeweed mitogen (500 micrograms), cortical QUIN concentrations were increased by 81 +/- 6% and 182 +/- 15%, respectively. Plasma QUIN was increased 175 +/- 7% of control in pokeweed-mitogen treated mice only. Brain L-tryptophan concentrations were increased, whereas plasma L-tryptophan concentrations were decreased. The consequences of increased QUIN concentrations during endotoxin and mitogen exposure remain to be determined. PMID- 2972814 TI - Comparison of effects of D-1 and D-2 dopamine receptor agonists on neurons in the rat caudate putamen: an electrophysiological study. AB - Extracellular single-unit recording and microiontophoretic techniques were used to characterize the pharmacological properties of dopamine (DA) receptor subtypes within the rat caudate putamen (CPu), a striatal structure that receives a dense innervation from DA neurons originating from the substantia nigra pars compacta (A9 DA neurons). Similar to the action of DA, the DA D-1 receptor agonist (+)SKF 38393 generally potentiated the activation produced by glutamate (GLU) at low ejection currents (less than or equal to 5 nA); at higher ejection currents, it depressed 97% of the CPu neurons tested. By contrast, the D-2 receptor agonist LY 171555 (quinpirole) was much less effective in affecting the firing rate of CPu cells. The selective D-1 antagonist SCH-23390, administered either intravenously or iontophoretically, completely blocked the (+)SKF-38393-induced effects on CPu cells but failed to change the depressant effects produced by either quinpirole or 5-HT. On the other hand, the selective D-2 antagonist I-sulpiride, blocked the effects induced by quinpirole but not (+)SKF-38393. These observations suggest that the D-1 and D-2 DA receptor agonists elicit their effects via distinct DA receptor subtypes. A comparison of these results with our previous results obtained from the nucleus accumbens (NAc) indicates that NAc cells are more responsive to DA D-2 agonist, whereas CPu cells are more sensitive to D-1 agonist. Therefore, D-1 receptors in the CPu may have a critical role in mediating the effect produced by DA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2972815 TI - Demonstration of reperfusion after thrombolysis with technetium-99m isonitrile myocardial imaging. AB - Technetium-99m isonitrile myocardial perfusion imaging was employed in a patient undergoing thrombolytic therapy with recombinant tissue plasminogen activator for acute anteroseptal myocardial infarction. Technetium-99m isonitrile does not demonstrate significant myocardial redistribution after intravenous injection. The imaging agent was administered in the emergency room, prior to the initiation of thrombolytic therapy. The initial area at risk for infarction was visualized on images obtained after the patient had been effectively treated. Imaging performed 5 days later, after repeat injection of [99mTc]isonitrile, showed a smaller myocardial perfusion defect indicating salvage of myocardium. Thus, this technique offers promise as a noninvasive means of assessing the area at risk, the success of reperfusion, and the presence of salvaged myocardium, early in the course of acute myocardial infarction. PMID- 2972816 TI - Comparison of the clearance of technetium-99m MAG3 and iodine-131 OIH. PMID- 2972817 TI - [Studies on cochlear blood flow using laser Doppler flowmeter]. PMID- 2972818 TI - Hepatitis B vaccine in sickle-cell anemia. PMID- 2972819 TI - Ibuprofen: an inhibitor of alveolar bone resorption in beagles. PMID- 2972820 TI - Bacterial chemotactic peptide-degrading enzymes in human saliva. PMID- 2972822 TI - Spontaneous periodontitis in a sample group of the monkey Macaca fascicularis. PMID- 2972821 TI - Glomeruli in the molar gingival microvascular bed of germ-free rats. PMID- 2972824 TI - A contrast-correction method for digital subtraction radiography. PMID- 2972823 TI - Ultrastructural evidence of directed cell migration during initial cementoblast differentiation in root formation. PMID- 2972826 TI - Discrepancies that may arise if periodontitis is assessed on the basis of pocket depth alone. PMID- 2972825 TI - Proteases and inhibitors in the plasma of patients with periodontal disease. PMID- 2972827 TI - In situ lymphocyte subpopulations from active versus stable periodontal sites. AB - The purpose of this study was to evaluate lymphocyte subset densities and distributions within gingival biopsies from active sites (greater than or equal to 2 mm clinical attachment loss within three months of biopsy) versus clinically similar but stable or healthy sites. Small interproximal gingival biopsies representing at least one of each of the above categories were obtained from each of 20 periodontal maintenance patients. Serial cryostat sections displaying a cross section of the gingiva were labeled with monoclonal antibodies for (1) pan T cells, (2) T cytotoxic/suppressor cells, (3) T helper/inducer cells and (4) pan B cells and were developed using an avidin-biotin-peroxidase system. Lymphocyte populations were enumerated in repeatable fields from the sulcular, middle and oral one-third of each section. Relative proportions of the same lymphocyte subsets were analyzed in peripheral blood samples from the same patients using direct immunofluorescence. Pan B cells were significantly more prevalent in infiltrates from active sites than in stable (P less than 0.05) or healthy (P less than 0.01) sites. The T/B cell ratio was also significantly lower in active than stable biopsies (P less than 0.05), and in active biopsies versus blood (P less than 0.05). The T helper/T suppressor cell ratio did not vary significantly between blood and any gingival tissue disease group or location, but a trend toward lower relative numbers of T helper cells in the sulcular infiltrates of active sites was noted. These results support the premise that active periodontal sites display elevated B cell populations and abnormal immune regulation possibly involving the T helper cell subset. PMID- 2972828 TI - AIDS risk from needles less than 1 percent. PMID- 2972829 TI - Serotonin-induced reduction of the calcium-dependent plateau in frog dorsal root ganglion cells is blocked by serotonergic agents acting at 5-hydroxytryptamine1A sites. AB - Intracellular recordings from the dorsal root ganglion cells of adult frogs in the presence of tetraethylammonium display action potentials with a prominent calcium-dependent plateau. These action potentials can be altered by serotonergic agents in one of two ways. The superfusion of 5-HT (0.1-1 microM) usually produces a dose-dependent reduction of the action potential duration, whereas 8 hydroxy dipropylaminotetralin (8-OH-DPAT) (10-50 microM) produces a dose dependent increase in duration. A series of 5-HT antagonists were tested for their ability to block either the 5-HT or the 8-OH-DPAT effect. The antagonists were chosen for their reported selectivity in distinguishing receptors of the 5 HT1A, 5-HT2 and 5-HT3 subtypes. The antagonists' action on 5-HT narrowing [blockade by methiothepin, spiperone and spiroxitrine, but not by ketanserin or 3 tropyl-indole-3-carboxylate (ICS 205-930)] suggests that this response is mediated by 5-HT1A receptors. The widening effect produced by 8-OH-DPAT (a putative 5-HT1A agonist) was not blocked by any antagonist tested. At lower concentrations (0.1-2.5 microM) 8-OH-DPAT exhibited no agonist actions, but antagonized the 5-HT-induced narrowing. These results suggest the 5-HT receptors mediating 5-HT action potential narrowing in these cells are of the 5-HT1A subtype, but that they differ from the 5-HT1A receptors described in other tissues in which 8-OH-DPAT is an agonist or a partial agonist. PMID- 2972831 TI - Effects of time-limited vs unlimited compensation on pain behavior and treatment outcome in low back pain patients. AB - A common theme in the pain literature is that worker's compensation reinforces pain behavior and adversely influences treatment outcome of chronic pain patients. This study compared 110 chronic low back pain males divided into three groups: 44 receiving no compensation, 27 receiving time-limited worker's compensation, and 39 receiving unlimited social security disability benefits. All patients participated in a multimodal treatment program (e.g. nerve blocks, transcutaneous electrical nerve stimulation, relaxation training, biofeedback). Physician ratings of pain behavior and self-report measures of pain characteristics, activity level, and medication intake were gathered pretreatment; self-report measures were collected again approximately one year following treatment. The results showed disability patients to have a higher percentage of physician rated symptom dramatization and pain behavior and a greater usage of medication compared with the non-compensation and time-limited worker's compensation patients. At follow-up, no between group differences were found on measures of pain intensity, medication usage and activity. In general, however, more worker's compensation and non-compensation patients who were initially not working had returned to work at the time of follow-up compared with the disability patients. These results suggest that time-limited compensation may not affect treatment outcome or interfere with return-to-work chances while unlimited compensation may adversely influence the probability that patients will return to work. These findings support the notion that worker's compensation patients receiving time-limited financial benefits do not necessarily represent a 'problem' subgroup of chronic pain patients. PMID- 2972830 TI - Receptors with V1 characteristics mediate the maintenance of ethanol tolerance by vasopressin. AB - The neurohypophyseal hormone arginine vasopressin (AVP) acts in the central nervous system (CNS) to maintain functional tolerance to several effects of ethanol. The ability of exogenous vasopressin (administered i.c.v.) to maintain tolerance to the hypnotic effect of ethanol was blocked more effectively by antagonists acting at V1 receptors than by a V2-selective antagonist. Similarly, a V1-selective agonist was more potent than AVP in maintaining tolerance, whereas V2-selective agonists were inactive. These results indicate that vasopressin maintains ethanol tolerance via an action at CNS receptors that have the characteristics of V1 vasopressin receptors. Furthermore, a V1-selective antagonist, given alone, enhanced the rate of loss of tolerance, whereas a V2 selective antagonist did not, supporting a role for endogenous AVP, also acting at V1 vasopressin receptors, in the maintenance of ethanol tolerance. Characterization of the CNS receptors that mediate the modulation of ethanol tolerance by vasopressin suggests a mechanism of action of the peptide hormone in the CNS, and may contribute to the eventual development of therapies to modify functional ethanol tolerance. PMID- 2972832 TI - Seasonal endocrine cycles in the European hedgehog, Erinaceus europaeus. AB - Adult hedgehogs, maintained in captivity under natural environmental conditions of photoperiod and ambient temperature, were bled monthly. Plasma was assayed for melatonin, testosterone, prolactin, thyroxin, beta-endorphin and both plasma and urine for cortisol. Melatonin concentrations followed a circannual pattern, maximal between November and February at photoperiods less than 10L:14D, which is suggested as a key photoperiod. beta-Endorphin concentrations were maximal between March and September, coinciding with the reproductively active season of the hedgehog. Prolactin values were elevated during hibernation, indicating continued hypothalamo-pituitary axis activity. Testosterone and thyroxin levels were high between February and July and February and August respectively. During spring thyroxin concentrations rose 1 month later in females than males, reflecting the earlier arousal of the males from hibernation. There were no marked seasonal cycles of plasma or urinary cortisol. The results indicate photoperiod as the main factor in regulating hedgehog seasonality, with melatonin, beta-endorphin and prolactin important in the timing of reactivation of reproduction. Sexual differences in hedgehogs suggests environmental fine tuning of endogenous cycles, males being ready to inseminate females early in spring, while females only begin full breeding activity when conditions are suitable. PMID- 2972834 TI - The natural history of ankylosing spondylitis. PMID- 2972833 TI - Effect of cyproterone acetate on the pregnancy-blocking ability of male mice and the possible chemical nature of the pheromone. AB - Even though castration abolished the ability of alien males to induce implantation failure (the Bruce effect) in newly inseminated females, treatment of intact alien males with the steroidal antiandrogen, cyproterone acetate, for 14 days (short term) did not significantly depress their ability to induce the Bruce effect. However, prolonged treatment (42 days) with cyproterone acetate suppressed the pregnancy-blocking ability of alien males to some extent, possibly due to the antigonadotrophic properties of the drug. Ovariectomized alien females treated with implants of testosterone (androgenized females) exhibited the ability to block implantation in newly inseminated females, but concurrent treatment of androgenized females with cyproterone acetate did not depress this ability. The results strongly suggest that the pheromone involved in the male induced implantation failure is not the product of an androgen-dependent tissue, but is likely to be a product of androgen metabolism. PMID- 2972836 TI - Low back pain as a presentation of infective endocarditis in a patient with maladie de Roger. PMID- 2972835 TI - Further characterization of interleukin-2 production by lymphocytes of patients with systemic lupus erythematosus. AB - To further characterize the mechanisms responsible for defective interleukin-2 (IL-2) production in patients with systemic lupus erythematosus (SLE), we studied the effect of irradiation on the capacity of lymphocytes to produce this lymphokine when stimulated with phytohemagglutinin (PHA), or with a combination of PHA and a phorbol myristic acid ester (PMA). Irradiation increased PHA induced IL-2 production in patients with SLE and normal controls, and reached normal levels in 10 of 16 patients with SLE. This effect was due to inactivation of CD8+ suppressor cells. When PMA was used as a costimulant, maximal enhancement of IL-2 production was observed in both groups, but values in SLE remained significantly lower than in normals. These differences were not overcome by irradiation, raising the possibility that SLE suppressor cells act upon a site proximal to protein kinase C. Our studies have confirmed that active endogenous suppression may be responsible for most of the defective PHA induced IL-2 production in SLE and that this suppression is radiosensitive. PMID- 2972837 TI - Kuhse, Singer and slippery slopes. AB - Helga Kuhse and Peter Singer recently examined the view expressed by John Lorber that whereas at times it is permissible to allow severely handicapped infants to die, killing them must never be allowed. In attempting to demonstrate the mistaken nature of Lorber's fear that allowing active infanticide would lead us onto a slippery slope Kuhse and Singer make much use of John Harris's paper in the Journal of Medical Ethics in which he criticised Lorber's views. This paper examines some aspects of the case advanced by Kuhse and Singer and of the earlier paper by Harris. PMID- 2972838 TI - Anticipation in Huntington's disease is inherited through the male line but may originate in the female. AB - Data from the US National Huntington's Disease Roster have been analysed in terms of the difference in age of onset (AO) between affected parents and affected offspring, that is, in terms of 'anticipation'. While mean AO in offspring of affected mothers did not differ greatly from AO in their mothers, the distribution of AO in the offspring of affected fathers falls into two groups, the larger group showing an AO only slightly younger than their affected fathers and a small group whose AO was, on average, 24 years younger than their affected fathers. Analysis of the grandparental origin of the Huntington allele suggests that while propensity to anticipation is heritable for a number of generations through the male line, it originates at the time of differentiation of the germ line of a male who acquires the Huntington allele from his mother. It is suggested that major anticipation indicates an epigenetic change in methylation of the nucleic acid of the genome, which is imposed in the course of the 'genomic imprinting', that is, in the mechanism by which the parental origin of alleles is indicated. PMID- 2972841 TI - Enterotoxin B production and phage typing patterns of Staphylococcus aureus. PMID- 2972839 TI - Distinct mitogens reveal different mechanisms of suppressor activity in human cord blood. AB - We have studied the suppressive ability of human cord blood lymphoid cells in a three-days mixed lymphocyte culture proliferation assay stimulated by mitogen. Sex chromosomes served as markers for dividing cord (male) or maternal cells. Three distinct mitogenic agents were used in the co-cultures: the mitogenic lectin PHA, the anti-CD3 monoclonal antibody OKT3, and 12-0-tetradecanoyl-13 acetate (TPA), a direct activator of protein kinase C. With all mitogens we observed significant, non-specific suppression of maternal/adult cell division. However, three separate levels of suppression were evident. PHA-stimulated co cultures always showed the highest amount of cold suppressor activity (mean +/- SEM: 64.9 +/- 3.9). The mean suppression in OKT3- and TPA-stimulated co-cultures was 34.7 +/- 6.0 and 22.0 +/- 4.1%, respectively. Furthermore, indomethacin, a prostaglandin (PG) synthetase inhibitor, reduced by 41% the suppression in PHA driven co-cultures, whereas having no significant effect on the corresponding OKT3-driven co-cultures. Our results indicate the existence of an indomethacine sensitive, PG-dependent mechanism and a separate, indomethacine-resistant, mechanism of cord cell suppression. PMID- 2972840 TI - Mannose 6-phosphate receptors and their role in targeting proteins to lysosomes. PMID- 2972842 TI - c-myc products trans-activate the adenovirus E4 promoter in EC stem cells by using the same target sequence as E1A products. AB - Using a short-term transfection assay, we show that the E4 early adenovirus promoter is expressed to a certain extent in undifferentiated F9 and PCC4 cells, which are known to possess cellular E1A-like activity. We have also observed that c-myc products trans-activate the E4 promoter in EC stem cells and HeLa cells. Using 5' deletion mutants of the E4 promoter, we show that the same target sequence is used by c-myc and E1A. This sequence is located between positions 179 and -158 upstream of the cap site and is known to contain an activating transcription factor (ATF)-binding site. Moreover, the basal of level of activity of the deletion mutants. is related to the number of ATF binding sites. We therefore suggest that c-myc is a functional cellular homolog of the viral E1A gene and that it might correspond to one of the cellular E1A-like activities previously described for EC stem cells. We have also observed that only a c-myc plasmid coding for both p67 and 64 proteins, in contrast to one coding for p64 only, is able to trans-activate the E4 and E2A adenovirus promoter, suggesting that the p67 protein plays an essential part in activation. PMID- 2972844 TI - The P.A.D.U.A. (progressive augmentation by dilating the urethra anterior) procedure for the treatment of severe urethral hypoplasia. AB - Eight boys with congenital urethral hypoplasia and atresia are described, including 5 with a patent urachus, 2 with an "H" type urethral perineal duplication and 1 with a rectovesical fistula. Of the boys 5 had the prune belly syndrome. The technique of gradual progressive indwelling soft catheter or stent dilation achieved excellent results in 6 boys. This technique is believed to offer the slow stimulus necessary for urethral dilation that is missing in boys with a patient urachus or urethral duplication. The remaining 2 boys were treated with more conventional techniques of urethrotomy or rapid urethral dilation. They eventually achieved a satisfactory result but after significant morbidity. This clinical experience serves to describe and demonstrate how the application of gradual progressive soft, small catheter or stent dilation of severe urethral hypoplasia and atresia results in a functionally normal urethra with minimal morbidity. PMID- 2972843 TI - Regulation of adenovirus and cellular gene expression and of cellular transformation by the E1B-encoded 175-amino-acid protein. AB - Mutants of type 5 adenovirus that fail to express the E1B-gene-encoded 175-amino acid (175R) protein are unable to morphologically transform primary or continuous cultures of rat embryo fibroblast cells. This phenotype could result from a direct effect of this E1B polypeptide (along with E1A polypeptides) on cellular gene expression resulting in a pathway leading to altered cell growth or from an indirect role of the 175R protein made possible by its ability to modulate viral early-gene (most likely E1A) expression. To distinguish between these two models, viruses were constructed that expressed the individual E1A 13S and 12S genes in the presence of either the E1B 175R or 495R protein. Regardless of the E1A gene product that was expressed, viruses that failed to express the E1B 175R protein were transformation defective. Additional studies suggest that the E1A 289R protein and E1B 495R protein function in a common pathway leading to the establishment of the transformed cell. We also observe that E3 gene expression by viruses that fail to express the E1A 289R protein affects the efficiency of focus formation. When tested in both nonpermissive CREF cells and permissive HeLa cells, the lack of 175R protein expression appeared to have no effect (a transient twofold decrease in E1A mRNA accumulation was observed in CREF cells) on viral early-gene expression. These results suggest that the initiation of the transformed cell phenotype occurs because of some interaction in a common pathway between the viral E1A proteins and E1B 175R protein. Furthermore, we have shown that the E1B 175R protein does not enhance the rate of transcription initiation from the mouse immunoglobulin heavy chain gene promoter when these sequences are localized on a viral genome, and it does not diminish the ability of the E1A proteins to decrease the rate of enhancer-dependent transcription. PMID- 2972845 TI - Prevention of vascular prosthetic infection with an antibiotic-bonded Dacron graft. AB - Surfactant-mediated antibiotic bonding was used in an animal model of aortic prosthetic infection. Control grafts, control plus parenteral oxacillin, and oxacillin-bonded Dacron grafts were challenged by local inoculation with Staphylococcus aureus. Ninety percent of controls, 80% of parenteral antibiotic recipients, and only 30% of antibiotic-bonded Dacron grafts became infected (p less than 0.01, p less than 0.03). Antibiotic-bonded grafts were also superior in terms of suture line cultures and patency. In separate experiments in a subcutaneous pouch model, antibiotic bonding significantly improved the median infective dose of Dacron over that of controls and Dacron soaked in cephalosporin. These studies demonstrate that antibiotic-bonded Dacron implants are highly resistant to infection. A multicenter clinical trial is planned. PMID- 2972846 TI - A six-year follow-up of HIV-infected homosexual men with lymphadenopathy. Evidence for an increased risk for developing AIDS after the third year of lymphadenopathy. AB - Seventy-five homosexual men with generalized lymphadenopathy for at least three months (lymphadenopathy syndrome [LAS]), subsequently shown to be seropositive for antibody against human immunodeficiency virus, were enrolled in a prospective study in Atlanta in 1982 and 1983. As of Nov 30, 1987, twenty-two (29%) of the 75 were known to have developed acquired immunodeficiency syndrome (AIDS) three to 60 months after enrollment and five to 69 months after onset of LAS. The six-year cumulative incidence of AIDS, by Kaplan-Meier survival analysis, was 38%. The cumulative incidence in years 4, 5, and 6 (30%) was significantly higher than in years 1, 2, and 3 (11%), suggesting that the risk for AIDS increases after the third year of LAS and that many more study participants will eventually develop AIDS. A precipitous decline in the T-helper cell count frequently heralds the diagnosis of AIDS; this decrease appears to occur at different times after the onset of LAS in different persons. The four-year cumulative incidence of AIDS following observations of T-helper cell counts less than 200/mm3, 200 to 299/mm3, 300 to 399/mm3, and 400/mm3 or greater was 84%, 41%, 25%, and 18%, respectively; these data are important for determining prognosis in the individual patient as well as for determining the suitability of candidates and baseline data for drug trials. PMID- 2972847 TI - Leads from the MMWR. Multistate outbreak of sporotrichosis in seedling handlers, 1988. PMID- 2972848 TI - Immediate vs delayed catheterization and angioplasty following thrombolytic therapy for acute myocardial infarction. TIMI II A results. The TIMI Research Group. AB - The Thrombolysis in Myocardial Infarction II A Study investigated whether immediate cardiac catheterization with percutaneous transluminal coronary angioplasty (PTCA), when appropriate, would confer an advantage over the same procedures performed 18 to 48 hours later. All patients were treated with intravenous recombinant tissue-type plasminogen activator within four hours of the onset of acute myocardial infarction. Percutaneous transluminal coronary angioplasty of the infarct-related artery was attempted in 72% of the 195 patients assigned to immediate PTCA; 84% of the attempts were judged to have shown improvement. Percutaneous transluminal coronary angioplasty was attempted in 55% of the 194 patients assigned to 18- to 48-hour PTCA; 93% of the attempts were judged to have shown improvement. No differences between the two PTCA groups were observed for ejection fraction (primary end point), measured by contrast ventriculography predischarge (50.3% in the immediate and 49.0% in the delayed PTCA groups). Immediate catheterization/angioplasty was associated with increased frequency of bleeding and coronary artery bypass surgery. These findings indicate that immediate performance of coronary arteriography and PTCA compared with delaying these procedures for 18 to 48 hours provides no advantage and may be harmful. PMID- 2972849 TI - The aggressive war on acute myocardial infarction: is the blitzkrieg strategy changing? PMID- 2972850 TI - RNA splicing and genes. AB - The splicing of long transcripts of RNA (copied from DNA in the cell nucleus) into smaller, specific mRNA (ready for export to the protein-producing machinery in the cytoplasm) is an important event in the regulation of gene expression in eukaryotic cells. The splicing reaction occurs as a late step in the nuclear pathway for synthesis of mRNAs. This pathway commences with initiation of transcription by RNA polymerase II and probably involves an integrated series of steps each dependent on previous events. Splicing of precursors to mRNAs involves the formation of a spliceosome complex containing the 5' and 3' splice sites. This complex contains the evolutionarily highly conserved small nuclear RNAs (snRNAs) U2, U4, U5, and U6. The most abundant snRNA, U1, is required to form the spliceosome and may be a part of the spliceosome. Analogues of these snRNAs have been identified in yeast. Assembly of the spliceosome probably involves the binding of a multi-snRNA complex containing U4, U5, and U6 snRNAs. Several observations suggest that the association of snRNAs in such complexes is quite dynamic. It is argued that the snRNAs in the spliceosome form a catalytic RNA structure that is responsible for the cleavage and ligation steps during splicing. PMID- 2972851 TI - Treatment of human immunodeficiency virus-related thrombocytopenia with zidovudine. AB - Human immunodeficiency virus (HIV)-related thrombocytopenia has been well described and requires therapy in about one half of the patients. Conventional modes of therapy with prednisone, danazol, immunoglobulin, and/or splenectomy have not been uniformly successful. We have administered zidovudine to three patients with HIV-related thrombocytopenia. All three patients responded with a sustained increase in their platelet counts, despite discontinuation of conventional therapy. Interruption of zidovudine therapy was associated with a decrease in platelet count. Concomitant with the elevation in platelet count with zidovudine therapy, there was a reduction in the circulating p24 antigen levels. Whether the elevations in the platelet count in these patients with HIV-related thrombocytopenia is due to the antiviral effect of zidovudine is unknown. It is clear that further studies examining the prospective use of zidovudine in the treatment of HIV-related thrombocytopenia are indicated. PMID- 2972852 TI - Degrading process of acute ischemic myocardial cells. AB - 1. Development of acute ischemic myocardial injury was studied in mongrel dogs, induced by ligation of left anterior descending coronary artery (LAD), by biochemical analysis of myocardial fractions such as sarcoplasmic reticulum (SR) and mitochondria (Mt) and by electron microscopic observation of ischemic myocardial cells with lanthanum probe method. 2. Irreversible injury of ischemic myocardium initiated in subendocardial muscle as early as 20 min after occlusion of LAD as expressed degradation of major ATPase protein and phosphatidylcholine and phosphatidylethanolamine of SR and irreversible impairment of state III respiratory and dinitrophenol (DNP)-ATPase activities of Mt, and these necrotic changes advanced to subepicardial layer at about 60 min. 3. Ultrastructural irreversible findings appeared later at about 60 min following inflow of lanthanum ions in ischemia for 30 min. 4. Activation of cathepsin B inside of SR under ischemic acidic metabolism and abnormal inflow of Ca++ into ischemic cardiac myocytes are suspective of very important factors for the initiation of myocardial ischemic injury in early myocardial ischemia. PMID- 2972853 TI - Reocclusion of the infarcted vessel during acute interventions in myocardial infarction. AB - Acute reocclusion of the recanalized infarcted vessel was observed during thrombolytic therapy in 28 or 138 patients, and during angioplasty following thrombolysis in 5 of 70 patients. The occluded vessel reopened again in 20 of 28 patients by additional administration in nitroglycerin and urokinase and in 3 of 5 patients by repeated angioplasty. There was no significant difference in success rate of thrombolysis or angioplasty between patients with acute reocclusion and those without. However, reinfarction after acute intervention tended to be more common in the former patients than the latters. PMID- 2972854 TI - [Effect of alpha-human atrial natriuretic polypeptide on renal, cardiovascular and endocrine functions during haemorrhagic shock in dogs]. PMID- 2972855 TI - Hematocrit increase by mental stress in hypertensive patients. AB - Hematocrit (Hct) changes during mental stress were studied in 21 patients with mild hypertension. A 10 min arithmetic stress test increased blood pressure from 129 +/- 2/80 +/- 2 mmHg (mean +/- SEM) to 167 +/- 5/98 +/- 3 mmHg, and heart rate from 64 +/- 2 to 81 +/- 3 (p less than 0.001). Concomitantly, a significant (p less than 0.001) increase in Hct was observed (from 41.5 +/- 0.7% to 42.2 +/- 0.8%), and increments of Hct (-0.1% to 2.2%) were significantly (r = 0.61, p less than 0.01) correlated with those of systolic blood pressure. Plasma levels of norepinephrine (NE), epinephrine (E) and atrial natriuretic peptide (ANP) also showed a significant increase after mental stress. All these changes disappeared after a 60 min recovery period. The results suggest that mental stress increases Hct promptly, which may be due to concomitant increases in plasma NE, E and ANP levels. Hct elevation resulting from mental stress may be clinically relevant, at least, in hypertensive patients who are at risk of occlusive vascular diseases. PMID- 2972856 TI - Transient platelet aggregation as a mechanism of unstable angina. Aggressive treatment with coronary angioplasty. AB - Decreased oxygen supply rather than increased demand may be the primary pathogenic mechanism of myocardial ischemia in patients with unstable angina. Coronary artery spasm and in vivo platelet aggregation may play an important role in the mechanism by which the magnitude of fixed obstruction secondary to coronary atherosclerosis is transiently exacerbated. In this case report, we describe a patient who developed chest pain accompanied by ischemic ECG changes during coronary arteriography, due to a transient thrombotic aggravation of a fixed, significant coronary stenosis. Percutaneous transluminal coronary angioplasty was performed with relief of the coronary stenosis and of the symptoms. PMID- 2972857 TI - Distribution of hematoporphyrin derivative in normal and malignant tissue. AB - The distribution of [14C]hematoporphyrin derivative (HpD) has been studied in mice bearing human lung tumor and in guinea pig bearing Line 10 tumor. The amount of [14C]HpD in the transplanted tumor tissue of mice at various times following drug administration (20 mg/kg) was higher than in skin or muscle tissue but was less than in liver, kidney, spleen, lung and heart tissue. The distribution pattern of [14C]HpD in guinea pig at 2 days following injection was also similar to that in mice. Our data were different from those that suggested the selectivity of uptake and retention of HpD by malignant tissues, using fluorometric techniques. PMID- 2972858 TI - [A case of early carcinoma of the gallbladder--usefulness of cytological diagnosis in the bile in early diagnosis under laparoscopy]. PMID- 2972859 TI - Nephritogenicity of proteoglycans. II. A model of immune complex nephritis. AB - Antibodies to glomerular basement membrane, heparan sulfate-proteoglycans are nephrotoxic but possess a weak nephritogenic potential. In order to enhance the nephritogenic potential, the antibodies were intravenously administered into rats presensitized with heterologous rabbit IgG. This resulted in the integration of heterologous and autologous phases, the two phases characteristic of the traditional model of nephrotoxic serum nephritis. The presensitization caused a dramatic shift in the binding characteristics of the heterologous antibodies between the kidney and lymphoid tissues. A proliferative form of immune complex glomerulonephritis associated with a remarkable proteinuric response was observed. In addition, a moderate degree of hematuria was noted as well. The proteinuria was largely complement-dependent and may possibly be cell-mediated as well. The proteinuria became severe with increasing production of host IgG antibodies and with their subsequent sequestration in the glomeruli. The predominant glomerular lesions were in the form of epimembranous/subepithelial immune deposits, which became more frequent with timely increasing titer of host autologous IgG antibodies. These findings indicate that antibodies to heparan sulfate-proteoglycan, an authentic component of the basement membrane, are capable of mediating a glomerular injury with acquisition of nephritogenic potential in an appropriate environment of the host. At present, it seems that this is the sole constituent of the basement membrane whose antibodies are capable of inducing an immune complex nephritis. PMID- 2972860 TI - Nephritogenicity of proteoglycans. III. Mechanism of immune deposit formation. AB - Administration of antibody, directed against glomerular basement membrane (GBM) heparan sulfate-proteoglycan, into a presensitized rat results in the induction of membranous nephropathy with subepithelial immune-complex deposits. In this investigation, we examined the mechanisms responsible for the formation of subepithelial immune-complex deposits in the anti-HS-PG model. In initial experiments, the intravenously administered radioiodinated antibody was seen exclusively localized in the regions of the glomerular capillary wall where the subepithelial deposits were observed. To determine their exclusive localization in the subepithelial space, kinetics of movement of the intravenously administered antibody was investigated. The antibody localized in the inner layers of the GBM within a few minutes after its administration. It equilibrated in the inner and outer layers of the GBM in a matter of a few hours. Then, after 24 hours, it gradually disappeared from the inner layers of the GBM and persisted in the outer layers only. The ready clearance of the antibody from the inner layers may be related to the differential in the kinetics of lateral intrinsic plasma fluid currents within the GBM. The persistence of heterologous antibody exclusively in the outer layers and the availability of host autologous antibodies probably resulted in the development of immune complex deposits in the subepithelial space. The glomeruli devoid of plasma water currents showed no change in the concentration of the antibody in the inner and outer layers of the GBM or mesangial matrix. Also, no antibody binding was observed with the plasmalemma of either the foot processes or visceral epithelia. The data suggest that the biochemical-biophysical properties of the glomerular capillary wall, in concert with its intraglomerular hemodynamics, most likely played a significant role in the development of subepithelial immune-complex deposits in this model. PMID- 2972861 TI - Atrial natriuretic peptide in gentamicin-induced acute renal failure. PMID- 2972862 TI - The effect of atrial natriuretic factor on blood pressure and renal function of long-term hypothyroid rats. PMID- 2972863 TI - Hypertension in renal transplantation. PMID- 2972864 TI - Effect of converting enzyme inhibitors on cardiac changes in experimental uremia. PMID- 2972865 TI - Atrial natriuretic hormone: a regulator of blood pressure and volume homeostasis. AB - An explosion of research over the last seven years has led to the discovery and characterization of a peptide, originating in the heart's atria, that possesses impressive vasorelaxant and natriuretic properties. Although the several atrial peptides that have been isolated by researchers working in different laboratories vary in length, all contain the same core sequence. Cardionatrin I, a 28-amino acid peptide, is the likely active circulating hormone. In the atrial peptide's action on vascular smooth muscle, it appears especially to counter the effects of angiotensin II. The peptide's effects on renal hemodynamics and sodium excretion include a marked increase in glomerular filtration rate. Atrial hormone also induces impressive natriuresis in experimental animals, for which an increase in glomerular filtration rate may be a necessary component. Atrial hormone has been found to reduce arterial blood pressure in both animals and humans. In experimental animals, the peptide appears to lower blood pressure by different mechanisms in high- and low-renin forms of hypertension, and to lower pressure to a greater degree and with lower doses in the former as compared with the latter. In patients with essential hypertension, primary aldosteronism, congestive heart failure, renal failure, and perhaps ascitic cirrhosis, plasma ANH levels tend to be higher than they are in normotensive individuals. Atrial hormone causes marked and sustained suppression of renal renin secretion and, thus, of plasma renin levels. In addition, atrial hormone blocks aldosterone secretion and opposes the vasoconstrictive effects of angiotensin II and the sodium-retaining action of aldosterone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972866 TI - Renal response to low-dose infusion of atrial natriuretic peptide in normal man. PMID- 2972867 TI - Uranyl nitrate-induced acute renal failure in rats: effect of atrial natriuretic peptide on renal function. PMID- 2972868 TI - Acute hemodynamic effects of atrial natriuretic peptide in essential hypertension. PMID- 2972869 TI - Plasma levels of atrial natriuretic peptide in chronically dialyzed patients. PMID- 2972870 TI - Is the effect of calcium diet or parathyroidectomy on the development of hypertension in spontaneously hypertensive rats mediated by atrial natriuretic peptide? PMID- 2972871 TI - Mechanism of action of atrial natriuretic peptides does not involve adenylate cyclase system of different rat brain areas. PMID- 2972872 TI - [Organization of a surgical service for children in a district hospital]. PMID- 2972873 TI - [Effect of muscle relaxants on intraocular pressure]. AB - Since general anesthesia is being used increasingly in ophthalmic surgery, the effects of anesthetic drugs on intraocular pressure (IOP) have to be considered. Competitive neuromuscular blocking drugs either do not affect IOP or produce a slight decrease. Depolarizing muscle relaxants increase IOP. This effect, which is pronounced with succinylcholine, cannot be reliably prevented by any concomitant medication. The new competitive relaxants atracurium and vecuronium provide stable conditions with respect to IOP and systemic circulation, combined with fast onset and intermediate duration of action. PMID- 2972875 TI - Atrial natriuretic peptide in human urine. AB - A highly sensitive radioimmunoassay to measure atrial natriuretic peptide (ANP) concentration in urine has been established, and its clinical usefulness is presented. ANP in urine was stable at 4 degrees C for several days and was easily measured by our radioimmunoassay. The average ANP excretion in 65 healthy persons was 25.0 +/- 1.4 ng/day (mean +/- SEM) and the fractional excretion of ANP was 0.7 +/- 0.05%. In 14 patients with congestive heart failure, the average ANP excretion was 119.2 +/- 29.4 ng/day, which decreased to 53.3 +/- 11.0 after successful treatment. PMID- 2972874 TI - Isolation and structural analysis of "urodilatin", a new peptide of the cardiodilatin-(ANP)-family, extracted from human urine. AB - Two major forms of cardiac peptides have been established in the last few years: (a) a prohormone of 126 amino acids (CDD/ANP-1-126) in the endocrine heart and (b) the circulating CDD/ANP-99-126 (= alpha ANP) in blood plasma. The method we applied earlier to isolate the circulating form of cardiodilatin from human blood was used to detect and analyze the biologically active, predominant form of the same polypeptide family excreted by the kidneys. Each step of the isolation procedure was followed up by a bioassay using an in vitro vascular smooth muscle relaxation test and a highly specific RIA against cardiodilatin (CDD-99-126) for the initial purification steps. The polypeptides excreted in 1000 l of normal human urine were adsorbed to 2.5 kg of alginic acid, and after elution and lyophilization processed on a G-25 Sephadex column. The obtained crude polypeptide fractions were applied to ion-exchange chromatography. Thereafter four steps of HPLC were carried out to purify the polypeptide which was the suggested form of cardiodilatin (CDD) in human urine. The amino acid analysis and gas phase sequence analysis showed that the main form of urinary cardiodilatin is a 32 amino acid residue containing molecule, cardiodilatin-95-126. The molecule is N-terminally extended compared to the circulating CDD-99-126. This suggests that the analyzed urinary peptide is not the residual plasma form, filtrated and renally cleared from blood, but probably a polypeptide produced and processed in the kidney tubules and cleaved by a different postranslational process. Therefore, this vasorelaxant polypeptide is called urodilatin. PMID- 2972876 TI - [Secondary immune deficiency in renal failure exemplified by hepatitis B vaccination]. AB - In dialysis patients the immune response to hepatitis B-vaccination is greatly impaired. In vitro the non-responders show a failure of the monocytes to support the process of primary T-cell activation. This defect results in a lack of interleukin 2-production and an enhanced sensitivity of the interleukin-2 receptor system. Addition of low doses of interleukin-2 fully reconstitutes the deficient immune response in vitro. Furthermore, the local application of low dose interleukin-2 during a standard vaccination with 40 micrograms hepatitis B vaccine normalizes the non-responder state in vivo. PMID- 2972878 TI - Sympathoadrenal activity and sympathoinhibitory hormones during acute and chronic nicotine application in dogs. AB - Though acute nicotine administration results in increased blood pressure and heart rate, previous work has shown that chronic nicotine treatment does not result in significant hypertension. In fact, surprisingly it has been shown to produce hypotension. We performed the present experiments to further analyze the effects of chronic nicotine treatment. In untreated dogs (n = 7) under pentobarbital anesthesia (with adrenal hormone release measured directly by cannulation of the adrenolumbar veins) cumulative nicotine infusions (1-24 micrograms/kg/min i.v.) caused dose-dependent release of epinephrine (from 3.0 +/ 0.7 to 111 +/- 30 micrograms/kg/min) and norepinephrine (from 0.4 +/- 0.1 to 11.2 +/- 3.1). However, significant release of leu-enkephalin and met-enkephalin immunoreactivity was observed only with the highest nicotine infusion (24 micrograms/kg/min). In untreated conscious dogs (n = 12), nicotine test infusions (3 and 10 micrograms/kg/min), 15 min, yielded smoking relevant plasma nicotine levels and augmented heart rate, mean arterial pressure, plasma catecholamine levels, and adrenal epinephrine release. Plasma-enkephalin immunoreactivities were only marginally elevated with the higher nicotine test infusion. Chronic nicotine treatment (1.5 micrograms/kg/min s.c. for 5 weeks, n = 7), only transiently (first 1-2 weeks) augmented mean arterial pressure, heart rate, and epinephrine release, but during the plateau phase of treatment, hemodynamics and catecholamine parameters were identical to the pretreatment period. Acute responses of hemodynamics and catecholamines to nicotine test infusions declined progressively during chronic treatment, but the time course of this attenuation seemed not related to the reversal of the transient hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972880 TI - [Clinical variants of the neurodystrophic syndrome of the muscles of the anterior abdominal wall in osteochondrosis of the spine]. PMID- 2972879 TI - [Ultrasonic study of the intestines]. PMID- 2972881 TI - A technique for creating a porcine end colostomy. PMID- 2972882 TI - Branching enzyme in erythrocytes. Detection of type IV glycogenosis homozygotes and heterozygotes. PMID- 2972877 TI - Pathobiochemical aspects of diabetic nephropathy. AB - Diabetic nephropathy develops in many diabetic patients as consequence of glomerulosclerosis. On the basis of a series of recent observations it is suggested that a combination of metabolic and hemodynamic changes is responsible for the pathogenesis of diabetic nephropathy. Since the glomerular filtration unit has been characterized to consist of collagen type IV and minor components like laminin, fibronectin and heparan sulfate proteoglycan, influence of diabetes on basement membrane (BM) components has been studied. Biochemical alterations of glomerular BM consist of an increased nonenzymatic glucosylation of type IV collagen leading to unphysiological crosslinking. This, in turn, may result in alteration of the size selective properties of the glomerular filtration unit. Changes in composition of glomerular BM have recently been described. An increased synthesis of type IV collagen with concomitant decrease of heparan sulfate proteoglycan may lead to alteration of the charge selective barrier and consequently to increased permeability of the glomerular BM. Permanently unbalanced synthesis of BM components finally results in obliteration of the capillary lumen. In late state nephropathy intrinsic basement membrane components are no longer produced. Instead, massive accumulation of PAS positive material occurs. PMID- 2972883 TI - Ultrastructure of the macula densa in streptozotocin diabetic rats. AB - Morphometric analyses of the macula densa in streptozotocin diabetic rats have revealed, that the volume density of the large lateral intercellular spaces, which are present in normal animals between the macula densa cells, decreases significantly in magnitude from 8.7 to 1.5% in diabetic animals. The volume density of cytoplasmic glycogen in the macula densa cells increases significantly from 5.4% in the controls to 14.8% in the diabetic animals, but the total volume density of mitochondria is the same in the two groups. The contact area between macula densa cells and the Goorgmaghtigh cells is increased by 38% in the diabetic animals compared with controls. The structural abnormalities in the macula densa in response to diabetes might be considered a structural counterpart to the alterations in the tubuloglomerular feedback mechanism and the increase in glomerular filtration rate in diabetes. PMID- 2972885 TI - Ingroup and outgroup evaluation by disabled individuals. PMID- 2972884 TI - South Carolina florists dermatitis: case report and survey results. PMID- 2972886 TI - Long-term treatment of severe dysarthria: a case study. AB - This case study describes the long-term treatment and changing symptoms in a single subject with dysarthria secondary to basilar artery thrombosis. Initially, the subject was anarthric. Treatment efforts thereafter were directed toward modifying speech respiration, velopharyngeal function, articulatory precision, speech intensity, and speech intelligibility. A variety of treatment and measurement techniques are illustrated. The behavioral change resulting from each of the treatments was small. However, when combined, these small gains in conjunction with some neurological recovery resulted in significantly improved communication and quality of life for this subject. Implications for management of similar subjects are discussed. PMID- 2972887 TI - The effect of photodynamic therapy on the microcirculation. AB - Photodynamic therapy (PDT) is a new form of cancer therapy involving tumor localization by photosensitizing drugs such as dihematoporphyrin ether (DHE). Light irradiation of drug-sensitized tissue results in photoactivation of DHE and tumor necrosis. The mechanism of action is incompletely understood but involves the generation of singlet oxygen which produces cytotoxic effects on tissues containing the compound. In addition, microcirculatory aberrations have been described during PDT. We have studied the acute effects of PDT on the microcirculation using in vivo television microscopy of the rat cremaster. This has enabled us to observe the effects of PDT on both paired and unpaired arterioles and venules using two different wavelengths of activating light (blue, 450-490 nm, and green, 530-560 nm). For both wavelengths of activating light, significant reduction in blood flow of all vessels was seen during PDT. This, in combination with the formation and embolization of platelet thrombi, resulted in stasis of blood flow in 80% of arterioles and 90% of venules. Observation for 2 hr after the completion of photoactivation revealed reperfusion in 20% of the arterioles but none of the venules. Blood flow was reduced by a combination of vasoconstriction and platelet thrombus formation. Reducing the total activating energy from 120J/cm2 to 24J/cm2 significantly reduced the response in venules and the incidence of stasis in both arterioles and venules. We conclude that the photoactivation of DHE results in significant vasoconstriction and thrombosis of normal vessels and that if these effects are seen at later times after DHE administration and in tumor neovasculature they may contribute to the mechanism by which PDT results in tumor necrosis. PMID- 2972888 TI - Arginine, protein malnutrition, and cancer. AB - The amino acid arginine has anabolic and immunostimulatory properties. This study evaluated the potency of arginine in limiting the severe nutritional and immunological insults of protein calorie malnutrition and increasing tumor load. In protein-depleted A/J mice (n = 340) bearing either an immunogenic (C1300) or poorly immunogenic (TBJ) neuroblastoma, arginine supplementation [1%] significantly augmented T lymphocyte responses (mitogenesis, interleukin-2 production) compared with both a glycine-supplemented and nonsupplemented group. Arginine supplementation significantly retarded the growth of C1300 and prolonged median host survival. These results correlated with augmented autologous mixed lymphocyte tumor cell responses and enhanced specific cytotoxicity. This anti tumor effect was not demonstrated in mice bearing TBJ where both arginine and glycine stimulated tumor growth compared with nonsupplemented mice. There was no significant difference between arginine and glycine in preservation of carcass weight. These studies suggest that the immunostimulatory effects of arginine are not due to supplemental nitrogen and that an associated antitumor effect is dependent on tumor antigenicity. PMID- 2972889 TI - T cell receptor gene rearrangements in B-precursor acute lymphoblastic leukemia correlate with age and the stage of B cell differentiation. AB - Children with ALL diagnosed at less than 2 years of age have a poor prognosis when compared with older children. In an effort to identify biologic features of ALL in children less than 2 that might explain this difference, we performed extensive immunophenotypic and molecular genetic analyses on a series of patients. For comparison purposes patients were divided into four groups: CALLA- (CD10-) infants less than 2 years of age at diagnosis (n = 10), CALLA- children greater than 2 years of age at diagnosis (n = 10), CALLA+ infants (less than 2 years, n = 21) and CALLA+ children (older than 2 years, n = 21). No immunophenotyping differences in CALLA- or CALLA+ subgroups were identified when cases less than 2 were compared with cases greater than 2 years of age at diagnosis. The most interesting results were in the CALLA- group where 94% of the samples expressed the B cell antigen CD19 but 27% co-expressed CD7. Double labeling experiments confirmed leukemic blast cells co-expressed CD19 and CD7. The double-labeled cells represent either leukemic conversion of a precursor cell which has not yet committed to B or T cell lineage or aberrant expression of these antigens. Molecular genetic studies demonstrated that all samples, regardless of the patients' age or immunophenotype, had rearrangement of the Ig heavy chain gene. The most striking molecular results were in CALLA- patients; in patients less than 2 at diagnosis neither the beta- nor the gamma-chain gene of the T cell receptor (TCR) was rearranged, whereas DNA from 5 of 10 patients over the age of 2 demonstrated beta- or gamma-chain TCR gene rearrangements. The percentage of CALLA+ cases under the age of 2 years with rearrangements in TCR genes is less than that found in CALLA+ cases over the age of 2 years. The finding of no TCR rearrangements in CALLA- ALL and a decreased number of gamma TCR rearrangements in CALLA+ cases under the age of 2 suggest that age may affect TCR gene rearrangements in lymphoblasts. The molecular differences in TCR gene rearrangements do not appear to correlate with the response to therapy. PMID- 2972891 TI - Characteristics of the recurrently hospitalized adult. An information synthesis. AB - Hospital use in the US is concentrated in the approximately 2% of the population that is repeatedly hospitalized. Using the research integration technique of information synthesis, empirical studies of readmission were assembled and analyzed to determine the demographic and clinical characteristics of the repeatedly hospitalized adult. The authors' synthesis of articles that met preset criteria of relevance and validity indicated that clinical variables show a more consistent association with readmission than demographic variables. Diagnosis, prior use, and disability appear to be strong predictors of an individual's level of hospital use. Demographic variables with consistent positive associations with readmission include being widowed, living with relatives other than a spouse, decreasing income, living in the South, and living in an urban area, but available work does not indicate whether these demographic variables are independent predictors of readmission or merely markers of an increased prevalence of chronic disease. Findings indicate that future investigations of the predictors of this costly pattern of hospital use will be most illuminating if illness-related factors such as diagnosis, disease severity, functional status, and usage history are carefully specified and examined. PMID- 2972892 TI - [Vaccination against hepatitis B. Follow-up with antibody tests]. PMID- 2972890 TI - Adult T cell leukemia cell lines that originated from primary leukemic clones also had a defect of expression of CD3-T cell receptor complex. AB - Surface phenotypes and genotypes of six T cell lines established from four patients with adult T cell leukemia (ATL) were compared with those of fresh ATL cells. The surface antigen densities of CD3 and T cell receptor (TCR) complex examined by OKT3 and WT-31 mAbs on fresh ATL cells were low. But three IL-2 dependent cell lines, SKT-2, SKT-5, and ATL-17-2, established from three patients expressed a high density of CD3-TCR complex on their surfaces. On the contrary, three other cell lines (SKT-1A, SKT-1B, and MMT-2), established from two other patients, expressed a low density of CD3-TCR complex. Genotypic analysis of TCR beta chain (T beta) gene and integration sites of the proviral genome of human T cell lymphotropic virus type 1 demonstrated that SKT-1A, SKT-1B, and MMT-2 were derived from the original leukemic clones. These apparent associations between a defect of expression of CD3-TCR complex and identical genotypes of fresh and cultured cells suggest that the defect of expression of CD3-TCR complex may play a key role for development of ATL. PMID- 2972893 TI - Investigations into tracheal prosthetic reconstruction. AB - Reconstruction of circumferential tracheal defects may be necessary following trauma or oncologic surgery. For defects up to approximately 6 centimeters in an adult, an end-to-end anastomosis appears to be the method of choice. Larger defects may require utilization of a tracheal prosthesis. In dogs, a tracheal prosthesis constructed of Dacron polyurethane mesh (Osteomesh) was found to well tolerated following a two-stage implantation procedure. Normal confluent respiratory epithelium was identified in the lumen of the prosthesis. Previously reported difficulties with recurrent granulation tissue at the areas of anastomosis appear to have been successfully avoided. PMID- 2972894 TI - Changes of transcutaneous CO2 and arterial CO2 tension during laparoscopy. PMID- 2972895 TI - [70th anniversary of the People's Commissariat of Public Health of the RSFSR]. PMID- 2972896 TI - [The experience of perestroika in the work of the Bugulma Medical School]. PMID- 2972898 TI - Choice of contraceptives. PMID- 2972899 TI - T4+ (helper phenotype) chronic lymphocytic leukemia: case report with liver failure and literature review. AB - T-cell lymphomas and leukemias are a heterogeneous group of neoplasms found in each anatomic compartment of the T system (marrow, thymus, and various peripheral tissues) and that have varying phenotypic expressions. Histopathologic features of the thymic and peripheral T neoplasms do not fit into a clearly defined pattern, and clinical expressions of T neoplasms are likewise variable. This report describes a 60-year-old man with "chronic" lymphocytic leukemia of T4+ (helper) phenotype. Rapid deterioration in liver function, presumably due to tumor infiltration, was unexpected and has not previously been reported. The unusual clinical and pathologic features are reviewed in the context of T-cell neoplasms, particularly T4+ processes. PMID- 2972897 TI - New natural oestradiol/cyproterone acetate oral contraceptive for pre-menopausal women. AB - Fifty (50) healthy ovulating women aged between 35 and 47 yr (mean age 39) were randomly allocated to one of two groups treated with biphasic formulations of either oestradiol valerate/cyproterone acetate (Group A) or oestradiol valerate/norethisterone (Group B). A double-blind design was used during the first 6 mth of treatment. In Group A, 21 out of 26 women (81%), and in Group B, 16 out of 24 (67%) completed the first year of treatment. No pregnancies occurred. The mid-cycle serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) peaks were suppressed, but there were no differences between the pretreatment serum oestradiol values and those observed during the treatment cycles. The serum progesterone values indicated that only one ovulatory cycle occurred during the first year in Group A, while there were 11 in Group B. Ultrasonic studies revealed follicular growth during treatment in both groups, most follicles becoming atretic or persistent without ovulation. No significant changes were observed in 11 coagulation factors studied in 16 women in Group A. Serum total cholesterol decreased by about 10% in both groups. In Group A bleeding became scantier and dysmenorrhoea disappeared. The incidence of spotting varied between 30% and 40%, but it is important to note that the total number of bleeding days per cycle fell. The oestradiol valerate/cyproterone acetate combination was thus found to inhibit ovulation, provide tolerable cycle control and to be free from adverse metabolic side effects. PMID- 2972900 TI - [A patient with Calve-Legg-Perthes syndrome, elevated levels of creatine phosphokinase and positive screening for mucopolysaccharides]. PMID- 2972901 TI - The monoselector. An electronic aid for environmental remote control of electrical appliances for paralyzed persons. AB - A microprocessor-controlled electronic aid has been developed, which allows paralyzed persons to control up to 16 different electrical appliances using only one switch, adaptable to the nature of the handicap. This aid, the 'Monoselector', makes use of the Busch Timac X-10 remote control system. It will be produced industrially. PMID- 2972902 TI - A fuzzy model for medical diagnosis. AB - A medical diagnosis support system based on a fuzzy model for illnesses is proposed. The model for each disease is a lattice structure composed by unidirectional fuzzy relations among intermediate diagnostic units (IDU). Its evaluation is sequential: it starts at the bottom units, the observable medical evidence (symptoms), and progresses through the IDUs to the cusp unit, representing the disease. Three types of IDUs are defined: for associated, non associated and excluding evidence. These units represent medical concepts gathering knowledge under a common characteristic or criterion of diagnostic interest. Fuzzy unidirectional relations are used to quantify the medically known dependence between IDUs. The model is specified for six cardiopathies, identifying and calibrating its parameters using data from patient's records. The model performance is evaluated comparing results with the diagnosis provided by three medical observers on new patient's records. Concordance histograms are shown as an objective measure for the model performance. A discussion about application of the diagnosis sensitivity respect to symptoms and respect to IDUs relations is performed. PMID- 2972903 TI - The design of a saccade-size predictor for eye communication. AB - A method for predicting the end position of a saccade, based on the peak velocity/amplitude relationship, was studied for application in eye communication. An analogue delay line of the bucket brigade type was implemented for measuring the saccadic velocity on-line. The magnitude of the differentiated saccade-signal had been subdivided in discrete values, corresponding to the distances between the neighbouring LEDs at the characters in a row of the communication board. An up-down counter started one by one the successive switching-on of the LEDs at each step in the saccadic velocity staircase. A single emitting LED corresponded finally to a predicted eye position. The signal to-noise ratio of the detecting transistors of the infrared limbus-reflection method determined the confidence limits in prediction of the saccadic sizes. Predictions turned out to be reliable over 6 characters at 3 degrees equidistant locations in a horizontal row. It is concluded in a discussion that the feasibility of a two-dimensional fast eye keyboard system can be studied by one saccade-size predictor in combination with the magnetic field scleral search coil technique. PMID- 2972904 TI - The evaluation of glucagon infusion algorithms for a counterregulatory system in artificial endocrine pancreas. AB - As counterregulatory system of artificial endocrine pancreas, glucagon infusion algorithm has been developed and its usefulness has been examined in pancreatectomized dogs and a pancreatectomized diabetic patient. Glucagon infusion rate (G1nIR(t)) was determined depending on proportional plus derivative action to blood glucose concentration (BG(t] with the time delay (tau) to start infusion as follows. G1nIR(t) = Gp(BGp- BG(t-tau)) + Gd(-dBG(t-tau)) + Gc where BGp is the projected value of blood glucose concentration, Gp, and Gd are the coefficients and Gc is the constant for basal glucagon supplement. Glucagon infusions based only on proportional action (Gp/Gd/Gc/tau) = (0.2/0/0.4/10 or 0.4/0/0.4/10) failed in simulating the pattern of blood glucose or glucagon response seen in normal dogs. Glucagon infusion based on proportional plus derivative action (Gp/Gd/Gc/tau = 0.2/0.4/0.4/10) successfully mimicked the pattern of blood glucose concentration and plasma glucagon profile seen in normal dogs. This glucagon infusion algorithm has been applied to control insulin induced hypoglycemia in a pancreatectomized patient with the same infusion parameters. Hypoglycemia was recovered to normoglycemia in 80 min and the plasma glucagon patterns showed no significant difference from those in healthy volunteers. These data indicate that glucagon infusion algorithm thus developed is effective to render hypoglycemia to normoglycemia and mimic plasma glucagon response seen in normal subjects. PMID- 2972905 TI - Expert-system-based fuzzy control of arterial pressure by drug infusion. AB - This paper proposes a new type of fuzzy controller which is based on the general purpose Fuzzy LOgic Production System shell FLOPS [1, 2, 7, 12, 13]. Human mean arterial pressure was controlled by the proposed fuzzy controller, in digital computer stimulation, by regulating the infusion rate of the drug, sodium nitroprusside. Under nonstationary arterial background pressure noise, mean arterial pressures of different patients with different sensitivities to sodium nitroprusside were controlled satisfactorily. Furthermore, the effects of changing the adjustable parameters of the proposed fuzzy controller on the control results were studied. Finally, the performance of the proposed fuzzy controller was compared to that of a fine-tuned Proportional-Integral (PI) controller by using a first-order linear model. PMID- 2972906 TI - [Intratumoral administration of OK432 in rats with liver tumor]. AB - The approach to the treatment of unresectable liver tumor involves immunotherapy. Systemic administration of OK432 has been widely used in the treatment of malignant neoplasms. However, the most potent antitumor activity of the drug may be expected when it is administered intratumorally. The author evaluated the effect of intratumoral injection of OK-432 on the survival time, the immunological parameters (such as the NK activities of spleen cells and peritoneal exudate cells and the interferon production by spleen cells) and the tumor infiltrating lymphocytes (TIL) in the rats with liver tumor induced by feeding the hepatocarcinogen, 3'-methyl-4-di-methyl-aminoazobenzene. The mean survival time was significantly longer in the rats injected with OK432 intratumorally (I.T. group) than in the rats injected with OK432 intraperitoneally only (I.P. group) and in the rats injected with normal saline intratumorally (Control group). The immunological parameters significantly improved in the rats of I.T. group than in the controls. Intratumoral injection of OK432 increased the number of TIL, especially NK cells and suppressor/cytotoxic T cells. These beneficial effects could be responsible for the better survival time in the rats of I.T. group. The author concluded that the intratumoral OK432 administration therapy is effective for the treatment of the patients with unresectable liver tumor. PMID- 2972908 TI - Rad3 protein of Saccharomyces cerevisiae: overexpression and preliminary characterization using specific antibodies. AB - The cloned RAD3 gene of Saccharomyces cerevisiae was tailored into expression vectors for overexpression of Rad3 protein in Escherichia coli and in yeast. In both organisms the overexpressed protein is detected as a species of molecular weight ca. 90 kDa, the size expected from the sequence of the cloned gene. The protein overexpressed in E. coli is largely insoluble; however the insoluble fraction was used to generate affinity-purified polyclonal antisera which proved to be powerful reagents for the initial characterization of Rad3 protein expressed in yeast. These studies showed that: (1) when overexpressed in yeast most of the Rad3 protein is detected in the soluble fraction of cell extracts; (2) endogenous Rad3 protein is untransformed cells is also ca. 90 kDa in size and is located in the cell nucleus; (3) Rad3/beta-galactosidase fusion protein partially purified on an affinity matrix is associated with DNA-dependent ATPase activity that is inhibited in the presence of anti-Rad3 antibodies, suggesting that Rad3 protein is an ATPase; and (4) Rad3 antibodies cross-react with two electrophoretically distinguishable polypeptides present in the nuclear fraction of human cells, and with a single polypeptide in extracts of Drosophila cells. PMID- 2972907 TI - In vivo effect of DNA repair on the transition frequency produced from a single O6-methyl- or O6-n-butyl-guanine in a T:G base pair. AB - We have previously reported some effects of DNA repair on the transition frequencies produced by an O6-methyl-guanine (MeG) or an O6-n-butyl-guanine (BuG) paired with C at the first position of the third codon in gene G of bacteriophage phi X174 form I' DNA (Chambers et al. 1985). We now report experiments in which the transition is produced from T:MeG or T:BuG, instead of C:MeG or C:BuG, located at this site. The site-modified DNAs were transfected into cells with normal DNA repair as well as into cells with repair defects (uvrA, uvrB, uvrC, recA, uvrArecA). The lysates were screened for phage carrying the expected transition using a characteristic change in phenotype. The data demonstrate that the transition frequency from T:BuG is low (0.3% of total phage progeny) in cells with normal repair (Escherichia coli AB1157) and increases 7-fold in uvrA cells (E. coli AB1886). A similar increase is seen in uvrB and uvrC cells (AB1885, AB1884). These data, like our previous data, indicate BuG is repaired primarily by excision. In contrast to this, the transition frequency from T:MeG is high (5 +/- 2%) in cells with normal repair. After induction of alkyl transfer repair in E. coli AB1157, the transition frequency goes up 5-fold. Compared with cells with normal repair, the transition frequency goes up 2-fold in uvrA, uvrB and uvrC cells; it goes up 1.5-fold in recA cells (E. coli AB2463). The data reinforce our earlier conclusion that MeG is repaired primarily by alkyl transfer, but the ABC excinuclease as well as RecA protein inhibit this repair process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972910 TI - Increased expression of the Escherichia coli umuDC operon restores SOS mutagenesis in lexA41 cells. AB - The lexA41 allele of Escherichia coli encodes a semidefective mutant repressor that is also resistant to RecA facilitated cleavage. Cells harboring the lexA41 allele were found previously to repress only a subset of operons in the SOS regulon. lexA41 cells cannot promote SOS mutagenesis, presumably because one or more operons required for mutagenesis are repressed by this mutant repressor. Using the lac regulatory system to increase the expression of the umuDC operon, we were able to restore mutagenesis in the lexA41 mutant. We conclude that the products of the umuDC operon appear to be uniquely limiting in this mutant. PMID- 2972909 TI - Nature of the SOS mutator activity: genetic characterization of untargeted mutagenesis in Escherichia coli. AB - In Escherichia coli, induction of the SOS functions by UV irradiation or by mutation in the recA gene promotes an SOS mutator activity which generates mutations in undamaged DNA. Activation of RecA protein by the recA730 mutation increases the level of spontaneous mutation in the bacterial DNA. The number of recA730-induced mutations is greatly increased in mismatch repair deficient strains in which replication errors are not corrected. This suggests that the majority of recA730-induced mutations (90%) arise through correctable, i.e. non targeted, replication errors. This recA730 mutator effect is suppressed by a mutation in the umuC gene. We also found that dam recA730 double mutants are unstable, segregating clones that have lost the dam or the recA mutations or that have acquired a new mutation, probably in one of the genes involved in mismatch repair. We suggest that the genetic instability of the dam recA730 mutants is provoked by the high level of replication errors induced by the recA730 mutation, generating killing by coincident mismatch repair on the two unmethylated DNA strands. The recA730 mutation increases spontaneous mutagenesis of phage lambda poorly. UV irradiation of recA730 host bacteria increases phage untargeted mutagenesis to the level observed in UV-irradiated recA+ strains. This UV-induced mutator effect in recA730 mutants is not suppressed by a umuC mutation. Therefore UV and the recA730 mutation seem to induce different SOS mutator activities, both generating untargeted mutations. PMID- 2972912 TI - Regulation of cardiac sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase. AB - The two high affinity calcium binding sites of the cardiac (Ca2+ + Mg2+)-ATPase have been identified with the use of Eu3+. Eu3+ competes for the two high affinity calcium sites on the enzyme. With the use of laser-pulsed fluorescent spectroscopy, the environment of the two sites appear to be heterogeneous and contain different numbers of H2O molecules coordinated to the ion. The ion appears to be occluded even further in the presence of ATP. Using non-radiative energy transfer studies, we were able to estimate the distance between the two Ca2+ sites to be between 9.4 to 10.2 A in the presence of ATP. Finally, from the assumption that the calcium site must contain four carboxylic side chains to provide the 6-8 ligands needed to coordinate calcium, and based on our recently published data, we predict the peptidic backbone of the two sites. PMID- 2972911 TI - Allosteric regulation of cardiac sarcoplasmic reticulum Ca-ATPase: a comparative study. AB - The mechanisms of allosteric regulation of the Ca-ATPases of cardiac and skeletal sarcoplasmic reticulum by ATP have been compared. Although both enzymes showed stimulation of ATPase activity by ATP, the cardiac enzyme did not show the plateau in ATPase activity at 10-100 microM ATP seen with the skeletal enzyme. Likewise the phosphoenzyme (EP) levels did not plateau with the cardiac enzyme as they did with the skeletal enzyme. The apparent negative cooperatively which was seen in the kinetics of ATP hydrolysis at low ATP concentrations was not due to negative cooperatively in substrate binding to either enzyme. The cardiac enzyme did show, however, much higher affinity for the ATP analog, AMPPCP, which helps explain how AMPPCP blocks ATPase activity in the cardiac enzyme and stimulates ATPase activity in the skeletal enzyme. Fluorescein isothiocyanate was used to determine if allosteric regulation takes place through site-site interactions in oligomers. The 1 to 1 ratio between AMPPCP binding sites and FITC binding sites eliminated allosteric regulation by effector sites in both enzymes. The allosteric mechanism which remained was one in which the active-site becomes an effector-site by the early departure of ADP in the reaction mechanism. The step stimulated by the binding of ATP at the active-site turned effector-site was a nonphosphorylated form of the enzyme in cardiac sarcoplasmic reticulum and a phosphorylated form in skeletal sarcoplasmic reticulum. PMID- 2972913 TI - Molecular topography of the Ca2+-ATPase of sarcoplasmic reticulum. AB - The Ca2+ binding site region of the Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum was labeled with several fluorescent analogs of dicyclohexylcarbodiimide. As has been shown by Chadwick and Thomas, in the absence of Ca2+ in the medium, labeling with the naphthyl carbodiimide results in the inhibition of enzyme activity. Further, Ca2+ occupancy of the high affinity sites of the enzyme protects against incorporation into the site(s). The fluorescent carbodiimide has been used to determine the depth of the site of label incorporation relative to the aqueous-bilayer interfaces by quenching studies using spin-labeled fatty acid derivatives. The series of quenchers used have their spin-label moiety located at different positions along the fatty acid chain. It was found that after suitable correction for differences in partitioning of the various derivatives, the order of quenching efficiency was 16 greater than 12- greater than 10- greater than 7- greater than 5-NS, indicating that the naphthyl moiety is near the center of the bilayer. In contrast, quenching with the aqueous-restricted I- indicated that the label is accessible from the external milieu, likewise for a presumed aqueous quencher, acrylamide. The aqueous quenchers accessibilities were altered upon Ca2+ binding to the ATPase. Quenching of the intrinsic fluorescence with the x-NS derivatives indicates that the ATPase tryptophan residues are primarily localized at the aqueous-membrane interfaces, with the order of quenching being 5- greater than 7- greater than 10- greater than 12- greater than 16-NS. The trp residue(s) which changes its fluorescence upon Ca2+ binding is shown to be near the membrane surface. PMID- 2972915 TI - Sarcolemmal Na+-Ca2+ exchange and Ca2+-pump activities in cardiomyopathies due to intracellular Ca2+-overload. AB - In order to identify defects in Na+-Ca2+ exchange and Ca2+-pump systems in cardiomyopathic hearts, the activities of sarcolemmal Na+-dependent Ca2+ uptake, Na+-induced Ca2+ release, ATP-dependent Ca2+ uptake and Ca2+-stimulated ATPase were examined by employing cardiomyopathic hamsters (UM-X7.1) and catecholamine induced cardiomyopathy produced by injecting isoproterenol into rats. The rates of Na+-dependent Ca2+ uptake, ATP-dependent Ca2+ uptake and Ca2+-stimulated ATPase activities of sarcolemmal vesicles from genetically-linked cardiomyopathic as well as catecholamine-induced cardiomyopathic hearts were decreased without any changes in Na+-induced Ca2+-release. Similar results were obtained in Ca2+ paradox when isolated rat hearts were perfused for 5 min with a medium containing 1.25 mM Ca2+ following a 5 min perfusion with Ca2+-free medium. Although a 2 min reperfusion of the Ca2+-free perfused hearts depressed sarcolemmal Ca2+-pump activities without any changes in Na+-induced Ca2+-release, Na+-dependent Ca2+ uptake was increased. These results indicate that alterations in the sarcolemmal Ca2+-efflux mechanisms may play an important role in cardiomyopathies associated with the development of intracellular Ca2+ overload. PMID- 2972914 TI - Estimation of inter-binding-site distances in sarcoplasmic reticulum (Ca2+ + Mg2+)-ATPase under occluded and non-occluded conditions. AB - Distances between binding sites in skeletal sarcoplasmic reticulum (Ca2+ + Mg2+) ATPase were estimated by measuring energy transfer between a luminescent lanthanide probe for the calcium sites (Eu3+) and suitable acceptors. A distance measurement between the two calcium binding sites of the enzyme was made with and without MgATP, corresponding to two different states in the hydrolytic cycle. We found that without ATP, the inter-calcium-site distance is about 0.9 nm, while in the presence of MgATP these sites are 0.05 nm to 0.1 nm closer. We also estimated the distance from the calcium sites to the hydrolytic site using CrATP as an acceptor; this distance is approximately 1.8 nm. These measurements provide the start for a description of intramolecular movement during the transport cycle, and should be of use in mapping out the three-dimensional structure of the Ca2+ pump. PMID- 2972916 TI - [Kinetic model of the action of oligomeric enzymes using phosphofructokinase as an example. II. General formulation of the hierarchical model]. AB - A general approach is suggested to describe the steady-state kinetics of the oligomeric enzymes on the base of the generalized statistical Ising model. Detailed analysis is given for the case of a oligomeric enzyme with a hierarchical supramolecular organization. A protomer of this enzyme composed of several equivalent subunits represents the quarternary level of structure. In their turn the finite or infinite number of protomers is associated into a oligomer thus creating a new "quinternary" level of the enzyme organization. The model accounts for the ligand-induced homotrophic cooperative interactions: firstly, between the neighbouring protomers and secondly, between the subunits of the same protomer. The influence of protomer conformation on the subunit state and the cooperativity induction caused by two-ligand binding are also taken into consideration. Monod-Wyman-Changeux's and Koshland's models are shown to be special limit cases of the suggested general theory. PMID- 2972917 TI - Monomeric and dimeric IgG1 as probes for assessing high-affinity and low-affinity receptors for IgG on human monocyte-derived macrophages and on activated macrophages. AB - From a panel of IgG1 myeloma proteins, only one was found to interact with human monocyte FcR in a manner similar to that of polyclonal IgG. This protein was used in binding studies involving human macrophage Fc receptors. A monomeric fraction depleted of dimeric and polymeric IgG1 was crosslinked with bis-diazonium benzidine, and a fraction highly enriched in cross linked IgG1 dimers was radiolabeled. Labeled monomeric and dimeric IgG were allowed to interact with monocytes that had matured to macrophages in vitro. The association with macrophages at 4 degrees C, in the presence of cytochalasin B, reached a plateau after 6 hr. The dissociation induced by excess unlabeled IgG followed similar kinetics as the association, but 20-30% of the bound IgG could not be dissociated. Under equilibrium conditions, evidence for a single FcR population binding monomeric IgG was obtained, the Kd being in the range of 12-42 nM. In contrast, the binding of dimeric IgG was more consistent with a model assuming two populations of binding sites when appropriate curve-fitting calculations were applied. The high-affinity FcR population had a Kd in the range of 0.8-3.5 nM, whereas the Kd of the low-affinity FcR population was in the range of 28-85 nM. When macrophages had been pre-treated with recombinant interferon-gamma, the expression of high-affinity sites was increased by a factor of 1.5-3, but the number of low-affinity sites was not augmented. Cytofluorographic analyses confirmed the increased expression of high-affinity FcR, binding fluoresceinating murine IgG2a. The expression of CD16, a low-affinity FcR expressed on neutrophils, NK cells and macrophages, as well as the expression of the complement receptor type III was little influenced by the rIFN-gamma pretreatment. PMID- 2972919 TI - The Haversian canal network in the femoral compact bone in some vertebrates. PMID- 2972918 TI - On the 65th anniversary of the birth of Professor Dr. Tiberiu Maros. PMID- 2972921 TI - Isolation and cultivation of rabbit endocardial endothelial cells. Preliminary data. PMID- 2972920 TI - Cell-to-cell herniae in the media of coronary arterioles. PMID- 2972922 TI - Morphological criteria of diagnosis in lung tissue biopsies from diffuse interstitial pulmonary diseases. PMID- 2972923 TI - Carcinoma of the esophagogastric junction. PMID- 2972924 TI - Morphology of a plurihormonal human pituitary adenoma composed of FSH, LH and TSH cells. PMID- 2972925 TI - [Victor Babes and his works on congenital anomalies]. PMID- 2972926 TI - Radiosensitive Down syndrome lymphoblastoid lines have normal ionizing-radiation induced inhibition of DNA synthesis. AB - The extent of X-ray-induced inhibition of DNA synthesis was determined in radiosensitive lymphoblastoid lines from 3 patients with Down syndrome and 3 patients with ataxia telangiectasia (AT). Compared to 6 normal control lines, the 3 AT lines were abnormally resistant to X-ray-induced inhibition of DNA synthesis, while the 3 Down syndrome lines had normal inhibition. These results demonstrate that radiosensitive human cells can have normal X-ray-induced inhibition of DNA synthesis and provide new evidence for the dissociation of radiosensitivity from radioresistant DNA synthesis. PMID- 2972927 TI - Isomyosin changes after functional electrostimulation of denervated sheep muscle. AB - Isomyosin analyses by biochemical, immunochemical, and histochemical investigations have been carried out in five sheep following unilateral recurrent laryngeal nerve paralysis and direct functional electrostimulation of the denervated cricoarytenoid posterior muscle. Myosin light chains were identified by two-dimensional gel electrophoresis. Myosin heavy chains were analyzed by one dimensional SDS-polyacrylamide gel electrophoresis. Slow myosin heavy chain was identified by orthogonal peptide mapping and immunochemistry. The stimulation effect at cellular level was determined using adenosine triphosphatase (ATPase) histochemistry. A dramatic increase of the type 1 fiber area (slow, fatigue resistant fibers) could be seen after many weeks of an increasing regime of low frequency direct electrical stimulation. Biochemically, the amount of slow myosin was always higher than in normal muscles. Some muscles were transformed almost completely to the slow type. At the time they were studied and with the methods employed, the expression of embryonic isomyosin was not observed. In conclusion, after numerous weeks of maintained functional activity, elicited by direct electrostimulation, the denervated muscle regionally showed areas of hypertrophy or at least lack of atrophy of slow myofibers without major signs of muscle damage. PMID- 2972928 TI - Identification and characterization of cDNA clones encoding antigens of Eimeria tenella. AB - An Eimeria tenella cDNA library was constructed in the expression vector lambda gt11 from poly (A+) RNA extracted from sporulating oocysts. The library was screened with rabbit antiserum raised against antigens extracted from fully sporulated oocysts. All of the antigen-expressing plaque-purified clones were initially characterized by cross screening with antisera raised against different stages of the E. tenella life cycle, as well as with antiserum raised against sporozoites of a related species, namely E. acervulina. A selected number of clones were further characterized by antibody selection coupled with immunoblotting and DNA cross hybridization. Three different E. tenella antigens were identified. All three appear to be constitutively expressed at the protein level during sporogony. PMID- 2972929 TI - Specific serodiagnosis of visceral leishmaniasis using a Leishmania donovani antigen identified by expression cloning. AB - A lambda gt11 expression library was constructed using mRNA from the promastigote form of Leishmania donovani (African isolate MHOM/ET/67/HU3). The library was screened with serum obtained from a patient who contracted visceral leishmaniasis in the Sudan. Several cDNA clones which expressed beta-galactosidase/L. donovani antigen fusion proteins were isolated. One of these clones corresponded to a 60 kDa membrane-associated antigen. By a Western blot assay antibodies against the fusion protein were detected universally in the sera of visceral leishmaniasis patients. They were not detected in sera from patients with cutaneous leishmaniasis or other parasitic protozoan infections. The gene coding for this antigen was specific to the genus Leishmania as judged by DNA hybridisation, and its chromosomal location was conserved. A 20 kb mRNA was detected on Northern blots of promastigote RNA. PMID- 2972930 TI - Purification, characterization, and immunochemical studies of beta-N-acetyl-D hexosaminidase from the parasitic nematode Trichinella spiralis. AB - The exoglycosidase, beta-N-acetyl-D-hexosaminidase was purified 600-fold from the muscle-stage larvae (L1) of Trichinella spiralis. On sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the purified enzyme-active fraction contained 4 polypeptides with apparent molecular weights of 100,000, 68,000, 58,000 and 54,000. The beta-N-acetyl-D-hexosaminidase corresponds to the Mr 100,000 polypeptide as demonstrated by SDS-PAGE analysis of the enzyme-stained region isolated from a non-denaturing polyacrylamide gel. In addition, rabbit antiserum to a homogeneous preparation of the Mr 100,000 polypeptide (isolated by electroelution from an SDS-PAGE gel) specifically immunoprecipitated beta-N acetyl-D-hexosaminidase activity from an extract of L1. Isoelectrofocusing (pH 3 10) resolved 4 isoenzymes of T. spiralis beta-N-acetyl-D-hexosaminidase with isoelectric points (pI) of 5.35, 5.49, 5.63 and 5.79. The T. spiralis beta-N acetyl-D-hexosaminidase is a glycoprotein based on its binding to lentil-lectin Sepharose affinity column and its specific binding of concanavalin A on Western blots. The IgG fraction of T. spiralis-infected mouse serum specifically immunoprecipitated T. spiralis beta-N-acetyl-D-hexosaminidase. The removal of carbohydrate from T. spiralis beta-N-acetyl-D-hexosaminidase significantly reduced its antigenicity. Immunocytochemical analysis of L1 tissue sections with polyclonal rabbit antisera to the homogeneous beta-N-acetyl-D-hexosaminidase enzyme indicated localization on cell membranes and the epicuticle. PMID- 2972931 TI - Purification of Plasmodium falciparum digestive vacuoles and partial characterization of the vacuolar membrane ATPase. AB - Plasmodium falciparum digestive vacuoles containing ferric oxide granules were purified from parasite homogenates by centrifugation on discontinuous sucrose gradients. Digestive vacuole membranes prepared by osmotic lysis and washed with KCl showed no detectable contamination by erythrocyte membrane proteins and only minimal contamination by non-vacuolar parasite proteins. Purified vacuolar membranes were 2.6-fold enriched in total parasite membrane ATPase activity. This ATPase was optimally active at pH 7 in the presence of at least 2 mM Mg2+. Ca2+ and Mn2+ were approximately 80-90% as effective as Mg2+, and Zn2+, Co2+ and Fe2+ also exerted some stimulatory effect. The vacuolar membrane also hydrolyzed GTP, UTP, CTP and ADP, but AMP and 3',5'-cyclic AMP were hydrolyzed only one-tenth as effectively as ATP. The ATPase was unaffected by vanadate, ouabain or oligomycin but was significantly inhibited by the proton pump inhibitors NEM and NBD-Cl. Of 6 antimalarial drugs tested, quinine and quinacrine were the most effective inhibitors and mefloquine was the least effective. PMID- 2972932 TI - Microtubule motors. Cytoplasmic dynein graduates. PMID- 2972933 TI - Abnormal differentiation of thymocytes in mice treated with cyclosporin A. AB - Cyclosporin A (CsA) acts as a powerful immunosuppressive agent, and also, when given in repeated doses, can cause T-cell-dependent graft-versus-host disease and organ-specific autoimmune disease in rodents. This suggests that CsA interferes with the processes governing self-tolerance, either by nullifying the activity of T suppressor cells or by preventing the deletion of autoreactive T cells during ontogeny in the thymus. We report here that irradiated mice given repeated injections of CsA show striking dysfunction of the thymus. There are two different effects, the first of which is that CsA seems to block the differentiation of immature CD4+CD8+ thymocytes into mature CD4+CD8- and CD4-CD8+ cells expressing a high density of T-cell receptors and CD3 molecules. Second, CsA-treated mice show incomplete deletion of T cells expressing T-cell receptor molecules reactive to self H-2 I-E molecules. PMID- 2972935 TI - Pain management programs. Clinical and research perspectives. PMID- 2972934 TI - Effect of metergoline, fenfluramine, and 8-OHDPAT on catalepsy induced by haloperidol or morphine. AB - The influences of the indirect serotonin agonist fenfluramine (5; 10 mg/kg s.c.), the serotonin antagonist metergoline (5; 10 mg/kg s.c.) and the 5-HT1A agonist 8 OHDPAT (0.1; 0.2; 0.46 mg/kg s.c.) on haloperidol-induced catalepsy in rats or mice and on morphine-induced catalepsy in rats were studied. Morphine-induced catalepsy was enhanced by fenfluramine and attenuated by metergoline, whereas neither fenfluramine nor metergoline had any effect on haloperidol-induced catalepsy. 8-OHDPAT strongly antagonised catalepsy induced by morphine or haloperidol. We conclude that serotonergic transmission plays a major role in effectuating morphine catalepsy but not in effectuating haloperidol catalepsy. The antagonistic effect of 8-OHDPAT suggests a secondary, modulating role for 5 HT1A receptor mediated events in both types of catalepsy. PMID- 2972936 TI - [Medicine, servant of mercy]. PMID- 2972938 TI - Effects of serotonin receptor antagonists and agonists on the tail-flick response in mice involve altered tail-skin temperature. AB - Tail-flick latency and tail-skin temperature were measured in mice after administration of serotonin (5-HT) receptor antagonists (metergoline and metitepin) and agonists [5-methoxy-N,N-dimethyltryptamine (5-MeODMT) and 8 hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT)]. Metergoline (4 mg/kg) and metitepin (0.5 mg/kg) reduced the tail-flick latencies and increased the tail skin temperatures, but the effect on the tail-flick latencies was non-significant when the effect of temperature was taken into account. Both 5-MeODMT (3 mg/kg) and 8-OH-DPAT (1 mg/kg) reduced the tail-skin temperature but only 5-MeODMT increased the tail-flick latencies. The effect of 5-MeODMT on tail-flick latencies was still highly significant when the effect of temperature was taken into account. The results show that the apparent hyperalgesia elicited by 5-HT receptor antagonists in the tail-flick test in the mouse is secondary to increased tail-skin temperature and not due to increased nociceptive sensitivity. The antinociceptive effect of 5-MeODMT in the tail-flick test can, however, not be explained by effects of temperature. PMID- 2972937 TI - Differential effects of pituitary stalk-section on posterior pituitary and hypothalamic contents of prolactin-releasing factor, oxytocin, dopamine and beta endorphin. AB - We have recently shown that the posterior pituitary (neurointermediate lobe) contains a potent prolactin (PRL)-releasing factor (PRF) which is distinct from known PRL secretagogues. Posterior pituitary PRF appears to be a small peptide of an unknown cellular origin. Using pituitary stalk-sectioned (SS) male rats, the objectives of this study were: (1) to determine if PRF is transported from the hypothalamus or is synthesized within the pituitary gland, and (2) to compare changes in PRF activity with alterations in the posterior pituitary content of beta-endorphin (beta-END), oxytocin (OXY), and dopamine (DA). One or two weeks following pituitary SS or sham surgery (SHAM), acid extracts of the posterior pituitary and medial basal hypothalamus (MBH) were analyzed for their hormone content. PRF activity was assessed by determining the stimulation of PRL secretion from perifused anterior pituitary cells, DA was measured by HPLC, and OXY and beta-END levels were determined by RIAs. OXY and DA concentrations in the posterior pituitary were reduced more than 95% at both 1 and 2 weeks after SS. PRF activity in the posterior pituitary was significantly reduced by 75 and 90%, 1 and 2 weeks after SS, respectively. In contrast, beta-END levels in the posterior pituitary at these times increased 20 and 60%, as compared to SHAM rats. Unlike the posterior pituitary, OXY levels in the MBH increased 123% 1 week following SS, and 1,260% at 2 weeks. These increases may reflect the accumulation of OXY-containing secretory vesicles in the severed nerves. DA concentrations in the MBH showed a biphasic pattern. DA levels were initially decreased by 70%, and then increased, but remained 30% below SHAM levels. The reason for these alterations in DA levels is not clear.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2972941 TI - Midline lumbar ganglion/synovial cyst mimicking an epidural tumor: case report and review of pathogenesis. AB - A case of a midline lumbar extradural ganglion/synovial cyst causing lumbar canal stenosis and mimicking an epidural tumor is presented. The lesion was demonstrated by a magnetic resonance imaging study, and relief of symptoms was achieved with decompressive laminectomy and total removal of the mass. The pathogenesis of lumbar ganglion/synovial cyst is reviewed. PMID- 2972940 TI - Microtrauma in the lumbar spine: a cause of low back pain. AB - Excessive mechanical stress on the intervertebral disc may be one of the causes of low back pain. Most studies testing this thesis, however, have been based on quantification of the mechanical response of functional units at failure. Typically, radiography is used to demonstrate trauma to the vertebral body at the failure load. The description of failure and radiographic demonstration of damage are meaningful in specifying the tolerance limits of the structure. It is important, however, to understand the sequence underlying the initiation of injury, which may occur at subfailure physiological loads. In this study, we identified the initiation of injury to the lumbar spine by subjecting functional units to axial compressive loads using the mechanical response as a basis. Because conventional radiography failed to detect trauma at this level, advanced sectioning techniques were used. The initiation of injury (microtrauma) is defined as the point on the load-deflection curve where the structure exhibits a decreasing level of resistance for the first time before reaching its ultimate load-carrying capacity. The load deflection curve on this basis was classified into the ambient or preload phase, physiological loading phase, traumatic phase, and post-traumatic phase. Structures loaded to the end of the physiological loading phase did not exhibit any yielding or microtrauma. Injury in the form of microfractures of the endplate not detected on radiography, however, was observed under cryomicrotomy for structures loaded into the traumatic loading phase. PMID- 2972939 TI - Beta-endorphin-(1-27) antagonizes beta-endorphin- but not morphine-, D-Pen2-D Pen5-enkephalin- and U50, 488H-induced analgesia in mice. AB - beta-Endorphin-(1-27), administered intraventricularly has been previously reported to block the analgesia induced by beta-endorphin injected intraventricularly. The present study was to determine if the blocking effect of beta-endorphin-(1-27) was specific to beta-endorphin which stimulates epsilon receptors, but not to other opioids with activity at different opioid receptors. The antagonistic effects of beta-endorphin-(1-27) on the analgesia induced by beta-endorphin (epsilon-opioid receptor agonist), D-Ala2-NMePhe4-Gly-ol enkephalin(DAGO) and morphine, (mu-opioid receptor agonists), D-Pen2-D-Pen5 enkephalin(DPDPE) and D-Ala2-D-Leu5-enkephalin(DADLE) (delta-opioid receptor agonists) and U-50, 488H (kappa-opioid receptor agonist) were studied. beta Endorphin-(1-27) injected intraventricularly, at doses which, when injected alone did not produce analgesia, antagonized the analgesia induced by beta-endorphin given intraventricularly. However, the analgesia induced by DAGO, morphine, DPDPE, DADLE and U-50, 488H given intraventricularly was not antagonized by beta endorphin-(1-27). The data suggest that beta-endorphin-(1-27) selectively blocks the analgesia induced by the stimulation of epsilon receptors but not by the stimulation of mu, delta, and kappa receptors. The results support the previously proposed hypothesis that beta-endorphin produces its analgesia by stimulating specific epsilon receptors. PMID- 2972942 TI - Autoradiographic localization of D1 and D2 dopamine receptors in the human brain. AB - The distribution of dopamine D1 and D2 receptors in several human brain regions was investigated using autoradiography with the radioligands [3H]SCH 23390 and [3H]spiroperidol. The highest densities of both dopamine receptor types are seen in the nucleus caudatus, putamen and nucleus accumbens. Whereas the density of the D2 receptors is similar in the two segments of the globus pallidus, the pars medialis of the globus pallidus contains a three-fold higher concentration of D1 receptors than the pars lateralis. D1 and D2 receptors are present in the amygdala and substantia nigra. Both receptor types are absent in the cerebellum. The thalamus contains low densities of D1 receptors but no D2 receptors. Only D2 receptors are seen in the anterior lobe of the pituitary gland. The whole cerebral cortex is rich in D1 receptors, while D2 receptors, in low concentrations, are confined to the entorhinal area and cingulate cortex. PMID- 2972943 TI - Presence of T-cytotoxic suppressor and leucocyte common antigen positive cells in Alzheimer's disease brain tissue. AB - Cells expressing leucocyte common antigen (LCA) as well as T-cytotoxic-suppressor (T8) and T-helper-inducer (T4) antigens were detected in significant numbers in the hippocampus and temporal cortex of Alzheimer but not normal brain tissue. Leucocytes and reactive microglia strongly expressed LCA, as well as the class II major histocompatibility complex (MCH) glycoprotein HLA-DR, but astrocytes stained negatively. Such staining indicates the immunocompetent status of microglia, as well as their phenotypic linkage to blood borne mesodermal lines. T8 and T4 positive lymphocytes were detected in capillaries and the surrounding matrix in smaller numbers than LCA positive cells. The MHC class I (HLA-A,B,C) antigens were detected on capillaries in both Alzheimer and control tissue. The significance of LCA, T4 and T8 cells in Alzheimer's disease is unknown but may be indicative of a cell mediated immune response in the disease. PMID- 2972945 TI - Industrial deafness. PMID- 2972944 TI - Groups with special health care needs. AB - Inadequate access to health services is a major problem for disadvantaged groups and the reasons for this presents health researchers with a special challenge. These groups include women, Maori people, the economically deprived, the handicapped and the geographically isolated. Maori groups are now advocating community and Marae-based health clinics whilst well women clinics have evolved from the women's health lobby. These initiatives have allowed the respective groups to take responsibility for their own health and be involved in the health decision-making process. With respect to the chronically disabled there is a continuing need for realistic support and careful evaluation. PMID- 2972947 TI - A state Supreme Court ruling in an AIDS case. PMID- 2972946 TI - The dose of hepatitis B vaccine. PMID- 2972948 TI - A one visit procedure to stabilize mobile anterior mandibular teeth. Case report. PMID- 2972949 TI - A laparoscopic approach can be applied to most cases of ectopic pregnancy. AB - Recent studies have demonstrated lower cost and morbidity associated with laparoscopic treatment of ectopic pregnancy. However, the applicability of these techniques to unselected cases of ectopic pregnancy has not yet been proved. To test the efficiency of the laparoscopic approach, an attempt was made to apply these techniques to all patients who presented with ectopic pregnancy at a medical center during a 6-month period and who met the entry criteria of hemodynamic stability; location other than abdominal with fetus, interstitial, or cervical; and visibility of the mesosalpinx. None of the 22 patients referred to the study had to be excluded because of these criteria, indicating the relative infrequency of these contraindications. All were treated successfully by laparoscopic salpingectomy (N = 12), salpingostomy (N = 8), or removal of implantations from peritoneal surfaces (N = 2). The average size of the ectopic gestation was 3.9 cm, the average amount of hemoperitoneum was 168 mL, the average additional blood loss attributable to the procedure was 30 mL, and the average drop in hematocrit postoperatively was 4.7%. The average hospital stay was 1.1 days, and patients were released for work in 1-2 weeks. An analysis of the three patients not referred to the study indicated that they may have been appropriate candidates for laparoscopic resection. This study indicates that the majority of unselected patients with ectopic pregnancy may be treated by laparoscopic procedures, with low cost and low morbidity. PMID- 2972950 TI - [Proceedings of the 38th scientific congresses of the Federation of Public Health Physicians. 15-19 May 1987, Mainz]. PMID- 2972951 TI - [In honor of the administrative councilor a.D. Dr. med. Wolfgang Nusche on the award of the Johann-Peter-Frank medal]. PMID- 2972952 TI - [Historical aspects of the control of so-called communicable diseases]. PMID- 2972953 TI - [Biotechnologic aspects and environmental health significance of in situ restoration measures of soil and ground water]. PMID- 2972955 TI - [Behavior of some pathogenic and other microorganisms in ground water with reference to delineation of water protection zones]. PMID- 2972954 TI - [Preventive health protection--assessment of legal possibilities and limits in agriculture]. PMID- 2972957 TI - [Treatment of water from flowing streams]. PMID- 2972956 TI - [Anthropogenic pollutants in drinking water--preventive responsibilities of public health offices]. PMID- 2972958 TI - [The public health significance of drinking water constituents]. PMID- 2972959 TI - [AIDS in Rhineland Pfalz--an interim evaluation]. PMID- 2972960 TI - [Legal stipulations of the AIDS disease in West Germany 1988]. PMID- 2972962 TI - [Future issues for pediatricians]. PMID- 2972961 TI - [The toxicity of fluorides]. PMID- 2972963 TI - [Neuromuscular diseases in childhood and adolescence]. PMID- 2972964 TI - [Clinical picture, diagnosis and treatment possibilities of minimal brain dysfunction]. PMID- 2972965 TI - [Tuberculin intradermal test--comparative studies]. PMID- 2972966 TI - [Psychosocial aspects of managing diabetic children and adolescents]. PMID- 2972967 TI - [Tuberculosis in West Germany--prevention, control and monitoring by the public health service]. PMID- 2972968 TI - [Allergenic, chemically irritating or toxic causes of obstructive respiratory tract disease]. PMID- 2972969 TI - [The development of mass chest x-ray units in West Germany--report of their status and future perspectives]. PMID- 2972970 TI - [Social psychiatric management of chronic psychiatric patients]. PMID- 2972971 TI - [Liberation from the fortress--on the gerontopsychiatric work of the social psychiatric service]. PMID- 2972973 TI - [Drug substitution with methadone--pro arguments]. PMID- 2972972 TI - [Alcohol in training and occupation--possible preventive management strategies by the public health service]. PMID- 2972974 TI - [Drug substitution with methadone--contra arguments]. PMID- 2972975 TI - [Responsibilities and perspectives of social psychiatric services of public health offices]. PMID- 2972976 TI - [Recommendations of the national AIDS committee]. PMID- 2972977 TI - [Expenditures and costs of diagnosis and therapy of HIV-infected patients]. PMID- 2972978 TI - [Multiyear observations of eye function in the miniaturist painters of Palekh]. PMID- 2972979 TI - [Mechanism of action of interferon-alpha 2 in hairy cells and lymphoblastoid cell lines: the significance of type I interferon receptors and RNA synthesis]. AB - Lymphoblastoid cells were subjected to Western Blot analysis for detection of Interferon-alpha-binding proteins. JOK-1 cells--a human hairy cell leukemia line- revealed three proteins with apparent molecular weights of 120, 100 and 32 kD, respectively. Down-regulation of the receptor was observed. A differential binding pattern of two proteins (100 and 85 kD) was observed in T-CLL, whereas no signal detection was achieved in B-CLL. Mononuclear cells from 6 patients with hairy cell leukemia and 6 patients with CLL were found to differ significantly in terms of nucleic acid precursor incorporation. PMID- 2972980 TI - Alterations of lens metabolism with experimentally induced cataract in rats. AB - Rat lenses with experimentally induced cataract (either by naphthalene or by streptozotocin) were analyzed biochemically. Both noxae had some effects in common. Water-soluble protein and aldose reductase activity decreased, and glucose-6-phosphate dehydrogenase, phosphofructokinase and glutathione reductase activity increased. A specific effect of streptozotocin was the rise in glucose, fructose and sorbitol. A specific effect of naphthalene was increased amounts of water-insoluble protein. PMID- 2972981 TI - Effect of alpha-human atrial natriuretic peptides on intraocular pressure in normal albino rabbits. AB - The effect of alpha-human atrial natriuretic peptide (alpha-hANP) on the intraocular pressure (IOP) was studied in normal rabbits. Dose response studies to IOP were performed after intravenous administration of alpha-hANP at 6.25, 12.5, 25.0, 50 and 100 micrograms/kg to the rabbits. The maximum reduction in IOP was observed to occur with the alpha-hANP 50 micrograms/kg dose. The time course of the pressure response to alpha-hANP were measured prior to and following intravenous administration of 50 micrograms/kg of alpha-hANP. The change in IOP and blood pressure showed the same pattern in the simultaneous record. Silver grains of 125I-alpha-hANP (10 microCi/g body weight) were detected in the ciliary processes of mice by radioautography after intravenous injection. The pupillary response was not tested in this experiment. The mechanism of IOP reduction was discussed. PMID- 2972983 TI - Antibody testing and counseling of dental patients at risk for human immunodeficiency virus (HIV) infection and associated clinical findings. AB - Two hundred six dental patients were tested between 1985 and 1987 for antibodies to human immunodeficiency virus (HIV) when a review of their medical histories revealed a high risk for infection. Serologic results are correlated with soft tissue and osseous findings recorded during routine head and neck and radiographic examination. Counseling recommendations for use in association with testing are outlined. A more active role for the dentist as a preventive agent is advocated to combat the spread of acquired immunodeficiency syndrome (AIDS). PMID- 2972982 TI - Histochemical characteristics of masseter and temporalis muscles after 5 weeks of maxillomandibular fixation--an investigation in Macaca mulatta. AB - This study describes the histochemical characteristics and cross-sectional areas of the superficial masseter and temporalis muscles in juvenile rhesus monkeys after 5 weeks of maxillomandibular fixation. Four juvenile male Macaca mulatta underwent mandibular surgery and 5 weeks of maxillomandibular fixation as part of a study of temporomandibular joint (TMJ) adaptations after condylar replacement. Immediately before the time the animals were killed (5 weeks postsurgically), biopsies of the masseter and temporalis muscles were obtained and submitted to histochemical analysis and calculation of muscle-fiber areas. The data were compared to histochemistry from 12 juvenile control Macaca mulatta. Significant decreases in mean cross-sectional area were exhibited in both type I (p less than 0.05) and type II (p less than 0.01) fibers in all muscles when compared to controls (n = 12). The ratio of type I to type II fibers, however, remained constant during maxillomandibular fixation in masseter and temporalis muscle samples, indicating no change in relative types of fibers. We conclude from this experimental investigation that (1) significant atrophy occurs in the temporalis and masseter muscles after 5 weeks of maxillomandibular fixation, and (2) this atrophy occurs in both type I and type II fibers, indicating that overall recruitment of the muscle (and not just of one fiber type of motor unit) was affected during fixation. PMID- 2972984 TI - Hereditary hemorrhagic telangiectasia: family study and review. AB - Hereditary hemorrhagic telangiectasia is reviewed, and the orofacial features of the condition are illustrated by details of an affected family and in particular a male patient who is now 63 years old. The nature of the disease from its insidious onset in the second and third decades to involve not only the mucous membranes but all cutaneous and visceral structures is noted, together with medical complications. Aspects of general and dental management are summarized. PMID- 2972985 TI - The prevalence of mitral valve prolapse in patients with Down's syndrome: implications for dental management. AB - Eighty-three noninstitutionalized patients with Down's syndrome, aged 9 to 55 years, were randomly selected to receive echocardiograms. Forty-one patients had echocardiographic findings indicative of mitral valve prolapse, and 15 of these patients lacked associated auscultatory findings. Because mitral valve prolapse can predispose patients to bacterial endocarditis after bacteremia-producing dental procedures, these findings suggest that if auscultatory findings alone are used, a significant number of patients with Down's syndrome who are at risk for endocarditis may not be currently identified in the course of routine clinical practice. PMID- 2972986 TI - [Health pedagogic foster families. A model for the rehabilitation of handicapped children from families at-risk]. AB - Rehabilitation of retarded children puts an enormous strain on parents. Families with various social risk factors are frequently not able to cope with these problems. Admission to institutes is a frequent consequence. Since 1982 47 mentally and psychologically handicapped children from families with various psychosocial problems were admitted to foster families. Foster parents were carefully selected and educated by a team of physicians, psychologists and social workers and given continuing supervision. Children were evaluated by medical and psychological testing: The majority of children showed significant improvement of performance in psychologic testing, alleviation in their psychosocial as well as medical problems. Numbers and duration of hospital admissions were substantially reduced. The overall condition of these children was markedly improved by foster families. PMID- 2972987 TI - Pain clinic #11. Physical exercise in the treatment of low back pain. Part III: A practical regimen of strengthening exercise. AB - The first and second articles in this three-part series reviewed the research evidence for physical exercise in the treatment of low back pain and outlined a specific and practical program of stretching exercise. In this final paper, we will present and illustrate the remaining component of a physical exercise regimen utilized in the treatment of a large population of patients suffering either nonsurgical or postsurgical low back pain. PMID- 2972988 TI - Bronchial compression by an enlarged left atrium in infants; a cause of hypovascularity of the left lung. AB - In three infants seen recently at our institution we noted signs of compression of the left main bronchus associated with enlarged left atria. None of our cases demonstrated the more usual signs of hyperinflation which are a hyperlucent lung field, depressed hemidiaphragm and mediastinal shift away from the affected side. In addition, hypoperfusion of the left lung was noted in each case. We believe that bronchial compression due to an enlarged left atrium, with consequent hypoxic vasoconstriction as a clinically significant entity, which is not well described and may be unappreciated in infants in whom the typical signs of hyperinflation are absent. PMID- 2972990 TI - [Disorders of the heart rhythm and conduction in adolescents (clinico epidemiologic research)]. PMID- 2972989 TI - IgE receptor-bearing lymphocytes in allergic and nonallergic children. AB - Using a monoclonal anti-human IgE receptor (Fc epsilon R) antibody, the percentage of Fc epsilon R(+) cells among peripheral blood lymphocytes in children with or without allergic disorders was determined. The percentage of Fc epsilon R(+) cells in 63 nonallergic children was 4.3 +/- 1.5%, which did not vary with age and was equal to that of adults (4.2 +/- 1.2%). Allergic younger children (0-2 yr) showed a significantly higher percentage of Fc epsilon R(+) cells (7.7 +/- 3.0%) than nonallergic younger children (0-2 yr) (4.0 +/- 1.3%, p less than 0.001). Similarly, in allergic younger children, serum IgE levels (geometric mean = 58.9 IU/ml) were also significantly higher than those of nonallergic younger children (geometric mean = 2.0 IU/ml) (p less than 0.01). A positive correlation between the percentages of Fc epsilon R(+) cells and serum IgE levels was observed (Spearman rank = 0.88, p less than 0.01] in eight allergic younger children (0-2 yr) with serum IgE levels higher than 100 IU/ml. The increase in the percentage of Fc epsilon R(+) cells in allergic younger children (0-2 yr) was not a secondary phenomenon caused by serum IgE because serum IgE levels in these children were much lower than the concentration at which IgE enhance Fc epsilon R expression on lymphocytes. In conclusion, Fc epsilon R(+) lymphocytes may play a regulatory role in IgE synthesis in allergic younger children (0-2 yr). PMID- 2972991 TI - [Laparoscopy in the diagnosis of acute appendicitis in children]. PMID- 2972992 TI - [Experience of a 6-month prophylactic course of multivitamins for children starting school from 6 years of age]. PMID- 2972994 TI - Sequence specificity of streptozotocin-induced mutations. AB - The isolation and characterization of streptozotocin (STZ)-induced mutations in the phage P22 mnt repressor gene is described. Cells carrying the plasmid-borne mnt gene were exposed to STZ to give 10-20 percent survival and at least an eleven-fold increase in mutation frequency. DNA sequence analysis showed that 50 of 51 STZ-induced mutations were GC to AT transitions, and one was an AT to GC transition. We have also compared the STZ mutational spectrum to that for N methyl-N'-nitro-N-nitroso-guanidine (MNNG). There are sites in the mnt gene which are mutated only by STZ; only by MNNG, or by both agents. Sites at which only STZ induced GC to AT transition mutations occur were in sequences that are pyrimidine rich 5' to the mutated site and purine rich 3' to the mutated site. Induction of mutations by both STZ and MNNG should be considered to maximize the number of mutable sites. PMID- 2972993 TI - Empirical estimation of protein-induced DNA bending angles: applications to lambda site-specific recombination complexes. AB - Protein-induced DNA bending is an important element in the structure of many protein-DNA complexes, including those involved in replication, transcription, and recombination. To understand these structures, the path followed by the DNA in each complex must be established. We have generated an empirical relation between the degree of bending and the altered electrophoretic mobility in polyacrylamide gels that allows estimation of protein-induced bends. This technique has been used to analyze 17 different protein-DNA complexes formed by six proteins including the four proteins involved in lambda site-specific recombination. The simplicity of this technique should make it useful in estimating angles for the construction of models of protein-DNA complexes and readily applicable to many systems where questions of higher-order structure are important for understanding function. PMID- 2972995 TI - Sequence of a transcribed Physarum genomic DNA fragment containing a cluster of different U-RNA sequences. PMID- 2972996 TI - Nucleotide sequence of a fungal plasmid-like DNA containing the mitochondrial ATPase subunit 6 gene. PMID- 2972997 TI - Synthesis of N-nicotinoyl-tryptamine (tryptamide). AB - N-Nicotinoyl-tryptamine was synthetized by acylation of tryptamine with mixed nicotinic anhydride. The synthesis of tryptamine via DL-tryptophan decarboxylation in cyclohexanol, in the presence of tetraline oxidation products as the catalyst, was described as well. PMID- 2972998 TI - Basic pharmacological properties of a novel antiinflammatory drug tryptamide. AB - In rats and mice the basic pharmacological properties of tryptamide (TRP), a novel antiinflammatory substance were studied. The LD50 doses were for male rats 1260 mg/kg ip or 8.5 g/kg po, for male mice: 1980 mg/kg ip or 9.3 g/kg po. TRP produced evident antiinflammatory effects of potency comparable with those of phenylbutazone when studied by means of carrageenin-induced rat paw oedema and prostaglandin synthetase activity in vitro. TRP reversed pyrogen-induced hyperthermia in rats, elicited analgesic effects in rats, but not in mice, prolonged the time of hexobarbital sleep in rats and inhibited locomotor activity in rats and mice. TRP has not elicited side effects on the circulatory system of rats and cats. It is concluded that TRP may undergo clinical trials as a potential antiinflammatory drug. During these trials the attention should be paid to possible central side effects the drug. PMID- 2973000 TI - The anti-inflammatory and analgesic activity of N-3-pyridoyltryptamine (tryptamide). AB - In this study we compared the antiinflammatory and analgesic properties of N-3 pyridoyltryptamine (tryptamide, TRP) with those of standard non-steroid anti inflammatory drugs (phenylbutazone, indometacin, piroxicam and ibuprofen). Antiinflammatory properties were investigated in the carrageenin-, serotonin-, kaolin-, formalin- and xylene-induced edema tests and in the cotton pellet granuloma test. The analgesic properties were studied in the writing, hot plate and electrical stimulation tests. In the majority of the applied tests TRP reveals the activity resembling this of phenylbutazone, and several times weaker than the effect of the remaining standard drugs. The ulcerogenic effect of TRP is very weak, and its effective doses are several times higher than these of the compared substances. Hence the therapeutic indices of TRP are several times higher than these of the other examined substances. The obtained results indicate that TRP is a putative anti-inflammatory and analgesic drug, much safer that other known substances with a similar pharmacological profile. PMID- 2972999 TI - Some pharmacological properties and subacute and chronic toxicity of tryptamide. AB - Studies on Albino Swiss mice and Wistar rats have demonstrated that tryptamide is less ulcerogenic than phenylbutazone, and markedly inhibits intestinal peristalsis. Both compounds have a similar tendency to accumulate in the body. Tryptamide produces a smaller hypotension and stimulates the respiratory amplitude to a lesser extent than phenylbutazone in a vivisectional experiment. Studies on the subacute and chronic toxicity have demonstrated that tryptamide administered orally (po) and intraperitioneally (ip) for 3 weeks, and orally for 3 months neither affects the body weight gain or the mass of internal organs, nor changes the locomotor activity; only in rats it disturbs the motor coordination. After ip administration tryptamide shows a moderate depressant effect on the bone marrow, evidenced by a decline in the blood hemoglobin content and the number of erythrocytes and blood platelets. As those changes were more pronounced after a 3 week than a 3-month administration, the observed effects are apparently reversible. PMID- 2973001 TI - Pharmacological investigations on anti-inflammatory activity of tryptamide (3 nicotinoyl-tryptamine). AB - The study concerned anti-inflammatory activity of a new tryptamine derivative - tryptamide. This preparation was effective in nonspecific inflammations and those involving immune mechanisms. Only postadjuvant inflammation appeared resistant to its effect. Its particular effectiveness was noted in the experimentally-induced iridocyclitis. Only in few tests, tryptamide was more effective than the reference drug phenylbutazone; in some cases the effect of both drugs was similar, but in the majority of cases tryptamide proved less effective. PMID- 2973003 TI - A spectrophotometric method for N-nicotinoyl-tryptamine (tryptamide) assay in the blood serum of rats. AB - A spectrophotometric method for N-nicotinoyl-tryptamine (tryptamide) (TR) assay in the blood serum of rats has been elaborated. Sensitivity of the method is approx. 1 microgram of TR/ml of blood serum, recovery approx. 76%. Using this method a preliminary pharmacokinetic study of TR was carried out in rats after ip or po administration of different doses of TR. It was found that the total amount of TR, expressed as corresponding AUC values, was nearly the same after both routes of administration of the same dose of TR. In addition, an effect of two solubilizing agents, 1% Tween 80 and gum arabic on TR (free base or hydrochloride) absorption from gastrointestinal tract of rats into general circulation has been investigated. Results of this study indicated the best absorption of TR when the substance, was given to rats as a hydrochloride suspended in 1% Tween 80 solution. PMID- 2973002 TI - A comparison of anti-inflammatory, analgesic and ulcerogenic action of tryptamide, ibuprofen and piroxicam. AB - Tryptamide, similarly to ibuprofen and piroxicam, shows anti-inflammatory and analgesic properties. Though tryptamide is much less potent than the standard drugs, its therapeutical indices are much more favorable. PMID- 2973004 TI - Influence of N-3-pyridoyltryptamine (tryptamide) on fetal development in rats and mice. AB - N-3-pyridoyltryptamine (Tryptamide, TRP) was examined for embryotoxic and teratogenic activities in rats and mice. Female Wistar rats and Albino Swiss mice were given TRP per os on days 6-14 of gestation. The rats were killed on day 21, and mice on day 18 of pregnancy. The general health and reproduction of females were evaluated, and fetuses were examined and processed to characterize external, visceral and skeletal malformations. TRP given to mothers in doses of 10 to 500 mg/kg po did not produce statistically significant embryotoxic or teratogenic effects in rats. It was not teratogenic also in mice but in this species the dose of 500 mg/kg produced embryotoxic effects. PMID- 2973005 TI - Angioplasty versus bypass surgery. Where do we stand today? PMID- 2973006 TI - Acute myocardial infarction. The primary care physician as gatekeeper. PMID- 2973007 TI - Irritant contact facial dermatitis due to nebulizer therapy. AB - A case is reported of a patient with long-standing, severe airflow obstruction requiring long-term nebulizer therapy developing a facial dermatitis in the area bounded by the nebulizer mask. The facial dermatitis seems to be the result of the combined irritancy of the nebulizer solutions and moisture, and prophylactic measures are suggested for patients requiring long-term nebulizer therapy. PMID- 2973008 TI - Electroacupuncture compared with paracetamol for acute low back pain. PMID- 2973009 TI - Effects of temperature on the in vitro binding of 3H-raclopride to rat striatal dopamine-D2 receptors. AB - The influence of temperature on the in vitro binding of 3H-raclopride to rat striatal dopamine-D2 receptors was investigated. The KD-values obtained in Scatchard plots were approximately 1.6 nM at temperatures between 15 degrees and 30 degrees. At 37 degrees the KD-value was found to be 3.0 nM, indicating a lower affinity without affecting the number of the receptors (Bmax). The rate of association of 3H-raclopride to the receptors was decreased with decreasing temperatures, and at 6 degrees more than 150 min. incubation was needed to reach the steady state level. From the association constants, the activation energy of the binding reaction was calculated to be 80 kJ/mol. The driving forces of the binding reaction was suggested to be a change in entropy at temperatures up to 30 degrees but a change in enthalpy at 37 degrees. PMID- 2973011 TI - [Does biliary dyskinesia exist? Arguments of perduodenoscopic manometry]. PMID- 2973010 TI - [Is the prevention of acute drug induced nephrotoxicity possible?]. PMID- 2973012 TI - [Treatment of refractory congestive cardiac insufficiency by continuous peritoneal dialysis. Long-term course]. AB - Continuous and progressive fluid removal was performed by continuous peritoneal dialysis in 19 patients (14 men, 5 women, mean age 60.7 years) suffering from refractory congestive heart failure. All patients were in NYHA class IV and had a life-threatening fluid overload. Twelve had normal renal function or functional renal failure, and 7 had organic renal failure. The continuous peritoneal dialysis technique with a high K+ concentration in the dialysate did not raise any particular problem. Mean survival of the whole group was 7.1 months. This figure rose to 16 months (rang 2-51 months) in the patients who survived at the end of the first month. Sixteen out of 19 patients eventually died, most of them suddenly and probably of ventricular arrhythmia. Nine patients were discharged on continuous ambulatory peritoneal dialysis and remained on it for 2 to 48 months. Drug therapy could be reduced in all cases. Dialysis was discontinued in 5 patients without organic renal failure who thereafter survived for a mean period of 5 months. Functional improvement and duration of survival were uncorrelated with heart disease, fluid overload, initial renal function or initial left ventricular ejection fraction. A cardiothoracic ratio greater than 0.70 and the need for mechanical ventilation seemed to be of poor prognosis. Such results justify the pursuit of this study before defining selection criteria. PMID- 2973013 TI - [Influence of oral taurine supplementation on the intraduodenal concentration and conjugation of bile acids in full-term newborn infants]. AB - We studied the effects of oral taurine supplementation on bile acids conjugation and duodenal bile salt concentrations in infants. Seventeen infants receiving enteral artificial nutrition were investigated. At the beginning of the study they were 6 to 14 weeks old, in good nutritional state, without malabsorption, protein-losing enteropathy and liver or infectious diseases. After at least 8 days of a stable, taurine-free regimen the infants received oral taurine supplementation (36-45 micromol/kg.24 h) for 8 days. Bile acids were measured before and after each supplementation period in bile samples obtained by duodenal tubing, using enzymatic methods and colorimetry. According to the initial plasma taurine levels before supplementation, the infants were divided into two groups: I) plasma taurine levels less than 60 mumol/l (mean 47 +/-5 mumol/l, n = 8); II) plasma taurine levels greater than 70 mumol/l (mean 77 +/- 2 mumol/l, = 9). After 8 days of taurine supplementation a significant increase of plasma and urinary taurine (P less than 0.01),total duodenal bile salt concentrations (P less than 0.05), total duodenal tauroconjugates (P less than 0.05),taurocholate (P less than 0.01), taurochenodeoxycholate (P less than 0.05), and glycocholate (P less than 0.01), duodenal concentrations, and a significant decrease of the glycoconjugate/tauroconjugate ratio (P less than 0.05), were observed, but only in group I. in group II infants we only noted a significant increase of urinary taurine (P less than 0.01), and of duodenal total tauroconjugates (P less than 0.05). This study shows that the biliary effects of an oral taurine supplementation depends on taurine status and that in taurine-depleted infants intakes of exogenous taurine higher than 45 mumol/kg. 24 h are perhaps necessary for optimal bile salt effects. PMID- 2973014 TI - [Transfer of measles immunoglobulins and antibodies from mother to child in Africa and Europe]. AB - In a series of 50 mother-child pairs from Paris the titers of measles antibodies in cord blood were 1.4 times higher that those measured in the mother's blood, but this ratio was only 0.92 in 116 African mother-child pairs from Libreville (Gabon). Similarly, the ratio of immunoglobulin G levels in cord blood to those of the mother's blood was higher in Paris (1.34) than in Libreville (0.88). This limitation in active transfer of total of specific immunoglobulins is due to their high levels usual in African mothers, and this is not without repercussions on the immune defence mechanisms of African newborns. PMID- 2973015 TI - [Tamm-Horsfall protein]. AB - Tamm-Horsfall protein is the most abundant protein in normal urine and the main component of urinary casts, but its precise physiological role remains to be elucidated. The interest raised by this protein has recently been boosted by the finding that its gene is identical with that of uromodulin, a glycoprotein purified from urine of pregnant women and endowed with immunosuppressive activity. In this review, are analyzed successively: the structure and physico chemical properties of Tamm-Horsfall protein which is remarkable by its capacity for gel formation, its localization along the nephron where it is synthesized by cells of the thick ascending limb of Henle's loop, its extra-renal distribution apparently restricted to the serum and its potential physiological and pathological roles. Studies of renal biopsies with specific antibodies may provide information on the part taken by tubular obstruction in the development of renal failure. PMID- 2973016 TI - [Screw fixation of the odontoid process]. AB - Fractures of the odontoid process of the axis create problems of specific osteosynthesis. The different techniques described so far require an external support and limit the movements of the cranio-cervical joint. These 2 drawbacks are avoided by direct screw fixation of the odontoid process in cases where conditions are favourable to this procedure. PMID- 2973018 TI - [Isolated cerebral nocardiosis. A case]. PMID- 2973017 TI - [Focal nodular myositis simulating a sural thrombophlebitis. Association with cryoglobulinemia with polyneuropathy and glomerular disease]. PMID- 2973019 TI - [Coronary localization of Horton's disease]. PMID- 2973020 TI - [Lead poisoning caused by glazed pottery]. PMID- 2973021 TI - [Idiopathic hemochromatosis associated with Klinefelter's syndrome]. PMID- 2973022 TI - [Paralysis after surgery of the pelvis]. PMID- 2973023 TI - [Value of ketoprofen in obstetrical surgery]. PMID- 2973024 TI - [Hypokalemia in a patient treated with theophylline. A case]. PMID- 2973025 TI - [Association of Hansen's disease and human deficiency virus infection. 2 cases]. PMID- 2973026 TI - [Acute regressive myelopathy in infection with human immunodeficiency type 1 virus]. PMID- 2973027 TI - [Choice of immunosuppression in the graft of kidney]. PMID- 2973028 TI - [New treatments of pain in digestive tumor pathology]. PMID- 2973029 TI - [Cardiac toxicity of 5-fluorouracil. Review of the literature, 5 new cases]. AB - The toxic effects of 5-fluorouracil - an antimitotic drug widely used in the treatment of cancer - mainly affect the digestive tract and the blood. The 5 new cases presented here may be added to the 57 cases of cardiotoxicity reported in the literature. The clinical manifestations always consist of constrictive chest pain resembling angina pectoris and associated with disorders of repolarization on electrocardiographic tracing recorded during the painful episodes. The outcome is usually favourable after discontinuation of treatment, but it may also be fatal. Reintroduction of the drug in 28 patients resulted in myocardial necrosis in 4 cases and in death due to cardiogenic shock in 4 cases. The frequency of such side-effects does not seem to be influenced by age, sex, route of administration and previous pathology including cardiovascular diseases. They usually occur at the beginning of treatment irrespective of the dose administered. The mechanism through which 5-fluorouracil exerts its cardiotoxic effects is unknown, and 3 hypothesis have been put forward: an immunoallergic reaction after a sensitization period, a coronary spasm directly induced by the drug or due to the release of a vasopressive substance, or a direct toxic effect on the myocardium and pericardium. PMID- 2973030 TI - [Comparison between treatments of severe forms of familial hypercholesterolemia by total plasma exchange and selective removal of low density lipoproteins (LDLapheresis)]. AB - A plasma exchange program for familial hypercholesterolaemia was started in 1982. Ten patients aged from 7 to 58 years were progressively included: 3 had an heterozygous form of the disease with ischaemic heart disease; 3 had an homozygous form with defective low density lipoprotein receptor activity, 4 had a receptor-negative homozygous familial hypercholesterolaemia and had previously undergone portacaval shunt. During total plasma exchange against human albumin (470 sessions in 9 patients) low density lipoprotein cholesterol values, but also high density lipoprotein cholesterol values, decreased by 40 per cent. More recently, 5 patients had selective low density lipoprotein absorption on dextran sulfate column (Liposorber); 90 exchanges were performed. High density lipoprotein cholesterol values decreased by 55 per cent and high density lipoprotein cholesterol values by only 27 per cent. The patients' attitude to treatment was excellent, with less fatigue and better compliance. PMID- 2973031 TI - [Dissolution of gallbladder lithiasis with methyl-tert-butyl-ether. Preliminary results: 9 cases]. AB - Nine patients with gallstones were treated by injection of methyl-tert-butyl ether (MTBE) into the gallbladder. Complete or incomplete dissolution was observed in 5 of the 9 patients. The 4 failures were due to 2 main factors: poor selection of the patients (2 of them had pigment gallstones) and inadequate mixture with the solvent floating above the bile and gallstones. In addition, 4 complications were noted, all of them being resolved by medical treatment. A better selection of the patients and a more efficient technique of MTBE injection should improve these results. PMID- 2973032 TI - [Peritoneo-gluteal suppurative hidradenitis]. AB - Perineo-gluteal hidradenitis suppurativa, or Verneuil's disease, consists of infected and more or less fistulized lesions unconnected with the anal canal, dermo-epidermal nodule and diffuse subcutaneous infiltration. The disease is chronic with acute episodes and periods of stabilization and it interferes with the patient's daily life. The lesions originate in the apocrine sweat glands which are predominantly located in the inguinal, perineal and gluteal regions but may also be found in the axillary, areolar and periumbilical regions. The cause of the disease is unknown, but it has been attributed to an anaerobic Streptococcus, to hormonal factors or to maceration or dysembryoplasia. Treatment consists of wide excision of the lesions followed by a network of skin grafts after a granulation period. Isotretinoin has not yet been proved effective. PMID- 2973033 TI - [Chronic alcohol-tobacco poisoning: towards an early detection of optic neuropathy]. AB - Forty-two patients (33 men, 9 women) with normal visual acuity admitted to an alcoholism rehabilitation programme were evaluated for colour vision using the automatized Fransworth-Munsell test (ChromopsR) and for central visual field using Friedmann's analyser. An early colour blindness without axis, i.e. a lack of colour discrimination, was often found. At the same time, in the more intoxications perimetric defects appeared in the 30 degrees area first centrally, then followed by arcuate superior scotomas. PMID- 2973034 TI - [Lymphocyte alveolitis and paraplegia associated with HTLV-I]. PMID- 2973035 TI - [Major thrombocytopenia related to HIV1 virus treated with zidovudine]. PMID- 2973036 TI - [Deficiency in immunoglobulin G subsets associated with primary infectious pneumopathies in adults]. PMID- 2973037 TI - [Pulmonary aspergillomas and pancreatic localization in disseminated aspergillosis]. PMID- 2973038 TI - [Hypercalcemia associated with hyperleukocytosis in epidermoid carcinoma of the cervix uteri]. PMID- 2973039 TI - [Permanent deficiency of the visual field following migraine]. PMID- 2973040 TI - [The prognosis of hemolytic-uremic syndrome may be favourable in the elderly]. PMID- 2973041 TI - [Occurrence of multiple myeloma 5 years after renal transplantation]. PMID- 2973043 TI - [Analgesic action of sodium diclofenac: behavioral and electrophysiological study in normal and polyarthritic rats]. AB - In a behavioural study, the effects of intravenous diclofenac in a range of doses were determined by measuring vocalization thresholds to paw pressure in normal and Freund's adjuvant-induced arthritic rats. In normal rats, 6 and 9 mg/kg diclofenac produced an increase in vocalization thresholds (mean threshold values were respectively 135.67 +/- 3.30 per cent and 157.41 +/- 4.62 per cent of the control at 30 min; n = 9 for each group), no effect resulting with 3 mg/kg. In arthritic rats, 3, 6 and 9 mg/kg diclofenac induced a clear analgesic effect (mean threshold values being 172.22 +/- 4.26 per cent, 201.76 +/- 4.76 per cent and 222.33 +/- 5.10 per cent respectively of the control at 25 min, n = 9 for each group), no effect being observed with 1.5 mg/kg. An electrophysiological study performed on arthritic rats considered the effect of 6 mg/kg diclofenac upon the neuronal ventrobasal thalamic responses induced by mode-rate stimulation of an inflamed joint. The responses were depressed by about 50 per cent, 20 minutes after the injection. These results show that diclofenac displays a direct effect on pain processing; this effect is more potent in arthritic rats than in normal rats. PMID- 2973042 TI - [Can the indications of conservative radiosurgical combination in the treatment of breast cancer be extended?]. AB - In the hope that women with breast cancer unsuitable for primary tumorectomy would benefit from a conservative treatment, we tried to reduce the lesional volume by an initial irradiation of 45 Gy administered in 25 fractions over 35 days. This treatment was applied to 111 patients (69 T2; 42 T3). In 44 cases (40 per cent), tumoral regression enabled a secondary tumorectomy to be performed. In 16 cases (14 per cent), the tumour totally disappeared, and an additional irradiation by endocurietherapy in intermediate dosage could be given. In 51 patients (46 per cent), the tumoral response was poor and the patients underwent, depending on their wishes, either mammectomy (9 cases; 8 per cent) or high-dose endocurietherapy (42 cases; 38 per cent). Analysis of tumoral regression according to size, histological type and mammographic image provided for a better identification of good responders. The cosmetic and oncological results of this preliminary study are satisfactory, with a 95 per cent local control at 5 years after secondary tumorectomy. PMID- 2973045 TI - [Juvenile multiple myeloma. A Nigerian case]. AB - One case of juvenile multiple myeloma of the IgG-kappa type, with lytic lesions of the skull, is reported in a 14-year old Nigerian Fulani girl. The outcome was rapidly fatal. This disease is extremely rare: only 2 other documented cases (2 girls aged 13 and 12) have been found in the literature, under the age of 15. PMID- 2973044 TI - [Hematomas of the rectus abdominis muscle. 27 cases]. AB - A series of 27 haematomas of the rectus abdominis muscle is reported; 10 occurred under anticoagulants, 6 were spontaneous, 5 followed a trauma, and 6 were consecutive to a surgical procedure. The clinical picture was subacute in 20 cases, with abdominal pain and extensive ecchymosis, acute in 4 cases, and reduced to an abdominal mass in 3 cases. Since 1977, ultrasonography has been of considerable help in the diagnosis, which was found to be accurate in 5 out of the 10 cases seen before its introduction, and in 13 out of the 14 cases where it was used among the 17 cases recorded later. Nineteen patients were operated upon for the following reasons: 4 for cardiovascular collapse, 3 because no diagnosis had been made, 8 in order to confirm the diagnosis, 1 because of an increase in size of the haematoma and 3 owing to infection. In the 8 patients who did not undergo surgery, the haematoma followed a favourable course. The therapeutic approach must be chosen according to the patient's condition and to the size, side effects and course of the haematoma. PMID- 2973046 TI - [Indwelling endoluminal balloon. An uncommon method of peroperative hemostasis]. AB - A case of ruptured false aneurysm of the left common artery is reported, in which intra-operative haemostasis of the left internal artery proved particularly difficult. Permanent haemostasis was achieved by inserting an inflatable balloon intraluminally at the origin of that artery and leaving it in situ. PMID- 2973047 TI - [Hypothermia in Gayet-Wernicke syndrome and subclinical centropontine myelinolysis]. PMID- 2973048 TI - [Study of a possible pristinamycin-antipyrine interaction]. PMID- 2973050 TI - [Increase of cases of tuberculosis in 3 hospitals in Paris]. PMID- 2973051 TI - [Uretero-hydronephroses disclosing infection of aortic prosthesis]. PMID- 2973049 TI - [Anicteric cholestasis disclosing Horton's disease]. PMID- 2973052 TI - [Non-traumatic rhabdomyolysis in Staphylococcus aureus septicemia]. PMID- 2973053 TI - [A new echographic sign: left intraventricular isovolumetric relaxation flow]. PMID- 2973054 TI - [Association of primary hyperparathyroidism and secondary polycythemia]. PMID- 2973055 TI - [Cytological test of nipple discharge]. PMID- 2973056 TI - [Factitious thyrotoxicosis with periodic paralysis]. PMID- 2973057 TI - Cleavage of the cII protein of phage lambda by purified HflA protease: control of the switch between lysis and lysogeny. AB - The activity of the cII protein of phage lambda is probably the critical controlling factor in the choice of the lytic or lysogenic pathway by an infecting virus. Previous work has established that cII activity is regulated through the turnover of cII protein; the products of the hflA and hflB loci of Escherichia coli are needed for a degradative reaction, and lambda cIII functions in stabilizing cII. By using the cloned hflA locus, we have purified a cII cleaving enzyme that we term HflA. Purified HflA contains three polypeptides; at least two of the subunits are products of the hflA region, and the third is probably a cleavage product of the larger of these two hflA-encoded polypeptides. The HflA protease activity cleaves cII to small fragments. We conclude that the switch between lambda developmental pathways involves regulated cleavage of cII by the specific protease HflA. PMID- 2973058 TI - Bafilomycins: a class of inhibitors of membrane ATPases from microorganisms, animal cells, and plant cells. AB - Various membrane ATPases have been tested for their sensitivity to bafilomycin A1, a macrolide antibiotic. F1F0 ATPases from bacteria and mitochondria are not affected by this antibiotic. In contrast, E1E2 ATPases--e.g., the K+-dependent (Kdp) ATPase from Escherichia coli, the Na+,K+-ATPase from ox brain, and the Ca2+ ATPase from sarcoplasmic reticulum--are moderately sensitive to this inhibitor. Finally, membrane ATPases from Neurospora vacuoles, chromaffin granules, and plant vacuoles are extremely sensitive. From this we conclude that bafilomycin A1 is a valuable tool for distinguishing among the three different types of ATPases and represents the first relatively specific potent inhibitor of vacuolar ATPases. PMID- 2973060 TI - T helper cell subsets require the expression of distinct costimulatory signals by antigen-presenting cells. AB - We examined the ability of macrophages and B cells to function as antigen presenting cells (APCs) for murine TH1 and TH2 cloned T helper cell lines. Antigen presented by concanavalin A-elicited peritoneal macrophages or resting splenic B cells stimulated antigen-dependent proliferation of both T helper subsets. Paraformaldehyde fixation of the APCs following different conditions of activation indicated differential requirements for costimulatory signals by TH1 and TH2 cells. TH2 proliferative responses were strictly dependent on APC expression of IL-1. TH1 proliferation was dependent on APC expression of a non-IL 1 costimulatory signal present on freshly isolated macrophages and on splenic B cells activated with anti-immunoglobulin plus interferon gamma. PMID- 2973059 TI - Presence of laminin-binding proteins in trichomonads and their role in adhesion. AB - Adhesion is regarded as an important feature in the pathogenesis of various microorganisms. Ability to recognize extracellular matrix proteins, such as laminin or fibronectin, has been correlated with invasiveness. We report that laminin enhances the adhesion of the parasitic protozoa Trichomonas vaginalis and Tritrichomonas foetus to a polystyrene substrate and to the surface of epithelial cells (Madin-Darby canine kidney cell line) in vitro. The enhancement was higher for T. vaginalis than for T. foetus. Addition of anti-laminin antibodies to medium significantly inhibited the adhesion of parasites to polystyrene substrate. Indirect immunofluorescence and transmission electron microscopy of replicas of the parasite's surface labeled with antibody-gold complexes showed laminin-binding sites distributed over the parasite surface. Iodinated P1 fragment of laminin, which retains the laminin-binding site, binds saturably to the parasite surface with a Kd of 19.5 nM, for about 3 X 10(5) binding sites per cell. Immunoblotting analysis of whole parasite extracts showed that a protein of 118 kDa is responsible for laminin binding. PMID- 2973062 TI - Molecular dynamics in the membranes of helper T cells. AB - We provide evidence that redistributions and interactions of integral proteins in the fluid membranes of helper T (Th) cells may play important roles in Th-cell activation. A particular monoclonal antibody, 3D3, directed to a clonotypic determinant on the T-cell receptor (TCR) of the cloned Th-cell line D10, had previously been shown to be distinctively capable of directly activating D10 cells at low concentrations. We demonstrate here by immunofluorescence experiments that it is also distinctively able itself to produce a clustering (capping) of the TCRs on the D10 cell surface. Simultaneously, by means of double immunofluorescence experiments, we find that the 3D3-induced clustering of the TCRs distinctively produces a co-clustering of the accessory molecule CD4 with the TCR clusters, although the CD4 and TCR molecules are normally independent of one another in the D10 cell membrane. These results, and related ones previously obtained from studies of the interactions of D10 Th cells with antigen-presenting cells, are analyzed to suggest that the membrane clustering of TCRs and the induced TCR-CD4 interactions are critical to the signaling events in Th-cell activation. PMID- 2973061 TI - Peptide-mediated modulation of T-cell allorecognition. AB - Antigen-specific helper T cells recognize a complex of peptide antigen and class II major histocompatibility complex (MHC) gene products. Whether T cells recognize MHC class II alloantigen by a similar mechanism or the native conformation of MHC molecules themselves has yet to be determined. The demonstration that peptide antigens bind directly and specifically to class II molecules has allowed us to examine the influence of foreign peptide binding on T cell recognition of allogeneic MHC molecules. We report here that an immunodominant, HLA-DR1-restricted peptide of influenza virus hemagglutinin (HA residues 306-320) is able to modulate the recognition of alloantigen by human DR1 specific T-cell clones. For some T-cell clones, but not all, the HA peptide inhibited allorecognition in a dose-dependent manner. However, in one instance, the proliferative response to alloantigen was enhanced in the presence of HA peptide. These results suggest that the specificities of T-cell responses to allogeneic MHC molecules are heterogeneous, which may be influenced by different peptides occupying the class II MHC combining site and by the diversity of antigen-specific receptors of T lymphocytes recognizing the same MHC/peptide complex. PMID- 2973063 TI - Mapping of the gene encoding the beta-amyloid precursor protein and its relationship to the Down syndrome region of chromosome 21. AB - The gene encoding the beta-amyloid precursor protein has been assigned to human chromosome 21, as has a gene responsible for at least some cases of familial Alzheimer disease. Linkage studies strongly suggest that the beta-amyloid precursor protein and the product corresponding to familial Alzheimer disease are from two genes, or at least that several million base pairs of DNA separate the markers. The precise location of the beta-amyloid precursor protein gene on chromosome 21 has not yet been determined. Here we show, by using a somatic cell/hybrid-cell mapping panel, in situ hybridization, and transverse-alternating field electrophoresis, that the beta-amyloid precursor protein gene is located on chromosome 21 very near the 21q21/21q22 border and probably within the region of chromosome 21 that, when trisomic, results in Down syndrome. PMID- 2973064 TI - Manipulation of stretch-induced atriopeptin prohormone release and processing in the perfused rat heart. AB - Atriopeptin (AP) is stored as the prohormone AP-126 [atrial natriuretic factor-(1 26)] in atrial granules. Cultured atrial myocytes synthesize and release only prohormone into the medium. HPLC analysis of the coronary venous effluent of media from perfused rat hearts subjected to right atrial stretch indicated the presence of the C-terminal mature hormone AP-28 [atrial natriuretic factor-(99 126)] and little or no prohormone. Absence of calcium from the perfusion medium increased total AP release and surprisingly blocked the proteolytic cleavage of the prohormone. Similarly, addition of the proteolytic inhibitor aprotinin to the perfusion medium suppressed the processing of the endogenous AP-126 released by atrial stretch. Aprotinin would be restricted to the extracellular space, which is therefore implicated as the site of prohormone processing. This suggestion was validated by the demonstration that the perfused rat heart could readily cleave exogenous prohormone to mature hormone, a process blocked by aprotinin. Hypothetically, the stimulus-release-processing event initiated by atrial stretch may require the concerted action of the synthetic cell (i.e., atrial myocyte) and a processing cell or site (e.g., the adjacent atrial mesenchymal cell) for the production of the mature AP-28, which is the circulating molecular form of this endocrine system. PMID- 2973065 TI - Localization of chitin synthetase in cell-free homogenates of Saccharomyces cerevisiae: chitosomes and plasma membrane. AB - We describe an improved method for fractionating cell-free extracts of Saccharomyces cerevisiae to separate its membranous components by a combination of isopycnic and velocity sedimentations. These procedures were used to examine the subcellular distribution of chitin synthetase (chitin-UDP acetylglucosaminyltransferase; EC 2.4.1.16) in homogenates from exponentially growing walled cells of a wild-type strain of yeast. Chitin synthetase (Chs1) activity was mainly found in two distinct vesicle populations of nearly equal abundance but with markedly different buoyant densities and particle diameters. One population contained 45-65% of the total chitin synthetase and was identified as chitosomes because of microvesicular size (median diameter = 61 nm) and characteristic low buoyant density (1.15 g/ml); it also lacked 1,3-beta-glucan synthetase activity. The second population (35-55%) was identified as plasma membrane because of its high buoyant density (1.22 g/ml), large vesicle size (median diameter = 252 nm), and presence of vanadate-sensitive ATPase. This fraction cosedimented with the main peak of 1,3-beta-glucan synthetase. A third, minor population of chitin synthetase particles was also detected. Essentially all of the chitin synthetase in the two vesicle populations was zymogenic; therefore, we regard these vesicles as precursors of the final active form of chitin synthetase whose location in the cell has yet to be unequivocally determined. PMID- 2973068 TI - Natriuresis of volume expansion and atrial factors: contrasts and comparisons. PMID- 2973066 TI - Characterization and expression of the murine CD3-epsilon gene. AB - The receptor for antigen on the surface of T lymphocytes consists of a variable disulfide-bridged hetero-dimer (TCR-alpha/beta or -gamma/delta) associated with invariant CD3 proteins (CD3-gamma, -delta, -epsilon, and -zeta). The genes coding for the CD3 proteins are expressed in the earliest recognizable thymocytes, preceding the rearrangement and expression of the TCR genes. The isolation, characterization, and in vitro expression of the murine CD3-epsilon gene, as reported here, represent obligatory steps toward our understanding of the complex rules that govern T-cell-specific gene expression. The CD3-epsilon gene was transcribed from a non-TATA promoter and consisted of eight exons, two of which were unusually small (18 and 15 base pairs). The transmembrane exon was found to be homologous to the transmembrane exons of the CD3-gamma and CD3-delta genes. In transient-transfection experiments, a genomic fragment comprising 4 kilobases of upstream sequence and extending into the second exon sufficient to drive the expression of a reporter gene in murine T cells. PMID- 2973067 TI - CD45 regulates signal transduction and lymphocyte activation by specific association with receptor molecules on T or B cells. AB - Evidence is presented that the leukocyte common antigen CD45 can regulate both signal transduction by lymphocyte receptor molecules and T- and B-cell proliferation in a manner dependent on specific interactions between these receptors on the cell surface. Formation of homoaggregates of CD3, CD2, or CD28 on the surface of T cells induced by crosslinking with monoclonal antibodies (mAbs) results in an increase in cytoplasmic free calcium concentration ([Ca2+]i). This increase in [Ca2+]i was abolished when these receptors were crosslinked to CD45 on the cell surface. In contrast, the increase in [Ca2+]i induced by formation of homoaggregates of CD4 was strongly amplified when CD4 was coupled to CD45. T-cell proliferation initiated by immobilized anti-CD3 was inhibited by anti-CD45 or anti-CD45R when immobilized on the same surface, but not when in solution. Similarly, proliferation after stimulation of the CD2 and CD28 receptors was inhibited when a CD45 mAb was crosslinked to either CD2 or CD28 mAbs, but not when a CD45-specific mAb was bound to the cell surface separately. In B cells, the increase in [Ca2+]i and resulting proliferation induced by crosslinking either the CD19 or Bgp95 receptors was inhibited by coupling these molecules to CD45. Thus, CD45 appears to modify other cellular receptors functionally when brought into close physical association with them. The homology of the CD45 conserved cytoplasmic domains with a major human placental protein tyrosine phosphatase suggests that the effects of CD45 described here result from alterations in the phosphorylation state of tyrosyl residues in membrane-associated proteins. PMID- 2973071 TI - [Prevention of low back pain]. PMID- 2973070 TI - A biochemical analysis of the effects of arachidonic acid on sarcoplasmic reticulum function. AB - Discontinuous sucrose gradients were used to determine the degree of association between arachidonic acid and sarcoplasmic reticulum vesicle membranes. Fraction analyses showed that arachidonic acid migrated to a different region of the sucrose gradient in the presence of sarcoplasmic reticulum membranes. This could suggest that arachidonic acid was complexed into the membranes. Arrhenius curves representing the temperature dependency of Ca2+-Mg2+-ATPase activity and calcium uptake in the presence and absence of arachidonic acid were constructed. The activation energy for ATPase did not change significantly due to the presence of arachidonic acid. The curve representing control calcium uptake did not show a discontinuity. However, the curve representing calcium uptake in the presence of arachidonic acid showed discontinuities at 18 degrees C and 21 degrees C. Activation energy increased sharply between these temperatures. The results suggest that arachidonic acid reached the critical micellar concentration between these temperatures. Enthalpy decreased in the presence of arachidonic acid. This observation could suggest a transition of the protein-phospholipid complex to a less rigid state since decreased order in the membrane would decrease the energy barrier for activation of ATPase. PMID- 2973069 TI - Selected advances in therapy of urologic cancer--luteinizing hormone releasing hormone (LHRH) and biologic response modifier (BUR). PMID- 2973072 TI - The concept of "marginality" applied to psychosomatic medicine. PMID- 2973073 TI - [Social interaction in patients with chronic backache]. PMID- 2973074 TI - Assay of radiation effects in mouse skin as expressed in wound healing. AB - The effect of 150 kVp X irradiation on the healing of full depth surgical wounds in the lower dorsal skin of the mouse was assayed by measuring the wound strength of seven 2-mm-wide segments along each wound. The strength of unirradiated wounds increased with time in two phases: during the first 2 weeks it reached nearly half of the values recorded from unwounded skin, after which the rate of increase slowed for at least 2 weeks before beginning a second increase. By 150 days, the breaking strength of the wound was about 80% of that of unwounded skin. A single dose of 18 Gy prior to wounding reduced the strength of the wounds to about one third to one-half that of an unirradiated wounds within the 3 months of follow up. The effect of irradiation on wound strength did not change as the interval between exposure and wounding was increased to 2 months but decreased slightly when this interval was extended to 3 months. When the healing wound was irradiated within 5 days of surgery, the effect on healing was about the same as with preirradiation; if irradiation was delayed for 12 days after wounding the second phase of healing was only postponed and the wound strength ultimately approached the values recorded from unirradiated wounds. The wound strength of skin preirradiated by X rays and assayed 14 days after wounding showed a clear sigmoid dose response with a threshold between 8 and 10 Gy and a plateau at the maximum effect above 20 Gy. The persistence for at least 3 months of the effect of radiation on wound healing suggests that the tissues involved in the healing process are normally proliferating slowly. The accelerated expression of radiation injury through surgical wounding permits the early quantification of the radiation response of tissues that would normally be delayed in their expression of radiation damage. PMID- 2973076 TI - [Percutaneous transluminal angioplasty of the brachiocephalic vein in sarcoidosis with superior inflow obstruction]. AB - A case of sarcoidosis is reported, which began 25 years ago. As a late complication the patient developed compression syndrome of the superior caval vein, which was caused by swelling of the lymph nodes. After surgical treatment, which was carried out by removing the regional lymph nodes and implantation of a dacron prosthesis, stenosis occurred within the proximal part of the prosthesis, which was treated by the PTA technique according to the Dotter method. PMID- 2973075 TI - Correlation between PLD repair capacity and the survival curve of human fibroblasts in exponential growth phase: analysis in terms of several parameters. AB - Published data on the in vitro radiosensitivity of 46 nontransformed fibroblasts of different genetic origins studied in plateau phase with immediate or delayed plating were used to investigate to what extent potentially lethal damage repair capacity is related to intrinsic radiosensitivity (i.e., irradiated in exponential growth phase). While most of the survival curve analysis is conducted in terms of D0, Dq, and the mean inactivation dose D, some of the data are also discussed in terms of the linear-quadratic model parameter alpha. Using D it is shown that: (i) the radiosensitivity of human fibroblasts in exponential growth phase does not significantly differ from that of plateau-phase fibroblasts with immediate plating; (ii) the radiosensitivity of plateau-phase cells with delayed plating is correlated to the radiosensitivity of cells with immediate plating: the more radioresistant the cell strain in exponential growth phase, the higher its repair capacity; (iii) the repair capacity of the cell strains is related to their genetic origin. In conclusion, we suggest that the survival curve of growing cells depends on the repair capacity of the cells. PMID- 2973077 TI - [Dilatation of the supra-aortic arteries--a catheter with blockade and rinsing potential]. AB - Percutaneous transluminal angioplasty (PTA) of supra-aortic arteries is not generally accepted, mainly because of the fear of brain embolism. The design of a new catheter offers four conduits and permits (1) guidewire conduct, (2) tip occlusion; (3) PTA; and (4) rinsing between occluded tip and dilational balloon. Thus, any particles that might otherwise cause brain embolism could be washed out after PTA and before opening of the tip occlusion. The prototype has already been manufactured. The catheter is easy to handle. PMID- 2973078 TI - Peripheral artery atherectomy: description of technique and report of initial results. AB - Percutaneous transluminal atherectomy has been developed for treatment of peripheral artery stenoses. The atherectomy catheter is inserted through a sheath, and the resection window of the catheter is positioned adjacent to the vascular stenosis. The balloon is inflated, and the motor-driven cutting blade advanced. The balloon is then deflated, the catheter withdrawn, and the atheromatous material, which resembles the resected material of an endarterectomy, removed from the catheter. This process is repeated until the resection provides an adequate lumen. To date, 12 arterial lesions (three common iliac, two external iliac, four superficial femoral, and three popliteal artery) in ten patients have been resected with excellent angiographic results. The conditions of seven patients who underwent atherectomy for relief from claudication were improved by the criteria of ankle/arm ratios and/or claudication distance. Three patients successfully underwent atherectomy for limb salvage. More data on long-term patency and restenosis rates are needed before the ultimate role of atherectomy in the management of peripheral artery disease can be determined. PMID- 2973079 TI - Cutaneous reaction to contrast material. AB - Two children are described who developed an apparent cutaneous contact reaction to contrast material in urine. Both children had undergone uneventful voiding cystourethrography with diatrizoate meglumine injection USP 18% followed by intravenous urography with diatrizoate meglumine injection USP 60%. Approximately 1 hour after urography cutaneous bullae and surrounding erythema of the buttocks (one case) or foreskin (one case) were noted. This reaction resembled a superficial chemical burn. PMID- 2973080 TI - A new challenge to radiologists. PMID- 2973081 TI - Who should blow up balloons in arteries? PMID- 2973082 TI - Responses of muscle spindles depend on their history of activation and movement. PMID- 2973083 TI - Influence of adrenaline and hypoxia on rat muscle receptors in vitro. PMID- 2973084 TI - The synthesis of a novel thromboxane receptor antagonist 4(Z)-6-(2-o-chlorophenyl 4-o-hydroxyphenyl-1,3-dioxan-cis-5-yl) hexenoic acid ICI 192605. AB - The synthesis and summary pharmacology of a novel thromboxane receptor antagonist 4(Z)-6-(2-o-chlorophenyl-4-o-hydroxyphenyl-1, 3-dioxan-cis-5-yl) hexenoic acid (3) is reported. Compound 3 was competitive and selective with pA2 values of 8.0 +/- 0.1 (rabbit) and 8.4 +/- 0.05 (rat) on smooth muscle preparations and 8.16 +/ 0.01 on human platelets. In vivo activity of 3 was demonstrated in a Konzett Rossler guinea pig model at 0.01 mg/kg p.o. PMID- 2973085 TI - Prostaglandin E2 and atriopeptin III oppose the contractile effect of angiotensin II in rat kidney mesangial cell cultures. AB - Effects of vasoconstrictory and of dilatory hormones were studied on the contractile activity of cultured rat kidney mesangial cells. By phase contrast microscopy, a rapid contraction was seen of most cells treated with angiotensin II (10(-6) - 10(-10) mol/L), which was sometimes followed by autonomous relaxation after 10 to 20 min. Prostaglandin E2 and atriopeptin III prevented the contractile effect of angiotensin II in a dose-dependent manner. Angiotensin II, but not atriopeptin III, stimulated prostaglandin E2 synthesis in mesangial cell cultures. PMID- 2973086 TI - Comparison of low dose nicotinamide versus benzamide, administered per os, as radiosensitizers in a C3H mammary carcinoma. AB - We have evaluated if any differences in tumor radiosensitization exist between the two adenosine diphosphate ribosyl transferase (ADPRT) inhibitors nicotinamide and benzamide at fractionated low doses. A significant radiosensitizing effect with nicotinamide at a 10 mg/kg per day dose was found in the tumor model used. We found, however, no radiosensitizing effect with benzamide given according to this schedule. PMID- 2973087 TI - A time to care. PMID- 2973088 TI - Oh, my back! PMID- 2973089 TI - Don't deny AIDS victims dental care. PMID- 2973090 TI - Characteristics of distension-induced release of immunoreactive atrial natriuretic peptide in isolated perfused rabbit atria. AB - Atrial pressure- or distension-induced release of atrial natriuretic peptide (ANP) has been considered as an important regulatory mechanism of ANP release in cardiac atria. A new technique to permit graded continuous atrial distension has been developed in an isolated perfused single rabbit atrium. Graded atrial distension was induced by changing the elevation of the outflow catheter tip. Intra-atrial volume expansion resulted in an increase in immunoreactive ANP release. The graded increase in atrial distension from 43.9 +/- 10.2 to 207.7 +/- 29.1 microliter resulted in 6.2-27.1-fold increases in volume-dependent immunoreactive ANP release. A rise in immunoreactive ANP release induced by increasing atrial distension did not occur in the state of atrial distension but occurred only after return to the reduced distension. However, in the case of atrial distension with pacing, an increase in immunoreactive ANP release was observed during atrial distension with pacing and after return to the basal level. The present study shows that the new technique is applicable to the study of the 'stretch-secretion coupling' mechanism of ANP release in vitro, and that the more important factor involved in the release of immunoreactive ANP induced by atrial distension may be the atrial reduction to basal level after distension rather than the stretch itself. PMID- 2973091 TI - [Testicular feminization syndrome. An echographic study of one family]. PMID- 2973093 TI - [Traumatic perforations of the tympanic membrane--its therapeutic resolution]. PMID- 2973092 TI - [Our experience in the medical treatment of chronic otorrhea]. PMID- 2973094 TI - [Our experience with ENT injuries in children]. PMID- 2973095 TI - [Phenytoin in the treatment of ear noises]. PMID- 2973096 TI - [Data on the physiology and physiopathology of lymph circulation of the nasal mucosa]. PMID- 2973098 TI - [Bilateral ligation of the external carotid in a case of post-tonsillectomy hemorrhage]. PMID- 2973097 TI - [Deafness as a sign of onset of middle ear cancer (a case presentation)]. PMID- 2973099 TI - [Phonation injury emergencies in an ENT outpatient clinic]. PMID- 2973100 TI - [Endoscopic indications and therapy at the Craiova District Hospital over a 5 year period (1980-1984)]. PMID- 2973101 TI - [Value of trephining puncture in the diagnosis and treatment of frontal sinusitis]. PMID- 2973102 TI - [Hormonal determination of the differentiation of striated skeletal muscle in urodele amphibians]. AB - In the urodelan amphibian Pleurodeles waltlii, spontaneous external metamorphosis was correlated with an increase in the serum level of thyroxine (T4). Within the same period, a change occurred in the myofibrillar ATPase profile of the dorsal skeletal muscle; fibres of larval type were gradually replaced by transitional fibres (type IIC), then by adult fibres of the types I, IIA, and IIB. Likewise, a myosin isoenzymic transition was observed. In larval animals, myosin electrophoresis revealed 3 bands corresponding with isoforms having identical heavy chains (MHC), but different light chains (MLC). In the course of metamorphosis, the 3 larval isomyosins were replaced by 3 isoforms having the adult type MHC and different motility. In a related neotenic species, Ambystoma mexicanum, no spontaneous anatomic metamorphosis occurred; at the time it should theoretically take place, the serum T4 level remained low. The ATPase profile was modified, but transitional fibres that replaced the initial larval types appeared to be persistent, and adult fiber types appeared only in a small amount. Myosin isoenzymic transition was also incomplete, larval isoforms were still distinguished in the neotenic adults. Similar persistence of larval characters was observed in adult Proteus anguinus, a perennibranch that never undergoes anatomical metamorphosis. Experimental hypothyroidian Pleurodeles waltlii displayed no external metamorphosis, only the larval fibre types and isomyosins were detected in those animals. External metamorphosis was induced in Ambystoma mexicanum by a triiodothyronine treatment. A complete myosin isoenzymic transition was observed in metamorphosed animals. These results tend to indicate that a moderate increase in the level of thyroid hormones is sufficient to determine the production of the adult type MHC molecules and the differentiation of the corresponding myofibrillar types in the skeletal dorsal muscle of amphibians, while a marked increase would be necessary for repressing the initial larval feature. PMID- 2973103 TI - Specific effects of low-frequency stimulation upon energy metabolism in tibialis anterior muscles of mouse, rat, guinea pig and rabbit. AB - Tibialis anterior (TA) muscles of four small mammals were subjected to chronic nerve stimulation for 28 days (10 Hz, 10 hours per day). Total cellular activities of phosphofructokinase (PFK), hexokinase (HK), citrate synthase (CS), 3-hydroxy-acyl-CoA dehydrogenase (HADH) and 3-hydroxybutyrate dehydrogenase (HBDH) were measured in the stimulated and unstimulated contralateral muscles. Normal TA muscles displayed ranges of oxidative and glycolytic capacities with rabbit TA showing the lowest and mouse TA the highest oxidative capacity. Chronic stimulation was almost without effect in mouse TA. In all other species, glycolytic capacity was decreased and reference enzymes of aerobic-oxidative pathways were increased. Rabbit TA displayed the highest increment in oxidative capacity with approximately three-fold increases in CS and HADH and eleven-fold increases in HBDH. Different responses were also observed for HK. In some cases, the extent of adaptation appeared to be independent of the initial enzyme activity levels, while in other cases it appeared to follow an order which corresponded to the size of the animals. Thus, there exist species-specific ranges of adaptation and adaptive alterations in one species may not necessarily reflect the adaptive response of another species. PMID- 2973104 TI - An experimental orthopedic treatment of the rat mandible using a functional appliance alters the fibre and myosin types in masticatory muscles. AB - Masticatory muscles mediate the action of functional orthopedic appliances on mandible growth. When young rats were treated for four weeks with a postural hyperpropulsor of the mandible, an appliance designed to increase condylar cartilage growth rate, the proportion of fast non-fatigable fibres in the lateral pterygoid muscle increased significantly. Concomitantly, the amount of slow myosin light chains increased in fibre extracts. This slow myosin originated from IIA fibres. By functional orthopedic treatment, the lateral pterygoid muscle was enriched in less fatigable fibres; the changes observed in the lateral pterygoid muscle were close to changes observed in other muscles after training. PMID- 2973106 TI - [Prevention of deviant behaviors under the current conditions of mental care in Romania]. PMID- 2973105 TI - Biochemical characteristics of mammalian diaphragms. AB - Selected biochemical characteristics of diaphragm muscle were compared among several orders of adult mammals (cattle, swine, rabbit, guinea pig, rat, and mouse) with known differences in resting breathing frequencies (f, range = 15 138). Diaphragms from smaller animals had significantly higher citrate synthase (CS) and phosphofructokinase (PFK) activities and substrate oxidation rates than larger animals. Ranges of activities for CS and PFK were 93-27 and 58-39 mumol.g 1.min-1, respectively; and 34-5 and 19-2 nmol.g-1.min-1 for [U-14C]glucose (GLU) and [1-14C]palmitate (PAL) oxidation, respectively. The percent of native fast myosin (FM) isoforms was significantly different among groups. Mouse diaphragm had the highest % FM (88.6%), whereas the lowest values (7.5%) were observed in cattle diaphragm. Myosin ATPase (M-ATPase, pH 9.8) activity was significantly lower in cattle (0.06 mumol.mg protein-1.min-1) and swine (0.38 mumol.mg protein 1.min-1) diaphragm than in other mammals (range of 1.14-0.67 mumol.mg protein 1.min-1). Correlation coefficients determined among means of measured biochemical parameters and established values of f indicated that CS activity and substrate oxidation rates were significantly correlated with f (r = 0.92, 0.92, 0.86 for CS, GLU, PAL, respectively) and the % FM increased with f. M-ATPase (pH 9.8) was significantly correlated with % FM (r = 0.85), whereas PFK and M-ATPase activities were not closely associated with f. It was concluded that f in mammals is significantly correlated with the biochemical parameters of aerobic capacity and is associated with the percent of FM isoforms in the diaphragm. PMID- 2973107 TI - [Forensic psychiatric expertise in 1962-1987]. PMID- 2973108 TI - [Forensic psychiatric expertise. A methodology letter]. PMID- 2973110 TI - [National Conference on Psychiatry and Legal Medicine. Predeal, 23-24 October 1987. Proceedings]. PMID- 2973109 TI - [The role of mourning and affective losses in the etiology of depressive states]. PMID- 2973111 TI - [The concept of responsibility in the function of mental capacity and its relationship to large groups of mental diseases arranged by the intensity of the disorders]. PMID- 2973112 TI - [Indolamines and sleep: 5-HT or 5-HTP?]. PMID- 2973113 TI - [Vertebral and spondylogenic syndromes]. PMID- 2973114 TI - [Spectrum analysis and conventional Doppler studies in the functional evaluation of aorto-iliac stenosis]. PMID- 2973115 TI - [Acute ischemia of the limbs. Etiology, treatment, prognosis]. PMID- 2973116 TI - Dominantly inherited spastic paraplegia and multifocal palmoplantar hyperkeratosis. AB - We have extended the study of a previously published kindred with spastic paraplegia and palmoplantar hyperkeratosis from 2 to 18 persons. Present evidence suggests that both neural and cutaneous manifestations are due to a single mutant gene, inherited as an autosomal dominant. Nerve conduction and EMG examination provide evidence for mild lower motor and primary sensory neuron (axon) involvement but these techniques are not useful for early detection. The disorder is a unique genetic disease which stands apart from other mostly recessively inherited varieties of neuroichthyosis. PMID- 2973117 TI - Modulation of immune responses by surface polysaccharides of Candida albicans. AB - Fungal polysaccharides, especially those from the cell wall of the nonpathogenic yeast Saccharomyces cerevisiae, have been investigated as modulators of immune function for many years. More recently, because surface-associated components are known to be elaborated and circulate during serious episodes of candidiasis, investigators have taken interest in the opportunistic fungus Candida albicans. This review emphasizes the immunomodulatory activities of mannan and cell wall derived glycoproteins from C. albicans. Mannan extracted with hot citrate buffer has been shown in an antibody-forming cell assay to be a heterogeneous mixture with components capable of enhancing or suppressing antibody responses when administered to mice at or near the time of immunization with type III pneumococcal polysaccharide (SSS-III), a T helper cell-independent antigen, or with sheep erythrocytes (SRBC), a T helper cell-dependent antigen. The components responsible for these opposing effects have been separated on the basis of size or charge by column chromatography. Two similar cell wall glycoproteins, removed from isolated cell walls by different procedures, induced enhancement only of the immune response to SSS-III and SRBC. The mechanism(s) by which these polysaccharides effect modulatory activity has not been elucidated. The enhancing property does not appear related to a direct mitogenic effect on lymphocytes or to stimulation of the production of B cell growth factors or interleukin 2. PMID- 2973118 TI - Influence of cryptococcal antigens on cell-mediated immunity. AB - Cryptococcosis is a frequently fatal disease caused by a yeast-like organism, Cryptococcus neoformans. Patients with systemic cyptococcosis generally have cryptococcal antigen in their body fluids; levels of antigen directly correlate with severity of the disease. With use of a murine model, it was documented that cell-mediated immunity is an important protective mechanism and that an epitope(s) on the mannoprotein fraction of the cryptococcal antigen present in a culture filtrate is recognized in cell-mediated immunity. Sera from mice infected with C. neoformans will induce suppressor cells that suppress cell-mediated immunity, as measured by the delayed-type hypersensitivity response, and the level of cryptococcal antigen in the serum directly correlates with the degree of suppression induced. There is no correlation with the amount of anticryptococcal antibody and the ability of the serum to induce suppression. Further, it was demonstrated that cryptococcal antigen, when injected into mice at concentrations similar to those found in human systemic cryptococcosis, induces a complex series of suppressor cells and factors that specifically suppress the delayed-type hypersensitivity response and protective immunity to cryptococci. PMID- 2973119 TI - Computed tomography as the primary radiological examination of lumbar spine. AB - A series of 235 patients examined by lumbar CT because of sciatica or other low back disorder was studied. The need of additional examinations and correlations to surgical findings were evaluated. Inadequate information was the cause of additional examination, mostly myelography in 20 patients (8.5%). It was concluded that lumbar CT is a suitable first examination of the lumbar spine in sciatica and low back pain. The indications to complementary myelography and its benefit are discussed. PMID- 2973120 TI - T cell subsets in patients with systemic lupus erythematosus. AB - T cell subsets were determined in blood of 16 patients with systemic lupus erythematosus (SLE), and 16 healthy individuals. SLE patients showed marked decrease in relative level of suppressor/cytotoxic T cells, slight decrease in helper/inducer T cells, and an increased helper/suppressor T cell ratio. The diminished level of suppressor/cytotoxic T cells was correlated with the clinical severity of disease. Cytofluorographic analysis of T cell subsets obtained from SLE patients did not show significant differences in homogeneity of the cell subpopulations as compared to healthy controls. PMID- 2973121 TI - Treatment of advanced prostatic carcinoma with a slow release depot LHRH analogue (Zoladex depot). AB - We treated 38 patients with newly diagnosed advanced prostatic cancer with monthly injections of a long acting depot preparation of a luteinizing hormone releasing hormone superagonist (Zoladex depot). After 6 months' treatment 10% of the patients had complete objective regression, 77% had partial objective regression, 3% stable disease and 10% had objective progression. The LHRH analogue does not have the metabolic side effects of oestrogens. Depot luteinizing hormone-releasing hormone analogue may well become a preferred alternative for patients with advanced prostatic cancer. PMID- 2973122 TI - Endocrinological effects of treatment with high doses of medroxyprogesterone acetate in patients with renal or prostatic adenocarcinoma. PMID- 2973123 TI - Compliance and subjective relief by corset treatment in chronic low back pain. AB - The aim of this questionnaire-based 12-month follow-up study of 113 patients was to elucidate patient compliance and subjective help achieved with a first, elastic or semirigid corset in chronic, idiopathic low back pain. Subjective help obtained from the corset was reported as excellent or good in 37% of the returned questionnaires. A total of 60% reported having worn the corset even during the preceding month. Low semirigid and elastic models were found to be better by the males and high semirigid ones by the females (p less than 0.001). Age, height, weight, body mass index, retirement or physical strenuousness of work showed no statistically significant correlation with the subjective relief gained from the corset. It is important that sufficient time be allocated to fitting the corset and that adequate information be provided about wearing the brace and about suitable trunk exercises. PMID- 2973124 TI - On the application of the WHO handicap classification in rehabilitation. AB - In order to study the usefulness and limitations of the WHO ICIDH Classification of handicap, a consecutive group of 76 stroke patients was studied at admission and discharge from a rehabilitation ward. All patients were graded with respect to orientation, physical independence, mobility and social integration. Most patients had at admission a combination of severe handicaps in several areas; this was less so at discharge. Nearly half of the patients did not change their degree of handicap during their stay at the rehabilitation ward. Improvement was not dependent on length of stay in the rehabilitation, age or sex. We have had difficulty in differentiating between handicap and disability as defined in the WHO system and we seriously question the utility of using three distinct categories of functional consequences of disease. We believe that the WHO system is useful to describe the type and pattern of disablement in a group of patients admitted to a rehabilitation ward and provides information on staffing demands, criteria for admittance and discharge. PMID- 2973126 TI - Fixed drug eruptions to maloprim. PMID- 2973125 TI - Microinjected DNA from the X chromosome affects sex determination in Caenorhabditis elegans. AB - The signal for sex determination in the nematode Caenorhabditis elegans is the ratio of the number of X chromosomes to the number of sets of autosomes (X/A ratio). By previous genetic tests, elements that feminized chromosomal males appeared to be widespread on the X chromosome, but the nature of these elements was not determined. In experiments to define a feminizing element molecularly, cloned sequences were added to chromosomally male embryos by microinjection into the mother. Three different X-chromosome clones, including part of an actin gene, part of a myosin heavy chain gene, and all of two myosin light chain genes, feminize chromosomal males. Both somatic and germline aspects of sex determination are affected. In contrast, about 40 kilobases of nematode autosomal DNA, phage lambda DNA, and plasmid pBR322 DNA do not affect sex determination. A feminizing region was localized to a maximum of 131 base pairs within an intron of the X-linked actin gene; a part of the gene that does not have this region is not feminizing. The results suggest that short, discrete elements found associated with many X-linked genes may act as signals for sex determination in C. elegans. PMID- 2973127 TI - [Current problems of medical health care for animal breeders]. PMID- 2973128 TI - [Organization of the extracurricular training of students in a department of social hygiene and public health organization]. PMID- 2973129 TI - [The role of Party organizations of the Don River area in the health education of workers during the first years of Soviet power]. PMID- 2973130 TI - [Principles in the building of a proctology service in the RSFSR]. PMID- 2973131 TI - A retrospective study of low-back pain in 38- to 64-year-old women. Frequency of occurrence and impact on medical services. AB - The occurrence of low-back pain (LBP) was investigated in a retrospective cross sectional study of a random sample of 1,760 38 to 64-year-old women. The lifetime incidence of LBP was 66% and the prevalence was 35%. Neither the lifetime incidence nor the prevalence rates were significantly associated with age. Inability to work because of LBP was found in 2.6% of the women in the 38- to 49 year age group and 5.9% among 50- to 64-year-old women (P less than 0.05). The utilization of medical services because of LBP was high, but only 1% of all investigated women had had a back operation. Forty-seven percent of the women in the prevalence group had experienced leg pain (sciatica), the frequency being significantly higher among the older women (P less than 0.01). Increased LBP when performing certain activities of daily living was common, and significantly more pronounced in the older age group. PMID- 2973132 TI - Compliance for low-back pain patients in the emergency department. A randomized trial. AB - A randomized trial was used to evaluate two forms of a Health Belief Model (HBM) intervention aimed at increasing compliance among low-back pain (LBP) patients visiting an Emergency Department (ED). Patients who received an HBM clinical intervention administered in the ED, HBM phone intervention made 1 to 2 days post ED visit, or both interventions were much more likely than control patients to both schedule and keep a follow-up referral appointment. Both the phone and clinical interventions were designed to increase patients' perceived susceptibility to complications of the LBP, seriousness of the complications, and benefits and costs of action through a follow-up referral appointment. Other factors predicting compliance included persistence of pain symptoms, previous treatment of LBP by a health care provider, and an unmet need for child care. PMID- 2973133 TI - Do nonorganic signs help to predict the return to activity of patients with low back pain? AB - The value of the nonorganic signs in predicting return to activity following an initial episode of low-back pain was determined. One hundred and twenty patients were assessed within 6 months of the onset of their first episode of disabling low-back pain and again at a mean of 15.3 months following injury. No correlation was found between the presence of nonorganic signs at initial assessment and either return to activity or resolution of the patient's symptoms. PMID- 2973134 TI - Early results of spinal fusion using variable spine plating system. AB - Seventy-seven consecutive patients underwent application of variable spine plating (VSP) spinal plates between August 1984 and October 1985. Sixty-four percent had previous procedures at the same level or levels operated. Operative indications were spinal stenosis, segmental instability, unstable spondylolisthesis, herniated disc with instability, pseudoarthrosis, unstable fracture, and failed surgery syndrome with evidence of one of the preceding. Overall results showed 30% excellent, 30% good, 34% fair, 6% poor. There were four deep wound infections and 19 patients with one or more broken screws. Screw alignment and the angular relationship of each screw to the spinal plate are considered important technical factors in minimizing screw failure. Vigorous distraction of the vertebrae using interpedicular screws is rarely indicated. Twenty-four patients required reoperation. We feel the procedure is relatively indicated in cases of moderate to severe instability, such as some cases of spondylolisthesis, failed surgery with marked segmental instability, the obese, deconditioned patient, or cases of spinal stenosis rendered very unstable by surgical decompression, and most strongly indicated in unstable lumbar and thoracolumbar fractures. PMID- 2973135 TI - Neuromuscular thermography of the lumbar spine with CT correlation. AB - Fifteen asymptomatic volunteers and 19 patients with current CT (GE 8800 CT/T) scans demonstrating either thecal sac contour distortion or nerve root displacement from disc rupture or spinal stenosis of the L4-L5 or L5-S1 levels judged to be at least moderate in severity underwent lumbar thermography. All patients were studied with an infrared telethermographic unit (AGA model 720M) employing the technical standards recommended by the Academy of Neuromuscular Thermography. The thermographic exams were interpreted independently and in a blind fashion by two radiologists. Of the 15 exams of asymptomatic patients, six were interpreted as positive and consistent with nerve fiber irritation. Of the 19 exams of patients with demonstrated CT abnormalities, all had positive thermograms for nerve fiber irritation (specificity 60%, sensitivity 100%). It is concluded that lumbar thermography is a sensitive examination for detecting those patients who will demonstrate lumbar spinal CT abnormalities and should play an important role in the diagnostic screening of low-back pain syndrome patients. PMID- 2973136 TI - The stress lumbar discogram. AB - Stress discography is that done during the time in which the subject's pain is present at a significant level of intensity. This prospective study examines the findings in 200 patients who were evaluated for low-back pain. A standardized injection technique was used and the contrast pattern within the disc was studied radiographically and with CT scanning. Pain response was recorded. Familiar low back pain was reproduced in all cases, and familiar leg pain in 46% of cases. Of 549 discs studied, 319 reproduced familiar pain, an average of 1.6 discs per patient. Pain reproduction occurred in 69% of the L5 discs, 62% of the L4 discs, and 25% of the L3 discs. Internal disc disruption was present in 302 discs, but extruded annular material was found in only 19. Stress discography is a useful diagnostic study in evaluating patients with low-back pain. PMID- 2973137 TI - Symptomatic clasp-knife deformity of the spinous processes. PMID- 2973138 TI - Report of the Disability Advisory Council: executive summary. PMID- 2973139 TI - Pneumoperitoneum complicating coronary bypass surgery: management without laparotomy. AB - Drainage of the mediastinum or thoracic cavity following bypass surgery is a routine procedure. A case is reported where pneumoperitoneum developed after the surgical procedure with vague abdominal symptoms accompanied by fever and leukocytosis. Because of the possibility of a rupture of an intra-abdominal organ laparoscopy was performed in this very ill patient and laparotomy avoided. Attention is drawn to this particular group of patients with the sequela of pneumoperitoneum in which laparoscopy can solve the diagnostic dilemma. PMID- 2973140 TI - The influence of hepatocellular function on NK and T cell tumoricidal activity. AB - Recent studies by this group have demonstrated that hepatocellular integrity is important in the preservation of host cellular immune function. This study evaluated the effect of experimental hepatocellular dysfunction (EHD) on host antineoplastic defense mechanisms. In nonspecific immune studies, we examined the effect of EHD on Wistar Furth (WF) natural killer (NK) cell cytotoxicity; in specific immune studies, we assessed WF C3H/HeJ lymphocytic responsiveness to both T cell mitogen and unmodified syngeneic fibrosarcoma. In concurrent studies, we evaluated the effect of EHD on interleukin-2 (IL-2) synthesis, an important NK and T cell trophic factor. WF rats and C3H/HeJ mice were assigned to three groups: EHD induced by bile duct ligation, sham, and normal control (NC). At day 21 serum bilirubin, WF NK cytotoxicity to YAC-1 tumor cells, WF and C3H/HeJ lymphocytic responsiveness to phytohemagglutinin (PHA) and syngeneic MCA fibrosarcoma (MCA-F), and WF T-helper IL-2 production were determined in respective groups. Serum total bilirubin was elevated in EHD rats and mice with respect to controls (p less than 0.01). Wistar Furth cytotoxicity to the YAC-1 tumor cells was depressed in EHD animals with respect to sham and NC groups at 12.5:1 (p less than 0.01), 25:1 (p less than 0.05), 50:1 (p less than 0.05), and 100:1 (p less than 0.05) effector/target cell ratios. WF T cell responsiveness to PHA was depressed in EHD with respect to controls (p less than 0.01). C3H/HeJ lymphoproliferative response to MCA-F tumor antigen was also depressed in EHD animals when compared with control groups with the addition of 12.5 X 10(3) (p less than 0.05) and 50 X 10(3) (p less than 0.05) MCA-F cells. These impairments in NK and T cell function in EHD could not be attributed to diminished IL-2 production (EHD vs sham and NC: 112,141 +/- 5232 vs 106,691 +/- 1419 and 120,759 +/- 3248 cpm, respectively). These results demonstrate that hepatocellular failure compromises NK and T cell tumoricidal function, an effect not resultant on diminished T helper IL-2 production. PMID- 2973141 TI - Distribution of dopamine D1 receptors in rat cortical areas, neostriatum, olfactory bulb and hippocampus in relation to endogenous dopamine contents. AB - The tritiated dopamine D1 antagonist SCH23390 was employed to determine the densities of D1 receptors in seven discrete and functionally identified cortical areas (cingulate, frontal, parietal, primary somatosensory, primary visual, retrosplenial and entorhinal-piriform) as well as in the neostriatum, hippocampus and olfactory bulbs. In addition, the tissue levels of the catecholamines NA, AD, DA, the indoleamine 5-HT and their main metabolites (MHPG, DOPAC, HVA, 3-MT, 5 HTP and 5-HIAA) were measured in the different regions by HPLC with electrochemical detection. The Scatchard analysis of saturation curves revealed the highest density of [3H]SCH23390 binding sites for the neostriatum, while the densities were 10-20 times lower for total cerebral cortex and hippocampus respectively. For the olfactory bulb and other cortical areas, D1 receptor densities were determined by equilibrium binding at a fixed radioligand concentration approaching saturation. The distribution of D1 receptors was heterogeneous with the greatest densities in entorhinal-piriform and cingulate cortices. The endogenous DA levels measured for all regions correlated with their metabolite (DOPAC, HVA and 3-MT) contents (r = 0.999; P less than 0.001). There was also a very good correlation (r = 0.981; P less than 0.001) between tissue DA and D1 receptor densities. This quantitative information reflects particular features of the organization of the DA systems and is discussed in relation to turnover and recently established aspects of the DA innervation. PMID- 2973142 TI - SKF 38393 alters the rate-dependent D2-mediated inhibition of nigrostriatal but not mesoaccumbens dopamine neurons. AB - Previous electrophysiological studies have failed to identify significant effects of the D1 dopamine (DA) agonist SKF 38393, either alone or in combination with the D2 agonist quinpirole (LY 171555), on the spontaneous firing rate of midbrain DA neurons. We have utilized extracellular single-unit recording techniques to examine whether SKF 38393 can alter D2-mediated inhibition of DA cell activity. Quinpirole-induced inhibition of the spontaneous activity of midbrain DA neurons was observed to be positively correlated with the basal firing rate of the neuron being examined (i.e., faster cells required higher doses to achieve 50% and maximal inhibition). Pretreatment with SKF 38393 (1.0 mg/kg, i.v.; 4 minutes) eliminated the rate dependency of quinpirole-induced inhibition of nigrostriatal but not mesoaccumbens DA neurons. This effect of SKF 38393 was blocked both by the D1 antagonist SCH 23390 and by hemitransections of the forebrain. In summary, SKF 38393 appears to exert Dl-specific, feedback pathway-dependent effects on the profile of responsiveness of nigrostriatal DA neurons to D2-mediated inhibition of cell firing rate. PMID- 2973143 TI - Norepinephrine uptake dependent upon apparent Mg++-ATPase activity and proton transport in storage vesicles in axoplasm. AB - A Ca++-dependent secretion of norepinephrine ([3H]NE) was evoked in adrenergic nerve endings in rat heart ventricle slices incubated in a modified Krebs-HCO3 medium containing choline Cl as the replacement for NaCl (Ch+-Ca++). Exogenous ATP inhibited secretion and lithium ion, a known inhibitor of NE uptake dependent upon Mg++-ATPase activity in vesicles (but not ouabain) prevented the response to ATP (Bogdanski, 1983,1986). It was suggested that vesicles attached to the axolemma recaptured [3H]NE from the extracellular fluid. This report indicates that other known inhibitors of uptake in isolated vesicles also inhibited the response to ATP in the attached vesicles. Included were two inhibitors of Mg++ ATPase activity, N-ethylmaleimide (NEM) and dicyclohexylcarbodiimide (DCCD), and the proton transporters 2,4-dinitrophenol (2,4-DNP 1.0 mM) and chlorpromazine (CPZ). Potassium ionophores (valinomycin with 2,4-DNP 0.1 mM, and nigericin) and a pH neutralizing reagent for vesicles (NH3 from ammonium sulfate in solution) were also effective. The uptake inhibitors, except 2,4-DNP, could also increase the rate of depletion of stored NE and its deamination in nonsecreting nerve endings incubated in Krebs-HCO3 (KRB) medium. Valinomycin by itself stimulated uptake in the presence of ATP. It is suggested that mechanisms of uptake and retention of NE in isolated vesicles (symposium (1982) Fed. Proc. 41:2742-2780) apply to the axoplasmic vesicles as well. Thus, the activity of Mg++-ATPase drives proton transport to establish the electrochemical gradients of H+, which drive the transport of NE. A lowering of the gradients can mobilize amines and evoke secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973144 TI - Biphasic modulation of evoked [3H]D-aspartate release by D-2 dopamine receptors in rat striatal slices. AB - It has been hypothesized that dopamine (DA) inhibits glutamate release from corticostriatal fibers via presynaptically located D-2 DA receptors although the evidence presented in the literature has not been conclusive. In the present experiments, the effect of D-2 receptor ligands on K+-stimulated tritium release from rat striatal slices preloaded with the nonmetabolizable glutamate analog [3H]D-aspartate ([3H]ASP was measured. The D-2 receptor antagonist S-sulpiride increased stimulated [3H]ASP release by 75% (EC50 value = 240 nM) and the biologically less-active isomer R-sulpiride, although equally effective, was tenfold less potent. The D-2 receptor agonists pergolide and (+)-4-propyl-9 hydroxynapthoxazine (+PHNO) inhibited [3H]ASP release at nM concentrations; however, this effect was small (20%). This low efficacy of the exogenous agonists was apparently due to competition by high concentrations of endogenous DA since the effect of pergolide was increased in rats whose striatal DA levels were decreased by 97%. These data support the hypothesis that D-2 DA receptors modulate [3H]ASP release in an inhibitory fashion. However, when the agonists were tested at lower concentrations, [3H]ASP release was increased significantly by 20% in control rats and 60% in DA-depleted rats. Both the facilitory and inhibitory effects of pergolide were blocked by 10 microM S-sulpiride, suggesting D-2 receptor mediation. In addition, the facilitory effect of pergolide was blocked by tetrodotoxin (TTX) and by the GABAA antagonist bicuculline, implying mediation of this D-2 effect by an inhibitory GABAergic interneuron. The inhibitory effect of pergolide was decreased by the muscarinic antagonist atropine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973145 TI - [3H]vesamicol binding in brain: autoradiographic distribution, pharmacology, and effects of cholinergic lesions. AB - An autoradiographic analysis of high-affinity binding sites for the vesicular acetylcholine transport blocker [3H]vesamicol (2-(4-phenylpiperidino) cyclohexanol; AH 5183) was conducted in rat brain. [3H]Vesamicol binding was displaced 52-99% by DPPN [( 2,3,4,8]-decahydro-3-(4-phenyl-1-piperidinyl)-2 napthalenol) (IC50 = 14 nM) and by ketanserin (500 nM), haloperidol (43 nM), and vesamicol analogs, but not by drugs selective for adenosine, adrenergic, amino acid, calcium channel, monoaminergic, opioid, PCP, sigma, or several other receptor classes. [3H]Vesamicol binding was most concentrated in the interpeduncular nucleus and fifth and seventh cranial nerve nuclei. Moderate binding was found in the lateral caudate-putamen, medial nucleus accumbens, olfactory tubercle, vertical and horizontal diagonal bands of Broca, and basolateral amygdala. The distribution of [3H]vesamicol binding was similar to distributions of acetylcholine (r = 0.88), acetylcholine esterase (r = 0.97), choline acetyltransferase (ChAT) (r = 0.97), and [3H]hemicholinium-3 binding sites (r = 0.95-0.99). Lower correlations were obtained between [3H]vesamicol and muscarinic receptor densities (r = 0.50-0.70). Few exceptions to the match between binding and cholinergic neuronal markers were found, e.g., the molecular layer of the cerebellum and the thalamus. Lesions of cholinergic neuronal projections to the neocortex or hippocampus reduced [3H]vesamicol binding in each of these regions, but to a lesser extent than reductions in ChAT. [3H]Vesamicol binding sites appear to be anatomically associated with brain cholinergic neurons, a locus that is consistent with the control by this site of vesicular acetylcholine uptake. PMID- 2973146 TI - Selective cortical infarction reduces [3H]sulpiride binding in rat caudate putamen: autoradiographic evidence for presynaptic D2 receptors on corticostriate terminals. AB - Although the existence of presynaptic D2 dopamine receptors on corticostriate terminals has been supported by numerous receptor-binding studies, recent autoradiographic data has failed to demonstrate loss of striatal D2 receptors following cortical lesions. In the present study, Long-Evans rats were subjected to unilateral middle cerebral artery (MCA) infarction in order to produce reproducible lesions of the neocortex without damaging subcortical structures. Animals were sacrificed 2 and 4 wk following lesion and brains were prepared for receptor autoradiography. D2 receptors were studied using the selective ligand [3H]sulpiride, while D1 dopamine receptors were examined using [3H]SCH 23390. Sodium-dependent, high-affinity choline uptake sites were labeled with [3H]hemicholinium-3, thereby providing a quantitative measure of cholinergic neuronal integrity. Unilateral cortical infarction resulted in approximately a 20% reduction in [3H]sulpiride binding in several discrete regions of the ipsilateral caudate-putamen (CPu), but not in the nucleus accumbens. D2 receptor binding was also reduced significantly in some areas of the contralateral CPu when compared with [3H]sulpiride binding in sham-operated, control animals. In contrast, D1 receptors (as identified by [3H]SCH 23390 and high-affinity choline uptake sites (labeled with [3H]-HC-3) were not affected by the cortical lesion. The results provide autoradiographic confirmation of the existence of presynaptic D2 receptors on corticostriate terminals. PMID- 2973148 TI - [Effects of 2 doses of lysine acetylsalicylate on platelet aggregation, beta thromboglobulin and platelet factor 4 in aged atherosclerotic patients]. PMID- 2973147 TI - Organization of dopamine D1 and D2 receptors in human striatum: receptor autoradiographic studies in Huntington's disease and schizophrenia. AB - The technique of quantitative autoradiography was used to examine the effects of Huntington's disease (HD) and schizophrenia on the organization of striatal dopamine (DA) D1 and D2 receptors. Whereas the striatum of HD cases showed a reduction in the density of D1 ([3H]SCH 23390) and D2 ([3H]spiroperidol) receptors, the patterning of D2 receptor loss did not match that of the D1 receptor loss. The HD loss of D1 D1 receptors (65%) is far greater than the loss of D2 receptors (28%). Whereas there was a dorsal-ventral gradient of effect on both receptor subtypes, the effects of HD on D2 receptors in the ventral putamen (PUT) and nucleus accumben septi (NAS) were minimal. Similarly, muscarinic M1 and M2 receptors demonstrate different patterns of alteration in HD. The M2 subtype, labeled with [3H]N-methylscopolamine (in the presence of excess pirenzepine to occlude M1 sites), was depleted far more than the M1 receptor subtype, labeled with [3H]pirenzepine. Although the effects of HD on [3H]mazindol labeling of DA terminals were more heterogeneous, there appeared to be a relative preservation of this afferent input to the striatum of the HD cases. In the schizophrenic cases, our autoradiographic studies confirm previous reports of an elevation of D2 receptor density in the striata of many schizophrenics. This increase was evident even though two of the three cases were known to have not been treated with neuroleptics, and the third case may also have been drug naive. However, the increase was far greater in the NAS (164%) and ventral PUT (173%) than more dorsally in the striatum (68%). The density of D1 receptors and DA terminals labeled with [3H]mazindol in the striatum of schizophrenics was not significantly different from that of control cases. Thus in both HD and schizophrenia, the ratio of D2/D1 receptors is altered in favor of the D2 population, particularly in the NAS. PMID- 2973149 TI - Mechanism of rabbit platelet agglutination induced by acidic mucopolysaccharide extracted from Stichopus japonicus Selenka. AB - The effects of acidic mucopolysaccharide extracted from sea cucumber (Stichopus japonicus Selenka) (SJAMP) on rabbit platelets were studied. Using citrated platelet-rich plasma (PRP), washed platelets, and formaldehyde fixed platelets from 10 New Zealand white rabbits, we investigated the effects of platelet inhibitors and various plasma and its fractions on SJAMP-induced agglutination. It was found that the tracing of platelet agglutination induced by SJAMP showed a single phase without a lag period. The lowest concentration of SJAMP required for the agglutination of rabbit platelets was approximately 2 micrograms/ml, and the magnitude of agglutination induced by SJAMP was concentration dependent. In 8 out of 10 rabbits, the platelets in PRP were agglutinated by 10 micrograms/ml of SJAMP. Platelet inhibitors, such as aspirin, indomethacin, apyrase, antimycin, 2 deoxy-D-glucose and EDTA did not inhibit the agglutination induced by SJAMP. Washed rabbit platelets were not agglutinated by SJAMP even though the concentration of SJAMP was raised up to 50 micrograms/ml. When rabbit plasma, serum, or 50-60% ammonium sulfate saturated plasma fraction was added to the reaction mixture, agglutination of washed platelets by SJAMP was recovered completely. But human plasma or fibrinogen did not have any effect on the reactivity of washed rabbit platelets to SJAMP. From these data we conclude that the SJAMP-induced rabbit platelet agglutination is independent of energy metabolism but requires plasma cofactor(s) other than fibrinogen. The plasma cofactor is present in 50-60% ammonium sulfate saturated plasma fraction. PMID- 2973151 TI - Mafenide (Sulfamylon) inhibits plasmin fibrinolytic activity. AB - Inflammatory fibrinolysis by plasmin or phagocyte proteases is a major cause of skin graft failure on burn wounds where the primary adherent attachment of the skin grafts is due to the glue-like action of fibrin. We investigated the potential of mafenide acetate solution, an experimental topical antimicrobial used in treating grafted burn wounds, to modify plasmin fibrinolytic activity in vitro and, thus, its potential to alter or modify the integrity of the fibrin glue critical for skin graft viability. Immobilized 125I-fibrin monolayers were used to assay fibrinolytic activity from plasmin or from plasma activated by streptokinase or urokinase and modified by the presence of mafenide or epsilon aminocaproic acid (EACA). While streptokinase-activated plasma lysed 52.7 +/- 3.9% of the 125I-fibrin, this plasmin activity was more than 80% inhibitable by EACA. Mafenide acetate had no intrinsic fibrinolytic activity (1.5 +/- 0.3%) nor activated plasma fibrinolytic potential (2.4 +/- 0.5%), but produced significant and dose-related reduction in fibrinolytic activity (p less than 0.001). Other sulfonamide analogues lacking a para-methylamino reactive group had 10-100 fold less antifibrinolytic potency while lysine, like mafenide, able to compete for plasmin binding sites, could potently block fibrinolysis. Mafenide did not qualitatively alter activation of plasminogen or affect generation of complexes with alpha 2 antiplasmin complexes. Adding mafenide only minutes following streptokinase-activated plasma or plasmin with the fibrin substrate reduced antifibrinolytic activity, supporting the conclusion that mafenide, like EACA, can modulate the interaction between fibrin and the plasmin reactive sites and thus prevent close plasmin/fibrin apposition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973150 TI - Aggregation of human platelets by acidic mucopolysaccharide extracted from Stichopus japonicus Selenka. AB - The acidic mucopolysaccharide extracted from sea cucumber (Stichopus japonicus Selenka) (SJAMP) has been shown to cause platelets to aggregate. Using citrated platelet-rich plasma (PRP), washed platelets and formaldehyde-fixed platelets from humans, we investigated the effects of platelet inhibitors and various plasmas and their fractions on SJAMP-induced platelet aggregation. It was found that the lowest concentration of SJAMP required for the aggregation of human platelets was about 0.4 micrograms/ml and the magnitude of aggregation induced by SJAMP was concentration dependent. The platelets were aggregated by SJAMP at 10 micrograms/ml in 25 out of 28 (89%) normal subjects tested. Platelet inhibitors such as PGE1, aspirin, indomethacin, apyrase, antimycin, 2-deoxy-D-glucose and EDTA inhibited by 70 to 100% the aggregation induced by SJAMP. Washed platelets alone were not aggregated by SJAMP. In the presence of fibrinogen, washed platelets were aggregated by SJAMP but formaldehyde-fixed platelets were not. These data indicate that the SJAMP-induced human platelet aggregation requires extracellular calcium, fibrinogen, and energy metabolism. The second phase of aggregation is dependent upon the release of ADP, and cyclooxygenase pathway. PMID- 2973152 TI - Serial determinations of PF4 and beta TG: comparisons between multiple venipunctures vs a catheter infusion system. AB - Platelet factor 4 (PF4) and beta thromboglobulin (beta TG) are platelet-specific proteins which are released upon platelet aggregation and which can be accurately measured by radioimmunoassay. We devised a catheter-infusion system that enables serial determinations of these proteins. In 20 subjects (10 healthy volunteers and 10 patients with stable coronary artery disease), we compared samples collected by individual venipunctures with those simultaneously obtained by means of a simple catheter-infusion system. At least 5 samples were obtained over a period of time which was as long as 60 min, and at least 30 min. Subjects with stable coronary artery disease were selected so that they would be expected to have stable and normal PF4 and beta TG levels. Thus, elevations of either PF4 or beta TG would represent artifacts secondary to sampling technique. Analysis of the results demonstrated that the catheter-infusion system was equivalent to individual venipunctures for determination of PF4 and beta TG. 16.8% of samples obtained via the catheter and 17.2% of those obtained by individual venipunctures were spuriously elevated. A second series of studies were performed to refine the technique further by examining the impact of infusion rate and the addition of citrate phosphate dextrose (CPD) to the infusate. Ten additional subjects had catheter systems utilized in both arms simultaneously. The addition of CPD resulted in significantly less abnormal values at slower infusion rates (1 and 2.5 cc/min). At 5 cc/min D5/w or saline alone are suitable. These investigations confirm that this simple catheter system is equivalent to individual venipunctures for determination of PF4 and beta TG while avoiding patient discomfort.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973153 TI - Familial type I protein S deficiency associated with severe venous thrombosis--a study of five cases. PMID- 2973154 TI - A monoclonal antibody to the human platelet glycoprotein IIb/IIIa complex induces platelet activation. AB - The platelet glycoprotein (GP) IIb/IIIa complex functions as the receptor for fibrinogen on activated platelets. The effects of two anti-GPIIb/IIIa monoclonal antibodies on platelet function were studied. These antibodies, 6C9 and C17, recognized different epitopes, which were exclusively present on the undissociated GPIIb/IIIa complex. Whereas C17 inhibited the binding of fibrinogen to platelets and platelet aggregation induced by adenosine diphosphate (ADP) or collagen, 6C9 caused irreversible aggregation of platelets, both in the presence and absence of extracellular fibrinogen. When incubated with unstirred (non aggregating) platelets, 6C9 induced release of alpha and dense granule constituents as well as binding of 125I-fibrinogen to platelets. The latter was evidently mediated in part by platelet-derived ADP, since it was inhibited to a large extent by apyrase, the ADP-hydrolyzing enzyme. F(ab')2 fragments of 6C9 did not induce platelet-release reactions but caused (slow) aggregation of platelets in the presence of extracellular fibrinogen. These results indicate that binding of an antibody to a specific site on the platelet GPIIb/IIIa complex may cause fibrinogen-mediated aggregation. The Fc part of the platelet-bound antibody appears to be involved in the induction of platelet release. PMID- 2973156 TI - Differential autolysis of human plasmin at various pH levels. AB - Denaturation of human plasmin in solutions of various pH levels was studied. The denaturation and loss of catalytic activity of plasmin in solutions of between pH 3.5 and 10.5 is a second-order kinetics. In alkaline solutions of pH levels greater than 11.5, plasmin undergoes a first-order denaturation. The second-order denaturation of plasmin is mainly due to the autolytic reactions between plasmin molecules. Two autolytic processes of human plasmin in aqueous solution were observed. In a slightly acidic solution (pH 6.5) the light (B) chain was found to be cleaved faster than the heavy (A) chain of plasmin. On the other hand, the heavy (A) chain was cleaved in an alkaline solution of pH near 11.0. A cleaved heavy (A) chain of molecular weight 58,000 was observed. Both the heavy (A) chain and the light (B) chain were found to be cleaved at pH levels between 6.5 and 11.0. The loss of the esterase activity of plasmin samples in the autolytic process is in parallel with the decline of intact light (B) chain. The autolytic cleavage of the heavy (A) chain led to the formation of a new type of catalytically active plasmin. PMID- 2973155 TI - Neutralization of thrombin by antithrombin III in the presence of cultured human fibroblasts. AB - Recent evidence suggests that heparan sulfate on endothelial cell surfaces acts as a catalyst for the neutralization of thrombin by antithrombin III (AT III). Fibroblasts also produce heparan sulfate which is present on the cell surface and secreted into the extracellular matrix. We evaluated the ability of cultured human fibroblasts to catalyze the interaction between thrombin and AT III and found that heparan sulfate produced by post-confluent fibroblasts was anticoagulantly active. Furthermore, after initial binding of thrombin to cells, thrombin-heparan sulfate appeared in the fluid phase above the cells; this thrombin could be rapidly neutralized by AT III independent of the further presence of cells. These results indicate that fibroblasts do produce an anticoagulantly active species of heparan sulfate and that the normal interaction between AT III and thrombin may be driven by initial release of heparan sulfate from the cell surface by thrombin followed by AT III interaction with the soluble thrombin-heparan sulfate complex. PMID- 2973157 TI - Increased plasma concentrations of complement modulating proteins (C1 inhibitor, C4-binding protein, factor H and factor I) in psoriasis. AB - By using single radial immunodiffusion we measured the plasma levels of four complement modulating proteins, i.e., C1 inhibitor, C4-binding protein, factor H and factor I in 19 psoriatic patients in comparison with those of healthy controls. Except for C1 inhibitor which was only marginally elevated, they were found to be significantly increased in psoriatic patients. When psoriatic patients were classified into subgroups based on the clinical severity, the levels of factor H and those of factor I showed a close positive correlation with the activity and extent of the skin lesions, whereas such clear relationship could not be found with C1 inactivator or with C4-binding protein. These results offer additional support for the hypothesis that the complement system is involved in psoriasis. PMID- 2973158 TI - [Comparative analysis of the degree of dental caries susceptibility of different ethnic groups]. PMID- 2973159 TI - [The rationale for primary prevention of dental caries and experience with its application to school children in the Arkhangelsk Oblast]. PMID- 2973161 TI - Interobserver agreement for the assessment of handicap in stroke patients. PMID- 2973162 TI - Effect of human atrial natriuretic peptide on postischemic acute renal failure in conscious dogs. PMID- 2973160 TI - In vitro and in vivo studies demonstrating potent and selective estrogen inhibition with the nonsteroidal aromatase inhibitor CGS 16949A. AB - CGS 16949A inhibited the conversion of [4-14C]androstenedione (A) to [4 14C]estrone by human placental microsomes in a competitive manner (Ki = 1.6 nM). Aminoglutethimide, also a competitive inhibitor, had a Ki = 0.7 microM in this assay system. The Km for the aromatization of A was 0.11 microM. Using ovarian microsomes from immature rats primed with pregnant mare's serum gonadotrophin and using [4-14C]testosterone conversion to [4-14C]estradiol as a measure of aromatase activity, the Km was 42 nM. At a substrate concentration 3-fold the Km, CGS 16949A was 180 times more potent as an inhibitor than aminoglutethimide, exhibiting half-maximal inhibition at 1.7 nM as compared to 0.3 microM. In vivo CGS 16949A lowered ovarian estrogen synthesis by gonadotropin-primed, androstenedione treated, immature rats by 90% at a dose of 260 micrograms/kg (PO). A dose of 100 mg/kg of aminoglutethimide was needed to produce this same effect. CGS 16949A at a dose of 4 mg/kg (PO) induced uterine atrophy (aromatase inhibition) without inducing adrenal hypertrophy - indicating a lack of inhibition of corticosterone secretion, while aminoglutethimide at 40 mg/kg (PO) induced adrenal hypertrophy without inducing uterine atrophy. CGS 16949A was neither androgenic nor estrogenic in rats using standard bioassays. The data suggest that CGS 16949A may serve as a potent and selective agent for modulating estrogen-dependent functions. PMID- 2973163 TI - Beneficial effect of atrial natriuretic factor on ischemically injured kidneys in rats. PMID- 2973165 TI - Hyperkeratosis palmoplantaris (Papillon-Lefevre syndrome). A case report. AB - A 16-year-old female presented with a severe chronic periodontitis and with associated hyperkeratotic lesions of her palms and soles. Investigations revealed it as a case of hyperkeratosis palmoplantaris (Papillon-Lefevre syndrome), which is a rare form of an ectodermal dysplasia. Previously it was speculated that this disease entity is not all that uncommon in Nigeria especially amongst the rural population. However, recent investigations revealed that this is not correct. PMID- 2973164 TI - Beneficial effects of maximal hydration on early renal function after ischemia is mediated by atrial natriuretic factor. PMID- 2973166 TI - [Unusual cases of the inheritance of Down's syndrome]. AB - Three unusual cases of Down's syndrome inheritance are presented which permit supposing that microstructural chromosome aberrations in parents may induce not only a structural imbalance of chromosomal material in progeny, but also a numerical imbalance. PMID- 2973167 TI - [Interaction of the enzymes of cellular energy support with the F-actin of the thin filaments of muscle fiber. I. The binding of lactate dehydrogenase with F actin induces changes in the structural state of the components of the complex]. AB - A study was made of changes in F-actin conformation occurring in a myosin-free single ghost fibre induced by the binding of glycolytic enzyme lactate dehydrogenase (LDG) to F-actin. The formation of the complex between LDG and F actin induces changes in the parameters of the intrinsic (tryptophan) and extrinsic (rodominil--phalloin) polarized fluorescence of F-actin of the ghost muscle fibre. It is found that LDG stimulates Mg2+-ATPase of actomyosin in solution. It is assumed that the coupling of energy-providing mechanism with that of muscle contraction may be accomplished through the conformation changes in F actin. PMID- 2973168 TI - Combining imaging and vascularity assessment of breast lesions. AB - Two separate series of patients were investigated to determine the improvement in diagnostic accuracy obtained by combining the results of ultrasonic imaging and vascularity assessment for the detection and diagnosis of breast disease. In the first series, ninety-five patients were examined with a hand-held Doppler probe to verify that abnormal flow could be detected in the presence of breast disease. To determine the clinical efficacy of combining the two examination procedures, a second series of patients was examined with the Doppler transducers incorporated into the water-bath scanner so that the sample volume could be accurately positioned into specific regions previously imaged. In the one hundred patients examined with both procedures, there was an improvement in the accuracy of diagnosing breast disease due to the reduction in the number of false positive and negative reports compared to the accuracy of the imaging examination alone. PMID- 2973170 TI - Xanthogranulomatous pyelonephritis: comparison of extent using computed tomography and magnetic resonance imaging in one case. AB - In a case of diffuse xanthogranulomatous pyelonephritis, computed tomography (CT) and magnetic resonance (MR) were used. The MR proved to be more precise in the preoperative evaluation of inflammatory extension to the spleen and into the abdominal wall. The CT was more accurate in excluding spread to the colon. PMID- 2973169 TI - Evaluation of total versus partial androgen blockade in the treatment of advanced prostatic cancer. AB - In order to evaluate the proposed benefit of complete androgen blockade in the treatment of patients with advanced prostatic cancer, we initiated a multicenter prospective and randomized study. At the time of this report 99 patients with newly diagnosed, previously untreated prostatic cancer were randomly distributed to one of the following treatments: group I, orchiectomy plus antiandrogen Flutamide; group II, depot LH-RH analog Zoladex plus Flutamide; group III, orchiectomy alone, and group IV, Zoladex alone. Our preliminary data fail to demonstrate a superiority of total androgen blockade over partial androgen blockade in the treatment of patients with advanced cancer of the prostate. PMID- 2973172 TI - [Role of ambulatory care in the dynamics of the incidence of chronic suppurative otitis media in adolescents]. PMID- 2973171 TI - Comparison of Megace, Stilphostrol, Megace plus DES, or streptozotocin in metastatic prostatic cancer in patients with hormonal failure and prior radiotherapy. AB - The National Prostatic Cancer Project from 1982 to 1985 evaluated several treatments for metastatic prostatic cancer patients who had a history of prior radiotherapy and were refractory to hormone manipulation. The treatments studied were megestrol acetate (Megace), Megace plus diethylstilbestrol (DES), diethylstilbestrol diphosphate (Stilphostrol), and streptozotocin. While the four treatment arms did not differ significantly with respect to survival, there was a small but significant difference in progression-free survival among the treatment groups. These patients are difficult to treat and have many secondary problems, and perhaps future studies of current and new agents should focus more on subjective and other secondary benefits for them. PMID- 2973174 TI - Immune complex-mediated glomerulopathy in Marek's disease. AB - Chickens infected with Marek's disease (MD) virus developed immune complex (IC) mediated glomerulopathy. Fluorescent antibody staining technique using antichicken globulin and antichicken complement was used to demonstrate IC in the kidney glomeruli. During the initial stages of MDV infection, IC deposits were seen on the glomerular basement membrane, but subsequently the entire glomerulus was involved. Mesangial cells also had IC deposits. Chicken complement was demonstrated in the glomeruli which had IC deposits. The number of glomeruli with IC deposition was higher in tumor-bearing birds than in non-tumor-bearing birds. Histologically, kidney lesion were characterized by thickening of basement membrane and proliferation of mesangial cells. It is suggested that IC-mediated glomerulopathy might be one of the major causes of death in MD. PMID- 2973176 TI - Partial FI gene-independence of lambda-21 hybrid phages specifying chimeric terminases. AB - The role of the FI gene in the life cycles of a series of lambda-21 hybrid phages that produce chimeric lambda-21 terminases has been examined. An isogenic series of FI+ and FI- derivatives of the hybrids was constructed, and the growth properties of the phages were examined. It was found that three of the four hybrids (hybrids 51, 67, and 33) are able to form plaques and produce a small burst in the absence of the FI gene product (gpFI), but each of the three phages is much healthier in the presence of gpFI. It is concluded that each of the three chimeric terminases is dependent on gpFI. The ability of the FI- hybrids to grow better than lambda FI- is postulated to be due to a minor qualitative or quantitative difference between the chimeric terminases and lambda terminase. The fourth hybrid (54), known from earlier work to produce an infirm terminase, is more dependent on gpFI than the other hybrids and lambda itself. PMID- 2973177 TI - [Results of the census of cancer patients receiving treatment in Leningrad hospitals in 1982]. PMID- 2973173 TI - Enumeration of lymphocyte subsets in peripheral blood of rhesus monkeys prior to and following monocyte removal using a modified indirect avidin-biotin immunoperoxidase technique. AB - A modified avidin-biotin immunoperoxidase technique has been used to enumerate B lymphocytes, T inducer/helper (TH) and T cytotoxic/suppressor (TS) cells in the peripheral blood of normal and immunosuppressed rhesus monkeys (Macaca mulatta) prior to and following adherent-cell depletion. The levels of each of the B, TH and TS cells detected in the normal monkeys using monoclonal antibodies which recognized specific surface antigens on human lymphocytes were comparable to the levels reported in human peripheral blood using direct immunofluorescence, immunoperoxidase or flow cytometry techniques. Adherent cell depletion did not result in a significant loss of any of the lymphocyte subpopulations examined. The technique is reproducible and sensitive in detecting differences between normal and immunosuppressed animals, and would prove to be useful in studies pertaining to chemical and drug immunomodulation. PMID- 2973175 TI - Preliminary observations on the effect of Dictyocaulus filaria on the blood clotting time of sheep. AB - The whole blood clotting time progressively increased from the second to the eighth week in lambs receiving a primary infection of Dictyocaulus filaria larvae. However, in vaccinated lambs it remained unaffected. Levamisole hydrochloride was ineffective when given 4 days after infection but restored the blood clotting time to near normal soon after treatment when it was given 30 days after infection. PMID- 2973178 TI - [Physician errors in breast cancer]. AB - An analysis of the data on 530 patients with breast cancer showed that errors made in diagnosis, treatment strategies and organizational arrangements had a significant adverse influence on survival in 212 cases. PMID- 2973179 TI - [Characteristics of the surgical stage in the combined treatment of poorly differentiated cancer of the corpus uteri]. AB - Ordinary extirpation of the uterus with appendages and on-indication lymphadenectomy followed by complex radiotherapy were given to 30 patients with anaplastic endometrial carcinoma at the first stage of treatment. Tumor advancement took place in 40 cases (17.4%) of extensive involvement and deep invasion of the organ. Large-fraction intracavitary gamma therapy from high energy 60Co sources and progestin treatment were carried out preoperatively in another 30 cases who subsequently underwent similar surgery. Postoperative treatment included hormones only. No untoward side-effects of intracavitary gamma or radiotherapy followed; nor were there surgical complications. Dissemination occurred in 2 (6.6%) cases of primary extrauterine extension of tumor. In group 2, patients' stay at hospital decreased by half. PMID- 2973180 TI - [Acid mucopolysaccharide content of the blood in patients with diffuse toxic goiter and ophthalmopathy]. PMID- 2973182 TI - Metabolism of 14C-thymoxamine in rat and man. AB - 1. After oral administration of 14C-thymoxamine to rat and man the total 14C excreted in urine and faeces was determined. 2. Six metabolites were isolated from the excret of man and rat by chemical extraction and identified by g.l.c. mass spectral analyses. 3. Two other metabolites, highly polar and resistant to enzymic hydrolysis, were isolated by extraction on XAD2 resin and h.p.l.c. analysis. These two metabolites were identified by n.m.r. and by mass spectrometry in the fast atomic bombardment mode. 4. These two major metabolites of thymoxamine in man and rat have been identified and characterized as the sulphate conjugates of desacetyl-thymoxamine and N-monodesmethyl-desacetyl thymoxamine. PMID- 2973181 TI - [Our therapy concept in nonresectable liver metastases]. AB - Cytotoxic chemotherapy was performed in a total of 18 patients (12 men, 6 women): 5 patients with colonic carcinoma and 1 patient with unknown primary lesion received 5 x 1000 mg 5-Fluorouracil (5-FU) at 4 week interval. The 5 following patients primarily suffering from colonic carcinoma were treated with 0.5 mg/kg BW FUDR continuously at 2 week interval. 5 further patients with colonic carcinoma sequential received Mitomycin C (8 mg/m2) and 4 x 1000 mg 5-FU. 2 patients with breast cancer were treated with 500 mg/m2 Cyclophosphamide, the same amount of 5-FU and 40 mg/m2 Methotrexate every 4 weeks. Chemotherapy was well tolerated by all patients. A clinically significant response, however, was seen in only 2 patients with breast cancer. In 8 patients a liver transplantat was performed, which was followed in 3 cases by ultra-high dose Cyclosphosphamide, lethal total body irradiation and autologous bone marrow transplantation. 1 further patient received polychemotherapy. At the time of this analysis only 3 patients were still alive at 61, 30 and 26 months with only 1 perioperative death. All 3 had meanwhile developed recurrent or metastatic disease. Because of these sobering results, liver transplantation for the treatment of non-resectable liver metastases has been abandoned, and regional chemotherapy is now only applied in patients with liver metastases from breast cancer and after resection of metastases in an adjuvant setting. PMID- 2973184 TI - [New lipid biochemical aspects in the pathogenesis of a follicular keratinization disorder in acne vulgaris]. AB - Follicular hyperkeratinization of the epithelium of the acroinfundibulum of sebaceous follicles is one of the primary events in the pathogenesis of acne vulgaris. Oral treatment with 13-cis-retinoic acid can reduce these hyperkeratoses. In order to determine whether follicular hyperkeratinization is related to disturbances of epidermal follicular lipids, we analyzed the lipids of initial comedones from 10 patients with nodulocystic acne before and after a 6th weeks oral therapy with 13-cis-retinoic acid (0.7 mg/kg body weight). The treatment with retinoid resulted in a significant increase of epidermal lipids (free sterols: +34%; ceramides: +19%, whereas the lipids of sebaceous origin decreased (glycerides: -36%). The mass ratio of free sterols to cholesterol sulfate increased by 86% compared to pre-treatment levels. These findings support the hypothesis that local follicular deficiencies of epidermal lipids due to increased sebum secretion might induce abnormal follicular keratinization. PMID- 2973183 TI - Influence of medium composition on 7-alkoxycoumarin O-dealkylase activities of rat hepatocytes in primary maintenance culture. AB - 1. The activities of 7-methoxycoumarin (7-MCOD), 7-ethoxycoumarin (7-ECOD) and 7 propoxycoumarin (7-PCOD) O-dealkylases decreased by 75-90% during culture of rat hepatocytes for 72 h. 2. The addition of dexamethasone (D) produced a stabilization or modest enhancement of 7-ECOD and 7-PCOD activities depending on the medium used; D was without effect on 7-MCOD activity. 3. The addition of nicotinamide (N) produced some stabilization of 7-ECOD and 7-PCOD activities but not of 7-MCOD activity. 4. The addition of D + N was associated with large increases in 7-ECOD and 7-PCOD activities, again with little effect on 7-MCOD activity. 5. Ethoxyresorufin O-dealkylase activity correlated with 7-ECOD/7-PCOD activities, whereas pentoxyresorufin O-dealkylase activity correlated with 7-MCOD activity. 6. It is concluded that D exerts a selective effect on certain forms of cytochrome P-450, but that more than one mechanism is probably implicated in this effect. 7. The magnitudes of induction of 7-ECOD activity by phenobarbitone and beta-naphthoflavone were blunted when cells were treated in culture when compared to treatment in vivo; the presence of D in the culture medium did not modify this phenomenon. PMID- 2973185 TI - [Surgical therapy of the acne tetrad]. AB - On the basis of a 3 years experience with surgical treatment of acne tetrad (acne conglobata, apocrine acne) in 22 patients, we can draw the following conclusions: 1. Radical surgical excision yields better long-term results than local incision. Recurrences are mostly the result of non-radical surgery. 2. In cases of limited acne--especially if it is localized either in the axilla or the groin--primary wound closure may be successfully attained. 3. Gentamycin chains may support the primary wound healing in defect closures of infected areas. 4. In the axilla, we suggest wound closure by myocutaneous island flaps as the best procedure after extensive excision. 5. Specific antibiotic protection, perioperatively, according to previous culture and sensitivity testing can guarantee a postoperative course free from infection. 6. Open wound therapy with secondary healing of the soft tissue defects proved to be the treatment of choice in perineal procedures. 7. In view of the poor prognosis of conservative methods in chronic acne, and because of the risk of subsequent manifestation of chronic septicemia, early surgical excision is desirable. PMID- 2973186 TI - [Enzymatic determination of plasminogen activators]. PMID- 2973187 TI - Recombinant versus plasma-derived hepatitis B vaccines: issues of safety, immunogenicity and cost-effectiveness. AB - Any attempt to reduce the incidence of hepatitis B on a worldwide scale requires the availability of large quantities of potent, safe and affordable hepatitis B vaccine. However, ongoing doubts or concerns--justified or not--persist about the comparative safety, immunogenicity and cost-effectiveness of commercially available hepatitis B vaccines, whether derived from plasma or produced via recombinant expression systems. This review compares plasma versus recombinant hepatitis B vaccines in terms of these alleged differences and in the light of increasing clinical data acquired following administration of recombinant yeast derived hepatitis B vaccines. PMID- 2973188 TI - Immunogenicity of low doses of recombinant hepatitis B vaccine in young males. AB - A trial of low doses (1.25 and 0.625 micrograms) of recombinant hepatitis B vaccine in 110 males aged 17-19 years is reported. Follow-up was extended to one year and further data are reported for participants from earlier trials of 10, 5 and 2.5 microgram doses for comparison. Seroconversion rates after the booster dose were 94% at 1.25 micrograms and 89% at 0.625 micrograms, compared to 100% at higher doses. Geometric mean titres (GMT) of antibody to hepatitis B surface antigen (anti-HBs) were significantly lower with the 0.625 microgram dose than with 1.25 micrograms at all times, reaching a GMT of 42.6 IU l-1 and 152.9 l-1, respectively, after the booster dose. GMT values at 12 months were 513.9, 510.0, 320.7, 46.5 and 13.3 IU l-1 in the 10, 5, 2.5, 1.25 and 0.625 microgram dose groups, respectively. Increases in GMT after the booster dose were parallelled in all groups. GMT decreased by approximately equal to 65% between 7 and 12 months in the three lowest dose groups, significantly less than the fall of 80% at 5 and 10 micrograms. However, to secure variability among vaccinees, the minimum effective dose for young adults should be greater than 5 micrograms per dose. PMID- 2973190 TI - [Distribution of HLA-DR antigens in the population of Leningrad]. PMID- 2973189 TI - Anti-pre-S2 antibodies in natural hepatitis B virus infection and after immunization. AB - Anti-pre-S2 antibodies were detected by enzyme-linked immuno-absorbant assay using a synthetic peptide analogue of pre-S2 protein, in different groups of hepatitis-B-infected subjects, including patients presenting with cirrhosis and liver cancer, and also in infants immunized with hepatitis B vaccine. Anti-pre-S2 antibodies were not detected in hepatitis B surface antigen (HBsAg) chronic carriers, including patients with cirrhosis or primary liver cancer. Anti-pre-S2 antibodies were not detected in HBsAg-positive sera during the early phase of acute hepatitis. They were only noted upon recovery, when anti-HBs antibodies are detectable at the same time as HBsAg. After recovery, anti-pre-S2 antibodies were noted in 57% of test sera and were still detectable in 16% of anti-HBs-positive sera obtained years after HBV infection. Anti-pre-S2 antibodies were detected in 70% of infants immunized with 2 or 5 micrograms doses of Hevac B Pasteur vaccine, confirming that this vaccine contains pre-S2 antigen. Anti-pre-S2 detection was correlated with the anti-HBs antibody titre. PMID- 2973191 TI - [Prognostic immunologic pretransplantation monitoring in clinical kidney allotransplantation]. PMID- 2973192 TI - Does the repeated orbital sinus puncture alter the occurrence of changes with age in the retina, the lens, or the Harderian gland of laboratory rats? PMID- 2973193 TI - [Experimental infection of pregnant rats with radioactively-labeled Mycoplasma pulmonis]. PMID- 2973195 TI - [Adjuvant gestagen therapy in stage I endometrial cancer]. AB - 221 patients with endometrial cancer stage Ia and Ib have been operated on between 1980 and 1986. 99 patients have been irradiated postoperatively. 122 patients got 300 mg medroxyprogesterone acetate per day over one year provided that the carcinoma was limited to the uterine corpus, the invasion into the myometrium was at a maximum of 2/3 of the uterine wall, there was no invasion of the lymphatic vessels and the tumour was histologic uniform. The 5-year-survival rate of the patients operated on without irradiation but with gestagens was 98.1 per cent calculated with actuarial method in contrast to a matched group from the years 1970 to 1979 with operation and irradiation but without gestagens of 93.2 per cent. These results demonstrate that an individualized therapy in stage I is possible in patients with a low risk of recurrence. PMID- 2973194 TI - Blood pressure and its correlation to selected traits in 2 rat outbred stocks. PMID- 2973197 TI - [Correlation of clinical signs with the outcome of craniocerebral trauma]. AB - Clinical manifestations were correlated with head trauma (HT) outcomes using computer software MEDSTAT-85. Neuro-ophthalmologic signs, motor and vital disorders had maximal prognostic values. Outcome was highly correlated with consciousness disorders phase scaling and patients' state scores (PSS) as well as with coma and consciousness disorders length suggesting that these parameters are of major prognostic importance. A close correlation was found between consciousness states, PSS and HT variants, brain lesion forms, vital and somatic disorders indicating their role as determinants of HT outcome. PMID- 2973196 TI - [A library of genes of plasmid PSS120 of Shigella sonnei]. AB - The gene library of S. sonnei plasmid pSS120 was constructed with the use of plasmid pSL5 as vector. The complete restriction of the vector DNA and the partial restriction of the DNA of plasmid pSS120 were carried out by means of the enzyme EcoRI. The restricted DNA was ligated and packed in vitro into the capsid of phage lambda. The titer of negative colonies obtained after packing was 0.8 X 10(3) clones per 1 microgram of S. sonnei DNA. The total number of detected clones was 250. On the basis of the results, obtained in the analysis of the inserts of the DNA of plasmid pSS120 into the DNA of recombinant clones, the theoretical volume of the library, equal to 92 clones, was calculated. The collection of clones thus obtained will be used for checking the presence of the determinants of invasiveness and phase I antigen, localized in the DNA of plasmid pSS120. PMID- 2973198 TI - [Use of levamisole in the combined therapy of patients with slowly progressing juvenile schizophrenia]. AB - Immunomodulating drug Levamisole was studied as a component in the combined treatment of juvenile slow progredient schizophrenia. Apart from normalizing the immune parameters, the drug improved the patients' clinical conditions. Most prominent was its clinical effect in patients with immune system sensitive to the drug. The patient's condition ameliorated in relation with the features of his initial state. Treating the patients with depressive-asthenic states has yielded the best results. Weaker was the effect on patients with obsessive-phobic disorders. The least curable were senesto-hypochondriac and dysmorphophobic disorders with sensitive delusions of relation. PMID- 2973199 TI - Incidence of thromboembolism in association with congenital disorders in coagulation and fibrinolysis. PMID- 2973200 TI - Drugs recently released in Belgium: goserelin--ofloxacin. PMID- 2973201 TI - Commercially available analogues of GnRH and LH secretion. AB - Six agonistic derivatives of GnRH, four of which have already been evaluated in clinical trials, were compared with GnRH itself in an in vitro test system (incubation of rat pituitary glands). It was investigated 1) how the release of LH was affected when the pituitary glands were incubated in the presence of these analogues or GnRH, and 2) how the release of LH continued after removal of the analogues or GnRH from the medium. It was also investigated how an in vivo pretreatment for 6 days with several doses of 5 of these analogues or GnRH affects 3) the plasma concentration of LH, 4) the pituitary content of LH, and, in vitro, 5) the autonomous and 6) agonist-stimulated secretion of LH. Each of the analogues showed for each of the six investigated parameters a 10- to 100 fold higher potency than GnRH itself. Between the six analogues there were only minor differences. It is discussed how the six investigated parameters may be the expression of one single property of all these analogues, namely a long retention in the pituitary gland with a strong binding to the GnRH receptor. PMID- 2973202 TI - The effect of tenotomy and immobilization on muscle spindles and tendon organs of the rat calf muscles. A histochemical and morphometrical study. AB - The morphological and histochemical alterations in the muscle spindles and Golgi tendon organs of tenotomized or immobilized calf muscles of rats (m. soleus, m. gastrocnemius) were studied in 54 animals. The intact contralateral feet served as controls. There was no change in the number of mechanoreceptors in either the tenotomized or the immobilized muscles. The diameter of the intrafusal fibers was increased somewhat after 1 week, but decreased by 14% to 40% in 3 weeks. The periaxial space of muscle spindle had diminished or disappeared by 2 weeks from the beginning of the procedures. The thickness of the outer capsule of the spindles had markedly increased, especially in the polar region. In the Golgi tendon organs the capsule had also thickened significantly and the internal space diminished. The alterations were similar following tenotomy or immobilization, but more marked after tenotomy than after immobilization. PMID- 2973203 TI - Barriers to utilization of dental health services in a group of disabled Norwegian adults. AB - A group of 496 disabled Norwegian adults were interviewed about their use of the dental health services. The purpose was to study which factors influenced their use of such services. Altogether, 53% of the study participants visited a dentist every year. Among the 335 dentate participants 72% visited the dentist annually, whereas 13% of those without their own teeth reported regular visits. A multiple classification analysis was applied to explain the variance in the use of dental services. Self-reported barriers to dental visits and the number of own teeth left were the strongest and equally important determinants (beta = 43) for regular utilization of dental services when all the participants were included in the analysis. Among the dentate participants self-reported barriers (beta = 55) explained most of the variance in dental visits. PMID- 2973204 TI - [Visual and tactile recognition of words of various lengths and lexical occurrence]. PMID- 2973205 TI - An autopsy case of malignant rhabdoid tumor arising from soft parts in the left inguinal region. AB - An autopsy case of malignant rhabdoid tumor arising from soft parts in the left inguinal region of a 31-year-old Japanese male is reported. The tumor grew rapidly and appeared as a large mass measuring 21.5 X 16.5 cm located in the subcutaneous tissues of the lower abdominal wall. Histologically the tumor was composed of small round cells resembling rhabdomyoblasts, which showed a diffuse or alveolar arrangement. The cytoplasm of the tumor cells was eosinophilic and frequently contained round inclusion bodies, each consisting of an aggregation of globular filaments by electron microscopy. Immunohistochemistry showed the tumor cells to be positive for vimentin and keratin in the cytoplasm, but negative for actin, myoglobin and myosin. At autopsy, extensive metastatic or invasive tumors were observed mainly in the retroperitoneum and pelvic organs. The patient showed hypercalcemia which disappeared for only short periods just after surgery. PMID- 2973206 TI - The endogenous concentration and subcellular distribution of androgens in normal human premenopausal endometrium, myometrium and vagina. AB - The endogenous concentrations of testosterone (Testo), androstenedione (Adion), 5 androstene-3 beta, 17 beta-diol (Adiol), dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEAS) have been measured in endometrium, myometrium and vagina from 23 premenopausal women at different stages of the cycle. The tissue to plasma gradient was positive for all androgens except DHEAS; a significant correlation existed between plasma and tissue levels of Adiol and DHEAS. In all tissues significant correlations were calculated between Adiol and DHEA, Adion and Testo, and Adiol and DHEAS. Except for Testo, androgen concentrations were different in the tissues with a significant correlation between the tissues, each tissue seeming to be able to create its own specific intratissue androgen pattern. During the cycle Testo was highest in myometrium and vagina and Adion in myometrium and endometrium during the secretory phase. Except for DHEA, androgens were preferentially localized in the cytosol fraction of the cells. From this localization and the known inhibitory effects of androgens on the activity of estrogen metabolizing enzymes, it is postulated that androgens play a role in enzymatic rather than receptor-mediated processes in uterine cells. PMID- 2973207 TI - Trisomy 21. Molecular and cytogenetic studies of nondisjunction. PMID- 2973208 TI - Molecular aspects of receptors and binding factors for IgE. PMID- 2973209 TI - Further advantages of the "vest over pants" abdominoplasty. AB - The standard abdominal lipectomy is a risky procedure with respect to where the first incision (i.e., the transverse suprapubic incision) is located. Failure to place the incision successfully may distort the whole procedure, resulting in an overly tight abdominal wall, hypertrophy, and upward displacement of the scar along with the pubic hair, and also excessive tension causing skin necrosis of the abdominal flap. Advantages of our reversed technique are emphasized. PMID- 2973210 TI - Laser treatment of rhinophyma. AB - The carbon dioxide laser, when used in the treatment of rhinophyma, allows accurate removal of the diseased tissue to the desired level without risk of exposure of any underlying osteocartilaginous structures. This ensures rapid reepithelialization that minimizes the untoward scarring that may occur from delayed healing using other sculpturing techniques. Precise tissue vaporization prohibits violation of surrounding normal tissues. PMID- 2973211 TI - Prolonged disability from job-related injury. AB - Workers who do not recover as expected from job-related injuries are frequently disabled not by organic complications of their injuries but by a problem that the patient may not acknowledge and the physician may not recognize and deal with. The family physician should be skilled in detecting nonorganic causes of disability and in intervening appropriately to maintain a therapeutic relationship between the patient and the health care providers. PMID- 2973212 TI - Influence of coronary collaterals on left ventricular function in patients undergoing coronary angioplasty. AB - The purpose of this study was to determine the role of coronary collaterals on global left ventricular function during transient coronary occlusion. Left ventricular systolic function was evaluated noninvasively in 37 patients undergoing percutaneous transluminal coronary angioplasty (PTCA) by means of peak aortic blood acceleration measured with a continuous-wave Doppler velocity meter placed suprasternally. Doppler measurements were made before, during, and immediately after balloon inflation. Nineteen patients underwent PTCA of the left anterior descending coronary artery, 15 of the right coronary artery, and three of the circumflex coronary artery. All patients had a subtotal coronary occlusion. Among the 37 patients 23 had no angiographic evidence of collaterals supplying the vessel undergoing PTCA and 14 had collaterals. In patients without collaterals peak acceleration was 16.0 +/- 3.0 m/sec/sec before balloon occlusion and decreased to 11.8 /+- 3.6 m/sec/sec within 30 to 40 seconds of coronary occlusion (p less than 0.001). After 30 to 40 seconds of reperfusion (balloon deflated), peak acceleration returned to near- preocclusion levels (17.3 +/- 7.5 m/sec/sec). Among patients with collaterals peak acceleration remained unchanged during balloon occlusion and reperfusion relative to preocclusion levels (NS). These data indicate that collaterals can preserve left ventricular function during transient coronary occlusion in humans. PMID- 2973213 TI - Assessment of coronary angioplasty by an automated digital angiographic method. AB - Digital subtraction coronary angiograms (DSA) of 63 patients who had undergone coronary angioplasty (PTCA) for a total of 73 lesions were analyzed with an automated border-detecting computer program capable of simultaneous geometric and densitometric cross-sectional area estimation. The computer measurements were compared with visual interpretation of the 35 mm cineangiograms. The results indicated that visual reports of cineangiograms tend to overestimate the pre-PTCA diameter percent stenosis and to underestimate the post-PTCA residual stenosis in comparison with the computer (p less than 0.001 in bot cases). There was good agreement between geometric and densitometric area percent stenoses calculated by the program on the pre-PTCA digital angiograms (r = 0.82, p less than 0.001, mean of their differences = -0.2 with standard deviation = 6.1). Following PTCA, however, important discrepancies between the two methods existed (r = 0.71, p less than 0.001, mean of their differences = 1.0 with standard deviation = 18.6). Following PTCA (but not pre-PTCA), densitometric evaluation demonstrated a significantly greater mean coefficient of variation between different views (69%) than did the geometric evaluation on the same views (24%). We conclude (1) that visual interpretation of cine coronary angiograms compares poorly with quantitative methods for both the selection of PTCA candidates and the assessment of the results; (2) that the geometric and densitometric characteristics do not agree in describing the degree of post-PTCA residual stenosis; and (3) that after angioplasty, important discrepancies between densitometric evaluation in different views are observed. PMID- 2973214 TI - Increased circulating atrial natriuretic factor (ANF) release during induced ventricular tachycardia. AB - During sustained ventricular tachycardia (VT), there is usually a rise in mean atrial pressure. Atrial natriuretic factor (ANF) is a peptide released by the atria in response to increased atrial pressure or distension. ANF causes arterial vasodilation and natriuresis in man, and may contribute to the polyuria noted during tachycardia. We hypothesized that induction of sustained VT would cause elevation of right atrial pressure and lead to increased release of ANF. We measured mean arterial pressure, right atrial pressure, VT cycle length, atrial cycle length, plasma norepinephrine, and central ANF in 11 patients during 20 episodes of VT. The mean cycle length of induced tachycardia was 335 +/- 15 msec, with a mean atrial cycle length of 547 +/- 43 msec. Baseline plasma ANF was 70 +/ 11 femtomoles per milliliter and increased to 133 +/- 22 fmol/ml (p less than 0.001) after 4 minutes of VT. There was a decrease in mean arterial pressure from 87 +/- 3 to 66 +/- 3 mm Hg, and a rise in mean right atrial pressure from 4.0 +/- 0.8 to 11 +/- 1 mm Hg (p less than 0.001). The change in plasma ANF was quite variable among individual patients and was correlated with changes in mean right atrial pressure but poorly correlated with changes in mean arterial pressure, plasma norepinephrine, and atrial cycle length. Our data demonstrate that significant increased in ANF levels occur during sustained VT and are associated with increased right atrial pressure. PMID- 2973215 TI - Clinical significance and prognostic importance of left ventricular hypertrophy in non-Q-wave acute myocardial infarction. AB - Left ventricular (LV) hypertrophy is known to be an independent risk factor for cardiac death, but its significance in non-Q-wave acute myocardial infarction (AMI) has not been assessed previously. In a randomized diltiazem-placebo controlled therapeutic trial of non-Q-wave AMI confirmed by creatine kinase-MB (CK-MB), 126 of 544 patients (23%) exhibited LV hypertrophy using standard voltage criteria. Compared to patients without LV hypertrophy, patients with LV hypertrophy were significantly older (65 vs 60 years, p less than 0.0001) and had smaller peak adjusted CK levels (490 +/- 376 vs 666 +/- 726 IU/liter, p less than 0.001) than patients without LV hypertrophy. Patients with and without LV hypertrophy did not differ significantly in acute mortality during hospitalization, progression to Q waves, reinfarction by CK-MB criteria or angina associated with transient electrocardiographic changes. Compared with patients without LV hypertrophy, those patients with non-Q-wave AMI and LV hypertrophy had a 2-fold higher incidence of reinfarction (24 vs 12%, p less than 0.005) and death (19 vs 9%, p = 0.044) during the first year of follow-up. Multivariate regression analysis revealed that the relative risk of death and reinfarction during the initial year after AMI was increased by a factor of 1.7 and 2.1 among patients with LV hypertrophy, respectively. It was therefore concluded that, although patients with LV hypertrophy and non-Q-wave AMI have smaller enzymatic infarcts and the same short-term prognosis as do patients without LV hypertrophy, their reinfarction and mortality rates are significantly increased during the first year of follow-up.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973216 TI - Location of an acute myocardial infarct in patients with a healed myocardial infarct: analysis of 129 patients studied at necropsy. AB - To determine the relation of a single healed myocardial infarct to a fatal acute myocardial infarct, 129 patients with 1 grossly visible healed and 1 grossly visible acute infarct were studied at necropsy. It was determined whether the acute infarct was opposite to or adjacent to the healed infarct or if 1 infarct was so large that it was both opposite to and adjacent to the other infarct. In 74 (57%) of the 129 patients, the 2 infarcts were opposite one another, in 40 (31%) they were adjacent and in 15 (12%) they were both opposite and adjacent. The age, sex, mean size of the healed infarct and heart weight were similar among the 3 groups. Acute myocardial infarcts were larger in the group that had both opposite and adjacent infarcts than either of the other 2 groups (p less than 0.001). Information regarding whether the infarcts were clinically recognized or not was available in 108 patients: both infarcts were recognized in 41 (38%), neither infarct was recognized in 15 (14%) and 1 infarct was recognized and the other was not in 52 (48%). The number of the 4 major epicardial coronary arteries narrowed at some point greater than 75% in cross-sectional area by atherosclerotic plaque was similar in patients with recognized and in those with unrecognized infarcts. Similar numbers of narrowed major epicardial coronary arteries also were found in each of the 3 infarct groups (opposite, adjacent or both). PMID- 2973217 TI - Transient alterations of the QRS complex and ST segment during percutaneous transluminal balloon angioplasty of the left anterior descending coronary artery. AB - Using continuous 3-lead electrocardiographic (ECG) recordings in 19 patients undergoing elective percutaneous transluminal coronary artery angioplasty (PTCA) of the left anterior descending (LAD) artery, this study described the dynamic changes of the ST segment and the R- and S-wave amplitudes that occur during transient myocardial ischemia. The waveforms from lead V2 were quantified at 10 second intervals during the length of the balloon inflation that produced the greatest extent of ST-segment deviation. The simultaneous changes that occurred in leads aVF and V5 were also observed, but not quantified. Measurements of R- and S-wave amplitudes were performed during maximal ischemia from both the PR- and the J-ST-segment baselines to determine which of these most nearly maintained its control position during ischemia. The results indicate that the R-wave amplitude is best determined from the PR-segment baseline (p = 0.0007), while the S wave is best determined from the J-ST-segment baseline (p = 0.03). However, only a portion of the QRS changes observed during PTCA could be accounted for by the baseline shift. There were additional QRS changes during ischemia in 11 of the patients (58%) suggestive of conduction disturbances in 3 endocardial sites: left septal, right septal and left anterosuperior. It is hypothesized that these changes may represent ischemia-induced delay in conduction ("periischemic block") previously thought to occur only with myocardial infarction. PMID- 2973218 TI - Hemodynamic, humoral and volume findings in systemic hypertension with isolated ventricular septal hypertrophy. AB - To analyze the hemodynamic, endocrine and volume characteristics of isolated septal hypertrophy (ISH) in established systemic hypertension, 22 patients with ISH were compared to 23 patients with symmetric hypertrophy and to 28 without left ventricular (LV) hypertrophy. Mean arterial pressure and 24-hour ambulatory pressure readings did not differ between the 2 groups. At the same level of arterial pressure, patients with ISH had a high cardiac index (p less than 0.02) and a faster heart rate (p less than 0.05); consequently, total peripheral resistance was decreased (p less than 0.05). Although there was no change in intravascular volume, central blood volume was expanded (p less than 0.02), and the ratio of central to peripheral blood volume was increased (p less than 0.02), thereby indicating peripheral venoconstriction. Patients with isolated ISH had greater responses of diastolic pressure and heart rate (p less than 0.05) to isometric stress than the other 2 groups. A hyperdynamic circulatory state is a hemodynamic hallmark of ISH in early essential hypertension that might be produced by increased sympathetic activity. PMID- 2973220 TI - Ventricular excitation during percutaneous transluminal angioplasty of the left anterior descending coronary artery. PMID- 2973219 TI - Effects of isradipine, a new calcium antagonist, versus hydrochlorothiazide on serum lipids and apolipoproteins in patients with systemic hypertension. AB - The effect of isradipine versus hydrochlorothiazide on the lipid profile of 44 hypertensive patients was investigated in a double-blind, randomized, 2-center trial. Lipid profiles included total cholesterol, serum triglycerides, high density lipoprotein (HDL) cholesterol, HDL subclasses, (HDL2 and HDL3), low density lipoprotein cholesterol, very low density lipoprotein cholesterol, apolipoprotein A-1 and apolipoprotein B. Isradipine had no effect on the lipid profile in short- (4 and 10 week) or long-term (52 week) studies. Hydrochlorothiazide increased serum triglycerides in 11 of 13 patients by a mean of 8% for the group (p less than 0.05) in long-term (52 week) studies, and total cholesterol by a mean of 9 and 16%, respectively (p less than 0.01) in 2 of 13 patients, with no difference in other lipid or lipoprotein parameters in short- or long-term studies. PMID- 2973221 TI - Atrial natriuretic peptide in acute myocardial infarction. PMID- 2973222 TI - Usefulness of echocardiographic left ventricular hypertrophy, ventricular tachycardia and complex ventricular arrhythmias in predicting ventricular fibrillation or sudden cardiac death in elderly patients. PMID- 2973223 TI - Left ventricular hypertrophy in cyclosporine-induced systemic hypertension after cardiac transplantation. PMID- 2973224 TI - Pregnancy following percutaneous transluminal angioplasty. PMID- 2973225 TI - Distribution of microspheres in plantaris muscles of resting and exercising rats as a function of fiber type. AB - The purpose of this study was to determine histologically the distribution of microspheres (MSs) (14 micron), and hence the relative distribution of blood flow, in rat plantaris muscle relative to the fiber types (fast-twitch-oxidative glycolytic [FOG], fast-twitch-glycolytic [FG], and slow-twitch-oxidative [SO]). Three conditions were investigated: 1) preexercise standing; 2) treadmill locomotion at 15 m/min (fast walking); and 3) treadmill locomotion at 60 m/min (moderate galloping). The MS suspension (containing 1 x 10(6) MSs) was infused into the ascending aorta via a catheter in the carotid artery under each of the 3 conditions so that MSs were distributed to the tissues in proportion to their respective blood flows. Sections (20 micron) of the plantaris muscle were cut and assayed for reduced nicotinamide adenine dinucleotide tetrazolium reductase (NADH TR) and myofibrillar adenosine triphosphatase (ATPase) activities so the fibers could be typed as SO, FOG, or FG. MSs were located in the NADH-TR sections, and the fibers next to the MSs were classified and counted. The observed numbers of fibers of each type in each condition that were adjacent to MSs were compared to the predicted number of adjacent fibers based on the assumption the MSs were randomly distributed in the tissue. This analysis demonstrated that MSs (and blood flows) were preferentially distributed to SO fibers during preexercise, to SO and FOG fibers during slow locomotion, and to FOG fibers during fast locomotion. The data support the contention that blood flow is distributed in muscles of conscious animals as functions of fiber type and exercise intensity. PMID- 2973226 TI - ECOG phase II trials of MGBG, chlorozotocin, COM multidrug therapy in advanced measurable colorectal cancer. AB - The Eastern Cooperative Oncology Group (ECOG) entered 326 patients with advanced measurable colorectal cancer into four phase II drug or drug combination trials. Previously treated and chemotherapy-naive patients were eligible. Chlorozotocin was administered to 83 patients (51 previously treated), methyl-glyoxal-bis guanylhydrozone (MGBG) to 90 patients (58 previously treated), and two regimens of the three-drug combination of cyclophosphamide, vincristine, and methotrexate (COM) to 153 patients (120 previously treated). The multidrug regimen had been developed specifically for previously treated patients. In this trial, chemotherapy-naive patients were no more likely to respond than were members of the previously-treated group. Even among previously untreated patients, response rates did not exceed 10% in any of these phase II programs. They are not recommended for further trials in patients with colorectal cancers. PMID- 2973227 TI - The effect of temperature change of composites on the bonding strength of orthodontic brackets. AB - In an orthodontic office, the bonding system is subjected to certain temperature variation that ranges between 5 degrees C (refrigeration) and 25 degrees C (room temperature). The purpose of this study was to determine whether such temperature variation has any effects on tensile and shear bond strengths and the type of bracket failure. Two groups of 20 teeth each were compared; in one group the temperature of the composite during bonding was 5 degrees C and in the other group the temperature was 25 degrees C. Student's t test was used to determine whether significant differences were present between the two groups. The results of the present findings indicate that the variation in the temperature of the composite, within the limits tested, does not seem to adversely affect the bonding strength of the bracket to the enamel. In addition there was no difference between the two groups in the type of interface bond failure among the enamel, composite, and bracket. PMID- 2973228 TI - Impaired echinocytic transformation of ankyrin- and spectrin-deficient erythrocytes in mice. AB - The membrane skeleton of the red blood cell plays an important role in the determination of cell deformability and cell shape. Under various in vitro conditions, red blood cells undergo an echinocytic or stomatocytic shape transformation. The mechanism of this fundamental process is not well understood. We have studied the red cell shape transformation in normoblastic anemia mice (nb/nb) and spherocytic anemia mice (sph/sph), which are deficient in ankyrin and spectrin, respectively. We found that both ankyrin-deficient cells (nb/nb) and spectrin-deficient cells (sph/sph) have a reduced capacity to undergo echinocytic transformation with various echinocytogenic treatments, that is, incubation with sodium salicylate (40 and 120 mM), calcium loading (50 microM A23187 + 2.2 mM Ca2+), or metabolic depletion (24 hr at 37 degrees C). These results suggest that the functional integrity of the membrane skeleton is essential for the maintenance and transformation of the red cell shape. PMID- 2973229 TI - Improved predictive testing for Huntington disease by using three linked DNA markers. AB - Eighty-five persons at risk for Huntington disease (HD) have enrolled in a predictive-testing pilot program. Informativeness of the test has been determined for 41 of these candidates by using linked DNA probes. Nine (21.9%) of these persons have been excluded from the test as a result of the unavailability of DNA from crucial family relatives. Homozygosity for all of the three DNA markers (D4S10, D4S62, and D4S95) was not found in any affected parent. Only one (2%) of the 41 test candidates has had an uninformative result. Results have been given to 20 persons, of whom 12 (60%) received a decreased risk and eight (40%) received an increased risk of having inherited the mutant gene for HD. The combined use of three DNA markers significantly increases the informativeness of family structures such that some change in the estimation of genetic risk is now possible for approximately 75% of all persons who request predictive testing. PMID- 2973230 TI - Huntington disease in Georgia: age at onset. AB - Age at onset of motor symptoms was collected on 611 persons affected with Huntington disease (HD) among 3,201 persons "at risk" in 108 kindreds. Life-table estimates correcting for truncated intervals of observation (censoring) produced a median age at onset 5 years older than the observed mean. Risk estimates of HD onset for persons at risk, as calculated by life-table methods, were significantly higher for older ages than were estimates based on the observed distribution of onsets. Age-specific incidence was found to be highest at age 35 64 years, a considerably older age interval than suggested by previous estimates. The offspring of affected males had significantly younger onset than did offspring of affected females, and a trend suggesting and excess of paternal descent among juvenile-onset cases was present. Life-table analysis is contrasted with analyses of (a) the observed distribution of age at onset and (b) remote cohorts age 63 or older at the time of data collection. The implications for risk prediction, genetic counseling, and genetic analysis of HD are discussed. PMID- 2973231 TI - Health and socioeconomic consequences of occupational respiratory allergies: a pilot study using workers' compensation data. AB - Growing evidence suggests that individuals with occupational allergies have long term health and employment problems. Workers' compensation claims for allergic respiratory disease allowed in Ontario 1975-1981 were reviewed for demographic, risk factor, and exposure data. Diagnoses listed in these 244 claims were reassessed for consistency with criteria for occupational respiratory allergies. Twenty-eight percent of the 154 claimants who met the criteria were granted a permanent disability award from the workers' compensation board (WCB). In response to a questionnaire mailed 2-8 years after claim allowance, 77% of those traced reported improvement, but 59% still required medication and 85% still suffered symptoms. Seventy-six percent of claimants left their employer. Seventy five percent of those who left did so due to their allergic condition; 95% suffered long-term income loss. The unemployment rate for this cohort was 36%, well above that for the general population. Older workers with longer duration of symptoms and longer duration of exposure prior to the claim had the worst prognoses. The data were consistent with previous studies in suggesting that occupational respiratory allergies may result in serious health and socioeconomic consequences. Exploring the feasibility, acceptability, and potential usefulness of WCB data, it is suggested that this source merits further consideration both for descriptive as well as prognostic studies. PMID- 2973232 TI - Antibodies and immune profiles of individuals occupationally exposed to formaldehyde: six case reports. AB - Six patients with multiple subjective health complaints, which have been correlated with chronic exposure to formaldehyde during the course of their education and occupations, were tested for the existence of antibodies (IgE, IgM, and IgG) to formaldehyde (F) conjugated to human serum albumin (F-HSA). In addition, the percentage and absolute numbers of peripheral lymphocyte subpopulations as determined by surface markers were investigated. Antibody titers to F-HSA were present as follows: IgE (2 patients), IgM (3 of 4 tested patients), and IgG (5 patients). Analysis of lymphocyte subpopulations showed T helper/suppressor (H/S) ratios ranging from 0.8 to 3.3. All 6 patients had elevated Tal cells (antigen memory cells), whereas interleuken 2 receptor positive cells were within expected values. Following formaldehyde exposure, 5 of the patients complained of an initial flulike illness from which they have not completely recovered. The sixth individual had a history of recurrent respiratory infections and surgical removal of hyperplastic ethmoid sinus tissue. The common occurrence of anti-F-HSA antibodies, flulike illness, and Tal cells are interpreted as suggestive of a chronic antigenic stimulation of the immune system in these 6 patients. Further immunological work-up of additional subjects and immune parameters with similar history of formaldehyde exposure and subjective health complaints is warranted. PMID- 2973233 TI - Digital and palmar dermatoglyphics in dementia of the Alzheimer type. AB - Digital and palmar dermatoglyphics were examined in 29 men and 27 women with dementia of the Alzheimer type (DAT) and 112 age-, sex-, and racial group-matched controls. Female patients had significantly (p less than 0.05) more accessory triradii and complete Sydney creases than controls; no dermatoglyphic differences were detected in the males. Separating the patients by age of onset prior to or after age 65 years did not help differentiate patients from controls by dermatoglyphic profile. This study failed to confirm either the previously reported dermatoglyphic differences between DAT patients and controls or the reported similarity of the dermatoglyphic pattern of DAT to that of Down syndrome patients. PMID- 2973234 TI - Holoprosencephaly and trisomy 21 in a child born to a nondiabetic mother. AB - This is the second reported case of a child with holoprosencephaly and trisomy 21. The first case was born to a diabetic woman; in our case, there was no evidence of diabetes in the mother. Most of the distinctive facial features of Down syndrome were obscured by the presence of cyclopia and a supraorbital proboscis in this infant. The relevance of chromosome analysis in cases with holoprosencephaly is discussed. PMID- 2973235 TI - Chance vs. causality in the association of Down syndrome and holoprosencephaly. PMID- 2973236 TI - Fetal hypoxia elevates plasma atrial natriuretic factor concentration. AB - Acute hypoxia in the fetus is associated with a reduction in fetal blood volume. We hypothesized that atrial natriuretic factor in the fetal circulation may be one of the factors that mediate this blood volume decrease. Thus the present study was designed to determine the effects of hypoxia on circulating concentrations of atrial natriuretic factor in fetal sheep and correlate these changes with fetal blood volume. Hypoxia was induced in chronically catheterized sheep fetuses by infusing nitrogen containing CO2 into the trachea of the ewe for 30 minutes. Fetal arterial PO2 decreased by 10.2 +/- 1.3 (SE) mm Hg. Plasma atrial natriuretic factor concentration rose concurrently with the fall in PO2 such that atrial natriuretic factor increased to 565 +/- 196 pg/ml from a basal level of 127 +/- 13 pg/ml (p less than 0.001). Fetal blood volume was reduced by 7.2% +/- 2.1% and was significantly related to changes in atrial natriuretic factor levels (p less than 0.0001). At the termination of hypoxia, PO2 returned to normal levels before plasma concentrations of atrial natriuretic factor fell to baseline values. Therefore fetal hypoxia appears to be a potent stimulus for elevating plasma concentration of atrial natriuretic factor in the fetus, and this rise in atrial natriuretic factor in the circulation may be partially responsible for the reduction in fetal blood volume observed during hypoxia. PMID- 2973237 TI - Prenatal diagnosis in metropolitan Atlanta and the impact on autosomal trisomies. PMID- 2973238 TI - Infantile glaucoma in Down's syndrome (trisomy 21) PMID- 2973239 TI - Occupational therapy knowledge: from practice to theory. 1988 Eleanor Clarke Slagle lecture. AB - This paper discusses the place of occupational therapy technology in practice and the growth of scientific technology. It is suggested that practice is an important source for the development of technological theory. A historical review of the treatment of hand dysfunction in cerebral palsy is presented as an illustration of the development of a specific occupational therapy technology. PMID- 2973240 TI - Distribution of red cell phosphoglucomutase-1 subtypes in several Mongoloid populations of East Asia. AB - The distribution of red cell phosphoglucomutase (PGM) subtypes was determined by starch-gel electrophoresis and isoelectric focusing in a group of 2,484 unrelated individuals from ten Mongoloid populations of East Asia. The sample comprised 998 Chinese from various localities--Singapore, 325; Malaysia, 270; Taiwan, 276; Hong Kong, 67; Fouzhou, 60--as well as 342 Koreans; 252 Filipinos; 529 Thais; 336 Malays, and 27 Indonesians. Altogether 15 phenotypes controlled by four common and five rare alleles at the PGM1 locus were observed in these populations. The frequency of the most frequent allele (PGM1+) varied from 0.56 to 0.74, with the highest frequency observed in the Singapore Chinese and the lowest in the Malays. Within the Chinese from different localities a significant degree of heterogeneity was observed at the PGM1 locus. The rare allele (PGM17)6 was observed only among the Chinese, Thais, and Malays, while the PGM1 was lacking in the Filipinos. A new allele with ahigh pI (6.5) was observed in a low frequency in all the populations but the Malays. PMID- 2973241 TI - Coordination of electromechanical and metabolic properties of cat soleus motor units. AB - Motor units were studied in the soleus muscle of normal adult cats and adult cats that had undergone complete spinal cord transection approximately 4 mo earlier. Intracellular recording and stimulation techniques were used to study selected electrical properties of the motoneuron and isometric contractile properties of the muscle unit. Motor unit fibers were depleted of their glycogen through repetitive stimulation of the motoneuron and identified by a quantitative histochemical determination of glycogen. A sample of muscle fibers from the glycogen-depleted unit and from fibers not depleted of glycogen were analyzed for cross-sectional area, succinate dehydrogenase (SDH), alpha-glycerolphosphate dehydrogenase (GPD), and alkaline myofibrillar adenosine triphosphatase. It was observed that the fiber-to-fiber variability in cross-sectional area and SDH and GPD activity within units of normal and transected cats was significantly larger than that measured in repeated samples from a single fiber. Additionally, for each of these properties, the range found among fibers within a unit was similar to that found among nondepleted fibers of the same myosin type. The influence of spinal cord transection on some muscle fibers seemed to result in a metabolic shift from the generalized category of slow-oxidative toward fast-oxidative glycolytic. This shift in metabolic properties appeared to be coupled with a similar shift in the physiological properties of the muscle unit and motoneuron from slow to fast. PMID- 2973242 TI - ANF inhibits glucose-stimulated insulin secretion in mouse and rat. AB - Atrial natriuretic factor (ANF) is synthesized in atrial cells and was recently demonstrated to also occur within islet glucagon cells. Therefore, we investigated the possible effects of synthetic rat ANF-(1-28) on basal and stimulated insulin and glucagon secretion in the mouse and on glucose-induced insulin secretion in the rat. We found that ANF did not affect basal levels of insulin, glucagon, or glucose when injected intravenously at dose levels between 0.25 and 4.0 nmol/kg in mice. However, when injected together with glucose (2.8 mmol/kg), ANF (4.0 nmol/kg) inhibited the increase in plasma insulin levels by 40%, from 114 +/- 8 microU/ml in controls to 81 +/- 8 microU/ml (P less than 0.01). Likewise, the increase in plasma insulin levels during an intravenous infusion of glucose in rats (10 mg/min) was significantly reduced by ANF (100 pmol.kg-1.min-1; P less than 0.001). In contrast, the increase in plasma levels of insulin and glucagon after the intravenous injection of the cholinergic agonist carbachol in mice (0.16 mumol/kg) was not significantly affected by ANF. We conclude that ANF inhibits glucose-stimulated insulin secretion in the mouse and the rat. The peptide may therefore be a modulator of insulin secretion. PMID- 2973243 TI - Differential localization of 5-HT1 receptors on myenteric and submucosal neurons. AB - Intracellular recordings were made from guinea pig enteric neurons, and the effects of 5-hydroxytryptamine (5-HT) and the 5-HT1 selective agonists 5 carboxyamidotryptamine (5-CT) and 8-hydroxy-2-(n-dipropylamino)tetralin (DPAT) were studied on membrane potential and synaptic potentials. Most myenteric AH neurons were hyperpolarized when these agonists were applied by superfusion; this hyperpolarization was due to an increase in potassium conductance. Membrane hyperpolarizations to 5-HT, 5-CT, or DPAT were never observed in submucous neurons. Fast nicotinic excitatory postsynaptic potentials (EPSPs) and slow EPSPs recorded from S neurons in the myenteric plexus were suppressed by 5-HT, 5-CT, and DPAT; slow EPSPs in myenteric AH neurons were also inhibited by these agonists. Fast and slow EPSPs recorded from submucous S neurons were not affected by 5-CT or DPAT. However, slow EPSPs recorded from submucous AH neurons were readily blocked by 5-CT and DPAT. The results indicate that 5-HT1 receptors are located on the cell bodies of myenteric but not submucosal neurons. The nerve terminals that release the mediator or mediators of fast and slow synaptic potentials in myenteric neurons also have 5-HT1 receptors and presumably arise from other myenteric neurons; the nerve terminals responsible for the slow EPSP to AH neurons seem to be the only elements of the submucous plexus that express 5 HT1 receptors. PMID- 2973245 TI - Effects of a physiological dose of ANP on renal function in dogs. AB - The current study was designed to evaluate the effects of a physiological dose (1.0 ng.kg-1.min-1 into the renal artery) of atrial natriuretic peptide (ANP, 8 33) on renal function in anesthetized mongrel dogs. Left renal blood flow (RBF) was measured with a flow probe and the glomerular filtration rate (GFR) using both clearance and renal extraction methodologies. After a 60- to 90-min stabilization period and collection of two 5-min control clearances, ANP was infused for 20 min, with four 5-min clearances performed. The increase in ANP concentration in renal arterial plasma (calculated from the ANP infusion rate and the renal plasma flow) was 212 +/- 9 (SE) pg/ml. During each experimental period there was a saliuresis and diuresis, an increase in the reabsorption of solute free water, and an increase in the creatinine clearance; mean arterial blood pressure and the GFR, calculated from extaction data, were unaffected by ANP.RBF was 3.28 +/- 0.21 during control and 3.03 +/- 0.22 ml.min-1.g-1 at 20 min during ANP infusion (P less than 0.01). These data demonstrate that a physiological dose of ANP is natriuretic and diuretic, causes a modest decrease in RBF without affecting the GFR, and causes a condition in which clearance methods can overestimate the GFR. PMID- 2973244 TI - Arginine vasopressin promotes growth of rat glomerular mesangial cells in culture. AB - Arginine vasopressin (AVP) binds specifically to vascular smooth muscle-like mesangial cells (MCs) and affects contraction. We tested whether this peptide also modulates growth behavior of rat MCs in early subculture (passage 2-5). Subconfluent, serum-starved MCs were exposed to AVP (10(-10)-10(-6) M) in the presence or absence of insulin (5 micrograms/ml). To assess DNA replication, MC uptake of [3H]thymidine (24-h pulse) was determined on days 1, 2, and 3. AVP alone averaged a 1.97-fold increase in DNA synthesis at 24 h, whereas the mean stimulatory effects of AVP at 48 and 72 h were 7.21- and 5.42-fold, respectively. MCs exposed simultaneously to AVP and insulin showed potentiation of the mitogenic response to AVP alone. The V1-receptor antagonist [1-(beta-mercapto beta,beta-cyclopentamethylene proprionic acid), 2-(O-methyl-Tyr)-Arg]vasopressin (PMP) inhibited only AVP-induced promotion of MC growth (maximal inhibition of 78.3%). The phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA) acutely stimulated MC proliferation but did not add to the AVP effect. Preincubation of MCs with 600 nM of TPA for 48 h significantly inhibited AVP-induced mitogenesis ( 87.2%). By use of fura-2, intracellular calcium (Cai) was assessed by spectrofluorometry. The addition of AVP (10(-12)-10(-6) M) led to a rapid, transient, dose-dependent increase in Cai of 154-383%, respectively. The AVP induced increase in Cai was greatly inhibited by 3,4,5-trimethoxybenzoic acid 8 (diethylamino)octyl ester hydrochloride (TMB-8) (10(-8)-10(-6) M), an inhibitor of Cai release (-23.9 to -72.1%), and it was blunted by the atrial natriuretic peptide AP-28 (-38.3%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973246 TI - ANF secretion and renal responses to volume expansion with equilibrated blood. AB - To evaluate the role of atrial natriuretic factor (ANF) in the renal response to acute blood volume expansion without hemodilution, a reservoir syringe filled with donor rat blood was connected to the femoral artery and vein of anesthetized Sprague-Dawley rats to allow rapid equilibration of the reservoir with the intravascular blood. Volume expansion with blood from the reservoir in two steps (of 1 and 1.5% body wt, separated by 1 h, n = 5 rats) produced a mean peak increase in plasma immunoreactive ANF from 99 +/- 21 to 1,310 +/- 230 pg/ml (P less than 0.001); plasma ANF levels throughout these experiments correlated significantly with simultaneously measured urine flow (r = 0.74, P less than 0.005) and sodium excretion (r = 0.65, P less than 0.005). Another group (n = 7) underwent the same two-step procedure; after the second volume expansion, high dose atriopeptin III infusion (0.4 microgram.kg-1.min-1 did not further increase fractional excretion of sodium (3.17 +/- 0.27 to 2.50 + 0.39%, P = NS). In another group (n = 9 rats), the same dose of atriopeptin III was started before any blood volume expansion. After the resulting hypotension was corrected by restoration of blood volume, an additional 1.5% body weight blood volume expansion did not further augment sodium excretion. We conclude that the diuresis and natriuresis, which occur in response to volume expansion without hemodilution, rise and fall in parallel with immunoreactive ANF in the plasma, and that ANF and acute blood volume expansion act on the kidney through a similar, saturable mechanism. PMID- 2973247 TI - Dopamine1 receptors in rat kidneys identified with 125I-Sch 23982. AB - Dopamine1 receptors were studied in rat kidney using the selective dopamine1 antagonist 125I-labeled Sch 23982. The specific binding of 125I-Sch 23982 (defined by 5 microM Sch 23390) to renal cortical homogenates incubated at room temperature was rapid, saturable with time and ligand concentration, and reversible. Analysis of Rosenthal plots revealed a single class of receptors with an apparent dissociation constant of 12.2 +/- 1.9 nM and maximum receptor density of 1.03 +/- 0.15 pmol/mg protein (n = 6). However, competition experiments with the dopamine1 antagonist Sch 23390 revealed a low- and high-affinity binding site with inhibition constants of 1 x 10(-6) and 1 x 10(-8) M, respectively. The competition experiments were also indicative of dopamine1 receptors with stereoselectivity noted for dopamine1 but not for dopamine2 antagonists. The inhibition constants for dopamine1 antagonists and agonists were two orders of magnitude greater in renal cortical than striatal homogenates. Different buffers affected striatal but not renal cortical binding. Autoradiographic studies revealed 125I-Sch 23982 binding in renal cortical but not medullary tissue. These studies confirm the presence of dopamine1 receptors in the cortex of the rat kidney. PMID- 2973248 TI - Intramural distribution of intestinal blood flow during sympathetic stimulation. AB - Recent studies indicate that the mucosal circulation of the small intestine possesses more potent local circulatory control mechanisms than the muscularis. Several lines of evidence support this assertion: the absence of reactive hyperemia in the muscularis, the confinement of glucose-induced hyperemia to the mucosal circulation, and the more effective autoregulation of villus blood flow in comparison with total blood flow. Therefore, we postulated that the mucosal circulation would exhibit a more pronounced ability to escape the vasoconstrictor influence of sympathetic nerve stimulation. To test this hypothesis, we used laser-Doppler velocimetry (LDV) to study the effects of perivascular nerve stimulation on blood flow in the muscularis and mucosa of isolated dog intestine. In two series of experiments, we measured total blood flow to a gut loop with an electromagnetic flow probe on the supply artery, while LDV measurements were made either on the mucosa or on the muscularis. Sympathetic stimulation (10 Hz) transiently reduced total blood flow to approximately 10% of control in both studies. Muscularis and mucosal blood flow both reached minimal values. Subsequently, total blood flow and the two regional perfusions exhibited "autoregulatory escape," but the propensity for blood flow to escape from sympathetic vasoconstriction was significantly greater in the mucosa than in the muscularis. These data provide more evidence that, compared with the muscularis, the intestinal mucosa possesses more potent local control mechanisms that oppose neurogenic vasoconstriction. PMID- 2973249 TI - Regional hemodynamic effects of atrial natriuretic peptide or captopril in Brattleboro rats. AB - The regional hemodynamic effects of rat atrial natriuretic peptide (ANP) (1 and 10 nmol) were measured in the same conscious Brattleboro rats in water-replete and in water-deprived states (14 h). In the latter condition, the effects of ANP were compared with those of captopril (3.2 mumol). In rats in the water-replete state, the low dose of ANP had no significant effects on mean arterial pressure (MAP) or renal, superior mesenteric, or hindquarter vascular resistances, although heart rate (HR) showed a transient increase. The high dose of ANP caused a fall in MAP and an increase in HR. Renal and hindquarter vascular beds showed dilatation followed by constriction; there was a mesenteric vasoconstriction only. In rats in the water-deprived state, the low dose of ANP caused a marked fall in MAP but only a transient increase in HR; there was marked hindquarters vasodilation. The high dose of ANP caused dramatic hypotension and bradyarrhythmia; an initial hindquarter vasodilation gave way to vasoconstriction; renal and mesenteric vasoconstrictions also occurred. The initial effects of captopril on MAP were similar to those of the high dose of ANP, but the regional hemodynamic effects of captopril indicated a potent inhibition of both primary and secondary vasoconstrictor mechanisms. PMID- 2973250 TI - Organ-specific control of glycolysis in anoxic turtles. AB - Control of glycolysis during anoxia was investigated in five organs (heart, brain, liver, and red and white skeletal muscles) of the freshwater turtle, Pseudemys scripta, after 1 or 5 h of submergence in N2-bubbled water. Lactate was produced as the metabolic end product, with distinct organ differences in the amount (net lactate accumulation was 2.4-fold higher in brain than white muscle) and rate (lactate production in liver dropped 16-fold after the 1st h) of lactate accumulation. ATP and total adenylate contents of all organs were reduced (by 15 32%) after 1 h of submergence, but energy charge was maintained; after 5 h, adenylate contents had fully recovered. Changes in the levels of hexose and triose phosphate intermediates of glycolysis indicated an activation of glycolysis within the 1st h of anoxia exposure in brain, heart, and skeletal muscles. By 5 h, however, these were reversed, and a glycolytic rate depression was indicated, consistent with the overall metabolic rate depression accompanying long-term anaerobiosis in the turtle. Crossover analysis indicated glycolytic control at the pyruvate kinase reaction in all organs during both glycolytic activation and metabolic depression; regulatory control at the phosphofructokinase locus was primarily important only during glycolytic activation in heart and red muscle. The same analysis indicated a very rapid glycolytic inhibition in liver occurring within the 1st h of anoxia exposure; this allows glycogenolysis to be directed toward glucose export yielding the fermentative fuel used by other organs during anoxia. PMID- 2973251 TI - Fetal and adult urine flow and ANF responses to vascular volume expansion. AB - Isotonic saline (2% of body wt) warmed to body temperature was infused intravascularly over 10 min into chronically catheterized, near-term fetal sheep and into nonpregnant adult sheep. Although both groups of animals underwent the same initial increase in urine flow rate relative to body weight, the diuresis was significantly more prolonged in the adult compared with the fetus. The adult demonstrated a sustained increase in plasma atrial natriuretic factor (ANF) concentration with a suppression of plasma renin activity (PRA) and plasma arginine vasopressin (AVP). In the fetus, only a transient rise in plasma ANF with no change in PRA or AVP occurred. The increases in blood volume and arterial pressure were significantly greater in the adult compared with the fetus, but the venous pressure changes were the same. Thus it appears that fetal and adult kidneys have the same initial urine flow response to rapid vascular volume loading presumably due to a pressure diuresis and elevations in ANF. The more prolonged diuresis in the adult appears attributable to more extensive and prolonged changes in plasma hormone concentrations. The latter, in turn, may be due to the greater intravascular retention of the infused fluid in the adult than in the fetus. PMID- 2973252 TI - Mechanisms of active suppression of the immune response to spermatozoa. AB - The production of autoantibodies to spermatozoa in males and isoantibodies in females is inhibited both by the physical isolation of spermatozoa from the systemic immune system and by active immunosuppression mechanisms. Lymphoid cells present in the epithelial lining of the rete testis, epididymis, and vas deferens, as well as the human ejaculate, are predominantly T suppressor/cytotoxic cells. Mononuclear cells derived from semen inhibit the in vitro activation of peripheral blood lymphocytes. Soluble specific T suppressor/cytotoxic cell activators in semen or on the sperm surface may be responsible for the predominance of this T cell subset in the male reproductive tract. The activation of T suppressor/cytotoxic cells following coitus may also limit the immune response to spermatozoa in females. Spermatozoa can also initiate immunosuppression, either by selectively inducing T suppressor cells or through the generation of activated complement components that block antibody production. Antisperm antibodies in sera from females may be associated with either a deficiency in the ability of their T suppressor/cytotoxic cells to be induced by factors in semen or by the occurrence in their husbands' ejaculates of microorganisms, antibodies, or other factors that induce T helper lymphocytes. Activated T cells produce interferon gamma, which induces Ia antigen expression on macrophages and allows the female's T helper cells to recognize processed sperm antigens. Recognition of the role of cell-mediated immune functions in the male and female genital tract identifies possible new target sites for the development of contraceptive agents. PMID- 2973253 TI - Low back pain among retired wrestlers and heavyweight lifters. AB - The lifetime incidence and prevalence of low back pain among 32 retired wrestlers (ages 39 to 62 years) and 13 retired heavyweight lifters (ages 40 to 61 years) were evaluated and compared to the corresponding results in a cross-sectional study of 716 men (ages 40 to 47 years). The radiologic findings and the findings upon physical examination in the athletes were compared to the findings in another study of normal, active, similarly aged men who were sampled at random. The lifetime incidence and prevalence of low back pain was higher among the wrestlers (59%) compared with both the lifters (23%) and the control group (31%). The tolerance for backache seemed to be higher among the athletes than the controls. A higher frequency of old fractures was found among the wrestlers. The athletes with fractures had a higher frequency of low back pain. A significant decrease in disk height was found among the lifters. PMID- 2973254 TI - New perspectives on anesthetic agents. AB - The practices of anesthesiology are changing to meet the needs of an older and sicker patient population, to adapt to advances in medical technology and operating techniques, and to provide better postoperative conditions for patients and those caring for them. There is a large spectrum of anesthetics and a variety of anesthetic techniques that allow anesthetic onset and recovery to be rapid, vital functions to be maintained within normal limits, and operating conditions to be adapted to the needs of both patients and surgeons. Nevertheless, all anesthetic drugs and techniques have inherent risks as well as benefits, and despite expertise and caution in their use, a satisfactory outcome for the patient cannot be guaranteed. PMID- 2973255 TI - Prognostic implication of cellular DNA content in acute lymphoblastic leukemia. AB - To assess the prognostic value of the cellular DNA content measured by flow cytometry in acute lymphoblastic leukemia (ALL), we studied the pretreatment distribution of the DNA content in marrow blasts from 74 children with this disease. They were divided into standard- and high-risk groups according to their white blood cell counts and age at the time of diagnosis and were followed for up to 44 months. Abnormal DNA stemlines were detected in 26 children, and all of them had a hyperdiploid DNA content (DNA index of greater than 1.0). The duration of remission was significantly longer in those with hyperdiploid DNA stemlines than in those with diploid DNA stemlines in both risk groups. In the standard risk group, the DNA content and common ALL antigen were significant prognostic factors, especially the former. In the high-risk group, the DNA content also showed an independent significant correlation with the clinical outcome. PMID- 2973256 TI - Immunological changes associated with a successful outcome of pollen immunotherapy. AB - Changes in immunological measurements thought to be important in the mechanism of immunotherapy for hay fever were related to objective measurements of treatment outcome. Antigen-specific IgE and IgG, and T suppressor cell status, using monoclonal antibodies and a specific functional assay, were measured before and after immunotherapy. Treatment outcome was assessed using nasal and conjunctival challenge tests. Seventeen subjects received immunotherapy for 6 months. Nine showed a decrease in nasal and/or conjunctival sensitivity, compared with 2 of 13 control subjects (P less than 0.05). Subjects responding to immunotherapy had a significantly higher post treatment IgE level and a higher pre- and post treatment IgG level than non-responders and controls. Immunotherapy had no effect on suppressor cell status. PMID- 2973257 TI - A continuous high flow intermittent mandatory ventilation system incorporating a Downs venturi device and a modified Nuffield Series 200 ventilator. AB - Systems for respiratory support are becoming increasingly expensive and complex. Many systems suffer inadequacies when used for spontaneous ventilation. Some modes on newer ventilators are rarely used because of the complex controls and settings. There is no truly universal ventilator that satisfies every intensivist's wishes. CPAP/IMV is becoming accepted as the standard management of many patients with acute respiratory failure and there would be few intensive care units where CPAP/IMV is not used for part of a patient's respiratory support. We describe a cost-effective system that may be used for respiratory support in the spontaneously breathing mode. This system combines a high flow venturi, an efficient humidifier and an inexpensive reliable ventilator that can be used for adult and paediatric patients. The system, primarily for use in patients breathing spontaneously, functions well in patients requiring full ventilation. PMID- 2973258 TI - Oxygen rotameters--left- or righthand drive? PMID- 2973259 TI - A plasma membrane-enriched preparation from giant barnacle muscle fibers. AB - We describe a procedure for obtaining a highly enriched plasma membrane (sarcolemmal) preparation from muscle fibers of the giant barnacle (Balanus nubilus). The sarcolemmal-enriched portion migrated as a light fraction (F1) at the 10-24% sucrose interface. This fraction displayed saturable ouabain binding (Kd = 0.119 microM) that was enriched 10 times compared to that in the original homogenate. F1 was also prepared using muscle fibers previously labeled with 1,2 ditritio-1,2(2,2'-disulfo-4,4'-diisothiocyano)diphenylet hane, disodium salt [( 3H]-H2DIDS). F1 was enriched 25-fold in [3H]H2DIDS binding sites with respect to the homogenate. Ca2+-ATPase and succinic dehydrogenase-activities were low in F1, as was oxalate-supported Ca2+ uptake. Compared to membranes of sarcoplasmic reticulum origin, F1 was enriched in sarcolemma membranes by about 45-fold while it was enriched approximately 30-fold over mitochondrial membranes. Thus, F1 provides an extremely pure source of external muscle membranes. PMID- 2973260 TI - An automated Western blot analysis using the phastsystem. AB - We have developed a rapid, automated method for preparing Western blots of very small amounts of proteins, utilizing a commercially available electrophoresis system (Phastsystem, Pharmacia). This system has been adapted to transfer to nitrocellulose experimental samples that were initially separated in the same system by gradient-sodium dodecyl sulfate gel electrophoresis. The developing unit of the Phastsystem has permitted automation of all the necessary steps including incubation with antibodies, saturation of nonspecific binding sites, and washing. The total elapsed time for protein separation, transfer, and staining is about 6 to 7 h. PMID- 2973261 TI - Method for identification of intracellular free flavin species in the photosensitive fungus Neurospora crassa. AB - Establishing the relative intracellular proportions of flavins in Neurospora crassa (and in other organisms) in vivo may be hampered by degradation of flavins after homogenization of the cells. The system described here allows separation and identification of intracellular free and bound flavins under conditions restrictive for the FAD-degrading enzyme(s). A "protective buffer" containing 0.1 M citrate adjusted to pH 4.0 with K2HPO4, 5 mM ATP, and 0.5 mM EDTA prevents FAD from rapid enzymatic cleavage in crude cell lysates of the Neurospora crassa mutant "slime." PMID- 2973262 TI - High affinity antibody from hen's eggs directed against the human insulin receptor and the human IGF-I receptor. AB - Large quantities of high affinity antibodies directed against the human insulin receptor and the human insulin-like growth factor-I (IGF-I) receptor were obtained from hen's eggs. Hens were immunized with human placental membranes and human liver membranes by intramuscular injections. Specific antibodies to the receptors were demonstrated in serum and egg yolks at 5 weeks and these antibodies presisted for at least 6 months. Antibodies from egg yolks were purified by the polyethylene glycol precipitation technique of Polson et al. The eggs provided the equivalent of about 450 ml of immunized serum per month per bird. The purified antibodies were approximately equally reactive with receptors for insulin or IGF-I. Antibodies immunoprecipitated affinity-labeled receptors, inhibited binding of each ligand, and were capable of stimulating 2-deoxyglucose uptake in rat adipocytes and thymidine incorporation in cultured fibroblasts. The presence of antibodies directed against the IGF-I receptor in those hens immunized with human liver membranes was unexpected, since liver membranes possess little or no IGF-I binding activity. We conclude that antibodies against human antigens may be relatively easily obtained by immunization of hens and purification of those antibodies from eggs. PMID- 2973264 TI - Electron microscopic cytochemical localization of Ca-ATPase in the rod outer segments of the toad Bufo marinus. AB - Transmission electron microscopy and ultracytochemistry were employed in an attempt to localize the enzyme calcium adenosine triphosphatase (Ca-ATPase) in the rod outer segments (ROS) of the toad retina. Utilizing a one-step incubation procedure, Ca-ATPase was identified as an electron-dense precipitate in the intradiskal spaces of the rod disks (vesicles) of the ROS. Analytical microscopy identified the reaction product as lead phosphate. The formation of the reaction product was dependent on the presence of ATP (the substrate) and calcium ions. However, calcium ions could be substituted for by magnesium ions. In addition, the reaction was vanadate sensitive. The latter is known to inhibit Ca-ATPase activity. Such data appear to indicate the presence of a Ca-Mg-ATPase in association with the rod disks. Since cyclic guanosine monophosphate (cyclic GMP), rather than calcium ions, is currently believed to be the primary intracellular messenger associated with phototransduction, the presence of an ROS Ca-ATPase may indicate other functions for this cation in the physiology and biochemistry of the visual process. Ca-ATPase might play a role in directional calcium fluxes between intracellular compartments. PMID- 2973263 TI - Cellular immunofluorescence: quantification of low abundance proteins. AB - Aminofluorescein can be extracted with alkaline carbonate buffer (0.5% Na2CO3 in 0.1 M NaOH) from fixed cells stained in indirect immunofluorescence by fluorescein-conjugated antibody. Fluorescence is then quantified by spectrofluorometry. A standard curve obtained by dilution of known fluorochrome allows for subsequent spectrofluorometric analysis of the extracted aminofluorescein. A saturating quantity of primary antibody should be used to determine the level of staining associated with a cellular antigen. This simple method makes it possible to quantify samples used for immunofluorescence microscopy. It can be adapted for determining the DNA content in the same samples, allowing the quantity of antigen to be equated either to DNA content or to cells plated. PMID- 2973265 TI - Endocytosis of parotid salivary proteins by striated duct cells in streptozotocin diabetic rats. AB - Salivary gland striated duct cells play an important role in the modification of primary saliva by secretion and reabsorption of electrolytes, and secretion of glycoproteins. Recent observations have shown that in the rat parotid gland these cells are able to internalize exogenous proteins, e.g., horseradish peroxidase and ferritin, from the ductal lumen. In rats made diabetic by injection of streptozotocin, dense vacuoles and crystalloids are present in the apical cytoplasm of parotid striated duct cells. In this study we utilized electron microscopic immunocytochemistry to determine if these vacuoles and crystalloids contain acinar secretory proteins. At various times after induction of diabetes by streptozotocin (65 mg/kg), the parotid glands were fixed in a glutaraldehyde formaldehyde mixture, postfixed in OsO4, and embedded in epoxy resin. Thin sections were immunolabeled with antibodies to protein B1 (Ball et al., 1988) and alpha-amylase (Baum et al., 1982) using a modification of the Protein A-gold technique (Bendayan and Duhr, 1986). With antibody to B1, label was localized in the secretory granules of acinar and intercalated duct cells of both normal and diabetic rats. In striated duct cells of diabetic rats, label was present over the electron-dense vacuoles but not over the crystalloids. Since crystalloids appear to form within the vacuoles, their lack of reactivity may indicate degradation of the internalized protein. The same distribution of label was found with antibody to amylase except for the intercalated duct granules, which were unlabeled in both control and diabetic animals. These results demonstrate that striated duct cells take up salivary proteins from the lumen and that the endocytosis of some secretory proteins from the saliva may be a significant function of these cells in certain pathological conditions. PMID- 2973266 TI - Respiratory effects of epidural and subcutaneous morphine, meperidine, fentanyl and sufentanil in the rat. AB - This study compared the respiratory effects of subcutaneous and epidural morphine, meperidine, fentanyl, and sufentanil in rats breathing air or 8% CO2 in air. A whole body plethysmographic technique was used to measure minute volumes of breathing. The ED50s of subcutaneously injected morphine, meperidine, fentanyl, and sufentanil in depressing the minute volume response to 8% CO2 in air were 2300 micrograms/kg, 8800 micrograms/kg, 20 micrograms/kg, and 2.3 micrograms/kg, respectively. These doses were nearly the same as the subcutaneous ED50s of these compounds in producing analgesia, found in an earlier study. Roughly equianalgesic doses of the four opiates after epidural injection, however, failed to cause any detectable respiratory effect. Fourfold greater doses increased significantly the incidence of low minute volumes with fentanyl and sufentanil, but soon after epidural injection, i.e., at the time that analgesia was produced. None of the epidurally injected opiates had a significant delayed effect on respiration. However, one of the seven rats treated epidurally with the higher dose of morphine developed depression of the minute volume response to 8% CO2 in air as late as 7 hours after the injection. We conclude that epidural injection, in contrast to subcutaneous injection, of analgesic doses of morphine, meperidine, fentanyl, and sufentanil produces no significant respiratory effects. PMID- 2973267 TI - The dose-effect relationship of metocurine: EMG versus MMG. PMID- 2973268 TI - Electromagnetic field focusing probe (EFFP)--a new angioplasty tool. AB - An electromagnetic field focusing probe (EFFP) consists of a radiofrequency generator, solenoidal coil, and a hand-held or catheter probe. The probe is operated in the near field (distance within one wave length of an electromagnetic field source) of a coil, which induces eddy current in a biological tissue. The induced eddy current is converged maximally at the tip of the probe upon contact of the tip with the tissue. The probe produces very high temperatures depending on the wattage selected. In this study, the EFFP was used to evaporate atheromatous plaques in human cadaver abdominal aorta specimens, which were then studied histologically. Gas produced by this technique was analyzed and the volume found to be related to power delivered, but in such small amounts as to be of no embolic significance. While temperature varied with wattage and time of application, it was maximal at the probe tip and easily controlled, resulting in clean obliteration of plaque. PMID- 2973269 TI - A placebo-controlled comparison between the bronchodilatory effects of ipratropium bromide inhaled as a dry powder and by metered dose inhaler in chronic obstructive pulmonary disease. AB - A placebo-controlled study was performed to compare the effect of the inhalation of ipratropium bromide as a powder (capsule = 40 micrograms) and by pressurized aerosol (two puffs of 20 micrograms; ie, 40 micrograms). Fifteen patients (nine males and six females) with chronic obstructive pulmonary disease were studied in a double-blind crossover comparison of the two different modes of administration. The VC, FEV1 and viscous work of breathing time-response curves were almost identical, indicating bronchodilation. We conclude that in patients with chronic obstructive pulmonary disease, the powder inhalation was not more effective than the pressurized aerosol. It could, however, be offered as an alternative to patients with poor hand-lung coordination. The patients tolerated the two modes of administration without difficulties: no local irritation or coughing was observed. PMID- 2973270 TI - Experience with the use of tPA in the treatment of acute myocardial infarction. AB - Early experience with the use of tissue plasminogen activator (tPA) in acute myocardial infarction is reviewed, including comparisons with other thrombolytic agents, a summary of hemorrhagic complications associated with its use, and the rationale for adjunctive therapeutic strategies. The use of tPA has been associated with improvement in left ventricular function, a lower mortality, and a decrease in congestive heart failure signs and symptoms. A protocol for evaluation of patients with possible myocardial infarction for thrombolytic therapy is presented. Consideration must be given to other possible diagnoses, and the ECG must be evaluated carefully to ensure that appropriate criteria are met. Risk factors for hemorrhagic complications include recent trauma, surgery, gastrointestinal and genitourinary bleeding, stroke, and focal neurologic findings. Greater benefit of therapy is expected in patients with larger infarcts who have more marked ST segment changes or evidence of hemodynamic compromise, especially when they are treated early after the onset of symptoms (within the first several hours). Adjunctive measures that can be considered in the emergency department include prophylactic lidocaine, IV nitroglycerin, beta blockade, aspirin, volume replacement and monitoring for dysrhythmias, bleeding, and recurrent ischemia. A comprehensive understanding of these rapidly evolving concepts will assist the emergency physician in the evaluation and management of patients with acute myocardial infarction. PMID- 2973271 TI - Virulence factors and markers in Escherichia coli from calves with bacteremia. AB - Relative pathogenicity of 151 Escherichia coli isolates from 36 calves with bacteremia after necropsy was studied by measurement of the LD50 after mice were inoculated IP with E coli isolates. Study of virulence factors and markers revealed that the pathogenicity of E coli was associated with the production of hydroxamate siderophores and with resistance to serum bactericidal effects. Production of colicins, including colicin V, and of surface antigen 31A was correlated with virulence. The close association between phenotypic expression of virulence factors and markers was consistent with a hypothesis of a localization of genes coding for virulence factors and markers on the same plasmid. PMID- 2973272 TI - Traumatic abdominal hernia: a case report and review of the literature. AB - Traumatic abdominal hernia remains a rare clinical entity despite an overall increase in blunt abdominal trauma. What appears to be the most extensive traumatic abdominal hernia so far described is presented. Traumatic abdominal herniae fall into three general categories: small lower quadrant abdominal defects and inguinal hernias, typically the result of blunt trauma with bicycle handlebars, are the most common; larger abdominal wall defects sustained in motor vehicle accidents are the next most common hernias; intra-abdominal herniations through rents in the retroperitoneum are rarely seen. The diagnosis may often be established with physical examination alone. Conventional radiology, computerized tomography, and ultrasound have also proven useful. Because of the high incidence of other associated intra-abdominal injuries, early exploration and repair through a midline incision is advocated. Adequate debridement and solid repair of fascial planes with non-absorbable sutures are required to prevent recurrence. PMID- 2973273 TI - [Esophageal achalasia: apropos of a case of Down's syndrome]. PMID- 2973274 TI - Elevated plasma atrial natriuretic factor and vasopressin in high-altitude pulmonary edema. AB - A diagnosis of acute high-altitude pulmonary edema was made in five male skiers (age, 35.0 +/- 1.8 years) by history and physical examination and was confirmed by a characteristic chest radiogram showing alveolar infiltrates associated with a normal cardiac silhouette. Five healthy age- and sex-matched subjects with similar physical activity at the same altitude served as controls. Plasma sodium was 135.0 +/- 1.5 mmol/L in the acutely ill patients compared with 144.0 +/- 3.3 mmol/L in the controls (P less than 0.025). Mean plasma atrial natriuretic factor immunoreactivity averaged 17.6 +/- 5.6 pmol/L in patients with high-altitude pulmonary edema compared with 6.8 +/- 0.7 pmol/L in the controls at the same altitude (P less than 0.05). Elevated atrial natriuretic factor levels normalized to 7.5 +/- 1.9 pmol/L (P less than 0.05) during recovery in Denver (altitude, 1600 meters) 24 hours later. Plasma arginine vasopressin levels were 1.8 +/- 0.37 pmol/L in patients with high-altitude pulmonary edema at diagnosis compared with 0.92 +/- 0.28 pmol/L in controls (P = 0.07). The inappropriately elevated arginine vasopressin levels decreased to 1.29 +/- 0.37 pmol/L during recovery (P less than 0.025), but the lowered plasma sodium concentration had not normalized by discharge within 24-hours of transfer to Denver and averaged 135.8 +/- 1.2 mmol/L. The pathophysiologic implications of these findings are discussed. PMID- 2973275 TI - Nonspecific interstitial pneumonitis without evidence of Pneumocystis carinii in asymptomatic patients infected with human immunodeficiency virus (HIV). AB - STUDY OBJECTIVE: To assess how often Pneumocystis carinii organisms, P. carinii pneumonia, or other pulmonary pathologic processes were present in persons infected with human immunodeficiency virus (HIV) without pulmonary symptoms or previous history of P. carinii, and with a normal chest roentgenogram. DESIGN: Serial, prospective assessment of eligible HIV-seropositive patients over 21 months. PATIENTS: Twenty-four HIV-seropositive patients with either a nonpulmonary manifestation of the acquired immunodeficiency syndrome (AIDS) (n = 12) or an absolute CD4 lymphocyte count of 0.200 X 10(9) cells/L or less (n = 12), no pulmonary symptoms, a normal chest roentgenogram, no history of P. carinii pneumonia, and no history of treatment with antipneumocystis prophylaxis. INTERVENTIONS: Pulmonary assessment with arterial blood gases, pulmonary function tests, gallium-67 citrate scans, and bronchoscopy with bronchoalveolar lavage and transbronchial biopsies. MEASUREMENTS AND MAIN RESULTS: Mean alveolar-arterial gradient was 11.1 mm Hg +/- 8.5 and mean diffusion capacity was 73.0% +/- 20.0% of predicted. None of the 24 patients showed P. carinii or other pathogens on stains of bronchoalveolar lavage fluid. No patient had histologic evidence of P. carinii pneumonia. Transbronchial biopsy specimens showed chronic, nonspecific interstitial pneumonitis (11 of 23) and no pathologic abnormality (12 of 23). Six patients have developed P. carinii pneumonia during 2 to 18 months of follow-up. CONCLUSIONS: HIV-infected patients without pulmonary symptoms did not have detectable Pneumocystis organisms in bronchoalveolar lavage fluid or transbronchial biopsy specimens; but 11 of 23 had evidence of chronic, nonspecific interstitial pneumonitis. Pneumocystis organisms in a pulmonary specimen from a symptomatic patient probably indicate the cause of the pulmonary dysfunction even if only a few are detected. PMID- 2973276 TI - The physiopathological role of quinolinic acid in the mammalian central nervous system. PMID- 2973277 TI - Influence of antiestrogen drugs on the sex hormone and sex hormone-binding globulin levels in breast cancer patients. PMID- 2973278 TI - Surgical management of persistent congenital eversion of the upper eyelids. AB - Most cases of congenital eversion of the upper eyelids resolve spontaneously within two weeks of birth. We present a case of persistent upper-eyelid eversion that did not even after two months of conservative management. A previously undescribed mechanism, consisting of a vertical elongation of the eyelid in its posterior aspect creating a relative skin deficiency, was probably responsible for this eversion. Skin grafts provided rapid resolution with a good cosmetic result. PMID- 2973279 TI - A stoma jig for surgical craft workshops. AB - A new jig simulating the abdominal cavity and wall is described. The techniques of defunctioning colostomy and closure of colostomy, end colostomy and ileostomy can be performed. If required the techniques of mass or layered closure of an abdominal incision can also be practised. PMID- 2973280 TI - Treatment of abdominal nerve entrapment syndrome using a nerve stimulator. PMID- 2973281 TI - Diagnosis of significant abdominal trauma after road traffic accidents. PMID- 2973282 TI - [Direct cutaneous ureterostomy with myoplasty using a pedicled flap of the rectus abdominis]. AB - Many methods have been reported to prevent the development of stomal stenosis following direct cutaneous ureterostomy, thus avoiding catheterizations with their attendant complications, and improving patient tolerance. Nevertheless, although lessened, the risk of stenosis persists in narrow ureters. In order to improve local trophicity and help maintain a wide, flexible intraparietal segment, the authors have developed a technique of cutaneous ureterostomy using a muscle flap from the rectus abdominis muscle and a V-shaped skin flap. This procedure was used on 20 ureters, including 12 dilated ureters and 8 narrow ureters. Follow-up was 3 months to 2 years. In 18 cases, no catheterization was needed, and no stenoses or complications were observed. In one case, stomal stenosis developed on a dilated, irradiated ureter from a non-functioning kidney. An indwelling catheter was used in one narrow ureter with a stenosis proximal to the abdominal wall. These preliminary results suggest that this technique is beneficial. Further studies with a larger number of patients and longer follow ups are needed. PMID- 2973283 TI - In vitro susceptibilities of mycoplasmas and ureaplasmas to new macrolides and aryl-fluoroquinolones. AB - In vitro activities of the new macrolides clarithromycin, previously designated A 56268 (TE-031), and A-63075 and of the aryl-fluoroquinolones difloxacin (A-56619) and temafloxacin (A-62254) against 14 strains of Mycoplasma pneumoniae, 20 strains of Mycoplasma hominis, and 28 strains of Ureaplasma urealyticum were compared with that of erythromycin. All three macrolides inhibited growth of M. pneumoniae at less than 0.125 micrograms/ml. No macrolide was active against M. hominis. For five strains of U. urealyticum, MICs were greater than 256 micrograms/ml for all 3 macrolides. Excluding these, no other strain of U. urealyticum had an initial MIC of clarithromycin of greater than 1 microgram/ml, while five had initial MICs of erythromycin which were greater than 4 micrograms/ml. A-63075 was the least active of the three macrolides against ureaplasmas. Temafloxacin and difloxacin had similar activities against all three species, initially inhibiting 90% of M. pneumoniae strains at 2 and 8 micrograms/ml, 90% of M. hominis strains at 2 and 4 micrograms/ml, and 90% of U. urealyticum strains at 4 and 8 micrograms/ml, respectively. Additional pharmacokinetic and clinical trials with the new macrolides and quinolones with mycoplasmal or ureaplasmal infections are indicated. PMID- 2973284 TI - Pharmacokinetics of teicoplanin in pediatric patients. AB - The pharmacokinetics of teicoplanin have been studied in 13 pediatric male patients from 2 to 12 years of age. Patients were given a single 3-mg/kg intravenous dose of teicoplanin for prophylaxis. Blood and urine samples were collected for 8 days after administration, and teicoplanin levels were determined by microbiological assay. Pharmacokinetic parameters were estimated from a three compartment open pharmacokinetic model and from a noncompartmental analysis. Levels in plasma 1 h after the administration averaged 14.8 mg/liter. The half lives of the two distribution phases were 1.3 and 9.7 h. The half-life of the terminal phase averaged 57.9 h, with similar estimates obtained from the noncompartmental analysis and from data from urine. The volume of distribution of the central compartment was 0.15 liter/kg, whereas the volume of distribution at steady state and during the elimination phase were 0.80 and 1.25 liters/kg. The total teicoplanin clearance averaged 14.8 ml/h per kg, with renal clearance accounting for about 60% of the total. The average cumulative recovery of teicoplanin in urine over 8 days was 59% of the dose, similar to the value obtained in adult volunteers. There was no significant linear correlation between elimination half-life and age. Preliminary data after repeated administration support the reliability of the model used and the validity of the mean estimated parameters. There were no local or systemic adverse reactions to teicoplanin. PMID- 2973285 TI - Effects of aminoglycosides and spectinomycin on the synthesis and release of lipopolysaccharide by Escherichia coli. AB - The effects of aminoglycosides and spectinomycin on lipopolysaccharide (LPS) synthesis and release by Escherichia coli were studied. LPS synthesis was previously reported to be regulated by the stringent control mechanism. In agreement with this, the control of LPS synthesis in amino acid-deprived relA+ cells was relaxed by spectinomycin, a proven stringent control antagonist, but not by kanamycin, an agent which is ineffective as a stringent control antagonist. The other stringent control antagonists tested (gentamicin, tobramycin, and, to a lesser extent, amikacin) unexpectedly failed to relax the control of LPS synthesis, and this was subsequently shown to be due to their inhibitory action on LPS synthesis. The release of LPS by nongrowing (amino acid deprived and antibiotic-treated) bacteria was stimulated only under conditions in which the control of LPS synthesis was relaxed. PMID- 2973286 TI - Acitretin (RO10-1670) and oral contraceptives: interaction study. PMID- 2973287 TI - A clinical survey of aortobifemoral bypass using two inherently different graft types. AB - The performance of knitted Dacron and polytetrafluoroethylene (PTFE) bifurcated grafts are compared in this study of 312 patients at a single institution. Patients of the two graft groups were statistically well-matched in risk factors and degree of distal obstructive disease. Operating time needed to implant either graft was approximately equal. For patients with abdominal aortic aneurysms, mean volume of blood transfused was 2.2 units for Dacron grafts and 0.2 units for PTFE grafts; for patients with aortoiliac occlusive disease, the comparable figures were 1.1 units and 0.1 units, respectively. Four-year cumulative patency for Dacron (90%) and PTFE (97%) grafts were not significantly different (p greater than 0.01). Complications affected 13% of the patients of the Dacron group and 4% of the PTFE group. All six graft infections and all seven graft double-limb thromboses occurred in Dacron grafts. Anastomotic aneurysms, amputations, and late graft revisions occurred with greater frequency in patients with Dacron grafts. PMID- 2973288 TI - Porcine-valved Dacron conduits in Fontan procedures. AB - From a series of 52 Fontan procedures between 1976 and 1984, the cases of the 27 consecutive patients who received a porcine-valved conduit were reviewed. There were 5 hospital deaths among these 27 patients. Follow-up ranges from 11 years 9 months to 3 years 9 months. At follow-up, no conduit-related complications could be demonstrated. There were no signs of valvular stenosis, exuberant peel formation, or calcification of the conduit in any of the patients. To date, there has been no need to replace any of the porcine-valved conduits. Cumulative survival (including hospital deaths) is 71% at 10 years. In conclusion, we believe that the porcine-valved conduits have functioned very satisfactorily over time. PMID- 2973289 TI - [Immunomodulating action of a preparation of yeast dsRNA on cellular immunity in mice]. AB - The effect of a dsRNA preparation (an interferon inductor) on DTH induced by SRBC was studied. It was shown that at optimal antigenic load the dsRNA preparation inhibited DTH whereas at suboptimal and supraoptimal loads the preparation stimulated it. The findings indicated that the dsRNA preparation had an immunoregulatory effect. The immunoregulatory properties of the preparation must be associated with its action on the lymphocyte suppressor cells and macrophages. PMID- 2973290 TI - Specific binding and steroidogenetic effects of atrial natriuretic factor (ANF) in Leydig cells of rats. AB - Atrial natriuretic factors (ANF) may influence testicular steroidogenesis. This study was conducted to evaluate the presence of specific ANF-binding on isolated adult rat Leydig cells and the effects of ANF on testosterone production. Indirect immunofluorescence technique demonstrates that adult rat Leydig cells possess specific ANF-binding and that rAP-II strongly stimulates the testosterone production. rAP-II exerts its maximal stimulatory effect on the testosterone secretion at low doses (10(-11) M), corresponding at the physiological plasmatic concentration in the adult normal rat. High doses (10(-9)-10(-7) M) of rAP-II show a decline in the stimulatory effect on testosterone secretion. Our data suggest that rAP-II influences the testicular steroidogenesis by a receptorial mechanism; the biphasic effect of rAP-II on the testosterone production may be related to an acute receptorial desensitization phenomena. PMID- 2973291 TI - [Report on coronary angioplasty by the joint working group of the International Society and Federation of Cardiology and the World Health Organization]. PMID- 2973292 TI - [Atrial natriuretic factor]. AB - The atrial natriuretic factor represents a group of atrial peptides structurally and functionally related. These peptides produce the following effects: natriuresis, diuresis, vasodilation and inhibition of aldosterone release. Its discovery in 1981 opened a new horizon in the studies on body sodium and water homeostasis, as well as on blood pressure regulation. In a few years there have been spectacular advances in the knowledge of this new hormone. However, there are still many doubts and controversies concerning its exact physiological role. Its effects on renal function, vascular contractility and its relationship with other physiological systems are not completely understood. The versatility of functions in which atrial natriuretic factor participates has pointed new trends in the research of important clinical entities such as arterial hypertension and cardiac failure, and could be a starting point for the development of novel therapeutic agents. In this work, the experimental findings dealing with the participation of atrial natriuretic factor in several biological events are reviewed. PMID- 2973293 TI - [Atrial natriuretic factor, catecholamines and the renin-angiotensin system in cardiac insufficiency. Relation to hemodynamic parameters]. AB - Plasma concentrations of atrial natriuretic factor (ANF), catecholamines (adrenaline, noradrenaline, dopamine) and aldosterone, and plasma renin activity (PRA) were measured in a group of 20 patients with moderate to medium heart failure (NYHA class II 7 patients, class III 13 patients), 24 hours after treatment was discontinued. Compared with a control group, plasma concentrations of ANF (p less than 0.01), noradrenaline (p less than 0.05), aldosterone (p less than 0.01) and PRA (p less than 0.01) were significantly increased. There was a significant difference between class II patients and class III patients in plasma ANF (p less than 0.01) and noradrenaline (p less than 0.02) concentrations, but not in PRA and aldosterone levels. A significant correlation was observed between plasma ANF concentration and left ventricular end-diastolic pressure (r = 0.68, p less than 0.001), pulmonary arterial pressure (r = 0.59, p less than 0.01), pulmonary capillary pressure (r = 0.51, p less than 0.02), cardiac index (r = 0.46, p less than 0.05) and left ventricular end-diastolic volume (r = 0.50, p less than 0.05). However, ANF concentration was not correlated with mean right atrial pressure. Plasma adrenaline concentration correlated with systemic arterial resistance (r = 0.80, p less than 0.001), pulmonary arterial pressure (r = 0.57, p less than 0.02), mean pulmonary capillary pressure (r = 0.62, p less than 0.001), cardiac index (r = 0.53, p less than 0.05) and left ventricular end diastolic pressure (r = 0.58, p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973294 TI - [Compared validity of the criteria of quantification of aortic insufficiency using pulsed and continuous Doppler]. AB - In order to evaluate the severity of aortic valve regurgitation (AVR) by means of simple criteria, we compared the feasibility and reliability of two methods: (1) pulsed doppler ultrasound suprasternal recording in the aortic sinus area, with calculation of the regurgitation fraction by planimetry of the systolic and diastolic curves, and with measurement of end-diastolic velocity, or end diastolic doppler effect (EDDE); this was done in 114 subjects (84 patients with AVR and 30 controls); (2) continuous wave doppler ultrasound apical recording of the left intraventricular jet, with measurement of the velocity decrease slope (S) and of the velocity half-decrease time (T 1/2); this was done in 46 patients with AVR. Doppler results were compared with Seller's angiographic classification of AVR in 4 grades. Planimetry could be performed in only 41% of patients in this series. This measurement seems to be feasible only when perfect recording of an increased systolic flow (peak velocity higher than 1.2 m/s) can be performed, which is usually limited to cases with major regurgitation. EDDE was easier to record (84/84 patients). When above 5 cm/s it is a good reflection of AVR severity, and when above 20 cm/s it indicates a major AVR (3/4 or 4/4 at angiography), with an 81% sensitivity and a 91% specificity. Continuous wave doppler ultrasound apical recording could be used in 80% of the cases (37/46 patients). With this method, a more than 3 m/s slope is a highly specific (8/8) but not very sensitive (8/13) sign of major AVR. A T 1/2 value lower than 650 ms is a specific (12/12) and sensitive (12/13) sign of severe AVR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973295 TI - [Role of atrial natriuretic peptide in the renal response to acute volume expansion in the normal and essential hypertensive patient]. AB - An exaggerated natriuretic response to volume expansion (VE) is observed in many essential hypertensive patients. The plasma levels of atrial natriuretic peptide (ANP) were measured in 11 normal subjects (NT) and 12 patients with mild essential hypertension (EH) during VE (1 800 ml isotonic saline IV over 3 hours). NT and EH groups were similar with respect to age and basal levels of renin, aldosterone and ANP (34.5 +/- 5.5 in NT and 32.5 +/- 6.3 pg/ml in EH, mean +/- sem). In response to VE, ANP increased to the same extent in both groups (a change of + 19.3 +/- 5.2 in NT and 22.2 +/- 7.1 pg/ml in EH) despite the marked difference in observed natriuresis (36 +/- 3.5 in NT and 54.9 +/- 6.3 mmol/3 in EH, p less than 0.02). The change in ANP induced by VE was inversely correlated with the fall in hematocrit and the variation in fractional excretion of sodium in both groups. These results suggest that atrial natriuretic peptide may participate in the control of the renal response to isotonic volume expansion, but they do not support a predominant role of atrial natriuretic peptide in the exaggerated natriuretic responses to volume expansion of patients with essential hypertension. PMID- 2973296 TI - [Arterial hypertension with hyperkalemia, tubular acidosis and normal renal function: Gordon syndrome and/or pseudohypoaldosteronism type II?]. AB - Basic examination of Mr S., 45 years of age, short in stature and overweight (1.60 m, 76 kg), was carried out because of the mild hypertension (mean AP 125 mm Hg) from which he had suffered for 20 years. The results were as follows: (1) variable hyperkalemia: plasma potassium values were 5.3 to 6.9 mmol/l; (2) normal renal function: serum creatinine 91.5 mumol/l, clearance of inulin 136.6 ml/mn; (3) proximal tubular acidosis: plasma bicarbonate and chloride values were 18.4 and 109 mmol/l, respectively; urinary pH was 7.1 with negative H+ ions urinary excretion (-33 mumol/mn); when plasma bicarbonate level was raised to 26 mmol/l by acute loading, fractional excretion of bicarbonate increased to 19,5 p. 100 while plasma potassium value decreased to 4.2 mmol/l; (4) low PRA (0.29 ng/ml/h) and normal plasma aldosterone concentration (63 pg/ml) with a normal intake of sodium and in a recumbent position. Plasma atrial natriuretic factor (ANF) level was normal: 14 fmol/ml. Intravenous infusion of ANF for 2 h (1 microgram/mn) induced the expected increases in urinary flow rate, and sodium and potassium excretions (+226, +307 and +171 p. 100, respectively). Intravenous infusion of isotonic saline (2 l in 2 h) and oral administration of fludrocortisone acetate for 4 weeks (400 micrograms per day) resulted in a normal decrease in PRA and plasma aldosterone concentration, a normal rise in plasma ANF level (22 and 42 fmol/ml) while slightly increasing AP without improving bicarbonaturia and acidosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973297 TI - [Effects of ventricular hypertrophy following experimental renovascular hypertension on the elasticity of the papillary muscle]. AB - A precise method was developed for detecting muscular stiffness changes in the left ventricular hypertrophy by chronic pressure overload in rats (2 kidney-1 clip renal hypertensive rats). The technique of Quick-Release allowed the measurement of the "stress-strain" relation (delta-epsilon), the elastic stiffness (d delta/d epsilon) and the stiffness constant (K) in isolated papillary muscle from normal and hypertrophied hearts at rest and at the maximal time course of activation during isometric contraction. (Table: see text). The results of this study reported that the cardiac hypertrophy induced by experimental pressure overload was not associated with significant alteration of myocardial stiffness. So, it would seem that a muscular stiffness modification was unresponsible for the ventricular compliance change of clinical and experimental results. PMID- 2973298 TI - [Treatment of renovascular arterial hypertension: angioplasty versus surgery]. AB - Percutaneous transluminal angioplasty is being more and more performed as the first choice treatment of renovascular hypertension. However, very few studies are devoted to the comparison of angioplasty versus surgery in treating renovascular hypertension and no one is prospective. A group of 25 patients with renovascular hypertension who underwent surgical treatment in the years 78-82 was compared to a group of 32 patients who underwent transluminal angioplasty in the years 82-86. Age, sex, etiology and severity of the arterial stenosis, blood pressure before treatment were similar in both groups. Average follow up was 44 +/- 33 months in the surgical group (S), 18 +/- 15 months in the angioplasty group (A). In the surgical group, we noticed one death, 20 p. 100 of major complications (thrombosis or stenosis of bypass grafts). In the angioplasty group, there was no death, 82 p. 100 of initial success, 12 p. 100 of major complications which needed surgery, 22 p. 100 of restenosis which were treated with a second angioplasty. Patients were classified as cured (BP less than 140/90 without treatment and perfect renal vascularisation) improved (BP less than 160/100 without treatment or with treatment if reduced and renal artery stenosis less than 50 p. 100), unchanged (BP greater than 160/100 and/or renal artery stenosis greater than 50 p. 100 or thrombosis). (Table: see text). Results are similar in both groups. The simplicity of the procedure, a lower cost and the absence of lethal complications are in favour of the transluminal angioplasty as the first choice treatment of renovascular hypertension. PMID- 2973299 TI - [Transluminal angioplasty in renovascular hypertension with renal insufficiency]. AB - From 1985 to 1986, 71 patients with renovascular hypertension were treated by percutaneous transluminal angioplasty (PTA). Among them, 13 (mean age 67 +/- 7 years) had a hypertension refractory to drug therapy (blood pressure: 194 +/- 33/103 +/- 15 mmHg) associated with an impaired renal function (creatinine clearance: 33 +/- 20 ml/mn). Ten had a stenosis in a solitary functioning kidney and a contralateral renal artery thrombosis. Three had bilateral renovascular stenosis. All patients had severe diffuse atherosclerotic disease, i.e. coronary heart disease (n = 7), carotid artery stenosis (n = 6), abdominal aortic aneurysm (n = 3) or arteritis (n = 5). Among these 13 patients, PTA could not be performed in one patient (failure to catheterize the stenosis) and two immediate renal artery dissections were observed: the first was accompanied by a thrombosis of the renal artery which could be successfully treated in emergency by surgical revascularization. The second occurred in a segmental renal branch and did not require surgery since it did not induced further impairment of renal function. Among the 10 remaining patients, nine PAT were classified as immediate angiographic success. One incomplete result required a second PTA 6 months later. Three important inguinal hematomas were observed and blood transfusion was required in 2. Seven patients have been reevaluated after a follow-up of 3 to 22 months. Restenosis occurred in two patients, 6 and 20 months respectively after PTA. A successful surgical revascularization was performed in these 2 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973300 TI - [Advantages of immediate coronary dilatation after thrombolysis using tissue-type plasminogen activator in acute myocardial infarction. A European Cooperative Group trial]. AB - A randomized trial was carried out in 367 patients with acute myocardial infarction to find out whether an "invasive" treatment consisting of intravenous thrombolysis with the tissue plasminogen activator (rt-PA) immediately followed by transluminal coronary angioplasty (group 1, n = 183) was superior to thrombolysis alone (group 2, n = 184). All patients received: 1. rt-PA (100 mg over 3 hours) started 156 minutes (range 30-294 mn) after the onset of symptoms; 2. heparin (5.000 IU as bolus injection, then 1.000 IU/hour), and 3. aspirin (250 mg i.v.). Coronary arteriography was performed 42 minutes (range 6-165 mn) later in 180 of the 183 patients in group 1. Following angioplasty, carried out in 168 patients, the immediate coronary patency rate was 89 p. 100 with a less than 50 p. 100 residual stenosis in 61 p. 100 of the patients. Immediate re-occlusion occurred in 23 p. 100 of the cases. Group 2 patients had a more favourable clinical course: mortality rate on the 14th day 3 p. 100 vs 7 p. 100, recurrent ischaemia within the first 24 hours 3 p. 100 vs 17 p. 100, and from 24 hours to the 14th day 11 p. 100 vs 13 p. 100, ventricular fibrillation 3 p. 100 vs 11 p. 100, haemorrhagic complications 23 p. 100 vs 41 p. 100.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973301 TI - [Effects of nicardipine on left ventricular hypertrophy of the rat with renovascular arterial hypertension]. AB - This study investigate the effects of Nicardipine treatment on regression of left ventricular hypertrophy (LVH), coronary hemodynamic and myocardial mechanical performance. 30 Sprague-Dawley male rats were divided into 3 groups: sham operated rats control group (SHC), untreated hypertensive rats group (RHR-U), treated hypertensive rats group (RHR-N). Systemic and coronary hemodynamics were determined by using left atrial injection of radioactive microspheres, 16 weeks after clipping. Mechanical performance was measured on isolated papillary muscle from the same animal. Results (mean +/- SEM) (Table: see text). Nicardipine treatment (10 to 15 mg intraperitoneal dosage during 8 weeks), led to: an efficient but incomplete control of hypertension. a reduction of left ventricular mass in proportion lesser than pressure decrease. a raise of coronary blood flow at rest with inversion of flow distribution between endocardium an epicardium. a decrease of "maximal" coronary blood flow. a reversal of impaired myocardial mechanical parameters towards control values except for contraction timing parameters. Decrease of "maximal" coronary blood flow could have deleterious effects on cardiac function. PMID- 2973302 TI - An overview of the thrombolysis and angioplasty in acute myocardial infarction (TAMI) trials. PMID- 2973303 TI - Energy cost of ambulation in healthy and disabled Filipino children. AB - The energy expenditures (Ee) for locomotion by nondisabled and disabled Filipino children aged 7 to 13 were determined and compared using indirect calorimetry. Forty-one controls (20 boys and 21 girls) ambulated at a comfortable pace; 16 children (eight boys and eight girls) with lower extremity poliomyelitis of varying severity ambulated by (1) wheelchair propulsion, (2) bilateral axillary crutches, (3) unilateral lower extremity ankle-foot orthoses or knee-ankle-foot orthoses, and (4) unassisted. Disabled children, regardless of their mode of ambulation, had to expend significantly more energy to ambulate than normal children (p less than 0.05). Wheelchair propulsion cost 16% more energy than the normal gait; crutch ambulation cost 41% more than the control. Children using unilateral braces sacrificed speed to attain near-normal Ee. When they ambulated without orthoses, their Ee increased by 109% over the control. In ascending order, the least energy was expanded by normal ambulation followed by disabled ambulation with unilateral brace, disabled propelling a wheelchair, disabled ambulation with bilateral axillary crutches, and disabled ambulation without brace. Efficiency of locomotion was reflected in the values obtained for Ee in terms of kcal x 10(-3)/kg/m, as demonstrated by the lower Ee but slower ambulation of children with braces, as compared to the nondisabled children. PMID- 2973304 TI - [The chronic form of the Budd-Chiari syndrome]. AB - A rare chronic course of Budd-Chiari syndrome associated with thrombosis of the portal vein was observed in a 30-year-old male patient suffering from postmyocarditic cardiosclerosis. At the age of 24 the patient had infectious allergic myocarditis, was hospitalized and rehospitalized for circulatory insufficiency. Upon 3 years since the disease onset the patient was admitted to a hematological department for progressive enlargement of the spleen. The diagnosis on discharge was idiopathic myelofibrosis with portal hypertension. The treatment included prednisolone, blood transfusions, myelosan. In 1987 the patient presented with enlarged liver and spleen, ascites, gastric and esophageal varicosis, augmenting hepatic insufficiency clinically evaluated as hepatic cirrhosis. Postmortem examination revealed macrofocal cardiosclerosis, splenomegaly, ascites, portal varicosis, enlarged nutmeg liver with smooth surface. Microscopically there was phlebosclerosis and phlebothrombosis varying in duration and involving predominantly medial branches of the hepatic and portal veins, liver fibrosis. The findings provided evidence for the final diagnosis of Budd-Chiari syndrome running an uncommon chronic course. PMID- 2973305 TI - [Recollections of work in Orenburg during World War II (1941-1944)]. PMID- 2973306 TI - District leaders meet: treat AIDS victims, they say. PMID- 2973307 TI - Allergic contact dermatitis to mace tear gas. PMID- 2973308 TI - Iodic eruptions. PMID- 2973310 TI - [Acinetobacter calcoaceticus var. Iwoffi as the cause of abscess formation in Han:NMRI nude mice]. PMID- 2973309 TI - 2-Mercaptopropionylglycine improves aortic flow after reoxygenation in working rat hearts. AB - The cardioprotective concentration range of the thiol drug 2 mercaptopropionylglycine (MPG) was investigated during reoxygenation after 30 min of hypoxia. It was found that aortic flow and frequency were increased by 1 mM MPG. Coronary flow, systolic and diastolic pressure were not significantly influenced by the drug. Mitochondria, isolated from hearts after termination of the perfusion phases, revealed increased values of RCI, when MPG had been present in the previous reoxygenation phase at 1 mM concentration. 5 mM MPG no longer showed a protective influence on the above cardiac and mitochondrial parameters. ATPase activities were decreased at 1 mM MPG by 14% and at 5 mM MPG by 40% of the controls. The latter concentration of MPG also doubled the inhibitory action of carboxyatractyloside on ATPase activity, indicating a structural change of the adenine nucleotide carrier. 1 mM MPG is considered an optimal therapeutic range in this model. The mechanism of action most probably includes an SH/-S-S interchange reaction as well as a free radical scavenging mechanism. For many thiols, the latter is known to occur in the presence of metal ions. PMID- 2973311 TI - GM2-gangliosidosis B1 variant: a wide geographic and ethnic distribution of the specific beta-hexosaminidase alpha chain mutation originally identified in a Puerto Rican patient. AB - A point mutation within exon 5 of beta-hexosaminidase alpha chain gene was identified earlier in a Puerto Rican patient with GM2-gangliosidosis B1 variant (the DN-allele) [K. Ohno and K. Suzuki: J. Neurochem. 50:316-318, 1988]. Oligonucleotide probes designed to detect either the normal or the DN-allele showed that four additional patients carried the same mutation. These patients were of Italian, French, Spanish, and English/Italian/Hungarian origin. Three of them, as well as our original patient, were compound heterozygotes with positive signals for both the mutant and normal probes, while the Spanish patient was positive only for the DN-allele. A patient from Czechoslovakia was negative for the DN-allele. Thus, the specific mutation originally found in the Puerto Rican patient has a surprisingly wide geographic and ethnic distribution. This mutation can account for the B1 variant phenotype in five of the six B1 variant patients so far examined. PMID- 2973313 TI - Cloning of a complete cDNA encoding human aromatase: immunochemical identification and sequence analysis. AB - A complete cDNA clone encoding a human aromatase was isolated from a human placental cDNA library in lambda gt11. An antibody to the polypeptide specified by the isolated clone was prepared, and Western blot analysis and antibody inhibition experiments of human placental aromatase activity confirmed the identification of the clone as aromatase cDNA. The isolated aromatase cDNA clone of 3030 bp with two unique EcoRI sites contained a 3'-noncoding region of 1397 bp, an open reading frame of 1509 bp encoding 503 amino acid residues, and a 5' noncoding region of 124 bp. Analysis of the amino acid sequence of aromatase and comparison of aromatase with other forms of cytochrome P-450 indicated that this enzyme is a unique form of the cytochrome P-450 superfamily. PMID- 2973312 TI - Atrial natriuretic peptide gene expression and its secretion by pneumocytes derived from neonatal rat lungs. AB - Using a primary culture of pneumocytes derived from neonatal rat lungs, we investigated the synthesis and secretion at transcriptional and peptide levels of pulmonary rat(r) atrial natriuretic peptide (ANP). Total RNA extracted from pneumocytes contained a hybridizing RNA band of the same size as atrial rANP mRNA. Immunoreactive (IR)-rANP content in pneumocytes was 0.5% of that in atrial myocytes, and 8.6% of that in ventricular myocytes, while the secretory rate from pneumocytes was about 7% of atrial and ventricular myocytes. Triiodothyronine (T3, 5 x 10(-10) to 5 x 10(-8) M), dexamethasone and testosterone (5 x 10(-9) to 5 x 10(-8) M) significantly stimulated the synthesis of IR-rANP by pneumocytes in a dose-dependent manner. However, the stimulatory effect exerted by T3 on rANP synthesis, unlike in the case of cardiocytes, was much more potent than that of dexamethasone, as evidenced by the significant difference in potency at both transcriptional and peptide levels. The present study suggests that ANP secreted from lungs may at least in part contribute to circulating ANP pool, and that the tissue-dependent difference of sensitivity to thyroid hormone may play an important role in the regulation of developmental ANP gene expression in mammalian lungs. PMID- 2973314 TI - Calcium-stimulated ATPase activity in plasma membrane vesicles from pancreatic acinar cells. AB - Plasma membrane vesicles were prepared from rat pancreatic acinar cells using nitrogen cavitation and magnesium precipitation. The vesicles exhibited ATPase activity that was stimulated by submicromolar concentrations of free calcium and was dependent upon the presence of magnesium. This enzyme activity was localized to the cytoplasmic surface of the plasma membrane by two criteria. First, no activity was observed when intact cells replaced the membrane vesicles in the assay. Second, right side-out and inside-out vesicles were separated using concanavalin A sepharose-B. The calcium-stimulated, magnesium-dependent ATPase activity per mg protein in the inside-out fraction was 60% greater than that occurring in the mixed vesicle preparation. These results indicate that the plasma membrane of pancreatic acinar cells has a calcium-stimulated, magnesium dependent ATPase located on its cytoplasmic surface and that this enzyme is stimulated by submicromolar concentrations of free calcium. PMID- 2973315 TI - Rat liver carboxylesterase: cDNA cloning, sequencing, and evidence for a multigene family. AB - A cDNA clone was isolated from a rat liver lambda gt11 expression library by screening with polyclonal antibodies raised against a rat liver microsomal carboxylesterase. This clone of 1.8 kb contained an open reading frame encoding a mature protein of 531 amino acids with a predicted molecular weight of 58,084. The 5' portion of the clone coded for 9 amino acids of a putative signal peptide. The 3' end of the clone included an untranslated region and a poly (A) tail. Carboxylesterase active site regions, five potential N-linked glycosylation sites, and 2 postulated cystine disulfide bridges were found in the cDNA-deduced amino acid sequence. Sequences obtained from tryptic peptides and the NH2 terminus of the purified native carboxylesterase were aligned with the deduced amino acid sequence, and the overall identity was 84%. Southern blot analysis suggested the presence of multiple genes. Thus it is concluded that we have cloned a rat liver carboxylesterase, and that this enzyme is a member of a multigene family. PMID- 2973316 TI - Sucrose feeding prevents changes in myosin isoenzymes and sarcoplasmic reticulum Ca2+-pump ATPase in pressure-loaded rat heart. AB - Pressure-overload due to banding of the abdominal aorta in rats for 10 weeks resulted in cardiac hypertrophy, redistribution of myosin isoenzymes and reduction in the sarcoplasmic reticulum (SR) Ca2+-stimulated ATPase activity. Administration of sucrose in the drinking water (0.8%, w/v) to rats prevented changes in myosin isoenzymes and SR Ca2+-stimulated ATPase in hypertrophied hearts. This beneficial effect of sucrose feeding with respect to remodeling of the subcellular organelles in the myocardium was not associated with any significant changes in plasma glucose or thyroid hormone levels. It is suggested that the prevention of subcellular changes in the hypertrophied hearts due to sucrose feeding may be due to a shift in fuel utilization by the myocardium. PMID- 2973317 TI - Changes in intracellular redox and energy status during induced transplasma membrane electron transport in Cuscuta protoplasts. AB - Extracellular reduction of ferricyanide was exhibited by isolated Cuscuta protoplasts. A larger decrease in NADH than NADPH levels of the ferricyanide treated protoplasts pointed to the major involvement of the former as an electron donor. Glutathione levels were also found to be lowered in similarly treated tissue. The time-dependent variation in intracellular ATP levels in presence of ferricyanide supported the concept of plasma membrane ATPase activation during transplasma membrane electron transport in eukaryotes. PMID- 2973318 TI - Effects of atrial natriuretic factor on urinary concentration of catecholamines and renin secretion in dogs. AB - This study evaluated the effects of synthetic atrial natriuretic factor (ANF) on renal hemodynamics, urinary excretion of electrolytes, norepinephrine (NE), and dopamine (DA); and renal production of renin in anesthetized dogs. Following a bolus (1 micrograms/kg body weight) and infusion (0.1 microgram/kg/min) for 30 min, there was significant increase in urine flow (220 +/- 41%), glomerular filtration rate (72 +/- 14%), and urinary sodium excretion (170 +/- 34%). There was a decrease in renin secretory rate and the concentration ratio of urine NE to DA following ANF was decreased (p less than 0.05). These data suggest that ANF decreases renal production of NE and renin. PMID- 2973319 TI - Detection of an ATPase activity in rat liver peroxisomes. AB - An ATPase co-sedimenting with rat liver peroxisomes has been detected after subcellular fractionation. The activity is Mg2+ dependent, with pH optimum of 7.5 and is inhibited by NEM and DCCD but not by oligomycin. Partial inhibition of the mitochondrial ATPase allows to detect the peroxisomal activity in the gradients. Protease inactivation and solubilization data suggests that the activity resides in a protein of the peroxisomal membrane, exposed to the cytosol. PMID- 2973320 TI - Transfer of N-acetylglucosamine to endogenous glycoproteins in the nucleus and in non-nuclear membranes of rat hepatocytes: electrophoretic analysis of the endogenous acceptors. AB - The transfer of N-acetyl(14C)glucosamine from UDP-N-acetyl(14C)glucosamine to endogenous glycoproteins acceptors were studied comparatively in the nuclei and in the non-nuclear membranes of rat hepatocytes. Electrophoretic and autoradiographic analysis show that most of the glycoprotein acceptors of the nuclei differ from those of the non-nuclear membranes in terms of molecular weight. In addition, it may interesting to mention that in the nuclear fraction a 30% inhibition by tunicamycin is obtained for concentrations as low as 0.03 microM, whereas at this concentration no effect is detected in the non-nuclear membranes. In the presence of 0.2 microM tunicamycin, the inhibition does not go beyond 25% in the latter fraction but goes up to 80% in the former. The previous results demonstrate clearly that a particular glycosylation reaction occurs in the nucleus. PMID- 2973321 TI - Degradation of atrial natriuretic peptides by an enzyme in rat kidney resembling neutral endopeptidase 24.11. AB - The inactivation of rat atrial natriuretic factor (ANF) was studied using a bioassay, ANF-stimulated guanylate cyclase activity. Rat kidney membranes degraded ANF into biologically inactive forms. The primary cleavage site appears to be the Cys105-Phe106 bond. The degradation, measured by HPLC, followed classical Michaelis-Menten kinetics. The sensitivity of the enzyme to inhibitors suggested it to be a metalloendopeptidase, resembling neutral endopeptidase 24.11. When this enzyme, characterised by its enkephalin-degrading activity, was compared to the enzyme responsible for ANF inactivation, striking differences were found in tissue distribution, pH-dependence and sensitivity to protein modifying reagents. It is concluded that an enzyme similar to endopeptidase 24.11 may be responsible for inactivation of atrial peptides in the rat. PMID- 2973322 TI - Characterization of thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptors of rat platelets and their interaction with TXA2/PGH2 receptor antagonists. AB - Characterization of thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptors of rat platelets was performed on both intact platelets and crude membrane fractions. The binding of [3H]U46619, a stable TXA2 mimetic, to intact platelets was found to be saturable and displaceable. Scatchard analysis of equilibrium binding at 24 degrees revealed a single class of binding sites with a Kd of 37 nM and a Bmax of 160 fmol/10(8) platelets. The binding affinity of [3H]U46619 to the platelet membrane fractions was remarkably and specifically enhanced by addition of Mg2+ without alteration of the maximum density level. Kinetic analysis for [3H]U46619 binding to the membrane fractions in the presence of 20 mM MgCl2 gave a K1 of 6.9 x 10(6) M-1 min-1 and a K-1 of 0.25 min-1, yielding a Kd (K-1/K1) of 36 nM; the value corresponded well to Kd values from Scatchard analysis in both intact (37 nM) and crude membrane fractions (39 nM). A series of TXA2/PGH2 receptor antagonists completely suppressed U46619 binding to rat platelets as well as collagen-induced platelet aggregation. The rank order of binding affinities to rat platelets (intact platelets or crude membranes) among the respective antagonists correlated well with (a) that of human platelet membrane fraction and (b) the potencies for suppression of collagen-induced platelet aggregation in rat. These results may support our proposed mechanism of TXA2/PGH2 action in collagen-stimulated platelets [K. Hanasaki et al., Thromb. Res. 46, 425 (1987)] and also suggest that they may provide a simple technique for evaluating synthetic TXA2/PGH2 receptor antagonists. PMID- 2973323 TI - Inhibition of intracellular esterases by antitumour chloroethylnitrosoureas. Measurement by flow cytometry and correlation with molecular carbamoylation activity. AB - Antitumour chloroethylnitrosoureas (Cnus) decompose in physiological conditions yielding alkylating species and organic isocyanates. While antitumour activity is mainly attributed to the alkylation of DNA, carbamoylation of intracellular proteins by isocyanates may also have pharmacological and toxicological relevance. We previously reported a novel dynamic flow cytoenzymological assay for esterase inhibition in intact murine cells by BCNU and related isocyanates, and proposed that this might form the basis of an assay for intracellular carbamoylation. We have now examined a wide range of Cnus, isocyanates, and alkylating agents for their ability to inhibit cellular esterases. BCNU, CCNU, their derived isocyanates, and the 4-OH metabolites of CCNU exhibited potent inhibitory activity (I50 values 5.5 x 10(-5)-7.3 x 10(-4) M). Chlorozotocin and GANU were relatively inactive (I50 much greater than 10(-2) M). ACNU, TCNU and the 2-OH metabolites of CCNU exhibited intermediate activity (I50 values 1.1 x 10(-3)-2.3 x 10(-2) M). Compounds not able to form isocyanates were essentially inactive. Poor membrane permeability was also implicated in the weak activity of chlorozotocin and GANU. There was overall a good correlation between esterase inhibition and chemical carbamoylating activity, but some particular differences were identified. We concluded that flow cytoenzymological assay appears to have the potential to provide useful measurement of intracellular protein carbamoylation by existing Cnus and novel derivatives, and also offers the advantage of cell subpopulation identification for in vivo evaluation of these agents. PMID- 2973324 TI - Effect of atrial natriuretic factor on brush border membrane transport of phosphate in phosphate-deprived rats. PMID- 2973325 TI - Ionophoretic and inhibitory action of the analgesic, diflunisal, on sarcoplasmic reticulum. AB - Diflunisal decreased the ATP-dependent transport rate and calcium accumulation by the sarcoplasmic reticulum. Inhibition of calcium transport by diflunisal was pH dependent, and a pKa of 6.7 to 6.9 was observed for the carboxylic acid group. In sealed sarcoplasmic reticulum vesicles, diflunisal at concentrations below 1 mM increased the rate of Ca2+-dependent hydrolysis of ATP; above 1 mM, the Ca2+ ATPase activity was inhibited. In purified Ca2+-ATPase, diflunisal acted only as an inhibitor. Methylation of the phenolic group of diflunisal eliminated both its analgesic and ionophoretic properties. PMID- 2973326 TI - [Atrial natriuretic peptides. I. Synthesis of C-terminal fragments]. AB - C-terminal fragments of atrial natriuretic peptides have been synthesized by classical methods of peptide chemistry in solution and characterized by various physico-chemical methods. The choice of the scheme and methods of synthesis is discussed. PMID- 2973327 TI - [Atrial natriuretic peptides. II. Synthesis of N-terminal fragments]. AB - N-terminal fragments of atrial natriuretic peptides have been synthetized by classical methods of peptide chemistry in solution and characterized by various physicochemical methods. The choice of the scheme and methods of synthesis is discussed. PMID- 2973328 TI - [Atrial natriuretic peptides. III. Synthesis and biological activity of alpha-r ANP, APII, APIII and des-Ser5,Ser6-APII]. AB - A series of atrial natriuretic peptides, viz., alpha-r-ANF, APII, APIII, and des Ser5, Ser6-APII, have been obtained by condensation of earlier synthesized fragments promoted with complex F. Acetylaminomethyl protecting group were removed from cystein residues with simultaneous cyclization of the peptide. Biological activity of the atrial natriuretic peptides was studied. PMID- 2973330 TI - Ultrastructural effects of diphosphonates on dental enamel. PMID- 2973329 TI - Calcium transport by a calmodulin-regulated Ca-ATPase in the enamel organ. PMID- 2973331 TI - The atrial peptide system in cardiac disease. AB - Atrial natriuretic factor (ANF) is a peptide hormone of cardiac origin, released in response to atrial distention, that increases sodium excretion, inhibits the renin-angiotensin-aldosterone (RAA) system, and decreases arterial pressure. Congestive heart failure (CHF) is a clinical syndrome that is characterized by chronic atrial distention, sodium retention with edema, and activation of the RAA system. The role of ANF in the pathophysiology of CHF remains to be defined. Circulating ANF is greatly increased in congestive heart failure. The mechanism of this elevation of ANF is explained by increased synthesis and release of ANF from the atria with atrial depletion of ANF in response to chronic atrial overload. Recent work also suggests the presence of ANF in ventricular myocardium in heart failure in which the ventricle is recruited to synthesize and release this peptide hormone. Despite increased circulating ANF in heart failure, the kidney retains sodium and is hyporesponsive to exogenous administration of ANF. The mechanism of this hyporesponsiveness is multifactorial but may be explained in part by activation of the intrarenal RAA system as well as by a reduction in renal perfusion pressure. A therapeutic role for ANF remains unclear. To date, studies conflict, but some investigations of ANF infusions in man support a unique and selective renal vasodilator action in association with an ability to inhibit the RAA system. PMID- 2973332 TI - Aortic stenosis produces hypertrophy of the rat heart without causing an accumulation of N1-acetylspermidine. AB - Isoprenaline treatment causes cardiac hypertrophy and an accumulation of N1 acetylspermidine in the rat heart. To determine whether the cardiac hypertrophy is the cause of the increase in N1-acetylspermidine, we produced cardiac hypertrophy by constriction of the aorta and analyzed polyamines in the hearts of these rats 1, 3, and 10 days after the aortic constriction. Our results show that compared to sham-operated animals, this treatment caused a 60% increase in putrescine and a 30% increase in spermidine by day 10, but not the expected increase in N1-acetylspermidine. We conclude that N1-acetylspermidine is not induced by a cardiac overload. PMID- 2973333 TI - Binding of a 30-kDa protein to the pyruvate kinase gene of Neurospora crassa. AB - Extracts of a wild-type strain of Neurospora crassa, electrophoresed on SDS polyacrylamide gels and electroblotted onto nitrocellulose sheets, were hybridized to an end-labelled pyruvate kinase (PK) gene fragment containing the 5' noncoding sequence and a large part of the coding region. A 30-kDa protein was found to bind strongly to the PK gene DNA, while binding weakly to plasmid pUC12 DNA and to total N. crassa DNA. Probing of blots with individual restriction fragments derived from the PK gene showed that the protein binding occurred primarily to the 5' noncoding region. Nonspecific DNA from pUC12, PK gene DNA from the recombinant plasmid pNP460 (pUC12 containing a 1.8-kilobase EcoRI insert of the PK gene DNA), along with a 0.7-kilobase EcoRI-AccI restriction fragment containing the 5' flanking region, were used in filter-binding experiments to analyze the kinetics of binding. Formation of protein-DNA complexes was demonstrated by monitoring the electrophoretic mobility of this fragment on nondenaturing gels. PMID- 2973334 TI - [Behavior of beta-endorphin, GH, cortisol and glucagon during insulin hypoglycemia in type II diabetes]. PMID- 2973335 TI - [Endocrine aspects of obesity: evaluation of beta-endorphin during insulin hypoglycemia]. PMID- 2973336 TI - [Transmembrane transport of calcium in normal and pathological pregnancy. 1st experimental data]. PMID- 2973337 TI - Bone formation in experimental myositis ossificans. Light and electron microscopy study. AB - The development of ectopic ossification in experimental myositis ossificans of the rabbit thigh was studied. The right hind limb of 25 rabbits was immobilized with the knee in extension. Once a day the limb was passively mobilized for 2-3 minutes. The animals were killed 3, 5, 7, 14, 21, 28 and 35 days after the beginning of the experiment. Specimens for light and electron microscopy were obtained from both hind limbs. Extensive necrosis and fibrosis were observed in the right vastus intermedius muscle during the first week. Proliferation of chondrocytes and osteoblasts with newly formed woven bone and cartilage formation were found in the periosteum within 7 and 14 days after the beginning of the experiment. Intensive enchondral ossification and hard calcified tissue were observed later. It was evident that bone formation in this experimental model started in the periosteum after necrosis of the adjacent skeletal muscle. Therefore the temporal and spatial relationship of traumatic changes in the periosteum and muscle seems important for the development of myositis ossificans. PMID- 2973338 TI - Galactosylation of endogenous proteins from human platelets. AB - Human platelets have been shown to contain the enzyme glycoprotein:galactosyltransferase that catalyzes the transfer of galactose to an endogenous protein acceptor present in the platelet. Galactosylation of added ovalbumin also occurs. The activity was extracted with 30 mM Tris buffer (pH 7.5). The endogenous activity was enriched 1.4-fold (compared with the crude homogenate) in the fraction, 105,000 g pellet, and the exogenous enzyme was retained in the respective supernatant. The two galactosyltransferase activities showed proportionality to time, protein, and substrate concentration, and were identical in pH dependence and Mn+2 requirement. The effect of Triton X-100 (range 0-1.5%) in the assay system appeared to be different for both activities: with the optimum concentration of detergent (0.15%) the endogenous activity increased by 50% whereas the exogenous activity was augmented 5-fold. From a number of sugar nucleotides tested as glycosyl donor into the endogenous proteins, the optimum substrate was UDP-Glc (100%), followed by UDP-Gal (80%), GDP-Man (24%), UDP-Glc-NAc (21%), UDP-Xyl (19%), and ADP-Glc (5%). An appropriate exogenous acceptor for UDP-Glc as donor was not found. The different solubilization of galactosyl- and glucosyltransferase activities by Triton X-100 suggests that they are distinct enzymes. In addition, the exogenous galactosyltransferase activity achieved after the treatment was much higher (940%) than the endogenous (26%). It is suggested that these differences on both galactosyltransferases could reflect changes in the accessibility of the exogenous substrate to the enzyme. PMID- 2973339 TI - Effect of contrast media on beta-endorphin secretion. An in vitro study. AB - The central nervous system may be highly susceptible to the toxic effects of contrast media (CM). Previous experiments demonstrated that vasopressin is released after the intravenous administration of CM. The present study examined the response of the opiocortin system to CM. Neurons of the rat basal hypothalamus, dispersed and attached to Cytodex-3 beads, were perfused with sodium diatrizoate, metrizamide or iohexol (3 mg iodine/ml). The effluent was collected, and the beta-endorphin (B-E) content was measured by a radioimmunoassay technique. Results, normalized to the internal positive control, were compared with release from normal saline (negative control) by analysis of variance. Diatrizoate and metrizamide caused significant release of B-E (p less than 0.03). Iohexol did not stimulate release of B-E. These results suggest that diatrizoate and metrizamide, but not iohexol, can stimulate the release of hormones from hypothalamic neurons. The phenomenon may play a role in some reactions to intravascular CM administration since these neurons are not protected by a blood-brain barrier. PMID- 2973341 TI - Predicted and measured oxygen concentrations in the circle system using low fresh gas flows with oxygen supplied by an oxygen concentrator. AB - An oxygen concentrator based on a zeolitic molecular sieve has been used to supply piped oxygen in a District General Hospital; such oxygen contains 5% argon. If concentrator oxygen is used in the closed circuit, argon accumulates. The present investigation defines the extent and time course of argon accumulation in partially and fully closed breathing systems. A theoretical model is presented which predicts the extent and rate of inert gas accumulation. The predictions have been tested in a volunteer under laboratory conditions and in five anaesthetized patients. It is recommended that concentrator oxygen may be used for low flow anaesthesia of indefinite duration, provided the fresh oxygen flow to the circuit is at least twice the oxygen consumption. If the circuit is totally closed, periodic opening of the circuit is necessary. PMID- 2973342 TI - Aseptic periprosthetic fluid collection: a late complication of Dacron arterial bypass. AB - Fluid collection encompassing an arterial prosthesis often leads one to suspect graft infection. Four cases of aseptic seroma occurring three to five years after insertion of knitted Dacron implants for bypass are reported. The collection was serohematic in three cases, puriform in the other. Repeated bacteriological investigations did not reveal any growth and laboratory tests did not demonstrate any signs of infection. The following pathophysiologic hypotheses have been advanced: modification of the permeability of the prosthetic wall, immunoallergic reaction to the graft, and aging hematoma. The role played by anticoagulant is debatable. Management by needle aspiration or drainage failed. Multiple recurrences led to prosthetic replacement in three cases. Treatment consists of replacing the graft by one of a different fabric. These collections must be distinguished from postoperative lymphoceles, anastomotic false aneurysms, hematomas and infections. PMID- 2973340 TI - Localisation of atrial natriuretic peptide immunoreactivity in the ventricular myocardium and conduction system of the human fetal and adult heart. AB - Atrial natriuretic peptide immunoreactivity was found in ventricular and atrial tissues with specific antisera raised to the amino and carboxy terminal regions of the precursor molecule. In 13 developing human hearts (7-24 weeks' gestation) the immunoreactivity was concentrated in the atrial myocardium and ventricular conduction system but it was also detected in the early fetal ventricular myocardium. Immunoreactivity in five normal adults was largely confined to the atrial myocardium although it was also found in the ventricular conduction tissues of hearts removed from 10 patients who were undergoing cardiac transplantation. The ventricular conduction system is an extra-atrial site for the synthesis of atrial natriuretic peptide. In the failing heart this synthesis may be further supplemented by expression of the gene in the ventricular myocardium. It is possible that ventricular production of the peptide contributes to the raised circulating concentrations of atrial natriuretic peptide immunoreactivity found in severe congestive heart disease, particularly in patients with dilated cardiomyopathy. PMID- 2973343 TI - Prospective randomized trial of polytetrafluoroethylene and Dacron aortic prosthesis. I. Perioperative results. AB - Over a two year period 80 patients were entered into a prospective randomized trial comparing polytetrafluoroethylene (PTFE) and Dacron infrarenal aortic reconstructions. Fifty-four patients were treated for aneurysm (30 single tubed grafts; 24 bifurcation grafts), and 26 patients were treated for occlusive disease (26 bifurcation grafts). The groups were matched for age, sex and preoperative risk factors. Five patients died after operation (6.3%) including two from hemorrhage, but there were no significant differences in mortality and morbidity between the PTFE and Dacron groups. The volume of blood lost at operation (1930 +/- 1340 ml, all patients); the volume of blood transfused (2.98 +/- 2.43 units); the volume of crystalloids infused (3050 +/- 1390 ml); the intraoperative heparin dosage (67.9 +/- 20.5 mg); the clamp time (71.6 +/- 34.5 min); and the total operating time (228.1 +/- 78.3 min) also showed no significant differences between PTFE and Dacron. The ankle systolic pressure index rose more for PTFE (0.96 +/- 0.24) than for Dacron (0.82 +/- 0.20; P less than 0.002) at the time of discharge. This partially reflects a difference in the index between the groups before operation (PTFE 0.79 +/- 0.30; Dacron 0.72 +/- 0.32), but it may also indicate that PTFE is less thrombogenic than Dacron. PMID- 2973344 TI - Adverse cutaneous drug reactions in AIDS. AB - A maculopapular rash is reported in a girl with AIDS and Pneumocystis carinii pneumonia treated with co-trimoxazole. AIDS patients seem to be at increased risk of adverse drug reactions. PMID- 2973345 TI - Terminal deoxynucleotidyl transferase and HLA-DR expression appear unrelated to prognosis of acute myeloid leukaemia. AB - Mononuclear cells from peripheral blood or bone marrow from 314 patients with acute myeloid leukaemia were examined for the presence of nuclear terminal deoxynucleotidyl transferase (304 patients), surface membrane expression of HLA DR (314 patients) and the common acute lymphoblastic leukaemia antigen (281 patients). All patients were treated with identical remission induction chemotherapy, and morphological diagnosis was carried out in a central laboratory. The overall complete remission rate was 70%. There were no significant correlations between the immunological markers and complete remission rate, duration of remission, or survival. PMID- 2973346 TI - Protein S and C4b-binding protein in fetal and neonatal blood. AB - The levels of protein C, free protein S, C4b-binding protein and the distribution of total protein S between a free form and a conjugated form with C4b-binding protein, were measured in fetuses between 15 and 35 weeks of gestation, in newborns and in maternal blood throughout pregnancy. Fetal and newborn umbilical blood samples were obtained by cordocentesis; in the case of newborns, immediately after delivery. C4b-binding protein was detected in five out of seven newborns and in only three out of 20 fetuses. The fetuses (21, 24, 29 weeks, respectively) whose blood contained C4b-binding protein (10%, 29%, 12% of adult level, respectively), did not survive: the possible cause of death for one was given as viral infection; all three were severely deformed. Only free protein S was detected in fetal blood and the mean value was 40%. By contrast, the mean levels of protein C in the fetuses increased from 11% to 15.5% of adult levels with advancing gestation. We conclude that the relatively high levels of free protein S in fetal blood may contribute to its non-coagulability and compensate for the low protein C levels. PMID- 2973347 TI - Roles of phosphorylation and nucleotide binding domains in calcium transport by sarcoplasmic reticulum adenosinetriphosphatase. AB - The roles of the phosphorylation (phosphorylated enzyme intermediate) and nucleotide binding domains in calcium transport were studied by comparing acetyl phosphate and ATP as substrates for the Ca2+-ATPase of sarcoplasmic reticulum vesicles. We found that the maximal level of phosphoenzyme obtained with either substrate is approximately 4 nmol/mg of protein, corresponding to the stoichiometry of catalytic sites in our preparation. The initial burst of phosphoenzyme formation observed in the transient state, following addition of either substrate, is accompanied by internalization of 2 mol of calcium per mole of phosphoenzyme. The internalized calcium is then translocated with a sequential pattern, independent of the substrate used. Following a rate-limiting step, the phosphoenzyme undergoes hydrolytic cleavage and proceeds to the steady-state activity which is soon "back inhibited" by the rise of Ca2+ concentration in the lumen of the vesicles. When the "back inhibition" is released by the addition of oxalate, substrate utilization and calcium transport occur with a ratio of 1:2, independent of the substrate and its concentration. When the nucleotide binding site is derivatized with FITP, the enzyme can still utilize acetyl phosphate (but not ATP) for calcium transport. No secondary activation of acetyl phosphate utilization by the FITC-enzyme was obtained with millimolar nucleotide. These observations demonstrate that the basic coupling mechanism of catalysis and calcium transport involves the phosphorylation and calcium binding domains, and not the nucleotide binding domain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973348 TI - Interaction of spin-labeled nucleotides with sarcoplasmic reticulum adenosinetriphosphatase. AB - Spin-labeled derivatives of AMP-PCP, ATP, and 2'-deoxy-ATP, with a nitroxide moiety attached to the ribose ring [3'-O-(1-oxy-2,2,5,5-tetramethylpyrroline-3 carbonyl)nucleotide], are used to study the nucleotide binding site stoichiometry of sarcoplasmic reticulum (SR) ATPase. With all derivatives, a maximal binding of 4.5 nmol/mg of SR protein is found, a value close to the number of phosphorylation sites obtained with ATP. The spin-labeled nucleotides cannot be utilized by the enzyme as substrates. Binding of spin-labeled nucleotides is inhibited by labeling the ATPase with fluorescein 5'-isothiocyanate, indicating that all the labeled nucleotide is located at the catalytic site. Additions of spin-labeled ATP to vesicle suspensions during steady turnover demonstrate competitive inhibition of both catalysis and the regulatory effect normally exhibited by ATP. As secondary binding of spin-labeled ATP is not detected at pertinent concentrations, it is suggested that both functions of ATP may be effected through a single site. PMID- 2973349 TI - Polypeptide domains of ADP-ribosyltransferase obtained by digestion with plasmin. AB - Proteolysis by plasmin inactivates bovine ADP-ribosyltransferase; therefore, enzymatic activity depends exclusively on the intact enzyme molecule. The transferase was hydrolyzed by plasmin to four major polypeptides, which were characterized by affinity chromatography and N-terminal sequencing. Based on the cDNA sequence for human ADP-ribosyltransferase enzyme [Uchida, K., Morita, T., Sato, T., Ogura, T., Yamashita, R., Noguchi, S., Suzuki, H., Nyunoya, H., Miwa, M., & Sugimura, T. (1987) Biochem. Biophys. Res. Commun. 148, 617-622], a polypeptide map of the bovine enzyme was constructed by superposing the experimentally determined N-terminal sequences of the isolated polypeptides on the human sequence deduced from its cDNA. Two polypeptides, the N-terminal peptide (Mr 29,000) and the polypeptide adjacent to it (Mr 36,000), exhibited binding affinities toward DNA, whereas the C-terminal peptide (Mr 56,000), which accounts for the rest of the transferase protein, bound to the benzamide Sepharose affinity matrix, indicating that it contains the NAD+-binding site. The fourth polypeptide (Mr 42,000) represents the C-terminal end of the larger C terminal fragment (Mr 56,000) and was formed by a single enzymatic cut by plasmin of the polypeptide of Mr 56,000. The polypeptide of Mr 42,000 still retained the NAD+-binding site. The plasmin-catalyzed cleavage of the polypeptide of Mr 56,000 42,000 was greatly accelerated by the specific ligand NAD+. Out of a total of 96 amino acid residues sequenced here, there were only 6 conservative replacements between human and bovine ADP-ribosyltransferase. PMID- 2973350 TI - Characterisation of a high affinity Ca2+-stimulated, Mg2+-dependent ATPase in the rat parotid plasma membrane. AB - Two Ca2+-stimulated ATPase activities have been identified in the plasma membrane of rat parotid: (a) a (Ca2+ + Mg2+)-ATPase with high affinity for free Ca2+ (apparent Km = 208 nM, Vmax = 188 nmol/min per mg) and requiring micromolar concentration of Mg2+ and (b) a (Ca2+ or Mg2+)-ATPase with relatively low affinity for free Ca2+ (K0.5 = 23 microM) or free Mg2+ (K0.5 = 26 microM). The low-affinity (Ca2+ or Mg2+)-ATPase can be maximally stimulated by Ca2+ alone or Mg2+ alone. The high-affinity (Ca2+ + Mg2+)-ATPase exhibits sigmoidal kinetics with respect to ATP concentration with K0.5 = 0.4 mM and a Hill coefficient of 1.91. It displays low substrate specificity with respect to nucleotide triphosphates. Although trifluoperazine inhibits the activity of the high affinity (Ca2+ + Mg2+)-ATPase only slightly, it inhibits the activity of the low affinity (Ca2+ or Mg2+)-ATPase quite potently with 22 microM trifluoperazine inhibiting the enzymic activity by 50%. Vanadate, inositol 1,4,5-trisphosphate, phosphatidylinositol 4,5-bisphosphate, Na+,K+ and ouabain had no effect on the activities of both ATPases. Calmodulin added to the plasma membranes does not stimulate the activities of both ATPases. The properties of the high-affinity (Ca2+ + Mg2+)-ATPase are distinctly different from those of the previously reported Ca2+-pump activity of the rat parotid plasma membrane. PMID- 2973351 TI - Purification and characterization of a Ca2+- or Mg2+-stimulated ATPase from plasma membrane enriched fractions of Dictyostelium discoideum. AB - Evidence is presented for the presence of both diethylstilbestrol (DES)-sensitive and DES-insensitive Mg2+-ATPase activities in plasma membrane enriched fractions of Dictyostelium discoideum. When removed from the membrane, the DES-sensitive activity is markedly less stable than the DES-insensitive activity, and the two activities display a number of quite distinct properties. The DES-sensitive enzyme has a decided preference for Mg2+ over Ca2+, displays saturation kinetics in response to ATP as substrate (Km = 0.2 mM) and has a narrow pH optimum range. In contrast, the DES-insensitive activity is stimulated equally by Mg2+ or Ca2+, is not saturable by ATP within the mM concentration range and has a much broader pH optimum. The DES-insensitive activity has been purified extensively. The purified enzyme is inhibited by vanadate and fluoride, but is insensitive to N,N' dicyclohexylcarbodiimide (DCCD), N-ethylmaleimide and thimerosal. In the absence of divalent cations, the enzyme displays a sigmoidal activity curve in response to substrate concentration, which is abolished by addition of either Mg2+ or Ca2+, suggesting a binding site for a divalent cation and a positive cooperative interaction. The enzyme is capable of hydrolyzing other nucleotide triphosphates and ADP, but is without activity on AMP, p-nitrophenyl phosphate and pyrophosphate. The enzyme has an apparent molecular weight of approximately 64,000. PMID- 2973352 TI - ATP-driven Ca2+ pump in the basolateral membrane of rat kidney cortex catalyzes an electroneutral Ca2+/H+ antiport. AB - An ATP-driven Ca2+ pump in the basolateral membrane of rat kidney cortex pumps Ca2+ out of the cell at the expense of MgATP (Km = 0.191 mM). This pump has a high affinity for free Ca2+ (26 nM). Vanadate, lanthanum, N-ethylmaleimide and calmodulin inhibitor R24571 inhibited this pump activity. Dimethyl[2 14C]oxazolidine-2,4-dione [( 14C]DMO) was entrapped in the vesicles in association with the ATP-driven Ca2+ influx. The ATP-driven Ca2+ influx was stimulated by the intravesicular acid pH and an upper convex Lineweaver-Burk reciprocal plot suggested two possible kinetics; one is that this Ca2+ pump is an allosteric enzyme with more than 1.72 H+ binding sites and another is the presence of two Ca2+ pumps with different affinities for H+. Valinomycin study indicated that the ATP-dependent Ca2+ transport by the BLMV was electroneutral and voltage independent. These results strongly suggest that the ATP-driven Ca2+ pump in the renal basolateral membrane catalyzes an electroneutral Ca2+/H+ antiport. PMID- 2973353 TI - DNA sequence analysis of the Olir2-76 and Ossr1-92 alleles of the Oli-2 region of the yeast Saccharomyces cerevisiae. Analysis of related amino-acid substitutions and protein-antibiotic interaction. AB - Petite deletion mapping helped to generate a fine-structure genetic map of the Oli-2 region of the mitochondrial genome of Saccharomyces cerevisiae. Here we report the DNA sequence analysis of the Oli-2 region from two drug-resistant alleles (Olir2-76 and Ossr1-92) which are located in the gene for subunit-6 of mitochondrial ATPase, in agreement with their genetic locations on the mitochondrial genome. An analysis of the corresponding amino-acid substitutions is also presented in the context of protein-antibiotic interactions. PMID- 2973354 TI - Reactivity of the thiol groups of Escherichia coli phosphofructo-1-kinase. AB - Modification of Escherichia coli phosphofructo-1-kinase (6-phosphofructokinase; EC 2.7.1.11) with several thiol modifying reagents led to a pseudo-first-order loss of activity that was associated with the modification of a single cysteine residue, identified as the cysteine at position 119 in the protein sequence. This cysteine was protected from reaction with vinyl pyridine, bromopyruvate, and dithionitrobenzoic acid by the substrate, fructose-6-P. In the crystal structure of the highly homologous phosphofructokinase from Bacillus stearothermophilus, cysteine 119 is sufficiently distant from the catalytic site to exclude a direct steric inhibition of the binding of substrate as a mechanism of inactivation for the modification. Thus, the inhibition is unlikely to be a direct one but to be the result of interference with the conformational change that is associated with fructose-6-P binding. A second thiol, position 283, was shown to be protected from reaction when the enzyme was in its native conformation. In contrast to the previously published sequence for the E. coli enzyme six cysteines as opposed to seven have been found both in enzyme from strain LE392 and in enzyme produced by a plasmid that was derived from pLC 16-4. The position in question, 75, was identified as phenylalanine. PMID- 2973355 TI - Tryptophan phosphorescence of the Ca2+-ATPase of sarcoplasmic reticulum. AB - Phosphorescence of protein tryptophan was analyzed in sarcoplasmic reticulum vesicles, and in the purified Ca2+ transport ATPase in deoxygenated aqueous solutions at room temperature. Upon excitation with light of 295 nm wavelength, the emission maxima of fluorescence and phosphorescence were at 330 nm and at 445 nm, respectively. The phosphorescence decay was multiexponential; the lifetime of the long-lived component of phosphorescence was approximately equal to 22 ms. ATP and vandate significantly reduced the phosphorescence in the presence of either Ca2+ or EGTA; ADP was less effective, while AMP was without effect. The quenching by ATP showed saturation consistent with the idea that the ATP-enzyme complex had a lower phosphorescence yield. Upon exhaustion of ATP, the phosphorescence returned to starting level. Significant quenching of phosphorescence with a decrease in phosphorescence lifetime was also caused by NaNO2, methylvinyl ketone and trichloroacetate, without effect on ATPase activity; this quenching did not show saturation and was therefore probably collisional in nature. PMID- 2973356 TI - Modulation of phosphofructokinase action by macromolecular interactions. Quantitative analysis of the phosphofructokinase-aldolase-calmodulin system. AB - The simultaneous effect of calmodulin and aldolase (D-fructose-1,6-bisphosphate D glyceraldehyde-3-phosphate-lyase, EC 4.1.2.13) on the concentration-dependent behaviour of muscle phosphofructokinase (ATP: D-fructose-6-phosphate 1 phosphotransferase, EC 2.7.1.11) has been analysed by means of a covalently attached fluorescent probe, gel penetration experiments, and using a kinetic approach. We found that calmodulin-induced inactivation of phosphofructokinase is suspended by addition of an equimolar amount of aldolase. This effect was attributed to an apparent competition of calmodulin and aldolase for the dimeric forms of kinase. Moreover, the direct binding of aldolase to calmodulin has also been demonstrated, which resulted in a significant decrease in the kcat value of the enzyme. The quantitative analysis of these interactions in the system phosphofructokinase-calmodulin-aldolase is presented. A possible molecular model for the modulation of phosphofructokinase action by macromolecular interactions is envisaged. PMID- 2973357 TI - Calcium binding protein changes of sarcoplasmic reticulum from rat denervated skeletal muscle. AB - Two Ca2+ sequestering proteins were studied in fast-twitch (EDL) and slow-twitch (soleus) muscle sarcoplasmic reticulum (SR) as a function of denervation time. Ca2+-ATPase activity measured in SR fractions of normal soleus represented 5% of that measure in SR fractions of normal EDL. Denervation caused a severe decrease in activity only in fast-twitch muscle. Ca2+-ATPase and calsequestrin contents were affected differently by denervation. In EDL SR, Ca2+-ATPase content decreased progressively, whereas in soleus SR, no variation was observed. Calsequestrin showed a slight increase in both muscles as a function of denervation time correlated with increased 45Ca-binding. These results indicate first that Ca2+-ATPase activity in EDL was under neural control, and that because of low Ca2+-ATPase activity and content in slow-twitch muscle no variation could be detected, and secondly that greater calsequestrin content might represent a relative increasing of heavy vesicles or decreasing of light vesicles as a function of denervation time in the whole SR fraction isolated in both types of muscles. PMID- 2973358 TI - [Conditions for antitumor cytotoxic T-lymphocyte formation and inhibition of their activity by T-suppressors in mixed culture of human lymphocytes and tumor cells]. AB - Conditions for antitumour autolytic T lymphocytes (CTL) induction during human mixed lymphocyte tumour cell culturing (MLTC), as well as the patterns of CTL activation abolition in the experimental system by suppressor cells were investigated. The responders used in MLTC were peripheral blood lymphocytes of colorectal cancer patients fractionated by means of multilayer Percoll gradients centrifugation (the density of layers being 1.077, 1.067, 1.056 g/ml). The cells of the second fraction collected in the density interphase of 1.077-1.067 g/ml (more than 90% of population belonging to T lymphocytes), when used as responders in MLTC, developed an autolytic activity against autologous and allogeneic tumour target cells. The cell of the first fraction (collected in the interphase of 1.067-1.056 g/ml) added to the second fraction cells at the beginning of MLTC, prevented the following CTL induction. The first fraction contained T-suppressor cells capable of strongly interfering with antitumour CTL activity. PMID- 2973359 TI - General practitioners' responsibilities. PMID- 2973360 TI - Effects of treating marrow with a CD3-specific immunotoxin for prevention of acute graft-versus-host disease. AB - Data from human clinical trials and animal experiments have suggested that T lymphocytes in donor marrow help to facilitate engraftment after allogeneic bone marrow transplantation, possibly through a suppressive effect on the immunity of the recipient. In previous studies marrows from HLA-identical donors were treated ex vivo with a mixture of eight monoclonal antibodies together with rabbit complement to achieve a 3-log depletion of T cells and CD3-negative lymphoid cells. Transplantation of this marrow was associated with a 27% actuarial risk of graft failure in leukemic recipients conditioned with cyclophosphamide (120 mg/kg) and 15.75 Gy fractionated total body irradiation. In the present study, we employed an anti-CD3 ricin A-chain-containing immunotoxin (64.1-A) together with 20 mM NH4Cl to achieve a selective 3-log depletion of CD3-positive cells. The patient entry criteria and pretransplant conditioning regimen were identical to those used in previous studies. Despite the differences in marrow treatment, the clinical outcome of the present study was similar to that obtained previously. Graft-versus-host disease (GVHD) was largely prevented without the need for post transplant immunosuppression, but two of the eight patients developed graft failure. These results indicate that CD3-negative cells have little or no ability to initiate GVHD. To the extent that graft failure in this study was not caused by stem cell damage or loss of CD3-negative cells during ex vivo processing of the marrow, it appears that the lymphoid cells required for facilitating allogeneic engraftment under these conditions are CD3-positive. PMID- 2973361 TI - Allogeneic bone marrow transplantation in multiple myeloma: a report of four cases. AB - Allogeneic bone marrow transplantation offers a new and promising form for treatment of multiple myeloma incurable with chemotherapy. We present four cases of advanced multiple myeloma given bone marrow transplantation from HLA-identical and MLC-negative sibling donors. One patient had a recurrent plasmacytoma 8 months later and one died 12 days after the transplantation whereas the other two are in good clinical remission 15 and 19 months post-transplantation. PMID- 2973362 TI - Technetium-99m HMPAO imaging in patients with basal ganglia disease. AB - Technetium 99m hexamethylpropylene-amine oxime (HMPAO) is trapped by cerebral grey matter and the basal ganglia on its first pass through the brain. To assess its potential for studying patients with diseases of the basal ganglia, a study of 15 normal volunteers and 32 patients with known or suspected basal ganglia disease have been investigated. Sixteen patients with idiopathic Parkinson's disease showed no abnormality of the basal ganglia and varying degrees of cerebral underperfusion consistent with their intellectual status. Eight patients with Huntington's chorea showed a characteristic pattern of reduced or absent caudate nucleus uptake. Patients with diseases affecting the basal ganglia, such as Fahr's disease, Wilson's disease and hemibalismus had varying degrees of basal ganglia underperfusion as demonstrated by an HMPAO scan. We believe that this new radiopharmaceutical for the demonstration of cerebral blood flow shows significant potential for the diagnosis and management of patients with basal ganglia disease. PMID- 2973363 TI - Antagonism between radiosensitizing agents. PMID- 2973364 TI - Oestrogen pre-treatment abolishes luteinising hormone-releasing hormone testosterone stimulation. AB - In patients with histologically confirmed prostate cancer, oestrogen priming with diethylstilboestrol (DES) (3 mg/day) for 4 weeks prior to the first injection of the LHRH agonist Zoladex (3.6 mg depot form) prevented any rise in the serum testosterone concentration. In contrast, in the groups pre-treated with DES, the first, but not subsequent, injections with Zoladex were associated with a marked surge in luteinising hormone. Treatment with DES beyond the time of the first administration of Zoladex did not provide further endocrinological advantage. Oestrogen priming for 1 month prior to treatment with Zoladex may prevent an exacerbation of signs and symptoms of prostate cancer. PMID- 2973365 TI - Suprapubic lithotripsy. PMID- 2973366 TI - The growth-associated neuronal phosphoprotein B-50: improved purification, partial primary structure, and characterization and localization of proteolysis products. AB - A reversed phase HPLC procedure is reported that has allowed the separation of the growth-associated, kinase C substrate protein B-50 [previously purified by isoelectric focussing (IEF)] into three components (1-, m- and rB-50). The minor form 1B-50 (probably a proteolysis product) gave a 24-residue N-terminal amino acid sequence, but the major and possibly native form (mB-50) (and also rB-50 which is probably formed during IEF) appeared to be N-terminally blocked. HPLC also separated B-60, the major proteolysis product of B-50, into three components, and the N-terminal sequence of the major B-60 was determined. HPLC peptide mapping of SAP digests of the various B-50 and B-60 protein confirmed their close relationship, and four SAP generated fragments also afforded sequence data. The amino acid sequences obtained (1B-50, B-60 and fragments) are all found in the recently predicted (based on nucleotide sequencing) B-50/GAP43 sequence (226 amino acids), further confirming the identity of B-50 and GAP43, and helping clarify the relationship of B-60 (starting at position 41 of the predicted sequence) to B-50. Correlation of amino acid analyses, SAP fragment data, and the predicted sequence provided additional information on the length of the translated products, including evidence that the N-terminus of the major (blocked) form of B-50 starts at position 1 (Met) of the predicted sequence. PMID- 2973367 TI - [Lipoprotein lipase (LPL): recall of the structure of LPL and first results obtained in the isolation of clones from human and rat genomic banks]. PMID- 2973368 TI - Utility of combining antigranulocyte with antileukocyte antibodies in differentiating Hodgkin's disease from non-Hodgkin's lymphoma. AB - The authors have reviewed their experience using the antigranulocyte marker, anti Leu-M1, in combination with the antileukocyte marker, PD7/26, applied to paraffin sections of malignant lymphomas difficult to subclassify using morphologic criteria. The study group consisted of 73 lymphomas; 53 cases of Hodgkin's disease and 20 cases of non-Hodgkin's lymphoma. Leu-M1 was expressed by the Reed Sternberg and Hodgkin's cells in 33 (62%) of the cases of Hodgkin's disease. The Reed-Sternberg and lymphocytic and histiocytic (L&H) cells in four cases of lymphocyte-predominant Hodgkin's disease were Leu-M1 negative. The Reed-Sternberg cells and L&H cells expressed leukocyte common antigen, utilizing the monoclonal antibody PD7/26, in seven (13%) of the 53 cases including the four cases of lymphocyte-predominant subtype (three nodular, one diffuse). The Reed-Sternberg like cells in four (20%) of the cases of non-Hodgkin's lymphoma were stained by anti-Leu-M1 whereas, in 12 cases (60%) these cells were stained by PD7/26. The combination of anti-Leu-M1 and PD7/26 provided more useful information than that provided by anti-Leu-M1 alone by providing immunologic support for the diagnosis of lymphocyte-predominant Hodgkin's disease and by identifying cases which stained with neither antibody. The authors interpret the immunologic findings in the latter cases as equivocal. These studies were most helpful in cases with many atypical cells and provided unequivocal support for the diagnosis of Hodgkin's disease in six of 11 cases of the "syncytial variant," a form of the nodular sclerosing type characterized by cohesive aggregates of Reed-Sternberg cells and lacunar variants, not uncommonly misdiagnosed using only morphologic criteria. PMID- 2973369 TI - Two partial remissions induced by an LHRH analogue in two postmenopausal women with metastatic breast cancer. AB - We report two cases of metastatic breast cancer occurring in two postmenopausal women. Treatment with a luteinizing hormone-releasing hormone (LHRH) analogue, D TRP6-LHRH, induced partial remission. The blood levels of the follicle stimulating hormone (FSH) and luteinizing hormone (LH) fell sharply under treatment, but the levels of estradiol, estriol, and estrone did not change significantly. We also described the putative mechanisms of action of the LHRH analog. PMID- 2973370 TI - Characterization of trimetrexate transport in human lymphoblastoid cells and development of impaired influx as a mechanism of resistance to lipophilic antifolates. AB - The purpose of this study was to characterize the transport properties of trimetrexate in WI-L2 human lymphoblastoid cells and determine the mode of resistance that had developed in a subline, WI-L2/TMQ, that was grown in increasing concentrations of trimetrexate. WI-L2/TMQ cells were 62-fold resistant to trimetrexate and 68- and 96-fold cross-resistant to the other lipophilic antifolates metoprine and piritrexim (BW 301U). No cross-resistance was observed with vincristine or doxorubicin, and sensitivity was not increased with 5 micrograms/ml of verapamil, indicating that it was not a typical multidrug resistance phenotype. WI-L2/TMQ exhibited a 2-fold increase in dihydrofolate reductase; however, this did not contribute significantly to the observed resistance, since these cells retained full sensitivity to methotrexate. Nor were there any kinetic alterations in dihydrofolate reductase toward trimetrexate or differences in the levels of thymidylate synthase. The major difference between the sensitive and resistant cell line was a 50% decrease in the influx rate of WI L2/TMQ cells which produced a corresponding decrease in cellular trimetrexate at the steady state. No difference in efflux rates was detected nor were there any differences in intracellular water or metabolism of trimetrexate. Additional characterization of trimetrexate transport in WI-L2 showed that influx was nonsaturable up to 5 mM extracellular trimetrexate, relatively insensitive to sodium azide, and exhibited a Q10 of 2.7. Influx was, however, inhibited in a dose-dependent manner by concentrations of p-chloromercuribenzylsulfonate above 10 microM. Efflux studies revealed a large nonexchangeable fraction of trimetrexate that was well above the dihydrofolate reductase binding capacity and varied depending on the initial level of cell-associated drug. The intracellular exchangeable trimetrexate concentration at the steady state was always several fold higher than the extracellular concentration. Retention of trimetrexate appeared to be coupled to some component of energy metabolism, since the presence of sodium azide stimulated this process by 2- to 3-fold. The data suggest that trimetrexate enters cells by passive diffusion but then is distributed and concentrated within the cell through more complex mechanisms which may involve energy coupling, compartmentation, or binding to macromolecules or organelles, although some type of carrier-mediated process cannot be ruled out.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973371 TI - Inhibition of rat ovarian [3H]thymidine uptake by luteinizing hormone-releasing hormone agonists: a possible mechanism for preventing damage by cytotoxic agents. AB - To investigate the mechanism involved in luteinizing hormone-releasing hormone (LHRH) agonists' protective effects against chemotherapy-induced ovarian damage, female rats were either implanted with 1-mg pellets of LHRH agonist Zoladex (LHRHz) or sham operated. All rats were implanted with osmotic minipumps loaded with [3H]thymidine 48 h before sacrifice in diestrus. Ovaries were combusted in a biological material oxidizer. Tritiated water was recovered in a special cocktail, and ovarian tritiated thymidine uptake (3HTU) was calculated. In five experiments, LHRHz significantly reduced ovarian 3HTU. This was observed 5 days after implanting LHRHz pellets. Ovarian 3HTU correlated significantly with serum estradiol, LH, and ovarian and uterine weights. Autoradiography showed that almost all ovarian 3HTU is by granulosa cells. These data suggest that LHRHz suppresses ovarian mitotic activity. Since cytotoxic agents preferentially destroy rapidly dividing cells, our findings may represent a mechanism for ovarian protection. PMID- 2973372 TI - Diastolic characteristics and cardiac energetics of isolated hearts exposed to volume and pressure overload. AB - The relation between diastolic dimensions and systolic function was studied in isolated rat hearts exposed to pressure (primary hypertension) and to volume load (hyperthyroidism). The hearts were arrested in diastole and cardiac dimensions analysed. Cardiac function was determined using an anterograde working heart preparation. Both models of cardiac overload produced significant left ventricular hypertrophy, however with different design. One concentric and one eccentric form of left ventricular hypertrophy were found in the spontaneously hypertensive rat obtained from two different breeders. An even more pronounced form of eccentric left ventricular hypertrophy was seen in hyperthyroid rats. The hearts with an eccentric form of hypertrophy had a higher maximal stroke volume when perfused at a high aortic pressure than those with a concentric form. At a low aortic pressure and hence a limited coronary perfusion cardiac performance was diminished in both types of spontaneously hypertensive rats but remained normal in hyperthyroid rats. Furthermore, oxygen consumption was reduced in all hypertrophied hearts compared with non-hypertrophied hearts. These data therefore suggest (a) that hearts having an eccentric type of hypertrophy may be in a more favourable situation, delivering a higher stroke volume for a given degree of myocardial fibre shortening; and (b) that the development of coronary vascular structural changes in spontaneously hypertensive rats, thereby increasing vascular resistance, leads to a decrease in left ventricular function at a low coronary perfusion pressure, whereas no such reduction was observed in hyperthyroid hearts, probably owing to a normal vascular resistance. PMID- 2973373 TI - Effect of flow partition on wall shear in a cast of a human coronary artery. AB - The effect of flow partition on wall shear was determined in a cast of a human coronary artery. A laser Doppler anemometer was used to measure the time varying velocities at selected sites close to the walls of the cast. Shear rates were derived by dividing the measured velocities at each site by the perpendicular distance from measurement site to the wall. At the bifurcation of the left main coronary artery into its anterior descending and circumflex branches the shear at sites near the outer walls of the two branches was more strongly affected by the flow partition than at sites on the flow divider and near the walls of the left main coronary artery. PMID- 2973374 TI - Ca, Mg-ATPase activity of permeabilised rat heart cells and its functional coupling to oxidative phosphorylation of the cells. AB - Isolated rat heart cells permeabilised by digitonin were examined as an experimental model to study heart bioenergetics. The cells showed good indices of oxidative phosphorylation (acceptor control ratio about 8 with pyruvate plus malate). The adenosine triphosphatase activity detected in the cells was high and was calcium dependent (optimum [free calcium] about 400 nmol.litre-1); magnesium was necessary for its full activity. Double reciprocal plot l/v vs 1/[free calcium] at physiological free calcium concentrations was linear, thus showing free calcium to be a substrate for the adenosine triphosphatase (Km for calcium about 149 nmol.litre-1). Double reciprocal plot 1/v vs 1/[ATP] was also linear, thus showing that the adenosine triphosphatase activity could be ascribed to a single enzyme. Oxidative phosphorylation and the ATPase activity of the cells appeared to be functionally coupled. This was manifested by apparent preference by oxidative phosphorylation for adenosine diphosphate supplied by the adenosine triphosphatase activity (Km 45 mumol.litre-1) to external adenosine diphosphate (Km 152 mumol.litre-1; p less than 0.02). Apparent preference by the adenosine triphosphatase activity for adenosine triphosphate supplied by mitochondria (Km 74 mumol.litre-1) to external adenosine triphosphate (Km 169 mumol.litre-1) was also manifested by a significant difference in Km values (p less than 0.05). PMID- 2973375 TI - Inhibitors of ADP-ribosyl transferase suppress the mitogenic actions exerted by tumour promoters, but not those evoked by peptide mitogens, in primary neonatal rat hepatocytes. AB - A significant stimulation of the 24-h (between day 4 and 5 in vitro) new DNA synthetic activity was elicited in primary neonatal rat hepatocytes kept in low calcium (0.01 mmol/l) HiWoBa2000 synthetic medium by the addition of a single dose (10(-10) mol/l) of each of several tumour promoters [i.e. 12-O tetradecanoylphorbol-13-acetate (TPA), phenobarbital, nafenopin, saccharin, teleocidin, benzoyl peroxide butylhydroxytoluene (BHT), dichlorodiphenyltrichloroethane (DDT), lindane, clofibrate and melittin]. Even hormones [e.g. epidermal growth factor (EGF), glucagon and insulin at 10(-10) mol/l] and EGF-like acting drugs (i.e. imidazole and indomethacin, at 10(-11) mol/l) similarly enhanced with respect to untreated controls the 24-h flow into S phase of the primary hepatocytes on condition, however, that the cells were incubated in a high- (i.e. 1.8 mmol/l) and not a low-calcium HiWoBa2000 medium. Xenobiotics, peptide mitogens and EGF-like acting drugs also enhanced the in vitro hepatocellular mitotic activity. The growth-stimulatory effects of the aforementioned eleven tumour promoters were entirely suppressed by the simultaneous addition to the growth medium of a fully effective dose (10(-4) - 10(-3) mol/l) of agents, such as 3-aminobenzamide (3-ABA), 3-methoxybenzamide (3 MBA) or nicotinamide (NA), that are known to inhibit the activity of ADP-ribosyl transferase (ADPRT). However, under the same conditions these inhibitors hampered neither the basal DNA synthetic and mitotic activities of spontaneously cycling hepatocytes nor the stimulation of the hepatocellular growth processes evoked by peptide mitogens and EGF-like acting drugs. Quantitative autoradiographic investigations showed that the incorporation of the ADP-ribose precursor and ADPRT substrate [3H]NAD into nuclear macromolecules of gently digitonin permeabilized hepatocytes was negligible in the untreated cultures, whereas it was strikingly and nearly steadily increased by a 2-, 8- and 24-h exposure to a fully mitogenic dose (10(-10) mol/l) of TPA, thereby revealing that an early, significant and roughly steady activation of the nuclear ADPRT had taken place in the phorbol ester-treated liver parenchymal cells. The simultaneous addition of 3 ABA (10(-4) mol/l) not only fully checked the mitogenic effects of TPA, but even suppressed about two-thirds of the TPA-elicited nuclear incorporation of [3H]NAD by the permeabilized hepatocytes, thus showing that a significant curtailment of the TPA-activated ADPRT did occur is association with the abatement of the mitogenic effects of TPA by this inhibitor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2973376 TI - Coronary artery vasoconstriction routinely occurs after percutaneous transluminal coronary angioplasty. A quantitative arteriographic analysis. AB - To determine whether percutaneous transluminal coronary angioplasty (PTCA) increases coronary artery luminal dimensions by stretching and injuring ("paralyzing") the smooth muscle of the arterial wall, we prospectively analyzed spontaneous changes and then intracoronary nitroglycerin-induced changes in segmental coronary artery diameters during the first 30 minutes after uncomplicated single-vessel PTCA in 10 patients. Five additional patients received intravenous nitroglycerin throughout the procedure to determine whether nitroglycerin could prevent vasoconstriction after PTCA. All of the patients were maintained on oral doses of diltiazem and aspirin at the time of the study. Coronary arteriography was performed at 2, 5, 15, and 30 minutes after PTCA and then 3 minutes after 300 micrograms i.c. nitroglycerin. Quantitative measurements (computerized edge-detection) were performed at each time, in coronary segments centered in the dilated segment, distal to the dilated segment, and in a control vessel not manipulated with the balloon catheter or guidewire. Progressive vasoconstriction (defined as a loss of diameter that was reversed by intracoronary nitroglycerin) was observed after PTCA in the dilated and distal segments (10 of 10 patients) but not in the control segment. The vasoconstriction in the dilated segment at 30 minutes (mean, 30 +/- 4%) was highly statistically significant compared with vasoconstriction at 2 and 5 minutes after PTCA (p less than 0.001) and compared with the control segment at 30 minutes (p less than 0.005). There was no significant loss of diameter after PTCA in the dilated segment in the five patients who received intravenous nitroglycerin. In conclusion, 1) spontaneous coronary artery vasoconstriction after PTCA occurs routinely at and distal to the site of balloon dilatation despite pretreatment with aspirin and calcium channel blockers; 2) coronary artery vasoconstriction after PTCA is rapidly reversed by intracoronary nitroglycerin and can be prevented by the continuous administration of intravenous nitroglycerin during and after the procedure; 3) these results are incompatible with the hypothesis that PTCA improves coronary luminal dimensions by arterial "paralysis"; and 4) these findings have implications concerning the etiology and prophylaxis of abrupt vessel closure after PTCA. PMID- 2973377 TI - Percutaneous transluminal coronary angioplasty improves survival in acute myocardial infarction complicated by cardiogenic shock. AB - Modest survival benefits have been reported in patients with acute myocardial infarction complicated by cardiogenic shock who were treated with early surgical revascularization or thrombolytic therapy. To determine whether coronary angioplasty improves survival, 87 patients with cardiogenic shock complicating acute myocardial infarction at the University of Michigan, Ann Arbor, Michigan, from 1975 to 1985 were retrospectively analyzed. Patients in group 1 (n = 59) were treated with conventional therapy; patients in group 2 (n = 24) were treated with conventional therapy and angioplasty. Extent of coronary artery disease, infarct location, and incidence of multivessel disease were similar between groups. Hemodynamic variables including cardiac index, mean arterial pressure, and pulmonary capillary wedge pressure were also similar. The 30-day survival was significantly improved for group 2 patients (50% vs. 17%, p = 0.006). Survival in group 2 patients with successful angioplasty was 77% (10 of 13 patients) versus 18% (two of 11 patients) in patients with unsuccessful angioplasty, (p = 0.006). The findings suggest that angioplasty improves survival in cardiogenic shock compared with conventional therapy with survival contingent upon successful reperfusion of the infarct-related artery. PMID- 2973378 TI - Cardiogenic shock complicating acute myocardial infarction. PMID- 2973379 TI - A genetic-diagnostic survey in an institutionalized population of 158 mentally retarded patients. The Viaene experience. AB - In this report we summarize the results of a genetic-diagnostic survey of an institutionalized population of 158 severely mentally retarded patients. The etiological study was based on a clinical genetic approach with special attention to dysmorphology and neurological findings. In 72 patients a constitutional cause of their mental impairment was found: a chromosomal abnormality in 21, a Mendelian disorder in 36 (autosomal recessive disorder: 23; autosomal dominant: 12; and X-linked recessive: 1), a MCA/MR syndrome in 9, and a CNS malformation in 6 patients. In 33 patients, a pre- or perinatal cause was found, and 20 patients presented a pre- or perinatal infection of the CNS. Finally, no etiological diagnosis was detected in 28 patients; 6 of them presented a hitherto unclassifiable type of familial mental retardation. PMID- 2973380 TI - Lymphocyte immunophenotyping of B-cell lymphomas: a flow cytometric analysis of neoplastic and nonneoplastic cells in 271 cases. AB - We have reviewed our experience with 271 B-cell lymphomas to determine the effectiveness of flow cytometry in the characterization of these malignancies. Flow cytometric immunophenotyping of the total lymphocyte and/or large lymphocyte populations confirmed the morphologic diagnosis of B-cell lymphoma in 92% of cases, which included 79% monoclonal and 13% surface immunoglobulin (SIg) negative lymphomas. Light chain monoclonality was most frequent in low grade and follicular center cell (FCC) lymphomas, while SIg-negative cases were most common in high grade and non-FCC types. Low grade lymphomas of all histologic types had high median percentages of neoplastic cells and low T-cell percentages. Conversely, high grade lymphomas exhibited lower and less uniform median percentages of B-cells, with higher numbers of T-cells and lower CD4/CD8 ratios than low grade lymphomas. Several differences in B- and T-cells were observed between specific high grade histologic types. FCC lymphomas with a diffuse pattern had lower percentages of CD4+ cells and lower CD4/CD8 ratios than cases with a follicular pattern. Thus, immunophenotypic differences were observed between histologic types or groups with known differences in clinical course and prognosis. We conclude that flow cytometry provides reliable information on neoplastic and nonneoplastic cells in lymph nodes involved by B-cell lymphomas. PMID- 2973381 TI - A reassessment of Holt's data on: "The question of lumbar discography". AB - This paper is a detailed critique of Holt's study entitled "The Question of Lumbar Discography." His finding of a 37% false positive rate in asymptomatic volunteers has been used to discredit lumbar discography as a valid diagnostic study in the investigation of low back pain patients with nonprolapsing disc disease of the lumbar spine. The authors show why Holt's paper should no longer be used as scientific and authoritative evidence against the use of discography by today's standards. PMID- 2973382 TI - [Epilepsy and pregnancy]. PMID- 2973384 TI - [An approach to the evaluation of the efficacy of anti-pertussis vaccination]. PMID- 2973383 TI - [Action of methoxybutropate in chronic bronchitis. Comparison with guacetisal]. PMID- 2973385 TI - [Effects of chronic administration of several calcium antagonists on pituitary release of TSH]. PMID- 2973386 TI - [Phosphocreatine in ischemic cardiopathy]. PMID- 2973387 TI - [An experimental clinical study on the bactericidal and bacteriostatic action of a disinfectant soap]. PMID- 2973389 TI - [Drug-induced hemopathies: thrombocytopenia]. PMID- 2973388 TI - [Changes in carbohydrate and lipid metabolism in chronic renal insufficiency]. PMID- 2973390 TI - [Therapeutic approach to elderly patients with rheumatoid arthritis]. PMID- 2973392 TI - British Diabetic Association, Medical and Scientific Section, autumn meeting. 1-3 September 1988, Stirling. Abstracts. PMID- 2973391 TI - [News about AIDS]. PMID- 2973393 TI - Irritant contact dermatitis and chrome-passivated metal. AB - An outbreak of hand dermatitis is reported amongst employees working on a new assembly line of an electronics factory. 24 out of 41 employees had signs of irritant contact dermatitis. The fingertips and the dorsal parts of the hands were especially affected. Mechanical factors in combination with zinc-chromium compounds from galvanised steel seemed to be responsible for the dermatitis and the dry skin. The use of an emollient solved the problem. PMID- 2973394 TI - Dyshidrotic eczema as an occupational dermatitis in metal workers. AB - Metalworkers exposed to metalworking fluids often encounter dermatological problems. In an epidemiological study of 286 metalworkers, 14% had dermatitis. Of these, 21 (54%) presented with the clinical appearance of dyshidrotic eczema. 3 out of these 21 operators had positive patch test reactions, 1 to nickel and 2 to formaldehyde and formaldehyde releasers, while 1 other was diagnosed as atopic. According to the literature, dyshidrotic eczema is strongly related to an atopic constitution and to contact sensitization, especially to nickel. However, our findings suggest that dyshidrotic eczema in metalworkers is a predominantly irritant dermatitis, mainly induced by exposure to soluble oils and unrelated to atopy. PMID- 2973395 TI - Skin temperature and skin symptoms among workers in the fish processing industry. AB - 196 workers employed in the fish processing industry participated in a survey of skin disorders. 156 (80%) had experienced skin problems during their work with fish on some occasions. The symptoms were itching, redness and stinging. Although the fingers are in direct contact with fish meat and juice, skin symptoms only seldom occur here, but instead almost exclusively on the forearms (70%) and the backs of the hands (26%). The skin temperature of the fingers and palms of the hands ranged from 17 degrees C to 20 degrees C, while the temperature on the backs of the hands and forearms ranged from 25 degrees C to 30 degrees C. Skin temperatures less than 20 degrees C abolish itch and reduce vasodilation by half. We suggest that the low temperature on the fingers affords protection against the development of some irritant skin reactions and that differences in skin temperature may be an important reason for the location of skin symptoms. PMID- 2973396 TI - Short communications. Allergic contact dermatitis from cobalt in the rubber industry. PMID- 2973397 TI - Contact dermatitis from glutaraldehyde. PMID- 2973398 TI - Occupational airborne contact dermatitis due to epoxy resin. PMID- 2973399 TI - Erythema multiforme-like reaction to "rosewood". PMID- 2973400 TI - Contact dermatitis from madder. PMID- 2973401 TI - Allergic contact dermatitis from "Varn Ecol - OJ wash" in a litho printer. PMID- 2973402 TI - Occupational allergic contact dermatitis from metallic mercury. PMID- 2973403 TI - Imidazole cross-sensitivity to sulconazole. PMID- 2973404 TI - Balloon angioplasty in the treatment of pulmonary hypertension caused by pulmonary embolism. AB - We present a 30-year-old man with pulmonary hypertension after pulmonary embolism. Pulmonary angiography showed multiple stenoses in the pulmonary vascular tree. We treated four of these stenoses by balloon angioplasty in three sessions. Pulmonary artery pressure was reduced from 90/25 mm Hg (mean 46) to 78/13 mm Hg (mean 35) with concomitant increase of aortic pressure from 105/60 mm Hg (mean 75) to 134/68 mm Hg (mean 90). Pulmonary perfusion scintigraphy showed increase of perfusion in the treated segments. Two procedures were followed by transient segmental pulmonary edema, but no other complications were noted. We conclude that balloon angioplasty is a promising method of lowering pulmonary artery pressure and improving pulmonary perfusion in suitable cases of pulmonary hypertension secondary to pulmonary embolism. PMID- 2973406 TI - [Seeding of autogenous endothelial cells to the inner surface of small-caliber dacron vascular prostheses: an experimental study in dogs]. PMID- 2973405 TI - Exercise-induced coronary artery spasm after percutaneous transluminal angioplasty. PMID- 2973407 TI - [Mechanism of photoregulation of carotenogenesis in Neurospora crassa: the use of mutants]. PMID- 2973408 TI - [Intermediate receptors]. PMID- 2973409 TI - Pharmacologic aids to intubation and the rapid sequence induction. AB - Endotracheal intubation usually can be performed in the emergency setting without the use of pharmacologic adjuncts. However, local airway anesthesia lessens patient discomfort, and the use of sedation and muscle relaxants occasionally may be necessary. Rapid sequence induction of general anesthesia adds benefits as well as risks to airway management; used in the circumstance of a full stomach combined with open eye injury or closed head injury associated with raised intracranial pressure, it should be practiced only by physicians appropriately trained and skilled at the procedure. PMID- 2973410 TI - Abscisic acid and water-stress induce the expression of a novel rice gene. AB - We have identified a novel rice gene, called RAB 21, which is induced when plants are subject to water-stress. This gene encodes a basic, glycine-rich protein (mol. wt 16,529) which has a duplicated domain structure. Immunoblots probed with antibodies raised against beta-galactosidase/RAB 21 fusion protein detect RAB 21 protein only in cytosolic cell fractions. RAB 21 mRNA and protein accumulate in rice embryos, leaves, roots and callus-derived suspension cells upon treatment with NaCl (200 mM) and/or the plant hormone abscisic acid (10 microM ABA). The effects of NaCl and ABA are not cumulative, suggesting that these two inducers share a common response pathway. Induction of RAB 21 mRNA accumulation by ABA is rapid (less than 15 min in suspension cells) and does not require protein synthesis, indicating that preformed nuclear and/or cytosolic factors mediate the response to this hormone. We have characterized the RAB 21 gene by determining the complete nucleotide sequence of a nearly full-length cDNA and corresponding genomic copy, and by mapping the start site of its major transcript. The proximal promoter region contains various GC-rich repeats. PMID- 2973413 TI - Importance of antigen specificity for complement-mediated lysis by monoclonal antibodies. AB - Lysis of human lymphocytes by autologous complement had been studied using a range of monoclonal antibodies against different antigens. Antigen specificity (and not antibody isotype) was the most important factor which influenced cell lysis and this could not be accounted for merely by differences in surface density between antigens. Three antigens with comparable surface density were studied in detail: CAMPATH-1 (lytic), major histocompatibility complex class I (lytic) and leukocyte common antigen (poorly lytic). C1q binding was roughly proportional to antibody binding and dependent on antibody isotype. However, the lytic antibodies were much better able to bind and activate whole C1 than the poorly lytic ones. This result would not have been predicted from traditional concepts of complement activation but can be interpreted in the light of models for C1 activation which involve Fc-Fc interactions, Fc-C1r2s2 interactions and a critical C1q stem-arm angle for C1 binding and activation. PMID- 2973411 TI - p36, the major cytoplasmic substrate of src tyrosine protein kinase, binds to its p11 regulatory subunit via a short amino-terminal amphiphatic helix. AB - Protein I is a hetero-tetramer which contains two copies each of p11 and p36. p36 (calpactin I, lipocortin II) is a major substrate of retrovirally encoded tyrosine protein kinases, while p11 modulates several Ca2+-induced properties also displayed by p36 alone. Here we have characterized the p11 binding site on p36 by fluorescence spectroscopy using porcine p36 labelled at cysteine 8 with the fluorophore Prodan (6-proprionyl-2-dimethylamino-naphthalene). We have used peptides of differing length from the amino-terminal domain of p36 to restrict the major binding site to the first 12 residues. Noticeable binding is still observed with a peptide containing only the first nine residues. Interestingly the N-terminal acetyl group of p36 forms a functional part of the p11 binding site. CD studies indicate that the binding region can form an alpha-helix, which seems to have amphiphatic properties when projected on a helical wheel. This structural element is also known for a calmodulin binding protein. Thus the question is raised whether other p11/calmodulin-related proteins interact with their target proteins via a similar mechanism. We also discuss how p11 could modulate p36 associated properties. PMID- 2973414 TI - Comparison of the structure of the murine interleukin 2 (IL 2) receptor on cytotoxic and helper T cell lines by chemical cross-linking of 125I-labeled IL 2. AB - The structure of the murine interleukin 2 receptor (IL 2R), on a cytotoxic (CTLL) and a helper (HT2) cell line, has been studied by a combination of chemical cross linking with 125I-labeled IL 2 and immunoprecipitation with an anti-receptor monoclonal antibody (7D4). In CTLL cells both methods detected the major 57-kDa IL 2-binding protein and in addition the cross-linking studies revealed the presence of a 70-75-kDa protein associated with the high-affinity receptor. In the HT2 cell line, however, immunoprecipitation studies revealed three additional proteins of 18, 22 and 37 kDa to the expected 50-kDa receptor protein. Again cross-linking studies demonstrated the presence of a 70-75-kDa protein, which was not immunoprecipitable with the 7D4 antibody. The low molecular polypeptides in HT2 cell were associated with the low-affinity receptor and represented most likely breakdown products of the 50-kDa protein. Whereas the 18- and 22-kDa proteins were involved in ligand binding, the 37-kDa fragment carried the epitope recognized by the 7D4 antibody. Comparative studies with two IL 2R antibodies, PC61 and 7D4, revealed that only PC61 inhibited the formation of the IL 2 alpha/beta chain complex, although both antibodies reportedly prevent the biological response to IL 2. It is speculated that the 37-kDa fragment, which reacts with the 7D4 antibody, might be involved in IL 2 signal transduction. Finally there was no evidence for the existence of a high molecular weight component of the IL 2R, previously described as gamma chain. In summary, the two chain structure of the IL 2R has been confirmed for both murine cell lines with some heterogeneity of the alpha chain. The possibility was raised that a 37-kDa fragment of the alpha chain plays a role of in signal transduction. PMID- 2973416 TI - Expression of CD23 antigen is not necessary for human 12-kDa B cell growth factor mediated B cell proliferation. AB - In an attempt to understand the relationship between the CD23 antigen and the human 12-kDa B cell growth factor (BCGF) receptor, we have undertaken studies to define the biological efficacy of 12-kDa-BCGF on CD23+ (Raji) and CD23- (P3HR1) human Burkitt B cell lines. Our results show that recombinant 12-kDa-BCGF can induce efficient [3H]thymidine incorporation and proliferation in cells of either phenotype. These results strongly suggest that a functional receptor for the 12 kDa-BCGF may exist independently of the CD23 molecule. PMID- 2973415 TI - Organization of the human CD3 locus on chromosome 11. AB - Three CD3 genes, gamma, delta, epsilon, map to human chromosome 11 band q23. A 75 kb region of 11q23 containing all three genes has been characterized by field inversion gel electrophoresis and molecular cloning. The CD3 genes lie within a stretch of 50 kb of DNA, orientated 3'-CD3 gamma-5': 5'-CD3 delta-3': 3'-CD3 epsilon-5'. Only 1.6 kb separate the CD3 gamma and CD3 delta genes, while the CD3 delta and CD3 epsilon genes are approximately 22 kb apart. CD3 gene expression is one of the first events during the maturation of the thymocyte, and therefore the clustering of CD3 genes may define a chromatin domain which is activated early in T cell development. PMID- 2973417 TI - Calpactin-like proteins in human spermatozoa. AB - Polyclonal antibodies directed against human calpactin I (p36) and calpactin II (p35) have been employed to investigate the distribution of calpactin-like proteins in human spermatozoa. Calpactins are a family of Ca2+-regulated cytoskeletal proteins that are major substrates of oncogene and growth factor receptor protein tyrosine kinases. The existence of a Triton-soluble 37-kDa protein antigenically related to calpactin II from somatic cells was revealed by Western blot analysis of human sperm extracts. The 37-kDa protein was not released from spermatozoa after experimental induction of the acrosome reaction by A23187 and Ca2+. Treatment of sperm homogenates with an EGTA-containing buffer partially solubilized the 37-kDa protein from the corpuscolate matter. Indirect immunofluorescence microscopy showed that anticalpactin II binds specifically to the sperm tail and to a band-like structure encircling the sperm head at the equatorial segment. In contrast, antibodies to calpactin I were found to bind to the tail midpiece, but failed to bind to Western blots of sperm proteins. This is the first immunological and biochemical report on the presence of calpactin proteins in a germ cell, the human spermatozoon. PMID- 2973418 TI - [75th anniversary of the Department of Pharmacology with General Toxicology and a Clinical Pharmacology Course of the Leningrad Health and Hygiene Medical Institute]. PMID- 2973419 TI - An example of substrate channeling between co-immobilized enzymes. Coupled activity of myosin ATPase and creatine kinase bound to frog heart myofilaments. AB - In myofilaments obtained by Triton X-100 lysis of frog heart cells in high ionic strength medium, the activity of bound creatine kinase cannot be detected by a coupled enzymatic assay. ATP is channelized toward myosin ATPase, through the unstirred layer near myofilaments and cannot diffuse into the bulk solution. Model systems based upon the coupled kinetics of enzymes co-immobilized on the same surface may explain this behaviour. This may also account for why myofilament-bound creatine kinase is more efficient than free enzyme in the cytosol for the physiological recycling of ADP into ATP. PMID- 2973420 TI - CA 125 in peritoneal fluid and serum of patients with endometriosis. AB - Serum and peritoneal fluid collected from 48 patients at laparoscopy was assayed for CA 125. Geometric mean serum CA 125 was similar in 21 patients with predominantly early stage endometriosis (13.4 U/ml; confidence intervals (C.I.) 10.8-16.7) and 27 patients without endometriosis (12.9 U/ml; C.I. 9.7-17.2). Mean peritoneal fluid CA 125 was higher in the endometriosis patients (mean 44.4 U/ml; C.I. 34.5-57.2) than controls (35.7 U/ml; C.I. 30.0-42.2), but the difference was not significant. Peritoneal fluid volume was increased in endometriosis patients (p = 0.013), and only after correction for peritoneal fluid volume did total antigenic concentration in aspirated peritoneal fluid show significant elevation (p = 0.0015) in endometriosis patients (mean 340 U/ml; C.I. 212-547) compared with controls (125 U/ml; C.I. 85-184). These results suggest that although CA 125 may be expressed in peritoneal fluid in endometriosis, it is unlikely to be of use as a marker of disease activity in peritoneal fluid obtained by culdocentesis. PMID- 2973421 TI - Experimental and clinical studies on the anticancer agent, doxifluridine (5' DFUR), Satellite symposium to the 25th annual meeting of the Japan Society for Cancer Therapy. Sapporo, Japan, 7 October 1987. Abstracts. PMID- 2973422 TI - Antibodies to insulin-like growth factor I receptors in diabetes and other disorders. AB - A newly developed immunoprecipitation assay, with 125I-labeled highly purified human placental insulin-like growth factor I (IGF-I) receptor, was used to search for IGF-I-receptor antibodies in human sera. Eleven of 141 patient sera tested (7.8%) immunoprecipitated labeled IGF-I receptor. Immunoprecipitation was comparable with sera and IgG prepared from these sera. Seven of the 11 sera (3 of 31 with rheumatic disorders, 3 of 48 with non-insulin-dependent diabetes, and 1 of 52 with insulin-dependent diabetes) failed to inhibit IGF-I binding to human placental membranes and thus contained non-binding-inhibitory IGF-I-receptor antibodies. Their pathophysiological function remained uncertain. The remaining 4 sera (2 of 3 with type B severe insulin resistance, 1 of 7 with polycystic ovary syndrome (PCO), and 1 of 31 with rheumatic disorders) inhibited IGF-I binding. Plasma IGF-I concentrations were elevated (663 and 802 ng/ml, respectively) in 2 patients (1 with PCO and another with systemic lupus erythematosus) with binding inhibitory IGF-I-receptor antibodies, suggesting IGF-I resistance that was probably mediated by the IGF-I-receptor antibodies. In conclusion, we identified two species of human IGF-I-receptor antibodies. Sera from 7 of 141 patients tested contained IgG autoantibodies that bound to the IGF-I receptor at a locus different from the IGF-I binding site and did not inhibit IGF-I binding. Sera from 4 of 141 patients contained antibodies that bound to the IGF-I receptor at or near the IGF-I binding site, inhibited IGF-I binding, and probably caused IGF I resistance. PMID- 2973423 TI - A self-glucosylating protein is the primer for rabbit muscle glycogen biosynthesis. AB - In this paper we elucidate part of the mechanism of the early stages of the biosynthesis of glycogen. This macromolecule is constructed by covalent apposition of glucose units to a protein, glycogenin, which remains covalently attached to the mature glycogen molecule. We have now isolated, in a 3500-fold purification, a protein from rabbit muscle that has the same Mr as glycogenin, is immunologically similar, and proves to be a self-glucosylating protein (SGP). When incubated with UDP-[14C]glucose, an average of one molecular proportion of glucose is incorporated into the protein, which we conclude is the same as glycogenin isolated from native glycogen. The native SGP appears to exist as a high-molecular-weight species that contains many identical subunits. Because the glucose that is self-incorporated can be released almost completely from the acceptor by glycogenolytic enzymes, the indication is that it was added to a preformed chain or chains of 1,4-linked alpha-glucose residues. This implies that SGP already carries an existing maltosaccharide chain or chains to which the glucose is added, rather than glucose being added directly to protein. The putative role of SGP in glycogen synthesis is confirmed by the fact that glucosylated SGP acts as a primer for glycogen synthase and branching enzyme to form high-molecular-weight material. SGP itself is completely free from glycogen synthase. The quantity of SGP in muscle is calculated to be about one-half the amount of glycogenin bound in glycogen. PMID- 2973424 TI - [Functional anatomy of the left ventricle in hypertensive subjects. Correlations with the clinical severity of hypertension]. AB - Using computerized M-mode echocardiography we have investigated 58 hypertensive subjects in order to assess whether a correlation could be demonstrated between left ventricular changes induced by hypertension and age of the patients, the duration and severity of hypertension, and damage to other target organs. Various morphological changes of the left ventricle were detected: 14 patients (24%) had concentric hypertrophy of the left ventricle, 12 (20%) had asymmetric septal hypertrophy, 5 (8%) had dilated left ventricle without hypertrophy. Left ventricular mass was increased, when compared to normal controls in 24 patients (41%). With respect to functional abnormalities, the peak lengthening rate of left ventricular dimension in diastole was decreased (+dD/dt less than s-1) in 25 patients (43%). Eight of these patients (14%) also had depressed peak shortening rate of left ventricular diameter in systole (-dD/dt less than 1.9 s-1). Hypertensive retinopathy was present in 23 patients (39%) and impairment of renal function in 8 (14%). Left ventricular mass and systolic and diastolic parameters of left ventricular function did not correlate significantly either with the age of the patients, or with the duration and severity of hypertension, or with the damage present in target organs other than the heart. Left ventricular mass was inversely correlated with the index of left ventricular relaxation (r = -0.53; P less than 0.001), whereas neither the latter nor left ventricular mass were correlated with peak systolic stress. Instead, peak systolic stress was inversely correlated with peak shortening rate of left ventricular diameter, an index of systolic function (r = -0.50; P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973425 TI - Beta-endorphin in the brainstem, pituitary, and spinal fluid of infants at autopsy: relation to sudden infant death syndrome. AB - Immunohistochemical localization of beta-endorphin was studied in the pituitaries and medullas of forty human infants at autopsy. beta-Endorphin immunoreactivity was found in anterior pituitary cells in all cases. In the medulla, beta endorphin immunoreactivity was found in the neurons of the medial and lateral cuneate nuclei in ten out of the forty cases. In eight of these ten cases, the infants died of causes other than sudden infant death syndrome (SIDS). Only two of 25 SIDS cases had demonstrable beta-endorphin in the brainstem nuclei. Beta endorphin levels in the spinal fluids of all the cases showed no correlation to cause of death, age or gender. PMID- 2973426 TI - Small-scale purification of bacteriophage lambda DNA by an airfuge centrifugation step in cesium chloride gradients. AB - A rapid and efficient procedure for purifying bacteriophage lambda DNA is described. This small-scale purification involves isolation of bacteriophage particles on cesium chloride gradients. Using an Airfuge ultracentrifuge, the centrifugation step can be readily achieved in 90 minutes. The method allows a 1 day purification of up to 12 independent lambda DNA (20-40 micrograms each). The recovered DNA, essentially devoid of RNA and DNA contaminants, is efficiently cut by restriction endonucleases and can serve as starting material for the ligation of DNA fragments in other cloning vehicles. PMID- 2973427 TI - Non-small cell lung carcinomas with neuroendocrine features. A light microscopic, immunohistochemical and ultrastructural study of 11 cases. AB - Eleven resected primary lung carcinomas classified as large cell carcinomas or squamous cell carcinomas, but showing some microscopic resemblances to bronchial carcinoid and small cell carcinoma, were studied. All cases were neurone-specific enolase and protein gene product 9.5 positive, indicating neuroendocrine differentiation. Staining for bombesin, C-terminal peptide of human pro-bombesin and chromogranin was positive in some cases. Electron microscopy showed dense core granules in six of seven cases investigated, the remaining case showing small granules of uncertain nature. All but one patient died within 15 months after operation. These data indicate that neuroendocrine differentiation in non small cell carcinomas of the lung may in some cases be suspected on routine histology. The follow-up data suggest that the identification of these cases might have implications for prognosis and therapy, and consequently for diagnostic lung tumour classification. PMID- 2973428 TI - Antigenic phenotype of Langerhans cell histiocytosis: an immunohistochemical study demonstrating the value of LN-2, LN-3, and vimentin. AB - In order to determine the antigenic phenotype of the proliferating cells in Langerhans cell histiocytosis (LCH), we studied 15 such examples by using formalin- and B5-fixed, paraffin-embedded tissues. We used a panel of antibodies that are known to react with lymphocyte- and histiocyte-associated antigens. These included LN-1, LN-2, and LN-3 monoclonal antibodies (MoAbs), MoAbs to leukocyte common antigen (LCA), Leu-M1 antigen, vimentin, and epithelial membrane antigen (EMA), as well as polyclonal antibodies to lysozyme and S100 protein. The antigens encountered most frequently in LCH cells were S100 protein (93% of cases), vimentin (86%), and those detected by LN-2 (80%) and LN-3 (82%). Lysozyme was detected focally in two cases and diffusely in one case. The LCH cells were negative for LN-1, LCA, Leu-M1, and EMA. There was only one specimen in which S100 protein was not demonstrated; in this case, LN-3, vimentin, and T6 on frozen section were positive. The phenotype of LCH cells is similar to that of Langerhans' cells and interdigitating histiocytes. Our results demonstrate the value of using a panel of antibodies, including anti-vimentin MoAb, LN-2, and LN 3 for the immunophenotypic diagnosis of LCH in addition to an antibody to S100 protein. PMID- 2973429 TI - Monoclonal antibody Leu-22 (L60) permits the demonstration of some neoplastic T cells in routinely fixed and paraffin-embedded tissue sections. AB - Monoclonal antibody Leu-22 (L60) detects a T cell-associated antigen which is stably expressed in routinely fixed and paraffin-embedded tissue sections. We investigated the utility of monoclonal antibody Leu-22 to immunophenotype routinely processed lymphoid neoplasms by determining its reactivity in 105 archival pathologic specimens of lymphoid neoplasia that had been previously immunophenotyped by standard cell suspension and frozen tissue section techniques. Monoclonal antibody Leu-22 reacted with 69% of T cell non-Hodgkin's lymphomas (NHLs), including cases belonging to each of the major clinicopathologic categories, and with 22% of B cell NHLs, but did not react with the Reed-Sternberg (RS) cells of Hodgkin's disease (HD). We concluded that monoclonal antibody Leu-22 reacts preferentially but not exclusively with T cell NHLs. Therefore, we performed parallel analyses of the same 105 cases with monoclonal antibodies leukocyte common antigen (LCA), Leu-M1, LN1, and LN2, which detect various paraffin-resistant antigens, and of 80 of these cases with monoclonal antibody UCHL1, which detects a paraffin-resistant T cell-associated antigen. UCHL1 reacted with 61% of the T cell NHLs studied. Sixty-nine percent of T cell NHLs expressed the LCA+, Leu-22+ or Leu-M1+, LN1- phenotype and 47% of B cell NHLs expressed the LCA+, Leu-22-, Leu-M1-, LN1+ phenotype. These phenotypes had a false-positive rate of only 7%. The substitution of UCHL1 for Leu-22 or the combined use of UCHL1 and Leu-22 in this panel did not improve our ability to correctly predict the T cell phenotype of these lymphoid neoplasms. LN1 and LN2 reacted with 13% and 56% of T cell NHLs, respectively, and LN2 reacted with RS cells in 85% of cases of HD. In summary, our results demonstrate that the judicious use of monoclonal antibody Leu-22 in combination with other selected commercially available monoclonal antibodies permits the determination of the B cell or T cell origin of a high proportion of NHLs, and is helpful in the differential diagnosis between HD and NHL among cases that have been routinely fixed and paraffin-embedded. PMID- 2973430 TI - Trisomy 21q223 and Down's phenotype correlation evidenced by in situ hybridization. AB - Two cases of trisomy 21q223 with the Down's phenotype were analysed by in situ hybridization with specific probes previously located in the sub-bands 21q221 (SOD-A) and 21q223 (BCEI and COL6A). These studies give evidence that the clinical picture of Down's syndrome is at least to a great extent correlated with trisomy for the 21q223 band. PMID- 2973431 TI - Antigen-bound C3b and C4b enhance antigen-presenting cell function in activation of human T-cell clones. AB - The effect of complement fragments C3b and C4b, on the triggering of antigen specific human T-cell clones by Epstein-Barr virus-transformed human lymphoblastoid B cells (LCL) when these fragments are covalently coupled to the antigen tetanus toxin (TT) is described. TT was chemically cross-linked to purified C3b [(TT-C3b)n], C4b [(TT-C4b)n] or bovine serum albumin [(TT-BSA)n] as a control. T-cell activation was quantified by tritiated thymidine incorporation and 51Cr release. (TT-C3b)n and (TT-C4b)n induced proliferative responses comparable to (TT-BSA)n but at 18-25 and 4-6 lower concentrations, respectively. This enhancing effect required the covalent cross-linking of the complement fragments to the antigen and involved intracellular processing of the latter by LCL. Antigen presentation was similarly enhanced when measuring the cytotoxic activity of a helper T-cell clone against LCL previously pulsed with (TT-C3b)n or (TT-C4b)n compared with (TT-BSA)n. Binding studies, carried out on LCL using TT radiolabelled with 125I before cross-linking, indicated that (TT-C3b)n and (TT C4b)n gave three- to four-fold more binding than (TT-BSA)n. Addition of antibodies against CR1 and CR2 or proteolytic removal of these complement receptors with trypsin inhibited by about 60% the enhancing effect of TT-bound C3b and C4b in both binding and functional assays. These results indicate that binding of C3b or C4b to antigen enhances antigen-specific proliferative and cytotoxic responses of T cells by targeting opsonized antigen onto complement receptors CR1 and CR2 of LCL. The putative significance of these findings in terms of regulation of immune responses by complement is discussed. PMID- 2973432 TI - The autonomy of CD8+ T cells in vitro and in vivo. AB - Experiments have been carried out in vitro and in vivo to determine to what extent CD8+ T cells in the rat can function independently of any helper activity from CD4+ cells. We have identified the culture conditions required for the autonomous proliferation of CD8+ T cells in the rat mixed leucocyte culture (MLC) and in particular have studied both the kinetics of the response and the effect of replacing the homologous serum, used in our previous MLC experiments, with fetal calf serum (FCS). The results obtained using FCS show that, early in the MLC, CD8+ T-cells proliferate at a comparable rate to the CD4+ subset but that, within 48-72 hr, the proliferation rate of the CD8+ cells ceases to increase with time. In contrast, the proliferation of the CD4+ T cells appears to be limited only by the exhaustion of the culture medium. The results also show that the proliferative responses of both CD4+ and CD8+ T cells are inhibited in homologous serum but that it is the CD8+ subset that is more affected. When CD8+ T cells, in homologous serum, are co-cultured with irradiated CD4+ T cells the proliferative activity is increased, indicating that the helper activity of the CD4+ T cells can over-ride the inhibitory effect of the serum. In vivo we have compared the abilities of injected CD4+ and CD8+ T cells to mediate rejection of skin allografts on nude rats. Grafts were rejected more rapidly on recipients of low doses of CD4+ T cells than on rats given 200 times as many CD8+ T cells. Thoracic duct lymphocytes (TDL) obtained 5 weeks after CD8+ T-cell injection always contained a population of CD4+ T cells, even when the injected CD8+ T-cell inoculum contained less than 0.1% CD4+ cells as contaminants. Evidence was obtained that these CD4+ T cells found in TDL displayed alloreactivity in MLC. Further, the intentional injection of very low doses of CD4+ cells led, after 5 weeks, to frequencies of CD4+ T cells, in thoracic duct lymph, equal to that obtained by the injection of 200 times as many cells of the same phenotype. It appears that, in T-cell-deficient rats, CD4+ cells can expand over 2000 times in a few weeks. Such expansion may explain the relatively slow rejection of skin allografts observed in these experiments when nude rats were injected with putatively pure populations of CD8+ T cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2973434 TI - Alterations in helper-inducer and suppressor-inducer T-cell subsets in human neonatal blood. AB - The distribution of T-helper cell subsets of human umbilical cord lymphocytes was compared with that of adult peripheral blood lymphocytes. Cytofluorometric analysis revealed a similar Leu 3/Leu 2 (helper/suppressor) ratio in neonates compared with adults. Total T-cell numbers were slightly decreased in neonatal blood. The Leu 3-positive-4B4-positive cell (helper-inducer) subset, however, was markedly reduced in neonatal blood, while the Leu 3-positive-2H4-positive cell (suppressor-inducer) subset was increased compared with that of normal adults. These findings may contribute to the poor help and enhanced suppression observed in vitro with neonatal T cells. PMID- 2973433 TI - gp 58/68, a parasite component that contributes to the escape of the trypomastigote form of T. cruzi from damage by the human alternative complement pathway. AB - A glycoprotein of apparent molecular weight 58,000 (unreduced)/68,000 (in its reduced form) (gp 58/68), which is one of the fibronectin/collagen receptors of Trypanosoma cruzi, was purified to homogeneity from the trypomastigote forms of the Tehuantepec and Y strains of the parasite. Purified gp 58/68 inhibited formation of cell-bound and fluid-phase alternative pathway C3 convertase in a dose-dependent fashion, as assessed using purified human complement components. Gp 58/68 differed from the human regulatory proteins H, DAF, MCP and CR1 and from previously reported regulatory proteins on the parasite membrane in that it was unable to enhance decay-dissociation of preformed alternative pathway C3 convertase sites, did not serve as a co-factor for I-mediated cleavage of C3b and had no inhibitory activity on the classical pathway convertases. The inhibitory effect of gp 58/68 was most likely dependent on an interaction of the protein with factor B rather than with C3b. Gp 58/68 provides trypomastigotes with an additional potential mechanism for escaping complement lysis by the human alternative pathway. PMID- 2973435 TI - Suppressor Lyt 2+ T-cells demonstrated in mice late after thermal injury. AB - Biological properties of suppressor cells mediating second suppressive state (S SupS) of IFN-gamma responsiveness in mice following thermal injury were investigated by their ability to inhibit [3H]thymidine incorporation by allogeneic cell-stimulated mononuclear cells (MNC) prepared from spleens of unburned normal mice (N-mice). Kinetic studies indicated that the suppressor cells appeared at approximately 15 days post-burn and persisted until 40 days after thermal injury. This cell population from the spleens of thermally injured mice (TI-mice) was inactivated by anti-Thy 1.2 monoclonal antibody (mAb) plus complement (C). Treatment with antimouse immunoglobulin and anti-Lyt 1.2 mAb followed by C failed to abrogate the activity. However, it could be eliminated by treatment with anti-Lyt 2.2 mAb and C. In addition, when Thy 1+ or Lyt 2+ T cells were depleted from spleen cells containing suppressor cell activities by treatment with mAbs and C, the suppressor activity for IFN-gamma production by N mice splenic MNC stimulated with concanavalin A was also eliminated. However, no alteration in the suppression of IFN production was observed when this splenic cell population from TI-mice was treated with anti-Lyt 1.2 mAb and anti-mouse Ig followed by C. Since it has been reported [1] that the suppressor cell in spleens of TI-mice could neither be removed by plastic adherence nor by carbonyl-iron treatment, this secondarily-generated suppressor cell was concluded to be a T cell possessing the Lyt 1-2+ phenotype. The present report describes the demonstration, following a generation of suppressor macrophages [2], of suppressor Lyt 2+ T cells spleen of mice late after thermal injury. PMID- 2973437 TI - Betamethasone dipropionate (0.05)% plus chinoform (3%) cream against betamethasone valerate (0.1%) plus chinoform cream (3%) in secondarily infected dermatoses. PMID- 2973436 TI - Human retrovirus-related synthetic peptides inhibit T lymphocyte proliferation. AB - Immunosuppression frequently accompanies infections with the human retroviruses HTLV-1 and HIV. Previous studies have shown that UV-inactivated and detergent disrupted preparations of either virus can produce immune dysfunction in vitro although the active component of such preparations has not yet been identified. We now report that synthetic peptides corresponding to a conserved sequence within the transmembrane envelope proteins of both HTLV-1 (CS-1) and HIV (CS-3) are capable of suppressing the in vitro proliferative responses of human T lymphocytes to mitogenic or allogeneic stimulation. CS-1 inhibits proliferation in response to stimulation by anti-CD3 antibody by up to 57% a concentration of 3 nmol per culture well. CS-3 inhibits by up to 49% at a similar concentration. CS 1 and CS-3 inhibit two-way mixed leukocyte culture reactions by 81 and 88%, respectively, at the 3 nmol per well concentration. The inhibition by such human retrovirus-related peptides appears to be T cell-specific in that B cell proliferation in response to stimulation with anti-IgG is not affected by the CS 3 peptide. The release of such immunosuppressive peptides may thus play a role in the pathogenesis of human exogenous retroviral disease such as AIDS. PMID- 2973439 TI - Atrial natriuretic factor in specific brain areas of spontaneously hypertensive rats. AB - Atrial natriuretic peptides (atrial natriuretic factor, ANF) are present in a great number of brain areas inside and outside of the blood-brain barrier. The pattern of distribution implies the involvement of ANF in different physiological functions, such as blood pressure regulation, electrolyte and fluid homeostasis, and modulation of the neuroendocrine system. To further investigate a possible involvement of central ANF in spontaneous hypertension, we measured levels of ANF in 18 selected, microdissected brain areas of prehypertensive (4-week-old) and hypertensive (12-week-old) spontaneously hypertensive rats (SHR) and their normotensive control, Wistar-Kyoto rats (WKY), by radio-immunoassay. ANF was significantly decreased in seven brain areas in SHR at both ages investigated; the most pronounced decreases were found in the subfornical organ, in the perifornical and periventricular hypothalamic nuclei, and in the medial preoptic nucleus. In addition, in young SHR ANF was significantly decreased in the organum vasculosum laminae terminalis and increased in the median eminence. After the development of hypertension, a significant decrease of ANF could be detected in four more brain areas (bed nucleus of the stria terminalis, paraventricular and arcuate nuclei, dorsal raphe nucleus) of SHR, as compared with normotensive controls, and the increase in the median eminence was no longer detectable. These results suggest a role for ANF in genetic hypertension and the specific importance of certain brain regions. PMID- 2973440 TI - Immunoglobulin E and immunoglobulin G subclass distribution in vivo and relationship to in vitro generation of interferon-gamma and neopterin in patients with severe atopic dermatitis. AB - In vitro interferon-gamma (IFN gamma) and neopterin generation by peripheral blood mononuclear cells (PBMC) from 15 patients with severe atopic dermatitis (AD) and 10 healthy controls was investigated. A significant proportion of patients had an impaired capacity to secrete IFN gamma after phytohemagglutinin (PHA) stimulation in vitro and therefore IFN gamma production was significantly lower compared to controls. Neopterin generation in vitro did not differ significantly from that of controls and no correlation between in vitro IFN gamma and neopterin production could be observed in either group. Analysis of serum IgG subclass distribution showed that patients with AD. had increased IgG4 serum concentrations while IgG1, IgG2 and IgG3 levels did not differ significantly from those of controls. Surface marker analysis revealed increased numbers of CD23+ lymphocytes in patients with AD which was positively correlated with the serum IgG4 and IgE concentration. Furthermore, a significant correlation was found between IFN gamma generation in vitro and IgE and IgG4 concentration in vivo in AD. The data suggest that a possible dysregulation of IFN gamma, interleukin-4 or other lymphokine interleukin-4 or other lymphokine production may be related to increased IgE and IgG4 production and seems to be an important factor in the pathogenesis of AD. PMID- 2973441 TI - Behaviour of guinea pig T cells stimulated by antigen, allo-antigen and mitogen. AB - The expression of major histocompatibility (MHC) antigens on guinea pig T cells was used as a functional marker for lymphocyte activation. Antigen-stimulated lymphocytes were recovered from guinea pigs responding to the contact sensitizer DNFB, and isolated T cells were then phenotyped using a new antiguinea pig monoclonal antibody, MSgp7. The level of expression of MHC class II, as defined by the monclonal antibody, MSgp8, was increased on T cells recovered 4 days after sensitization, as compared with unsensitized controls. The value of this experiment was extended by measuring MHC class II expression on T cells stimulated in vitro by the mitogen concanavalin A, where a clear increase in MSgp8 binding was also observed. Confirmation of the specificity of MSgp8 for guinea pig MHC class II antigens was achieved by studying the inhibitory capacity of this antibody on an MHC class II restricted mixed leucocyte reaction. The combination of antibodies MSgp7 and MSgp8 with flow cytometry could be applied to other guinea pig experimental models to quantitate the expression of MHC class II antigens on T cells to determine their putative value in disease manifestation. PMID- 2973438 TI - Salt sensitivity in blacks. Salt intake and natriuretic substances. AB - Accumulating evidence suggests that hypertension in blacks is manifested in part by impaired renal excretion of salt. Consequently, this study was performed to determine if hypertensive and normotensive black subjects differ in their ability to generate known natriuretic substances. Fourteen normotensive and 11 hypertensive blacks were maintained on constant metabolic diets containing either 40 or 180 mmol of salt per day for 14 days each. During the last 4 days of each salt intake period, urine was collected for measurement of sodium, dopamine, and norepinephrine. On the last day of each 14-day dietary period, blood pressures were measured, blood was collected for measurement of plasma atrial natriuretic factor (ANF) and aldosterone, and urine was collected over 2 hours for measurement of prostaglandin E2 (PGE2). Both the normotensive and the hypertensive groups manifested salt sensitivity; their mean arterial pressure rose by 7 +/- 0.2 and 6 +/- 0.2%, respectively, when salt intake was increased from 40 to 180 mmol/day. The hypertensive group exhibited decreased (p less than 0.05) dopamine excretion as compared with the normotensive group for both dietary salt intakes. Plasma ANF levels increased (p less than 0.05) in the hypertensive group, but not in the normotensive group, with increasing dietary salt. Plasma aldosterone and urinary norepinephrine and PGE2 were comparable in the two groups for both dietary salt intakes. These data suggest that salt sensitivity is not unique to hypertensive blacks but occurs in normotensive blacks as well. Decreased renal production of dopamine may be a pathogenic factor in the development and maintenance of hypertension in blacks. PMID- 2973443 TI - Mononuclear cell phenotyping in a hemodialysis population. AB - A study of peripheral blood mononuclear cell phenotyping and HIV testing was performed in 39 chronic hemodialysis patients. The following abnormalities were found in comparison to a control population: 1) a lower percentage of lymphocytes in the white blood cell differential count, 2) a lower percentage of total T cells, 3) a lower percentage and absolute number of suppressor T cells and 4) a lower percentage and absolute number of B cells. Both increasing age and blood transfusions were associated with reduced helper cells. PMID- 2973442 TI - Natural killer cells in chronic leukemia. Function and markers. AB - Twenty-two patients with chronic lymphocytic leukemia (CLL) and 14 patients with chronic myelogenous leukemia (CML) were studied with respect to natural killer (NK) cell activity and related cell markers (Leu 7 and Leu 11b). Significantly reduced NK cell activity was detected in peripheral blood from the patients with CLL. Similarly, a reduced number of cells with the markers Leu 7 and Leu 11b (CD 16) were detected in the same patients. Removal of the leukemic cells by centrifugation of cells forming rosettes with mouse erythrocytes led to an augmented, but not fully normalized, NK activity. This indicates that the low NK activity in CLL partly may be due to overgrowth of leukemic cells. However, in spite of the lymphocytic infiltration, the NK cell activity in the bone marrow of CLL patients did not differ significantly from that of normal controls. The patients with CML in the chronic phase as well as patients in the accelerated or blast phase also had a reduced NK activity. The finding that patients in the chronic phase had a reduced NK activity and normal numbers of Leu 11b (CD 16) positive cells, together with no detectable blasts in the peripheral blood, indicates that patients with CML may have an inherent NK cell defect. The highly reduced activity found in patients with the accelerated/blast form may in addition partly be due to overgrowth of leukemic cells. This low NK activity may be of importance in the development of chronic leukemia. PMID- 2973444 TI - Conditional immortalization and/or transformation of rat cells carrying v-abl or EJras oncogene in the presence or absence of glucocorticoid hormone. AB - Early-passaged rat chondroblasts (RX cells) and embryonal fibroblasts (RE cells) are hardly transformed by transfection of activated human H-ras (EJras) or by Abelson murine leukemia virus v-abl oncogene. However, these cells were transformed by v-abl or EJras gene when dexamethasone (DX) was added in the culture medium as well as when co-transfected with retrovirus LTR-linked mouse c myc gene. RX cell lines carrying v-abl (RXabl), RE cell lines carrying v-abl (REabl) and RX cell lines carrying EJras (RXEJ) were established from transformed colonies in the DX-added soft agar. In the absence and in the presence of DX, RXabl cells showed mortal and immortalized, REabl cells showed mortal and transformed, and RXEJ cells showed immortalized and transformed phenotypes, respectively. Especially, immortalization and transformation of REabl1 and REabl3 lines were switched on and off by addition and depletion of DX. v-abl or EJras mRNA levels in tested REabl, RXabl and RXEJ lines cultured without DX was not decreased compared to those cultured with the hormone. The above suggests that, like myc gene, glucocorticoid collaborates with v-abl or activated ras oncogene to transform unestablished rat cells and that the transformation phenotypes were determined not only by the introduced oncogene but by the cellular condition including their tissue origin. Transformation of senescent REabl cells in the absence of DX was tested by transfecting different oncogenes. Among nuclear oncogenes tested, only adenovirus 12 E1A gene could induce transformation of G0 arrested REabl cells in a cooperative fashion with the integrated v-abl gene. PMID- 2973446 TI - Progressive erythrokeratodermia and cochlear hearing impairment. A case report and review of the literature. AB - A female child with congenital progressive erythrokeratodermia combined with sensory hearing loss observed through a period of 5 years is reported. She demonstrates symmetrical hyperkeratotic skin changes, verrucous plaques on her nose cheeks, ears, chin, knees, elbows, and heels. Electron microscopic studies of her skin did not reveal qualitative changes, her moderate to severe hearing impairment is of cochlear origin, moderately progressive, and particularly affecting the high frequencies. Up to now a vascularizing keratitis could not be detected. Her family history is not contributory. Twenty-eight similar cases from the literature, mainly reported as 'KID' syndrome, are reviewed. There are two familial instances. Autosomal dominant inheritance is assumed. We consider the acronymic designation 'KID' syndrome misleading, since the main features of the disorder are a progressive erythrokeratodermia, cochlear deafness, and non obligatory vascularizing keratitis. PMID- 2973445 TI - Alcohol, tobacco, and cannabis use by independently living adults with major disabling conditions. AB - Data from an 18-month prospective study of 205 adults with major disabling conditions revealed that there is a positive but not statistically significant association (p greater than .10) between respondents' scores on a scale which measures both numbers and amounts of substances (alcohol, tobacco, and cannabis) used and bed disability days. There are also nonsignificant positive relationships between scale scores and five of six measures of utilization of medical care. PMID- 2973447 TI - The effect of vanadate upon calcium-stimulated ATPase of the rabbit iris-ciliary body. AB - In many tissues, the level of cytoplasmic calcium mediates cell function. Since cytoplasmic calcium is often maintained at a low level by active calcium extrusion, we examined whether calcium-stimulated ATPase is present in the rabbit ciliary epithelium. A technique was developed to measure calcium-stimulated ATPase in a partially enriched plasma membrane preparation. The enhancement of Na,K-ATPase activity was used to indicate the enrichment of plasma membrane. Marked stimulation of ATPase activity by calcium was observed over a range of calcium concentrations (10(-8) to 10(-3) M). The calcium concentration necessary to elicit half-maximal ATPase activity was 10(-6) M, which is similar to that reported for other membrane preparations. Calcium-stimulated ATPase activity was significantly inhibited in the presence of low concentrations of sodium orthovanadate. The inhibitory influence of vanadate was examined over a range of vanadate concentrations (10(-8) to 10(-3) M). The vanadate concentration needed to produce half-maximal inhibition of calcium-stimulated ATPase was 2 X 10(-6) M. These studies show that calcium-stimulated ATPase inhibition can occur, in vitro, at very low levels of vanadate; it is possible that this might contribute to the chain of events which results in the lowering of aqueous humor secretion reported in vanadate-treated rabbits. PMID- 2973450 TI - Down syndrome. Seasonality and parity effects. PMID- 2973448 TI - Phase II trial of menogaril in advanced colorectal cancer. AB - Menogaril, a new semisynthetic anthracycline antibiotic, was administered to 35 patients with advanced colorectal cancer. The drug was infused over 2 hr at a dose of 160 mg/sqm or 200 mg/sqm repeated every 4 weeks. Twenty-seven patients were evaluable for response and no objective responses were achieved. Myelosuppression, only leukopenia, was usually of mild-moderate degree and occurred in 63% of the patients. Twenty-seven percent of the patients experienced severe leukopenia. Local erythema and phlebitis were frequently observed and were severe in 13% of the patients. Nausea/vomiting (66%) and alopecia (50%) were. of mild-moderate degree. This study suggests that menogaril at these doses and schedule had no activity in advanced colorectal cancer. PMID- 2973449 TI - Treatment strategies after successful thrombolysis in acute myocardial infarction. PMID- 2973452 TI - Persecution. PMID- 2973451 TI - Inhibition of pokeweed mitogen-induced Ig secretion by IgG monoclonal antibodies to MHC class I and class II molecules requires binding of the intact antibody. AB - Seven purified IgG monoclonal antibodies reactive with different epitopes on DQw1, DR, HLA-A3 or p85 glycoprotein of human lymphocytes have each been shown to inhibit pokeweed mitogen (PWM)-induced IgG and IgM secretion in a dose-dependent manner. Binding of these antibodies to their target antigens was required for the suppression. Antibodies of IgG1, IgG2 alpha, and IgG2b subclasses were able to inhibit both IgG and IgM secretion in the PWM system. The mechanisms by which two of the monoclonal antibodies (MoAbs)-77.34, specific for the class II antigen DQw1, and GAP A3, specific for the class I antigen HLA-A3-caused inhibition--were analyzed. The suppressive effects of 77.34 and GAP A3 were maximal when added at the initiation of the culture period. No inhibition of IL-2 production or cellular proliferation was detected. Supernatants obtained from inhibited cultures were not themselves suppressive. The F(ab')2 fragments of either 77.34 or GAP A3 failed to influence PWM-Ig secretion, indicating that intact IgG molecules were required. This suggests that the observed inhibition might be mediated via Fc receptors. Together F(ab')2 fragments of either 77.34 or GAP A3 and a control IgG2a protein did not reconstitute the inhibitory effects of intact 77.34 or GAP A3. Suppression, therefore, required intact Fc portions on the same IgG molecules as those that bound to DQw1 or HLA-A3. These studies suggest that populations of IgG molecules that crosslink sufficient numbers of Fc receptors with other cell surface antigens on peripheral blood mononuclear cells (PBMs) during the early stages of B-cell activation can inhibit Ig secretion. Crosslinking of B-cell Fc receptors with SIg has been proposed by others to act as a negative signal for Ig production; our data raise the possibility that crosslinking of FcR with B-cell plasma membrane components other than SIg can also suppress Ig secretion. PMID- 2973453 TI - Spectinomycin modification. IV. The synthesis of 3' aminomethyldihydrospectinomycins via spectinomycin 3'-cyanohydrins. AB - The C-3'-carbonyl group of N-protected spectinomycin is converted into the corresponding aminomethylalcohols via the intermediacy of cyanohydrins. Methodology for the selective synthesis of either epimer with retention of protection in the aminocyclitol ring provides valuable synthetic intermediates for the preparation of analogs of this important antibiotic. The new methodology provides an efficient synthesis of the highly active 3' aminomethyldihydrospectinomycins. PMID- 2973454 TI - Spectinomycin modification. V. The synthesis and biological activity of spectinomycin analogs with ring-expanded sugars. AB - Tiffeneau-Demjanov rearrangement of 3'-(R)-N,N'-dibenzyloxycarbonyl-3' aminomethyl-dihydrospectinom ycin results in ring expansion affording the homologous analog with a seven-membered sugar ring. In stark contrast, attempted rearrangement of the 3'-S-isomer leads only to epoxide formation. Deprotection of the ring-expanded homolog gives homospectinomycin. The synthesis and biological activity of this interesting new member of the spectinomycin series and the derived dihydrohomospectinomycin is detailed in this paper. PMID- 2973455 TI - Adenylylation of trospectomycin by crude enzyme preparations from Escherichia coli. AB - Employing osmotically shocked lysate of a spectinomycin resistant strain of Escherichia coli, trospectomycin, a new alkylspectinomycin, was adenylylated in the presence of adenosine 5'-triphosphate and magnesium ion. A highly resistant strain of E. coli was obtained by transforming a laboratory strain with a newly constructed plasmid consisting of pBR322 and a determinant for spectinomycin resistance originally found on a low copy number plasmid in E. coli strain NR79. The biologically inactive adenylylated trospectomycin was found to be trospectomycin 6-(5'-adenylate). PMID- 2973456 TI - Human recombinant insulin-like growth factor I. II. Binding characterization and radioreceptor assay development using Balb/c 3T3 mouse embryo fibroblasts. AB - The binding of human recombinant insulin-like growth factor I (IGF-I) to BALB/c 3T3 mouse embryo fibroblasts has been characterized, resulting in the development of a radioreceptor assay. Binding of radioiodinated IGF-I (125I-IGF-I) to washed monolayers of BALB/c 3T3 cells was specific, time dependent, and stable, being maximal after a 10-h incubation at 15 degrees C with no loss of bound ligand or cells through 25 h. Scatchard analysis identified a class of high affinity binding sites with Kd = 59.6 pM and an estimated 1.57 X 10(5) receptors/cell. Half-maximal displacement of bound 125I-IGF-I occurred with 15 to 20 ng/ml unlabeled IGF-I competitor. Insulin-like growth factor II and insulin were far less effective competitors, providing half-maximal displacement at concentrations of 130 to 170 ng/ml and 2 to 3 micrograms/ml, respectively. Epidermal growth factor, transforming growth factor type alpha, and acidic and basic fibroblast growth factors did not compete for 125I-IGF-I binding at 1 microgram/ml. Cells fixed with glutaraldehyde before ligand binding did remain attached to culture dishes more tightly; however such pretreatment destroyed approximately 70% of ligand binding. Crosslinking data indicated that 125I-IGF-I binds specifically to a 330-kDalton receptor as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. This receptor dissociated into 130 kDalton subunits when analyzed in the presence of dithiothreitol. PMID- 2973457 TI - Proteolysis of bacteriophage phi X174 prohead protein gpB by a protease located in the Escherichia coli outer membrane. AB - The gene B protein (gpB) of bacteriophage phi X174 is required for prohead assembly and is removed from prohead during phage maturation. Protease activity was observed in isolated prohead which specifically cleaved gpB. Cleavage of gpB produced two fragments that had apparent molecular weights of 12,300 and 3,700 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino-terminal sequencing of the fragments confirmed that they resulted from the cleavage of gpB and identified the cleavage site as an Arg-Arg at amino acids 76 to 77 of the 120 amino-acid protein. gpB-specific protease activity was observed in both phi X174 infected and uninfected Escherichia coli extracts. This protease activity was localized to the outer-membrane fraction of uninfected cells. Protease activities present in the outer membrane and in isolated prohead produced identical fragments and had the same protease inhibition profile. The gpB-specific activity in uninfected cells was induced by growth at 42 degrees C and was inhibited by the protease inhibitors, 1,10-phenanthroline, EDTA, and N-ethylmaleimide. PMID- 2973458 TI - Genetics and regulation of enterobactin genes in Shigella flexneri. AB - Although Shigella flexneri possesses the genes for two siderophore systems, enterobactin and aerobactin, the enterobactin system is only rarely utilized. To investigate the regulation of enterobactin expression in S. flexneri, all of the genes specifically required for synthesis and transport of enterobactin were cloned from both an expressing (Ent+) and a nonexpressing (Ent-) strain. Notable differences between the cloned genes included endonuclease restriction site changes and the presence of an IS1 element in the Ent- DNA. Southern hybridization revealed that this IS1 element, present at the 3' end of the entF gene, is conserved at this location in different strains and serotypes of Ent- S. flexneri. The Ent- cloned genes were tested for their ability to complement the defect in 11 different Escherichia coli enterobactin mutants. The Ent- genes fully complemented nine mutants but failed to complement the entF mutant AN117 and only partially complemented the entE mutant AN93. Whole-cell RNA isolated from E. coli and the Shigella strains was hybridized to 32P-labeled DNA containing the entB gene or a fragment carrying a portion of the entF gene. E. coli and the Ent+ Shigella strains exhibited derepression of transcription of these genes in low-iron media. Transcription in the Ent- strain remained repressed regardless of iron concentration. Expression of the entB and entF genes was also examined in an Ent- Shigella fur mutant. Expression of entF was only partially derepressed and entB remained fully repressed at all iron concentrations, suggesting that factors other than Fur are responsible for the repression of these enterobactin genes in the Ent- Shigella strains. PMID- 2973459 TI - Characterization of sodium transport in Acholeplasma laidlawii B cells and in lipid vesicles containing purified A. laidlawii (Na+-Mg2+)-ATPase by using nuclear magnetic resonance spectroscopy and 22Na tracer techniques. AB - The active transport of sodium ions in live Acholeplasma laidlawii B cells and in lipid vesicles containing the (Na+-Mg2+)-ATPase from the plasma membrane of this microorganism was studied by 23Na nuclear magnetic resonance spectroscopic and 22Na tracer techniques, respectively. In live A. laidlawii B cells, the transport of sodium was an active process in which metabolic energy was harnessed for the extrusion of sodium ions against a concentration gradient. The process was inhibited by low temperatures and by the formation of gel state lipid in the plasma membrane of this organism. In reconstituted proteoliposomes containing the purified (Na+-Mg2+)-ATPase, the hydrolysis of ATP was accompanied by the transport of sodium ions into the lipid vesicles, and the transport process was impaired by reagents known to inhibit ATPase activity. At the normal growth temperature (37 degrees C), this transport process required a maximum of 1 mol of ATP per mol of sodium ion transported. Together, these results provide direct experimental evidence that the (Na+-Mg2+)-ATPase of the Acholeplasma laidlawii B membrane is the cation pump which maintains the low levels of intracellular sodium characteristic of this microorganism. PMID- 2973460 TI - Isolation of subunits from Methanosarcina barkeri ATPase: nucleotide-binding site in the alpha subunit. AB - The alpha (62,000-dalton) and beta (49,000-dalton) subunits of Methanosarcina barkeri ATPase were purified to homogeneity. The subunits and ATPase complex were trypsinized in the presence of various nucleotides. ATP and ADP changed the trypsin sensitivity of the alpha subunit in the complex and isolated forms, suggesting the presence of a nucleotide-binding site in the alpha subunit. PMID- 2973461 TI - Activation of the erythrocyte Ca2+-ATPase by either self-association or interaction with calmodulin. AB - The octaethyleneglycol mono-n-dodecyl ether solubilized Ca2+-ATPase purified from human erythrocytes has been studied to determine the physical mechanism of its activation by calmodulin. The dependence of Ca2+-ATPase activity on the enzyme concentration shows a transformation from a calmodulin-dependent to a fully active calmodulin-independent form. The transformation is cooperative with a half maximal activation at 10-20 nM enzyme. This suggests that at higher enzyme concentrations interactions between Ca2+-ATPase polypeptide chains substitute for calmodulin-enzyme interactions, resulting in activation. In support of this interpretation, the inclusion of higher octaethyleneglycol mono-n-dodecyl ether concentrations shifts the half-maximal transformation to higher enzyme concentrations. Regardless of the detergent concentration, calmodulin decreases by about 2-fold the enzyme concentration required to observe half-maximal Ca2+ ATPase activation, without affecting the maximal velocity or cooperativity. This indicates that calmodulin facilitates interactions between enzyme molecules. The fluorescein-5'-isothiocyanate-modified Ca2+-ATPase shows an increase in fluorescence polarization which occurs over the same narrow concentration range that is seen with the Ca2+-ATPase activity, confirming association of enzyme molecules. Stimulation of the Ca2+-ATPase activity by calmodulin has revealed a stoichiometry of 0.73, with a dissociation constant of 1.6 nM calmodulin. We have demonstrated by use of calmodulin-Sepharose chromatography that both the calmodulin-dependent and independent Ca2+-ATPase forms bind calmodulin, even though stimulation of activity is seen only with the former one. Our data suggest the following two mechanisms for the Ca2+-ATPase activation: self-association of enzyme molecules or interaction with calmodulin. PMID- 2973462 TI - Troponin T switching in the developing rat heart. AB - A monoclonal antibody specific for cardiac troponin T has been used to investigate troponin changes during development in the rat heart. Specificity of the antibody was determined by immunoblot analysis with purified bovine cardiac troponin. In the rat heart, immunoblot analysis shows that anticardiac troponin T reacts with a 42.5-kDa band in fetal ventricles and with a 41-kDa band in adult ventricles. The faster migrating troponin T is present in traces in the fetal heart and increases markedly during the first 2 weeks after birth, concomitantly with the progressive decrease of the slower migrating form that is no longer detectable in the adult. The pattern of reactivity of the monoclonal antibody is not modified by alkaline phosphatase pretreatment, suggesting that the antibody is not specific for a phosphorylated epitope. Conditions known to affect cardiac myosin composition, such as hypothyroidism and hypertrophy secondary to systemic hypertension, do not change the troponin T isoform profile of adult rat ventricles. The expression and accumulation of the adult isoforms of troponin T are not suppressed by propylthiouracil treatment of pregnant and nursing rats. PMID- 2973463 TI - Insulin and epidermal growth factor stimulate phosphorylation of a 170-kDa protein in intact hepatocytes immunologically related to lipocortin 1. AB - Lipocortin 1 is a steroid-induced, calcium-regulated membrane binding protein (Mr = 39,000) which is a substrate for the epidermal growth factor receptor kinase in intact cells. Using a polyclonal antibody to human recombinant lipocortin 1, we have identified a 170-kDa phosphoprotein in freshly isolated rat hepatocytes which shares antigenic determinants with lipocortin 1. The protein was recognized by four different anti-lipocortin 1 antisera, and antibody binding was inhibited by a 100-fold molar excess of human recombinant lipocortin 1 over antibody. Based on Coomassie Blue staining, the 170-kDa lipocortin-related protein is abundant (approximately 100 ng/10(6) cells) in rat liver, while lipocortin 1 itself is found in very low amounts. Epidermal growth factor and insulin stimulated phosphorylation of this 170-kDa protein in intact rat hepatocytes. The increase in phosphorylation was more pronounced in hepatocytes from dexamethasone-treated animals. The phosphorylation occurred exclusively on serine residues and was maximal 30-60 min after hormone addition. The 170-kDa protein was localized in the cytoplasm in the absence of calcium, while increasing calcium concentration led to partial association with the membrane compartment in rat liver. This 170 kDa protein represents a new member of the class of proteins whose serine phosphorylation is regulated by insulin and EGF and may belong to the family of lipocortin-related molecules. PMID- 2973464 TI - Multiple abnormal beta-hexosaminidase alpha chain mRNAs in a compound heterozygous Ashkenazi Jewish patient with Tay-Sachs disease. AB - Abnormal beta-hexosaminidase alpha chain cDNA clones were isolated from fibroblasts of an Ashkenazi Jewish patient with Tay-Sachs disease. Four abnormal cDNA clones were sequenced in their entirety. We showed previously that three of these mRNAs retained intron 12 with a mutation from G to C at the 5' donor site and that the patient was heterozygous with respect to this splicing defect (Ohno, K., and Suzuki, K., (1988) Biochem. Biophys. Res. Commun. 153, 463-469). One clone retained, in addition to intron 12, intron 13, which was truncated and polyadenylated due to a polyadenylation signal within intron 13. The fourth clone did not contain intron 12 and was missing exon 12. Some of these abnormal mRNAs were also missing one or more of upstream exons. The regions of exon 12-intron 12 and of upstream exons were evaluated in a total of 30 clones, including those completely sequenced, by restriction mapping and Southern analysis with appropriate probes. Of the 25 cDNA clones that included the exon 12-intron 12 region, 11 contained the exon 12-intron 12 sequence with the junctional transversion, and 11 were missing both exon 12 and intron 12. Among the 12 clones that included the region of exon 3-exon 9, 7 were missing one or more of upstream exons. Three clones gave results expected of normal cDNA in the region of exons 12 and 13. One of the three, furthermore, was 3.6-kilobases long and contained the completely normal beta-hexosaminidase alpha chain mRNA sequence on the 3' side and an abnormal 1.7-kilobase segment at the 5' end. These findings suggest that the splicing defect results in either retention of intron 12 or skipping of exon 12 in approximately equal proportions and that remote upstream exons are also frequently excised out. The three clones that were normal in the exon 12 intron 12 region could have derived from the other yet-to-be-characterized mutant allele. However, we were unable to obtain firm evidence that the abnormal upstream sequence is directly related to Tay-Sachs disease. PMID- 2973465 TI - Assessment of the outcome of low back surgery. AB - We aimed to develop a better understanding and method of rating the success or failure of low back surgery by studying 185 patients prospectively. Identical pre operative and postoperative assessment by an independent observer included pain, disability, physical impairment, psychological distress and illness behaviour. Outcome was assessed by the patient, by the observer and by return to work. There was 96% follow-up at two years. Correlation co-efficients varied considerably between the various measures of outcome, both patient and observer appearing to base their assessment mainly on postoperative status rather than on any change produced by surgery. The observer was influenced most by postoperative pain, disability and physical impairment. Patients were influenced most by residual physical impairment, type of surgery and proportional change in disability. Return to work was moderately influenced by postoperative disability and to a larger extent by social and work-related factors. We developed a simple formula to judge overall success or failure which accurately reproduced the combined assessment of patient and observer. If surgical audit is to be meaningful it must be based on an improved understanding of how the outcome of surgery should be assessed. PMID- 2973466 TI - Specific binding of basic fibroblast growth factor to basement membrane-like structures and to purified heparan sulfate proteoglycan of the EHS tumor. AB - The binding of iodinated basic fibroblast growth factor (bFGF) to low-density heparan sulfate proteoglycan purified from the Engelbreth Holm Swarm (EHS) sarcoma was investigated using different techniques. The tumor clearly contained bFGF, the level being comparable to that found in other tissues such as human or bovine brain. 125I bFGF strongly bound to the basement membrane-like matrix of EHS frozen sections as revealed by autoradiography. Iodinated bFGF bound to purified heparan sulfate proteoglycan but not to laminin or collagen type IV, three components isolated from the same tumor. In contrast, acidic fibroblast growth factor (aFGF) displayed negligible binding to heparan sulfate proteoglycan. Binding of bFGF to frozen sections and to purified proteoglycan could be strongly inhibited by heparin and was displaced by an excess of unlabeled factor and completely suppressed after heparitinase and heparinase treatments. Binding was a function of the salt concentration and was abolished at 0.6 M NaCl. Scatchard analysis indicated the affinity site had a Kd of about 30 nM, a value 10-15 higher than that recently reported by Moscatelli (J. Cell. Physiol., 131:123-130, 1987) in the case of the low-affinity binding sites present on the surface of baby hamster kidney (BHK) cells. PMID- 2973467 TI - Differentiation of HL-60 cells is associated with an increase in the 35-kDa protein lipocortin I. AB - Lipocortin I, a 35-kDa protein, has been detected in terminally differentiated monocytes and neutrophils. This calcium-phospholipid binding protein appears to be identical to a 35-kDa protein that can serve as a substrate for the EGF receptor/tyrosine kinase. We have used the human myelocytic cell line HL-60 to explore whether differentiation of hematopoietic cells is associated with changes in the level of lipocortin I. We find that differentiation of HL-60 cells toward the macrophage lineage by the addition of phorbol esters or vitamin D3 or toward neutrophils with dibutyryl cyclic AMP or dimethyl sulfoxide is accompanied by an increase in the cellular content of lipocortin I. In comparison, treatment of HL 60 cells with bryostatin 1, a compound that activates protein kinase C but does not differentiate HL-60 cells, did not effect the level of 35 kDa protein. We have developed a radioimmunoassay to quantitate this protein by using a polyclonal antibody to a synthetic amino terminal peptide of the 35-kDa protein. This antibody recognizes purified pig lung 35-kDa protein as well as a single 35 kDa protein in HL-60 and A-431 cells as determined by Western blotting and immune precipitation. Differentiated HL-60 cells contain 2.6-fold the amount of 35-kDa protein found in undifferentiated HL-60 cells. Our findings that the addition of phorbol esters to HL-60 cells results in an increase in the mRNA for the 35-kDa protein and in an increase in the incorporation of 35S-methionine into the protein suggest that transcriptional activation or increased stability of the mRNA is responsible for the increased rate of synthesis and accumulation of lipocortin I during differentiation of these cells. In the absence of added divalent cations, we have determined that in differentiated HL-60 cells 79% of lipocortin I protein is located in the cytosol while 21% of the total cellular protein is bound to the particulate fraction. The 35-kDa protein can be removed from the particulate fraction by incubation with chelators or treatment with phospholipase A2 or phospholipase C. Addition of the calcium ionophore A23187 to intact differentiated HL-60 cells causes the 35-kDa protein to associate with the particulate fraction of the cell, suggesting that modulation of intracellular calcium levels may play a role in changing the intracellular location of this protein. PMID- 2973468 TI - Receptor-mediated uptake of a mannose-6-phosphate bearing glycoprotein by isolated chicken osteoclasts. AB - We have recently shown that degradation of bone collagen by osteoclasts occurs via proteolytic enzyme activity that depends on an acidic milieu. Since bone resorption occurs in an extracellular, acidic compartment located at the cell matrix attachment site, the osteoclast must deliver the acid collagenolytic enzymes to the cell surface. These observations raise the possibility that the mannose-6-phosphate (M-6-P) receptor, known to sort acidic proteases in other cells, is involved in trafficking lysosomal enzymes to the plasmalemma of bone resorbing cells. To this end we studied receptor-mediated uptake, distribution and release, by isolated chicken osteoclasts, of 125I-hexosaminidase, a M-6-P bearing enzyme. We found that at 4 degrees C, the bone-resorbing polykaryons bind approximately 10,000 molecules of radioligand/cell with a Kd of 0.7 nM, which is endocytosed by osteoclasts at 37 degrees C by a calcium-independent process. Furthermore, 125I-hexosaminidase uptake is unaffected by mannosylated albumin, documenting specificity of the receptor-mediated event. Release of endocytosed enzyme from the cell is also much more rapid than its degradation, attesting to a pathway of uptake and secretion. By autoradiography, the M-6-P bearing ligand is concentrated at the site of osteoclast-bone attachment. Thus, osteoclasts also have the capacity to deliver M-6-P bearing degradative enzymes to their surface at the site of matrix degradation. PMID- 2973469 TI - The use of technetium-99m-HM-PAO in the assessment of patients with dementia and other neuropsychiatric conditions. AB - One hundred fourteen patients suffering from neuropsychiatric conditions have been studied using 99mTc-labeled hexamethylpropyleneamine oxime (HM-PAO) and single photon emission computed tomography (SPECT). Ninety-one patients had a firm clinical diagnosis while 23 were examined without knowledge of the clinical diagnosis. Of the 91 patients, 51 were suffering from dementia, 25 multi-infarct type and 26 Alzheimer's disease. In 19 of the Alzheimer's patients, a characteristic pattern of decreased perfusion in the parieto-occipital regions was demonstrated while those with multi-infarct type showed varying degrees of irregular uptake in the cerebral cortex. These appearances are similar to those shown with positron emission tomography (PET) and we believe that HM-PAO will provide a widely available method for identifying patients with Alzheimer's disease. Twenty-nine patients were suffering from diseases involving the basal ganglia. Fifteen patients with Parkinson's disease showed no significant abnormality in basal ganglia uptake, while 7 or 8 patients with Huntington's disease who had full examinations showed decreased uptake in the caudate nuclei. Similarly, four of six patients with other basal ganglia diseases showed impaired uptake by basal ganglia, and it is concluded that HM-PAO may be useful for the diagnosis and management of this type of patient. Twenty-three patients received HM-PAO imaging as part of their diagnostic work-up; in 19 of them, detailed follow-up was obtained, which indicated that in 7 cases the result of the HM-PAO scan altered the clinical diagnosis and in 9 cases resulted in a change in management. In the remaining 13 cases, the study was found to be helpful in confirming the diagnosis. PMID- 2973470 TI - Angioplasty for peripheral vascular disease. AB - Surgery is no longer the sole option for patients whose peripheral vascular disease requires treatment beyond conservative management; advances in angioplasty have allowed the physician to offer revascularization procedures even to patients who previously would not have been candidates for surgical reconstruction. Several issues, including the location, severity, and number of lesions and the feasibility of surgery, need to be considered in each case. A team approach, utilizing the skills and expertise of the vascular surgeon, the vascular radiologist (or other physician skilled in angioplasty), and the patient's physician, should ensure that the best choice is made for each patient. PMID- 2973471 TI - Atrial natriuretic hormone has biological effects in man at physiological plasma concentrations. AB - Whether atrial natriuretic hormone (ANH) has biological effects at physiological plasma levels in man is not known. Accordingly, we investigated the effects of a 3-h low dose infusion of human ANF (0.75 pmol/kg.min; i.e. 0.0023 micrograms/kg.min) in six normal men, whose sodium intake was normal while sitting, in a single blind, random order, placebo-controlled study. The ANF infusions induced changes in plasma ANH concentrations entirely within the range for normal subjects. The small increases in plasma ANH values were associated with a significant rise in urinary excretion of sodium, magnesium, calcium, and cGMP. PRA and plasma aldosterone concentrations uniformly decreased to 50% and 64% of placebo values, respectively. Systolic and mean arterial pressures fell significantly from preinfusion values during the ANH infusions. These findings constitute strong evidence that ANH is a hormone of physiological significance in the regulation of body fluid volumes in normal man. PMID- 2973472 TI - Peroperative monitoring of distal transluminal dilatation. PMID- 2973473 TI - Congenital lymphangiectasia with fetal cystic hygroma: report of two cases with coexistent Down's syndrome. PMID- 2973474 TI - Susceptibility testing of Neisseria gonorrhoeae to penicillin and spectinomycin in a diagnostic laboratory. AB - Agar dilution breakpoint susceptibility testing using GC, DST, and proteose agars, was performed on consecutive clinical isolates of non-penicillinase producing Neisseria gonorrhoeae to examine the feasibility of using such a system in a diagnostic laboratory. The incidence and level of resistance to penicillin and spectinomycin was also assessed. On DST medium 93 of 200 (46.5%) of isolates were of intermediate resistance to penicillin (MIC 0.12-0.5 mg/l) and 21 of 200 (10.5%) were resistant to penicillin (MIC greater than or equal to 1.0 mg/l). Ninety two of 200 (46%) of isolates had an MIC to spectinomycin of 32 mg/l on DST agar. Isolates seemed to be more resistant when tested on the two other media. The methods used in this study could be applied in a routine diagnostic laboratory for immediate clinical benefit and long term epidemiological studies. To enable direct comparisons to be made between populations at different centres, however, methods for gonococcal susceptibility testing need to be standardised. PMID- 2973475 TI - Cutaneous manifestations of tetrachlorodibenzo-p-dioxin in children and adolescents. Follow-up 10 years after the Seveso, Italy, accident. AB - After an accident in a chemical plant in Seveso, Italy, on July 10, 1976, 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) spread over a populated area. The event was exceptional because children were also affected and because the contamination took place not only through direct exposure but also through inhalation and the ingestion of contaminated foods, especially fruits and vegetables. This paper illustrates the early dermatologic lesions, the late acneic (chloracne) lesions, and their evolution during a 10-year period. Peculiar cutaneous findings, histologic data, and a comparison with previously reported similar accidents are also included. PMID- 2973476 TI - Ketoconazole 2% cream versus hydrocortisone 1% cream in the treatment of seborrheic dermatitis. A double-blind comparative study. AB - Seventy-two patients with seborrheic dermatitis were treated once daily with 2% ketoconazole cream (n = 36) or 1% hydrocortisone cream (n = 36) on a double-blind basis for 4 weeks. For the global evaluation, no significant difference could be seen between the two groups. The clinical response was 80.5% in the ketoconazole group and 94.4% in the hydrocortisone group. For the different symptoms combined (scaling, redness, itching, and papules), no significant difference was seen between the two groups when the total scores at week 2 and at week 4 were compared with the initial scores. The incidence of side effects in both groups was comparably low. PMID- 2973478 TI - Scrotal dermatitis caused by 5-fluorouracil (Efudex). AB - Four patients are described who experienced an acute episode of eczematous dermatitis of the scrotum where 5-fluorouracil (Efudex) cream had inadvertently been applied in the course of treating warts and keratoses. The reaction was not allergic in origin but resulted from the unique absorptive capabilities of scrotal skin, which permitted entrance and local toxic effects of 5-fluorouracil. PMID- 2973477 TI - Guidelines for prescribing isotretinoin (Accutane) in the treatment of female acne patients of childbearing potential. Acne Subgroup, Task Force on Standards of Care. PMID- 2973479 TI - Effect of induced ventricular tachycardia on atrial natriuretic peptide in humans. AB - The effects of induced sustained ventricular tachycardia on the release of plasma immunoreactive atrial natriuretic peptide were evaluated in 11 adult patients undergoing diagnostic electrophysiologic study. Plasma concentrations of atrial natriuretic peptide withdrawn from the right atrium before and during sustained ventricular tachycardia (mean tachycardia cycle length 320 +/- 68 ms, duration greater than 30 s) were determined by radioimmunoassay. Hemodynamic measurements included phasic femoral artery blood pressure and mean right atrial blood pressure before and during ventricular tachycardia. During ventricular tachycardia, atrial natriuretic peptide increased from 93 +/- 49 pg/ml to 234 +/- 195 pg/ml (p less than 0.05), systolic arterial blood pressure decreased from 120 +/- 16 to 70 +/- 23 mm Hg (p less than 0.001), diastolic arterial blood pressure decreased from 63 +/- 8 to 51 +/- 16 mm Hg (p = NS) and mean right atrial blood pressure increased from 3 +/- 1 to 8 +/- 5 mm Hg (p less than 0.02). In six patients, all hemodynamic variables and the atrial natriuretic peptide were measured during repeated stimulation protocols to investigate the effect of ventricular stimulation for ventricular tachycardia induction on atrial natriuretic factor release. Compared with the values obtained during sinus rhythm, there was no significant increase in atrial natriuretic factor during ventricular stimulation at a cycle length of 600 ms (45 +/- 20 versus 52 +/- 21 pg/ml) or at a cycle length of 400 ms (45 +/- 20 versus 57 +/- 18 pg/ml). No significant linear relation could be found among the changes in mean right atrial pressure, systolic arterial blood pressure and the increase in atrial natriuretic peptide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973480 TI - Current complications of diagnostic and therapeutic cardiac catheterization. AB - Data from 2,883 cardiac catheterizations performed during an 18 month period (from July 1986 through December 1987) were analyzed to assess the current complication profile of diagnostic and therapeutic procedures. Procedures performed during the study period included 1,609 diagnostic catheterizations, 933 percutaneous transluminal coronary angioplasties and 199 percutaneous balloon valvuloplasties. Overall, the mortality rate was 0.28% but ranged from 0.12% for diagnostic catheterizations to 0.3% for coronary angioplasty and 1.5% for balloon valvuloplasty. Emergency cardiac surgery was required in 12 angioplasty patients (1.2%). Cardiac perforation occurred in seven patients (0.2%), of whom six were undergoing valvuloplasty, and five (2.5% of valvuloplasty attempts) required emergency surgery for correction. Local vascular complications requiring operative repair occurred in 1.9% of patients overall, ranging from 1.6% for diagnostic catheterization to 1.5% for angioplasty and 7.5% for valvuloplasty. Although the complication rates for diagnostic catheterization compare favorably with those of previous multicenter registries, current overall complication rates are significantly higher because of the performance of therapeutic procedures with greater intrinsic risk and the inclusion of increasingly aged and acutely ill or unstable patients. PMID- 2973481 TI - Predictors of success for coronary angioplasty performed for acute myocardial infarction. AB - To evaluate the predictors and likelihood of success for coronary angioplasty performed in the setting of acute myocardial infarction, 300 consecutive patients with 321 coronary stenoses were studied retrospectively. Success was defined as final diameter stenosis less than 70% and Thrombolysis in Myocardial Infarction (TIMI) flow grade greater than or equal to 2. Nine clinical variables and 15 angiographic variables were assessed. Seventy-nine percent of patients were men; the mean age was 56 +/- 11 years, and 54% of patients also received thrombolytic therapy. The mean left ventricular ejection fraction was 46 +/- 11%, and 18 patients (6%) were in cardiogenic shock. Angioplasty success in the infarct related artery was achieved in 240 patients (80%). In 177 total occlusions (TIMI flow grade less than or equal to 1), the success rate was 75.7% and success was independently predicted by 1) an ejection fraction greater than 30% (p = 0.001); 2) no arterial bend greater than or equal to 45 degrees at the site of angioplasty (p = 0.008); and 3) no triple vessel disease (p = 0.014). In 144 subtotal occlusions (TIMI flow grade greater than or equal to 2), procedural success was achieved in 84.7% and was predicted by 1) absence of thrombus greater than 5 mm (p = 0.023), and 2) absence of other stenoses greater than or equal to 50% in the same artery (p = 0.043), whereas patency without further emergency intervention was achieved in 71.7% and was predicted only by patient age less than or equal to 60 years and absence of cardiogenic shock.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973482 TI - Balloon angioplasty of aortocoronary saphenous vein bypass grafts: a histopathologic study of six grafts from five patients, with emphasis on restenosis and embolic complications. AB - Among 103 patients undergoing percutaneous transluminal balloon angioplasty of obstructed aortocoronary saphenous vein bypass grafts at the Mayo Clinic, six grafts from 5 patients were available for histopathologic examination. The interval from graft insertion to angioplasty ranged from 5 to 105 months and that from angioplasty to graft excision ranged from 6 h to 24 months. Angioplasty produced intimal fissures in three grafts initially obstructed by intimal fibromuscular proliferation. Healing and restenosis resulted from filling of lacerations with fibrocellular tissue and apparently also from restitution of muscular tone. In two of three grafts initially narrowed by atherosclerosis, balloon angioplasty cause extensive plaque rupture and restenosis resulted from extrusion of plaque debris and secondary luminal thrombosis. In the third graft, angioplasty produced no distinct lesions and late restenosis was due to progressive atherosclerosis of the vein graft. Atheroembolization was observed in both patients with plaque rupture and was associated with reoperation in one and death in the other. In conclusion, the results derived from six saphenous vein bypass grafts subjected to balloon angioplasty indicate that restenosis may result from intimal fibrocellular proliferation, thrombosis, restitution of muscular tone and progressive atherosclerosis. Symptomatic atheroembolization may occur in grafts greater than 1 year old. PMID- 2973484 TI - Prevention of hereditary cardiomyopathy in the Syrian hamster with chronic verapamil therapy. AB - The cardiomyopathic Syrian hamster develops genetically determined cardiac necrosis that invariably leads to premature death from congestive heart failure or arrhythmia. This hamster is a valuable model of human disease because it has many features in common with clinical dilated, congestive cardiomyopathy. Previous studies have shown that therapy for several weeks with the calcium channel blocking drug verapamil or the alpha-1 adrenoceptor blocking drug prazosin can prevent myocardial necrosis due to microvascular spasm. Other investigations have demonstrated the positive effects of verapamil in the early stages of disease. It is not clear, however, whether continued treatment can prevent the long-term expression of the cardiomyopathy or whether the disease is genetically predetermined. To address this question, hamsters were treated with oral verapamil for 7 to 8 months during the necrotizing, compensatory hypertrophy and early failure stages of disease. Analysis of myocardial pathologic and biochemical variables demonstrated that continuously treated animals were generally similar to unaffected control hamsters; discontinuous therapy led to partial protection. These findings demonstrate that virtually complete prevention of this hereditary disease is feasible; these results may have important implications for the treatment of human cardiomyopathy. PMID- 2973485 TI - The European Cooperative Study Group trial of intravenous recombinant tissue-type plasminogen activator (rt-PA) and conservative therapy versus rt-PA and immediate coronary angioplasty. AB - This report describes the organization and results of the European Cooperative Study Group trial of intravenous recombinant tissue-type plasminogen activator (rt-PA) plus conservative therapy versus intravenous rt-PA plus immediate intervention (coronary angiography with a view to angioplasty) in patients with acute myocardial infarction. One hundred eighty-four patients were allocated to noninvasive treatment and 183 to an invasive/intervention policy. Immediate angioplasty was attempted in 168 patients (92%) of the latter group. The trial was terminated after a data review by the ethical monitoring committee showed no benefit in the "invasive" group in terms of enzymatic infarct size or left ventricular function, and a trend (although not statistically significant) toward increased mortality in the intervention group. Analysis of coronary patency and residual stenosis at the time of discharge showed no difference in patency rate, but less residual stenosis in the intervention group. The study differed from the Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) and the Thrombolysis in Myocardial Infarction Phase-IIB (TIMI-IIB) trials in that the philosophy of intervention to produce the earliest and most complete reperfusion resulted in a high incidence of angioplasty procedures in vessels not yet reperfused. The reasons for the failure of this philosophy to show benefit are uncertain, but may include an increased risk of reocclusion after angioplasty or reperfusion damage, or both. PMID- 2973483 TI - Quantification of myocardial infarction during coronary occlusion and myocardial salvage after reperfusion using cardiac imaging with technetium-99m hexakis 2 methoxyisobutyl isonitrile. AB - Myocardial imaging with technetium-99m hexakis 2-methoxyisobutyl isonitrile was investigated as a means to assess myocardial infarct size during coronary occlusion and to quantify the extent of salvaged myocardium after coronary occlusion followed by reperfusion. Open chest dogs underwent either a permanent coronary artery occlusion (Group 1, n = 16) or a 2 h occlusion followed by reperfusion (Group 2, n = 15). Animals in both groups were killed 48 h after occlusion. During coronary occlusion, 23 of the 25 dogs that survived the coronary occlusions had abnormal myocardial scintigrams. The scintigraphic perfusion defect size correlated well with the pathologic infarct size (r = 0.85 and 0.95 by planar and tomographic imaging, respectively). The planar scintigraphic defect size, but not the tomographic defect size, overestimated the pathologic size. The planar scintigraphic defect size observed during coronary occlusion was markedly reduced 48 h after reperfusion (24.8 +/- 12.8% to 10.6 +/- 9.7% of the left ventricle, p less than 0.003). The uptake of technetium-99m hexakis 2-methoxyisobutyl isonitrile in the ischemic myocardium increased significantly 48 h after reperfusion (p less than 0.003) and correlated with the increase in regional myocardial blood flow, as assessed by radioactive microspheres (r = 0.83, p less than 0.01). Thus, myocardial imaging with technetium-99m hexakis 2-methoxyisobutyl isonitrile allows reliable demonstration of the presence of acute infarction, estimation of infarct size and quantification of the extent of salvaged myocardium after coronary reperfusion. PMID- 2973486 TI - Insights derived from the thrombolysis and angioplasty in myocardial infarction (TAMI) trials. AB - In the first three phases of Thrombolysis and Angioplasty in Myocardial Infarction (TAMI) multicenter trials, 708 patients received intravenous recombinant tissue-type plasminogen activator (rt-PA) and underwent detailed assessment of clinical, angiographic and ventriculographic outcomes. The cumulative experience and data base afford the opportunity to address several important questions regarding aggressive therapy of myocardial infarction. These include predictive factors of in-hospital mortality, improvement of left ventricular function and the occurrence of recurrent ischemia. Consideration of practical issues, such as thrombolytic therapy for patients with cardiopulmonary resuscitation or previous stroke, use of coronary artery bypass surgery and the effect of operator experience on angioplasty success rate, has also been made possible. The major accomplishments as well as the deficiencies of the current approach to patient management after thrombolysis are reviewed, and the new TAMI trials currently underway are discussed. PMID- 2973487 TI - Administration of thrombolytic therapy in the community hospital: established principles and unresolved issues. AB - In patients with acute myocardial infarction presenting to community hospitals, thrombolytic therapy should be initiated as rapidly as possible under the supervision of a physician. Paramedic or nurse-initiated pre-hospital therapy is currently investigational. Each hospital must have a detailed evaluation and treatment protocol for acute myocardial infarction that specifies the timetable for patient evaluation, who should or should not receive thrombolytic therapy and the proper dose and mode of administration of the agent or agents to be used. Monitoring after the administration of thrombolytic therapy should focus on arrhythmias, hemodynamic problems, recurrent ischemia and bleeding. The role of early cardiac catheterization to detect patients who have unsuccessful thrombolysis or who require mechanical revascularization procedures is under active investigation. The design of the Thrombolysis and Angioplasty in Acute Myocardial Infarction (TAMI) 5 study, which addresses the role of acute interventional catheterization in the treatment of patients with acute myocardial infarction, is described. PMID- 2973488 TI - The risk of reperfusion strategies in the treatment of patients with acute myocardial infarction. AB - Coronary reperfusion early after the onset of acute myocardial infarction can restore vessel patency, improve ventricular function and reduce short- and long term mortality. A number of reperfusion methods have been used to achieve vessel patency in acute myocardial infarction, but each strategy varies considerably in its risk to the patient. Intravenous thrombolysis is associated with an 8% risk of major or minor bleeding and a less than 0.5% risk of intracranial bleeding. This latter risk has been reported to be 1.5% when 150 rather than 100 mg of intravenous recombinant tissue-type plasminogen activator (rt-PA) is used. Invasive procedures such as cardiac catheterization, coronary angioplasty and intraaortic balloon pumping significantly increased the risk of major bleeding at the vascular entry site (from 20 to 40%). Clinical factors such as older age, female gender, lower body weight and hematologic variables including nadir fibrinogen levels and fibrin degradation products also relate to an increased risk. Coronary angioplasty performed alone without thrombolysis is not associated with an excess risk of bleeding and appears to have complications similar to those reported for angioplasty in patients undergoing elective procedures. Systemic anticoagulation has a low (less than 2%) incidence of bleeding, and serious bleeding is rare. These risks must be considered in the decision to institute a treatment strategy in patients with acute myocardial infarction. PMID- 2973489 TI - Coronary angioplasty in patients with unstable angina pectoris: is there a role for thrombolysis? AB - Management of unstable angina has evolved progressively, and coronary angioplasty has recently been shown to be an effective treatment strategy for unstable angina. However, the procedure-related major complication rate is higher when compared with that for angioplasty in stable angina. The underlying pathophysiology may explain this higher complication rate. Rupture of an atherosclerotic plaque associated with thrombus formation is frequent in the pathogenesis of unstable angina. These processes lead to a critical reduction in myocardial blood supply, and coronary angioplasty may effectively interrupt this process. In contrast, coronary angioplasty itself may cause further injury of the already ulcerated intima, have the potential to intensify the ongoing thrombogenic process and lead to an increased frequency of abrupt closure of the artery during the procedure. Therefore, intracoronary streptokinase was used in the procedure in those patients with abrupt closure of the artery immediately after dilation to attempt to improve the immediate result. Coronary angioplasty was attempted in 200 consecutive patients with unstable angina. Initial success in crossing the obstructed artery was achieved in 196 patients; however, an abrupt closure immediately after dilation occurred in 21 of these patients. Of these 21 patients, 12 were also treated with intracoronary streptokinase, and successful dilation was achieved in 9 patients without evidence of necrosis or the need for emergency bypass surgery. Of the remaining nine patients, four successfully underwent redilation with a larger-sized balloon, four underwent urgent surgery (one death postoperatively) and one was treated conventionally. Final success was achieved in 188 patients (94%) without death, the need for emergency surgery or evidence of myocardial necrosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973490 TI - Zearalenone metabolism and excretion in the rat: effect of different doses. AB - Young female rats were orally dosed with either 1 or 100 mg zearalenone kg-1 body weight; zearalenone and metabolites were measured in a 96-h collection of urine and feces by HPLC analysis. Dose had little effect on metabolites formed, or excretion route. In both treatment groups, about 55% of the oral dose was excreted in the feces, while the urine was also a major route of excretion accounting for 15-20% of the administered dose. Zearalenone and metabolites were excreted mainly in the free form, with the production of alpha-zearalenol, the most potent estrogenic metabolite, being greater than 10% of the zearalenone dose. PMID- 2973492 TI - Dracunculus insignis: experimental infection in the ferret, Mustela putorius furo. AB - The laboratory study of dracunculiasis has suffered from the lack of a suitable, readily available animal model. We have been able to experimentally infect ferrets. Mustela putorius furo, with the North American dracunculid, Dracunculus insignis. Ferrets were inoculated with 75 to 100 infective larvae and were necropsied 90 to 240 days later. Guinea worms were recovered from 10 (56%) of 18 ferrets. A total of 44 worms were recovered, for an average of 4.4 worms per infected ferret. Gravid female worms were recovered as early as 128 days postinoculation. Thirteen (87%) of 15 gravid female worms were recovered from the extremities. Living male worms were recovered at 200 days of age, indicating that not all male worms die shortly after mating. First-stage larvae recovered from gravid females as early as 200 days of age were found to be infective to the copepod, Acanthocyclops vernalis. These observations suggest that the ferret is an excellent laboratory animal for dracunculiasis research. PMID- 2973491 TI - In vivo chronic effect of dimethoate and deltamethrin on rabbits. AB - The in vivo effect of dimethoate and deltamethrin on body and organ weights, serum proteins and on plasma acetylcholinesterase (AChE), aromatic esterase and ATPase were examined in growing male rabbits throughout five months period. Both compounds had no significant effect on body weight; however, adrenal, testis & pituitary weights decreased (P less than 0.01); the liver and spleen weights increased (P less than 0.01) in a dose dependent manner. Serum total proteins and globulin decreased (P less than 0.01) in a dose dependent trend, while serum albumin was not greatly affected. AChE activity was increased (P less than 0.01) after 1 month of treatment with the two doses of dimethoate and deltamethrin; thereafter, AChE activity showed 40% inhibition of the control level. The activity of aromatic esterase increased markedly after the first month, then declined gradually until the fifth month. High dose of dimethoate markedly inhibited this enzyme particularly after the 5th month of treatment. Both doses of deltamethrin increased ATPase activity after the first month of treatment, then the ATPase activity was normal. Dimethoate inhibited ATPase particularly at the end of treatment in a dose dependent manner. PMID- 2973493 TI - Current management of congenital heart disease in patients with Down's syndrome. PMID- 2973494 TI - Phagocytosis of opsonized fluorescent microspheres by human neutrophils. A two color flow cytometric method for the determination of attachment and ingestion. AB - A highly reproducible two-color fluorescence cytometric method is described for determining the amount of receptor-mediated and of non-specific phagocytosis by human neutrophils of polystyrene microspheres that have been covalently coated with C3b, iC3b, IgG, mixtures of these, BSA or human F(ab')2. The method includes a correction for externally bound particles and thus measures net phagocytosis. The method involves the incubation of neutrophils with coated green fluorescent microspheres in buffer alone or with cytochalasin D to inhibit phagocytosis followed by staining the externally bound microspheres with red fluorescent antibodies to the immobilized ligand, and determining the green and red fluorescence in a dual laser fluorescence activated flow cytometer. The red to green fluorescence ratio of the mixtures containing cytochalasin D allows for a correction of the green fluorescence due to externally bound microspheres in the mixtures not containing cytochalasin D. The corrected green intensities thus represent net phagocytosis. The specificity of receptor-mediated phagocytosis was confirmed by inhibition with fluid-phase C3b or iC3b or monoclonal antibodies to the receptors. The method can be applied to the determination of both adherent and suspension phagocytosis and can be used as a general model of the phagocytosis of bacteria by neutrophils. PMID- 2973495 TI - A commercially available serum-free medium appropriate for studies of macrophage functions in vitro. AB - This report presents results using a commercially available serum-free medium, HL 1, to study growth and differentiation of the macrophage-like cell line Mm-1. Mm 1 cells grown in RPMI 1640 plus 5% calf serum were compared with the same cells acclimated to and maintained in HL-1. The two medium formulations are compared for their ability to support replication of the line as well as their ability to support the expression of non-specific esterase production, lysozyme secretion, Fc receptor expression, and phagocytosis of latex beads. The serum-supplemented RPMI promoted growth of Mm-1 cells slightly better than HL-1, but the HL-1 medium consistently supported differentiated functions of the cells 8-10% above the level expressed in serum-supplemented RPMI at the same cell density. Based on these findings and the low endotoxin levels of HL-1, we conclude that the medium is appropriate for refined physiological studies of macrophage functions in this cell line. PMID- 2973496 TI - Keeping the caring in nursing. PMID- 2973497 TI - Tourette syndrome: a bizarre and frightening neurological disorder. AB - Gilles de la Tourette (Tourette Syndrome-TS) is a neurological multiple tic disorder caused by a chemical imbalance in the brain. Symptoms follow a waxing and waning course. TS is not degenerative or fatal, but is lifelong and no cure exists. This article addresses etiology, clinical manifestations, importance of early diagnosis and pharmacological treatment. Nursing interventions for the TS patient will also be discussed. PMID- 2973498 TI - Management of spasticity in spinal cord injury: nursing concerns. AB - Nursing diagnoses are proposed for the management of spasticity in the spinal cord-injured person. The diagnoses, pain and self-care deficit related to spasticity, can guide independent nursing management to reduce nociceptive stimulation and increase self-management of spasticity in spinal cord injury. The Gate Control Theory helps to explain alpha muscle susceptibility to nociceptive stimulation and offers rationale underlying nursing management of spasticity. The Modified Ashworth Scale is used to define, measure and evaluate the effectiveness of management of spasticity. Independent nursing management provides the foundation for spasticity intervention. Other interdisciplinary team interventions, including therapist intervention, pharmacological intervention and invasive procedures supplement the nursing management framework. PMID- 2973499 TI - Patients' recollections of their posttraumatic coma. AB - Patients' recollections of posttraumatic coma as viewed retrospectively were explored in this descriptive study. Fifteen patients from two urban north Texas hospitals who incurred traumatic head injury and associated coma (Glasgow Coma Scale score of 8 or less for at least six hours) were interviewed. A set of open ended questions was developed and utilized in a semi-structured interview with each subject. Data such as age, sex, type of injury, how injury occurred, Glasgow Coma Scale score, and duration of coma were collected. Responses elicited during the interviews were analyzed qualitatively. In this investigation seven subjects reported no recollections of being comatose. Eight were able to relate experiences with three prevailing themes: imprisonment, sensory experiences, and death-like experiences. Four subjects recalled actions perceived as helpful and not helpful while in coma. Recollections of unconsciousness appeared to have been affected by the length of coma but not by the Glasgow Coma Scale score or the type of head injury sustained. These findings reveal patients in coma from a traumatic head injury have a limited degree of awareness of the environment. That awareness should be incorporated into nursing management of the comatose patient in acute and chronic care settings. PMID- 2973500 TI - Comparison of nurse and computer recording of ICP in head injured patients. AB - The importance of intracranial pressure monitoring in management and study of the head-injured patient is clearly recognized by the clinician responsible for intensive care. However, in many institutions studies requiring quantitative measures of ICP in head-injured patients are limited by lack of sophisticated computer monitoring equipment. In this study we tested the ability of the nurse to describe ICP course by manual record and compared these results with an on line computerized ICP monitoring system. The nurse recorded a single "end-hour" value of ICP from the bedside monitor while the computer averaged 720 data samples of ICP during the hour. Our results obtained from five head-injured patients undergoing ICP monitoring showed 55% of the 347 data points had a difference in ICP of 0.01 to 3.0 mm Hg and 38% differed between 3.01 and 6 mm Hg. In comparison, 84% of nurse observations were within 6 mm Hg. Comparison of the temporal course of nurse and computer ICP values combined with the frequency distribution of error data indicates the nurse "end-hour" value is a reasonable estimate of the patient's mean ICP for the entire hour as measured by the computer. Nurses can now ask questions regarding various aspects of a patient's ICP course and compare data with other groups as long as the method of data collection is defined in the same manner. PMID- 2973501 TI - Sexuality and spinal cord injury. AB - Spinal cord injury may have a profound effect on the patient's sexuality. In order to maximize the patient's potential, concern and support from health care professionals is needed. This article reviews normal sexual function and changes occurring following spinal cord injury. The PLISSIT model is described and is used to describe nursing interventions designed to deal with the sexual concerns of spinal cord-injured patients. PMID- 2973502 TI - Syncope: neuroscience nursing assessment based on an understanding of underlying pathophysiological mechanisms. AB - To assist neuroscience nurses in their assessment of the client experiencing syncope, the authors propose use of models derived from the three underlying pathophysiological mechanisms of syncope to guide data collection and documentation. This approach is also useful in educating the client and family, as well as nurses and students about the phenomenon of syncope. PMID- 2973503 TI - The Traumatic Coma Data Bank: a nursing perspective, Part I. AB - The Traumatic Coma Data Bank (TCDB) was a collaborative project undertaken to study the nature and causes of severe head injury, allowing patients similar in age, severity of insults and neurological and physical symptoms to be compared in terms of outcome. Systematic uniform data were collected during the prehospital, acute and rehabilitative phases on 581 patients from 6 centers during the pilot phase of the TCDB, June 1, 1979 through May 31, 1982. The pilot phase successfully determined that a collaborative approach to studying head injury was feasible and additional information gained was incorporated into the main phase April, 1983 through April, 1988. A description of the TCDB population as of this writing (730 patients) is included here. PMID- 2973504 TI - Back injuries in industry. PMID- 2973505 TI - Antidepressant drug treatment for poststroke depression. PMID- 2973506 TI - Paralysis, dysphagia and balance problems associated with stroke. PMID- 2973507 TI - [Post-transfusion graft-versus-host disease as the etiology of post-operative erythroderma following open heart surgery--a report of 2 cases confirmed by HLA typing and skin biopsy]. PMID- 2973508 TI - Prostaglandin E2-mediated inactivation of various killer lineage cells by tumor bearing host macrophages. AB - We have previously reported that natural killer (NK) lineage cells are progressively inactivated during tumor development by prostaglandin E2 (PGE2) secreted by host macrophages; that this facilitates spontaneous tumor metastases, which can be prevented by chronic indomethacin therapy (CIT); and that CIT combined with multiple rounds of interleukin 2 (IL-2) can cure experimental metastases and activate all killer lineage cells in situ including NK cells, lymphokine-activated killer (LAK) cells, and tumoricidal macrophages. The present study tested whether PGE2 secreted by tumor-bearing host macrophages exerts pansuppressor effects against the activation of T cells, NK cells, LAK cells, and tumoricidal macrophages from normal splenic cell populations. Macrophages isolated from CBA mice bearing 21-day intraperitoneal Ehrlich ascites tumors (EAT) or C3H/HeJ mice bearing 21-day subcutaneous T58 mammary adenocarcinomas were added (+/- 10(-5) M indomethacin, or a monoclonal anti-PGE2 ab) to syngeneic splenic lymphocytes to examine the effects on 1) polyclonal activation (3-d 3H thymidine [3H-TdR] uptake) with concanavalin A (Con A); 2) one-way (CBA alpha BALB/C or C3H alpha BALBC) MLR (5-d 3H-TdR uptake) and subsequent CTL generation (tested against 51Cr-labeled Con A blasts of the stimulator phenotype); 3) NK activity (after 24-h co-culture) against YAC-1 targets; 4) generation of LAK cell activity (in the presence of 200 or 2,000 units recombinant IL-2 for 3 or 5 days), tested against NK-sensitive and NK-resistant targets. Similar effects were also noted on the generation of tumoricidal activity in normal splenic macrophages cultured for 3 days in the presence of LPS. Normal splenic macrophages added under the same conditions served as controls. Effects of pure PGE2 or PGF2 alpha (10(-6) M) were also examined on these activation events. Results revealed that tumor-host-derived macrophages (but not normal macrophages) markedly suppressed all these activation events and this suppression was abrogated nearly totally by indomethacin and totally by anti-PGE2 ab, indicating its mediation by PGE2. This finding ran parallel with high levels of PGE2 production by tumor-host-derived but not normal splenic macrophages. Pure PGE2 but not PGF2 alpha mimicked these suppressor effects. While tumoricidal activity was generated in normal macrophages in the presence of LPS, IL-2, or IFN-gamma or their various combinations (which led to further augmentation), these agents required the presence of indomethacin to generate significant killer activity in tumor-host-derived macrophages. macrophages.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2973509 TI - Photoreversal of the ultraviolet radiation-induced disappearance of ATPase positive Langerhans cells in the epidermis of Monodelphis domestica. AB - The present study was undertaken to explore the possible causes of ultraviolet radiation (UVR)-induced disappearance of ATPase-positive, epidermal Langerhans cells (LC). Monodelphis domestica was used because it has the capacity for photoreactivation of UVR-induced pyrimidine dimers in epidermal DNA. Single, 330 J/m2 (ears) or 500 J/m2 (back) UVR exposures (FS-40 sunlamps) reduced the numbers of ATPase-positive epidermal LC in M. domestica ears to approximately 15% of those in unirradiated ears and approximately 37% of those in unirradiated dorsal skin. Immediate 90-minute exposures to photoreactivating light (PRL, 320-400 nm) post-UVR reversed the effects of UVR, resulting in ATPase-positive LC numbers not being significantly different from controls. Exposure to PRL immediately preceeding UVR did not prevent ATPase-positive LC disappearance. The photoreactivation of UVR-induced ATPase-positive LC disappearance indicates that DNA damage (pyrimidine dimers) is involved in the loss of ATPase-positive LC. PMID- 2973510 TI - The presence of atrial natriuretic peptide in canine cerebrospinal fluid and its possible origin in the brain. AB - The presence of atrial natriuretic peptide (ANP) in canine cerebrospinal fluid (CSF) was clearly demonstrated and an attempt was made to determine its origin as either the brain or the atrium. The concentration of ANP in canine CSF was 0.78 +/- 0.37 pmol/l (n = 31) and showed no evident correlation with that in plasma (r = 0.12). Physiological doses of human alpha-ANP (alpha-hANP) were continuously infused intravenously into nine dogs, and ANP concentrations in CSF and plasma were examined six to eight times within a 120-min period following this. The ANP level in CSF was not influenced by the systemic administration of alpha-hANP up to 180 min. Only one low molecular weight peak corresponding to alpha-hANP could be obtained from the CSF samples, while both low and high molecular weight peaks were observed for plasma ANP by gel permeation chromatography. In the atrial and hypothalamic tissue extracts the same kinds of peaks were also evident. These results prove the presence of ANP in canine CSF and that it does not come from blood that has seeped across the blood-CSF barriers, but suggest that it may originate from the brain. PMID- 2973511 TI - Increase in plasma concentrations of atrial natriuretic peptide during infusion of hypertonic saline in conscious newborn calves. AB - Plasma concentrations of atrial natriuretic peptide (ANP), plasma renin activity (PRA), plasma concentrations of aldosterone, urine flow rate and sodium and potassium excretion were studied in two groups of four conscious 3-day-old male calves, infused with hypertonic saline or vehicle. Hypertonic saline infusion (20 mmol NaCl/kg body weight) was accompanied by a progressive rise in plasma concentrations of ANP (from 16.5 +/- 0.2 pmol/l at time 0 to 29.3 +/- 3.0 pmol/l at 30 min; P less than 0.05) and by a gradual decrease in PRA (from 1.61 +/- 0.23 nmol angiotensin I/l per h at time 0 to 0.54 +/- 15 nmol angiotensin I/l per h at 90 min; P less than 0.05); there was no change in the plasma concentration of aldosterone. Within the first 2 h of the 24-h urine collection period there was a marked rise in urine flow rate and sodium excretion in treated calves when compared with control animals (66.0 +/- 8.3 vs 15.9 +/- 1.2 ml/kg body weight per 2 h (P less than 0.05) and 6.7 +/- 1.3 vs 0.4 +/- 0.02 mmol/kg body weight per 2 h (P less than 0.01) respectively). During the following 22 h, urinary water and sodium excretion remained at significantly high levels. Thus, in the conscious newborn calf, changes in plasma ANP levels and urinary water and sodium excretion during hypertonic saline infusion are compatible with the hypothesis that endogenous ANP participates, at least in part, in the immediate diuretic and natriuretic renal response induced by the sodium overload. PMID- 2973512 TI - Return to work certification. PMID- 2973513 TI - Clinical predictors of outcome of acute episodes of low back pain. AB - In this prospective study, predictors of outcome were identified for patients (n = 116) who presented to their family physician with acute mechanical low back pain. Short-term outcome was measured by the number of days lost from work and longer term outcome was measured by disability at the six-week follow-up. Unlike other published work, this study did not find obesity or a history of previous back problems to be related to a poorer outcome from acute episodes of low back pain. Among those patients not involved in manual labor, a history of anxiety or depression was a significant predictor of both greater work loss and longer term disability. Among this same group, cigarette smoking was also found to be related to greater long-term disability from acute low back pain. Further study of this relationship is needed. The number of hours of manual labor performed daily was a strong predictor of poor outcome (both short- and long-term) of acute episodes of low back pain. Among both manual laborers and professional-technical workers, the number of days off work (at bed rest) prescribed by the physician was significantly related to greater absenteeism from work; the physician's diagnosis of an actual or possible disc problem was also related (P less than .05) to greater work loss among manual laborers. Neither of these factors, however, was related to longer term disability. PMID- 2973514 TI - The simultaneous phenotyping of erythrocyte acid phosphatase, esterase D, phosphoglucomutase and adenosine deaminase by isoelectric focusing. PMID- 2973515 TI - Adult onset motor neuronopathy in the juvenile type of hexosaminidase A and B deficiency. AB - Two sisters presented with progressive muscle cramps, as well as wasting and weakness of the legs with onset after age 20. They also showed intention tremor of the upper extremities and dysarthria starting during the first decade. The older patient also had fasciculations; the younger, hyperreflexia. Total plasma beta-hexosaminidase (Hex) activity with 4-methylumbelliferyl-acetyl-glucosamine as substrate was reduced to 1.4% and 2.7% of the control in the 2 patients, respectively. Hex A activity measured by 4-methylumbelliferyl-N-acetylglucosamine 6-O-sulphate as substrate was 9.9% and 12.8% of the mean control value in the 2 patients, respectively. Hex B activity was undetectable in both patients. Leukocyte total Hex activity was 7-8% of normal; residual Hex A activity in the 2 patients was 17.8% and 16.3% of normal controls, respectively. Fibroblastic residual Hex A activity in the 2 patients was 9.6% and 22% of normal mean value, respectively. Appendiceal ganglion cells contained membranous cytoplasmic bodies in the younger patient. Thin layer chromatography of the appendiceal extract from one patient (III/2) showed a marked increase of GM2 ganglioside, and some increase of GM3 ganglioside. Northern blots performed on fibroblast cell lines from both patients for the demonstration of alpha and beta locus messenger RNA showed no difference between patients and control. These patients have a rare form of adult-onset progressive motor neuron disease presumably due to abnormal beta subunits, causing severe deficiency of both Hex A and Hex B. The phenotypic expression of this disease is similar to motor neuron disease due to alpha locus mutations, which suggests that the Hex A deficiency, even though only a partial one, may be the important pathogenic factor. PMID- 2973516 TI - Fine structural localization of Ca2+-ATPase activity at the frog neuromuscular junction. AB - Ca2+-ATPase activity has been shown to be associated with the nerve terminal plasma membrane at the frog neuromuscular junction. Using a modification of the Wachstein-Meisel procedure for localization of phosphatases, a dense reaction product forms at the neuronal plasma membrane/Schwann cell interface. It has been determined that this reaction product is associated with the plasma membrane of the nerve terminal and not the plasma membrane of the Schwann cell. No ATPase activity is demonstrated at the presynaptic portion of the plasma membrane facing the synaptic gap. When a preparation is denervated, a Schwann cell process moves into the space previously occupied by the nerve. There is no ATPase activity associated with the Schwann cell plasma membrane. Conversely, when the Schwann cell is selectively injured, dense reaction product continues to be associated with the nerve terminal plasma membrane. There is some indication that this ATPase activity is dependent on the presence of Ca2+ and Mg2+. Incubation in the calmodulin inhibitor, R24571, shows little inhibition of labelling. PMID- 2973517 TI - Quantitation of renal function with technetium-99m MAG3. AB - The technetium-labeled hippuran analog [99mTc]MAG3 was compared with [131I]hippuran in 50 patients using a quantitative renal function protocol that includes: (a) estimation of effective renal plasma flow by a single-injection, single-sample plasma clearance method, (b) determination of relative function of right and left kidney from the initial count rate over each kidney, and (c) comparison of recovered urine activity with plasma disappearance. This protocol is suitable for routine clinical use, and, in fact, has been used heavily at our clinic for a number of years. By slight modification of the formulas, the results obtained with [99mTc]MAG3 agreed well with those using [131I]hippuran. We conclude that [99mTc]MAG3 can be substituted for [131I]hippuran in the quantitative protocol, with the better image quality and lower radiation dose (in abnormals) of a technetium-labeled agent. PMID- 2973518 TI - The effect of dietary essential fatty acid deficiency on the composition and properties of the liver microsomal membrane of rats. AB - The effect of dietary essential fatty acid (EFA) deficiency on lipid composition, fluidity and important enzyme and transport activities of liver microsomal membrane was studied in weanling rats. After 133 d of EFA deficiency, no difference was noticed in membrane phospholipid, cholesterol and protein levels, but a significant change occurred in the fatty acid composition of bilayer phospholipids. In EFA-deficient rats, linoleic (18:2(n-6] and arachidonic (20:4(n 6] acids were both severely lower while oleic (18:1(n-9], palmitoleic (16:1(n-7] and particularly 5,8,11-eicosatrienoic (20:3(n-9] acids were significantly higher than in controls. The higher level of the latter tended to compensate for the lower level of 20:4(n-6). Membrane fluidity, as estimated by the reciprocal of the order parameter S, was lower in the deficient rats than in the controls, and all the measured microsomal enzyme activities were markedly affected. NADH-Cyt b5 electron transferring system, coupled with the fatty acid desaturation system, was higher than in controls. In contrast, the cytochrome P450 complex activity was lower and some of the important liver detoxifying enzyme activities were lower due to physical-chemical changes in the microsomal membrane. Calcium uptake and Ca2+-ATPase activity were also significantly lower in EFA-deficient rats than in controls. It was concluded that fatty acid composition may be the major factor contributing to membrane fluidity and function and that EFA might play a role in regulating the intrinsic membrane protein activities. PMID- 2973519 TI - Glycosaminoglycan changes in submandibular glands of experimental diabetic rats. PMID- 2973520 TI - Biochemical aspects of collagen in submandibular glands of Streptozotocin-induced diabetic rats. PMID- 2973521 TI - Induction of macrophage motility by a T-cell line from Balb/c mice specific for Plasmodium berghei malaria. AB - A long-term antimalaria T-cell line (AMTL) expressing a helper phenotype (Thy 1.2+, Lyt 2.2-) was established from Plasmodium berghei-recovered Balb/c mice. The ability of this T-line to induce macrophage motility was measured in vivo and in vitro. Adoptive transfer of AMTL cells to normal Balb/c mice showed an increased delayed hypersensitivity response to the homologous antigen, i.e., parasitized erythrocytes (PE). In vitro, AMTL culture supernatant (AMTL-SUP) augmented chemotactic locomotion of macrophages derived from both normal and infected mice. However, the effect on normal macrophages was significantly higher. AMTL cells adoptively transferred to normal mice had no effect on parasitemia levels or mortality rate after subsequent infection with P. berghei. Partial characterization of the AMTL-SUP indicated the involvement of a protein of about 12,600 Daltons in the enhancement of chemotaxis. These findings suggest that the AMTL cells and chemoattractants produced by them can induce macrophage motility, and that the macrophage malfunction in Balb/c with P. berghei infection is not due to defects at the T-lymphocyte level. PMID- 2973523 TI - An enhanced standard computer keyboard system for single-finger and typing-stick typing. AB - A prototype keyboard system was developed, using off-the-shelf hardware and software, as an inexpensive keyboard-based system to facilitate data entry for single-finger and typing-stick typists. Evaluation established that the system can increase entry rate by 50 percent or more. The underlying concepts may provide a basis for developing other configurations that accelerate and simplify computer keyboard use for persons with a variety of hand impairments. PMID- 2973522 TI - Taenia taeniaeformis: cellular reconstruction of athymic mice and role of L3T4+ helper T lymphocytes in the early infection. AB - The role of T helper lymphocytes (L3T4+) in the early response to Taenia taeniaeformis metacestodes was investigated. Athymic BALB/c-nu/nu mice (susceptible) were inoculated intraperitoneally with the following cell populations from congenic BALB/c-nu+ + mice (resistant): (a) whole spleen single cells, (b) thymus single cell suspensions, or (c) spleen cells pretreated with anti-L3T4 monoclonal antibody before the injection. The mice were given 3 weekly injections of cells and then infected orally with 300 eggs 7 days after the last injection. Cryostat sections of the liver from the infected mice were examined at 6 days postinfection (PI) for parasite viability, the numbers of eosinophils, and L3T4+ T lymphocytes present within 100 micron of the parasite and for the presence of biotin in hepatocytes (involved in biosynthesis of fatty acids) around the parasite. The success of the cellular reconstitution of athymic mice with the lymphoid cells was measured by a T-cell mitogenic assay with concanavalin A (ConA). The cellular reconstitution of athymic mice with a mixture of lymphoid cells from the spleen and thymus of BALB/c-nu/ + mice resulted in both parasite death and eosinophil infiltration. Reconstitution with mature splenic cells alone resulted in a greater parasite killing and eosinophil infiltration as compared to reconstitution with thymic cells. The better reconstitution with splenic cells was reflected in a greater mitogenic response to ConA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973525 TI - Laparoscopic and vaginal colpotomy for the excision of infiltrating cul-de-sac endometriosis. AB - Palpable endometriotic nodules deep in the cul-de-sac and vagina represent the extension of intraperitoneal disease. Although such nodules used to be excised with vaginal colpotomy and by tracing the endometriosis to the peritoneum, the dissection of these lesions under laparoscopic visualization had aided in their removal. Of seven patients who were approached with a plan for combined laparoscopic and vaginal excision, five underwent the procedure. The last two required laparotomy due to bowel muscularis involvement. PMID- 2973524 TI - [Fibromuscular dysplasia of the external iliac artery. Treatment by endoluminal angioplasty]. AB - Fibromuscular dysplasia of external iliac artery in a 30 year old woman was treated by endoluminal angioplasty with excellent results: claudication symptoms were absent during a 6 month follow up. PMID- 2973526 TI - Proceedings of the 16th annual meeting of the American Association of Gynecologic Laparoscopists. San Francisco, California, November 1987. PMID- 2973527 TI - Enhancement of fluorinated pyrimidine-induced cytotoxicity by leucovorin in human colorectal carcinoma cell lines. AB - Reduced folates have been shown to increase the cytotoxicity of 5-fluorouracil (5 FU) by stabilizing the 5-fluoro-2'-deoxyuridine-5'-monophosphate-thymidylate synthase complex, thus increasing the block in the DNA synthetic pathway. Using an in vitro colorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cytotoxicity assay, we tested the effects of 5-FU and 5-fluoro-2' deoxyuridine (FdUrd) with and without leucovorin (LV) on a panel of 11 human colorectal carcinoma cell lines. The effect of LV on 5-FU and FdUrd was quantitatively similar. A clinically achievable level of LV (20 microM) increased the cytotoxicity in all three replicate experiments in 10 of the 11 cell lines (P less than .05, binomial test). LV alone at a concentration of 20 microM had no effect on cell survival. In three cell lines, 50% inhibition of growth occurred at a clinically achievable area under the curve of 5-FU alone. With the addition of LV, one additional cell line showed 50% growth inhibition at a clinically achievable level of 5-FU. Hence large clinical trials may be necessary to detect a significant improvement in survival as a result of adding LV to the fluorinated pyrimidines. PMID- 2973529 TI - The combination of cyproterone acetate and low dose diethylstilbestrol in the treatment of advanced prostatic carcinoma. AB - Cyproterone acetate is a steroidal antiandrogen with weak progestational activity that results in the partial suppression of pituitary gonadotropin. We demonstrate that the associated decrease in serum testosterone is more complete and prolonged if cyproterone acetate (200 mg. daily) is combined with a low dose of diethylstilbestrol (0.1 mg. daily). The effectiveness of the combination in the treatment of prostatic carcinoma was investigated in 51 patients with stage D2 malignancy. From an initial concentration of 360 +/- 23 ng. per dl. (mean +/- standard error), serum testosterone decreased to 56 +/- 5 ng. per dl. after 1 week and reached a plateau of approximately 30 ng. per dl. after 2 months. This was accompanied by a decrease in serum prostatic acid phosphatase from a mean starting concentration of 43 +/- 11 ng. per ml. to a low of 3 +/- 1 ng. per ml. at 12 months; the proportion of normal values increased from 10 to 80 per cent. A complete response was observed in 7 patients (13 per cent), partial response in 35 (69 per cent) and stable disease in 8 (16 per cent), yielding an over-all objective response rate of 98 per cent (1980 National Prostatic Cancer Project criteria). The actuarial median time to progression was 17 months and the median survival time was 23.5 months. In 26 patients who subsequently had signs of progressive carcinoma the relapse rate in bone (85 per cent) far exceeded that in nonskeletal sites (23 per cent). The incidence of cardiovascular toxicity was 12 per cent. These results indicate that cyproterone acetate and low dose diethylstilbestrol may be co-administered to achieve a synergistic and potent androgen withdrawal effect in the treatment of advanced prostatic carcinoma. PMID- 2973528 TI - Mechanism of tumor destruction following photodynamic therapy with hematoporphyrin derivative, chlorin, and phthalocyanine. AB - The effect of photodynamic therapy on the tumor microvasculature in the first few hours after treatment was studied at the light and electron microscopy levels. BALB/c mice with EMT-6 tumor received ip injections of hematoporphyrin derivative, chlorin, or phthalocyanine, and 24 hours later, the tumors were treated with light at 100 J/cm2 at the appropriate therapeutic wavelength for each photosensitizer. Animals were killed and their tumors removed at time 0, 30 minutes, 1 hour, and 2, 4, 6, 8, 12, 16, and 24 hours after treatment. The results indicate that for all three sensitizers the effects of photodynamic therapy leading to rapid necrosis of tumor tissue are not the result of direct tumor cell kill but are secondary to destruction of the tumor microvasculature. The first observable signs of destruction occur in the subendothelial zone of the tumor capillary wall. This zone, composed of dense collagen fibers and other connective tissue elements, is destroyed in the first few hours after phototherapy. However, the ultrastructural changes seen in this zone are different for the hematoporphyrin derivative, compared with chlorin and phthalocyanine. Binding of photosensitizers to the elements in this zone as well as altered permeability and transport through the endothelial cell layer because of the increased intraluminal pressure may be key features of tumor destruction. PMID- 2973530 TI - Re: Cardiovascular side effects of diethylstilbestrol, cyproterone acetate, medroxyprogesterone acetate and estramustine phosphate used for the treatment of advanced prostatic cancer: results from European Organization for Research on Treatment of Cancer Trials 30761 and 30762. PMID- 2973532 TI - [Effect of infusion of atrial natriuretic peptide on endocrine function after administration of endotoxin in dogs]. PMID- 2973533 TI - [Clinical evaluation of repeat-PTA for post-PTA re-stenosis]. PMID- 2973531 TI - Comparative immunogenicity of plasma and recombinant hepatitis B virus vaccines in homosexual men. AB - A randomized, double-blind clinical trial of plasma-derived and DNA recombinant hepatitis B virus vaccines was conducted in 186 homosexual men. Nine months after the immunization series (three doses) began, the seroconversion rate in the plasma vaccine group was 88% (68/77); this was significantly higher than the 74% (60/81) response rate of the recombinant vaccine group. Men positive for antibody to the human immunodeficiency virus (HIV) had a considerably higher nonresponse rate to either vaccine than expected in non-HIV-infected homosexual men. The odds ratios of nonresponse to hepatitis B virus vaccine for HIV-seropositive vs HIV seronegative subjects were 12.0 (95% confidence interval, 1.7 to 89.3) and 13.6 (95% confidence interval, 2.3 to 148.3) for the plasma and DNA recombinant vaccines, respectively. PMID- 2973534 TI - [Quantitative estimation of myocardial thickness by the wall thickness map with Tl-201 myocardium SPECT and its clinical use]. PMID- 2973535 TI - [Re-evaluation of the approach to the problem of physically handicapped children]. PMID- 2973536 TI - A medical accident compensation system: a model act. PMID- 2973537 TI - [Plastic repair of the anterior abdominal wall in postoperative and recurrent hernia]. PMID- 2973538 TI - [Rendering aid to the victims of abdominal trauma at a district hospital]. PMID- 2973539 TI - [Clinical characteristics of dysentery today]. PMID- 2973540 TI - [Comparative effectiveness of non-drug and drug methods of treating patients with hypertension at a sanatorium-preventorium]. PMID- 2973541 TI - Variation in T helper cell/T cytotoxic-suppressor cell index during cardiac operations. AB - After surgical procedures, humoral and cell-mediated immunity responses decrease. Both anesthesia and surgical trauma play an important role in this effect. Cardiac operations induce a greater decrease in immunologic response than other surgical operations. On the other hand, identification of functional lymphocyte subclasses by means of appropriate monoclonal antibodies appears to provide an accurate measurement of cellular immune competence. Employing these methods, we found a significant decrease of T helper cell/T cytotoxic-suppressor cell ratio in the periperal blood of 20 patients undergoing cardiac operations. This decrease during the period of anesthesia (before surgical incision) is due as much to a decrease of T helper cell percentage as to an increase of T cytotoxic cell percentage. However, during the surgical trauma period (surgical incision to the end of the operation) it is due to a greater increase of T cytotoxic suppressor cell percentage without significant changes in the percentage of T helper cells. PMID- 2973542 TI - Differentiating human leukemia cells express heparanase that degrades heparan sulfate in subendothelial extracellular matrix. AB - Human promyelocytic (HL-60) and monoblast-like (U-937) leukemia cell lines were tested for expression of an endoglycosidase (heparanase) capable of degrading heparan sulfate (HS) side chains in the subendothelial extracellular matrix (ECM). Heparanase activity has been previously shown to be expressed by activated lymphocytes and macrophages and by highly metastatic tumor cells, in correlation with their ability to invade blood vessels and extracellular matrices. Incubation of HL-60 and U-937 cells with sulfate-labeled ECM in the presence of 12-O tetradecanoyl-phorbol-13-acetate (TPA) resulted in heparanase-mediated release of heparan sulfate degradation products. This degradation was inhibited by heparin, stimulated by plasminogen and not expressed by cells treated with retinoic acid or dimethylsulfoxide and undergoing neutrophilic differentiation. Heparanase activity was not detected in media conditioned by HL-60 and U-937 cells but was found in their cell lysates, regardless of whether or not the cells were exposed to TPA. These findings imply that TPA-induced differentiation of human myeloid leukemic cells to macrophage-like cells, but not to neutrophilic granulocytes, is associated with expression on the cell surface of a preformed heparanase activity. The enzyme may serve as a marker for human cell differentiation into macrophages, allowing the differentiating cells to traverse the vascular compartment and reach their target sites. PMID- 2973543 TI - [Local fibrinolysis and percutaneous transluminal angioplasty in embolism of the renal artery]. PMID- 2973544 TI - Presence of the atrial natriuretic factor (ANF) in human ascitic fluid. AB - Presence of atrial natriuretic factor (ANF)-like material was demonstrated by radioimmunoassay in ascitic fluid of 14 patients with cirrhosis of the liver. Immunoreactive ANF concentrations (M +/- SEM) were 2.4 +/- 0.5 fmol/ml in ascites, significantly lower (p less than 0.001) than the corresponding plasma concentrations of 15.5 +/- 2.6 fmol/ml. High performance gel permeation chromatography and reverse phase high performance chromatography of the ascitic ANF immunoreactivity showed correspondence to the alpha human ANF (99-126). ANF levels in ascites were significantly (p less than 0.01) correlated to levels in plasma (r = 0.66). PMID- 2973546 TI - Stress management training for parents of children with severe handicaps. PMID- 2973545 TI - Age-related changes of methionine-enkephalin and beta-endorphin/beta-lipotropin immunoreactivity in human CSF. AB - Methionine-enkephalin (ME-IR) and beta-endorphin (BE-IR) immunoreactive material CSF concentrations have been measured in subjects of different ages affected by lumbar or cervical disk hernia. The two peptides exhibited different age-related trends. ME-IR levels rose significantly with age while no changes were observed in the case of BE-IR. PMID- 2973547 TI - Physical educators' attitudes toward teaching students with handicaps. PMID- 2973548 TI - Dental treatment program for patients with mental retardation. PMID- 2973549 TI - Maternal hyperglycemia in pregnant rats: its effect on growth and carbohydrate metabolism in the offspring. AB - Mild hyperglycemia during the last half of pregnancy was achieved by administration of streptozotocin to pregnant rats on the fifth day of gestation. Citrate buffer (vehicle for streptozotocin) was administered to control rats also on the fifth day of gestation. The pups born to the streptozotocin-treated mothers had higher birth weight, pancreatic insulin content, plasma insulin, and C-peptide concentrations compared with pups born to control mothers. The plasma glucose concentrations of the pups were similar between the two groups. The pups who were identified as macrosomic (birth weight greater than 1.7 SD than the mean of the control pups) maintained an accelerated postnatal growth through the first 10 weeks of age in female rats and in the first 3, and at 5 and 6 weeks of age for the male rats. The accelerated growth in the female rats was associated with higher perirenal-ovarian and salpingeal fat weight at 6 weeks. At 10 weeks of age, higher plasma insulin and glucose concentrations were observed following oral glucose challenge in both male and female macrosomic rats than in the control rats. At 12 weeks of age, only the female macrosomic rats showed abnormal glucose response due to peripheral insulin resistance. In the male rats at 12 weeks of age, a higher plasma insulin concentration in the macrosomic group was associated with a normal plasma glucose response to oral glucose challenge. We conclude that mild maternal hyperglycemia in rats resulted in fetal hyperinsulinemia and accelerated fetal growth.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973550 TI - [A species of malarial mosquito new to the fauna of the USSR--Anopheles (Cellia) multicolor Camb. (Diptera: Culicidae)]. PMID- 2973551 TI - [Vectors of the causative agents of leishmaniasis in Chimkent Province, the Kazakh SSR]. PMID- 2973552 TI - [Data from a study of a natural focus of Marseilles fever in one of the Crimean districts over a 40-year period (1947-1986)]. PMID- 2973553 TI - [Detection of Fasciola in deceased persons in Samarkand Province]. PMID- 2973554 TI - [Serologic examination of convalescents and members of their families in tick borne Central Asian borreliosis]. PMID- 2973555 TI - [A case of finding Dirofilaria repens in an inhabitant of Turkmenia]. PMID- 2973556 TI - Preliminary report: 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure to humans- Seveso, Italy. PMID- 2973557 TI - Cerebral metabolism of glucose in benign hereditary chorea. AB - Benign hereditary chorea (BHC) is an autosomal dominant disorder characterized by chorea of early onset with little or no progression. There is marked clinical variability in this disease with some subjects having onset in infancy and others with onset in early adulthood. In contrast to Huntington's disease (HD), there is no dementia. Computed tomography is normal in all subjects with no evidence of caudate nucleus atrophy. We present the results of positron emission tomography using 18F-2-fluorodeoxyglucose on three patients with this disorder from two families. Cerebral glucose metabolism in one patient was decreased in the caudate nucleus, as previously reported in HD. The other two persons from a second family showed a relative decrease in metabolic rates of glucose in the caudate when compared with the thalamus. It appears that caudate hypometabolism is not specific for HD. These findings suggest that the caudate nucleus may play a significant role in the pathophysiology of some persons with BHC. PMID- 2973558 TI - Ipsilateral blepharospasm and contralateral hemidystonia and parkinsonism in a patient with a unilateral rostral brainstem-thalamic lesion: structural and functional abnormalities studied with CT, MRI, and PET scanning. AB - A patient developed progressive right hemidystonia in childhood. Subsequently, left-sided blepharospasm, slurred and stuttering speech, and right-sided rigidity and bradykinesia, responsive to dopamine agonists, appeared. Investigation with computed tomography and magnetic resonance imaging (MRI) at age 43 years revealed a left-sided calcified rostral brainstem-thalamic lesion of uncertain aetiology. Although no structural lesion was seen in the striatal regions, L-[18F] fluorodopa uptake was severely diminished in the left striatum but normal on the right. Dopamine receptor binding identified by [11C]-methylspiperone was in the normal range on both sides. PMID- 2973560 TI - Cerebrospinal fluid GABA and homocarnosine concentrations in patients with Friedreich's ataxia, Parkinson's disease, and Huntington's chorea. AB - Free and total gamma-aminobutyric acid (GABA) and homocarnosine concentrations were measured in the lumbar cerebrospinal fluid (CSF) of patients with Friedreich's ataxia, Huntington's chorea, and Parkinson's disease (with and without levodopa treatment), and compared with those determined in control subjects. Values found in Friedreich's ataxia or Parkinson's disease were not significantly different from those in controls. Unexpectedly, in Huntington patients, known to have a characteristic decrease in GABA concentrations in specific brain areas, CSF concentrations of total GABA and homocarnosine were significantly higher, whereas free GABA was not different from controls. These findings indicate that the measurement of CSF GABA and homocarnosine in patients with CNS degenerative diseases should be interpreted cautiously. PMID- 2973561 TI - Genetic prediction in Huntington's disease: what are the limitations imposed by pedigree structure? AB - The major factor limiting use of the polymorphic DNA sequence D4S10, genetically linked to the Huntington's disease (HD) locus, in clinical practice is the fragmented nature of HD families. A population survey in South Wales suggested that genetic prediction would only be possible in 15% of adults at risk as a result of this. We have analysed pedigrees from 151 families, containing 482 adults between 18 and 45 years of age who were at high risk of developing HD, 157 of whom had attended genetic counselling clinics. Thirty-seven percent of adults at high risk in these kindreds had the appropriate pedigree structure needed for presymptomatic testing. It should be possible to perform fetal exclusion tests in about 80% of pregnancies at risk. PMID- 2973559 TI - Brain energy metabolism and dopaminergic function in Huntington's disease measured in vivo using positron emission tomography. AB - A 48-year-old man with typical Huntington's disease was investigated with computed tomography (CT) and positron emission tomography. Regional cerebral blood flow, oxygen extraction, oxygen and glucose utilisation, L-Dopa uptake, and dopamine (D2) receptor binding were measured using several positron-labelled tracers. CT showed slight atrophy of the head of caudate but no cortical atrophy, although distinct frontal lobe dysfunction was present on psychometric testing. Oxygen and glucose metabolism and cerebral blood flow were decreased in the striata and to a lesser extent in frontal cortex. Cerebral blood flow was in the low normal range throughout the remainder of the brain. A normal metabolic ratio was found in all regions, since the changes in glucose utilisation paralleled those in oxygen consumption. The capacity of the striatum to store dopamine as assessed by L-[18F]-fluorodopa uptake was normal, but dopamine (D2) receptor binding was decreased when compared to normal subjects. PMID- 2973562 TI - [Transactivation of p'R promoter of phage lambda]. AB - The host-vector system for efficient expression of the cloned genes under the control of transactivated promoter p'R of bacteriophage lambda has been elaborated. The Q protein activating p'R promoter is coded by the defective prophage constructed in vitro by means of excision of the late phage genes between the distant sites of the restriction endonuclease MluI and change of the central SalI fragment carrying the kill gene for the kanamycin resistance gene. The general recombination system is impaired during the change, thus the bacteriophage DNA can be obtained from the induced RecA cells as a plasmid DNA. The induction of the prophage results in a sharp increase of beta-lactamase synthesis (30% of soluble cell protein) under the control of p'R promoter in a plasmid derived of pBR322. PMID- 2973564 TI - The E. coli multitest: a set of strains to characterize diverse genotoxic effects. AB - A set of E. coli strains was developed by Toman et al. (1985) to study the effects of chemical and physical agents on forward mutation, homologous recombination and induction of the SOS system. New tester strains have been constructed to improve this test system in order to explore quantitative genotoxicity spectra. Through the use of these strains: (i) SOS induction can be specifically detected without interference from mutagenesis; (ii) SOS-dependent and SOS-independent mutational events can be distinguished; (iii) the sensitivity of the recombination system has been considerably increased. PMID- 2973563 TI - Anticarcinogenic potential of DNA-repair modulators. AB - Effects of compounds that inhibit repair of DNA lesions in cells have been reported frequently. The consequences include altered incidence of carcinogenicity in vivo, tumorigenic transformation of cultured cells, mutations, and increased lethality as well as sister-chromatid exchanges and chromosome aberrations. This literature is reviewed here, with major emphasis on methylxanthines (caffeine in particular) and nicotinamide analogs. Existing information is also summarized on a novel potent repair inhibitor, beta lapachone. Compounds that inhibit both DNA replication and repair are not discussed in detail since they have been reviewed often, but miscellaneous inhibitors of repair are summarized in a table. The relatively small number of experiments performed on the anticarcinogenic effects of methyl-xanthines and nicotinamide analogs gave very conflicting results. Some investigators report decreased carcinogenicity of DNA-damaging agents when caffeine was provided, but others obtained the opposite effect. The three studies with nicotinamide analogs all reported enhanced tumorigenicity of carcinogens. The data are too few to enable firm conclusions to be drawn regarding the possibility of using repair inhibitors to prevent cancer in humans. Variations of experimental conditions, carcinogens, cells, etc. have provided conflicting results. The possibility of cancer prevention is, nevertheless, so important that further investigations with DNA-repair inhibitors, particularly with human cells, seem very well justified. PMID- 2973565 TI - Interaction at a distance between lambda repressors disrupts gene activation. AB - The lambda repressor is an activator as well as a repressor of transcription. The activation function is blocked by interaction with another lambda repressor molecule bound upstream on the same DNA molecule. This example of negative control at a distance involves formation of a DNA loop. PMID- 2973566 TI - Killing of antigen-reactive B cells by class II-restricted, soluble antigen specific CD8+ cytolytic T lymphocytes. AB - Cytolytic T lymphocytes (CTLs) are generally thought to recognize cellular antigens presented by class I MHC molecules. A number of studies, however, have revealed responses of considerable magnitude involving both CD8+ and CD4+ CTLs with class II restriction, suggesting that class II-restricted CTLs recognizing exogeneous protein antigens may exist. As class II antigens are normally expressed on limited types of cells such as B cells and macrophages, such CTLs might be expected to exert a suppressive effect on antibody responses. Here we report that stimulation of mouse lymphocytes with a soluble antigen induced CD8+ and CD4+ CTLs specific for the antigen with class II restriction. The specific lysis was far more efficient when target B cells specifically recognized the antigen than when they did not, indicating that the primary targets for these CTLs are probably B cells expressing immunoglobulin receptors reactive for the same antigen molecule. These results suggest that the natural occurrence of such CTLs during immune responses may explain antigen-specific suppression on antibody responses by T cells. PMID- 2973567 TI - Laminin receptor on platelets is the integrin VLA-6. AB - Adhesion of platelets to the subendothelial matrix of an injured vessel wall is an essential step in triggering the formation of a haemostatic plug. Fibronectin, collagen and laminin are three major components of the subendothelial matrix which support platelet adhesion. Receptors for fibronectin and collagen have been identified on platelets and are included in the integrin family. Here we report that adhesion of platelets to laminin is inhibited by a rat monoclonal antibody against the integrin family member, VLA-6. This antibody does not affect platelet adhesion to fibrinogen, fibronectin or to type I and III collagen. Binding to laminin does not require platelet activation and is not inhibited by fibronectin and laminin cell-attachment peptides. Platelet adhesion to laminin is supported by Mn2+, Co2+ and Mg2+, but not by Ca2+, Zn2+ and Cu2+. This cation preference is distinct from that characteristic for other platelet-adhesive glycoproteins. PMID- 2973568 TI - Pain syndromes in multiple sclerosis. AB - To determine the prevalence and nature of pain in multiple sclerosis, we evaluated by questionnaire, interview, and chart review 159 patients residing in Middlesex County and followed in the MS Clinic at University Hospital, London, Ontario, Canada. Eighty-eight patients (55%) had either an acute or chronic pain syndrome at some time during their disease. Fifteen patients (9%) with acute pain syndromes had episodes of paroxysmal tic-like pain diagnosed in seven as trigeminal neuralgia. Chronic pain syndromes, present for a mean duration of 4.9 years, occurred in 76 patients (48%) and included dysesthetic extremity pain (29%), back pain (14%), painful leg spasms (13%), and abdominal pain (2%). MS patients with pain were similar to the pain-free group in mean age of onset (34.0 versus 31.9 years), average duration of disease (13.3 versus 12.1 years), spinal cord involvement (97% for each group), and mean rating on Kurtzke Disability Status Scale (4.2 versus 3.5). They differed in sex ratio with a higher female-to male ratio in the pain group (3:1 versus 1.4:1). Chronic pain is a common feature of well-established MS and is usually associated with a myelopathy. Therapy must be individualized for each specific pain syndrome. PMID- 2973570 TI - [Immunologic correlations of Lp(a) lipoprotein and plasminogen]. PMID- 2973569 TI - Hepatitis B virus: the importance of age at infection. AB - Recent studies have demonstrated the importance of age at infection with hepatitis B virus (HBV). Age affects whether the infection is self-limited or results in the chronic carrier state, the severity of the acute infection, and the incidence of various sequelae of the chronic carrier state. In particular, although the acute infection is more severe in adults, infections in infants and preschool children carry much greater risks of chronic carriage which increases the risk of primary hepatocellular carcinoma and cirrhosis later in life. This has two important implications for areas where HBV is endemic. First, more impact can be gained by vaccinating infants and preschool children than by vaccinating healthy adults. Second, if funds are limited, greater impact will be gained by immunising a larger number of children with low doses of vaccine so that they are protected during the early years of life when the risk of chronic carriage is highest, rather than using the standard dose in a smaller number of children even though protection may be longer lasting with standard doses. These two considerations provide the basis for an efficient strategy for control in communities or countries where HBV is endemic or hyperendemic. PMID- 2973571 TI - Mechanisms of bronchial hyperreactivity in cystic fibrosis. AB - We studied 14 patients with cystic fibrosis (CF) who had evidence of bronchial hyperreactivity on a standardized histamine challenge. Patients had a histamine challenge on the first day. Then they were pretreated with either 0.25 mg ipratropium bromide or 0.5 mg fenoterol hydrobromide on 2 separate days, and the histamine challenge was repeated. Baseline forced expiratory volume in 1 sec was similar on the 3 days; however, there was a small but significant (P less than 0.05) improvement after fenoterol. Mean PC20 on the control day was 1.50 mg/ml, which increased significantly after pretreatment with ipratropium (2.88 mg/ml, P less than 0.01) and fenoterol (3.64 mg/ml, P less than 0.005), indicating protection against histamine-induced bronchial hyperreactivity. The six CF patients with "coexistent asthma," as defined by recurrent episodes of wheezing responsive clinically to bronchodilator therapy, had no significant protection from ipratropium, whereas the eight "nonasthmatic" CF patients were protected by both ipratropium and fenoterol. We postulate that at least two mechanisms contribute to histamine-induced bronchial hyperreactivity in patients with CF, one related to vagally mediated reflex bronchoconstriction and another that acts independently of this mechanism. PMID- 2973572 TI - Efficacy of low-dose Aracytine in 3 cases of refractory anemia in transformation in children. AB - A treatment with cytosine arabinoside in low doses is administered to 3 children affected by refractory anemia with myeloblast in transformation. The remission is obtained in 3 cases. One child relapsed at 24 months. The other 2 children are in persistent remission at 40 and 48 months. This relatively non-aggressive mode of treatment seems to have a place in children affected by myelodysplasia and for whom allografting from an HLA-identical donor is not available. PMID- 2973573 TI - Structural bases of a long-stretched deletion: completing the lambda plac5 DNA primary structure. AB - In studying molecular mechanisms of specialised transduction, the lacI (E. coli) Ea47 (lambda) DNA junction in transducing bacteriophage lambda plac 5 has been structurally elucidated, thus yielding the complete sequence of lambda plac 5 DNA including the lac5 substitution, a well-known segment of lambdoid vectors. The lambda plac5 DNA is shown to consist of 19368 bp (lambda left arm) + 3924 bp (lac5 substitution) + 25353 bp (lambda right arm), totally amounting to 48645 bp. The presence of the phage rho bL promoter near to the right end of the lac5 insert is shown. The lacI gene distal end in lambda plac5 proved to be much longer than it was postulated earlier, coding for 224 C-terminal amino acid residues of lac repressor. Both the recombination studied in this paper and the earlier studied abnormal prophage excision (2, 3) occur near to Chi-like structures (chi*lacI and chi*lom, respectively). On the basis of the data obtained, a key role of the E. coli RecBCD system and Chi-like sequences in the formation of deletions in bacterial cells is suggested. PMID- 2973574 TI - Interactions of water-soluble metalloporphyrins with nucleic acids studied by resonance Raman spectroscopy. AB - The resonance Raman spectra of water-soluble porphyrins, M(TMpy-P4) (M = Cu(II), Ni(II) and Co(III] and their mixtures with poly(dG-dC)2, poly(dA-dT)2 and calf thymus and salmon DNAs were measured using a divided rotating cell to determine the magnitudes of frequency shift and intensity variation resulting from M(TMpy P4)-nucleic acid interactions. Bands II(C beta-H bending, approximately 1100 cm 1) and VIII(C beta-C beta stretch, approximately 1570 cm-1) show a large and small upward shift, respectively, when Cu(TMpy-P4) and Ni(TMpy-P4) are intercalated at the G-C sites. In contrast, these bands show a small upward and downward shift, respectively, when Co(TMpy-P4) is groove-bound at the A-T sites of nucleic acids. Both Bands V (approximately 1260 cm-1) and IX (approximately 1646 cm-1) which originate in the N-methylpyridyl group always show small downward shifts due to coulombic interaction between the N-CH3+ group of TMpy-P4 and the PO2 group of the nucleic acid. PMID- 2973576 TI - [Ceftazidime. Evaluation and perspectives]. PMID- 2973575 TI - Screening of pharmacological agents given peripherally with respect to TCDD induced wasting syndrome in Long-Evans rats. AB - A salient sign of fatal 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) intoxication is dramatic body weight loss accompanied by hypophagia. Yet, the nature of this wasting syndrome is unknown. As all of the current leptogenic (weight reducing) drugs exert their action by affecting aminergic neurotransmission, this study set out to screen the reversibility of TCDD-induced anorexia with the following agents modulating aminergic neurotransmission: amphetamine, amperozide, chlordiazepoxide, clonidine, haloperidol, morphine, PCPA, phenoxybenzamine, reserpine and sotalol. In addition, dexamethasone, indomethacin, and insulin were included in the drug battery. The agents were administered subcutaneously to adult male Long-Evans rats over a period lasting from 3 to 14 days. Half of each drug group was concomitantly exposed to a lethal dose of TCDD (20 micrograms/kg). None of the regimens were able to mitigate the wasting syndrome. TCDD proved to markedly diminish the nocturnal feed intake while practically sparing daytime feed consumption. Insulin increased the daytime feeding of TCDD-exposed rats, and the termination of treatment resulted in almost total aphagia in this group. Amphetamine, dexamethasone, PCPA, and reserpine caused weight loss in drug control rats and aggravated the action of TCDD. However, clonidine had no effect on the weight of control rats but accelerated weight decline in TCDD-cotreated animals. TCDD seemed to have a somewhat minor influence on drinking than on feeding. Clonidine stimulated water intake in controls but not in TCDD-exposed rats. These results suggest that aminergic neurotransmission is not specifically or crucially affected by TCDD, but further studies are needed to confirm this conclusion. PMID- 2973577 TI - [Structure-activity relationship of ceftazidime. Consequences on the bacterial spectrum]. AB - The structure-activity relationships of antibiotics are currently known, especially for cephalosporins. Ceftazidime, which is a new semi-synthetic drug, possesses as side chains an aminothiazole ring and a propylcarboxy moiety at position 7, and at position 3 a pyridine group similar to that of cephaloridine: these structural modifications of the cephalosporin nucleus confer upon ceftazidime increased affinity for penicillin-binding-proteins of Gram negative bacilli, a high stability to beta-lactamases and an enlarged spectrum of antibacterial activity including Pseudomonas and Acinetobacter. The presence at position 3 of a pyridine group is responsible for a rapid intrabacterial penetration of ceftazidime and for favourable pharmacokinetic properties. PMID- 2973578 TI - [Plasmid resistance to 3d generation cephalosporins]. AB - Beta-lactamases still play an important part in medical bacteriology, as shown by the emergence, since 1983, of plasmid-mediated beta-lactamases with an enlarged spectrum (SHV-2, CTX-1, etc.). Such enzymes are only produced by enterobacteria and, more specifically, by Klebsiella pneumoniae. This phenomenon, described in Europe and in Africa, is certainly more widespread than it would appear, as some strains are now known to be less sensitive to third generation cephalosporins (MIC 1 to 4 mg/l). Despite differences in behaviour (cefotaximase and ceftazidimase phenotypes), resistance to amino-, carboxy- and ureido-penicillins is associated with reduced sensitivity or resistance to oxyimino beta-lactams (cefotaxime, ceftriaxone, ceftazidime, aztreonam), but cefamycins and imipenem are untouched. Being sensitive to enzyme inhibitors (e.g. clavulanic acid), these beta-lactamases can easily be detected and some infections (notably urinary tract infections) can probably be treated using these inhibitors. These enzymes show modified kinetic constants (better affinity and quicker hydrolysis) against penicillins, third generation aminothiazolimino-cephalosporins and aztreonam. The producing strains are mutants, with aminoacid 1 to 2 substitutions, of those which produce the usual plasmid-borne and transposable beta-lactamases (TEM or SHV). Because these beta-lactamases are plasmid-mediated, enzyme production mechanisms are spreading among enterobacteria species in relation to other resistance markers (tobramycin, netilmicin, amikacin). Strains which produce these new enzymes are mainly isolated from patients treated in intensive care units. PMID- 2973579 TI - [Behavior of ceftazidime in regard to different classes of beta-lactamases. The situation in 1988]. AB - Following Ambler's observations, B-lactamases can be divided in four classes, probably derived from a very small number of ancestral genes. Class A includes the TEM-type beta-lactamases (TEM-1, TEM-2), PIT-2/SHV-1 and others which do not hydrolyse ceftazidime and other third-generation cephalosporins. The new plasmid mediated beta-lactamases markedly active against third-generation cephalosporins also belong to class A and the primary structures of few of them are known. All the class A beta-lactamases are highly susceptible to the action of beta lactamase inhibitors: clavulanic acid, sulbactam, YTR-830, and are devoid of any hydrolytic properties for cephamycins and imipenem. Third-generation cephalosporins are slowly hydrolyzed by the class C beta-lactamases: the inducible cephalosporinases. Thus only organisms which produce derepressed cephalosporinases are resistant to third-generation cephalosporins, but individual variations are known. PMID- 2973580 TI - [Sensitivity of Pseudomonas aeruginosa and Klebsiella spp. to ceftazidime. Current status in France]. AB - Ceftazidime was tested against 2,224 strains of Pseudomonas aeruginosa obtained from 17 hospitals in April, May and June, 1986 and against 607 strains of Klebsiella pneumoniae and 234 strains of K. oxytoca obtained from 16 hospitals in October, 1987. The MIC's of ceftazidime against P. aeruginosa were distributed normally, with an MIC50 of 2 mg/l and an MIC90 of 4 mg/l. Depending on critical concentrations, 80 per cent of strains were sensitive, 11.4 per cent were of intermediate sensitivity and 0.54 per cent were resistant. There were few differences in results between hospitals. Ninety-two per cent of resistant strains and 45 per cent of intermediate strains (as opposed to 6 per cent of all strains) produced a high-level constitutive cephalosporinase with little variations between centres. The MIC's of ceftazidime against K. pneumoniae and K. oxytoca had a bimodal distribution: 91 per cent of strains were sensitive to 0.25 mg/l, 6 per cent of strains showed intermediate sensitivity and 3 per cent were resistant. All intermediate and resistant strains produced a very broad spectrum beta-lactamase which hydrolyzed some of the third generation cephalosporins: K. pneumoniae 36 CTX-1, 5 SHV-2, and 14 strains producing a recently identified beta lactamase "CAZ-5/SHV-4"; K. oxytoca 3 CTX-1. These strains were isolated in 10 of the 16 hospitals which took part in the 1987 study. Comparison of these results with those of studies performed in 1984 and 1985 showed a moderate increase in the number of intermediate sensitivity strains of P. aeruginosa and the occasional occurrence, of the epidemic type, in some hospitals of Klebsiella spp. producing very broad spectrum beta-lactamases which were rare in 1985. PMID- 2973582 TI - [Importance of antibiotic combinations with ceftazidime and usefulness in the prevention of resistance]. AB - Different antibiotics can be used in combinations to extend the antibacterial spectrum, to obtain a synergistic bactericidal effect or to prevent bacterial resistance. Ceftazidime may be combined with other antibiotic to extend its spectrum to Staphylococci and Enterococci, or even anaerobes. A synergistic effect is mainly obtained with aminoglycosides. To prevent the emergence of resistant strains, ceftazidime may be combined with aminoglycosides and the new quinolones. PMID- 2973581 TI - [Theoretical bases for the combination of ceftazidime with other antibiotics. Synergism research]. AB - Combinations of antibiotics have always been difficult to study, and the available methods often give discordant results making interpretation uneasy in the absence of in vitro-in vivo correlations. By studying the bactericidal effects of combinations a better definition of interactions between two antibiotics can be obtained if the concept of dominance is taken into account. Ceftazidime, a time-dependent antibiotic, acts synergistically with aminoglycosides, notably against moderately sensitive strains. This synergistic effect results from acceleration of the early bacterial kill and from blockage of the late regrowths. With quinolones, the synergistic effect does not result from blockage of late regrowths. However, the significance of these results needs to be confirmed by clinical trials. PMID- 2973583 TI - [Ceftazidime on animal experimental models]. AB - Experiments on animal models represent an important stage in the development of antibiotics. As regards ceftazidime, they have confirmed that its activity is time-dependent in vivo, that there is no post-antibiotic effect and that combining this cephalosporin with an aminoglycoside is useful in neutropenic animals, notably in case of Pseudomonas infection. The potential usefulness of ceftazidime in the treatment of meningitis has also been demonstrated. However, animal models have limited value when one tries to extrapolate to man the therapeutic results obtained. In particular, they provide no information on optimal dosage, critical concentrations or desirable duration of treatment. PMID- 2973584 TI - [Pharmacokinetics of ceftazidime in the cerebrospinal fluid]. AB - A compound potentially effective in the treatment of meningitis should reach high concentrations in CSF. In animal experimental models, ceftazidime levels of 5-25 micrograms/ml were demonstrated during administration of doses generating serum levels similar to those achieved in humans. Human studies later confirmed CSF levels of 5-10 micrograms/ml in patients with meningitis. Ceftazidime CSF kinetic properties are adequate for use of this compound in patients with meningitis due to susceptible organisms. PMID- 2973585 TI - [Pharmacokinetics of ceftazidime in healthy and renal failure subjects]. AB - Ceftazidime is poorly bound to plasma proteins, is not metabolized and is excreted mostly by glomerular filtration. Its elimination half-life is about 1.5 2 hours, and their kinetics are modified by renal impairment. The dosage recommended in adult is 1 to 2 g 8-hourly, to be adjusted to the needs of individual cases. The interval between doses must be extended to 12 hours in patients with mild renal impairment and to 36-48 hours in those with severe renal failure. A supplementary dose of ceftazidime is required at the end of each haemodialysis session. In patients under chronic peritoneal dialysis, the loading dose (10 mg/kg) is followed by continuous administration of 5 mg/kg into each dialysis cavity. PMID- 2973586 TI - [Pharmacokinetics of ceftazidime in children and newborn infants. Study in 14 patients and review of the literature]. AB - A pharmacokinetic study of ceftazidime was conducted in 1983, together with a clinical study, on 7 neonates (3 of them premature), 4 infants and 3 children. The antibiotic was administered by slow intravenous injection in mean doses of 30 mg/kg twice in 24 h in neonates and thrice in 24 h in older children. Blood samples were collected by micropuncture, and assays were performed by the microbiological method on agar plates. Curves of plasma concentrations over time showed two slopes compatible with a two-compartment model. The alpha half-life was the same in neonates and infants; the beta half-life varied from 1.9 to 5 h. The highest values were observed in the 3 youngest and most immature neonates. In infants and older children the mean beta half-life was the same as in adults: 1.4 +/- 0.19 h. The results of several studies performed on neonates differed as regards the influence of term, post-natal age and bodyweight on the pharmacokinetic constants of ceftazidime. A dose of 25 to 50 mg/kg twice a day administered to premature and full-term neonates during the 1st week of life gives therapeutically effective concentrations. Doses of 30 to 50 mg/kg 3 or 4 times a day are necessary in infants and older children. Dosage should be adjusted to renal maturity and renal functions as well as to the infection treated. PMID- 2973587 TI - [Pharmacokinetics of ceftazidime in cystic fibrosis]. AB - Ceftazidime has been recognized as an invaluable antibiotic for the treatment of lower respiratory infections in patients with cystic fibrosis. In these patients the apparent volume of distribution of ceftazidime is increased. Its clearance is besides increased, with a significant reduction in the area under concentrations curve. The distribution of ceftazidime in the bronchi seems to be very satisfactory. It results from these data that dosage in children with cystic fibrosis may reach 200 to 300 mg/kg/day divided into 3 or 4 injections. The intrabronchial administration of ceftazidime has not proved effective. PMID- 2973589 TI - [Treatment of nosocomial pneumopathies caused by Pseudomonas aeruginosa with ceftazidime. 50 cases in pulmonary resuscitation]. AB - Fifty patients under prolonged mechanical ventilation who developed nosocomial lung infections caused by Pseudomonas aeruginosa were treated with ceftazidime. This cephalosporin was used alone in 18 cases and combined with another antibiotic (usually an aminoglycoside) in 32 cases. Favourable results were obtained in 40 cases, with regression of the clinical signs and eradication of the organism between the 5th and 10th days of treatment. Eight of the 10 failures occurred in patients with impaired ventilation due to chronic obstructive lung disease. Secondary superinfection was observed in 12 cases. Among the 257 strains of P. aeruginosa isolated in our respiratory intensive care unit over an 18-month period, 88 per cent were sensitive to ceftazidime, and this figure remained stable after the period when ceftazidime was given. These in vitro findings probably account for the remarkable results obtained in infections which up to now had mortality rates of 30 to 60 per cent, depending on the patient's underlaying condition. PMID- 2973588 TI - [Controlled randomized prospective study of a ceftazidime-pefloxacin combination versus a ceftazidime-amikacin combination in the empirical treatment of nosocomial pneumonias and septicemias of resuscitation. Preliminary results]. AB - The preliminary results obtained in 212 patients from 20 intensive care units, suffering from pneumonia or nosocomial septicemia and treated at random with either ceftazidime + pefloxacin (CP) or ceftazidime + amikacin (CA) were analyzed. After exclusion of patients who did not comply with the protocol and of cases where no organism was isolated, 57 assessable patients received CP and 72 received CA. The cure rates achieved in infections due to a single organism were 85 per cent in the CA group and 63 per cent in the CP group. In infections due to multiple organisms, the cure rate was 74 +/- 2 per cent with both antibiotic regimens. PMID- 2973590 TI - [Ceftazidime in the treatment of purulent meningitis]. AB - Third generation cephalosporins have modified the first-line treatment of bacterial meningitis in adults and children. Among these compounds, ceftazidime has a particularly wide antibacterial spectrum, and clinical results have confirmed its superiority, notably in the treatment of meningitis caused by Pseudomonas spp. It is an excellent first-line empirical treatment while awaiting bacteriological results, but in view of the severity of these diseases and of the dangerous pathogens that are often involved, it seems preferable to combine ceftazidime with an aminoglycoside. Route of administration (systemic or in situ) and duration of treatment must be tailored to each individual case. PMID- 2973591 TI - [Ceftazidime in severe infections in burnt patients]. AB - Infections being common and severe in burnt patients, their control is of paramount importance. In 49 burnt patients treated with ceftazidime 60 treatments (43 curative and 17 preventive) were prescribed. Ceftazidime was used as first line therapy, usually combined with an aminoglycoside (amikacin as a rule). The overall success rate was 81 per cent, and only 5 cases (8.6 per cent) could be regarded as true failures. These results, comparable to those found in the literature, are discussed in detail. No significant change occurred in the bacterial population of our hospital unit during the period of the study (more than 3 years). Our experience confirms that ceftazidime should play a predominant role in the treatment of severe infections affecting burnt patients. PMID- 2973593 TI - [Evaluation of ceftazidime treatment in septicemia expert systems]. AB - The septicaemia expert-system (SES) consists of 18 departments of infectious diseases and microbiology in different French university hospitals. In this context, and compared with the totality of cases analyzed, ceftazidime was used by preference in worrying infections, i.e. often older patients, higher frequency of hospital-acquired infections and of multibacterial infections, greater number of pathogens regarded as difficult to eradicate. The results of ceftazidime treatment in these patients were not different from those obtained in the totality of cases. PMID- 2973592 TI - [Role of ceftazidime in severe infections in children. Review of the literature]. AB - Ceftazidime has the same antibacterial spectrum as the other third generation cephalosporins, but it is the most active of all against Pseudomonas aeruginosa. A review of the literature concerning severe infections shows that ceftazidime has been used in children mainly to treat superinfections on cystic fibrosis, infections in immunocompromised subjects and neonatal infections. The results in large series of patients were highly satisfactory and the drug was well tolerated. Other diseases treated in shorter series were: urinary tract infections in paediatric urology, ENT infections caused by P. aeruginosa, cellulitis and post-operative infections. A multicentre trial involving 344 children has recently been reported, showing very good results (clinical cure in 95 per cent of the cases). Ceftazidime dosage varies from one study to another, but a mean daily dose of 150 mg/kg is often necessary in septicaemias and infections associated with cystic fibrosis. PMID- 2973594 TI - [Treatment of infection in granulopenic patients. Lessons from international studies]. AB - In febrile neutropenic patients the initial antibiotic therapy must be active against several bacteria, notably enterobacteria, Pseudomonas, Staphylococci and Streptococci. Broad-spectrum beta-lactam antibiotics (ceftazidime, ureidopenicillins, ticarcillin-clavulanic acid, imipenem, etc.) have given excellent therapeutic results. Bacteraemias due to Gram-positive cocci are increasing, most of them being related to infection of indwelling intravascular devices. In several studies vancomycin has been systematically prescribed as an empirical treatment of neutropenic patients. New antibiotics, such as fluoroquinolones or teicoplanin have been used in combination with beta-lactams. Non-absorbable antibacterial or antifungal agents, or even diffusible antimicrobial agents, could be used prophylactically in deep induced neutropenia and in the initial phase of bone marrow transplantation. Prospective randomized studies of different regimens are currently in progress to try and determine the best therapeutic and prophylactic measures against infections in neutropenic patients. PMID- 2973595 TI - [First-line treatment of febrile episodes in leukemia in adults. Randomized, multicenter study of ceftazidime in single antibiotic therapy versus a cefotaxime amikacin combination]. AB - A prospective study was conducted in 10 haematology departments of university hospitals on 174 leukaemic patients with prolonged bone marrow aplasia and presenting with a febrile episode. The patients were allocated at random to either ceftazidime or the cefotaxime-amikacin combination. The two treatment group were similar as regards age, sex, underlying blood disease, duration of neutropenia, presence of a venous catheter, type of digestive tract contamination, clinical and bacteriological findings. Results were assessed mainly on the course of the fever at 48 hours and on the clinical and bacteriological changes observed until the patients came out of aplasia. Documented infections were specifically analyzed. There was no significant difference in terms of success or failure between the two treatment groups. Ceftazidime administered as monotherapy proved as effective as the cefotaxime amikacin combination in the empirical first-line treatment of febrile episodes in leukaemic patients with neutropenia. PMID- 2973596 TI - [Randomized prospective study of ceftazidime versus a cefotaxime-tobramycin combination in acute leukemia in therapeutic aplasia]. AB - In a prospective study, 157 patients with prolonged aplasia (PMN less than 500/mm3 during more than 21 days), hospitalized in a protected environment unit, were randomly assigned to receive ceftazidime alone or cefotaxime + tobramycin for initial febrile episodes. Age, sex, underlying diseases, duration of neutropenia, digestive decontamination regimen, clinical and microbiological characteristics of infections were similar in the two groups. Patients were evaluated for their initial response to antibiotics (defervescence in 48 hours, maintained 7 days) and long term response (prevention of another infection during aplasia). The overall initial response to ceftazidime was 48/71 (68 per cent) and to cefotaxime + tobramycin 55/86 (64 per cent). The long term response to ceftazidime was 33/71 (46.5 per cent) and to cefotaxime + tobramycin 31/86 (36 per cent). In conclusion, ceftazidime alone was as effective as cefotaxime + tobramycin in the first line treatment of febrile episodes in neutropenic patients. PMID- 2973597 TI - [Prospective and comparative study of 2 antibiotic therapy protocols in 66 febrile neutropenic patients. Ceftazidime-vancomycin versus ticarcillin vancomycin-amikacin]. AB - Out of 66 febrile neutropenic patients, 33 were treated with a ceftazidime vancomycin combination (group A) and 33 with a ticarcillin-vancomycin-amikacin combination (group B). There was no significant difference in satisfactory results between the two groups (group A 79 per cent, group B 88 per cent), and both regimens were equally active in all febrile episodes. Reversible side effects (renal and cutaneous toxicity) were observed in 15 per cent of the cases. Two cases of superinfection and one case of resistance occurring during treatment were noted in group B patients. At a time when Gram-positive infections are increasingly frequent among febrile neutropenic patients, the ceftazidime vancomycin combination seems to be as effective as the ticarcillin-vancomycin amikacin triple drug combination. PMID- 2973598 TI - [Antibiotic therapy protocol using ceftazidime 3g/day alone or in combination with vancomycin or amikacin. In febrile episodes in neutropenic patients]. AB - In a preliminary study of 21 febrile episodes in neutropenic patients ceftazidime used as empirical treatment in doses of 3 grams per day succeeded in controlling fever in 74 per cent of the cases. Laboratory studies performed in patients with Gram-negative septicaemia showed clinically effective plasma concentrations of the antibiotic. A trial of ceftazidime (3 g/day) administered alone or combined with amikacin or vancomycin is currently in progress in two medical centres. No statistically significant conclusions could be reached from an intermediate study. PMID- 2973599 TI - [Ceftazidime after bone marrow graft. Current aspects]. AB - The aplasia which follows bone marrow transplantation is extremely deep and characterized by disruption of all cutaneous and mucosal barriers and by major immunodepression. It is complicated in 30 to 40 per cent of the cases by septicaemia usually caused by Gram-positive organisms. Because the natural defence mechanisms have been suppressed an empirical broad-spectrum antibiotic therapy is usually instituted before microbiological results are available, and this has proved effective in reducing the immediate mortality due to infection. However, nephrotoxic drugs such as cyclosporin A and amphotericin B are also used in this context, and clinicians are looking for antibiotics that have minimal noxious effect on the kidneys. Ceftazidime administered alone or combined with another antibiotic is currently being investigated from this point of view in several clinical trials. PMID- 2973600 TI - [Ceftazidime for the treatment of infections in neutropenic children]. AB - Ceftazidime in doses of 100 mg/kg/day was used, combined with netilmicin 6 mg/kg/day, as first-line treatment in two successive studies conducted on febrile neutropenic children (neutrophils less than 500/mm3). Study n. 1, performed at the Infantile Haematology unit of Saint Louis hospital, Paris, included 75 children. Study n. 2 was a multicentre study involving 88 children from 11 medical centres. The children's age in both studies ranged from 2 months to 16 1/2 years (mean 7 years). The percentage of bacteriologically documented febrile episodes was 45 per cent (34/75 and 39/88), and the most frequent infections were those caused by Gram-positive cocci (56 and 58 per cent respectively of the cases). Vancomycin 40 mg/kg/day was introduced if fever was still present 48 hours after the beginning of the antibiotic therapy. Effective treatments were continued until the neutropenia was corrected. These children were being treated for acute leukaemia, lymphoma, solid tumours or bone marrow aplasia. In study n. 1 apyrexia was obtained in 85 per cent of the cases with the ceftazidime netilmicin combination and in 91 per cent of the cases after addition of vancomycin. The initial therapy was effective in all patients with a documented infection. There were tow super-infections with septicaemia: one due to Streptococcus D, the other to Staph. epidermidis. In study n. 2 73 per cent of the patients were apyretic after the first combination and 85 per cent after vancomycin was introduced. In proven infections the ceftazidime-netilmicin combination was effective in 30 cases and in another 6 cases after addition of vancomycin. Three patients remained febrile until they came out of aplasia. In all cases the bacterial cultures were sterilized by the ceftazidime-netilmicin combination. There was no superinfection. The mean duration of antibiotic therapy was 21 days in study n. 1 and 14 days in study n. 2. The drugs were perfectly tolerated both clinically and biochemically. No death occurred in the two studies. Thus, owing to its broad spectrum, effectiveness and safety ceftazidime is a very useful antibiotic when combined with netilmicin as first-line treatment of febrile neutropenic children. PMID- 2973601 TI - [Empirical antibiotic therapy and ceftazidime]. AB - Ceftazidime is indicated mainly for the empirical treatment of severe infections treated in hospital and of hospital-acquired infections. It can be used alone in numerous severe infections and it offers a good alternative to aminoglycosides and antibiotic combinations. However, there are frequent clinical situations where this drug must be used in combination with vancomycin to cover Staphylococci and Enterococci, and/or with an imidazole derivative to cover anaerobic species. Similarly, when the pathogens are known to be multiresistant the ceftazidime-aminoglycoside combination has a synergistic bactericidal activity and prevents the emergence of resistant strains. This combination is also recommended in cases where the site of infection is difficult to reach or in immunocompromised patients. PMID- 2973602 TI - [Ceftazidime and probabilistic treatment of severe infections in adults]. AB - Probability antibiotic treatments are based on two "best guesses": the nature of the responsible organism and its resistance phenotype. Such treatments are justified in immunocompetent patients with severe infection. The microbiological and pharmacokinetic properties of third generation cephalosporins, and notably ceftazidime, are such that their use as single or combined therapy can be determined. A very wide spectrum and a tendency towards a stronger bactericidal effect are in favour of monotherapy preceded by a short course of bitherapy, especially in cases of hospital-acquired infections. PMID- 2973603 TI - [Probabilistic treatment with ceftazidime of severe infections in children]. AB - Ceftazidime should only be given as "best guess" treatment in infections suspected of being caused by multiresistant bacteria, and notably Pseudomonas aeruginosa. During the last few years, the use of this antibiotic against infections associated with therapeutic bone marrow aplasia or with cystic fibrosis has been thoroughly investigated. As regards neonatal infections, late (hospital-acquired) infections seem to be the best indication. Other major reasons for prescribing ceftazidime are septicaemias in paediatric intensive care units and in surgery, especially when it is feared that P. aeruginosa might emerge. Urinary infection after surgery for complex urinary tract malformations, meningeal infection in neurosurgery, severe otitis media, suspected mastoiditis and otitis of the newborn are special conditions where ceftazidime can be used as first choice therapy. Ceftazidime may be administered alone in certain circumstances, but it is usually given in combination with another antibiotic. In these severe infections, dosage must be high enough to reach the elevated MIC's required by some strains of Pseudomonas. PMID- 2973604 TI - [Probabilistic treatment with ceftazidime of infections in neutropenic patients]. AB - Infection is the most common cause of mortality in neutropenic patients. Although fever does not necessarily mean infection, it must be regarded as its first sign and treated, within hours of its onset, on the basis of probability before a pathogen is isolated. The first-line treatment must cover a wide antibacterial spectrum corresponding to the usual bacteriological flora and to the patient's underlying pathology. The risk of Gram-negative septicaemia in infants and elderly people and the frequency of staphylococcal infections in patients with an indwelling central catheter are well-known. The "best guess" treatment should consist of a third generation cephalosporin, notably ceftazidime, and an antistaphylococcal antibiotic. This treatment should be pursued throughout the period of neutropenia. Due to advances in antibacterial therapy, more aggressive chemotherapeutic regimens can now be prescribed to improve the prognosis of acute blood diseases and of numerous carcinomas. PMID- 2973605 TI - [Cellular immunity indices and their information value in autoimmune diseases of the thyroid (a lecture)]. PMID- 2973606 TI - Additional low-abundance human small nuclear ribonucleoproteins: U11, U12, etc. AB - Two-dimensional gel fractionation has revealed the existence of a number (greater than or equal to 8) of additional species of HeLa cell small RNAs that have 5' trimethylguanosine cap structures and are bound by proteins containing Sm epitopes. Therefore, these low-abundance (10(3)-10(4) per cell) RNAs belong to the Sm class of small nuclear ribonucleoproteins (snRNPs), whose best-known members are the four highly abundant (approximately 10(6) per cell) particles required for pre-mRNA splicing. The complexity of Sm snRNPs in mammalian cells is thus not greatly different from that previously established for lower eukaryotes. Two of the new RNAs, designated U11 (131 nucleotides) and U12 (150 nucleotides), have been sequenced. The U11 and U12 snRNPs have been characterized further by examining their nuclease sensitivity and their possible interactions with other snRNPs. Potential roles for the low-abundance snRNPs in aspects of pre-mRNA processing are discussed. PMID- 2973607 TI - Fluorescence in situ hybridization with human chromosome-specific libraries: detection of trisomy 21 and translocations of chromosome 4. AB - Chromosomes can be specifically stained in metaphase spreads and interphase nuclei by in situ hybridization with entire chromosome-specific DNA libraries. Unlabeled human genomic DNA is used to inhibit the hybridization of sequences in the library that bind to multiple chromosomes. The target chromosome can be made at least 20 times brighter per unit length than the others. Trisomy 21 and translocations involving chromosome 4 can be detected in metaphase spreads and interphase nuclei by using this technique. PMID- 2973608 TI - Release of IgD-binding factor by T cells under the influence of interleukin 2, interleukin 4, or cross-linked IgD. AB - Helper T cells with receptors specific for IgD have immunoaugmenting properties. We have now detected soluble IgD-binding factor in cell supernatants immobilized on nitrocellulose paper by their ability to bind 125I-labeled IgD. IgD-binding factor is released by normal splenic T cells stimulated with recombinant interleukin 2, recombinant interleukin 4, or crosslinked IgD in amounts paralleling the induction of IgD receptors on the cells. IgD receptors are constitutively produced by antigen-specific helper T-cell hybridomas 2H10 and A3.4C6. Incubation of these hybridoma cells with recombinant interleukin 2 increases release of IgD-binding factor while reducing expression of IgD receptors. Specificity of the binding factor for IgD is established by (i) competitive inhibition; (ii) the ability of the binding factor to bind radiolabeled IgD and not monoclonal IgE, IgG2a, or polyclonal IgG; and (iii) the removal of the binding factor on passage through an IgD-Sepharose column and recovery in a subsequent acid eluate. PMID- 2973610 TI - Underfeeding and exposure to short photoperiod alters rat pineal and Harderian gland lysosomal enzyme activities. AB - Harderian gland (HG) weight and lysosomal enzyme activity were evaluated after 21 day-old female rats were singly caged in a long (LP; 14:10 LD) or short (SP; 8:16 LD) photoperiod and fed on one of two dietary regimens (fed ad libitum or 50% underfed) for 50 days; an additional fed and an underfed group of animals in LP were injected every afternoon with 100 micrograms melatonin. Absolute HG weights were significantly lower in all underfed groups compared to their respective fed controls or to the LP fed control group. Absolute HG weights of underfed rats in SP were significantly lower than the underfed rats in LP. Relative HG weights (mg/100 g body wt) were significantly higher in the underfed saline or melatonin treated groups compared to their respective fed controls; however, HG of the underfed SP group were not different from SP-fed controls. No significant differences in HG acid phosphatase, hexosaminidase, and beta-glucuronidase activities were observed in any of the treatment groups maintained in LP. Acid phosphatase, hexosaminidase, and beta-glucuronidase activities were significantly elevated in HG of underfed animals maintained in SP compared to their respective fed controls or to the LP-underfed group. Both the underfed control and the underfed-melatonin treated groups had lower pineal protein values than their respective fed groups; underfed animals in 8:16 LD had similar pineal protein values compared to those of the fed control group in SP. Significant effects of photoperiod and underfeeding with no interaction between these variables were observed on pineal acid phosphatase. The fed group maintained in 8:16 LD had significantly higher acid phosphatase activity than the fed group kept in 14:10 LD. In conclusion, underfeeding resulted in severely reduced body weights and absolute Harderian gland weights. Increased activity in certain lysosomal enzymes occurred in both the pineal and Harderian gland and in some instances this was dependent upon the light cycle and dietary regimen to which the animals were exposed. PMID- 2973609 TI - Expression of CD3-associated antigen-binding receptors on suppressor T cells. AB - Three suppressor T (Ts)-cell hybridomas specific for 4-hydroxy-3-nitrophenyl acetyl (NP) hapten were selected for surface expression of cluster determinant 3 (CD3) by using antibody (anti-CD3) or antigen (NP-bovine serum albumin) panning procedures followed by cloning at limiting dilution. The CD3-selected Ts hybridomas showed a 1-2 logarithmic enrichment in suppressor activity when compared to the parent lines; they also specifically bound NP-coupled sheep red blood cells in rosette assays. This antigen-binding ability could be down modulated by anti-CD3 antibody. Similarly, surface expression of CD3 was specifically down-modulated by preincubation of these hybridomas with antigen. Anti-CD3 monoclonal antibody under reducing conditions coprecipitated a broad band of 38-50 kDa associated with two CD3 (25 and 16 kDa) bands. T-cell receptor, anti-alpha-specific monoclonal antibody also immunoprecipitated a broad band in the 41 to 49-kDa region. The combined results suggest that, like helper and cytotoxic T lymphocytes, Ts cells also bear antigen-specific receptors associated with CD3 molecules. PMID- 2973611 TI - Effects of two nonsulfhydryl angiotensin-converting enzyme inhibitors, CGS 14831 and CGS 16617, on myocardial damage and left-ventricular hypertrophy following coronary artery occlusion in the rat. AB - The present study was designed to examine the effects of two new angiotensin converting enzyme (ACE) inhibitors, CGS 14831 and CGS 16617 (3 mg/kg i. v. 1 min prior to occlusion and 4 and 24 h after occlusion), on myocardial ischemic (MI) damage and left-ventricular hypertrophy in rats. Administration of CGS 14831 or CGS 16617 inhibited angio-tensin-I-induced pressor responses by 40-100% for 4 h after each dose. Myocardial creatine phosphokinase (CK) levels were 10.6 +/- 0.6 U/mg protein in sham-MI animals, and following coronary artery occlusion for 48 h were decreased to 4.1 +/- 0.2 U/mg protein in MI + vehicle animals (p less than 0.01). CGS 14831 and CGS 16617 attenuated the decrease in CK content and resulted in 47 and 40% sparing, respectively, of the left-ventricular free wall. Neither agent attenuated the left-ventricular hypertrophy which developed following coronary artery occlusion. These data indicate that the nonsulfhydryl ACE inhibitors CGS 14831 and CGS 16617 have a significant cardioprotective effect in rats surviving 48 h, and suggest a potential therapeutic usefulness of these agents for the treatment of ischemia-induced heart failure. PMID- 2973612 TI - Comparative study finds McKenzie protocol 97% effective against lumbar syndrome. PMID- 2973614 TI - Does vanadyl affect adenylate cyclase? AB - While the stimulatory effect of vanadate, an anion of pentavalent vanadium, on adenylate cyclase (AC) has been repeatedly demonstrated in various tissues only a few studies have been hitherto devoted to the effect of vanadyl, a cation of tetravalent vanadium, but these have provided contradictory results. In the present experiments synaptic plasma membranes from normal rat cerebral cortex were used for estimation of the vanadyl effect (in the concentration range from 10(-5) mol.1(-1) to 10(-3) mol.1(-1) on the basal adenylate cyclase activity. Four types of incubation media were used. In the presence of Tris-maleate and creatine phosphate + creatine phosphokinase (CP + CK) maximal stimulation (33%) was reached at 10(-4) mol.1(-1). In the same buffer but in absence or (CP + CK) maximum was already obtained at 10(-5) mol.1(-1) (49%); at 10(-3) mol.1(-1) no effect was observed. In Tric.HCl buffer with (CP + CK) maximal stimulation appeared at 10(-5) mol.1(-1), whereas at 10(-3) mol.1(-1) inhibition (-25%) was observed. In a medium containing Tris.HCl without (CP + CK) the biphasic nature of vanadyl effect was less markedly expressed: maximal stimulation (+55%) occurred at 10(-4) mol.1(-1). Thus vanadyl stimulates AC, but at relatively low concentrations (10(-5)-10(-4); at higher concentration it tends to exert an inhibitory action. Vanadate had a qualitatively similar effect, but the stimulation was more pronounced and the tendency to inhibition was shifted to higher concentrations. PMID- 2973613 TI - Specific effect of putative 5-HT1A agonists, 8-OH-DPAT and gepirone, to increase hypertonic saline consumption in the rat: evidence against a general hyperdipsic action. AB - Previous reports indicate that 5-HT1A agonists, in addition to benzodiazepines, increase the consumption of hypertonic saline in rehydrating rats. Experiment 1 investigated the effects of 8-OH-DPAT (10-100 micrograms/kg) and gepirone (0.1 3.0 mg/kg) on consumption of water and of saline over a range of concentrations (0.45%-2.8%) in a 30 min drinking test. The two 5-HT1A agonists dose-dependently increased ingestion of two hypertonic salt solutions, but produced little or no increase in the drinking of water, hypotonic or isotonic saline. Experiment 2 demonstrated that 8-OH-DPAT and gepirone did not enhance water consumption in animals given a water preload, or markedly increase drinking quinine-adulterated water. Taken together, the results indicate a selective dose-related effect of the two drugs to increase hypertonic saline drinking; they did not have a general hyperdipsic effect across all salt and water conditions, and they did not increase intake simply because of a low baseline level of consumption. Hence, 5 HT1A agonist act much more selectively than benzodiazepines in their effects on drinking responses. PMID- 2973615 TI - Radioimmunoassay of atrial natriuretic peptide in human plasma: application to studies of volume and blood pressure homeostasis. AB - Sensitive radioimmunoassay for determination of immunoreactive atrial natriuretic peptide (ANP) in human plasma was developed and employed for the study of plasma ANP concentrations in healthy controls under basal conditions (2.4 +/- 0.1 pmol/l) and during volume expansion by saline infusion (9.6 +/- 2.0 pmol/l and 14.2 +/- 1.8 pmol/l, respectively). Plasma renin activity and plasma aldosterone concentration exhibited opposite changes during saline infusion. In pathological states associated with extracellular fluid volume (ECFV) expansion, ANP concentration were significantly higher than in the controls (liver cirrhosis 8.6 +/- 0.9; congestive heart failure 33.1 +/- 4.8; chronic renal failure before haemodialysis 72.2 +/- 6.4 pmol/l). Further volume expansion in liver cirrhosis by saline infusion led to the further increase in ANP (13.3 +/- 1.3 and 16.1 +/- 1.5 pmol/l, respectively) and ECFV reduction by ultrafiltration during haemodialysis in chronic renal failure diminished but did not normalize plasma ANP (22.5 +/- 2.9 pmol/l). In patients with arterial hypertension the concentration of ANP exceeded the normal range by 62.5% and reached 8.0 +/- 0.5 pmol/l on the average. Our results support the suggestion that ANP is an important regulatory humoral mechanism participating in the regulation of sodium, volume and blood pressure homeostasis. PMID- 2973616 TI - Changes in the development of spontaneous motility in chick embryos after the chronic administration of GABA. AB - From the 4th to the 16th day of incubation, GABA was administered continuously to chick embryos in a mean dose of 9.04 +/- 0.98 mg/kg e.w./24 h. On the 17th day of incubation, spontaneous motility was evaluated from the frequency of spontaneous movements as resting motility and motility after the acute administration of GABA (100 mg/kg e.w.), bicuculline (1 mg/kg e.w.) and oxazepam (10 mg/kg e.w.). 1) The chronic administration of GABA reduced the spontaneous motor activity of the experimental embryos to 38.4-47.8% of the control value. To obtain this effect it was sufficient to administer GABA between the 4th and the 8th day of incubation. 2) The inhibitory effect of the acute administration of GABA in the experimental embryos was only half its effect in the controls. Conversely, the relative size of bicuculline activation of motility was distinctly greater in the experimental embryos, which were also significantly more sensitive to oxazepam. The results show that GABA has a dual effect during embryogenesis--a) an early effect between the 4th and 8th day of incubation causing a chronic debt in the development of spontaneous motor activity, and b) an inhibitory effect as a central transmitter, which begins to be manifested in embryonic spontaneous motility of chick embryos on about the 15th day of incubation. PMID- 2973618 TI - Changes in metabolism, food and water intake of C3H mice following transplantation of Gardner lymphosarcoma (LSG). AB - Mice bearing progressing Gardner lymphosarcoma (LSG) increasingly gained in weight in the course of LSG development but their food and water consumption was reduced in comparison with control animals. During the neoplastic growth, oxygen consumption was decreased and the proportion of the metabolically active part (i.e. dried matter) declined. The body weight of tumour-bearing mice successfully cured with methotrexate (MTX) (30 mg/kg body weight, administered on days 1 and 3) was substantially lower than those of non-treated tumourous animals and was similar to that of intact controls. The food and water consumption of mice treated with MTX and that of control animals was similar. Values of the metabolic rate observed in animals treated with MTX were comparable to those recorded in the control group. PMID- 2973617 TI - Effect of chronic ethanol treatment and subsequent ischaemia on phospholipids and cholesterol in the rabbit spinal cord. AB - Rabbits received ethanol p.o. (0.96 g. ml-1, 2.88 g.kg-1) for 30 days. Ischaemia was induced by abdominal aorta ligation for 40 min in animals with or without ethanol treatment. The content of total (TPL) and individual phospholipids, i.e. ethanolamine (PE), choline (PC), serine (PS), phospholipids and sphingomyelin (SM), as well as unesterified cholesterol (UC) was determined in the gracilis fascicle (Fg), and the dorsal (Dp) and ventral (Vp) part of the lumbar and cervical spinal cord. Chronic ethanol treatment resulted in a statistically significant decrease in the PE content in Dp of cervical spinal cord. Cholesterol content was increased in all parts of the spinal cord studied (increased UC/TPL molar ratio). Ischaemia of the spinal cord induced a significant decrease in PI. In ethanolic animals ischaemia decreased the PS content in Dp and Vp of ischaemized lumbar spinal cord. The combined effect of ischaemia and chronic ethanol did not result in a cumulative pattern of changes suggesting a partially opposite influence of both stimuli on lipid metabolism as well as its altered regulation after chronic ethanol treatment in the spinal cord. PMID- 2973619 TI - Glycogenesis and lipogenesis from 14C-glucose in vivo in rats irradiated with fractionated doses of gamma rays. AB - Irradiation with fractionated doses is a specific form of stress and the data concerning these problems are topical for recent radiobiology, radiology and oncology. Interest in this present paper is focused on tissue glycogenesis and lipogenesis from U-14C-glucose in vivo in rats irradiated with fractionated doses of 2.39 Gy once a week. Analyses were done after 1-6 fractions, up to total accumulated doses of 2.39, 4.78, 7.17, 9.76, 11.95 and 14.34 Gy, which means LD50/30 for this experimental model. Fractionated irradiation of rats led to glycogen deposition and increased incorporation of 14C-glucose into the liver, heart and skeletal muscles, but not into brain glycogen. The ascertained changes were not dose-dependent. 14C-glucose was incorporated into the liver and adipose tissue lipids to a small extent, and synthesis of liver cholesterol increased only after the 5th and 6th fractions. A decreased concentration of hepatic lipids, especially of cholesterol, was observed from the 3rd to the 6th fractions. PMID- 2973620 TI - Metyrapone block of thyroid-induced cardiomegaly. AB - Cardiac wet and dry weight, adrenal weights, blood pressure and food consumption were measured in control male rats, rats fed 0.2% dried thyroid powder in the food, rats fed metyrapone (an inhibitor of adrenocortical 11-hydroxylase, Metopirone CIBA, 50 mg/rat per day) and both metyparone + dried thyroid. Metyparone markedly inhibited thyroid-induced cardiomegaly as well as the increase in adrenal weight. Myocardial dry weight and blood pressure differences between the various groups were not significant. The results supply further evidence of the existence of a relationship between the adrenal cortex and thyroid-induced cardiomegaly. Metyrapone also completely inhibited the post thyroid drop in body weight. PMID- 2973621 TI - Hormonal regulation of postnatal development of renal tubular transport processes. AB - Renal tubular transport of p-aminohippurate (PAH) is immature at birth. Repeated saturation of transport sites by treatment with various organic anions is without any influence on the postnatal development of kidney transport capacity. Hormonal regulation of postnatal maturation of PAH transport must therefore be taken into consideration. It was tried to stimulate immature PAH transport by treating rats of different ages with thyroid hormones, corticosteroids or testosterone, respectively. In rats with immature kidney function, renal PAH excretion can be stimulated by daily treatment with thyroid hormones. Experiments on renal cortical slices have shown that PAH excretion is preferentially stimulated by an increase of transport capacity. Whereas thyroid hormones stimulate the renal excretion of PAH both in young and in adult rats, dexamethasone treatment is more effective in rats with immature kidney function. Dexamethasone treatment is without any influence on PAH accumulation in renal cortical slices. Kidney weight and the protein content of kidney tissue was increased after dexamethasone treatment. Repeated testosterone administration did not stimulate the PAH transport in rats of different ages. The data have demonstrated the influence of thyroid hormones or of dexamethasone on renal tubular transport processes in rats with immature kidney function. Treatment with such hormones could be useful in the management of renal insufficiency in full-term and pre-term neonates with immature kidney function. PMID- 2973623 TI - Proceedings of the Czechoslovak Physiological Society. (Kosice, September 9-11, 1987). PMID- 2973622 TI - Effect of isoprenaline on 85Sr accumulation in the myocardium of the adult rat. AB - The administration of a single dose of isoprenaline to adult male rats stimulated 85Sr uptake by their blood, liver and myocardium. Whereas the 85Sr concentration in the blood and liver rose by only 36% and 42% respectively, the increase in the various compartments of the heart was 5-7 times this value. This finding is also documented by the increase in the ratio of the 85Sr concentration in the heart and blood. The increase observed in 85Sr accumulation in the myocardium is comparable to the repeatedly described changes which occur in radioactive calcium after the administration of isoprenaline. PMID- 2973624 TI - [The effect of ionizing radiation on the blood plasma and erythrocytes and on the wall of the abdominal aorta of rats]. AB - On days 7 and 15 after gamma-irradiation (4 Gy) changes were noted in the temperature dependence of the erythrocyte suspension viscosity coefficient, in the electrolyte composition of the abdominal aorta, plasma, and erythrocytes, in Na, K- and Mg-ATPase activity, and in the intensity of fluorescence of 1.8 ANS of erythrocyte ghost of albino rats. The changes were a function of the stage of radiation sickness and were more pronounced on the 15th day following irradiation. PMID- 2973625 TI - [Positive diagnosis and treatment of ocular toxoplasmosis]. PMID- 2973627 TI - [Ocular hypotony]. PMID- 2973626 TI - [Teratogenic factors in ophthalmology]. PMID- 2973628 TI - [Uveitis today]. PMID- 2973629 TI - [Retinal arterial obliterations]. PMID- 2973630 TI - [Computerized diagnosis in retinal and choroidal dystrophies]. PMID- 2973631 TI - [Relation of intraocular pressure, diabetes mellitus and diabetic retinopathy]. PMID- 2973632 TI - [Injuries to the lacrimal ducts. Clinical and therapeutic aspects]. PMID- 2973633 TI - [The therapeutic procedure in scleral wounds]. PMID- 2973634 TI - [Localization and extraction of metallic paraocular foreign bodies]. PMID- 2973635 TI - [Pathologic cataract in pseudosyringomyelic ulceromutilating acropathy]. PMID- 2973637 TI - [Malignant uveal melanoma and cutaneous pigmented nevi]. PMID- 2973636 TI - [Conjunctival lymphosarcoma]. PMID- 2973638 TI - [Acute post-hemorrhagic ischemic optic neuropathy with bilateral blindness]. PMID- 2973639 TI - [Bilateral retinoblastoma]. PMID- 2973640 TI - [Tapioca melanoma of the iris]. PMID- 2973642 TI - Parenteral vaccination of calves against the cattle lungworm Dictyocaulus viviparus. AB - Protection against challenge with Dictyocaulus viviparus larvae was studied in two groups of calves. The first group was vaccinated orally with an irradiation attenuated larval vaccine on two occasions, 28 days apart, as recommended by the manufacturer. Each dose contained 1000 larvae. The second group was vaccinated by two subcutaneous injections of vaccine, also 28 days apart. Compared with unvaccinated calves the protection in both groups of vaccinated calves was similar, ie, over 95 per cent reduction in adult worm burdens after an oral challenge of 3000 to 4000 larvae. These results indicate that the passage of irradiated larvae through the intestine and mesenteric lymph nodes is not necessary for the stimulation of a high degree of immunity and opens up the possibility of parenteral vaccination against this and related diseases. PMID- 2973641 TI - Indicators of inflammation to evaluate the recovery from acute bovine mastitis. AB - The recovery of 39 mastitic cows was evaluated using alpha 1-antitrypsin, N acetyl-beta-D-glucosaminidase (NAGase), plasmin and somatic cell counting for monitoring the inflammatory process. Bacteriological investigation was undertaken during the acute phase and at days 21 and 50 after treatment. In the quarters with a bacteriological cure the concentrations of all inflammatory markers in the milk returned to normal within three weeks. The quarters which harboured subclinical infection at days 21 and 50 after the onset of the disease showed higher values of the inflammatory parameters than uninfected, healed quarters. NAGase was the best predictor of the bacteriological status during the healing period (three weeks after therapy). The study suggests that NAGase determination in each quarter (with interquarter evaluation) possibly in conjunction with a bacteriological investigation three weeks after therapy, would be a good combination for assessing the outcome of treatment. PMID- 2973643 TI - Energy metabolism in disorders of the nervous system. AB - "Energy metabolism" is deranged in a wide variety of disorders of the nervous system. This term refers rather loosely to the pathways responsible for the utilization of the major substrates of brain. Primary disorders of energy metabolism are those in which the primary insult affects the cellular machinery required for energy metabolism. A typical example would be a defect in a gene coding for a mitochondrial protein. Biochemically, defects which appear to be hereditary and which lead to disease of the central nervous system have been described in each of the pathways of energy metabolism: glycogenolysis (the break down of glycogen to glucose); glycolysis (the break down of glucose to pyruvate and lactate); the pyruvate dehydrogenase complex (which oxidizes pyruvate to enter the Krebs tricarboxylic acid cycle); the tricarboxylic acid cycle itself (which completes the oxidation of carbohydrates and other substrates to carbon dioxide); electron transport (which carries out their oxidation to water); the pentose phosphate pathway (an alternate pathway for glucose oxidation); and several "minor" mitochondrial pathways. Clinically, the spectrum of syndromes associated with primary disorders of energy metabolism is wide. Common manifestations include psychomotor retardation, with associated lactic acidosis and/or hypoglycemia. The laboratory abnormalities may be intermittent. Syndromes which have been culled out include congenital lactic acidosis, Leigh disease, intermittent ataxia, Kearns-Sayre-Shy syndrome (KSS), myoclonus epilepsy with ragged red fibers (MERRF), and mitochondrial myopathy-encephalopathy-lactic acidosis-stroke (MELAS). As with other families of inborn errors, both clinical and biochemical heterogeneity occur. Patients with apparently similar clinical syndromes can turn out to have different inborn errors, and patients with abnormalities of the same gene product can have clinically distinguishable syndromes. Secondary disorders are those in which the derangements of energy metabolism are presumably secondary to some other insult but may still be important for the cellular pathophysiology. These include the metabolic encephalopathies and probably a number of well-known neurodegenerative disorders. In the hereditary ataxias, abnormalities of mitochondrial markers are common but do not correlate consistently with the disorders as conventionally classified; a new classification into axonal ataxias, multiple system degenerations, and ataxic encephalopathies may be easier to relate to the pathophysiology.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2973644 TI - [3 cases of human syngamiasis in Guadeloupe]. AB - The authors report the 3 first cases of human syngamosis due to Mammomonogamus sp. in La Guadeloupe (French West Indies). After a review of the papers published on the previous 79 world-wide cases, they point out that this parasitic disease is restricted to the West Indies/South America area, and discuss the problem of the contamination mode; they claim that chronic cough is a constant signal symptom which suggests syngamosis in subjects who are native from, or have travelled in the endemic area. PMID- 2973645 TI - [Per-coelioscopic salpingectomy in extra-uterine pregnancy. A series of 25 cases]. AB - 25 cases of extra uterine pregnancy treated by salpingectomy via laparoscopy, representing 33 p. cent of all ectopic pregnancies reported between September 1985 and 1986. The authors describe the techniques, its advantages and its drawbacks. This technique represents an interesting alternative to classical laparotomy in the treatment of extra uterine pregnancy since it allows: a reduced surgical time and shortened hospital stay, a lesser risk of adhesions, an uneventful postoperative course, a good esthetic result. PMID- 2973646 TI - [Efficacy of oral contraceptives on acne. Apropos of a comparative study of Varnoline vs Diane in 69 women with acne]. AB - The authors conducted a comparative study of the effect of two oral contraceptives Varnoline and Diane in the treatment of androgenic manifestations: acne and hirsutism. The two products tested seem to have a similar efficacy on this type of clinical manifestations. PMID- 2973647 TI - Glycolytic enzyme activities in breast cancer metastases. AB - The activities of hexokinase, phosphofructokinase, aldolase, enolase and pyruvate kinase were studied in breast cancer metastases occurring at various sites and compared with the enzyme activities in a series of primary breast cancers. The activities of all enzymes studied were significantly higher in the metastases compared to the primary tumors (p less than or equal to 0.05). However, no changes in the isoenzyme patterns of enolase and pyruvate kinase were observed when the metastases were compared with primary breast cancers. Differences in location of the metastases did not lead to differences in enzyme activities. Our data suggest an association of an increasing rate of glycolysis with tumor progression. PMID- 2973649 TI - [Prevention of backache. Experience at a school of the back]. PMID- 2973648 TI - [Expression of pain in rheumatic disease. Exploratory study]. AB - An investigation done in 34 rheumatology departments on 259 patients in order to analyse their perception of pain and its repercussions on daily living. The main three diseases justifying hospitalization are: rheumatoid arthritis, lumbosciatica and bone neoplasia. In these patients, pain occurs daily in 75 p. cent of them and continuous in 32 p. cent; the intensity of the pain varies according to the time. There are repercussions on work (63 p. cent), walking (81 p. cent), but also on leisure (74 p. cent), friendly interelations (42 p. cent) and disposition which is disturbed in 62 p. cent of patients. Behavioral analysis in the presence of pain brings up differences depending on the patients: the elder the patient, the least they are able to respond to the handicap caused by pain. Analysis of the pain vocabulary suggest that if the terms expressing the experience of the patient are different according to the diseases, the usual description of painful phenomena are common to patients and the diseases in question. PMID- 2973650 TI - [Education and social rehabilitation of lumbalgic patients]. PMID- 2973653 TI - Suppressor activity as a composite property. PMID- 2973651 TI - [Treatment of lumbosciatica by intradisk injection of triamcinolone hexacetonide (hexatrione). Open study with 30 patients]. PMID- 2973652 TI - [Multicenter randomized double-blind study of triamcinolone hexacetonide versus chymopapain in the treatment of disk lumbosciatica. Initial results at 6 months]. PMID- 2973654 TI - Suppression needs a new hypothesis. PMID- 2973655 TI - Induction of basophilic differentiation in the human basophilic cell line KU812. AB - The potential for differentiation of the human basophilic leukaemia cell line KU812 was examined by means of a panel of physiologic and non-physiologic substances used as inducers. The phenotypic characteristics of non-induced KU812 cells included an immature morphology with scanty cytoplasmic granulation, expression of a low amount of high affinity, but no low affinity receptors (CD 23) for IgE, and a capacity for low-rate histamine synthesis. The differentiation process was characterized by a rapid (24 h) increase in histamine production a slower morphological maturation with the development of Alcian blue stainable granula demonstrable after 72 h. Concomitant with the phenotypic alterations, cell growth was inhibited. Differentiation in KU812 cells was inducible by Ara-C and to some extent by sodium butyrate, but not by dimethyl sulphoxide, retinoic acid, or gamma-interferon. Conditioned medium (CM) from cultured peripheral blood cells from atopic individuals and 18 out of 22 analysed glioma cell lines induced differentiation of the KU812 cells, whereas supernatant from only 1 out of 21 other cell lines, including carcinoma, melanoma, sarcoma, leukaemia, and normal fibroblasts had this activity. CM from the T-leukaemic cell line, Mo, also induced KU812 differentiation. A primary fractionation of the active substance from this cell line by reversed phase chromatography eluted the active substance at a concentration of 42-44% acetonitrile. Our present study has shown that the KU812 may serve as an appropriate model to study differentiation of basophils. In addition, its fast and specific response to biological factors makes it suitable as a biological assay for determination of active factor produced by atopic individuals. PMID- 2973656 TI - T- and B-cell functions in IgA-deficient patients. AB - In vitro lymphocyte function of 60 IgA-deficient patients (IgAdp) were studied. In the mitogen-induced lymphocyte activation test, peripheral blood mononuclear cells (PBMC) of IgAdp showed a weaker response to pokeweed mitogen (PWM) than those of controls (P less than 0.05), and the responses to phytohaemagglutinin (PHA) and concanavalin A (Con A) were normal. The amounts of IgA, IgG, and IgM secreted by PBMC from controls and IgAdp were measured in vitro by enzyme immunoassay. B cells were activated by PWM with or without hydrocortisone (HC) to inhibit HC-sensitive suppressor cells. Lymphocytes of IgA-deficient patients synthesized only minute amounts of IgA in vitro and the amounts of IgA synthesized correlated well with the serum IgA levels. Both IgG and IgM secretion by PWM-stimulated PBMC of IgAdp were also subnormal. The lymphocyte subset analysis in the peripheral blood revealed normal numbers and ratios of T cells, but IgAdp had reduced percentages of surface IgA-bearing cells. Co-culture experiments with isolated B cells, CD4- and CD8+ cells from IgAdp. and controls showed low B-cell capacity for IgA production as the most constant finding. Defects in T-cell functions or changes in the proportions of surface antigen carrying lymphocytes (other than IgA-) were infrequent and were not associated with any particular group of patients or type of IgAd (primary, acquired, or transient). PMID- 2973657 TI - The expression of CR1 and CR3 on non-modulated and modulated granulocytes of healthy blood donors as measured by flow cytofluorometry. AB - Complement receptors, CR1 and CR3, on neutrophils increase their cellular spontaneously at 37 degrees C or after mechanical stress during the cell preparation. We have established a cell preparation procedure and a cytofluorometric immunoassay method to evaluate the receptor expression in vivo in this study. The expression of CR1 and CR3 was studied after haemolysis in NH4Cl at different temperatures and incubation intervals. It was shown that cell preparation and receptor analysis must be performed at 15 degrees C or lower to avoid up-regulation of the receptor structures. Two minutes' incubation at 20 degrees C was sufficient to modulate the cells in this regard. Granulocytes from healthy blood donors were analysed and the mean fluorescence intensity (MFI), which reflects the number of receptors on the cell surface, showed a normal distribution for the CR1 (n = 158) and CR3 (n = 76) expression in the healthy population. The MFI of the two receptors showed a correlation (r = 0.71). Granulocytes from all donors could be modulated at 37 degrees C to a similar degree for both receptors (r = 0.76), despite the fact that they are supposed to be mobilized from different intracellular pools. A group of patients with localized inflammatory processes, such as sinusitis, differed from the healthy controls. In this group, the CR1 expression was not normally distributed, but 15 out of 26 patients (58%) had granulocytes with a CR1 expression exceeding the mean +/- SD of the normal population. PMID- 2973658 TI - Oestrogen and human T lymphocytes: presence of specific receptors in the T suppressor/cytotoxic subset. AB - Human spleen cells were investigated for specific oestrogen binding using oestradiol-17 beta. High affinity, low capacity binding was observed (Kd = 2.5 X 10(-10) M) and the characteristics of the oestrogen receptor identified were typical of this type of species. T lymphocyte subpopulations were prepared by negative selection by means of the monoclonal antibodies Leu 2a and 3a. Oestrogen receptors were only found in T cells of the suppressor/cytotoxic (CD8-positive) subset while helper/inducer lymphocytes (CD4-positive) showed no significant steroid binding by cytosol and nuclear assays. These results are supported by data from experiments into the effects of oestradiol-17 beta on immunoglobulin secretion, where it was apparent that only the suppressor/cytotoxic T-cell subpopulation was responsive to the presence of the steroid. PMID- 2973659 TI - IgG subclass deficiency in patients with Down's syndrome and aberrant hepatitis B vaccine response. AB - Seventeen adult patients with Down's syndrome (DS) and 19 adult healthy references were vaccinated with a hepatitis B vaccine in order to study the IgG subclass response. An enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies specific for IgG subclasses was employed. In spite of normal levels of total IgG1 and normal or even high levels of IgG3 in the DS patients, a significantly lower IgG1 response to the vaccine was observed in trisomic patients than in the references. PMID- 2973660 TI - An early hierarchic role of U1 small nuclear ribonucleoprotein in spliceosome assembly. AB - Splicing of nuclear precursor messenger RNA (pre-mRNA) occurs on a large ribonucleoprotein complex, the spliceosome. Several small nuclear ribonucleoproteins (snRNP's) are subunits of this complex that assembles on the pre-mRNA. Although the U1 snRNP is known to recognize the 5' splice site, its roles in spliceosome formation and splice site alignment have been unclear. A new affinity purification method for the spliceosome is described which has provided insight into the very early stages of spliceosome formation in a yeast in vitro splicing system. Surprisingly, the U1 snRNP initially recognizes sequences at or near both splice junctions in the intron. This interaction must occur before the other snRNP's (U2, U4, U5, and U6) can join the complex. The results suggest that interaction of the two splice site regions occurs at an early stage of spliceosome formation and is probably mediated by U1 snRNP and perhaps other factors. PMID- 2973661 TI - Lower cervical spondylosis and myelopathy in adults with Down's syndrome. AB - Abnormalities in the upper cervical spine resulting in cervical myelopathy in patients with Down's syndrome have been well-documented. However, two adult Down's syndrome patients recently presented with cervical myelopathy secondary to abnormalities of the lower cervical spine. Because of this, 105 Down's syndrome patients with normal upper cervical spines were evaluated clinically and radiographically. They were found to have an increased prevalence of lower cervical spondylosis that significantly correlated with physical findings consistent with cervical myelopathy. Therefore, physicians dealing with Down's patients should closely monitor neurological function and obtain flexion/extension laterals of the cervical spine to evaluate C1-C2 instability and degenerative changes in the lower cervical spine if a change in neurologic status is noted. PMID- 2973663 TI - Occupational asthma--time for scheduling as a compensable disease. PMID- 2973662 TI - Chromosome mapping of biological pathways by fluorescence-activated cell sorting and cell fusion: human interferon gamma receptor as a model system. AB - Human chromosome 6 encodes both the interferon gamma receptor as well as the class I major histocompatibility complex antigens, HLA-A, -B, and -C. However, the presence of chromosome 6 in somatic cell hybrids is insufficient to confer sensitivity to human interferon gamma (Hu-IFN-gamma) as assayed by class I HLA induction; it is necessary for both human chromosomes 6 and 21 to reside in the hybrid to generate a response to Hu-IFN-gamma. Treatment of such a hamster-human hybrid, Q72-18, with Hu-IFN-gamma induces the class I HLA antigens. Q72-18 cells selected by fluorescence-activated cell sorting for the loss of class I HLA induction also lost human chromosome 21. Fusions of such cells to a hybrid that contains only human chromosome 21 reconstitutes HLA antigen induction by Hu-IFN gamma. Furthermore, fusions of hybrids containing a translocated human chromosome 6q and the HLA-B7 gene to a line containing only human chromosome 21 or a translocated 21q also reconstitutes HLA-B7 mRNA and antigen induction by Hu-IFN gamma. Thus the segregation of cells on the basis of a biological effect by fluorescence-activated cell sorting and reconstitution by hybrid fusion provides a strategy by which some biological pathways can be mapped at a chromosomal level. PMID- 2973664 TI - Stomas. AB - The construction of intestinal stomas is a major part of a surgical procedure. A stoma should be formed by a surgeon who is not only technically skilled but also understands the potential metabolic and mechanical problems associated with an ileostomy or colostomy. Because of many of the complications are preventable, careful preoperative planning by the surgeon in conjunction with an enterostomal therapist is important to minimize the incidence of technical complications and to help prepare the patient psychologically for life with a stoma. When a complication does arise, it should be recognized promptly and dealt with appropriately. PMID- 2973665 TI - Is nocturnal asthma caused by changes in airway cholinergic activity? AB - A randomised, double blind, placebo controlled crossover trial of high dose nebulised ipratropium was carried out in 10 asthmatic patients with documented nocturnal bronchoconstriction. Patients received nebulised saline or ipratropium 1 mg at 10 pm and 2 am on two nights. Absolute peak flow (PEF) rates were higher throughout the night after the patients had received ipratropium (at 2 am, for example, mean (SEM) PEF was 353 after ipratropium and 285 l/min after placebo). The fall in PEF overnight, however, was similar with ipratropium and placebo. Patients were given a further 1 mg nebulised ipratropium at 6 am on both nights. There was a significant overnight fall in PEF on the ipratropium night even when comparisons were made between the times that maximal cholinergic blockade would be expected, PEF falling between 11.30 pm and 7.30 am from 429 to 369 l/min. The percentage increase in PEF, though not the absolute values, was greater after ipratropium at 6 am than at 10 pm. These results confirm that ipratropium raises PEF throughout the night in asthmatic patients, but suggest that nocturnal bronchoconstriction is not due solely to an increase in airway cholinergic activity at night. PMID- 2973666 TI - Lack of influence of low-dose acetylsalicylic acid (100 mg daily) on platelet survival time, beta-thromboglobulin and platelet factor 4 in patients with peripheral arterial occlusive disease. AB - In this study we investigated the influence of low-dose (100 mg daily) acetylsalicylic acid (ASA) on 111-In-platelet survival time (PST) and on plasma levels of beta-thromboglobulin (beta-TG) and platelet factor 4 (PF 4) in 30 patients (median age: 60 years) with arteriographically proven peripheral arterial occlusive disease in a chronic stable phase. We observed no significant changes of PST during therapy with ASA (weighted mean: 169.8----166 [median] hours; multiple hit: 168.3----170.6 hours), and also the plasma levels of beta-TG (median: 31.8----32.3 ng/ml) and of PF 4 (3.6----3.9 ng/ml) remained unchanged. PMID- 2973667 TI - Alloantibody and transferable suppressor activity induced by cyclosporine and blood transfusions in the rat. AB - The effect of cyclosporine on the alloantibody response to blood transfusion was investigated in inbred strains of rats by IHA and CELISA; recipient animals differed from the donors at the class I (RT1A) or both class I and class II (RT1B) antigens of the major histocompatibility complex. Alloantibody titers stimulated in high responder PVGu/c animals by blood transfusions were attenuated by cyclosporine; this effect was not demonstrated in low responder PVGc rats, as alloantibody titers decreased after further blood transfusions whether or not cyclosporine was given. Cyclosporine not only reduced the initial IgM response but suppressed the subsequent production of IgG. Splenocytes from rats receiving cyclosporine and blood transfusions from donors that differed from the recipients at the class I antigen were effective in suppressing the subsequent antibody response to blood transfusion. When blood transfusions from donors which differed from the recipients at both class I and class II antigenic loci were given after splenocyte transfer, a greater degree of immunosuppression was detected than if the transfusion donor differed only at the class I locus. These data suggest that the sensitization produced by blood transfusions and the persistence or decline of the alloantibody response depend upon the responder status of the recipient. Blood transfusions given with cyclosporine are capable of inducing suppressor activity that is transferable in spleen homogenates. Subsequent alloantibody responses are influenced by the class I and class II disparities of the donor and recipient animals. If these results can be extrapolated to clinical practice, cyclosporine should be given with pretransplant blood transfusions to prevent sensitization, and the transfusion donor should differ from the recipient at both class I and class II antigenic loci. PMID- 2973668 TI - Detrimental effect of chronic diabetes on growth and function of fetal islet isografts in mice. AB - We investigated (1), whether long-term (more than 6 months) streptozotocin induced diabetes in mice had a detrimental effect on the function of pancreatic islet isografts; and (2), whether there was an effect on graft function in chronically diabetic mice of continuous pretransplant insulin infusion. BALB/c female mice that had been diabetic for more than 6 months were each transplanted with 1/2 of a 17-day fetal mouse pancreas that had been in organ culture for 14 days. All animals were grafted with same batch of tissue. One group of animals received continuous intraperitoneal infusion of regular insulin via an Alzet 2002 osmotic pump at the rate of 0.5 U/day for 14 days prior to grafting. Matched, chronically diabetic animals with pumps containing diluent alone, acutely diabetic animals of the same age, and acutely diabetic younger animals were used as controls. At 20 weeks after transplantation the grafts were removed and their insulin content measured. Following transplantation and removal of the pumps, all acutely diabetic animals returned to euglycemia within 6 weeks. The chronically diabetic animals which received diluent alone took 11 weeks to reach euglycemia compared to 7 weeks for their littermates that had received insulin. Graft insulin content was decreased from 16,300 +/- 4100 ng in the acutely diabetic animals to 9600 +/- 5200 ng in the chronically diabetic, non--insulin-treated group. The chronically insulin-treated group, however, had grafts with 16,400 +/- 5100 ng. Our studies suggest that there is a detrimental effect of chronic diabetes on graft insulin content that is ameliorated by pretransplant insulin therapy. PMID- 2973669 TI - Therapeutic effect of 15-deoxyspergualin on acute graft rejection detected by 31P nuclear magnetic resonance spectrography, and its effect on rat heart transplantation. AB - We investigated the effect of 15-deoxyspergualin (DSG) on graft rejection, starting administration at the onset of rejection and on the induction of immunologic unresponsiveness. Hearts from WKAH rats were transplanted into the neck of ACI rats. The energy metabolism of the grafted hearts was followed by 31P nuclear magnetic resonance spectroscopy. The day that energy metabolism started to fall was defined as the onset of rejection, and intraperitoneal administration of DSG was initiated at 5 mg/kg/day for 15 days from this day. The grafted heart arrested in 2 of 10 rats 9 and 11 days after transplantation, respectively, but the remaining 8 recovered from rejection and 5 of them showed evidence of immunologic unresponsiveness. Of 10 rats treated with DSG from the day of transplantation, only 1 rat showed evidence of unresponsiveness. The initiation of DSG treatment from the onset of rejection resulted in a higher percentage of induction of unresponsiveness. Therefore, DSG was considered to specifically inhibit lymphocyte clone expansion at the onset of rejection. Spleen cells obtained from recipients 7-10 days after the end of DSG treatment were administered to syngeneic ACI rats grafted with WKAH hearts. Graft survival was significantly prolonged, but long-term unresponsiveness could not be transferred. However, immunologic unresponsiveness could be adoptively transferred in 3 of 5 rats receiving spleen cells from syngeneic rats that had recovered from rejection after DSG treatment and had acquired long-term unresponsiveness. These results suggest that suppressor cells are resistant to DSG and are spared and participate in the maintenance of immunologic unresponsiveness. PMID- 2973670 TI - Lack of secondary suppressor response in enhanced allografted rats with endogenous suppressor activity in spleen and diminished cytotoxic T cell response. AB - In order to better understand the mechanisms of alloimmunization occurring in transplantation, we studied the immune response of an inbred rat strain (WF) to varied presentation of alloantigens (LEW). Hyperimmunized rats acutely reject transplanted allografts and demonstrate direct cytotoxic activity, as well as an expanded potential to generate cytotoxic and suppressor cell activity. In contrast, passively and actively enhanced animals, which exhibit allograft tolerance, demonstrate endogenous suppressor cell activity and significantly reduced potential to generate cytotoxic and suppressor cell activity. Thus, the method of immunization of the same responder strain animals with identical alloantigens will determine whether rejection or tolerance is evidenced following transplantation. The suppressor T cell activity demonstrated in the enhanced animals appears to regulate the immune response by reducing cell proliferation when challenged with alloantigen. However, when restimulated in vitro with donor cells, splenic suppressor activity declines--i.e., there is not a secondary suppressor response. PMID- 2973671 TI - Abrogation of tissue immunogenicity by immunotoxin treatment of human fetal pancreas. PMID- 2973672 TI - Pyomyositis of the anterior abdominal wall. PMID- 2973673 TI - [Heteromorphism of chromosome 1, 9 and 16 homologs in persons living in regions differing in the level of longevity]. AB - Regional and age-related peculiarities of chromosomal polymorphism are established as a result of studies in C-band heteromorphism of chromosomes 1, 9 and 16 in the long-lived subjects, their relatives and population groups of the Abkhaz and Ukrainian Republics. Heteromorphism frequencies of chromosomes 1 and 9 homologs are higher in the Abkhaz as compared with the Ukrainian Republic. Age related differences as to the degree of expression of chromosome 9 C-band heteromorphism are found: in the Abkhaz Province the frequency of variants with a high heteromorphism degree increases with age, while in the Ukrainian one--with a low heteromorphism degree. PMID- 2973674 TI - Seasonal prevalence of protostrongylid and Dictyocaulus species of lungworms in Awassi sheep in north-west Syria. PMID- 2973675 TI - Clinical signs following experimental lungworm infection and natural bovine respiratory syncytial virus infection in calves. AB - Similar clinical signs have been reported in calves infected either by Dictyocaulus viviparus or bovine respiratory syncytial virus. Three experiments were carried out to establish the clinical picture and the course of the disease in animals with these infections. The clinical signs of calves infected with lungworm included coughing, nasal discharge, tachypnoea, abdominal breathing and pyrexia, and auscultation of their lungs revealed increased bronchial sounds. Similar signs were also observed after infection with bovine respiratory syncytial virus, but the signs were more acute and resolved more rapidly than in animals infected with lungworm larvae. Calves infected with lungworm had more serious clinical signs after infection with bovine respiratory syncytial virus than calves, which were not infected with lungworm. PMID- 2973677 TI - Ivermectin therapy and degradation of cattle faeces. PMID- 2973676 TI - Immunity to parasitic bronchitis of yearling cattle treated with ivermectin during their first grazing season. AB - A group of 12 winter-born calves was divided into two groups of six. During the following summer one group grazed on pasture infected with Dictyocaulus viviparus, and was treated with ivermectin injections at three, eight and 13 weeks after turn out. The other group remained housed. Both groups were housed during the winter and then together with a group of younger calves were challenged with a trickle infection of D viviparus larvae at the rate of 25 third stage larvae/kg bodyweight for one month and then slaughtered. The group which had been exposed to previous infection was least affected by parasitic bronchitis and on the basis of serological titres and worm burdens had developed resistance to the challenge infection. The other older group was also more resistant than the younger calves. PMID- 2973679 TI - [Medical support for the Soviet troops in the Kursk offensive operation]. PMID- 2973678 TI - Intestinal permeability to macromolecules during colibacillosis in piglets. AB - Intestinal macromolecular permeability to macromolecules was determined in a group of specific pathogen free piglets before and after they were infected experimentally with Escherichia coli. Six hours after the infection all piglets developed a profuse diarrhoea. The urea and total protein concentrations in the serum increased markedly after the onset of diarrhoea. Haemoglobin concentration and PCV decreased steadily during the experiment but blood glucose concentration and lipid composition of the faeces did not change. No structural abnormalities in the jejunal and ileal mucosa were seen. No uptake of macromolecules (40,000 KDa) was found suggesting that molecules with a molecular weight of 40,000 or more do not play a role in the persistent diarrhoea sometimes seen in piglets after colibacillosis. PMID- 2973680 TI - [Troop medical supplies during the battle on the Kursk arch]. PMID- 2973681 TI - [Military patriotic training at the institute]. PMID- 2973682 TI - [Holter monitoring in assessing the adequacy of the health resort treatment of patients with ischemic heart disease and rhythm disorder]. PMID- 2973683 TI - [Effect of sanatorium treatment on the clinico-biochemical indices of children with peptic ulcer]. PMID- 2973684 TI - [Effectiveness of health resort climatic treatment including oxylidin electrophoresis and the correction of the climatic adaptation of patients with the sequelae of viral encephalitis]. PMID- 2973686 TI - [Metabolic status of the liver mitochondria in rats drinking water containing various forms of iodine]. PMID- 2973685 TI - [Specialized neurocardiology departments at a sanatorium--a new form of patient treatment organization]. PMID- 2973687 TI - [Bishofit as a balneotherapy factor]. PMID- 2973688 TI - [Multi-year observations of the humoral immunity status of patients with tick borne encephalitis in 3 endemic regions]. AB - Analysis of the results of the laboratory diagnosis of tick-borne encephalitis (TBE) by HI test over many years proved a statistically significant decrease by the 80's in the intensity of humoral immunity in the subjects with a history of tick-borne encephalitis in Kemerovo Province, Udmurtia and Latvia. A decrease in antihemagglutinin levels in the blood of convalescents after TBE was detected in endemic foci located in different geographical regions differing by epidemiological parameters; it did not depend on the portion of infected ticks and ratio of the clinical forms of the disease in a focus. Parallel tests by HI of the diagnostic value of several antigens of TBE virus strains isolated from different sources in different regions with sera from TBE patients revealed no differences among them. The antigenic activity of the strains in HI test did not depend on the source and region of isolation (Kemerovo Province, Udmurtia and Latvia). PMID- 2973689 TI - Pregnancy-associated osteoporosis. PMID- 2973690 TI - [The appearance of specific IgG and IgA antibodies to Streptococcus pneumoniae in serum and lung lavage fluid of mice after the administration of different antigens. 2. IgG and IgA antibody detection after intraperitoneal, aerosol and oral immunization with Streptococcus pneumoniae]. AB - The stimulation of secretory and systemic IgG- and IgA-antibodies in the lung lavage fluid was examined in an animal test. It was also examined in the serum of mice after once to four times oral, aerosol or intraperitoneal immunization with total formalin destroyed bacteria of the species Streptococcus pneumoniae type I. Not only in the lung lavage fluid, but also in the sera specific antibodies could be found with the enzyme immunoassay. An intraperitoneal antigen application mainly resulted in increase of the specific IgG in the serum, whereas an oral and especially aerosol antigen application mainly developed a stimulation of IgA in the lung lavage fluid. PMID- 2973691 TI - [Ischemic reaction in coronary angioplasty (PTCA) as a prerequisite for improving left ventricular function following intravenous thrombolysis in acute myocardial infarct]. AB - The incidence of ischemic ECG-changes and anginal symptoms during PTCA and the influence of PTCA on left ventricular function were studied in a consecutive series of 40 patients with recanalized infarct artery and significant (greater than 50%) single vessel disease. PTCA was performed within the first week (4.1 +/ 2.5 days) after intravenous streptokinase (1.5 Mio U i.v. over 60 min) in acute myocardial infarction (duration of pain less than four hours). The primary success rate of PTCA was 78% (31/40). In 25 of 31 (81%) successfully dilated patients, there were ischemic ECG-changes (ST-segment elevation greater than 0.1 mV), and in 14 patients anginal symptoms were observed during balloon inflation. Follow-up angiography after six months showed restenosis or reocclusion in 5/22 (23%) of the patients with an ischemic reaction, and in 3/6 (50%) without ischemic signs during PTCA. Only patients with an ischemic reaction and without restenosis demonstrated an increase in left ventricular ejection fraction (p less than 0.05) and regional wall motion in the infarct region (p less than 0.01), in contrast to patients with restenosis or without an ischemic reaction during angioplasty. Thus, after intravenous streptokinase an ischemic reaction can be detected in a high percentage (81%) as an indicator of salvaged myocardium. Significant improvement of left ventricular function was only demonstrable in patients with ischemic signs during PTCA without restenosis. PMID- 2973692 TI - [Increased primary success rate in coronary angioplasty using a guide wire with attached balloon (microprobe)]. AB - In 15 cases of 350 consecutive coronary angioplasties conventional low profile balloon catheters did not traverse the stenosis or occlusion over the guide-wire. A balloon on a wire device ("Microprobe", 2 mm) passed the stenosis or occlusion easily with a good primary result in 14 cases. The angioplasty procedure could then be completed with a larger standard balloon catheter. Use of the balloon on a wire device expands the technical facilities of angioplasty and increases the primary success rate in technically difficult cases. PMID- 2973693 TI - Immunogenicity of a heat-treated recombinant DNA hepatitis B vaccine. AB - The effect of heating on the reactogenicity and the immunogenicity of a recombinant DNA hepatitis B vaccine was examined. Treatment of this vaccine for one week at 37 degrees C did not significantly alter these properties when compared to vaccine stored at 4 degrees C. These results have important implications in countries where facilities for the transport and handling of vaccines are often inadequate. PMID- 2973694 TI - Persistence of antibodies following vaccination with a yeast-derived recombinant hepatitis B vaccine. AB - The persistence of anti-HBs antibodies elicited in response to vaccination with a recombinant yeast-derived hepatitis B vaccine was examined. Three years after vaccination following two different schedules (0, 1, 6 months and 0, 1, 2, 12 months), anti-HBs antibody titres obtained after vaccination with the yeast derived vaccine were similar to those observed using a plasma-derived vaccine. At this time, nearly 100% of all vaccinees had anti-HBs antibody titres over the protective limit of 10 mIU ml-1. These results suggest that the yeast-derived vaccine will confer protection against hepatitis B infection for about the same length of time as the plasma-derived vaccine. PMID- 2973696 TI - [Prevention of chronic staphylococcal skin diseases]. PMID- 2973695 TI - [Immunologic indices of patients with occupational dermatoses caused by organophosphate pesticides]. PMID- 2973697 TI - [History of the development of the social-prophylactic trend in the control of skin and venereal diseases in the Armenian SSR]. PMID- 2973698 TI - Determination of ergosterol as a measure of fungal growth using Si 60 HPLC. AB - In order to determine to fungal growth of Fusarium graminearum 480, a method was developed for the extraction and estimation of ergosterol, a sterol specific for fungi. This method includes the direct saponification of bound ergosterol to fungal mycelia followed by n-hexane extraction and quantification using. High performance liquid chromatography (HPLC) with UV-detection. This procedure proved to be superior compared with other methods, since the yield of ergosterol yields was higher (up to 40%). n-Hexane extracts contained minor impurities which interfered with the UV-detection and the retention time of the compound was halved using Si 60 HPLC. The protein and ergosterol contents in F. graminearum cultures increased proportionally over a 3-week incubation period. The fungal formation of the mycotoxin zearalenone started at a level of 50 mg/kg ergosterol and increased rapidly in the stationary phase of growth, which was characterized by decreasing rates of ergosterol formation. PMID- 2973699 TI - Restoration of the rat nasal septum and turbinates after damage by intranasal electrocoagulation. PMID- 2973700 TI - [A general decline in caries among the youthful population in the GDR?]. PMID- 2973701 TI - [Effect of the degree of polymerization and particle size of the polymer powder on the processing and properties of denture synthetics]. PMID- 2973702 TI - [Differentiated age-adjusted oral hygiene in preschoolers]. PMID- 2973703 TI - [Value of conventional and newer measurements (modified tensor analysis) in the teleradiograph exemplified by Class-II therapy]. PMID- 2973704 TI - [The sum of the angles as a growth indicator of the facial skull. A geometric study]. PMID- 2973705 TI - [The growth pattern in cleft lip and palate patients with reference to familial disposition]. PMID- 2973706 TI - [Oxidative metabolism of the peripheral neutrophilic granulocytes in patients with rapidly progressing and with localized juvenile periodontitis]. PMID- 2973707 TI - [Functional defects of the local granulocytes in periodontal diseases]. PMID- 2973708 TI - [Sugar intake of adults and children in Erfurt]. PMID- 2973710 TI - [Uptake of radioactive orthophosphate (32PO4) by Streptococcus mutans OMZ 176]. PMID- 2973709 TI - [Comparative materials testing of human bone and Kallocryl Cf]. PMID- 2973711 TI - [Biomorphs of oral acellular organic deposits]. PMID- 2973713 TI - Unsatisfactory sensitivity and specificity of indirect immunofluorescence tests for the presence or absence of antibodies to chicken anaemia agent (CAA) in sera of SPF and broiler breeder chickens. PMID- 2973712 TI - Susceptibility of Ureaplasma urealyticum to ten chemotherapeutic agents. AB - Tetracyclines and erythromycin are the chemotherapeutic agents most often used in the Federal Republic of Germany for eradication of U. urealyticum from the male urogenital tract. Few data on current susceptibility in the FRG are available. Therefore, we investigated the minimal inhibitory concentration (MIC) values of 10 chemotherapeutic agents against 27 isolates from the male urethra and 4 ATCC strains of U. urealyticum by a micro broth dilution method using a modified U-9 medium. The MIC90 values (microgram/ml) of the chemotherapeutic agents tested were as follows: minocycline-0.25, doxycycline-1.0, tetracycline and chlortetracycline-2.0, erythromycin and streptomycin-4.0, chloramphenicol, gentamicin, rosoxacin-8.0, spectinomycin-32.0. According to the blood levels attained in treatment, the chemotherapeutic agents can be divided into three groups based on the MIC90-values, i.e. sensitive: chloramphenicol, doxycycline, minocycline and streptomycin; weakly effective: chlortetracycline, tetracycline, spectinomycin and erythromycin; and resistant: gentamicin and rosoxacin. PMID- 2973714 TI - [Late observations in a familial foci of hepatitis B using a full set of specific markers of the infection]. AB - The complete set of specific markers of hepatitis B has been identified, thus making it possible to evaluate the spread of this infection in family foci, as well as the intensity and dynamics of the epidemic process, under the conditions of prolonged observations. The study has shown that the spread of hepatitis B infection is determined by the presence of HBeAg in antigen-positive patients with viral hepatitis B and HBsAg carriers. PMID- 2973715 TI - [Epidemiologic problems of pneumococcal infection]. AB - During the microbiological examination of 314 tentatively healthy donors, residents of Leningrad, Streptococcus pneumoniae was detected in the rhinopharynx of 94 donors (29.9%) in amounts ranging from a few individual microbial cells to 10(6) microbial cells with the mean geometric concentration equal to 3.58 lg. Antipneumococcal antibodies were detected in the blood of 147 donors (46.8%). The occurrence of antipneumococcal antibodies was approximately 2.5 times higher than the average level of carriership, but in most cases the dynamics of immunological reactions changed in the same direction. Considerable monthly and annual fluctuations in the occurrence of pneumococcal carriership and antipneumococcal antibodies, as well as their correlation with the epidemic rises of acute respiratory diseases, influenza and other acute respiratory viral diseases, were established. These data indicate that pneumococcal infection plays a definite role in the epidemic rises of acute respiratory diseases. PMID- 2973716 TI - [Epidemiologic effectiveness of mass revaccination against measles]. AB - The data on the epidemiological effectiveness of the mass booster immunization of students of high schools, as well as professional and technical schools, against measles are presented. Booster immunization is considered to be a highly effective antiepidemic measure aimed at decreasing morbidity rate in measles. PMID- 2973717 TI - [History of the Department of Psychiatry of the Leningrad Pediatric Medical Institute (on the 50th anniversary of its founding)]. PMID- 2973718 TI - [Ontogenetic aspect of sensory-perceptive disorders in mental diseases]. AB - Based of study of 1595 files of patients with various mental diseases ranging from preschool to senile age groups, an analysis is presented of age peculiarities of perceptive and psychosensory disorders, perception distortions in depersonalization-derealization symptoms, illusions, hallucinations. The incidence of different kinds of sensory-perceptive disorders in a number of mental diseases is collated. Ontogenetic features of sensory and perceptive disorders are juxtaposed with these functions' normal ontogenesis. PMID- 2973719 TI - [Early alcoholization and drug abuse behavior as a manifestation of delinquency in children and adolescents]. AB - Long-term investigation of 115 children and adolescents of either sex with delinquent behavior is presented. Of these, in 81 an early alcoholization and in 29 drug abuse behavior has been revealed. The first experience with toxic substances and alcohol was generally inspired by persons of the same age. Of wines, spirits prevailed, and of toxic substances gasoline and acetone-containing stuff. The level and direction of social activity, especially at leisure, were major prognostic factors. Twenty-four percent of patients had a steady level of social adaptation with their signs of the disease satisfactorily compensated. PMID- 2973720 TI - Nonsecretory myeloma: a study on hypoimmunoglobulinemia. AB - In evaluating hypoimmunoglobulinemia in a patient with nonsecretory myeloma, studies of in vitro immunoglobulin synthesis were performed during a 3-year period. Impaired polyclonal response to mitogen stimulation associated with excessive suppressor activity was demonstrable at diagnosis. Despite successful chemotherapy, serum immunoglobulin levels remained depressed with a persistent impairment in polyclonal immunoglobulin synthesis but without evidence for excessive suppression. In nonsecretory myeloma, as in secretory myeloma, a defective B-cell response to differentiation signals may be a contributory mechanism in the hypoimmunoglobulinemia seen in this disease. PMID- 2973721 TI - Dichloromethylene biphosphonate retards femoral expansion in normal and castrated adult male rats. AB - This study reports the effects of dichloromethylene biphosphonate (Cl2MBP), an inhibitor of bone remodeling, on femoral expansion in four groups of adult male rats; (1) sham-operated controls; (2) sham operated + injections of Cl2MBP; (3) castrated (osteoporotic), and (4) castrated + injections of Cl2MBP. After controlling for body weight, analyses of covariance revealed significant differences in total femoral width between animals that had received Cl2MBP and those that had not. The results indicated that Cl2MBP treatment retarded femoral expansion in both castrated and normal adult male rats. PMID- 2973722 TI - Elevation of tear fluid plasmin in corneal disease. AB - Plasmin concentration was determined in tear fluid from 76 eyes showing corneal epithelial disorders, such as corneal ulcers and erosions due to trauma or contact lens wear. Nearly 70% of the eyes with corneal disease had plasmin in their tear fluid, whereas plasmin was present in only 20% of the eyes in a control group of 50 eyes. Re-examination of the plasmin positive control eyes revealed conjunctival bacterial growth or mild subclinical viral infection in most cases. We conclude that plasmin is released into the tear fluid in the presence of corneal or conjunctival lesions or infections, suggesting a pathogenic role of plasmin in these disorders. The significance of the occurrence of plasmin in tear fluid during corneal wound healing is discussed. PMID- 2973723 TI - Immunoreactive opsin and glial fibrillary acidic protein in persistent hyperplastic primary vitreous. AB - An 8-month-old boy had an anterior type of persistent hyperplastic primary vitreous in the right eye. Results of needle biopsy, performed because of elevated intraocular pressure, disclosed clusters of blastic cells. The eye was enucleated on the suspicion of retinoblastoma. Histological examination showed retrolental fibrovascular tissue and retinal dysplasia. Immunoreactive opsin was detected in the innermost structures and in photoreceptor-like cells of rosettes. We conclude that photoreceptor cells differentiated to express opsin, even when neighbouring cells were abnormally arranged. An immunocytochemical study of glial fibrillary acidic protein demonstrated glial proliferation in the inner layer of the retina but not in the preretinal space. PMID- 2973724 TI - Influence of ramipril on release of norepinephrine during sympathetic nerve stimulation in isolated rabbit hearts. PMID- 2973725 TI - Levodopa addiction. A case study. AB - A case is presented of a young woman with a serious addiction to levodopa who over the years developed an extrapyramidal syndrome and chronic paranoid psychotic behaviour. The possible pathophysiological mechanism is discussed. PMID- 2973726 TI - Psychiatric aspects of Down's syndrome. AB - In a survey study of psychiatric morbidity, based on a representative sample from the Danish mental retardation register, 44 adults with Down's syndrome (DS) were compared with 258 other mentally retarded adults. Assessed by the parameters psychiatric disorder, behaviour problems, neurotic traits, and deviant social interaction, the DS group functioned better on all parameters. However, male and female DS patients were very different, the females constituting a superior well functioning group while the males had major problems in every area. High prevalence rates of dementia and infantile autism were found in the DS group. General function rapidly decreased with age in DS patients. PMID- 2973727 TI - Carotid cavernous fistula associated with Fogarty catheter angioplasty. PMID- 2973728 TI - [Functional capacity of T cell subpopulations in mice. II. Effect of allogeneically stimulated T-cell subpopulations on the primary MLR]. AB - The effect of allogeneically stimulated lymphocytes or their subpopulations respectively on a primary allogeneic MLC was studied. It was shown that this effect depends either on the amount of the transferred cells or on the composition of the culture medium. In contradiction to the common view that allogeneically stimulated lymphocytes act as suppressor cells, a significant stimulating effect, mediated by these cells, could be demonstrated under respective conditions. Also in the experiments with the subpopulations of allogeneically stimulated lymphocytes the effects of these cells are rather influenced by the conditions then determined by a restricted functional repertoire of the cells. PMID- 2973730 TI - Transumbilical balloon coarctation angioplasty. PMID- 2973729 TI - Transductal balloon angioplasty for coarctation of the aorta in the neonate: preliminary observations. AB - Transductal balloon angioplasty for coarctation of the aorta is described wherein the balloon dilatation catheter is positioned across the coarctation after having passed into the descending aorta from the main pulmonary artery via a patient ductus arteriosus. This procedure was performed in two neonates with excellent immediate clinical results and objective improvement in the pressure gradient. The mechanism of relief of obstruction appears to be ductal dilatation, which allows bypassing the posterolateral shelf of aortic coarctation. This technique is recommended in neonatal coarctations with open ductus arteriosus, especially if the femoral artery cannot be catheterized for performing conventional transfermoral balloon coarctation angioplasty. PMID- 2973731 TI - Bidirectional precordial ST-T wave changes in pure right ventricular ischemia. PMID- 2973732 TI - Percutaneous coronary laser angioplasty: successful clinical application of a new thermal cap catheter coaxially guided over a steerable central guide wire. PMID- 2973733 TI - Percutaneous peripheral laser angioplasty: demonstration of the clinical safety and efficacy of a new coaxial-guided laser-heated cap system. PMID- 2973734 TI - Left ventricular structural characteristics in unilateral renovascular hypertension and primary aldosteronism. AB - To assess the importance of the renin-angiotensin system and plasma volume as determinants of hypertensive left ventricular hypertrophy and its anatomy, patients with unilateral renovascular hypertension and primary aldosteronism were studied by echocardiography. Blood pressure, age and sex were matched as closely as possible. The 19 patients with unilateral renovascular hypertension and the 19 patients with primary aldosteronism were similar in age, sex and blood pressure (168 +/- 19/97 +/- 11 and 163 +/- 17/99 +/- 10 mm Hg, respectively), but plasma volume was increased in the patients with primary aldosteronism. Interventricular septal thickness, left ventricular posterior wall thickness, left ventricular mass index and relative wall thickness did not differ between the 2 groups of patients. There was a significant correlation between the level of systolic blood pressure and either left ventricular mass index (r = 0.34, p less than 0.05) or relative wall thickness (r = 0.58, p less than 0.001) in both groups of patients. Left ventricular end-diastolic dimension index was increased in the patients with primary aldosteronism compared with those with unilateral renovascular hypertension (3.2 +/- 0.4 vs 2.9 +/- 0.3 cm/m2, p less than 0.02). When confined to the patients with systolic pressure greater than or equal to 150 mm Hg, relative wall thickness was significantly increased in the patients with unilateral renovascular hypertension. Patients with primary aldosteronism and unilateral renovascular hypertension of similar blood pressure levels, age and sex have almost identical degrees of left ventricular hypertrophy and anatomy. In contrast, the patients with primary oldosteronism had increased left ventricular dimension index.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973736 TI - Long-term outcome of patients with asymptomatic restenosis after percutaneous transluminal coronary angioplasty. PMID- 2973735 TI - Identification of best electrocardiographic leads for diagnosing left ventricular hypertrophy by statistical analysis of body surface potential maps. AB - In view of the increased risk of cardiovascular mortality associated with left ventricular (LV) hypertrophy, early recognition and quantitation of LV hypertrophy are important clinical goals. The standard 12-lead electrocardiogram is the easiest and most widely used noninvasive method for the diagnosis of LV hypertrophy; unfortunately, the diagnostic accuracy of commonly used electrocardiographic criteria remains unsatisfactory. Body surface potential maps contain diagnostic information not present in conventional lead systems. The present investigation combines the increased information content of surface maps with the power of multivariate statistical techniques in order to identify practical subsets of electrocardiographic leads that would allow improved diagnosis of LV hypertrophy. Discriminant analysis was performed on 120-lead data simultaneously recorded in 250 normal subjects and 214 patients with LV hypertrophy using as features instantaneous voltages on time-normalized P, PR, QRS and ST-T waveforms as well as the duration of these waveforms. Leads and features for optimal separation of 173 normal subjects aged greater than or equal to 30 years from 122 patients with pure LV hypertrophy were selected. A total of 6 features from 5 torso sites accounted for a specificity of 97% and a sensitivity of 94%. The single most potent discriminator was the duration of the P wave; voltages were measured in mid and late P on leads located in the lower left parasternal area, the left precordial region and the upper right back, in mid-QRS on a lead positioned 10 cm below V1 and slightly before the peak of the T wave on a lead in the lower left flank.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973738 TI - Release of atrial natriuretic factor after pericardiectomy for chronic constrictive pericarditis. PMID- 2973737 TI - Comparison among acquired immune deficiency syndrome patients with and without clinical evidence of cardiac disease. PMID- 2973739 TI - A symposium: Interventional cardiology at a crossroad--diagnostics and therapeutics. March 26, 1988, Atlanta, Georgia. PMID- 2973740 TI - The Maryland experience: angioplasty and valvuloplasty using percutaneous cardiopulmonary support. AB - Vessel closure during coronary angioplasty in patients with poor ventricular function or extensive distribution of the target vessel, or both, can result in hemodynamic collapse--a potentially fatal complication. A similar situation can develop as a result of problems arising during aortic valvuloplasty. In an attempt to extend the benefits of angioplasty and valvuloplasty to patients with more advanced coronary and valvular heart disease, semipercutaneous cardiopulmonary circulatory support has been used as an adjunct to these interventions. In our initial experience with supported angioplasty/valvuloplasty, 11 vessels in 9 patients and 6 aortic valves were successfully dilated. Complete cardiopulmonary support was achieved in all cases, permitting prolonged balloon inflations; intraprocedural complications were limited to vessel injury and blood loss. One patient each died after angioplasty and valvuloplasty. Symptomatic improvement was observed in all surviving patients. PMID- 2973741 TI - Primary percutaneous coronary angioplasty: a protagonist's view. AB - The wide variety of reperfusion strategies now available has led to confusion regarding the optimal therapy of patients with acute myocardial infarction. In assessing the comparative efficacy of reperfusion strategies, safety factors, recanalization rates, reocclusion rates, myocardial salvage and improvement in survival must be compared. In this analysis, primary percutaneous transluminal coronary angioplasty (PTCA) compares favorably to intravenous thrombolytic therapy or combination PTCA and thrombolytic therapy as a reperfusion strategy. Primary PTCA has not been reported to cause intracerebral hemorrhage, whereas thrombolytic strategies are associated with an incidence of a 0.2 to 1.4% rate of intracerebral bleeding. Strategies using PTCA have recanalization rates of only 33 to 75%. Disappointingly, a major failure of all reperfusion strategies (including primary PTCA) is a high rate of reocclusion and reinfarction. Primary PTCA is associated with a 7 to 16% increase in global ejection fraction. Similar improvement in ejection fraction has not been demonstrated in patients treated with thrombolytic therapy alone. Although in-hospital mortality is improved, a substantial 1-year mortality persists for intravenous thrombolytic therapy. Long term survival is greater when PTCA therapy is used. In addition, PTCA does favorably improve survival in high-risk patients with cardiogenic shock. Although it is unlikely that most patients with acute myocardial infarction will have primary PTCA as a treatment option, this therapy remains extremely attractive for centers where primary PTCA is readily available. PMID- 2973742 TI - Percutaneous transluminal coronary angioplasty: the second decade. AB - Over the last decade, percutaneous transluminal coronary angioplasty has become a major therapeutic technique of myocardial revascularization. Advances in catheter design and operator experience have made angioplasty a viable alternative for many patients with both single- and multivessel disease who would otherwise require bypass surgery. Acute closure and restenosis remain the 2 principal limitations of angioplasty. Means of controlling these problems, including intracoronary stenting and antiplatelet and thrombolytic agents, are now being studied. Furthermore, controlled prospective randomized trials are underway to assess the value of angioplasty compared with coronary bypass surgery in the treatment of multivessel disease. These trials will help to establish the limits of coronary angioplasty and its relative advantages in terms of therapeutic results and costs. PMID- 2973743 TI - The uncertain role of thrombolytic angioplasty within the treatment strategy for myocardial infarction. AB - The logical hypothesis that early angioplasty after lytic therapy would be of considerable clinical value flows from the recognized shortcomings of pharmacologic reperfusion efforts. These shortcomings are: (1) failure to lyse some thrombotic occlusions; (2) limited quantifiable salvage relative to risk area, possibly related to low magnitude or reperfusion coronary flow through a tight residual stenosis; (3) frequent early postlytic reocclusion and reinfarction; and (4) common postinfarction angina after lysis and some degree of salvage. Recently however, important controlled clinical trials of percutaneous transluminal coronary angioplasty (PTCA) after lytic therapy (most notably Thrombolysis and Angioplasty in Myocardial Infarction study group, Thrombolysis in Myocardial Infarction trial and the European Cooperative trial) have not confirmed this hypothesis. Very early PTCA after intravenous lytic therapy has not produced a favorable short-term outcome compared with delayed PTCA or a more conservative approach. Where is the flaw? Is it the hypothesis, the patient selection criteria or the specifics of therapeutic algorithms? While these issues are further investigated, current prudent clinical recommendations are best modified downward from enthusiastic rapid postlytic dilatation. PMID- 2973744 TI - Direct bonding of orthodontic brackets to porcelain veneer laminates. AB - The forces required to debond orthodontic attachments from porcelain veneer laminates were studied in vitro. Brackets were bonded to 160 veneered bovine incisor teeth before the determination of the debond force. The independent variables studied were resin type, priming agent, porcelain surface preparation, and debonding time. The average debond forces were compared with those obtained by debonding brackets bonded to natural teeth by means of the acid-etch technique. The bond between the resin and the porcelain surface was found to be satisfactory for direct bonding of orthodontic attachments. Roughening the porcelain surface and using a silane primer required an average debond force comparable to that of the acid-etched enamel bond at 24 hours. However, it increased the risk for porcelain fracture during debonding. Roughened surfaces and surfaces with micro-fractures could be satisfactorily finished and polished with either a series of graded Ceramiste points or a diamond-impregnated polishing wheel followed by a diamond polishing paste. PMID- 2973745 TI - Growth studies in infants and children with Down's syndrome and elevated levels of thyrotropin. AB - A retrospective survey of 147 patients with Down's syndrome (age range, 4 months to 27 years) showed that 60% had a thyrotropin (TSH) level higher than 5.7 mU/L in the presence of high or normal thyroxine levels. The remaining 40% of the group had low to normal TSH values. High TSH levels were predominant in patients under 4 years of age (94 children), ie, during the phase of active growth, and showed a declining trend with increasing age. All 94 infants had delayed growth of all parameters including head circumference, height, and weight, as compared with normal infants, and growth was particularly retarded in patients with TSH levels greater than 5.7 mU/L. Thyroid dysfunction, expressed as a high TSH concentration, is associated with growth retardation in children with Down's syndrome who are younger than 4 years. PMID- 2973746 TI - Components and modifiers of the healthy worker effect: evidence from three occupational cohorts and implications for industrial compensation. AB - The authors examined the components and modifiers of the healthy worker effect using mortality data from three occupational cohorts: the employees of Atomic Energy of Canada Limited followed between 1950 and 1981, a 10% sample of the Canadian labor force followed between 1965 and 1979, and workers at the Eldorado Resources Limited Beaverlodge uranium mine followed between 1950 and 1980. Two important components of the healthy worker effect have been identified in these cohorts, namely, initial selection of and continuing employment of healthy individuals. There is less evidence for a contribution from the existence of differential risk factors among employed individuals as compared with the general population. The healthy worker effect is, however, substantially modified by time since employment, sex, age, specific cause of death, and specific occupation. Because of this variation, it is inappropriate to account for the healthy worker effect by a single parameter, and all of the above factors must be taken into account in any appropriate analysis. When the only available comparison group for an occupational cohort is the general population, the healthy worker effect is unlikely to have any substantial influence on the process of assessing causality for any observed association or attributing cause in an individual case. This would be particularly true for cancer, and even more so for lung cancer, a disease often associated with industrial compensation cases. PMID- 2973747 TI - Atrial natriuretic factor in renal failure and posthemodialytic postural hypotension. AB - Atrial natriuretic factor (ANF) levels were ten times normal in hemodialysis patients before dialysis. ANF was not cleared by the dialyzer membrane but plasma levels decreased 47% by the end of dialysis. Patients undergoing peritoneal dialysis had plasma ANF levels four times normal and had detectable ANF in their dialysate. Hemodialysis patients with a marked fall in BP after dialysis had higher ANF levels (P less than 0.05) and lower norepinephrine (NE) levels (P less than 0.05) associated with a failure to increase NE in response to dialysis. Elevated ANF levels are associated with postdialysis hypotension in hemodialysis patients. PMID- 2973748 TI - Insulin, insulin-like growth factors, and vascular endothelium. AB - Endothelial cells form the intimal lining of the entire vascular system. The vascular endothelium is continuously and directly bathed by components of the bloodstream and represents the initial fixed anatomical surface with which these components come in contact. In the past decade, the methodologies for studying endothelial cell functions have markedly advanced, enabling direct and detailed study of the vascular endothelium. From such studies, it is now apparent that the vascular endothelium represents an extraordinarily complex network of cells demonstrating a multitude of distinct anatomic, metabolic, and immunologic properties critical to such processes as angiogenesis, atherosclerosis, thrombosis, neoplasia, and a variety of metabolic disorders including homocystinuria and diabetes mellitus. This report will focus on the interactions of insulin and the insulin-like growth factors (IGFs) with vascular endothelium, based on studies with cultured endothelial cells, isolated microvessels, and perfused organ systems. Data will be presented relevant to the following concepts: (1) endothelial cells, in culture and in vivo, have specific receptors for insulin, IGF-I, and IGF-II; (2) insulin, IGF-I, and IGF-II have both distinct and overlapping functions in cultured endothelial cells; (3) cultured endothelial cells process receptor-bound insulin, IGF-I, and IGF-II, by distinct processes; (4) in vivo, capillary endothelial receptors are integrally involved in the transport of intact insulin to subendothelial sites of insulin action; and (5) vascular endothelium has specialized cellular features that are likely to contribute to the unique interactions of endothelial cells with insulin and the IGFs. PMID- 2973749 TI - Evaluation of patients with back pain of suspected inflammatory nature. AB - PURPOSE: Detection of early inflammatory back disease is often difficult. Certain clinical characteristics have been reported to increase the likelihood of its detection in referral patients, but the usefulness of these clinical characteristics has not been evaluated in an open population. In our study, we undertook to evaluate the value of the clinical history as a screening test for inflammatory back disease in a general population. PATIENTS AND METHODS: Twenty three male patients with back pain of moderate duration and with clinical characteristics suggestive of inflammatory back disease were recruited by advertising and were studied by various means, including computed tomography (CT), scintigraphy, and radiography. RESULTS: One patient had radiographic sacroiliitis. Two had positive results for the B27 antigen, and another had positive results for the cross-reacting HLA antigen B7. Eight patients had abnormal scintiscans of the sacroiliac joints. Twenty-one of 23 patients and 20 of 23 control subjects had abnormalities that were detected by CT. Repeat plain radiographs of the pelvis done 36 months after enrollment into the study did not uncover further evidence of sacroiliitis. CONCLUSIONS: These results indicate plain radiographic evidence of sacroiliitis will often not develop in patients with historical features suggestive of inflammatory back disease even with long term evaluation, thus vitiating the specificity of these historical findings in men with back pain of relatively brief duration. PMID- 2973750 TI - Calcium control of smooth muscle contractility. AB - Ca2+ is a primary second messenger that binds to an intracellular receptor protein, calmodulin. Increases in cytosolic Ca2+ concentration mediated by activation of cell surface receptors result in the formation of a Ca2+ calmodulin complex that regulates many Ca2+-dependent cellular processes. In smooth muscle, Ca2+/calmodulin activates myosin light chain kinase, which phosphorylates the regulatory light chain of myosin. This phosphorylation reaction increases the actin-activated MgATPase activity of myosin and is associated with increases in contractile properties, including force, stiffness, and maximal shortening velocity. These biochemical and biomechanical responses occur rapidly (seconds) in response to physiological stimulation involving neurotransmitter activation of smooth muscle cells. Thus, the Ca2+-dependent phosphorylation of the myosin light chain is a primary event in activation of smooth muscle contraction. PMID- 2973751 TI - Evolution of laboratory parameters during sickle cell painful crisis: evidence compatible with dense red cell sequestration without thrombosis. AB - We find that during 51 episodes of sickle cell painful crisis indirect bilirubin fell 52% from its steady state value of 2.3 +/- 1.9 mg% to a value of 1.1 +/- 0.37 mg% at the end of crisis (p less than .00000085). The indirect bilirubin decline correlates with a decrease in the dense sickle cells during crisis (r = .31, p less than .0009). During steady state, both indirect bilirubin and lactic acid dehydrogenase correlate significantly with number of dense red cells (r = .62, p less than .000002 and r = .32, p less than .02 respectively). Platelet counts, beta-thromboglobulin, Platelet Factor 4, and Fibrinopeptide A levels all were elevated during steady state and did not change during the evolution of crisis. These data demonstrate that elevated indices usually associated with platelet activation are a feature of the steady state of sickle cell disease but argue against thrombosis as a factor in the progression of a sickle cell painful crisis episode. The parallel decline of both dense cells and bilirubin during painful crises indicates that the disappearance of dense cells during crisis is not caused by hemolysis and supports the hypothesis that dense red cell sequestration, in the absence of evidence of thrombosis, is an intrinsic component of the evolution of sickle cell painful crisis. PMID- 2973753 TI - Health care groups put on push for free hepatitis B vaccine. PMID- 2973752 TI - Prostatic carcinoma with lung metastases in a former bagasse worker; LH-RH analogs as an alternative therapy. PMID- 2973754 TI - Premenstrual hyperphagia: reprise. PMID- 2973755 TI - Efficacy of trimetrexate, a potent lipid-soluble antifolate, in the treatment of rodent Pneumocystis carinii pneumonia. AB - Trimetrexate is a lipid-soluble antifolate that has been shown in vitro to be a much more potent inhibitor of Pneumocystis carinii dihydrofolate reductase than the conventionally used inhibitor trimethoprim. To evaluate the in vivo efficacy of trimetrexate, steroid-treated rats which spontaneously develop P. carinii pneumonia were used. Rats treated with trimetrexate (25 mg/kg/d) plus sulfamethoxazole (250 mg/kg/d) orally responded at least as well as rats treated with trimethoprim (50 mg/kg/d) plus sulfamethoxazole. Trimetrexate alone administered orally was ineffective in treating P. carinii infection, but subcutaneous (sc) trimetrexate (7 mg/kg/d) significantly decreased the intensity of infection compared to controls. Trimetrexate is a potent antifolate that may provide an effective alternative to pentamidine and trimethoprim-sulfamethoxazole for treatment of P. carinii pneumonia in humans. PMID- 2973756 TI - Failure of a dengue 1 sub-unit vaccine to protect mice against a lethal dengue virus infection. AB - Mice immunized with DEN-1 pre-matrix protein produced antibody which reacted with dengue virus infected cells and intact virions but failed to neutralize virus in vitro or in vivo. Newborn mice passively immunized with anti-DEN-1 pre-matrix antibody also failed to survive intracerebral infection with 100 LD50 of any dengue virus serotype. PMID- 2973757 TI - [The role of the hirsute score in identifying a group at risk of disorders of reproductive function]. PMID- 2973758 TI - High thoracic epidural sufentanil for post-thoracotomy pain: influence of epinephrine as an adjuvant--a double blind study. PMID- 2973759 TI - Atrial natriuretic factor may mediate the renal effects of PEEP ventilation. AB - Mechanical ventilation with PEEP decreases urine output and urinary sodium excretion. Observed changes in cardiac output, renal blood flow, renin release, and antidiuretic hormone (ADH) secretion do not adequately explain the renal effects of PEEP. Altered release of atrial natriuretic factor (ANF), which is natriuretic and diuretic, may complete this explanation. The following hypothesis was tested: a PEEP-induced decrease in transmural right atrial pressure decreases ANF release, and this mechanism mediates subsequent alterations in renal function. Seven female mongrel dogs were anesthetized with halothane and their lungs ventilated mechanically for three consecutive 40 min periods of 0 PEEP, 10 cmH2O PEEP, and 0 PEEP. Addition of 10 cmH2O PEEP during mechanical ventilation decreased right atrial dimension and transmural right atrial pressure while intracavitary right atrial pressure was increased. Urine output was significantly decreased during PEEP, as were absolute and fractional excretion of sodium and osmolar clearance. PEEP ventilation resulted in a consistent and significant decline in plasma ANF concentration (82 +/- 11 to 62 +/- 11 pg/ml, P less than 0.05). Hemodynamic parameters, renal function, and ANF concentration returned to control values after cessation of PEEP. A second series of experiments in five dogs demonstrated a close temporal relationship between changes in atrial dimension or atrial transmural pressure, plasma ANF concentration, and urine output or sodium excretion. The results of this study demonstrate that PEEP induced decreases in atrial distension resulted in decreased ANF release, which may mediate, in part, the antinatriuretic and antidiuretic effects of PEEP. PMID- 2973761 TI - Effect of exercise on atrial natriuretic peptide (ANP) levels in patients with COPD. PMID- 2973760 TI - Hepatitis B vaccination programs for hospital workers: results of a statewide survey. AB - To assess the implementation of hepatitis B virus (HBV) vaccination programs for hospital workers, we mailed questionnaires to all 229 licensed Michigan hospitals. The response rate was 96% (221/229); of these, 68% (150/221) had vaccination programs. Although multiple hospital characteristics were associated with the presence of a vaccination program, characteristics that independently predicted the presence of a program were medical school affiliation, nonpsychiatric specialty, and the existence of a hepatitis B immune globulin protocol. The most common reason given (56%, 40/71) for the absence of a program was insufficient worker risk of hepatitis B infection; this response was frequent in psychiatric (91%, 10/11) and rural hospitals (61%, 11/18). Among high-risk workers, attending physicians were less likely than other high-risk workers to be included in vaccination programs (68% vs. 95%, respectively). Fear of vaccine associated acquired immunodeficiency syndrome was most frequently cited as the primary reason for vaccine refusal. We conclude that unwarranted fears about the vaccine's safety need to be dispelled, that high-risk physicians should be included in vaccination programs, and that rural and psychiatric hospital policies reflect their perceived risk of occupational HBV infection. PMID- 2973763 TI - Four years of experience with ultrasound-guided follicle aspiration. PMID- 2973762 TI - Coronary angioplasty for acute myocardial infarction. AB - PURPOSE: To critically review the role of coronary angioplasty for acute myocardial infarction. DATA IDENTIFICATION: Studies published from January 1982 to June 1988 were identified through a search of the English-language literature using MEDLINE and thorough extensive hand searching of bibliographies of identified articles. STUDY SELECTION: All consecutive patient series and randomized trials of coronary angioplasty for acute myocardial infarction. DATA EXTRACTION: Demographic characteristics, technical results, clinical outcomes, and left ventricular function results were collated. RESULTS OF DATA SYNTHESIS: Three randomized trials have compared immediate and deferred coronary angioplasty after intravenous tissue plasminogen activator. Immediate angioplasty was associated with increased mortality, increased need for emergency bypass surgery, higher transfusion requirement, and no benefit in terms of left ventricular functional recovery. CONCLUSIONS: The role of coronary angioplasty for myocardial infarction has thus far been established as adjunctive, in a delayed time frame after thrombolytic therapy. Coronary angioplasty has not been shown to reduce in hospital mortality or improve resting left ventricular function. A deferred strategy has improved exercise left ventricular performance, and several follow up studies suggest a favorable long-term prognosis. PMID- 2973765 TI - [Dermohypodermitis induced by subcutaneous aminoglycosides. Allergic or toxic mechanism?]. PMID- 2973764 TI - Luteinizing-hormone-releasing hormone agonist treatment in patients with previously failed folliculogenesis during in vitro fertilization therapy. PMID- 2973766 TI - [Synovitis-acne-pustulosis-hyperostosis-osteitis (SAPHO) syndrome]. PMID- 2973767 TI - Reevaluation of the "glycolytic complex" in muscle: a multitechnique approach using trout white muscle. AB - Preliminary characterization of the "glycolytic complex," formed in trout white muscle, revealed that phosphofructokinase (PFK) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) are bound to particulate matter largely by ionic interactions; increasing neutral salt or charged metabolite concentrations released bound PFK and GAPDH. GAPDH was consistently solubilized at lower salt concentrations, indicating that it is not bound as tightly as PFK, but both enzymes were readily solubilized at physiological concentrations of salts and metabolites. pH titrations indicated that PFK binding is dependent on group(s) with a pKa of 7.3 in 30 mM imidazole. PFK binding increased at lower pH values; at 150 mM KCl the apparent pKa value is 6.5. Experiments with polyethylene glycol 8000 (PEG), which is used to mimic the high in vivo protein concentrations under in vitro conditions, showed that the binding of PFK and GAPDH increased with increasing PEG concentrations. Interestingly, at 5% PEG, only the PFK binding response depended on the ionic composition of the medium--with increased binding occurring at the pH of the exhausted muscle and decreased binding at control pH values. These results suggested that only PFK reversibly bound to cellular structures in response to changing conditions and disagrees with previous studies showing binding of several glycolytic enzymes as measured using the dilution method (F. M. Clarke, F.D. Shaw, and D.J. Morton (1980) Biochem. J. 186, 105 109). In order to determine whether artifactual binding was measured by the dilution method, two new methodologies were employed to measure enzyme binding in vivo: (a) whole muscle slices were pressed to quickly extrude cellular juice, and (b) muscle strips were finely minced and centrifuged to liberate cytoplasmic contents. Both methods indicated that, under physiological conditions, up to 70% of the total cellular phosphofructokinase may be bound, but other glycolytic enzymes are bound to a lesser extent (10-30%). This result contrasts those obtained with the dilution method, and suggests that dilution of cellular contents may result in an overestimation of the percentage of enzyme associated with cellular structures; this is dramatically shown for glyceraldehyde-3 phosphate dehydrogenase. The viability of the glycolytic complex in trout white muscle is discussed in light of the decreased binding measured using these new methodologies. PMID- 2973768 TI - Selective inhibition of chlorophyll biosynthesis by nicotinamide. AB - Rhodobacter sphaeroides grown in the presence of nicotinamide excreted bacteriochlorophyll precursors, 2,4-divinyl protochlorophyllide (DV-Pchlide) and a small amount of 2-monovinyl protochlorophyllide (MV-Pchlide). Accumulation of these pigments indicates that nicotinamide inhibited the bacteriochlorophyll biosynthetic pathway site-specifically between DV-Pchlide and MV-Pchlide. This phenomenon is also observed in an aerobic photosynthetic bacterium, Erythrobacter sp. OCh 114. Among 12 nicotinamide derivatives and isomers tested, only nicotinamide was effective, indicating that in addition to the completeness of the pyridine ring skeleton at positions 1 to 3, the carboxylic acid amide group is essential for this inhibition. The technique described in this report permits the simple preparation of large quantities of DV-Pchlide. PMID- 2973769 TI - [Chemo-endocrine therapy of advanced breast cancer]. AB - There have been few randomized trials that have shown a significantly longer survival after chemo-endocrine therapy than the chemotherapy or the endocrine therapy alone, although there seem to be rationales for simultaneous combination of the both modalities. Some new strategies that would make the response rate higher as well as obtain a longer survival by means of chemo-endocrine therapy was suggested. At the present time, sequential endocrine therapy to chemotherapy is recommended for the advanced or recurrent breast cancer patients, except for those with larger tumor burden, liver metastasis, rapid growing tumors, and ER negative tumors. PMID- 2973770 TI - Circulating levels of sialic acid and glycosaminoglycans: a diagnostic test for ankylosing spondylitis. AB - The circulating levels of sialic acid (N-acetylneuraminic acid) and glycosaminoglycans (GAGs) were measured in 69 patients with spinal disorders of orthopaedic interest (ankylosing spondylitis 17, osteofluorosis 6, idiopathic backache 10, osteoarthrosis 16, osteoporosis 20). The serum GAG levels showed no statistically significant change from control values in the five disorders investigated in the present study. Although osteoporosis and osteoarthrosis showed a decrease in serum sialic acid (SA) levels, the mean ratio (SA/GAG) demonstrated no change from control values. Idiopathic backache showed no difference in any of the parameters studied when compared with control values. Ankylosing spondylitis and osteofluorosis had a remarkable similarity in their clinical and radiological features, but a divergent mean value of ratio was noted. The mean ratio of both the conditions also showed a statistically significant difference from the control value. This suggests that the SA/GAG ratio can be used as a diagnostic test in ankylosing spondylitis. PMID- 2973772 TI - Morbidity and mortality of coronary bypass surgery in patients 75 years of age or older. AB - To determine factors associated with an increased risk of postoperative complications in elderly patients, 60 consecutive patients 75 years of age or older undergoing isolated coronary artery bypass grafting (CABG) were evaluated. Thirty-nine patients (65.0%) had at least one major postoperative complication, including 2 deaths (3.3%). Low body weight was the only univariate predictor (p less than 0.05) of an increased likelihood of complications overall. Prior cardiac operation, low serum cholesterol value, and prolonged cardiopulmonary bypass time were associated with increased bleeding. Electrocardiographic evidence of left ventricular hypertrophy was associated with prolonged postoperative confusion. Age of 80 years or more and increased cross-clamp time were predictive of pulmonary dysfunction. Low cardiac output occurred more frequently in patients with nonsinus rhythm, prior cardiac operation, recent congestive heart failure, or elevated level of blood urea nitrogen. Identification of risk factors for specific complications should prompt further studies to define ways of reducing morbidity and the resultant high cost associated with CABG in elderly patients. PMID- 2973771 TI - In vitro cell-mediated immunity after thermal injury is not impaired. Density gradient purification of mononuclear cells is associated with spurious (artifactual) immunosuppression. AB - Mononuclear cells isolated by density gradient centrifugation from the peripheral blood of burn patients, but not healthy volunteers, are contaminated with large numbers of nonmononuclear cells. These contaminating leukocytes could cause artifactual alterations in standard in vitro tests of lymphocyte function. Thus, we compared the in vitro blastogenic response of density gradient purified leukocytes and T-cell purified lymphocytes from 13 burn patients to mitogenic (PHA) and antigenic stimuli. The mitogenic and antigenic response of the patients' density gradient purified leukocytes were impaired compared to healthy volunteers (p less than 0.01). However, when the contaminating nonlymphocytes were removed, the patients' cells responded normally to both stimuli. Thus, density gradient purified mononuclear cells from burn patients are contaminated by leukocytes that are not phenotypically or functionally lymphocytes. Since the lymphocytes from burn patients respond normally to PHA and alloantigens after the contaminating nonlymphocyte cell population has been removed, it appears that in vitro assays of lymphocyte function using density gradient purified leukocytes may give spurious results. PMID- 2973773 TI - The changing mortality of myocardial revascularization: coronary artery bypass and angioplasty. AB - The risk factors and outcome for the first 150 consecutive patients undergoing coronary artery bypass grafting (CABG) in 1985 (CABG '85) were compared with those of the first 150 patients undergoing CABG in 1975 (CABG '75) and those of the first 150 patients to have percutaneous transluminal coronary angioplasty (PTCA) in 1985 (PTCA '85). The CABG '85 patients had a significantly higher (p less than 0.05) incidence of known operative risk factors including advanced age, female sex, severity of angina, history of recent infarction, triple-vessel disease, left ventricular dysfunction, and emergency operation than the CABG '75 cohort. The clinical profile of the PTCA '85 patients closely resembled the low risk profile found in the CABG '75 patients. Overall mortality following CABG more than doubled during the decade studied (3% versus 7%, p = 0.07). This study suggests that the increased mortality associated with CABG in 1985 is due in part to the inclusion of more high-risk patients in the surgical population. In addition, the application of PTCA removes low-risk patients from the surgical candidate pool and adds more patients requiring emergency operations, thereby further contributing to the overall decline in the clinical status of patients referred for operation. PMID- 2973774 TI - [Antagonistic and insecticidal properties of soil micromycetes]. AB - It was shown that microscopic fungi isolated in plague foci had an antagonistic effect on the plague causative agent. In the majority of the cases it was associated with their insecticidal activity against plague-transmitting fleas. It is expedient to consider the relations revealed between the burrow biocenosis components in investigation of plague enzootic aspects and development of new biological insecticides for control of the infection carriers. PMID- 2973775 TI - Myasthenia gravis after general anesthesia and hepatitis B vaccine. AB - A 48-year-old man presented with the first symptoms of myasthenia gravis one month after a general anesthesia and a second dose of hepatitis B plasma vaccine. Whether either event may have acted as a nonspecific challenge to the patient's immune system is speculative, but the case is described to discover similar observations, if any. PMID- 2973776 TI - [Structural and ultrastructural aspects of normal and atrophic nasal mucosa]. PMID- 2973777 TI - [Dose-response relation of intravenous enoximone in congestive cardiac insufficiency]. AB - Enoximone (MDL 17043) is a new generation inotropic drug which acts by inhibiting phosphodiesterase and is endowed with both inotropic and vasodilator properties. The purpose of this study, which involved 23 patients aged from 18 to 75 years in NYHA class III or IV and with evidence of severe haemodynamic disturbances (cardiac index below 2.5 1/mn/m2, pulmonary wedge pressure above 15 mmHg), was to evaluate the acute haemodynamic responses to doses of enoximone that ranged from 0.25 to 2.50 mg/kg administered by bolus intravenous injection. Heart failure was either of ischaemic origin (6 cases) or idiopathic (10 cases) or due to various causes (7 cases). Group A patients (n = 11) received the drug in low doses (less than or equal to 1 mg/kg) as opposed to group B patients (n = 12) who were given high doses (greater than 1 mg/kg). Results were evaluated from the amplitude and duration of the haemodynamic response at maximum effect time (30 min). The following parameters were measured: cardiac index, pulmonary wedge pressure, systemic vascular resistance, mean arterial pressure and heart rate. Cardiac index and pulmonary wedge pressure were significantly improved in both groups (P less than 0.005): cardiac index +39 p. 100 in group A, +55 p. 100 in group B; pulmonary wedge pressure -36 p. 100 in group A, -48 p. 100 in group B; systemic vascular resistance -46 p. 100 in group B. Heart rate and arterial pressure were not significantly altered. The duration of response was 1 to 3 hours in group A patients and 4 to 8 hours in group B patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973778 TI - [Usefulness of neurophysiologic technics in the evaluation of lumbosciatica]. PMID- 2973780 TI - [Immunoreactivity in ischemic heart disease of atherosclerotic etiology]. AB - Using a combined immunomorphologic and clinico-immunologic approach to the analysis of section and clinical findings, the authors introduce a specific nosological form of coronary heart disease in which there is a cause-and-effect relation implying the development of sensitization to atherogenic lipoproteins, rapid advance of coronary atherosclerosis and the onset of coronary insufficiency. PMID- 2973781 TI - [Architectonic characteristics of the heart ventricles in a perimembranous defect of the interventricular septum]. AB - The results of quantitative morphological macro-, micro-, and ultrastructural (myocardial biopsies) studies of the ventricles with perimembranous septal defect in 135 infants under the age of one year have been compared. In newborns there appeared ventricular myocardial hyperplasia and low absolute and relative values of all the myocardial parameters versus normal control. At the age of 3 months the parameters were similar to or above the control ones exceeding them considerably by the age of 10-12 months. The comparison of the parameters of the septal parts showed predominant abnormality of the sinus part indicating that the perimembranous ventricular septal defect entity is a sinus one. PMID- 2973779 TI - Investigation of the role of glycans for the biological activity of Semliki Forest virus grown in Aedes albopictus cells using inhibitors of asparagine linked oligosaccharides trimming. AB - The effects of N-linked-oligosaccharide-processing inhibitors on the formation of Semliki Forest virus (SFV) in C6/36 Aedes albopictus cells were investigated. The glycosidase inhibitors deoxynojirimycin, deoxymannojirimycin and swainsonine prevented the formation of Endo-H resistant structures, but had little effect on virus formation and on the biological activities of the virus. Tunicamycin greatly inhibited virus formation, but had little effect on cell-cell fusion from within and the cleavage of p 62. These results indicate that correct glycosylation is not a prerequisite for biological activities of SFV, whereas glycosylation per se is needed for virus production. PMID- 2973782 TI - [Characteristics of myocardial hypertrophy in arterial hypertension of various origins and in atherosclerosis]. AB - Sets of conventional macroscopic and up-to-date histochemical techniques were employed to study hearts from 267 patients who had died of essential hypertension, atherosclerosis or secondary renal hypertension. Excessive cardiac mass was found disease-specific and related to the time since hypertension onset. Cardiac hypertrophy was augmenting with growing deficiency of coronary blood supply and showed specific correlation between cardiomyocyte nucleus and cytoplasm within each nosological unit mentioned. This may serve evidence for the absence of a common morphofunctional underground for this process. Essential hypertension is characterized by stepwise nuclear changes in cardiomyocytes relevant to advancing hypertrophy. PMID- 2973783 TI - Interaction of lipoprotein Lp(a) and low density lipoprotein with glycosaminoglycans from human aorta. AB - The lipoprotein complexing activity of glycosaminoglycans (GAG) prepared from human aortas with lipoprotein Lp(a) in comparison to low density lipoproteins (LDL) was determined tubidimetrically in the presence of Ca++. In control experiments, purified chondroitin-6 sulfate and proteoglycans (PG) were used. Lp(a) exhibited approximately a threefold higher reactivity. Analyzing the chemical composition of the complexes, we found that Lp(a) had greater than fourfold higher binding capacity for GAG. The binding capacity of Lp(a) to PG was 3.4-fold higher as compared to LDL. The binding capacity of both lipoproteins for chondroitin-6 sulfate was only 50% in comparison to GAG, but again Lp(a) was four times more reactive. Neuraminidase treatment of LDL or Lp(a) did not interfere with GAG or chondroitin-6 sulfate binding. If, on the other hand, Lp(a) was treated with dithiothreitol and the Lp(a)-specific protein (apoprotein [apo] a) was removed, the GAG binding was reduced by about 45%. Apo a by itself gave no insoluble complexes with GAG. LDL and Lp(a)-s GAG and -LP(a)-PG complexes were incubated with mouse peritoneal macrophages (MPM), and the stimulation of cholesteryl ester formation was studied. At identical lipoprotein cholesterol concentrations, Lp(a)-GAG complexes exhibited a 1.3-fold higher stimulation of cholesterol esterification as compared to LDL-GAG. This difference was even more striking if lipoproteins were compared at a molar basis. PG-lipoprotein complexes were much more active with respect to interactions with MPM. The highest amount of cholesterol ester formation upon incubation with MPM was found with PG-Lp(a) complexes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2973784 TI - Persistence of commercial modified live reovirus vaccines in chicks. AB - Four modified live reovirus vaccines were compared with a field isolate (81-176) of reovirus for safety in 1-day-old and 1-wk-old specific-pathogen-free chicks. At 1, 3, 5, and 7 wks after vaccination, blood, cloacal swabs, and hock tendon samples were collected and assayed for residual virus. Tendon samples were also examined microscopically and scored for histological changes. Vaccine efficacy was monitored by challenging chicks 7 wks postvaccination via the footpad with the 81-176 isolate. Serum samples were collected throughout the trials and assayed for reovirus antibodies using an enzyme-linked immunosorbent assay. In chicks vaccinated at 1 day of age, three of the four vaccine viruses were consistently recovered from the hock tendon tissues. Post-challenge virus recoveries were similarly positive for the same vaccine viruses. Histological lesion scores of the tendons paralleled the degree of virus recovery. In the chicks vaccinated at 1 wk old, none of the vaccine viruses were recovered, nor were tendon lesion scores substantially elevated. Following challenge, however, virus was recovered from all vaccinated groups. PMID- 2973785 TI - DNA sequence dependence of ATP hydrolysis by RecA protein. AB - The DNA sequence dependence of the ATPase activity of RecA protein has been investigated for a variety of single strand octamer and hexadecamer homopolymers and alternating copolymers. Under assay conditions where the single strand DNA concentration exceeds the RecA protein concentration, significant differences in the rates of ATP hydrolysis for the various single strand DNA oligomer cofactors are observed. Under the conditions examined, the order of efficiency of the DNA cofactors in inducing RecA mediated ATPase activity is found to be: dA16 greater than dT16 greater than d(TC)16 greater than dT8 greater than dC16 greater than dA8 = dG8 greater than dG16 greater than dC8 greater than d(AG)16. These results demonstrate not only a dependence of RecA ATPase activity on the sequence composition of short single strand DNA they further reveal ATPase activity can be affected by the nearest neighbor nucleotide sequence of short DNA cofactors. PMID- 2973786 TI - Classification of the insulin-like growth factor binding proteins into three distinct categories according to their binding specificities. AB - Competitive binding experiments with insulin-like growth factor (IGF)-1, IGF-2 and des-(1-3)-IGF-1 have confirmed the interpretation based on limited amino terminal sequence analysis that at least three types of IGF binding protein occur. In addition to the acid stable subunit of the large serum binding protein which exhibits des-(1-3)-IGF-1 binding only slightly less than IGF-1, the small IGF binding proteins can be separated into two classes based on differences in des-(1-3)-IGF-1 and IGF-2 binding potencies. PMID- 2973787 TI - CSF-1 and C-FMS gene expression in human carcinoma cell lines. AB - The macrophage-specific colony stimulating factor CSF-1 is required for the growth and differentiation of monocytes. The cell surface receptor for CSF-1 is identical to the product of the c-fms proto-oncogene. The present studies have monitored CSF-1 and c-fms expression in human carcinoma cell lines. Two of three human ovarian carcinoma cell-lines expressed multiple species of CSF-1 mRNA. Furthermore, detection of CSF-1 transcripts was associated with secretion of CSF 1 protein that was increased after phorbol ester treatment. CSF-1 mRNA was also detectable in 4 breast and 2 lung carcinoma cell lines. In contrast, c-fms expression was found only in SK-Br-3 breast carcinoma cells. Similar studies in 2 human choriocarcinoma cell lines demonstrated the presence of c-fms, but not CSF 1, transcripts. While phorbol ester treatment was associated with increased c-fms mRNA levels in choriocarcinoma cells, this agent had no effect on CSF-1 expression. These findings indicate that: 1) CSF-1 expression is frequent in human ovarian, breast and lung carcinoma cells; and 2) coexpression of the CSF-1 and c-fms genes, as found in monocytes is infrequent in malignant epithelial and choriocarcinoma cell lines. PMID- 2973788 TI - Pharmacokinetics of nisoldipine. III. Biotransformation of nisoldipine in rat, dog, monkey, and man. AB - After intraduodenal administration of 14C-labelled (+/-) 3-isobutyl-5-methyl 1,4 dihydro-2,6-dimethyl-4-(2-nitrophenyl)-pyridine-3,5-dicarboxylate (nisoldipine, Bay k 5552) to rats approx. 68% of the dose was excreted in the bile in the first 6 h. In an isolated perfused rat liver model the excretion with the bile was 56% of the total dose within 3 h. The recovery of radioactivity from orally administered [14C] nisoldipine was approx. 32% (rat), 23% (dog), 73% (monkey) and 74% (man), resp., in the urine. The unchanged drug was neither detected in the urine nor in the bile, but nisoldipine was present in plasma of the rat 30 min after dosing and up to 24 h in man. The drug was extensively metabolized: 18 biotransformation products were identified by comparison with synthetic reference compounds using combined GC-MS, 1 NMR-spectroscopy, mass spectrometry, gas chromatography/radio-gas chromatography and two-dimensional thin layer chromatography, 6 of them being quantitatively important (about 80% of the radioactivity excreted in urine). The metabolites identified accounted for approx. 82% (rat: bile and urine), 19% (dog, due to the low renal excretion), 58% (monkey: urine) and 64% (man: urine) of the excreted dose, resp. The following biotransformation steps occurred: hydroxylation of the isobutyl moiety, dehydrogenation of the 1,4-dihydropyridine system, oxidative ester cleavage, hydroxylation of one of the methyl groups in 2- or 6-position and subsequent oxidation to the carboxylic acid, oxidation of one of the methyl groups of the isobutyl moiety to the carboxyl group reduction of the aromatic nitro group (minor biotransformation reaction) and glucuronidation as phase II reaction. PMID- 2973789 TI - [Epidemiologic surveillance of tetanus in Rio Grande do Sul, Brazil]. PMID- 2973791 TI - Hemodynamic and atrial natriuretic peptide responses to fluid removal and reinfusion in hemodialysis patients. AB - The authors have studied the sequential changes in hemodynamic responses and Atrial Natriuretic Peptide (ANP) levels following bicarbonate hemodialysis with fluid removal followed by an infusion of saline and then ultrafiltration of the added fluid. Cardiac output, stroke volume, and thoracic fluid index (TFI) were measured by transthoracic impedence cardiography. Left cardiac work index (LCWI) was derived using these hemodynamic and mean arterial pressure (MAP) measurements. MAP was lower following dialysis when compared to pure ultrafiltration of a similar fluid loss. LCWI decreased during both dialysis and ultrafiltration. TFI, which is inversely related to intrathoracic fluid, increased following dialysis and ultrafiltration. ANP levels fell following fluid removal but remained elevated above normal control levels. The authors conclude that ANP levels are related to fluid not cardiac status in dialysis patients and that impedence cardiography is of value in determining hemodynamic and fluid changes during dialysis. PMID- 2973790 TI - Is atrial natriuretic polypeptide a cause of pathophysiology in total artificial heart animals? AB - The purpose of this study was to clarify whether plasma ANP levels (p-ANP) are abnormal in TAH animals, and if ANP plays an important role in circulatory failure in TAH animals. In five TAH goats that survived for 51-171 days, changes in hemodynamic parameters, plasma levels of renin, angiotensin I and II, aldosterone (p-RAA), and p-ANP were measured, and correlations between p-ANP and other parameters were studied, and histologic study of ANP and assay of the ANP content in atrial tissue was undertaken. Generally, p-ANP of TAH goats fell temporarily after surgery but then recovered to preoperative levels. Histopathologic studies of atrial tissue and ANP granules proved almost normal, although the coronary blood supply was occluded for more than 170 days; the ANP content in the atrium did not show significant changes after 51 days of TAH pumping. In one case prominent correlations between p-ANP and p-RAA were observed, and in another case significant correlation between p-ANP and arterial pressure was observed. However, these correlations were not observed in every case. In TAH goats the plasma ANP level did not increase in spite of high atrial pressures. Plasma ANP levels in TAH animals are almost normal. In TAH animals no role in the pathogenesis of hemodynamic abnormalities is played by abnormality of plasma ANP levels. However, TAH animals may display an abnormality in sensitivity to the p-ANP. PMID- 2973792 TI - Altered humoral control of circulating volume during artificial circulation. AB - Plasma atrial natriuretic polypeptide (ANP) levels and plasma renin activity (PRA) were measured in six left ventricular assist device (LVAD) goats, four single artificial heart (SAH) goats, i.e., LVAD animals with fibrillated hearts, and 3 total artificial heart (TAH) calves. During the 4 week experiment, the ANP levels in the SAH goats increased to 516 pg/ml from the control value of 61.8 pg/ml because of an elevated CVP of 13-21 mmHg, while those of the LVAD goats were near the lower limit of the control values. The TAH calves had slightly decreased ANP levels at 23-46 pg/ml after the third postoperative week, which did not increase after the steep increase in CVP induced by the changes in the driving conditions. The PRA levels were 10-50 times higher than control in the SAH goats and a TAH calf whose CVP rose to 19 mmHg, although the cardiac output was within normal limits. IN CONCLUSION: 1) The secretion of ANP secondary to alteration in CVP was preserved in the LVAD recipients and enhanced diuresis, while it was compromised in the TAH cases. 2) The PRA levels in cases with elevated CVPs were high enough to suppress diuresis. PMID- 2973794 TI - A double-blind clinical trial to determine the importance of pumice prophylaxis on fissure sealant retention. PMID- 2973795 TI - Interparietal hernias: analysis of six cases with literature review. PMID- 2973793 TI - Pulmonary emboli from blood-biomaterial interaction. AB - The problem of surface thrombosis and subsequent embolization remains entrenched as a yet incompletely surmounted barrier to the development of truly satisfactory intravascular prosthetic devices. A baboon ex vivo shunt was used to determine the interaction of Indium-111 platelets and potential biomaterials. The uptake of Indium-111 platelets was monitored continuously by gamma camera scanning. Several of the materials tested demonstrated a saw-toothed pattern of platelet activity, with accumulation followed by rapid decline. Neither PTFE nor Dacron exhibited this pattern. Post shunt scans of the animals' chests showed discrete foci of platelet activity in the lungs, corresponding to each embolic event noted on the material's scan. In conclusion, the search for a smooth surface as a blood material interface may produce a material which accumulates and then sloughs significant platelet aggregates. It is crucial that these materials be subjected to vigorous testing to determine their safety prior to initiation of clinical trials. PMID- 2973796 TI - A five- to seven-year experience with externally-supported Dacron prostheses in axillofemoral and femoropopliteal bypass. AB - We have examined the clinical results of 56 externally-supported (EXS) Dacron grafts in the axillofemoral position and 117 in the femoropopliteal position. Results have been analyzed from two perspectives: primary patency concerns only those grafts that had never occluded; extended patency refers to all open grafts including those whose continued patency is the result of thrombectomy. The 5-to 7 year life-table patency rates are: axillofemoral 8 mm and 6 mm bypass: primary 75% and extended 97%; above-knee femoropopliteal 6 mm bypass: primary 78% and extended 93%; below-knee 6 mm femoropopliteal bypass: primary 41% and extended 91%. In contrast, the results for the 5 mm grafts used for femoropopliteal bypass were inferior to the 6 mm grafts: femoropopliteal 5 mm bypasses had an above-knee primary patency rate of 44% and an extended rate of 55%, with a below-knee primary patency rate of 15% and an extended rate of 32%. Rendering the noncrimped porous Dacron prosthesis kink and compression resistant by an external support coil appears to increase its potential for successful use, especially in 8 mm axillofemoral and 6 mm femoropopliteal bypasses. PMID- 2973797 TI - Long-range observations following two-stage occlusion of the descending thoracic aorta for type III dissection. AB - Clinical and angiographic observations have been presented in a patient with subacute dissecting aneurysms of the descending thoracic aorta (Type III) who underwent a two-stage operation. The first stage consisted of insertion of an ascending to lower abdominal aorta bypass graft through a midline incision, the second stage of exclusion of the dissected segment through a separate left thoracotomy a week later. The patient remained asymptomatic through the two years' observation period. Repeated angiograms showed persistent obliteration of the dissected segment and good flow through the aorto-aortic bypass. PMID- 2973798 TI - Anastomotic false aneurysms with aortic Dacron graft after twenty-five years. AB - One of the early diamond crimped knitted polyester (Dacron) grafts was surgically excised after implantation for 25 years in the aorto-biiliac position because of false aneurysm formation at the three anastomotic sites. The sutures were no longer visible. While the areas around the false aneurysm were poorly incorporated, the graft limbs were well encapsulated with some endothelial-like cells on the luminal surface. The integrity of the graft was well preserved despite mild fraying and the disruption of one stitch. PMID- 2973799 TI - Mycotic suprarenal aortic aneurysm. AB - Two cases of mycotic suprarenal aortic aneurysm treated by anatomic reconstruction are presented. Diagnosis was facilitated by 67-Gallium Citrate Scanning, Computer Assisted Tomography (CT), and Angiography. Increasing symptoms with attenuation of the aortic wall identified by CT necessitated urgent resection and reconstruction in both patients. Contained rupture of the aneurysm was found in one patient. Cultures of the resected tissue grew Salmonella dublin and Bacteroides fragilis. The patients were treated with continuous antibiotic therapy. Follow-up demonstrated asymptomatic patients with sterile blood cultured and good anatomic results on CT at one year. No recurrent sepsis nor aneurysmal dilatation is evident to date. Based on our experience and the available literature we conclude that inline reconstruction is the method of choice for treatment on these lesions. PMID- 2973800 TI - Combined aortobifemoral and femoropopliteal bypass in the management of patients with extensive atherosclerosis. AB - Certain patients have atherosclerosis in both aortoiliac and femoropopliteal segments of the arterial tree and thus do not have a good result from reconstruction of the aortoiliac segment. No method has been developed to identify these patients and we do not know whether, by combining a femoropopliteal bypass with an aortobifemoral bypass, the results can be improved. We present a series of 153 patients with severe multilevel occlusive disease treated by simultaneous reconstruction and followed for up to 6.5 years. The cumulative patency of the femoropopliteal bypasses was 80% at four years. Functional and symptomatic improvement was excellent, and operative mortality was low when one considers the age and poor general condition of the patients. PMID- 2973801 TI - Interposition of a prosthetic patch to prevent aorto-enteric fistula. AB - Aortoenteric fistula is an uncommon but disastrous complication of aortic reconstruction with prosthetic grafts. Therefore, prevention of enteric erosion when using a vascular prosthesis must be a primary concern. A flap of greater omentum can provide an adequate coverage of the graft when retroperitoneal tissue is insufficient. If the omentum is not available due to atrophy or previous abdominal surgery, interposition of a prosthetic patch represents another means of preventing duodenal erosion. Polytetrafluoroethylene is our choice of material because of its softness, elasticity, and thickness. PMID- 2973802 TI - Ruptured aortic aneurysms: postoperative complications and their management. AB - Postoperative complications in 92 patients undergoing repair of ruptured abdominal aortic aneurysms are reviewed. Renal failure and cardiac complications were fatal in 74% and 85% of the patients, respectively. Preoperative renal function at the time of presentation had no correlation with the development of renal failure. All but one patient in whom the left renal vein was divided developed renal failure postoperatively. There also was a preponderance of this complication in the patients needing suprarenal aortic control. Respiratory complications were seen in 29% of patients with 22% mortality. Peripheral emboli and ischemic colitis developed in 8% and 9% of the patients, respectively. PMID- 2973803 TI - Effect of healing on compliance of porous Dacron grafts. AB - The effects of tissue ingrowth on the compliance of porous Dacron grafts were studied in 8 mm external-velour warp-knit Dacron grafts implanted in the canine descending thoracic aorta. The compliance of these grafts was measured before implantation and after implant periods of two weeks and one, two, four, and six months. Compliance, which initially was 0.043 +/- 0.008% mm Hg, decreased less than 10% after implantation (no statistical difference at p less than 0.05). No anastomotic neointimal hyperplasia was observed. Although the total thickness of the graft wall increased two- to three-fold after implantation, the benign healing process observed in this study had very little influence on graft compliance followed to six months. PMID- 2973804 TI - Evaluation and preparation for surgical treatment of renal artery disease. AB - The diagnosis of renovascular disease and renovascular hypertension is outlined. A comparison and analysis of the advantages of three forms of treatment are made. These include medical management, percutaneous transluminal coronary angioplasty, and surgical intervention. Selection of patients for revascularization to preserve renal function is discussed, and guidelines for determining renal salvageability are presented. Surgical revascularization is the treatment of choice for patients with ostial atherosclerotic renal artery disease, branch renal artery disease, or a renal artery aneurysm. PMID- 2973805 TI - Ca2+-dependent phospholipid- (and membrane-) binding proteins. PMID- 2973806 TI - Free fatty acid enhancement of cation-induced fusion of liposomes: synergism with synexin and other promoters of vesicle aggregation. AB - The effect of free fatty acids on the cation-induced fusion of large unilamellar vesicles (liposomes) was investigated by using fluorescent assays which monitor the mixing of aqueous contents of liposomes. Overall fusion was modeled as a two step process involving aggregation of vesicles followed by actual fusion. Different experimental conditions were used which favored either aggregation or fusion as the rate-limiting step in the overall process. When phosphatidylserine liposomes were induced to fuse by 4 mM Ca2+ plus 5 mM Mg2+, preincubation with arachidonic acid showed a dramatically increased overall rate of fusion compared to the same liposomes not treated with fatty acid. When fusion was induced by 3 mM Ca2+, arachidonic acid had little effect. These results were interpreted in terms of the action of arachidonic acid only at the fusion step per se and not the aggregation step. Therefore, the enhancement of the overall fusion rate would be observed solely under conditions where the actual fusion of liposomes was rate limiting (Ca/Mg) rather than the aggregation of liposomes (Ca alone). When other liposome systems were tested, the effect of arachidonic acid was observed only under fusion rate-limiting conditions. Arachidonic acid was found to act synergistically with promoters of liposomal aggregation, such as Mg2+, spermine, and synexin, to enhance the overall rate of liposome fusion, as would be expected from action at separate kinetic steps. The dependence of the fusion rates on arachidonic acid concentration demonstrated an apparently cooperative effect. The structure of the fatty acid is of critical importance in determining its effects, as shown by the fact that 16-doxylstearic acid always increased the rate of fusion while 5-doxylstearic acid always decreased the rate of fusion under all conditions tested. A number of different fatty acids, including oleic acid, elaidic acid, 16-doxylstearic acid, myristic acid, and stearic acid, were effective at increasing the fusion rate to varying extents. In general, unsaturated fatty acids were more effective than saturated ones, either due to partitioning into the membrane or because of structural requirements for promotion of fusion. PMID- 2973807 TI - Effect of phospholipid:protein ratio on the state of aggregation of the (Ca2+ Mg2+)-ATPase. AB - The organization of the (Ca2+-Mg2+)-ATPase has been studied in reconstituted systems by fluorescence polarization of the ATPase labeled with fluorescein isothiocyanate (FITC) and resonance energy transfer between ATPase labeled with FITC and with eosin isothiocyanate (EITC). The fluorescence polarization of FITC ATPase was found to decrease with increasing labeling ratio FITC:ATPase, indicating depolarization as a result of resonance energy transfer between ATPase molecules. Fluorescence polarization was, however, independent of the molar ratio of phospholipid to protein above a molar ratio of 50:1. Resonance energy transfer between FITC-ATPase and EITC-ATPase was also found to be independent of phospholipid:protein ratio. It is suggested therefore that the ATPase is not randomly distributed in the plane of the membrane but rather forms ordered clusters (probably rows of monomers or dimers) on the fluorescence time scale (nanoseconds) even in the presence of a large excess of phospholipid. This organization within the membrane is dependent both on the chemical structure of the phospholipid and on its physical phase. PMID- 2973808 TI - Formation of D loops by the UvsX protein of T4 bacteriophage: a comparison of the reaction catalyzed in the presence or absence of gene 32 protein. AB - The UvsX protein of T4 bacteriophage will catalyze the formation of D loops between linear single-stranded DNA (ssDNA) and homologous supercoiled double stranded DNA (dsDNA) in the absence of T4 gene 32 protein (gp32). This reaction requires one monomer of UvsX protein per three nucleotides of ssDNA so that the ssDNA is completely covered with UvsX protein. Under these conditions, high rates of ATP hydrolysis are observed, and one-third of the products are joined paranemically. The reaction proceeds through a mechanism that creates homology independent coaggregates of UvsX protein, dsDNA, and ssDNA. When UvsX protein is added to only 1 monomer per 8 nucleotides, but with 1 monomer of gp32 per 12 nucleotides, the rate of ATP hydrolysis is depressed, but D-loop formation is enhanced. Nearly all of the product is bound in plectonemic joints, and no coaggregated intermediates are formed. Coaggregate formation at high concentrations of UvsX protein is not inhibited by the presence of gp32; gp32 simply allows for efficient formation of D loops at such low concentrations of UvsX protein that coaggregates are not constructed. Electron microscopic visualization of the joint structures in this reaction reveals that both gp32 and UvsX protein are bound to the ssDNA. The single-stranded DNA binding (SSB) protein of Escherichia coli will substitute only partially for gp32: in the presence of SSB protein, D-loop formation can be catalyzed at one UvsX protein monomer per eight nucleotides, and it is accomplished without the formation of coaggregates, but a major portion of the product is joined paranemically. PMID- 2973809 TI - Preparation and characterization of heavy meromyosin and subfragment 1 from vertebrate cytoplasmic myosins. AB - The soluble fragments of myosin, heavy meromyosin (HMM), and subfragment 1 (S-1) have been instrumental in elucidating the kinetic mechanisms of the actin activated MgATPase activity of both skeletal and smooth muscle myosin. To date, relatively little has been published on these fragments from vertebrate cytoplasmic myosins. We now describe the preparation and steady-state kinetic characterization of S-1 and HMM from human platelet and avian intestinal epithelial brush border myosin. The HMM prepared from each of these tissues was similar both in their SDS-polyacrylamide gel pattern and in their steady-state kinetic properties. The Vmax of the actin-activated MgATPase activity varied between 0.8 and 2.5 s-1, and the KATPase (the apparent dissociation constant derived from a double-reciprocal plot of the MgATPase activity) was about 1-2 microM. This low value for the apparent dissociation constant was similar to the dissociation constant of HMM for actin directly measured under similar conditions and is about 40 times lower than that determined with avian smooth muscle HMM. The KATPase of the cytoplasmic HMM was only slightly increased when the ionic strength was raised from 12 to 112 mM. PMID- 2973810 TI - Cooperative turning on of myosin subfragment 1 adenosinetriphosphatase activity by the troponin-tropomyosin-actin complex. AB - In the field of muscle regulation, there is still controversy as to whether Ca2+, alone, is able to shift muscle from the relaxed to the fully active state or whether cross-bridge binding also contributes to turning on muscle contraction. Our previous studies on the binding of myosin subfragment 1 (S-1) to the troponin tropomyosin-actin complex (regulated actin) in the absence of ATP suggested that, even in Ca2+, the binding of rigor cross-bridges is necessary to turn on regulated actin fully. In the present study, we demonstrate that this is also the case for the turning on of the acto.S-1 ATPase activity. By itself, Ca2+ does not fully turn on the acto.S-1 ATPase activity; at low actin concentration, there is almost a 10-fold increase in ATPase activity when the regulated actin is fully turned on by the binding of rigor cross-bridges in the presence of Ca2+. This large increase in ATPase activity does not occur because the binding of S-1.ATP to actin is increased; the binding of S-1.ATP is almost the same to maximally turned-off and maximally turned-on regulated actin. The increase in ATPase activity occurs because of a marked increase in the rate of Pi release so that when the regulated actin is fully turned on, Pi release becomes so rapid that the rate-limiting step precedes the Pi release step. These results suggest that, while Ca2+, alone, does not fully turn on the regulated actin filament in solution, the binding of rigor cross-bridges can turn it on fully. If force producing cross-bridges play the same role in vivo as rigor cross-bridges in vitro, there may be a synergistic effect of Ca2+ and cross-bridge binding in turning on muscle contraction which could greatly sharpen the response of the muscle fiber to Ca2+. PMID- 2973811 TI - NMR study of the solution conformation of rat atrial natriuretic factor 7-23 in sodium dodecyl sulfate micelles. AB - The conformation of the cyclic portion (7-23) of naturally occurring rat atrial natriuretic factor, ANF(1-28), has been examined in sodium dodecyl sulfate (SDS) micelles using high-resolution NMR techniques. Evidence is presented which shows that ANF(7-23) has several regions of definable structure in SDS micelles which were not observed in earlier studies in bulk solvents. The 1H NMR resonances of ANF(7-23) in SDS micelles were assigned using sequential assignment techniques, and the conformational properties were analyzed primarily from proton-proton distances obtained from the quantitative analysis of two-dimensional nuclear Overhauser effect spectra. Three-dimensional structures consistent with the NMR data were generated by using distance geometry and constrained minimization/dynamics. Several similar but not identical structures were found which adequately satisfied the NMR constraints. Although none of the structures adopted a standard secondary structure, the conformations of three different sections of the peptide, 8-13, 14-17, and 18-21, were nearly identical in all of the predicted structures when individually superimposed. PMID- 2973812 TI - Ethionine-induced alterations of enzymes involved in lipid metabolism and their possible relationship to induction of fatty liver. AB - Changes of enzymes involved in the hepatic metabolism of long-chain fatty acids (palmitoyl-CoA synthetase (EC 6.2.1.3), carnitine palmitoyltransferase (EC 6.2.1.3), glycerophosphate acyltransferase (EC 2.3.1.15)) in the liver of male rats were examined after ethionine exposure. Ethionine administration resulted in a dose- and time-dependent enhancement of the palmitoyl-CoA synthetase activity both in the mitochondrial, peroxisomal and microsomal fractions. The total carnitine palmitoyltransferase activity in the mitochondrial fraction was enhanced. Ethionine administration was also associated with dose- and time dependent changes of the microsomal glycerophosphate acyltransferase activity, whereas the mitochondrial enzyme activity was marginally affected. The hepatic triacylglycerol content of the ethionine-treated animals was increased. Hepatic lipids were accumulated in large droplets. Serum triacylglycerol and cholesterol were decreased. In particular, the serum HDL-cholesterol level was lowered. The concentration of ATP in the liver decreased. Accumulation of the metabolic product S-adenosylethionine (AdoEth) was observed for the first 2 days of exposure followed by a fall in S-adenosylmethionine (Ado-Met) during the next 10 days. Linear regression analysis of ATP content versus AdoEth and AdoMet showed highly significant correlations. A significant correlation between the hepatic triacylglycerol and AdoEth content was also observed upon ethionine treatment. The data show that ethionine perturbs the hepatic lipid metabolism. Enhanced esterification of long-chain fatty acids, but not a simple reduction of their oxidation, might contribute to ethionine-induced fatty liver in addition to a block in secretion of lipoproteins and decreased protein synthesis. PMID- 2973813 TI - Acyl-CoA ligases from rat brain microsomes: an immunochemical study. AB - Acyl-CoA ligase activities, solubilized from rat brain microsomes, were fractionated into three different peaks by hydroxyapatite chromatography. Based on physical and chemical properties, we suggested that peak A (pamitoyl-CoA ligase) and peak C (lignoceroyl-CoA ligase) were two different enzymes (A. Bhushan, R. P. Singh, and I. Singh (1986) Arch. Biochem. Biophys. 246, 374-380). We raised antibodies against purified liver microsomal palmitoyl-CoA ligase (EC 6.2.1.3) and examined the effect of this antibody on acyl-CoA ligase activities for palmitic, arachidonic and lignoceric acids in microsomal enzyme extract and different acyl-CoA ligase peaks from the hydroxyapatite column. In an enzyme activity assay system in microsomal extract, the antisera inhibited the palmitoyl CoA ligase activity but had very little effect on the acyl-CoA ligase activities for arachidonic and lignoceric acids. This antisera inhibited the acyl-CoA ligase activities for these three fatty acids in peak A and had no effect on these activities in peak B or peak C. Western blot analysis demonstrated that antibody to liver microsomal palmitoyl-CoA ligase cross-reacted with only peak A (palmitoyl-CoA ligase), but not with peak B or peak C. This immunochemical study demonstrates that palmitoyl-CoA ligase does not share immunological determinants with acyl-CoA ligases in peaks B or C, thus demonstrating that palmitoyl-CoA ligase (peak A) is different from the arachidonoyl-CoA and lignoceroyl-CoA ligase activities in peaks B or C. PMID- 2973814 TI - Tissue distribution of [3H]cholesteryl linoleyl ether-labeled human Lp(a) in different rat organs. AB - The sites of tissue uptake of human lipoprotein(a) (Lp(a] were studied in rats using [3H]cholesteryl linoleyl ether [( 3H]CLE) as a marker. Since rat plasma has no cholesteryl ester transfer activity, the amount of label in various tissues should reflect the quantitative uptake of Lp(a). Isolated Lp(a) was labeled with [3H]CLE by incubation overnight of Lp(a), a source of cholesteryl ester transfer activity (1.23 g/ml infranate of human plasma), and [3H]CLE-labeled Intralipid. Following labeling, the homogeneity and integrity of Lp(a) was shown by agarose electrophoresis and immunoblotting. Intact Lp(a) was injected via the tail vein of rats (120-170 g, n = 4 at each time point), and tissues were collected at various times thereafter (4-48 h). The disappearance curve of [3H]CLE-labeled Lp(a) from rat plasma was bimodal and had an initial rapid t1/2 of 1.8 h followed by a slower component, t1/2 = 13.3 h. Tissue uptake at all sampling times was greatest in liver (28.5% at 48 h of total dpm injected), followed by the intestine (9-12%), with less than 3% uptake by spleen. The small intestine was divided into four segments, and while the 3H radioactivity was similar in the proximal segments, a time-related increase in [3H]CLE was seen in its most distal portion. These studies indicate that the tissue sites of degradation in the rat of human Lp(a) are similar to human low-density lipoproteins (LDL); the increase in label in the distal portion of the small intestine with time may represent [3H]CLE excreted through the bile and absorbed by the mucosal cells. PMID- 2973815 TI - Viscoelasticity of fibrinogen solution and of blood during coagulation studied by a new damped oscillation rheometer. AB - The behavior of a newly developed damped oscillation type rheometer was analyzed for fibrinogen solution and blood during coagulation. This rheometer consists of a cylindrical tube suspended from a torsion wire, that is filled with liquid to be tested. The logarithmic damping factor (LDF) during coagulation for blood and fibrinogen solution was obtained by this rheometer, which was closely related to the changes of viscosity and/or viscoelasticity of the blood sample. The slight increase of LDF prior to the rapid decrease was observed for blood. The increase of LDF would be reflected in the formation of the aggregation structure of red blood cells (rouleaux network) prior to the formation of fibrin network. The value of LDF for fibrinogen solution sharply increased and then decreased through a maximum value with the progress of coagulation, although the change of LDF was remarkably dependent on the fibrinogen concentration. The initial increase in LDF for fibrinogen solution was considered to be due to the formation of small clots in the solution. The decrease in LDF after attaining a maximum value is ascribed to the formation of fully developed fibrin network. The maximum value of LDF during coagulation for fibrinogen solution is higher than that for blood. The behavior was compared with that for non-biological fluids such as viscosity standard liquids and polyvinyl alcohol solution. From those data, it was concluded that the higher value of LDF than that for Newtonian liquids was due to the formation of aggregation structure or inhomogeneous fine clots in the liquid, which was accompanied with the appearance of the elasticity. PMID- 2973816 TI - Trial of the RBC aggregometer head for estimating blood flow in veins in vivo. AB - The whole blood RBC aggregometer head reported previously for measuring the degree of RBC aggregation in whole blood was tested for its usefulness as a flowmeter of blood vessels in situ. Modifications to its construction were made so that it became readily attachable and detachable without damage to the vessels. In ex vivo experiments employing a transparent vinyl tube and freshly drawn heparinized human whole blood, the RBC aggregometer head was applicable for evaluating semiquantitative flow changes within a limited flow range based on the degree of RBC aggregation. A linear relationship was observed between the logarithm of blood flow in a low shear range (below approximately 180/s) and changes in the light transmission of the flowing blood. The RBC aggregometer head with or without an electromagnetic flowmeter (EMF) was applied to the jugular vein and femoral vein in cats. A stop-flow change of whole blood in the jugular vein was detected by the RBC aggregometer head as a dramatic change in light transmission (LT). The aggregometer head recorded a similar LT change consistently, whereas the EMF was found to be rather discrepant, indicating the occurrence of anomalous flow. It is concluded that the RBC aggregometer head can be used as an semiquantitative flowmeter for relative changes in blood flow in veins in situ. PMID- 2973817 TI - [Quantitative assessment of the blood supply to the focus of myocardial ischemia in dogs in pharmacological research]. AB - The method of quantitative evaluation of blood supply of ischemic myocardial focus was suggested, it consists in measuring the venous blood outflow from the ischemic area with the help of ultrasonic technique. In experiments on anaesthetized dogs with coronary artery occlusion the rate of blood flow and dynamics of its changes in the ischemic focus were determined. It was shown that sodium oxybutyrate improves the blood supply of myocardial ischemic area. PMID- 2973818 TI - [The action of tuftsin on the reaction of macrophage-suppressor formation in vitro and in vivo]. AB - A cell suspension consisting of nonadhering and adhering spleen cells in the ratio 30:1 was incubated in a 10(-4) M tuftsin's solution during 15-30 min. The addition of 10(7) cells incubated in tuftsin syngeneic recipients resulted in the suppression of the immune response of the latter to sheep red blood cells. It was noted that this effect may be induced by using adhering cells only of intact donors and only when incubated together with nonadhering cells. The addition of tuftsin one hour after transplantation of the nonadhering spleen cells resulted in the suppression of the immune response. It was proposed, that suppression effect released through generation of the macrophage-suppressors. PMID- 2973819 TI - Lymphokine-activated killer cells from normal and lymphoma subjects are cytotoxic for cells coated with antibody derivatives displaying human Fc gamma. AB - Lymphokine-activated killer (LAK) cells were successfully generated in all cases from blood mononuclear cells obtained from six patients with lymphoma. The LAK cells from three of these patients and from five normal adult donors were tested for their effector abilities in antibody-dependent cellular cytotoxicity (ADCC) against guinea pig leukemic lymphocytes coated with various antiidiotype antibodies. Cells from all the donors behaved similarly. Mouse monoclonal antibodies of IgG1, IgG2a, and IgG2b isotypes invoked no ADCC. However, substantial ADCC was invoked by the chimeric antibody FabFc, in which Fab'gamma from mouse antiidiotype is thioether-bonded to human normal Fc gamma. Similar results were obtained on testing LAK cells from a normal donor against uncultured human lymphoma targets coated with native or chimeric antiidiotype. The ADCC invoked by the mouse-human chimeric antibodies appears to depend on the human Fc gamma they display and not on the univalency of the derivatives used. The findings imply that LAK technology could usefully augment serotherapy that uses antibody derivatives displaying human Fc gamma. PMID- 2973820 TI - Medroxyprogesterone acetate enhances spontaneous mammary tumorigenesis and uterine adenomyosis in mice. AB - Based on our previous findings that medroxyprogesterone acetate (MPA) significantly suppressed the formation of preneoplastic mammary hyperplastic alveolar nodules in mice, and that this suppression persisted for some time, we studied the effects of different schedules of MPA treatment on spontaneous mammary tumorigenesis and uterine adenomyosis in SHN virgin mice. Mice received a subcutaneous pellet of MPA every 2 months: I) during the limited period of 1-3 months of age; II) throughout the experiment beginning at 6-8 months of age; and III) throughout the experiment beginning at 2-3 months of age. All treatments significantly enhanced mammary tumorigenesis with little difference in the effects among treatments. The progression of uterine adenomyosis was also stimulated in Experiments I and III, but not in Experiment II. These results are in good accord with our previous observations with progesterone, indicating that MPA has progesterone-like effects on mammary and uterine lesions of mice. PMID- 2973822 TI - Drugs and their hazards. PMID- 2973821 TI - Predicting who will progress to AIDS. PMID- 2973823 TI - [Teratogenic effect of pyrimethamine in the rat. In vivo prevention by calcium folinate]. AB - In vivo prevention of malformations induced by pyrimethamine was studied in the rat. The administration of calcium folinate had an almost complete antiteratogenic effect on external, visceral and skeletal malformations, provided that the vitamin was administered together with the pyrimethamine and that the treatment was continued for the following two days. PMID- 2973825 TI - [Use of acetamide by Bacillus gordonae. I. Isolation and characterization of mutants able to use this compound as a source of carbon and energy]. AB - All known strains of Bacillus gordonae can acquire, by spontaneous mutation, the ability to grow at the expense of acetamide as a source of carbon, nitrogen and energy. The isolation and characters of these mutants are described. Their frequency is high (10(-3)-10(-2) per cell). PMID- 2973824 TI - [Preparation of 125I-labelled monoclonal antibodies of the insulin receptor]. AB - Three monoclonal anti-insulin receptor antibodies have been labelled with 125I according to various methods (Cloramine T, Lactoperoxidase and IODO-GEN). The effect of labelling on antibody structure and function has been characterized using the following parameters: a) specific activity obtained in four different labelling procedures, at least; b) TCA labelled antibody precipitable 90 days after labelling; c) interaction between labelled antibodies and the insulin receptor; d) ability of antibodies to inhibit insulin-stimulated receptor auto phosphorylation. Cloramine T method produced labelled antibody with constant specific activity; however, some preparations were unstable and showed reduced capacity to recognize the insulin receptor. Lactoperoxidase method produced stable antibodies; however, specific activity was highly variable and antibodies had low capacity to interact with the insulin receptor. The IODO-GEN method produced antibodies with constant specific activity, stable, high capacity to interact with the insulin receptor, and, moreover, maintaining in full the capacity to inhibit the insulin-stimulated auto-phosphorylation of the insulin receptor, since it does not induce antibody alterations which in turn affect antibody-receptor interaction biological action. PMID- 2973826 TI - [Use of acetamide by Bacillus gordonae. II. Research on acetamidase and the taxonomic value of spontaneous mutation permitting the acquisition of this enzyme]. AB - The wild strain Q1 had no acetamidase. The mutant Q1Ac synthesized an inducible acetamidase which was catabolite repressible by glucose. The mutation described is a character that has a high taxonomic value. It constitutes a new example of acquisitive evolution. PMID- 2973827 TI - [Implication of lipid peroxidation in triethyltin poisoning in the rat]. AB - Triethyltin intoxication induces, in vivo, a significant increase of malondialdehyde concentration in rat brain. After treatment with a Ginkgo biloba extract, an extract known to possess antiedematous and radical scavenging properties, the malondialdehyde level in the brain is significantly decreased. This suggests that a lipid peroxidation process is associated with cerebral oedema induced by triethyltin. PMID- 2973828 TI - [Effect of vanadate on the vacuolar ATPase of Acer pseudoplatanus]. AB - The ATPase of vacuoles isolated from Acer pseudoplatanus cells is strongly inhibited by vanadate, a specific inhibitor of plasma membrane ATPase. The degree of inhibition depends upon the ionic composition, ATP and magnesium concentrations of the reaction medium and the inhibition is reversed by EDTA which complexes vanadate. In absence of factor which may interfere with the effectiveness of inhibitor, vanadate inhibits the ATPase non-competitively. PMID- 2973829 TI - [Communication behavior in psychotic children]. AB - This study presents a descriptive approach to communication and interactions in a psychotic group children, without adult intervention. This investigation use a ethological method. The results show correlation between children's non verbal communication degree and the degree of severity in psychiatric troubles. PMID- 2973830 TI - Dental health in continuing care facilities: Province of British Columbia. PMID- 2973831 TI - Basal plasma beta-endorphin and beta-lipotropin in patients with symptomatic and asymptomatic myocardial ischemia. PMID- 2973832 TI - [Angioplasty of a coronary bifurcation: a kissing wire technic using a new-design catheter]. PMID- 2973833 TI - Effect of lower body positive pressure on blood pressure, plasma atrial natriuretic factor concentration, and sodium and water excretion in healthy volunteers and cardiac transplant recipients. AB - The effect of one hour (40 mmHg) lower body positive pressure on blood pressure, plasma atrial natriuretic factor concentration, and urinary sodium and water excretion was studied in 10 healthy volunteers and seven cardiac transplant recipients. Both groups showed a sustained rise in blood pressure and plasma atrial natriuretic factor concentration. The healthy volunteers had a diuresis during the period of lower body positive pressure and a small natriuresis in the subsequent hour. In contrast, lower body positive pressure had no significant effect on urinary sodium and water excretion in the cardiac transplant group. The data suggest that cardiac innervation is not important as a mediator of the haemodynamic response to lower body positive pressure but is necessary for the renal response. Furthermore, small physiological rises in plasma atrial natriuretic factor concentrations do not cause a brisk natriuresis as has been reported with pharmacological plasma concentrations. PMID- 2973834 TI - Quantitation of coronary venous adenosine in patients: limitations evaluated by radioimmunoassay. AB - Experimental studies have shown that adenosine is rapidly released in response to myocardial ischaemia. To evaluate whether coronary venous adenosine release is a metabolic characteristic of myocardial ischaemia in patients, adenosine concentrations were measured by a highly sensitive and specific radioimmunoassay. In three patients with normal coronary arteries and in seven with obstructive coronary artery disease coronary venous adenosine content was measured at rest and during atrial pacing. When whole blood or plasma were extracted immediately with perchloric acid the adenosine content was found to be lower than that previously reported. Recovery studies showed that the importance of time and temperature at low adenosine concentrations had been underestimated in preceding studies. In patients with coronary artery disease coronary venous adenosine concentration increased from 106.3(48.8) nmol.litre-1 to 114.9(57.0) nmol.litre-1 (NS) during pacing and was 130.4(63.3) nmol.litre-1 (NS) 2 min after pacing. Even in the presence of lactate production enhanced adenosine release was not consistently evidenced. Furthermore, venous adenosine content did not increase in five patients undergoing coronary artery occlusion during angioplasty of the left anterior descending coronary artery. The extremely short half life of coronary venous adenosine appears to preclude its use as an index of myocardial ischaemia in patients. PMID- 2973835 TI - Quantitative analyses of atrial myoendocrine cells and plasma atrial natriuretic peptides (ANP) of the rat with special reference to the twenty-four-hour variations in secretory granules and plasma ANP concentrations. AB - Subcellular structures of atrial myoendocrine cells in the rat heart and plasma concentrations of atrial natriuretic peptides (ANP) were examined at six evenly spaced time points over 24 h, using morphometric techniques and radioimmunoassay. Myofibrils and mitochondria of the cells occupied 73.3% of the cytoplasm; 2% of the cytoplasm was occupied by secretory granules, rough endoplasmic reticulum and Golgi complexes, structures characteristic of endocrine cells. Plasma ANP concentration was maximal at 08.00 h, when the individual volume of secretory granules was minimal. The numerical density of secretory granules was increased at 12.00 h. The plasma ANP concentration was minimal at 20.00 h, when the numerical density was minimal and the individual volume was maximal. The fluctuation in plasma ANP concentrations over 24 h was thus parallel to that in the numerical densities of secretory granules and inverse to that in individual volumes. These results suggest that in rats the secretory activity of atrial myoendocrine cells increases at the beginning of the resting period, whereas it decreases at the beginning of the active phase. PMID- 2973837 TI - Use of mechanical devices for distal hemoperfusion during balloon catheter coronary angioplasty. AB - Previous attempts to protect the dependent myocardium during balloon catheter coronary angioplasty in animals and humans have had generally unsatisfactory results. This paper summarizes the authors' experience in investigating commercially available mechanical pumps for distal coronary hemoperfusion during balloon angioplasty. Both roller and piston pumps can attain adequate distal perfusion without significant side effects in the majority of patients. Our goal was to suppress angina for at least 5 min to prolong balloon inflation in awake patients. Minor T-wave changes without concomitant angina pectoris can be expected when the distal coronary bed is perfused with hypothermic blood. Side branch occlusion by the inflated balloon prevents effective protection of the corresponding part of the dependent myocardium during distal hemoperfusion, which may result in persistent angina and ST-T changes uncorrected by increasing the hemoperfusion rate. Distal coronary diffuse spasm, rare and transient, was the only immediate complication of this procedure. It is suggested that intense local wall stimulation could occur with a higher flow rate (jet effect). Improved balloon catheter pressure/flow characteristics and on-line continuous mechanical pumps should soon make distal coronary hemoperfusion through balloon catheters an accepted clinical technique. PMID- 2973838 TI - Use of temporary pacemakers during coronary angioplasty: an evolving experience with ventricular fibrillation in 400 cases. AB - Because of a perceived high frequency of ventricular fibrillation when a pacemaker catheter was placed in the right ventricular apex during PTCA, we reviewed our first 400 cases for this complication. The incidence of ventricular fibrillation in the entire group was 10 of 400 cases (2.5%) compared with 17 of 3,806 (0.4%) diagnostic coronary arteriograms performed in our laboratory during the same time period (P less than .001). Ventricular fibrillation occurred more frequently during PTCA when the pacemaker catheter tip was placed in the right ventricular apex (8 of 63 cases, 12.6%) than when it was positioned in either the pulmonary artery, vena cava, or when a pacemaker catheter was not used (2 of 337 cases, 0.6%, P less than .001). Based on these data we have developed a policy of inserting temporary pacemakers prophylactically during PTCA only for patients undergoing dilation of dominant right or dominant left circumflex coronary arteries and in patients with acute myocardial infarction. PMID- 2973840 TI - Percutaneous transluminal angioplasty of a stenosis at the origin of the left internal mammary artery graft: a case report. AB - A case of percutaneous transluminal angioplasty of a high-grade stenosis at the origin of the left internal mammary graft is presented. An investigational angled balloon dilating catheter was used. PMID- 2973839 TI - Technique for guiding catheter exchange during coronary angioplasty while maintaining guidewire access across a coronary stenosis. AB - This case report describes a technique for guiding catheter exchange while angioplasty extended guidewire access is maintained across a coronary artery stenosis with a 245-cm-0.035-in.-long wire in the aorta to exchange for a more favorable guiding catheter. The indications, technique, and outcome of a case are described; it illustrates the usefulness of maintaining angioplasty guidewire access in a difficult-to-cross coronary stenosis. PMID- 2973836 TI - Immunohistochemical and ultrastructural localisation of peptide-containing nerves and myocardial cells in the human atrial appendage. AB - The innervation and myocardial cells of the human atrial appendage were investigated by means of immunocytochemical and ultrastructural techniques using both tissue sections and whole mount preparations. A dense innervation of the myocardium, blood vessels and endocardium was revealed with antisera to general neuronal (protein gene product 9.5 and synaptophysin) and Schwann cell markers (S 100). The majority of nerve fibres possessed neuropeptide Y immunoreactivity and were found associated with myocardial cells, around small arteries and arterioles at the adventitial-medial border and forming a plexus in the endocardium. Subpopulations of nerve fibres displayed immunoreactivity for vasoactive intestinal polypeptide, somatostatin, substance P and calcitonin gene-related peptide. In whole-mount preparations of endocardium, substance P and calcitonin gene-related peptide immunoreactivities were found to coexist in the same varicose nerve terminals. Ultrastructural studies revealed the presence of numerous varicose terminals associated with myocardial, vascular smooth muscle and endothelial cells. Neuropeptide Y immunoreactivity was localised to large electron-dense secretory vesicles in nerve terminals which also contained numerous small vesicles. Atrial natriuretic peptide immunoreactivity occurred exclusively in myocardial cells where it was localised to large secretory vesicles. The human atrial appendage comprises a neuroendocrine complex of peptide-containing nerves and myocardial cells producing ANP. PMID- 2973842 TI - Percutaneous transluminal angioplasty for shepherd's crook right coronary artery stenosis. AB - Angiograms from 1,043 consecutive patients undergoing right coronary artery angioplasty were reviewed to determine the incidence of "shepherd's crook" origin of the right coronary artery. Primary results, complications, and technical difficulties were compared with a control group. Fifty-one patients (4.9%) were found to have this anatomic variation. Compared with a control group, the primary success rate was lower (86% vs. 98%) and the incidence of procedural difficulties was higher (33% vs. 13%). Technical problems led to the use of more guiding catheters and more guidewires per patient than in the control group. Thus, coronary angioplasty of shepherd's crook right coronary artery imposes technical problems and is associated with less than an optimal primary success rate. PMID- 2973841 TI - Use of angioplasty guiding catheters for improved coronary artery opacification. AB - The following report describes the use of the high flow injection characteristics of a coronary angioplasty guiding catheter to improve coronary opacification in a patient with exceedingly high coronary runoff. This technique offers improved coronary visualization in special cases. PMID- 2973843 TI - Antigen presented in the local lymph node by cells from dimethylbenzanthracene treated murine epidermis activates suppressor cells. AB - Application to skin depleted of LC by treatment with the chemical carcinogen DMBA of a dose of contact sensitizer optimal for inducing contact sensitivity activates transferrable suppressor cells. Excision of solvent- or DMBA-treated skin at various times following application of the contact sensitizer DNFB indicated that the fraction of antigen which leaves the skin within the first few hours induces tolerance. An initial signal inducing unresponsiveness, observed within 1/2 hr, was overturned 3-6 hr later. A more permanent tolerogenic signal in the DMBA- but not solvent-treated lymph node resulted from an epidermal cell from DMBA-treated skin presenting antigen to suppressor cells. Therefore it is likely that suppressor cells are activated in DMBA-treated mice by an epidermal cell which migrates to the local lymph node. Local lymph node cells from DMBA treated mice also have a diminished ability to present antigen in vivo but they do not activate suppressor cells. PMID- 2973844 TI - Subpopulations of mature murine thymocytes: properties of CD4-CD8+ and CD4+CD8- thymocytes lacking the heat-stable antigen. AB - The heat-stable antigen (HSA), recognized by the monoclonal antibodies M1/69, B2A2, and J11d, is low or absent on the surface of most murine peripheral T cells but present on all but 3% of thymocytes. The CD4-CD8+ and CD4+CD8- or "single positive" thymic populations may be divided into further subgroups based on surface HSA expression. One group, CD4-CD8+ and expressing very high levels of HSA (HSA++), is an immature, T cell antigen receptor (TcR) negative, outer cortical blast cell. However, a further subdivision of CD4-CD8+ and CD4+CD8- single positives may be made, into those negative to low for HSA (HSA-) and those expressing moderate amounts of HSA (HSA+). The proportion of HSA- single positives is low in the thymus of young mice, whereas the proportion of HSA+ single positives is similar to that of the adult. Both the HSA- and the HSA+ subsets of single positive thymocytes from adult mice are CD3+ and express the normal peripheral T cell incidence of V beta 8 determinants on the TcR. On stimulation with concanavalin A in limit-dilution culture both HSA- and HSA+ subsets of single positive thymocytes give a high frequency of proliferating clones, and the clones from both HSA- and HSA+ subsets of CD4-CD8+ thymocytes are cytotoxic. Thus both HSA- and HSA+ single positive thymocytes are functionally mature. The HSA- subsets of single positive thymocytes differ from the HSA+ subsets in being slightly larger in size, in expressing higher levels of MEL-14, in binding more peanut agglutinin, and in including a proportion of cells expressing high levels of the Pgp-1 glycoprotein. It is suggested that HSA- CD4 CD8+ and HSA- CD4+CD8- thymocytes are more mature than their HSA+ counterparts, and might represent a previously activated or "memory" thymic subpopulation. PMID- 2973845 TI - Cytolytic activity of Ia-restricted T cell clones and hybridomas: evidence for a cytolytic mechanism independent of interferon-gamma, lymphotoxin, and tumor necrosis factor-alpha. AB - Ten different helper T cell (Th) hybridomas that are specific to Ia or antigen plus Ia were found to express nonspecific cytolytic activity toward the cytotoxin (CT)-resistant P815 cells upon activation with either Con A or a monoclonal anti T3 antibody (T3-mAb). In contrast to cytolytic Th1 clones which secrete high levels of interferon-gamma (IFN-gamma) and cytotoxin (CT) (lymphotoxin (LT, also known as TNF-beta) or tumor necrosis factor-alpha (TNF-alpha], these Th hybridomas produce low or undetectable levels of IFN-gamma and CT. No inhibitory activity of IFN-gamma and CT was observed in culture supernatants of activated Th hybridomas. Double-chamber experiments demonstrated that CT-sensitive L929 cells when physically separated from activated Th1 clones were killed by membrane permeable CT. Under identical experimental conditions, lysis of P815 cells did not occur. Moreover, activation of Th hybridomas directly in wells containing the CT-sensitive L929 cells failed to induce target cell lysis. This confirms that these Th hybridomas produce little CT and argues against high local concentrations of CT being responsible for Th hybridoma-mediated killing of P815 cells. Finally, a polyclonal rabbit antiserum to rTNF-alpha, which strongly and specifically inhibited CT-mediated and Th1 clone-mediated killing of L929 cells, failed to inhibit P815 lysis by activated Th1 clones and Th hybridomas. These observations establish that a cytolytic mechanism independent of IFN-gamma, LT, and TNF-alpha is responsible for lysis of CT-resistant target cells. PMID- 2973846 TI - Alteration of the intracellular localization of the La protein compared with the localization of U snRNPs. PMID- 2973847 TI - Effects of alpha-human atrial natriuretic peptide on the interrelationship of arterial pressure, aortic nerve activity, and aortic diameter. AB - A previous study has suggested that atrial natriuretic peptide (ANP) alters arterial baroreflex control of lumbar and renal sympathetic nerve activity. To explore these mechanisms, we examined the interrelationship of arterial pressure, afferent aortic nerve activity, and aortic diameter in rabbits anesthetized with alpha-chloralose before and after sinoaortic denervation and bilateral vagotomy. Arterial pressure was decreased in stepwise fashion by intravenous infusion of alpha-human ANP (alpha-hANP, 0.1-1.0 microgram/kg/min) or sodium nitroprusside (SNP, 1-5 micrograms/kg/min). Both in rabbits with intact baroreceptors and in those with baroreceptors denervated, aortic nerve activity and the aortic diameter decreased during hypotension caused by the infusion of SNP but remained unchanged during hypotension caused by the infusion of alpha-hANP. In addition, we examined the effects of alpha-hANP and SNP on the responses of the aortic diameter and aortic nerve activity to rapid changes in arterial pressure caused by intravenous phenylephrine or nitroglycerin. Changes in aortic nerve activity and the aortic diameter in response to rapid changes in arterial pressure caused by phenylephrine or nitroglycerin were not different between the infusion of alpha-hANP and SNP. These results suggest that aortic nerve activity remains unchanged despite hypotension during the infusion of alpha-hANP, because alpha hANP dilates the aorta. Since the aortic diameter increases, strain of aortic baroreceptors does not decrease. A second suggestion is that alpha-hANP does not alter aortic baroreceptor responses to changes in arterial pressure caused by phenylephrine or nitroglycerin. PMID- 2973848 TI - Subtyping of erythrocyte phosphoglucomutase-1 as a genetic marker for bone-marrow engraftment and hematopoietic chimerism after allogeneic bone-marrow transplantation in a patient with acute lymphoblastic leukemia. AB - We report an effective follow-up of the establishment of bone-marrow function after an allogeneic bone-marrow transplantation in a patient with acute lymphoblastic leukemia, by means of a suitable genetic marker, phosphoglucomutase 1 (EC 5.4.2.2) isoenzyme. A patient with acute lymphoblastic leukemia received allogeneic bone-marrow graft from a sibling who was of the same sex and blood group, HLA-identical, and mixed-lymphocyte-culture nonreactive. To monitor the bone-marrow engraftment and the type and degree of chimerism established, we used a genetic marker, the phosphoglucomutase-1 isoenzyme system, to reveal the difference between the bone-marrow host and donor. We did phosphoglucomutase-1 isoenzyme subtyping of the host's and donor's erythrocytes before transplantation, and isoenzyme phenotyping of the host's erythrocytes during a year after transplantation. Establishment of bone-marrow graft function, a period of temporary mixed chimerism with a population of both host's and donor's erythrocytes, a period of the exclusive presence of donor's erythrocytes, and the resumed appearance of host's erythrocytes after eight months, with no signs of relapse of leukemia, were all observed by analysis of phenotypes. These isoenzymes served as a significant and practical genetic marker, which could be successfully used in studies on bone-marrow transplantation. PMID- 2973850 TI - [Study of splanchno-portal hemodynamics in prognostic evaluation of the cirrhogenous development in patients at risk]. PMID- 2973849 TI - Effect of low dose infusion of atrial natriuretic peptide on renal function in man. AB - 1. The effects of the infusion of a low dose (2 pmol min-1 kg-1 for 3 h) of human atrial natriuretic peptide (hANP) were studied in seven healthy volunteers undergoing a water diuresis. Lithium clearance was used to monitor proximal tubular function. 2. hANP increased urine flow rate, sodium, calcium and magnesium excretion without significant changes in potassium and phosphate excretion, heart rate or blood pressure. 3. hANP caused a small change in fractional lithium clearance, and larger changes in distal nephron handling of sodium and water. 4. Plasma renin activity tended to decrease during the infusion of hANP, while plasma aldosterone concentration decreased during and increased after stopping the infusion of hANP. 5. The data suggest that hANP inhibits the reabsorption of sodium and water by an action on distal segments of the nephron and perhaps the proximal tubule. Inhibition of renin and aldosterone secretion may contribute to the natriuresis. PMID- 2973851 TI - [Results of diet therapy of obesity in relation to the event that triggered the onset of the disease. Considerations 10 years after the withdrawal of the treatment]. PMID- 2973852 TI - [Sulodexide treatment approach in patients with atherosclerotic vascular disease with various localizations]. PMID- 2973853 TI - [Risk of viral infection and replacement therapy in hemophilia. Considerations on the study of a family]. PMID- 2973854 TI - [Clinical results of phosphatidylserine in 40 climacteric and elderly women with psycho-organic disorders]. PMID- 2973856 TI - [The role of fish fats in the prevention of arteriosclerosis]. PMID- 2973855 TI - [Antilipemic activity of epomediol in patients with primary hyperlipidemia]. PMID- 2973857 TI - [Chronic venous insufficiency: physiopathology, clinical aspects and therapy]. PMID- 2973858 TI - [Hormones of the pituitary-thyroid axis in obese subjects on free and low-calorie diet]. PMID- 2973859 TI - [Usefulness of propranolol in the treatment of cholinergic urticaria]. PMID- 2973860 TI - [Clinical evaluation of an amoxicillin-clavulanic acid combination in respiratory tract infections]. PMID- 2973861 TI - [Clinical considerations on the efficacy and tolerability of a new sleep-inducing benzodiazepine for elderly patients: quazepam]. PMID- 2973862 TI - [Diet therapy of obese and hypertensive patients. Efficacy and modality of the hypotensive action]. PMID- 2973863 TI - [Influence of sex and age on the prevalence of gallbladder calculi in a case series of 10,000 patients examined by echography]. PMID- 2973864 TI - [Piribedil test in cephalalgic syndromes. Diagnostic and therapeutic usefulness]. PMID- 2973865 TI - [Short-term prophylaxis with sodium cefoperazone in orthopedic surgery. Preliminary data]. PMID- 2973866 TI - [Systemic lupus erythematosus during therapy with oral estrogen-progestin. Considerations apropos of a case]. PMID- 2973868 TI - [A longitudinal study on aged women with arterial hypertension]. PMID- 2973867 TI - [Diabetes mellitus and platelet half-life]. PMID- 2973869 TI - [Pseudo-rheumatoid systemic familial chondrocalcinosis]. PMID- 2973870 TI - [Role of zinc in immunoregulation and its therapeutic use in human pathology]. PMID- 2973871 TI - [Gastric diverticula: case reports]. PMID- 2973872 TI - [Percutaneous transluminal angioplasty: an underutilized therapeutic method]. PMID- 2973873 TI - [Cell membrane transport in essential arterial hypertension: the role of cations]. PMID- 2973874 TI - [New data on myocardial infarction in the young]. PMID- 2973875 TI - [Rationale and current results of loco-regional chemotherapy in hepatic metastases of colorectal carcinoma]. PMID- 2973877 TI - [Comparison of 2 administration schedules of ampicillin in respiratory infections]. PMID- 2973876 TI - [Treatment of essential systemic arterial hypertension with captopril only. Evaluation especially in aged patients]. PMID- 2973878 TI - [Current therapeutic aspects of Chlamydia trachomatis and Mycoplasma infections in gynecology]. PMID- 2973879 TI - [Considerations on infectious endocarditis in drug addicts. Description of 3 cases]. PMID- 2973880 TI - [Relations between alcoholism and work]. PMID- 2973881 TI - [Therapeutic strategy in thyroid carcinoma]. PMID- 2973882 TI - [Lipid markers of ischemic cardiopathy risk in obese women]. PMID- 2973883 TI - [Treatment of postmenopausal and senile osteoporosis with synthetic salmon calcitonin. A clinical study]. PMID- 2973884 TI - [Medical treatment of obliterating arteriopathies of the lower limbs with vasoactive drugs]. PMID- 2973885 TI - [Chronic pancreatitis: physiopathology, clinical aspects and therapy. Study of 31 cases]. PMID- 2973886 TI - [Diuretic effects of ibopamine and its action on splanchnic hemodynamics]. PMID- 2973887 TI - [Systemic effects of drugs administered by conjunctival route]. PMID- 2973888 TI - [Cardiovascular therapy in pregnancy]. PMID- 2973889 TI - [Pathology and prevention of cadmium poisoning]. PMID- 2973890 TI - [Notes on AIDS]. PMID- 2973891 TI - [Clinical study of a new macrolide, miocamycin, in 150 patients with otorhinolaryngologic disorders]. PMID- 2973892 TI - [Effects of bezafibrate on lipoprotein lipids and apolipoproteins B and A1 in patients with primary hypercholesterolemia]. PMID- 2973893 TI - [New therapeutic strategies in influenza infections in patients at risk. Controlled study of ribavirin]. PMID- 2973894 TI - [Gastrin response to protein meal in hyperthyroid patients]. PMID- 2973895 TI - [Steroid therapy in bronchial asthma]. PMID- 2973896 TI - [Aplastic anemia induced by drugs and industrial chemical agents]. PMID- 2973897 TI - [Problems of gastrointestinal therapy in pregnancy]. PMID- 2973899 TI - [Complications, errors and accidents in performing a block of the sympathetic system]. PMID- 2973898 TI - [Our experience in the field of therapy of infections in odontostomatology with bacampicillin hydrochloride]. PMID- 2973901 TI - [Evaluation of hemorrheological drugs in microcirculatory ischemia]. PMID- 2973902 TI - [Drug-induced agranulocytosis]. PMID- 2973900 TI - [Childbirth in the older woman. Clinico-statistical assessment]. PMID- 2973903 TI - [Pharmacokinetic problems in pregnancy]. PMID- 2973905 TI - [Use of ticlopidine in a group of patients with obliterative arteriopathy of the lower extremities]. PMID- 2973904 TI - [On-off phenomena in Parkinson's disease. Continuous dopaminergic stimulation]. PMID- 2973906 TI - [Pharmacological cardioversion of paroxysmal atrial fibrillation: 3 methods compared]. PMID- 2973908 TI - [Combined cyclosporin A-fluocortolone in the therapy of ulcerative rectocolitis. An evaluation of the beneficial effects observed in a case complicated by severe immune hemolytic anemia]. PMID- 2973907 TI - [Spironolactone administered topically in the treatment of acne]. PMID- 2973910 TI - [Low-dose heparin in the prevention of venous thromboses]. PMID- 2973909 TI - [Allergy to trimethoprim-sulfamethoxazole in normal, atopic and immunodepressed subjects]. PMID- 2973911 TI - [Evaluation of the therapeutic effect of a phlebotropic drug with a new noninvasive method: rheography by reflected light]. PMID- 2973912 TI - [Chemical disinfectants in hospital practice]. PMID- 2973913 TI - [Effect of treatment with alkaline bicarbonate mineral water on the metabolic response to physical exercise in insulin-dependent diabetics]. PMID- 2973914 TI - [I.v. propafenone in supraventricular paroxysmal tachyarrhythmias in patients with Wolff-Parkinson-White syndrome. 2 clinical cases]. PMID- 2973915 TI - [Occipital epilepsy: a non-benign pediatric syndrome]. PMID- 2973916 TI - [Diffusion of hepatitis B markers in a Ferrara population]. PMID- 2973917 TI - [Polyunsaturated phosphatidylcholine combined with vitamin B complex in the treatment of chronic active hepatitis with cirrhosis]. PMID- 2973919 TI - [Diagnostic imaging of lung metastases]. PMID- 2973918 TI - [Diagnostic imaging of bone metastases]. PMID- 2973920 TI - [Determination of netilmicin in continuous ambulatory peritoneal dialysis (CAPD): comparison of 3 methods]. PMID- 2973921 TI - [Use of aztreonam in the treatment of acute and flare-ups of chronic infectious bronchopneumopathies]. PMID- 2973922 TI - [Comparative evaluation of the effects of xipamide, indapamide and hydrochlorothiazide on water-electrolyte balance]. PMID- 2973923 TI - [Epidemiology and prevention of cerebrospinal meningitis in Italy in the last 10 years]. PMID- 2973924 TI - [Endorphins between psychosomatics and psychiatric therapy]. PMID- 2973925 TI - [Problems of iron therapy in pregnancy]. PMID- 2973927 TI - [AIDS in Italy]. PMID- 2973926 TI - [Diagnostic imaging of neck masses]. PMID- 2973929 TI - [Tissue concentration of aztreonam in organs of the urogenital system]. PMID- 2973928 TI - [Addiction to cocaine and recent psychostimulant molecules]. PMID- 2973930 TI - [Examining water-electrolyte balance in liver cirrhosis with a view to therapeutic management]. PMID- 2973931 TI - [Miocamycin in the therapy of bacterial infections in childhood]. PMID- 2973932 TI - [An open study of a new formulation of the levodopa and benserazide combination in the treatment of fluctuations of motor performance in patients with Parkinson's disease]. PMID- 2973933 TI - [Pathological aging of the brain. Personal observations on the use of citicoline]. PMID- 2973934 TI - [A case of sarcoidosis treated with deflazacort, a recently synthesized glucocorticoid]. PMID- 2973935 TI - [Carcinoma of the breast in men]. PMID- 2973936 TI - [Problems of insulin therapy in pregnancy]. PMID- 2973937 TI - [Cortisone: lights and shadows]. PMID- 2973938 TI - [Comparative study of oral absorption of beta-acetyldigoxin and digoxin]. PMID- 2973939 TI - [Capillaroscopic evaluation of the effects of nifedipine in patients with Raynaud's phenomenon]. PMID- 2973940 TI - [An Italian multicenter study on the efficacy and tolerability of aztreonam in the treatment of urinary tract infections]. PMID- 2973941 TI - [Acute poisoning with atropine-like alkaloids of Datura Metel]. PMID- 2973942 TI - [Observations on the clinical use of a topical amphotericin B + halcinonide combination]. PMID- 2973943 TI - [Current aspects of the medical treatment in pelvic inflammatory disease]. PMID- 2973944 TI - [Problems of calcium therapy in pregnancy]. PMID- 2973945 TI - [Non-cardiac thoracic pain. When it can be considered of esophageal origin]. PMID- 2973946 TI - [Emergencies in allergology]. PMID- 2973947 TI - [Low-sodium diet and arterial hypertension: favorable effect on HDL cholesterol of a new low-sodium polysaline combination]. PMID- 2973948 TI - [Comparative study of penbutolol and atenolol in the therapy of essential hypertension]. PMID- 2973949 TI - [Treatment with mesoglycan of chronic obliterative atherosclerotic arteriopathies of the legs. Hematochemical and flowmetric changes]. PMID- 2973950 TI - [Clinical and instrumental study of patients with syndrome X associated or unassociated with excessive myocardial oxygen consumption low work loads]. PMID- 2973951 TI - [Myocardial infarct. Contributions on the organization of work at a coronary intensive care unit in the light of current knowledge]. PMID- 2973952 TI - [Current aspects of preventive antibiotic therapy in gynecologic and obstetrical surgery]. PMID- 2973953 TI - [Diet in arterial hypertension]. PMID- 2973954 TI - [Osteosynthesis in lateral fractures of the neck of the femur]. PMID- 2973955 TI - [Electrophysiological and clinical effects of disopyramide in atrial arrhythmic disease in the initial stage]. PMID- 2973956 TI - [Adhesive polymers of medico-surgical importance]. PMID- 2973958 TI - [Mesoglycan in the treatment of chronic obstructive arteriopathies of the legs]. PMID- 2973957 TI - [Chronic obstructive bronchopneumopathy in respiratory pathology in Taranto]. PMID- 2973959 TI - [Upper digestive hemorrhage of iatrogenic cause]. PMID- 2973960 TI - [Extracardiac causes of arrhythmia]. PMID- 2973961 TI - [Problems of non-steroid anti-inflammatory therapy in pregnancy]. PMID- 2973962 TI - [Active drugs in diseases of the respiratory system]. PMID- 2973963 TI - [Use of defibrotide in the treatment of Raynaud's phenomenon associated with progressive systemic sclerosis or essential mixed cryoglobulinemia]. PMID- 2973964 TI - [Use of bacampicillin in the treatment of infections at diverse localizations]. PMID- 2973965 TI - [Immunity and influenza virus]. PMID- 2973966 TI - [Magnetic resonance in the evaluation of breast pathology]. PMID- 2973967 TI - Calcium transport and catabolism of adenosine triphosphate in the protozoan parasite Giardia lamblia. AB - 1. Calcium uptake by washed trophozoites of Giardia lamblia was dependent on inorganic orthophosphate and stimulated by glucose. Uptake was both rapid and substantial: 224 +/- 73 nmoles Ca2+/mg protein/min. 2. Known inhibitors of Ca2+ uptake in mammalian cells also impeded Ca2+ influx into G. lamblia. 3. The inhibitor studies indicated that Ca2+ transport in G. lamblia was an active process. Energy for such a process could be provided by the action of ATPases. 4. Two types of ATPases were found in the parasite; one, a membrane-associated enzyme activated by Ca2+; the other, a soluble, cytosolic enzyme activated by Mg2+. 5. These enzymes differed not only in their intracellular distribution and divalent cation requirements, but also in their sensitivity to calmodulin antagonists. The particulate enzyme was sensitive to these inhibitors whereas the soluble ATPase was not. 6. Our data indicate that Ca2+ transport in G. lamblia is mediated by a membrane-bound, calmodulin-regulated, Ca2+-ATPase. PMID- 2973968 TI - Myosin isoenzymes of fish hearts. AB - 1. Myosin extracted from ventricular and atrial muscles of some fish species were analysed by native and SDS gel electrophoresis. 2. Within the single heart, distinct types of native myosin were present in ventricular and atrial tissues. 3. Ventricular and atrial isomyosins contained two classes of light chain subunits. 4. The present results support the suggestion that the presence of multiple molecular forms of myosin in the heart is a common property of all vertebrates. PMID- 2973969 TI - Lipomatous hypertrophy of the interatrial septum. Diagnosis by magnetic resonance imaging. AB - A surgically proved case of lipomatous hypertrophy of the interatrial septum is demonstrated on magnetic resonance imaging. The intensity on T1-weighted images and characteristic distribution of the lesion may make the diagnosis possible without surgical intervention. PMID- 2973970 TI - Allergic contact urticaria of the hands due to seafood in food handlers. PMID- 2973971 TI - Pseudotumor cerebri caused by isotretinoin. AB - A 16-year-old girl treated with isotretinoin at a dosage of 0.7 mg per kg each day experienced severe headaches and impaired night vision two months after the start of therapy. Bilateral papilledema and narrowing of the lateral ventricles of the brain were found. Pseudotumor cerebri and impaired night vision abated when isotretinoin was discontinued and systemic corticosteroids (dexamethasone) were administered. PMID- 2973972 TI - Skin infection provoked by coagulase-negative Staphylococcus resembling gram negative folliculitis. AB - A superficial pustular eruption with acute onset has been reported in patients with acne vulgaris receiving systemic antibiotic treatment. In all past cases the causative micro-organism was found to be a gram-negative bacterium. This case report describes a similar clinical picture under the same circumstances, in which the bacterium incriminated was a coagulase-negative Staphylococcus. PMID- 2973973 TI - Procainamide-induced urticarial vasculitis. AB - Dermatologists are often faced with the difficulty of evaluating drug reactions in patients receiving multiple medications. Unfortunately, few drugs produce distinctive lesions; many types of medications can produce identical eruptions. One common drug-induced eruption is urticaria. We report a specific eruption due to procainamide: urticarial vasculitis. PMID- 2973974 TI - [Theory and practice of continuous laparotomy suture (abdominal wound dehiscence and incisional hernia)]. AB - The ideal method of closure of abdominal wounds should prevent disruption of the abdominal wall and the incisional hernia. It should be technically easy and to perform quickly. After an experimental and a clinical trial we recommend a continuous monofilament polypropylene suture for closure of abdominal wounds (844 cases). PMID- 2973975 TI - [Suppressor cell activity in patients with idiopathic dilated cardiomyopathy]. PMID- 2973976 TI - [Payment for reports by the assistance service]. PMID- 2973977 TI - Insulin-like growth factor I stimulates the synthesis and release of prolactin from human decidual cells. AB - Recent studies suggest a role for insulin-like growth factor I (IGF-I) in the regulation of hormone release from placental, gonadal, and pituitary tissues. To examine whether IGF-I may also regulate the release of PRL from human decidual tissue, we have investigated the effect of recombinant human IGF-I on PRL release from monolayer cultures of human decidual cells exposed to IGF-I for up to 4 days. IGF-I (10-1000 ng/ml) stimulated a sustained dose-dependent increase in PRL release (half-maximal concentration, 25 ng/ml) beginning 48 h after initial exposure, but had no effect on the intracellular PRL content. The amounts of PRL released from maximally stimulated cultures on days 3 and 4 were 168 +/- 3% (mean +/- SEM) and 258 +/- 8% of control values, respectively. IGF-I-mediated effects were inhibited by cycloheximide (3.6 microM), suggesting that the increase in PRL was the result of newly synthesized hormone. The increase in PRL release was not due to a generalized effect on protein release, since IGF-I had no effect on the release of trichloroacetic acid-precipitable [35S]methionyl proteins. Radioligand competition studies indicate that the biological actions of IGF-I are mediated through interaction with the IGF-I receptor. Binding of radiolabeled IGF-I to decidual cells in suspension was specific, saturable, and displacable by unlabeled IGF-I, with a potency nearly 10 times greater than that of insulin. Furthermore, exposure of decidual cells to a monoclonal antibody to the IGF-I receptor (alpha-IR3) completely inhibited both IGF-I-mediated PRL release and specific binding of [125I]IGF-I to decidual cells. Since the actions of IGF-I occurred at physiological concentrations, these findings strongly support a role for IGF-I in the regulation of PRL secretion by human decidua. PMID- 2973979 TI - Prophylaxis of amygdala kindling-induced epileptogenesis: comparison of a GABA uptake inhibitor and diazepam. AB - The prophylactic effect of an inhibitor of synaptosomal GABA uptake, SK&F 89976-A (N-[4,4-diphenyl-3-butenyl]-nipecotic acid), on the development of amygdala kindled seizures was studied in adult female rats. For comparative purposes, the action of diazepam was also investigated. Dosages of SK&F 89976-A and diazepam which were previously shown to be the ED50 for seizure inhibition in fully kindled rats (15 mg/kg and 2 mg/kg i.p., respectively) were administered daily 30 min prior to amygdala stimulation in naive, unkindled rats. Both drugs inhibited the evolution of full kindled seizure activity and markedly suppressed kindling associated increases in the duration of behavioral and electrographic seizures. Control rats developed fully kindled stage 5 seizures after 8.9 +/- 1.1 amygdala stimulations but drug-treated rats did not progress beyond an early stage of kindled seizures as long as the animals were treated with the drugs (22 days). Diazepam produced significant CNS depressant effects throughout the course of administration but SK&F 89976-A prevented kindling with no depressant side effects. The ability of SK&F 89976-A and diazepam to inhibit the development of full amygdala kindled seizures may be related to enhancement of central inhibitory GABAergic systems. PMID- 2973978 TI - Endocytosis of receptor-bound insulin-like growth factor II is enhanced by mannose-6-phosphate in IM9 cells. AB - The insulin-like growth factor II (IGF-II), and glycoproteins containing mannose 6-phosphate (M6P), bind to two different sites of the same receptor molecule (Morgan et al, Nature 329:301, 1987). To study the interactions between the two ligands on their common receptor in intact cells, we examined the effect of free M6P on IGF-II binding and endocytosis in the IM9 human lymphoblastoid cell line. M6P, up to a 3 mM concentration, had no effect on the binding of IGF-II to the cell surface receptor of intact IM9 cells at 4 degrees C. By contrast, when IM9 cells were incubated with 125I-IGF-II at 37 degrees C, 1 mM M6P increased cell associated radioactivity by twofold. The increase was resistant to acid wash at 4 degrees C, and therefore assumed to represent endocytosed IGF-II. Acid-washable radioactivity was no different, confirming that, in intact cells, M6P does not affect IGF-II surface binding. In addition, preincubation of cells with M6P at 37 degrees C for up to 3 hours did not change the abundance of receptor on the cell surface, as measured by a subsequent 4 degrees C binding assay. We conclude that M6P causes a shift of IGF-II-occupied receptors form the cell surface to intracellular locations without affecting surface binding of this ligand in IM9 cells. The effect could be produced by the binding of M6P itself, or by the displacement of endogenous phosphomannosylated ligands. PMID- 2973981 TI - Clinical manifestations of polycystic ovarian disease. AB - Elevated androgens and/or increased sensitivity to them is at the core of polycystic ovarian disease (PCOD). In orbit around this essential element are a wide variety of signs and symptoms that may come into alignment in unpredictable fashion and frequency. This article considers the clinical history and physical examination of the patient with PCOD, ovarian morphology, the laboratory work-up of patients with PCOD, and the role of ultrasonography, magnetic resonance imaging, and laparoscopy in the diagnosis of PCOD. PMID- 2973980 TI - GABA uptake inhibitors: relevance to antiepileptic drug research. PMID- 2973982 TI - Polycystic ovarian disease: animal models. AB - The reproductive systems of human beings and other vertebrates are grossly similar. In the ovary particularly, the biochemical and physiologic processes are identical not only in the formation of germ cells, the development of primordial follicles and their subsequent growth to Graafian follicles, and eventual ovulation but also in anatomic structure. In a noncarcinogenic human ovary, hypersecretion of androgen causes PCOD. Such hypersecretion may result from a nonpulsatile, constant elevated level of circulating LH or a disturbance in the action of neurotransmitters in the hypothalamus. In studying the pathophysiology of PCOD in humans, one must be aware of the limitations for manipulating the hypothalamic-pituitary axis. Although the rat is a polytocous rodent, the female has a regular ovarian cyclicity of 4 or 5 days, with distinct proestrus, estrus, and diestrus phases. Inasmuch as PCOD can be experimentally produced in the rat, that species is a good model for studying the pathophysiology of human PCOD. These PCOD models and their validity have been described: (1) estradiol-valerate, (2) DHA, (3) constant-light (LL), and (4) neonatally androgenized. Among these, the LL model is noninvasive and seems superior to the others for study of the pathophysiology of PCOD. The production of the polycystic ovarian condition in the rat by the injection of estrogens or androgens in neonate animals, or estradiol or DHA in adult rats, or the administration of antigonadotropins to these animals all cause a sudden appearance of the persistent estrus state by disturbing the metabolic and physiologic processes, whereas exposure of the adult rat to LL causes polycystic ovaries gradually, similar to what is seen in human idiopathic PCOD. After about 50 days of LL, the rat becomes anovulatory and the ovaries contain thickened tunica albuginea and many atretic follicles, and the tertiary follicles are considerably distended and cystic. The granulosa and theca cells appear normal histologically, although some of the stromal cells appear hypertrophic. The anatomic features consequent to polycystic ovaries resulting from LL are similar to those in human PCOD, and both rat and human PCOD ovarian cells still retain the ability to respond to FSH/LH, LHRH, and unilateral ovariectomy. In the estradiol valerate rat model, although the anatomy and physiology of the ovary resemble those of PCOD patients, the progressive degeneration of the hypothalamus and the altered response of the pituitary to LHRH make this model inappropriate for studying the hypothalamic-pituitary ovarian axis in the polycystic ovary condition.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2973983 TI - Binding of Ca2+ influences susceptibility of laminin to proteolytic digestion and interactions between domain-specific laminin fragments. AB - Ca2+ was found to influence the patterns of limit digests of laminin obtained with various neutral proteases. In the presence of Ca2+, larger fragments were obtained from the central part of laminin than in its absence. This was interpreted as being due to a stabilization of the central short-arm domains of laminin by bound Ca2+. When proteolytic fragments were tested for their ability to aggregate, only large fragments containing intact short arms were active, indicating an important role for these domains in laminin self-aggregation. PMID- 2973984 TI - Structural and functional characterization of mutants of recombinant single-chain urokinase-type plasminogen activator obtained by site-specific mutagenesis of Lys158, Ile159 and Ile160. AB - Single-chain urokinase-type plasminogen activator (scu-PA) is converted to urokinase by hydrolysis of the Lys158-Ile159 peptide bond. Site-directed mutagenesis of Lys158 to Gly or Glu yields plasmin-resistant mutants with a 10-20 fold reduced catalytic efficiency for the activation of plasminogen [Nelles et al. (1987) J. Biol. Chem. 262, 5682-5689]. In the present study, we have further evaluated the enzymatic properties of derivatives of recombinant scu-PA (rscu PA), produced by site-directed mutagenesis of Lys158, Ile159 or Ile160, in order to obtain additional information on the structure/function relations underlying the enzymatic properties of the single- and two-chain u-PA moieties. [Arg158]rscu PA (rscu-PA with Lys158 substituted with Arg) appeared to be indistinguishable from wild-type rscu-PA with respect to plasminogen-activating potential (catalytic efficiency k2/Km = 0.21 mM-1 s-1 versus 0.64 mM-1 s-1), conversion to active two-chain urokinase by plasmin (k2/Km = 0.13 microM-1 s-1 versus 0.28 microM-1 s-1), as well as its specific activity (48,000 IU/mg as compared to 60,000 IU/mg) and its fibrinolytic potential in a plasma medium (50% lysis in 2 h with 2.8 micrograms/ml versus 2.1 micrograms/ml). [Pro159]rscu-PA (Ile159 substituted with Pro) and [Gly159]rscu-PA (Ile159 converted to Gly) are virtually inactive towards plasminogen (k2/Km less than 0.004 mM-1 s-1). They are however converted to inactive two-chain derivatives by plasmin following cleavage of the Arg156-Phe157 peptide bond in [Pro159]rscu-PA and of the Lys158-Gly159 peptide bond in [Gly159]rscu-PA. [Gly158,Lys160]rscu-PA (with Lys158 converted to Gly and Ile160 to Lys) has a low catalytic efficiency towards plasminogen both as a single-chain form (k2/Km = 0.012 mM-1 s-1) and as the two-chain derivative (k2/Km = 0.13 mM-1 s-1) generated by cleavage of both the Arg156-Phe157 and/or the Lys160-Gly161 peptide bonds by plasmin. These findings suggest that the enzymatic properties of rscu-PA are critically dependent on the amino acids in position 158 (requirement for Arg or Lys) and position 159 (requirement for Ile). Conversion of the basic amino acid in position 158 results in a 10-20-fold reduction of the catalytic efficiency of the single-chain molecule but yields a fully active two chain derivative. The presence of Ile in position 159 is not only a primary determinant for the activity of the two-chain derivative, but also of the single chain precursor. Cleavage of the Arg156-Phe157 or the Lys160-Gly161 peptide bonds by plasmin yields inactive two-chain derivatives. PMID- 2973985 TI - Aerobactin production as a virulence factor: a reevaluation. AB - Iron starvation is one of the major barriers that virulent bacteria must overcome in order to proliferate in the host. Virtually all microorganisms possess high affinity iron (III) transport systems mediated by low molecular weight iron specific chelators called siderophores, the synthesis of which is activated under iron-limiting conditions. Siderophore aerobactin is frequently produced by enterobacteria which cause various types of infections in humans and animals. The status of aerobactin production as a virulence factor is evaluated both from data derived from experimental infection systems and the actual presence of this siderophore in clinical isolates. Aerobactin appears to be an important contributor to extracellular pathogenesis (mostly, that of Escherichia coli strains causing septicaemia and urinary tract infections) and to the extracellular stages of growth of intracellular pathogens like Shigella. When invasive bacteria actually enter target cells, acquisition of iron seems to occur independently of siderophore production. The feasibility of an antimicrobial therapy aimed at interfering with siderophore functioning is discussed. PMID- 2973986 TI - Age-related changes in adrenal size during the first year of life in normal newborns, infants and patients with congenital adrenal hyperplasia due to 21 hydroxylase deficiency: comparison of ultrasound and hormonal parameters. AB - An adrenal size index (ASI) was specifically designed to improve ultrasound evaluation of adrenal size in the 1st year of life. In 84 newborns and infants, ASI and plasma dehydroepiandrosterone sulphate (DHEA-S) concentrations were determined to study in vivo changes in adrenal size and cortical zonal composition. ASI was higher (P less than 0.0001) in the first 2 postnatal weeks (median 60.5 mm2) than in the rest of the first year (median 39.6 mm2) and showed a negative correlation with chronological age (r = -0.41). A positive correlation existed between DHEA-S concentrations and ASI (r = 0.29) as well as ASI relative to body weight (r = 0.57). These data and changes in gland echogenicity are consistent with histopathological observations that the early decrease in adrenal size is due to involution of the fetal zone. In four patients with congenital adrenal hyperplasia (CAH) due to 21-hydroxylase (21-OHase) deficiency, ASI and plasma concentrations of pre-defect steroid were determined. Two patients with severe salt loss had markedly elevated ASI, which returned to normal during treatment. Two patients without severe salt loss had pretreatment ASI in the upper normal range, which decreased with treatment. Our findings indicate that ASI is useful in assessing physiological changes in adrenal size and, in conjunction with DHEA-S determinations, in adrenal cortex composition. ASI may aid in the early diagnosis of the salt-losing variety of 21-OHase deficiency. ASI is an instantaneously available tool which, in addition to biochemical and clinical data, can be used to monitor treatment of 21-OHase deficiency. PMID- 2973988 TI - Absence of complement receptor type 3 and lymphocyte function antigen 1 causing deficient phagocyte and lymphocyte functions. AB - We describe a patient with delayed umbilical cord detachment, recurrent bacterial infections, and inability to form pus, despite persistent leucocytosis. Immunofluorescence studies with specific monoclonal antibodies showed a severe deficiency in the expression of alpha-chains of the receptor for the C3bi fragment of C3, complement receptor type 3, and the lymphocyte function antigen 1 molecule, found on neutrophil, monocyte and lymphocyte membranes. These membrane antigen defects were responsible for abnormalities in adhesive cell functions. Polymorphonuclear leucocytes demonstrated a markedly reduced chemiluminescence response as well as an impaired nitroblue tetrazolium test and superoxide generation to a particulate stimulus (zymosan), while the responses to a soluble stimulus (phorbol myristate acetate) were normal. In addition, random migration und chemotactic response to zymosan-activated serum were impaired. The lymphocytes demonstrated abolished natural killer cell cytotoxicity as well as abnormal humoral immunity and a lack of antibody response to pertussis and tetanus antigens. PMID- 2973987 TI - Cell-mediated cytotoxicity in Down syndrome: impairment of allogeneic mixed lymphocyte reaction, NK and NK-like activities. AB - Peripheral blood mononuclear cells (PBMC) from 16 non-institutionalized patients with Down syndrome (DS) were studied with various monoclonal antibodies and analysed for natural killer (NK), and NK-like activity. Lymphocyte proliferation and cytotoxic T-lymphocyte (CTL) cytotoxicity generated in mixed lymphocyte culture (MLC) were also evaluated in 11 DS patients. Phenotypic characterization of PBMC from DS subjects confirms our previous findings of high numbers of CD8+ lymphocytes and HNK-1+, and CD16+ cells. Lymphocyte proliferation and CTL cytotoxicity generated in MLC were low or absent in most patients. NK activity was low in almost all DS patients, while NK-like cytotoxicity generated in MLC was normal in the majority and did not correlate with NK activity from unstimulated PBMC. PMID- 2973989 TI - Frusemide pretreatment blunts the inhibition of renal tubular sodium reabsorption by ANF in man. AB - The effects of atrial natriuretic factor (ANF) 15 pmol/kg/min on renal function were studied in 7 normal male volunteers during maximal water diuresis. Subjects were studied in neutral salt balance either before, or after, seven days treatment with 40 mg oral frusemide. The post-frusemide state was associated with activation of the renin-angiotensin system (RAAS) and generally higher noradrenaline levels; this state was also associated with sodium retention, mainly due to enhanced distal nephron reabsorption. Without diuretic pretreatment ANF produced a natriuresis and diuresis associated with inhibition of both proximal and distal nephron sodium reabsorption. In contrast, after frusemide pretreatment, ANF caused an increase in water excretion (urinary flow rate) but no change in sodium excretion. In the post-diuretic condition ANF did not affect renal tubular handling of sodium. The enhanced tubular reabsorption of sodium post-frusemide, and the failure of ANF to suppress this, could be due to activation of the RAAS and SNS. PMID- 2973990 TI - Striatal grafts provide sustained protection from kainic and quinolinic acid induced damage. AB - Grafts of neonatal striatal tissue were placed into the striata of adult rats. When challenged immediately with intrastriatal injections of either kainic or quinolinic acid, excitotoxic damage was prevented. Thirty days later these same graft recipients received another injection of excitotoxin. The intrastriatal grafts continued to mitigate toxin-induced damage. It is hypothesized that the grafted cells not only survive, but that they may continue to elaborate some substance or substances that prevent excitotoxin-induced injury for at least 30 days. Previous investigations indicated that grafts of neonatal striatal tissue can protect the recipient striatum from kainic acid toxicity. In the following study it is demonstrated that such grafts also protect the striatum from quinolinic acid, an endogenous excitotoxin which induces kainate-like neuronal degeneration and has been implicated in the pathogenesis of Huntington's disease. It is postulated that the salutary effect of striatal grafting may be sufficiently long lasting to mitigate a chronic toxic insult. Such grafting may therefore represent a therapy for Huntington's disease and other neurodegenerative disorders in which an endogenous or exogenous toxin has been implicated as the pathogenetic agent. PMID- 2973992 TI - Heparan sulphate with no affinity for antithrombin III and the control of haemostasis. AB - Heparan sulphate with no affinity for antithrombin III (ATIII) was observed to cause acceleration of the factor Xa:ATIII interaction by 1100-fold (k2, 7 X 10(7) M-1.min-1) and the prothrombinase:ATIII interaction by 2900-fold (k2, 2.5 X 10(7) M-1.min-1). Although high-affinity heparan sulphate catalyzed higher acceleration and at lower concentration, in natural mixtures of the two forms the activity of the no affinity form predominated. Heparan sulphate had no significant effect on the thrombin:ATIII interaction but inhibited its potentiation by heparin (Kd 0.3 microM). From the estimated concentration of heparan sulphate on the endothelial cell surface it is proposed that the non-thrombogenic property of blood vessels is due to the acceleration of the factor Xa or prothrombinase:ATIII interaction by the greater mass of surface-bound heparan sulphate rather than by the much smaller proportion of heparin-like molecules (with high affinity for antithrombin III) which may be present. PMID- 2973991 TI - Effects of the chitin synthetase inhibitor plumbagin and its 2-demethyl derivative juglone on insect ecdysone 20-monooxygenase activity. AB - The chitin synthetase inhibitor plumbagin and its 2-demethyl derivative juglone were found to inhibit in a dose-response fashion the cytochrome P-450 dependent ecdysone 20-monooxygenase activity associated with adult female Aedes aegypti, wandering stage larvae of Drosophila melanogaster, and fat body and midgut from last instar larvae of Manduca sexta. The concentration of these naphthoquinones required to elicit a 50% inhibition of the steroid hydroxylase activity in all the insects was approximately 1 x 10(-4) M. PMID- 2973993 TI - Transcellular ionic currents studied by intracellular potential recordings in Neurospora crassa hyphae. Transfer of energy from proximal to apical cells. AB - Membrane potentials, input resistances, and electric coupling in the apical parts of N. crassa growing hyphae were recorded with the aid of intracellular microelectrodes. It was revealed that the apical cells were always depolarized by 10 to 30 mV as compared to the adjacent proximal cells. The septal pore maintained an electrical resistance of 4 to 6 M omega. The calculated values of the endogenous electrical current passing through the septal pore varied between 0.5 and 1 nA. Electrical isolation of the apical cells resulted in their depolarization from 120-150 mV to 40-60 mV, characteristics of the membrane potential value of N. crassa adult hyphae with completely blocked electrogenic pumps. A simultaneous increase in the input resistance value from 15-20 M omega to 40-80 M omega was observed. The above data can be explained assuming that H+ ATPase activity was greatly lowered in the apical cells. Thus in the intact hyphae with electrically coupled cells energy is transferred from the proximal hyphal compartments to the apical ones. PMID- 2973994 TI - A Ca2+-stimulated, Mg2+-dependent ATPase activity in subcellular fractions from Schistosoma mansoni. AB - A Ca2+-stimulated, Mg2+-dependent ATPase activity was found in subcellular fractions from Schistosoma mansoni. Its specific and relative activities were higher in the heterogeneous cuticle fraction and in the microsomal fraction. The K0.5 for ATPase activation by free Ca2+ was 0.2-0.5 microM. This is the first description of an ATPase activity stimulated by Ca2+ in the micromolar range in S. mansoni. PMID- 2973996 TI - AIDS: who do you trust? PMID- 2973995 TI - Ultrastructural studies on the boundary tissue of the seminiferous tubules of different mammals. AB - The aims of our studies were to compare the ultrastructure of the boundary tissue of seminiferous tubules of various mammals (rat, mouse, hamster, guinea pig, rabbit, ram, bull and man). Visual analysis of electron micrographs revealed the similarity of structure of all layers at investigated animals. The boundary tissue consists of 4 layers: 1) amorphous inner lamina, 2) cellular inner lamina, 3) amorphous outer lamina, 4) cellular outer lamina. The outer lamina of boundary tissue of rat, mouse and hamster revealed in histochemical reactions meshes resembling honey-combs. The wall of seminiferous canalicules of bull and ram consists of more bigger and different structure than one at the other laboratory animals. The most different structure of boundary tissue in man was observed. The capillary vessels penetrate in the myofibroblastic layer, when comparted to that found in other mammals on the surface of the wall. PMID- 2973997 TI - Death sentence. PMID- 2973998 TI - Dentists deny AIDS patient, violate discrimination law. PMID- 2973999 TI - [Cave steroid allergy!]. PMID- 2974000 TI - Characterization of a glucosyltransferase activity in liver plasma membrane: modulation by cations and lipidic effectors. AB - 1. Glucosyltransferase activity incorporating [14C]glucose from UDP-[14C]glucose onto endogenous lipidic acceptors was localized primarily in the plasma membrane of liver. 2. Incubation of plasma membrane by phosphatidyl-choline liposomes loaded with dolichyl-phosphate stimulated the enzymatic activity. 3. This enzyme required Mg2+ for maximal catalitic activity. Ca2+ could substitute Mg2+. 4. Mn2+ acted as a partial non-competitive inhibitor of the Mg2+-activated glucosyltransferase. 5. This enzyme can be modulated by neutral and acidic phospholipids; the most efficient were phosphatidyl-serine and phosphatidyl inositol. 6. The enzymatic activity was not significantly changed by cholesterol alone but it is greatly enhanced by liposomes loaded with dolichyl-phosphate and cholesterol. PMID- 2974001 TI - Long-term study of serum glycosyl-transferase levels in pregnant women and their newborn infants. AB - 1. The authors have studied changes of glucosyl-, galactosyl- and sialyl transferase activities in the sera of women during pregnancy and the early post partum period, and in neonates from birth to 1 yr of age. 2. Galactosyl- and glucosyl-transferase activities throughout pregnancy were high in comparison to control values, and increased with advancing gestation. In the early post-partum period, the activities of these enzymes decreased markedly, reaching values similar to those of control subjects. 3. There were no variations, however, in serum sialyl-transferase activity at any stage of pregnancy or the post-partum period. In newborn infants, serum glucosyl- and sialyl-transferase activities were identical to control values. 4. Galactosyl-transferase activity was high at birth, decreased progressively and returned to the normal range only after 1 yr. 5. The physiological significance of these glycosyl-transferases in the sera of pregnant women and newborn infants is discussed. PMID- 2974002 TI - Preferential formation of triiodothyronine residues in newly synthesized [14C]tyrosine-labeled thyroglobulin molecules in follicles reconstructed in a suspension culture of hog thyroid cells. AB - Early processes of thyroid hormone (T4 and T3) synthesis in thyroglobulin molecules were studied using follicles reconstructed in a primary culture of hog thyroid cells under the influence of TSH. When the reconstructed follicles were incubated with 14C-tyrosine, thyroglobulin containing the labeled tyrosine was newly synthesized and in the presence of iodide, some of the labeled tyrosine residues were iodinated and coupled to produce labeled iodothyronines, T4 and T3. Coupling efficiency, especially the efficiency of T3 production, was much higher than that obtained from the average iodoamino acid composition of mature thyroglobulin from the gland, indicating a preferential iodination of hormonogenic tyrosines and synthesis of T3. The total production of T3 was higher than T4 under the present conditions. However, free labeled T4 released into the medium was more than T3 after 16 h incubation of the labeled follicles with non labeled tyrosine, suggesting the preferential liberation of T4 from the labeled peptide and/or release from the cells. PMID- 2974003 TI - Functional characteristics of cultured mouse pancreatic islets following exposure to different streptozotocin concentrations. AB - The present study was undertaken to investigate the acute and long-term effects of streptozotocin (SZ) on pancreatic islet function and survival in vitro. Isolated mouse pancreatic islets, that had been cultured overnight, were exposed to SZ (0.55-4.4 mM) or critic acid buffer in the case of the control group. The islets were examined either immediately after SZ exposure or after one week in culture. There was a marked loss of islets treated with 2.2 and 4.4 mM SZ during the culture; however, the DNA content of the remaining islets was unaffected. The islet insulin content was reduced 7 days after treatment with 2.2 and 4.4 mM SZ. At 4.4 mM the glucagon and somatostatin content of the islet was also decreased but not to the same degree as the insulin content. SZ-induced inhibition of glucose-stimulated insulin release and (pro)insulin biosynthesis was more pronounced on day 7 as compared to day 0. A similar pattern of inhibitory action of SZ was observed on islet glucose oxidation rates. Islet ATP contents were depressed on day 7 in islets exposed 4.4 mM SZ, but were otherwise similar to the control group. Islet NAD + NADH contents were decreased by 50% after exposure to 2.2 mM SZ, compared to the control islets on day 0. This decrease in NAD + NADH contents was to a large extent restored during the one-week culture. The present study shows that islets failed to completely repair the acute damage caused by SZ, and that the impairment of the islet glucose-stimulated insulin release induced by SZ seemed to progress in culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974004 TI - [The liver of the white rat as an experimental model of observations of the effect of ethyl alcohol on the organism]. AB - 120 d after the experimental administration of ethanol to laboratory animals (mature male white rats), the changes manifesting themselves by the steatosis and the disorders of the carbohydrate balance and of the activity of the respiratory and the hydrolytic enzymes were observed in the animals' livers. There were no symptoms of a liver fibrosis. PMID- 2974006 TI - [The origin of an extra chromosome 21 in families of children with Down syndrome]. AB - These are the first studies on the origin of nondisjunction of trisomy 21 in the USSR. Parental contribution was established in 84 of 140 families observed. In 66% cases the nondisjunction took place in oogenesis and in 34% cases - in spermatogenesis. Among the children, who inherited the additional chromosome from father, boys predominate. Compilative work on all the data available concerning the origin of the 21 nondisjunction has been performed; the factors favouring nondisjunction in I and II mitotic divisions in female meiosis, both genetical and age-dependent, have been considered. The great importance of the disturbances taking place in spermatogenesis for etiology is emphasized. It is proved that somatic hyperploidy does not serve as an indicator of predisposition for chromosome nondisjunction in meiosis. PMID- 2974007 TI - [Offspring of patients with Down syndrome]. AB - An incident of birth of a child in 16-year-old patient with typical phenotype of Down's syndrome is described. The karyotype of the proband is 47, XX + 21, while that of the child is 46, XX. Analysis of the literature data and the author's observations showed that the total ratio of trisomics and non-trisomics in the offspring of women with Down's syndrome is 9:18, which considerably differs from theoretically expected and suggests that selection against anomalous gametes exists. PMID- 2974005 TI - Regulation of two nested proteins from gene 49 (recombination endonuclease VII) and of a lambda RexA-like protein of bacteriophage T4. AB - Phage T4 gene 49, encoding recombination endonuclease VII, specifies, by initiation from an AUG and an internal GUG codon, two in-frame overlapping peptides (of 18 and 12 kD). The gene is transcribed early and late, albeit from different promoters. The sequence predicts that in long early transcripts, initiated far upstream of the coding sequence, the Shine-Dalgarno sequence of the first ribosome binding site can be sequestered in a hairpin and/or cleaved. These processes might reduce initiation from the first AUG and facilitate initiation of the 12-kD peptide from the internal GUG. The potential of this hairpin to participate in Y structures or cruciforms suggests possible autoregulation. Shorter, more stable late transcripts initiated from a late promoter immediately upstream of the first ribosome binding site cannot form this hairpin. More efficient translation of the longer 18-kD gene 49 peptide from these late transcripts accounts for the strong dependence of endonuclease VII activity on late gene expression. An ORF downstream from gene 49 can be translated from a motA-dependent transcript that starts inside gene 49 as well as from the gene 49 transcripts. Its initiation codon overlaps the stop codon of gene 49, suggesting some coupling of translation. The deduced protein resembles, among others, the RexA protein of phage lambda. Possible implications for T4 recombination and for the interference of lambda lysogens with T4 gene 49 and rII mutants are discussed. PMID- 2974008 TI - Chemotherapy of experimental toxoplasmosis with special reference to robenidine. AB - Robenidine as an anticoccidial agent alone (in doses of 50 and 100 mg/kg) or in association with two other antiprotozoal compounds--pyrimethamine and sulfadoxine (in doses of 2.5 and 250 mg/kg, respectively) were tested in the treatment of chronically infected mice with an avirulent cyst-forming HF strain of Toxoplasma gondii. The efficacy of the used drugs was evaluated by the cysts number in the brains of treated mice versus control group, antibody level and viability of survived cysts by the consecutive infection of mice. According to these criteria none of the tested drugs alone or combined exhibited to be able to produce an effective cure of cyst-stage of toxoplasmosis in experimental mice. PMID- 2974009 TI - Eight weeks of streptozotocin-induced diabetes influences the effects of cold stress on immunoreactive beta-endorphin levels in female rats. AB - Cold stress produced a significant reduction in the concentration of immunoreactive beta-endorphin (IR-BE) in the anterior pituitary of diabetic female rats. IR-BE levels in the anterior pituitary of non-diabetic female rats were not affected by exposure to the cold. The effects of cold stress on IR-BE levels in the neurointermediate lobe of the pituitary and the hypothalamus were attenuated in diabetic as compared to control animals. These data suggest that in female rats, eight weeks of diabetes produced alterations in the neuroendocrine mechanisms which modulate IR-BE levels in the pituitary and hypothalamus in response to cold stress. PMID- 2974010 TI - Insulin-like growth factor I receptors on human erythrocytes from normal children: relationship with age. AB - The binding of insulin-like growth factor I (IGF I) on red blood cells has been studied in 13 children aged 8 months to 11 years and in 10 adults. The Scatchard analysis showed a curvilinear regression. In adults, the specific binding was 4.1% of the tracer, the mean number of high affinity receptor sites per cell (Ro1) being 0.88 (K1 = 10.74 nM-1) and the mean number of low affinity receptors sites (Ro2) per cell being 7.14 (K2 = 0.37 nM-1). In children the specific binding ranged from 3 to 6.5%. Ro1 ranged from 0.40 to 3.13 (K1 from 3.48 to 13.61 nM-1). Ro2 ranged from 2.88 to 17.25 (K2 from 0.03 to 0.65 nM-1). The most striking fact was the close positive correlation between the specific binding and the age of children (r = 0.914, P less than 0.001). These data suggest that the high growth velocity of young children, concomitant with the low plasma levels of IGF I which are physiological during infancy and early childhood, does not result from an increased binding of IGF I to cell receptors. PMID- 2974011 TI - Atrial natriuretic peptide in diabetic patients with microangiopathy. PMID- 2974012 TI - Molecular evidence for true isochromosome 21q. AB - In one family a duplicated 21q was shown to be a true isochromosome, which segregates from mosaic mother to non-mosaic child with full Down syndrome phenotype. Densitometric analysis of Southern blots, using probe pPW228C for the distal long arm of chromosome 21, indicated that the 21q duplication contains two copies of the allele detected by the probe. Maternal mosaic karyotype of 45,XX, 21/46,XX/46, XX,-21,+21i(21q) also suggested transverse mitotic centromere division as the origin of the 21q isochromosomes. Morphologic analysis of chromosome heteromorphisms strengthened this interpretation because the free 21 missing in the cell line with 45 chromosomes was also missing in cells with the isochromosome. In a second family the cytogenetic data also suggested transmission of an i(21q) from mosaic mother to non-mosaic Down syndrome child but molecular evidence did not prove identity of alleles in the duplicated chromosome 21. PMID- 2974013 TI - Increased methotrexate-induced chromosome breakage in patients with free trisomy 21 and their parents. AB - Increased susceptibility of chromosomes from peripheral blood lymphocytes to the antimetabolite methotrexate (2 X 10(-6) M) has been found in patients with free trisomy 21 and their parents (N = 14). The level of induced chromatid and chromosome breaks is lowest in normal controls intermediate in patients' mothers and fathers, and highest in trisomy 21 patients. The findings are viewed as a special type of cytogenetic polymorphism or as a defective chromosomal infrastructure, also in the parents of trisomic children. PMID- 2974014 TI - Comparative reactivity of anti-Ia monoclonal antibodies in man and laboratory animals. AB - Sixteen murine monoclonal antibodies (MAb), reactive with HLA-DR, DR + DP or DR + DQ, were tested, using indirect immunocytofluorescence, for their reactivity with peripheral blood mononuclear cells (PBMC) from the dog, cat, guinea pig, sheep, rabbit and rat. In addition, the MAb were evaluated for inhibitory activity in the canine mixed lymphocyte culture (MLC). Fourteen of 16 MAb reacted with canine PBMC. There was a greater tendency for DR + DP reactive MAb to inhibit canine MLC and subsequently react with PBMC of the guinea pig, sheep, and, cat. MAb failing to block the canine MLC were generally nonreactive with guinea pig PBMC (7 of 9 nonreactive) suggesting the guinea pig may be a useful model to study the functional relevance of specific Ia molecules. One MAb, H81.98.21 (reactive with HLA-DR) blocked canine MLC and reacted with PBMC from all species tested suggesting the determinant it recognized to be very well conserved in nature. PMID- 2974015 TI - Lipopolysaccharide-induced suppression of graft-versus-host reactivity in mice. AB - Reconstitution of lethally irradiated mice with spleen cells from donors that had been treated with lipopolysaccharide (LPS) intravenously and allogeneic spleen cells subcutaneously leads to a suppressed anti-host delayed-type hypersensitivity (DTH). Either donor injection alone proved to be ineffective. The state of suppression appeared to be antigen-specific, but, depending on the experimental conditions, also anti-host DTH to third-party alloantigens could be suppressed. The suppression was mediated by a population of Thy-1- suppressor cells that could also be induced in athymic nude mice. The suppressor cells specifically adhered to anti-kappa-coated plastic plates, but were not adsorbed by passage through a Sephadex G-10 column. Thus, it appears that the combined donor treatment with LPS and allogeneic spleen cells induces a population of B cells that can suppress anti-host immune reactivity. PMID- 2974016 TI - Autoreactive T cells in rheumatic disease. II. Function and specificity of an autoreactive T helper cell clone established from a HLA-B27+ reactive arthritis. AB - T cell lines were established by limiting dilution of peripheral blood (PBL) and synovial fluid lymphocytes (SFL) of a patient with HLA-B27+ reactive arthritis. Among these cell lines, the CD4 phenotype was dominant. Functionally, the majority of these cell lines exhibited helper activity for the immunoglobulin production by autologous B cells and proliferated in response to autologous mononuclear cells. In most cases, this autoreactive response was associated with alloreactivity. Only one cell line, the autoreactive CD4+ T cell clone, UA-S2, which was derived from the synovial fluid, proliferated in a highly specific manner in response to a determinant associated with MHC class II products present on autologous mononuclear cells. The restriction element was shown to be associated with DR molecules by inhibition experiments with monoclonal antibodies. Within the patient's family, the capacity of mononuclear cells to stimulate a proliferative response of UA-S2 segregated together with the HLA haplotype A2 or 32, B27, Cw1, DRw11 which was contributed by the patient's mother. UA-S2 proved to be a functional helper T cell clone. In the absence of additional antigen or mitogen, it induced IgG and IgM synthesis of autologous and family members' B cells. This helper activity of UA-S2 showed the same MHC restriction as the proliferative response. Although the patient's father also typed DRw11, this haplotype was not recognized by UA-S2. It is suggested that this autoreactive T cell clone detects a microheterogeneity of the serologically defined DRw11 haplotype. Indeed, typing of the patient's family members with cellular reagents established a difference between the two DRw11 haplotypes. PMID- 2974017 TI - Influence of age on conformation of rat muscle phosphofructokinase: analysis of pH dependent kinetics and limited proteolysis of enzyme. PMID- 2974018 TI - Calcium oxalate crystal growth studies in polyacrylamide gels: Part II--Influence of synthetic polypeptides and natural macromolecules on crystal aggregation. PMID- 2974020 TI - Long-term sickness absence due to back disorders in crane operators exposed to whole-body vibration. AB - In a retrospective (10-year) follow-up study, the incidence of at least one spell of sickness absenteeism of 28 d or longer in crane operators exposed to whole body vibration and a control group was investigated. In contrast to a previous study on permanent work disability in the same groups, no difference was observed in long-term sickness absenteeism because of lumbar disorders. Spells of sickness absence due to intervertebral disc disorders did last longer in the index group and also more disability pensions with this diagnosis were recorded in the index group. This indicates that these disorders particularly interfere with the work of a crane operator. Exposure to whole-body vibration and strained posture are considered to be responsible for this situation. PMID- 2974019 TI - Structure, function and clinical relevance of the low affinity receptor for IgE. PMID- 2974021 TI - In vitro effect of TP-1 (a calf thymic extract) on suppressor T-cell function of patients with autoimmune chronic active hepatitis. AB - Concanavalin A-activated T-lymphocyte suppression of IgG production was found to be significantly impaired in patients with untreated active autoimmune chronic hepatitis when compared to normals or patients with inactive disease. When the dose-response effect of TP-1, a calf thymic extract, on in vitro suppressor cell activity was assessed, lymphocytes from six out of eight patients with previously reduced suppressor cell function showed a significant improvement, while over a similar range the suppressor cell activity of most normal controls declined. These results support the possibility that defective immunoregulation in patients with autoimmune chronic active hepatitis may be related to a deficiency in thymic hormone levels. PMID- 2974022 TI - Different pathways for deoxyguanosine toxicity in T-lymphocytes of various developmental stages. AB - The basis of the selective cellular immunodeficiency which occurs in patients with purine nucleoside phosphorylase (PNP) deficiency still is not completely understood. We studied the mechanism of deoxyguanosine (dGuo) toxicity in proliferating lymphoid T-cells of different maturation stage, i.e. in T-cells of adult peripheral blood and cord blood and in CD3+ and CD3- subfractions of thymocytes. The mitogen-induced proliferation of T-cells from peripheral blood and cord blood and of CD3+ and CD3- subfractions of thymocytes. The mitogen induced proliferation of T-cells from peripheral blood and cord blood and of CD3+ thymocytes, as well as the spontaneous proliferation of CD3- thymocytes, are inhibited by dGuo. CD3+ and CD3- thymocytes are significantly more sensitive to dGuo than T-cells from peripheral blood or cord blood. Among the thymocyte subfractions CD3- thymocytes appeared to be extremely sensitive. In all cell types studied, inhibition of proliferation is accompanied by intracellular increases in both guanosine triphosphate (GTP) and deoxyguanosine triphosphate (dGTP) concentrations. By use of the PNP inhibitor 8-aminoguanosine, or the metabolites hypoxanthine or deoxycytidine, the metabolism of dGuo could be selectively directed to the formation of GTP or to dGTP. Based on the pattern of rescue from dGuo intoxication under these different metabolic conditions we conclude that in CD3- thymocytes dGuo toxicity is mediated by dGTP. In all other cell types studied GTP mediates dGuo intoxication. Altogether the results show that during the maturation from immature thymocytes to mature peripheral blood T cells a shift occurs in the pattern of dGuo toxicity since dGuo toxicity in the former is primarily caused via the dCyd kinase pathway, and in the latter mainly the degradation route is involved. Since in PNP deficiency mature T-cells do occur in the peripheral blood, we must conclude that some cells escape the stage of T-cell maturation in the thymus which is extremely sensitive to dGuo. Furthermore, the results imply that as far as T-cell development in the normal thymus is concerned, survival and death of cells might be regulated by local (deoxy) nucleoside availability. PMID- 2974023 TI - Quantitative enzyme histochemistry of rat foetal brain and trigeminal ganglion. AB - The increasing concern and the efforts in determining neurological effects in offsprings resulting from maternal exposure to xenobiotics are faced with several difficulties in monitoring damage to the central nervous system. In this paper, the efficiency of several enzyme histochemical reactions for analysing the forebrain and the trigeminal ganglia of rat foetuses are reported. Brains of 20 day-old Sprague-Dawley rat foetuses were frozen and analysed for 18 enzymes that had previously been used to monitor initial injury caused by toxic compounds in liver and other organs. Eight enzymes appeared suitable as histochemical markers for the functional integrity of different areas in brain and ganglia of rats exposed to xenobiotics. They were lactate, malate, glycerophosphate (NAD-linked), succinate, aldehyde and glucose 6-phosphate dehydrogenases, alpha glycerophosphate-menadione oxidoreductase and cytochrome c oxidase. The activities of the enzymes were determined by microphotometry and the arrangement of absorbances of the enzyme final reaction products into appropriate analytical tables is proposed as an efficient procedure for data analysis. PMID- 2974024 TI - Lack of keratan sulphate in the human notochord. AB - Intervertebral discs, formed from notochord cell expansions during embryogenesis, are known to contain proteoglycans bearing keratan sulphate chains. Keratan sulphate has previously been demonstrated in Xenopus and chick notochords and in human fetal cartilage. In contrast, we have been unable to demonstrate keratan sulphate in human fetal notochord using two monoclonal antibodies, MZ15 and 5-D 4. The contribution made by notochordal-derived keratan sulphate to the glycosaminoglycan content of the mature intervertebral disc may differ in man from that of other animal species. PMID- 2974025 TI - C-1027-AG, a selective antagonist of the macromolecular antitumor antibiotic C 1027. PMID- 2974026 TI - Effects of environmental design and police enforcement on violations of a handicapped parking ordinance. AB - This study presents two experiments that evaluate strategies to reduce violations of a handicapped parking ordinance. The first experiment compared effects of upright versus ground handicapped parking signs on percentage of intervals in which cars were parked illegally. Introducing upright signs produced an immediate reduction in the percentage of intervals of inappropriate use of parking spaces. The second experiment examined effects of a police enforcement program on percentage of intervals of inappropriate use of parking spaces and frequency of inappropriately parked cars. Results showed consistent reductions in percentage of intervals of inappropriate use and number of inappropriately parked cars compared with a control site where no enforcement program was introduced. Implications of the research data for law enforcement and public policy are discussed. PMID- 2974027 TI - Different expression of CD3 and CD22 in leukemic cells according to whether tested in suspension or fixed on slides. AB - The expression of antigens detected by monoclonal antibodies of the CD22 and CD3 groups was studied, by immunofluorescence (IF) on cell suspension and by immunoperoxidase (IP) on cells fixed on cytospin slides, in a range of B- and T cell malignancies. CD22 was demonstrated in the B-cell leukemias more frequently by IP than by IF; in B-lineage acute lymphoblastic leukemia (ALL) it was positive only by IP. In chronic lymphocytic leukemia and lymphoplasmacytic tumors approximately 40% of cases were CD22 positive by IP but few by IF. In other disorders of mature B cells, CD22 was positive by both methods in most cases. CD3 was demonstrated by IP in all 48 cases of T-cell leukemia and lymphoma regardless of the maturation stage (i.e., from early thymic to mature T cells). This was not the case by IF, where only 16% of T-ALL and 73% of cases with postthymic T-cell leukemias were recorded as positive. The greater reactivity of CD3 and CD22 by IP reflect the early cytoplasmic expression of the antigens in immature T and B cells, respectively, as well as the possible greater sensitivity of the IP method. These differences should be taken into account when describing results in lymphoid malignancies. CD3 was not expressed in cells from any of the B-cell leukemias, and likewise, CD22 was not demonstrated in any of the T-cell disorders, confirming that both reagents are highly specific for their respective lineages. PMID- 2974028 TI - AIDS in Arkansas. Update: June 1988. New Arkansas Department of Health regulation. PMID- 2974029 TI - Surgical management of post-traumatic prostatomembranous urethral strictures: a ten year experience. PMID- 2974030 TI - Significance of a positive antinuclear antibody: a clinical review. PMID- 2974031 TI - Nolie Mumey, a distinguished graduate. PMID- 2974033 TI - Transcriptional mapping and nucleotide sequence of the Escherichia coli fepA-fes enterobactin region. Identification of a unique iron-regulated bidirectional promoter. AB - The iron-controlled fepA and fes-entF transcripts from the Escherichia coli enterobactin gene complex are expressed divergently from a limited genetic region, thereby suggesting the existence of a single, possibly overlapping promoter junction for these genes. The nucleotide sequence of a 1,997-base pair HpaI fragment specific for this genetic region allowed for the identification of an 1,122-base pair open reading frame as the previously uncharacterized fes gene. Its product, Fes (approximately Mr 42,573) plays an essential but as yet ambiguous role in the release of ferric iron from the ligand. An additional small open reading frame of 216 nucleotides (encoding a potential product of calculated Mr 8,271) was also identified between fes and entF. A portion of the remaining nucleotide sequence defined a 320-base pair control region for both the fepA and fes-entF messages. Primer extension analyses placed the major in vivo transcription initiation sites to within 18 nucleotides of one another, thereby revealing a novel, extensively overlapping bidirectional promoter as well as long dual leader transcripts. This promoter region contains multiple overlapping nucleotide stretches which show strong homology to the consensus Fur repressor binding sequence, forms of which are found in all E. coli iron-regulated promoters characterized to date. PMID- 2974032 TI - Sedimentation equilibrium analysis of five lipocortin-related phospholipase A2 inhibitors from human placenta. Evidence against a mechanistically relevant association between enzyme and inhibitor. AB - Five proteins from human placenta capable of inhibiting pancreatic phospholipase A2 were purified. Two of these proteins were identified as lipocortins I and II. The other three proteins were immunologically distinct from lipocortins I and II and had apparent subunit molecular masses of 32, 33, and 73 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino acid sequence analysis of peptides produced by cyanogen bromide digestion indicated sequence homology of these proteins with lipocortin I and the heavy chain subunit of lipocortin II. Two of these proteins were identified as endonexin II and 67-kDa calelectrin. The third protein appears to be the human form of bovine endonexin I, also characterized as porcine protein II. Sedimentation equilibrium analysis of lipocortin I, endonexin I and II, and the 67-kDa calelectrin suggested monomer dimer equilibria with dissociation constants in the range of 0.33-1.3 X 10(-3) M and monomer molecular masses of 38,050, 36,400, 36,850, and 73,610 Da, respectively. Self-association of lipocortin II was described by dimerization of a protomer (K12 = 5.3 x 10(-7) M), followed by an indefinite self-association of the dimer (isodesmic dissociation constant, Kiso = 3.6 x 10(-6) M). The protomer molecular mass was 48,800 Da, consistent with a heterodimeric structure composed of one heavy (38,600 Da) and one light (10,944 Da) chain as previously characterized for lipocortin II. Sedimentation equilibrium analysis of mixtures of individual protein inhibitors and purified pancreatic phospholipase A2 indicated weak association between enzyme and inhibitor (Kd greater than or equal to 3 x 10(-5) M), insufficient to account for the observed inhibition of enzyme activity. PMID- 2974034 TI - Proteolytic processing of mullerian inhibiting substance produces a transforming growth factor-beta-like fragment. AB - Mullerian inhibiting substance (MIS) is a differentiation factor that causes the Mullerian duct to regress during the development of the male reproductive tract. The active form is a disulfide-linked dimer consisting of two identical 70-kDa subunits. Recently, the amino acid sequence for MIS was deduced from its gene sequence and revealed that the carboxyl-terminal region shares homology with transforming growth factor (TGF)-beta. Since TGF-beta is produced as a large latent precursor that requires proteolytic activation for activity, we sought to determine if MIS might undergo a similar processing event. Here we demonstrate that typically 5 to 20% of the protein in MIS preparations is cleaved at a site 109 amino acids from the carboxyl terminus. Concurrent cleavages from both chains of the MIS dimer produces a 25-kDa TGF-beta-like fragment and a high molecular mass complex derived from the amino terminus of the protein. Although the two fragments are noncovalently linked, they remain tightly associated after cleavage, and thus are structurally organized like TGF-beta within its precursor. The same cleavage products also can be generated by limited proteolysis with plasmin, which provides a simple method for converting the entire preparation into the cleaved form. The plasmin-digested MIS is fully active in the organ culture assay. PMID- 2974035 TI - Activation of a cytosolic serine protein kinase by epidermal growth factor. AB - Exposure of A-431 cells to epidermal growth factor (EGF) results in a rapid enhancement (approximately 10-fold) of cytosolic serine protein kinase activity. The increase in serine kinase activity may be detected using a number of peptide and protein substrates. Enhancement of kinase activity occurs within 1 min of exposure of the cells to EGF and reaches a maximum in 5 min. Similar results were obtained with a variety of cell lines. We have partially purified the EGF activated kinase from A-431 cells. It has an apparent molecular mass of approximately 100 kDa by gel filtration. One distinguishing property of the enzyme is its sensitivity to inhibition by micromolar quantities of polyarginine; polylysine has no effect. The EGF-activated kinase is unaffected by cyclic nucleotides, Ca2+/calmodulin, Ca2+/diolein/phosphatidylserine, or heparin. The enhancement of cytosolic serine kinase activity in A-431 cells appears to be an early event in cell "activation" by a number of biological response modifiers including EGF, bradykinin, 12-O-tetradecanoylphorbol-13-acetate, and histamine. PMID- 2974036 TI - Neocarzinostatin-induced DNA base release accompanied by staggered oxidative cleavage of the complementary strand. AB - Treatment of an end-labeled DNA restriction fragment with the nonprotein chromophore of neocarzinostatin induced lesions which, after treatment with endonuclease IV or putrescine, were expressed as site-specific double-strand breaks. Analysis of the termini at cleavage sites in each strand showed that the neocarzinostatin-induced lesions consisted of an apurinic/apyrimidinic site plus a closely opposed break in the complementary strand. The break always occurred opposite the base two positions upstream from the apurinic/apyrimidinic site and had the 3'-phosphate and 5'-aldehyde termini characteristic of neocarzinostatin induced breaks. This positioning suggests that neocarzinostatin simultaneously attacks two DNA sugars on opposite edges of the minor groove. The sequence specificity for formation of apurinic/apyrimidinic sites with closely opposed breaks reflected that of neocarzinostatin-induced mutagenesis. The potent mutagenicity of these lesions may be attributable to the presence of closely opposed damage in both DNA strands. PMID- 2974037 TI - 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid inhibits CD3-T cell antigen receptor-stimulated Ca2+ influx in human T lymphocytes. AB - Stimulation of the CD3-T cell antigen receptor complex on T lymphocytes results in a rapid rise in intracellular calcium from both intra- and extracellular sources. The former is thought to be released from the endoplasmic reticulum in response to inositol trisphosphate, while the latter enters the cells through a membrane potential-sensitive transporter (Oettgen, H. C., Terhorst, C., Cantley, L. C., and Rosoff, P. M. (1985) Cell 40, 583-590). In this report we show that the stilbene disulfonate, DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid), inhibited the ability of monoclonal anti-CD3 complex antibodies to stimulate an influx of calcium in the human T lymphocyte cell line, Jurkat. DIDS had no effect on either antibody binding to the receptor or receptor-stimulated phosphatidylinositol turnover. The Ki was approximately 25 microM in the presence of extracellular Cl- and 10 microM when labeling was performed in the absence of Cl-, suggesting that DIDS was competing with Cl- for binding to the cell membrane. The reduced form of DIDS, dihydroDIDS, was only 50% as effective as DIDS itself, and the monoisothiocyanate stilbene, 4-acetamido-4' isothiocyantostilbene-2,2'-disulfonic acid, was totally ineffective, even to concentrations of 0.750 mM. Removal of extracellular Cl- also inhibited the antibody-stimulated influx of calcium. These data suggest that the function of the CD3-T cell receptor-activated calcium channel/transporter may be dependent on or regulated by extracellular Cl-. PMID- 2974038 TI - Fibronectin's amino-terminal matrix assembly site is located within the 29-kDa amino-terminal domain containing five type I repeats. AB - Fibronectin is organized into disulfide cross-linked, insoluble pericellular matrix fibrils by fibroblasts in vitro. Two sites, the Arg-Gly-Asp-Ser-containing cell attachment domain and a site located in the first 70 kDa of fibronectin, are required for matrix assembly. The first 70 kDa of fibronectin contain two structural motifs termed type I and type II homologies, which are repeated nine and two times, respectively. Previous work has implicated the amino-terminal region and the carboxyl terminus containing three type I repeats in matrix assembly, suggesting that type I repeats possess binding activity essential for fibronectin matrix assembly. To test this hypothesis, we developed a sensitive capture immunoassay to quantify insoluble matrix fibronectin and tested a panel of fibronectin fragments, containing all of the type I repeats found in the intact protein, for their ability to inhibit matrix assembly. Only fragments containing the first five type I repeats inhibited fibronectin matrix assembly, although sequences carboxyl-terminal to this domain enhanced this activity. Additional evidence for the specific recognition of the amino-terminal type I repeats by matrix assembling cells was found when the reversible, detergent sensitive binding of a 125I-labeled fragment containing the first five type I repeats (29 kDa) to cell monolayers was studied. Only monolayers of cell lines that incorporate fibronectin into a fibrillar matrix specifically bound 125I labeled 29 kDa. Binding of the radiolabeled amino-terminal fragment to matrix forming cells was inhibited by unlabeled fragments containing the first five type I repeats but not by unlabeled fragments containing the remaining seven type I repeats. Matrix assembly is therefore not a generalized property of type I repeats. Rather, a critical site is located within the first 29 kDa of fibronectin. PMID- 2974040 TI - Mutations at twelve independent loci result in absence of outer dynein arms in Chylamydomonas reinhardtii. AB - 35 strains of Chlamydomonas mutant missing the entire outer dynein arm were isolated by screening slow-swimming phenotypes. They comprised 10 independent genetic loci (odal-10) including those of previously isolated mutants oda38 and pf28. The 10 loci were distinct from pf13 and pf22, loci for nonmotile mutants missing the outer arm. These results indicate that at least 12 genes are responsible for the assembly of the outer dynein arms. There were no mutants lacking partial structures of the outer arm, suggesting that lack of a single component results in failure of assembly of entire outer arms. Temporary dikaryons derived from mating of two different oda strains often, but not always, recovered the wild-type motility within 2 h of mating. Hence, outer arms can be transported and attached to the outer doublets independently of flagellar growth. PMID- 2974041 TI - Response of aggregating chick corneal cells to modifiers of N-linked oligosaccharides, endoglycosidase H and deoxymannojirimycin. AB - Chick corneal epithelium takes on its mature conformation between 11 and 16 days of incubation. Earlier work has shown that desmosome frequency increases during this period, reaching its highest rate at 15 1/2 days. In the present report aggregation rates of cells from embryos of 11 days and those of 15 1/2 days are compared. Younger cells, which form fewer desmosomes, aggregate at a more moderate rate than older cells. In addition, younger cells bind less concanavalin A (ConA) than older cells. To determine if increase in ConA binding could be related to these cellular responses, aggregating cells were exposed to endoglycosidase H (EndoH) and to deoxymannojirimycin. This treatment should permit comparison of the response of cells that have a normal complement of N linked oligosaccharides with those that have reduced high-mannose or complex type sugars. The effectiveness of EndoH under the conditions used was confirmed by failure of treated glycoprotein after separation by SDS-PAGE and electroblotting to bind ConA. Aggregation rates of both older and younger cells were unaffected, as measured by disapperance of single cells, though older cells formed somewhat smaller aggregates at the highest dosage used. Desmosome formation was markedly reduced in the presence of the enzyme, even in the absence of other changes in the fine structure. At the highest dose of the enzyme the fine structure of older but not younger cells showed indications of blockage of transport. Deoxymannojirimycin appears to cause a build-up of high-mannose groups, since treated cells showed increased incorporation of [3H]mannose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974039 TI - Selective degradation of T cell antigen receptor chains retained in a pre-Golgi compartment. AB - We have examined the fate of newly synthesized T cell antigen receptor (TCR) subunits in a T cell hybridoma deficient in expression of the clonotypic beta chain. Synthesis and assembly of the remaining chains proceed normally but surface expression of TCR chains is undetectable in these cells. A variety of biochemical and morphological techniques has been used to show that the TCR chains in these cells fail to be transported to any of the Golgi cisternae. Instead, they are retained in a pre-Golgi compartment which is either part of or closely related to the endoplasmic reticulum. The CD3-delta chain is degraded by a non-lysosomal process that is inhibited at temperatures at or below 27 degrees C. By contrast, the remaining chains (CD3-epsilon, CD3-gamma, and zeta) are very stable over 7 h. We propose possible mechanisms that may explain the differential fate of TCR chains retained in a pre-Golgi compartment. PMID- 2974042 TI - Intercellular interactions in myelin-specific autoimmunity. PMID- 2974043 TI - T-cell subsets in the cerebrospinal fluid in the course of inflammatory neurological diseases. PMID- 2974044 TI - Structural basis of antigen recognition by T lymphocytes. Implications for vaccines. PMID- 2974045 TI - Immunological abnormalities in human immunodeficiency virus (HIV)-infected asymptomatic homosexual men. HIV affects the immune system before CD4+ T helper cell depletion occurs. AB - To investigate the effect of persistent HIV infection on the immune system, we studied leukocyte functions in 14 asymptomatic homosexual men (CDC group II/III) who were at least two years seropositive, but who still had normal numbers of circulating CD4+ T cells. Compared with age-matched heterosexual men and HIV negative homosexual men, the CD4+ and CD8+ T cells from seropositive men showed decreased proliferation to anti-CD3 monoclonal antibody and decreased CD4+ T helper activity on PWM-driven differentiation of normal donor B cells. Monocytes of HIV-infected homosexual men showed decreased accessory function on normal T cell proliferation induced by CD3 monoclonal antibody. The most striking defect in leukocyte functional activities was observed in the B cells of HIV-infected men. B cells of 13 out of 14 seropositive men failed to produce Ig in response to PWM in the presence of adequate allogeneic T-helper activity. These findings suggest that HIV induces severe immunological abnormalities in T cells, B cells, and antigen-presenting cells early in infection before CD4+ T cell numbers start to decline. Impaired immunological function in subclinically HIV-infected patients may have clinical implications for vaccination strategies, in particular the use of live vaccines in groups with a high prevalence of HIV seropositivity. PMID- 2974047 TI - Characterization of the intrinsic fibrinolytic properties of pro-urokinase through a study of plasmin-resistant mutant forms produced by site-specific mutagenesis of lysine(158). AB - Two plasmin-resistant mutant forms of pro-urokinase (pro-UK) constructed by site directed mutagenesis of Lys158 to Val158 and Met158 were used to evaluate the intrinsic enzymatic and fibrinolytic properties of pro-UK as distinct from those of its two-chain UK (TC-UK) derivative. Both mutants, while resistant to plasmin activation, were as sensitive as pro-UK to degradation by thrombin. Since thrombin cleaves a peptide bond only two residues from the activation site, the integrity of this loop was maintained in the two mutants. The amidolytic and plasminogen-activating activities of the mutants averaged 0.14 and 0.12% that of TC-UK, respectively. The fibrin plate activities were 2,400 IU/ml and 700 IU/mg for the Met158 and Val158 mutants or about 1.5% that of TC-UK. These findings attest to a discrete but low intrinsic activity for pro-UK and suggest that the higher values reported in the literature may be related to UK contaminants or plasmin-induced TC-UK generation during the assay. Clot lysis by the mutants required doses greater than 100-fold higher than those of pro-UK to induce a comparable effect. From this it appears that pro-UK activation is a major determinant of the rate of clot lysis occurring with pro-UK. Clot lysis by the mutants was potentiated by plasmin pretreatment of the fibrin and by the addition of small amounts of TC-UK or tissue plasminogen activator (t-PA). Combinations of t-PA and the mutants were synergistic in their fibrinolytic effects. These findings mirror those previously obtained with pro-UK. We concluded that the previously described potentiation of pro-UK-induced clot lysis by UK or t-PA is mediated primarily by pro-UK itself rather than by a promotion of its activation. PMID- 2974048 TI - Inhibition of the anticoagulant activity of protein S by prothrombin. AB - Protein S is a vitamin K-dependent protein cofactor to the anticoagulant, activated protein C (APC). This study examines the inhibition of human protein S anticoagulant activity by prothrombin. In the absence of protein S, the anticoagulant activity of APC measured in a Factor Xa recalcification time, was comparable using normal or plasma adsorbed with Al(OH)3. Protein S was an effective cofactor to APC in Al(OH)3-adsorbed plasma, but was significantly less active in normal plasma. Analysis of the difference in the two plasmas revealed that normal plasma contained an inhibitor to the anticoagulant activity of protein S that was removed by Al(OH)3 adsorption. Purification of this inhibitory activity demonstrated that it was mediated by the vitamin K-dependent protein, prothrombin. Prothrombin purified by conventional techniques caused immediate, dose-dependent inhibition of the cofactor activity of protein S in the presence of phospholipids or platelets, but had no effect on the anticoagulant activity of APC. The inhibition was demonstrable using a Factor Xa recalcification time, and studies of the rates of inactivation of purified Factor Va. Increasing concentrations of protein S overcame the inhibition by prothrombin and kinetic analysis of the interaction demonstrated that prothrombin acted as a competitive inhibitor to protein S. Immunoabsorption of prothrombin from plasma using immobilized antiprothrombin antibodies was associated with the complete removal of the protein S inhibitory activity. We conclude that the anticoagulant activity of protein S is modulated by prothrombin and that this may represent another regulatory mechanism of the natural anticoagulant system. PMID- 2974049 TI - Treatment of rapidly destructive periodontitis in Papillon-Lefevre syndrome. Laboratory and clinical observations. AB - This paper reports the successful treatment of the periodontal component of the Papillon-Lefevre syndrome in 2 siblings. Treatment consisted of extractions of periodontally-involved teeth under antibiotic therapy. The exogenous source of the suspected pathogenic A. actinomycetemcomitans in this family, identified as a pet dog, was also treated with antibiotics. Assessments of monocyte function and levels of serum antibodies against A. actinomycetemcomitans were performed prior to an immediately after treatment. Microbiological screening of subgingival dental plaque of the PLS patients as well as the rest of the household members, including the dog, was performed every month during the study period. Remission of the rapidly destructive periodontitis, as well as an earlier-described monocyte dysfunction in these two PLS patients occurred concomitantly with the eradication of A. actinomycetemcomitans from the family. 15 months after this treatment, the children still had no signs of periodontitis or of A. actinomycetemcomitans infection. The legitimacy with which PLS is defined as a disease entity is discussed and questioned. PMID- 2974050 TI - "Immunoregulatory effects of cimetidine: inhibition of suppressor cell effector function in vivo". AB - The effects of cimetidine upon suppressor cell effector function in a well studied murine model of contact hypersensitivity were examined. Intravenous inoculation of BALB/c mice with DNP-coupled syngeneic spleen cells induced the production of DNP-specific suppressor cells which was demonstrated by a reduction in ear swelling after contact sensitization with 1-fluoro-2, 4-dinitrobenzene (DNFB) following transfer of spleen and lymph node cells to naive syngeneic recipients. Cimetidine treatment of mice who had also received suppressor cells eliminated the manifestation of suppressor cell activity as measured by the development of normal immunologic response following contact sensitization with DNFB. While all the groups receiving cimetidine showed restoration of delayed hypersensitivity, the maximum effect was seen when 50 mg/kg of cimetidine was administered on Day 5 (day of challenge). These results indicate that, in addition to its previously described inhibitory role in suppressor cell induction, cimetidine is also capable of inhibiting suppressor cell effector function. The involvement of histamine in both these processes in vivo is also suggested. PMID- 2974046 TI - Binding of Lys-plasminogen to monocytes/macrophages. AB - The ability of mononuclear phagocytes to assemble and activate components of the fibrinolytic system on their surfaces may be crucial in effecting an efficient inflammatory response. Lys-plasminogen, the plasmin modified form of this zymogen, was found to bind specifically and with high affinity to murine peritoneal macrophages and to cells of the human monocytoid line U937. This modified plasminogen has been shown to be a more efficient substrate for plasminogen activators than native Glu-plasminogen. Binding was lysine binding site dependent, rapid and reversible. In contrast, although native Glu plasminogen bound specifically to these cells, affinity was low. Lys-plasminogen inhibited the binding of Glu-plasminogen but the opposite was not true. Molecular analysis of the bound ligands indicated that Glu-plasminogen was converted to Lys plasminogen and Lys-plasminogen to plasmin on the cell surface but not in the supernatant. Peritoneal macrophages from patients with indwelling catheters and tissue macrophages in chronic inflammatory lesions were shown to express immunologically identified Lys-plasminogen on their surfaces. Therefore binding and surface activation of kinetically favored Lys-plasminogen may provide an important physiological mechanism for localizing proteolytic activity on the surface of inflammatory cells. PMID- 2974051 TI - The role of soluble acids from oxidative-hydrolytic breakdown of coal in the development of the fibrous process in anthracosis. AB - The oxidative-hydrolytic mechanism of interaction of coal with tissue medium and the formation of humic acids (HA) and polycarboxylic acids (PCA) from coal in the organism were demonstrated. The composition of PCA obtained from the coal withdrawn from the organism of animals was investigated by gas-liquid chromatography by means of which 28 acids of the aromatic series were identified. The method of elemental spectral analysis revealed the transition of a number of macro- and microelements into the medium of the organism. It has been shown experimentally that the decrease in the mass of coal correlates with the fibrogenic process. The conclusion has been drawn that the soluble products of the oxidative-hydrolytic breakdown of coal in the organism are among the fibrogenic factors of pneumosclerosis in anthracosis. The initial mechanism in the development of fibrosis is the damage caused by polycarboxylic acids to the microcirculatory bed of the lungs and the structures of the aerohematic barrier. PMID- 2974052 TI - The effect of atrial natriuretic factor on urinary albumin and beta 2 microglobulin excretion in man. AB - The effect of a 20-min infusion of atrial natriuretic factor (ANF) 99-126, 0.4 microgram/kg per min, on both urinary albumin and beta 2-microglobulin excretion was examined in nine normal male subjects during stable water diuresis. ANF caused a rise in urinary albumin in excretion (from 4.19 +/- 0.66 to 13.49 +/- 3.07 ng/min, P less than 0.01) without any significant change in either creatinine clearance of beta 2-microglobulin excretion. These findings suggest that ANF may enhance glomerular permeability to albumin in man. PMID- 2974054 TI - Helper T lymphocytes which recognize the MHC class I alloantigens in vivo are CD4+CD8-1. AB - To determine the CD4 or CD8 phenotype of the Th lymphocyte which recognizes in vivo the MHC class I alloantigens, B10 recombinant mice were treated with anti CD8 or anti-CD4 mAb and immunized with lymphoid cells from donors differing in the K or D region of the MHC. Alloantibodies were evaluated by a 51Cr-release assay or by indirect immunofluorescence. The production of IgG anti-Dd and anti Kk alloantibodies was increased by the deletion of the CD8+ and absent in mice depleted of the CD4+ subset. These experiments indicate that the helper influence elicited by the recognition of a MHC class I alloantigen in vivo is due to cells of the CD4+CD8- phenotype. PMID- 2974055 TI - The relationship between infiltrating CD4+ lymphocytes, activated eosinophils, and the magnitude of the allergen-induced late phase cutaneous reaction in man. AB - The phenotype and activation status of leukocytes infiltrating human late phase allergic skin reactions were investigated by immunocytochemistry using a panel of mAb. Late phase skin reactions were induced in atopic individuals by intradermal challenge with allergen extract (grass pollen or house dust mite) and skin biopsies were obtained 6, 24, or 48 h after challenge. Cryostat sections were examined for evidence of infiltration and activation of T cells and eosinophils. Biopsies from saline-challenged control sites were used for comparison. A substantial number of CD3+ cells were observed close to the dermal capillaries. The number of CD4+ cells showed a similar pattern but few CD8+ cells were seen. The ratio of CD4 and CD8 subsets in tissue did not relate to the CD4/CD8 ratio in the peripheral blood. Two observations suggested that T cells had become activated: first, a small number of infiltrating cells bore receptors for IL-2, and second, there was indirect evidence of IFN-gamma secretion, demonstrated by increased expression of HLA-DR on endothelial cells and de novo expression of CD4 Ag on epidermal Langerhans cells. Activated eosinophils were detected in most of the allergen-challenged biopsies. The late phase reaction diameter at 6 h correlated with the number of activated eosinophils at 48 h (r = 0.61, p = 0.05), but not with infiltration by T cells. There was a strong correlation between the numbers of CD4+ cells and activated eosinophils at 24 h (r = 0.94, p less than 0.001). These findings suggest that interactions between T lymphocytes and eosinophils in reactions to airborne allergens may be more important than has previously been recognized. PMID- 2974053 TI - Regulation of antigen-presentation-I. IFN-gamma induces antigen-presenting properties on B cells. AB - B cells require activation to efficiently present Ag to T cells. In agreement with this earlier observation we show that live, mitomycin C-treated B cells, but not B cells fixed in paraformaldehyde, stimulated the growth of allogeneic T cells in the primary MLR. However, if B cells were cultured with anti-Ig antibodies and IFN-gamma before fixation they acquired excellent T cell stimulatory activity. Neither reagent alone conferred this novel co-stimulatory function on the B cell surface. The activity induced by both stimuli was not attributed to an increase expression of class II-MHC molecules or IL-1. IL-2 or IL-4, in combination with anti-Ig, also induced B cell stimulatory activity, but were less effective than IFN-gamma. TNF failed to stimulate B cells, but synergized with IFN-gamma in the induction of this activity. These studies therefore demonstrate an important role for lymphokines in modulating B cell Ag presenting activity as well as the acquisition by B cells of a novel co stimulatory surface activity. PMID- 2974056 TI - IL-4 (B cell stimulatory factor 1) overcomes Fc gamma receptor-mediated inhibition of mouse B lymphocyte proliferation without affecting inhibition of c myc mRNA induction. AB - Mouse B cells are stimulated to proliferate by Fab'2 fragments of rabbit anti mouse Ig antibodies. Proliferation is inhibited, however, in the presence of IgG anti-mouse Ig. We have previously shown that this inhibition is mediated by binding of the IgG anti-Ig to receptors for Fc gamma R on B cells. This report describes conditions under which IgG anti-mu or anti-delta will induce proliferation despite Fc gamma R engagement. Culture supernatants of Con A stimulated, Il-4-secreting Th cell lines, but not of Il-2-secreting Th cell lines, will co-stimulate with IgG anti-Ig to induce small B cells to incorporate [3H]TdR. This co-mitogenic activity is inhibitable by anti-IL-4 antibodies and can also be induced by Il-4 affinity purified from the T cell supernatants or by supernatants containing rIl-4. B cells precultured with Il-4 for 18 h, while still expressing normal levels of Fc gamma R, also proliferate to IgG anti-Ig. We have previously shown that Fc gamma R-mIg cross-linking will inhibit mIg dependent increases in c-myc mRNA levels. We investigated whether Il-4 allows B cells to respond to IgG anti-Ig by elevating c-myc. The data show that Il-4 has little effect on c-myc mRNA levels in either IgG or Fab'2 anti-Ig-containing cultures. PMID- 2974058 TI - A monoclonal antibody that inhibits secretion from rat basophilic leukemia cells and binds to a novel membrane component. AB - Rat basophilic leukemia (RBL-2H3) cells, like mast cells and basophils, carry monovalent membrane receptors with high affinity for IgE (Fc epsilon R). Cross linking of these receptors provides the immunologic stimulus which initiates a series of biochemical events, culminating in secretion of inflammatory mediators. In an attempt to identify membrane components involved in the stimulus-secretion coupling of these cells, hybridomas were produced from splenocytes of mice immunized with intact RBL-2H3 cells. Here we report the production of a mAb (designated G63) that inhibits the Fc epsilon R-mediated secretion from RBL cells. At low degrees of Fc epsilon R aggregation, the mAb G63-induced inhibition may be complete, whereas at the maximum of secretion the inhibition is in the range of 30 to 40%. The relative degree of inhibition of secretion is dependent on the dose of mAb G63. Furthermore, inhibition requires the bivalency of G63, as the Fab fragments are inactive. The number of antigenic epitopes recognized by G63 per RBL-2H3 cell is 1.8 x 10(4) epitopes/cell, as determined by direct binding studies of 125I-labeled Fab fragments of G63. This number is 20 to 30 times smaller than that of Fc epsilon R on the same cells. The membrane component to which G63 binds has been identified by immunoprecipitation as a glycoprotein with an apparent Mr of 58 to 70 kDa. All of these results, and the fact that no competition for binding to RBL cells between mAb G63 and IgE can be resolved, indicate that mAb G63 binds to a membrane component which is distinct from the Fc epsilon R. mAb G63 suppresses the Fc epsilon R-mediated rise in cytoplasmic concentration of free Ca2+ ions, known to be one of the biochemical signals involved in the stimulus-secretion coupling in RBL-2H3 cells. G63 does not affect, however, the degranulation induced by the Ca2+ ionophore A23187. Therefore, mAb G63 probably exerts its inhibitory effect on a step preceding the rise in cytoplasmic free Ca2+. Thus, mAb G63 defines a previously unidentified membrane component that is involved in one of the early steps of the RBL-2H3 activation mediated by their Fc epsilon R. PMID- 2974059 TI - [Long-term observation of the immunogenicity of hepatitis B vaccine in hospital personnel]. PMID- 2974057 TI - Functional and biochemical characterization of rat bone marrow derived mast cells. AB - The functional and biochemical characterization of rat bone marrow derived mast cells (RBMMC) confirms both species-related differences between rat and mouse bone marrow-derived mast cells (MBMMC) as well as mast cell heterogeneity in a single species. Such RBMMC have the staining characteristics of mucosal mast cells and contain the mucosal mast cell protease. The RBMMC release the preformed granule mediator beta-hexosaminidase both in response to immunologic stimulation with 200 ng Ag (net release 15.8 +/- 3.8%) and in response to 1 microM calcium ionophore A23187 (net release 21.8 +/- 6.8%). However, compound 48/80, substance P, and somatostatin did not induce mast cell degranulation. In experiments with optimal beta-hexosaminidase release, the RBMMC generated similar quantities of the newly formed arachidonic acid metabolites leukotriene C4 and PGD2 when stimulated with either Ag or calcium ionophore A23187. The RBMMC incorporate [35S]sulfate into proteoglycans consisting of 90% chondroitin sulfates and 10% heparin. The chondroitin sulfates were comprised of chondroitin 4 sulfate and chondroitin sulfate diB sulfated disaccharides in a ratio of 4/1. Although we show that RBMMC and MBMMC share a low histamine content, functional IgE receptors and unresponsiveness to cromolyn and selective secretagogues (compound 48/80, substance P, and somatostatin), we also provide evidence that RBMMC differ from MBMMC in their profile of newly generated mediators, preformed granule proteoglycan, and lack of proliferative response to mouse IL-3. PMID- 2974061 TI - [Comparative study of transcutaneous oxygen pressure and laser- Doppler flow in the feet of 15 healthy subjects]. AB - Measurement of TcPO2 in arterial diseased patient is a reliable and reproductible method for evaluation in stages III and IV of the degree of tissue ischemia which can serve to establish the viable prognosis of the limb. One of the disadvantages of this technique is the relatively long time taken. By contrast the Doppler laser, a more recently introduced technique, can be used to measure superficial cutaneous flow. One of its advantages is the rapidity of its instantaneous measurements. Before testing this technique in arterial disease sufferers it was felt to be of interest to determine the possible existence in the healthy subject of a correlation between TcPO2 and laser-Doppler flow. The study involved 15 healty subjects, 8 men and 7 women, with a mean age of 24.2 years. TcPO2 and laser-Doppler measurements were carried out in the fore-foot at 38 degrees and 44 degrees, under basal conditions and then after ischemia each time for 5 minutes, finally followed by a 10-minute 100% hyperoxia test. There was only a slight correlation at 38 degrees between the two methods and no correlation was found at 44 degrees. With oxygen therapy TcPO2 increased considerably and there was a nonsignificant decrease in laser-Doppler flow. In total, the two methods did not truly explore the same microcirculatory data and clinical studies would appear necessary to determine whether laser-Doppler flow measurements in the arterial disease patient may prove to be as useful as those of TcPO2. PMID- 2974060 TI - Fenretinide does not block visual pigment formation in the rat. AB - Abnormal retinal function has been reported in patients receiving the synthetic retinoid fenretinide. In this study, the vitamin A deprived rat was used as a model to test the possibility that fenretinide is interfering with the vitamin A transport system and/or blocking the normal uptake of 11-cis retinal into the visual pigment. Assessing retinal function by electroretinogram measurements and visual pigment formation by competition studies, our data demonstrate normal uptake of vitamin A and normal visual pigment formation in the rat after treatment with the drug. PMID- 2974062 TI - [Transluminal percutaneous angioplasty by thermal lasers. 20 cases of peripheral artery occlusion]. AB - The authors report their experience of laser thermal angioplasty in a series of 20 patients. An argon laser with a 1.5 or 2.0 millimeter diameter metalcapped optical fiber (Hot Tip Trimedyne Inc.) was used; the laser probe was pushed to the occlusion point; after a channel was produced through the obliteration, a conventional percutaneous balloon angioplasty was performed. The indications were: 4 primitive iliac occlusions, 15 femoral/popliteal occlusions and 1 subclavian obliteration. The immediate results were as follows (table I): 75% overall recanalization (50% at the iliac level, 87% at the femoral/popliteal level and failure at the subclavian level); 5 notable complications were observed (2 limb ischaemia, 2 voluminous inguinal haematoma and 1 aortic cross perforation) without clinical sequelae. The short term follow-up (1 to 8 months) of the successfully treated patients (table II) shows 100% permeability at the iliac level and 77% at the femoral/popliteal level. We conclude that laser thermal angioplasty is a very attractive method in the treatment of primitive iliac and femoral/popliteal occlusions. The well-defined indications must form the subject of a closely discussion between the angiologist, the vascular surgeon and the interventional radiologist. PMID- 2974063 TI - The lysine residue in the membrane-spanning domain of the beta chain is necessary for cell surface expression of the T cell antigen receptor. AB - The TCR is a complex receptor composed of seven polypeptide chains consisting of a ligand-binding subunit, Ti, and a putative signal-transducing subunit, CD3. Phylogenetically conserved charged amino acid residues within the membrane spanning domains present in all seven chains of the TCR have been proposed to be important in the association between Ti and CD3. Using a Ti beta chain-deficient mutant of the cell line Jurkat, site-directed mutagenesis and transfection of Ti beta chain cDNA was performed to assess the importance of the lysine residue at position 290 within the membrane-spanning domain of the Ti beta chain to expression of the TCR complex. These studies demonstrated that the lysine residue, and not simply conservation of either basic charge or secondary structure, is important at this position. PMID- 2974064 TI - Functional heterogeneity of L3T4+ T cells in MRL-lpr/lpr mice. L3T4+ T cells suppress major histocompatibility complex-self-restricted L3T4+ T helper cell function in association with autoimmunity. AB - The present study demonstrates in MRl-lpr/lpr autoimmune mice an age-dependent loss of MHC-self-restricted function by L3T4+ Th. This defect is not present in age-matched, congenic MRL-+/+ spleen cells and appears to be due to the presence of suppressor cells that are selective for L3T4+ Th and not for Lyt-2+ Th. Surprisingly, the suppressor cells are also L3T4+ T cells and can suppress the IL 2 production of congenic MRL/+ L3T4+ Th to MHC-self-restricted antigens. These data support the idea of functional specialization within the L3T4+ population of T cells. Because L3T4+ suppressor cells are detected late in the course of autoimmunity, we interpret their presence not as a primary initiating event in the development of autoimmunity, but rather as a compensatory mechanism. Additionally, similar suppression of L3T4+ Th function has also been reported in a murine graft-vs.-host model of autoimmunity, suggesting that the suppressor cells represent an immunoregulatory mechanism that is a common feature of autoimmunity. Since excessive class II-restricted Th activity for B cells has been reported for both models of autoimmunity, L3T4+ suppressor cells may represent an attempt to down regulate such excessive Th activity. These findings may be relevant to human autoimmune diseases, such as systemic lupus erythematosus, in which B cell hyperactivity is also associated with reduced IL-2 production by Th. PMID- 2974065 TI - Immunologically mediated regression of a murine lymphoma after treatment with anti-L3T4 antibody. A consequence of removing L3T4+ suppressor T cells from a host generating predominantly Lyt-2+ T cell-mediated immunity. AB - This study shows that intravenous injection of 1 mg of anti-L3T4 mAb (GK1.5) into thymectomized mice bearing the syngeneic L5178Y lymphoma results, after a delay of 2-3 d, in complete regression of this tumor and in long-term host survival. A flow cytofluorometric examination of the spleen cells of mAb-treated mice revealed that antibody treatment resulted in the elimination of greater than 98% of L3T4+ T cells, but had no effect on the Lyt-2+ T cells subset. Tumor regression was immunologically mediated, because L5178Y lymphoma cells were shown to be L3T4-, and regression of the tumor failed to occur in mice that had been lethally irradiated before anti-L3T4 mAb was given. Tumor regression was mediated by tumor-sensitized Lyt2+ T cells, as evidenced by the finding that treatment of tumor-bearing mice with anti-Lyt-2 mAb alone, or in combination with anti-L3T4 mAb, resulted in enhancement of tumor growth and a significant decrease in host survival time. Moreover, the spleens of mice whose tumors were undergoing regression in response to anti-L3T4 mAb treatment contained Lyt-2+ T cells capable, on passive transfer, of causing regression of a tumor in recipient mice. These results can be interpreted as showing that removal of tumor-induced L3T4+ suppressor T cells results in the release of Lyt-2+ effector T cells from suppression, and consequently in the generation of enough Lyt-2+ T cell-mediated immunity to cause tumor regression. This can only be achieved, however, if immunity to the tumor is mediated exclusively by Lyt-2+ T cells, as is the case for the L5178Y lymphoma. In the case of the P815 mastocytoma, treatment with anti L3T4 mAb was without a therapeutic effect, and this was in keeping with the finding that immunity to this tumor is mediated by L3T4+, as well by Lyt-2+ T cells. PMID- 2974067 TI - Specific lysis of allogeneic cells after activation of CD3- lymphocytes in mixed lymphocyte culture. AB - Human CD3- lymphocyte populations were obtained by treating peripheral blood lymphocytes with mAbs directed to CD3, CD4, and CD8 surface antigens. The resulting populations were cultured with irradiated allogeneic cells; at day 4, 100 U/ml IL-2 were added and cultures continued for an additional 10 d. The resulting populations were CD3-CD2+CD7+ and displayed cytolytic activity against PHA-induced blast cells bearing the stimulating alloantigens but not against autologous or unrelated allogeneic blast cells. When CD3- populations were cultured with irradiated autologous cells, no cytolytic activity could be detected either against autologous or allogeneic blast cells. On the other hand, K562 target cells were lysed by both MLC-derived CD3- cell populations regardless of the origin (autologous or allogeneic) of the stimulating cells. CD3- clones were further derived from MLC-stimulated CD3- populations. These clones displayed a cytolytic pattern similar to the original MLC populations as only specific PHA blasts could be lysed. These clones did not express detectable surface TCR alpha/beta or -gamma/delta molecules and lacked productive mRNA for TCR alpha and beta chains, while small amounts of TCR-gamma mRNA were detectable in one of four clones tested. Also mRNA for CD3 gamma and delta chains were undetectable in all clones, however, CD3 epsilon mRNA was consistently present. PMID- 2974068 TI - Immunological properties of the primer-independent glucosyltransferase of Streptococcus mutans serotypes d and g. AB - Streptococcus mutans serotype g secretes at least three kinds of glucosyltransferase with different enzymological and immunological properties. One of them is a primer-independent enzyme and seems to be the source of primer for the others, both of which are primer-dependent enzymes. Recently, we purified the primer-independent enzyme, the third glucosyltransferase in this group from S. mutans strain AHT-k serotype g. In the present study, we examined the specificity of the antiserum against the primer-independent glucosyltransferase using extracellular culture-conditioned fluids of many strains of the various serotypes of S. mutans. The antiserum cross-reacted with the extracellular culture fluids from strains of serotypes d and a, in addition to serotype g, but not with those of other serotypes, indicating that the primer-independent glucosyltransferase is secreted by the S. sobrinus and S. cricetus, but not by S. mutans and S. rattus. The antiserum did not completely inhibit the activity of the enzyme, even at more than twofold antibody excess, determined by indirect precipitation with immobilized staphylococcal protein A. PMID- 2974066 TI - Specific inhibition of lymphokine biosynthesis and autocrine growth using antisense oligonucleotides in Th1 and Th2 helper T cell clones. AB - T helper cells have recently been divided into two subsets. The Th1 subset secretes and responds to IL-2 in an autocrine manner. The Th2 subset upon mitogen or antigen stimulation releases IL-4. Here we describe a novel technology that allowed us to confirm this distinction. We have used synthetic oligonucleotides complementary to the 5' end of mouse IL-2 and IL-4 to specifically block the biosynthesis of IL-2 or IL-4 in two murine helper T cell clones from the Th1 or Th2 subset. We show that the antisense IL-2 oligonucleotide inhibited the proliferation of the Th1 clone and had no effect on the Th2 clone. In parallel experiments, the antisense IL-4 oligonucleotide blocked the proliferation of the Th2 clone and not the proliferation of the Th1 clone. The inhibition was significantly reversed in both cases by the addition of the relevant lymphokine (IL-2 in the case of the Th1 clone, IL-4 in the case of the Th2 clone). Northern analysis, using cDNA probes specific for the two lymphokines, showed a decrease in the steady-state level of the relevant lymphokine mRNA, suggesting the specific degradation of the mRNA by an RNase H-like enzymatic activity. This strategy, which allows the specific blockade of the biosynthesis of a lymphokine, could be useful for future studies on the role of each T helper subset in physiological immune responses. PMID- 2974069 TI - Characterization of B278, a phage different from Mu that also produces auxotrophic mutations in Escherichia coli K12. AB - Bacteriophage B278 has been characterized and compared with Mu, the only phage known to produce random mutations in E. coli. Although both phages are morphologically indistinguishable and have a similar host range, they clearly differ at both the protein and the DNA level. B278 apparently possesses a DNA protection mechanism that is different from the mom system described for Mu. PMID- 2974070 TI - Changes in cortical 5-HT2 receptors following 5-HTP and paroxetine administration in rats. PMID- 2974071 TI - A comparison of statistical outcome variables in extremely low birth weight infants. AB - Three different summary statistics of developmental outcome are described. These include the "handicap ratio" (handicapped individuals divided by survivors), the "handicap rate" (handicapped individuals divided by live births for that birth weight group), and the "handicaps per 10,000 live births" (handicapped individuals divided by 10,000 live births for a geographic region). An analysis of these statistics was undertaken using a sample of extremely low birth weight (ELBW) (less than 1,001 g) infants born in Hawaii during the period of 1975-1981. The results of this study showed a poor correlation between these summary statistics. The results suggest that these statistics represent different aspects of outcome in the ELBW infant. Suitable uses for each of these variables is discussed. PMID- 2974072 TI - Does it matter which cells are infected by the human immunodeficiency virus type 1? PMID- 2974073 TI - The antitumour effect of lipophilic derivatives of 5-fluoro-2'-deoxyuridine incorporated into liposomes. AB - Lipophilic prodrugs of 5-fluoro-2'-deoxyuridine (FUdR), namely 5'-O-palmitoyl-5 fluoro-2'-deoxyuridine (5'-O-palm-FUdR) and 3',5'-O-dipalmitoyl-5-fluoro-2' deoxyuridine (3',5'-O-dipalm-FUdR), were incorporated into bilayer liposomes. Prodrug incorporation into positively charged liposomes was quantitative and stable, homogeneous bilayer vesicles were obtained. The maximal amounts of prodrug incorporation are 200 micrograms for 5'-O-palm-FUdR and 90 micrograms for 3',5'-O-dipalm-FUdR per mg egg phosphatidylcholine as matrix lipid. The prodrug liposome preparations were tested in vivo against mammary carcinoma 13/C, Lewis lung carcinoma and L1210 leukaemia and compared to the cytostatic activity of free FUdR and of the prodrugs dissolved in peanut oil. Intraperitoneally administered prodrugs either incorporated into liposomes or dissolved in peanut oil inhibited tumour growth in all animals. The comparison of the doses required for tumour growth inhibition showed that both prodrugs were active at concentrations 20-75 times lower as compared to unmodified FUdR. However, due to the increased toxicity of the prodrug-liposome preparations, the therapeutic index of the parent drug FUdR could not be improved. The cytostatic effect of the prodrug preparations may be explained by altered pharmacokinetic properties of the FUdR derivatives and the additional sustained release action the liposomes are providing. A further increase of the antitumour activity may be obtained by the attachment of tumour-specific antibodies to the surface of such prodrug containing liposomes. PMID- 2974075 TI - The effect of liposome-entrapped specific cell growth inhibitor (chalone) on tumour cell proliferation. AB - The effect of specific cell growth inhibitor (chalone) on the mitotic activity of ascites ISM applied entrapped in liposomes compared to the non-entrapped 'free' form was investigated. The 'free' chalone injected intraperitoneally in BALB/c ascites ISM-bearing mice (1500 mg per kg body weight) decreased the mitotic activity of the tumour by 66 per cent 2.5 h after administration. Five hours after administration, complete restoration of the mitotic index to the control level was observed. Chalone encapsulated into Bangham liposomes (1000 mg per kg body weight) in otherwise identical conditions produced 44 per cent inhibition after 2.5 h, while at 5 h post-injection the mitotic inhibition was still present increasing up to as much as 50 per cent. No such effect was evident by chalone encapsulated in freeze-thawed liposomes (500 mg per kg body weight). The chalone under study specifically inhibits the cell division of the originating tumour cells and has no effect on its DNA synthesis or on the mitosis of the basal layer of the oesophagus and crypts of the intestinal epithelium. The results obtained provide evidence that liposome-entrapped chalone prolongs the inhibition of tumour cell mitosis in comparison to the 'free' one even in a reduced dose. Some possible applications of this approach concerning cancer research and treatment are discussed. PMID- 2974074 TI - Microcapsules for arterial chemoembolization: appearance and in vitro drug release characteristics. AB - Microcapsules made of eight different capsular materials and containing floxuridine (FUDR) were formulated and evaluated with regard to physical properties (size and structure) and in vitro drug release characteristics. Capsules averaged 100, 150 and 200 micron in diameter and appeared as either round or 'crystalline' particles. The degree and pattern of drug release was dependent on the capsular material used. Alteration in the molecular architecture of a specific material produced changes in the percentage of total encapsulated drug released per unit of time, but not in the pattern of release. Changing capsular size or amount of drug contained within the capsules failed to alter either the pattern of drug release or the percentage of encapsulated drug released per unit of time. However, changes in either parameter did affect the total amount of drug released per unit of time. PMID- 2974076 TI - Advances in prosthetic heart valves: fluid mechanics of aortic valve designs. AB - The in vitro hemodynamic characteristics of a variety of mechanical and tissue heart valve designs used during the past two decades were investigated in the aortic position under pulsatile flow conditions. The following valve designs were studied: Starr-Edwards ball and cage (model 1260), Bjork-Shiley tilting disc (convexo-concave model), Medtronic-Hall tilting disc, St. Jude Medical bileaflet, Carpentier-Edwards porcine and pericardial (models 2625, 2650 and 2900), Hancock porcine (models 250 and 410) and Ionescu-Shiley standard pericardial. The Starr Edward ball and cage, Bjork-Shiley tilting disc, Carpentier-Edwards porcine (model 2625) and Ionescu-Shiley standard pericardial valves were designed prior to 1975, while the Medtronic-Hall tilting disc, St. Jude Medical bileaflet, Hancock porcine (model 250), Hancock II porcine (model 410), Carpentier-Edwards porcine (model 2650) and Carpentier-Edwards pericardial (model 2900) valves were designed after 1975. The pressure drop results indicated that the valves designed prior to 1975 had performance indices of 0.30 to 0.45, whereas the valves designed after 1975 had performance indices of 0.40 to 0.70. The regurgitant volumes were higher for the mechanical designs (5.0 to 11.0 cm3/beat) compared to the tissue bioprostheses (1.0 to 5.0 cm3/beat). Two-dimensional laser Doppler anemometry studies indicated that the valves designed after 1975 tended to create more centralized flow fields, with reduced levels of turbulent shear stresses. However, none of the current valve designs is ideal: they all create areas of stasis and/or regions of low velocity reverse flow; and regions of elevated turbulent shear stresses that are capable of causing sub-lethal and/or lethal damage to the formed elements of blood. PMID- 2974077 TI - Impairment of T-helper function by a Plasmodium berghei-derived immunosuppressive factor. AB - Mice injected with an immunosuppressive factor (ISF) extracted from Plasmodium berghei-infected rat erythrocytes have a reduced antibody response to unrelated antigens. T-cells from ISF-treated mice failed to provide adequate help to naive, syngeneic B-cells in the primary IgM response in vitro to sheep red blood cells and to dinitrophenylated keyhole limpet hemocyanin. The same T-cells, however, were able to cooperate with memory B-cells in the secondary IgG response. No other cellular deficit was detected in ISF-treated mice; B-cells and macrophages behaved normally, and there was no detectable excess of suppressor cells. The T cell impairment was not reflected in decreased production of interleukin 2, but was also shown by the diminished delayed type hypersensitivity reaction to sheep red blood cells of ISF-treated mice. PMID- 2974078 TI - Organization of the Euplotes crassus micronuclear genome. AB - Euplotes crassus, like other hypotrichous ciliated protozoa, eliminates most of its micronuclear chromosomal DNA in the process of forming the small linear DNA molecules that comprise the macronuclear genome. By characterizing randomly selected lambda phage clones of E. crassus micronuclear DNA, we have determined the distribution of repetitive and unique sequences and the arrangement of macronuclear genes relative to eliminated DNA. This allows us to compare the E. crassus micronuclear genome organization to that of another distantly related hypotrichous ciliate, Oxytricha nova. The clones from E. crassus segregate into three prevalent classes: those containing primarily eliminated repetitive DNA (Class I); those containing macronuclear genes in addition to repetitive sequences (Class II); and those containing only eliminated unique sequence DNA (Class III). All of the repetitive sequences in these clones belong to the same highly abundant repetitive element family. Our results demonstrate that the sequence organization of the E. crassus and O. nova micronuclear genomes is related in that the macronuclear genes are clustered together in the micronuclear genome and the eliminated unique sequences occur in long stretches that are uninterrupted by repetitive sequences. In both organisms a single repetitive element family comprises the majority of the eliminated interspersed middle repetitive DNA and appears to be preferentially associated with the macronuclear sequence clusters. The similarities in the sequence organization in these two organisms suggest that clustering of macronuclear genes plays a role in the chromosome fragmentation process. PMID- 2974079 TI - [Endoluminal angioplasty of iliac obstruction. Immediate results and cumulative patency at 2 years in 23 patients]. AB - 23 patients with obstruction of one or both main iliac arteries underwent one or more transluminal angioplasties by catheterization via the obstructed vessel. 26 iliac arteries were thus reopened. The length of obstructions varied from 1 to 10 cm. The majority of the obstructions were of recent onset whilst in 6 cases the presumed date was more than 6 months previously. In all cases potency was obtained with normal follow-up angiograms in 20 cases, and a residual though not hemodynamically significant stenosis in 6 cases. All patients were followed up by Doppler and in the majority of cases by venous digital angiography, with a follow up at the present time of 6 months to 2 years. 2 patients underwent surgery because of reobstruction occurring 2 and 13 months after dilatation. Cumulative patency is 79% in 2 years, similar to results obtained by surgery. These results show that iliac angioplasty, a simple procedure, may avoid or delay surgery, and that it should be widely used in this precise anatomical indication. PMID- 2974080 TI - [Isolated posterior pre-marginal hernia. Apropos of 6 cases]. AB - Posterior pre-marginal disk hernia, a rare lesion, was diagnosed in six young adults with lumbago. Lateral lumbar spine radiographic images showed an unusual appearance, combining a posterior marginal defect of a vertebral corner and a small bony spicule projecting into lumbar canal. A literature review revealed 27 similar cases, mostly in adolescents. These vertebral disk anomalies are considered by some authors to be fractures of the posterior margin border. A more plausible hypothesis is a pre-marginal hernia, because of lack of a history of injury, the radiologic appearance comparable with that of sequelae of Scheuermann's disease, and the frequent association of these lesions with a spinal growth dystrophy. PMID- 2974081 TI - Similarity of vasopressin receptors in seminal vesicles and renal medulla of pigs. AB - To test the hypothesis that the vasopressin receptors found in seminal vesicles are similar to those present in the renal tubules competition experiments were performed with vasopressin and several analogues with different specificities for the V1 and V2 subtypes of vasopressin receptor. Autoradiographic studies were carried out on sections from seminal vesicles and kidney to identify the cellular target of vasopressin. Vasopressin receptors in renal medulla and seminal vesicles of pigs shared the same rank order of potency for vasopressin and its analogues and were localized in the epithelium of the seminal vesicles and in collecting tubules of renal medulla. These results strongly suggest that the vasopressin receptors present in kidney and seminal vesicles belong to the same subtype, V2, of vasopressin receptor. PMID- 2974082 TI - Periarticular fibrosis associated with idiopathic retroperitoneal fibrosis. AB - We describe a previously unreported association of retroperitoneal fibrosis with biopsy proven periarticular inflammatory fibrosis. Histologic features were strikingly similar at the 2 sites. Plasma levels of a platelet derived growth factor, connective tissue activating peptide III, were found to be elevated and may play a role in pathogenesis. A dramatic response of the periarticular process to corticosteroids correlated with improvement of other variables of disease activity. PMID- 2974083 TI - Introducing child survival and development topics into the medical curriculum of a developing country. PMID- 2974084 TI - Stage transitioning in the sensorimotor development of Down's syndrome infants. AB - The relative amount of time necessary to move from stage-to-stage during the sensorimotor period was examined among a group of 30 Down's syndrome and 12 non retarded infants. Stage transitioning was examined for the four transitional points between Stages II and VI for the seven branches of sensorimotor development on the Uzgiris-Hunt scales. The findings showed that the Down's syndrome infants generally took longer to move from one stage to the next even after their slower pace of development was taken into consideration. PMID- 2974085 TI - Structure and organization of Marchantia polymorpha chloroplast genome. II. Gene organization of the large single copy region from rps'12 to atpB. AB - The nucleotide sequence (56,410 base-pairs) of the large single-copy region of chloroplast DNA from the liverwort Marchantia polymorpha has been determined. The sequence starts from one end (JLA) of the large single-copy region and encompasses genes for 21 tRNAs, six ATPase subunits (atpA, atpB, atpE, atpF, atpH and atpI), two photosystem I polypeptides (psaA and psaB), four photosystem II polypeptides (psbA, psbC, psbD and psbG), five ribosomal proteins (rps2, rps4, rps7, rps'12 and rps14), and three RNA polymerase subunits (rpoB, rpoC1 and rpoC2). In addition, we detected 18 open reading frames ranging from 29 to 2136 amino acid residues long, four of which share significant amino acid sequence homology to those of an Escherichia coli malK protein (designated mbpX), human mitochondrial ND2 (ndh2) and ND3 (ndh3) of a respiratory chain NADH dehydrogenase, or a bacterial antenna protein of a light-harvesting complex (lhcA). Sequence analysis suggests that four tRNA genes and six protein genes might be split by introns; they are trnG(UCC), trnK(UUU), trnL(UAA), trnV(UAC), atpF, ndh2, rpoC1, rps'12, ORF135 and ORF167. In the large single-copy region described here, the gene organization deduced is highly conserved with respect to that of higher plants, but an inversion of some 30,000 base-pairs flanked by trnL(CAA) and trnD(GUC) was seen between the liverwort and tobacco chloroplast genomes. PMID- 2974086 TI - Crystallization and preliminary X-ray studies of T4 phage beta glucosyltransferase. AB - Crystals of the DNA glucosylating enzyme beta-glucosyltransferase from phage T4 have been grown in the presence of uridine diphosphate glucose. The crystals are orthorhombic, space group P2(1)2(1)2 with a = 148.3 A, b = 52.6 A, c = 52.6 A. The assumption of one monomer of Mr 40,000 per asymmetric unit gives rise to a Vm of 2.56 A 3/dalton. The crystals diffract to beyond 2.7 A and are suitable for X ray structure analysis. PMID- 2974088 TI - Mechanism and significance of abnormal blood level of beta 2-microglobulin in patients with Graves' disease. PMID- 2974089 TI - [Development of new hypoxic cell radiosensitizer in Japan]. AB - Basic studies in hypoxic cell radiosensitizers developed in Japan were reviewed. Because of the unsuccessful clinical trials of misonidazole (MISO), many efforts have been made to find a new hypoxic cell sensitizer which is more effective and/or less toxic than MISO. Already over a thousand drugs were tested, but the majority of them did not work in vivo or were very toxic to the animals. Finally four drugs, KU-2285, KIH-801 (802), RK-28 and RP-170, have been proven to be effective both in vitro and in vivo. All of them are derivatives of 2 nitroimidazole. The first two have different side chains of fluorinated amide and acetohydroxamic acid, respectively. The second two both have sugar moieties. Investigations are now under way to find possible clinical applicability. PMID- 2974087 TI - Effect of subchronic dieldrin treatment on calmodulin-regulated Ca2+ pump activity in rat brain. AB - Calmodulin-dependent (CaM-dependent) and nondependent Ca2+ pump activity was determined in the brain P2 fraction of Sprague-Dawley rats treated in vivo with 10 ppm dieldrin for 60 d. After 40 d of dieldrin intoxication, daily food consumption and body weight gain were found to be affected. Dieldrin inhibited both CaM-dependent and -independent Ca2+-ATPase activity and the enzyme activity decreased to 36% (CaM-dependent)--and 18% (basal) respectively after 60 d of intoxication. Reduction in brain CaM levels and a subsequent inhibition of Ca2+ ATPase activity suggests that chronic intoxication of dieldrin at very low doses alters calmodulin and its regulation of Ca2+-ATPase activity. PMID- 2974090 TI - [Ainu midwifery technic practiced by Mrs. Aiko: lower backache during and after labor. I]. PMID- 2974091 TI - Activation of chicken complement by avian viruses. PMID- 2974092 TI - [Characteristics of rate and time parameters in secondary changes in the ventricular repolarization period]. AB - An analysis of the first electrocardiographic derivative in 48 patients with left ventricular hypertrophy and complete block of the left limb of His bundle demonstrated a prolongation of myocardial excitation spread and increased rate of change in the difference of potentials during repolarization, the changes being more marked in patients with left-ventricular hypertrophy. In cases where left ventricular hypertrophy was combined with coronary disease, the rate of change in the difference of potentials was reduced dramatically during repolarization. PMID- 2974093 TI - [Electrocardiographic characteristics of an anomalous origin of the left coronary artery from the pulmonary trunk]. AB - Electrocardiographic characteristics of abnormal left coronary arterial branching (ALCAB) from the pulmonary trunk were examined in 36 patients, aged 2 months to 39 years. Signs of marked left-ventricular and left-atrial overstrain were typical for all ages. There were signs of past left ventricular myocardial infarction in 88.8% of patients, the cicatricial changes being anterolateral in 65.6%, which is typical for the defect. The follow-up of ECG pattern over 1 month to 8 years after the defect was first detected demonstrated that coronary circulation deteriorated in 64.3% of cases. PMID- 2974094 TI - Hypertension in autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) has been shown to be associated with a greater than 50 percent incidence of hypertension prior to deterioration in renal function as assessed by glomerular filtration rate. The present study provides evidence for increased cardiac pre-load, as assessed by plasma atrial natriuretic factor (ANF) and cardiac index, in hypertensive as compared to normotensive ADPKD. The hypertensive ADPKD patients exhibited an increased renal vascular resistance as compared to the normotensive patients in spite of comparable glomerular filtration rates. It is hypothesized that the renal involvement of hypertensive ADPKD patients causes an impaired renal response to the observed increase in cardiac index, and also may release a venoconstrictor (such as angiotensin) which contributes to the enhanced cardiac pre-load and thus the hypertension. PMID- 2974095 TI - Effect of synthetic human atrial natriuretic peptide (102-126) in nephrotic syndrome. AB - Synthetic human ANP (102-126) or vehicle was intravenously administered to eight patients with non-edematous nephrotic syndrome to study its effect on protein and sodium excretion. ANP was given in ascending doses, each dose for one hour, two to three days apart. Four patients received 0.03, 0.10 and 0.45 microgram/kg/min of ANP, and four received 0.015, 0.06 and 0.20 microgram/kg/min. Natriuresis increased at all doses; by 179 +/- 13.6% (mean +/- SEM; P less than 0.05) at 0.015 microgram/kg/min and by 660 +/- 71.5% (P less than 0.01) at 0.20 microgram/kg/min. Urinary albumin excretion increased by 138 +/- 30.1% (P less than 0.05) at 0.015 microgram/kg/min of ANP and by 534 +/- 132% (P less than 0.01) at 0.20 microgram/kg/min. Immunoglobulin G excretion increased proportionally to albumin excretion. Hematocrit and serum albumin concentration increased after ANP. In each patient the percent reduction of plasma volume calculated from the effect on serum albumin was smaller than the hemoconcentration calculated from the effect on hematocrit, suggesting a loss of albumin from the intravascular compartment. This could not be accounted for by the increased glomerular filtration of albumin. Blood pressure and effective renal plasma flow decreased and filtration fraction increased after ANP. Plasma renin was suppressed at lower doses of ANP but was stimulated, together with plasma noradrenaline, at higher doses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974096 TI - Mechanisms of action of vasopressin and vasopressin antagonists. PMID- 2974097 TI - Use of vasopressin antagonists in human diseases. PMID- 2974098 TI - [Communication with patients during respirator therapy]. PMID- 2974099 TI - [Causes, clinical aspects and therapy of acne vulgaris]. PMID- 2974100 TI - Immune complex acute necrotizing glomerulonephritis with progression to diffuse glomerulosclerosis. A murine model. AB - Male BALB/c mice given daily intraperitoneal injections of 4 mg of horse-spleen apoferritin develop, in the majority of cases, a proliferative and necrotizing glomerulonephritis with leukocytic infiltration and extensive intraglomerular thrombosis within 10 to 14 days, as previously reported. We now show that if injection of the antigen is discontinued, surviving animals develop extensive glomerulosclerosis (GS). Ten of 13 mice treated as indicated above and sacrificed 4 months after the last horse-spleen apoferritin injection developed segmental GS involving over 40% of their glomeruli. Tubulointerstitial damage of proportionate severity also developed. Ultrastructurally, pronounced mesangial expansion due to matrix deposition obliterated the glomerular architecture. We offer this as a reproducible model of immune complex-mediated GS particularly suited to the study of cellular interactions involved in the pathogenesis of GS. PMID- 2974101 TI - The regulation of autoantibody production in Heymann's nephritis by T lymphocyte subsets. AB - The role of T lymphocyte subsets in the regulation of the autoantibody response to renal tubular antigen and the development of Heymann's nephritis (HN) in different rat strains was investigated in the present study. WF rats were highly susceptible to HN induction, PVG rats were moderately susceptible, whereas DA and BN were resistant. The induction of an autoantibody response to renal tubular antigen and HN in PVG rats was T cell-dependent as they were abrogated by neonatal thymectomy and T cell depletion after adult thymectomy, whole body irradiation and bone marrow reconstitution. Reconstitution of neonatal thymectomized and adult thymectomized whole body irradiated and bone marrow reconstitution of PVG rats with different T lymphocyte subsets revealed that the induction of an autoantibody response to renal tubular antigen in these animals required help from a long lived, W3/25+ T cell subset and was down regulated by a MRC OX8+ T cell subset. Adoptive transfer experiments demonstrated that the suppressor T cells involved were short lived and were probably generated by the interaction between a W3/25+ inducer and a MRC OX8+ precursor subpopulation. The suppressor precursor T cells were recently derived from the thymus as the suppressor T cell response was abrogated by adult thymectomy, an effect which could only be fully restored by thymic grafting or injection of adult thymocytes but not by the injection of normal spleen or lymph node cells. As adult thymectomy had less effect on the autoantibody response and HN development in the highly susceptible WF rats, the difference in disease susceptibility between the WF and PVG rats may be the result of different suppressor T cell recruitment from the adult thymus. Since adult thymectomy did not affect disease development in the DA and BN rats, the recently thymus-derived suppressor T cells did not contribute to innate disease resistance in these rat strains. PMID- 2974102 TI - Structural remodeling of cardiac myocytes in rats with arteriovenous fistulas. AB - Structural changes in the heart associated with chronic volume overloading have not been adequately documented. In anesthetized adult rats, microsurgical techniques were used to produce two types of arteriovenous shunts. The end of a femoral artery was connected to the side of a femoral vein to produce a femoral shunt. Aortocaval fistulas were produced by placing a venous graft (femoral vein) between the abdominal aorta and inferior vena cava. A polyethylene ring was placed around the venous graft to control the size of the shunt. After 10 weeks, rats were anesthetized and hearts were excised, weighed, and perfused with collagenase to obtain isolated myocytes. Heart weight/body weight of rats increased 15% (p less than 0.005) with unilateral femoral fistulas and 41% (p less than 0.001) with aortocaval fistulas. Length of isolated cardiac myocytes was measured directly with a microscope. Cell volume was measured with a Coulter Channelyzer. Myocyte cross-sectional area was calculated from cell volume/cell length. Cell volume increased in proportion to heart weight in each experimental model. With both types of shunts, the majority of myocyte hypertrophy was due to an increase in cell length. There was a trend toward an increase in cross sectional area in each region of both fistula groups, although this change was not statistically significant. These results indicate that eccentric cardiac hypertrophy was associated with enlargement of individual myocytes primarily as a result of increased length. PMID- 2974103 TI - Androgen levels during sequential insulin euglycemic clamp studies in patients with polycystic ovary disease. AB - It is postulated that insulin may play a role in the regulation of ovarian androgen production. In order to test the possible interrelation between serum insulin levels and androgen production, sequential euglycemic insulin clamp (Mode 9:1 on Biostator, insulin infusion rate: 0.1; 0.2 and 0.4 U/kg b. wt/h, each rate for 90 min, BC = 80 mg/dl) was done in 6 patients with polycystic ovary disease and normal glucose tolerance. Insulin, C-Peptide, testosterone and dehydroepiandrosterone-sulphate were measured in 0, 70, 80, 90, 160, 170, 180, 250, 260 and 270 min. Significant suppression of C-Peptide levels were achieved (0 min vs 270 min = 0.81 + 0.25 vs 0.15 + 0.20 nmol/l; P less than 0.05). Basal insulin as well as the mean plateau for each insulin infusion rate were as follows: 28 + 9; 248 + 119; 427 + 69 and 524 + 77 microU/l. There was significant testosterone increase at the end of insulin infusion (0 vs 270 min = 4.8 + 1.2 vs 8.1 + 1.7 nmol/l; P less than 0.05). There were no significant changes in dehydroepiandrosterone-sulphate levels during clamp studies (0 vs 270 min = 1055 + 133 vs 913 + 114 ng/ml; P greater than 0.05). It is concluded that acute insulin infusion under the condition of sequential euglycemic clamp could increase androgen production in the ovaries of patients with PCO. PMID- 2974104 TI - Heparan sulfate is necessary for adhesive interactions between human early hemopoietic progenitor cells and the extracellular matrix of the marrow microenvironment. AB - Human hemopoietic blast colony-forming cells (BI-CFCs) recognize and adhere to the extracellular matrix (ECM) produced by marrow-derived stromal cells in vitro. We have investigated the requirements for this interaction by testing the capacity of BI-CFCs to adhere to ECM components under a variety of conditions. Binding was prevented completely by prior treatment of stromal ECM with nitrous acid, in large part by treatment with heparitinase or hyaluronidase, and slightly by treatment with chondroitinases. Whereas heparan sulfate isolated from marrow stromal cultures effectively blocked binding, heparan sulfate from bovine kidney did not. Chondroitin sulfate and hyaluronic acid did not have any effect in this test. In contrast, collagen was not sufficient for the interaction because dishes coated with collagen type I or IV did not act as adhesive surfaces for BI-CFCs. Ligands for integrin receptors (e.g., fibronectin) did not participate in BI-CFC binding because the synthetic pentapeptide glycine-arginine-glycine-asparagine serine did not compete with stroma in binding BI-CFCs. These findings indicate that heparan sulfate in the bone marrow microenvironment is necessary for BI-CFC binding to ECM and may contribute to localizing hemopoietic stem cells in hemopoietic tissue. PMID- 2974105 TI - Early involvement of the fim-2 and fim-3 regions in mouse myeloblastic leukemogenesis. AB - Retroviruses lacking oncogenes induce tumors or leukemias after a long latency which generally exceeds several months. Cellular transformation most probably results from the activation of cellular oncogenes or putative proto-oncogenes due to proviral integration. Several genetic changes are likely to be necessary for the appearance of fully malignant cells. However, the sequence of genetic changes initiating and leading to malignant transformation is difficult to study since, in most experimental conditions, the only accessible cells are fully transformed cells. We have previously described an in vitro model of murine myeloblastic leukemogenesis during which several successive steps leading to fully malignant and transplantable cells have been identified. This in vitro transformation process develops over approximately a 1-year period. In this paper, we demonstrate that frequent cellular DNA rearrangements due to proviral integrations in specific regions occur early in the myeloblastic transformation process and remain stable throughout the in vitro leukemogenesis, and in tumors derived from in vitro fully transformed myeloblasts. PMID- 2974106 TI - Control of in vivo differentiation of myeloid leukemic cells. AB - The differentiation of leukemic cells in vivo can be a useful approach to therapy. In vivo differentiation of myeloid leukemic cells was studied in intraperitoneally implanted diffusion chambers, containing different soluble antigens. The presence of these antigens in the chambers induced differentiation of myeloid leukemic cells and this was inhibited in immune-deficient mice. Transfer of normal spleen cells enriched for T-lymphocytes or antigen-specific helper T lymphocyte cell lines to mice in which differentiation of leukemic cells was inhibited, restored in vivo differentiation of the leukemic cells. Antigen specific helper T cells produce myeloid regulatory proteins and can accumulate at a site that contains the specific antigen. It is suggested that migration in response to antigen of helper T cells producing regulatory proteins may play an important role in inducing in vivo differentiation of leukemic cells. We have identified a class of myeloid leukemic cells that can be induced to differentiate in vitro by incubation with pure MGI-1GM (GM-CSF) or IL-3, but not with MGI-1G (G CSF). Experiments with pure recombinant proteins have shown that MGI-1GM and IL 3, but not MGI-1G, can also induce these myeloid leukemic cells to differentiate in vivo. These results and our previous studies on the myeloid cell differentiation-inducing protein MGI-2, demonstrate the potential use of normal hematopoietic regulatory proteins not only in regulation of normal hematopoiesis, but also in the treatment of myeloid leukemia by in vivo induction of terminal cell differentiation. PMID- 2974107 TI - The Oxford Optical Pointer: a direction-sensing device with proportional electrical output. PMID- 2974109 TI - [Lichenoid pityriasis. Immunologic study of 10 children]. AB - Ten children clinically and histologically diagnosed as having pityriasis lichenoides (PL), have been studied by direct immunofluorescence (DIF). Circulating immune complexes (CI) have also been studied in four children. Granular deposits of IgM, located in the walls of the dermal vessels have been observed in two cases, but they have never been found at the dermo-epidermal junction. Granular deposits of C3 have been observed in three children, both in the walls of the dermal vessels and at the dermo-epidermal junction. The search for immune complexes gave negative results in all cases. The hypothesis of some authors that PL is an immune complex disease cannot be confirmed by our findings. PMID- 2974108 TI - [Progressive symmetric erythrokeratodermia. Report of a case with delayed onset treated by etretinate]. AB - A case of delayed onset (17 years) progressive symmetric erythrokeratodermia is presented. The treatment with etretinate initially improved the clinical picture, but it had to be discontinued because of personality changes, particularly depressive crisis. In addition, clinical manifestations, histology, kinetics and ultrastructural changes in this condition are also commented on. PMID- 2974110 TI - [Adjustment to illness--strategies and goals of health care and outside health care]. PMID- 2974111 TI - [Backache--a condition difficult to treat]. PMID- 2974112 TI - [Psychosocial factors concerning the occupational environment are often causes of low back pain]. PMID- 2974113 TI - Neurotization of pectoralis major myocutaneous flap by the hypoglossal nerve in tongue reconstruction: clinical and experimental observations. AB - Recently, neurotization has been proposed for providing mobility to the pectoralis major, or other myocutaneous flap in lingual reconstruction following total glossectomy. The development of an active tongue-like structure may offer the patient higher potential for rehabilitation of speech and deglutition. The purpose of this thesis is to report experimental and clinical observations on neurotization of the pectoralis major myocutaneous flap. The pectoralis major myoflap of 16 rats was reinnervated by either a hypoglossal nerve pedicle of hypoglossal-genioglossus muscle neuromuscular pedicle. Functional flap reinnervation was confirmed in eight of the 16 animals. The author's clinical experience with reconstruction of the tongue utilizing neurotized pectoralis major myocutaneous flap is presented. PMID- 2974114 TI - [The heart--a gland with endocrine effects]. PMID- 2974116 TI - [Evaluation of water-electrolyte metabolism in patients with upper GI tract cancer--the dynamic status of secretion of alpha-hANP levels]. AB - For the purpose of evaluation of the water-electrolyte metabolism in upper GI tract surgery, I measured alpha-human atrial natriuretic polypeptide (alpha-hANP) and other parameters of twenty-eight patients daily for seven days after operation. The subjects were divided in four groups as follows; 6 r-thoracal and abdominal esophagectomy (r-TA group); 4 l-thoracoabdominal total gastrectomy (1 TA group), 8 abdominal total gastrectomy (TMR group) and 10 partial gastrectomy (PMR group). Plasma alpha hANP levels in r-TA group were significantly higher than those in PMR group on the first, second, third and seventh postoperative days (p less than 0.05). Furthermore, those in 1-TA group were also significantly higher compared to those in PMR group on the second and seventh postoperative days (p less than 0.05). Significant correlation between plasma alpha hANP level and water balance was detected in transabdominal groups (TMR, PMR). However, this correlation was not found in thoracoabdominal groups (r-TA, l-TA). There was no correlation among alpha hANP level, heart rate and blood pressure. Serum and urinary Na/K ratio was lower in r-TA group, l-TA group, TMR group, and PMR group in descending order. Consequently, plasma alpha hANP level plays an important role in water-electrolyte metabolism during the perioperative period. A good group of the dynamic status of the secretion of alpha hANP levels might be a very useful index for the evaluation of the magnitude of surgical stress. PMID- 2974115 TI - [The Arys-MT barocomplex]. PMID- 2974117 TI - [Facilitation of tumor metastasis to the lung by operative stress in the rat- influence of adrenocortical hormones and preoperative administration of OK-432]. AB - Experimental studies were performed to clarify the mechanism of facilitation of tumor metastasis to the lung by operative stress, using an experimental model in which 5-week old female Sprague-Dawley (SD) rats were inoculated with a low antigenic and easily metastasizable tumor, MRMT-1. In particular, relevance of adrenocortical hormones to the facilitation of tumor metastasis was examined. Furthermore preventive effects of preoperative administration of a nonspecific immunopotentiator, OK-432, were examined. Number of metastatic nodules was increased significantly by operative stress and the increase was proportionate to severity of the stress. The facilitation of metastasis by operative stress was significantly inhibited by preoperative OK-432 administration. In adrenalectomized rats, no such facilitation of metastasis by operative stress was observed. After administration of 2.5 to 20mg of hydrocortisone, number of metastatic nodules increased dose-dependently. The increase in metastatic nodules by administration of 5mg of hydrocortisone was inhibited by preoperative OK-432 administration. Thus it was concluded that facilitation of tumor metastasis by operative stress was proportionate to severity of the stress, and one of its essential causative factors was the stress-induced adrenocortical hypersecretion, which suppressed immunity of the host. Administration of OK-432 was effective for counteracting the stress-induced facilitation of metastasis. PMID- 2974118 TI - [Influence of flow disturbance on an anastomotic intimal hyperplasia: experimental and clinical study]. AB - Experimental and clinical studies were made of the localization and morphology of anastomotic intimal hyperplasia (AIH) at the end-to-side anastomosis in relation to flow disturbances. In vitro experimental findings showed that boundary layer separation (BLS) became prominent as proximal outflow segment (POS) flow increased. An aorto-right iliac bypass was performed on 30 dogs using 22 Biografts, 3 EPTFES and 5 Dacron grafts. Distal end-to-side anastomoses were made at 3 different angles, i.e., 30 degrees (Group I), 90 degrees (Group II) and 150 degrees (Group III). BLS was likely to occur at the toe in Group I because POS flow exceeded 50%, while not in Group III. Up to 35 months observation, AIH was noted to develop at the toe 36% in Group I, 25% in Group II but in none in Group III. Thirty five reconstructions using Biograft in which angiography was performed later than 6 months postoperatively were chosen for clinical study. Invariably in all cases of AIH occurring following a femoropopliteal bypass, severe stenosis was noted to occur at the toe and heel of the distal end-to-side anastomosis. In conclusion, a disproportionate increase in POS flow was considered a likely cause of marked BLS and, consequently, an important precipitating cause of AIH. When POS flow exceeds 50%, a distal anastomosis dividing flow distribution should be considered as a salvage operation of potential usefulness. PMID- 2974119 TI - [Healing of an external velour warp-knit Dacron vascular prosthesis]. AB - An external velour warp-knit Dacron vascular prosthesis (8mm in internal diameter, 6cm in length), whose porosity is 1200cc/cm2/min at 120mmHg, was implanted in the descending thoracic aorta of 30 mongrel dogs. The dogs were sacrificed at different intervals between 2 weeks and 6 months. All grafts were patent at sacrifice and no serious complication was observed. Thrombus free surface score was about 90%. Cell coverage on the flow surface was first observed near anastomoses in 1-month specimens and was almost completed in 2 months. Transinterstices healing was completed in 2 months. This excellent healing in a canine model would allow clinical availability of this graft. PMID- 2974120 TI - The effects of pertussis toxin on autoreceptor-mediated inhibition of dopamine synthesis in the rat striatum. AB - Activation of synthesis-modulating dopamine autoreceptors by dopamine or its agonists has been shown to inhibit dopamine synthesis in the rat striatum. However, systemic administration of the direct-acting dopamine agonist apomorphine failed to inhibit dopamine synthesis in striata from rats that had received local unilateral administration of pertussis toxin. Apomorphine did reduce dopamine synthesis by greater than 50% in sham injected control rats as well as in the striata opposite to the side of pertussis toxin injection. Examination of G proteins in striatal tissue revealed that 61% of the G proteins were ADP-ribosylated in vivo by direct pertussis toxin injection. These data suggest that guanine nucleotide regulatory proteins mediate the effects of activation of striatal synthesis-modulating dopamine autoreceptors. PMID- 2974121 TI - Platelet function in type I diabetes mellitus. PMID- 2974122 TI - The role of the leucine zipper in the fos-jun interaction. AB - Mutagenesis of the fos protein supports the hypothesis that a heptad repeat of leucine residues stabilizes the interaction between the fos and jun proteins. We show that the complex between fos and jun can bind to DNA more tightly than either protein alone and that basic residues adjacent to the leucine repeat of fos contribute to the DNA-binding potential of the complex. PMID- 2974124 TI - Recovery from mild concussion. PMID- 2974123 TI - Emotional adjustment in compensated head injury patients. AB - Patterns of emotional adjustment based on the Minnesota Multiphasic Personality Inventory (MMPI) were examined in a group of 124 patients who had suffered head injuries, reported postconcussional symptoms, and were receiving compensation. Cluster analytical procedures were used to identify subgroups. The sample was divided into two equivalent groups for purposes of crossvalidation. The results of the analyses in the two samples yielded similar patterns. Within each subgroup, the groups differed on the level and pattern of emotional disturbance. Approximately 65% of the sample had normal profiles or only mild elevations on the "neurotic" scales. The largest subgroup had mild elevations. After the exclusion of invalid profiles, a small proportion had severe disturbances. The injury-related neurological characteristics of these groups are similar. These findings are discussed in terms of the previous literature on the effects of compensation on posttraumatic personality adjustment. PMID- 2974125 TI - Quantitative morphometric analysis of two types of serotonin-immunoreactive nerve fibres differentially responding to p-chlorophenylalanine treatment in the rat brain. AB - Two types of serotonin neurons differentially responding to treatment with p chlorophenylalanine, a potent inhibitor of serotonin synthesis, have been demonstrated in the rat brain by immunohistochemistry using a serotonin monoclonal antibody in combination with a quantitative morphometric analysis. In control rats, the numerical density of serotonin-immunoreactive varicosities and terminals varied considerably from region to region. In p-chlorophenylalanine treated rats, the numerical density was reduced in every region, but the degree of reduction in each region differed markedly. No correlation was seen in values of the numerical density between the control and p-chlorophenylalanine-treated rats, indicating that serotonin-positive axons in p-chlorophenylalanine-treated rats were present independent of the degrees of the axon density seen in the controls. It is postulated, therefore, that there are at least two types of positive axons; one highly sensitive to p-chlorophenylalanine treatment and another relatively resistant to it. Since the numerical density in any region was always smaller in p-chlorophenylalanine-treated rats than in the controls, p chlorophenylalanine-sensitive axons appeared to be more or less distributed in every region where positive axons were detectable under normal conditions. However, a number of regions were innervated purely by p-chlorophenylalanine sensitive axons. By contrast, p-chlorophenylalanine-insensitive axons were observed in several definite regions. These regions appeared to be related to the limbic system, or cranial motor and parasympathetic nuclei. In the latter nuclei, p-chlorophenylalanine-insensitive axons seemed to possess such common morphological features as having relatively large varicosities and being situated adjacent to neural somata, suggesting their inhibitory roles in neurotransmission. The present result indicates that the two types of serotonin axons may play important roles in the behavioural events observed in rats following p-chlorophenylalanine treatment. PMID- 2974126 TI - 5-Hydroxytryptamine-immunoreactive neurons and nerve fibers in the superior cervical ganglion of the rat. AB - Indirect immunofluorescence method was used to localize 5-hydroxytryptamine immunoreactive structures in the superior cervical ganglion of adult rats. In the ganglia of normal rats, 5-hydroxytryptamine immunoreactivity was localized in the small intensely fluorescent cells, but not in principal nerve cells. In the superior cervical ganglion of rats, pretreated with nialamide, a monoamine oxidase inhibitor, 150 mg/kg i.p., and the 5-hydroxytryptamine precursor, L tryptophan, 45 mg/kg i.p., a large number of 5-hydroxytryptamine-immunoreactive principal nerve cells and small intensely fluorescent cells were detected. The immunoreactive principal nerve cells had long processes, and 5-hydroxytryptamine immunoreactive nerve fibers were observed traversing the ganglion. In ganglia of rats pretreated with colchicine, occasional 5-hydroxytryptamine-immunoreactive principal nerve cells and several small intensely fluorescent cells were detected. Ligation of the main postganglionic nerve trunks of the superior cervical ganglion of normal rats resulted in the appearance of several 5 hydroxytryptamine-immunoreactive principal nerve cells and nerve fibers in the ganglion. To study whether the 5-hydroxytryptamine immunoreactivity in the superior cervical ganglion represented uptake or synthesis of 5 hydroxytryptamine, rats were injected with a specific 5-hydroxytryptamine uptake inhibitor, fluoxetine, 10 mg/kg i.p. twice a day for 5 days, and then they were treated with nialamide and L-tryptophan, as described above. In the superior cervical ganglion of fluoxetine-treated rats, a few 5-hydroxytryptamine immunoreactive principal nerve and small intensely fluorescent cells, as well as some nerve fibers, were detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974128 TI - Management of low back pain in the military population. PMID- 2974127 TI - Localization of quinolinic acid metabolizing enzymes in the rat brain. Immunohistochemical studies using antibodies to 3-hydroxyanthranilic acid oxygenase and quinolinic acid phosphoribosyltransferase. AB - Specific antibodies raised in rabbits against 3-hydroxyanthranilic acid oxygenase (EC 1.13.11.6) and quinolinic acid phosphoribosyltransferase (EC 1.13.11.6) and quinolinic acid phosphoribosyltransferase (EC 2.4.2.19) were used in immunohistochemical studies to map the cellular localization of the quinolinic acid metabolizing enzymes in the adult male rat brain. 3-Hydroxyanthranilic acid oxygenase immunoreactivity was found to be present in glial cells of presumed astroglial identity, as judged by co-localization with glial fibrillary acidic protein. 3-Hydroxyanthranilic acid oxygenase-immunoreactive glial cells were present in all brain regions and within major fiber tracts. The density of 3 hydroxyanthranilic acid oxygenase-immunoreactive glial cells as well as the intensity of staining of these cells differed among brain regions. In general, telencephalic acid diencephalic areas harbored a larger number of 3 hydroxyanthranilic acid oxygenase-positive cells than did mesencephalic regions. In the former regions the caudate nucleus, septum, nucleus accumbens, neocortex and hippocampus were particularly enriched in 3-hydroxyanthranilic acid oxygenase immunoreactive cells. In the thalamus, regional differences were noted with regard to the intensity of staining among glial cells with high densities of 3 hydroxyanthranilic acid oxygenase cells in the anteroventral, reticular and ventromedial nuclei. In the inferior and superior colliculi, numerous 3 hydroxyanthranilic acid oxygenase-positive glial cells were found in all layers. In the hypothalamus, 3-hydroxyanthranilic acid oxygenase-immunoreactive glial cells were encountered in the zona incerta, the lateral hypothalamic area, the caudal preoptic region and in the dorsomedial nucleus. In the mesencephalon, the substantia nigra contained numerous, moderately stained cells. At caudal levels of the brain-stem, a relatively large number of cells was detected in the nucleus of the solitary tract, the pontine nucleus and in the fascial nerve nucleus, while other nuclei, such as the reticular formation and the area postrema were relatively poor in 3-hydroxyanthranilic acid oxygenase-immunoreactive cells. In addition to staining of glial cells, neuronal cell bodies containing 3 hydroxyanthranilic acid oxygenase immunoreactivity were detected in the main and in the accessory olfactory bulb, as well as in the ventromedial nucleus of the hypothalamus. Quinolinic acid phosphoribosyltransferase immunoreactivity was observed within glial cells and in association with neuronal cell bodies. Some, but not all, quinolinic acid phosphoribosyltransferase positive glial cells contained glial fibrillary acidic protein (Kohl PMID- 2974129 TI - Back pain: it should not have to be a part of the job. PMID- 2974130 TI - [Current concepts related to the etiology and pathogenesis of clefts]. PMID- 2974131 TI - Depressed mood in chronic low back pain: relationship with stressful life events. AB - This study investigated the relationship between stressful life events and depressed mood in chronic low back pain (CLBP), using both self-report and observer-rated assessments of life happenings and depression. We hypothesized that CLBP patients with depressed mood (N = 15) would report significantly more untoward life events and ongoing life difficulties compared to CLBP patients without depressed mood (N = 17) and controls (N = 19). This prediction was confirmed. Subjects also were rated as being either in a high stress or low stress condition. Patients with depressed mood were more likely to be in the high stress condition than were either non-depressed patients or volunteers. Furthermore, the increased stress reported by the distressed group appeared to be a direct consequence of back pain-related life events, rather than from other life problems. We conclude that previously reported associations between life events and CLBP are a function of the relationship between stressful life events and depressive symptoms, which are prevalent in CLBP. PMID- 2974132 TI - [The role of dehydroepiandrosterone and pregnenolone in the expression of stress behavior towards lactating females in mice]. AB - Triads of castrated male mice were chronically administered with either oil vehicle or 280 nmol of dehydroepiandrosterone (D) or pregnenolone (P). They were tested for their attack on a lactating intruder female introduced in their home cage 2, 24 or 47 hr after their last injection. D significantly reduced male aggressive behavior for at least 24 hr. Other groups of castrated males were daily treated with vehicle or 280 nmol of D, dehydroepiandrosterone sulfate (DS) or androstenediol (ADIOL), a D metabolite with clear-cut oestrogenic properties. D, but neither DS nor ADIOL, significantly reduced their aggressive responses to lactating intruders. Finally, neural levels of D, DS and testosterone (T) were measured in intact and castrated males injected with either vehicle or D. Measurable amounts of D and DS were detected, with DS being the predominant chemical form. D and DS concentrations were unchanged by castration but neural D was increased more than twenty fold in castrates treated with D, whereas DS was unchanged. The concentration of T in the brain of the intact (sham-operated) mouse approached 3 ng/g but fell close to the detection limit after gonadectomy. D treatment caused a slight but significant increase in brain T concentration in castrated mice, although T remained far below the level measured in intact males. PMID- 2974133 TI - [Function of the kidneys and ATPase activity in them in nephrotic acute renal insufficiency]. PMID- 2974134 TI - Children with chronic illness. The prevention of dysfunction. AB - Children with long-term illnesses are at risk of developing problems in psychological adjustment and in functioning in activities of daily life. Their families face increased risks of marital and economic dysfunction, and siblings too face special tasks living with a chronically ill child. A variety of interventions can help children and families to cope effectively with the tasks of chronic illness. Pediatricians should be alert to effects on the family. Children respond to family stress in very predictable ways. Inasmuch as the stress of chronic illness may affect the marital relationship, there is a likelihood of concurrent behavioral and school problems. Relatively sudden changes in behavior may signal family issues that require professional attention. Drotar et al. maintain that professionals should serve as guides or advocates for children with chronic illness and their families. The relationship that develops between families and professionals is based on trust. They believe that "trust appears to evolve from the following principles: (1) continuity of relationship, (2) active participation by professional caregivers, (3) mutual participation of child and family, (4) advocacy, (5) a focus on coping and competence, (6) a developmental perspective, and (7) a family-centered focus." Cadman et al. identified a similar set of elements that characterizes an efficacious preventive intervention approach. In addition, they propose specific programmatic efforts that are associated with less morbidity. These include ongoing education and counseling for the child, family, and community regarding chronic illness and its management, use of stress management techniques to promote mastery and reduce the impact of stressful life events, and facilitation of social support mechanisms for families with chronically ill children. We have added consideration of the child's performance in school. PMID- 2974135 TI - [Hydropericardium in hypothyreosis--a possible cause of cardiomegaly]. PMID- 2974136 TI - A case of metachronous aneurysm of the infrarenal abdominal aorta. AB - The case history of an 84 year old woman presenting with a metachronous aneurysm of the infrarenal abdominal aorta is presented. At operation this second aneurysm was replaced by a further straight tube Dacron inlay graft. This report raises the important question as to whether the whole of the infrarenal aorta should be replaced when resecting an infrarenal abdominal aortic aneurysm, rather than the more conventional approach of suturing the graft to the neck of the aneurysm above and the aortic bifurcation below. PMID- 2974137 TI - [Should all primary hyperparathyroidism be treated surgically?]. PMID- 2974138 TI - [Cardiac imaging in 1988]. PMID- 2974139 TI - [Role of allogeneic transplantation of bone marrow in juvenile chronic myelomonocytic leukemia]. AB - Two boys with clinical and haematological evidence of juvenile chronic myelomonocytic leukaemia had no chromosomal anomaly. In addition, one presented with an unbalanced Epstein-Barr virus serology, and the other with xantholeukaemia. Allogenic bone marrow transplantation was performed in the first boy after an 18-month period during which treatment with 6-mercaptopurine, intensive chemotherapy and splenectomy had failed. Conditioning included cyclophosphamide, high-dose cytarabine and whole-body irradiation. There was no complication, and 16 months after transplantation the patient was in complete remission. The second boy received a bone marrow transplant on the 6th month of the disease, after failure of 6-mercaptopurine. Conditioning included etoposide, busulfan and cyclophosphamide. On the 35th post-transplantation day the child had severe pancytopenia and his spleen remained enlarged. A second transplantation was performed after treatment with melphalan and whole-body irradiation. Twelve months later, the patient was in complete remission. The indications and modalities of allogenic bone marrow transplantation in juvenile chronic myelomonocytic leukaemia and the value of pre-transplantation splenectomy are discussed. PMID- 2974140 TI - [Pregnancy: a way of recovering from prolactin-secreting adenoma? 18 cases]. AB - Fifteen patients with prolactin-secreting pituitary adenoma completed a total of 18 pregnancies. Changes in blood prolactin levels and computerized tomographic images of the pituitary gland were studied. No increase in size of the adenoma was observed, and in 2 cases the tumour was found to be necrotic in the post partum period. In all patients the prolactinaemia significantly decreased immediately after delivery. Thus, by inducing partial necrosis of the adenoma pregnancy may result in post-partum reduction of blood prolactin levels. PMID- 2974141 TI - [Does Swan-ganz catheter help in weaning from controlled ventilation after mitral valve replacement?]. AB - The difference between oxygen consumption during spontaneous and controlled ventilation represents the oxygen cost of breathing. Swan-Ganz catheters enable oxygen consumption to be calculated as the product of cardiac index by arteriovenous oxygen concentration. This method was used in 25 patients on the day after anaesthesia and mitral valve replacement to predict the success of weaning from mechanical ventilation. Four patients could not be weaned: one for pulmonary reasons, one for neurological reasons and two because of their cardiac status. Whether weaning was successful or not, oxygen consumption did not change significantly during spontaneous respiration; concomitantly lactacidaemia remained stable. Two patients had an oxygen consumption below the physiological threshold (115 ml.min-1.m-2): one could be weaned, but not the other. Haemodynamic parameters did not change significantly during weaning. Thus, measurement of oxygen consumption through a Swan-Ganz catheter does not predict the success of weaning after mitral valve replacement, but the catheter can diagnose a low cardiac output needing treatment before weaning. PMID- 2974142 TI - [Surgery of carotid stenosis combined with contralateral obstruction. Technic of carotid bypass]. AB - Clamping of a carotid artery carries a considerable risk when the contralateral vessel is occluded. In a continuous series of 14 patients with carotid stenosis associated with contralateral occlusion, the possibility of substitute blood flow during clamping of the stentic vessel was evaluated by intra-operative measurement of carotid back pressure. In the 7 patients whose back pressure was higher than 40 mmHg, endarterectomy of the carotid bifurcation was performed in 6 cases and reimplantation for stenosing loop in 1 case. In the 7 patients whose carotid back pressure was lower than 40 mmHg, a carotid-carotid bypass was performed, the first anastomosis always being on the common carotid artery. No post-operative neurological accident was recorded. This technique seems to be preferable to a shunt, which is not always feasible, notably in cases with high bifurcation. PMID- 2974143 TI - [Absence of class I HLA antigen in acute myeloblastic leukemia]. PMID- 2974144 TI - [Spontaneous bacterial peritonitis caused by Acinetobacter calcoaceticus. A rare nosocomial infection in the cirrhotic patient]. PMID- 2974145 TI - [Actinobacillus actinomycetemcomitans whitlow]. PMID- 2974146 TI - [Tumoral tracheobronchial desobstruction by cryotherapy using a flexible catheter]. PMID- 2974147 TI - [Renal arterial thrombosis complicating nephrotic syndrome]. PMID- 2974148 TI - [Acute renal insufficiency in Mycoplasma pneumoniae pneumonia]. PMID- 2974149 TI - [Morvan's chorea caused by treatment with gold salts]. PMID- 2974151 TI - [Conditions and complications of endotracheal intubation performed outside of the hospital. A preliminary prospective study]. PMID- 2974150 TI - [Does selective inhibition of prostaglandins exist? Apropos of hyponatremia with sulindac]. PMID- 2974152 TI - [Hepatic biopsy ultrasonically guided with an "Ingenor" thick needle in the histologic diagnosis of focal hepatic lesions]. PMID- 2974153 TI - [Tuberculosis of bone and lymph nodes. A case]. PMID- 2974154 TI - [Correction of bleeding time after administration of desmopressin in a woman treated with ticlopidine]. PMID- 2974155 TI - [Relation of the infection incidence among children to the epidemiologic hazard of foci of tuberculosis infection]. PMID- 2974156 TI - [The nature of bacterial shedding and its effect on treatment outcome in newly detected patients with destructive pulmonary tuberculosis]. PMID- 2974157 TI - Polarized apical distribution of glycosyl-phosphatidylinositol-anchored proteins in a renal epithelial cell line. AB - Polarized epithelial cell monolayers contain two distinct plasma membrane domains as delineated by the presence of tight junctions--i.e., an apical surface that faces the external environment and a basolateral surface that functions both in cell-cell contact and cell-substrate attachment. Central to the understanding of epithelial cell polarity is the question of how such cell-surface specializations are generated. A different class of membrane glycoproteins has recently emerged that may yield new insight into the mechanism underlying the biogenesis of this polarity. Members of this class contain a large extracellular protein domain linked to the membrane via glycosyl-phosphatidylinositol. Using a polarized renal epithelial cell line (Madin-Darby canine kidney), we identified endogenous glycosyl-phosphatidylinositol-anchored proteins through release by a phosphatidylinositol-specific phospholipase C. Six glycosyl-phosphatidylinositol anchored proteins of 110, 85, 70, 55, 38, and 35 kDa were identified and appeared to be restricted to the apical surface. Our data are consistent with the notion that the glycosyl-phosphatidylinositol membrane anchor may contain the necessary information for "targeting" to the apical surface. PMID- 2974158 TI - Rapid detection of human chromosome 21 aberrations by in situ hybridization. AB - Plasmid clones containing up to 94 kilobases of single-copy DNA from band q22.3 of chromosome 21 and a complete pool of insert DNA from a chromosome 21 recombinant library have been used to rapidly detect numerical and structural aberrations of chromosome 21 by in situ hybridization in both metaphase and interphase cells. A trisomic karyotype, diagnostic of Down syndrome, is readily detected in nonmitotic cells because the majority of their nuclei exhibit three discrete foci of hybridization, in contrast to normal diploid cells, which show two foci. Chromosomal translocations involving chromosome 21 sequences were also detected with these probes, and the intranuclear location of 21q22.3 DNA sequences in "normal" human brain neurons was established with the plasmid DNA probe set. These results suggest that chromosome 21-specific probes may have utility in clinical diagnostics, especially by facilitating the direct analysis of interphase cells. PMID- 2974159 TI - Some mismatch repair activities in Escherichia coli. AB - Heterozygous bacteriophage lambda DNA molecules, whose replication requires mismatch correction of a mutant nucleotide in the transcribed strand, provide an assay for localized mismatch repair in Escherichia coli. We describe two systems: one removes the A in C.A or G.A mismatches and the other removes one or the other C in a C.C mismatch. Mutations disabling the first system result in a mutator phenotype that may be identical to mutY. PMID- 2974160 TI - Activation of human T lymphocytes: differential effects of CD3- and CD8-mediated signals. AB - T cells are activated physiologically by triggering the T-cell receptor-CD3 complex. There is evidence that invariant accessory molecules on the T-cell membrane (CD8 and CD4) are involved in the major histocompatibility complex restricted recognition process. Moreover, binding and crosslinking of these accessory molecules to the T-cell receptor-CD3 complex exerts a positive synergistic signal, as has been shown by stimulation with crosslinked antibodies. Here we demonstrate that stimulation mediated by immobilized anti-CD3/CD8 antibodies differs from stimulation mediated solely by anti-CD3. Whereas interleukin 2 receptor expression and interferon gamma production are seen to a similar extent in both cases, a second signal provided by the additional involvement of CD8 seems to be essential for interleukin 2 production and full interleukin 2 responsiveness in CD8+ T cells. This second signal is much more sensitive to inhibition by 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine, an inhibitor of protein kinase C and cGMP/cAMP-dependent kinases. Our results also show that substantial modulation of the T-cell receptor complex and most likely CD3 phosphorylation are not essential for initiating the activation of resting T cells. Instead, we found a 22- to 24-kDa phosphoprotein whose strong phosphorylation correlated reliably with T-cell activation. PMID- 2974161 TI - Human T-cell antigen receptor (TCR) delta-chain locus and elements responsible for its deletion are within the TCR alpha-chain locus. AB - Individual T cells express the CD3 molecule in association with alternative gamma delta or alpha beta heterodimeric T-cell receptors (TCRs). T-cell precursors and occasional gamma delta-expressing T cells in humans possess an unexpected 2.0 kilobase (kb) mRNA in which a tandemly repeated motif, TEA (T early alpha), has been spliced to the constant (C alpha) region. Long-range pulsed-field gel mapping as well as molecular cloning showed that TEA is located immediately 5' to the most upstream joining (J alpha) segment of the TCR alpha-chain locus. The TCR delta-chain locus is immediately 5' to TEA, and diversity (D delta) gene segments, J delta, C delta, and TEA are linked within 35 kb. The human TCR delta locus conserves a 12/23-base-pair (bp) spacer paradigm in which J delta possesses a 12-bp and V delta a 23-bp spacer, while the D delta segments have a 12 bp-D delta-23 bp spacer motif. Considerable TCR delta diversity can be generated despite the predominant use of one V delta and one J delta segment. Two D delta segments, D delta 1 and D delta 2, are 9 and 13 bp long, are frequently recombined as D delta 1-D delta 2, and reveal exonucleolytic trimming with extensive N-segment addition. A gamma delta clonal T cell possessed an effective VDDJ delta rearrangement and an intermediate DDJ delta rearrangement, arguing that the TCR delta locus displays allelic exclusion. Specific rearranging elements that delete the delta locus, delta Rec and psi J alpha, were mapped and found to separate the delta locus from the alpha locus. The delta locus including D delta 1-D delta 2-J delta 1-C delta-TEA was deleted in mature, alpha beta expressing T cells, whereas V delta 1 was frequently retained. The location of the delta locus within the alpha locus may necessitate an exclusive choice between delta or alpha expression. PMID- 2974162 TI - Molecular cloning and chromosomal localization of the human T-cell receptor zeta chain: distinction from the molecular CD3 complex. AB - The T-cell antigen receptor (TCR) is a multisubunit receptor complex specific to T cells subserving both antigen recognition and signal transduction functions. The zeta chain of the TCR is a component of all surface receptor complexes. This chain was first identified in murine T cells by virtue of the fact that it coimmunoprecipitates with the TCR complex using antibodies directed against either the clone-specific subunits or invariant CD3 subunits of the receptor. Recently, we have isolated a cDNA encoding the murine zeta. Using this as a probe, we have now isolated cDNAs encoding the human zeta. Sequence analysis of cDNAs encoding human and murine zeta reveals that it is a highly conserved protein. In addition to amino acid homology, there is remarkable interspecies conservation in the nucleotide sequence of the 5' and 3' untranslated regions of the zeta mRNA. The previously characterized invariant delta, epsilon, and gamma chains of the TCR, referred to as the CD3 complex, share significant sequence and structural homology with each other and are all located within 300 kilobases of each other on human chromosome 11 (11q23). zeta has no sequence similarity to the CD3 chains and the localization of the human zeta gene to the centromeric region of chromosome 1 underscores the fact that it is a distinct genetic component of the TCR. PMID- 2974165 TI - Subcutaneous tissue of the trunk and lower extremities. PMID- 2974164 TI - Early expression of a T-cell receptor beta-chain transgene suppresses rearrangement of the V gamma 4 gene segment. AB - beta transgenic mice have a T-cell receptor beta-chain gene that is prematurely expressed on the surface of CD4- CD8- thymocytes and paired with an uncharacterized non-T-cell receptor alpha-chain polypeptide. The rearrangement of the T-cell receptor variable region gamma chain gene segment V gamma 4, a component of the gamma-chain gene that is rearranged and expressed preferentially on thymocytes of normal adult mice, is severely repressed in beta transgenic mice. Consequently no gamma delta T-cell receptor heterodimers are detectable on the surface of adult thymocytes or splenic T cells. These results indicate that cells expressing alpha beta or gamma (V gamma 4)-delta TCRs originate from a common precursor in which the first productive rearrangement of either the beta or gamma locus determines the further differentiation pathway into either alpha beta or gamma delta T cells. The repression of V gamma 4 rearrangement by a preexisting beta-chain gene may be indicative of one of several mechanisms which ensure that gamma delta and alpha beta receptors do not as a rule appear on the surface of the same cell. PMID- 2974163 TI - Human T-cell-receptor delta chain: genomic organization, diversity, and expression in populations of cells. AB - The locus of the delta chain of the human T-cell receptor has been isolated and examined. Three D (diversity) regions and two J (joining) regions are present on the 5' side of the C (constant) region. The closest V (variable) region to the constant region is V delta 2, which in the germ line is found on the 3' side of the constant region in an inverted direction. The genomic structure of the human locus closely parallels its mouse counterpart. Several cDNA sequences and a series of rearranged genomic sequences are compared which demonstrate an enormous potential diversity in the junctional region, between the variable region and the joining region. We find the predominant utilization of the PEER variable region in thymic polyclonal gamma delta cell lines and in some peripheral blood gamma delta cell lines. Thus, the delta chain may have relatively limited variable region diversity but a large junctional-region diversity. The implications of this observation are discussed. PMID- 2974166 TI - Abdominoplasty: a new concept and classification for treatment. AB - From a study of the deformities of each layer of the abdominal wall, we have categorized five types of abdominoplasties. For each type, we used a different surgical technique, aiming to sculpture the abdomen and treat each layer according to the deformity present in each patient. Two hundred thirty-eight patients were treated with this method, and the results were judged good to excellent by the surgeons and patients because of the more natural appearance of the final results. PMID- 2974167 TI - Renovascular hypertension: the small kidney updated. AB - We report the results of treatment in 57 patients with renovascular hypertension associated with one poorly perfused, small kidney with less than 25 per cent of total 131I-hippurate uptake shown by renography. Arteriography in 29 patients demonstrated occlusion of the artery of the small kidney, and in 28 there was stenosis. In addition, 25 patients had stenosis of the artery supplying the larger contralateral kidney. Stenosis of the arteries of the contralateral kidneys was dilated by percutaneous transluminal arterioplasty in all but one of the 25 patients with bilateral disease of the artery, stenosis of the small kidney could be dilated successfully by percutaneous transluminal arterioplasty in 22 of the 28 patients, and cure or improvement of blood pressure was achieved in 12 of them. Percutaneous transluminal arterioplasty of occluded arteries was generally unsuccessful. In 17 patients with unilateral disease not manageable by percutaneous transluminal arterioplasty, nephrectomy of the small kidney improved blood pressure control without significant deterioration of renal function. Renal function improved in 10 patients with bilateral lesions treated by nephrectomy of the small kidney in combination with contralateral percutaneous transluminal arterioplasty. Histological examination of excised kidneys showed large infarcts or several cholesterol emboli whether percutaneous transluminal arterioplasty had been attempted or not. After observation periods ranging from two to 79 months, 48 patients were normotensive (21 without and 27 with medication) and nine patients were still hypertensive even with medication. This study showed that by using percutaneous transluminal arterioplasty initially if possible, supplemented with nephrectomy and/or medication, normotension without loss of renal function or immediate serious complications could be obtained in the majority of these severely hypertensive patients. PMID- 2974168 TI - [Antibody induced lesions]. PMID- 2974169 TI - Complement and disease. PMID- 2974170 TI - [Immunoregulation during aging]. PMID- 2974171 TI - Genetic conditioning of the in vitro human lymphocyte sensitivity to cyclosporin A. PMID- 2974172 TI - [Hydrotherapy in a warm-water pool]. PMID- 2974173 TI - Optimal placebo response rates for comparing two binomial proportions. AB - This paper considers the problem of comparing proportions in clinical trials where two parallel groups are studied. The question of what will happen to the response rate of an active therapy if some external factor affects the placebo rate is asked and two different, but similar, answers are proposed. Assuming either a logistic or probit model has substantial influence on the sample size required to detect differences and optimal comparisons are suggested. The idea is illustrated by a clinical trial of treatment for peptic ulcer and suggestions for other possible applications are presented. PMID- 2974174 TI - Laser Doppler flowmetry in evaluation of lower limb resting skin circulation. A study in healthy controls and atherosclerotic patients. AB - Laser Doppler flowmetry (LDF) was used to evaluate lower limb resting skin perfusion in sixty subjects divided into four groups: healthy young and elderly controls, and patients with intermittent claudication or critical ischaemia. Measurements were performed in pulp skin containing microvascular AV anastomoses and in the skin of leg and thigh where these shunts are absent. In toe pulp controls and claudicators had higher perfusion values than in leg and thigh skin (p less than 0.01), indicating that the LDF method evaluates flow both in nutritional capillaries, AV anastomoses and in dermal vascular plexa. Elderly controls had higher flux values in the pulp than claudicators (p less than 0.01), and claudicators had higher values than patients with critical ischaemia (p less than 0.01), showing that LDF could differentiate between the clinical groups. Study of reproducibility confirmed that values were reproducible on a given population. Day to day variation was considerable in individual subjects, probably because of changes in sympathetic vascular tone and because of different vascular architecture in the measuring volumes which are only some few mm3. The fact that LDF measures total skin blood flow explains why several papers have found a poor correlation between LDF and methods which mainly evaluate nutritional blood flow. The method is non-invasive, continuous and easy to perform. Laser Doppler flowmetry may have several clinical applications, like evaluating progress of atherosclerotic disease or therapeutic effects of drugs or operations. To increase the reproducibility of resting skin flux measurements local heating of the skin is recommended and the measurements should be performed with an integrating probe, which averages the readings obtained at several positions simultaneously. PMID- 2974175 TI - Cytostatic drug therapy in disseminated colorectal cancer. AB - 5-FU is the best available single drug in advanced colorectal disease. After systemic administration approximately 15% of the patients achieve a short-lived objective remission. Survival is not prolonged by therapy. Several chemotherapy combinations have in preliminary phase II studies given higher response rates (30 50%), but when evaluated in controlled studies, no advantage over single-drug 5 FU has been verified. At present, two combinations, MOF-S and sequential MTX/5 FU/Leucovorin seem promising, but more experience is needed before general acceptance. In cases of liver dissemination, regional hepatic infusion therapy causes objective tumour regression more often than systemic therapy. However, median survival is most likely the same for both treatment modalities, and since hepatic artery infusion is the most expensive therapeutic modality, routine use of hepatic artery infusion is discouraged. Outside clinical trials, systemic or regional chemotherapy is hardly indicated in advanced colorectal disease apart from certain selected symptomatic patients. PMID- 2974176 TI - The use of backscattered electron imaging, X-ray microanalysis and X-ray microscopy in demonstrating physiological cell death. AB - The cytochemical localization of enzymatic activity by means of backscattered electron imaging (BEI) is reviewed and the application of BEI to changes in acid phosphatase and ATPase distribution during physiological (programmed) cell death in Heliothis midgut is explored. Programmed cell death entails the release of nascent free acid phosphatase as extracisternal hydrolase. This shift can readily be detected by means of the atomic number contrast imparted by BEI of the lead phosphatase reaction product, thus enabling the distribution of dying cells to be mapped. BEI is particularly useful in this context as it allows the examination of bulk specimens at low magnification. Death of cells is also accompanied by a collapse in ATPase activity which shows up as cytochemically negative areas in the X-ray microscope and by means of BEI. Acid phosphatase in normal cells is localized in the apical microvilli and lysosomes. Senescent or dying cells, however, clearly show a basally situated free hydrolase which migrates throughout the cell. Parallel TEM results confirm that this enzyme is ribosomal and extracisternal rather than lysosomal in origin. ATPase activity is largely limited to the apical microvilli, although there is some activity associated with the basal plasma membranes. The apical ATPase, however is partially resistant to ouabain. Young and mature cells are positive although in the latter case some microvilli may be lost as the cells acquire a negative cap or dome. Inhibition by bromotetramizole indicates that apical activity is not to any significant extent contributed to by alkaline phosphatase. Degenerate or dead cells are negative and can be seen as a mozaic of "black patches" among normal cells when imaged by means of BEI or X-ray microscopy. PMID- 2974177 TI - Scanning electron microscopic autoradiography of lung. AB - Scanning electron microscopic (SEM) autoradiography of the lung is being used to determine the distribution of inhaled, alpha particle-emitting, plutonium dioxide particles. SEM autoradiography provides high visual impact views of alpha activity. Particles irradiating the bronchiolar epithelium were detected both on the bronchiolar surface and in peribronchiolar alveoli. The technique is being used to obtain quantitative data on the clearance rates of plutonium particles from bronchi and bronchioles. PMID- 2974178 TI - [DNA analysis in the assessment of leukemia]. AB - In 5 cases of leukaemia and in two other cases where a diagnosis of leukaemia was considered, no definitive classification was made by morphology, cytochemistry, patterns of surface markers and/or cytogenetics. Therefore, genotyping was performed by DNA analysis of leukaemic cells. Somatic rearrangements in leukaemic cell DNA of immunoglobulin and T cell receptor genes, and of the breakpoint cluster region on chromosome 22 involved in the Philadelphia chromosome translocation, proved to be useful DNA markers to confirm or refute particular types of leukaemia. PMID- 2974179 TI - Cyclic AMP-responsive DNA-binding protein: structure based on a cloned placental cDNA. AB - Cyclic AMP (cAMP) is an intracellular second messenger that activates transcription of many cellular genes. A palindromic consensus DNA sequence, TGACGTCA, functions as a cAMP-responsive transcriptional enhancer (CRE). The CRE binds a cellular protein of 38 kD in placental JEG-3 cells. A placental lambda gt11 library was screened for expression of specific CRE-binding proteins with the CRE sequence as a radioactive probe. A cDNA encoding a protein of 326 amino acids with the binding properties of a specific CRE-binding protein (CREB) was isolated. The protein contains a COOH-terminal basic region adjacent to a sequence similar to the "leucine zipper" sequence believed to be involved in DNA binding and in protein-protein contacts in several other DNA-associated transcriptional proteins including the products of the c-myc, c-fos, and c-jun oncogenes and GCN4. The CREB protein also contains an NH2-terminal acidic region proposed to be a potential transcriptional activation domain. The putative DNA binding domain of CREB is structurally similar to the corresponding domains in the phorbol ester-responsive c-jun protein and the yeast transcription factor GCN4. PMID- 2974180 TI - [Involvement of the arcuate nucleus of the hypothalamus in the descending pathway from the nucleus accumbens to the periaqueductal gray subserving an antinociceptive effect]. PMID- 2974182 TI - [Autodermoplasty of recurrent large and gigantic hernias]. PMID- 2974181 TI - Differential diagnosis of severe back pain using MRI. AB - Back pain, despite its prevalence, often presents a diagnostic dilemma. Infection, degeneration, and neoplasm comprise major etiologic categories of severe nonspecific back pain. Diagnostic evaluation includes plain roentgenograms, computerized tomography, and radionuclide studies, all of which are often equivocal or misleading. We retrospectively analyzed 21 presentations of severe back pain of various causes evaluated by magnetic resonance imaging (MRI) in addition to conventional diagnostic imaging modes. A characteristic MRI pattern of both the lesions's distribution and its signal intensity was observed that delineated each etiologic category. MRI was found to be particularly suited for use in the differential diagnosis of nonspecific back pain. PMID- 2974183 TI - [Clinico-epidemiologic characteristics of salmonellosis]. PMID- 2974184 TI - [Percutaneous transluminal angioplasty of cerebrovascular stenoses-- indications, technic, long-term results]. PMID- 2974185 TI - [Duplex sonographic follow-up after carotid thrombendarterectomy and patch plasty]. PMID- 2974186 TI - [Modification of the autonomic vascular reaction and peripheral circulation in the handgrip and orthostasis test by beta-blockage in patients with and without peripheral arterial occlusive disease]. PMID- 2974187 TI - [2 years follow-up following laser angioplasty of peripheral arterial obstructions]. PMID- 2974188 TI - [Percutaneous transluminal angioplasty of subclavian stenoses]. PMID- 2974189 TI - [Laser angioplasty--experiences after 15 months]. PMID- 2974190 TI - [Laser angioplasty: sapphire tip versus hot tip]. PMID- 2974191 TI - [Laser percutaneous transluminal angioplasty--experiences up to now, problems and early results]. PMID- 2974192 TI - [Fibrinolytic therapy of thromboembolic vascular occlusions: experiences based on 87 patients]. PMID- 2974193 TI - A rapid, simple method for monitoring fibrinolysis in vitro. PMID- 2974194 TI - Quantification of the sulfates of 16 alpha-hydroxy androgens that are possible precursors of estriol-3-sulfate in human breast cyst fluid. AB - The concentration of 16 alpha-hydroxydehydroepiandrosterone-3-sulfate (16 alpha OHDHAS) was determined in 29 samples of human breast cyst fluid (BCF) and in 15 of these, androst-5-ene-3 beta,16 alpha,17 beta-triol-3-sulfate (A-TriolS) was also assayed. The median value of both was about 100 ng/mL and the ranges were from 1.4 to about 1800 ng/mL. There was a significant association in the values for the two sulfates (p less than 0.05). These concentrations are consistent with a role for 16 alpha-hydroxy androgens as possible precursors for estriol-3 sulfate. The latter is highly elevated relative to other body fluids in BCF. The androgens also correlated directly with the concentrations of K+, an indicator of apocrine proliferation of breast cysts. PMID- 2974196 TI - Kidney graft survival and immunologic changes in patients conditioned with donor specific transfusions prior to transplantation. PMID- 2974195 TI - [Paraclinical examinations]. PMID- 2974198 TI - Induction of suppressor T cells by cytokines. PMID- 2974197 TI - Molecular characterization of suppressor T cells. PMID- 2974199 TI - Complement activation during blood donation. PMID- 2974200 TI - Differential immunoregulatory effect of allosensitized suppressor T cells on CD4- and CD8-positive T lymphocytes. PMID- 2974201 TI - Suppressor cells and mediators as immunoregulators. PMID- 2974203 TI - The relationship between hypertension and left ventricular hypertrophy in renal transplant recipients. PMID- 2974202 TI - A comparison of single and multiple blood transfusions in the mixed lymphocyte response of rats. PMID- 2974204 TI - An overview of the use of the monoclonal antibody OKT3 in renal transplantation. AB - The murine monoclonal anti-T cell antibody, OKT3, has been used during the past 6 years in various clinical transplantation studies. Undoubtedly it has proved to be the most effective and specific immunosuppressive drug widely available for use in organ transplantation. However, because of problems of first-dose symptoms, recurrent rejection, and antibody production, currently used protocols with OKT3 might not be the final answer in immunosuppressive therapy. OKT3 is an important prototype of future monoclonal antibodies and can be considered the gold standard against which newer drugs must be compared. PMID- 2974205 TI - Dissociated expression of receptors for interleukin 2 and transferrin in the presence of preformed suppressor T cells and interleukin 2. AB - In vitro activated suppressor cells were tested for their ability to inhibit TFR expression. Interleukin 2 receptors and TFR are similarly suppressed on both CD4+ and CD8+ T cells. Addition of exogenous rIL2 permits appearance of IL2R but not TFR on CD4+ and CD8+ T cells. In the presence of rIL2, however, the percentage of CD4+ T cells is significantly diminished, while Leu11-positive natural killer cells escape the regulatory effect of Ts. PMID- 2974206 TI - Catalase increases lymphocyte proliferation in mixed lymphocyte culture. PMID- 2974207 TI - [Use of systematic echocardiography during a rheumatic flare-up (apropos of 67 cases)]. PMID- 2974208 TI - [The place of laparoscopy in the diagnosis of cancer of the gallbladder]. PMID- 2974209 TI - [The role of tomodensitometry in the study of superficial metastases: description of clinical cases]. PMID- 2974210 TI - [Fetal stress response during pregnancy and delivery. Adrenaline, noradrenaline and human beta-endorphin]. PMID- 2974211 TI - Automated calculation of stenosis diameters from the width of the velocity jet with the use of a multi-gate pulsed Doppler system. AB - The aim of this study was to evaluate an algorithm for automated estimation of the width of a jet stream originating from a stenosis. The evaluation was performed in a pulsatile flow model. The width of the jetstream was assessed by measuring the diameter of the region with relatively high velocities (the jet) in the velocity profiles, as recorded with a multi-gate pulsed Doppler system. Measurements were performed at 3, 6, and 9 mm downstream of three different stenoses (stenosis diameter: 3, 5, or 8 mm) at different Reynolds numbers (200 1600) based on time averaged flow velocity for a tube of diameter 15 mm. The developed algorithm was used successfully for automated detection and quantification of jet flow diameters downstream to a stenosis. The algorithm can be used for calculating the stenosis diameter notwithstanding a theoretically predictable overestimation of about 1 mm, depending on the Reynolds number and the distance from the stenosis. PMID- 2974212 TI - Maximum likelihood frequency tracking of the audio pulsed Doppler ultrasound signal using a Kalman filter. AB - Accurate estimation of velocity from the audio Doppler signal is important in the study of disease. The paper addresses this problem by considering the application of maximum likelihood as the basis for frequency tracking using a Kalman filter. The effectiveness of the algorithm in velocity estimation on both test signals and clinical data is discussed. This is particularly important in tracking of the oscillations which occur in the deceleration phase of the cardiac cycle. PMID- 2974213 TI - Netobimin in drinking water for treatment of bovine parasitic bronchitis: a field experiment. AB - Lungworm-infected seeder calves were used on four 1.41 ha paddocks to ensure that groups of 11 calves would be exposed to a heavy challenge with Dictyocaulus viviparus. By the 39th day after turnout there was a serious episode of respiratory disease and a diagnosis of parasitic bronchitis was confirmed by post mortem and faecal examination. One group of trial calves was treated with netobimin administered in the drinking water at 2.8 mg/kg/day for seven consecutive days; another group received the same treatment supplemented with flunixin meglumine at 2.2 mg/kg/day for three days; a third group was given a single oral dose of 7.5 mg netobimin/kg; only emergency treatments were given to calves in the control group. The clinical response to the drinking water treatments was highly satisfactory and better than the response to the single oral treatment. PMID- 2974214 TI - Immunogenetic analysis of Trichinella spiralis infections in swine. AB - The immune responses of outbred swine, inoculated with several different low doses of Trichinella spiralis muscle larvae (ML), was followed over 5-6 weeks of primary infection, in order to determine an inoculation dose which could be used to identify genetic controls on the response to this helminth parasite. Reproducible infections were established when swine were inoculated with 100-300 ML. Humoral antibody responses to different larval stages were evident at 4 weeks using enzyme-linked immunosorbent assay (ELISA) of antibody-binding to excretory secretory (ES) products of ML, and flow cytometric (FCM) analysis of antibody binding to newborn larvae. T-cell blastogenesis to T. spiralis ML antigens was predominantly in the CD4+, class II restricted, T-cell subset. Having established an appropriate inoculation dose, swine leukocyte antigen (SLA) inbred miniature swine were then inoculated with this low dose of T. spiralis ML, to determine whether major histocompatibility complex (MHC) genes regulate swine immune responses to T. spiralis, as has been found in rodent models. Preliminary evidence indicated that swine of the SLA c/c haplotype may exhibit a lower burden of T. spiralis larvae in the tongue and diaphragm. This lower muscle burden correlated with the earlier development of a humoral antibody response in these genetically-defined swine. PMID- 2974215 TI - [X-ray endovascular dilatation of the brachiocephalic arteries]. PMID- 2974216 TI - [Percutaneous transluminal dilatation of stenoses of the supraaortic blood vessels]. PMID- 2974217 TI - Cutaneous T cell lymphoma: immunocytochemical study on activation/proliferation and differentiation associated antigens in lymph nodes, skin, and peripheral blood. AB - The expression of activation/proliferation antigens (CD 25, CD 30, Ki 67, transferrin receptor) in lymph nodes and skin were compared in nine patients with mycosis fungoides (MF) with patients with erythroderma not related to MF, and patients with reactive lymphofollicular hyperplasias (a total of 14 patients). A panel of differentiation antigens was analyzed in addition. Reactivities were revealed by the APAAP technique. Activation/proliferation antigen scores in lymph nodes were related to the clinical stages of MF in most instances (low scores in cases of MF stage I/II, high scores in 3/4 cases of MF stage III/IV). They differed markedly in cases of non-MF-erythrodermia with the exception of one patient, and in all cases of reactive lymphofollicular hyperplasia. Expression of activation/proliferation antigens in lymph nodes were different in most cases from those in skin and peripheral blood. For diagnostic use, the activation/proliferation antigen scores were superior to the cell differentiation antigen profiles. Among cellular differentiation antigens, only the extent of CD1+ cells provided some diagnostic information, since the number of these cells were markedly increased in all cases of dermatopathic lymphadenitis with/without MF when compared with reactive lymphofollicular hyperplasia. In the diagnosis of MF, immunohistochemistry of activation/proliferation or differentiation antigens cannot replace routine paraffin histology, but may provide supplement any data in equivocal cases. PMID- 2974218 TI - Organization of the middle RNA segment of snowshoe hare Bunyavirus. AB - The genetic organization of the M RNA segment of snowshoe hare (SSH) virus, a member of the Bunyavirus genus of the family Bunyaviridae, has been determined. The middle (M) RNA segment has a single open reading frame (ORF) of 1441 amino acids. We have used amino- and carboxy-terminus sequencing and synthetic peptides to map proteins within the ORF. The order of the proteins translated from the single large open reading frame is G2, NSm, G1. The G2 protein extends from amino acids 14 to 299. The molecule is 286 residues long, with a computed nonglycosylated molecular weight of 31,973 Da. It is preceded by a cleaved 13 amino acid signal sequence. G2 includes a long highly hydrophobic sequence and contains three potential N-linked glycosylation sites. The G1 protein occupies the C-terminal end of the open reading frame from amino acids 474 to 1441 (968 amino acid residues) and has a computed nonglycosylated, molecular weight of 108,981 kDa. It has two potential N-linked glycosylation sites, and a potential transmembrane region followed by a potential cytoplasmic domain at the C-terminal end. If membrane associated it has an orientation of N-terminus outer, C-terminus inner. Limited trypsin digestion removes a 33-kDa fragment from the N-terminal end, leaving a virion-associated truncated G1 molecule (amino acids 762 to 1441) with a single N-linked glycosylation site. Between the G2 and G1 molecules there are 174 amino acids, sufficient to code for 19 kDa of protein. Some antibodies raised against peptides within this region react with proteins of 11 kDa (NSm) and 10 kDa present in infected cell lysates, but the exact relationship of these proteins to the open reading frame remains to be determined. PMID- 2974219 TI - Identification of sequence changes in the cold-adapted, live attenuated influenza vaccine strain, A/Ann Arbor/6/60 (H2N2). AB - Nucleotide sequences have been obtained for RNA segments encoding the PB2, PB1, PA, NP, M1, M2, NS1, and NS2 proteins of the influenza A/Ann Arbor/6/60 (H2N2) wild-type (wt) virus and its cold-adapted (ca) derivative that has been used for preparing investigational live attenuated vaccines. Twenty-four nucleotide differences between the ca and wt viruses were detected, of which 11 were deduced to code for amino acid substitutions in the ca virus proteins. One amino acid substitution each was predicted for the PB2, M2, and NS1 proteins. Two amino acid substitutions were predicted for the NP and the PA proteins. Four substitutions were predicted for the PB1 protein. The biological significance of mutations in the PB2, PB1, PA, and M2 genes of the ca virus is suggested by currently available genetic data, a comparison with other available influenza gene sequences, and the nature of the predicted amino acid changes. In addition, the sequence data confirm the close evolutionary relationship between the genomes of influenza A (H2N2) and influenza A (H3N2) viruses. PMID- 2974220 TI - [Echocardiographic and Doppler signs of pulmonary hypertension and overloading of the right ventricle in left-sided valvular defects]. PMID- 2974221 TI - [Hypertrophy and left ventricular function in arterial hypertension in the echocardiographic picture]. PMID- 2974222 TI - [Percutaneous transluminal coronary angioplasty as an alternative to surgical reconstruction of the coronary vessels]. PMID- 2974223 TI - [Clinical, immunologic and therapeutic aspects of endangiitis obliterans]. AB - In the period of 1975-1983 twenty-three patients with thromboangiitis obliterans were examined at the Surgical Clinic of the Medical Academy of Magdeburg. The diagnosis was established clinically, angiographically histologically and immunologically. In 7 out of 12 patients histology revealed inflammatory vascular alterations including lymphocyte infiltrates throughout the entire vessel wall and also in the perivascular area. Immunohistology showed in 10 out of 15 patients segmental granular fluorescence identifiable as deposits of IgM and IgG. In 6 cases a complement formation was found. The endangiitis group revealed more frequent and increased immune complexes concentrations. Circulating immune complexes were established by phase-locked radio-immunoassay. The therapy is depended on the localization of the obliteration and can be a lumbar sympathectomy or a vascular reconstruction. PMID- 2974224 TI - [The Schloffer tumor. Morphology and clinical aspects of 3 cases]. AB - Three cases of the so-called Schloffer tumour are presented. It is a rare inflammatory pseudotumour of the abdominal wall with aggressive connective tissue proliferation which frequently infiltrates neighbouring abdominal organs. Usually this tumour occurs several years after abdominal surgery or trauma, thus posing substantial problems in the interpretation of the clinical and morphological findings. In one of our male patients for instance, a sarcoma of the abdominal wall was suspected pre- and intraoperatively. A female patient was falsely considered to have a carcinoma of the urinary bladder. The lesion belongs to the reactive tumour-like fibromatoses and has to be distinguished from genuine soft tissue tumours, especially the abdominal desmoid. PMID- 2974225 TI - [Studies on the structural alterations of the human thymidine kinase gene occurring in the process of cotransfection]. PMID- 2974226 TI - Lymphocyte subpopulations in malignant ascites of serous papillary ovarian adenocarcinoma. An immunocytochemical study. AB - The aim of this study was to investigate lymphocyte subpopulations in 17 patients with malignant ascites due to serous papillary adenocarcinoma of the ovary. Eight patients had not been treated prior to the study whereas nine patients had been treated by surgery and chemotherapy. A panel of monoclonal antibodies against surface markers that correlate with the immune functions of the lymphocytes was used. The lymphocyte subpopulations were identified by the immunoperoxidase adhesive slide assay, and the results in treated and untreated patients were compared. Both groups of patients showed lymphocytosis (41 +/- 25% and 33 +/- 14% of the total cells, respectively). The untreated patients had a significantly higher proportion of B cells (14 +/- 4% of lymphocytes) than did treated patients (7 +/- 2%). No differences were found between both groups regarding the helper inducer/suppressor-cytotoxic T lymphocyte ratio. The proportion of lymphocytes expressing interleukin-2-receptors was higher in treated patients (6 +/- 2%) than in untreated patients (1.2 +/- 1%). Both groups showed a high percentage of natural killer/cytotoxic cells (17 +/- 7% and 18 +/- 5%, respectively). In the only chylous effusion in this study, there was an increase in helper-inducer and activated T lymphocytes. Future studies are required to document whether surface marker analysis of lymphocytes in malignant effusions may be useful for assessment of the prognosis and the results of treatment. PMID- 2974228 TI - Calcification of the basal ganglia in Down's syndrome and Alzheimer's disease. AB - The prevalence and severity of calcification in the basal ganglia (BGC) has been examined histopathologically in 194 patients divided into ten diagnostic categories. The prevalence and severity of BGC was greater (for age) in Down's syndrome and in patients under 75 years of age with Alzheimer's disease. The severity, but not the prevalence, of BGC was greater in Down's syndrome than in patients of similar age with Alzheimer's disease. Both the prevalence and the severity of BGC in patients over 75 years of age with Alzheimer's disease were as expected for age alone. The increased prevalence and severity of BGC in Down's syndrome and in younger patients with Alzheimer's disease appeared not to be related to the presence of dementia or degenerative disease per se, nor was it affected by the presence of cerebral infarction. BGC may result from an age related disturbance of the structure of arteries within the globus pallidus, which is accelerated (or occurs prematurely) in Down's syndrome and in younger patients with Alzheimer's disease, but probably does not form part of that spectrum of changes that constitutes the pathological basis of Alzheimer's disease. PMID- 2974227 TI - Expression of the histocompatibility glycoprotein HLA-DR in neurological disease. AB - Reactive microglia or macrophages expressing the histocompatibility glycoprotein HLA-DR were detected in many neurological diseases including Alzheimer's, Parkinson's, Pick's and Huntington's diseases, parkinsonism-dementia of Guam, amyotrophic lateral sclerosis, Shy-Drager syndrome, multiple sclerosis and AIDS encephalopathy. Reactive astrocytes, also present in these conditions, were established as a population distinct from the HLA-DR positive microglia by double immunostaining for glial fibrillary acidic protein and HLA-DR. A distinctive pattern of HLA-DR positive cells was seen in each disease entity. Areas known to contain pathology always stained positively, and, in several cases, reactive microglia appeared in areas that would otherwise not have been suspected of being involved in the pathological process. HLA-DR staining, which outlines the surface membranes of positive cells, was so strong that lesioned areas could frequently be identified in sections with the naked eye. In adjacent sections stained with H&E or sections destained of HLA-DR and then restained with H&E, gliosis was often hard to identify except on close microscopic inspection. The results suggest that HLA-DR staining may be a valuable addition to standard neuropathological methods and might be useful in investigating diseases where pathology has not yet been identified. PMID- 2974229 TI - Ipratropium treatment of rhinorrhea in perennial nonallergic rhinitis. A Nordic multicenter study. PMID- 2974230 TI - Comparative study of the electrocardiograms of healthy fullterm and premature newborns. AB - The electrocardiograms of 421 healthy fullterm and premature newborns, recorded on the fifth day of life, were evaluated for possible systematic differences related to gestational age. All newborns were appropriate for gestational age and were divided into four groups according to birthweight. As birthweight increased, we noted: (a) a progressive rightward shifting of the QRS axis in the frontal plane; (b) an increase in the amplitude mainly of the precordial R and S waves reflecting the right ventricle and both ventricles combined, whereas a less significant increase or even a decrease was noted in the R and S waves reflecting the left ventricle; (c) a more frequent occurrence of diphasic and positive T waves in the right precordial leads; and (d) a prolongation in the duration of the P wave and the QRS complex. These findings reflect both an increase in total myocardial mass as well as a progressive right ventricular predominance, as birthweight or gestational age increases. Practical implications for electrocardiographic identification of cardiac hypertrophy in newborns of different gestational age are discussed. PMID- 2974231 TI - Fetal and neonatal cortical adrenal function in birth asphyxia. AB - Eighteen newborn infants, gestational age between 36 and 42 weeks with birth asphyxia were compared with 23 normal newborn infants to determine serum cortisol and dehydroepiandrosterone sulfate levels in cord blood and in venous blood samples collected 12-18 hours after birth. Both groups were similar in gestational age, birthweight, proportion of small for gestational age and large for gestational age infants, proportion of infants delivered by cesarean section with and without labor, and proportion of mothers with pre-eclampsia. There was no antenatal exposure to corticosteroid. The asphyxiated newborn infants had a significantly higher mean cord serum level of cortisol, and a significantly lower mean cord serum level of dehydroepiandrosterone sulfate than the control group. Mean serum cortisol and dehydroepiandrosterone sulfate levels collected 12-18 hours after birth were similar between both groups. It is suggested that elevated cord serum level of cortisol is related to birth asphyxia stress stimulating the adrenal definitive zone, and the low cord serum level of dehydroepiandrosterone sulfate is secondary to a transient hypoxemic-ischemic insult to the adrenal fetal zone. PMID- 2974232 TI - Atrial natriuretic factors (ANF) and antidiuretic agents. AB - The effects of lysine-vasopressin (VP) and pepsanurin (PU) on the diuretic saluretic action of ANF were investigated. Anaesthetized female rats under constant intravenous perfusion with isotonic glucose solution (0.6 ml/h/100 g body weight) were used. Blood pressure was recorded continuously and in the urine collected every 20 min, volume, NA and K excretion were measured. In each rat two intravenous boluses of either 2.5 ug synthetic rat atriopeptin II or an equivalent amount of a semipurified rat atrial extract were assayed. Thirty min before the ANF second bolus, an i.v. injection of lysine-vasopressin was given. Doses of 0.1, 1, 5, 10 and 50 mU were used. PU in the dose of 0.5 ml, obtained from 20 ml of human plasma was administered intraperitoneally, 40-60 min before the second bolus of ANF. The smaller doses of VP did not inhibit the urinary response to ANF, and the higher one (50 mU) produced a significant facilitation of ANF effect. Contrariwise, PU produced a considerable inhibition on water, NA and K excretion promoted by ANF. PMID- 2974233 TI - Coronary thrombolysis. Principles and practice. PMID- 2974234 TI - Immunological studies of patients with Down's syndrome. Measurements of autoantibodies and serum antibodies to dietary antigens in relation to zinc levels. AB - Recurrent diarrhoea and weight loss in many adult patients with Down's syndrome (DS), initiated a search for malabsorption based on determination of serum IgG and IgA antibody levels to dietary antigens. The results were compared with measurements of autoantibodies and serum zinc levels. DS patients had increased IgG and IgA activities to gluten proteins, casein and ovalbumin compared with an age- and sex-matched group of other mentally retarded patients in the same institution. Intestinal biopsy was performed in six of the 38 patients; one had total and one partial villous atrophy. Serum zinc was significantly lower in DS patients (median 14.7 mumol/l, range 5.5-20 mumol/l) than in the controls (median 16.4 mumol/l, range 12.7-19.5 mumol/l). DS patients with increased IgA activity to gluten weighed less and had lower concentrations of zinc in serum than DS patients with normal IgA activity. Twenty-eight per cent of the DS patients had autoantibodies to the thyroid gland. Our results suggest intestinal malfunction in DS, perhaps related to a defect of immune regulation caused by reduced levels of zinc in serum. PMID- 2974235 TI - Which T cells are relevant to resistance against Listeria monocytogenes infection? PMID- 2974236 TI - In vivo and in vitro role of gamma interferon in immune clearance of Rickettsia species. PMID- 2974238 TI - Planning playgrounds for children with disabilities. AB - Occupational therapists are well qualified for involvement in playground planning. They know how to eliminate architectural barriers, adapt equipment to maximize independence and functional levels, and use play in facilitating children's development. By participating in playground planning or adaptation, the occupational therapist can present play opportunities for children with disabilities, increase public awareness of occupational therapy, and extend the use of treatment modalities. This paper makes specific suggestions for planning and building to make the idea for a playground a realistic goal. A case example illustrates the planning of a playground in a hospital setting. PMID- 2974237 TI - Doxazosin: A distinctive approach to risk reduction of coronary heart disease in hypertensive patients. Proceedings of a symposium. Hamburg, West Germany, January 30, 1988. PMID- 2974239 TI - Did change of wording influence subjects' responses? PMID- 2974240 TI - The presence of heparan sulfate proteoglycans in the neuritic plaques and congophilic angiopathy in Alzheimer's disease. AB - Two immunocytochemical probes were used to specifically identify and localize heparan sulphate proteoglycans (HSPGs) in 17 cases of Alzheimer's disease (AD). A monoclonal (HK-102) and an affinity-purified polyclonal antibody, each recognizing specific domains on the protein core of a basement membrane-derived HSPG, localized HSPGs to the amyloid fibrils present in neuritic plaques (NPs) and congophilic angiopathy (CA) in the brains of Alzheimer's patients, with weak to no immunostaining in neurofibrillary tangles from the same tissues. HSPGs were also demonstrated in "primitive plaques," suggesting that their accumulation takes place during early stages of plaque development. Immunolocalization of HSPGs to subsets of astrocytes and neuronal cells, particularly those in close proximity to NPs and CA, suggested possible involvement of these two cell types in deposition of HS-PGs into the amyloidotic lesions. The current study not only identifies a new component (HSPGs) present in the amyloid deposits of NPs and CA but also suggests that astrocytes, neurons, or both may be involved in its deposition at these sites. PMID- 2974241 TI - Analysis of a murine B cell lymphoma, CH44, with an associated non-neoplastic T cell population. I. Proliferation of normal T lymphocytes is induced by a secreted product of the malignant B cells. AB - A non-neoplastic T cell population associated with a murine monoclonal B cell malignancy, CH44, was analyzed. Immunofluorescence on cell suspensions and immunoperoxidase staining on tissue sections using monoclonal antibodies to the antigens Thy1.2, Ly-1, L3T4, and Lyt-2 confirmed the presence of both TH (Ly 1/L3T4+, Lyt-2-) and Tc/s (Ly-1/L3T4-, Lyt-2+) T cell subpopulations. The non neoplastic T cells were present in both a 0.6 and 2.1 g CH44-bearing spleen. T cells, not normally in liver in significant numbers, were found in liver tissue when the CH44 tumor cells were present. These data implied an active proliferation of the T cell populations within tissues containing the malignant B cells. Supernatant from an in-vitro-adapted cell line of CH44 (CH44.LX) was tested for its ability to induce proliferation of normal murine splenocytes and thymocytes. As assayed by tritiated thymidine incorporation, both spleen and thymus cells proliferated in the presence of CH44.LX supernatant. Although supernatant from two of nine other B cell lines was able to stimulate the proliferation of spleen cells, only CH44.LX could induce proliferation of thymus cells. Supernatant from the seven other B cell lines and three hybridomas had no measurable effect on either splenocytes or thymocytes in this assay. It is hypothesized that the presence of a non-neoplastic proliferating T cell population associated with a neoplastic B cell lymphoma during in vivo passaging of the tumor is the result of effects derived from a secreted product of the malignant B cells. Whether the T cells have any effect on the growth of the malignant B cells is not known. PMID- 2974242 TI - Contribution of the kidney to metabolic clearance of atrial natriuretic peptide. AB - To quantify the role of the kidney in whole body metabolic clearance rate (MCR) from plasma of atrial natriuretic peptide (ANP), synthetic alpha-human ANP-(1-28) was infused at 200 ng/min to steady-state conditions in chronically instrumented one-kidney conscious dogs. Clearances were measured in dogs with a normally filtering kidney and they were also measured after the glomerular filtration rate (GFR) was reduced to close to zero by acutely inflating a cuff around the renal artery (RAC), which resulted in minimal urine production and renal blood flow reduction to 59% of the resting level. In normal dogs, MCR was 1,090 +/- 134 ml/min with renal clearance rate (RCR) contributing only 13.9%. After RAC, MCR fell to 864 +/- 151 ml/min, due in part to a fall in RCR (-41.5 +/- 12.9 ml/min), but mostly due to a fall in "rest of the body" (total renal) clearance of ANP. The reduced GFR accounted for virtually all the fall in RCR. Normal plasma ANP half-life was 59.6 +/- 7.9 s. In conclusion, MCR of ANP was very high, approaching the cardiac output, suggesting that most of ANP is cleared in one circulation through peripheral tissues. GFR contributed significantly to RCR (approximately 30%) but the contribution of the kidney to whole body MCR was small relative to rest of the body clearance of ANP. PMID- 2974243 TI - Histamine H2 receptor stimulation increases gastric emptying in monkeys. AB - We studied the effect of the specific H2 receptor agonist dimaprit on gastric emptying using a dye dilution technique in five chair-adapted rhesus monkeys to determine simultaneously gastric emptying and H+ secretion. Continuous subcutaneous injections of either saline alone or dimaprit (15, 30, 60, 120, and 240 nmol.kg-1.min-1) were given in random order on separate days. Both fractional emptying rate and H+ output were significantly increased by higher doses of dimaprit, with resulting twofold concurrent increase of intragastric H+ concentration. Because intragastric administration of H+ decreases gastric emptying and secretion, we evaluated the possibility that the stimulatory action of a histamine H2 agonist was underestimated when gastric pH was greater during control than after dimaprit. When exogenous HCl was added to the stomach to achieve H+ concentrations similar to those measured after dimaprit, both fractional emptying and H+ secretion were decreased twofold. When these "acidified control values" were used as a point of comparison, the stimulatory effect of dimaprit on both fractional emptying and gastric secretion was greater, suggesting that the stimulatory actions of dimaprit are underestimated if one does not take into account the differences in intragastric H+ concentrations. PMID- 2974244 TI - Renal phosphate transport in humoral hypercalcemia of malignancy. AB - Fisher rats bearing the H-500 Leydig cell tumor (LCT) develop humoral hypercalcemia of malignancy (HHM), which is accompanied by hypophosphatemia and hyperphosphaturia. To better define the mechanisms underlying the changes in phosphate metabolism, the activity of sodium-dependent phosphate uptake (Na+-Pi) in microvillus membrane vesicles (MMV) isolated from the renal cortex of LCT bearing rats was studied. Ten days after tumor transplantation the animals became hypercalcemic, hypophosphatemic, and hyperphosphaturic, and LCT-MMV showed a specific decrease in Na+-Pi. A kinetic analysis revealed evidence for both a high and a low-affinity system of Na+-Pi. The Vmax of both the low-affinity system and the high-affinity system were significantly reduced in LCT-MMV. These changes in Na+-Pi transport were similar to those induced by parathyroid hormone. In day 10 tumor-bearing animals, daily injections of dichloromethylene diphosphonate (2.5 mg.kg-1.day-1) prevented the onset of hypercalcemia but not the reduction in Na+-Pi in LCT-MMV. Our data suggest that in this animal model of HHM there is a specific and persistent impairment of Na+-Pi uptake at the level of the renal cortical brush-border membrane, which contributes to the derangement in phosphate metabolism. PMID- 2974246 TI - Endogenous ANP augments fractional excretion of Pi, Ca, and Na in rats with reduced renal mass. AB - Atrial natriuretic peptide (ANP) infusion increases fractional excretion of many solutes including sodium, chloride, bicarbonate, phosphate, calcium, and magnesium. Because fractional excretion of these solutes increases with advancing renal disease, and because plasma ANP levels are known to be elevated in chronic renal failure, we sought to determine whether ANP mediates increased solute excretion rates per nephron in rats following extensive renal ablation, a model of chronic renal failure. Because sodium restriction decreases plasma ANP levels in the setting of reduced renal mass, we also determined the effect of sodium restriction on sodium, phosphate, calcium, and magnesium excretion rates in rats with 5/6 nephrectomy (NX). We also assessed whether high endogenous ANP levels influence fractional sodium, phosphate, calcium, and magnesium excretion in rats with 5/6 NX, by inhibiting ANP action via infusion of a high-affinity ANP antiserum. Whole-kidney glomerular filtration rate in 5/6 NX rats averaged approximately one-third that of shams, and plasma ANP levels were significantly elevated in these rats above those of shams, but to a lesser extent in rats on low- vs. high-salt intakes. Fractional sodium, phosphate, and calcium, but not magnesium excretion rates were significantly greater in 5/6 NX rats on the higher sodium intake compared with those in 5/6 NX rats on the lower sodium intake. Moreover, in 5/6 NX rats on the higher sodium intake, ANP antiserum significantly reduced fractional sodium, phosphate, and calcium excretion, but was without effect on magnesium excretion. These data implicate endogenous ANP in promoting the adaptive increase in sodium, phosphate, calcium, but not magnesium excretion per nephron in chronic renal disease. PMID- 2974245 TI - Response of atrial natriuretic peptide to acute saline loading in essential hypertension. AB - To further investigate the mechanism(s) of the exaggerated natriuretic response of hypertensives to volume expansion (VE; 1,800 ml iv isotonic saline over 3 h), the plasma levels of immunoreactive atrial natriuretic peptide (ANP) were measured in 11 normal subjects (NT) and 12 patients with mild essential hypertension (HT). NT and HT groups were similar with respect to age and basal levels of renin, aldosterone and ANP (34.5 +/- 5.5 in NT and 32.5 +/- 6.3 pg/ml in HT, mean +/- SE). In response to VE, ANP increased to the same extent in both groups (a change of 19.3 +/- 5.2 in NT and of 22.2 +/- 7.1 pg/ml in HT) despite the finding of an exaggerated natriuretic response to VE in essential hypertension (36 +/- 3.5 in NT and 54.9 +/- 6.3 nmol/3 h in HT, P less than 0.02). In addition, the fall in hematocrit and serum protein associated with saline infusion was less marked in HT than NT. The change in ANP induced by VE was inversely correlated with the percent fall in hematocrit and the increment in the fractional excretion of sodium in both groups. These observations suggest that ANPs may participate in the control of the renal response to isotonic VE; however they do not support an unequivocal influence of ANP in the exaggerated natriuretic response to VE of patients with essential hypertension. PMID- 2974247 TI - Synthesis and release of insulinlike growth factor I by mesangial cells in culture. AB - Mesangial cell proliferation is a common hallmark of many glomerular diseases. The exact mechanisms inducing cell proliferation in glomerulosclerosis are not completely understood, and it remains to be determined whether growth factors play a role in this process. Insulinlike growth factor I (IGF I) has been shown to be synthesized in the kidney, and glomerular mesangial cells have receptors for and exhibit mitogenic response to IGF I. We found that mouse glomerular mesangial cells in culture synthesized and released into the culture medium a molecule with immunological and biological features of IGF I. This molecule specifically bound to mesangial cell IGF I receptors; high-pressure liquid chromatographic analysis provided further evidence of its similarity to human recombinant IGF I. Mesangial cells released into the culture medium 6 ng/10(6) cells of IGF I-like material per 24 h in a time-dependent and actinomycin-D inhibitable fashion. These data suggest that IGF I might be locally released by mesangial cells in the glomerulus and act in an autocrine and paracrine fashion. PMID- 2974248 TI - Atrial natriuretic factor and salt adaptation in the teleost fish Gila atraria. AB - It is unknown whether atrial natriuretic factor (ANF) is a mediator of environmental salt tolerance in euryhaline teleost fish. This was investigated in anesthetized Gila atraria, a euryhaline teleost native to springs of pleistocene Lake Bonneville. Plasma levels of immunoreactive (ir) ANF [using anti-human ANF (99-126) antibodies] in fish obtained from a "fresh water" spring were significantly lower (146 +/- 27) than those in fish obtained from a "1% NaCl" spring (347 +/- 21 pg/ml, P less than 0.01). Electron micrographs of fish atrial and ventricular cardiocytes demonstrated many perinuclear granules, which closely resembled ANF-containing secretory granules seen in mammalian atriocytes. Fish heart extract contained ANF-like material of 3 kDa, which caused a marked diuresis and natriuresis in rats. In a second study, fish from a 1% NaCl spring were kept in tanks. One-third of the fish were maintained in 1% NaCl and one third each were either adapted to fresh- or high-salt water. After 12 days, plasma irANF levels in 1% NaCl fish were 343 +/- 55, in fresh water fish 213 +/- 20 and in high-NaCl fish 691 +/- 79 pg/ml. These values differed significantly from each other (P less than 0.01). There was a close correlation between plasma irANF levels and both environmental and internal salt concentration. These data suggest that piscine ANF is an as yet unrecognized mediator of salt tolerance in this teleost and that ANF in these animals closely resembles mammalian ANF. PMID- 2974249 TI - V1 vs. combined V1+V2 vasopressin blockade after hemorrhage in conscious dogs. AB - We examined the hypothesis that V2-like receptors might contribute to the hemodynamic response seen after blockade of the vasoconstrictor (V1) effect of arginine vasopressin (AVP) in nonhypotensive hemorrhage. Seven chronically instrumented dogs were bled 15 ml/kg within 15 min on two different days, at least 3 days apart, and then injected either with the V1 antagonist [1-(beta mercapto-beta,beta-cyclopentamethylenepropionic acid)2-(O-methyl)tyrosine]AVP [d(CH2)5Tyr(Me)AVP, 10 micrograms/kg] or with the combined V1+V2 antagonist [1(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid)2-(O-ethyl)-D tyrosine)4-valine]AVP [d(CH2)5-D-Tyr-(Et)VAVP (10 micrograms/kg)]. Mean arterial pressure, heart rate, and cardiac output (electromagnetic flowmeter) were measured before as well as after hemorrhage and for 10 min after antagonist administration. Both antagonists given after hemorrhage significantly decreased mean arterial pressure as well as total peripheral resistance and increased cardiac output. The V1 antagonist also increased heart rate significantly. No significant hemodynamic changes were measured in another group of six dogs in the absence of antagonist treatment. Although hemodynamic changes tended to be greater with the V1 antagonist than with the combined V1+V2 antagonist, a significant difference between the two analogues was established only for heart rate. These results indicate that in hemorrhage interaction with V2-like receptors plays only a modest role in the hemodynamic changes after V1 blockade in conscious dogs, contrary to what was found in dehydration. PMID- 2974250 TI - Microvascular morphometry and perfusion in renal hypertension-induced cardiac hypertrophy. AB - This study examined alterations in microvascular morphometry and perfusion occurring concomitantly with changes in coronary blood flow (CBF), flow reserves, and coronary vascular resistance (CVR) in myocardial hypertrophy. New Zealand White rabbits (n = 28) with one-kidney, one-clip hypertension (1K,1C) or uninephrectomy (control) were examined 4 wk after surgery in anesthetized open chest preparations. Animals were divided into two experimental series. In the first series, flows were determined with radioactive microspheres during rest and adenosine-induced vasodilation. In a second series, fluorescein isothiocyanate dextran (FITC-dextran) was injected to visualize the perfused arteriolar and capillary beds. The total vasculature was marked with an alkaline phosphatase stain. Mean arterial pressure was elevated in 1K,1C animals (110 +/- 7 mmHg, means +/- SE) when compared with controls (77 + 4 mmHg), and the myocardial weight was greater. Resting CBF was higher in 1K,1C animals compared with controls (227 +/- 21 vs. 168 +/- 12 ml.min-1.100 g-1), and the flow reserve was reduced. Minimal CVR was higher in 1K,1C compared with controls (0.190 +/- 0.035 vs. 0.091 +/- 0.018 mmHg.ml-1.min.100 g). The number of capillaries per squared millimeter was not different from control (2,448 + 121 vs. 2,216 +/- 132/mm2) and the percent perfused was similar (56 +/- 2 vs. 61 +/- 3%). The arteriolar density in cardiac hypertrophy was lower (1.2 +/- 0.2 vs. 2.1 +/- 0.3/mm2), and the percent perfused was higher (86 +/- 5 vs. 63 + 6%) compared with controls. Thus, in myocardial hypertrophy, the anatomical density and volume fraction of arterioles appears to be reduced, and the percentage of arterioles perfused increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974251 TI - Intralipid effect on normal and hypoxic remodeled rat pulmonary vasculature. AB - Intralipid (IL) infusion has been associated with pulmonary vasoconstriction and decreased oxygenation and may worsen preexisting pulmonary vascular changes. To investigate this, we infused IL or 0.9% saline for 1 wk in normal Sprague-Dawley rats and in rats with vascular changes induced by a previous 2-wk exposure to chronic hypobaric hypoxia (air at 380 mmHg). At postmortem we quantitatively evaluated arterial changes in the left lung by light microscopy and alterations in endothelial cells in the right lung by electron microscopy. In rats maintained in room air, 1-wk IL infusion resulted in extension of muscle into alveolar wall and duct arteries (P less than 0.001 for both), medial hypertrophy of arteries 50 99 microns external diameter (P less than 0.01), reduced arterial density (P less than 0.05), and an increase in volume density of endothelial smooth endoplasmic reticulum (P less than 0.05). In post-hypoxia rats, however, IL infusion did not induce further progression of the more severe arterial and endothelial changes observed. To determine whether IL-associated vascular abnormalities may be related to vasoconstriction, different rats were instrumented under pentobarbital sodium anesthesia with indwelling cardiovascular catheters and, 2 days later, with the animals fully conscious, the hemodynamic response assessed. An acute 15 min IL infusion caused a significant increase in pulmonary artery pressure and mild hypoxemia (P less than 0.05 for both) in room air rats but not in the posthypoxia group. This response was not, however, sustained over a 2-day IL infusion. Thus IL induces pulmonary vascular abnormalities that do not appear to be related to vasoconstriction or hypoxemia. We speculate that differences in endothelial metabolism of IL in room air and posthypoxia rats may explain the lack of IL-related abnormalities in the latter group. PMID- 2974252 TI - Atrial natriuretic hormone effect on renal function and aldosterone secretion in sodium depletion. AB - The effects of atrial natriuretic hormone (ANH) on aldosterone secretion and renal function have been well documented, but the physiological role of ANH is still unknown. To address this issue, eight normal men were infused for 4 h with low-dose (1.1 pmol.kg-1.min-1) human [Ser-Tyr28]ANH after 3 days of low-salt (LS) diet. The same subjects were also studied with placebo infusion on LS and high salt (HS) diet. ANH infusion caused doubling of urine flow, a fourfold increase in urinary sodium excretion, and a slight increase in potassium excretion. Immunoreactive ANH levels increased from 3.1 +/- 0.5 to 21.0 +/- 1.9 pmol/l during ANH infusion. ANH infusion suppressed plasma renin activity (PRA) to one third of the basal value, and plasma aldosterone was suppressed from 46.5 +/- 6.5 to 20.9 +/- 2.6 ng/dl. Low-dose ANH infusion caused a marked increase in urine flow and urinary sodium excretion and prominent suppression of PRA and plasma aldosterone in sodium-depleted subjects. These results suggest a physiological significance of ANH in regulation of kidney function and aldosterone secretion. PMID- 2974253 TI - Renal dose response and pharmacokinetics of atrial natriuretic factor in dogs. AB - The present studies investigated the dose-plasma level-response relationships with the use of increasing doses of atrial natriuretic factor [ANF-(99-126)] administered by constant infusion in conscious dogs. The preinfusion plasma immunoreactive ANF increased 12, 19, 23, and 35 times during 45-min consecutive infusions of 50, 75, 125, and 175 ng.kg-1.min-1, respectively. Over this pharmacological range, natriuresis increased linearly with the infused dose (r = 0.99, n = 5) to a maximum response of +1,550%, despite the significant gradual fall of blood pressure, which attained a minimum of 83 mmHg (-26%) at 125 ng.kg 1.min-1. There was no change of glomerular filtration rate (GFR) or renal plasma flow at any dose. A very similar renal response was found in 13 other dogs infused with the highest dose only in which the pharmacokinetic parameters of ANF (99-126) were estimated. Metabolic clearance rate during the infusion was 1.09 +/ 0.19 l/min. The postinfusion decay curve of plasma immunoreactive ANF was best described by a biexponential function. Plasma disappearance half time was 1.44 min during the rapid phase and 10.3 min during the slow phase of elimination. The results show that 1) natriuretic response to ANF in the pharmacological range is dose dependent and occurs despite a pronounced hypotension, 2) increase in GFR is not a prerequisite of ANF-induced natriuresis, and 3) ANF is rapidly eliminated from the circulation, suggesting an intensive uptake and/or degradation in the target tissues. PMID- 2974254 TI - In search of trophoblast-lymphocyte crossreactive (TLX) antigens. PMID- 2974255 TI - Factors that influence cutaneous reactions following administration of thiopentone and atracurium. AB - Atracurium was administered by a variety of techniques to determine whether these influence the onset or duration of muscular relaxation, and the frequency of cutaneous reactions, after a standard induction dose of thiopentone. One-hundred and-fifty patients were allocated randomly to receive the drug by one of five methods: into a fast-flowing crystalloid infusion in the antecubital fossa; into a winged needle in the antecubital fossa with flushing after the thiopentone; into a winged needle in the antecubital fossa without flushing; into a winged needle in the dorsum of the hand without flushing. The above groups received atracurium freshly removed from the refrigerator whereas the fifth group were given atracurium which had been maintained at room temperature for at least 2 weeks. The frequency of cutaneous reactions was between 60 and 70% overall and there were no significant differences either in this or in the onset or duration of action between the groups. A further 25 patients with a history of drug allergy were also investigated by the first method and showed no significant differences in response, but 25 patients aged over 70 years had a significantly lower frequency of cutaneous reactions with a higher frequency of hypotension than the other groups. PMID- 2974256 TI - Hypersensitivity reactions to alfentanil? PMID- 2974257 TI - A hazard associated with removal of carbon dioxide cylinders. PMID- 2974258 TI - Regulation of immune functions by human surfactant. AB - Human peripheral blood mononuclear cells (MNC) were incubated in vitro with highly purified human surfactant to examine its effect on various T cell functions. Surfactant inhibited DNA synthesis by lymphocytes in response to concanavalin A (Con A), phytohemagglutinin (PHA), and in the autologous mixed lymphocyte reaction (AMLR). In contrast, surfactant had no effect on pokeweed mitogen (PWM, T cell-dependent B lymphocyte mitogen)-induced DNA synthesis or on interleukin-2 (IL-2) receptor expression on T cells activated with PHA, Con A or PWM. Furthermore, surfactant had either no effect or enhanced (depending upon the concentration of IL-2 used) the response of exogenous recombinant IL-2 on IL-2 dependent T cell line, In vitro addition of recombinant IL-2 corrected the suppressive effect of surfactant on the AMLR. These data show immunosuppressive effect of surfactant on T lymphocyte functions. PMID- 2974259 TI - [Postoperative analgesia: epidural injection of dexamethasone sodium phosphate]. AB - Epidural administration of steroids has been suggested for the prevention of postoperative epidural fibrosis after lumbar lamino-arthrectomy. In order to assess the efficacy of this technique on pain occurring after such surgery, the demand of pentazocine during the first 24 postoperative hours was studied in 39 patients randomly assigned to two groups. Pain intensity was assessed by a five point verbal scale (0: none; 1: poor; 2: moderate; 3: severe; 4: very severe) at five intervals (0-4, 5-8, 9-12, 13-16 and 17-24 h). The patients in group T (n = 20) did not receive any steroid, whereas those in group C (n = 19) were given, just after the end of surgery, a single dose of dexamethasone (4 mg) via an epidural lumbar catheter previously inserted by the surgeon. In group T, 18 patients required one or several intramuscular injections of pentazocine, whereas only three patients of group C (p less than 0.001) did so. Patients in group T expressed more severe pain (4 moderate, 12 severe, 3 unbearable) than those patients who had received steroids (1 moderate, 1 severe, 1 unbearable). As a consequence, they requested more pain killer (30 injections vs 8 injections, respectively; p less than 0.001) and sooner than patients of the steroid group (8 h vs 12 h; p less than 0.05). It was concluded that epidural administration of dexamethasone was helpful in preventing postoperative pain after lumbar lamino arthrectomy. PMID- 2974261 TI - Reversibility of diaphragm fatigue by mechanical hyperperfusion. AB - Diaphragm function is thought to depend on the balance between diaphragm blood flow and metabolic demand. If blood flow is inadequate, fatigue should ensue. Likewise, in the presence of fatigue, function should improve when blood flow is increased. To test this hypothesis, we evaluated the effect of increasing blood flow to the fatigued diaphragm. Studies were performed on anesthetized, mechanically ventilated dogs in which strips of costal diaphragm were developed in situ. Strip tension was measured with an isometric tension transducer. The inferior phrenic artery supplying the strip was cannulated and pump perfused at a pressure of 92 +/- 3 mm Hg; phrenic artery flow and pressure were continuously monitored with in-line doppler flow probes and pressure transducers, respectively. Fatigue was produced by electrically stimulating strips to contract 15 times/min (initial tension 80% of maximum, duty cycle 50%). Rhythmic contraction resulted in a downward shift in the diaphragm force-frequency relationship in all strips. In eight strips, stepwise increments in phrenic artery perfusion pressure to 161 +/- 8 and 281 +/- 17 mm Hg were produced by increasing pump speed at 6 and 8 min into rhythmic stimulation; in four strips, phrenic artery perfusion pressure was increased to 152 +/- 20 and 257 +/- 32 mm Hg at 20 and 25 min into rhythmic stimulation, respectively. Each increase in phrenic artery pressure resulted in increases in phrenic artery flow and diaphragm tension and produced an upshift in the diaphragm force-frequency relationship.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974260 TI - Cellular influx and activation increase macrophage cytotoxicity and interleukin-1 elaboration during pulmonary inflammation in rats. AB - This study was undertaken to investigate the effects of pulmonary inflammation induced by bacillus Calmette-Guerin (BCG) on the density distribution of lavaged alveolar macrophages. We sought to determine macrophage cytotoxicity and interleukin-1 elaboration in density-defined subpopulations of macrophages during tissue inflammation. At all time points after intravenously administered BCG, lavaged alveolar macrophages contained increased percentages of higher density cells. Alveolar macrophage cytotoxicity against the rat sarcoma cell line XC increased maximally 2 to 6 days after intravenous administration of BCG before declining on Day 13. Macrophage interleukin-1 elaboration increased maximally 14 days after administration of BCG before declining on Day 23. Additionally, macrophage cytotoxicity and interleukin-1 elaboration were increased above normal in cells from each of five density fractions. We conclude that a subpopulation of higher density macrophages, probably recently derived from blood monocytes, accumulates in inflammatory sites. Cellular activation increases the cytotoxicity and interleukin-1 elaboration by macrophages in all density-defined subpopulations and obscures the relationship between cellular density and function that is present in normal animals. PMID- 2974262 TI - Fluid balance in acute and chronic lung disease. PMID- 2974263 TI - A comparison of oral procaterol and albuterol in reversible airflow obstruction. AB - The efficacy and safety of orally administered procaterol hydrochloride, a potent beta 2-adrenergic bronchodilator, was compared with that of albuterol in an eight center, double-blind study conducted in 223 patients with mild to moderate, reversible bronchial airway obstruction. After a 1-wk placebo washout period, patients were administered either procaterol 0.05 mg twice daily for 2 wk followed by 0.10 mg twice daily for 10 wk or albuterol 2 mg three times a day for 2 wk followed by 4 mg three times a day for 10 wk. Spirometry determinations 1.5 h postdose showed consistently greater percent improvements from predose in FVC, FEV1, and FEF25-75 with procaterol than with albuterol at Weeks 1, 2, 4, 8, and 12. Treatment differences were statistically significant (alpha = 0.05) after 2 wk, 2 months, and 3 months of treatment. Bronchodilatation was evident 0.5 h after dosing and peaked at 1.5 to 3 h postdose for both treatments. The duration of action (i.e., time until spirometry determinations were lower than those at 0.5 h postdose) was at least 5 h after procaterol but only 3 h after albuterol. There was no evidence of tolerance with continued procaterol treatment, whereas a diminished duration of response to albuterol was observed with long-term treatment. Tremor was reported statistically more frequently in patients receiving procaterol than in those receiving albuterol (alpha = 0.05); the frequencies of other adverse events were similar for the two groups. No statistically significant treatment differences were noted for asthma symptoms, global evaluations, ECG results, vital signs, or clinical laboratory measurements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974265 TI - [Zinc and the skin]. PMID- 2974264 TI - Mesocortical dopaminergic function and human cognition. AB - In summary, we have reviewed rCBF data in humans that suggest that mesoprefrontal dopaminergic activity is involved in human cognition. In patients with Parkinson's disease and possibly in patients with schizophrenia, prefrontal physiological activation during a cognitive task that appears to depend on prefrontal neural systems correlates positively with cognitive performance on the task and with clinical signs of dopaminergic function. It may be possible in the future to examine prefrontal dopamine metabolism directly during prefrontal cognition using positron emission tomography and tracers such as F-18 DOPA. PMID- 2974266 TI - [Multicenter survey related to the frequency of positive patch tests with mercury and thiomersal]. AB - A multicentric study concerning the frequency of positive allergic patch test reactions to mercury and to thiomersal has been conducted in France and in Belgium among 2,000 adult patients submitted to routine patch testing. 73 (3.6 p. 100) patients had a positive patch test to mercury and 47 (2.3 p. 100) to thiomersal, 22 (1.1 p. 100) reacted positively to both mercurials. These high figures are most probably in relation with a broad use of mercurials in both countries, as antiseptics as well as preservative agents in topical drugs. They lead to a careful use of mercurials, which have to be avoided when they can be advantageously replaced by other antiseptics or preservative agents. As far as cosmetics are concerned, the use of mercurials (chemical nature and concentration) is restricted by a Recommendation of the European Council. PMID- 2974268 TI - [Rosacea: histopathologic study of 75 cases]. AB - Few histological studies have been devoted to rosacea, a common but ill-defined disease. We have examined histological sections obtained from 75 patients (41 men aged from 23 to 72 years and 34 women aged from 32 to 68 years), thereby confirming the diagnosis of rosacea. On average, 4 sections from each patient were examined. Blocks from cases with granuloma were serially sectioned in search of remains of hair follicles, degenerated collagen fibres and Demodex folliculorum. Elastotic degeneration, vasodilatation and the number of hair follicles were evaluated; the nature and disposition of inflammatory infiltrates were determined, and the presence of D. folliculorum was looked for. Elastotic degeneration, varying degrees of vasodilatation and inflammatory infiltrates were found in all patients. The infiltrates were predominantly lympho-histiocytic in 62 cases, neutrophilic around skin appendages in 6 cases, tuberculoid unrelated to appendages in 4 cases and tuberculoid surrounding follicle-related necrosis in 3 cases. The number and appearance of hair follicles seemed to be those normally observed in the regions where our specimens were taken. On the basis of histological, immunological and therapeutic arguments, it has been postulated that D. folliculorum was involved in the pathogenesis of rosacea, notably in its granulomatous form. In particular, several cases of granuloma containing the acarid in its core have been reported. However, the significance of such findings remains uncertain. D. folliculorum might induce the formation of a granuloma, but it is also possible that granulomas are consecutive to the destruction and resorption of hair follicles, the mite being "digested" more slowly than the epithelial structures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974267 TI - [Gram-negative bacteria folliculitis]. AB - Gram-negative folliculitis was first described in 1968 and had since given rise to numerous publications. To our knowledge, no case has yet been published in France, although we observed 6 of them in 12 months. Between March, 1985 and March, 1986, samples of pus for bacteriological examination were obtained from all our patients with acnea vulgaris resistant to the standard treatment (i.e. oral tetracycline combined with topical applications of tretinoin or benzoyl peroxide), thus enabling us to detect patients who had a Gram-negative bacillus (GNB) in at least one pustule. Inflammatory and painful episodes were noted in every case. Tetracyclines, initially effective, gradually lost their activity. The lesions were always limited to the face and consisted of: either superficial small pustules located in the naso-labial line and on the upper lip and chin, associated with inflammatory papulopustular lesions of the cheeks and perioral region; or deeply sited and painful nodules of the cheeks. Various GNBs of the lactose-fermenting Gram-negative rod group were isolated from the superficial lesions, and a Proteus mirabilis strain from the deep lesions. The GNB was never found in all samples but only in 1 to 3 pustules. Four patients were treated with antibiotics proved to be active in vitro against the responsible organisms, and their lesions disappeared within a fortnight. Two patients were given isotretinoin in doses of 1 mg/kg/day with a satisfactory result after 2 and 3 months respectively. The prevalence of Gram-negative folliculitis is probably underestimated. The clinical picture is stereotyped. GNBs are found in some pustules but rarely in all.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974270 TI - [Acitretin (RO 10-1670). A successor of etretinate?]. PMID- 2974269 TI - [Cutaneous allergic accidents caused by insulin. Current aspects apropos of 2 cases]. AB - Current data concerning cutaneous allergy to insulin may be illustrated by the two cases reported here. One was a woman with gestational diabetes; she was treated with bovine insulin and developed generalized urticaria which subsided after switching to human insulin. The other was a woman who had pruritus localized to the site of injection with every type of insulin and in whom laboratory examinations showed an increase of specific IgE. Immunological reactions have been described since the time when exogenous insulin was introduced as a treatment of diabetes. The wide use of purified human insulin has considerably reduced their incidence but benign local and immediate systemic reactions are still being reported, their estimated frequency varying from 10 p. 100 to 50 p. 100 of the patients treated. In reality, allergy to insulin itself is extremely rare compared with allergic reactions to preservatives, such as metacresol, additives (protamine and zinc and contaminants present in insulin preparations: desamido-insulin. True allergic reactions to insulin may be localized or generalized and biphasic, and in most cases they are IgE-mediated. Some late local reactions, as well as atrophy, can be ascribed to delayed hypersensitivity. Treatment includes: (i) change in the type of insulin used; (ii) systemic or topical corticosteroid therapy; (iii) antihistamines and aspirin, and (iv) desensitization. The allergic complications of insulin therapy are benign; they usually do not require any particular treatment and often spontaneously regress. PMID- 2974271 TI - [Use of methotrexate in a child with trisomy 21. Value of blood levels]. PMID- 2974272 TI - [The uretheral syndrome in men or prostatodynia]. AB - Based on the results of the clinical and urodynamic investigation of 15 patients, presenting with functional genitourinary symptoms, the authors report the efficacy of treatment with alpha-blockers. They stress the importance of the clinical history and of static sphincterometry in this type of patient. PMID- 2974273 TI - Stabilities of lyophilized Staphylococcus aureus typing bacteriophages. AB - Staphylococcus aureus bacteriophages (25 phages) were lyophilized in aliquots 12 to 18 years ago and stored in vacuo at -20 degrees C. Eight viruses each lost one log titer, while seventeen retained the original titers. The use of lyophilized phages provided more reproducible phage typing and reduced by 75% the complexity and cost. This important test is thus made feasible for more laboratories. PMID- 2974274 TI - Penicillin allergy--a rare paediatric condition? AB - A total of 298 children with a history of adverse reactions in connection with oral penicillin treatment were investigated with a radioallergosorbent test for penicillin metabolites, the skin prick test, and oral challenge with penicillin V. No severe reactions were seen. In 30 (10%) of the subjects slight to moderate skin reactions were observed on the seventh to 10th day of the challenge period. Between one to four years after the oral challenge 222 children were reinvestigated by interview. One hundred and ten had been given treatment by penicillin and 103 (94%) of these children tolerated the new treatment well and without any adverse reactions. We conclude that the term 'penicillin allergy' is often misused. Such a diagnosis should be established by clinical investigation. PMID- 2974275 TI - Biological functions of the NS1 protein of an influenza B virus mutant which has a long carboxyl terminal deletion. AB - To clarify the function of the NS gene of a highly cytolytic mutant of influenza virus B/Yamagata/1/73 which expresses an NS1 protein with a long carboxyl terminal deletion (clone 201), we prepared a single gene reassortant (201 L-77) and a control reassortant (YL-20) in which all the genes were of wild type influenza virus B/Lee/40 origin except NS gene which was derived from either clone 201 or wild type B/Yamagata. Comparative studies have revealed that 201 L 77 destructed infected cells more severely and much earlier after infection than did YL-20, although both produced comparable amount of infectious virus. The highly cytolytic reassortant 201 L-77 produced a small plaque, while the weakly cytolytic reassortant YL-20 produced a large plaque in MDCK cells. There was little difference between the two reassortants in the time course and the amount of synthesis of viral proteins within the infected cells. However, the mode of synthesis of viral RNA (vRNA) by 201 L-77 was greatly altered compared with YL 20. PMID- 2974276 TI - Adenovirus transcriptional complexes contain EIa encoded tumour antigens physically bound to cellular proteins. AB - Adenovirus type 12 transcriptional complexes were isolated from cells during the early phase of infection. Sedimentation analysis identified a fast sedimenting complex type I and a slow sedimenting complex type II. Both complexes made virus specific RNA complementary to all the early genes and both contained viral DNA, which in type II but not in type I had nucleosome like configuration. Analysis of the proteins of the complexes with antiserum against Ad 12 EIa-beta-galactosidase fusion protein expressed in E. coli demonstrated the following: (a) type I complex contained EIa 45 K protein, which co-precipitated with cellular proteins of mol. wt. 42, 58, and 60 K, (b) type II complex contained EIa 47 K protein, which co-precipitated with major cellular proteins of 35, 40-46 K and minor proteins of 58, 60, 68, 76, 86, and 120-150 K. Association of EIa specific and cellular proteins to transcriptional complexes was sensitive to both 1 M NaCl and DNAse I indicating the DNA binding nature of these proteins. Treatment of transcriptional complexes with 1 M NaCl or DNase I released EIa proteins, which still remained strongly bound to cellular proteins. These findings suggested that EIa proteins bind to viral DNA and that this binding is probably mediated by cellular proteins. PMID- 2974277 TI - Muscle afferent activity and its central projection in man. AB - Muscle is a complex sensory organ as well as a contractile apparatus, and disease processes that produce muscle weakness and wasting will affect the sensory information transmitted by receptors in muscle. In addition, one of the sensory structures in muscle, the muscle spindle, receives a motor innervation, the gamma efferent or fusimotor system, with which the brain can alter the feedback that it receives from muscle. Muscle spindle activity forms the afferent limb of spinal reflexes, such as the tendon jerk, and long-loop reflexes that traverse supraspinal reflex pathways. Muscle spindle activity constitutes the major afferent cue for kinaesthetic sensations and contributes to updating the centrally generated programme for movement. This paper reviews briefly some aspects of muscle spindle activity and its fusimotor control as studied in human subjects using microneurography. PMID- 2974278 TI - Characterization of a myopathy caused by prostaglandin dysfunction. AB - Administration of inhibitors of prostaglandin synthetase to chicken embryos produced myopathies in their skeletal muscles which were characterized by ringbinden, loss of Z-discs, M-bands, and thick and thin filaments and decreased myoblast proliferation and type 2 myotube formation. The effect of administration of prostaglandins on myoblast proliferation was also examined and PGE was found to suppress proliferation. There was also a tendency for PGF2 alpha to suppress and PGI2 to stimulate proliferation, although neither of these effects were statistically significant. PGA, PGB and PGD did not affect myoblast proliferation. PMID- 2974281 TI - Subacute effects of propranolol and B 24/76 on isoproterenol-induced rat heart hypertrophy in correlation with blood pressure. AB - We compared the potential beta-receptor blocker, B 24/76 i.e. 1-(2,4 dichlorophenoxy)-3[2-3,4-dimethoxyphenyl)ethanolamino]-prop an-2-ol, which is characterized by beta 1-adrenoceptor blocking and beta 2-adrenoceptor stimulating properties with propranolol. The studies were performed using an experimental model of isoproterenol-induced heart hypertrophy in rats. A correlation of the blood pressure was neither found in the development nor in the attempt to suppress the development of heart hypertrophy with the two beta-receptor blockers. Both beta-blockers influenced the development of hypertrophy to a different, but not reproducible extent. It was possible to suppress the increased ornithine decarboxylase activity with both beta-blockers in hypertrophied hearts, but there was no effect on the heart mass. Neither propranolol nor B 24/76 could stop the changes in the characteristic myosin isoenzyme pattern of the hypertrophied rat heart. Thus, the investigations did not provide any evidence that the beta-receptor blockers propranolol and B 24/76 have the potency to prevent isoproterenol from producing heart hypertrophy. PMID- 2974280 TI - Atrial-natriuretic-factor mRNA is developmentally regulated in heart ventricles and actively expressed in cultured ventricular cardiac muscle cells of rat and human. AB - Atrial-natriuretic-factor (ANF) mRNA is actively expressed in ventricular heart myocytes of neonatal rats and fetal and young (2.5-year-old) humans. In rat, ANF mRNA transcription is repressed by the 23rd day of postnatal development, and in human it is no longer detected by the 14th year of development. Its expression is observed in atrial myocytes of both of these species at all ages. ANF mRNA is expressed in primary cultures of ventricular cardiac muscle cells prepared from both neonatal-rat and fetal-human hearts. Surprisingly it is also very actively expressed in cultures of adult rat ventricular cardiac muscle cells. The effect of several hormones on the expression of ANF mRNA in rat and human myocyte cultures was evaluated. These studies demonstrate that ANF mRNA transcription is developmentally regulated in both rat and human heart ventricles and suggest that ventricular cardiac muscle-cell cultures may be useful in studying the regulation of the expression of this gene. PMID- 2974279 TI - Alpha 2-antiplasmin Enschede is not an inhibitor, but a substrate, of plasmin. AB - alpha 2-Antiplasmin Enschede is a variant of alpha 2-antiplasmin which has lost its ability to inhibit plasmin irreversibly and which is associated with a haemorrhagic disorder [Kluft et al. (1987) J. Clin. Invest. 80, 1391-1400]. The abnormal protein was purified from the plasma of a homozygous patient and subjected to one-dimensional peptide mapping using papain for digestion. A slightly abnormally migrating polypeptide (Mr 17,000) was found which represented the C-terminal part of the molecule (the N-terminus of the polypeptide corresponded to Gly-338 in normal alpha 2-antiplasmin) and which contained the reactive centre. The interaction of plasmin with alpha 2-antiplasmin Enschede was studied by adding plasmin to plasma of the homozygous patient. SDS/polyacrylamide gel electrophoresis and immunoblotting showed that no complex persisted, but that the abnormal alpha 2-antiplasmin was cleaved into two fragments of Mr 56,000 and 14,000 respectively. The latter fragment co-migrated with the post-complex peptide, which is cleaved from normal alpha 2-antiplasmin during complex formation with plasmin. In a purified system, catalytic amounts of plasmin rapidly cleaved alpha 2-antiplasmin Enschede into the aforementioned fragments. In kinetic studies alpha 2-antiplasmin Enschede reversibly and temporarily inhibited the plasmin-catalysed hydrolysis of D-valyl-L-leucyl-L-lysine p nitroanilide ('S-2251') as a competitive inhibitor (Ki,app. 35 nM). It was concluded that alpha 2-antiplasmin Enschede apparently forms a normal complex with plasmin. The complex is, however, not stable, but disintegrates rapidly to a cleaved form of alpha 2-antiplasmin Enschede and active plasmin. The abnormal protein thus behaves like a substrate, instead of an inhibitor, of plasmin. PMID- 2974282 TI - The use of separated subunits of yeast phosphofructokinase for the isolation of subunit-specific antibodies from polyclonal antisera. AB - The subunits alpha and beta of yeast phosphofructokinase have been separated under denaturing conditions by ion exchange chromatography and then separately immobilized on Sepharose 6MB. The resulting affinity gels were used to fractionate polyclonal phosphofructokinase antisera from rabbit into subunit specific antibodies. In addition to subunit-specific antibodies also antibody fractions which recognize both types of subunits of phosphofructokinase could be desorbed from the affinity gels. Antibodies directed either to the alpha- or to the beta-subunits were found to inhibit the enzyme activity. The results are discussed in terms of a partial identity of the alpha- and beta-chain of phosphofructokinase. PMID- 2974283 TI - Mannose 6-phosphate increases the affinity of its cation-independent receptor for insulin-like growth factor II by displacing inhibitory endogenous ligands. AB - The insulin-like growth factor II (IGF-II) and glycoprotein lysosomal enzymes containing mannose 6-phosphate (M6P) bind with high affinity to two separate sites on the same receptor molecule (Morgan et al. Nature 329:301). The addition of free M6P significantly increases the affinity of some preparations of the M6P/IGF-II receptor (M6P/IGF-II-R) for IGF-II. We conducted this study to test the hypothesis that this effect is the result of displacement of M6P-related ligands that inhibit IGF-II binding. First we found that although M6P caused a 66% increase in the binding of IGF-II to microsomes prepared from IM9 cells, it had no effect, under identical conditions, on binding to receptor on the surface of intact cells. Secondly, extensive washing of rat liver microsomes in the presence of 3 mM M6P, followed by removal of the M6P by further washing, abolished the effect by raising binding to levels seen in the presence of M6P. M6P, then, had no additional effect. Finally, when IGF-II-affinity purified receptor was repurified by ultracentrifugation on a sucrose gradient, binding to the pure receptor peak was not affected by M6P. We conclude that there is no intrinsic positive cooperativity between free M6P and the IGF-II-binding site of the M6P/IGF-II-R. The reported M6P-induced increase in IGF-II binding appears to be due to the displacement of contaminating inhibitory endogenous ligands. PMID- 2974284 TI - Activation of muscle phosphofructokinase by alpha-glucose 1,6-bisphosphate and fructose 2,6-bisphosphate is differently affected by other allosteric effectors and by pH. AB - Citrate, ATP and AMP affect similarly the activation of muscle phosphofructokinase by alpha-glucose 1,6-bisphosphate and fructose 1,6 bisphosphate, but they affect differently its activation by fructose 2,6 bisphosphate. Activation by alpha-glucose 1,6-bisphosphate and fructose 2,6 bisphosphate is also differently affected by pH. This suggest that both alpha glucose 1,6-bisphosphate and fructose 1,6-bisphosphate induce the same conformational change on muscle phosphofructokinase, distinct from that produced by fructose 2,6-bisphosphate. PMID- 2974285 TI - Isolation of a biologically active fragment from the carboxy terminus of the fetal rat binding protein for insulin-like growth factors. AB - We have purified a 14 kDa fragment of the 30 kDa binding protein for insulin-like growth factors (IGFs) from BRL-3A cell conditioned medium. The fragment binds IGF I and IGF-II with similar specificity to the 30 kDa binding protein, but with lower affinity. It corresponds to the carboxy terminus of the native binding protein (residues 148-270), and is thought to arise by proteolysis. We infer that this region of the native binding protein contains, at least in part, the IGF binding domain. PMID- 2974286 TI - The affinities of prostaglandin H2 and thromboxane A2 for their receptor are similar in washed human platelets. AB - Both thromboxane A2 (TXA2) and its precursor prostaglandin H2 (PGH2) are labile and share a common receptor. The affinities of these two compounds for their putative common receptor are unknown. We compared the potencies of TXA2 and PGH2 to aggregate human platelets and bind to the TXA2/PGH2 receptor. TXA2 was more potent than PGH2 in initiating aggregation in platelet-rich plasma, EC50 of 66 +/ 15 nM and 2.5 +/- 1.3 microM, respectively. In washed platelets, however, PGH2 was more potent than TXA2 with EC50 values of 45 +/- 2 nM and 163 +/- 21 nM, respectively. The affinity of these two compounds in washed platelets was determined in radioligand competition binding assays employing [125I]-PTA-OH. The Kd values for PGH2 and TXA2 were 43 nM and 125 nM, respectively. The results demonstrate that the affinity of PGH2 for the platelet TXA2/PGH2 receptor is greater than previously thought. The data raise the possibility that PGH2 may significantly contribute to the responses attributed to TXA2 in vivo. PMID- 2974287 TI - Two homologous oncogenes, HST1 and INT2, are closely located in human genome. AB - Pulsed field gel electrophoresis and Southern blot analysis showed that the human oncogenes, HST1 and INT2, which code for proteins homologous to fibroblast growth factors, are less than 45 kb apart on the long arm of chromosome 11. Moreover, analysis of two overlapping cosmid clones, one containing INT2 and the other HST1 sequences, showed that HST1 is located about 35 kb downstream of INT2 in the same transcriptional orientation. The observed close proximity of the INT2 and HST1 genes may provide important insight on the origin and regulation of expression of these related genes. PMID- 2974288 TI - Employee counseling: a new subspecialty for the psychiatric-mental health clinical nurse specialist. PMID- 2974289 TI - Purification of a specific peptidase in bovine atria for the processing of pro atrial natriuretic factor. AB - A serine protease which catalyses the conversion of the precursor of ANF (atrial natriuretic factor) to the active circulating form, ANF-(99-126)-octacosapeptide, was purified from a particulate fraction of bovine atria. The enzyme was solubilized with a buffer containing 1.6M KCl. The molecular mass of the purified enzyme was 580 kDa on gel filtration, whereas on sodium dodecyl sulfate polyacrylamide gel electrophoresis a cluster of six bands with molecular masses around 30 kDa was observed. The purified enzyme produced the ANF-(99-126) octacosapeptide from partially purified bovine pro-ANF by the selective cleavage of the arginyl peptide bond in the -Pro97-Arg98-Ser99-sequence in pro-ANF. It is likely that the enzyme selectively cleaves the Arg98-Ser99 peptide bond in pro ANF when ANF is secreted into the circulation. PMID- 2974290 TI - Treatment of prostatic cancer by monthly injections of an LHRH-analogue depot. AB - Thirty-six patients with advanced prostatic cancer were treated by monthly depot injections of a luteinizing-hormone releasing hormone analogue (LHRH-a). Five of these patients were also pretreated for 14 days with cyproterone acetate (CPA) in order to counteract initial increase in testosterone concentration. Two weeks after the initial depot injection the serum testosterone had been reduced to and was maintained at castrate level. Luteinizing hormone and follicle stimulating hormone were also significantly reduced. Of the 31 patients 23 showed objective regression at 3 months, 9 had stable disease and none showed progression. At 3 months 22 patients reported subjective improvement. At 12 months 18 showed objective regression, 7 had withdrawn from therapy and 6 showed progression. Side effects were acceptable and comparable to those following surgical castration. It is shown that CPA counteracts the initial increase in testosterone concentration at initiation of LHRH-a treatment. We conclude that depot preparations of LHRH analogues, both with and without pretreatment with CPA, are useful in the treatment of patients with advanced prostatic cancer. PMID- 2974291 TI - Phase I clinical trial of oral menogaril administered on three consecutive days. AB - Eighteen adult patients with solid tumors were treated with oral menogaril, a new anthracycline antibiotic active against human breast cancer after intravenous administration. The drug was given orally on 3 consecutive days every 4 weeks at doses ranging from 50 to 175 mg/m2/day. Reversible and dose-related leukopenia was the dose-limiting toxicity. At doses from 50 to 150 mg/m2/day, non hematologic side effects of oral menogaril were unfrequent and mild and consisted of nausea and vomiting (1 patient), alopecia (2 patients), mucositis (2 patients) and liver function test abnormalities (3 patients). The only patient treated at a daily dose of 175 mg/m2 developed grade IV leukothrombocytopenia, with fever and gastrointestinal bleeding. This was followed by heart insufficiency and the patient died from multisystem organ failure. A dose of 150 mg/m2/day for 3 consecutive days is recommended for phase II trials with oral menogaril. PMID- 2974292 TI - Uptake of a fluorinated bisphosphonate by cultured bones. AB - The uptake of bisphosphonates into bone was studied using 19-day-old fetal rat bones cultured with a new fluorinated bisphosphonate, difluoromethylidene bisphosphonate (F2MBP). F2MBP uptake was assessed by determining the weight percent of fluoride using electron probe microanalysis. By 30 min the weight percent of fluoride was significantly greater in the F2MBP-treated bones than in controls and continually increased throughout the duration of the experiment to reach a fluoride concentration 6-fold greater than controls after 120 h of incubation. When the peripheral cortical bone was analyzed separately from the interior trabecular bone in the F2MBP-treated bones, the fluoride concentration in the periphery increased until 24 h and then remained somewhat constant, while the interior, which is more actively remodeling, showed a continual increase. The uptake of F2MBP during the 1 to 6 h time intervals demonstrated no differences between vital and devitalized bone and, thus, is not cell-mediated. Because analysis of free fluoride in F2MBP media incubated with bones showed that the concentration of fluoride was less than 1% of the total amount of fluoride, the fluoride detected by the probe was most likely that of the intact molecule and not free fluoride. The rapid uptake of the F2MBP molecule was supported by assessing the effects of short-term F2MBP treatment on subsequent bone resorption, as determined by the release of 45Ca from prelabeled bones. Bones treated with F2MBP for only 5 min exhibited reductions in the percentage of 45Ca released during the remainder of the 120 h incubation period similar to that when F2MBP was continuously in the medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974294 TI - Specialisation in dentistry. PMID- 2974293 TI - Characterization of a (Ca2+ + Mg2+)-ATPase system in the osteoblast plasma membrane. AB - A high affinity, calmodulin-sensitive (Ca2 + Mg2+)-ATPase was demonstrated in the plasma membrane preparation of three different osteosarcoma cell lines previously demonstrated to respond to parathyroid hormone with an increase in cytosolic calcium and a decrease in pH. The maximal velocity of the enzyme activity in the membrane preparations ranged from 0.83 to 2.42 nmol Pi released per min per mg protein with half-saturation constants of 26 nM of free Ca. The enzyme activity was not affected by Na+, K+, ouabain and azide, and exhibited an absolute requirement for Mg2+ ions. These results suggest a possible role for a membrane Ca2 + Mg2+-ATPase in initiating and perpetuating the ionic control of osteoblastic function. PMID- 2974295 TI - Dentistry's role in the AIDS epidemic. PMID- 2974296 TI - Progress report on Surgery V. PMID- 2974298 TI - Transient neonatal tricuspid regurgitation: a Doppler echocardiographic study of three cases. AB - Three patients with normal hearts and no pulmonary abnormality had neonatal tricuspid regurgitation causing cardiorespiratory distress and cyanosis. The signs of tricuspid regurgitation resolved over a few weeks. In the acute phase echocardiography showed gross dilatation of the right atrium and ventricle. The interatrial septum bulged into the left atrium during the whole cardiac cycle. Doppler echocardiography showed clinically significant tricuspid regurgitation, a right to left shunt through the foramen ovale, reduced flow through the pulmonary valve, and in two patients ductal flow into the pulmonary artery. In one patient tricuspid regurgitation was so great that it impeded the opening of the pulmonary valve and produced functional "atresia" of the pulmonary valve. The presence of regurgitant blood flow through the pulmonary valve showed that the "atresia" was functional rather than organic. Doppler echocardiographic study is useful in distinguishing functional neonatal tricuspid regurgitation from structural abnormality of the tricuspid valve. PMID- 2974297 TI - Coronary angioplasty in unstable angina and stable angina: a comparison of success and complications. AB - One hundred and five patients with unstable angina and 175 with chronic stable angina were treated by primary percutaneous transluminal coronary angioplasty. Patients with unstable angina had had symptoms for a shorter time and were more likely to have angiographically complex lesions and lesions less than 10 mm in length than patients with chronic stable angina. Other baseline variables were not significantly different in the two groups. The overall primary success rate was similar in both groups (87% v 86%). Nine of the 14 unsuccessful procedures in those with unstable angina and nine of the 24 unsuccessful procedures in those with stable angina were the result of acute occlusion. These results led to a 9% frequency of procedure related myocardial infarction in patients with unstable angina and a 5% rate in those with stable angina (NS). The procedure related infarct rate tended to be higher in patients with unstable angina who had coronary angioplasty soon after an episode of unstable angina (mean 10 days) than in those in whom it was delayed (mean 35 days) (12% v 3%) (NS). In patients with unstable angina who had had a previous myocardial infarction procedure related infarction was significantly more common (18%) than in patients with no previous myocardial infarction (3%). The difference between those with and without previous infarction was also significant in patients with stable angina (10% v 3%). PMID- 2974300 TI - Endovascular treatment of vertebral arteriovenous fistulas in twenty-two patients. AB - Twenty-two cases of vertebral arteriovenous fistulas treated by embolization are reported. Although such fistulas are usually asymptomatic they may be occasionally responsible for tinnitus. Neurologic complications are exceptional. Spontaneous fistulas have been found predominantly in the C1-C2 region (9 of 13 cases), while post-traumatic fistulas were located in the C5-C6 region and were often iatrogenic in origin (7 of 9 cases). In 17 cases embolization caused definitive closure of the fistula, while the vertebral artery remained patent. In three patients it was also necessary to close the vertebral artery in order to occlude the fistula. In two instances, endovascular management failed to obliterate the fistula. At present, endovascular treatment is the simplest and one of the least aggressive therapeutic approaches to vertebral arteriovenous fistulas. PMID- 2974299 TI - Pharmacokinetics of intraperitoneal teicoplanin in patients with chronic renal failure on continuous ambulatory peritoneal dialysis. AB - The pharmacokinetic profile of teicoplanin, a new glycopeptide antibiotic active against Gram-positive aerobic and anaerobic bacteria, is described in five patients with end-stage renal disease on continuous ambulatory peritoneal dialysis (CAPD). A single 3 mg kg-1 dose was given intraperitoneally in the dialysate during a 6 h dwell time. The drug appeared in the plasma within 15 min at 1.00-0.28 mg l-1 (mean +/- s.d. = 0.70 +/- 0.45) in all five subjects, and peak serum concentrations ranged from 5.53 to 2.80 mg l-1 (4.84 +/- 1.43) at 6 h. Approximately 70% (71 +/- 12) of teicoplanin was absorbed from the peritoneal dialysis fluid during a single 6 h dwell time. The rate constant for peritoneal transfer (lambda d) averaged 0.318 h-1 and the half-life (t1/2 lambda d) was 2.18 h. Further values were serum elimination half-life 114-173 h; total body clearance 263-532 ml h-1; steady-state volume of distribution 68-93 l. This drug profile closely agrees with data reported after intravenous injection in patients on CAPD and suggests that teicoplanin has bidirectional exchange characteristics through the peritoneal membrane, although transfer from the systemic circulation to peritoneal fluid is consistently low. Instillation of teicoplanin in CAPD fluid may be a useful route of administration for treatment of peritonitis and exit site infections in CAPD patients. PMID- 2974301 TI - Obturator bypass: a sixteen year experience with 55 cases. AB - Fifty-five obturator bypasses were performed over a 16 year period for limb salvage. The indications for the choice of this technique included arterial infection, aneurysm formation or soft tissue skin loss at the groin (26 patients), and extensive atherosclerotic disease of the iliofemoral vessels, precluding their use for inflow or outflow (27 patients). There were five perioperative deaths and the five-year patency rate was 37%. Long-term results after above-knee reconstruction substantially exceeded those for below-knee anastomoses (three year patency rates of 71% and 45% respectively). Due to its morbidity and mortality the decision to perform an obturator bypass should be based upon sound indications such as deep infection at the groin, or extensive multilevel arterial disease. Its usefulness in extensive multilevel arterial disease is not demonstrated. PMID- 2974302 TI - Intraoperative evaluation of PTFE, Dacron and autogenous vein as carotid patch materials. AB - Between 1983 and 1986 we prospectively randomized 71 consecutive carotid endarterectomy patients to receive patch closure with one of three materials. Autogenous saphenous vein was used in 18 cases, Dacron velour (Sauvage Filamentous) in 30 cases, and PTFE (Gore-Tex CV patch) was used in 23 cases. Blood loss in the excess of 300 ml was seen in 43% (10/23) of the PTFE group in contrast to only 22% (5/23) of the ASV group and 17% (4/23) of the Dacron group. Persistent suture hole bleeding requiring the use of a significantly larger amount of oxidized cellulose (p less than .001) was noted in the PTFE group. Operating time from clamp release to end of procedure was significantly greater in the PTFE group (p less than .05), reflecting the delay associated with suture hole bleeding. In addition, the handling characteristics of PTFE were inferior to those of Dacron or autogenous saphenous vein. Needle penetration was relatively difficult. PTFE was thicker, less flexible, and a less satisfactory match to the endarterectomized carotid than the other materials. PMID- 2974303 TI - Duodenal obstruction following abdominal aortic reconstruction. AB - Four patients developed duodenal obstruction after 161 abdominal aortic reconstructions, an incidence (2.5%) rivaling that of graft infection and arterioenteric fistula. The diagnosis is easily confirmed by gastrointestinal contrast studies. Duodenal obstruction is usually caused by perigraft collagenous adhesions and is probably less likely to occur if the mobilized duodenum is not replaced directly over the aorta during resuture of the retroperitoneum. Undetected duodenal obstruction leads to rapid dehydration and electrolyte and caloric depletion. PMID- 2974304 TI - Clinical markers of androgenicity in acne vulgaris. AB - Androgenic stimulation of sebaceous glands is necessary for development of acne. If hyperandrogenaemia were a major determinant of acne in women, the frequency of other clinical markers of androgenicity should increase with acne severity. To investigate this, 268 female subjects (aged 12-44 years) were studied. Subjects were divided into groups on the basis of acne severity: physiological, moderate, and severe. With exclusion of women taking oral contraceptives or anti-androgen therapy, subjects in each group were similar with respect to age at menarche and incidence of menstrual irregularity of amenorrhoea. Reports of excessive body hair, and clinical hirsutes on examination were few and there were no significant differences between acne severity groups. No correlation was observed between acne and hirsutes grades in all subjects (rank correlation coefficient = 0.096). Mild male pattern androgenic alopecia occurred in similar proportions of subjects in the three groups. Female pattern androgenic alopecia was observed in only two subjects. We have shown no correlation between acne severity and clinical markers of androgenicity in women. This suggests that in most cases factors other than hyperandrogenaemia are necessary for the development of acne. PMID- 2974305 TI - Papillon-Lefevre syndrome: a study of the long-term clinical course of recurrent pyogenic infections and the effects of etretinate treatment. AB - A family with a clinical variant of Papillon-Lefevre syndrome (PLS), associated with recurrent pyogenic infections, has been followed up for more than 20 years. Of the five living siblings, four were treated with etretinate for a period of at least 21 consecutive months. Clinical follow-up showed that the course of recurrent infections in susceptible PLS patients, although usually more severe in childhood, can be variable and unpredictable. The etretinate therapy resulted in marked improvement of the keratodermas, and was associated with complete remission of the pyodermas on both keratotic and non-keratotic skin. It is, therefore, suggested that etretinate may have a primary role in the prevention of recurrent pyogenic infections in susceptible PLS patients. PMID- 2974306 TI - Acitretin and etretinate in the treatment of palmoplantar pustulosis: a double blind comparative trial. AB - Sixty patients with palmoplantar pustulosis were treated in a double-blind trial with either acitretin (etretin, Ro 10-1670) or with etretinate. The study consisted of 4 weeks of therapy with three 10 mg capsules/day followed by 8 weeks of therapy with a varying number of capsules given daily according to therapeutic response. At the end of the 12-week treatment period, the mean number of pustules (+/- SEM) had decreased from 57.8 (+/- 8.6) to 3.9 (+/- 1.6) in the acitretin group and from 57.1 (+/- 14.1) to 5.7 (+/- 2.7) in the etretinate group. With regard to influence on erythema, infiltration, scaling, and area involved, similar improvements were obtained in both treatment groups. Adverse reactions of the hypervitaminosis A type were observed with almost the same frequency and severity in both treatment groups. The mean number of 10 mg capsules used daily was comparable in the two groups: 2.82 (range 1.23-4.67) for acitretin and 2.77 (range 1.60-4.82) for etretinate. It can be concluded that acitretin and etretinate do not significantly differ with regard to efficacy and overall safety in the treatment of patients with palmoplantar pustulosis. PMID- 2974307 TI - Orthodontic bonding adhesives. PMID- 2974308 TI - [The role of Ca2+-ATpase and its hydrophobic component in the release of Ca2+ from skeletal muscle sarcoplasmic reticulum]. AB - The Ca2+ permeability of proteoliposomes containing Ca2+-ATPase of sarcoplasmic reticulum and its hydrophobic fragment was investigated, using the method of synthetic penetrant ions and the radioisotopic method. The former method was used to determine the diffusional membrane potential formed by Ca2+ concentration gradient. It was demonstrated that Ca2+-ATPase, whose active center is oriented outside, has and asymmetric conductivity, i. e., it facilitates the rapid efflux of Ca2+ from proteoliposomes. This efflux is stimulated by the membrane potential positive inside. The hydrophobic fragment of Ca2+-ATPase forms a Ca2+-channel with a high conductivity for Ca2+. This channel is responsible for the Ca2+ efflux from sarcoplasmic reticulum. PMID- 2974309 TI - Malignancy-associated hypercalcaemia: relationship between mechanisms of hypercalcaemia and response to antihypercalcaemic therapy. AB - The pathophysiological mechanisms of hypercalcaemia were assessed in 50 rehydrated patients with cancer-associated hypercalcaemia. Surprisingly, renal tubular calcium reabsorption appeared to increase progressively as serum calcium rose, suggesting that the nomogram used for the calculation may have been inaccurate, in absolute terms, probably due to its failure to take account of the levels of urinary sodium excretion. There were significant differences in the mechanisms of hypercalcaemia in different patient subgroups, however, independent of differences in urinary sodium excretion. In those with few or no bone metastases, increased renal tubular calcium reabsorption was the principal cause of hypercalcaemia, often in association with increased bone resorption. These abnormalities were thought to reflect the renal and skeletal actions of a tumour associated humoral mediator. The main cause of hypercalcaemia in those with extensive metastatic bone disease was increased bone resorption, with contributions from impairment of glomerular filtration rate and, to a minor extent, increased renal tubular calcium reabsorption. These abnormalities were thought to reflect a mainly local-osteolytic mechanism of hypercalcaemia with secondary impairment of GFR. Of all the biochemical variables assessed pre treatment, the renal tubular component of hypercalcaemia correlated most strongly with post-treatment serum calcium values (r = 0.61, P less than 0.001). Because of their generally lower levels of renal tubular calcium reabsorption, patients with extensive skeletal metastases also had significantly lower post treatment calcium values than patients with few or no metastases (P less than 0.05). These data indicate that the pathophysiological mechanisms of hypercalcaemia are a major determinant of the calcium lowering response after antihypercalcaemic treatment. This should be taken into account during comparative studies of antihypercalcaemic therapy in patients with malignancy. PMID- 2974310 TI - Screening for Down's syndrome. PMID- 2974311 TI - Hepatitis B and Down's syndrome. PMID- 2974312 TI - Participation of serotonergic mechanisms in the anticonvulsant effect of stress in mice treated with pentylenetetrazol. AB - This study analyzed the effects of p-chlorophenylalanine (PCPA) and methysergide pretreatments on prolongation of latency to pentylenetetrazol (PTZ)-induced convulsions in mice submitted to cold/restraint or swim stress. Both anti-5HT agents shortened convulsion latency in non-stressed animals. The effects of cold/restraint and swim stress were markedly suppressed by methysergide. PCPA did not modify the effect of cold/restraint but tended to antagonize that of swim stress. Serotonergic mechanisms possibly exert a tonic inhibitory influence on PTZ-induced convulsions and are important in the manifestation of prolongation of convulsive latency determined by swimming and, possibly, cold/restraint stress. PMID- 2974313 TI - Value of HLA-B27 in the routine screening of patients with back pain. PMID- 2974314 TI - Analysis of combined back clinic outcome. PMID- 2974315 TI - AIDS patients wins case against hospital dental department. PMID- 2974316 TI - Why isn't the hepatitis B vaccine being used? PMID- 2974317 TI - Sexuality education for special needs adolescents. PMID- 2974318 TI - Abnormal expression of the MOS proto-oncogene in human thyroid medullary carcinoma. AB - We have been studying the expression of a range of proto-oncogenes in human thyroid tumour tissue by using Northern blot analysis. We have demonstrated the expression of a MOS mRNA of 1 kb in all thyroid samples. Furthermore, in a medullary carcinoma sample we also observed additional mRNA species of 1.7 and 2.2 kb. Southern blot analysis of DNA prepared from the same tumour sample did not reveal a rearrangement of the gene. These findings are the first report of MOS expression in any human tissue, and indicate that MOS oncogene activation might be important in the development of some thyroid tumours. PMID- 2974319 TI - [Aortic insufficiency in Down's syndrome: a color Doppler echocardiography study]. PMID- 2974320 TI - [Transluminal coronary angioplasty performed by percutaneous brachial approach]. PMID- 2974321 TI - A point mutation in the extracellular domain of the human CSF-1 receptor (c-fms proto-oncogene product) activates its transforming potential. AB - A human CSF-1 receptor containing an "activating" mutation in its extracellular domain (serine for leucine 301) induced morphologic transformation, anchorage independent growth, and tumorigenicity in mouse NIH 3T3 cells. A second regulatory mutation within the receptor's intracytoplasmic carboxy-terminal tail (phenylalanine for tyrosine 969) augmented transforming efficiency but was itself insufficient to induce transformation. Like the v-fms oncogene product, receptors bearing the activating mutation retained high-affinity binding sites for CSF-1 but were retarded in transport to the cell surface and were phosphorylated on tyrosine in the absence of ligand. Although the activating mutation does not affect the CSF-1 binding site in the receptor extracellular domain, it must induce a conformational change that mimics the effect of ligand binding, resulting in CSF-1-independent signals for cell growth. PMID- 2974322 TI - Inhibition of phosphorolytic degradation of 5-fluoro-2'-deoxyuridine by pyrimidine acyclonucleosides and normal pyrimidine metabolites in rat and beagle tissue homogenates. PMID- 2974323 TI - Inadvertent epidural administration of potassium chloride. A case report. AB - A 24-year-old black female presented for repeat elective Caesarean section. The procedure was performed under epidural anaesthesia. Sufentanil 25 micrograms, intended for postoperative analgesia, was inadvertently diluted to 10 ml with 15 per cent potassium chloride (KCl) instead of preservative-free normal saline (0.9 per cent NaCl). This solution was then injected via an epidural catheter into the epidural space at the conclusion of surgery. Two hours after injection of the sufetanil-KCl mixture, the patient had a level of sensory blockade to T1 and diaphoresis above this level. Painful muscle spasms had also developed below T1. One hour later she developed hypertension which required hydralazine 10 mg and labetalol 25 mg IV for treatment. The patient was treated supportively with oxygen. Dexamethasone 10 mg was administered intravenously to reduce spinal cord oedema. Intravenous diazepam 10 mg and meperidine 75 mg were given for sedation and analgesia. Complete recovery occurred within 12 hours. PMID- 2974324 TI - Vitamin A in acne vulgaris. PMID- 2974325 TI - Tyrosinaemia type II (Richner-Hanhart syndrome)--report of two cases treated with etretinate. PMID- 2974326 TI - Acne fulminans with circulating immune complexes and leukaemoid reaction treated with steroids and azathioprine. PMID- 2974327 TI - Cloning and expression of the atrial natriuretic factor gene. AB - Atrial natriuretic factor (ANF) is a 28-amino acid peptide hormone with potent natriuretic, diuretic and vasodilator properties. Isolation and DNA sequence analysis of rat and human cDNA clones revealed that ANF is synthesized from a 126 amino acid precursor which is highly conserved in both species. Southern blot analysis indicated that the ANF gene is present in a single copy per haploid genome. Both human and rat ANF genes were isolated and showed a similar structural organization which consisted of three exons and two introns. The ANF gene was localized to the short arm of human chromosome 1 and mouse chromosome 4. While atria are the major site of expression of the ANF gene in adult heart, other tissues like ventricles, lung, anterior pituitary, hypothalamus and adrenal synthesize ANF albeit to a much lower extent. In ventricles, ANF mRNA levels are 150 times lower than in atria. However, in cardiac hypertrophy or in congestive heart failure, ventricular ANF mRNA and peptide levels are dramatically (100 fold) increased both in animal models and in humans. This suggests that ventricles are a major site of ANF gene expression in certain pathophysiological conditions and that ANF is not an exclusively atrial peptide as was originally thought. PMID- 2974328 TI - [Possible biomedical uses of alkyl-cyanoacrylates]. PMID- 2974329 TI - [Asymptomatic urinary hospital infections: choice of treatment with drinking water]. PMID- 2974330 TI - [Cotrimazine in the treatment of urinary tract infections in infants]. PMID- 2974331 TI - [Toxic rhabdomyolysis and tubular necrosis in hemlock poisoning. 4 case reports]. PMID- 2974333 TI - [Indications for emergency cesarean section]. PMID- 2974332 TI - [Comparison of the effects of oxprenolol and nifedipine on blood pressure at rest and during physical exercise in essential hypertension]. PMID- 2974334 TI - [Health education and kinesitherapy in chronic low back pain]. PMID- 2974336 TI - [Diagnostic imaging in the evaluation of abdominal masses in childhood]. PMID- 2974335 TI - [A case of lead poisoning in a drug addict. Observations on etiopathogenetic peculiarities]. PMID- 2974337 TI - [Diagnostic imaging in the evaluation of adrenal masses]. PMID- 2974338 TI - [Activation of cellular N-myc oncogene in neuroblastoma]. PMID- 2974339 TI - [Genetic and cytogenetic characteristics of human neuroblastoma]. PMID- 2974340 TI - [Use of calcium heparin in the prevention of thrombosis after arterial revascularization surgery]. PMID- 2974341 TI - [Therapeutic monitoring of hypertensive syndrome in pregnancy]. PMID- 2974342 TI - [Tolerability of nimesulide in the aged]. PMID- 2974343 TI - [Clinical experience with optimized therapy of insulin dependent diabetes mellitus using NovoPen]. PMID- 2974344 TI - [Acute mesenteric infarction. Recent advances in relation to physiopathology, clinical features and therapy]. PMID- 2974345 TI - [Evaluation of the neutralizing activity on gastric acidity of a carbenoxolone antacid formulation using computerized gastric pH-metry in healthy volunteers]. PMID- 2974346 TI - [Ciprofloxacin in the treatment of chronic urinary tract infections]. PMID- 2974347 TI - [Possible teratogenic effects of drugs administered to pregnant women]. PMID- 2974349 TI - [Use of miocamycin in the therapy of cervical infections caused by Chlamydia trachomatis]. PMID- 2974350 TI - [Iatrogenic arrhythmia]. PMID- 2974348 TI - [Evaluation of apoproteins A and B, lipoproteins and plasma lipids during treatment with bezafibrate in various forms of hyperlipoproteinemia]. PMID- 2974351 TI - [Organochlorine pesticides]. PMID- 2974352 TI - [Electrocardiographic changes in kidney transplant under immunodepressive therapy with Cy-A]. PMID- 2974354 TI - [Clinico-bacteriological efficacy, pharmacokinetics and tolerability of netilmicin administered in a daily single dose in bronchopulmonary infections]. PMID- 2974353 TI - [Acute hepatic steatosis. Description of a clinical case of sodium valproate induced acute hepatic steatosis]. PMID- 2974355 TI - [Correction of dietary lactose intolerance by administration of liquid lactase]. PMID- 2974356 TI - [Cardiac changes in dermatomyositis]. PMID- 2974357 TI - [Effects on lipid metabolism of inhibitors of cholesterol synthesis]. PMID- 2974358 TI - [The non-hospital therapeutic approach to patients with acute thoracic pain]. PMID- 2974359 TI - [Focal nodular hyperplasia of the liver]. PMID- 2974360 TI - [Problems of vaccinations during pregnancy]. PMID- 2974361 TI - [The problem of ventricular tachycardia with torsade de pointes and its rational therapy]. PMID- 2974362 TI - [Therapy of hypercholesterolemia with particular reference to inhibitors of cholesterol synthesis]. PMID- 2974363 TI - [Treatment with polyunsaturated phosphatidylcholine of dyslipidemia in pregnancy]. PMID- 2974364 TI - [False values of blood digoxin in patients with acute and chronic liver disease. Role of canrenoate potassium and comparison of radioimmunologic and immunoenzymatic methods]. PMID- 2974365 TI - [Effects of prolonged administration of flunarizine in rats]. PMID- 2974367 TI - [Dissecting aneurysm of the aorta. Role of the internist and possibility of prevention]. PMID- 2974368 TI - [New antibiotic drugs]. PMID- 2974366 TI - [Importance of essential macronutrients in the human organism: the role of chromium]. PMID- 2974369 TI - [Problems of antiparasitic therapy in pregnancy]. PMID- 2974370 TI - [Drug therapy of obesity: current status and future prospectives]. PMID- 2974372 TI - [Use of cefotetan in antibiotic prevention in gynecologic surgical interventions]. PMID- 2974371 TI - [Evaluation of the efficacy and tolerance of enprostil, synthetic analog of prostaglandin E2, in a clinical study in double-blind vs. placebo]. PMID- 2974373 TI - [Therapy of herpes simplex virus infections with acyclovir cream]. PMID- 2974374 TI - [Nosocomial infections]. PMID- 2974376 TI - [Superficial and deep EEG recordings of valerian-related drugs]. PMID- 2974377 TI - [Ketoprofen lysine treatment of painful manifestations of spinal osteoarthrosis]. PMID- 2974375 TI - [Problems of mineral therapy in pregnancy]. PMID- 2974378 TI - [Endoscopic and clinical evaluation of the therapeutic action of a bicarbonate calcium mineral water in patients with gastroduodenitis]. PMID- 2974380 TI - [Urinary incontinence caused by unstable bladder]. PMID- 2974379 TI - [Acute respiratory insufficiency. Etiological, physiopathological clinical and therapeutic aspects]. PMID- 2974381 TI - [Venous diseases of the lower limbs and pulmonary embolism]. PMID- 2974382 TI - [Efficacy and tolerability of cefoperazone in patients with nosocomial infections]. PMID- 2974383 TI - [Neuroleptic malignant syndrome. Clinical and therapeutic considerations]. PMID- 2974386 TI - [Comparison of steady state pharmacokinetics of isosorbide-5-mononitrate in microgranular formulation or hydrophilic matrix]. PMID- 2974384 TI - [Proglumetacin in the treatment of rheumatoid arthritis and arthrosis]. PMID- 2974385 TI - [Current diagnostic-therapeutic protocols in upper digestive hemorrhage]. PMID- 2974387 TI - [Bacterial agents responsible for urinary tract infections. III. A clinico statistical contribution]. PMID- 2974388 TI - [Traumatic pathology of the abdomen: the role of CT]. PMID- 2974389 TI - [The number of cases of AIDS in the USSR have surpassed 30]. PMID- 2974390 TI - [Women die of AIDS earlier than men]. PMID- 2974391 TI - [Use of beta interferon in ophthalmology. Description of a series of cases of viral diseases of the anterior segment]. PMID- 2974392 TI - [Experience with phosphatidylserine treatment of patients with cognitive and behavioral decline]. PMID- 2974393 TI - [Leukemoid reaction in acute alcoholic hepatitis]. PMID- 2974394 TI - [Role of trans-parieto-hepatic cholangiography in cholestatic jaundice]. PMID- 2974395 TI - [Use of cefaclor monohydrate in infections of the upper and lower respiratory tract]. PMID- 2974396 TI - [Heparin prophylaxis of deep venous thrombosis and pulmonary embolism in medical patients]. PMID- 2974397 TI - [Luciani-Wenckeback period and clonidine. Presentation of a case]. PMID- 2974398 TI - [Computerized tomography in the evaluation of serous effusions]. PMID- 2974399 TI - [Disinfectants: availability and mechanisms of action]. PMID- 2974400 TI - [Surgical therapy of ametropia]. PMID- 2974401 TI - [Sodium diclofenac in the treatment of biliary pain: a controlled study using hyoscine N-butylbromide]. PMID- 2974402 TI - [Comparison of an antibiotic combination (amoxicillin + clavulanic acid) and cefaclor in the treatment of acute or flare-ups of chronic bronchopneumopathies]. PMID- 2974403 TI - [1% tioconazole powder in the treatment of dermatomycoses]. PMID- 2974404 TI - [Rational and irrational approach to the inotropic agent therapy of heart failure]. PMID- 2974405 TI - [Long-time treatment with full-dosage ranitidine in a case of myelodyserythropoiesis (blastic leukosis) treated with prednisone and antiblastic agents]. PMID- 2974406 TI - [Nodular lymphoid hyperplasia of the colon]. PMID- 2974407 TI - [Pesticides: organophosphorus compounds]. PMID- 2974408 TI - Hyaluronic acid and other glycosaminoglycans of the human palatal tonsils. AB - The aim of this study was to determine whether the proportions of individual glycosaminoglycans (GAGs) change with bacterial infection of the human palatal tonsils and after recovery as compared with the physiological state of the palatal tonsils. In tonsillitis the greatest changes were related to hyaluronic acid (decrease) and heparin (increase). In the cured palatal tonsils the content of GAG components was found to return to that of the physiological state of the tonsils. The changes in GAG components in the cured palatal tonsils may possible contribute to the reversal of the human tonsillitis. The supposition is drawn that also physiological function of the cured tonsils is resumed. PMID- 2974410 TI - The subcorneal pustular drug eruption: an example induced by norfloxacin. AB - In the patient described here, norfloxacin induced a generalized erythematous subcorneal pustular drug eruption associated with photosensitivity and fever, proven by challenge. Any pustular eruption with fever should engender a careful search for a drug that may have caused this reaction. PMID- 2974409 TI - Dermatologic manifestations in users of video display terminals. PMID- 2974411 TI - Propylene glycol in the treatment of seborrheic dermatitis of the scalp: a double blind study. AB - Thirty-nine patients with seborrheic dermatitis of the scalp were treated in a double-blind controlled study with a solution containing either 15 percent propylene glycol, 50 percent ethanol, and 35 percent water, or a solution containing 50 percent ethanol and 50 percent water. Two patients did not return for follow-up. Sixteen of eighteen (89 percent) in the group treated with propylene glycol showed healing, compared to six of nineteen (32 percent) in the other group. In twenty patients quantitative cultures for Pityrosporum orbiculare were taken. The number of organisms was reduced significantly after treatment with the propylene-glycol-containing solution but not in the other group. This propylene-glycol-containing solution was easy to apply, cosmetically attractive, and may be an alternative to corticosteroids for the treatment of seborrheic dermatitis of the scalp. PMID- 2974412 TI - Powerlessness in cervical spinal cord injury patients. PMID- 2974413 TI - Choosing and using hepatitis B vaccine. PMID- 2974414 TI - [Modification of atrial natriuretic factor by the AV transmission time in pacemaker patients]. AB - The possible relationship of variations in the programmed a-v interval on the peripheral-venous plasma level of the atrial natriuretic factor (ANF) was investigated in nine patients with dual-chamber pacemakers (DDD). During atrial and ventricular stimulation at 82 beats/min the pacemakers of the resting patients were programmed to give successive a-v intervals of 200, 175, 150, 125, 100, 50 and 15 ms. Blood samples were obtained from a peripheral arm vein ten minutes after each change of programme. ANF was extracted from each sample and determined by radioimmunoassay. Both the absolutely and relatively lowest plasma ANF level was obtained at an a-v interval of 175 ms. Lengthening the interval to 200 ms or shortening it to 150 ms or less increased the level. It is possible that both "too long" and "too short" a-v intervals produce an increase in atrial pressure and stretch, to which ANF reacts highly sensitively with an increase in peripheral-venous plasma levels of ANF. PMID- 2974415 TI - Influence of a diet rich in fish oil on blood pressure, body weight and cardiac hypertrophy in spontaneously hypertensive rats. AB - The effects of a diet rich in fish oil on arterial blood pressure, body weight, left ventricular weight and heart rate have been investigated in 8 month old spontaneously hypertensive male rats (SHR) as compared to age-matched hypertensive controls. A diet containing 10% fish oil decreased blood pressure by about 40 mmHg within 20 days of starting the experiment, and this effect persisted over the observation period of 80 days. Permitting the animals free access to food, the body weight of the diet group increased by 25%. The degree of hypertrophy as evaluated by relating left ventricular weight to tibial length was significantly reduced (10%) in the diet fed group. Heart rate was increased by 53%. The study demonstrates that a diet rich in fish oil can lower arterial blood pressure over several weeks without a recognizable loss in function despite a considerable increase in body weight. It can be assumed that a more marked regression of left ventricular hypertrophy is counteracted by a reflex increase in sympathetic efferentation to the heart. PMID- 2974416 TI - Clinical and neurobiological findings in children suffering from tic disease following treatment with tiapride. AB - Tiapride, a substituted benzamide derivative, possesses good clinical antidyskinetic properties due to its DA2-blocking activities. It has been shown to be clinically effective in the treatment of tic disease in children. In order to study tiapride's antidyskinetic properties in the treatment of the tic syndrome in children, we conducted a simple, placebo-controlled study on 10 children followed by a double-blind crossover study on 17 children. Tiapride was shown to have a positive therapeutic effect on tics in children; whereas it has no adverse effects on neuropsychologically measurable cognitive performances in children. Neurophysiological parameters such as the EEG frequency analysis and sensory evoked potentials were not affected by tiapride, nor was the neurosecretory, hypothalamic-hypophyseal regulation of the sex hormones, thyroid stimulating hormone, growth hormone, or thyroid hormone impaired. The hyperprolactinemia caused by tiapride's dopaminergic properties was moderate and restricted to the duration of therapy. PMID- 2974417 TI - Possible site of bronchodilation due to inhaled procaterol aerosol in asthmatic patients. AB - We studied the effective site of an inhaled aerosol of procaterol, a beta 2 selective adrenergic bronchodilator, in 8 asthmatic patients whose basal lung functions are almost within the normal range in both slow vital capacity (VC) and forced expiratory volume in one second (FEV1.0), and are free from asthmatic attack. In patients who had received procaterol 30 min after inhalation of aerosol, there was no significant change in VC, although FEV1.0, maximal expiratory flow at 50% VC (V50), maximal expiratory flow at 25% VC (V25) and maximal expiratory flow at 30% VC of partial maximal expiratory flow volume curve (V30p) improved significantly. On the other hand, in those who had received placebo, none of the parameters changed. Furthermore, R1 decreased and C0.5 increased significantly during the first 5 min after inhalation of procaterol aerosol. After an interval of 5 min, R1 did not change any further, while C0.5 continued to improve until 30 min after inhalation of procaterol. These results suggest that procaterol may first dilate the large airway and then may gradually dilate the small airway in bronchial asthma. PMID- 2974418 TI - The pharmacokinetics of doxifluridine and 5-fluorouracil after single intravenous infusions of doxifluridine to patients with colorectal cancer. AB - The disposition kinetics of a new 5-fluorouracil prodrug, 5'-deoxy-5 fluorouridine (5'dFUR, doxifluridine), were investigated in six patients with colorectal carcinoma. Each patient randomly received two single intravenous doses of 5'dFUR (2 and 4 g.m-2) on separate days. Plasma concentrations of 5'dFUR fell rapidly with terminal half-lives ranging from 16.1 to 27.7 min. A disproportionate increase in the area under the curve with increasing dose was seen in most patients. Doubling the dose resulted in a 40% decrease in nonrenal clearance (0.60 to 0.37 l.min-1) but no apparent change in renal clearance (0.32 to 0.29 l.min-1) or steady-state apparent volume of distribution (19.8 to 20.4 l). The mechanism for dose-dependence of 5'dFUR appears to be primarily due to nonlinear elimination associated with nonrenal processes rather than nonlinear plasma protein or tissue binding. PMID- 2974419 TI - The in vitro stability of doxifluridine in whole blood and plasma. PMID- 2974420 TI - Limiting dilution analysis of proliferative responses in human lymphocyte populations defined by the monoclonal antibody UCHL1: implications for differential CD45 expression in T cell memory formation. AB - Using limiting dilution analysis, we investigated proliferative responses of UCHL1+ and UCHL1- T cell populations to compare the precursor cell frequencies following recall and alloantigen stimulation, and the complexity of cellular interactions within UCHL1+ and UCHL1- populations. We find high frequencies of recall antigen responses among UCHL1+, but not UCHL1- T cells. In contrast, both populations contain similar frequencies of alloantigen responsive cells. Our results are consistent with single-hit kinetics in recall as well as alloantigen responses and show no complex cellular interactions within the responding populations. In conclusion, the difference between the recall antigen response of UCHL1+ and UCHL1- cells observed in conventional proliferation assays is most probably due to a high frequency of recall antigen-responsive UCHL1+ cells and not to suppressive phenomena in the UCHL1- population. These data suggest that memory T cells are largely of the UCHL1+ phenotype. The relation of post-thymic T cell maturation and differential CD45 expression is briefly discussed. PMID- 2974421 TI - Cloned T helper cells reverting to a resting state develop increasing sensitivity in their antigen-mediated interaction with accessory cells. AB - A cloned murine T cell line, KIII5, specific for the polypeptide poly-L(Tyr,Glu) poly-D,L-Ala--poly-L-Lys [(T,G)-A--L] was compared at different stages after antigenic stimulation with respect to the conditions required for the reinduction of growth by varying concentrations of antigen presented on different types of accessory cells (AC). We show that the dose of antigen necessary for inducing half maximal proliferation in the presence of splenic AC shifts to considerably lower concentrations when the T cell blasts revert to a resting state (100 micrograms/ml on day 7 to 10 micrograms/ml on day 21-35). During the same time period the expression of interleukin 2 (IL2) receptor and the reactivity to IL2 decline. However, no direct correlation between the increasing sensitivity to antigen and the decreasing reactivity to IL2 appears to exist. With peritoneal AC "early" T cells (day 7) did not respond to (T,G)-A--L at all, but in the course of "aging" responsiveness increased and finally reached the same level as in the presence of splenic AC, although at a higher antigen dose (100 micrograms/ml on day 35-45). Furthermore, the antigen-induced proliferation of "aging" T cells became more resistant to inhibition both by anti-L3T4 and anti-T cell receptor antibodies. Two alternative interpretations of these data are possible: antigen activated T cells, while gradually reverting to a resting state, interact more avidly with antigen-presenting cells or the triggering threshold of the T cells is decreasing. PMID- 2974422 TI - Functional interaction between B cell subpopulations defined by CD23 expression. AB - Using the CD23 monoclonal antibody (mAb) MHM6 and sheep anti-mouse Ig bound to magnetic beads we have obtained highly purified populations of MHM6+ and MHM6- tonsil B cells. We have found that the increased expression of MHM6 reactivity seen on B cells after activation results from up-regulation of antigen on cells already weakly positive and not from expression of new antigen on the previously negative population. The strong proliferative responses of MHM6+ cells seen in the presence of anti-IgM (alpha mu) and interleukin 4 (IL4) or the CDw40 mAb G28 5, and with Staphylococcus aureus Cowan I (SAC), and to a lesser extent with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), resemble that seen among unfractionated B cells. In contrast, the MHM6- population cultured alone responds only weakly to alpha mu + G28-5 or SAC and exhibits virtually no response to alpha mu + IL4 or TPA. With all these mitogenic stimuli, tritiated thymidine uptake by the MHM6- population is augmented three- to sixfold by the addition of mitomycin C (MC)-treated MHM6+ cells. Pretreatment of cells with anti leukocyte functional antigen 1 mAb has little effect on the subsequent proliferation of the MHM6- population but shows cell contact to be critical for the proliferation of MHM6+ cells. Such pretreatment has revealed that the functional interaction observed between MHM6+ and MHM6- cells is dependent on both cell contact and the presence of an MHM6+ cell-derived soluble component. We have found that addition of soluble CD23, purified from Epstein-Barr virus transformed lymphoblastoid cell line supernatant, increases the proliferative response of MHM6- tonsil B cells to mitogenic stimuli in the presence of inactivated MHM6+ cells but has no effect on proliferation when MHM6+ cells are absent. By way of contrast to normal B lymphocytes, we have examined functional responses of prolymphocytic leukemia (PLL) B cells. Although these cells, when freshly isolated, show comparable levels of CD23 expression to normal B cells, this expression is not increased upon activation. In addition, in contrast to normal B cells, the PLL MHM6- population cultured alone shows a strong proliferative response to various mitogenic stimuli, comparable to that of MHM6+ or unfractionated cells, and this response is not augmented by the addition of MC treated MHM6+ cells. Thus, a novel functional interaction is described between normal, but not leukemic, B cell populations defined by their expression of CD23.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2974423 TI - Autoreactive T cells in mercury-induced autoimmunity. Demonstration by limiting dilution analysis. AB - Mercuric chloride is responsible in Brown-Norway rats for an autoimmune disease that is autoregulated. Previous studies have shown that this agent induces T cell dependent polyclonal B cell activation in these rats. Evidence has also been obtained for the existence of autoreactive T cells which play a role in the evolution of this process. In the present study, limiting dilution analysis was used to demonstrate that (a) frequent autoreactive T helper cells which proliferate in the presence of T cells from HgCl2-injected rats are present from day 4; (b) frequent auto-anti-Ia T helper cells which recognize normal B cells as well as B cells from HgCl2-injected rats appear from day 6; and (c) less frequent T suppressor cells which could play a role in autoregulation emerge from day 14. PMID- 2974424 TI - Establishment of tumor-specific immunotherapy model utilizing vaccinia virus reactive helper T cell activity. AB - It was previously demonstrated that preimmunization of mice with live vaccinia virus (VV) and subsequent immunization with VV-infected (modified), syngeneic tumor cells resulted in enhanced induction of tumor-specific immunity through a cellular collaboration between VV-reactive helper T (VV-Th) cells and tumor specific effector cell precursors. On the basis of this augmenting system, a tumor-specific immunotherapy model was established in which a growing tumor regressed. C3H/HeN mice were pretreated with cyclophosphamide to eliminate nonspecific suppressor T cell activity and subsequently inoculated (primed) s.c. with live VV, leading to augmented induction of VV-Th cell activities. Four weeks later, the mice were inoculated i.d. with syngeneic X5563 myeloma cells. Six days after the tumor cell inoculation, live VV was injected into the tumor mass three times at 2-day intervals. Seven of ten mice which had received VV priming and subsequent VV injection into the tumor mass exhibited complete tumor regression. On the contrary, mice which had received mere intratumoral VV injection in the absence of VV priming failed to exhibit appreciable tumor regression. Mice whose tumor had completely regressed (regressor mice) by the above VV immunotherapy were shown to have acquired systemic anti-tumor immunity, which was confirmed by a challenge with syngeneic tumor cells after the tumor regression. In vitro analysis of these immune mice revealed that potent tumor-specific cytotoxic T lymphocyte responses were preferentially induced, but with no detectable anti tumor antibody responses. Such a potent tumor-specific immunity was not observed in mice which had received an intratumoral VV injection in the absence of VV priming. Thus the results clearly indicate that the tumor regression was accompanied by concurrent generation of a potent tumor-specific immunity, suggesting that the cellular collaboration between VV-Th cells and tumor-specific effector cell precursors is also functioning in this VV-immunotherapy protocol, likewise the immunoprophylactic model. Therefore, the present model provides an effective maneuver for tumor-specific immunotherapy and this system will be theoretically applicable to human cancer treatment. PMID- 2974425 TI - Age-associated increase in the expression of T cell antigen receptor gamma chain genes in mice. AB - T cell function generally declines with age. To determine the underlying cause of the age-related decline, we compared the expression levels of T cell antigen receptor genes encoding gamma, beta and alpha chains in the lymphoid tissues of young (8 wk old) vs. aged (40 wk old) mice. An age-associated increase in gamma chain gene transcripts was evident in the thymocytes, spleen cells and mesenteric lymph node cells. Aged mice had a relatively high proportion of CD3+CD4-CD8- cells but a reduced level of CD3+CD4-CD8+ cells in the lymphoid tissues, as compared with young counterparts. The allo-reactivity in the lymphoid cells, as assessed by mixed lymphocyte reaction (MLR) responses, decreased with advancing age. On the contrary, increased levels of syngeneic MLR and spontaneous cytolytic activity were noted in the lymphoid cells of aged mice, as compared with findings in their young counterparts. A remarkable increase in the number of CD3+CD4-CD8- cells and level of the gamma chain gene messages was also detected in the responder cells of day 4 syngeneic MLR culture from aged mice. An increase in the number of T cells bearing gamma chain may be related to the alterations in immunological functions in aged mice. PMID- 2974426 TI - Immune responses to an adjuvant-free native syngeneic myeloma protein (M315). AB - Myeloma protein 315 (M315; isotype IgA, lambda 2) is used in this report as a model to explore the immunogenicity of a syngeneic Ig under nearly physiological conditions. We have previously shown that a synthetic peptide spanning the mutated HV3 loop of the L-315 chain, when emulsified in complete Freund's adjuvant, elicits T helper cells (Th) that respond to a boost with L-315 or M315, indicating that M315 is recognized as a processed protein antigen. We now show that the adjuvant-free 7S monomer of native or of mildly reduced and alkylated M315, given in divided doses totalling 300 or 800 micrograms to BALB/c mice, induced persistent anti-M315 antibodies (Ab), a large part of which was IgG1 directed mainly to idiotypes (Id) associated with M315's hapten-binding site. Polymers of M315 IgA (800 micrograms) failed to induce Ab, due probably to their rapid clearance into bile. Short-term treatment with anti-CD4 monoclonal Ab GK1.5 at the time of priming with 7S M315 inhibited the responses almost completely. The spleens of M315-immune mice contained Th that recognized the L-chain subunit of M315 as a carrier indicating that these Th did not require an assembled (VH VL) pair of 315 V regions to be activated. We also observed low amounts of Ab specific for epitopes of the C alpha region. This evidence opens the possibility that a distinct autoimmune pathway exists for elicitation of rheumatoid factor (RF; autoAb to Fc gamma) that involves help to RF-producing B cells by Id specific Th. We suggest that these Th recognize V-region peptides from IgG that have been captured, processed and presented by these B cells. PMID- 2974427 TI - Clonal analysis of CD4-CD8- human thymocytes expressing a T cell receptor gamma/delta chain. Direct evidence for the de novo expression of CD8 surface antigen and of cytolytic activity against tumor targets. AB - CD4-CD8- human thymocytes were obtained by treating total thymocyte suspensions with anti-CD4 and anti-CD8 monoclonal antibodies (mAb) and complement. The resulting cell populations contained virtually no CD4+, CD8+ or WT31+ cells and 17-65% CD3+ cells. In addition, analysis of cell reactivity with delta-TCS-1 mAb (specific for the C gamma 2-encoded, nondisulfide-linked form of TcR gamma/delta), revealed the presence of a variable proportion of delta-TCS-1+ cells (the % of delta-TCS-1+ cells were lower than the percentage of CD3+ cells). Upon culture in recombinant interleukin 2 (IL2, in the presence of irradiated mononuclear cells), CD4-CD8- thymocytes underwent extensive proliferation. In addition, a progressive increase of CD8+ cells (but not of CD4+ or WT31+ cells) could be detected. Cells also progressively acquired cytolytic activity against K 562 or fresh melanoma cells. Fresh CD4-CD8- thymocytes were cloned under limiting dilution conditions. The cloning efficiencies were relatively high (1/3 cells); in addition, virtually all the clonal progenies obtained displayed cytolytic activity and expressed the CD3+WT31-delta-TCS-1+ surface phenotype. About half of the clones analyzed were CD8+, whereas none expressed CD4 antigens. We conclude that (a) only delta-TCS-1-reactive, TcR gamma/delta+ cells can be isolated from CD4-CD8- thymocytes cultured in IL2, and (b) the expression of CD8 antigen and of cytolytic activity reflects a true in vitro phenotypic change of CD8-, noncytolytic precursors (and not the preferential growth of few contaminating cells). PMID- 2974428 TI - Danazol and medroxyprogesterone acetate inefficacious in the treatment of infertility in endometriosis. AB - Danazol (200 mg three times a day) and medroxyprogesterone acetate (MPA, 100 mg a day) were compared with placebo in the treatment of infertility of patients with endometriosis. Twenty-seven patients had medical therapy alone for 6 months, and 22 patients received it after conservative surgery. The clinical characteristics of the patients in the danazol group (n = 18), the MPA group (n = 17), and the placebo group (n = 14) were comparable to each other. The follow-up time was 30 months. The cumulative pregnancy rates, 33% in the danazol group (n = 6), 42% in the MPA group (n = 7), and 46% in the placebo group (n = 6), did not differ significantly from each other. The time to pregnancy after the start of therapy was 17.7 +/- 8.4 (standard deviation [SD]) months in the danazol group, 18.0 +/- 9.0 months in the MPA group and 10.0 +/- 5.8 months in the placebo group with no significant difference between the groups. The abortion rate was 26%, and there was no significant difference among the groups. Cox multivariant analysis did reveal ovarian endometriosis a prognostically significant negative indicator as regards fecundation in endometriosis (P less than 0.05). In summary, correction of infertility alone does not appear to be an indication for the use of danazol or MPA in the treatment of endometriosis, and ovarian endometriotic lesions but not peritoneal ones do make a worse prognosis as regards fecundation in endometriosis. PMID- 2974429 TI - Ovarian hyperstimulation syndrome following D-Trp-6 luteinizing hormone-releasing hormone microcapsules and menotropin for in vitro fertilization. AB - In 143 cycles of in vitro fertilization the ovarian hyperstimulation syndrome (OHSS) occurred in 12 (8.4%) cycles. Six were in the moderate form and 6 severe. Ovarian stimulation by menotropins was preceded by induction of hypopituitary hypogonadism using D-Trp6-LH-RH microcapsules. The OHSS cycles are characterized by improved ovarian response expressed by the increased serum levels of estradiol, number of follicles, oocytes, embryos and pregnancy rate as compared to cycles with no OHSS. All patients recovered uneventfully. The follicular puncture did not have the suggested protective effect against OHSS. It is suggested that the substantial incidence of OHSS is probably related to the excessive ovarian stimulation not interrupted by early luteinization which is practically abolished by this protocol. The role of the given luteal hCG doses in the genesis of OHSS is questioned. PMID- 2974430 TI - Gamete intrafallopian tube transfer (GIFT): making laparoscopy more than "diagnostic". AB - Diagnostic laparoscopy is commonly performed on patients as part of a complete infertility investigation. Recently published protocols have investigated the efficacy of using empiric ovulation induction, intrauterine insemination, or both before beginning in vitro fertilization. Because many patients enrolled in these protocols will be exposed to both ovulation induction and diagnostic laparoscopy, the authors reasoned that it would be more effective to begin ovulation induction at the time of the proposed diagnostic laparoscopy in order to allow gamete intrafallopian tube transfer (GIFT) at the same time. Twenty-five nulliparous patients underwent diagnostic GIFT. There were eight continuing pregnancies in this group (32%). Diagnostic GIFT is a more effective use of the opportunity provided by laparoscopy than diagnostic laparoscopy alone. PMID- 2974431 TI - Ultrasound-guided fallopian tube catheterization per vaginum: a feasibility study with the use of laparoscopic control. AB - The fallopian tube can be cannulated per vaginum under ultrasound control. The sensation and ultrasound appearance of smooth passage without visible kinking are accurate predictors of success. The average time taken for each cannulation was 7.2 minutes. Adequate practice in non treatment cycles is essential before proceeding to cell transfer, while further modification of the catheters used may improve overall success. PMID- 2974432 TI - Child health nurses--caring for our nation's youth. Safety seats play major role for safe transportation. PMID- 2974433 TI - The role of prostaglandin E in the natural suppressor activity of spleen cells from newborn rats. AB - Spleen cells from newborn to 2-day-old, but not 3-day-old or older, rats suppress the activity of cells from adult animals in xenogeneic local graft-versus-host (GVH) assay. The duration of neonatal suppressor cell activity can be significantly prolonged by treating newborn rats with exogenous prostaglandin E2 (PGE2), and the suppression was completely abolished by administration of PGE inhibitor indomethacin. The results thus demonstrate an involvement of PGE2 in the mechanism of action of neonatal suppressor cells and may explain some discrepancies concerning the nature of natural suppressor cells present in newborn animals. PMID- 2974434 TI - [Medico-genetic study of the population of Kostroma Province. VII. Approaches to the study of the hereditary pathology load in the population of Kostroma]. AB - Complex medical-genetic study was performed in the center of Kostroma Province with the population of about 250 thousands. A method for ascertainment of patients was developed and the information value of different sources of a registration system available is given. The complete algorithm of calculation of both the segregation frequency and the fraction of sporadic cases for autosomal recessive diseases is presented. PMID- 2974435 TI - [Hereditary pathology of the nervous system in the urban population of the Kazakh SSR]. AB - The data are presented on clinical forms and prevalence of neurological diseases in two regions of Kazakhstan, where about 2180600 inhabitants live. The data may constitute a basis for regional registration of neurological diseases and planning extensive specific genetic counselling concerning the diseases in the regions under study. PMID- 2974436 TI - Presence of abnormal synaptonemal complexes in heterothallic species of Neurospora. AB - Synaptonemal complex abnormalities are frequent in reconstructed meiotic prophase nuclei of Neurospora crassa and Neurospora intermedia. Three kinds of synaptonemal complex anomalies were seen: lateral component splits, lateral component junctions, and multiple complexes. The anomalies apparently are formed during or after the pairing process, as they were not seen in the largely unpaired early zygotene chromosomes. Their presence at all the other substages from mid-zygotene to late pachytene indicates that they are not eliminated before the synaptonemal complex decomposes at diplotene. Abnormal synaptonemal complexes were seen in all 19 crosses of N. crassa and N. intermedia that were examined, including matings between standard laboratory strains, inversions, Spore killers, and strains collected from nature. The frequency of affected nuclei and degree of abnormality within a nucleus varied in different matings. No abnormalities were present in the homothallic species Neurospora africana and Neurospora terricola. Structural chromosome aberrations, introgression, and heterozygosity have been eliminated as causes for pairing disorder. The abnormal synaptonemal complexes seemingly do not interfere with normal ascus development and ascospore formation. The affected nuclei are not aborted during meiotic prophase, nor are they eliminated by abortion of mature asci. The abnormal meiocytes do not lead to aneuploidy, as judged by the low frequency of white ascospores in crosses between wild type strains that have many abnormalities. Thus, the abnormal synatonemal complexes do not appear to prevent chiasma formation between homologues. PMID- 2974437 TI - [Substantiation of time-differentiated maximum permissible levels of sevin in atmospheric air]. PMID- 2974438 TI - [Experience of the scientific research institute in providing practical assistance in public health]. PMID- 2974439 TI - [Hygienic problems with regard to architectural and planning decisions and the design and construction of rural schools in Byelorussia]. PMID- 2974440 TI - [Activities of the People's "Health" University for students of special secondary schools]. PMID- 2974441 TI - [Physical development of workers in various categories of engineering industry]. PMID- 2974442 TI - Use of decision analysis in a complicated case of renovascular hypertension. PMID- 2974443 TI - The international standard for atrial natriuretic factor. Calibration by an international collaborative study. AB - An ampouled preparation of human atrial natriuretic factor, ANF-(99-126), was evaluated by 23 laboratories in 10 countries for its suitability to serve as the international standard for ANF. The preparation was calibrated by radioimmunoassay, radioreceptor binding assay, and bioassay and was shown to have satisfactory stability and biological activity. Estimates of the ANF content of a set of specimens of plasma in terms of the standard showed agreement in ranking order when the ANF was extracted prior to assay. However, estimates of the ANF content of the plasmas in terms of either the international standard or the various local standards varied widely among laboratories. On the basis of the results reported here, with the agreement of the participants in the study and with the authorization of the Expert Committee on Biological Standardization of the World Health Organization, the preparation coded 85/669 was established in 1987 as the international standard for ANF, with a defined potency of 2.5 international units per ampoule. PMID- 2974444 TI - T cell immunophenotypes and DR antigen expression in intravenous drug users. Relationship to human immunodeficiency virus serology. AB - Thirty-six intravenous drug users were studied for peripheral blood mononuclear cell (PBMC) immunophenotypes and human immunodeficiency virus (HIV) serological profiles. This population has a high risk for developing HIV infection. Half (18/36) were HIV antibody (Ab) negative (-) and half were positive (+). Total T lymphocytes (CD3+ and CD2+) were not different between HIV Ab-negative and HIV positive groups. Unactivated T(CD3+DR-) cells/mm3 were less (p = 0.003) in HIV Ab positive patients (1,467 +/- 628) compared to HIV Ab-negative patients (2,190 +/- 695). T-helper (CD4+) cells/mm3 were also less in HIV Ab-positive patients (762 +/- 344 vs. 1,161 +/- 419, p = 0.005). The most significant difference was in activated T lymphocyte CD3+DR+) percentages where the mean was 9.6% in those HIV Ab-positive compared to 3.8% in seronegatives (p less than 0.001). Preliminary studies showed that in vitro naloxone treatment of PBMC had no effects on immunophenotypic expression except for CD3+DR+ lymphocytes, where a significant reduction was observed in the HIV Ab-positive group (p = 0.022) but not in the HIV ab-negative group. These findings suggest that in certain populations, activated T cells may be an early manifestation of HIV infection. PMID- 2974445 TI - Influence of an Hymenolepis diminuta infection on IgE and IgA bound to mouse intestinal eosinophils. AB - Infection of mice with Hymenolepis diminuta, which is an 'exclusively' intestinal cestode, affects the number of eosinophils and non-eosinophilic cells with IgE or IgA on their surface in the lamina propria. Presence of IgE on eosinophils is basically a primary infection response, while after reinfection the response is primarily characterized by IgA. For IgE- as well as for IgA-bearing eosinophils the response is most abundant in the second quarter of the intestine which is the parasite's preferred habitat. For non-eosinophilic cells the effect is smaller and limited to the IgE-bearing cells, with the most significant effect in the second quarter of the intestine. PMID- 2974446 TI - Does velocity of dilatation influence the result of dilatation? A postmortem study. AB - We evaluated the influence of velocity of dilatation on the success of and the vascular damage produced by dilatation. Nonatheroslerotic segments of distal superficial femoral arteries were dilated "fast" (n = 69) or "slow" (n = 45) under standard conditions with balloon catheters. The arterial wall was overstretched between 1% and more than 60%. The success of dilatation rose continuously with increasing overstretching during dilatation. The damage to the arterial wall by dilatation increased discontinuously with increasing overstretching. Dilatation success and damage patterns did not differ in the "slow" and "fast" groups. PMID- 2974447 TI - Section 504 of the Rehabilitation Act, HIV and AIDS: legal implications. PMID- 2974448 TI - Mechanism of acquired resistance to methotrexate in P388 murine leukemia cells and in their doxorubicin-resistant subline. AB - The mechanisms of acquired resistance to MTX were studied in P388 murine leukemia cell lines that were sensitive or resistant to ADR. The rate of MTX accumulation in ADR-sensitive cells that have acquired resistance to MTX was found to be lower than that measured in cells that were sensitive to both drugs. Furthermore, in contrast to drug-sensitive cells, in the ADR-sensitive MTX-resistant cells, most of the intracellular MTX (86.2%) was bound and MTX polyglutamation was not detected. The initial rate of MTX accumulation in cells that were resistant to both drugs was comparable to that measured in cells that were sensitive to both drugs or that were resistant only to ADR. However, in the cells that were resistant to both drugs, the rate of MTX accumulation was maintained at its initial level for a period that was considerably longer than that found in the other cell lines. After 3 h of exposure to MTX, the accumulation of MTX in cells that were resistant to both drugs was fourfold higher than that measured in cells that were sensitive to both drugs. Furthermore, while 65 to 70% of the intracellular MTX was free, in cells sensitive to both drugs, or resistant only to ADR, the corresponding value in cells that were resistant to both drugs was less than 1.5%, and a much lower proportion of the MTX was polyglutamated. The sensitivity to TMQ of ADR-sensitive, MTX-resistant cells was similar to that found in cells that were sensitive to ADR and MTX. However, ADR-resistant cells, sensitive or resistant to MTX, were markedly resistant to TMQ. The sensitivity of ADR-resistant MTX-sensitive cells to TMQ was restored by the presence of 10 microM verapamil. Such an effect was not observed in cells resistant to both drugs. It is suggested that P388 cells that have previously acquired resistance to ADR, when now selected by MTX, retain the MTX-transport system (in contrast to ADR-sensitive, MTX-resistant cells) and become resistant to MTX by increasing the activity of DHFR. The results obtained in ADR-resistant cells also suggested that resistance to TMQ was part of the multidrug resistance phenomenon. PMID- 2974450 TI - Teicoplanin revisited. PMID- 2974451 TI - Medical students who care. Fighting AIDS Through Education. PMID- 2974449 TI - Effect of PGE1 treatment on in vitro thymocyte function of normal and autoimmune mice. AB - Autoimmune New Zealand (NZ) mice exhibit a broad spectrum of T and B cell disorders. These include abnormally high levels of terminal deoxynucleotidyl transferase-positive immature T cells in bone marrow and thymus. We have shown previously that prostaglandin E1 (PGE1) treatment of the NZB/NZW F1 hybrid, a murine model of systemic lupus erythematosus (SLE), reduces to normal the percentage of immature terminal deoxynucleotidyl transferase-positive cells in bone marrow and thymus, and prevents the immune complex-induced nephritis which kills these animals. We report here that short-term (1-5 days) treatment of NZB/W mice with PGE1 increases thymocyte responsiveness to mitogens and alloantigens. The majority (greater than 90%) of cortical thymocytes agglutinated by peanut lectin (PNA+) are depleted by PGE1 treatment. However, a small population of highly functional cells persists in the PNA+ fraction after PGE1 treatment. PGE1 appears to have little or no effect on the PNA-negative (medullary) fraction of thymocytes. Our data suggest that PGE1 may exert its therapeutic effect in NZ mice by increasing the functional maturity of immature T cells. PMID- 2974452 TI - Newer concepts in the diagnosis and treatment of ARDS. PMID- 2974453 TI - The physician as a community volunteer. PMID- 2974455 TI - Adenocarcinoid of appendix. PMID- 2974454 TI - Reversal of pure red cell aplasia by cyclosporin-A in a patient with chronic lymphocytic leukemia. PMID- 2974456 TI - Stimulatory effects of androgen and antiandrogen on the in vitro proliferation of human mammary carcinoma cells. AB - The proliferation of three mammary carcinoma cell lines was explored for the effectiveness of dihydrotestosterone (DHT) and the antiandrogenic substances cyproterone acetate (CPA) or hydroxyflutamide. The cell growth, determined in multiple experimental cultures of the estrogen-sensitive lines MCF-7 and EFM-19, was stimulated by 10(-9) M to 10(-6) M DHT, whereas estrogen-resistant MFM-21 cells were unresponsive to the hormonal factors applied. Growth-promoting effects of 10(-8) M to 10(-6) M CPA were detected in cultures of those cell lines which were sensitive to estrogen and androgen. Competition experiments with DHT and the antiandrogens suggested involvement of the androgen receptor in the stimulation of cell growth by CPA. Participation of the estrogen receptor was excluded by lack of competition between CPA and the enhancement of proliferation by estradiol 17 beta. At the receptor level the antiandrogens were able to compete with androgen binding. The results of the study demonstrate androgenic properties of CPA in regard to the growth of human mammary carcinoma cells. PMID- 2974457 TI - Import pathways of precursor proteins into mitochondria: multiple receptor sites are followed by a common membrane insertion site. AB - The precursor of porin, a mitochondrial outer membrane protein, competes for the import of precursors destined for the three other mitochondrial compartments, including the Fe/S protein of the bc1-complex (intermembrane space), the ADP/ATP carrier (inner membrane), subunit 9 of the F0-ATPase (inner membrane), and subunit beta of the F1-ATPase (matrix). Competition occurs at the level of a common site at which precursors are inserted into the outer membrane. Protease sensitive binding sites, which act before the common insertion site, appear to be responsible for the specificity and selectivity of mitochondrial protein uptake. We suggest that distinct receptor proteins on the mitochondrial surface specifically recognize precursor proteins and transfer them to a general insertion protein component (GIP) in the outer membrane. Beyond GIP, the import pathways diverge, either to the outer membrane or to translocation contact-sites, and then subsequently to the other mitochondrial compartments. PMID- 2974458 TI - Immunocytochemistry of calciosomes in liver and pancreas. AB - Calciosomes are small cytoplasmic vacuoles identified in various nonmuscle cell types by their content of protein(s) similar to calsequestrin (CS), the Ca2+ storage protein of the muscle sarcoplasmic reticulum (SR). These entities have been interpreted as the "primitive" counterpart of the SR, and suggested to be the organelle target of inositol-1,4,5-triphosphate action (Volpe, P., K. H. Krause, S. Hashimoto, F. Zorzato, T. Pozzan, J. Meldolesi, and D. P. Lew. Proc. Natl. Acad. Sci. USA. 85:1091-1095). Immunoperoxidase and immunogold experiments carried out in both thick and ultrathin cryosections of rat hepatocytes and pancreatic acinar cells by using antimuscle CS antibodies revealed a specific labeling widely distributed in the entire cytoplasm, while nuclei were negative. Individual calciosomes appeared as small (105 nm) membrane-bound vacuoles intermingled with, and often apposed to ER cisternae and mitochondria. Other calciosomes were scattered in the Golgi area, in between zymogen granules and beneath the plasma membrane. The cumulative volume of the CS-positive organelles was measured to account for the 0.8 and 0.45% of the cytoplasm in liver and pancreas cells, respectively. The real total volume of the calciosome compartment is expected to be approximately twice as large. In hepatocytes, structures similar to CS-positive calciosomes were decorated by antibodies against the Ca2+ ATPase of muscle SR, while ER cisternae were not. By dual labeling, colocalization was revealed in 53.6% of the organelles, with 37.6% positive for the ATPase only. CS appeared preferentially confined to the content, and the Ca2+ ATPase to the contour of the organelle. The results suggested a partial segregation of the two antigens, reminiscent of their well-known segregation in muscle SR. Additional dual-label experiments demonstrated that hepatic calciosomes express neither two ER markers (cytochrome-P450 and NADH-cytochrome b5 reductase) nor the endolysosome marker, luminal acidity (revealed by 3-[2,4 dinitroanilino]-3'-amino-N-methyl dipropylamine). Calciosomes appear as unique cytological entities, ideally equipped to play a role in the rapid-scale control of the cytosolic-free Ca2+ in nonmuscle cells. PMID- 2974459 TI - Inhibition of kinesin-driven microtubule motility by monoclonal antibodies to kinesin heavy chains. AB - We have prepared and characterized seven mouse monoclonal antibodies (SUK 1-7) to the 130-kD heavy chain of sea urchin egg kinesin. On immunoblots, SUK 3 and SUK 4 cross-reacted with Drosophila embryo 116-kD heavy chains, and SUK 4, SUK 5, SUK 6, and SUK 7 bound to the 120-kD heavy chains of bovine brain kinesin. Three out of seven monoclonal antikinesins (SUK 4, SUK 6, and SUK 7) caused a dose dependent inhibition of sea urchin egg kinesin-induced microtubule translocation, whereas the other four monoclonal antibodies had no detectable effect on this motility. The inhibitory monoclonal antibodies (SUK 4, SUK 6, and SUK 7) appear to bind to spatially related sites on an ATP-sensitive microtubule binding 45-kD chymotryptic fragment of the 130-kD heavy chain, whereas SUK 2 binds to a spatially distinct site. None of the monoclonal antikinesins inhibited the microtubule activated MgATPase activity of kinesin, suggesting that SUK 4, SUK 6, and SUK 7 uncouple this MgATPase activity from motility. PMID- 2974460 TI - Fractionation of Tetrahymena ciliary membranes with triton X-114 and the identification of a ciliary membrane ATPase. AB - Cilia were isolated from Tetrahymena thermophila, extracted with Triton X-114, and the detergent-soluble membrane + matrix proteins separated into Triton X-114 aqueous and detergent phases. The aqueous phase polypeptides include a high molecular mass polypeptide previously identified as a membrane dynein, detergent soluble alpha and beta tubulins, and numerous polypeptides distinct from those found in axonemes. Integral membrane proteins partition into the detergent phase and include two major polypeptides of 58 and 50 kD, a 49-kD polypeptide, and 5 polypeptides in relatively minor amounts. The major detergent phase polypeptides are PAS-positive and are phosphorylated in vivo. A membrane-associated ATPase, distinct from the dynein-like protein, partitions into the Triton X-114 detergent phase and contains nearly 20% of the total ciliary ATPase activity. The ATPase requires Mg++ or Ca++ and is not inhibited by ouabain or vanadate. This procedure provides a gentle and rapid technique to separate integral membrane proteins from those that may be peripherally associated with the matrix or membrane. PMID- 2974461 TI - Benign and malignant forms of erythroderma: cutaneous immunophenotypic characteristics. AB - In order to determine if immunohistologic features are useful in distinguishing benign from malignant types of erythroderma, we studied the immunophenotype of lesional T cells in 20 patients (8 mycosis fungoides/Sezary syndrome, 12 benign) and found them to be generally similar. In all cases, the majority of T cells were Leu-1+, Leu-4+, and Leu-5+, as is typical of mature T cells. Although in most cases a majority of Leu-3+ (helper/phenotype) T cells were present, in 2 there was a majority of the Leu-2+ (cytotoxic/suppressor) subset and in 12 others, a significant minority (20%-40%) of these cells. Low percentages of Leu 2+ cells (less than or equal to 10%), resulting in high Leu-3+/Leu-2+ ratios, did not distinguish benign from malignant erythroderma. Leu-8 antigen deficiency was common in both mycosis fungoides/Sezary syndrome and benign cases (62% vs 75%, respectively). In contrast, Leu-9 antigen deficiency was present in only one patient in each group. The lack of combined Leu-8/9 antigen deficiency in our patients may be due to a heavy inflammatory T cell component, obscuring the antigen deficiencies seen in most nonerythrodermic mycosis fungoides cases. We conclude that immunophenotypic studies with the use of the current antibody panel show many similarities between benign and malignant forms of erythroderma, as well as some minor differences that may prove diagnostically useful if corroborated by future studies. PMID- 2974462 TI - Occupational dermatoses. PMID- 2974463 TI - Comparison of ipratropium solution, fenoterol solution, and their combination administered by nebulizer and face mask to children with acute asthma. AB - In a randomized, double-blind, parallel-group trial, 47 children with acute asthma received a combination of ipratropium bromide solution (250 micrograms) and fenoterol hydrobromide solution (625 micrograms), fenoterol solution (625 micrograms) alone, or ipratropium solution (250 micrograms) alone, administered by face mask and nebulizer, with the dose repeated 60 minutes later. The groups did not differ significantly with regard to age, pulmonary function at baseline, or any other variable. They were monitored at 30, 60, 90, and 120 minutes by use of a clinical score, oxygen saturation, and pulmonary function tests. At the end of the study, albuterol was administered to assess residual bronchoconstriction. Clinical scores improved significantly after treatment in all groups at all times compared with baseline. The greatest improvement in FEV1 was seen in the patients treated with ipratropium/fenoterol, whether considered as absolute change, change in percent predicted, or percent change from baseline. Ipratropium/fenoterol was significantly better than fenoterol alone only when considered as percent change from baseline. Improvement in flow at mid and low lung volumes was significantly greater for the ipratropium/fenoterol combination than for ipratropium alone; no significant differences were noted between ipratropium/fenoterol and fenoterol for flow at mid and low lung volumes. Treatment with albuterol did not significantly improve pulmonary function in the groups receiving ipratropium/fenoterol or fenoterol alone, but it did increase flow at all lung volumes in the group receiving ipratropium alone. No patient complained spontaneously of any adverse reactions, and no clinically significant changes in heart rate or systolic or diastolic blood pressures occurred.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974464 TI - Comparative study of collecting tubules and vasopressin binding capacity in the renal medulla of developing hypothyroid rat. AB - The effects of congenital hypothyroidism on both the structure and function of the renal medulla were studied by comparing, in 1-month old rats, the structural features of collecting tubules with the capacity of vasopressin to bind membrane preparations and the related adenylate cyclase activation. With the exception of a reduced caliber, hypothyroidism had no effect on the density, total number, distribution of tubules according to epithelial thickness, or on the number of epithelial cells, or their area. The binding capacity of vasopressin and the related adenylate cyclase activation were equally reduced by about 50%, without changes in (i) the basal or guanylyl-imidodiphosphate (Gpp(NH)p)-stimulated adenylate cyclase activities, (ii) the apparent dissociation constant (KD) of labelled vasopressin from its specific receptor or (iii) the apparent activation constant (Kact) of vasopressin for adenylate cyclase. Taken together, these results clearly demonstrate that congenital hypothyroidism exerts a direct influence on the developing responsiveness of the renal medulla, mainly by reducing the density of active hormone receptors per cell, instead of reducing cell number or cell membrane area. PMID- 2974465 TI - Plasma and urinary clearance rates of atrial natriuretic factor during ontogeny in sheep. AB - The present study was designed to determine the plasma clearance rate of atrial natriuretic factor (ANF) during development in chronically-instrumented fetal, newborn and adult non-pregnant sheep. To determine the contribution of the kidney in the metabolism of ANF, urinary clearance of ANF was also measured. Intravenous infusion of ANF (0.025 and 0.1 microgram.min-1.kg-1) produced a significant decrease in mean arterial blood pressure in newborn lambs and in adult non pregnant sheep. Estimated plasma ANF clearance rate for the 0.025 and 0.1 microgram.min-1.kg-1 ANF infusion rate were respectively 177 +/- 55 and 155 +/- 34 ml.min-1.kg-1 in fetuses, 138 +/- 26 and 97 +/- 13 ml.min-1.kg-1 in newborn lambs and, 148 +/- 33 and 103 +/- 25 ml.min-1.kg-1 in adult nonpregnant ewes. Fetal, newborn and adult ANF plasma clearance rates during high ANF infusion rate (0.1 microgram.min-1.kg-1) were not significantly different. Low or high ANF infusion rate was not associated with significant changes in urinary ANF concentration or urinary ANF excretion rate. Taken together, the present study demonstrates that ANF plasma clearance rate is similar in fetal, newborn and adult non-pregnant sheep and that the excretory function of the kidney contributes only minimally to ANF plasma clearance rate. PMID- 2974466 TI - The human immunodeficiency virus and its pathogenesis. AB - HIV is a lentivirus that infects many tissues of the body, including the cells of the brain, the hematopoietic system, and the bowel epithelium. Its spread is mediated by virus-infected cells. Control of HIV depends on a strong host immune response against virus-infected cells, either through suppression of virus release or through killing of the virus-infected cells. Present studies are directed at understanding mechanisms by which effective antiviral therapy and a vaccine can be developed. PMID- 2974467 TI - Comparison of results of recombinant and plasma-derived hepatitis B vaccines in Japanese nursery-school children. AB - Results obtained with a recombinant hepatitis B vaccine were compared with those obtained with a plasma-derived hepatitis B vaccine in separate studies conducted in nursery schools in which at least one child had hepatitis B e antigen associated with surface antigen. Recombinant vaccine (5 micrograms), made in Japan and prepared from antigen expressed in yeast, was given to 118 children (aged 0-5 years of age, mean age 2.9 years). Plasma-derived vaccine (10 micrograms) was given to 243 children. Side-reactions were not observed with either vaccine. Seroconversion rates for the recombinant vaccine group were 8.5% after 1 month, 98.3% after 9 months and 100% after 12 months. For the plasma derived vaccine group, the rate after 1 month was 26.3% after 9 months 82.3%, and after 12 months 77.9%. Although in the recombinant vaccine group the immune response developed more slowly during the early phase, seroconversion rates were significantly higher than in the plasma-derived vaccine group after 6 months. Titres of antibodies also were significantly higher in the recombinant vaccine group after the third injection. None of the children in either group became infected with hepatitis B virus. These results confirm the high immunogenicity, safety and efficacy of the recombinant vaccine given to these nursery school children. PMID- 2974468 TI - Protection against acute lethal viral infections with the native steroid dehydroepiandrosterone (DHEA). AB - A significant protective effect of a native adrenal steroid, dehydroepiandrosterone (DHEA), was demonstrated in studies of two lethal viral infection models in mice: systemic coxsackievirus B4 and herpes simplex type 2 encephalitis. The steroid was active either by long-term feeding or by a single subcutaneous injection. A closely related steroid, etiocholanolone, was not protective in these models. Histopathological analysis, leukocyte counts, and numbers of spleen antibody forming cells in the coxsackievirus B4 model suggests that DHEA functions by maintaining or potentiating the immune competence of mice otherwise depressed by viral infection. DHEA was not effective in genetically immunodeficient HRS/J hr/hr mice and did not demonstrate antiviral activity in vitro. While the molecular basis for DHEA's effect on the immune system is not known, studies by others suggest that it may counteract the stress related immunosuppressive effects of glucocorticoids stimulated by viral infection. Because DHEA is a native steroid that has been used clinically with minimal side effects, the utility of DHEA in the therapeutic modulation of acute and chronic viral infections including the acquired immune deficiency syndrome deserves intensive study. PMID- 2974469 TI - Isozyme variation in Simulium (Edwardsellum) damnosum s.l. (Diptera: Simuliidae) from Kenya. AB - Isozyme variation of the Simulium damnosum sibling species complex was studied by cellulose acetate electrophoresis (CAE) from four Kenyan river systems. Two enzymes, PGM and HK, were diagnostic and differentiated the larvae collected in Western and Nyanza provinces from the larvae collected at Mt. Kenya. Allele frequency differences of the enzyme PGI allowed about 75% separation of the geographically distinct populations. PMID- 2974470 TI - First results of the application of isozyme analysis to study of Culicoides (Diptera: Ceratopogonidae). AB - Results of a biochemical study on the genus Culicoides are reported. A method of analysis using six discriminant enzymatic systems was used and the results obtained on zymograms are detailed. A genetic study of C. nubeculosus, based on phosphoglucomutase was carried out. The processing of the data obtained, by means of correspondence analysis, provides an interesting comparative study between morphologically related species. PMID- 2974471 TI - T cell subsets and disease progression after total lymphoid irradiation in chronic progressive multiple sclerosis. AB - T lymphocyte subset percentages were determined in 16 total lymphoid irradiation (TLI) treated and 18 sham treated control patients with chronic progressive multiple sclerosis. During the first year after treatment, the ratio of T helper/inducer to T suppressor/cytotoxic cells (Th/Ts ratio) was significantly higher in sham treated multiple sclerosis patients who worsened clinically compared with TLI treated and sham treated multiple sclerosis patients who remained clinically stable. TLI caused a fall in the percentage of T helper cells in treated patients, while the percentage of T suppressor cells remained stable during the first year after treatment. In contrast, the percentage of T suppressor cells fell in sham treated multiple sclerosis patients who worsened clinically. PMID- 2974472 TI - The proportion of suppressor-inducer T-lymphocytes is reduced in recurrent aphthous stomatitis. AB - A flow cytometric analysis of peripheral blood lymphocytes was undertaken in recurrent aphthous stomatitis patients. The project aimed at detecting differences within lymphocyte subsets using type-specific monoclonal antibodies. Peripheral blood samples were taken from RAS patients in both active and remission phases of the disease and from a group of healthy control subjects. There were no statistical differences between the active and remission phases within any of the lymphocyte subsets examined. There was, however, a significant difference between the RAS group and the control group. RAS patients have depressed CD4+ cell numbers and elevated CD8+ cell numbers. The CD4:CD8 ratio is also depressed. A dissection of the CD4+ subset shows raised numbers of CD4+, 4B4+ lymphocytes and depressed numbers of CD4+, 2H4+ lymphocytes. Previous studies have shown disruption of peripheral blood lymphocyte numbers in Behcet's syndrome. A similar pattern has now been shown in uncomplicated cases of minor RAS. PMID- 2974473 TI - Distribution of mucopolysaccharide in site of initial intramembranous osteogenesis in fetal mouse mandible. PMID- 2974474 TI - Effect of minocycline on prostaglandin formation in gingival fibroblasts. PMID- 2974475 TI - Periodontal ligament fibroblasts, preosteoblasts, and prechondrocytes express receptors for epidermal growth factor in vivo: a comparative radioautographic study. PMID- 2974476 TI - In situ activated T lymphocytes in active versus stable periodontal lesions. PMID- 2974477 TI - Effects of race and periodontal status on antibody reactive with Actinobacillus actinomycetemcomitans strain Y4. PMID- 2974478 TI - Production of chemotactic factors for neutrophils following the interaction of Bacteroides gingivalis with purified C5. PMID- 2974479 TI - Effect of collagen solution application on healing following surgical treatment in colony-bred monkeys. PMID- 2974480 TI - The effect of patient-associated factors on the reproducibility of the mouth bleeding score. PMID- 2974481 TI - Blastogenic response and immunoglobulin production by inflamed gingival lymphocytes from dogs. PMID- 2974482 TI - Purification and characterization of Eikenella corrodens aggregating factor from submandibular-sublingual saliva. PMID- 2974483 TI - The action of sodium fluoride on suspected periodontopathogens. PMID- 2974484 TI - On the use of Good's L-statistic in the analysis of bacteriologic samples. PMID- 2974485 TI - Comparisons of descending pain inhibitory pathways activated by beta-endorphin and morphine as characterized by supraspinal and spinal antinociceptive interactions in mice. AB - Morphine administered concurrently by i.c.v. plus intrathecal (i.t.) injection produces a multiplicative (synergistic) interaction for antinociception in the tail-flick test. Inasmuch as i.c.v. administered beta-endorphin has been proposed to produce antinociception by activating a descending pain inhibitory system different from that activated by morphine, the present experiments compared the two systems in mice. The responses to i.c.v., i.t. and combinations of i.c.v. plus i.t. administration of morphine and beta-endorphin were evaluated by determination of ED50 values which were plotted as isobolograms and compared to calculated theoretical additive ED50 values. The following combinations gave additive interactions: i.c.v. plus i.t. beta-endorphin, i.c.v. beta-endorphin plus i.t. morphine and i.t. morphine plus i.t. beta-endorphin. These results were consistent with the hypothesis that i.c.v. beta-endorphin stimulates supraspinal epsilon receptors which activate a descending pathway involving enkephalinergic neuronal mediation and spinal postsynaptic mu receptors. Stimulation of these mu receptors by i.t. morphine or i.t. beta-endorphin together with the supraspinal effect of beta-endorphin resulted in an additive interaction. Multiplicative interactions were obtained for the following combinations: i.c.v. morphine plus i.t. morphine, i.c.v. morphine plus i.t. beta-endorphin and i.c.v. morphine plus i.c.v. beta-endorphin. Morphine administered i.c.v. stimulated supraspinal mu receptors to activate a descending pain inhibitory pathway which is mediated spinally by monoamines. The i.t. agonists in this case activated the spinal mu receptor which is presumed to be part of the beta-endorphin descending pathway described above. Thus, when both pathways were activated simultaneously the interaction was multiplicative.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974486 TI - Rapid discrimination of the stimulus properties of 5-hydroxytryptamine agonists using conditioned taste aversion. AB - Separate groups of rats were trained to discriminate the stimulus properties of selective agonists at 5-HT receptors using a conditioned taste aversion procedure. Fluid-restricted rats were injected with drug or saline and then given access to a 0.25% saccharin solution for 30 min. When rats received a drug trial, saccharin consumption was followed by an injection of LiCl (1.8 mEq/kg i.p.), whereas on saline trials saccharin consumption was followed by a second injection of saline instead of LiCl. Rats were trained using injections of either 8-hydroxy 2-(di-n-propylamino)tetralin (0.4 mg/kg i.p.), an agonist selective for the 5 HT1A receptor, or 1-(m-trifluoromethylphenyl)piperazine (0.8 mg/kg i.p.), an agonist selective for 5-HT1B and 5-HT1C receptors, as the drug stimuli. Acquisition of the discriminated taste aversion, as measured by the differential effects on saccharin drinking between drug and saline trials, required only two to three pairings of either drug stimulus with LiCl injections. The 8-hydroxy-2 (di-n-propylamino)tetralin discriminative stimulus cue generalized to other drugs that are selective for the 5-HT1A receptor, such as ipsapirone (8-16 mg/kg i.p.) or buspirone (4 mg/kg i.p.), but not to agonists that are selective for the 5 HT1B/1C receptor, such as 1-(m-trifluoromethylphenyl)piperazine or 1-(m chlorophenyl)piperazine. The discriminative stimulus properties of 1-(m trifluoromethylphenyl)piperazine generalized to 1-(m-chlorophenyl)piperazine (0.2 0.8 mg/kg i.p.) but not to the 5-HT1A agonist 8-hydroxy-2-(di-n propylamino)tetralin (0.4 mg/kg i.p.).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974487 TI - Desensitization of platelet thromboxane A2/prostaglandin H2 receptors by the mimetic U46619. AB - Thromboxane A2 (TXA2) and prostaglandin H2 (PGH2) activate platelets through membrane receptors. This study sought to determine if changes occur in the TXA2/PGH2 receptor during its desensitization induced by exposure to its agonist 11 alpha,9 alpha-(epoxymethano)prosta-5Z,13E-dienoic acid (U46619). Washed human platelets were incubated for 30 min with U46619 in the presence of an antiaggregatory agent, Iloprost. The platelets were washed and the aggregation response to U46619 was determined. The EC50 increased from 372 +/- 94 nM in the control group to 826 +/- 143 nM for the U46619-pretreated group (n = 7). This desensitization was specific inasmuch as the aggregation responses to thrombin and the calcium ionophore A23187 were not affected by U46619 pretreatment. Desensitization was accompanied by a decrease in the number of binding sites for [3H]U46619 from 789 +/- 189 in the control to 386 +/- 120 sites per platelet in the U46619-treated group (n = 5) and a decrease in the number of binding sites for the TXA2/PGH2 receptor antagonist, [125I]9,11-dimethylmethano-11,12-methano 16(3-iodo-4-hydroxyphenyl )-13,14- dihydro-13-aza-15-alpha beta-omega-tetranor TXA2 from 3988 +/- 957 to 2443 +/- 553 (n = 8). The Kd for U46619 was 37 +/- 10 nM in the control group and 23 +/- 11 nM for the U46619-treated group (n = 5). The Kd for I-9,11-dimethylmethano-11,12-methano-16(3-iodo-4-hydroxyphenyl)-13 ,14 dihydro- 13-aza-15-alpha beta-omega-tetranor-TXA2 changed from 58 +/- 12 nM in the control to 44 +/- 9 nM in the U46619-treated group (n = 8).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974489 TI - Psychosocial predictors of disability in patients with low back pain. AB - We prospectively studied outcomes for 179 patients with low back pain. Predictors of future function, employment, and medical utilization were drawn from 21 clinical, demographic, and psychosocial variables using multivariate techniques. Education, previous episodes, and whether the patient "always feels sick" were independently associated with most outcome measures, but prescribed therapy and physical findings were not. These 3 items created a scale defining subgroups with 3-fold differences in outcomes (e.g., 35% functionally improved in the worst group vs 93% in the best, p less than 0.001). Data from a national survey supported the importance of education and self-rated health as correlates of back related disability. PMID- 2974488 TI - Immunoregulatory defects in juvenile rheumatoid arthritis. Comparison between patients with the systemic or polyarticular forms. AB - Studies of cellular immunity in juvenile chronic arthritis (juvenile rheumatoid arthritis, JRA) have been scant, controversial, or have not addressed the issue of the different forms of the disease. We studied 23 patients with JRA of either systemic (n = 8) or polyarticular (n = 15) type of onset and compared the findings to those made in 10 healthy children of similar age. Both groups of patients with JRA were found to have increased CD8 T cells, normal production of interleukin-1 and 2 and decreased production of B cell stimulatory factor in their peripheral blood. In addition, patients with systemic JRA were found to have decreased spontaneously expanded and concanavalin-A induced suppressor functions. These findings in both forms of JRA are distinguishable from those that have been made in other connective tissue diseases including the adult form of rheumatoid arthritis. PMID- 2974490 TI - Idiopathic retroperitoneal fibrosis presenting with back pain. AB - We describe 2 cases of idiopathic retroperitoneal fibrosis presenting with back pain. Additional features included weight loss, constitutional symptoms and elevated erythrocyte sedimentation rate (ESR). One patient had sacroiliitis and was HLA-B27 positive. A further finding was obstruction of the superior mesenteric artery which has not been previously described in retroperitoneal fibrosis. The importance of considering idiopathic retroperitoneal fibrosis in the differential diagnosis of back pain has not been emphasized in the rheumatological literature. PMID- 2974491 TI - Rheumatoid arthritis with T cell subsets suggesting chronic viral infection. PMID- 2974492 TI - [Experimental study of the treatment of uterine leiomyosarcoma in the mouse with progestogen]. AB - Because of its rare occurrence in the human, the endocrinologic and receptor related aspects of an uterine leiomyosarcoma (LMS) are poorly understood when compared to what is known of, say, human endometrial cancer. Thus, to increase our understanding, we have succeeded, by the string method, in inducing an uterine LMS in the mouse and have studied the possibility of hormonal therapy as a method of treatment. The findings of our study are enumerated as follows: 1. The induced uterine LMS had an estrogen receptor, which was confirmed by a biochemical assay and, morphologically, by a PAP (the peroxidase anti-peroxidase technique); 2. The growth of this tumor was significantly inhibited by MPA (medroxyprogesterone acetate) therapy (100 mg/kg); 3. After MPA therapy, the estrogen receptor levels were increased, especially in the nucleus; and, 4. The growth of a secondary tumor, transplanted after the initial hormone therapy, was not inhibited by the readministration of MPA. Our results suggest that this experimentally-induced uterine LMS in the mouse provides a useful means to study therapeutic treatment, and may assist in furthering our understanding of human uterine LMS and lead to finding an effective therapy. PMID- 2974493 TI - [Megakaryoblastic leukemia and transient abnormal myelopoiesis with Down's syndrome]. PMID- 2974494 TI - Age-associated alteration in actomyosin ATPase activity of human myocardium--I. Calcium sensitivity of Mg2+-ATPase activity. PMID- 2974495 TI - [Ainu midwifery technics practiced by Ms Aiko. 19. Backache during and after delivery]. PMID- 2974496 TI - [Ainu midwifery technics practiced by Ms. Aiko. 18. Backache during and after delivery. II. Diagnosis and treatment]. PMID- 2974497 TI - [Oral and lip training of severely handicapped children using hard candies]. PMID- 2974498 TI - [Regional health care activities and expectations on public health nursing: primary care and normalization (a state of having a normal percentage of the disabled within the local population)]. PMID- 2974499 TI - Angioscopy as an adjunct to laser-assisted angioplasty in seven peripheral vascular cases. PMID- 2974500 TI - [Physical and social disability among the aging]. PMID- 2974501 TI - [Early diagnosis of cardiovascular diseases using expert personal computer systems]. PMID- 2974502 TI - [Arterial pressure reaction to physical load in patients with Leriche's syndrome and right-ventricular myocardial hypertrophy]. PMID- 2974503 TI - [A successful treatment of a penetrating wound of the heart in a Central Regional Hospital]. PMID- 2974504 TI - Surrogate motherhood and the best interests of children. PMID- 2974506 TI - [The radiologic service of the Cheliabinsk Oblast (history, present status, future development)]. AB - Since 1939 the radiological service in the Chelyabinsk Region has developed into a well-organized system, fitted out with up-to-date equipment capable of providing all kinds of radiological care to cancer patients using modern therapeutic methods. PMID- 2974505 TI - Low-back pain--diagnosis, treatment, and medicolegal considerations. PMID- 2974507 TI - [Pulmonary hypertension and Sjogren's syndrome]. PMID- 2974508 TI - Blood transfusion in renal transplantation--the induction of tolerance by incompatibility for class I antigen. AB - Blood transfusion given before renal transplantation has been shown to have a powerful immune modulating effect on recipient response to kidney allograft. The mechanism responsible for this effect is still unknown. Here it is assumed that, due to incompatibility for HLA-A related class I antigens between blood donor and recipient, pretransplant blood transfusion may allospecifically induce in the recipient the generation of T cells that have a suppressive effect on T lymphocyte response to incompatible HLA class II antigens present on kidney allograft. Although in many respects this interpretation is still speculative, it is in accord with findings reported from clinical and experimental studies of the effect on graft survival of blood transfusion, both random and donor specific. PMID- 2974509 TI - [Work posture and spinal changes in porphyry miners]. PMID- 2974510 TI - Is alcohol to blame? The story of Hluphekile. PMID- 2974513 TI - [Spontaneous hematoma of the rectus abdominis muscle]. PMID- 2974511 TI - Cloning and expression of a rat D2 dopamine receptor cDNA. AB - Dopamine receptors are classified into D1 and D2 subtypes on the basis of their pharmacological and biochemical characteristics. The D2 dopamine receptor has been implicated in the pathophysiology and treatment of movement disorders, schizophrenia and drug addiction. The D2 dopamine receptor interacts with guanine nucleotide-binding proteins to induce second messenger systems. Other members of the family of receptors that are coupled to G proteins share a significant similarity in primary amino-acid sequence and exhibit an archetypical topology predicted to consist of seven putative transmembrane domains. We have taken advantage of the expected nucleotide sequence similarities among members of this gene family to isolate genes coding for new receptors. Using the hamster beta 2 adrenergic receptor gene as a hybridization probe we have isolated related genes including a cDNA encoding the rat D2 dopamine receptor. This receptor has been characterized on the basis of three criteria: the deduced amino-acid sequence which reveals that it is a member of the family of G-protein-coupled receptors; the tissue distribution of the mRNA which parallels that of the D2 dopamine receptor; and the pharmacological profile of mouse fibroblast cells transfected with the cDNA. PMID- 2974512 TI - Transformation of the Fusarium mycotoxin zearalenone in maize cell suspension cultures. PMID- 2974514 TI - [Laparoscopy and ovarian cancer]. PMID- 2974515 TI - [Blocking factors in pregnancy. Review of the literature and a case series]. PMID- 2974516 TI - Effects of chloralose-urethane anesthesia on single-axon reciprocal Ia IPSPs in the cat. AB - Reciprocal Ia inhibitory postsynaptic potentials (IPSPs) generated by single afferents have been recorded with signal averaging in unanesthetized ischemic decapitate cats for comparison with measurements previously obtained from preparations anesthetized with a mixture of chloralose and urethane. The results are similar to those which we obtained recently for single-axon recurrent IPSPs. Together, the studies show that chloralose-urethane anesthesia has a depressant effect on two widely studied circuits in the mammalian spinal cord. PMID- 2974517 TI - Hepatitis B vaccines. PMID- 2974518 TI - The treatment of the homebound patient. PMID- 2974519 TI - Increase in reimbursement should improve nursing home care. PMID- 2974520 TI - Physiotherapy and back pain. PMID- 2974521 TI - [The visual functions of workers at the converter shop of the Karaganda Metallurgical Combine from an ophthalmic ergonomic viewpoint]. PMID- 2974522 TI - [Pathomorphology of the eyes in Down's disease]. PMID- 2974523 TI - [Dispensary care of children with ophthalmologic pathology in Turkmenistan]. PMID- 2974524 TI - [The potentials for optimizing the treatment process at a small eye department of a medical unit]. PMID- 2974525 TI - Hematologic and surgical management of the dental patient with plasminogen activator deficiency. AB - Anticoagulation therapy is used to treat patients with a variety of hemostatic disorders in an attempt to prevent thrombus formation. A thorough understanding of the patient's medical history is essential before dental treatment that may require alteration of this anticoagulation therapy. Alteration of anticoagulation therapy should be undertaken only after consultation with the patient's physician because some patients are at greater risk than others for thrombus formation or hemorrhage. This case of a 29-year-old man with plasminogen activator deficiency illustrates how consultation can result in a coordinated treatment plan for medical and dental management formulated to help ensure safe surgical treatment for these medically compromised patients. PMID- 2974526 TI - [Nuclear cardiologic examinations in patients on disability compensation after myocardial infarct]. PMID- 2974527 TI - A threshold effect in the induction of tumorigenicity of an established human cell line by v-mos. AB - The nontumorigenic immortal human cell line, SV80, was transfected with the v-mos gene to assess the gene's effect on tumorigenicity of cultured human cells. Two classes of cells, each containing functional v-mos, were obtained. The first class contained low levels of v-mos RNA, was morphologically transformed, but was nontumorigenic in nude mice. The second was also morphologically transformed, but contained high levels of v-mos RNA and was tumorigenic. The results indicate that SV80 cells behave similarly to murine fibroblasts in their response to v-mos in that they can be rendered tumorigenic by the viral oncogene. However, tumorigenicity was effected through a mechanism which involves different threshold doses for morphologic and tumorigenic transformation. PMID- 2974528 TI - E1A products of adenoviruses reduce the expression of cellular proliferation associated genes. AB - Products of the adenovirus (Ad) E1A region are capable of modulating the expression of a variety of genes, both of viral and cellular origin. We have investigated the expression of two proliferation-associated genes, c-myc and JE, in various adenovirus-transformed cell lines. Rodent and human cell lines, transformed as primary cultures by the E1 region of Ad5 or of Ad12, exhibit a markedly reduced expression of both genes. Ad5 E1-transformed cells show a more pronounced reduction of c-myc RNA than Ad12 E1-transformed cells. Even the very low levels of E1A products in Ad5 E1A-immortalized cells are sufficient for this reduction. Analysis of c-myc and JE expression in cell lines, transformed by other viral or cellular oncogenes, indicated that this reduction is specific for adenoviruses. These observations show that E1A does not contribute to cellular transformation by activating the proliferation-associated genes c-myc and JE, but instead suggest an alternative mechanism of E1A-mediated transformation. PMID- 2974529 TI - [A new species of black flies Psilocnetha almae sp. n. (Simuliidae) from southern Kazakhstan]. PMID- 2974530 TI - Multiple sulfatase deficiency: bridge between neuronal storage diseases and leukodystrophies. AB - A fatal case of multiple sulfatase deficiency in a 10-year-old girl is reported. In this rare disease, which is inherited as an autosomal recessive, features of metachromatic leukodystrophy and of mucopolysaccharidoses occur together. The white matter suffers progressive destruction with sulfatides accumulating in macrophages; these stain metachromatically as golden brown granules with acetic acid thionin stain. Cortical and subcortical neurons are distended by lipids and mucopolysaccharides, and fibrous thickening of the leptomeninges leads to severe obstructive hydrocephalus. This entity can be regarded as a bridge between leukodystrophies and neuronal storage diseases both conceptually and in its morphological manifestations. PMID- 2974531 TI - Oligoclonal T cell proliferative disorder in combined immunodeficiency. AB - Oligoclonal lymphoid proliferations may occur in immunocompromised patients and in the elderly. So far these proliferations have been shown to be of B cell origin. We describe a patient with a combined immunodeficiency, characterized by profound hypogammaglobulinemia and the initial absence of T lymphocytes in the peripheral blood (PB). From the age of 4 yr CD3+ T cells appeared in PB in rising numbers. These cells mainly expressed the CD4-/CD8+ phenotype (CD4/CD8 ratio: 0.1). Despite the emergence of T lymphocytes no proliferation of PB mononuclear cells could be induced with phytohemagglutinin, concanavalin A, or pokeweed mitogen. Between the ages of 4 and 6 yr the patient gradually developed hepatosplenomegaly and an interstitial pulmonary infiltrate of unknown origin, necessitating biopsies of both liver and lung. Infiltrates consisting of CD8+ T lymphocytes were found in the liver as well as the lung. CD8+ T cells were also abundant in the bronchoalveolar lavage fluid. Southern blot analysis of mononuclear cells from PB and of a lung biopsy specimen was performed to investigate if a clonal T cell population was involved. Analysis of the T cell receptor beta genes revealed that at least three expanded T cell clones were present in PB, one of which had invaded the lung. Thus far, i.e. 2 yr after the initial detection of clonal T cell receptor beta gene rearrangements, there have been no clinical or histologic signs of malignant behavior. We conclude that this combined immunodeficiency patient has a benign oligoclonal T cell lymphoproliferative disorder. Similar proliferations might well occur in other immunodeficiency states, whether primary or acquired. PMID- 2974532 TI - Fluorescent cytometric analysis of polymorphonuclear leukocytes in Chediak Higashi syndrome: diminished C3bi receptor expression (OKM1) with normal granular cell density. AB - Chediak-Higashi Syndrome (CHS) has been associated with recurrent bacterial infections and defective polymorphonuclear (PMN) leukocyte function. Confirmation of the diagnosis of CHS and defective PMN function was established in a 2-month old with accelerated phase CHS. The diagnosis was confirmed by demonstrating reduced PMN degranulation (beta-glucuronidase release 34.1 +/- 0.9% versus 5.1 +/ 4% and lysozyme release 17.6 +/- 1.2% versus 11.1 +/- 7% (control versus CHS) and staphylococcal bacterial killing at 15' 51.4 +/- 3.6% versus 24.9 +/- .4% (control versus CHS). Additional studies using fluorescent cytometric analysis were made to investigate other etiologies of PMN dysfunction in CHS. Total cell density and PMN granularity, as measured by fluorescent-activated cell sorter side scatter analysis, was no different from CHS and age-matched controls. Although CHS is characterized by large PMN granular inclusions, right angle light scatter analysis in this study suggests that the total cell density within the PMN of patients with CHS is normal (D less than .01). PMN granular release of surface receptors was also studied using antibody binding and fluorescent analysis. OKM1 antibody-binding demonstrated significantly reduced C3bi (MO-1) receptor expression (13% of control) p less than 0.001. Decreased surface reception expression of C3bi receptors may play an additional role in defective PMN mobility, chemotaxis, and bactericidal activity in patients with CHS. PMID- 2974533 TI - [Distribution of antigens of the HLA system in children with primary arterial hypertension living in the Kuznetsk region]. PMID- 2974534 TI - [Organization of preventive services in a pediatric polyclinic and a consultation center for women in obstetric-therapeutic-pediatric units]. PMID- 2974536 TI - The complete primary structure of the human snRNP E protein. AB - The snRNP E protein is one of four "core" proteins associated with the snRNAs of the U family (U1,U2,U4,U5, and U6). Screening of a human teratoma cDNA library with a partial cDNA for a human autoimmune antigen resulted in the isolation of a cDNA clone containing the entire coding region of this snRNP core protein. Comparison of the 5' end of this cDNA with the sequences of two processed pseudogenes and primer extension data suggest that the cDNA is nearly full length. The longest open reading frame in this clone codes for a basic 92 amino acid protein which is in perfect agreement with amino acid sequence data obtained from purified E protein. The predicted sequence of this protein reveals no extensive similarity to other snRNP proteins, but contains regions of similarity to a eukaryotic ribosomal protein. PMID- 2974535 TI - An in vitro interaction between the human U3 snRNP and 28S rRNA sequences near the alpha-sarcin site. AB - Model transcripts containing mammalian pre-rRNA sequences were incubated with a HeLa cell extract, digested with T1 RNase, and immunoprecipitated with anti (U3)RNP or control antibodies. Two overlapping fragments derived from the 3' domain of human 28S rRNA were specifically immunoprecipitated although transcripts which spanned the transcription initiation site, the ETS processing site, the 5' end of 18S, and both termini of 5.8S yielded no protected fragments. The sequence of these fragments was determined using a novel technique in which the [32P]-labeled fragment was co-finger-printed with [3H]-labeled total transcript serving as an internal marker. The fragments immunoprecipitated derive from nucleotides 4570-4590 and 4575-4590 of human 28S and are adjacent to the alpha-sarcin site. Protection most likely involves the U3 RNA since it is sensitive to pretreatment of the extract with micrococcal nuclease. Complementarity between U3 and this rRNA region is phylogenetically conserved in species ranging from human to S. cerevisiae. The possible significance of this finding is discussed. PMID- 2974537 TI - Effects of palindromes on in vivo DNA replication and mutagenesis in bacteriophage phi X174 RF DNA. AB - Bacteriophage phi X174 mutants with insertions of palindromic DNA sequences are rapidly outgrown by competing wild-type phage (Muller & Turnage, J. Mol. Biol. 189: 285). The basis for this defect was investigated and found to be due to an exclusion event early in the infectious cycle, in which phage genomes with palindromic inserts were preferentially excluded by wt phage. In addition, we have obtained further evidence for a palindrome induced genetic instability. Both defects are dependent on palindrome size and sequence, consistent with a model which involves formation of cruciforms, or cruciform-like structures. We propose that formation of unusual DNA secondary structures reduces the effectiveness of replicative form (RF) DNA to interact with limiting replication factors or membrane binding sites, possibly because of interaction with the host recombination system. PMID- 2974538 TI - Involvement of a cryptic ATPase activity of UvrB and its proteolysis product, UvrB* in DNA repair. AB - The incision of damaged DNA by the Escherichia coli UvrABC endonuclease requires ATP hydrolysis. Although the deduced sequence of the UvrB protein suggests a putative ATP binding site, no nucleoside triphosphatase activity is demonstrable with the purified UvrB protein. The UvrB protein is specifically proteolyzed in E. coli cell extracts to yield a 70 kD fragment, referred to as UvrB*, which has been purified and is shown to possess a single-strand DNA dependent ATPase activity. Substrate specificity and kinetic analyses of UvrB* catalyzed nucleotide hydrolysis indicate that the stimulation in DNA dependent ATPase activity following formation of the UvrAB complex results from the activation of the normally sequestered UvrB associated ATPase. Using nucleotide analogues, it can be shown that this activity is essential to the DNA incision reaction carried out by the UvrABC complex. PMID- 2974539 TI - Rapid isolation of lambda phage DNA in micro- and macro-variants. PMID- 2974541 TI - A fair deal for the disabled. PMID- 2974540 TI - Direct cross-linking of snRNP proteins F and 70K to snRNAs by ultra-violet radiation in situ. AB - Protein-RNA interactions in small nuclear ribonucleoproteins (UsnRNPs) from HeLa cells were investigated by irradiation of purified nucleoplasmic snRNPs U1 to U6 with UV light at 254 nm. The cross-linked proteins were analyzed on one- and two dimensional gel electrophoresis systems, and the existence of a stable cross linkage was demonstrated by isolating protein-oligonucleotide complexes from snRNPs containing 32P-labelled snRNAs after exhaustive digestion with a mixture of RNases of different specificities. The primary target of the UV-light induced cross-linking reaction between protein and RNA was protein F. It was also found to be cross-linked to U1 snRNA in purified U1 snRNPs. Protein F is known to be one of the common snRNP proteins, which together with D, E and G protect a 15-25 nucleotide long stretch of snRNAs U1, U2, U4 and U5, the so-called domain A or Sm binding site against nuclease digestion (Liautard et al., 1982). It is therefore likely that the core-protein may bind directly and specifically to the common snRNA domain A, or else to a sub-region of this. The second protein which was demonstrated to be cross-linked to snRNA was the U1 specific protein 70K. Since it has been shown that binding of protein 70K to U1 RNP requires the presence of the 5' stem and loop of U1 RNA (Hamm et al., 1987) it is likely that the 70K protein directly interacts with a sub-region of the first stem loop structure. PMID- 2974542 TI - [Risk factors for female osteoporosis]. PMID- 2974543 TI - [Detection of female osteoporosis]. PMID- 2974544 TI - [Evaluation using DSM-III of the clinical activity of a psychiatry department at a general hospital]. AB - The DSM-III multiaxial classification was used to evaluate the one-year clinical activity of a psychiatry and medical psychology department in a general hospital. Compared to the general population, the patients (n = 1,071) corresponded to a sample of professionally active subjects in which top and middle executives as well as students were over=represented. Fifty-one per cent of the patients suffered from anxiety or affective disorders, while 9 per cent presented somatoform disorders. Histrionic and dependent personalities were more frequent in women. Thirty six per cent of the subjects had a concurrent physical illness. Psychosocial stress and altered adaptation were higher in unemployed people and in patients with physical illness. Specific diagnostic and socioprofessional patterns corresponded to the various types of mental care: psychoanalytically oriented psychotherapy, behaviour therapy, psychopharmacological treatments, consultation-liaison, pain consultation. PMID- 2974545 TI - [Abdominal and retroperitoneal masses. Diagnostic value of fine needle cytopunctures. 121 consecutive controlled cases]. AB - A series of 121 consecutive patients with abdominal (96 cases) and retroperitoneal (25 cases) masses was studied in order to evaluate the contribution of cytology in establishing a definite diagnosis of the tumors. The cytologic results on aspirates were obtained with 22 or 23 gauge Chiba needles. At the same time, percutaneous fine needle aspiration biopsy was performed with 19 or 21 gauge fine needles having circumferentially bevelled tips that produced tiny tissue cores suitable for histologic study in 88 cases. These diagnoses were controlled with larger pathological specimens in 78 cases and appeared to be consistent with the clinical and biological course in 43 cases. The sensitivity of the cytologic diagnosis was 83 per cent, and its overall accuracy 87.6 per cent. There were no false-positive results. Furthermore, the method proved to be safe and well-tolerated, with low morbidity (0.8 per cent) and no mortality. PMID- 2974546 TI - [Infectious complications, delayed wound healing and inflammatory syndromes as late complications of cardiovascular surgery]. AB - Four-hundred and three patients were admitted to an internal medicine unit for late complications of cardiac or vascular surgery; 321 of these patients (80 per cent) had a total of 379 complications or intercurrent disease, 147 (40 per cent) of which were infections (71 cases), delayed healing of the operative wound (39 cases) or inflammatory syndromes (37 cases). Such complications are notable for their immediate or secondary severity, the diagnostic problems sometimes raised by their latency, their repercussions on the length of hospitalization and cost of surgery, and the thoughts they inspire concerning the minimal stay in hospital required after this type of surgery. PMID- 2974547 TI - [Hydatid cyst of the pancreas responsible for chronic recurrent pancreatitis]. AB - Hydatid cyst of the pancreas is a rare localization of hydatidosis. It raises diagnostic problems which can only be solved by section of the surgical specimen. An exceptional case of hydatid cyst of the pancreatic tail which resulted in chronic recurrent pancreatitis, and a review of the literature, enable to put hydatidosis on the list of causes of pancreatitis. PMID- 2974548 TI - [Doppler effect in cardiology. Continuous Doppler, pulsed Doppler, Doppler color]. AB - Echocardiography has contributed to the exploration of the heart by providing much information, and it has even given rise to new semiological concepts. However, abnormalities of intracardial blood flow, notably shunts and regurgitations, could only be diagnosed indirectly from their effects on cardiac cavities. A new step forward was the advent of pulsed and continuous doppler ultrasound, since from that moment it has been possible to demonstrate abnormal blood flows, to measure their velocity and to determine, albeit with some reservation, such crucial values as pressure gradients. Yet even when these two techniques were combined in the doppler-echotomography systems blind areas persisted, and a blood flow of strongly abnormal direction could in fact escape doppler velocimetry. This is where another development came to the rescue, for it enabled both normal and abnormal flows to be visualized in colours. Owing to this visualization, and provided all possible projections are used, it has become exceptional to "miss" an abnormal blood flow. The diagnosis is now easier and more accurate, and in a second stage the flow can be quantified by pulsed or continuous doppler ultrasound. The colour-coded doppler technique therefore has not superseded the previous one: one may say that it has merely increased diagnostic reliability, but is this not a decisive improvement? PMID- 2974549 TI - [Surgical treatment of female stress urinary incontinence. Determination of the location of the vesico-urethral junction]. AB - The accuracy of the various methods used to locate the vesico-urethral junction was evaluated in 20 female patients. In each patient the following parameters were successively measured: functional length of the urethra (urethral profile); distance from the meatus to the posterior lip of the vesical cervix (endoscopy), length of the catheter segment between the balloon and the meatus. Finally, the location of the balloon was determined intra-operatively and compared with the "urethroscopic" findings. The results obtained showed that locating the vesico urethral junction solely from the location of the balloon could be misleading and that there was a significant difference (nearly 8 mm) between functional and urethroscopic measurements. These results led to a mathematical formula which enables the anatomical length of the urethra to be measured from its functional length. The authors suggest that this length, measured with an urethrometer, should be used for precise location of the junction during surgery for female stress incontinence. PMID- 2974550 TI - [Seropositivity for the human immunodeficiency virus in systemic lupus erythematosus]. PMID- 2974551 TI - [Tinea tonsurans in acquired immunodeficiency syndrome]. PMID- 2974552 TI - [Priapism. New pathogenic data and therapeutic consequences]. PMID- 2974553 TI - [Hemodynamic effects of alinidine in the early postoperative phase of cardiac surgery]. PMID- 2974554 TI - [Hyperthyroidism with normal thyrotropin assay disclosing thyrotropic pituitary adenoma]. PMID- 2974555 TI - [Naloxone and naltrexone in the treatment of chronic obstructive bronchopneumopathies]. PMID- 2974556 TI - [Lyme myocarditis without auriculoventricular conduction disorders]. PMID- 2974557 TI - [Severe and uncommon form of murine typhus]. PMID- 2974558 TI - [A new case of Hodgkin's disease in a patient with human immunodeficiency virus seropositivity]. PMID- 2974559 TI - [Pruritus caused by sweat: prevention by ablutions using Vichy Saint-Yorre water]. PMID- 2974560 TI - [Treatment of pulmonary embolism with tissue plasminogen activator]. PMID- 2974561 TI - [Anti-idiotypes in immune glomerulonephritis]. PMID- 2974562 TI - [Current methods of measuring bone density]. PMID- 2974563 TI - [Cerebral edema with hyperammonemia in valpromide poisoning. Manifestation in an adult, of a partial deficit in type I carbamylphosphate synthetase]. AB - A case of cerebral oedema developed during an apparently common attempted suicide with valpromide is reported. The most conspicuous biochemical abnormality was hyperammonaemia. The oedema proved refractory to the standard medical treatment of intracranial hypertension, and decompressive craniectomy was performed with only minor sequelae. The cerebral oedema cum hyperammonaemia syndrome led to the discovery, in this hitherto asymptomatic adult subject, of a 50 per cent deficiency in type a carbamyl phosphate synthetase liver activity. By completing such a deficiency, valproate may produce an extremely serious syndrome resembling the neonatal encephalopathy due to complete enzyme deficiencies in the urea cycle. All valpromide or valproate intoxications probably are cerebral oedemas with hyperammonaemia akin ti Reye's syndrome. All accidents of this type occurring during treatment or poisoning with valproate should be investigated for urea cycle enzyme abnormalities. PMID- 2974564 TI - [Aging of the pancreas. Its implication in malnutrition states in elderly persons]. AB - The influence of ageing on exocrine pancreatic function was investigated in rats and in men. Young rats (3-months old, 150-200 g) and old rats (24-month old, 400 500 g) were killed after fasting overnight. Small pancreatic fragments were removed and kept for stereological analysis both in light and electron microscopy; in addition, levels of tritium-labelled leucine uptake and protein synthesis were measured by quantitative histoautoradiography on isolated acini in vitro. The human study was conducted retrospectively in 27 adults (mean age 36 +/ 1.5 years) and in 28 elderly subjects (mean age 72 +/- 0.6 years) with no clinical or radiological evidence of pancreatitis. Duodenal aspirates were taken over a 90 min period under secretin (0.5 U/kg.h) and cerulein (75 U/kg.h) infusion for comparisons of bicarbonate, lipase, chymotrypsin and amylase concentrations and outputs in the two groups. Elderly people were found to have significant and parallel decreases in bicarbonate, lipase and amylase output as compared to younger subjects (-40 per cent, P less than 0.001). The pancreatic deficiency was confirmed in rats by a significant decrease in zymogen volume density and zymogen diameter and a defect of protein synthesis with significant slowing down (-70 per cent, P less than 0.001) of newly synthesized protein transfer to the Golgian zone of acinar cells. Signs of exocrine parenchyma dystrophy (pancreatitis?) were also observed. PMID- 2974565 TI - [Cystic dilatation of the choledochus. 9 cases]. AB - A retrospective study of 9 cases of extra-hepatic biliary cyst is presented. Prior to admission to our hospital, 5 patients had been operated upon elsewhere. This initial operation proved to be of great importance. In 4 patients initial excision of the cyst was performed resulting in complete regression of symptoms, and no further surgery was required. In 5 patients the cyst was not excised initially and cysto-enterostomy was performed. Recurrence of symptoms and complaints, with major morbidity, occurred in all but one of these patients. After several operations, only final excision of the cyst definitively suppressed the symptoms, but in one patient the cyst could no longer be excised. Cysto enterostomy results in recurrent symptoms and cholangitis and should be avoided. Early excision of extra-hepatic biliary cysts should be performed whenever technically feasible, not only to prevent these complications but also because of the risk of malignant degeneration related to the cyst. PMID- 2974566 TI - [Left ventricular hypertrophy in hypertensive patients. Ultrasonic and physiopathology: therapeutic implications]. AB - Hypertensive left ventricular hypertrophy can easily be diagnosed by echocardiography. It occurs in 30-50 per cent of hypertensive patients and has recently been shown to be a potent and independent predictor of morbidity and mortality, increasing the risk of sudden death, arrhythmia and severe coronary events. The condition usually presents as concentric left ventricular hypertrophy with symmetrically or asymmetrically thickened ventricular walls, but it may also be found with a dilated cavity. Its presence is a definite indication for active medical treatment. Guidelines for the choice of a particular antihypertensive drug are not yet available. Regression of the hypertrophy seems to be a rational goal of the treatment, but the beneficial effects of such a regression on left ventricular relaxation need further evaluation. PMID- 2974567 TI - [Pericardio-peritoneal shunt with the automatic stapler EEA]. AB - Transdiaphragmatic pericardio-peritoneal shunt is an effective answer to the problem of recurrent pericardial effusion. The EEA (end to end anastomosis) stapler is a perfect tool for this technique, being easy to handle and offering the possibility of a pathological examination of the pericardic fragment removed. Five patients underwent this type of surgery with satisfactory results on a 3 to 29 months' follow-up. PMID- 2974568 TI - [Fatal pernicious attack due to chloroquinoresistant Plasmodium falciparum on return from Burkina Faso. 1 case]. PMID- 2974569 TI - [Production of arteriovenous shunts for the free grafts]. PMID- 2974570 TI - [Inflammatory pseudo-tumor of the lung in adults. 1 case]. PMID- 2974571 TI - [Acanthosis nigricans marker of severe insulinoresistance in 2 diabetic patients with android obesity]. PMID- 2974572 TI - [Septic shock caused by Streptococcus milleri after endoscopic sclerosis of esophageal varices]. PMID- 2974573 TI - [Emergence of multiresistant Plasmodium falciparum malaria in Central African Republic]. PMID- 2974574 TI - [Evaluation of a rapid test of measuring antitetanus immunity in the practice of an emergency department]. PMID- 2974575 TI - [The newborn infant of a mother with lupus]. PMID- 2974576 TI - [Early anomalies of CD4 and CD20 lymphocyte cycles in human immunodeficiency virus]. AB - Circadian variations in the number of circulating lymphocytes and their subpopulations have been observed in healthy subjects. These cyclic changes are characterized by a trough at 8:00 a.m. and a peak at midnight. Using multiple peripheral blood samplings, we were able to confirm that this cycle applied to CD4 T-cells (helpers) and to B-cells (CD20). No cycle of CD8 lymphocytes was observed. In a second stage, for greater comfort of the patient the number of samplings was reduced to two: one at 8:00 a.m. (trough) and one at midnight (peak). This method enabled us to calculate the amplitude of lymphocytes cycles in 18 controls and 74 human immunodeficiency virus (HIV) seropositive patients. In asymptomatic HIV carriers the amplitude of CD4 cycles was normal in 6/26 cases and that of B-cell cycles in 2/17 cases. In the group of asymptomatic HIV carriers the mean amplitude of the cycles was much less reduced than in the other two groups. These results incite us to believe that the loss of the CD4 T-cell cycles is an early sign of HIV infection antedating the decrease observed in the number of these cells. PMID- 2974577 TI - [Epidemiology of glomerular diseases in a region in France. Changes as a function of periods and the age of patients]. AB - Between January 1, 1976 and December 31, 1987, a histological diagnosis of primary glomerular disease was made in 420 patients born and living at the time of diagnosis in a region of France with some 400,000 inhabitants. The prevalence of glomerular disease during that span of time was 1 in 1,000. The annual incidence of the disease was evaluated separately for 3 consecutive 4-year periods: period A (1976-79), period B (1980-83) and period C (1984-87). Within each of these 3 periods the number of patients with primary glomerular disease was 138, 148 and 115 respectively, and the figures for annual incidence were 8.6, 9.2 and 7.2 respectively in 100,000. Idiopathic IgA glomerulonephritis was the most common of primary glomerular diseases (34.2 per cent), and its annual incidence remained the same throughout the 3 periods: 2.6, 3.2 and 2.8 in 100,000. The incidence of membranoproliferative glomerulonephritis (7 per cent) decreased from 1980 onward (1.2, 0.5 and 0.3 in 100,000) while that of idiopathic proliferative glomerulonephritis (GNPI) with crescents slightly increased (0.25, 0.7 and 0.5 in 100,000). Acute streptococcal glomerulonephritis virtually disappeared during periods B and C. Lipoid nephrosis was significantly less frequent in period C, whereas secondary membranous glomerulonephritis progressed. There was no significant difference between the 3 periods as regards the incidence of other primary glomerular diseases. All histological types of the primary diseases became more frequent in elderly people. Thus, the overall frequency of primary glomerular diseases decreased slightly during the last 4 years, but it increased in the population aged over 65 and diminished in the younger population. PMID- 2974578 TI - [Recurrent rhabdomyolysis and deficiency of carnitine palmityl transferase. Evidence of the responsibility of a mutant in 2 brothers]. AB - In two brothers who had been presenting since childhood with intermittent muscular attacks and myoglobinuria after sustained efforts, muscle carnitine palmityl transferase (CPT) activity was undetectable by the backward hydroxamate colorimetric method. Such a total deficiency could not easily fit in with the clinical features (normal muscular activity outside sustained efforts), with the normal results obtained at electromyography and with the moderate fatty overload detected at muscle biopsy. In order to elucidate these apparently discordant findings, another CPT measurement method, the forward optimized isotopic technique, was used. With this method, the catalytic activity of CPT can be measured, and enzyme inhibition by its substrate and the product of the reaction (palmityl CoA and palmitylcarnitine respectively) can be studied. The results showed that the catalytic activity of CPT was preserved in both patients, but the enzyme was abnormally sensitive to inhibitors. These 2 cases tend to demonstrate the existence of a mutant carnitine palmityl transferase. PMID- 2974579 TI - [Initial treatment of infiltrating tumors of the bladder. Combined transurethral resection and systemic chemotherapy]. AB - From September 1983 to September 1986, 20 patients (mean age 65 years) with a muscle-infiltrating tumour of the bladder would normally have been treated by total cystectomy. Instead, they were staged by intravenous urography, pelvic and abdominal computerized tomography, physical examination under general anaesthesia and deep transurethral resection, then given neoadjuvant chemotherapy consisting of cisplatinum and 5-fluorouracil, six courses at intervals of 28 days. Results were evaluated after the 3rd and 6th courses by computerized tomography, intravenous pyelography and transurethral resection. Nine patients had a clinical complete response (6 pT2, 2 pT3, 1 pT4). The median follow-up in january, 1988 was 30 months (range 17-52 months). This protocol was objectively active and well tolerated, even by elderly subjects. Two problems remain concerning patients with complete response: the respective roles of chemotherapy and transurethral resection in the outcome, and the prevention of recurrence (5/9 complete response patients). PMID- 2974580 TI - [Thoracic x-ray computed tomography in pneumology]. AB - Computerized tomography (CT) of the chest is now widely used in pneumological practice. It has completely superseded conventional tomography and reduced the number of bronchographies and pulmonary angiographies. CT is superior to conventional examinations in detecting lymph node enlargements, solid tumours, granulomas, fat deposits, calcifications, pulmonary necrosis and pleural plaques and in demonstrating tumoral limits. In the lung itself, the information provided by CT is of subsegmental accuracy. The new method is capable of diagnosing bronchiectasis and emphysema and evaluating their extension; it also gives a good visualization of chronic interstitial lung lesions. PMID- 2974581 TI - [Pleural localization of multiresistant non-typhoid Salmonella]. PMID- 2974582 TI - [Treatment of chronic delta hepatitis with recombinant alpha 2 interferon in 8 patients with anti-human immunodeficiency virus antibodies]. PMID- 2974583 TI - [Effect of a growth hormone inhibitor on the course of Duchenne's myopathy]. PMID- 2974584 TI - [Hodgkin's disease in 2 human immunodeficiency virus seropositive women]. PMID- 2974585 TI - [Diabetogenic effects of estrogens and progestins in therapy]. PMID- 2974586 TI - [Epidemiology of chronic renal insufficiency treated by dialysis in a region in France. Changes in a 12-year period]. AB - Between January 1976 and December 1987, 278 patients (172 men, 106 women) aged over 15 and living in a region of some 400,000 inhabitants were treated with dialysis for end-stage chronic renal failure (CRF). The annual incidence of new cases treated was evaluated separately for 3 consecutive 4-year periods: 1976-79 (period A), 1980-83 (period B) and 1984-87 (period C). The incidence rose from 4.6-4.9 in periods A and B to 7.8/100,000 in period C, i.e. a 38 per cent progression. At the start of dialysis the mean age of women did not significantly vary throughout the 3 periods, whereas it increased significantly in men during period C. There was a significant increase of primary glomerulonephritis (GN) throughout the 3 periods: 1.25, 1.70 and 2.35/100,000 respectively; this disease accounted for 27.3, 34.6 and 30.6 per cent of all causes of CRF. Similarly, the incidence of secondary nephropathy (diabetes, amyloidosis) treated with dialysis increased from 0.6 (A) to 1.0 (B) and 1.9/100,000 (C) (P less than 0.05 with period A). The same happened with polycystic kidney: 0.25, 0.65 and 0.93/100,000. During period C, when 95 per cent of primary GN were diagnosed histologically, igAGN (13 per cent), diabetes (13 per cent) and polycystic kidney (12 per cent) were the main causes of CRF. During the whole 12-year period under study, when 80 per cent of primary GN were diagnosed, the prevalence of causes of CRF varied according to the patients' age. In patients under 65 at the start of dialysis primary GN were the principal causes of CRF (36.8 per cent, including 15.5 per cent of IgAGN), followed by secondary nephropathies (20.3 per cent, including 10.7 per cent of diabetes), chronic interstitial nephropathies (18.7 per cent, including 9.6 per cent of reflux nephropathy), hereditary nephropathies (14.4 per cent, including 10.7 per cent of cystic kidney) and vascular nephropathies (7 per cent). Above the age of 65, the principal causes of CRF were chronic interstitial nephropathies (27.7 per cent, including 8.9 per cent of drug-induced nephropathy), followed by secondary nephropathies (20 per cent, including 6.7 per cent of amyloidosis and 11.1 per cent of diabetes), vascular nephropathies (20 per cent), primary GN (12.2 per cent), and hereditary nephropathies (12.2 per cent) including 11.1 per cent of polycystic kidney).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2974587 TI - [Animal bites. Epidemiology and infection risks]. AB - A review of 5,116 cases of animal bites (587 of which were studied prospectively) has shed some light on their epidemiological aspects and on the risk of infection they carry. It has also led to a more objective assessment of the real effect of the therapeutic and prophylactic measures usually applied in such cases. The most frequent wounds are those of the hands and face, the former rising an infectious problem, the latter a predominantly cosmetic problem. The overall risk of infection is 30 per cent, but it is increased, notably as regards pasteurellosis, in the case of cat bite. Precise and simple rules concerning the prevention of this risk cannot easily be given, but it seems that the systematic antibiotic treatment initially prescribed has not clearly proved effective. Similarly, early sutures do not significantly increase the risk of infection. PMID- 2974588 TI - [Erythermalgia, rare acrosyndrome. 13 cases]. AB - The diagnosis of erythermalgia, initially made in 27 patients between 1980 and 1986, was re-evaluated on the basis of 7 criteria. Three were major criteria: paroxysmal attacks, burning pain in the extremities and redness of the territory concerned during the attacks. The 4 minor criteria were: typical precipitating factors (exposure to heat, effort), typical relieving factors (exposure to cold, rest), elevated local temperature during the attacks and response of symptoms to acetylsalicylic acid. The diagnosis was deemed to be correct when the 3 major criteria and at least 2 of the minor criteria were present. Thirteen patients (8 women, 5 men) fulfilled these conditions. Nine of them had primary erythermalgia and in 4 patients the condition was consecutive to a myeloproliferative syndrome (thrombocytopenia in 2 cases, Vaquez' disease in 2 cases). These two forms differed on several points. Patients with secondary erythermalgia were older, some had unilateral disorders, and their symptoms were less intense; the syndrome always followed a favourable course and disappeared when the causative disease was cured; in 3 out of 4 cases erythermalgia disclosed a myeloproliferative syndrome. Patients with primary erythermalgia were younger, the syndrome was of longer duration and the symptoms always bilateral and sometimes severe; those who responded to acetylsalicylic acid had a favourable prognosis, but treatment was difficult in the others. Capillaroscopy is of little help to diagnose this syndrome; this is done on clinical grounds only and it is easy when the criteria, as defined in this study, are present. In every case, blood examination with platelet count and erythrocyte sedimentation rate is advisable. PMID- 2974589 TI - [Blood histamine levels in patients infected with the human immunodeficiency virus]. AB - Serum histamine levels are significantly lowered in patients with human immunodeficiency virus (HIV) infection. Treatment with zidovudine has no effect on histaminaemia, but HIV-positive patients with thrombocytopenic purpura who underwent splenectomy are known to have had a rise in serum histamine levels after the operation. This parameter might be used as prognostic factor in the evaluation of patients with HIV infection. PMID- 2974591 TI - [Retrograde section of uterine septa under ultrasonic control]. AB - A new technique for the treatment of uterine septa is described. The septum is divided by the transcervical route, using 4 mm endoscopic scissors. Simultaneously, real-time ultrasonography confirms the diagnosis of septate uterus and ensures the safety of this blind procedure. Ultrasonic guidance also enables the septum to be divided along the medial antero-posterior plane and the distance between the upper limit of the section and the fundus uteri to be measured. The operation is rapid, carries a limited risk, leaves no uterine or abdominal scar and requires no special equipment. The elective indication for section of uterine septa is habitual abortion. This convenient method can also be used in women who have never been pregnant during coelioscopy for sterility or section of a vaginal septum. PMID- 2974590 TI - [Basal-cell nevomatosis associated with multifocal fetal rhabdomyoma. A case]. AB - Nevoid basal-cell carcinoma is a hereditary syndrome. Its major features are a multiple basal-cell carcinoma which appears early in childhood, skeletal and genital abnormalities and ectopic calcifications. It may be associated with malignant schwannoma, medulloblastoma and lymphoma. We report one case of nevoid basal-cell carcinoma syndrome associated with foetal rhabdomyoma, thyroid gland polyadenoma and benign schwannoma. The first case of foetal rhabdomyoma associated with this syndrome was described in 1976. PMID- 2974592 TI - Activity of luxabendazole against liver flukes, gastrointestinal roundworms, and lungworms in naturally infected sheep. AB - The anthelmintic potential of luxabendazole was investigated in sheep harboring mixed naturally acquired helminth infections. Results were assessed by comparing worm counts of the treated groups (seven animals each) on days 7-8 posttreatment with those of the nontreated control group, except for protostrongylid lungworms, for which the changes in pre- and posttreatment group mean larval counts/g feces were assessed for intensity effect. A single oral treatment at doses of 10.0 or 12.5 mg/kg body wt removed 97.6% of the adult Fasciola hepatica and 63.2%-83.8% of the Dicrocoelium dendriticum. Luxabendazole at 7.5, 10.0, and 12.5 mg/kg proved 100% effective in removing adult worms of the genera Haemonchus, Ostertagia, Trichostrongylus, Cooperia and Nematodirus as well as tissue associated larval stages of gastrointestinal nematodes of the abomasal mucosa. The drug showed an intensity effect of 79.7%-87.6% against Strongyloides papillosus. Luxabendazole removed all Dictyocaulus filaria and reduced the fecal excretion of larvae of protostrongylid species (Protostrongylus rufescens, Neostrongylus linearis, Cystocaulus ocreatus, Muellerius capillaris) by 97.8% 99.6%. The efficacy of luxabendazole compared favorably with that of Diplin Kombi (oxyclozanide and levamisole), which was used as a reference drug. PMID- 2974593 TI - The thrombolysis in myocardial infarction (TIMI) trial phase I and phase IIA pilot results. Implications for nursing. PMID- 2974594 TI - Role of atrial natriuretic factor in regulation of blood pressure in normotensive rats having reduced renal mass. AB - Experiments were carried out in normotensive, saline-drinking, 60% reduced renal mass rats to determine the effect of an in vivo blockade of endogenous atrial natriuretic factor (ANF) on blood pressure. We used a 60% reduction in renal mass because blood pressure in these normotensive animals is extremely sensitive to any slight further reduction of renal excretory function. Six weeks following the reduction of renal mass and documentation of normotension, rats were injected intraperitoneally twice daily for 12 days with ANF antibody prepared against the C-terminal heptapeptide of AP III conjugated to bovine thyroglobulin. Control rats similarly prepared, received normal rabbit serum (NRS). Blood pressure progressively increased in rats receiving the antibody, and its withdrawal returned blood pressure to control levels within 4-5 days. Serum from either normal rabbits or rabbits immunized with bovine thyroglobulin or peptides unrelated to ANF had no effect on blood pressure in the control animals. These experiments show that in the normotensive saline-drinking rat with reduced renal mass, an antibody to AP III raises blood pressure. This suggests that ANF here is acting to prevent the rise in blood pressure. PMID- 2974595 TI - Catabolism of 5-fluoro-2'-deoxyuridine by isolated rat intestinal epithelial cells. AB - The kinetics of conversion of 5-fluoro-2'-deoxyuridine (FdUrd) to 5-fluorouracil (FUra) by isolated rat intestinal epithelial cells was investigated. Also, the effects of potential inhibitors of this reaction, which is catalyzed by uridine phosphorylase and thymidine phosphorylase, were determined. A 2.5% suspension of isolated cells was incubated with FdUrd or FUra, and at specific times cells were lysed with perchloric acid and fluoropyrimidines were determined by high performance liquid chromatography. During a 25-min incubation with either FdUrd or FUra, the amount of drug in the incubation system (total volume 0.8 ml) fell by less than 5%. However, in the presence of FdUrd, the amount of FUra increased linearly over 25 min. The apparent Vmax and Km for FUra formation were 17-27 nmole/mg DNA/min and 1.6-2.5 mM, respectively. With each nucleoside phosphorylase inhibitor, the apparent Km increased but Vmax was unaffected. The apparent Ki values were as follows (in mM): 5-nitrouracil (an inhibitor of both uridine phosphorylase and thymidine phosphorylase), 0.12; 4-thiothymine (a uridine phosphorylase-selective inhibitor), 1.52; and 6-benzyl-2-thiouracil (a thymidine phosphorylase-selective inhibitor), 0.73. It was concluded that intestinal epithelial cells are capable of degrading FdUrd to FUra and that the cells possess both uridine phosphorylase and thymidine phosphorylase activity. PMID- 2974596 TI - Promotion of pulmonary carcinogenesis by plutonium particle aggregation following inhalation of 239PuO2. AB - Promotion of lung tumor formation from inhaled 239PuO2 in rats may be associated with aggregation of plutonium particles near bronchioles. The relationship of plutonium particle aggregation in the lung and the development of lung tumors after inhalation of 239PuO2 was studied in 664 life span rats with mean lung doses ranging from 0.35 to 20 Gy. Plutonium particle concentration and aggregation were determined from autoradiographic sections of the left lung lobe. The increase in particles/cm2 and mean number of particles per aggregate up to 20 Gy were directly proportional to lung dose. Aggregates with greater than 25 particles increased linearly with dose from 0.2% at 1.4 Gy to 8.2% at 20 Gy, in a pattern similar to increasing severity of pulmonary fibrosis and incidence of lung tumors. Lung tumor incidence increased from about 6% at 1.4 Gy to 83% at 8 Gy; no further increase in lung tumors was seen at doses greater than 8 Gy. Maximum lung tumor incidence at 8 Gy corresponded to a particle concentration of 130/cm2 and four particles/aggregate with 4% of aggregates having greater than 25 particles. Aggregation of inhaled plutonium particles in clusters of greater than 25 particles resulted in daily doses of only a few centigray to focal tissue regions containing clustered particles, yet these doses appeared sufficient to cause pulmonary fibrosis and promotion of pulmonary carcinogenesis. PMID- 2974597 TI - Legal case briefs for nurses. MI.: injury in route to job: compensation; GA.: side rail use: expert testimony. PMID- 2974598 TI - [Echographic aspect of 2 rare soft-tissue neoplasms: chordoid sarcoma and neurotekeoma]. PMID- 2974599 TI - Social validation of decelerative (punishment) procedures by special educators of severely handicapped students. AB - Ratings on the acceptability, intrusiveness, restrictiveness, and efficacy of 18 commonly used interventions for decelerating behaviors were obtained from 58 special educators of students with severe handicaps. Results indicated that consensually high and low mean ratings were obtained for restrictiveness, intrusiveness, and acceptability. Ratings of efficacy, however, were generally neither consensually high nor low, and demonstrated lower variability across the 18 interventions, and lower reliability across raters than did ratings on the other three dimensions. Additionally, ratings of restrictiveness were negatively correlated with ratings of acceptability, ratings of intrusiveness and restrictiveness were positively correlated, and ratings of acceptability and efficacy were positively correlated. We interpreted these results as suggesting that: (a) special educators may not be certain of the efficacy of interventions they use, despite previously reported use in classrooms, and (b) the constructs of restrictiveness and intrusiveness may not have discriminant validity at present. Implications for ongoing training for special educators were discussed. PMID- 2974601 TI - An attempt to improve auditory short-term memory in Down's syndrome individuals through reducing distractions. AB - Down's syndrome (DS) individuals, relative to nonretarded individuals, have greater difficulty remembering brief sequences of verbal information presented auditorily. Previous research suggests at least two possible attentional explanations of their difficulty: They are especially susceptible to both auditory distraction and off-task glancing during laboratory tasks. DS, non-DS mentally retarded and nonretarded persons listened to, looked at, and attempted to remember sequences of digits. Although the three groups did not differ in their recall of visually-presented stimuli, DS subjects showed significantly poorer recall of auditorially-presented stimuli than the other two groups (which did not differ). Furthermore, the poor auditory memory of DS subjects did not improve under testing conditions designed to minimize auditory and visual distractions. It was suggested that poor auditory short-term memory for verbal information is tied more closely to Down's syndrome than to low intelligence and does not seem to be caused by a special susceptibility of Down's syndrome individuals to attentional distractors. PMID- 2974600 TI - Stereotypic behavior of mentally retarded adults adjunctive to a positive reinforcement schedule. AB - Stereotypic behavior is one of the more common disturbed behaviors displayed by people who are developmentally disabled. This study evaluated the indirect effects on stereotypic frequency when the value of a concurrent fixed-interval reinforcement schedule for adaptive behavior was varied. Three profoundly mentally retarded adults performed a simple adaptive task reinforced under a fixed-interval schedule. The reinforcement schedule value was varied from fixed interval 15 to 90, and 180 seconds after schedule control under each condition was demonstrated. The dependent measure was the frequency of stereotypic behavior. Stereotypic behavior increased in direct relation to the interval length. The theoretical and practical implications of treating stereotypies as an adjunctive behavior partially controlled by the reinforcement frequency for adaptive behaviors are discussed. PMID- 2974602 TI - A graduated guidance procedure for teaching self-dressing skills to multihandicapped children. AB - The effectiveness of a graduated guidance procedure for increasing independence in dressing was examined in a multiple baseline analysis across behaviors. Two multihandicapped children were trained to dress in socks, pants, and shirt. In training sessions, subjects completed the entire sequence of steps involved in putting on the training garment on each trial. Trainer assistance was provided as necessary, but was faded systematically according to a hierarchy of intrusiveness. Positive reinforcement was delivered contingent on dressing with increased independence. Assessment of independent responses followed each training session. Dependent measures were derived from a task analysis of each dressing behavior. Results showed that both children learned dressing skills, although rates of acquisition varied considerably. Generalization of treatment effects to similar garments was observed; maintenance of skills was evident at follow-up probes conducted at 36 weeks for one child and 18 weeks for the other. PMID- 2974603 TI - [Clinical comments on testicular tumors seen at a urology clinic]. PMID- 2974604 TI - [Acute pancreatitis due to an apparent nonbiliary cause]. PMID- 2974606 TI - [A complex arm wound resolved by an epiploon autograft-- microsurgical technics]. PMID- 2974605 TI - [Laparostomy in the treatment of acute pancreatitis]. PMID- 2974607 TI - [Congenital cystic dilatation of the biliary tract]. PMID- 2974608 TI - [Intestinal occlusion in submucosal sigmoid lipoma]. PMID- 2974609 TI - [Persistent oculocardiac reflex following a retrobulbar hematoma]. PMID- 2974610 TI - [Fulminant form of intra-anesthetic malignant hyperthermia]. PMID- 2974612 TI - Family support for the disabled. PMID- 2974611 TI - [Peridural anesthesia with ketamine]. PMID- 2974613 TI - [Dental management of severely handicapped children. (Report 1) Evaluation of using caries activity tests]. PMID- 2974614 TI - [Caries-inhibitory effect of mutastein in animal experiments]. PMID- 2974615 TI - [Effect of TG syrup on cariogenic properties of Streptococcus mutans]. PMID- 2974616 TI - [Dental treatment of handicapped patients during a 5-year period (1980-1984) under general anesthesia and ambulatory clinic, at Tokyo Infant Recuperation Hospital]. PMID- 2974618 TI - [Gynecologic endoscopy]. PMID- 2974617 TI - [Dental treatment for handicapped children under general anesthesia at the Pedodontic Clinic in Osaka University Dental Hospital]. PMID- 2974619 TI - [Coronary angioplasty in 1988]. PMID- 2974620 TI - Increasing the functional residual capacity may reverse obstructive sleep apnea. AB - We describe the reversal of obstructive sleep apnea with a 0.5 L increase in the functional residual capacity (FRC) in a patient with sleep apnea syndrome. The patient had been treated with medroxyprogesterone acetate for 8 months. The increase in FRC was obtained by applying a constant negative extrathoracic pressure (NEP) with a poncho-type respirator. With pulmonary inflation, there was a dramatic decrease in the apnea index and the percent apnea time, and an improvement in sleep architecture. At all sleep stages, the desaturation duration was shorter with NEP. The exact mechanisms by which pulmonary expansion improved sleep apnea in this patient remain unclear; lung volume dependence of upper airway patency and the improvements in apnea-induced desaturation may be contributing factors. Our observation illustrates that lung volumes may be an important factor in the pathophysiology of obstructive sleep apnea, especially in the apnea onset and in the apneic-induced desaturation. PMID- 2974621 TI - [Current therapeutic approach to adnexal cysts]. AB - From a 5-year study of 481 ovarian tumors, the authors discuss their therapeutic management and try to answer two basic questions: what is the proportion of cystic tumors, that actually are carcinomas? To what extent the tapping of a cancerous "cyst" may cause the spreading of the tumor? PMID- 2974622 TI - Atrial content and plasma levels of atrial natriuretic peptides in rats with chronic renal failure. AB - The possible role of atrial natriuretic peptides (ANP) for the adaptive changes in renal Na excretion during chronic renal failure was studied in 5/6 nephrectomized (NX) rats maintained on a normal (100 mmol/kg) and a high (800 mmol/kg) Na diet. Atrial content of natriuretic substances was determined by bioassay and plasma ANP by radioimmunoassay. Nephrectomized rats showed a twofold increase in plasma ANP irrespective of their Na intake. Atrial ANP content was increased by high Na diet but unchanged by NX. Nephrectomized rats maintained on high Na diet showed partial depletion of atrial ANP stores. There were no significant changes in the volume fraction of atrial granules determined. The results suggest that ANP is involved in the regulation of renal Na excretion during chronic renal failure and acute Na loading; other mechanisms are probably involved in the adaption to chronic Na loading. PMID- 2974623 TI - Effect of load-cycling on bond between composite fillings and dentin established by Gluma and various resins. AB - Cylindrical dentin cavities, pretreated with Gluma and various resins, were filled with Silux, P-30, or Concise. Marginal integrity was in most instances obtained when experimental resins containing propionic aldehyde were used. The bond between composite and dentin established in this way was tested by load cycling. Cervical located cavities in extracted human teeth, approximately one half of the margins in dentin, were etched and treated as above. Fillings made of Silux, P-30, or Concise exhibited margins without stain upon axial loading in most cases. The frequency of staining increased when Dycal was used as cavity liner, but were absent in all cases when Concise was used as filling material. The content of propionic aldehyde in the intermediary resin layer may reduce the oxygen tension at the interface and thereby increase the rate of the polymerization at this location. PMID- 2974624 TI - Interactions of protein S with membranes. PMID- 2974625 TI - The use of single photon emission computed tomography (SPECT) in the diagnosis of low-back pain in young patients. AB - Single photon emission computed tomography (SPECT) is a method of scintigraphy that provides sectional and multi-planar imaging that should improve the sensitivity for diagnosis in remote areas of the spine. To test this hypothesis, the authors reviewed 15 patients with low-back pain and compared the results of the radiographs, planar bone scans (PBS), and SPECT. Four of 15 had normal examinations with all three methods of imaging. The rest had a positive test in at least one of the three methods. Single photon emission computed tomography was positive in eleven, PBS in six, and the radiograph in three of the 11 patients. It appears that the SPECT was the most sensitive method of imaging and greatly enhanced our diagnostic acumen for stress fractures or stress reactions of the spine. PMID- 2974626 TI - Clinical and radiological evaluation of lumbosacral motion below fusion levels in idiopathic scoliosis. AB - The purpose of this study was to evaluate the unfused segments of the lumbar spine in patients who had Harrington instrumentation and fusion for idiopathic scoliosis. Forty-eight patients, with an average follow-up of 11 years, were evaluated. The translational motion in the unfused segments below the instrumented levels was measured, using lateral flexion and extension radiographs of the lumbar spine. This motion was compared with values obtained from an earlier study of asymptomatic nonscoliotic individuals. The amount of disc space narrowing, retrolisthesis, length and level of the fusion, and the presence of traction spurs also were recorded. The incidence of low-back pain was highest in those patients fused to L4 (62%). Individuals instrumented and fused to L3 or L4 had significantly more translational motion in the adjacent lower interspace when compared with the control group (P = 0.05 and P = 0.001, respectively). Increased translational motion correlated with the presence of low-back pain in patients fused to L4. Retrolisthesis occurred in 81% of patients instrumented to L4, in 40% of those fused to L3, and was not found in patients fused to high levels. Its presence was strongly associated with low-back pain. There was no relationship between low-back pain and traction spurs, length of the fusion mass, lumbar lordosis, or width of the disc space in the unfused lower levels. The authors conclude that retrolisthesis and increased translational motion are important factors in determining low-back pain following surgery for idiopathic scoliosis. Instrumentation to L4 should be avoided if possible. PMID- 2974627 TI - Treatment, follow-up, or discharge. PMID- 2974629 TI - Lumbar degenerative kyphosis. Clinical, radiological and epidemiological studies. AB - We suggest that lumbar degenerative kyphosis be included as one of the abnormal sagittal curvatures in which a kyphosis or a marked loss of lordosis is seen in the lumbar spine, caused by degenerative changes in middle-aged and elderly. One hundred and five consecutive patients were investigated, most of whom complained of low-back pain, often with a long history. They all walked in a forward bending posture, either all the time or only when exhausted. In roentgenograms, most cases showed a marked loss of the sacral inclination, as well as multiple disc narrowing and/or vertebral wedging in the lumbar region. These subjects showed a definite weakness of the lumbar extensors compared to the flexors, and therefore a reversed ratio of extensors/flexors muscle power compared with normal controls and other types of spinal curvatures. Weakness of the lumbar extensors was clearly shown by isokinetic measurement and a marked atrophy of these muscles with fatty infiltration was demonstrated by CT scanning. PMID- 2974628 TI - Use of oral colchicine in low-back pain. PMID- 2974630 TI - Neurogenic claudication and root claudication treated with calcitonin. A double blind trial. AB - Forty-two patients with either neurogenic claudication or unilateral root claudication were analyzed in a double-blind comparison of salmon calcitonin (SCT) and placebo, receiving either 100 IU SCT or 1 ml saline four times a week for 8 weeks. Five of 20 SCT and one of 22 placebo patients were classified as responders. There was no statistically significant difference between the treatment groups in the proportion of responders. Seven of eighteen of the placebo group who later received salmon calcitonin improved their walking distance. The authors have not established that this is an organic response. PMID- 2974631 TI - Low-back pain and respiratory function. PMID- 2974632 TI - 1988 Volvo award in clinical sciences. Facet joint injection in low-back pain. A prospective statistical study. AB - From January 1980 through December 1984, 454 patients were evaluated with facet joint injections. All had the chief complaint of low-back pain, normal neurologic examinations and no root tension signs. Three hundred and ninety completed the protocol, which included a lumbar motion pain assessment before and after facet injection. A total of 127 variables were studied. There were 229 males and 161 females with a median age of 38. Facet joint arthrograms were performed prior to intra-articular injection of local anesthetic and cortisone. Initial mean pain relief was only 29%. Variables correlating significantly (P less than 0.05) with more postinjection pain relief were older age, prior history of low-back pain, normal gait, maximum pain on extension following forward flexion in the standing position, and the absence of leg pain, muscle spasm and aggravation of pain on Valsalva. Greatest pain relief immediately after injection was seen with lumbar extension and rotation, motions reported to stress the facet joints or aggravate pain of facet joint origin. Patients with more pain on lumbar extension and rotation as a group, however, did not get more pain relief. From this study we were not able to identify clinical facet joint syndromes or predict patients responding better to this procedure. The facet joints were not commonly the single or primary source for low-back pain in the great majority (greater than 90%) of patients studied. PMID- 2974633 TI - Progressive isoinertial lifting evaluation. II. A comparison with isokinetic lifting in a disabled chronic low-back pain industrial population. AB - The Progressive Isoinertial Lifting Evaluation (PILE), as described in Part I of this series of articles, is a simplified test combining psychophysical and isoinertial protocols to provide an unconstrained lifting assessment. In the second part of this study, 100 chronically disabled low-back pain patients (57 men and 43 women) were studied at two points: 1) at initial evaluation, when referred for possible entry into a comprehensive Functional Restoration treatment program; and 2) at the conclusion of the treatment (an average 7 weeks later). Results of simultaneous lumbar PILE and Cybex Liftask (Lumex, Ronkonkoma, NY) tests are presented, showing that patients may frequently double or triple initial lifting capacity after undergoing the functional restoration training program, achieving lifting levels at or above normal for incumbent industrial workers. Overall, results demonstrate that the PILE test can be an effective baseline screening test for lifting capacity under certain circumstances. Although several drawbacks affecting the PILE as an isolated test are discussed, its usefulness as part of a battery of physical capacity tests making up a quantitative functional evaluation is clearly demonstrated. Finally, the potential use of PILE as a safe, inexpensive, simple, and relevant screening test for frequent lifting capacity in worker selection is discussed. PMID- 2974634 TI - Intracoronary thrombolysis and coronary angioplasty for evolving myocardial infarction. AB - At present experience in South Africa with coronary reperfusion during the acute phase of myocardial infarction is limited. Acute reperfusion of an infarct related coronary artery was attempted in 55 patients using intracoronary thrombolytic therapy and percutaneous transluminal coronary angioplasty (PTCA). Intracoronary streptokinase infusion had reopened 25 to 40 totally occluded lesions but usually a residual severe stenosis remained. PTCA was subsequently attempted in 36 severely stenotic coronary arteries and 19 totally occluded coronary arteries. PTCA was successful in 48 of the 55 cases (87%). After 1 week vessel patency was present in 36 of the 39 patients who had early coronary artery re-assessment. Late restenosis occurred in 4 out of 14 cases. There were no procedure-related deaths. Normal global as well as regional left ventricular function was present in 15 cases after 1 week. Emergency PTCA alone or in combination with intracoronary thrombolytic therapy is efficacious in achieving coronary reperfusion. On follow-up, left ventricular function appeared to be well preserved in 27% of patients with patent infarct-related arteries. PMID- 2974635 TI - [Nonsurgical recanalization of occluded femoral and popliteal arteries by a combination of the Dotter technic, angioscopy and atherectomy]. PMID- 2974636 TI - [Rotation angioplasty--a new technic for reopening chronic arterial occlusions]. PMID- 2974637 TI - [Assessment of thrombocyte deposits and thrombocyte survival following catheter dilatation of the lower extremities using 111-indium marked thrombocytes]. PMID- 2974638 TI - [Vascular surgery aspects of indications for percutaneous transluminal angioplasty of the popliteal area]. PMID- 2974639 TI - [Significance of percutaneous transluminal angioplasty in the treatment of renovascular hypertension from the vascular surgery viewpoint]. PMID- 2974640 TI - [Renal lesions in patients infected with human immunodeficiency virus]. PMID- 2974641 TI - [Methodological problems and various results of the study of the incidence and structure of diseases of the urinary system among adult population (data from outpatient clinics of Moscow, Celinograd and the Celinograd district of the Kazakh SSR)]. AB - A 4-stage system of mass screening of population for the detection of renal diseases tested in one of the outpatient clinic zones in Moscow (2013 residents), Thelinograd (7091 residents), and Tselinograd Region, Kazakh SSR (880 residents) was presented. The incidence rates of renal diseases per 1000 examined population in Moscow, Tselinograd and Thelinograd Region, Kazakh SSR, were 12.4 +/- 2.5, 10.4 +/- 1.2, 13.6 +/- 3.9, respectively. PMID- 2974642 TI - [Current approaches to immunological diagnosis of autoimmune and immune complex diseases]. PMID- 2974643 TI - [Detection of pathology of the urinary system in students during preventive mass screening]. AB - Screening tests including the questionnaire method, urine analysis and radiorenography were used during mass screening of 1330 students. Comparison of the results of the first two methods showed a good informative value of the questionnaire method for the detection of pathology of the kidneys and urinary tracts. Radiorenography performed in 106 students with the detected nephrological symptom complex (risk group) revealed some changes practically in every other "risk group" student. PMID- 2974644 TI - [Myocardial aspects of nephrogenic hypertension and its treatment with captopril]. PMID- 2974645 TI - [Acetylation phenotype in patients with periodic disease]. PMID- 2974646 TI - [Fatty acids of the omega-3 line in the erythrocytes of Chukchi with myocardial hypertrophy]. AB - A study was made of the fatty acid composition of erythrocytes in the Chukchi aged 30 to 59 with myocardial hypertrophy and in controls. In erythrocytes of the Chukchi the percentage of polyunsaturated fatty acids of omega-3 series including eicosapentaenoic acid was increased (C 20:5), and the percentage of linoleic acid and arachidonic acid was decreased (C 18:2 and C 20:4, respectively) as compared to similar indices of erythrocytes in Muscovites. Myocardial hypertrophy was diagnosed in the Chukchi in whom the percentage C 20:5 in erythrocytes was higher and exceeded that in PL. It was assumed that changes in the fatty acid composition of cell membranes played certain part in disorder of their function and the development of myocardial hypertrophy in the Chukchi. PMID- 2974647 TI - Recent studies of the synthetic selective inhibitors; with special reference to non-plasmin fibrinolytic enzyme, plasmin and plasma-kallikrein. PMID- 2974648 TI - Plasminogen activation by tissue plasminogen activator in the presence of platelets. AB - Platelets were found to provide a surface for activation of plasminogen by the tissue-type plasminogen activator (t-PA) at an optimum concentration and to potentiate the generation of plasmin by the amidolytic method, fibrin lysis time and fibrin plate method. The effect of platelets on amidolytic activity on S-2251 was due to the potentiating effect of plasminogen activation by t-PA, because it was observed only in the presence of both plasminogen and t-PA. Plasmin generation was also evidenced in the SDS-PAGE profile of the supernatant from a mixture containing t-PA and plasminogen with platelets. These findings suggests that the potentiating activity of platelets on plasminogen activation by t-PA in circulation is one of the causes of fibrinogenolysis during fibrinolytic therapy with a high dose of t-PA. Platelets from patients with various diseases showed different potentiating activity on plasminogen activation by t-PA. The assay of this ability of platelets may be a new tool for evaluating their role in the blood fibrinolytic process. PMID- 2974649 TI - [A follow-up study of children with birth weights below 1,500g. A multicenter study of children born in Health region 2 during the period October 1, 1981 September 30, 1983]. PMID- 2974651 TI - [Hospitalized patients with backpain examined by means of myelography]. PMID- 2974650 TI - [Chronic backpain with and without organic findings. Discriminating factors]. PMID- 2974652 TI - [Recanalization of chronic occlusion of coronary arteries with transluminal balloon technic]. PMID- 2974653 TI - Pulmonary platelet sequestration is increased following monocrotaline pyrrole treatment of rats. AB - 111In-labeled platelets were used to study the localization and survival of circulating platelets at various times after a single, intravenous administration of 3.5 mg/kg monocrotaline pyrrole (MCTP) to rats. Lung injury, assessed from elevated lung weight, lavage fluid total protein and albumin concentrations, and lactate dehydrogenase activity, was evident at Days 8 and 14. In addition, right ventricular hypertrophy was manifested by 14 days after MCTP administration. Pulmonary sequestration of 111In-labeled platelets was also elevated by Days 8 and 14, while circulating blood platelet number remained unchanged. Concomitantly, the hemoglobin concentration and total hemoglobin content of the lung homogenate supernatant in MCTP-treated rats on these days was decreased when compared to those in controls. A decrease in splenic platelet sequestration on Day 14 was accompanied by an increase in the combined radioactivity of the heart and kidneys. Platelet half-life and mean life span were increased only on Day 14. A higher dose of MCTP (35 mg/kg) caused moderate lung injury at 6 hr. However, this treatment did not result in increased platelet sequestration in the lungs, although a trend was observed. Data from this study support the hypothesis that platelets are involved in the development of the pulmonary hypertensive response following MCTP-induced lung injury. PMID- 2974654 TI - Effect of aspartame on seizures in various models of experimental epilepsy. AB - We investigated in rats whether aspartame intake affected the susceptibility to seizures induced chemically (metrazol, quinolinic acid) or electrically (electroshock). Aspartame (0.75-1.0 g/kg), given orally as a single bolus to 16 hr fasted animals 60 min before metrazol, significantly increased the number of animals showing clonic-tonic seizures. At 1.0 g/kg the ED50 for clonic-tonic convulsions was lowered by 23%. A similar increase in seizure susceptibility was observed with 0.25-0.5 g/kg of the aspartame's metabolite phenylalanine. When aspartame was administered to fasted rats in three divided doses (0.33 g/kg) over 120 min or to fed animals after a meal, or overnight with the diet, no significant changes in the incidence of animals showing seizures was observed. One gram per kilogram aspartame and 0.5 g/kg phenylalanine did not modify the CC50 (mA) for tonic hindlimb extension induced by electroshock and the electroencephalographic seizures caused by intrahippocampal injection of 120 nmol quinolinic acid. Plasma and brain levels of phenylalanine and tyrosine significantly raised after both 1 g/kg aspartame as a single bolus (plasma: Phe 285%, Tyr 288%; brain: Phe 146%, Tyr 192%; above controls) or in three divided doses (plasma: Phe 207%, Tyr 315%; brain Phe 103%, Tyr 211%; above controls) and 0.5 g/kg phenylalanine (plasma: Phe 339%, Tyr 410%; brain: Phe 219%, Tyr 192%; above controls), but the ratio Phe/Tyr was not modified. Our data indicate that aspartame cannot be regarded as a general proconvulsant agent. The mechanisms of potentiation of seizures induced by metrazol after the administration of the sweetner in a single rapid intake will be discussed. PMID- 2974655 TI - Adverse influence of recipient lymphoid resistance to in vitro immunosuppression on the outcome of kidney transplants. AB - The study investigated whether preoperative in vitro sensitivity of lymphocytes from potential renal transplant recipients could identify patients at increased risk of acute rejection following transplantation and immunosuppression with cyclosporine, azathioprine, and prednisolone. Mixed lymphocyte culture responses were measured preoperatively in the presence of methylprednisolone, CsA, and antithymocyte globulin, and without immunosuppressive agents in 50 transfused recipients of primary cadaver renal transplants. Patients were classified as sensitive if all three immunosuppressive agents produced more than 50% inhibition of their MLC responses, and as resistant if one or more agents failed to produce 50% inhibition. All patients received postoperatively a standardized triple immunosuppressive regimen. Acute rejection was confirmed histologically and treated with Pred with or without ATG or monoclonal antibody OKT3. A total of 29 patients (58%) were sensitive and 21 (42%) were resistant; 4 patients were resistant to 3 agents, 5 were resistant to MP and ATG, 6 were resistant to MP and CsA, and 6 were resistant to MP alone. Sensitive and resistant groups did not differ in age, sex, transfusion history, HLA A, B and DR mismatches or duration of follow-up. The resistant group had a higher rate of graft loss from acute rejection (chi 2 = 6.0, d.f. = 1, P less than 0.02), more episodes of acute rejection (chi 2 = 8.7, d.f. = 3, P less than 0.05), and a higher proportion of patients in whom reflux nephropathy was the cause of renal failure (chi 2 = 18.3, d.f. = 1, P less than 0.001). The resistant group also had a higher proportion of highly sensitized patients and higher serum creatinine concentrations than the sensitive group, although the differences did not reach statistical significance. The study indicates that patients at high risk of acute rejection of renal allografts can be identified by a pretransplant in vitro assay, a finding that could influence recipient selection and immunosuppression. PMID- 2974656 TI - Allogeneic lymphocyte stimulation by human spleen. Identification of cells that are very effective for presenting class II HLA antigens. AB - Human spleen cells were fractionated by percoll density gradient centrifugation and by sorting in the FACS with mixtures of fluorescent antibodies against T cells, B cells, monocytes, and Sig-bearing cells. Cells responsible for powerful MLR stimulation were class II HLA antigen-positive and were concentrated in preparations depleted of all the markers listed above. These cells represented 1 2% of the initial spleen cells. They were remarkably more active than other HLA class II antigen-positive cells. The procedure described allows rapid enrichment for the responsible cells. It should be useful for further characterization of these cells and for performing studies on their function. PMID- 2974657 TI - Inhibition of factor VIII synthesis by kidney endothelial cells by graft infiltrating T helper lymphocytes. PMID- 2974658 TI - [Clinical aspects of severe psoriasis]. PMID- 2974659 TI - [Electrogenic activity of Ca2+-ATPase fragments of the sarcoplasmatic reticulum during the early stages of radiation injury]. AB - It is established that local X-ray irradiation of the rabbit hind limb produces a decrease in Ca2+, Mg2+-ATP-dependent formation of electric potentials difference on the membrane of the sarcoplasmic reticulum (SR). These results agree with the observed decrease in the Ca2+-ATPase activity of SR membranes and increase in their electric conduction after irradiation. PMID- 2974660 TI - [Occupational damage to the back--accident or occupational disease]. PMID- 2974661 TI - [Cellular sensitization to Varidase]. PMID- 2974662 TI - [Exanthema of the face]. PMID- 2974663 TI - [Percutaneous transluminal angioplasty (PTA) of pelvic and lower limb arteries]. PMID- 2974664 TI - Control of naturally acquired bovine parasitic bronchitis and gastroenteritis with an oxfendazole pulse release device. AB - Four groups, each of six male Friesian calves, were set-stocked on separate 0.66 ha paddocks from May 7 until October 23 1986. Each of the animals in groups 1 and 4 was dosed with an oxfendazole pulse release bolus at turn out whereas the animals in groups 2 and 3 were left untreated. Parasite-free naive tracer calves were introduced into each paddock for a limited period 12 days after turn out and again at the end of the trial. No adverse reactions or clinical signs were observed in either of the groups of calves which received boluses. The development of clinical parasitic gastroenteritis in both the untreated groups necessitated the humane slaughter of two animals and emergency anthelmintic treatment of the remainder. The lower plasma pepsinogen concentrations, and lower faecal egg and larval counts and worm burdens post mortem, together with the absence of clinical signs of parasitic gastroenteritis and bronchitis in the treated calves, confirmed the high efficacy of the bolus treatment. PMID- 2974666 TI - [The work of rear-service hospitals deployed in the territory of the Turkmen SSR during World War II]. PMID- 2974665 TI - Coronary arteritis and aortoarteritis in the elderly males. A report of two autopsy cases with review of the literature. AB - Two elderly males died suddenly of acute myocardial infarction. Autopsy disclosed extensive and severe coronary arteritis, associated with aortitis. In most of all of the epicardial coronary arteries, fibrous thickening of the adventitia was remarkable and muscle layers of the media were nearly destroyed by infiltration of lymphocytes and plasma cells. The aorta showed adventitial thickening with destruction of the elastic layers of the outer media. In the literature, similar extensive coronary arteritis has been reported in five cases of Takayasu's arteritis. All cases, including ours, were elderly or middle-aged males who died suddenly of myocardial infarction without the antemortem detection of the underlying arteritis. These aspects differ from the typical Takayasu's arteritis and we suggest that these cases are considered as a separate entity. PMID- 2974667 TI - [Vitamin status of workers in the chromium industry]. AB - The content of A, E, C, B2 and PP vitamins and their metabolites in the blood and urine as well as microsigns of vitamin deficiency were studied in workers engaged in chrome industry. Microsigns of vitamin deficiency were observed along with low levels of vitamins. This appears to be associated with increased physiological requirements in vitamins due to specific work of chrome industry personnel. PMID- 2974668 TI - [Effect of low mercury concentrations on the natural resistance of the body]. PMID- 2974669 TI - [Androgens--their secretion in physiological and pathological conditions]. PMID- 2974670 TI - [Hypertrophic cardiomyopathy--diagnostic significance of the disproportion of myocardial hypertrophy examined by two-dimensional echocardiography]. AB - 34 patients with myocardial hypertrophy, mean age 44 years, were examined by two dimensional echocardiography and the degree of disproportion of the hypertrophy of the different left ventricular segments was analyzed. Besides the classic index--ratio between the septum and the posterior ventricular wall--a new index for the asymmetry of the hypertrophic process is proposed: the ratio between the thickest, segment and the thinnest segment. The new index is more sensitive in determining the asymmetrical hypertrophy. PMID- 2974671 TI - [Therapy of peptic ulcer. ATPase inhibitors]. PMID- 2974672 TI - [Laparoscopy--relevant indications]. PMID- 2974673 TI - [Experimental phages for differentiating nontypable methicillin-resistant Staphylococcus aureus]. AB - The possibility of using phages, isolated from the lysogenic cultures of methicillin-resistant staphylococci and modified in methicillin-resistant cultures of phage 85, for the differentiation of nontypable staphylococcal strains has been studied. The variants of phage 85 cannot be used for the determination of differences between the strains of methicillin-resistant cultures; they are not suited for typing. For this purpose the collection of phages isolated from lysogenic methicillin-resistant cultures should be used. PMID- 2974674 TI - [The functioning of a drained natural focus of leptospirosis in a nonchernozem area]. AB - The drainage of a natural focus of leptospirosis of the flood plain-swamp type, carried out over the period of 15 years, has led to changes in the species structure of small mammals and to an increase in the number of Leptospira carrying species. Nevertheless, like before drainage, the prevalent species and the main carrier of leptospires is still the root vole (Microtus oeconomus Pallas). The intensity of the epizootic process among small mammals at the final stage of drainage has proved to be similar to that at the initial stage. At the same time the cultivated fields created on the drained territory of the natural focus abound, by the time of harvesting, with small mammals among whom the intensive epizootic process takes place. PMID- 2974676 TI - Site specific mapping of chromosome 21 gene probes and the study of Down syndrome. PMID- 2974677 TI - Electrocardiographic standard for the diagnosis of right ventricular hypertrophy in complete right bundle branch block. PMID- 2974675 TI - The effect of ipratropium bromide on nasal hypersecretion induced by methacholine in patients with vasomotor rhinitis. A double-blind, cross-over, placebo controlled and randomized dose-response study. AB - The effect of various doses of ipratropium bromide aerosol on nasal hypersecretion induced by five concentrations methacholine was studied in 24 patients with vasomotor rhinitis and excessive watery nasal secretion. The volume of nasal secretion was greater with each of the five increasing doses of methacholine from 7.5 to 120 mg/ml. The median volume of nasal secretion was alike in all patients after administration of methacholine only and after treatment by placebo followed by methacholine. When the patients were treated with ipratropium bromide prior to administering methacholine the volume of secretion was reduced significantly. With doses of 40 micrograms and 100 micrograms of ipratropium to each nostril a similar reduction in the volume of secretion occurred but a still greater reduction by the application of 200 micrograms of ipratropium when compared with treatment by the placebo. While the volume of secretion increased with each increasing concentration of methacholine, a similar pattern of reduced secretion for each concentration of methacholine was seen with each greater concentration of ipratropium. In patients with vasomotor rhinitis, treatment with ipratropium bromide was found to reduce significantly the hypersecretion induced by methacholine when compared with treatment by the placebo. This reduction was greater with greater doses of ipratropium. PMID- 2974678 TI - Combined esophageal and pyloric atresia without tracheoesophageal fistula in Down's syndrome. PMID- 2974679 TI - Foetal adrenal steroids--initiation of human parturition. AB - The role of foetal adrenal DHAS in initiation of human parturition was studied by direct estimation of umbilical arterial blood samples and correlated with the oxytocin consumption during induced labour, Author found a closed relation between the above parameters and suggested that foetal adrenal activity might be involved in the initiation of human parturition. PMID- 2974680 TI - Long term experience in the treatment of advanced prostatic cancer with decapeptyl, compared to orchiectomy. PMID- 2974681 TI - The effects of syphilis on endocrine function of the fetoplacental unit. AB - Several pregnancy complications that are thought to cause chronic intrauterine stress have been found to lead to inappropriate fetal development and reductions in estrogen production. In the current study we sought to evaluate the fetoplacental unit in pregnancies complicated by maternal syphilis (n = 37), with and without fetal infection. Maternal 17 beta-estradiol and estriol levels were reduced during the third trimester in women with syphilis when compared with those in women with uncomplicated pregnancies. Serum progesterone levels were within normal limits or else were increased in women with syphilis. When compared with data in age- and weight-matched control infants of women having no pregnancy complications, umbilical cord serum levels of dehydroepiandrosterone sulfate, the major fetal adrenal precursor of placental estrogens, were subnormal (897 +/- 597 ng/ml, mean +/- SD) in 12 newborn infants with congenital syphilis (33.9 +/- 4.2 weeks' gestation, birth weight 2020 +/- 719 gm); such infants also had excessive serum levels of cholesterol (103 +/- 37 mg/dl). Dehydroepiandrosterone sulfate (1883 +/- 907 ng/ml) and cholesterol (58.1 +/- 13.9 mg/dl) levels were within normal limits in 19 uninfected infants of women with syphilis (38.6 +/- 2.4 weeks' gestation, birth weight 2861 +/- 660 gm). Cortisol levels were increased and estriol levels were decreased in both groups of neonates of women with syphilis compared with those in control neonates. These findings are suggestive that estrogen production often is reduced in pregnancies complicated by syphilis; the reduction in estriol appears to be largely due to reduced fetal adrenal dehydroepiandrosterone sulfate production. The reduction in 17 beta-estradiol levels may be due to alterations in maternal precursor synthesis. Although placental progesterone formation appears to be normal in women with syphilis, a deficiency in placental aromatase activity also is possible. PMID- 2974682 TI - Autonomic modulation of fetal atrial natriuretic factor and urine flow responses to vascular volume expansion. AB - The present study was designed to test the hypothesis that the fetal autonomic nervous system modulates the fetal plasma atrial natriuretic factor and urine flow responses to vascular volume expansion. To do this, isotonic saline solution (2% of estimated body weight) was infused intravascularly over a 10-minute period into autonomically intact or autonomically blocked fetal sheep. The autonomically intact fetuses responded to the infusion with a significant rise in arterial and venous pressures (p less than 0.0001), and atrial natriuretic factor increased by 36 +/- 8 (SE) pg/ml (p less than 0.05). In the autonomically blocked fetuses, the arterial pressure rise was greater (7.5 +/- 1.3 vs 2.6 +/- 0.7 mm Hg), whereas the venous pressure rise was not different from that seen in the intact fetuses. Atrial natriuretic factor increased by 44 +/- 20 pg/ml (p less than 0.05) in the blocked fetuses. Urine flow increased by similar amounts in the two groups (maximal increase above control of 0.33 +/- 0.11 in the intact and 0.27 +/- 0.10 ml/min in the blocked fetuses). Thus autonomic blockade did not significantly alter either the atrial natriuretic factor, urine flow, or venous pressure responses to volume loading in the fetal sheep. Therefore it appears that the autonomic nervous system does not play a major role in modulating atrial natriuretic factor, urine flow, or venous pressure responses to acute vascular volume expansion in the late gestational fetal sheep. PMID- 2974683 TI - Ovine fetal lung fluid response to intravenous saline solution infusion: fetal atrial natriuretic factor effect. AB - The fetal lung, a significant source of in utero fluid production, has been postulated to serve a regulatory role in maintenance of fetal body fluid homeostasis. Whereas the fetus responds to intravascular saline solution infusions with increased urine output, the fetal lung fluid response to this stimulus is unclear. Tracheal fluid output was measured in four chronically catheterized ovine fetuses (mean gestation, 129 +/- 1 days) subjected to successive 40-minute intravenous 0.9% saline solution infusions at rates of 0.5 and 1 ml/min/per kilogram of body weight. Tracheal fluid output decreased significantly (1.7 +/- 0.1 to 1.1 +/- 0.1 ml/10 min, p less than 0.01) during the infusion and returned to basal levels during the recovery period. Lung fluid osmolality and electrolyte concentration did not change. Fetal plasma atrial natriuretic factor increased significantly in response to the saline solution infusion (364 +/- 90 to 790 +/- 286 pg/ml, p less than 0.05) and returned to basal levels during the recovery period. There was a significant inverse correlation between plasma atrial natriuretic factor levels and tracheal fluid output. These results suggest that increased fetal plasma atrial natriuretic factor decreases lung fluid production. Lung fluid does not appear to compensate for fetal body water excess. Rather, lung fluid production appears to promote intrauterine pulmonary growth and to facilitate the transition to the extrauterine environment. PMID- 2974685 TI - Dose-response curves of the uterine and placental vascular beds to prostaglandin I2. AB - Local infusion of prostaglandin I2 (PGI2) has been reported to dilate the uteroplacental vasculature in a dose-dependent manner. In this experiment we attempted to distinguish the placental and nonplacental (uterine) components of this response over four concentrations of PGI2. Eleven near-term sheep were chronically instrumented for determination of regional blood flows by the use of radioactive microspheres. PGI2 was administered in a retrograde manner via a branch of the middle uterine artery at 1, 3, 10, and 20 micrograms/min. Flows were measured before (control) and after 5-minute infusions at each of the four concentrations (test). The uterine vasculature vasodilated in response to local PGI2 infusion. The 10 micrograms/min dose, for example, produced a mean (+/- SEM) flow of 0.70 +/- 0.07 ml/min/gm; the control value was 0.41 +/- 0.03 ml/min/gm (p less than 0.001). At 20 micrograms/min the test and control flows were 0.75 +/- 0.16 and 0.36 +/- 0.06 ml/min/gm (p less than 0.05), respectively. Uterine vascular resistance fell in a dose-dependent manner as well. There was no evidence of placental vasodilation at any of the doses tested. Renal vasodilation and decreased systemic arterial pressure at higher PGI2 doses suggest a recirculation effect. We conclude that PGI2 does not dilate the placental vasculature over the dose range of 1 to 20 micrograms/min and that the reported vasodilation of the uteroplacental vasculature is a result of decreased resistance in the uterine vasculature alone. PMID- 2974684 TI - Atrial natriuretic factor, digoxin-like immunoreactive substance, norepinephrine, epinephrine, and plasma renin activity in human fetuses and their alteration by fetal disease. AB - We measured five hormones presumably involved in fetal homeostasis in specimens obtained by cordocentesis for clinical indications from 106 fetuses. Norms for atrial natriuretic factor, digoxin-like immunoreactive substance, plasma renin activity, norepinephrine, and epinephrine were derived from fetuses ultimately shown to be free of detectable abnormality. Atrial natriuretic factor, digoxin like immunoreactive substance, and plasma renin activity were unrelated to umbilical vessel source or gestational age. Digoxin-like immunoreactive substance was directly related to PCO2 (r = 0.63, p = 0.02). Digoxin-like immunoreactive substance level was elevated in all fetal disease states studied except isoimmunization. The level of atrial natriuretic factor was elevated in fetuses with immune hydrops (NS). Norepinephrine and epinephrine levels were higher in the umbilical artery than in the vein (p = 0.05 and 0.006, respectively). There was a significant correlation between norepinephrine and gestational age in normal fetuses (r = 0.7637, p less than 0.025) and between both catecholamines and many of the respiratory blood gas measurements, with pH and PCO2 being the major determinants. Most disease states were associated with an elevated norepinephrine concentration. There was a negative correlation between plasma renin activity and base deficit (p less than 0.0001). Plasma renin activity was elevated in fetuses with idiopathic growth retardation and nonimmune hydrops (p less than 0.05 for each). In summary, fetal homeostasis as reflected by these five hormones was altered by a variety of disorders. With these baseline values the effects of direct or indirect fetal therapy can begin to be studied. PMID- 2974686 TI - Effects of prolactin and bromocriptine on physiological status of testis of the musk shrew (Suncus murinus L.). AB - This investigation was undertaken to study the effects of prolactin and bromocriptine on the testis of the musk shrew. None of these treatments had any effect on the weight of testis or on the accessory sex organs. Treatment of prolactin or bromocriptine failed to induce any change in esterified and free cholesterol content of the testis. No significant alterations were also recorded in the levels of macromolecules and in the levels of enzymes of the testis and the prostate gland. On the other hand, bromocriptine treatment resulted in an increase in the activity of Mg2+-ATPase and phospholipid:DNA ratio of the testis with a concomitant decrease in its DNA content. The absence of any change in the content of fructose in the ampullary gland, in the activity of beta-glucuronidase of the kidney, in cholesterol content of the testis, in diameters of seminiferous tubule and Leydig cell nucleus, in activities of acid phosphatase and beta glucuronidase of the testis and in the weight of accessory sex organs of prolactin and bromocriptine treated musk shrews suggests that prolactin does not play a significant role in regulating the testicular function of the musk shrew. PMID- 2974687 TI - Glia cells as immunoregulatory elements. Up- and down-regulatory activities of astrocyte clones. PMID- 2974688 TI - Suppressor cell regulation of chronic relapsing experimental allergic encephalomyelitis. PMID- 2974689 TI - Inhibition of passive allergic encephalomyelitis by sulfated polysaccharides. PMID- 2974690 TI - Changes in immune function in relapsing multiple sclerosis correlate with disease activity as assessed by magnetic resonance imaging. PMID- 2974691 TI - Interaction of endogenous opioids and developing T lymphocytes. PMID- 2974692 TI - Role of mast cells in peripheral nervous system demyelination. PMID- 2974693 TI - Immunology of multiple sclerosis. PMID- 2974694 TI - Pain quantum and wound healing: a comparison of interrupted inversion PDS and standard nylon sutures in abdominal skin closure. AB - Patients' postoperative pain experience during 5 days after hernia repairs or upper abdominal procedures was compared when skin closure was achieved by either full thickness interrupted sutures or subcuticular interrupted inversion sutures. Quantitative pain assessments were made using linear analogue charts and standardised analgesic requirements. Clinical advantage for the subcuticular technique was confirmed for the hernia group and possible advantage in the upper abdominal closure, suggested by lesser analgesic requirement. Statistical analysis has lent support to the hernia group findings but is less clear in the upper abdominal group. A comparison of wound healing, sepsis and cosmetic outcome showed no apparent advantage to either method. Economies and patient preference favoured the subcuticular method. PMID- 2974695 TI - [Treatment by redilatation and thrombolysis of occlusion occurring during percutaneous coronary angioplasty]. AB - In patients who develop acute coronary occlusion during or after percutaneous coronary angioplasty, surgery is not mandatory, and other treatments may be considered, namely redilatation and/or thrombolysis. Between June, 1984 and January, 1988 we performed 500 dilatations of coronary arteries, not counting the attempts made in the acute phase of myocardial infarction. Acute coronary occlusion without angiographic image of occlusive dissection occurred in 31 patients (6.2%) and was treated by attempted redilatation and intracoronary thrombolysis. In 10 patients (group A) either the occlusion could not be removed and emergency surgery was tried (5 cases with 2 infarctions and 1 death), or the occlusion was removed but myocardial infarction took place (5 cases). In 21 patients (group B), the occlusion was removed and the outcome was favourable without myocardial infarction. Altogether, myocardial infarction or death occurred in only 8 cases, or 26% of acute occlusions. The clinical and angiographic features of the two groups before and after angioplasty were compared; two of them differentiated group A from group B: (1) unstable angina, 7/10 in group A, 4/21 in group B (p less than 0.01), and (2) degree of stenosis, 93.1% in group A, 78% in group B (p less than 0.01). When coronary occlusion occurs during or after coronary angioplasty and is poorly tolerated with fall in blood pressure, surgery must be contemplated at once, even after recanalization of the vessel and subsidence of ischaemia. In all other cases, treatment with both redilatation and thrombolysis should restore the benefits of angioplasty without myocardial infarction. PMID- 2974696 TI - The human protein S locus: identification of the PS alpha gene as a site of liver protein S messenger RNA synthesis. AB - The protein S locus, situated on chromosome 3, consists of two protein S genes. Here, we report the cloning and complete nucleotide sequence of the 3' untranslated region of the two genes designated PS alpha and PS beta. Both regions span approximately 1,200 nucleotides. They show a high degree (-97%) of homology, with deviations caused by small deletions, insertions and point mutations. Comparison of PS alpha and PS beta with the reported protein S liver cDNAs, shows that the latter all originate from the PS alpha gene. The PS alpha gene therefore is marked as the major site of synthesis of liver protein S mRNA. Sequence comparison with the bovine protein S cDNA reveals that the PS beta gene has accumulated a few more mutations than the PS alpha gene since duplication of the ancestral protein S gene that seems to have occurred recently during primate evolution. PMID- 2974697 TI - Radiolabeling of the interferon-alpha receptor. AB - The action of alpha interferon (IFN-alpha) is initiated by its binding to a specific cell-surface glycoprotein, the IFN-alpha receptor, which is not well characterized. IFN-alpha A was reacted with an 125I-labeled, cleavable, heterobifunctional reagent. The derivatized IFN-alpha A was bound to human Daudi cells and photoactivated, forming a covalent IFN/receptor complex of apparent molecular weight 130,000-140,000 by SDS-polyacrylamide gel electrophoresis. Cleavage of the complex produced a new 125I-labeled 110 kDa band, representing the 125I-labeled IFN-alpha receptor free of IFN-alpha. This result provides a better estimate of the apparent molecular weight of the IFN-alpha receptor, and also provides a tool for tracking the migration of the free receptor in SDS polyacrylamide gel electrophoresis. PMID- 2974698 TI - Insulin-mediated translocation of glucose transporters from intracellular membranes to plasma membranes: sole mechanism of stimulation of glucose transport in L6 muscle cells. AB - Plasma membranes and light microsomes were isolated from fused L6 muscle cells. Pre-treatment of cells with insulin did not affect marker enzyme or protein distribution in isolated membranes. The number of glucose transporters in the isolated membranes was calculated from the D-glucose-protectable binding of [3H]cytochalasin B. Glucose transporter number was higher in plasma membranes and lower in intracellular membranes derived from insulin-treated cells than in the corresponding fractions from untreated cells. The net increase in glucose transporters in plasma membranes was identical to the net decrease in glucose transporters in light microsomes (2 pmol/1.23 x 10(8) cells). The fold increase in glucose transporter number/mg protein in plasma membranes (2-fold) was similar to the fold increase in glucose transport caused by insulin. This suggests that recruitment of glucose transporters from intracellular membranes to the plasma membrane is the major mechanism of stimulation of hexose transport in L6 muscle cells. This is the first report of isolation of the two insulin-sensitive membrane elements from a cell line, and the results indicate that, in contrast to rat adipocytes, there is not change in the intrinsic activity of the transporters in response to insulin. PMID- 2974699 TI - The amino acid sequence of the 24-kDa subunit, an iron-sulfur protein, of rat liver mitochondrial NADH dehydrogenase deduced from cDNA sequence. AB - Antiserum directed against bovine heart mitochondrial NADH dehydrogenase has been used to screen a rat liver cDNA expression library in lambda gt11. The insert cDNA of a positive clone was found to represent the 24-kDa subunit of NADH dehydrogenase by epitope selection using nitrocellulose filter containing the expressed proteins. The amino acid sequence deduced from the nucleotide sequence of the cloned cDNA indicated that the 24-kDa subunit is produced as a precursor with an amino-terminal extension, and that its mature form consists of 217 amino acid residues with a molecular weight of 23,933. PMID- 2974700 TI - Characterization of receptor tyrosine-specific protein kinases by the use of inhibitors. Staurosporine is a 100-times more potent inhibitor of insulin receptor than IGF-I receptor. AB - The receptors for insulin and insulin-like growth factor (IGF)-I carry intrinsic tyrosine-specific protein kinases (TPK) in their cytoplasmic domains that show 84% homology. Our previous studies using tyrosine-containing synthetic polymers (Arch. Biochem. Biophys. 260, 416, 1988) revealed subtle differences between the two receptor TPKs. In the present study, low molecular weight kinase inhibitors were used to compare the two receptor TPKs purified from human placenta. Staurosporine was the most potent inhibitor of both receptor TPKs among the three inhibitors tested. It was 100 times more inhibitory to insulin receptor TPK (ED50 = 61nM) than IGF-I receptor TPK (ED50 = 6.2 microM). Apigenin and kaempferol showed approximately the same inhibitory potency toward both TPKs with a range of 10 approximately 1000 microM. Staurosporine is thus an excellent tool to biochemically characterize the two receptor TPKs as well as to selectively inhibit insulin-activated TPK in intact cells. PMID- 2974701 TI - Molecular cloning of a cDNA for rat liver monoamine oxidase B. AB - The cDNA for rat monoamine oxidase B mRNA was isolated from liver cDNA library in lambda gt11 using specific antibody and oligonucleotide probes derived from FAD containing peptide of the enzyme. The primary structure of the protein, deduced from the nucleotide sequence, consisted of 520 amino acid residues and its molecular weight was calculated to be 58.4 kD which is in good agreement with that of the in vitro-synthesized peptide. FAD-binding site is located in the carboxy-terminal region. There is no typical structural feature common to the targeting signals for mitochondria, the periodic distribution of basic amino acids spaced by several uncharged residues, at its amino-terminal region. This region has an uninterrupted stretch of 14 hydrophobic residues. PMID- 2974702 TI - Clinical evaluation of a monophasic ethinylestradiol/desogestrel-containing oral contraceptive. AB - A multicenter trial was conducted in 267 centers in Italy to evaluate the efficacy, acceptability and safety of a monophasic oral contraceptive containing 30 micrograms ethinyl estradiol and 150 micrograms desogestrel (Marvelon) 13,290 women were followed up for a total of 74,967 cycles. No pregnancies due to method failure were reported and only three because of patient failure. Cycle control was good. There was a decrease in the occurrence of irregular cycles and in the duration and amount of menstrual bleeding; the incidence of breakthrough bleeding and spotting was low. No severe side-effects occurred and the incidence of minor complaints was generally lower during treatment than in the pre-treatment cycle. Body weight and blood pressure were not significantly altered. PMID- 2974703 TI - Molecular and immunologic analyses of a functional internal image formed by an anti-receptor antibody. PMID- 2974705 TI - Ventricular aberrant bands and hypertrophic trabeculations. A clinical pathological correlation. AB - We reviewed the clinical, hemodynamic, and pathology data of 22 patients with dilated congestive cardiomyopathy and 13 patients with ischemic heart disease who underwent heart transplantation, primarily to improve the pathological definition of left ventricular (LV) and right ventricular (RV) aberrant bands and hypertrophic trabeculations. Overall prevalence of aberrant bands was 37% in th LV and 28% in the RV. Similar values for hypertrophic trabeculations were 43% and 28%, respectively. Compared with ischemic heart disease, our patients with dilated congestive cardiomyopathy had similar ventricular size and wall thickness, but had a higher prevalence of LV aberrant bands (p = .005) and LV hypertrophic trabeculations (p = .01). Aberrant bands were associated, both in the LV and RV, with dilated cavities (p less than .05), whereas hypertrophic trabeculations were associated with more ventricular hypertrophy and smaller LV size. Following morphological and histological analysis of the aberrant bands, we propose their division into two categories: genuine or primary bands (probably congenital in origin) and secondary bands, which most probably represent trabecular structures that develop a free cavitary course following pathological changes in the ventricular wall structure and cavitary geometry. Compared with the muscular RV bands situated mostly in the distal portion of the ventricle, LV bands were usually fibrotic and had at least one point of insertion in the inflow or outflow tract. The pattern of trabecular hypertrophy was also different in the two ventricles. Ventricular arrhythmias and thrombi occurred equally in patients with and without bands or trabeculations. PMID- 2974704 TI - Serum IgG brain reactive antibodies in Alzheimer disease and Down syndrome. AB - The cause(s) of Alzheimer disease (AD) remain unknown. Since neurofibrillary tangles and neuritic plaques, a hallmark of AD, are also present in older patients with Down syndrome (DS), a close relationship between the two diseases has been suggested. IgG type of brain reactive antibodies (BRAs) are present in a majority of patients with AD but such studies have not been carried out in adult patients with DS. The present study was carried out to test the hypothesis that if the brain changes in older patients with DS and patients with AD are identical, the occurrence and specificity of BRAs should be similar in both diseases. The frequency of BRAs was studied in sera obtained from 21 patients with AD, 26 older patients with DS, and 20 normal control subjects using electroblots prepared from rat spinal cord proteins. Circulating BRAs were found in 57% of patients with AD, 81% of patients with DS, and 30% of normal controls. However, the pattern of BRA reactivity was different in AD and DS. In AD, antibodies reacted with the 200-kDa subunit of neurofilament triplet singly or in combination with the 150-kDa, 70-kDa or with the associated 62-kDa protein. In DS, BRAs always reacted with the 70-kDa protein, usually in combination with the 200-kDa subunit and, less frequently, in association with the other subunits. No specific pattern of antibody activity was evident in the control group. PMID- 2974707 TI - Mechanisms of vessel injury during percutaneous transluminal angioplasty of saphenous vein bypass grafts and coronary arteries. AB - Three patients were studied in whom percutaneous transluminal angioplasty (PTA) was attempted in stenotic saphenous vein bypass grafts. In two cases, large splits were present in the thick fibrotic intima of the vein grafts, accompanied by rupture of atherosclerotic plaques or hemorrhage into atherosclerotic plaques. Both cases required emergency coronary artery bypass surgery. In the third case, a vein graft failed to dilate after multiple attempts. Only small splits were present in that fibrotic intima. Two other patients were studied in whom coronary artery PTA was attempted. In both cases, laceration of a nonatherosclerotic segment of the artery occurred. Combined with our previous report, these observations indicate that PTA results in lacerations of atherosclerotic plaques, fibrotic intima, and otherwise normal vessel walls and that laceration is the mechanism of both successful dilation and serious complications. PMID- 2974706 TI - The enigma of restenosis after percutaneous transluminal coronary angioplasty. PMID- 2974709 TI - Analysis of absolute T helper cell number and cellular immune defects in HIV antibody positive and negative homosexual men. AB - To investigate if serial measurement of T helper (CD4) lymphocyte number in peripheral blood is of prognostic value, we determined lymphocyte function in asymptomatic and symptomatic HIV antibody positive and negative homosexual males and related the results to absolute number of CD4 lymphocytes in peripheral blood. Lymphocyte function was determined by measuring streptolysin O (SLO) induced proliferative responses of peripheral blood lymphocytes (PBL) and of PBL depleted of CD8 lymphocytes. Phytohemagglutinin (PHA) induced interleukin-2 (IL 2) production was also measured. In all functional tests values were significantly lower in HIV antibody-positive subjects than in HIV antibody negative subjects. Results lower than the 95% confidence limit in HIV antibody negative individuals were therefore defined as "decreased." Decreased functional responses were most frequent (83-100%) in individuals with a number of CD4 lymphocytes of less than 400/microliters, and were least frequent (3-21%) in subjects with a CD4 lymphocyte count of greater than 600/microliters. Frequency of decreased functional responses was intermediate in the population with 600 400/microliters CD4 lymphocytes. The magnitude of functional responses differed significantly between groups with less than 400, 400-600, and greater than 600 CD4 lymphocytes per microliter, indicating that T helper cell number decreases with loss of immune function. PMID- 2974708 TI - Inhibition of late influenza virus genome expression by diamidinophenylindole. AB - The growth cycle of influenza virus strain FPV, Ulster 73, was altered by treatment of LLC-MK2 cells with diamidinophenylindole. Viral protein synthesis was restricted to the early pattern of virus multiplication, and post-treatment experiments showed the ability of the drug to block virus replication until the 4th hour p.i. Drug addition (followed by removal) revealed the inhibition of synthesis of late viral products, and especially of membrane protein. Kinetic studies on the production of viral RNA indicated a decrease in the synthesis of late virus-induced RNA species, suggesting that the target of DAPI is probably the late transcription of the virus genome. The nonpermissive condition mediated by the drug could represent a suitable model to study cellular intervention during viral growth. PMID- 2974710 TI - Can social workers damage your dental health? PMID- 2974711 TI - An elective in San Francisco. HIV infection: comparing attitudes, knowledge and cross-infection control amongst dentists in San Francisco and Great Britain. PMID- 2974712 TI - Do we need more muscle relaxants? PMID- 2974713 TI - Sufentanil and nitrous oxide anaesthesia for cardiac surgery. AB - We have investigated the use of sufentanil 3.75-15 micrograms kg-1 by supplementing anaesthesia with nitrous oxide and midazolam. Thirty patients with ejection fractions exceeding 30% were studied while undergoing scheduled coronary artery vein graft surgery. Even in the lowest dose group (3.75 micrograms kg-1), haemodynamic responses to surgical and anaesthetic stimuli were sufficiently obtunded that no patient exhibited an increase in heart rate or systolic arterial pressure greater than 20% of the control value. Marked hypotension occurred in some patients during unstimulated periods. Such periods of hypotension were associated with equally marked decreases in systemic vascular resistance. The mean recovery times to spontaneous ventilation after the end of surgery ranged from 6 to 12 h. This is longer than would be expected from other studies using a similar dose of sufentanil. This may be related to the use of benzodiazepines during anaesthesia and to their use after surgery in those patients who became restless. PMID- 2974714 TI - Adverse reactions to atracurium and alcuronium. A prospective surveillance study. AB - A multicentre prospective surveillance study was undertaken to compare the incidence and severity of adverse reactions attributed to atracurium and alcuronium. Clinical manifestations were used by the anaesthetist to diagnose an adverse reaction (a cutaneous reaction, a greater than 20% change in arterial pressure or heart rate, and bronchospasm). Of the 1956 patients receiving atracurium, 10.1% had adverse reactions compared with 17.9% of the 1425 patients receiving alcuronium (P less than 0.001). There were no longterm sequelae. The atracurium group had a markedly lower incidence of hypotension (3.4% v. 13.7%; P less than 0.0001), but a higher incidence of cutaneous reactions (4.6% v. 2.3%; P less than 0.005) which were not associated with other adverse reactions. There was a low incidence of bronchospasm in both groups (0.2% v. 0.1%). PMID- 2974716 TI - Cholecystocutaneous fistula. PMID- 2974715 TI - Interactions between cilazapril and propranolol in man; plasma drug concentrations, hormone and enzyme responses, haemodynamics, agonist dose-effect curves and baroreceptor reflex. AB - 1. The pharmacokinetics, hormonal and haemodynamic responses at rest and during challenges with angiotensin I (blood pressure), isoprenaline (heart rate), and noradrenaline (blood pressure) were investigated in six healthy male volunteers following a 1 week treatment with placebo, propranolol (120 mg day-1), cilazapril (2, 5 mg day-1), and a combination of both in a double-blind cross-over design. 2. Both drugs reduced systolic and diastolic blood pressure by about 7 mm Hg as compared with placebo. After coadministration, this drop in blood pressure was doubled and lasted longer than after the administration of the individual components. 3. Following cilazapril, a pronounced increase in plasma renin activity (PRA) was found (factor approximately 10 at drug peak concentrations). Coadministration of both drugs resulted only in a moderate increase in the PRA (factor approximately 3). Significant changes in plasma catecholamines were not observed. 4. Propranolol shifted the isoprenaline dose-effect curve to the right, and cilazapril that of angiotensin I, irrespective of the presence of the other drug. Cilazapril tended to shift the noradrenaline dose-effect curve somewhat to the right. 5. The gain of the baroreceptor reflex (angiotensin-stimulation) was not influenced by cilazapril but was lowered by propranolol, irrespective of the presence of the ACE inhibitor. 6. Except for a statistically not significant decrease in the peak concentrations of each drug during the combined therapy, a pharmacokinetic interaction between the two drugs was not found. PMID- 2974718 TI - Skeletal toxicity with isotretinoin therapy: a clinico-radiological evaluation. AB - Skeletal toxicity is known to occur with high doses of isotretinoin (greater than 2 mg/kg/day). We have attempted to evaluate the clinical significance and document the extent of musculoskeletal toxicity associated with a relatively low dose of isotretinoin (0.5 mg/kg/day) used in the treatment of severe acne. Radiographs of 120 patients were examined. Twelve per cent showed minor changes (four patients had spinal hyperostoses and 10 had calcaneal hyperostoses). None of the musculoskeletal changes we observed was clinically significant. Comparison with matched control X-rays showed 8% of the controls to have similar non significant changes. Follow-up of 11 of the patients with abnormal X-rays showed minor deterioration in one patient, no change in four and improvement in six. Thus, doses of 0.5 mg/kg/day isotretinoin in such patients did not produce any significant long-term musculoskeletal changes. With increasing use of this beneficial drug in acne, radiologists and dermatologists should be aware of its skeletal toxicity. PMID- 2974717 TI - High dose ketoconazole: endocrine and therapeutic effects in postmenopausal breast cancer. AB - Ketoconazole, an antifungal agent, inhibits in vitro C17-C20 lyase, an enzyme involved in androgen biosynthesis. Since adrenal and ovarian androgens are the main precursors of oestrogens in postmenopausal women, the endocrine and therapeutic effects of high dose ketoconazole (400 mg three times a day) were evaluated in 14 postmenopausal women with advanced breast cancer. Testosterone levels were suppressed significantly (37%, P less than 0.025), as was dehydroepiandrosterone sulphate, and androstenedione levels showed a similar but non-significant fall. Seventeen hydroxyprogesterone levels rose significantly, as would be expected if C17-C20 lyase was inhibited. There was no suppression of cortisol or oestrone levels. There was a small suppression of oestradiol concentrations, reflecting a decrease in its precursor, testosterone. Sex hormone binding globulin levels rose, which may be due to a decrease in testosterone. All the changes are compatible with C17-C20 lyase as a major site of action in vivo. No responses occurred in 12 patients treated with ketoconazole alone, but in 2 patients who were progressing on aminoglutethimide, testosterone levels were suppressed and in one patient a partial response occurred. Ketoconazole was poorly tolerated due to gastrointestinal toxicity. This study shows that C17-C20 lyase is a potential target for hormone therapy, and that sequential blockade of enzymes involved in oestrogen biosynthesis should be further evaluated. PMID- 2974719 TI - A comparative study of benzoyl peroxide and clindamycin phosphate for treating acne vulgaris. AB - A water based 5% benzoyl peroxide gel (Benzac W5) was compared with topical 1% clindamycin phosphate solution (Cleocin T) in the treatment of acne vulgaris using a randomized, investigator blind study design. Lesion counts were significantly reduced in both treatment groups over the 12-week study period; however, the reduction of total lesions produced by benzoyl peroxide gel was significantly greater than that produced by clindamycin phosphate (P less than 0.05). Clindamycin phosphate had a milder effect on the skin surface in terms of peeling and drying than the benzoyl peroxide gel. PMID- 2974720 TI - Acne vulgaris and malignancy. AB - Seventy patients with lymphomas and leukaemias were interviewed. Details of past acne vulgaris were recorded and a clinical examination performed. A history of acne was less common in the patients than in matched controls. In addition, fewer patients had received treatment prescribed by a medical practitioner for their acne than had the controls. Examination revealed fewer patients with acne. These results suggest a reduced incidence of acne or its more severe forms in patients who have developed lymphomas and leukemias, and provide some support for the hypothesis that increased exposure or response to P. acnes, as appears to occur with acne, could confer protection against malignant disease. PMID- 2974722 TI - Mechanism-based inactivation of rabbit muscle phosphoglucomutase by nojirimycin 6 phosphate. AB - Nojirimycin 6-phosphate (N6P) was tested as a substrate and inhibitor for phosphoglucomutase (PGM). In the absence of glucose 1,6-bisphosphate (GBP), the incubation of PGM and N6P resulted in the complete inactivation of all enzyme activity. When equimolar amounts of N6P and GBP were incubated together with PGM, the GBP was quantitatively converted to glucose 6-phosphate (G6P) and phosphate. At higher ratios of GBP and N6P (greater than 100) the final concentration of G6P produced was found to be 19 times the initial N6P concentration. These results have been interpreted to suggest that the phosphorylated form of PGM catalyzes the phosphorylation of N6P at C-1. This intermediate rapidly eliminates phosphate to form an imine and the dephosphorylated enzyme. The dephosphorylated enzyme is rapidly rephosphorylated by GBP and forms G6P. The imine is nonenzymatically hydrated back to N6P. Occasionally (5%) the imine isomerizes to a compound that is not processed by PGM. PMID- 2974721 TI - ABC excinuclease incises both 5' and 3' to the CC-1065-DNA adduct and its incision activity is stimulated by DNA helicase II and DNA polymerase I. AB - CC-1065 is a large molecule that binds covalently to adenine residues of DNA in a sequence-specific manner and lies in the minor groove about four bases to the 5' side of the adducted residue. Using a reconstituted Escherichia coli nucleotide excision repair system, we have obtained data showing that the ABC excinuclease makes incisions both 5' and 3' to the CC-1065 adduct and that the incision activity is stimulated by the addition of helicase II and DNA polymerase I (and dNTPs). Our results with the CC-1065 adduct are consistent with the reported in vitro processing of other adducts (e.g., cisplatin, UV photoproducts) but do not agree with a recent study that reported anomalous processing of the CC-1065 adduct by ABC excinuclease and helicase II. Our results also imply that, in binding to damaged DNA, ABC excinuclease does not make important contacts in the minor groove four bases to the 5' side of the damaged residue. PMID- 2974723 TI - Kinetic analysis of covalent hybrid plasminogen activators: effect of CNBr degraded fibrinogen on kinetic parameters of Glu1-plasminogen activation. AB - The kinetic parameters of three activator species of Glu1-plasminogen (Glu1-Plg) were compared in their reaction at pH 7.4 and 37 degrees C, in the presence and absence of CNBr-digested fibrinogen (CNBr-Fg). The urokinase- (u-PA-) derived covalent hybrid activator PlnA-u-PAB had an apparent Michaelis constant (Kplg) of 7.44 microM, a catalytic rate constant (kplg) of 51.1 min-1, and a second-order rate constant (kplg/Kplg) of 6.87 microM-1 min-1. The tissue plasminogen activator (t-PA) derived covalent hybrid activator PlnA-t-PAB was characterized by a Kplg of 3.33 microM, a kplg of 1.03 min-1, and a kplg/Kplg of 0.309 microM-1 min-1. The kplg/Kplg values for the parent u-PA and t-PA activators were 6- and 16-fold higher than the respective hybrids, mainly due to an approximately 10 fold increase in the apparent Kplg for the hybrids. In the presence of CNBr-Fg, the increase of the kplg/Kplg values for u-PA and its hybrid was 1.1-fold, but for t-PA and its hybrid, the increases were 7- and 12-fold, respectively. In both the absence and presence of CNBr-Fg, activator t-PAB had an apparent Kplg of 19.1 and 27.6 microM and a kplg of 2.9 and 5.0 min-1, respectively. The increase in the kplg/Kplg value with CNBr-Fg was 1.2-fold. The streptokinase- (SK-) derived activators Glu1-plasmin.SK (Glu1-Pln.SK), Val442-Pln.SK, and Val561-Pln.SK had apparent Kplg values of 0.458, 0.268, and 0.121 microM and kplg values of 20.0, 126.0, and 63.3 min-1, respectively. In the presence of CNBr-Fg, the first two activators showed an approximately 1.4-fold increase and the last showed a 1.4 fold decrease in their kplg/Kplg values. The catalytic efficiency (kplg/Kplg) of the various activator species fell in the decreasing order SK greater than u-PA greater than t-PA, in either the presence or absence of CNBr-Fg. CNBr-Fg enhanced significantly the activities of only two activators, t-PA and PlnA-t-PAB. PMID- 2974724 TI - Abnormal response to calmodulin in vitro of dystrophic chicken muscle membrane Ca2+-ATPase activity. AB - A skeletal muscle membrane fraction enriched in sarcoplasmic reticulum (SR) contained Ca2+-ATPase activity which was stimulated in vitro in normal chickens (line 412) by 6 nM purified bovine calmodulin (33% increase over control, P less than 0.001). In contrast, striated muscle from chickens (line 413) affected with an inherited form of muscular dystrophy, but otherwise genetically similar to line 412, contained SR-enriched Ca2+-ATPase activity which was resistant to stimulation in vitro by calmodulin. Basal levels of Ca2+-ATPase activity (no added calmodulin) were comparable in muscles of unaffected and affected animals, and the Ca2+ optima of the enzymes in normal and dystrophic muscle were identical. Purified SR vesicles, obtained by calcium phosphate loading and sucrose density gradient centrifugation, showed the same resistance of dystrophic Ca2+-ATPase to exogenous calmodulin as the SR-enriched muscle membrane fraction. Dystrophic muscle had increased Ca2+ content compared to that of normal animals (P less than 0.04) and has been previously shown to contain increased levels of immuno- and bioactive calmodulin and of calmodulin mRNA. The calmodulin resistance of the Ca2+-ATPase in dystrophic muscle reflects a defect in regulation of cell Ca2+ metabolism associated with elevated cellular Ca2+ and calmodulin concentrations. PMID- 2974725 TI - Pre-steady-state studies of the adenosine triphosphatase activity of coupled submitochondrial particles. Regulation by ADP. AB - ATPase activities were measured in 10 mM MgCl2, 5 mM ATP, 1 mM ADP, and 1 microM FCCP with submitochondrial particles from bovine heart that had been stimulated by delta mu H+-forming substrates and with particles whose natural inhibitor protein was partially removed by heating. The activities were not linear with time. With both particles, the rate of ATP hydrolysis in the 7-fold greater than that in the steady state. Pre-steady-state and steady-state kinetic studies showed that the decrease of ATPase activity was due to the binding of ADP in a high-affinity site of the enzyme (K0.5 of 10 microM). Inhibition of ATP hydrolysis was accompanied by the binding of approximately 1 mol of ADP/mol of particulate F1; 10 microM ADP gave half-maximal binding. ADP could be replaced by IDP, but with an affinity 50-fold lower (K0.5 of 0.5 mM). Maximal inhibition by ADP and IDP was achieved in less than 5 s. Inhibition was enhanced by uncouplers. Even in the presence of pyruvate kinase and phosphoenolpyruvate, the rates of hydrolysis were about 2.5-fold higher in the first seconds of reaction than in the steady state. This decrease of ATPase activity also correlated with the binding of nearly 1 mol of ADP/mol of F1. This inhibitory ADP remained bound to the enzyme after several thousand turnovers. Apparently, it is possible to observe maximal rates of hydrolysis only in the first few catalytic cycles of the enzyme. PMID- 2974726 TI - Fluorescence measurement of the kinetics of DNA injection by bacteriophage lambda into liposomes. AB - Bacteriophage lambda attaches to Gram-negative bacteria using the outer membrane protein LamB as its receptor. Subsequently, DNA is injected by the bacteriophage into the host cell for replication and expression. The mechanism of DNA injection, however, is poorly understood. In order to begin to characterize DNA injection, a quantitative kinetic assay to detect injection into reconstituted LamB liposomes is described. The technique involves monitoring the increase in fluorescence of liposome-encapsulated ethidium bromide, which occurs as DNA enters the aqueous compartment of the vesicles. The data indicate that injection is several times faster than indicated by earlier studies and is complete within 1 min. Such assays which allow direct observation of this process are necessary first steps toward a mechanistic understanding. PMID- 2974727 TI - Inhibition of membrane erythrocyte (Ca2+ + Mg2+)-ATPase by hemin. AB - Red blood cell lysis is a common symptom following severe or prolonged oxidative stress. Oxidative processes occur commonly in sickle cells, probably mediated through denatured hemoglobin and the accumulation of ferric hemes in the membranes. Calmodulin-stimulated (Ca2+ + Mg2+)-ATPase from sickle red cell membranes is partially inactivated (Leclerc et al. (1987) Biochim. Biophys. Acta 897, 33-40). In this study (Ca2+ + Mg2+)-ATPase activity from normal adult erythrocyte membranes was measured in the presence of hemin. We report a time- and concentration-dependent inhibition of the activity of the enzyme by hemin due to a decrease in the maximum velocity. Only a mild inhibitory effect was observed in the presence of iron-free protoporphyrin IX, indicating the catalytic influence of the iron. Experiments carried out with hemin (ferric iron) liganded with imidazole or with reduced protoheme (ferrous iron) liganded with carbon monoxide, demonstrated that the inhibition requires that hemin be capable of binding additional ligands. The inhibition was not influenced by the absence of oxygen but was prevented by addition of bovine serum albumin. Addition of butylated hydroxytoluene, a protective agent of lipid peroxidation, failed to prevent the inhibition of calmodulin-stimulated (Ca2+ + Mg2+)-ATPase. As dithiothreitol partially restores the enzyme activity, we postulated that hemin interacts with the thiol groups of the enzyme. PMID- 2974728 TI - Affinity labeling of the active site of the Ca2+-ATPase of sarcoplasmic reticulum. AB - The inactivation of sarcoplasmic reticulum ATPase by fluorescein isothiocyanate (FITC) was shown to have a hyperbolic dependence on the concentration of FITC. The results were quantitatively accounted for by a model in which the reagent first binds reversibly (Kf = 70 microM) to the ATPase and then reacts irreversibly (kmax = 0.8 and 2 min-1 in the absence and presence of 1 mM Mg2+, respectively) to form inactive enzyme. Comparison with the rate constant for the reaction of the model compound alpha-acetyllysine with FITC showed that the FITC reactive lysyl side-chain of the ATPase is not unusually reactive, indicating that the specificity of the reaction is due to affinity labeling behavior of the reagent. This was supported by protection experiments using ATP, ADP, AdoPP[NH]P, ITP, and TNP-ATP, all of which displayed protection constants similar to their known binding constants to the active site of the ATPase. Both inorganic phosphate and orthovanadate were effective in preventing inactivation by FITC, and calcium only partially reversed the effect of these anions, implying the existence of a ternary complex such as Ca2.E.Pi. Since all ligands (ATP, ADP and Pi) which bind or react at the catalytic site protect it, only the unliganded form appears to bind and react with FITC. Addition of calcium to the MgATP complex of the ATPase caused an increase in the FITC inactivation rate, implying that during turnover there is a larger fraction of unliganded enzyme present, i.e., substrate binding is weaker (Ks is larger). Protection was also observed with fluorescein and two related dyes, eosin and erythrosin. Like FITC, the isothiocyanates of these dyes were effective inactivators. In separate experiments, these two dyes were shown to promote photoinactivation of the ATPase. ATP exerted a protective effect with a concentration dependence consistent with high-affinity active-site binding. PMID- 2974729 TI - Chitin synthetase activity is bound to chitosomes and to the plasma membrane in protoplasts of Saccharomyces cerevisiae. AB - The sub-cellular distribution of chitin synthetase was studied in homogenates of Saccharomyces cerevisiae protoplasts. Use of a mild disruption method minimized rupture of vacuoles and ensuing contamination of subcellular fractions by vacuolar proteinases. After fractionation of whole or partially purified homogenates through an isopycnic sucrose gradient chitin synthetase activity was found to be distributed between two distinct particulate fractions with different buoyant density and particle diameter. When whole homogenates were used, about 52% of the chitin synthetase loaded was localized in a microvesicular population identified as chitosomes (diameter 40-110 nm; buoyant density (d) = 1.146 g/cm3). Another vesicular population containing 26% of the activity was identified as plasma membrane vesicles because of its large mean diameter (260 nm), its high buoyant density (d = 1.203 g/cm3) and by the presence of the vanadate-sensitive ATPase activity. Moreover, after surface labeling of protoplasts with 3H concanavalin A, the label cosedimented with the presumed plasma membrane vesicles. There was a negligible cross-contamination of the chitosome fraction by yeast plasma membrane markers. In both the plasma membrane and the chitosome fractions, the chitin synthetase was stable and essentially zymogenic. Activation of the chitosome fraction produces microfibrils 100-250 nm in length. Our results support the idea that chitosomes do not originate by plasma membrane vesiculation but are defined sub-cellular organelles containing most of the chitin synthetase in protoplasts of Saccharomyces cerevisiae. PMID- 2974730 TI - DNA polymerase alpha-DNA primase from human lymphoblasts. AB - The DNA polymerase alpha-DNA primase complex from the human lymphoblast line HSC93 has been enriched to near homogeneity by using an immunoaffinity purification protocol which was developed earlier for the purification of the calf thymus enzyme (Nasheuer, H.-P. and Grosse, F. (1987) Biochemistry 26, 8458 8466). Immunoaffinity purified polymerase-primase from human cells consisted of four subunits displaying molecular weights of 195,000 and 180,000 for the DNA synthesizing alpha-subunit, of 68,000 for the beta-subunit, and of 55,000 and 48,000 for the primase-carrying gamma- and delta-subunit, respectively. The isoelectric pH values for the individual subunits were estimated from non equilibrium pH gradients to be between 5.9 and 5.7 for the alpha-subunit, at 5.5 for the beta-subunit, and at 7.5 and 8.0 for the gamma- and delta-subunit, respectively. The purified polymerase-primase converted single-stranded phi X174 DNA into the double-stranded form in a primase-initiated reaction. During this process, 3-10 RNA primers were formed. RNA primers were about 11 nucleotides long. Elongation of existing RNA primers by the human polymerase-primase was semi processive; following primer binding the DNA polymerase continuously incorporated 20 to 50 nucleotides, then it dissociated from the template DNA. PMID- 2974731 TI - Effects of photoperiod, beta-endorphin, and naloxone on in vitro secretion of testosterone in white-footed mouse (Peromyscus leucopus) testes. AB - The effects of the pro-opiomelanocortin-derived beta-endorphin (B-EP) and the opioid antagonist naloxone on in vitro secretion (accumulation of testosterone (T) in the medium) of T by testicular cells were assessed in adult white-footed mice (Peromyscus leucopus). Animals were housed under long days (16L:8D) to maintain testicular function or under short days (8L:16D) to induce gonadal regression. In vitro treatment with B-EP or naloxone did not affect basal secretion of T in dispersed cells from active or regressed testes. However, B-EP caused a dose-dependent reduction in secretion of T from cells stimulated maximally with human chorionic gonadotropin (hCG) or dibutyryl cyclic adenosine 3', 5'-monophosphate (dbcAMP). Conversely, naloxone enhanced maximal hCG- and dbcAMP-stimulated secretion of T in testicular incubates from both long- (1.5 fold) and short-day (3.5-fold)-exposed mice. The finding that the addition of naloxone to maximally stimulated cells increased further the secretion of T is evidence that B-EP may act to inhibit gonadotropin-stimulated secretion of T. Also, the stimulatory effect of naloxone on cells from regressed testes indicates that B-EP may be involved in suppressing production of T during the gonadally regressed state. Testicular B-EP-like immunostaining is present within the cytoplasm of interstitial cells and is not apparent in the seminiferous tubules. Together, these results support the idea that in P. leucopus endogenous opioid peptides in the testes may aid in the regulation of testicular function throughout the yearly breeding cycle. PMID- 2974732 TI - Immunocytochemical localization of luteinizing hormone-releasing hormone and proopiomelanocortin neurons within the preoptic area and hypothalamus of the bovine brain. AB - The anatomical locations of proopiomelanocortin (POMC) and luteinizing hormone releasing hormone (LHRH) neurons were examined in brain tissue from peripubertal female calves and from mature, luteal-phase cows. Biotin-avidin-peroxidase procedures were used for single- and double-labeled immunostaining. LHRH perikarya formed a loosely arranged continuum, extending posteriorly from the diagonal band of Broca, passing through the medial and lateral preoptic areas, and ending within the anterior hypothalamic area. LHRH fibers, apparently directed toward the median eminence, passed (1) posteroventrally in the periventricular area and through the arcuate nucleus, and (2) ventromedially lateral to the arcuate nucleus and medial to the supraoptic nucleus. POMC perikarya were located within and about the arcuate nucleus, some penetrating into the median eminence. Fibers from these POMC perikarya passed ventrally to terminate in the median eminence and to regions ventral to the mammillary nuclei. POMC fibers also projected dorsally and laterally from the arcuate nucleus to other hypothalamic regions as well as anteriorly along the third ventricular wall to the preoptic area, bed nucleus of stria terminalis, and stria terminalis. Less than 6% of the POMC or LHRH processes were in close anatomical apposition to LHRH perikarya and dendrites. Extensive intermingling of LHRH and POMC fibers occurred within zona externa of the median eminence. PMID- 2974733 TI - [The action of nicotinamide on the adenine nucleotide system as well as on mitochondrial oxidation and phosphorylation processes in the liver of db/db strain mice]. AB - Both levels of total adenine nucleotides, ATP, AMP, ATP/ADP ratio and phosphate potential of cell and cytosol and the intensity of mitochondrial oxidation (fatty acid beta-oxidation in particular) and phosphorylation are elevated in the liver of db/db mice as compared with control. Presumably these alterations corresponding to the total activation of metabolic processes in db/db mice are mediated by hyperinsulinemia. Nicotinamide treatment (2.5 mg/100 g body weight, 14 days, i.m.) elicits further increase of ATP and total adenine nucleotide levels, cytosolic phosphate potential and activation of mitochondrial oxidation and phosphorylation. The findings obtained can be used for explanation of nicotinamide inhibition of gluconeogenesis, diacylglycerol and phosphoacylglycerol biosynthesis in the liver of db/db mice. PMID- 2974734 TI - [The viroimmunotest for mass determinations of nanogram amounts of antigens]. AB - Simple highly sensitive screening variant of viral immunoassay for detection of nanogram quantity of antigen is proposed. The antigen linked with antibodies adsorbed on polystyrene 96-well plates was revealed by conjugate of Fab' fragments of antibodies with phage phi X174. The concentration of the antigen could be detected qualitatively and quantitatively (from 0.3 ng/ml to 1000 ng/ml). PMID- 2974735 TI - [Ultrastructural changes in transverse striated muscles under the influence of space flight factors]. AB - The influence of space flight (on the biosatellite "Kosmos-1667") on muscles (diaphragmatic, soleus, gastrocnemius) was studied by electron microscope. Muscles had destructive and atrophic changes. The rate of changes was maximal in m. soleus, minimal in the diaphragmatic m. However, some regeneration was found demonstrating the reversibility of changes. PMID- 2974736 TI - Phage-plasmid hybrid vectors. PMID- 2974737 TI - Molecular cloning vectors for Aspergillus and Neurospora. PMID- 2974738 TI - Eleventh Gaddum memorial lecture. Lipocortin and the mechanism of action of the glucocorticoids. PMID- 2974739 TI - Anti-vasoconstrictor effects of the K+ channel opener cromakalim on the rabbit aorta--comparison with the calcium antagonist isradipine. AB - 1. Contractile responses of rabbit aortic rings elicited by KCl-depolarization, angiotensin II (AII), 5-hydroxytryptamine (5-HT) and noradrenaline (NA) have been investigated in the presence of cromakalim (BRL 34915) and isradipine (PN 200 110). 2. Above 10(-6)M, cromakalim inhibited contractile responses to low (less than or equal to 32 mM) but not to higher KCl concentrations. The 5-HT and AII concentration-response curves were antagonized noncompetitively by cromakalim (10(-7)-10(-5)M) and the maximal responses were inhibited by 40 and 55%, respectively. 3. Isradipine caused less inhibition of AII and 5-HT contractile responses than cromakalim, and in the presence of isradipine (10(-7)M), cromakalim was still able to antagonize further the contractions to AII in this vessel. 4. NA-induced contractions were relatively insensitive to inhibition by cromakalim and isradipine, both drugs causing a small rightward shift of the NA concentration-response curve. This result suggests that NA utilizes different Ca2+ pools from those involved in AII- and 5-HT-induced contractions of this vessel. 5. The sustained (tonic) part of the NA response was inhibited in a concentration-dependent manner by cromakalim (10(-7)-10(-5)M), but not by isradipine. 6. In aortic rings partially depolarized with 3.5 x 10(-2)M KCl, the ability of cromakalim, but not of sodium nitroprusside, atriopeptin III or hydralazine, to inhibit AII- and tonic NA-induced contractions was abolished. 7. Antivasoconstrictor activity of cromakalim on the rabbit aorta appears to involve factors in addition to an indirect inhibition of Ca2+ entry through dihydropyridine-sensitive Ca2+ channels. 8. The ability of cromakalim to open K+ channels and thereby modify the membrane potential would appear to underlie these antivasoconstrictor effects. This mechanism of action of cromakalim clearly differs from that of other vasodilators such as sodium nitroprusside and hydralazine. PMID- 2974742 TI - Assessment of angioplasty balloon catheters: preliminary studies. AB - Gruntzig balloon catheters from three manufacturers have been tested in vitro to establish a method of assessing the diameters and bursting pressures of inflated balloons. The method of testing and of measuring the inflated balloons is described. It was found that there were considerable variations in the behaviour of different balloons, and the relevance of this to arterial rupture is discussed. There were significant associations between the manufacturer and premature rupture of the balloon (p less than 0.01), and between co-axial construction of the catheter and overdistension of the balloon by more than 20% (p less than 0.01). The balloon length and the stated maximum diameter were not associated with either premature failure or overdistension. PMID- 2974741 TI - Ventral tegmental area: site through which dopamine D2-receptor agonists evoke behavioural and electrocortical sleep in rats. AB - 1. In freely moving rats the effects on behaviour and electrocortical (ECoG) spectrum power of some dopamine agonists, i.e. apomorphine and (+)-3PPP, given directly into different areas of the rat brain were studied. In particular, dopamine agonists were microinfused in the ventral tegmental area (VTA) and substantia nigra (SN) or into the caudate nucleus, n. accumbens and prefrontal cortex. The ECoG spectrum power effects were continuously analysed by means of a computerized Berg-Fourier analyser as total spectrum power and power in preselected frequency bands. 2. Apomorphine and (+)-3PPP (0.01, 0.1 and 1.0 nmol) given bilaterally into the VTA produced behavioural and ECoG sleep in a dose dependent fashion. A statistically significant (P less than 0.01) increase in ECoG total spectrum power with a predominant increase in the lower frequency bands (0.25-3, 3-6 and 6-9 Hz) occurred. No behavioural and ECoG changes were evoked by the same doses of apomorphine bilaterally microinfused into the SN or into the caudate nucleus or by (+)-3PPP (1.0 nml) microinjected into the n. accumbens or applied onto the prefrontal cortex. 3. Behavioural and ECoG sleep was also induced in rats after systemic administration of apomorphine (263 nmol kg-1, i.p.). 4. The behavioural and ECoG spectrum power effects of apomorphine (1.0 nmol) bilaterally micro-infused into the VTA were prevented by a previous microinjection into the same site of (-)-sulpiride (9.8 nmol). Similarly, behavioural and ECoG effects evoked by (+)-3PPP (0.1 nmol) given bilaterally into the VTA, were completely antagonized by a previous injection into the same site of haloperidol (16 pmol given 10 min before). In contrast, pretreatment with SCH 23390 (50 pgkg-1, s.c.), a selective antagonist at dopamine Dl-receptors, was unable to antagonize the behavioural and ECoG spectrum power effects of ( +) 3PPP. 5. Soporific effects induced by systemic administration of apomorphine were antagonized by (-)- sulpiride (9.8 nmol) given bilaterally into the VTA 10min before, whereas, yohimbine (1.3 nmol), (an antagonist at alpha 2-adrenoceptors) bilaterally microinfused into the VTA, was ineffective in this respect. 6. The present experiments provide evidence suggesting that stimulation of dopamine D2 receptors located at the cell body level and/or the dendrites of dopaminergic neurones in the VTA may represent the mechanism through which apomorphine or (+) 3PPP exert their soporific effects in rats. PMID- 2974740 TI - Characteristics of cromakalim-induced relaxations in the smooth muscle cells of guinea-pig mesenteric artery and vein. AB - 1. The effects of cromakalim (BRL 34915) on the smooth muscle cells of guinea-pig mesenteric artery and vein were investigated with microelectrode and tension recording methods. 2. Cromakalim (greater than 10 microM) produced membrane hyperpolarization with an increase in ionic conductance. The hyperpolarization occurred to a greater extent and lasted longer in the vein than in the artery. 3. The hyperpolarization induced by cromakalim in mesenteric vein comprised two components, one of which was Mn sensitive. In mesenteric artery, the hyperpolarization was relatively insensitive to Mn. 4. From the current-voltage relationship measured from arterial smooth muscle membranes, the reversal potential of cromakalim was estimated to be -80 mV. The cromakalim-induced hyperpolarization was not modified in Na- or Cl-deficient solution. 5. In both mesenteric artery and vein, cromakalim relaxed tissues precontracted with high K with (below 40 mM) or without (above 40 mM) hyperpolarization of the membrane. 6. In the mesenteric artery, action potentials evoked by electrical stimulation ceased before the generation of hyperpolarization. 7. Cromakalim produced a cross desensitization with nicorandil on the evoked membrane hyperpolarization in mesenteric artery. 8. It is concluded that the relaxing actions of cromakalim result from the hyperpolarization which follows the opening of Ca-dependent K channels. The inhibition of a voltage-dependent Ca current may also be involved in this inhibitory effect. PMID- 2974743 TI - Fibromuscular dysplasia of the external iliac artery: treatment by percutaneous transluminal angioplasty. PMID- 2974744 TI - Percutaneous transluminal angioplasty and vascular surgery for lower limb ischaemia. PMID- 2974745 TI - Language functions in Huntington's disease. AB - A comprehensive language test battery (Aachen Aphasia Test) was administered to 45 patients in the early, middle or later stages of Huntington's disease (HD) and to 20 control subjects. In spontaneous speech, many HD patients exhibited a loss of conversational initiative. Dysarthria was a common finding. Reading skills were found to be impaired mainly as a consequence of dysarthria; some HD patients displayed visual dyslexia. In addition to the characteristic disturbances of writing skills due to the choreiform movement disorder, the writing of HD patients with advanced dementia indicated constructional dysgraphia, characterized by frequent omissions, perseverations and substitutions. HD patients exhibited no evidence of word-finding difficulty or other semantic deficits in spontaneous speech. There was, however, a marked impairment in visual confrontation naming, with a significant rise in naming error rate as the disease progressed in severity. In most instances, the inappropriate names referred to an object visually similar to the target object, suggesting that visual misperception is the major cause of the naming disorder in HD. Syntactical structure of spontaneous speech was typically reduced to short, simple sentence construction. Verbal stereotypes were only rarely encountered and occurred late in the course of the disease. Tests of language comprehension reflected the general degree of dementia. It is concluded that there are no primary language changes in HD. Instead, a variety of language impairments develop secondary to other neurological and neuropsychological changes. PMID- 2974746 TI - Dopamine D2 receptor stimulation inhibits inositol phosphate generating system in rat striatal slices. AB - Previous studies on the transduction mechanisms triggered by dopamine receptor stimulation have established that both D1 and D2 subtypes of dopamine receptors are linked to the adenylate cyclase system, the former in a stimulatory and the latter in an inhibitory manner. The present report provides the first evidence that stimulation of D2 receptors in rat brain tissue affects the turnover of polyphosphoinositides, as revealed by changes of the content of inositol phosphates. We found that the basal level of [3H]inositol trisphosphate, [3H]inositol bisphosphate and [3H]inositol monophosphate decreased following the stimulation of the D2 receptor. The rank order of potency was quinpirole (IC50 5 nM) greater than lisuride (IC50 8 nM) greater than RU 24213 (IC50 50 nM) greater than dopamine (IC50 200 nM). In contrast, selective D1 receptor stimulation by fenoldopam did not alter the inositol monophosphate, inositol bisphosphate and inositol trisphosphate content. The quinpirole effect was prevented by selective D2 antagonists, such as domperidone and L-sulpiride (both 5 microM) while it was unaffected by the selective D1 antagonist SCH 23390 (100 nM) and by the pharmacologically inactive D-isomer of sulpiride. Our data indicate that the activation of striatal D2 receptors leads to the inhibition of inositol phosphate production. PMID- 2974747 TI - Regional differences in reappearance of D2-dopamine receptors in the rat caudate putamen complex after irreversible inactivation. AB - The reappearance of D2-receptors in the striatum of the rat was studied by autoradiography after in vivo labeling with [3H]N-n-propylnorapomorphine ([3H]NPA) at various time intervals after the inactivation of dopamine receptors by intraperitoneal administration of N-ethoxycarbonyl-2-ethoxy-1,2 dihydroquinoline (EEDQ). Within two days after inactivation the labeling had decreased to 18% of controls. Thereafter, the label reappeared and after 8 days or more reached levels of 80% of that of untreated controls. Autography showed that 4 h after EEDQ treatment no preferential labeling of the striatum can be seen. Five days after EEDQ a slight difference in labeling density between the medial and lateral striatum was detected, whereas after 18 days a prominent lateromedial gradient in silver grain density was seen, resembling the gradient seen without EEDQ treatment. This silver grain gradient is not paralleled by the density of medium-sized neuronal cell bodies. This suggests a difference in synthesis rate of receptors either in other cells than the medium-sized neuron or, alternatively, in otherwise indistinguishable medium-sized neurons. Five days after EEDQ treatment, clusters of silver grains in the lateral striatum were seen. These clusters have a diameter of 150-400 microns and are separated from each other at 200-500 microns. Each cluster may represent newly synthesized receptors of a single neuron (e.g. cholinergic or somatostatinergic interneuron). PMID- 2974748 TI - Physiological implications of the alteration of 6-phosphofructo-1-kinase isozyme pools during brain development and aging. AB - The 6-phosphofructo-1-kinase (PFK) isozyme pools from brains of fetal, neonatal, young adult (3 months) and aged (30 months) rats were studied using chromatographic and immunological techniques. Also, the changing subunit composition of each isozyme pool was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis on 6% slab gels and by immunoblotting with subunit-specific antibodies. The total PFK activity increased over seven-fold during the 30 days following birth, and the L-type, M-type, and C-type subunits increased approximately 2-fold, 7-fold, and 24-fold, respectively. In the near term fetal brain and early neonatal brain, the L-type and M-type subunits were the predominant forms and were present in approximately equal amounts. During the second second week of postnatal brain maturation, the levels of the M-type and C type subunit began to significantly increase. Consequently, during postnatal development, the isozyme pools switched from L-M-rich forms to M-C-rich forms. In aged brain relative to the young adult (3 months) brain, the 20% loss of total activity was associated with 27% and 18% losses of the M-type and C-type subunits, respectively. Examination of the regulatory properties of the various PFK isozyme pools revealed that at the low concentration of fructose-6-P and high level of ATP which are thought to occur in vivo, fructose-2,6-P2 was required for measurable PFK activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974749 TI - Electrical properties of cultured dorsal root ganglion neurons from normal and trisomy 21 human fetal tissue. AB - Dorsal root ganglion (DRG) neurons, in 22 degrees C tissue culture containing nerve growth factor, taken from normal and trisomy 21 human fetal tissues, were subjected to current and voltage clamp measurements using a tight-seal whole-cell recording technique. Measurements were made between 1 and 2 weeks in culture, when the electrical properties of both neuron groups were shown to be constant and when mean values for passive electrical parameters did not differ significantly between groups. The duration of the action potential was significantly less in trisomic than in control neurons, and both depolarization and repolarization were accelerated. Tetraethylammonium (5 mM), which partially blocked outward currents, prolonged the rate of repolarization of the action potential in both neuron groups, and abolished the difference in the rate between the groups. Furthermore, the activation rate constants of two model-defined outward potassium currents were significantly higher in trisomic than in control neurons, suggesting that acceleration of repolarization of the action potential in trisomic neurons was due to shorter activation time-constants of outward potassium currents. PMID- 2974750 TI - Phenotypic expression of photoreceptor and endocrine cell properties by cultured pineal cells of the newborn rat. AB - Pineal glands of newborn rats were dissociated and maintained under cell culture conditions. The phenotypic expression of both photoreceptor and endocrine cell properties was investigated using immunohistochemical techniques (specific antibodies against opsin or serotonin). After one week in culture, a number of small round cells appeared on top of a sheet of flat epithelium. Among those cells, opsin-like immunoreactive cells were observed. These cells showed a neuron like morphology with neuritic processes and often formed rosettes. Immunoreactivity was found on the plasma membrane of both the soma and cell processes. Serotonin-like immunoreactive cells were also differentiated in culture with two different morphological types of cells being found. One type resembled cultured serotonin-containing amacrine cells of the retina, and the other type had a flat, polygonal shape similar to that of pinealocytes. Both types of immunoreactive cells possessed fine neuritic processes. These results indicated that cell culture of rat pineal gland cells allowed expression of some properties, such as opsin synthesis and neuron-like morphology with long neuritic processes, that were not expressed in the intact rat pineal gland. PMID- 2974751 TI - Perturbations of lens development in the transgenic mouse. AB - Interference with specific features of cell growth and differentiation in vivo offers rare insights into the complex mechanism of cell proliferation in the developing organism. We have inserted in the germ line of mice transgenes which direct dominant changes in cell development and proliferation to the lens, a tissue exquisitely suited for in vivo analysis. Lens-specific expression of SV40 T antigens results in malignant lens tumors or phakomas, that of a murine c-mos protooncogene in a characteristic defect of lens fiber differentiation. PMID- 2974752 TI - Adjunctive methods of examination of the urogenital tract. AB - Included in this article are descriptions of adjunctive methods of examination of the urogenital tract, including ultrasonography of the kidneys, urinary bladder, ovaries and uterus; endoscopy of the urethra, urinary bladder, and uterus; contrast radiography of the urinary tract; kidney biopsy; and laparoscopy of the abdomen, emphasizing examination of the mare's reproductive tract. PMID- 2974753 TI - Surgery of the penis and prepuce. AB - Treatments of penile and preputial injuries, anomalies, tumors, "summer sores," paraphimosis, phimosis, and priapism are presented. Surgical procedures described include preputiotomy, reefing, penile retraction, amputation, and relocation. Treatment of hemospermia by means of urethrotomy is discussed. PMID- 2974754 TI - Effect of maternal insulin deficiency on vitamin D metabolite concentrations in normoglycemic pregnant rats and their fetuses. AB - The effect on vitamin D metabolite concentrations of insulin deficiency, not accompanied by hyperglycemia, were investigated in pregnant rats and in their fetuses injected with 75 mg/kg BW streptozotocin (SZ). These concentrations were measured in maternal plasma and whole fetal body. In the insulinopenic mothers, the 25OHD concentration was found to rise compared with that of control pregnant rats (7.00 +/- 1.66 ng/ml, n = 16, versus control 4.50 +/- 1.60, n = 10, 0.001 less than P less than 0.01). The concentration of 1,25(OH)2D, which was previously found to decrease in pregnant rats that were both hypoinsulinic and hyperglycemic, was previously found to decrease in pregnant rats that were both hypoinsulinic and hyperglycemic, was not different in our control and insulinopenic rats (107.36 +/- 38.25 pg/ml, n = 11, versus control 122.90 +/- 18.20, n = 18.20, n = 8). In fetuses from our SZ-injected rats, the 24,25(OH)2D level diminished compared with the control level (2.12 +/- 0.70 ng/g, n = 11, versus control 5.23 +/- 0.95 ng/g, n = 13, P less than 0.001). The Ca/P ratio in fetal body also decreased (0.68 versus control 1.12). It is suggested that the placental metabolism is an important determinant or normal fetal growth. PMID- 2974756 TI - The ultrastructure and ATPase nature of polar membrane in Campylobacter jejuni. AB - Polar membrane in Campylobacter jejuni has been visualized on membrane vesicles. It was composed of doughnut-shaped particles 5-6 nm in diameter, with stalks, arranged in a hexagonal array. This structure was stabilized on the membrane by a high ionic strength buffer in the presence of 2-mercaptoethanol. Histochemical staining indicated localized ATPase activity at the poles of the cells. An ATPase with distinctive properties has been isolated and purified from this organism; it gives a specific activity of approximately 0.3 units/mg of protein. Electron microscopy showed doughnut-shaped particles 5-6 nm in diameter. Nondissociating and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme revealed, respectively, a single band with ATPase activity and a molecular weight of ca. 75,000 Da. The enzyme was cold labile and activity was abolished by trypsin. Dicyclohexylcarbodiimide inhibited the membrane-bound form of the enzyme, but did not inhibit the soluble form. Oligomycin had no inhibitory activity on either form of the enzyme. The enzyme specifically hydrolysed ATP, but other nucleotide substrates were not degraded. The enzyme was activated by Mg2+ and inhibited by Ca2+, whereas other ions had no effect on activity. Antibodies prepared to this enzyme bound to the polar regions of whole cells as shown by protein A - colloidal gold immunoelectron microscopy. The antibodies to this ATPase cross reacted (shown by Western blotting) with four proteins from a whole-cell extract of this organism, two proteins in Aquaspirillum serpens MW5, and three proteins from Escherichia coli K12. They did not cross-react with any proteins from Spirillum volutans, Methanococcus voltae, Vibrio cholerae, or rat liver mitochondria. Antibodies raised against the F1-ATPase of E. coli K12 cross reacted with six proteins in a whole-cell extract of this organism, and one protein species in each of the whole-cell extracts of V. cholera, A. serpens MW5, S. volutans, and rat liver mitochondria. These antibodies did not recognize any whole cell proteins from either C. jejuni or M. voltae. These results along with the ATPase activity localized by histochemical staining suggest that polar membrane is an assembly of ATPase molecules at the poles of the cell and that the ATPase isolated from C. jejuni is serologically and structurally unusual. PMID- 2974755 TI - [The effect of cytochalasin A on the composition of subcellular fractions of hyphae in the growth of Mucor mucedo L. I. Composition of the plasmalemma]. AB - The plasma membrane of young hyphae of Mucor mucedo L. growing in presence or absence of cytochalasine A was isolated by continuous density gradient centrifugation using Percoll at 10% or on discontinuous sucrose density gradient. Isolated membranes were characterized by enzymatic markers and cytochemical reactions, using electron microscopy. Lipid composition and protein content were determined. From the enzymatic point of view, the cytochalasine A induced a decrease (60%) in ATPase activity and with regard to the chemical composition of the membrane, a decrease in sterol content and in the sterol-phospholipid ratio as well as a decrease in protein content and an increase in the proportion of cysteine relative to other amino acids. PMID- 2974757 TI - The role of medroxyprogesterone acetate (MPA) in the treatment of renal adenocarcinoma. PMID- 2974758 TI - Recognition of oligosaccharide substrates by N-acetyl-glucosaminyltransferase-V. AB - Six analogs of the trisaccharide 8-methoxycarbonyloctyl 6-O-[2-O-(2-acetamido-2 deoxy-beta-D-glucopyranosyl)-alpha-D-mannopyrano syl] -beta-D-mannopyranoside (3), a previously reported acceptor for N-acetylglucosaminyltransferase-V (GnT-V) have been chemically synthesized and evaluated as GnT-V acceptors. Replacement of the beta-D-man rho-O(CH2)8COOMe "reducing end" of 3 by beta-D-Glc rho-O(CH2)7 CH3 gave octyl 6-O-[2-O-(2-acetamido-2-deoxy-beta-D-glucopyranosyl]-alpha-D- mannopyranosyl)-beta-D-glucopyranoside (5) whose activity was indistinguishable from that of 3. Removal of the 4-OH group of the beta-D-Glc residue in 5 had little effect on the activity, while the corresponding 4-O-methyl derivative was twice as active. Replacement of the C-6 pro-R hydrogen of the same residue by a methyl group gave the L-glycero-D-gluco derivative 8, whereas replacement of the corresponding pro-S hydrogen gave the D-glycero-D-gluco compound 9. Trisaccharide 8, whose rotameric distribution about the C-5-C-6 bond is sterically biased towards the gg conformation was less than half as active as 5 as a GnT-V acceptor, whereas 9, which is biased towards the gt conformation, was more than twice as active. These results provide evidence for the conformational control of oligosaccharide biosynthesis. PMID- 2974759 TI - Neisseria perflava amylosucrase: characterization of its product polysaccharide and a study of its inhibition by sucrose derivatives. AB - Neisseria perflava amylosucrase forms from sucrose a polysaccharide very similar to glycogen, except that a larger proportion of its D-glucosyl residues are in short branches. Iodine staining of samples taken during polysaccharide formation indicate that the initial product is less branched than that formed at longer times. This glycogen-like polysaccharide has an estimated molecular mass range of 1 MD to 20 MD. Sucrose derivatives modified at C-3 (3-deoxysucrose and alpha-D allopyranosyl beta-D-fructofuranoside), C-6 (6-deoxysucrose and 6-deoxy-6 fluorosucrose), and both C-4 and C-6 (4,6-dideoxysucrose) were tested as inhibitors of amylosucrase. Derivatives modified at C-6 were potent competitive inhibitors, with Ki values of 6.2 +/- 0.3 mM (6-deoxysucrose) and 0.50 +/- 0.06 mM (6-deoxy-6-fluorosucrose). The KM value of sucrose is 26.5 +/- 4.6 mM. Sucrose derivatives modified at C-3 were not significantly inhibitory over the concentration range tested. 4,6-Dideoxysucrose gave an unusual, non-competitive inhibition, in that, increasing its concentration did not produce a commensurate increase in the level of inhibition, which instead appeared to approach a limit. None of these sucrose derivatives was a substrate for amylosucrase, nor were they glycosyl donors to maltotriose. PMID- 2974760 TI - Structure and mass of mammalian respiratory ciliary outer arm 19S dynein. AB - Mammalian respiratory ciliary outer arm dyneins isolated as the major ATPase peak migrating at 19S on sucrose density gradients were examined by transmission electron microscopy of negatively stained samples and scanning transmission electron microscopy of unstained samples. The predominant discrete particle structure observed was composed of two globular heads apparently connected by amorphous or indistinct material. The heads were either circular or slightly elliptical of mean 13 +/- 1 X 10 +/- 2 nm dimensions. The mass of this structure averaged 1.22 +/- 0.34 million daltons with the individual globular heads averaging 310 +/- 77 kilodaltons (kD). Negative staining revealed that one or both of the globular heads often contained a central accumulation of stain measuring 2.5 +/- 1 nm across. A second type of structure, appearing with lesser frequency in the 19S fraction than in the unfractionated dynein preparation loaded onto the sucrose gradient, was a single globular head of 13 +/- 1 X 10 +/- 2 nm often with 2 +/- 1 nm centrally accumulated stain and with or without an appendage. This one-headed particle thus resembled one-half of the two-headed particle. Mass measurements were lower, however, for isolated, single globular heads, averaging 220 +/- 111 kD. A third type of particle observed was a ring like structure with 4 +/- 1 nm centrally accumulated stain and without appendages. The ring structure was slightly larger in diameter, 14 +/- 1 nm, and had a greater peripheral accumulation of negative stain than either of the one- or two-headed particles, suggesting that it was not derived therefrom.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974761 TI - Force production and microtubule-coupled cell movement. Conference on Cell Motility. August 20-25, 1988, Bolton Valley, Vermont. Abstracts. PMID- 2974762 TI - [The care of patients undergoing gynecologic laparoscopy examination]. PMID- 2974764 TI - White nails in AIDS/ARC due to Trichophyton rubrum infection. PMID- 2974765 TI - Presence of an IL-3-producing suppressor T cell resistant to cyclosporin A in the peripheral blood of patients with systemic lupus erythematosus. AB - We studied the effects of cyclosporin A (CYA) on the production of various lymphokines and on the suppressor functions of peripheral blood mononuclear cells (MNC) of ten untreated SLE patients. CYA was found to inhibit the production of IL-2 and of B cell stimulating factor (BSF) by both normal and SLE MNC but it did not alter the LPS-driven production of IL-1 by either of them. However, it did abrogate the spontaneous release of IL-1 by SLE monocytes. CYA strongly inhibited the production of IL-3-like activity by normal T cells, an effect noticed only on cells from 4 SLE patients, three of whom were in remission. Addition of CYA significantly increased suppressor cell function by cells from all SLE patients. T cell clones could be obtained from two of them by using CYA-conditioned medium containing IL-3. These clones were found to have strong suppressor capacity. PMID- 2974763 TI - Helper strategy in tumor immunology: expansion of helper lymphocytes and utilization of helper lymphokines for experimental and clinical immunotherapy. AB - Two main kinds of immune strategy are possible against neoplasia. The first potentiates a selected effector arm. In vitro culture with exogenous interleukin 2 (IL-2) increases the activity of natural killer cells and leads to the expansion of T cytotoxic lymphocytes. Systemic reinfusion of both of these cells with high doses of IL-2 mediates the regression of a variety of murine and human tumors. In an alternative strategy, a few regulatory lymphocytes turn on immune reactivity by triggering a cascade of interconnected effector functions. The efficacy of this strategy rests on the repertoire of effector mechanisms moved to action. An effective immunoregulatory maneuver is the addition of helper determinants on the surface of tumor cells. Its power can be further increased by the pre-induction of helper T lymphocytes specific to the helper determinants. This approach can be achieved in mice by coupling muramyl dipeptides to tumor cells, along with eliciting T lymphocytes specifically reactive to Bacillus Calmette-Guerin. Noncytotoxic T helper lymphocytes produce factors which recruit nonspecific (macrophages) as well as specific (cytolytic T lymphocytes) anti tumor attacking cells. In this way protection can be afforded against primary tumors and metastases, as well as leukemia cells. As the activity of helper lymphocytes rests mostly on lymphokine release, the use of molecularly defined lymphokines mimicking T-helper functions has also been attempted. In a few experimental models, the association of low doses of IL-2 with non-reactive lymphocytes from tumor-bearing mice promotes an effective anti-tumor reaction in the host. Moreover, the combination of distinct lymphokines can also build a molecularly defined helper system able to activate in sequence non-specific and specific anti-tumor reactions in vivo. Trials intended to evaluate the clinical impact of these helper approaches in the management of human tumors are being started or are already under way. PMID- 2974766 TI - Analysis of negatively charged dye-binding antibodies reactive with double stranded DNA and heparan sulfate in serum from patients with rheumatic diseases. AB - Antibodies to double-stranded (ds) DNA are characteristically present in serum from patients with systemic lupus erythematosus (SLE). Recently, anti-dsDNA antibodies have been shown to have the capacity to react with a diversity of molecules with repeating negative charges. Using the anionic dye Cibacron blue F3GA, bound to crosslinked agarose, we analysed the nature of antibodies capable of reacting with this dye in serum samples from patients with various rheumatic diseases. The dye-antibody complex could easily be split by eluting with solutions of increasing ionic strength, suggesting that the interaction is ionic in nature. Pepsin-digested F(ab')2 antibodies retained the capacity to bind Cibacron blue, confirming that the binding occurred via antigen-binding sites on the antibody molecule. The eluates obtained from dye-ligand chromatography of active SLE sera contained antibodies to both dsDNA and heparan sulfate, while those of sera from patients with other non-SLE rheumatic diseases contained antibodies only against heparan sulfate. Furthermore, the dye-ligand eluates of sera from patients with active SLE and other non-SLE rheumatic diseases were found to contain increased amounts of IgG. In one patient with SLE, levels of antibodies to dsDNA and heparan sulfate, and the amounts of total IgG in dye ligand eluates, were shown to be correlated with disease activity. PMID- 2974767 TI - Anticardiolipin antibodies in systemic sclerosis: immunological and clinical associations. AB - Anticardiolipin antibodies of IgG/IgM class were detected in seven of 28 patients with systemic sclerosis including five of 16 patients severely affected by extensive visceral disease. This severely affected sub-group also showed significant elevations of plasma levels of von Willebrand factor antigen in 10 cases and serum C1q binding activity in seven cases respectively. This triple association raises the possibility that multiple immunological mechanisms are involved in the pathogenesis of systemic sclerosis and its vascular lesions. PMID- 2974768 TI - C3b receptor (CR1) on phagocytic cells from SLE patients: analysis of the defect and familial study. AB - In vitro CR1-dependent phagocytosis of C3b-coated erythrocytes, by monocytes and PMN, was found to be significantly decreased in SLE patients. This was in many cases related to a specific defect of CR1 receptors, since the FcR-ingestion of EIgG was normal. On the other hand, CR1 levels of PMN stimulated by FMLP were also found to be decreased in SLE patients, while both the expression of circulating PMN (cells isolated at 4 degrees C) and the total cellular CR1 content were normal. On the basis of the available data, we propose that the impaired phagocytosis is due to a functional defect of CR1 or a defective anchorage of the receptor to the plasma membrane, possibly related to the decreased capacity of CR1 to be up-regulated by FMLP. To study the importance of the genetic background in the CR1 abnormalities, the families of 22 young SLE patients, in which the onset of the disease had occurred before the age of 15, were studied. The expression of CR1 on erythrocytes, and the total CR1 content of PMN, in parents and siblings of these patients did not differ significantly from normal controls. By contrast, the ingestion of EIgGC3b by PMN from healthy relatives of these patients was decreased (65% of the normal mean of PI), while EIgG phagocytosis was normal. A relation between this CR1 functional defect and the familial occurrence of autoimmune disorders is therefore possible. PMID- 2974770 TI - Electrocardiographic diagnosis of left ventricular hypertrophy: influence of body build. AB - 1. The sensitivity and specificity of four sets of electrocardiographic criteria for detection of left ventricular hypertrophy were evaluated in an echocardiographic study of 100 hypertensive patients. 2. All criteria gave reasonable specificity (87-94%) but poor sensitivity (39-52%). 3. When non-obese and obese patients were studied separately, criteria based on chest lead voltages were more sensitive than limb lead criteria for detection of left ventricular hypertrophy in non-obese subjects; however, the reverse was true in obese hypertensive patients, where criteria based on limb lead voltages were more sensitive than chest lead voltage criteria. 4. These data suggest that stratification of subjects by body build might improve the diagnostic performance of the electrocardiogram for detection of left ventricular hypertrophy. PMID- 2974769 TI - Domperidone, a DA2-specific dopamine antagonist, has no effect on the renal or haemodynamic response to atrial natriuretic peptide in man. AB - 1. Animal experiments have suggested that the renal effects of atrial natriuretic peptide (ANP) are dependent on dopaminergic activation, predominantly of the DA1 receptor. In man, there is evidence of dependence on the DA2-receptor for the natriuresis produced by central blood volume expansion. 2. Six normal volunteers underwent infusions of alpha-human ANP preceded by domperidone (a DA2-antagonist) or placebo. Eight volunteers underwent a 3 h period of 10 degrees head-down tilt also preceded by domperidone or placebo. 3. Both the ANP infusion and head-down tilt produced a significant diuresis and natriuresis, neither of which was antagonized by the presence of domperidone. 4. The ANP infusion significantly reduced diastolic blood pressure and produced significant increases in the Doppler-measured aortic blood velocity variables of peak velocity and maximal acceleration. Domperidone had an independent effect of increasing blood pressure but did not appear to have a specific interaction with the haemodynamic effects of ANP. 5. Head-down tilt reduced mean arterial pressure and heart rate and increased maximal acceleration. Again, an independent effect of domperidone was seen on blood pressure. Heart rate and maximal acceleration showed similar changes in the presence of domperidone. 6. Domperidone does not antagonize the renal or haemodynamic effects of ANP and if dopaminergic activation is necessary for the renal action of ANP it is unlikely to be mediated by the DA2-receptor. PMID- 2974771 TI - Magnesium aspartate hydrochloride attenuates monocrotaline-induced pulmonary artery hypertension in rats. AB - 1. The effect of oral magnesium aspartate hydrochloride on monocrotaline (MCT) induced pulmonary arterial hypertension was evaluated in rats. 2. A single subcutaneous injection of MCT, a pyrrolizidine alkaloid of plant origin, induces significant morphological changes in pulmonary vessels, pulmonary arterial hypertension and right ventricular hypertrophy in rats by 3 weeks. 3. Two groups of rats (Mg2+ control and Mg2+ + MCT) were started on oral Mg2+ (15.4 g/l magnesium aspartate hydrochloride dissolved in deionized water) 2 weeks before the MCT injection. The rest were given deionized water. At the start of the experiment, the control groups (deionized water and Mg2+) were given normal saline subcutaneously; the other groups (deionized water and Mg2+) were given MCT (60 mg/kg) subcutaneously. 4. Pulmonary artery pressure, right ventricular hypertrophy, lung pathology, organ weights and serum electrolytes were assessed 3 weeks after a single subcutaneous injection of MCT. Seventy-five per cent of the rats treated with MCT and oral Mg2+ (12 out of 16) showed significant reduction in pulmonary arterial hypertension, arterial pathology and right ventricular hypertrophy. 5. Our data indicate that Mg2+ attenuates experimentally induced pulmonary hypertension, possibly either by modulating the intracellular Ca2+ level and/or by directly affecting the pulmonary endothelial cell-smooth muscle cell complex involved in metabolism and maintenance of pulmonary vascular resistance. PMID- 2974772 TI - Clinical strategies for obesity management. PMID- 2974773 TI - Diabetes and hypoglycaemia: medico-legal aspects of criminal responsibility. PMID- 2974774 TI - Collecting urine for microalbumin assay. PMID- 2974775 TI - A strategy for selection of elderly type 2 diabetic patients for insulin therapy, and a comparison of two insulin preparations. AB - Sixty-six patients with secondary failure to oral hypoglycaemic therapy were assessed in hospital, and those whose fasting blood glucose concentration was greater than or equal to 10.0 mmol l-1 were treated with insulin once a day for 6 months. Only 22 patients fulfilled this criterion, and they were randomly allocated to a daily injection of either Humulin-Zn (12 patients) or Neulente insulin (10 patients). The remaining patients were considered to be noncompliant with therapy. At the end of 6 months' insulin therapy a significant (p less than 0.05) improvement occurred in HbA1c from a median (range) of 13.2(9.8-16.4)% and 13.1(10.5-16.2)% to 10.6(8-14.2)% and 11.2(8.7-13.5)% in patients given Humulin Zn and Neulente, respectively. However, there were 46 episodes of hypoglycaemia reported by patients who received Humulin-Zn, 36 of which occurred between 0300 and 0600 h. Only four episodes were reported by the Neulente-treated group. In addition, six patients treated with Humulin-Zn (but only one patient on Neulente insulin) required the addition of short-acting insulin. The patients not treated with insulin showed a significant (p less than 0.05) improvement in blood glucose control 2 months after in-patient assessment but this improvement was not sustained to 6 months. PMID- 2974777 TI - The fluorescence of serum proteins in diabetic patients with and without retinopathy. AB - Non-enzymatic glycosylation of long-lived proteins results in a characteristic fluorescence, known as protein 'browning'. The degree of fluorescence of skin collagen correlates with retinopathy in diabetes. We have therefore measured the fluorescence of serum albumin and IgG by a sensitive HPLC technique in 69 diabetic patients, 38 with retinopathy and 31 without complications, and in 26 age-matched controls. The fluorescence of the IgG fraction, calculated as the ratio of fluorescence (excitation 360 nm, emission 454 nm) to optical density (280 nm), was elevated in diabetic patients with retinopathy (1.5 +/- 0.5) compared with those without retinopathy (1.1 +/- 0.2, p less than 0.005) and control subjects (1.1 +/- 0.2, p less than 0.005). The fluorescence ratio for the albumin fraction was increased in all diabetic patients (11.2 +/- 2.7) compared with control subjects (8.9 +/- 1.5, p less than 0.001). There was no significant difference in the fluorescence ratios of albumin in those with and without retinopathy. The fluorescence of serum albumin and IgG were not significantly correlated with serum protein glycosylation measured by the fructosamine method. PMID- 2974776 TI - Aldose reductase inhibition in diabetic neuropathy: clinical and neurophysiological studies of one year's treatment with sorbinil. AB - In a double-blind placebo-controlled trial the effect of Sorbinil (250 mg daily) on diabetic neuropathy was examined. After a 2-month run-in placebo period (with three major assessments) 21 patients were randomized to Sorbinil and 10 to placebo, and all were studied for a further 12 months with neurophysiological measurements at 3-month intervals of nerve conduction velocity in multiple nerves, autonomic function tests, vibration thresholds as well as clinical examination and an extensive self-assessment of symptoms. Two subjects on Sorbinil treatment developed a hypersensitivity reaction and were withdrawn. Metabolic control and severity of neuropathy was not significantly different between groups. There were no changes in symptoms as judged by self-assessment scores. No patient entered the trial with neuropathic ulcers but ulceration developed in 4 patients during Sorbinil treatment and in 1 of the placebo group. No beneficial effect of Sorbinil was demonstrated on either the clinical manifestation or on the neurophysiological measurements made in these neuropathic diabetic patients over 12 months of treatment. PMID- 2974778 TI - Group education for obese patients with type 2 diabetes: greater success at less cost. AB - It has been suggested that much effort expended in teaching diabetic diets is ineffective and wasteful. We have tested a different system by randomly allocating 75 newly diagnosed obese Type 2 diabetic patients to usual 'unstructured' clinic care or to group education by diabetes specialist nurses and a dietitian. Patients allocated to group education attended five 90-min group sessions during the first 6 months. Six months after diagnosis they had lost more weight (median (95% Cl), 7 (5.5-9) vs 2(1-5)kg, p less than 0.002) and were better controlled (HbA1:7.5 (7.0-8.1) vs 9.5 (8.7-10.4)%, p less than 0.001) than those randomized to the usual clinic system. At 1 year (after no further visits) the difference in weight loss was less (5.5 (4-6.5) vs 3 (2-4) kg, p less than 0.05) and diabetic control was similar (HbA1:9.0(8.2-9.8) vs 9.9(8.9-10.9)%. At 1 year only 14(39%) of the education group and 9(23%) of those attending the clinic had a fasting blood glucose less than 7.0 mmol l-1. PMID- 2974780 TI - Reduced hyperaemic response under the diabetic neuropathic foot. AB - Diabetic neuropathic ulcers typically occur at high pressure sites. Microvascular blood flow has been assessed on the plantar surface of the foot in three matched groups each of 12 subjects free from macrovascular disease: (a) patients with diabetic neuropathy with abnormal foot pressures and previous neuropathic ulceration; (b) non-neuropathic diabetic patients; (c) non-diabetic control subjects. Resting flow was measured at the highest pressure point under the metatarsal heads (defined by pedobarograph) using laser doppler flowmetry, and the hyperaemic response was assessed at the same site following 3 min standing. Peak flow was significantly reduced in neuropathic patients (2.3 +/- 1.4 (SD) volts) compared with control subjects (4.0 +/- 2.0 volts; p less than 0.03). The time for blood flow return to baseline was significantly prolonged in neuropaths (159 +/- 72 s) compared with normal subjects (93 +/- 18 s; p less than 0.01), with a significant delay also seen in non-neuropaths (151 +/- 38 s; p less than 0.0001 compared with normal). PMID- 2974779 TI - The abbreviated glucose tolerance test in screening for diabetes: the Islington Diabetes Survey. AB - The World Health Organization has recommended a single 2-h post-glucose load blood glucose level as a screening test for diabetes mellitus in epidemiological surveys. We have assessed its characteristics, when compared with a full supervised glucose tolerance test (OGTT), in estimating prevalence, and in diagnosing diabetes in the individual patient. A stratified sample of 223 of 1040 subjects who had participated in a diabetic survey that utilized a single capillary 2-h blood glucose estimation as a screening test were recalled for formal glucose tolerance testing. The numbers of subjects with diabetes at screening and at recall were similar (14/212, 6.6%; 13/216, 6.0%) but only 9 subjects were so classified on both occasions. Thirty-five subjects (16.5%) were suspected of having impaired glucose tolerance (IGT) at screening, and 52 (24.1%) at recall. There was substantial reclassification from screening IGT, with 3/35 worsening to diabetes, and 10/35 returning to normal. Capillary 2-h glucose levels gave an accurate assessment of the prevalence of diabetes but underestimated that of IGT. On the full OGTT, little difference in classification was found when the values of fasting and 1-h blood glucose were used in addition to those of the 2-h blood glucose used alone. The 2-h glucose had a within subject coefficient of variation of 32.4% which produced substantial reclassification of subjects with levels close to the diagnostic levels for diabetes, and this implies that such individuals should not be classified as having diabetes on the basis of a single glucose tolerance test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974781 TI - NPH insulin administration by means of a pen injector. AB - In an open randomized cross-over study 50 patients with insulin-dependent diabetes were allocated to 3 months of treatment with NPH insulin either by means of a pen injector (Insuject-X) or by conventional syringes. The needle of the NPH pen injector was removed immediately after use to avoid possible leakage of solvent. Ambulatory control was assessed every 6 weeks, including blood sampling (HbA1c and insulin antibodies) and recording of hypoglycaemia. NPH insulin containers were collected for insulin potency measurement by HPLC. A seven-point blood glucose profile was performed fortnightly by means of home blood glucose monitoring. At the end of the 6 months a questionnaire was completed. No significant changes occurred in HbA1c (difference 0.1 +/- 0.7 (SD)%), blood glucose profile, or the incidence of hypoglycaemic episodes on the two regimens. The concentration of NPH insulin in the containers remained constant. All but two of the patients preferred to continue to use the pen injector. This NPH pen injector is a reliable and efficacious tool which may also prove more convenient for the patients. PMID- 2974782 TI - Diabetes care in a UK health region: activity, facilities and costs. AB - A survey of district facilities and staffing for diabetes care in the South East Thames Region of England (population 3.58 million) is reported, and compared with the recommendations of the Royal College of Physicians and British Diabetic Association. Hospital Activity Analysis for diabetic patients in the Region for the year 1985 is also reported and costs assessed. Compared with recommended staffing levels there was a deficit of 12.8 Consultant Physicians with a weekly deficit of 109 sessions, and of 49.6 whole time equivalent Specialist Nurses. There were 11,857 admissions of diabetic patients, 4185 with diabetes as a primary cause. Seventy-six per cent of these were for diabetes without mention of complications and were therefore theoretically and potentially preventable. Total cost of diabetes care was estimated to be pounds 21.6 million p.a. for the Region of pounds 1.44 million per District Health Authority. We suggest it is time every Regional and District Health Authority gave some priority to diabetes care and established formal care strategies. To provide adequate staffing and facilities would improve standards of care, reduce morbidity and hospital admission rates, and be cost effective. PMID- 2974783 TI - Optimizing blood glucose control in type 2 diabetes: an approach based on fasting blood glucose measurements. AB - A structured approach to the management of Type 2 diabetes, aiming to reduce fasting blood glucose levels to near-normal, can provide effective blood glucose control with minimal risk of hypoglycaemia and in a manner acceptable to most patients. When the fasting blood glucose value is maintained in the region 4-6 mmol l-1, protein glycosylation and plasma triglyceride values usually become near-normal and this may help to prevent the development of long-term diabetic complications. We propose a simple management strategy, based on 3-monthly fasting blood glucose determination, which uses not more than two therapeutic agents at any one time. If diet and maximal oral therapy fail to keep fasting blood glucose levels below 6 mmol l-1 then the addition of a basal insulin supplement, e.g. from a once daily injection of ultralente insulin, can restore near-normal fasting blood glucose levels without the need for full insulin replacement therapy. In older patients, where long-term prevention of diabetic complications is not such an immediate priority, less strict blood glucose control may be reasonable, aiming to keep the fasting blood glucose values below 10 mmol l-1 in order to prevent symptoms secondary to glycosuria. Patients can be seen at a monthly general practice morning diabetic mini-clinic or with the aid of a nurse visiting elderly patients at home. PMID- 2974784 TI - Diabetes mellitus and Ramadan fasting: the need for a critical appraisal. PMID- 2974785 TI - An indwelling subcutaneous FEP cannula for intermittent insulin injection: patient experience and effect on diabetic control. AB - The acceptability and efficacy of injecting insulin through a subcutaneous FEP cannula (to reduce the frequency of needle pricks) compared with conventional multiple injection therapy was examined in a cross-over study. Thirty-two insulin dependent diabetic patients injected through the cannula for 10 weeks using a pen injector, followed by 10 weeks using the injector alone, or vice versa. Rapid acting insulin was given before meals and intermediate-acting insulin at bedtime. Blood glucose control was not affected by cannula use (glycosylated haemoglobin: cannula, 8.6 +/- 0.3%; no cannula, 8.6 +/- 0.3%). Twenty-two of the 30 patients completing the study preferred to use the cannula and 21 requested to continue using it. There were no complications associated with its use. PMID- 2974786 TI - Interval between injections and meals. PMID- 2974787 TI - Exchangeable sodium in diabetes. PMID- 2974790 TI - Blood glucose responses to sweetcorn and potato meals. PMID- 2974789 TI - A hamburger as a snack? PMID- 2974788 TI - Fructosamine. PMID- 2974792 TI - Atrial natriuretic peptide and other vasoactive hormones in volume regulation. PMID- 2974793 TI - Role of atrial natriuretic peptide in the postnatal adaptation of term and preterm infants. PMID- 2974791 TI - Effects of cyproterone acetate plus ethinylestradiol low dose on plasma androgens and lipids in mildly hirsute or acneic young women. AB - The purpose of this study was to compare, by a randomized double-blind study, involving 30 women with either acne or mild hirsutism, the effects on plasma androgens and lipoproteins, of two hormonal preparations used in acne and ensuring reliable contraception, namely DianeR (cyproterone acetate 2 mg and ethinylestradiol 50 micrograms per pill - D-50) and Diane 35 (cyproterone acetate 2 mg and ethinylestradiol 35 micrograms per pill - D-35). Both drugs, used over a 6-month period (treatment cycle of 21 days with a pill-free interval of 7 days), induced an impressive decrease of free testosterone levels and of 5 alpha androstane-3 alpha, 17 beta-diol-glucuronide, a parameter of peripheral androgen formation; also dehydroepiandrosterone sulfate levels decreased significantly. Except for an increase of both total T and dihydrotestosterone and as well as for a more important increase in SHBG levels during treatment with D-50, consequences of its more pronounced estrogen dominance in comparison to D-35, the effects of both drugs on androgen levels were similar. The effects on lipids were characterized by a non-significant increase of total cholesterol and a significant increase of HDL-C, HDL2-C and Apo-Al lipoproteins, which were somewhat more pronounced during D-50 treatment. Triglycerides increased significantly during D-50 but hardly during D-35 treatment, whereas HDL-C/LDL-C increased similarly during treatment with either drug. It is concluded that, being as efficient as D-50 in treatment of acne and hirsutism and taking into account its reliable contraceptive effects, without unfavourable effects, as far as atherogenesis is concerned, on plasma lipids, D-35 is to be preferred over D 50. PMID- 2974794 TI - Release of human natriuretic peptide in kidney transplantation. PMID- 2974795 TI - Atrial natriuretic peptide and other vasoactive hormones during treatment of severe diabetic ketoacidosis in children. PMID- 2974796 TI - Isotretinoin revisited. AB - Isotretinoin (13-cis-retinoic acid) remains the drug of choice for treatment of severe, recalcitrant nodulocystic acne that is unresponsive to conventional therapy, including oral antibiotics. The drug has been shown to produce dramatic clearing of lesions and prolonged remissions. The length of remission may be dependent on both the dosage used and the duration of therapy. Since fetal malformations have been observed in infants born of mothers taking isotretinoin during pregnancy, it is mandatory to prevent women who are pregnant or may become pregnant during treatment from taking isotretinoin. Because the half-life of isotretinoin is ten to twenty hours, and because it is eliminated from the body rapidly enough, women may become pregnant one month after discontinuation of therapy without an increased risk of birth defects. Common side effects of the drug include mucocutaneous reactions, serum lipid alterations, eye irritation, and myalgias. Less commonly observed are hyperostoses and exuberant granulation tissue, and rarely, pseudotumor cerebri. Recommendations for substantially reducing or eliminating these effects are made. PMID- 2974797 TI - Researchers cull AIDS information. PMID- 2974798 TI - Plasma atrial natriuretic peptide in states of altered thyroid function. AB - To examine a possible role of atrial natriuretic peptide (ANP) in water and electrolyte disturbances associated with thyroid disorders, plasma ANP levels were studied in patients with hyper- and hypothyroidism. In 5 of the 21 hyperthyroid patients, including two patients with atrial fibrillation and two patients with mild cardiomegaly, the plasma ANP concentration was increased when compared to normal subjects. After treatment with methimazole or propylthiouracil, the plasma ANP concentration fell to normal in 4 patients, while it remained high in one patient who had persistent atrial fibrillation. No significant correlation was found between plasma ANP and the heart rate in untreated hyperthyroid patients. Plasma ANP was within the normal range in all 8 patients with hypothyroidism. During treatment with T4, the plasma ANP concentration increased in 6 of the 7 patients. Chest X-ray films and ultrasonic echocardiography demonstrated pericardial effusion in 4 of these patients before therapy. A weak but significant correlation was found between the plasma ANP and T4 concentration, and between plasma ANP and free T4 in hyper- and hypothyroid patients before and after treatment. These results indicate that abnormalities in ANP dynamics in thyroid disorders may probably be caused by hemodynamic changes resulting from a thyroid hormone excess or deficiency. PMID- 2974799 TI - Heat shock but not other stress inducers leads to the disruption of a sub-set of snRNPs and inhibition of in vitro splicing in HeLa cells. AB - Splicing of pre-mRNA in HeLa cells exposed to various stress response inducers has been investigated. In vivo, intron-containing transcripts of the hsp27 gene accumulate in cells stressed by heat or sodium arsenite. In vitro analysis, however, reveals a differential effect of stress on splicing: nuclear extracts from cells exposed to a severe heat shock are incapable of splicing an exogenously supplied substrate while splicing is not perturbed in extracts treated with sodium arsenite, the amino acid analog canavinine or ethanol. Pretreatment of cells with a mild heat shock prior to a severe heat shock protects the splicing apparatus and allows splicing to proceed unimpeded. Analyses of the splicing defect in extracts from heat-shocked cells show that the inhibition of splicing cannot be accounted for by changes in the major RNA and protein components of small nuclear ribonucleoprotein particles (snRNPs) or in a previously described heat-labile factor that is essential for in vitro splicing. Fractionation of small nuclear ribonucleoprotein particles from heat-shock extracts by native polyacrylamide gel electrophoresis reveals dramatic changes in certain particles, most noticeably in a U4/U5/U6 snRNP complex and the U2 snRNP. Alterations in these particles are accompanied by the assembly of labeled pre mRNA transcript into aberrant splicing complexes that differ from those formed in normal extracts. PMID- 2974800 TI - Cells resistant to interferon are defective in activation of a promoter-binding factor. AB - Human cultured cell lines deficient in their ability to respond to type I interferon (IFN) fail to interrupt cellular proliferation or to induce an antiviral state following exposure to IFN alpha. Comparison of non-responsive Daudi and HeLa cell lines with IFN-responsive partner cell lines and examination of non-responsive Raji cells showed that the defective cell lines expressed type I IFN receptors of typical number and affinity and bound IFN equivalently compared to the normal cells. However, transcriptional induction of interferon stimulated genes (ISGs) was greatly reduced and delayed in these cell lines, leading to reduced accumulation of ISG mRNA. Furthermore, the rapid activation of IFN-stimulated promoter binding factors whose appearance correlates with ISG transcriptional induction, did not occur in non-responsive cells. Thus, the primary defect of these cells leading to an impaired physiological response to IFN appears to be an inability to activate promoter-binding factors necessary to trigger ISG transcription, an obligate early step in antiviral and antiproliferative physiology. PMID- 2974801 TI - Isolation and characterization of unusual gin mutants. AB - Site-specific inversion of the G segment in phage Mu DNA is promoted by two proteins, the DNA invertase Gin and the host factor FIS. Recombination occurs if the recombination sites (IR) are arranged as inverted repeats and a recombinational enhancer sequence is present in cis. Intermolecular reactions as well as deletions between direct repeats of the IRs rarely occur. Making use of a fis- mutant of Escherichia coli we have devised a scheme to isolate gin mutants that have a FIS independent phenotype. This mutant phenotype is caused by single amino acid changes at five different positions of gin. The mutant proteins display a whole set of new properties in vivo: they promote inversions, deletions and intermolecular recombination in an enhancer- and FIS-independent manner. The mutants differ in recombination activity. The most active mutant protein was analysed in vitro. The loss of site orientation specificity was accompanied with the ability to recombine even linear substrates. We discuss these results in connection with the role of the enhancer and FIS protein in the wild-type situation. PMID- 2974802 TI - Visualization of reversible macromolecular reactions in an analytical ultracentrifuge. AB - It has been shown that the saw-like disturbances of sedimentation observed in an analytical ultracentrifuge are not caused by convective disturbances of the solution but result from a special type of intermolecular reaction of reversible association/dissociation. A qualitative theory of saw-like anomalies has been suggested and the sedimentation and kinetic conditions of their origin have been indicated. Such reactions are a frequent occurrence in the serum of patients affected with rheumatic diseases and acute myocardial infarction. Experimental data indicate the involvement of immunoglobulins (viz., low-affinity antibodies) which form reversible immune complexes. Saw-like sedimentation patterns, especially those of the schlieren type, are a direct testimony to reversible association/dissociation reactions in macromolecular solutions, whereas other experimental methods provide only oblique evidence. PMID- 2974803 TI - Domain preference in iron removal from human transferrin by the bacterial siderophores aerobactin and enterochelin. AB - The ability of the siderophores aerobactin and enterochelin to remove iron from transferrin is reported. Aerobactin removes iron from both high-affinity sites on the transferrin molecule, but shows a marked preference for the C-terminal site. This preference is different to that of many iron chelators. Enterochelin removes iron perferentially from the N-terminal site. No evidence for synergism between aerobactin and bidentate ligands could be detected. PMID- 2974804 TI - Distances between functional sites in cardiac sarcoplasmic reticulum (Ca2+ +Mg2+) ATPase. Inter-lanthanide energy transfer. AB - The high-affinity Ca2+-binding sites of cardiac sarcoplasmic reticulum (Ca2+ +Mg2+)-ATPase have been probed using trivalent lanthanide ions. Non-radiative energy-transfer studies, using luminescent probe Eu3+ as a donor and Nd3+ or Pr3+ as acceptor, were carried out to estimate the distance between two high-affinity Ca2+-binding/transport sites. Eu3+ was excited directly with pulsed laser light and the energy-transfer efficiency to Nd3+ or Pr3+ was measured, under the conditions in which most donor-acceptor pairs occupied the high-affinity Ca2+ sites. The distance between two high-affinity Ca2+ sites is about 0.89 nm. In the presence of ATP the distance between the high-affinity sites is about 0.855 nm, whereas in the presence of adenosine 5'-[beta, gamma-methylene]triphosphate or adenosine 5'-[beta, gamma-imino]triphosphate the distance is about 0.895 nm. To estimate the distance between the high-affinity Ca2+ sites and ATP binding/hydrolytic site, we have measured the energy-transfer efficiency between Eu3+ and Cr3+-ATP with Eu3+ at the high-affinity Ca2+ sites and Cr3+-ATP at the ATP-binding/hydrolytic site. Our results show that ATP-binding/hydrolytic site is separated by about 2.2 nm from each high-affinity Ca2+ site. PMID- 2974805 TI - Restoring the function of a proximally stenosed or occluded sequential aortocoronary venous graft by PTCA of a bypassed native vessel: the 'back-door technique'. AB - If a sequential saphenous vein graft occludes proximally but its distal part remains open, it may act as a large intercoronary collateral. In such situations percutaneous transluminal coronary angioplasty (PTCA) of a stenosis in one of the recipient native vessels may restore the function of the graft. In 250 of our patients who underwent PTCA after previous coronary artery bypass surgery, this technique could be used 20 times in 18 patients. PTCA was performed 3-101 months (mean 24.7 months) after bypass surgery. Complete revascularization could be achieved in all but three patients. Two patients had a small myocardial infarction during PTCA. The primary success rate was 95%. Two patients had a clinical and angiographic recurrence and underwent successful redilatation. Fourteen patients have remained symptom free during follow-up, which ranged from 4 to 44 months (mean 19.5 months). We conclude that this 'back-door technique' provides a new approach to symptomatic relief in patients with recurrent symptoms after previous sequential venous bypass grafting. PMID- 2974806 TI - Immune-deficiency in Hodgkin's disease (HD): a study of patients and healthy relatives in families with multiple cases. AB - In this study we evaluated some immunological features in both patients and healthy relatives within five families with multiple cases of Hodgkin's disease (HD). Such familial groups (at 'high risk' of HD), represent, in our opinion, a suitable opportunity to investigate the role of immune-deficiency in HD. The results obtained in the patient group confirm the well known persistent immune derangement in long term HD survivors. Regarding the group of relatives, we found a pattern similar to that of the patients. In particular, a decrease in the T helper lymphocyte subset and a lower response to Con-A mitogen were detected, which were statistically significant. These findings, confirmed in other studies of multiple case families, could support the hypothesis of a preexisting immune deficiency in HD. This in turn would greatly contribute towards a better understanding of the role of immune-deficiency in the etiopathogenesis of the disease. PMID- 2974807 TI - Young children with orthopedic handicaps: self-knowledge about their disability. PMID- 2974808 TI - Shear strength of orthodontic bonding agents. PMID- 2974809 TI - [Effect of heparin and hypobaric hypoxia on the blood electrolyte composition, ATPase activity and the membrane charge of erythrocytes]. AB - The effects of hypoxia (an "altitude" of 6000 m) and heparin on contents of ions of sodium, potassium, calcium, and magnesium in the plasma and erythrocytes, on ATPase activity and erythrocyte membrane charge in albino rats were studied. Heparin was shown to decrease levels of ions of sodium, potassium, in the plasma and erythrocytes, to increase the negative charge of erythrocytes and membrane permeability for potassium ions. Changes in concentrations of sodium, potassium, calcium and magnesium ions after keeping the rats in a barochamber are more marked on the 2nd day than on the 4th day. Na+, K+-ATPase activity of erythrocyte ghosts first decreased, but on the 4th day increased. Heparin action tended to compensate for changes in contents of the studied ions, ATPase activity and erythrocyte membrane changes as well as changes in gradients of potassium, calcium and magnesium ions concentrations in the erythrocyte-plasma system caused by hypobaric hypoxia. PMID- 2974810 TI - [Determination of irritant dose 50 and irritant time 50 in various study conditions]. PMID- 2974811 TI - Mixed lymphocyte reactions (MLR) in rainbow trout (Salmo gairdneri) sibling. AB - Suitable conditions for 2 way MLR were determined in random combinations of commercially cultured trout and were then applied to full siblings in two families reared for experimental purposes. The primary aim of the investigation was to determine whether the immunogenetic predictions from mammalian and avian experiments would apply, i.e. (i) that all full sibs, short of rare intra-MHC recombinants, can be assigned to a maximum of four different MHC genotypes, and (ii) that individuals of the same genotype are mutually histocompatible as tested in MLR. If both predictions apply, five or more siblings tested in all pairwise combinations should invariably display compatible pairs, so that a maximum of four groups of mutually incompatible individuals should emerge. The findings did not fit this model, and two alternative immunogenetic models are discussed. PMID- 2974812 TI - Partial purification of the D2 dopamine receptor from bovine caudate nucleus. PMID- 2974813 TI - Molecular analysis of two general markers of neurons and the diffuse neuroendocrine system: neuron-specific enolase and PGP9.5. PMID- 2974814 TI - Calcium homoeostasis in excitable cells. PMID- 2974816 TI - Managing cooperative nonambulatory patients: transfers to the dental chair. PMID- 2974817 TI - Dental management of the geriatric alcoholic patient. PMID- 2974815 TI - Adrenal precursor C19 steroids are potent stimulators of growth of androgen sensitive mouse mammary carcinoma Shionogi cells in vitro. AB - Since there is convincing evidence for a role of adrenal steroids as precursors of active sex steroids in peripheral tissues, especially prostate cancer, we have studied the effect of the four main adrenal steroids, namely dehydroepiandrosterone sulfate (DHEA-S), DHEA, 5-androstene-3 beta,17 beta-diol (delta 5-diol) and 4-androstene-3,17-dione (delta 4-dione) on the growth of an androgen-sensitive clone (SEM-1) of the mouse mammary carcinoma Shionogi. From a control doubling time of 6.69 +/- 0.03 days, 0.1 microM DHT, 1.0 microM delta 4 dione, 10 microM delta 5-diol, 10 microM DHEA-S and 10 microM DHEA decreased generation time to 1.60 +/- 0.01, 1.69 +/- 0.01, 1.95 +/- 0.01, 4.37 +/- 0.02 and 5.66 +/- 0.03 days, respectively (P less than 0.01 vs. control). The same compounds exerted their stimulatory effects on cell growth at the following ED50 values: 0.06 nM, 16 nM, 90 nM, 150 nM and 16 microM for DHT, delta 4-dione, DHEA, delta 5-diol and DHEA-S, respectively. The stimulatory effect of all compounds was inhibited in a competitive manner by the pure antiandrogen hydroxyflutamide. Further evidence for an action of the adrenal steroids through the androgen receptor is indicated by competition of [3H]testosterone uptake in the tumor cells at the following IC50 values: 0.21 nM, 0.63 nM, 50 nM, 75 nM and 680 nM for DHT, testosterone, delta 4-dione, delta 5-diol and DHEA, respectively. The present data show that the four main adrenal steroids present in the serum of adult men can exert potent stimulatory effects on the growth of an androgen sensitive cancer cell line through an androgen receptor-mediated mechanism. PMID- 2974818 TI - Intake of energy and nutrients before and after dental treatment in geriatric long-stay patients. PMID- 2974819 TI - Effectiveness of tooth- and denture-brushing in the disabled elderly in municipal old people's homes in Finland. PMID- 2974820 TI - Prediction of need and acceptance of dental services for institutionalized patients. PMID- 2974821 TI - [Follow-up of patients with chloracne in the Seveso area]. AB - Three follow-up surveys from 1976 to 1985 were carried out on 193 subjects who developed chloracne following the Seveso accident (1976). A comparison group, age and sex matched, was selected randomly from the municipal list of Varedo (in the same health district but out of dioxin polluted zone). At each follow-up a questionnaire was administered and biochemical tests, skin examination and electrophysiologic measurements were performed. Biochemical indicators of hepatic function and nerve conduction studies did not show either significant differences between groups or temporal trends. Chloracne was shown to be clinically reversible: all the chloracne cases (except for one subject) clinically recovered by 1983. The discrepancy between our results and the previous experiences can be related to: different type of exposure, target population and interval since exposure. PMID- 2974822 TI - [Occupational contact dermatitis: analysis of clinical case reports, 1972-1985]. AB - The hospital records of 105 subjects with professional dermatitis, admitted at the Institute of Occupational Health of Verona between 1972 and 1985, were reviewed. Work sectors, chemicals, sites of dermatitis, latency and age of onset were evaluated. Contact allergic dermatitis were 80% while irritative ones were 10%. Metal-, construction- and tannery workers were more frequently involved. The most common causes were salts of heavy metals. The authors discuss results and remark the need of standardization in diagnostic procedures. PMID- 2974824 TI - [Conservative organ-preserving measures in precancerous stages of endometrial cancer]. PMID- 2974823 TI - Comparison of the orotidine 5'-monophosphate decarboxylase sequences of eight species. AB - Predicted amino acid sequences of the enzyme orotidine 5'-phosphate decarboxylase (EC 4.1.1.23) from eight different organisms are compared. The comparisons are made on the basis of primary structural differences, primary amino acid sequence, hydropathy profiles, and secondary structure predictions. The organisms compared are Mus musculus, Aspergillus nidulans, Neurospora crassa, Kluyveromyces lactis, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Escherichia coli, and Salmonella typhimurium. PMID- 2974825 TI - Postmortem changes in the level of calcium pumping adenosine triphosphatase in rat heart sarcoplasmic reticulum. AB - The activity of calcium pumping adenosine triphosphatase (Ca2+-ATPase) in cardiac sarcoplasmic reticulum plays a pivotal role in myocardiac contraction-relaxation. The Ca2+-ATPase activity is controlled by phosphorylation and dephosphorylation of a sarcoplasmic reticulum protein "phospholamban" in response to neurotransmitters and drugs. To clarify the role of Ca2+-ATPase in the development of cardiac rigor mortis, we examined the changes of cardiac rigidity and cardiac sarcoplasmic reticulum Ca2+-ATPase activity up to 5 h after the decapitation of rats. Fifteen minutes after decapitation, the rats showed a cardiac rigidity on left ventricles. After 30 min, rigidity was obvious over the whole heart. After 1 h, the rigidity reached a high degree which was maintained for the rest of the observation period. On the other hand, the Ca2+-ATPase activity controlled by phosphorylation and dephosphorylation of phospholamban did not change for the whole observation period (5 h). Another Ca2+-ATPase activity representing the total amount of Ca2+-ATPase in sarcoplasmic reticulum gradually decreased. The data suggest that no significant phosphorylation or dephosphorylation of phospholamban occurs for a short time, at least for 5 h, after death and that the Ca2+-ATPase tends to relax the myocardium against the development of cardiac rigor mortis. PMID- 2974826 TI - Bloodstain characterization in the EAP, Hp, Hb, AK and Glo I typing systems using minigels and the PhastSystem. AB - Application of minigels and the PhastSystem to obtain phenotyping results from bloodstains in the EAP, Hp, AK, and Glo I typing systems was investigated. Nonequilibrium isoelectric focusing with 4-6.5 PhastGel produced readily interpretable phenotypes in the EAP typing system. Both 4-6.5 and 5-8 PhastGel produced AK typing system phenotypes using nonequilibrium isoelectric focusing conditions. The 8-25% PAG PhastGel developed by two staining techniques allowed discrimination of phenotypes for the Hp typing system. Phenotypes from the Glo I typing system were also obtained with this gel type. Variant haemoglobins could be detected on pH 5-8 PhastGel using isoelectric focusing conditions. Much potential for standardized, rapid phenotyping of bloodstains was found to exist utilizing the PhastSystem. PMID- 2974828 TI - Clinical application of percutaneous transluminal angioscopy. Comparison of findings in percutaneous transluminal angioplasty, thrombolysis, thrombus extraction and stent-application. AB - Since the introduction of ultrathin angioendoscopes into clinical application, it has been possible to subject the findings of conventional or digital angiography to objective verification. In dogs and in patients, this new angioscopic method for visualization of arteries without surgical intervention has been carried out. The approach is transfemoral. Endoscopy is combined with angiography and, when indicated, percutaneous transluminal angioplasty (PTA), thrombolysis, thrombus extraction and insertion of a new type of stent. An ultrathin endoscope with an outer diameter of 1.6 mm and a working channel of 0.35 mm is used. Guidewires, contrast media and drugs for local thrombolysis can be employed through this channel. The endoscope is placed in the region of interest under fluoroscopy and direct visualization is obtained by using 0.9% NaCl for decreasing blood flow. Image recording is achieved by video or high-speed camera. With this method, the intraluminal events of dilatation, recanalization, local lysis, stent application and thrombus extraction can be visualized. PMID- 2974827 TI - [Lasers for vascular stenoses? High theoretical expectations--problems in practice: gas bubbles, hemorrhage and catheters]. PMID- 2974829 TI - Activation of phospholipase A2 of human spermatozoa by proteases. AB - The effect of various proteases (kallikrein, plasmin, and trypsin) on sperm phospholipase A2 activity (PA2: EC 3.1.1.4) has been studied. The addition of trypsin to spermatozoa, isolated and washed in the presence of the protease inhibitor benzamidine, increased PA2 activity optimally with trypsin concentrations of 1.0-1.5 units/assay. In kinetic studies, all of the above proteases stimulated the deacylation of phosphatidylcholine (PC); in fresh spermatozoa, trypsin showed a higher activation potential than kallikrein or plasmin. In the presence of benzamidine, the activity remained at basal levels. Endogenous protease activity due to acrosin (control) resulted in an increase in PC deacylation compared to the basal level. The maximum activation time of PA2 activity by proteases was 30 min. Natural protease inhibitors (soybean trypsin inhibitor and aprotinin) kept the PA2 activity at basal levels and a by-product of kallikrein, bradykinin, did not significantly affect the control level. Protein extracts of fresh spermatozoa exhibited the same pattern of PA2 activation upon the addition of proteases, thus indicating that the increase in PA2 activity was not merely due to the release of the enzyme from the acrosome. All of these findings suggest the presence of a precursor form of phospholipase A2 that can be activated by endogenous proteases (acrosin) as well by exogenous proteases present in seminal plasma and in follicular fluid (plasmin, kallikrein). Thus, this interrelationship of proteases and prophospholipase A2 could activate a dormant fusogenic system: the resulting effect would lead to membrane fusion by lysolipids, key components in the acrosome reaction. PMID- 2974830 TI - [Streptozocin-sensitized lymphocytes suppress the glucose-stimulated insulin secretion of the islands of Langerhans]. AB - Isolated islets of Langerhans (from C57Bl/6J mice) were either incubated with culture medium only or medium substituted by 1 mM streptozocin (SZ) and cultivated with lymphocytes (Ly) for seven days. Ly were isolated from the same mouse strain sensitized against SZ by 5 x 40 mg SZ per kg and day intraperitoneally or 1 mg SZ subcutaneously in vivo. Glucose-stimulated insulin secretion rate was used as an indicator for the metabolic condition of islets. SZ pretreated islets showed significantly decreased insulin secretion in the presence of Ly. The secretion capacity of untreated islets was not impaired. PMID- 2974831 TI - Macrophage function in chronic experimental alcoholism. I. Modulation of surface receptors and phagocytosis. AB - In the present study, we have assessed peritoneal, alveolar and splenic macrophages for expression of Fc and C3b surface receptors and their ability to function in immunophagocytosis. We have also measured their oxidative burst response by the nitroblue tetrazolium dye (NBT) reduction method. Our studies revealed that macrophages harvested from chronic alcoholic rats expressed surface C3b and Fc receptors, with significantly higher surface density than macrophages of litter-mate controls (matched for sex and nutritional calories). However, the ability of macrophages from alcoholic rats to phagocytize through C3b and Fc receptors was significantly impaired. In addition, the ability of peritoneal macrophages from alcoholic animals to ingest non-opsonized Candida albicans and to reduce NBT dye was markedly compromised. Abnormalities of macrophage function may, at least in part, account for an increased susceptibility of alcoholic patients to infection. PMID- 2974832 TI - Functional studies on macrophage populations in the airways and the lung wall of SPF mice in the steady-state and during respiratory virus infection. AB - Collagenase digestion of slices of lavaged and perfused murine lung from SPF animals yielded, on average, 104 x 10(6) mononuclear cells per gram tissue, including approximately 14% F4/80+ macrophages and 35% lymphocytes. The lung tissue-associated (digest) macrophage (LDM) population was 10-fold higher in number than the alveolar macrophage (AM) population recoverable by lavage. Comparative functional analysis of these populations was performed, employing assays for immune receptors (Fc and C3), complement-induced spreading, endogenous peroxidase, IL-1 secretion and tumour cytolysis; resident and activated peritoneal macrophages and blood monocytes were examined in parallel. The resident LDM population exhibited an activation status intermediate between blood monocytes and the relatively more activated AM from the airways. Murine lung macrophages were also examined during an acute influenza infection and LDM and AM recoveries increased significantly. The AM population exhibited activation during an acute infection. In contrast, LDM remained quiescent, despite the influx of a large number of T lymphocytes into the lung wall. These results suggest that LDM may be intrinsically resistant to the signals generated during T-cell-mediated eradication of influenza, or else local tissue factors modulate T-cell-derived activation signals. PMID- 2974834 TI - [Acne: epidemiology, motivation and request for treatment]. PMID- 2974833 TI - An immunohistological study of CD4+ lymphocyte subsets within inflammatory lesions with special reference to rheumatoid arthritis and inflammatory bowel disease. AB - The monoclonal antibodies (mAbs) WR16, UCHL1 and WR19 identify subsets of CD4+ lymphocytes that have been functionally characterized as suppressor inducer cells or helper inducer cells. These were applied as components of a panel of lymphocyte-specific mAbs for the phenotypic analysis of lymphocyte populations within biopsies taken from rheumatoid synovial membrane and normal and inflamed gut. The phenotype of peripheral blood lymphocytes from patients with rheumatoid arthritis were also compared to normal controls. The rheumatoid synovium was characterized immunohistologically by a lymphocytic infiltrate composed predominantly of CD4+ lymphocytes and a CD4:CD8 ratio of 2.4. The CD4+ population was composed of UCHL1+ cells to the exclusion of WR16+ cells. This finding was confirmed by double immunofluorescence staining using directly conjugated Leu-3a and WR16. The UCHL1+/WR16-/CD4+ phenotype was maintained in the synovial biopsies regardless of whether the patient had commenced treatment with disease modifying drugs. The absence of WR16+ cells within the rheumatoid synovium was shown to be a localized phenomenon as there was a slight elevation of circulating WR16+ lymphocytes in the peripheral blood of rheumatoids whilst the levels of UCHL1+ and WR19+ lymphocytes remained unchanged. As no appropriate normal control tissue is available for comparison to the rheumatoid synovium we also examined the lymphocytes present within Crohn's disease-involved bowel biopsies and compared them to normal gut tissue lymphocytes using WR16 and UCHL1 mAbs. The CD3+ lymphocytes present within normal tissue comprised a mixture of WR16+ and UCHL1+ cells. In contrast the CD3+ lymphocytes within Crohn's involved tissue were exclusively UCHL1+ as previously observed in the rheumatoid synovium. These data indicate that the CD4+ lymphocyte infiltrate present within inflammatory lesions of presumed distinct aetiology exhibit a localized selective loss of cells with the CD45R+/CD4+ suppressor inducer phenotype. This may be a consequence of the selective extravasation of CD4+ helper induced cells or more likely, in view of the previously documented loss of the p220 molecule identified by CD45R mAbs upon T-cell activation, the result of CD4+ T-cell activation at sites of inflammation. PMID- 2974835 TI - [Papillon-Lefevre syndrome: spontaneous and therapeutically induced course in 2 clinical cases]. PMID- 2974836 TI - The limited impact of percutaneous coronary artery angioplasty on bypass surgery. AB - Although percutaneous transluminal coronary angioplasty has been used successfully now for 10 years, coronary bypass surgery volumes have not diminished greatly. This paper examines the anatomic limitations of percutaneous transluminal coronary angioplasty. Using complete revascularization criteria, angioplasty could be used in only 38% of patients undergoing coronary bypass surgery at the Cleveland Clinic in 1981. The principal anatomic reason for rejecting patients for percutaneous transluminal coronary angioplasty was chronic total occlusion in a vessel suitable for bypass surgery. Angioplasty could be used more frequently in one- and two-vessel bypass patients than in patients receiving more grafts. Percutaneous transluminal coronary angioplasty is currently limited more by its inability to more completely revascularize advanced coronary artery disease than it is from the technical aspects of crossing or dilating stenoses. PMID- 2974837 TI - Pharmaceutical effect of contraceptive pills on the skin. AB - Described are the various effects of birth control methods on the skin-mainly those of the contraceptive pill. The equilibrium of healthy skins and mucosa might be affected by these chemicals or pharmaceutical agents, causing different manifestations. The skin and mucosa should be looked upon as integral parts of the human body and its functions even where localized symptoms arise. PMID- 2974838 TI - The need for a better definition of ADL. AB - In rehabilitation information about the client is an important tool. The main concepts available for this purpose are the negative triad impairment--disability -handicap and the positive term ADL (activities of daily living). Retrospective data from an instruction group for parents of disabled children indicate that the negative triad does not appeal to them, while the positive ADL proves too general a term. Other findings from adult amputees suggest that the negative approach may even hamper efforts to overcome their handicap. So, tripartition of the positive ADL seems necessary. A threefold positive approach indeed proves useful in dealing with clients, but in such cases standard expressions can not be used. It is therefore argued that abbreviations as technical terms in their own right should be introduced for international professional usage, namely: ODL (mere functions, required for ADL performance), ADL proper (the well-known but never defined activities of daily living) and IDL (usual pursuits concretely described, thus remaining both value-scalable and ADL-analysable). More elaborate tentative definitions are provided. PMID- 2974839 TI - Age and pattern of intellectual decline among Down syndrome and other mentally retarded adults. AB - A comparison was made of Down Syndrome and other mentally retarded young adults in community work and residential settings for differences in cognitive profile across age groups using the Wechsler Intelligence Scale for Children. Evidence is of a significant erosion of test scores, especially performance scale scores for the non-DS mentally retarded subjects from the third to fourth decades of life. The Down Syndrome cohort remained relatively stable for cognitive profile over time with the notable exception of the Block Design subtest which declined significantly over the two decades; and in contrast with the control sample. Further investigation is suggested in the event that the Block Design subtest has potential as a diagnostic 'marker' for the early identification of DS individuals likely to be most susceptible to premature Alzheimer's-like senile dementia. PMID- 2974840 TI - Radiofrequency balloon angioplasty. Rationale and proof of principle. AB - Post-angioplasty restenosis (PARS) in atherosclerotic lesions of medium and small arteries occurs in about one-third of cases in the first year following percutaneous transluminal angioplasty (PTA) (early PARS). PARS includes acute spasm, dissection with reclosure, elastic recoil, fibrocellular proliferative response, and progressive atheromatous disease. Fibrocellular proliferation (possibly initiated by platelet derived growth factor) is felt to be culpable in many cases of early PARS (months). Pharmacologic regimens, stents, and thermal welding of the intimal-medial cracks of PTA are among the interventions being developed to deal with PARS. Radiofrequency (RF) current as a source of thermal energy may be useful in combination with balloon angioplasty to reduce PARS. Ideally, this combination would (1) weld intimal-medial cracks of PTA; (2) mold plaque and normal vessel to increase lumen diameters without creating intimal medial cracks; and (3) destroy medial smooth muscle cells and multipotential cells (cellular substrate of PARS). Canine in vivo studies have established the feasibility of RF-mediated vascular tissue welding. Human aortic specimens (N = 28) were manually dissected into intima-media and media-adventitia layers. Bipolar RF energy (650 KHz, total 300 J) and mechanical pressure (1 atm) (experimental group, N = 24) or mechanical pressure alone (control group, N = 4) were applied to the reapposed specimen layers in a special chamber. The chamber was modified with a bipolar electrode designed to reproduce that planned for an RF balloon angioplasty catheter. Welding was demonstrated in normal and atherosclerotic treated specimens (23/24 or 96%) but not controls (0/4).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974841 TI - Sensitivity and specificity of chest roentgenogram criteria for right ventricular hypertrophy. PMID- 2974843 TI - [Back problems during pregnancy. 2 physiotherapists' viewpoints of the problem]. PMID- 2974842 TI - Measurement of colloidal iron binding at low pH in cartilage using the proton microprobe. AB - Quantitative micro-PIXE analysis was performed on mouse embryo epiphyseal cartilage and on the rib cartilage of mature animals after incubation of sections with colloidal iron at pH 1.8. The iron content as well as that of sulphur and phosphorus and Fe/S, Fe/P ratios were determined. It was found that colloidal iron content was higher in the cartilage than in other tissues. The cartilage also displayed the highest content of sulphur. The Fe/S ratio was however not constant, being highest in the degeneration zone close to the mineralization front, where the binding of iron was strongest while the amount of sulphur decreased. This indicates that factors other than number of sulphate groups influence the binding of positively charged molecules to glycosaminoglycans. This is confirmed by differences in the results obtained for embryonic and mature rib cartilage. PMID- 2974844 TI - Atrial natriuretic peptide in acute mountain sickness. AB - To test the hypothesis that elevated atrial natriuretic peptide (ANP) may be involved in altered fluid homeostasis at high altitude, we examined 25 mountaineers at an altitude of 550 m and 6, 18, and 42 h after arrival at an altitude of 4,559 m, which was climbed in 24 h starting from 3,220 m. In 14 subjects, symptoms of acute mountain sickness (AMS) were absent or mild (group A), whereas 11 subjects had severe AMS (group B). Fluid intake was similar in both groups. In group B, urine flow decreased from 61 +/- 8 (base line) to 36 +/- 3 (SE) ml/h (maximal decrease) (P less than 0.05) and sodium excretion from 7.9 +/- 0.9 to 4.6 +/- 0.7) mmol.l-1.h-1 (P less than 0.05); ANP increased from 31 +/ 4 to 87 +/- 26 pmol/l (P less than 0.001), plasma aldosterone from 191 +/- 27 to 283 +/- 55 pmol/l (P less than 0.01 compared with group A), and antidiuretic hormone (ADH) from 1.0 +/- 0.1 to 2.9 +/- 1.2 pmol/l (P = 0.08 compared with group A). These variables did not change significantly in group A, with the exception of a decrease in plasma aldosterone from 189 +/- 19 to 111 +/- 17 pmol/l (P less than 0.01). There were no measurable effects of elevated ANP on natriuresis, cortisol, or blood pressure. The reduced diuresis in AMS may be explained by increased plasma aldosterone and ADH overriding the expected renal action of ANP. The significance of elevated ANP in AMS remains to be established.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974845 TI - Involvement of ANF in the acute antidiuresis during PEEP ventilation. AB - To investigate the potential role of natriuretic factor (ANF) on changes on renal excretory function in response to increased intrathoracic pressure, seven patients were studied during three successive 60-min periods of 1) mechanical ventilation (MV) and zero end-expiratory pressure (ZEEP), 2) MV with 12 cmH2O positive end-expiratory pressure (PEEP), and 3) MV with the same level of PEEP while lower-body positive pressure (LBPP) was applied to restore venous return and increase central blood volume without fluid loading. Hemodynamics, renal excretory function parameters, and plasma immunoreactive atrial natriuretic factor (irANF) levels were recorded at the end of each period. Compared with ZEEP, PEEP induced a significant reduction of diuresis (from 134 +/- 17 to 59 +/- 13 ml/h, P less than 0.01) and natriuresis (from 8.37 +/- 3.5 to 3.83 +/- 2 mmol/h, P less than 0.01), whereas plasma irANF fell from 520 +/- 292 to 155 +/- 40 pg/ml (P less than 0.01) and transmural right atrial pressure decreased from 3.9 +/- 0.5 to 2.4 +/- 0.3 mmHg (P less than 0.01). Opposite changes were observed during application of LBPP, which restored diuresis and plasma irANF to near control ZEEP values, despite continuation of PEEP. Changes in renal excretory function parameters thus paralleled changes in right atrial pressure and plasma irANF. We suggest that changes in plasma irANF in response to hemodynamic variations induced by changes in intrathoracic pressure may contribute to alterations of renal excretory function during PEEP. PMID- 2974846 TI - Atrial natriuretic factor attenuates the pulmonary pressor response to hypoxia. AB - The influence of endogenous and exogenous atrial natriuretic factor (ANF) on pulmonary hemodynamics was investigated in anesthetized pigs during both normoxia and hypoxia. Continuous hypoxic ventilation with 11% O2 was associated with a uniform but transient increase of plasma immunoreactive (ir) ANF that peaked at 15 min. Plasma irANF was inversely related to pulmonary arterial pressure (Ppa; r = -0.66, P less than 0.01) and pulmonary vascular resistance (PVR; r = -0.56, P less than 0.05) at 30 min of hypoxia in 14 animals; no such relationship was found during normoxia. ANF infusion after 60 min of hypoxia in seven pigs reduced the 156 +/- 20% increase in PVR to 124 +/- 18% (P less than 0.01) at 0.01 microgram.kg-1.min-1 and to 101 +/- 15% (P less than 0.001) at 0.05 microgram.kg 1.min-1. Cardiac output (CO) and systemic arterial pressure (Psa) remained unchanged, whereas mean Ppa decreased from 25.5 +/- 1.5 to 20.5 +/- 15 mmHg (P less than 0.001) and plasma irANF increased two- to nine-fold. ANF infused at 0.1 microgram.kg-1.min-1 (resulting in a 50-fold plasma irANF increase) decreased Psa (-14%) and reduced CO (-10%); systemic vascular resistance (SVR) was not changed, nor was a further decrease in PVR induced. No change in PVR or SVR occurred in normoxic animals at any ANF infusion rate. These results suggest that ANF may act as an endogenous pulmonary vasodilator that could modulate the pulmonary pressor response to hypoxia. PMID- 2974847 TI - Influence of spaceflight on rat skeletal muscle. AB - The size, succinate dehydrogenase (SDH) and alpha-glycerolphosphate dehydrogenase (GPD) activities, and alkaline myofibrillar adenosinetriphosphatase (ATPase) staining properties were determined from quantitative histochemical analyses of single fibers from five hindlimb muscles of six male rats exposed to a 7-day National Aeronautics and Space Administration spaceflight mission (SL-3). These same properties were determined in a group of ground-based control rats housed under simulated environmental conditions. The wet weight of each of the flight muscles was significantly reduced relative to control. However, the loss of mass varied from 36% in the soleus to 15% in the extensor digitorum longus. The cross sectional areas of fibers in the flight muscles also were reduced, except for the dark ATPase fibers in the medial gastrocnemius. The greatest relative fiber atrophy occurred in the muscles with the highest proportion of light ATPase fibers. An increase in the percentage of dark ATPase fibers also was observed in flight muscles with a predominance of light ATPase fibers. Also, there was an increase in the biochemically determined myofibrillar ATPase activity of tissue sections of the flight soleus. No changes in histochemical or biochemical measures of ATPase activity were observed in the flight extensor digitorum longus. In general, the SDH activity of flight muscles was maintained, whereas GPD activity either was maintained or increased. Based on a metabolic profile of ATPase, SDH, and GPD, there was an increase in the proportion of fast oxidative glycolytic fibers in some muscles. PMID- 2974848 TI - Spectrum analysis of turbulence in the canine ascending aorta measured with a hot film anemometer. AB - We measured turbulence velocity in the canine ascending aorta using a hot-film anemometer. Blood flow velocity was measured at various points across the ascending aorta approximately 1.5-2 times the diameter downstream from the aortic valve. The turbulence spectrum was calculated and its characteristics were examined in connection with the mean Reynolds number and/or measuring positions. In the higher wave number range the values of the turbulence spectra were higher at larger mean Reynolds number. In the higher wave number range, the values of the turbulence spectra were higher at points closer to the centerline of the aorta, when the mean Reynolds number was relatively large. The patterns of the turbulence spectra at various points outside the boundary layer on the aortic wall were similar. PMID- 2974849 TI - Outcome in patients with failed percutaneous transluminal angioplasty for peripheral vascular disease. AB - The outcome in 299 patients having 321 percutaneous transluminal angioplasty (PTA) procedures for peripheral vascular disease was analysed. Technical failure occurred in 21 patients (7%) but in none was the limb ischemia made worse by the failed PTA attempt; nine of these (3%) had been considered unsuitable for arterial reconstruction and proceeded to primary amputation, while 12 (4%) did not have subsequent management compromised by the failed PTA attempt. Complications occurred in seven patients (2.3%); four of these (1.3%) had worsening ischemia but were able to be satisfactorily managed by surgical intervention. There were 71 patients (23.7%) who had an initially successful PTA procedure which subsequently failed; 20 of these (6.7%) had been considered unsuitable for arterial reconstruction and proceeded to amputation, while five patients suitable for arterial reconstruction (1.7%) came to amputation, four following failed bypass surgery and one following multiple trauma from a motor vehicle accident. The remaining 46 patients (15.3%) did not have subsequent management compromised by the late failure of PTA. Early and late failure of PTA in patients presenting with peripheral vascular disease does not compromise subsequent management. PMID- 2974850 TI - Mixed lymphocyte reactions do not predict severity of graft versus host disease (GVHD) in HLA-DR compatible, sibling bone marrow transplants. AB - Mixed lymphocyte reactions (MLRs) were measured in 25 HLA-A, B and DR compatible sibling bone marrow transplants. Only four of 25 MLRs were positive and in these the low reactivity was of doubtful clinical importance. There was no correlation between MLR and the subsequent development or severity of graft versus host disease (GVHD). A survey of bone marrow transplant units in the United Kingdom showed that most centres perform HLA-DR typing as well as an assessment of the MLR. Factors other than histocompatibility are important in the pathogenesis of GVHD and the data from this study suggest that conventional MLRs can be omitted in HLA-A, B and DR compatible sibling bone marrow transplants. PMID- 2974851 TI - Effects of cyproterone acetate, LHRH agonist and ovarian surgery in McCune Albright syndrome with precocious puberty and galactorrhea. AB - We have studied the endocrinological pattern in a girl with McCune-Albright syndrome. The young patient showed: normal prepubertal serum levels of gonadotropins, fluctuating estrogen concentrations, which sometimes were similar to the levels in adult women of fertile age, hyperprolactinemia with galactorrhea, ovarian cysts. The effects of treatment with antiandrogen drug, cyproterone acetate, and of a LHRH agonist, buserelin (less than D-Ser[TBU(8)6 des-gly NH2.10 greater than LHRH ethylamide), were studied. Cyproterone acetate with or without buserelin did not fully suppress estradiol concentrations. On the other hand, surgical resection of these cysts resulted in both clinical and endocrinological remission. It is likely that in this case of McCune-Albright syndrome precocious puberty was a result of ovarian estrogen secretion, while pubertal activation of the hypothalamus-pituitary axis was absent. Hyperprolactinemia, which appeared after the beginning of the combined therapy with buserelin and cyproterone acetate, was probably due to the elevated estrogen levels. PMID- 2974852 TI - The treatment of a hyperandrogenic and virilizing state in an elderly female with a synthetic LHRH agonist. AB - A case of hyperandrogenism and virilization is described in an elderly female. She had elevated testosterone levels, but normal DHEAS and 24-h urinary 17 oxosteroid excretion, suggesting an ovarian tumor. Stimulation and suppression tests, and radioisotopic and radiological scans proved unhelpful in the diagnosis although hyperthecosis of the ovary was later suggested by ultrasound. Testosterone and gonadotropin levels fell during therapy with cyproterone acetate and subsequently ethinyl estradiol. Because of side effects encountered on these drugs, she was treated with a synthetic, slow-release preparation of an LHRH agonist, D-TRP-6-LHRH (Decapeptyl), with symptomatic and biochemical improvement. Long term LHRH agonists might be a valuable treatment for hyperandrogenic states in elderly patients. PMID- 2974853 TI - [Medicolegal risks of celioscopy. Analysis of 32 cases of complications]. AB - An analysis of 200 case records of complications or undesired incidents that happened in the practice of gynaecological surgery and which often gave rise to legal actions either in the criminal or the civil courts, made it possible to extract from these case notes 32 cases where there were medico-legal consequences from laparoscopies that had become complicated. Above all, the relative seriousness of each case had to be considered. More than a third of these cases led to death directly linked to the gynaecological laparoscopy. This furthermore could be responsible for intestinal or vascular complications that had to be treated by intensive resuscitation and by further operative interference carried out as emergencies and by prolonged hospitalisation. After having reported these 32 case histories in detail, the authors analyse the medical consequences of these complications before going on the legal repercussions arising from them. The medico-legal practices that are so important are the ability of the laparoscopist, the standards of his technical equipment and of the anaesthetist, and his taking note of previous operations that have been carried out on the patient. It should not be forgotten, too, that there is a statistically demonstrated risk when laparoscopy sterilization is carried out, because this itself gives rise to claims. Finally, the preventive measures that can be undertaken in order to protect surgeons are outlined. These follow the discovery of an important series of complications which may help them to avoid trouble in their daily practice. PMID- 2974854 TI - [A comparative study of diltiazem blood cardioplegia in 49 patients undergoing aortic valve replacement with left ventricular hypertrophy]. PMID- 2974855 TI - [Accelerated myocardial metabolic and functional recovery with terminal nicorandil-mg cardioplegia in experimental heart transplantation]. PMID- 2974856 TI - [Plasma level of atrial natriuretic polypeptide during and after cardiopulmonary bypass]. PMID- 2974857 TI - [A study of the expression of class II antigen on uterine cervical cancer cells and cell lines]. AB - An immunohistochemical method with monoclonal antibody to DR antigen was used to study class II antigens and HLA-DR antigen in 20 patients with uterine cervical cancer. Eight patients had various amounts of DR antigen. Regional infiltrating lymphocytes were also examined with anti-Leu 1, Leu 2a, Leu 3a, and Leu 10. Among the many infiltrating T cells, Leu 3a-positive cells were relatively predominant surrounding the cancer nests which contained DR antigen. Cervical cancer cell lines OMC-1 and OMC-4 both demonstrated DR antigen. Interferon (IFN)-gamma treatment enhanced the expression of DR antigen in OMC-1 and OMC-4. The DR antigen in OMC-1 and OMC-4 were capable of stimulating allogenic lymphocytes in mixed lymphocyte reaction (MLR), and their stimulatory activity was significantly enhanced by IFN-gamma treatment. These results demonstrated the expression of class II antigen, especially DR, on some cervical cancer cells and cell lines and showed that the DR antigen in uterine cancer cell lines can stimulate MLR. PMID- 2974858 TI - [Comparative study of three antiestrogens in MCF-7 and ZR-75-1 human breast cancer cells]. PMID- 2974860 TI - Regulation of binding of phosphofructokinase to myofibrils in the red and white muscle of the barred sand bass, Paralabrax nebulifer (Serranidae). AB - The binding of phosphofructokinase (PFK) to myofibrils from the white muscle of the fish Paralabrax nebulifer (Girard, 1854) is sensitive to factors known to be allosteric regulators of PFK activity. PFK in Triton-X-100-extracted muscle remains bound to myofibrils at pH 7.0 and is fully solubilized by increasing the pH to 8.0. The curve describing the pH-dependence of PFK binding to myofibrils is similar in its steepness to pH versus activity curves of PFK at low temperature. Nucleotides are also potent modulators, preventing the association of PFK with myofibrils at concentrations between 20 and 60 mumol l-1 of ATP, ADP, MgATP or GTP, listed in order of effectiveness. PFKs in the red and white muscle extracts differ in their pH-dependence of binding to myofibrils, and their kinetic and regulatory properties (response to citrate, pH and fructose-2,6-bisphosphate). Reversible binding of PFK to myofibrils may be important in the control of glycolysis, especially in the highly glycolytic white muscle fibres. PMID- 2974859 TI - Down's syndrome with lingual tonsil hypertrophy producing sleep apnoea. AB - A 12-year-old girl is presented with Down's syndrome and lingual tonsillitis causing sleep apnoea. In order to overcome her sleep apnoea she adopted a functional sleeping posture. The sleep apnoea was successfully treated by a single staged lingual tonsillectomy without preliminary tracheostomy. A return to a normal sleeping posture occurred spontaneously in the early post-operative period. PMID- 2974861 TI - A molecular basis for synexin-driven, calcium-dependent membrane fusion. AB - Membranes of secretory vesicles fuse with each other and with plasma membranes during exocytosis in many different cell types. The probable role of calcium in the process is now widely accepted, and it is possible that at least one cytosolic mediator of calcium action is synexin. Synexin is a 47,000 Mr calcium binding protein, initially discovered in the bovine adrenal medulla, which binds to granule membranes and to inner aspects of chromaffin cell plasma membranes. Synexin causes chromaffin granules to aggregate, and such aggregates can be caused to fuse in the additional presence of arachidonic acid. Synexin also mediates the direct fusion of liposomes and chromaffin granule ghosts. To understand better the mechanisms of membrane fusion promoted by synexin we have attempted to define the primary sequence of the protein. Our initial efforts were directed towards purification of bovine synexin in sufficient amounts to allow us to sequence tryptic peptides. However, as the project progressed we also directed our attention to human synexin, preparing peptides from this protein as well. From analysis of bovine peptides we learned that the synexin molecule might be closely related to a class of proteins including lipocortin I, calpactin (p36), endonexin II, protein II and calelectrin 67K. Complete analysis of a human synexin cDNA clone revealed strong homology with bovine synexin. The analysis also showed that synexin contained a unique, long, highly hydrophobic N-terminal leader sequence followed by a characteristic four-fold repeat homologous with those found in other members of the synexin gene family. The highly hydrophobic character of synexin seems consistent with information previously obtained that synexin is able to insert directly into the interior of bilayers prepared not only from purified phosphatidylserine but also from biological membranes. The evidence for such insertions is a dramatic increase in the capacitance of the membrane, formed at the tip of a patch pipette, when calcium-activated synexin is applied to the bilayer. Additional evidence is the fact that synexin also forms calcium-selective channels when the protein is applied to the cytosolic aspect of the plasmalemma when that side is also exposed to calcium at sub-millimolar concentrations. Thus, the synexin molecule not only enters the membrane, but also spans it. From these and other data we have developed the concept that the fusion process may involve synexin forming a 'hydrophobic bridge' between two fusing membranes. Lipid movement across this bridge may then be the material basis for final fusion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2974862 TI - The distinction between integral and separable dimensions: evidence for the integrality of pitch and loudness. AB - Six experiments are reported that investigated the reality and generality of dimensional integrality (Garner, 1974a) by evaluating whether the auditory dimensions of pitch and loudness are psychologically privileged and whether they combine in an integral fashion. In Experiment 1 we psychophysically scaled the dimensions to ensure that, within the stimulus range used, the perceived value on each dimension would remain constant in the face of variation on the other dimension. In Experiments 2 through 4 we assessed performance by using the converging operations by which Garner defined integrality and separability. Experiment 2 showed that in speeded classification, pitch and loudness lead to facilitation with redundant variation and interference with orthogonal variation. Experiment 3 showed that unspeeded classifications are guided predominantly by overall similarity. Experiment 4 established that the better-fitting metric by which multidimensional similarity is appreciated is Euclidean rather than city block. These results suggest that the dimensions of pitch and loudness combine in an integral fashion. In Experiments 5 and 6 we investigated whether the dimensions of pitch and loudness have a privileged status by assessing the impact of rotating the dimensional axes on performance in a speeded sorting task. Experiment 5 looked at six alternative dimensional orientations to pitch and loudness. If anything, rotating the dimensional axes increased the amount of interference in filtering. In Experiment 6 we assessed an alternative dimensional description of the stimuli based on the dimensions of volume and brightness. We found greater interference when the stimuli varied along the dimensions of volume and brightness than when they varied along the dimensions of pitch and loudness. The fact that the least interference is observed when the stimuli vary along the dimensions of pitch and loudness suggests that these dimensions are the more psychologically valid ones. These findings indicate that integrality is not a "myth," that is, merely a case of psychophysical mismatch. Instead, dimensions that are psychologically real are sometimes processed in a unitary fashion. PMID- 2974863 TI - Phenomenal characteristics of memories for perceived and imagined autobiographical events. AB - Two studies explored potential bases for reality monitoring (Johnson & Raye, 1981) of naturally occurring autobiographical events. In Study 1, subjects rated phenomenal characteristics of recent and childhood memories. Compared with imagined events, perceived events were given higher ratings on several characteristics, including perceptual information, contextual information, and supporting memories. This was especially true for recent memories. In Study 2, subjects described how they knew autobiographical events had (or had not) happened. For perceived events, subjects were likely to mention perceptual and contextual details of the memory and to refer to other supporting memories. For imagined events, subjects were likely to engage in reasoning based on prior knowledge. The results are consistent with the idea that reality monitoring draws on differences in qualitative characteristics of memories for perceived and imagined events (Johnson & Raye, 1981) and augment findings from more controlled laboratory studies of complex events (Johnson & Suengas, in press; Suengas & Johnson, 1988). PMID- 2974864 TI - Representing statics as forces in equilibrium. AB - Resting objects can be described according to the physical forces operating on them, forces that are balanced in static scenes. We hypothesized that in a related way, the perception of static scenes and objects might involve a representation of underlying dynamics. In our first experiments, subjects were shown a picture of a plant resting upon a table or hanging from a hook, followed by a picture of the plant in the same position without the supporting table or hook. Subjects attempted to remember the position of the plant and were then shown a third display, in which the plant was in the same position or was slightly above or slightly below the original position. We found that subjects made more errors for test displays showing the plant slightly below, as compared with displays showing the plant slightly above, the original position. That is, memory for the position of the previously supported object was distorted in the direction consistent with what would happen if the plant was to lose its source of support in real life. This effect depends on the initial display of support; in Experiment 2 we found no memory asymmetry when the plant was initially displayed without support. We replicated the results of Experiments 1 and 2 with a new stimulus set and modified procedure in Experiment 3. In our fourth study we experimented with a slightly different stable situation: a spring with a box on top of it. We found that subjects misremembered the spring as either more compressed or less compressed as predicted by the implied dynamics of the display sequence. We discuss issues raised by our findings, including the possibility that the conscious experience of concreteness in static scenes stems from the representation of underlying forces. PMID- 2974865 TI - Ambiguity in perception and experimentation. AB - Bruno and Cutting (1988) varied four monocular cues to perceived depth in a factorial design. Subjects judged the distance between test objects. Given main effects in the analysis of variance, the authors concluded that the perceivers integrated the four different sources of information, as opposed to simply selecting a single source. Given no interactions in the analysis of variance, the authors concluded that the integration process was additive rather than multiplicative. The ambiguity inherent in Bruno and Cutting's experiments and analyses is discussed. As presented, their results did not provide evidence for integration of depth cues or evidence for additivity, independence, and parallel processing of the cues. An additional analysis of the distribution of the rating judgments given by their subjects, however, provides some evidence for integration of the cues. The fuzzy logical model of perception (FLMP) is extended to describe perceptual recognition of depth. The model assumes independence of the cues during feature evaluation and a nonadditive integration process in which the least ambiguous cues have the greatest impact on the judgment. The FLMP is contrasted with a model assuming additivity of the cues. Because both models describe the results equally well, it remains for future researchers to provide definitive tests between the models. PMID- 2974866 TI - Additivity, subadditivity, and the use of visual information: a reply to Massaro (1988). AB - Previously we (Bruno & Cutting, 1988) explored the perception of spatial relations among objects laid out in a computer-generated environment. In his commentary on our article, Massaro (1988) raised several issues. The most important is from his reanalysis, which indicated that--because of a subadditive trend in the results--additive and multiplicative strategies fit our data in Experiment 1 about equally well. In reply, we performed a different analysis. Results corroborate subadditivity--and hence multiplicative information combination--in Experiment 1 but provide no evidence for it in Experiments 2 and 3. On the whole, then, the results still support additivity more strongly than any other combination rule and thus support our notion of minimodularity. PMID- 2974867 TI - Visual-haptic perceptual nonequivalence for shape information and its impact upon cross-modal performance. AB - The issues of visual-haptic perceptual equivalence and the impact of nonequivalence upon cross-modal performance were explored. A measure of cross modal nonequivalence was developed from multidimensional scaling models of the perceptual structures of 24 nonrepresentative three-dimensional stimuli. In Experiment 1 the visual and haptic perceptual structures and measures of cross modal nonequivalence were shown to be replicable. Experiments 2 and 3 employed sets of stimuli selected as cross modally similar or dissimilar (based upon the results of Experiment 1) and tested the impact of perceptual nonequivalence upon cross-modal performance with shape information. The experiments used somewhat different tasks and produced converging results. There was poorer cross-modal performance when cross modally dissimilar stimuli were involved than when only cross modally similar stimuli were involved, but there was no such pattern for intramodal performance. The findings are related to the theoretical notions of perceptual equivalence (Gibson, 1966; Marks, 1978) and the theoretical and practical importance of understanding the perceptual properties of stimuli used in cognitive tasks (Garner, 1970; Monahan & Lockhead, 1970). PMID- 2974868 TI - Hidden figures are ever present. AB - Preference judgments about alternative interpretations of unambiguous patterns can be explained in terms of a rivalry between a preferred and a second-best interpretation (cf. Leeuwenberg & Buffart, 1983). We tested whether this second best interpretation corresponds to a suppressed but concurrently present interpretation or whether it merely reflects an alternative view that happens to be preferred less often. Two patterns were present immediately following each other with a very short onset asynchrony: a complete pattern and one out of three possible subpatterns of it, corresponding to the best, the second best, or an odd interpretation of the complete pattern. Subjects indicated which subpattern was presented by choosing among the three subpatterns shown after each trial. The scores, corrected for response-bias effects, indicated a relative facilitation of the second-best interpretation, in agreement with its predicted "hidden" presence. This result is more in line with theories that capitalize on the quality of the finally selected representation than with processing models aimed at reaching one single solution as fast and as economically as possible. PMID- 2974869 TI - The observer-relative velocity field as the basis for effective motion parallax. AB - Earlier studies of motion parallax found unambiguous relative depth perception when random dot patterns were systematically translated in accordance with either motion of the observer's head or motion of the display scope. The need for such relative motion between an observer and a flow field was examined by placing a flow field in a limited area (window) in a large scope and translating the window relative to the observer. Accuracy in judging surface orientation and quantitative depth estimates were determined by the velocity field relative to the observer and were not measurably affected by whether this field was produced with a stationary or a moving window. Accuracy was consistently higher for smaller ratios of maximum to minimum projected velocities, reaching 100% in one experiment with a 1.12:1 ratio. We conclude that fully effective motion parallax does not require relative motion between the observer's head and the contours of a flow field. PMID- 2974870 TI - Visual selection from multielement displays: measuring and modeling effects of exposure duration. AB - In a partial-report experiment, subjects reported the digits from a circular array of digits and letters terminated by a pattern mask. Individual frequency distributions of the number of correctly reported digits were analyzed as functions of number of digits (2, 4, or 6) and number of letters (0, 2, 4, 6, or 8) at nine exposure durations ranging from 10 to 200 ms. The distributions (hundreds of data points per subject) were accurately predicted by a four parameter fixed-capacity independent race model that assumes exponentially distributed processing times, limitations in both processing capacity and storage capacity, and time-invariant selectivity. Estimated from the data, processing capacity C was 45 items/s, selectivity alpha (ratio between the amount of processing capacity devoted to a distractor and the amount devoted to a target) was 0.48, short-term storage capacity K was 3.5 items, and the longest ineffective exposure duration t0 was 18 ms. PMID- 2974871 TI - Lexical ambiguity and the timecourse of attentional allocation in word recognition. AB - In two experiments the allocation of attention during the recognition of ambiguous and unambiguous words was investigated. In Experiment 1, separate groups performed either lexical decision, auditory probe detection, or their combination. In the combined condition probes occurred 90, 180, or 270 ms following the onset of the lexical-decision target. Lexical decisions and probe responses were fastest for ambiguous words, followed by unambiguous words and pseudowords, respectively, which indicated that processing ambiguous words was less attention demanding than unambiguous words or pseudowords. Attention demands decreased across the timecourse of word recognition for all stimulus types. In Experiment 2, one group performed the lexical-decision task alone, whereas another group performed the lexical-decision task during the retention interval of a short-term memory task. The results were consistent with those from Experiment 1 and showed that word recognition is an attention-demanding process and that the demands are inversely related to the number of meanings of the stimulus. These results are discussed with regard to the structure of the mental lexicon (i.e., single vs. multiple lexical entries) and the effect of such a structure on attentional mechanisms. PMID- 2974872 TI - Preparation for grasping an object: a developmental study. AB - The development of visually controlled grasping actions was studied in two experiments. An optoelectronic technique (SELSPOT) was used to monitor the opening and closing of the hand during reaching actions by measuring the change in the distance between thumb and index finger. The purpose of Experiment 1 was to establish an adult criterion for the development. It was shown that adults started closing the hand around the target well before touch and that the timing was dependent on the size of the target. The hand started to close earlier when grasping a small rather than a large target. In addition, the degree of hand opening was also less for a small than for a large target. In Experiment 2 it was shown that infants who were 5-6, 9, and 13 months of age also controlled their grasping actions visually and started closing the hand around the target in anticipation of the encounter rather than as a reaction to the encounter. However, the strategy of the two younger age groups was different from that of adults. They started closing the hand closer to the time of contact with the target than did the 13-month-olds, who were comparable to adults in this respect. The timing was not dependent on the size of the target in any of the infant groups. In all age groups, reaching and grasping were most commonly organized in a continuous way; that is, the hand started to close without any interruption in the approach. The opening of the hand was found to be adjusted to target size in the 9- and 13-month-olds but not in the 5-6 month olds. PMID- 2974873 TI - Relation between velocity and curvature in movement: equivalence and divergence between a power law and a minimum-jerk model. AB - Unconstrained hand movements typically display a decrease in hand speed around highly curved sections of a trajectory. It has been suggested that this relation between tangential velocity and radius of curvature conforms to a one-third power law. We demonstrate that a one-third power law can be explained by models taking account of trajectory costs such as a minimum-jerk model. Data were analyzed from 6 subjects performing elliptical drawing movements of varying eccentricities. Conformity to the one-third power law in the average was obtained but is shown to be artifactual. It is demonstrated that asymmetric velocity profiles may result in consistent departures from a one-third power law but that such differences may be masked by inappropriate analysis procedures. We introduce a modification to the original minimum-jerk model by replacing the assumption of a Newtonian point mass with a visco-elastic body. Simulations with the modified model identify a basis for asymmetry of velocity profiles and thereby predict departures from a one-third law commensurate with the empirical findings. PMID- 2974875 TI - Two forms of persistence in visual information processing. AB - Iconic memory, which was initially regarded as a unitary phenomenon, has since been subdivided into several components. In the present work we examined the joint effects of two such components (visible persistence and the visual analog representation) on performance in a partial report task. The display consisted of 15 alphabetic characters arranged around the perimeter of an imaginary circle on the face of an oscilloscope. The observer named the character singled out by a bar-probe. Two factors were varied: exposure duration of the array (10, 50, 100, 150, 200, 300, 400 or 500 ms) and duration of blank period (interstimulus interval, ISI) between the termination of the array and the onset of the probe (0, 50, 100, 150, or 200 ms). Performance was progressively impaired as both exposure duration and ISI were increased. The results were explained in terms of a probabilistic combinatorial model in which the timecourses of visible persistence and of the visual analog representation are regarded as time-locked to the onset and to the end of stimulation, respectively. The impairing effect of exposure duration was attributed to the relatively high spatial demands of the task that could be met optimally by information in visible persistence (which declines as a function of exposure duration), but less adequately by information in the visual analog representation. A second experiment, employing a task with lesser spatial demands, confirmed this interpretation. PMID- 2974874 TI - Perception of translational heading from optical flow. AB - Radial patterns of optical flow produced by observer translation could be used to perceive the direction of self-movement during locomotion, and a number of formal analyses of such patterns have recently appeared. However, there is comparatively little empirical research on the perception of heading from optical flow, and what data there are indicate surprisingly poor performance, with heading errors on the order of 5 degrees-10 degrees. We examined heading judgments during translation parallel, perpendicular, and at oblique angles to a random-dot plane, varying observer speed and dot density. Using a discrimination task, we found that heading accuracy improved by an order of magnitude, with 75%-correct thresholds of 0.66 degrees in the highest speed and density condition and 1.2 degrees generally. Performance remained high with displays of 63-10 dots, but it dropped significantly with only 2 dots; there was no consistent speed effect and no effect of angle of approach to the surface. The results are inconsistent with theories based on the local focus of outflow, local motion parallax, multiple fixations, differential motion parallax, and the local maximum of divergence. But they are consistent with Gibson's (1950) original global radial outflow hypothesis for perception of heading during translation. PMID- 2974877 TI - Comparison requirements and attention in identical-nonidentical stimulus discriminations. AB - Perceptual matching data show several puzzling effects. Particularly problematic are the disparities between the processing rates for same and different stimuli- the fast-same effect--and between the processing rates for two same-different judgment tasks that are related as mirror images--the task effect. Current models have difficulty accounting simultaneously for both effects. Central to these models is a stimulus comparison process that derives relative judgments of sameness and difference from tests of the congruence of stimulus representations. A contrasting view holds that same-different judgments can be modeled as absolute, rather than relative, judgments. This latter view is shown to be supported by experimental data. Reaction times for judgments of identical letter strings increase with string length at the same rate whether judgments are based on all the information in the strings or just the information in a single pair of component letters. The data show that stimulus comparisons of the sort described by previous models are not involved in these judgments. An attentional model accounts for the data and for the fast-same and task effects as well. PMID- 2974876 TI - Use of partial stimulus information in response processing. AB - We examined the reaction time benefit that is obtained when salient features of the stimulus set and response set correspond. Components of the event-related brain potentials were used to measure the timing of stimulus-related and response related processes in order to determine the locus of this effect. Of particular importance was the development of a new index of selective response preparation, the corrected motor asymmetry (CMA). We found no evidence for the use of preliminary, partial stimulus information in response preparation. These results suggest that the benefit is located primarily in response selection processes and probably reflects a more efficient algorithm for stimulus-response translation. Also, we found trial-to-trial variability in the duration of response selection to be the major determinant of variability in reaction time, whereas the durations of subsequent response-related processes were relatively invariant. Implications of these results for discrete and continuous models of choice reaction performance are discussed. PMID- 2974878 TI - Mental curve tracing with elementary stimuli. AB - It has been proposed that certain spatial relations are determined by an operation, or "visual routine," that can trace along a boundary (Ullman, 1984). This proposal was supported by Jolicoeur, Ullman, and Mackay's (1986) finding that the time required to determine if two Xs are on the same curve increased monotonically with the separation of the Xs along that curve. In the present study the generality of the curve tracing hypothesis was explored across four experiments by using elementary stimuli that eliminated interweaving curves, displaced the fixation point away from the curves and target Xs, and provided a simple alternative to curve tracing--namely, determining whether or not the Xs fell on the same side of the figure. Stimuli consisted of two curves (150 degrees arcs) and two Xs, and each stimulus was presented for 150 ms. In Experiments 1 and 2, subjects were instructed to decide as quickly as possible if the two Xs fell on the same curve or on different curves. Even for these elementary stimuli, mean reaction time (RT) for same trials increased monotonically with the distance separating the Xs. Mean RT for different trials, however, decreased with the distance separating the Xs. In Experiments 3 and 4 alternatives to curve tracing were tested. For same trials the evidence strongly favored curve tracing. However, different trials were apparently solved on the basis of judgmental processes presumably operating in parallel with curve tracing. Curve tracing rates fluctuated across experiments and seemed to be partially governed by the width of the "pathway" provided for the trace. PMID- 2974879 TI - In vivo labelling of rat brain dopamine D-2 receptors. Stereoselective blockade by the D-2 antagonist raclopride and its enantiomer of 3H-spiperone, 3H-N,N propylnorapomorphine and 3H-raclopride binding in the rat brain. AB - The stereospecific blockade by raclopride and FLB472 (the R enantiomer of raclopride) of the specific in vivo binding of [3H]-spiperone, [3H]-N,N propylnorapomorphine (NPA) and [3H]-raclopride was studied in seven brain regions (e.g., caudate nucleus, olfactory tubercle, septum, hippocampus, frontal cortex, substantia nigra, pituitary gland) of the male albino rat. The binding of all three ligands was dose-dependently blocked by raclopride and FLB472. The blockade by FLB472 occurred at doses 50-100 times higher than that obtained by raclopride. The maximal blockade by raclopride of [3H]-spiperone binding differed between brain areas. Thus, the largest blockade was obtained in the substantia nigra (95%), septum (90%), caudate nucleus (60%) and olfactory tubercle (60%), while the blockade of [3H]-spiperone binding in the frontal cortex and pituitary gland did not exceed 30% and 50%, respectively. In contrast to [3H]-spiperone, the in vivo binding of [3H]-NPA and [3H]-raclopride was prevented by 90-100% in all brain areas examined. Taken together, the present findings indicate that the in vivo binding of three radioactive ligands to a central dopamine D-2 receptor can be stereoselectively blocked by the enantiomers of raclopride. The findings suggest that, under in vivo conditions, [3H]-raclopride and [3H]-NPA may label a closely related receptor site. However only some of the [3H]-spiperone binding sites may be identical to the [3H]-raclopride binding sites. The findings indicate furthermore that the relative overlap of D-2 sites shared by [3H] spiperone and [3H]-raclopride may vary between brain regions. PMID- 2974880 TI - Inhibition of p-chlorophenylalanine-induced muricide behavior following TRH microinjection into the limbic structures in the rat. AB - The effect of central microinjections of thyrotropin releasing hormone (TRH) on muricide behavior was investigated in rats with chronically implanted cannulas into one of the limbic structures: the amygdala, the hippocampus or the nucleus accumbens. The rats were made aggressive by p-chlorophenylalanine administration. Saline injection did not inhibit muricide, whereas TRH (10 micrograms in 0.5 microliter per side) significantly suppressed mouse-killing reaction upon injection into each of three regions. It is suggested that the observed antiaggressive effect of TRH may involve stimulation of the central noradrenergic and/or serotonergic transmission. PMID- 2974881 TI - Suppression of conditioned avoidance by 8-OH-DPAT in the rat. AB - Rats were trained to perform an aversely motivated discriminative task in a shuttle-box. The conditioned avoidance response was selectively suppressed by 8 OH-DPAT in a dose-dependent manner (25-100 micrograms.kg-1). There were no statistically significant deficits in discriminative performance. The present results suggest antipsychotic-like properties of 8-OH-DPAT. PMID- 2974882 TI - Relationship between motor and cognitive disorders in Huntington's disease. AB - Akinesia and mental decline appear to be more appropriate criteria than hyperkinesia for the evaluation of the stage and progression of Huntington's disease (HD). In order to establish the relationship between motor and cognitive impairment in the disease, 20 non-demented HD patients were compared with 44 control subjects with respect to motor and cognitive performance. HD patients were significantly impaired in almost all cognitive functions in comparison with controls. Reaction time (RT) and movement time (MT) were considerably slower in HD patients when compared with controls and with patients with parkinsonism. Hyperkinesias did not correlate with cognitive impairment, but there was a good correlation between RT, MT and cognitive functions. Therefore, it seems that akinesia evaluated by RT and MT is an important sign in HD and proceeds at the same rate as mental decay. PMID- 2974883 TI - An attributional analysis of reactions to stigmas. AB - In two experiments, we examined the perceived controllability and stability of the causes of 10 stigmas. Guided by attribution theory, we also ascertained the affective reactions of pity and anger, helping judgments, and the efficacy of five intervention techniques. In the first study we found that physically based stigmas were perceived as onset-uncontrollable, and elicited pity, no anger, and judgments to help. On the other hand, mental-behavioral stigmas were perceived as onset-controllable, and elicited little pity, much anger, and judgments to neglect. In addition, physically based stigmas were perceived as stable, or irreversible, whereas mental-behavioral stigmas were generally considered unstable, or reversible. The perceived efficacy of disparate interventions was guided in part by beliefs about stigma stability. In the second study we manipulated perceptions of causal controllability. Attributional shifts resulted in changes in affective responses and behavioral judgments. However, attributional alteration was not equally possible for all the stigmas. PMID- 2974884 TI - Intragastric behavior and absorption kinetics of a normal and "floating" modified release capsule of isradipine under fasted and fed conditions. AB - From measurements of drug levels in both gastric juice and plasma, we investigated whether or not a prolonged gastric residence time (GRT) is responsible for the slow absorption kinetics of a "floating" modified-release (MR) capsule of isradipine [isopropyl methyl (+/-)-4-(4-benzofurazanyl)-1,4 dihydro-2,6-dimethyl-3,5- pyridinedicarboxylate], a lipophilic dihydropyridine calcium channel blocker. The effects of a "high-fat" breakfast on the intragastric behavior and absorption kinetics were also assessed. In an open crossover design, five healthy subjects ingested either a normal or MR capsule of isradipine under fasted conditions. Serial samples of gastric juice (obtained via an indwelling nasogastric tube) and plasma were collected up to 24 h after drug intake, and were analyzed for isradipine by GC and RIA methods, respectively. The pH and titratable acid, protein, and pepsin concentrations of the gastric juice samples were also determined. Four additional subjects were similarly studied after ingesting the capsules following a high-fat breakfast. Under fasted conditions, gastric juice drug levels of the normal and MR capsules indicated a median GRT of less than 1.5 h in both cases. Plasma levels indicated a rapid absorption for the normal capsule (less than 2 h), but a remarkably slow absorption for the MR capsule, lasting 24 h or more. Under fed conditions, gastric juice and plasma profiles of the normal capsule were similar to those for the fasted case. In contrast, the MR capsule had an increased GRT (approximately 2.4 to 4.8 h) that was associated with a delayed and more extensive intragastric drug release. The corresponding plasma profiles showed a rapid absorption phase which correlated closely with the intragastric release kinetics. The influence of a high-fat meal on the release kinetics of the MR capsule did not appear related to the intragastric pH, or acid, protein, or pepsin concentrations. From these results we conclude that: (1) a prolonged GRT is not responsible for the slow absorption achieved with a "floating" MR capsule; (2) the presence or absence of food, rather than buoyancy, is the principal determinant of the GRT of the MR capsule; (3) the release and absorption of a lipophilic drug from a "floating" MR capsule may be affected by intragastric interaction with the lipid phase of meal; and (4) the major portion of drug release from the MR capsule takes place in the colon, rather than in the stomach. PMID- 2974885 TI - Diagnosis of occupational dermatitis by general practitioners and employment medical advisors: a pilot study. AB - This pilot study examined how closely general practitioners and employment medical advisors agreed when jointly investigating occupational dermatitis. The criterion for admitting a patient to the study was dermatitis on one or both hands. This presented most commonly among men aged 41-60 years and women aged 21 40 years. There was some reluctance among general practitioners to take part in the study, probably owing to the medico-legal problems associated with this condition. General practitioners and employment medical advisors were equally likely to attribute a case of dermatitis to occupational factors if the patient was employed in industry; but if the patient was in nonindustrial employment, general practitioners were more likely to consider the possibility of an occupational origin than employment medical advisors. These differences suggest that before initiating a wider study more information is required concerning the trigger mechanisms for dermatitis on the hands in order to reduce the diagnostic variability. PMID- 2974886 TI - Duplicated region of the mouse genome containing a cytoplasmic gamma-actin processed pseudogene associated with long interspersed repetitive elements. AB - The structures of two cloned recombinants of bacteriophage lambda and mouse genomic DNA (lambda mA14 and lambda mA36) were compared by electron microscopic analysis of various heteroduplex DNAs, restriction endonuclease mapping and nucleotide sequence determination. Each clone was shown to be derived from a distinct region of the mouse genome, but the two exhibited structural similarity over a region of at least 11,000 bases which included a cytoskeletal gamma-actin processed pseudogene of approximately 1800 bases. It is concluded that the two genomic regions were derived from a common ancestral region by duplication or amplification. The homologous regions of the two clones contained members of the long interspersed repetitive L1Md (long interspersed repeated sequence 1 of Mus domesticus) family lying in opposite orientation to one another, so that single stranded DNA from the clones could form intra-molecular heteroduplexes. The complete nucleotide sequences of three L1Md members in lambda mA14 were determined. The longest of these (L1Md-14LH) had inserted into the gamma-actin processed pseudogene and, although it contained internal deletions, appeared to possess intact 5' and 3' ends. A second L1Md member (L1Md-14RH1) also appeared to have an intact 5' end but had lost most of its 3' portion, and a third member (L1Md-14RH2) was an internal fragment. The repeated sequence at the 5' ends of L1Md-14LH and L1Md-14RH1 showed these to be members of the L1Md-A family. PMID- 2974887 TI - Biochemical basis of the temperature-inducible constitutive protease activity of the RecA441 protein of Escherichia coli. AB - We compared the biochemical properties of the RecA441 protein to those of the wild-type RecA protein in an effort to account for the constitutive protease activity observed in recA441 strains. The two RecA proteins have similar properties in the absence of single-stranded DNA binding protein (SSB protein), and the differences that do exist shed little light on the temperature-inducible phenotype observed in recA441 strains. In contrast, several biochemical differences are apparent when the two proteins are compared in the presence of SSB protein, and these are conducive to a hypothesis that explains the temperature-sensitive behavior observed in these strains. We find that both the single-stranded DNA (ssDNA)-dependent ATPase and LexA-protease activities of RecA441 protein are more resistant to inhibition by SSB protein than are the activities of the wild-type protein. Additionally, the RecA441 protein is more capable of using ssDNA that has been precoated with SSB protein as a substrate for ATPase and protease activities, implying that RecA441 protein is more proficient at displacing SSB protein from ssDNA. The enhanced SSB protein displacement ability of the RecA441 protein is dependent on elevated temperature. These observations are consistent with the hypothesis that the RecA441 protein competes more efficiently with SSB protein for limited ssDNA sites and can be activated to cleave repressors at elevated temperature by displacing SSB protein from the limited ssDNA that occurs naturally in Escherichia coli. Neither the ssDNA binding characteristics of the RecA441 protein nor the rate at which it transfers from one DNA molecule to another provides an explanation for its enhanced activities, leading us to conclude that kinetics of RecA441 protein association with DNA may be responsible for the properties of the RecA441 protein. PMID- 2974888 TI - Regional changes in myocyte size during the reversal of thyroid-induced cardiac hypertrophy. AB - Regional changes in cardiac myocyte size resulting from thyroid treatment, and the subsequent reversal of hypertrophy following removal of the thyrotoxic stimulus, were examined. Isolated myocytes were prepared from weight-matched controls and rats treated with desiccated thyroid hormone for 10 weeks. Cells were collected from the right and left ventricles. Cell volume was determined with a Coulter Channelyzer system. Cell length was measured directly using a phase microscope. Myocyte cross-sectional area was calculated from cell volume/length. After 10 weeks thyroid treatment, cell volume was increased in all regions, especially the right ventricle and the epimyocardium of the left ventricle (P less than 0.01). Although cell length was increased in all regions (N.S.), most of the myocyte hypertrophy was due to an increase in cross-sectional area, especially in the right ventricle (P less than 0.01) and epimyocardium of the left ventricle (N.S.). By 2 weeks post-treatment, a substantial regression of myocyte hypertrophy had occurred. However, heart weight was still significantly larger than control (P less than 0.001), due to an incomplete regression of myocyte volume. This study indicates that thyroid hormones stimulate myocyte hypertrophy by increasing both cross-sectional area and cell length. The response is more pronounced in right ventricles than in left ventricles. Within the left ventricle, epimyocardial cells enlarge the most. At 2 weeks post-treatment, cardiac hypertrophy is still present and myocytes have not completely returned to a normal size. PMID- 2974889 TI - The impact of diabetes on survival following myocardial infarction in men vs women. The Framingham Study. AB - The impact of diabetes on recurrent myocardial infarction (MI) and fatal coronary heart disease was examined in survivors of an initial MI using 34-year follow-up data in the Framingham Study. Among nondiabetic patients, the risk of fatal coronary heart disease was significantly lower in women compared with men (relative risk, 0.6). In the presence of diabetes, however, the risk of recurrent MI in women was twice the risk in men. In addition, the effect of diabetes doubled the risk of recurrent MI in women (relative risk, 2.1) but had an insignificant effect in men. Increased susceptibility to cardiac failure among diabetic women was an important factor in determining survivorship. Women with diabetes developed cardiac failure four times more often (16%) than women without diabetes (3.8%). Furthermore, when cardiac failure developed, 25% of diabetic women experienced a recurrent MI or fatal coronary event, more than doubling the rate when diabetes was absent. We conclude that in the diabetic patient who survives an MI, cardiac failure is a common occurrence, warranting early detection and vigorous management in periods of convalescence and follow-up. In addition, when cardiac failure appears, control of diabetes assumes added importance, particularly in women, where its effect on survivorship is considerable. PMID- 2974891 TI - New electrocardiographic criteria for diagnosing right ventricular hypertrophy in mitral stenosis--comparison with the Bonner's and Mortara's criteria. AB - New criteria for the diagnosis of right ventricular hypertrophy (RVH) using a point scoring system were developed after analyzing standard 12-lead ECGs in 155 patients with mitral stenosis verified by cardiac catheterization and 155 age and sex-matched apparently normal healthy subjects. ECGs were evaluated to identify criteria that provided maximum sensitivity and at least a 95% specificity: (1) the R wave magnitude in V1 had to be greater than 0.7 mV; (2) the S wave magnitude in V6 had to be greater than 0.3 mV; (3) the S wave magnitude in V1 less than 0.5 mV; (4) the R wave magnitude in V1 plus the S wave magnitude in V6 minus the S wave magnitude in V1 must be greater than 0 mV; and (5) the degree of frontal QRS axis had to be greater than 90 degrees. Application of these criteria achieved 25% (39 of 155) sensitivity in patients with RVH, significantly better (p less than 0.01) than the 17% sensitivity (17 of 155) of Bonner's criteria for the automated ECG. The specificity of the proposed criteria was 98% (152 of 155), significantly better (p less than 0.01) than the 90% specificity (139 of 155) of Mortara's criteria. Thus, it was evident that the accuracy of the proposed criteria was the highest among those criteria used in a point scoring system including the currently used automated ECG criteria for the diagnosis of RVH. PMID- 2974890 TI - Effects of alpha human atrial natriuretic polypeptide on the systemic hemodynamics and coronary circulation in patients with ischemic heart disease. AB - Alpha human atrial natriuretic polypeptide (alpha-hANP) was intravenously infused into 7 patients with ischemic heart disease who had almost normal cardiac function at a rate of 0.025 micrograms/kg/min for 15 min. During infusion of alpha-hANP, left ventricular (LV) systolic pressure decreased from 144 +/- 19 (SD) to 129 +/- 22 mmHg (p less than 0.01), LV end diastolic pressure (EDP) from 15 +/- 5 to 13 +/- 4 mmHg (p less than 0.05), mean aortic pressure from 102 +/- 14 to 91 +/- 14 mmHg (p less than 0.01), time constant of LV pressure fall (T) from 100 +/- 15 to 88 +/- 13 msec (p less than 0.05), systemic vascular resistance (SVR) from 1711 +/- 206 to 1424 +/- 340 dynes.sec.cm-5 (p less than 0.05) and coronary vascular resistance (CVR) from 8.5 +/- 1.2 to 7.4 +/- 1.3 x 10(4) dynes.sec.cm-5 (p less than 0.05). There was a linear correlation between percent changes in SVR and those of CVR (r = 0.92, p less than 0.01), and the fall in CVR was approximately 68% of that in SVR. Increases occurred in heart rate from 63 +/- 7 to 66 +/- 8 beats/min (p less than 0.05), LV dp/dt from 1558 +/- 266 to 1627 +/- 238 mmHg/sec (p less than 0.05), LV dp/dt/p from 22.9 +/- 3.2 to 25.6 +/- 3.7/sec (p less than 0.01), and myocardial oxygen consumption (from 7.9 +/- 2.4 to 9.8 +/- 2.1 ml/min, p less than 0.05), while mean right atrial and mean pulmonary arterial pressures and pulmonary vascular resistance were unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974892 TI - Isolation and characterization of two beta-type cardiac myosin in the canine atrium. AB - Recently, we demonstrated that more beta-type myosin heavy chain (HC) was expressed in the overloaded atrium, and that there were 2 structurally different beta-type myosin heavy chains in the bovine heart. To determine the existence of the 2 beta-type HC in other animals and to clarify the characteristics of these beta-type HCs, we produced tricuspid regurgitation and pulmonary stenosis in the canine heart, and performed an immunological study using 3 monoclonal antibodies, 2 beta-type specific antibodies (HMC14 and 50) and 1 alpha-type specific antibody (CMA19). In an immunohistochemical study, serial cryostat sections revealed that some myofibers reacted with HMC50 (HC beta 2), but almost no fibers were labeled with HMC14 in the normal atrium. However, in overloaded atria, not only HC beta 2 but the HC, reacted with HMC14 (HC beta 1). By affinity chromatography, HC beta 2 was fractionated from normal atrial myosin using HMC50 and HC beta 1 was fractionated from overloaded atrial myosin using HMC14. These 2 HC beta's were subjected to digestion by alpha-chymotrypsin, staphylococcus aureus V8 protease, and cyanogen bromide, and proved to have different peptide fragments. In respect to enzymatic properties, the Ca2+-activated ATPase activities of HC beta 1 and beta 2 were almost the same but lower than that of HC alpha. We concluded that the isozymic transition of HC alpha to HC beta in the atrium was experimentally induced by hemodynamic overload and that HC beta 1, which was hardly recognized in the normal atrium but highly induced by overload, was structurally different from HC beta 2, as expressed in the normal atrium. PMID- 2974893 TI - Abnormal polyamine metabolism in hypertensive cardiac hypertrophy. AB - In order to assess myocardial hypertrophic activity during the process of hypertensive cardiac hypertrophy in the presence and absence of treatment with anti-hypertensive agents, we analyzed myocardial polyamine concentrations in spontaneous hypertensive (SHR) rats and control rats of Wistar Kyoto (WKY) strain. The anti-hypertensive agents studied were diltiazem, hydralazine and captopril, each of which was administered for 5 weeks. In comparison with WKY rats, SHR rats showed elevated blood pressure and enlarged hearts with higher myocardial spermidine concentration. Although blood pressure was lowered in the diltiazem-treated SHR rats, heart weight and myocardial spermidine concentration increased as in untreated SHR rats. In the hydralazine-treated group increases in both blood pressure and myocardial spermidine concentration were suppressed, while an increase in heart weight was not. In the captopril-treated group, increases in blood pressure, heart weight and spermidine concentration were all suppressed. Since spermidine level appears to be a sensitive indicator of hypertrophic activity in the heart, this study suggests that captopril exerts an inhibitory effect on hypertensive cardiac hypertrophy whereas diltiazem does not. It also suggests that hypertrophy may reach a certain plateau level earlier in the hydralazine-treated animals than in others. PMID- 2974894 TI - [Hemostatic tests for the diagnosis of thrombosis. Activated coagulation and fibrinolytic factors. Detection of the protease/inhibitor complex]. PMID- 2974895 TI - Studies on the Fc receptors and masking substances on mouse and rat myeloma cells. AB - Mouse and rat myeloma cell lines showed little or no rosette formation with sheep erythrocytes (SRBC) coated with rabbit IgG (rabbit EA) but showed marked rosette formation after treatment with trypsin, pronase or neuraminidase. These cell lines showed no rosette formation with SRBC coated with mouse IgG (mouse EA): treatment with trypsin enabled the detection of rosettes among the mouse myeloma cell lines but not by the rat myeloma cells. The F(ab')2 fragment of the anti-Fc receptor II antibody blocked the formation of rosettes with rabbit EA by mouse myeloma cell lines after treatment with trypsin. Aggregated mouse IgG1 and IgG2b subclasses strongly inhibited the formation of rosettes with rabbit EA, whereas aggregated mouse IgG2a showed a marginal inhibitory effect. A large amount of mouse IgG2a, however, caused significant inhibition. Our results also revealed that aggregated mouse IgG could bind to the rat myeloma cell line. The Fc rosette forming abilities of the enzyme-treated mouse and rat myeloma cells became reduced after cultivation both in the presence and absence of FCS but not after cultivation in the presence of cycloheximide, suggesting that cell surface substances, which may be glycoprotein that incompletely mask Fc receptors, are produced by myeloma cells. PMID- 2974897 TI - [A nursing scene filled with childrens' laughter--Kita Center for Handicapped Children in Tokyo]. PMID- 2974898 TI - [Self-care and guidance of patients: nursing of patients with chronic rheumatoid arthritis. Self-care and family functions--assistance toward independence of handicapped patients]. PMID- 2974896 TI - [Study of plasma beta thromboglobulin level induced by heparin in diabetic patients]. PMID- 2974899 TI - [A case report of rupture of the descending thoracic aorta]. PMID- 2974900 TI - [Balloon dilatation angioplasty of stenotic pulmonary artery simultaneously with a Blalock-Taussig shunt in extremely tetralogy of Fallot]. PMID- 2974901 TI - [Suppression of immune response in mice treated with live and killed BCG]. PMID- 2974902 TI - [Possibilities of using magnetometry methods of examination in cardiology]. PMID- 2974903 TI - [Automated selection of optimal parameters of precordial mapping in the diagnosis of right-ventricular hypertrophy]. AB - New diagnostic parameters, improving the specificity and sensitivity of electrocardiographic diagnosis of right-ventricular hypertrophy, have been identified on the basis of an automated analysis of precordial maps in 56 patients with isolated right-ventricular hypertrophy and 71 normal subjects. PMID- 2974904 TI - [Newly developed stenocardia: functional activity of thrombocytes]. AB - Platelet function (aggregation, circulating platelet aggregates, platelet factor 4, beta-thromboglobulin, thromboxane B2, 6-keto-PGF1 alpha, platelet count) was evaluated in patients with angina of new onset, as compared to individuals without coronary heart disease, and patients with angina of long standing (28 +/- 4.6 months). Some of the patients with angina of new onset were examined repeatedly 6 to 12 months after the onset of angina. Platelet activity was shown to be significantly higher in patients with stable angina of new onset, as compared to patients with lasting angina, where ADP and serotonin were used as inductors. Platelet aggregation, induced by platelet activation factor and collagen, was similar in all groups. Repeated investigation 6 to 12 months after the first anginal attacks demonstrated that most of platelet functional parameters declined or tended to decline. PMID- 2974906 TI - [Difficulties and errors in the ambulatory diagnosis of soft tissue sarcomas]. PMID- 2974905 TI - Atrial natriuretic peptide and the renal response to hypervolemia in nephrotic humans. AB - To elucidate the abnormality of body fluid homeostasis that attends the nephrotic syndrome, we compared the atrial hormonal and renal excretory and vasomotor responses to water immersion of nephrotic patients (N = 10) with those of healthy controls (N = 9). Nephrotics exhibited depressed baseline levels of atrial natriuretic peptide (ANP, P less than 0.05) and lower rates of urine flow and sodium excretion (P less than 0.01). Although immersion-induced hypervolemia increased plasma ANP to equivalent levels (75 +/- 19 vs. 60 +/- 6 pg/ml), the disparity in corresponding urinary flow (5 +/- 1 vs. 13 +/- 2 ml/min, P less than 0.01) and sodium excretion (171 +/- 42 vs. 540 +/- 65 muEq/min, P less than 0.01) grew larger. In contrast, immersion caused an equivalent reduction of renal vascular resistance by 16 and 17%, respectively (P less than 0.01). Despite higher renal plasma flow and lower oncotic pressure of plasma, the glomerular filtration rate remained constant during immersion in both groups. Similar constancy of fractional clearances of dextrans of graded size suggests that immersion may have lowered the glomerular transcapillary hydraulic pressure difference (delta P). We conclude that renal vasomotor responsiveness to hypervolemia is preserved in nephrotics, but that the mediatory role of ANP in this response is uncertain. By contrast, diminished responsiveness of the distal nephron to the natriuretic action of endogenous ANP could contribute to edema formation in the nephrotic syndrome. PMID- 2974907 TI - [Cancer of the breast in men]. PMID- 2974908 TI - [Active detection of preneoplastic and neoplastic diseases at an endoscopic center]. PMID- 2974909 TI - [The dose of intracoronary urokinase infusion in acute myocardial infarct]. AB - In 11 patients with acute myocardial infarction (mean age 52 +/- 6 years) coronary angiography was performed 4.5 +/- 3.6 h after the onset of symptoms. The infarct-related artery was in 7 cases the right coronary artery and in 4 cases the left anterior descending branch of the left coronary artery. The infarct related artery showed total or subtotal occlusion and no perfusion (Thrombolysis in Myocardial Infarction trial (TIMI) grade 0 or 1). In 7 cases Urokinase was infused intracoronarily at a dosage of 250,000 IU over 30 min, but in only 1 case partial reperfusion was achieved. However, all patients treated with 500,000 IU Urokinase over 30 min or 1 Mill IU over 60 min had successful reperfusion (TIMI grade 2 or 3). Thus, it appears that 500,000 IU up to 1 Mill IU of Urokinase over a period of 30 to 60 min is adequate for intracoronary thrombolysis in patients with acute myocardial infarction. PMID- 2974910 TI - [Enjoyment of basketball in wheelchairs]. PMID- 2974911 TI - Design of a continuous-wave Doppler ultrasonic flowmeter for perivascular application. Part 1. Probe design. PMID- 2974912 TI - Difference in second-formant transitions between aspirated and unaspirated stop consonants preceding [a]. PMID- 2974913 TI - Atrial natriuretic peptides cleaved by endopeptidase are inactive in conscious spontaneously hypertensive rats. AB - The dose-related natriuretic and depressor responses to atrial natriuretic peptides (ANP) 99-126, 103-126 and 103-123 were determined in unanesthetized spontaneously hypertensive rats (SHR) and were compared to the activities of their Cys105-Phe106 ring-opened metabolites. These metabolites were previously identified as the major initial products formed by incubation of the intact peptides with neutral endopeptidase (NEP). The areas over the curves (AOC) of the depressor responses to the intact peptides were dose-related and, at 30 nmole/kg, iv were greatest for ANP 99-126 and 103-126 (833 +/- 241 and 1157 +/- 221 mm Hg x min). Thirty nmole/kg of ANP 103-123, a possible product of NEP cleavage of ANP 103-126, produced a lesser AOC (442 +/- 152 mm Hg x min) than did either of the longer peptides. The AOC responses to 100 nmole/kg of the ring-opened metabolites of ANP 99-126, 103-126 and 103-123 (105 +/- 80, 153 +/- 43 and 148 +/- 64 mm Hg x min) were not significantly different from the effect of vehicle treatment (84 +/ 23 mm Hg x min). Although the natriuretic responses to increasing doses of the intact peptides did not occur in a linear fashion, sodium excretion was maximally elevated by 24 +/- 4, 16 +/- 3 and 10 +/- 3 microEq/kg/min by 3 nmole/kg of ANP 99-126, 30 nmole/kg of ANP 103-126 and 10 nmole/kg of ANP 103-123, respectively. In contrast, the natriuretic responses to 100 nmole/kg of the ring-opened metabolites of ANP 99-126, 103-126 and 103-123 (1 +/- 0, 5 +/- 2 and 2 +/- 1 microEq/kg/min, respectively) were not significantly different from the response to vehicle treatment (3 +/- 1 microEq/kg/min). In conclusion, three ring-opened products of NEP cleavage of ANP 99-126, 103-126 and 103-123 were inactive in conscious SHR. PMID- 2974914 TI - [Coexistence of cervico-brachial neuralgia and backache among textile industry workers]. AB - The paper has been aimed at searching for the correlation between neck and arm pain and low back pain to test the rate of concomitance of both disorders and the order of their appearance. The studies have been carried out in two big light industry plants in Lodz. The coexistence of the pain syndromes has been found to be frequent, low back pain usually developing first. It seems that the occurrence of one of those disorders disturbs the functioning of the whole vertebral column and thus predisposes to the development of the other one. PMID- 2974915 TI - [Atrial natriuretic peptide]. PMID- 2974916 TI - Enzymes of glucose metabolism in cultured human gliomas: neoplasia is accompanied by altered hexokinase, phosphofructokinase, and glucose-6-phosphate dehydrogenase levels. AB - The enzymes of glycolysis and selected enzymes of the pentose phosphate pathways were measured by fluorometric methods in extracts prepared from cultures of normal cortical human astrocytes and from cultures derived from low-grade (II) or high-grade (IV) gliomas. The hexokinase and phosphofructokinase levels of the low grade glioma-derived line were not significantly different from those of the normal astrocyte cultures. However, the activities of hexokinase and phosphofructokinase were consistently and significantly increased in the high grade glioma-derived lines. The activity of glucose-6-phosphate dehydrogenase was significantly decreased in all glioma-derived lines and by more than 90% in the high-grade-derived lines. Other enzymes of the glycolytic pathway were not significantly different from those of normal astrocytes, or they showed a variation inconsistently related to the neoplastic state. Glucose flux is not apparently regulated to a significant degree of hexokinase in glioma-derived lines, since the measured Vmax values are in substantial excess over the measured flux rates. Reversible binding of hexokinase to the particulate fraction was observed in both the normal astrocytes cultures and the high-grade glioma-derived lines. A twofold displacement of particulate hexokinase by ATP, ADP, 1-O methylglucose, sorbitol-6-phosphate, and dibutyryl cyclic AMP was observed in the high-grade glioma-derived lines. The degree of displacement by various agents and the basal ratio of free/bound was not significantly different between the transformers and the nontransformants. The hexokinase from both the gliomas and the normal astrocytes was noncompetitively inhibited by the glucose analogue 2 deoxy-d-glucose. Phosphofructokinase activity is close to the observed glucose flux rates in both the normal astrocyte and the glioma-derived cultures. The phosphofructokinase activity of normal astrocytes is activated twofold or more by ADP, AMP, fructose-2,6-diphosphate, and Pi. However, these same ligands activate phosphofructokinase by less than twofold in a typical high-grade glioma-derived line. ATP, dibutyryl cyclic AMP, and citrate inhibit glioma and normal astrocytic phosphofructokinase, but the magnitude of the inhibition is much less than in the glioma-derived lines. PMID- 2974918 TI - High-risk dentistry (AIDS and hepatitis): role of the rubber dam. PMID- 2974917 TI - On forecasting mortality. AB - Official forecasts of mortality made by the U.S. Office of the Actuary throughout this century have consistently underestimated observed mortality declines. This is due, in part, to their reliance on the static extrapolation of past trends, an atheoretical statistical method that pays scant attention to the behavioral, medical, and social factors contributing to mortality change. A "multiple cause delay model" more realistically portrays the effects on mortality of the presence of more favorable risk factors at the population level. Such revised assumptions produce large increases in forecasts of the size of the elderly population, and have a dramatic impact on related estimates of population morbidity, disability, and health care costs. PMID- 2974919 TI - Increased amount of a 25-kilodalton phosphoprotein after v-mos transfection of CHO cells. AB - We transfected Chinese hamster ovary (CHO) cells with a cloned v-mos gene (pHT25). The mos family of oncogenes has previously been shown to have serine threonine kinase activity. This kinase activity may be required for oncogenic transformation, although its exact biological role is unknown. We found that the transfected cells had an altered morphology, a slower doubling time, and an apparent increase in the amount of a 25-kilodalton (kDa) phosphoprotein that appeared to be of low abundance. Transfection of CHO cells with a cloned temperature-sensitive mos gene (ts159) led to isolation of a cell line that showed the presence of the 25-kDa phosphoprotein at the permissive but not at the nonpermissive temperature, suggesting a direct relationship between mos activity and the presence of this phosphoprotein. The characteristics of altered morphology and depressed growth rate were reminiscent of changes seen after the activation of the cyclic AMP-dependent protein kinase (PKA) in CHO cells. However, PKA activation did not stimulate phosphorylation of this 25-kDa protein, nor was there a change in total PKA activity in these cells. We suggest that the increased presence of the 25-kDa phosphoprotein is a consequence of the v-mos transfection and that it may be involved in the change of morphology and growth rate seen in the CHO cells. Phosphorylation of this protein may be a useful marker of mos and have some functional importance in the transformation of cells by the v-mos oncogene. PMID- 2974920 TI - Loop I of U1 small nuclear RNA is the only essential RNA sequence for binding of specific U1 small nuclear ribonucleoprotein particle proteins. AB - The binding of the U1 small nuclear ribonucleoprotein (snRNP)-specific proteins C, A, and 70K to U1 small nuclear RNA (snRNA) was analyzed. Assembly of U1 snRNAs from bean and soybean and a set of mutant Xenopus U1 snRNAs into U1 snRNPs in Xenopus egg extracts was studied. The ability to bind proteins was analyzed by immunoprecipitation with monospecific antibodies and by a protein-sequestering assay. The only sequence essential for binding of the U1-specific proteins was the conserved loop sequence in the 5' hairpin of U1. Further analysis suggested that protein C binds directly to the loop and that the assembly of proteins A and 70K into the RNP requires mainly protein-protein interactions. Protein C apparently recognizes a specific RNA sequence rather than a secondary structural element in the RNA. PMID- 2974921 TI - Concentration dependence of transcriptional transactivation in inducible E1A containing human cells. AB - The adenovirus E1A proteins are essential for the normal temporal activation of transcription from every other adenoviral early promoter. High-level E1A expression in the absence of viral infection would facilitate biochemical studies of E1A-mediated transactivation. Toward this end, we introduced the adenovirus type 2 E1A gene under the control of the murine mammary tumor virus promoter into HeLa cells. Uninduced cells expressed little or no detectable E1A mRNA. Upon induction, mRNA levels accumulated to about 50% of the level observed in 293 cells. The level of E1A expression in these cells could be controlled by varying the concentration of the inducing glucocorticoid. Under these conditions of varying E1A concentrations, it was observed that activation of the E2, E3, and E4 promoters of H5dl312 initiated at the same E1A concentration and that transcription from each promoter increased as the E1A concentration increased. These results indicate that E1A-mediated transactivation is proportional to the concentration of E1A protein. E1A-dependent transcriptional stimulation of the E4 promoter was reproduced in an in vitro transcription system, demonstrating that expression of only the E1A proteins was sufficient to increase the transcriptional activity of nuclear extracts. PMID- 2974922 TI - Organization of the murine Mx gene and characterization of its interferon- and virus-inducible promoter. AB - Specific resistance of Mx+ mice to influenza virus is due to the interferon (IFN) induced protein Mx. The Mx gene consists of 14 exons that are spread over at least 55 kilobase pairs of DNA. Surprisingly, the Mx gene promoter is induced as efficiently by Newcastle disease virus as it is by IFN. The 5' boundary of the region required for maximal induction by both IFN and Newcastle disease virus is located about 140 base pairs upstream of the cap site. This region contains five elements of the type GAAANN, which occurs in all IFN- and virus-inducible promoters. The consensus sequence purine-GAAAN(N/-)GAAA(C/G)-pyrimidine is found in all IFN-inducible promoters. PMID- 2974923 TI - Growth factor induction by the adenovirus type 5 E1A 12S protein is required for immortalization of primary epithelial cells. AB - The 12S protein encoded by the adenovirus E1A region induces cellular DNA synthesis in and proliferation and immortalization of primary rat epithelial cells in the presence or absence of serum. It also induces the production of a growth factor(s) that stimulates epithelial cell proliferation. We have undertaken a mutational analysis of the 12S gene to determine the sequences required for these functions. We found that a region near the C-terminus of the 12S protein was required for growth factor induction. No activities have been defined previously for this region. Furthermore, we show that growth factor production was necessary for epithelial cells to survive past their normal life span in culture and to become immortalized. The ability to induce growth factor production required prior expression of E1A activities encoded by the N-terminus of the 12S protein, including activation of quiescent cells into the cell cycle, and an unknown activity that required expression of the first 13 amino acids of the gene. In addition, examination of the subcellular localization of mutant 12S polypeptides suggested new regions that affect the nuclear localization of E1A proteins. PMID- 2974924 TI - Step-arrest mutants of FLP recombinase: implications for the catalytic mechanism of DNA recombination. AB - The site-specific recombinase (FLP) encoded by the yeast plasmid 2 micron circle belongs to the integrase (of phage lambda) family of recombinases. The sparse homology within the members of this family contrasts with the invariance of three residues, His-396, Arg-399, and Tyr-433 (the numbers correspond to the family alignment positions), among them. We report here results on substrate recognition and catalysis by FLP proteins altered at these residues. Mutations of the conserved His and Tyr that aborted the reaction at specific steps of catalysis permitted genetic dissection of the possible biochemical steps of recombination. We provide indirect evidence that recombination by FLP proceeds through a Holliday junction intermediate. PMID- 2974925 TI - Presence of the adenovirus E1A-like activity in preimplantation stage mouse embryos. AB - The presence of the adenovirus E1A-like activity in embryonal carcinoma stem cells has been reported. We now show that preimplantation stage mouse embryonic cells allow transcription of the E1A-dependent E2A gene when infected with E1A deleted mutant dl312, indicating the presence of the E1A-like activity in morulae and blastocysts. Moreover, such activity seems to decrease or disappear at about the time of implantation. PMID- 2974926 TI - Human calpactin II (lipocortin I) messenger ribonucleic acid is not induced by glucocorticoids. AB - Studies were carried out to examine the effect of glucocorticoids on human calpactin II (lipocortin I) mRNA expression. A cRNA probe for human calpactin II (hCPII) was used in a solution hybridization assay to study the effect of dexamethasone on hCPII mRNA levels in human skin fibroblasts, peripheral lymphocytes, pulmonary alveolar macrophages, and HeLa S3 cells. As a positive control, human metallothionein II (hMTII) mRNA levels were measured since hMTII is known to be regulated by glucocorticoids and heavy metals, both of which induce transcriptional activity of the gene. Dexamethasone treatment of these human cell types caused a dose-dependent increase in hMTII mRNA levels, whereas no effect on hCPII mRNA levels was observed. These findings were confirmed in time course studies, where 10(-6) M dexamethasone treatment caused a maximal 2- to 5-fold increase in hMTII mRNA levels after 6-8 h of treatment but no increase in hCPII mRNA levels was observed at any time point up to 24 h. A human glucocorticoid sensitive lymphoid cell line, CEM C7, and a glucocorticoid resistant mutant, ICR-27, isolated from CEM C7, were included in order to confirm the requirement of a functional glucocorticoid receptor (GR) in the induction of glucocorticoid-regulated genes. Dexamethasone (10(-6) M) induced hMTII but not hCPII mRNA in CEM C7 cells, whereas neither hMTII nor hCPII mRNA was induced in ICR-27 cells. In conclusion, our data suggest that glucocorticoids do not induce calpactin II (lipocortin I) mRNA in the human cell types studied. PMID- 2974928 TI - Somatosensory phenomena in Huntington's disease. AB - Sensory symptoms are generally not associated with Huntington's disease (HD). We describe two patients with HD who had painful somatosensory symptoms. One patient also had auditory hallucinations. No other cause was found for these symptoms. Both patients also had significant depression and one patient committed suicide. Somatosensory symptoms may be a marker for depression in HD. PMID- 2974927 TI - Correlation between facial involuntary movements and abnormalities of blink and corneal reflexes in Huntington's chorea. AB - Blink and corneal reflexes were studied in 11 patients with Huntington's chorea and the results compared with the severity of the disease. The latency of the R2 component of the blink reflex was delayed and the duration of R2 and of the corneal reflex (CR) prolonged. A greater habituation of the R2 component was found in the patients with involuntary movements in the face, and in some patients a long-lasting depression of R2 was present. A correlation was found between: (a) severity of involuntary face movements and R2 and CR latency and (b) severity of involuntary movements in the neck and latency of R2. PMID- 2974929 TI - Neuroendocrinological function in Alzheimer's disease. AB - The neuroendocrine function is regulated by several neurotransmitters (acetylcholine, dopamine, somatostatin and noradrenaline) known to be reduced in brains of patients with Alzheimer's disease (AD). Moreover, the hypothalamus also has pathological changes. In spite of these findings suggesting neuroendocrine dysfunctions, this function has seldom been investigated in AD patients so far. We have compared patients with clinically 'probable' AD of mild-to-moderate severity with nondemented age- and sex-matched controls. Plasma levels of prolactin (PRL), growth hormone (GH) and thyroid-stimulating hormone (TSH) were measured by commercially available radioimmunoassays (RIA) before and after stimulation with metoclopramide, l-dopa or thyrotropin-releasing hormone. Basal plasma levels of beta-endorphin and beta-lipotropin were measured by RIA after high-performances liquid chromatography. Basal and stimulated plasma levels of PRL, GH, TSH and beta-lipotropin were similar in the two groups. Basal lamina levels of beta-endorphin were significantly higher in the patient group. Of doubtful clinical importance, this might be attributed to decreased tuberoinfundibular dopaminergic activity and has also been seen in patients with Parkinson's disease. PMID- 2974930 TI - [Diltiazem in unstable angina: a therapeutic proposal]. PMID- 2974932 TI - [Percutaneous transluminal angioplasty in stenosis following a surgical portosystemic shunt]. PMID- 2974931 TI - [Antiaggregant effect of slow-release dipyridamole in acute myocardial infarct]. PMID- 2974933 TI - [The OKT4/OKT8 ratio in benign monoclonal gammopathies]. AB - The percentages of OKT 4 helper and OKT 8 suppressor lymphocytes were studied in 58 patients in whom a monoclonal protein was accidentally identified in the absence of any clinical signs of multiple myeloma, macroglobulinaemia, amyloidosis or lymphoma. The patients were aged 32-63 and were followed-up for 6 years. The F test used in this study revealed a statistically significant difference between the healthy controls and the patients with BMG in one of the parameters examined: OKT 8. This produced a statistically significant alteration in the OKT 4/OKT 8 ratio. PMID- 2974934 TI - [Comparison of protective effects induced by the ultrasonic mist of nicardipine + fenoterol versus nicardipine + fenoterol + ipratropium bromide]. AB - A comparison is presented of the protective effect of ultrasound mist induced by nicardipine + fenoterol and nicardipine + fenoterol + ipratropium bromide in a group of 11 patients with chronic bronchial asthma in a phase of relative clinical stability. The data collected show that the three-drug combination was more effective than the use of nicardipine and fenoterol. PMID- 2974935 TI - [Treatment of reflex neurodystrophic syndromes of lumbar osteochondrosis using hydroxycobalamin injections]. PMID- 2974936 TI - [Agricultural injuries in viticulture and their prevention]. PMID- 2974937 TI - [Prevention of road and transportation injuries of children]. PMID- 2974938 TI - Arrhythmias during combined procaterol-aminophylline treatment. A review of the literature and a personal series with dynamic electrocardiography. PMID- 2974939 TI - Use of fetal streptozotocin injection to determine the role of normal levels of fetal insulin in regulating uteroplacental and umbilical glucose exchange. AB - The present study was performed to determine the role of the normal fetal concentration of insulin in regulating placental-fetal glucose exchange. Fetal insulin deficiency was produced by streptozotocin injection into near term fetal sheep, and the effects of this insulin deficiency on net uteroplacental glucose uptake and net umbilical glucose uptake were measured. Each fetus received two or three doses of streptozotocin, 100 mg.kg-1.dose-1, given on separate days. This dosage of streptozotocin produced a 97.6% reduction in fetal pancreatic insulin content, a fall in fetal plasma insulin concentration (21 +/- 2 to 10 +/- 1 microU.ml-1), a rise in fetal plasma glucagon concentration (57 +/- 4 to 114 +/- 19 pg.ml-1), a rise in fetal blood glucose concentration (20.4 +/- 0.9 to 33.4 +/ 4.4 mg.dl-1), and a failure of insulin secretion in response to glucose infusion. Fetal blood oxygen content and umbilical oxygen uptake were normal and did not change during the entire study. Umbilical glucose uptake was reduced by 66% (5.98 +/- 0.38 to 2.02 +/- 1.31 mg.min-1.kg-1) after the streptozotocin induced hypoinsulinemia and hyperglycemia but was returned to the control level by an insulin infusion into the fetus that reestablished the control maternal to fetal glucose concentration gradient. Net uteroplacental glucose uptake (consumption) did not change throughout the study. Because glucose concentration and umbilical glucose uptake could be normalized by an insulin infusion, it is unlikely that direct or toxic effects of streptozotocin on fetal or placental glucose metabolism were primarily responsible for the hyperglycemia and the reduced rate of umbilical glucose uptake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974940 TI - Fetal atrial natriuretic factor and arginine vasopressin responses to hyperosmolality and hypervolemia. AB - Atrial natriuretic factor (ANF) is a class of diuretic and natriuretic peptides secreted by mammalian cardiac atria. Although basal plasma ANF levels in the ovine fetus are elevated relative to the adult, fetal secretion of ANF increases in response to intravascular isotonic saline infusion. Recent in vitro and in vivo studies indicate that ANF secretion also may be stimulated by increased plasma osmolality and/or sodium concentration. The present studies were conducted to determine if volume expansion associated with increased plasma osmolality would further augment ANF secretion in the ovine fetus. In response to successive 30-min intravenous infusions of 3% saline at 0.5 and 1.0 ml/kg/min fetal plasma ANF significantly increased from a basal level of 98 +/- 31 pg/ml to a peak of 439 +/- 42 pg/ml (p less than 0.05). During a 30-min postinfusion recovery period, fetal plasma ANF significantly decreased from peak values to 224 +/- 10 pg/ml (p less than 0.05), although remaining above basal levels. Fetal plasma osmolality significantly increased from 300 +/- 2 mosmol to 325 +/- 3 mosmol (p less than 0.05) whereas fetal plasma arginine vasopressin increased from 1.9 +/- 0.4 to 10.9 +/- 7.0 pg/ml (p less than 0.05) at the conclusion of the 3% saline infusion. During the saline infusion a significant increase in fetal heart rate and decrease in fetal hematocrit were noted. Fetal blood pressure and maternal plasma ANF and arginine vasopressin concentrations remained unchanged. Despite the potential stimulatory effects of hyperosmolality, increased plasma arginine vasopressin, and intravascular volume expansion, the increase in fetal plasma ANF in the present study did not exceed that induced by isotonic saline alone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2974941 TI - [Immunologic data on the pathogenesis of hemolytic disease in the newborn]. PMID- 2974942 TI - [Results of a study of morbidity in children from the moment of birth to entry into school]. PMID- 2974943 TI - [The joint work experience of the department of pediatrics of a medical institute and of the public health section of a territorial executive committee and of a branch trade union in promoting the health of children who are frequently ill]. PMID- 2974944 TI - [Provision of optimal microclimate parameters in pediatric pulmonology hospitals and their effect on the course of acute pneumonia in young infants]. PMID- 2974945 TI - Use of the personal orientation inventory with disabled athletes. AB - Athletes who are blind or have impairments in hand-eye coordination that prevent writing are unable to complete psychological inventories in the standardized manner, i.e., read silently and answered independently and anonymously. Two studies were conducted on the oral administration of the Personal Orientation Inventory as a measure of self-actualization of disabled athletes. Reliabilities were examined across modalities (oral vs written) by administering the inventory both ways to 25 high school and college athletes, M age = 21.6 yr. Test-retest reliability for oral administration was affirmed in a study of 15 blind elite male athletes and 15 cerebral palsied elite male and female athletes. It was concluded that the inventory, administered orally, may be appropriately used with disabled athletes. PMID- 2974946 TI - Mouse lipocortin I cDNA. PMID- 2974947 TI - Nucleotide sequence of three cDNAs for the human high affinity Fc receptor (FcRI). PMID- 2974948 TI - Lambda gt22S, a phage expression vector for the directional cloning of cDNA by the use of a single restriction enzyme SfiI. PMID- 2974949 TI - Postoperative backache. PMID- 2974950 TI - Dress code. Interview by Joanna Trevelyan. PMID- 2974951 TI - [Evaluation of the cardiovascular system in children after total surgical correction of tetralogy of Fallot based on roentgenograms of the chest]. PMID- 2974952 TI - [Effect of cinnarizine on subpopulations of thymus-dependent lymphocytes in the elderly]. PMID- 2974953 TI - Arthrogryposis multiplex congenita, Pena-Shokeir phenotype, with gastroschisis and agenesis of the leg. AB - A young mother had a stillborn infant at 33 weeks' gestation, the pregnancy complicated by polyhydramnios. The parents were unrelated, healthy, and both had a normal karyotype. The infant had multiple malformations such as ankylosis, facial anomalies, and pulmonary hypoplasia. A severe gastroschisis and agenesis of the right leg were also present. The neuropathologic findings were those of marked atrophy of anterior horn motor cells in the spinal cord and neurogenic muscle atrophy. This is a case of Pena-Shokeir syndrome with two additional features: gastroschisis and agenesis of the right leg. This syndrome represents a lethal form of arthrogryposis multiplex congenita, and the essential neuropathologic findings are marked reduction in the number of spinal motor cells and neurogenic muscle atrophy. PMID- 2974955 TI - First trimester diagnosis of Gaucher disease in a fetus with trisomy 21. AB - A 38-year-old lady, who had a previous infant with type 2 Gaucher disease, underwent prenatal diagnosis by chorionic villus sampling at 9 weeks' gestation. Results on the fresh villus revealed a 47,XY,+21 karyotype and a marked deficiency (2 per cent of control) of beta-glucosidase activity. Following termination, villus material was cultured which initially revealed only a partial enzyme deficiency and a normal female karyotype, i.e., maternal cells. A subsequent culture contained 47,XY,+21 cells which were deficient in beta glucosidase activity, thus confirming the diagnosis. The results in this interesting case illustrate the potential dangers of maternal cell contamination in cultured villus cells. PMID- 2974954 TI - Maternal serum thyroid antibodies in early pregnancy and fetal Down's syndrome. AB - Thyroid antibodies were measured in mid-trimester antenatal serum samples from 77 pregnancies affected by fetal Down's syndrome and 385 unaffected control pregnancies. Using a haemagglutination technique, thyroglobulin antibodies were detected in 5.2 per cent of cases (4) and 2.9 per cent of controls (11), and thyroid microsomal antibodies were detected in 22 per cent (17) and 15 per cent (59), respectively. Using an enzyme-linked immunosorbent assay (ELISA) for thyroglobulin antibodies and a cut-off level of 50 KIU/l, positive results were found in 25 per cent of cases (19) and 22 per cent of controls (84). Using an ELISA for thyroid microsomal antibodies and the same cut-off level, the proportions were 52 per cent (40) and 39 per cent (149), respectively. While not statistically significant, the differences were consistent with the previously reported increased levels of thyroid antibody found in non-pregnant women who had had pregnancies associated with Down's syndrome. PMID- 2974956 TI - Cardiac defects in chromosomally abnormal human embryos of 10-14 weeks' gestation. AB - Cardiac defects were studied in five chromosomally abnormal embryos of 10-14 weeks' gestation by free-hand microdissection of hearts measuring 2.5-6 mm in diameter. The type of cardiac malformation alone or in association with other anomalies helped to confirm the chromosome diagnosis established prenatally by chorionic villus sampling or after spontaneous abortion. It was suggestive of a chromosomal disorder in one case in which cytogenetic investigation had failed. PMID- 2974958 TI - [Role of the family physician in the prevention policy of colonic and rectal neoplasms]. PMID- 2974957 TI - [Physiopathogeny of sclerodermal cardiomyopathy]. PMID- 2974960 TI - [Post-pneumonectomy bronchial fistula. Treatment by resection of the carina]. AB - A technique for the surgical treatment of bronchial fistulae developed after pneumonectomy is presented. This technique, which resembles that of Abruzzini, consists of resection of the carina followed by tracheo-bronchial suture in a healthy area away from any purulent focus of infection. It is easy to perform and facilitates the treatment of thoracic empyema which is no longer maintained by the fistula. PMID- 2974961 TI - [Acute respiratory insufficiency following laryngeal actinomycosis]. PMID- 2974959 TI - [40 years' development of a case of pseudo-achondroplasia. Role of mechanical and cellular factors in joint destructions]. AB - A man with pseudoachondroplasia was examined on several occasions between the ages of 13 and 50 years. The patient presented with all the signs typical of the disease: severe rhizomelic dwarfism discovered during the second year of life, relatively normal height of trunk, short and massive hands and feet, waddling gait, gross epiphyseal [corrected] gland alterations and shallow vertebral bodies. A review of the literature yielded some 70 cases published since 1959. The main point in the course of the disease was the absence of important changes in radiological abnormalities between the ages of 20 and 50 years. Furthermore, in spite of pronounced epiphyseal lesions [corrected] the patient did not develop severe osteo-arthritis. These findings suggest that beside mechanical factors cellular factors intervene in the development of osteo-arthritis [corrected] PMID- 2974962 TI - [Human immunodeficiency virus type 2 infection discovered by dementia in a French heterosexual male]. PMID- 2974963 TI - [Induction of tolerance to sulfadiazine in a patient with acquired immunodeficiency syndrome]. PMID- 2974964 TI - [Obstructive apnea in the infant. An unusual disclosure of mucoviscidosis]. PMID- 2974965 TI - [At home thrombolysis. 100 cases]. PMID- 2974966 TI - [A new nephropathy: immunoglobulin light chain disease]. PMID- 2974967 TI - [Meta-analysis in clinical research]. PMID- 2974968 TI - [Pharmacokinetics of cefotaxime in cirrhotic patients with or without ascites]. AB - Pharmacokinetic values of cefotaxime were measured in 12 cirrhotic male patients (6 without ascites and 6 with ascites) after intravenous injection of a single 2 gram dose of the antibiotic. In patients without ascites elimination of the drug was about the same as in normal subjects or control patients, although clearance was increased. In patients with ascites, the drug elimination half-life was significantly more prolonged (7.5 +/- 3.9 h versus 1.3 +/- 0.4 h, P less than 0.01) and the drug clearance was significantly lower (193.6 +/- 92.4 ml/min versus 475.8 +/- 152.2 ml/min, P less than 0.01) than in the other group. The accumulation of cefotaxime in these patients produced concentration in the ascites fluid that were above the critical therapeutic values for about 20 hours. PMID- 2974969 TI - [Insufficiency fractures of the sacrum. 5 cases]. AB - Five cases of insufficiency fracture of the sacrum in women aged over 60 are reported. The elements of diagnosis and causative factors are reviewed. In all cases the diagnosis was confirmed by technetium bone scintigraphy and computerized tomography of the sacrum. Bone tissue evaluation in search of a bone wasting disease, the principal cause of this type of fracture, was conducted in all 5 patients, using photon absorptiometry and/or double energy computerized tomography and histomorphometry. These examinations provided detailed information on disorders of mineralization in the genesis of these insufficiency fractures. PMID- 2974970 TI - [Development of insulin reserves in the first 18 months in the insulin-dependent diabetic with or without remission of insulin-dependence]. AB - It seems rational to consider that residual insulin secretion is one of the factors which determine the short-term course of inaugural type I diabetes. But what about the mid-term course? We evaluated prospectively the insulin reserve (fasting and post-prandial C peptide) in 52 patients throughout the subsequent development of the disease. The patients (36 men, 16 women, mean age 35 years), who presented with ketonuria and weight loss, received a 10-day course of intensive insulin therapy, after which a remission of insulin dependence was observed in 40 of them (77 per cent). These 40 patients differed from those who had no such remission in that they were heavier and had a better initial insulin secretion. There was no significant difference between the two groups with regards to immunogenetic markers (presence of anti-islet antibodies 28/35 vs 8/12, DR3 and/or DR4 tissue group 27/37 vs 8/10). Following intensive insulin therapy, the C peptide value was consistently increased. At 6, 12 and 18 months the insulin secretion in patients of the remission group remained stable and always higher than that of patients who did not have a remission and whose insulin secretion collapsed at 18 months. Another characteristic of the remission group was that C peptide secretion could be stimulated by meals throughout the follow-up period (post-prandial C peptide at 18 months: 0.63 nmol/l). It is concluded that residual insulin secretion is one of the most effective predictive factors of remission when type I diabetes is first diagnosed and remains stable for the first 18 months of the disease in patients who show a remission. PMID- 2974971 TI - [Ankylosing spondylitis with type AA amyloidosis. 6 cases]. AB - The authors have studied 6 cases of systemic AA amyloidosis associated with ankylosing spondylitis. Renal failure occurred in all patients a mean of 19 years after the clinical onset of the rheumatic disease. Three patients progressed rapidly (between 3 months and 3 years) to end-stage renal failure. Such an outcome did not depend upon early onset of the renal impairment, degree of inflammation or treatment with colchicine. All patients were alive 2 to 10 years later, and this confirms a better prognosis than with AL amyloidosis. The utility of combining Wright's permaganate reaction with immunological methods to characterize the amyloid deposits was also confirmed. It is concluded that amyloidosis is a rare complication of ankylosing spondylitis and probably depends on a genetic predisposition. The possibility of amyloidosis should be kept in mind when proteinuria or renal failure appear in the course of ankylosing spondylitis. PMID- 2974972 TI - [Determination of the real value over 1 year of the cost of a myocardial infarction. Study in 78 thrombolysed or non-thrombolysed patients]. PMID- 2974973 TI - [Lactic acidosis caused by vitamin B1 deficiency in exclusive parenteral feeding. A case of 1 child]. PMID- 2974974 TI - [Role of hemodialysis and plasma exchange in the removal of methotrexate]. PMID- 2974975 TI - [Vaccination against hepatitis B of children with of homozygous beta thalassemia]. PMID- 2974976 TI - [Autografts from blood cells]. PMID- 2974977 TI - [Parasitic arthritis and rheumatism, reactive arthritis]. PMID- 2974978 TI - [Human calcitonin in neoplastic hypercalcemia. Results of a prospective randomized trial]. AB - Forty patients with hypercalcaemia (2.75 to 3.5 mmoles/l) due to bone metastases or to a neoplastic syndrome were treated with synthetic human calcitonin after previous hyperhydration. On entering the study, the patients were allocated at random to either an 0.5 mg dose or a 1 mg dose of the compound administered 6 hourly by the intravenous route. Calcemia was measured every 6 hours: if it became normal, treatment was discontinued 12 hours later, but if after 3 injections the blood calcium level had not been reduced by more than 0.4 mmoles/l, incremental doses of calcitonin were given. Synthetic human calcitonin reduced calcemia by more than 0.4 mmoles/l in 22 patients (group 1) and brought it down to normal value in 18 of these 8 hours on average after the first injection. A new rise in calcemia was observed in 12/18 cases 44 hours on average after treatment was discontinued. Reduction of calcemia by less than 0.4 mmoles/l was observed in 11 patients (group 2). Treatment was ineffective in 7 patients (group 3). There were no significant differences between the 3 groups in mean values and variances of the initial blood calcium levels. Whatever the initial dose of calcitonin, the therapeutic effect was obtained within the first 24 hours of treatment. In patients with incomplete results increasing the dosage did not result in a further reduction of calcemia. No hypocalcaemia was observed. This study confirms that synthetic human calcitonin has a normalizing effect on calcemia. This effect is not dose-dependent, and blood calcium levels rise again after treatment is discontinued. PMID- 2974979 TI - [Wegener's granulomatosis in elderly subjects. 37 cases]. AB - The clinical, biochemical and prognostic characteristics of 37 patients with Wegener's granulomatosis were studied in relation to age. Eleven patients were older than 60 years when the first sign of the disease appeared. Except for the fact that the renal lesions seemed to be more severe in these patients, there was no significant clinical or biochemical difference between them and patients under 60. Five patients over 60 years of age died during the first 2 years of Wegener's granulomatosis. Death was due to infection in 3 cases, to acute renal failure as complication of intra-alveolar haemorrhage in 1 case and to metastatic cancer in 1 case. Age-related comparisons of survival rates showed that the probability of surviving was lower in elderly subjects: 78 per cent at 1 year and 39 per cent at 3 and 5 years, as opposed to 96 per cent at 1 and 3 years and 84 per cent at 5 years in younger subjects (P less than 0.01). This poor prognosis is probably due to the renal lesions which constitute an important cause of death in the published series. Rapidly progressive glomerulonephritis being amenable to an early and intensive treatment, elderly patients should be treated as energetically as younger patients. PMID- 2974980 TI - [Bronchiolitis obliterans with cryptogenetic-like organizing pneumonia. Demonstration of gastro-esophageal reflux in 5 cases]. AB - Gastro-oesophageal reflux was demonstrated in 5 cases of bronchiolitis obliterans with apparently cryptogenetic organizing pneumonia. In addition, one patient had hiatus hernia and another, oesophageal diverticulum. In 4 patients, after failure of prolonged antibiotic therapy, medical or surgical treatment of the gastro oesophageal reflux resulted in regression of the clinical and radiological signs of pulmonary lesions. The cure thus obtained persisted throughout a follow-up period of 2 months to 8 years. Gastro-oesophageal reflux therefore could be one of the causes of apparently cryptogenetic bronchiolitis obliterans with organizing pneumonia, and all patients with this respiratory disease should be investigated for gastro-oesophageal reflux. PMID- 2974981 TI - [Cardiac involvement in Behcet's disease. 12 cases]. AB - Fifteen cardiac manifestations observed in 12 of a series of 196 patients with Behcet's disease are reported. In this retrospective study conducted in an internal medicine department, 5 cases of pericarditis and 4 cases of myocardial infarction were encountered. The other cardiac manifestations were observed only once. They included ventricular aneurysm, endomyocardial fibrosis of the right heart, aortic insufficiency, mitral valve insufficiency, mitral valve prolapse, and right heart failure consecutive to pulmonary arterial hypertension. There was no relationship between the severity of cardiac lesions and that of the extracardiac manifestations of the disease. Comparison of these data with those found in the literature showed that pericarditis is the most frequent pathology, but it usually regresses rapidly. Lesions of the coronary arteries, with or without myocardial infarction, consist of stenosis, occlusion or pseudoaneurysm requiring surgical treatment. Myocardial lesions (with the possibility of pseudoaneurysm) and endomyocardial right heart fibrosis are exceptional but fairly characteristic of Behcet's disease which they should suggest. Hughes Stovin's syndrome may be complicated by pulmonary arterial hypertension, although death is generally caused by massive haemoptysis. PMID- 2974983 TI - [Hyponatremia of psychotic potomanic subjects. Potomanic patients should no longer be sent to intensive care units]. PMID- 2974982 TI - [Rosacea]. AB - Rosacea should no longer be considered a follicular skin disease. It is a vascular disease of the face characterized by a significant evolution towards local complications such as telangiectasias, papular and aseptic pustular lesions, lupoid granulomas, chronic facial oedema and seboglandular hyperplasia. The basic abnormality seems to be a microcirculatory disturbance of the function of the facial angular veins directly involved in the brain-cooling vascular mechanism. The first clinic hallmark of this dysfunction is the occurrence of flushing, which may be spontaneous or induced by alcohol, intake of hot food, emotional stress and sudden variations in temperature. Tetracycline, metronidazole and isotretinoin are very useful for therapy but they only influence the cutaneous and ocular complications and do not act upon the basic vascular trouble. Current therapeutic research is directed towards drugs having an alpha-sympathomimetic activity and inhibiting the endogenous opioid mediators of flushing such as naloxone or clonidine. PMID- 2974984 TI - [Malignant transformation of an adenomyoma of the cardia]. PMID- 2974985 TI - [Type IIa dyslipoproteinemia. Effects of simvastatin and bezafibrate on atherogenic and non-atherogenic lipoprotein particles]. PMID- 2974986 TI - [Functions of a district phthisiologist in ambulatory practice]. PMID- 2974987 TI - [Solar activity and epidemiological indicators of tuberculosis]. PMID- 2974989 TI - [Objectives of anti-tuberculosis propaganda]. PMID- 2974988 TI - [Incidence and nature of adverse reactions to antitubercular drugs in children with primary intrathoracic tuberculosis]. PMID- 2974990 TI - Inhibition of the Ca pump ATPase of human red blood cell membranes by dithio compounds. PMID- 2974991 TI - Atrial natriuretic factor levels in diabetic rats. PMID- 2974993 TI - Failure of select diuretics, except furosemide, to attenuate action of atrial natriuretic peptide. PMID- 2974992 TI - Regional specificity of beta-endorphin metabolism in brain slices of the rat. PMID- 2974994 TI - Photodynamic therapy of malignant tumors. PMID- 2974995 TI - Effects of D1 and D2 antagonists on basal and apomorphine decreased body temperature in mice and rats. AB - In these experiments representative selective antagonists at D1 (SCH 23390) and D2 (haloperidol) receptors were studied for their effects on basal and apomorphine decreased body temperature in mice and rats. In mice, SCH 23390 (up to 3 mg/kg SC) neither affected basal body temperature nor blocked apomorphine induced hypothermia (AIH). On the other hand, haloperidol alone was hypothermic and paradoxically also blocked AIH in mice. In rats, SCH 23390 alone produced hyperthermia; the mechanism by which this occurred is not known. SCH 23390 also blocked AIH in rats. However, the inhibition of AIH only occurred at doses of SCH 23390 that were themselves hyperthermic. Haloperidol did not alter basal body temperature but did block AIH in rats. These data suggest that apomorphine induced body temperature changes are D2 mediated. PMID- 2974996 TI - The D2 dopamine receptor agonist LY171555 induces catalepsy in the mouse. AB - The dopamine agonist LY171555 (quinpirole), a specific D2 receptor agonist, induces catalepsy in mice at doses ranging from 0.3 to 10 mg/kg. The effects of an intermediate dose of this compound (1 mg/kg SC) were antagonized by 25 mg/kg of the selective D2 antagonist (-)-sulpiride (IP) injected 20 min before LY171555. SCH 23390, a selective D1 antagonist, administered (0.3 mg/kg IP) 20 min before LY171555 (1 mg/kg) enhanced the cataleptic effects of this compound. Finally, when the D1 dopamine receptor agonist SKF 38393 (20 mg/kg SC) was administered immediately beforehand, the cataleptic effects of 1 mg/kg of LY171555 were markedly reduced. These results suggest that there is a functional interaction between D1 and D2 dopamine receptors in the modulation of catalepsy. PMID- 2974997 TI - Evidence that a GABAergic mechanism influences the development of obesity in obese Zucker rats. PMID- 2974998 TI - Comparison of the gastric antisecretory effects of ramixotidine dihydrochloride (CM 57755), a new H2 receptor antagonist, and cimetidine in dogs. AB - This paper compares the effects of ramixotidine dihydrochloride (CM 57755) with those of cimetidine on gastric acid secretion and gastrin release in conscious dogs chronically fitted with Heidenhain pouches and/or gastric fistulae. At equimolar doses, intravenous (i.v.) or intragastric (i.g.) CM 57755 caused similar inhibition of dimaprit- or pentagastrin-induced secretion than cimetidine. Acid secretion stimulated by a meat meal was significantly reduced by both CM 57755 and cimetidine. Neither CM 57755 (4.5 and 9 mumol/kg) nor cimetidine (4 mumol/kg) modified gastrin release, while cimetidine (8 mumol/kg) significantly increased it. Judging from these results, while CM 57755 appears to be an inhibitor of gastric acid secretion induced by different stimulants in dogs with potency comparable to cimetidine. The increase in plasma gastrin levels seen after cimetidine but not after CM 57755 suggests that cimetidine releases gastrin by a mechanism independent of H2 receptor antagonism. PMID- 2974999 TI - [Synthesis of N-alpha-(arylsulfonyl)-4-amidino-phenylalanyl-prolines and N-alpha (arylsulfonylglycyl)-4-amidino-phenylalanyl-prolines and testing of them as inhibitors of serine proteinases. 33. Synthetic inhibitors of serine proteinases]. AB - The headline compounds were prepared by the reaction of N alpha-(arylsulfonyl)- and N alpha-(arylsulfonylglycyl)-4-cyanophenylalanines to the L- and D-proline derivatives. These compounds were transformed in a usual manner in the amidino compounds. The substitution of the pyrrolidine residue by L-proline in this type of inhibitors decreased the activity against serine proteases, especially against thrombin. In the case of the D-proline derivatives the decrease of the inhibitor activity against thrombin was very high. PMID- 2975001 TI - Anti-metrazol action of R 57720 during postnatal development in rats. AB - The effects of an experimental antiepileptic drug R 57720 (Janssen Pharmaceutica) on metrazol-induced seizures were studied in rats aged 7, 12, 18, 25 and 90 days. R 57720 exhibited a marked dose-dependent anticonvulsant effect against major, generalized tonic-clonic seizures in all the age groups studied. Similar action on minimal metrazol seizures (mMS) was present in rats aged 18 days and more. In the two youngest groups where metrazol did not induce mMS under control conditions, the combination of the 5- and 10-mg/kg doses of R 57720 and metrazol led to the appearance of mMS, whereas mMS failed to appear after the highest dose of R 57720 used (20 mg/kg). PMID- 2975000 TI - Plasma xanthine oxidase activity correlates with the resistance to severe hypoxia in different species. AB - A rapid polarographic method of measuring the O-form of xanthine oxidase (XOD) is described. Activities of this enzyme and oxypurine concentrations were measured in the plasma and cerebrospinal fluid (CSF) of 1, 12, 25 and 60-day-old rats. In the CSF, the highest oxypurine concentration was found in the group of 12-day-old rats (34.07 +/- 12.37 mumol.l-1), and it remained nearly at the same level in groups of older animals. Oxypurines in the plasma, on the contrary, are polarographically measurable on the first day of life; in groups of older rats their concentration is below the sensitivity of the method (5 mumol.l-1). This was explained by the development of XOD activity in the plasma, which increased from 8.17 +/- 2.80 to 99.46 +/- 13.85 nkat.l-1 during the first 60 days of postnatal life. The higher the normal plasma XOD activity, the shorter was the survival time of the species during interrupted hypobaric hypoxia. Adult guinea pigs and hamsters have no measurable XOD activity in the plasma; adult rats and mice have 99.46 +/- 13.85 and 259.69 +/- 58.23 nkat.l-1, respectively. The survival time of these animals at 10,500 m was measured following exposure to an altitude of 8,000 m for 30 min and 15 minute normoxia. Guinea-pigs survived 100.8 +/- 13.84 min, hamsters 54.25 +/- 11.33, rats 25.2 +/- 5.37 and mice 3.33 +/- 1.00 min. PMID- 2975002 TI - Influence of cinromide on metrazol--induced seizures during ontogenesis in rats. AB - The effects of cinromide on two types of metrazol-induced seizures were studied in 236 male rats aged 7, 12, 18, 25 and 90 days. Cinromide was administered intraperitoneally in a dose of 25 or 50 mg.kg-1 30 min before a subcutaneous injection of metrazol. Major, i.e. generalized tonic-clonic seizures could be elicited by metrazol in all age groups. Cinromide was efficient against this type of seizures at all developmental stages, but its action in 7- and especially in 12-day-old rat pups was less marked than in older animals. Minimal metrazol seizures (predominantly clonic) could be reliably elicited since the age of 18 days. Cinromide was also able to suppress this type of seizures, but in adult rats the dose of 75 mg.kg-1 had to be administered because even the 50 mg.kg-1 dose was not sufficient. Quantitative changes of the antimetrazol action of cinromide were demonstrated during maturation in rats. PMID- 2975004 TI - The diuretic effect of metipamide and its relationship to body weights in rats. AB - Metipamide [M], a new Czechosclovak diuretic with a hypotensive effect, was administered in a dose of 20 mg/kg (about 500-fold the therapeutic dose) I. for three weeks to rats of both sexes kept under normal conditions in groups of five, and II, for eight days to single male rats in metabolic cages. The animals' body weight and food and water consumption were studied and in the second series their daily faeces, urine and urinary sodium and potassium excretion were measured. Rats kept in individual cages were also given indapamide (I), the first diuretic with a separate hypotensive effect used in other countries, in a dose of 20 mg/kg. The experimental animals' body weight was significantly lower than that of controls with the same food consumption and their water intake and urine flow were much higher, especially after M. Sodium (and to a lesser extent potassium) excretion was raised at the outset of administration of both the test substances and again after the 5th to 8th dose, but only after M. After three weeks' administration of M. SNa, SK and S(osm) values were within normal limits. but after eight days the serum electrolytes and the osmolality of the serum were markedly reduced. After I. these values were normal. We conclude that the strong diuretic effect of M is not the only cause of lower body weight in rats. PMID- 2975003 TI - Pyritinol and the enzymes of gamma-aminobutyric acid (GABA) synthesis and degradation. AB - Pyritinol, a vitamin B6 derivative considered to have an activating effect on brain inhibited glutamate decarboxylase in concentrations of 0.05-1.0 mmol/l. This effect was not dependent on the pyridoxal-5'-phosphate concentration. An increase in the glutamate level reduced the inhibitory effect of pyritinol, but inhibition was not competitive. It is supposed that this modification of inhibition of glutamate decarboxylase by the substrate concentration might be associated with the presence of two glutamate decarboxylases with different affinities for the substrate. The inhibitory effect of pyritinol was dependent on integrity of the disulphide bond in the pyritinol molecule. Inhibition of glutamate decarboxylase increased in correlation to time--possibly in association with progressive oxidation of the SH-groups of the enzyme. Pyritinol did not influence GABA transaminase activity, but lessened the oxidation of GABA to carbon dioxide. It is assumed that succinic semialdehyde dehydrogenase activity was inhibited. PMID- 2975005 TI - Selenium metabolism in rats after administration of toxic doses of selenite. AB - We studied organ concentration, excretion and excreted forms of selenium in young and adult rats after a single s.c. injection of a sublethal dose of 75Se selenite. In the young about a 10-fold higher concentration of 75Se in blood, liver, kidneys and In the young, about a 10-fold higher concentration of 75Se in blood, liver, kidneys and heart was found at all the experimental intervals studied (1-7 days). The highest 75Se concentration in the young was in the liver while in the adults it was found in the kidneys. The spectrum of radioselenium metabolites in the urine was the same in both groups. However, the main product excreted by young rats was 75Se-glutathione selenotrisulphide and an unidentified neutral substance while it was the trimethylselenonium ion in the adults. Ontogenetic differences in selenium metabolism could be one of the factors underlying the differences in the response of the young and the adult rats to toxic doses of selenite. PMID- 2975006 TI - Effect of polyenoic phospholipid therapy on lecithin cholesterol acyltransferase activity in the human serum. AB - The effect of polyenoic phospholipids on the concentration of serum lipids and the activity of lecithin cholesterol acyltransferase (LCAT, E.C. 2.3.1.43) was investigated in 18 patients with chronic glomerulonephritis accompanied by hyperlipaemia and reduced rate of cholesterol esterification in the plasma. The effects of therapy were evaluated immediately after a 2-month period of treatment and again after a 3-month drug free interval following termination of the therapy. An immediate effect of the treatment was reflected in a significant increase in the fractional esterification rate (FER % .h-1) and a marked reduction of the concentration of triglycerides (TG). Discontinuation of the drug resulted in the return of TG and FER values to the initial levels and in a rise of total (TCH) and unesterified cholesterol (UCH), HDL-cholesterol (HDL-TCH) and the molar esterification rate (MER mumol.1-1.h-1). The activity of LCAT estimated by radioassay in common and endogenous substrates varied in parallel. PMID- 2975007 TI - The use of an intestinal thermode for studying thermoregulation of the golden hamster. AB - Intestinal cooling induces a normal metabolic response compensating heat loss in the euthermic golden hamster. The hypothalamic and subcutaneous temperatures change unpredictably and the threshold hypothalamic temperature for the induction of cold thermogenesis, similarly as the sensitivity of the regulator related to changes in hypothalamic temperature vary considerably, however. It seems that the thermal input from the hypothalamus does not contribute significantly to the control of cold thermogenesis in euthermic golden hamsters during intestinal cooling. The use of an intestinal thermode itself is not suitable for quantitative studies of thermoregulation in the golden hamster. PMID- 2975008 TI - An analysis of the edge damage of membranes studied in vitro. AB - A mathematical formula was developed for estimating the effect of membrane edge damage on the membrane permeability measured in vitro. It was assumed that in the chambers compressed against each other with identical force, the membrane edge damage depends on their radius and is characteristic for different tissues. Transport of rubidium and albumin across the human amnion and the rabbit mesentery was measured in two chambers with different active areas and this made it possible to estimate the extent of membrane edge damage. PMID- 2975009 TI - Studies of tissue electrical admittance: relation to tissue sodium for different zones of rabbit kidney. AB - Tissue electrical admittance (reciprocal impedance) and Na+ concentration were determined in slices of rabbit renal cortex, outer medulla, inner medulla and the papilla. In each zone admittance was highly and significantly correlated to tissue Na+ (r = 0.71 to 0.91, p less than 0.001). The cortex admittance proved a relatively insensitive index of tissue electrolyte concentration. The highest sensitivity was observed for the outer medulla: values for the inner medulla and papilla were slightly lower. The data confirm the usefulness of admittance measurement for dynamic assessment of the cortico-papillary electrolyte gradient but show that the values measured in the outer medulla cannot be directly compared with those for the inner medulla and the papilla. PMID- 2975010 TI - Consequences of chronic phenobarbital blockade of spontaneous motility in chick embryos for the development of central motor output activity. AB - The consequences of the chronic continuous administration of Na+-phenobarbital in an average dose of 9.8 mg/kg e.w./24 h were studied in chick embryos. Administration was always started on the 4th day of incubation and lasted 4-12 days. Spontaneous motility was tested in 13- and 17-day-old embryos. The continuous, chronic administration of phenobarbital produced the following significant changes in the embryos' spontaneous motor activity and in the reactivity of their central motor output: 1) In 17-day-old embryos it reduced spontaneous motility in direct correlation to the time of administration. In 13 day-old embryos the effect was not yet significant. 2) It significantly inhibited strychnine, bicuculline and metrazol activation of motor output. 3) It raised the sensitivity to the acute administration of oxazepam (by 18-29.4%), but weakened the inhibitory effect of gamma-aminobutyric acid (by 17.6-33.2%). From the results of these observations it is deduced that spontaneous central motor output activity is not just a developmental epiphenomenon, but that it is an important prerequisite for normal development of the embryonic CNS. PMID- 2975011 TI - Atypical laterality and retardation. PMID- 2975013 TI - Health Dept strategy to cut workers' compensation costs an unwarranted invasion of privacy. PMID- 2975012 TI - Differential effects of the selective D2-antagonist raclopride in the nucleus accumbens of the rat on spontaneous and d-amphetamine-induced activity. AB - The effect of the D2-antagonist raclopride was investigated in two test situations, which are presumed to involve dopamine (DA) transmission within the nucleus accumbens of the rat. Local injection of d-amphetamine sulphate (10 micrograms/0.5 microliter) produced a marked increase in motor activity, measured as motility, locomotion, and rearing, which was dose- and time-dependently antagonised by local injection of raclopride (0.05-5.0 micrograms/0.5 microliter). After an initial decrease, at low doses (0.05-0.25 microgram/0.5 microliter) an apparent enhancement of the d-amphetamine-induced motor activity appeared, which was most clearly seen with rearing. These lower doses, however, did not induce any clear changes in the exploratory activity in a novel environment (i.e., the second test situation). Only the higher doses used (1.0 5.0 micrograms/0.5 microliter) decreased exploratory activity during the first 5 10 min, also measured as motility, locomotion, and rearing. These data are discussed with respect to the role of D2-receptors within the nucleus accumbens of rats in the motor activity induced by a novel environment and d-amphetamine. Overall, the data underline previous notions that raclopride is a potent antagonist of DA-mediated behaviour. PMID- 2975014 TI - [Percutaneous transluminal laser angioplasty. Initial clinical experience in 9 patients]. AB - In this paper the authors report on a new technique--percutaneous transluminal laser angioplasty (PLR) which was performed on 9 patients with iliac and/or femoro-popliteal artery occlusion. All patients were males (mean age: 64.5 years) and had arterial occlusion (mean length: cm 12.5). PLR was performed with an argon laser (max power: 16 Watts). In 8 out of 9 patients (88.8%) a complete recanalization was obtained of the occluded arterial tract, without complications. In one case only (12.2%) there was an arterial wall perforation with unsatisfactory results. In our opinion, PLR has proven a simple methodology, which can be performed on any patient due to the very low incidence of severe complications and distal embolism. Moreover, PLR has quite low costs, and does not prevent eventual surgical/percutaneous interventions. PMID- 2975015 TI - [Spontaneous hematoma during anticoagulant therapy. Usefulness of CT]. PMID- 2975016 TI - [Conductive hypoacusis in minor malformations of the middle ear]. PMID- 2975017 TI - Flow cytometric evaluation of cell-cycle phases in laryngeal carcinomas. PMID- 2975019 TI - [Clinical and diagnostic aspects in intrapetrosal aneurysms of the internal carotid artery]. PMID- 2975018 TI - [Evoked potentials and the oculofacial reflex in otoneurological pathology]. PMID- 2975020 TI - [Long-term evolution of chronic suppurative otomastoiditis]. PMID- 2975022 TI - [A case of hemangioma of the nasal pyramid treated by cryotherapy with liquid nitrogen in an infant]. PMID- 2975023 TI - [Primary atypical syphilis (bifocal, tonsillar and perianal)]. PMID- 2975021 TI - [Our procedures in treating recent injuries of the middle ear]. PMID- 2975024 TI - [Socio-educational integration of the handicapped with hearing deficiencies]. PMID- 2975025 TI - [Cultivation of voice intensity in deaf children]. PMID- 2975026 TI - [Acupuncture--an adjuvant therapeutic measure in the rehabilitation of patients with hyperfunctional dysphonia]. PMID- 2975027 TI - [How to evaluate the immediate results of percutaneous transluminal angiography? Should pressure gradient, flow reserve or the minimal transluminal cross sectional area be used?]. PMID- 2975028 TI - [Use of a new double-balloon catheter in percutaneous transatrial mitral valvuloplasty]. PMID- 2975029 TI - [Use of echocardiography in percutaneous mitral and aortic dilatation]. PMID- 2975030 TI - Spontaneous occurrence and experimental induction of leiomyoma of the ventral ligament of the oviduct of the hen. AB - Leiomyoma of the ventral ligament (VL) of the oviduct is a common tumour of the domestic fowl which occurs in specific pathogen free hens and commercial breeding hens at the end of the first laying season, with prevalence varying from 0 to 60 per cent in different lines and breeds. To investigate the aetiological role of steroid sex hormones in this tumour, subcutaneous implants of diethylstilboestrol (DES), progesterone (P) and DES combined with P (DESP), were administered to chickens at three-week intervals, starting from eight weeks old. DESP produced hyperplasia and leiomyoma of the VL but DES alone produced only hyperplasia. P alone induced neither hyperplasia nor leiomyoma. DESP started to induce leiomyomas within five weeks of the onset of treatment and after 32 weeks of treatment 75 per cent of the DESP group had developed leiomyomas, compared with none in the control group. The results suggest that continuous DESP-treatment can induce hyperplasia and leiomyomas independent of ovarian involvement. The leiomyomas possessed receptors for oestradiol and P. A role of P as a cocardinogen is indicated. These experiments provide the first example of synergism between an oestrogen and progesterone in tumorigenesis in the fowl. PMID- 2975031 TI - [Basic principles of the current therapy of depressive states]. PMID- 2975032 TI - [Cerebral abscesses secondary to infectious otorhinolaryngologic diseases]. PMID- 2975033 TI - [Mycoses of the central nervous system. Neurosurgical forms]. PMID- 2975034 TI - [Excessive neutralizing activity against the canine morbilli virus in an area at increased risk for multiple sclerosis]. PMID- 2975035 TI - [Sudden death in cerebral epidermoid tumor (forensic autopsy in 2 cases)]. PMID- 2975037 TI - [Personality in bipolar affective disorders. A study with the aid of a personality inventory]. PMID- 2975036 TI - [Clinico-evolutive and therapeutic characteristics of depressive states (reactions and decompensations) in psychopathic personalities]. PMID- 2975038 TI - Protein content and thyroid hormone release in vitro by differentiated thyroid cancer cells in the presence of estradiol, dehydroepiandrosterone, polypeptidic hormones and pesticides. AB - The reactivity of thyroid cancer cells treated with estradiol (E) and dehydroepiandrosterone (DHA) in the presence of other hormones (TSH, STH, insulin), myopeptide and some pesticides (aminotriazole--ATA, atrazine--ATZ, prometryne--P and ATC) proved to be active in the culture medium. Protein synthesis in sclerosing folliculo-papillary cancer cells under the treatment applied is more markedly inhibited by DHA than by E in the culture medium is compared to the control lots. Tg1 and T3 release into the culture medium under the action of T4 and TSH decreases progressively going from the control to the treated lots, in the presence of E and DHA. STH inhibits Tg1 and T3 secretion in thyroid follicular carcinoma, whether or not E or DHA are present in the culture medium. PMID- 2975039 TI - Regulation of growth and differentiation of pre-activated B lymphocytes. AB - The differential effects of H-2 IAk-specific T helper cells, their soluble factors and Sepharose-coupled anti-mu antibodies on the growth and differentiation of pre-activated B cells were studied. It was found that the prolonged growth of pre-activated B cells required activation signals from major histocompatibility complex (MHC)-restricted T helper cells or Sepharose-coupled anti-mu antibodies. The MHC-restricted T helper cells induced both prolonged growth and differentiation of activated B cells. Anti-mu antibodies together with T helper cell-derived soluble factors induced prolonged growth of activated B cells, but no differentiation into Ig secretion was detected. The inhibition of Ig secretion in anti-mu cultures could be overcome to some extent by either MHC restricted T helper cells or lipopolysaccharide together with soluble factors. It was also observed that T helper cell interactions were needed for long-term in vitro culture of pre-activated B lymphocytes. PMID- 2975041 TI - Production of gamma interferon in Mls disparate interactions. AB - The murine T-cell clone E11 isolated from a primary H-2k histocompatible one-way mixed lymphocyte culture of B10.BR anti C3H/Tif spleen cells was used to study multidirectional interactions in strong stimulatory Mls disparate responses. Several parameters have recently been studied, and proliferation of T cells upon stimulation by macrophages or B cells of Mls-disparate stimulator cells, induction of differentiation of B cells of stimulator strains but inhibition of their macrophage differentiation, and the inhibition by E11 T cells of the production of a mitogenic mediator by Mls-disparate spleen cells have been found. As shown in this paper, these phenomena can be explained by an Mlsa, d, e specific induction of gamma-interferon (IFN-gamma) production in the responder B10.BR (Mlsb) E11 T cells. It is suggested that IFN-gamma, as a regulator of feedback mechanisms plays a critical role in Mls disparate T-cell stimulation. PMID- 2975040 TI - Characterization of human CD4+ T-cell clones that secrete helper factor(s) for B cell proliferation and maturation. AB - Human peripheral blood lymphocytes (PBL) were activated with K46M, a m mitogenic monoclonal antibody against La-reactive T lymphocyte surface structures. The cultures were expanded in the presence of interleukin 2 (IL-2). After 1 month of culture, the activated T cells were cloned by limiting dilution at 0.5 cells/well. Five clones with the CD3+CD4+ phenotype and one clone with the CD3+CD8+ phenotype were obtained. The CD3+CD8+ clone (K99) displayed a strong major histocompatibility complex (MHC)-unrestricted cytolytic activity against MOLT-4 and a weaker reactivity against the bladder tumour cell lines T24 and RT4. The natural killer (NK)-susceptible K562 cells were not lysed. Two of the CD3+CD4+ clones (K91 and K914) showed a helper activity in pokeweed mitogen (PWM) induced IgG production by B cells. These cells differed in the expression of CD45R and CDw29 antigens, as defined by the monoclonal antibodies 2H4 or D10D11 and 4B4. When stimulated with PWM for 48 or 72 h, clone K91 and an additional CD4 positive clone (K913) secreted a factor into the supernatants which helped B cells to produce IgG. The K913 supernatant also induced some IgM production. The supernatant obtained after similar stimulation of K914 cells was inactive. None of these supernatants induced B cells to proliferate when tested together with phorbol myristate acetate (PMA). However, when K91 and K914 cells were activated with phytohaemagglutinin (PHA) for 48 or 72 h, the supernatant from K91 was strongly helpful in B-cell proliferation, whereas the supernatant from K914 cultures was only moderately active. In conclusion, we have established human T helper clones that release different factors supporting either B-cell proliferation or maturation when stimulated with PWM or PHA. PMID- 2975042 TI - Generation of high-rate ovalbumin-specific antibody-secreting cells in cultures of human peripheral blood B cells obtained from non-immunized blood donors. AB - When human peripheral blood B cells are cultured for 6 days with the T cell dependent peptide antigen ovalbumin (OA) in the presence of antigen-presenting cells and helper T cells, plaque-forming cells (PFC) are generated. These OA induced PFC differ from the conventional high-rate antibody-secreting PFC formed after stimulation of B cells with recall antigens (e.g. tetanus toxoid) in that they secrete antibody at a very low level. Previous studies have shown that OA induced PFC are B lymphocytes in an early activation state rather than cells that have differentiated into plasmablasts. The apparent arrest in the maturation of OA-induced PFC in an early activation phase can be overcome by simultaneous stimulation with interleukin 2 (IL-2) and gamma interferon (IFN-gamma). The isotype of the OA-specific antibodies secreted, however, are only of the IgM class, demonstrating that an isotype switch does not occur. PMID- 2975043 TI - Workshop on schizophrenia, PET, and dopamine D2 receptors in the human neostriatum. AB - Recently, two research groups published numbers for D2 receptor sites in the neostriatum of drug-naive schizophrenic patients, obtained in vivo by positron emission tomography (PET). One study appeared to confirm the increase of D2 receptor numbers, while the other study did not. A workshop was convened in Montreal to examine the reasons for the discrepancy between the results obtained by the two groups. The workshop considered patient populations, PET instrumentation and scanning methods, pharmacology, and modeling. The workshop identified differences between the approaches of the two groups that could contribute to the divergent results, including age and chronicity of the patient samples, brain region selected for study, metabolism of the different radioligands in blood and brain, reversibility of binding, PET instrumentation, and complexity of data analysis. The workshop concluded that these initial efforts had made considerable progress in establishing the role of PET in the understanding of the biochemical processes underlying mental illness. In particular, the unique ability to quantify regional neuroreceptor density at different stages in the evolution of the disease has been implemented. At the same time, the work so far and this conference served to identify the main sources contributing to the different findings from the two centers. This information will be important in designing the next phase of the research which will build upon and reconcile these apparent discrepancies. PMID- 2975045 TI - Skin and respiratory symptoms from exposure to alkaline glutaraldehyde in medical services. AB - The prevalence of certain symptoms (eye, skin and airway symptoms, headache, nausea, and fatigue) were studied among hospital workers with and without exposure to glutaraldehyde during cold sterilization work. The exposure to glutaraldehyde and formaldehyde was quantified by hygienic measurements in the breathing zone of the workers. Aldehydes were measured by a specific method, using sorbent tubes with Amberlite XAD-2 coated with 2,4-dinitrophenylhydrazine (2,4-DNF) and analyzed by liquid chromatography. The exposure measurements revealed that the present exposure to glutaraldehyde was intermittent and well below the Swedish occupational exposure limit. In spite of this low exposure, the exposed group exhibited a significantly increased frequency of skin and airway symptoms, as well as headache, in comparison with the unexposed group. A dose response relationship between the frequency of exposure and the number of symptoms could also be demonstrated. No case of contact allergy to glutaraldehyde was found. PMID- 2975044 TI - Fibrinolytic activity in the chronic low-back pain syndrome. PMID- 2975046 TI - [Acute coronary occlusion following transluminal angioplasty: the role of an implanted parietal support via transluminal route]. AB - Acute coronary artery occlusion complicates some 5% of balloon angioplasty procedures and often makes emergency surgical revascularization necessary. Initial experience is reported with emergency implantation of endoluminal stents for acute vessel closure following coronary balloon angioplasty. 10 patients received one stent and 2 patients two stents during the study period. In the study group, implantation was technically successful in all cases: there were no deaths, Q-wave infarctions or cases requiring emergency surgery.--2 patients showed a moderate rise in creatinine phosphokinase in the first 48 hours. During a mean follow-up period of 4 months (range 1-21 months) there have been no deaths and no cases requiring elective coronary bypass surgery. 10 patients underwent control angiography and none have developed significant restenosis within the stented segment. For selected patients, coronary artery stenting appears to be a promising technique as an alternative to emergency bypass surgery after acute vessel occlusion during balloon angioplasty. PMID- 2975047 TI - [Incidence and treatment of coronary restenosis in spite of the implantation of an endoprosthesis]. AB - Between April 1986 and February 1988, 72 endoprosthesis implantations were carried out in the coronary arteries of 65 patients. The indication for implantation was either abrupt closure post angioplasty or prevention of restenosis. The first 3 cases of restenosis within the stented segment are reported. Two were treated by repeat balloon angioplasty, with temporary improvement. The possible mechanisms underlying this particular form of restenosis are discussed. PMID- 2975048 TI - [Long-term clinical development following coronary angioplasty]. AB - 100 patients who had undergone successful coronary angioplasty were followed up for 12, 42 and 84 months. The mean age of the population was 53 +/- 10 years and 89 were males. 86 patients had single vessel coronary artery disease and 14 multi vessel disease. 9 patients had had previous by-pass surgery. Coronary angioplasty was done on 54 left anterior descending, 28 right and 9 left circumflex coronary arteries, and 9 by-pass grafts. Follow-up comprised clinical examination, stress test and personal evaluation by the patients. There was a clear-cut improvement in 66% of patients after 1 year, in 84% at 3 1/2 years and 26% at 8 years, excluding 6 deaths and 9 patients undergoing by-pass surgery. Improvement was moderate in 24% of patients at 1 year, 16% at 3 1/2 years and 58% at 8 years. Stress test was negative in 50% of patients at 1 year, in 64% at 2 years and in 70% at 3 1/2 years. At the latest follow-up (8 years) 6 patients had died, 5 had undergone by-pass surgery and 26 had undergone a further coronary angioplasty; 26% of these 84 patients were symptom free, 58% in class I, 9% in class II, 5% in class III and 2% in class IV (New York Heart Association classification). In conclusion there was a clearcut longterm improvement after coronary angioplasty, thus making this technique the first choice treatment in coronary artery disease in patients with suitable coronary lesions. PMID- 2975049 TI - The plasma membrane in the control of the signaling function of calcium. AB - Eukaryotic plasma membranes contain three Ca-transporting systems: a Ca channel, an ATPase, and an Na/Ca exchanger. The ATPase is high-affinity, low-capacity system, which continuously pumps Ca out of cells. The Na/Ca exchanger is a low affinity, high-capacity system, which is particularly active in excitable cells. The exchanger probably functions in both the Ca efflux and influx directions. The Ca-ATPase is a single polypeptide of Mr 138 kD, which is activated by calmodulin or, in its absence, by acidic phospholipids, polyunsaturated fatty acids, and limited proteolytic treatments. Trypsin produces a number of fragments, some of which (Mr 90, 85, and 81 kD) function as ATPases and transport Ca across reconstituted bilayer membranes. Trypsin proteolysis in the presence of different effectors has permitted us to locate the calmodulin-interacting domain of the enzyme in a 9-kD peripheral sequence that consists of a 4-kD calmodulin-binding subdomain and a subdomain of Mr 5 kD, which is essential for the expression of calmodulin stimulation. The Na/Ca exchanger of plasma membranes has not yet been identified with certainty. On the basis of purification attempts using different approaches, probable Mr's of 82, 70, or 33 kD have been proposed. Antibodies raised against the 33-kD protein partially inhibit the exchange activity of heart sarcolemma vesicles. They interact with the 33-kD protein, but also, under nonreducing conditions, with proteins of Mr approximately 70 and approximately 140 kD. Under reducing conditions, the reactivity with the latter component disappears. It is suggested that the monomeric Mr of the exchanger is 33 kD, and that intermolecular disulfide bridges associate monomers into dimeric and tetrameric forms. PMID- 2975051 TI - [Methods of establishing the general criterion of evaluation of the quality of hospital services for the population in public health institutions]. PMID- 2975050 TI - [Improving the organization of rehabilitation treatment in urban polyclinics]. PMID- 2975052 TI - [Work characteristics of sector nurses in pediatric polyclinics]. PMID- 2975053 TI - [Systematic approach to the study of the health of schoolchildren]. PMID- 2975054 TI - [Various results of a medico-sociological study of the population of a regional center]. PMID- 2975055 TI - [Joint work of the Department of Social Insurance of the Regional Trade Union Committee of the agro-industrial complex and the regional clinical hospital with regard to the health protection of rural workers]. PMID- 2975056 TI - [Vladimir Mikhailovich Mysh]. PMID- 2975057 TI - [Subacute trichloroethylene poisoning with myocardial lesion and atrial fibrillation]. PMID- 2975058 TI - [The role of tests for polyenuria in mass screening of the population]. PMID- 2975060 TI - [Detection of arterial hypertension in workers in the rubber manufacturing industry]. PMID- 2975059 TI - [Ultrasonic examination of the thyroid gland in mass screening of the population]. PMID- 2975061 TI - [Results of 3 years' prevention of ischemic heart disease in men aged 40-59]. PMID- 2975063 TI - Normal magnetic resonance imaging with abnormal discography. AB - In degenerative lumbar spine disease, recent studies have supported the clinical usefulness of discography, especially when used with computed tomography (CT) scanning. The role and capabilities of magnetic resonance imaging (MRI) scanning are currently evolving and being defined. This study reviews a series of patients with prolonged disabling symptoms who had normal MRI scans and abnormal discography. Discograms and discogram-CT scans may at times allow detection of clinically correlative and significant pathology (usually annular disruptions) not suggested by MRI scanning. This fact should be considered in patients with normal MRI scanning and continuing unexplained symptomatology. PMID- 2975062 TI - Disc deterioration in low-back syndromes. A prospective, multi-center CT/discography study. AB - Disc deterioration and pain provocation in different low-back pain syndromes was studied using computed tomography (CT) discography. Data were prospectively collected for 300 patients (816 discs). Patients were classified by their pre discography diagnosis of disc herniation (DH), degenerated disc (DD), lumbar syndrome (LS), lumbar radicular syndrome (LRS), or other. The CT/discograms were classified by discographic pain response, the amount of degeneration and annular disruption. Eighty-two percent of DH patients, 80% of DD, 56% of LS, and 59% of LRS patients had both positive discographic pain provocation and moderate or severe disc deterioration. The study indicates that intradiscal pathology plays a major role in nonspecific low-back pain syndromes. PMID- 2975064 TI - Familial predisposition to discogenic low-back pain. An epidemiologic and immunogenetic study. AB - The first-degree relatives (parents, siblings and children) of 284 patients complaining of discogenic low-back pain (Group I), 114 patients who had undergone surgery for lumbar disc herniation (Group II), and 280 individuals who had never complained of low-back pain (Group III) were surveyed by self-completed questionnaires. Of the families in Group I and Group II, 35 and 37%, respectively, had at least one member with a history of discogenic back pain and 5 and 10%, respectively, had one or two members who had undergone disc surgery. Of the asymptomatic subjects in Group III, only 12% had at least one or more affected relatives and 1% had a relative who had undergone disc surgery; of the affected families, 41% had two or more members with a history of back pain. The proportion of symptomatic relatives in the affected families was higher among sedentary workers and motor vehicle drivers than among heavy or light manual workers. An immunogenetic study comparing the frequencies of HLA-A, B, and C antigens in 39 patients who had undergone lumbar disc surgery with those in 60 asymptomatic individuals showed no significant differences between the two groups. This study indicates that there is a strong familial predisposition to discogenic low-back pain, and suggests that the etiology of degenerative disc disease is related to both genetic factors, not linked to the HLA antigen system, and environmental factors. PMID- 2975065 TI - The outcome of surgery for lumbar disc herniation. II. A 4-17 years' follow-up with emphasis on psychosocial aspects. AB - We propose that the operated patients without complaints, patients with complaints and pensioned patients with complaints differ in their psychosocial situation. Thus, the first group show the best and the third the worst psychosocial condition. A sample of 135 patients after disc surgery was chosen at random from the patient population (n = 435) responding to a questionnaire. The patients were divided into three groups: no pain (n = 51), pain, no pension (n = 40), pain and a disability pension (n = 44). All have attended a psychosocial screening. They filled in the MMPI and were interviewed on some aspects of their social life, and reported on the coping modes they used in order to subdue their low-back complaints before or after the operation. Independent from the preoperative low-back pain condition and the immediate postoperative results, the operated patients showed, several years after the operation, traceable differences in several psychosocial factors according to the socially and personally defined illness career and its stages. The difference is particularly strong between the patients without complaints and the pensioned patients. The patients with complaints, mainly those receiving a pension, showed more psychopathological features as monitored on MMPI than the patients without complaints. They also reported substantially less satisfactory occupational, family and social life than the patients considered to be complaint-free. Finally, they also differed in their modes of coping with their back problems. PMID- 2975066 TI - Signs and symptoms after chemonucleolysis. A detailed evaluation of 214 worker's compensation and noncompensation patients. AB - The effectiveness of chemonucleolysis was examined in a retrospective study of 214 consecutive patients with herniated lumbar discs. All patients were candidates for surgery. All underwent myelography, and most had computed tomography (CT) scans. The clinical status of patients was assessed at 6 weeks and at 6 months, and evaluation was made from responses of the patients to a questionnaire at 1 year. Evaluation at 1 year demonstrated that the results of chemonucleolysis were successful for 181 patients (84.6%). Of the 172 patients who received no worker's compensation, the procedure was successful for 90.7%. Of 42 with worker's compensation, the success rate was 59.5%. After chemonucleolysis, most patients showed neurologic improvement. Data comparing neurologic symptoms and signs between compensation and noncompensation patients are presented. This is the first report of objective data after chemonucleolysis in these two groups. A discussion on the morbidity and mortality of chemonucleolysis is included, along with a comparison of cost between chemonucleolysis and laminectomy. PMID- 2975067 TI - Neurophysiological investigation of effects of the D-1 agonist SKF 38393 on tonic activity of substantia nigra dopamine neurons. AB - The effects of the D-1 agonist SKF 38393 on tonic activity of rat substantia nigra pars compacta dopamine neurons were studied using extracellular, single unit recording techniques. Unlike nonselective D-1/D-2 dopamine agonists or the D 2 agonist quinpirole, SKF 38393 did not inhibit dopamine neuronal activity when applied iontophoretically or when administered intravenously in doses up to 20 mg/kg to chloral hydrate-anesthetized rats. Moreover, pretreatment with SKF 38393 did not alter the inhibitory response of these neurons to apomorphine or the D-2 agonist quinpirole. However, in locally anesthetized, gallamine-treated, artificially respired rats, dopamine cell activity was significantly altered by i.v. administration of SKF 38393; firing rate increases and decreases were observed. Administration of the inactive enantiomer of SKF 38393, S-SKF 38393, did not induce similar changes in parallel experiments. These results support the idea that unlike D-2 autoreceptor stimulation, D-1 receptor stimulation does not exert a direct local effect on dopamine neurons in the substantia nigra pars compacta and suggest that D-1 receptor stimulation at sites postsynaptic to the dopamine cells may indirectly affect the activity of some dopamine neurons through long-loop feedback mechanisms. PMID- 2975068 TI - Autoradiographic localization of 3H-paroxetine-labeled serotonin uptake sites in rat brain. AB - Paroxetine is a potent and selective inhibitor of serotonin uptake into neurons. Serotonin uptake sites have been identified, localized, and quantified in rat brain by autoradiography with 3H-paroxetine; 3H-paroxetine binding in slide mounted sections of rat forebrain was of high affinity (KD = 10 pM) and the inhibition affinity constant (Ki) values of various drugs in competing 3H paroxetine binding significantly correlated with their reported potencies in inhibiting synaptosomal serotonin uptake. Serotonin uptake sites labeled by 3H paroxetine were highly concentrated in the dorsal and median raphe nuclei, central gray, superficial layer of the superior colliculus, lateral septal nucleus, paraventricular nucleus of the thalamus, and the islands of Calleja. High concentrations of 3H-paroxetine binding sites were found in brainstem areas containing dopamine (substantia nigra and ventral tegmental area) and norepinephrine (locus coeruleus) cell bodies. Moderate concentrations of 3H paroxetine binding sites were present in laminae I and IV of the frontal parietal cortex, primary olfactory cortex, olfactory tubercle, regions of the basal ganglia, septum, amygdala, thalamus, hypothalamus, hippocampus, and some brainstem areas including the interpeduncular, trigeminal, and parabrachial nuclei. Lower densities of 3H-paroxetine binding sites were found in other regions of the neocortex and very low to nonsignificant levels of binding were present in white matter tracts and in the cerebellum. Lesioning of serotonin neurons with 3,4-methylenedioxyamphetamine caused large decreases in 3H paroxetine binding. The autoradiographic distribution of 3H-paroxetine binding sites in rat brain corresponds extremely well to the distribution of serotonin terminals and cell bodies as well as with the pharmacological sites of action of serotonin. PMID- 2975070 TI - [A bad back--so what?]. PMID- 2975069 TI - Interaction of a vasopressin antagonist with vasopressin receptors in the septum of the rat brain. AB - The ability of d(CH2)5-Tyr(Me)-arginine-8-vasopressin, an antagonist of peripheral pressoric (V1-type) vasopressin receptors, to label vasopressin binding sites in the septum of the rat brain was evaluated. Using crude membrane preparations from the septum, 3H-arginine-8-vasopressin (AVP) specifically labels a single class of binding sites with a Kd of 2.9 nM and maximum binding site concentration of 19.8 fmole/mg protein. 3H-Antag also labels a single class of membrane sites but with higher affinity (Kd = 0.47 nM) and lower capacity (10.1 fmole/mg protein) than 3H-AVP. The rank order of potency of various competitor peptides for 3H-AVP and 3H-Antag binding was similar. Oxytocin was 100-1,000 fold less potent than AVP in competing for binding with both ligands. 3H-AVP and 3H Antag showed similar labeling patterns when incubated with septal tissue slices. Unlabeled Antag also effectively antagonized vasopressin-stimulated phosphatidylinositol hydrolysis in septal tissue slices. PMID- 2975071 TI - [Clinical aspects of differential-diagnostic ECG analysis in practical electrocardiography]. AB - Individual assessment of ECG in any field of medicine requires, in the first place, analysis of clinical findings and, in the second place, the selection of a "specific" complex of symptoms from recorded ECG deviations reflecting cardiopathology proper (hypertrophy of different parts of the heart, a focal process of different layers, the extent and stages of disease development, SA, AV and intraventricular block, a certain type of disturbance of impulse formation, etc.). One should bear in mind the arbitrary concept of specificity of any ECG syndrome and its components applied specially to differential ECG diagnosis between different processes. When analyzing "nonspecific" ECG shifts one should necessarily consider a degree of reliability of one or another nonspecific sign for objective confirmation of supposed pathology which could probably cause (for clinicoanamnestic reasons) the appearance of this sign. PMID- 2975072 TI - [Early diagnosis of left-ventricular hypertrophy using precordial electrocardiographic mapping of the heart]. AB - Quantitative indices of instantaneous maps of the distribution of electric potential on the chest surface were compared to a left ventricular myocardial mass value, calculated on the basis of echocardiographic results and echocardiographic signs of left ventricular hypertrophy, in 77 patients with essential hypertension or neurocirculatory dystonia of hypertensive type and in 25 healthy controls. Four quantitative cartographic indices were shown to be very important for diagnosis: 1) tMaxk; 2) tapic; 3) + sigma QRS; 4) NQ15. The sensitivity, specificity and accuracy of diagnosis of left ventricular hypertrophy was over 80%. Cartographic quantitative indices were shown to correlate with a high degree of significance (p less than 0.01) (r much greater than 0.75) with a left ventricular myocardial mass value calculated on the basis of echocardiography results. PMID- 2975073 TI - ['Home care'--a second look]. PMID- 2975074 TI - [Low backache in nurses. Report of a study]. PMID- 2975075 TI - [Moving and lifting of patients and loads]. PMID- 2975076 TI - [Effectiveness of outpatient dental care for the schoolchildren of Arkhangelsk]. PMID- 2975077 TI - [Fluorosis and caries of the deciduous teeth]. PMID- 2975078 TI - [Experience with the methodologic guidance of intern training in pediatric dentistry]. PMID- 2975079 TI - [The teaching of the section on maxillofacial neoplasms at a dentistry department]. PMID- 2975080 TI - A toxin from Pasteurella multocida type D causes acute hepatic necrosis in pigs. PMID- 2975081 TI - Effect of treatment of first season calves with an OPRB on their immunity to lungworm in the second season. PMID- 2975082 TI - [Effect of naphthalane baths on the cardiovascular system in patients with osteoarthrosis deformans associated with ischemic heart disease]. PMID- 2975083 TI - [Many years of experience with a team method of work at a mud therapy clinic of the Saki health resort]. PMID- 2975084 TI - [Pretumorous states of the stomach in workers manufacturing organic glass]. PMID- 2975085 TI - [Immunity in patients with gastric and duodenal peptic ulcer (a review of the literature)]. PMID- 2975086 TI - [The thrombocytic beta-thromboglobulin level of patients with blastic leukemia and chronic myeloleukemia]. AB - The thrombocytic beta-thromboglobulin (beta-TG) level--a specific globulin secreted by the alpha-granules--is an important criterion in the contemporary diagnosis of acquired thrombocytopathies. The beta-TG was determined by the radioimmunologic test of the firm "Amersham" in thrombocytic lysates and thrombocyte-poor plasma of 54 persons: 24 patients with acute leukemia, 14 patients with chronic myeloid leukemia and 15 healthy controls. The leukemic patients were with a preliminary proved thrombocytopathy type "empty thrombocytic pool disease" which had been proved via aggregation measurement by the ATP and ADP levels in the thrombocytes and by the thrombocytic factor 4 level. While the intraplatelet beta-TG concentration in acute leukemia and chronic myeloid leukemia was found unchanged, in the patients with acute leukemia its secretion in the plasma was decreased (154.71 + 16.77 ng/10(5) platelets). The data interpretation shows that in these malignant hemopathies the alpha-granules do not take part in the "empty pool disease". In acute leukemia the pathogenesis of the thrombocytopathy is determined by the so-called "thrombocytic secretion paresis" which is confirmed by the thrombocytic factor 4 low level. PMID- 2975087 TI - [Use of the cholinolytic preparation Atrovent in high doses at short intervals in patients with chronic bronchial obstruction]. AB - The bronchodilating efficacy of two doses of Atrovent are studied: the first dose is of 60 mg applied by inhaling once only; the second dose is of 240 mg applied also by inhaling but in divided 4 single doses of 60 mg taken at one hour intervals. 14 asthmatic patients were examined--7 patients with atopic bronchial asthma and 7 patients with non-atopic form of the disease. To all patients the 240 mg dose of Atrovent was given. To 10 of the patients the single 60 mg dose of Atrovent was given at the same conditions but in another day. The larger dose exerts a better and more prolonged bronchodilating action. An optimal dose of 180 mg of Atrovent given by inhalation in divided doses of 60 mg at one hour intervals is recommended. The last 60 mg Atrovent of the 240 mg dose used in the experiment do not improve the immediate bronchodilation which is probably due to a blockade of the receptors by the drug. PMID- 2975088 TI - [Prune belly syndrome. Prenatal diagnosis and obstetric procedure]. AB - With an incidence of between 1 in 30,000 and 1 in 50,000 births, prune-belly syndrome (PBS) is a rare malformation syndrome. The phenotypical and pathoanatomic changes range from discrete expression to very severe malformation complexes with extremely poor prognosis. Malformations of the kidneys and the efferent urinary tract are common. In such cases, oligohydramnios often develops early in such fetuses, followed by Potter's sequence. This syndrome can generally be diagnosed by timely ultrasound investigations, i.e., before the end of the first half of pregnancy. Striking features in the fetus are often oligohydramnios and a greatly dilated urinary bladder, more rarely multicystic-dysplastic kidneys and other malformations of the urogenital tract. Diagnostic possibilities and obstetric procedure are described on the basis of ten cases seen between January 1984 and July 1987 at Heidelberg University Gynecological Clinic. PMID- 2975089 TI - [An enlarged neck fold: a sonographic marker of Down's syndrome]. AB - We report about one case of twin pregnancy where one fetus is healthy and the other has Trisomie 21. The one large skin fold on the neck is observe in the ultrasound. This ultrasound finds is noticed in the Ullrich-Turner-Syndrome as well as in the Down Syndrome. The Trisomie 21 was ascertained after the birth. The clinical meaning of this sonographic criterium will be discussed in this paper. PMID- 2975090 TI - [5-year survival rate in acute transmural heart infarct following thrombolysis and immediate coronary angioplasty]. AB - Two-hundred-eighty-six patients with acute transmural myocardial infarction underwent thrombolytic treatment (Streptokinase) between 1980 and 1986. In the earlier years patients were treated by thrombolysis only (n = 158) and in more recent years by thrombolysis followed by immediate percutaneous transluminal coronary angioplasty (n = 128). Age, sex, incidence of previous infarction and incidence of multivessel disease were comparable between groups. Patency of the infarct vessel (TIMI 3) was higher after thrombolysis combined with angioplasty than after thrombolysis alone (87% vs. 70%, p less than 0.001), and the residual stenosis of the infarct vessel was lower (46% vs. 84%, p less than 0.05). Hospital mortality (thrombolysis combined with angioplasty vs. thrombolysis alone) was 6% vs. 13%; one-year mortality was 8% vs. 21%, and five-year mortality was 18% vs. 31% (p less than 0.02). We conclude that treatment of patients with acute transmural myocardial infarction by thrombolysis combined with angioplasty is followed by a better long-term prognosis than treatment by thrombolysis only. PMID- 2975092 TI - [Simulation of a coronary vascular complication in transluminal coronary angioplasty by reversible intima folding]. AB - The injection of contrast material into the right coronary artery via a guiding catheter while a Harzler dilatation catheter had already been introduced revealed an inhomogeneous filling of the vessel suspicious of an extensive thrombosis or a dissection. On completion of the angioplasty and removal of the balloon catheter, the artery again filled homogeneously showing a good angioplasty result. It is concluded that a complication had only been faked during angioplasty due to intimal folding and subsequent flow obstruction following the insertion of the balloon catheter. PMID- 2975091 TI - [The cardioprotective effect of verapamil in acute percutaneous transluminal coronary angioplasty]. AB - Verapamil improved the ischemic tolerance of the myocardium in experiments in animals. Therefore, 20 patients were examined during percutaneous transluminal coronary angioplasty (PTCA) of a proximal LAD stenosis in order to evaluate the ability of verapamil to improve the ischemic tolerance in man. Before the second dilatation, 1 mg verapamil was given intracoronarily to 10 patients, the other 10 patients received placebo ic. Before and after each of the three inflations, blood samples were obtained from the coronary sinus in five patients of each group to analyze the contents of lactate, pyruvate, and pH. Verapamil caused a significant prolongation of the inflation from 58 +/- 12 s to 83 +/- 20 s. This effect was persistent also during the following inflation (96 +/- 19 s). The onset of angina was delayed (p less than 0.05). ST-wave elevations and T-wave amplitudes were smaller after verapamil, in spite of the increased duration of inflation, as compared with the control group. The time until ST elevations of 0.1 mV occurred was increased from 17 +/- 3 s to 57 +/- 18 s (p less than 0.05). The increase in lactate in coronary sinus blood was less pronounced after verapamil (48% of control; p less than 0.05). Intracoronary verapamil before PTCA of the LAD improved the ischemic tolerance of the poststenotic myocardium significantly as evaluated by measurements of electrocardiographic and metabolic parameters. No side effects occurred during the injection of verapamil into the left coronary artery. PMID- 2975093 TI - [Percutaneous angioplasty of the stenotic aortic valve: results, hemodynamic effects and complications]. AB - In 24 patients with aortic stenosis (13 men and 11 women, aged 47 to 80 years; mean age 67 years) a percutaneous angioplasty of the aortic valve was performed. The NYHA functional class improved in 20 patients. There was also a significant (p less than 0.01) decrease of the mean aortic gradient from 75.1 +/- 21.8 mm Hg to 45.3 +/- 11.7 mm Hg as well as an increase of the aortic valve area from 0.61 +/- 0.12 cm2 to 0.96 +/- 0.3 cm2. The end diastolic volume index decreased from 121.3 +/- 37.4 ml/m2 to 99.6 +/- 24.8 ml/m2 to the end diastolic volume index from 49.1 +/- 26.6 ml/m2 to 34.6 +/- 13.4 ml/m2 (both significant: p less than 0.01). The ejection fraction increased from 61.3 +/- 11.8% to 65.8 +/- 7.8% (p less than 0.05). Four patients presented a transient left bundle branch block. The corrected QT interval increased significantly (p less than 0.01) from 388.9 +/- 29 ms to 401.4 +/- 42.6 ms. Complications occurred in eight patients, being severe in three of them (one death from intractable internal hemorrhage, two cases of right sided hemiparesis). CONCLUSIONS: Percutaneous aortic valve angioplasty represents an alternative to the surgical procedure. In our patients there was not only an increase in the aortic valve area, but also an improvement in the ejection fraction. Nevertheless, unforeseeable complications may occur which obscure the results. In the future, widened experience and improved technique will reduce complications. PMID- 2975094 TI - [Rotation angioplasty--initial clinical results of peripheral vascular occlusions]. AB - After experimental investigations in postmortem human arteries, 19 patients with chronic peripheral artery occlusions were treated with a new technique between December 1986 and October 1987. In 17 patients the superficial femoral artery, and in two patients the popliteal artery, was completely occluded. The length of occlusions were between 5 and 25 cm (mean 10.9 cm); the duration (estimated according to patient's history) was 5-48 months (mean 17.2 months). In five patients, durations of up to 30 months had been documented by angiography. A flexible, blunt, motor-driven rotating catheter was introduced over an 8 or 9 F sheet and rotational angioplasty was performed at low speed, up to 200 rpm. In 11/14 patients in whom this new technique was used as the first attempt, the occlusions could be successfully reopened. In two patients after failure of the conventional technique the rotating catheter could not bypass the preexisting dissections in the same intervention. In 2/3 further patients after failure of the conventional technique the occlusions could be successfully reopened in a second intervention after several weeks. In none of our 19 patients did a perforation occur. It is concluded that by using the new technique, chronic peripheral artery occlusions can be reopened with a high success rate and without the danger of vessel wall perforation. The method can also be applied in patients in whom conventional techniques have failed. PMID- 2975095 TI - [Isradipine (PN 200-110), a new dihydropyridine calcium antagonist with slight negative inotropic properties in comparison with nifedipine]. AB - The influence of isradipine (PN 200-110), a new dihydropyridine calcium antagonist, in comparison to nifedipine and placebo on hemodynamics and left ventricular function was investigated in patients with coronary artery disease (10 patients in each group). The drugs were infused intravenously within 30 min in a dosage which led to a comparable afterload reduction (nifedipine [N] 2 mg, isradipine [I] 0.5 mg, AOP mean decrease: N, 14.7%, I, 13.1%). Increase of cardiac output (N +12.5%, I +15%) and decrease of systemic vascular resistance (N -29.2%, I -25%) were equal in both groups. Nifedipine caused a significant reflex increase of heart rate (+9.2%, p less than 0.001), which was not present with isradipine. The consequence was a significant decrease of the rate-pressure product with isradipine only (-12.5%, p less than 0.001) and not with nifedipine. Although afterload reduction was equal in both groups, isradipine caused a more pronounced decrease of LV volumes (EDVI: N -10%, I -16%) and an increase of ejection fraction (N +8%, I +14%). A significant increase of dp/dtmax, as a result of the afterload reduction, occurred after isradipine only (+13.5%, p less than 0.001) with no changes of dp/dtmax after nifedipine. Since changes of parameters (preload, afterload, HR, LVSP), which influence dp/dtmax independent of the inotropic state, were equal in both groups, the increase of dp/dtmax after isradipine should be a result of less negative inotropic properties of isradipine compared to nifedipine. PMID- 2975096 TI - [Elevated levels of atrial natriuretic peptide and plasma catecholamines in arterial hypertension--indications for an interaction]. AB - In this study plasma levels of atrial natriuretic peptide and of the catecholamines epinephrine and norepinephrine were investigated in hypertensive patients (HT) (n = 30). 22 normotensive patients (NT) served as controls. Hypertensives showed an elevated ANP-level in comparison with controls (46.8 +/- 3.3 vs. 36.8 +/- 3.3 pg/ml, M +/- SEM, p less than 0.01). When patients with myocardial infarction or with reduced ejection fraction were excluded, the same relation was demonstrated (49.3 +/- 3.2 vs. 33.6 +/- 2.0 pg/ml, p less than 0.01). Plasma norepinephrine was 230.8 +/- 52.3 pg/ml in HT compared with 138.0 +/- 19.6 pg/ml in NT (p less than 0.05). Epinephrine was 70.8 +/- 10.5 vs. 54.8 +/- 9.7 pg/ml in HT and NT. To exclude an increased left ventricular enddiastolic - and hence left atrial - pressure as the cause for the elevation of ANP and norepinephrine, HT and NT were matched for the same levels of enddiastolic pressure (LVEDP) (n = 18). For each level of LVEDP ANP was higher in HT than in NT (p less than 0.01). The same held true for norepinephrine (p less than 0.05) and to a lesser extent for epinephrine (p = 0.09). Our results demonstrate that patients with essential hypertension exhibit markedly elevated levels for ANP and catecholamines which is not due to myocardial failure. We propose that the increased secretion of the vasodilatory hormone ANP serves as counterregulation against the vasoconstrictor norepinephrine. The endocrine function of the heart may play a pivotal role in the modulation of sympathetic activity. PMID- 2975098 TI - [Results of thrombolytic therapy in acute myocardial infarct]. AB - Acute thrombosis of highly stenosed coronary arteries often results in acute myocardial infarction. The rationale for thrombolytic therapy is to restore blood flow before irreversible myocardial cell damage develops. In this review the results of more than 60 interventional studies in which reperfusion or "patency" rates, left ventricular function, infarct size, as well as intrahospital and long term mortality following thrombolytic therapy were assessed, are reported and analyzed with respect to the patterns of applying thrombolytic therapy (different thrombotic agents and percutaneous transluminal coronary angioplasty). The results show that a significant reduction in infarct size, improvement in left ventricular function, and decrease in early- as well as late-mortality can be achieved when successful recanalization is established within four hours after onset of symptoms. Thrombolytic efficacy with respect to patency rates is highest after intravenous application of so-called fibrin-selective thrombolytic agents (rt-PA) and APSAC (71% and 73%), intracoronary application of streptokinase (69%), and primary percutaneous transluminal coronary angioplasty (88%). By contrast, intravenous infusion of streptokinase results in recanalization of only 56% of occluded vessels. However, the improved reperfusion rates after treatment with rt-PA or intracoronary application of streptokinase are associated with a relatively high incidence of reocclusion (up to 20%). Patients' prognosis after successful initial thrombolysis is essentially influenced by additional interventions, e.g., percutaneous transluminal coronary angioplasty and surgical revascularisation. At present, several questions about the ideal thrombolytic agents or the optimal timing of additional interventions need to be clarified. Further studies are necessary to answer these questions. PMID- 2975097 TI - [Effect of the angiotensin-converting enzyme inhibitor ramipril on proteinuria, blood pressure and kidney function in histologically confirmed glomerulonephritis with nephrotic syndrome]. PMID- 2975099 TI - [Percutaneous transluminal coronary angioplasty following thrombolysis in acute myocardial infarct]. AB - The technique of combined medical and mechanical recanalization in acute myocardial infarction increases the reperfusion rate of occluded coronary vessels from 50% to 75% up to 90%. According to retrospectively performed analysis the reocclusion rate can be reduced at the same time from 17%-25% to about 7%-14%. The duration of occlusion of the coronary artery and the residual coronary stenosis following reperfusion are the main determinants of infarct size. The mechanical recanalization achieved by PTCA following thrombolysis improves the coronary flow and thus reduces infarct size, as demonstrated in experimental studies. Similar results can be expected in man as well. Indirect evidence has been demonstrated by ventriculographic and scintigraphic analysis of global and regional ventricular function. The time interval between successful thrombolysis and PTCA should be short, at least not longer than 5 days, because of the implications of residual stenosis for coronary flow and resulting infarct size and the incidence of reocclusion of the coronary vessel in the first few days following thrombolysis. In order to reduce the risk of reocclusion due to residual stenosis or residual coronary thrombi sufficient heparinization and anticoagulation is necessary. Cardiogenic shock in acute myocardial infarction represents a serious complication which is lethal in about 70%-80%. In these cases the technique of PTCA combined with thrombolysis may reduce mortality considerably to about 30%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975100 TI - [Operational assessment of the quality of cardiologic care as a means for raising the capacity of specialized medical services]. PMID- 2975101 TI - [Allergic occupational dermatoses in karakul sheep breeders]. PMID- 2975102 TI - [Argyria universalis]. PMID- 2975103 TI - [Familial favus]. PMID- 2975104 TI - [Pathogenesis of skin eruptions in psoriasis]. PMID- 2975105 TI - The social impact of childhood disablement. PMID- 2975106 TI - [Suppressor T cell function in undifferentiated collagenoses]. AB - In order to study early cellular defects in connective tissue diseases, suppressor T-cell function in undifferentiated connective tissue syndromes was determined. The capacity of T8(CD8)-positive cells to suppress pokeweed mitogen induced IgG and IgM secretion was studied in 14 patients. Compared with normals, a reduced activity of cells to suppress the IgM response was shown, whereas the suppression of IgG secretion was not affected. PMID- 2975107 TI - Suggested mode of action of D-penicillamine as an immunosuppressive agent in rheumatoid arthritis. AB - The mechanism by which D-Penicillamine is effective in the treatment of rheumatoid arthritis has been investigated. The results indicate that D Penicillamine in synergism with copper or ceruloplasmin in vitro inhibits the proliferation of T-lymphocytes and the activity of helper T-cells in supporting the generation of antibody-forming cells. This effect is mediated by the production of H2O2. The significance of these findings for in vivo processes is discussed. PMID- 2975108 TI - [OMR Prof. Walter Kunzel on his 60th birthday]. PMID- 2975109 TI - [Effect of smelting with propane-butane-oxygen and an acetylene-oxygen flame on the properties of Co and Ni base alloys]. PMID- 2975111 TI - [Cariogenicity of acidogenic and aciduric streptococci (S. mutans) in a rat experiment]. PMID- 2975110 TI - [Acidogenic properties of Streptococcus mutans]. PMID- 2975113 TI - [Status of the marginal periodontium in middle-aged adults. Strategies for care and organizational principles]. PMID- 2975112 TI - [Comparative morphogenesis of resorptive osteoid hyperregeneration of the periodontium and endodontium]. PMID- 2975114 TI - [Microbial composition of plaque as dependent on removable partial dentures]. PMID- 2975115 TI - [Composite and amalgam fillings in a 4-year clinical comparison]. PMID- 2975116 TI - [The author's experiences in the elevating and grafting of a free forearm flap in reconstructive measures of the face]. PMID- 2975117 TI - [Electrochemical corrosion behavior of dental alloys in various electrolytes]. PMID- 2975118 TI - [Production and inspection of "clean" surfaces for initial plaque formation studies]. PMID- 2975119 TI - [Abdominoplasty--personal surgical guidelines]. AB - An account is given by the authors of their own guidelines for surgical treatment of deformities of the anterior abdominal wall. The procedures described are based on experience obtained from 866 surgical interventions at the first author's clinic, between 1959 and 1987. Indications, techniques, and complications are discussed in some detail. PMID- 2975121 TI - [Treatment of long-term sterility by intra-tubal gamete transfer]. AB - It is reported about a first successful program of gamete intra fallopian transfer (GIFT) as a supplement to intrauterine insemination and to in vitro fertilization (IVF). In 29 patients after HMG stimulation of the ovaries and laparoscopic aspiration of follicles recovered oocytes have been introduced together with prepared sperm via a special catheter directly into the ampulla of the oviduct. In 21 patients with different causes of infertility optimal stimulation conditions could be achieved. A pregnancy rate of 41.4% is the result of 29 GIFT events and 3 of them resulted in spontaneous term deliveries (10.3%). PMID- 2975120 TI - [Acute hepatic porphyria in combination with configuration anomaly of the liver]. PMID- 2975123 TI - [The results of population-based psychopathologic research of paranoid schizophrenia]. AB - Paranoid schizophrenia was studied in 365 patients who constitute a nonrepresentative sample group inhabiting three urban districts. The population under study included groups with juvenile (31 patients), middle-aged (266) and late (68) onsets of the disease. Clinical variants of the disease were established with respect to its phenomenology and the course of delirium syndromes. PMID- 2975122 TI - [Pregnancy following gamete intra-fallopian transfer (GIFT)]. AB - In the present paper we will report on a pregnancy following a Gamete Intra Fallopian Transfer (GIFT). This is a 26 years old female, treated in our sterility dispensaire for a secondary sterility since 1982. All basic hormone levels were normal. The andrological examination of her husband showed an oligozoospermia stage 2. A following hormone treatment was unsuccessful. Stimulation, monitoring and laparoscopy are demonstrated. PMID- 2975125 TI - [The motives for the participation in work processes of severely mentally retarded persons at a psychoneurology boarding house]. AB - Population studied consisted of 727 oligophrenic patients aged 20 to 65 at the stages of pronounced debility and imbecility living in a special boarding-house. The degree of intellectual deficit proved to determine the patients' motives of participation in the labor activities: as the deficit increased, the part of socially mature motives diminished. The overall quantity of actual motives diminished with ageing. These motives completely disappear in imbeciles by 45. In pronounced debility the age-related increase in a part of motives caused by labor attractiveness was observed, in light imbecility the passive submissiveness increased. PMID- 2975126 TI - [Presentation of a case of spinal osteoid osteoma causing dorsal scoliosis. Role of high resolution CT]. AB - A case of vertebral osteoid osteoma in a 19 years old female is described. The tumor was located at D7 right peduncle and was responsible for a mild left convex curve dorso-lumbar scoliosis. The patient had been complaining for 3 years because of a stablike pain at medio-dorsal level. The symptomatology was exacerbated by erect position, during the night and was relieved by salicylic acid. Diagnosis was suspected on the basis of scintigraphic and computed tomographic features and confirmed at surgery. Limited value of plain films and the importance of bony scintigraphy and high resolution computed tomography to establish the localization and nature of the lesion are stressed. PMID- 2975124 TI - [Comparative effectiveness of the combined use of fenazepam, haloperidol, lithium and metabolic preparations in treating the psychopathologic disorders with obsessions in slowly progressive schizophrenia]. AB - The effectiveness of combined phenazepam, lithium, haloperidol and metabolic drugs (alpha-tocopherol, pyridoxal phosphate, nicotinamide) treatment of patients with slow progredient schizophrenia is analyzed. The psychotropic drugs were administered at doses lower below mean therapeutic ones by 1/2 to 1/3. Combined therapy proved effective even in the cases resistant to active antipsychotic treatment. No considerable untoward effects (extrapyramidal disorders) were found. The therapy had a differentiated impact on the obsessive syndrome and other borderline psychopathologic states. These advantages allow one to recommend the therapy for long-term support in schizophrenic patients with benign course of the disease. PMID- 2975127 TI - [Neurologic complications of measles in Parma in the period 1970-1986]. AB - In the years 1970-1986, six cases of encephalitis associated with measles were reported, two cases occurred in children living in chief town and four in other communes of the Province. The cases of measles reported in Parma were 6545 with an annual mean value of 385. The rate of complications over the reported cases was found to be 0.3 0/00. These overestimated value was re-elaborated taking into account the annual average of births in the chief town and in the Province, as in our Country, as well as in other European States, about 95% of children within ten years of age resulted to have been affected by measles. Thus the expected cases of measles resulted to be about 1500 in Parma and 3400 in the whole Province. The rate of complications now can be evaluated around the 0.07 0/00 for Parma and 0.1 0/00 for the Province. These epidemiological results confirm the importance of carrying out an effective vaccination program. PMID- 2975129 TI - [Hyponatremia during therapy with low doses of carbamazepine. Report on a clinical case]. AB - A case of a patient who developed hyponatremia both during normal dose and during low dose carbamazepine therapy is described. The clinical characteristics of hypo osmolar syndrome due to carbamazepine are similar to those of the syndrome of inappropriate antidiuretic hormone secretion. In literature, many cases of hyponatremia, related to assumption of normal dose carbamazepine, are reported; instead hyponatremia during low dose carbamazepine is rarely described. For this reason, a frequent monitoring of plasma sodium concentration, also in patients treated with low dose carbamazepine, is warranted. PMID- 2975128 TI - [Congenital stenosis of the mid ureter]. AB - Congenital strictures of the mid ureter of the left side are very rare entities. One case, recently diagnosed and treated, causing hydroureteronephrosis of the upper tract is herein reported. Few similar cases have been recorded in literature reviews: infact, on the right side, or in more cranial or caudal portions, a ureteral narrowing can be found more frequently and often due to aberrant vessels, underlying once more the actual rarity of this entity on the left side. In the present case of congenital stricture of the mid left ureter no extrinsic causes of narrowing were found at operation, confirming the presence of a topic and segmentary anomaly of the muscular portion of the ureteral wall. Surgical treatment led to complete recovery. PMID- 2975130 TI - [Localization of hydatid cysts in the left hemidiaphragm. Description of a case]. AB - The authors studied a 43-YR. old woman with a hydatid cyst, localized in the left diaphragm and extending into the left lung and the liver. De to difficulties in diagnosing the type, location, and extension of the lesion and its nonspecific symptoms, hydatid cysts are frequently not considered outside the endemic areas when formulating a differential diagnosis. Chest and abdominal CAT scanning permits a satisfactory definition of both the type of process and its topographical relationships. This permits the most appropriate surgical route to be chosen in performing a left thoracotomy in order to surgically afford the abdominal component of the cyst. This proved easy to remove, despite the existence of a tangle of adhesions, and had a favourable outcome, both immediately after surgery, and with long term follow-up. PMID- 2975131 TI - [Biopsy of the chorionic villi in the early diagnosis of chromosomal diseases]. AB - In the first trimester of pregnancy two simple chorionic villus sampling method, carried out under ultrasound guidance, with transabdominal or transcervical aspiration, was evaluated in 257 cases at risk for chromosomic diseases. The high efficacy of the transabdominal aspiration was demonstrated by obtain villus tissue for karyotyping in all but one case. In only one sample the abnormalities detected were not confirmed (one case of false positive). PMID- 2975132 TI - [Blood saving in heart surgery. Personal experience with 232 patients]. AB - In order to minimize the need of homologous blood transfusion during cardiac surgery four different blood sparing methods have been tested in four groups of patients. In 62 patients (group A) blood from operative field has been collected and processed by the Autotrans (Dideco (R)) apparatus. Intraoperative autotransfusion has been performed in 50 patients (group B). Prostacyclin (Pgh2) has been administered to 36 patients (group C), starting before systemic heparinization, and continuing during extracorporeal circulation. Standard techniques have been employed in a control group of 84 patients (group D). The following parameters have been examined: preoperative blood counts; length of extracorporeal circulation postoperative bleeding (including I.C.U); homologous blood volume transfused; and only in group A the mean number of blood units collected. Significant reduction of bank-blood consumption has been achieved only in B group (intraoperative autotransfusion) while in the remaining groups blood saving was not statistically significant. PMID- 2975133 TI - Percutaneous transluminal angioplasty of infrarenal arteries in intermittent claudication. AB - Percutaneous transluminal angioplasty was performed on 55 iliac and 31 femoropopliteal arteries in 71 patients with intermittent claudication (23 women, 48 men). The two-year patency rate was 80% after iliac and 41% after femoropopliteal angioplasty. In 17 femoropopliteal cases with lesions greater than or equal to 5 cm the 2-year patency rate was only 32%, but the corresponding figure for shorter lesions was 53%. Complicating haematoma appeared in 10% of the cases and the arterial state deteriorated in one patient. There was no distal embolization. Percutaneous transluminal angioplasty in intermittent claudication is indicated for all cases of occlusion or stenosis of the iliac artery and for occlusion or stenosis shorter than 5 cm of the superficial femoral or the popliteal artery. PMID- 2975134 TI - Finishing and polishing of composites. AB - Four different methods for finishing and polishing the surfaces of two different composite materials were studied by measuring the roughness of specimens at different stages of the procedures. Method 1 consisted of four different Shofu SuperSnap discs. Method 2 included the Shofu Dura-White stone, the Shofu CompoSite point, and the Vivadent polisher for composite resins. In method 3 the four 3M Sof-Lex XT discs were used. Method 4 comprised a green stone, a carbide finishing bur, and the Vivadent polisher for composite. All four methods were completed by using Kerr Command Ultrafine Lustre Paste. The composites used were Silux and P30. After the initial finishing, all four methods gave significant effect when used on the Silux, but on the P30 only methods 2, 3, and 4 gave a significant improvement. However, the degree of effectiveness of the methods varied greatly. PMID- 2975135 TI - [The child with hearing deficiency. Medical, educational, sociological and psychological aspects]. PMID- 2975136 TI - [A multidisciplinary approach to the management of deaf infants]. AB - An important epistemological change characterizes the education of hearing impaired and profoundly deaf children. The integration into ordinary schools and early home guidance are manifestations of this change. Early home guidance has been recognized as a promising new training approach for hearing impaired children. One breaks through the usual confinement in a specialized institution and strict individually based guidance. The family of the hearing impaired baby is directly involved in its early training. The authors have followed this new approach for several years now and explain their daily experience. An integrated training model is outlined in this article. PMID- 2975137 TI - Peripheral T-cell lymphoma with helper T-cell phenotype (Leu 3a+ Leu 8-). AB - Eleven cases of Leu 3a+ Leu 8- peripheral T-cell lymphoma (PTCL), excluding adult T-cell leukemia/lymphoma, were studied by immunostaining with monoclonal antibodies and enzyme histochemistry in order to clarify the histogenesis of PTCL. Seven of the eleven cases had varying degrees of polyclonal hypergammaglobulinemia. All cases were histologically characterized by neoplastic proliferation of clear cells and some cases showed a histologic background similar to IBL or AILD lesions with proliferation of immunoblasts or plasmacytoid cells and vascular proliferation. Immunohistologic analysis of PLP-fixed frozen tissues revealed that neoplastic clear cells expressed a Leu 3a+ Leu 8- phenotype (helper T-cell subset). The distribution of Leu 3a+ Leu 8- neoplastic cells corresponded closely to that of DRC-1+ cells, which are localized in the lymphatic follicles, but hardly at all with that of beta-glucuronidase+ vessels, termed PCV or HEV, which are usually present in T-cell areas. One case only progressed from Leu 3a+ Leu 8- IBL-like T-cell lymphoma (IBL-T), with proliferation of immunoblasts or plasmacytoid cells and vascular proliferation, to diffuse lymphoma of the large cell type showing none of these lesions. From these observations it is suggested that IBL-T might progress to T-cell-type monomorphous diffuse lymphoma. PMID- 2975138 TI - [Effects of gossypol acetic acid on heart mitochondrial membranes ATPase]. PMID- 2975139 TI - [Pharmacokinetic studies on 5-fluorouracil and its metabolite in rabbits by high pressure liquid chromatography]. PMID- 2975141 TI - Gene dosage effect in cells with monosomy of chromosome 22 derived from human meningiomas. AB - Enzymatic determinations of four enzymes, Arylsulphatase A (ARSA), alpha iduronidase (IDUA), beta-galactosidase (GLB) and hexosaminidase were performed in 19 cases of meningioma. Gene dosage effect was demonstrated for ARSA and IDUA; preliminary evidence for mapping a "protective protein" of GLB in the region 22pter----q11 was also obtained. PMID- 2975140 TI - Ultracytochemical investigation of calcium-activated adenosine triphosphatase (Ca++-ATPase) in chick tibia. AB - The ultrastructural distribution of Ca++-ATPase in bone cells of growing chick tibia was investigated by a cytochemical method in order to gain insight into possible sites of calcium ion translocation. Both osteoclasts and osteoblasts showed a polar distribution of reaction product along the plasma membrane. In osteoclasts, enzymatic activity occurred along the portion of the plasma membrane facing the marrow but not along the ruffled border or clear zone. The reaction product in these cells was due solely to Ca++-ATPase action. In osteoblasts, the plasma membrane facing away from bone (apical and lateral membrane) was very intensely stained, whereas the basal membrane was unstained. The reaction product in these cells appeared to be the result of both Ca++-ATPase and Ca++,Mg++ ATPase. In osteocytes, no plasma membrane staining was detectable. Mitochondrial staining in all three types of cells was more sensitive to fixation than was the plasma membrane enzyme, suggesting that mitochondrial and plasma membrane Ca++ ATPases are chemically distinct, as biochemical studies have shown. In general, mitochondria in osteoclasts stained more intensely than those in osteoblasts or osteocytes. Mitochondrial and vesicular sites of activity may be related to intracellular calcium storage, whereas calcium ATPases of the plasma membrane are presumed to be involved in calcium efflux from the cells. Calcitonin treatment did not alter the enzymatic distribution or intensity in osteoclasts. The striking polar distribution of both osteoclast and osteoblast plasma-membrane activity suggests that directional calcium pumping by these cells may be of importance in bone-forming and bone-resorbing mechanisms. PMID- 2975142 TI - Regulation of brain water and electrolyte contents: the opposite actions of central vasopressin and atrial natriuretic factor (ANF). AB - Considerable evidence indicates that regulation of the ionic environment of the brain is coordinated by a central neuroendocrine system capable of affecting the capillary endothelium, the choroid plexus, and the astroglia. All three cell groups are responsible for precise control of brain volume through adjustment of cell water and electrolyte content. With these considerations in mind, we have attempted to elucidate the possible involvement of the central vasopressin (AVP) and atrial natriuretic factor (ANF) systems in the regulation of the water and ion homeostasis of the brain tissue of rats: Vasopressin-positive vascular connections, investigated by immuno-electronhistochemistry, were found in close or direct contact with brain microvessels. Central administration of AVP (125 ng) or DDAVP (0.5 micrograms), with or without an accompanying water load, brought about a 1-1.3% water accumulation. Brain oedema caused by experimental subarachnoid haemorrhage had a different course in Wistar and Brattleboro DI rats, the latter being unable to synthetize AVP. These findings suggest that the centrally released AVP leads to brain water accumulation by increasing the water permeability of capillaries, and may facilitate the production of brain oedema in various pathological conditions. On the other hand, central administration of synthetic rat ANF (2 micrograms) prevented the water accumulation elicited in rat brain by systemic hypoosmolar fluid load, and led to a significant sodium loss from the nervous tissue by altering the capillary sodium permeability. The better understanding of these hormone receptors and their manipulations have exciting clinical implications. PMID- 2975143 TI - Alkylating toxins and the liver plasma membrane calcium pump/calcium atpase. PMID- 2975144 TI - The hepatic microsomal Ca2+ sequestering system. PMID- 2975145 TI - Regulation of liver plasma membrane Ca2+ pump. PMID- 2975146 TI - Purification of Ca2+-ATPase from rat pancreatic acinar plasma membranes using calmodulin-affinity chromatography. PMID- 2975147 TI - Is the current general anaesthesia for caesarean section really protective for the mother and the foetus? A study on beta-endorphin levels with different techniques of general anaesthesia. PMID- 2975148 TI - Rectus sheath block for diagnostic laparoscopy. AB - Sixty adult females of ASA grade 1 or 2 scheduled to undergo diagnostic laparoscopy were allocated randomly to one of two groups. In group A, laparoscopy was performed with a standardised general anaesthetic technique alone. In group B, the same general anaesthetic technique was supplemented with bilateral rectus sheath block. Postoperative analgesia was assessed at 1, 6 and 10 hours after operation. Visual analogue pain scores in group B were significantly lower than in group A despite a greater use of intramuscular analgesic injections in group A (p less than 0.005 in each case). PMID- 2975149 TI - Anaphylactoid reaction following the use of etomidate. AB - A 13-year-old female suffered urticaria and severe bronchospasm sufficient to cause hypoxic cardiac arrest after intravenous induction of anaesthesia. Etomidate was strongly implicated in the reaction. The management and mechanism of the reaction are described and discussed, together with consideration of future anaesthesia in the patient. PMID- 2975150 TI - Anaphylactic reaction after atropine. AB - A 38-year-old woman developed symptoms of anaphylactic shock after intravenous atropine and required adrenaline to maintain perfusion pressure. A strongly positive response was obtained on intradermal testing. The Prausnitz-Kuestner test was also positive, which indicated the presence of drug specific IgE antibodies. No response was obtained after hyoscine. PMID- 2975151 TI - Transient inability to see. PMID- 2975152 TI - Fiber sizes and histochemical characteristics of the rectus abdominis muscle of the rabbit under conditions of pregnancy and mechanically induced stress. AB - Histochemical properties and muscle fiber diameters of the rectus abdominis (RA) muscle of the female rabbit were compared with those of RA muscle of (1) pregnant females at term of pregnancy (30 days); (2) pregnant females in which the stretch on the abdominal wall natural to pregnancy was artificially prolonged with a 40 day period of intraabdominal appliance; (3) virgin females subjected to intraabdominal appliance for 30, 45, 60, and 70 days. The RA muscle of the control female rabbits is composed of 29.73% type I, 12.13% type IIA, 57.59% type IIB, and 0.54% type IIC fibers. The stimulation on RA muscle due to either normal pregnancy or implant in virgin females provoked changes in muscle fiber diameters only; with the normal pregnancy plus the subsequent period of mechanical stimulation, changes occurred not only in muscle fiber diameters, but there was also a significant increase in the percentage of type I fibers and a concomitant decrease of type IIA and IIB fibers. The fiber-type pattern in the RA muscle of male and female rabbit also was compared. The RA muscle of the male rabbit showed more type IIA and less type IIB fibers than found in that of the female. Type I fibers were larger in the female, but type IIA fibers were larger in the male. In addition, male rabbits also were subjected to 30, 45, 60, and 70 days of RA muscle stimulation by means of an intraabdominal appliance to ascertain if the influence of such stimulation upon the RA muscle differed between the two sexes. Some differences in the response of the male RA muscle were noted. PMID- 2975153 TI - Topical treatment of onychomycosis using bifonazole 1% urea/40% paste. PMID- 2975154 TI - Elevated free androgen index as an indicator of polycystic ovaries in oligomenorrhoea without obesity or hirsuties. AB - A group of oligomenorrhoeic women without obesity or hirsuties was investigated with high-resolution ultrasound, laparoscopy and biochemical parameters. In this series, polycystic ovaries (PCO), as defined by ultrasound and laparoscopy, are a common cause of oligomenorrhoea in women without the classic symptoms, and were strongly associated with an elevated free androgen index (FAI). Despite an elevated FAI, these women were not hirsute, It would seem reasonable to include a FAI in the investigation of the oligomenorrhoeic woman, along with the more 'standard' tests, such as thyroid function and a prolactin level. PMID- 2975155 TI - Salivary and plasma free testosterone and androstenedione levels in women using oral contraceptives containing desogestrel or levonorgestrel. AB - The effect of oral contraceptives (OC) containing a combination of ethinyloestradiol (EE2) and either desogestrel (Dg/EE2) or levonorgestrel (Lg/EE2) on plasma concentrations of sex hormone binding globulin (SHBG), total testosterone (T), free T, total androstenedione (A), free A and on salivary T and A concentrations have been studied. SHBG concentrations were higher in the Dg/EE2 group than in the Lg/EE2 group. Total T, measured by RIA with prior extraction and chromatography, is also higher in the Dg/EE2 group. Per cent free T was lower in the Dg/EE2 group. Plasma free T and free A concentrations were the same in both groups. However, free T and free A were significantly lower in the OC-groups than in controls. In contrast, by direct assay, no difference in total T was observed between the OC-groups. Since the per cent free T was lower in the Dg/EE2 group, it would be concluded that free T in the Dg/EE2 group is lower than in the Lg/EE2 group. Salivary free T and free A concentrations did not differ between control and OC-groups. There was good correlation between salivary and free hormone concentrations within each group. We conclude that the concentrations of androgens in saliva do not necessarily directly reflect their free concentrations in plasma. PMID- 2975157 TI - Polyamine transport in Neurospora crassa. AB - Polyamine transport in Neurospora crassa is concentrative and energy dependent in a dilute buffer. The saturable systems governing the uptake of putrescine (Km = 0.6 mM), spermidine (Km = ca. 0.24 mM), and spermine (Km = 0.07 mM) share components, as indicated by mutual inhibition among the polyamines. In addition, nonsaturable components prevail for putrescine and spermidine, particularly the former. Radiolabeled substrates, once in the cell, are released only slowly, even if unlabeled polyamines are included in the incubation medium. Permeabilization of cells with n-butanol leads to partial release of internalized 14C-polyamines, and the remainder is almost wholly exchangeable with added, unlabeled polyamine. Polyamine uptake was inhibited by the polyamines themselves and by a polyamine analog, methylglyoxal bisguanylhydrazone, but only weakly and incompletely by the basic amino acids arginine and ornithine. Uptake of putrescine and spermidine was inhibited by monovalent cations, Ca2+, and certain other components of the growth medium. As a result, uptake from the growth medium was very slow and largely by way of the nonsaturable uptake mechanism. PMID- 2975156 TI - Fermentation of peptides and amino acids by a monensin-sensitive ruminal Peptostreptococcus. AB - A monensin-sensitive ruminal peptostreptococcus was able to grow rapidly (growth rate of 0.5/h) on an enzymatic hydrolysate of casein, but less than 23% of the amino acid nitrogen was ever utilized. When an acid hydrolysate was substituted for the enzymatic digest, more than 31% of the nitrogen was converted to ammonia and cell protein. Coculture experiments and synergisms with peptide-degrading strains of Bacteroides ruminicola and Streptococcus bovis indicated that the peptostreptococcus was unable to transport certain peptides or hydrolyze them extracellularly. Leucine, serine, phenylalanine, threonine, and glutamine were deaminated at rates of 349, 258, 102, 95, and 91 nmol/mg of protein per min, respectively. Deamination rates for some other amino acids were increased when the amino acids were provided as pairs of oxidized and reduced amino acids (Stickland reactions), but these rates were still less than 80 nmol/mg of protein per min. In continuous culture (dilution rate of 0.1/h), bacterial dry matter and ammonia production decreased dramatically at a pH of less than 6.0. When dilution rates were increased from 0.08 to 0.32/h (pH 7.0), ammonia production increased while production of bacterial dry matter and protein decreased. These rather peculiar kinetics resulted in a slightly negative estimate of maintenance energy and could not be explained by a change in fermentation products. Approximately 80% of the cell dry matter was protein. When corrections were made for cell composition, the yield of ATP was higher than the theoretical maximum value. It is possible that mechanisms other than substrate-level phosphorylation contributed to the energetics of growth. PMID- 2975158 TI - Luminescence from the carbon monoxide derivative of Agaricus bispora tyrosinase. AB - The luminescence of the CO adduct of two isozymic tyrosinases isolated from Agaricus bispora, an edible white mushroom, has been studied. At room temperature the emission appears as a single smooth peak centered at 530 nm with FWHM of 2700 cm-1 and a lifetime of 36 microseconds. The lifetime and wavelength of the emission are virtually unchanged on lowering the temperature from 298 to 77 degrees K. Solvent composition affects the wavelength of emission minimally. The emission is quenched by oxygen but not by a series of substrate analogs, inhibitors, or Lewis bases. The emission further appeared independent of aggregation state of the enzyme or isozyme type. A comparison of these data is made with those obtained by other researchers for the tyrosinase from Neurospora crassa and for several hemocyanins. The comparison supports the hypothesis that regulation of enzymatic activity does not take place within the coordination sphere of the copper atom observed. In addition, it suggests that the 550- to 560 nm emissions previously observed may not be considered characteristic of all CO derivatives of coupled binuclear copper proteins. PMID- 2975160 TI - Lanthanum as a calcium-substituting ion for binding to sarcoplasmic reticulum ATPase. AB - Ca2+ binding and internalization in sarcoplasmic reticulum ATPase can be investigated by the use of La3+ as a Ca2+ analog. Displacement kinetics of Ca2+ bound by La3+ in native vesicles is a slow biphasic process (k1 = 0.55 s-1 and k2 = 0.05 s-1) that is consistent with the existence of two Ca2+ binding populations whereas in leaky vesicles there appears to be a single population (k = 0.57 s-1). Rapid quench experiments demonstrate that Ca2+ internalization occurs with an initial burst (approximately 8 nmol/mg protein) associated with the presence of a phosphate-donor substrate in the reaction medium. While acid quenching for measurements of phosphoenzyme is instantaneous, La3+ quenching allows completion of one catalytic and transport cycle due to the slow La3+ exchange with Ca2+. This explains the apparent inconsistencies in the kinetics and stoichiometry of phosphoenzyme formation and Ca2+ internalization that are observed under certain experimental conditions. PMID- 2975159 TI - Molecular characterization of four forms of phosphofructokinase purified from potato tuber. AB - Four forms of phosphofructokinase (PFK) have been purified to apparent homogeneity from tubers of potato (Solanum tuberosum cv. Record). Each had a final specific activity of about 200 mumol.min-1.mg-1 protein. Similar forms of PFK were found in partially purified extracts from tubers and leaves of other potato cultivars and related wild species. The relative molecular masses of three forms of PFK were about 200,000 whereas that of the fourth PFK was greater than 800,000. The four forms of PFK contained different proportions of four polypeptides which had apparent relative molecular masses of 46,300, 49,500, 50,000, and 53,000. These polypeptides gave different patterns of peptide fragments after chemical and proteolytic cleavage. Western blots and immunoprecipitation studies using antibodies raised against the individual polypeptides showed that all four are associated with PFK. Thus, potato tubers contain four distinct forms of PFK that differ in their subunit composition. PMID- 2975161 TI - [Phenotype changes in myocardial proteins in hemodynamic overloads]. AB - The phenotype of myocardial proteins is not set once and for all: it changes during ontogenesis, haemodynamic overload and in many other circumstances (e.g. dysthyroidism, diabetes). These changes are attributed either to the differential expression of multigenic protein families, or to modulation of single gene expression. In haemodynamic overload, this versatility of the myocardium enables it, usually by re-expression of its foetal phenotype, to find in itself the capacity for getting adjusted to new functioning conditions. The mechanisms that link the heart work overload to these phenotypic transitions, as well as their possible physiopathology, remain to be determined. PMID- 2975162 TI - [The force of evidence of quantitative morphological heart studies on normal slaughtered and transport-killed swine of the Schwerfurt meat breed (formerly line 150)]. PMID- 2975163 TI - Compensation status and symptoms reported by patients with chronic pain. AB - This study examined the initial symptoms of patients with chronic pain who were (n = 70) or were not (n = 52) involved in some aspect of the compensation system- worker's compensation, litigation, or Social Security Disability Insurance. Analyses indicated that compensation patients were discriminable from noncompensation patients (p less than 0.0001). Compensation patients were younger and less likely to be female; they also tended to report fewer surgeries, shorter pain durations, and more vocational and sexual disability. Finally, they perceived their medical conditions to be more severe than had been diagnosed by physicians. The groups did not seem to differ in severity of pain or psychologic distress. These data are consistent with studies indicating that compensation patients are not "symptom magnifiers," although the data do indicate that the life disruptions reported by these patients may be greater than those reported by patients not involved in compensation systems. PMID- 2975164 TI - Measuring the functional status of patients with low back pain. AB - For trials of back pain therapy and for many clinical purposes, the outcome of greatest interest may be patient functioning. Physical measurements of muscle strength and range of motion are only weakly correlated with actual patient behavior or symptoms, so the behavior and symptoms must be measured directly. Commonly used physician ratings on ordinal scales are often highly subjective and fail to distinguish particular activities which may improve from those which do not. This paper briefly considers the pressures for improving functional status measurement, describes five criteria for assessing and comparing functional questionnaires, and selectively reviews some newer instruments using these criteria. These newer instruments offer standardization of measurement, comprehensiveness, and generally good reproducibility and validity. Unfortunately, little is known about the responsiveness of these measures to small but clinically important changes. There is a need in this field to define the optimal balance between brevity on the one hand and comprehensiveness on the other, to assess and improve instrument responsiveness, and to compare instruments "head-to-head" to allow rational selection for particular applications. PMID- 2975166 TI - [The occurrence of Fusarium varieties and their mycotoxins in silo corn. 2. The formation of zearalenone in the field by artificial infection of silo corn with Fusarium culmorum (W. G. Smith) Sacc]. AB - The formation of zearalenone in a maize plot artificially infected with Fusarium culmorum was studied. The zearalenone concentration steeply increased only in the 8th week after inoculation and reached a maximum value of ca. 7 ppm, whereas zearalenone could not be detected in the control variants. The crude nutrient and dry matter content was not significantly influenced by the fungal infection. The infected crop showed average ear dry weights distinctly lower than that of the control variants (P less than or equal to 0.001). Apart from zearalenone, the mycotoxin deoxynivalenol was qualitatively detected in the infected maize. The toxicological relevance of the ascertained zearalenone content with regard to the health of dairy cattle and pigs was discussed. PMID- 2975165 TI - [The occurrence of Fusarium varieties and their mycotoxins on silo corn. 1. A method for the determination of zearalenone in corn and corn silage by high performance liquid chromatography (HPLC) with fluorescence detection]. AB - A method for the determination of zearalenone in maize and maize silage was developed which distinguishes itself by the effective and fast cleaning of the extracts with the help of a silica gel minicolumn. The samples were extracted with chloroform/methanol (9 + 1) and cleaned on a silica gel minicolumn after acid-base partition. The zearalenone was quantitatively determined optionally by means of high-performance liquid chromatography (HPLC) with fluorescence detection (excitation wavelength 236 nm, emission filter 418 nm) or thin-layer chromatography (TLC), p-methoxybenzene diazonium fluoroborate and aluminium chloride were used as detection chemicals. The limits of detection are 0.01 mg/kg (HPLC) and 0.1 mg/kg resp. (TLC), the average recovery is 81%. The method was used for the determination of zearalenone in grain maize, CCM silage and silage from whole maize plants. PMID- 2975167 TI - [The occurrence of Fusarium varieties and their mycotoxins in silo corn. 3. The effect of silaging on the zearalenone content of CCM corn]. AB - The influence of silaging on zearalenone already formed and on the growth of Fusarium culmorum in naturally contaminated CCM maize was studied. The zearalenone content remained approximately constant (13.35 +/- 2.94 mg/kg, n = 59) over the whole test period (12 weeks), whereas Fusarium culmorum could not longer be detected after 11 days. Thus the hypothesis that zearalenone survives the silaging process unchanged is experimentally strongly supported. The study confirms the view that zearalenone detected in maize silages is probably already formed in the field or during intermediate storing. Relations between the fermentation process and the toxin content of the silage could not be ascertained. PMID- 2975168 TI - Maternal mortality at the Women's and Children's Hospital, South Okkalapa (1978 1982). AB - The maternal deaths between the years 1978 and 1982 were studied. There were 22,468 maternities and 10,623 abortion patients treated at the hospital. There were 44 maternal deaths; 22 due to abortion and 22 due to other causes. The maternal mortality rate including abortions was 1.33 per 1,000 maternities and that excluding abortions 0.98 per 1,000. The abortion was 2.0 per 1,000 abortions treated at the hospital. To reduce maternal mortality, ways and means should be found to reduce the abortion deaths, most of which were avoidable. PMID- 2975171 TI - Interventional management of acute myocardial infarction. PMID- 2975172 TI - [Resolutions of the 101st meeting of the Executive Committee of the Panamerican Health Organization]. PMID- 2975170 TI - Enhancement of coronary conductance by alpha-human atrial natriuretic polypeptide without effects on myocardial contractility. AB - The effect of synthetic human atrial 28-amino acid peptide (alpha-human atrial natriuretic polypeptide, alpha-hANP) on coronary circulation and cardiac functions was examined in open-chest dogs. Intravenous injection of alpha-hANP increased coronary and systemic conductance, and coronary and aortic blood flow with a significant fall in blood pressure. Continuous infusion of alpha-hANP into the left anterior descending coronary artery (LAD) increased LAD blood flow in a dose-dependent manner. The linear regression analysis revealed the relationship of logit (changes in mean coronary conductance (delta MCC] = 1.45 x log (coronary plasma concentration of alpha-hANP) + 7.51 (r = 0.87, n = 29). REC50 of alpha hANP was 5.1 microM, where REC50 was the concentration to increase MCC to a half maximum MCC during reactive hyperemia after a 30-s coronary occlusion. alpha-hANP increased coronary conductance with no changes of myocardial oxygen consumption (MVO2) when blood pressure remained constant. Indices of myocardial contractility measured with a strain gauge arch, myocardial force (F), max dF/dt and LV max dp/dt, were not altered by either bolus intravenous injection or continuous intracoronary infusion of alpha-hANP. These results indicated a direct increase by alpha-hANP of coronary and systemic vascular conductance. PMID- 2975169 TI - The glycosylation properties of D2 dopamine receptors from striatal and limbic areas of bovine brain. AB - D2 dopamine receptors from bovine brain (caudate nucleus and olfactory tubercle) have been solubilized using sodium cholate/NaCl and their glycoprotein properties studied in terms of their interaction with wheat-germ agglutinin-agarose (WGA agarose). Under optimal conditions about 65% of the applied D2 dopamine receptors bound to WGA-agarose and could be eluted with N-acetylglucosamine. The ability of receptors to adsorb to the affinity column was shown to be dependent on the cholate and salt concentrations used. Digestion of the membrane bound D2 dopamine receptors with neuraminidase prior to solubilisation reduced the ability of the receptors to bind to WGA-agarose (50% of applied receptors bound) whereas digestion with N-acetylglucosaminidase did not significantly affect binding to WGA-agarose. Digestion with the two enzymes together resulted in a larger decrease in binding to WGA-agarose than was seen with the two enzymes alone (40% of applied receptors bound). Stepwise elution of bound receptors from the WGA agarose columns using 2.5 mM- and 100-mM-N-acetylglucosamine showed that about 40% of the bound receptors interacted with WGA-agarose in a low-affinity manner, the remainder showing a high-affinity interaction. Neuraminidase treatment reduced the low-affinity population suggesting that the interaction of oligosaccharides bearing sialic acid with WGA-agarose is of lower affinity and that higher-affinity binding is via N-acetylglucosamine. These data are discussed in terms of the heterogeneity of carbohydrate moieties on the D2 dopamine receptors within a brain region. In all the tests applied here, however, receptors from caudate nucleus and olfactory tubercle behaved identically so their glycosylation patterns must be very similar. PMID- 2975174 TI - [Present status of viral hepatitis in the world]. PMID- 2975173 TI - [Availability of courses in hospital pharmacy administration]. PMID- 2975175 TI - [Prevention of the problems of alcoholism]. PMID- 2975176 TI - On the preparation of dihematoporphyrin ether-free hematoporphyrin derivative. AB - A dihematoporphyrin ether-free hematoporphyrin derivative has been prepared by a base-catalysed dehydration of hematoporphyrin with sodium hydroxide. The identification was performed by HPLC and mass spectroscopy (FD-MS). The reaction of hematoporphyrin with 1 M sodium hydroxide for 24 h yields more than 90% of the monomeric porphyrins. PMID- 2975177 TI - Renovascular hypertension in moyamoya syndrome. Therapeutic response to percutaneous transluminal angioplasty. AB - Moyamoya ("puff of smoke") syndrome is a disease of children and young adults caused by fibromuscular dysplasia of the internal carotid and proximal cerebral arteries and is rarely associated with hypertension. We report a 34-year old woman with moyamoya syndrome who presented with symptomatic severe hypertension associated with unilateral renal artery stenosis. The patient underwent percutaneous transluminal angioplasty of the right renal artery with subsequent improvement in blood pressure control on reduced, and later, no antihypertensive therapy. This case illustrates that moyamoya syndrome may be an intracranial manifestation of a systemic arterial disorder. Renal artery stenosis may, in appropriate cases, be managed by percutaneous transluminal angioplasty. PMID- 2975178 TI - Synthesis of heparan sulfate proteoglycans by the isolated glomerulus. AB - Incorporation of [35S]sulfate into newly synthesized macromolecules was studied in the isolated rat glomerulus and found to be linear between 6 and 24 h. When whole glomeruli were treated under conditions that dissociate proteoglycan aggregates, greater than 90% of incorporated label was extracted. Of this, 80-90% was found to be the heparan sulfate proteoglycan. Similarly, a linear incorporation of [35S]sulfate into a glomerular basement membrane-enriched fraction was due almost entirely to proteoheparan sulfate. This predominance of heparan sulfate among the newly sulfated glycosaminoglycans has previously been observed in vivo and in the perfused kidney, but different patterns have hitherto been described in vitro. The present results suggest that under certain conditions, the isolated glomerulus is a suitable in vitro model for the study of proteoglycan synthesis. The pattern of incorporation of proteoglycans into the glomerular basement membrane reflects the time course and distribution of their synthesis by the whole glomerulus. PMID- 2975179 TI - Inhibition of myoblast fusion by bromoconduritol. AB - It has recently been reported that the glucosidase I inhibitor, N-methyl-1 deoxynojirimycin (MDJN), inhibits myoblast fusion whereas the mannosidase inhibitor, 1-deoxymannojirimycin (ManDJN), has no effect on fusion. We now report that bromoconduritol, which is an active-site-directed covalent inhibitor of glucosidase II, also inhibits fusion at concentrations that have no effect on the plating efficiency or growth of rat L6 myoblasts. Significant inhibition of fusion was obtained at concentrations as low as 50 micrograms of bromoconduritol/mL, whereas inhibition of cell growth did not occur until concentrations of 250 micrograms/mL were reached. Rat L6 myoblasts were grown in the presence and absence of processing inhibitors and were surface labelled with 125I. Analysis of the iodinated proteins by two-dimensional gel electrophoresis demonstrated that a number of high-molecular-weight proteins (greater than 90,000) detected at the surface of control cells were absent from the surface of cells treated with MDJN or bromoconduritol. It is suggested that MDJN and bromoconduritol prevent the translocation of these proteins to the cell surface. The high-molecular-weight proteins detected at the surface of control cells were also detectable in ManDJN-treated cells, indicating that inhibition of N-linked complex oligosaccharide formation does not affect the translocation of these proteins to the myoblast cell surface. PMID- 2975180 TI - Control of glycoprotein synthesis. The use of oligosaccharide substrates and HPLC to study the sequential pathway for N-acetylglucosaminyltransferases I, II, III, IV, V, and VI in the biosynthesis of highly branched N-glycans by hen oviduct membranes. AB - Glycoproteins isolated from hen oviduct contain highly branched asparagine-linked oligosaccharides (N-glycans). Six N-acetylglucosaminyltransferases (GlcNAc-T I, II, III, IV, V, and VI) are involved in initiating the synthesis of these branches, as indicated below: (formula; see text) where R is GlcNAc beta 1--- 4(+/-Fuc alpha 1----6)GlcNAcAsn-X. HPLC has been used to study the substrate specificities of these GlcNAc-T and the sequential pathways involved in the biosynthesis of highly branched N-glycans in hen oviduct. Oligosaccharides with free reducing GlcNAc termini were prepared from various glycoproteins by hydrazinolysis-re-N-acetylation and used as GlcNAc-T substrates and HPLC standards. Enzyme assay components were separated on AG1 x 8, followed by HPLC on amine-bonded silica columns eluted with acetonitrile-water mixtures. Absorbance at 195 nm and radioactivity of eluted compounds were monitored. Substrates and products were identified by comparison of their retention times with those of oligosaccharides with known structures. Enzyme assay by HPLC is more rapid and convenient than previous GlcNAc-T assays using lectin columns or electrophoresis. Since some substrates yielded multiple products, these could be used to assay more than one GlcNAc-T in the same incubation. GlcNAc-T VI was shown to act on both bisected and nonbisected GlcNAc-terminating tetraantennary oligosaccharide substrates; GlcNAc-T II, IV, and V acted poorly or not at all on bisected substrates. GlcNAc-T V was the only enzyme among the six transferases studied that could be assayed in the absence of Mn2+. PMID- 2975181 TI - [A comparison of blind and sighted children according to their oral health levels]. PMID- 2975182 TI - [Papillon-Lefevre syndrome (a case report)]. PMID- 2975183 TI - Comparison of four technetium-99m radiopharmaceuticals for detection and localization of gastrointestinal bleeding in a sheep model. AB - Four Tc-99 radiopharmaceuticals, Tc-99m sulphur colloid, Tc-99m red blood cells (RBCs), Tc-99m mercaptoacetyltriglycine (MAG3), and Tc-99m DTPA, were studied in an experimental animal model for detection and localization of gastrointestinal (GI) bleeding site in both the upper and lower abdomen. With Tc-99m sulphur colloid and Tc-99m RBCs, it was possible to detect and localize the GI bleeding site in the lower abdomen. With Tc-99m MAG3, it was possible to visualize the bleeding site in both the upper and lower abdomen. However, Tc-99m MAG3 is partially excreted by the liver into the bile, hence it will be difficult to use Tc-99m MAG3 to localize the GI bleeding site in the lower abdomen. With Tc-99m DTPA, it was possible to detect and localize the GI bleeding site simultaneously in both upper and lower abdomen. The overall background radioactivity was reduced considerably by diuresis with frusemide and catheterization of the urinary bladder. PMID- 2975184 TI - [Sleep apnea in severely handicapped patients: blood gas analysis and transcutaneous oxygen tension monitoring]. PMID- 2975185 TI - [Assessment and treatment of respiratory dysfunction in severely handicapped children--the efficacy of nasopharyngeal tubing]. PMID- 2975186 TI - Multiple gamma radiation sterilization of polyester fibres. AB - Gamma radiation with a dose of 2.5 Mrad has been found to be suitable to sterilize polyethylene terephthalate (PET) bulk materials intended for biomedical applications. The radiation stability of PET bulk materials and fibre may not however be taken as identical due to the changes in the polymer structure during the processing of bulk materials for fibre. The chemical changes occurring in PET fibres during single and multiple (prolonged) sterilization in air were investigated. It was found that single sterilization (2.5 Mrad) itself affected the PET yarn and fibre. This was exhibited by the increase of crystallinity from 30.5 to 37% in the case of yarn and from 40 to 44% in the case of fibre. The breaking load of the yarn also increased from 441 g to 451 g. These changes were attributed to the degradation of PET in the amorphous region and the recombination of degraded aliphatic segments. Sterilization at higher doses affected the crystalline region which decreased the crystallinity, breaking load and molecular weight. For samples irradiated at 2.5 Mrad, the breaking load increased, though the dispersity increased. This was attributed to cross-linking by recombination. The increase in crystallinity also enhanced the breaking load of the samples. Higher doses of sterilization led to drastic microstructural and macrostructural changes as seen from the molecular weight. It was inferred that the changes in crystallinity and microstructure that occurred during multiple sterilization might affect the biocompatibility of the material. PMID- 2975187 TI - Receptor characteristics and recovery of function following kainic acid lesions and fetal transplants of the striatum. II. Dopaminergic systems. AB - This experiment reports the development of striosomal-like patches in fetal striatal transplants grafted either into the intact, or kainic acid-lesioned, striatum of adult female rat brain. Although D2 receptor density approached control levels in the area of the patches, otherwise the transplanted striatum was relatively empty of dopamine receptors, and overall D2 development of the graft was strikingly reduced from controls. Transplants reversed lesion-induced hyperactivity under saline, but not amphetamine/apomorphine conditions, and led to an increased sensitivity of lesion and grafted animals following haloperidol injection. Transplants into the intact brain led to a lesion-like effect. M1 muscarinic cholinergic/D2 receptor density inversely correlated with behavior following amphetamine/apomorphine injections, while striatal cross-sectional area inversely correlated with activity after saline injection. These results suggest that abnormal maturation of the grafted striatum correlates with deficits in activity in grafted animals, and suggests that abnormal transplant development significantly impacts on transplant-induced behavioral changes. PMID- 2975188 TI - Decreased densities of dopamine D1 receptors in the putamen and hippocampus in senile dementia of the Alzheimer type. AB - The density and distribution of dopamine D1 receptors as labeled with [3H]SCH 23390 was analyzed in post-mortem brain tissue from patients with senile dementia of the Alzheimer type (SDAT) and in controls using quantitative autoradiography. In SDAT patients D1 receptor densities were markedly decreased in parts of the hippocampus, with reductions of up to 89% compared to the control values in the molecular layer of the dentate gyrus, 57% in the strata oriens and pyramidalis of the CA1 and 74% in the CA3 subfields. Significant decreases in D1 receptors were also observed in the putamen (23%) but not in the caudate and substantia nigra. A slight but not significant decrease of D1 binding was observed in most external layers of the temporal and occipital cortices. PMID- 2975190 TI - Dental treatment for mentally handicapped adults in general practice: parents' and dentists' views. PMID- 2975189 TI - [Death following administration of mithramycin]. PMID- 2975191 TI - Effect of sinus node on spontaneous release of natriuretic peptide in isolated atria. AB - Experiments were conducted to examine the release of atrial natriuretic peptide (ANP) in an isolated atrium in the presence and absence of sinus node tissue. The first series of experiments were conducted with the aid of a metabolic chamber to examine the spontaneous release of ANP by the right atrium with and without the sinus node region. The left atrium was also studied. The right atrium with the sinus node, quiescent right atrium without the sinus node, and the left atrium were incubated at 35 degrees C in 10 mL of oxygenated Tyrode's solution. After 40 min of equilibration, the incubation medium was removed at 10-min intervals for the determination of immunoreactive ANP concentration. The right atria with the sinus node released the highest amount of ANP into the incubation medium (32.2 +/ 2.7 pg.min-1.mg-1), compared with quiescent right atria (20.9 +/- 3.7 pg.min 1.mg-1). The left atria released the least amount of ANP into the incubation medium (9.9 +/- 1.5 pg.min-1.mg-1) when compared with the quiescent right atria and the right atria. In the second series of experiments, the right atrium was divided into the sinus node region and the quiescent right atrium, and these tissues were studied in paired fashion with a modified Langendorff preparation. The right atrium without the sinus node and sinus node region were perfused with Tyrode's solution, equilibrated with 95% O2 and 5% CO2 at 37 degrees C with a constant flow of 0.5 mL/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975192 TI - Atrial natriuretic factor alters autonomic interactions in the control of heart rate in conscious rats. AB - Regulation of heart rate was studied in rats receiving either i.v. saline at 64 microL/min or synthetic 28-residue rat atrial natriuretic peptide (ANF) at a dose sufficient to decrease mean arterial blood pressure by 10%. Autonomic influences were deduced from steady-state heart rate responses of each group to propranolol, atropine, or propranolol and atropine combined. A multiplicative model of heart rate control was used to derive quantitatively from the data the modulation of intrinsic heart rate by sympathetic and parasympathetic mechanisms. Animals receiving ANF showed a lower heart rate than control animals. This relative bradycardia was abolished by atropine. Blocking of sympathetic effects with propranolol had no effect on basal heart rate in either group, and atropinization led to significant increases in heart rate in both groups of rats. Mathematical analysis of the results showed that the bradycardia produced by ANF was due predominantly to a reduced intrinsic heart rate and to enhanced vagal inhibition of postganglionic sympathetic activity. Parasympathetic contribution to heart rate in the absence of sympathetic activity was negligible in control rats and small during ANF. We conclude that the major influences of ANF on heart rate control are a decrease of intrinsic heart rate and enhanced parasympathetic inhibition of postganglionic presynaptic sympathetic activity. PMID- 2975193 TI - Atrial natriuretic peptide interferes with calcium requirements in vascular tissues of the rat. AB - The inhibitory effect of atrial natriuretic peptide on the myotropic action of phenylephrine on superior mesenteric artery and thoracic aorta rings was studied to test the hypothesis that this peptide interferes with the mobilization of intra- or extra-cellular calcium produced by vasoconstrictor agents. In the absence of calcium in the bathing solution, phenylephrine (10(-6) M) produced a residual effect, which was antagonized in a concentration-dependent manner by the atrial peptide in both mesenteric artery and aorta rings. When calcium (2.5 mM) was added to the bathing solution after the response to phenylephrine in the absence of calcium, a further increase in the tonus of the tissue was observed. This effect was also antagonized by atrial natriuretic peptide in a dose dependent manner in the two tissues. These results suggest that atrial natriuretic peptide inhibits the effect of vasoconstrictor agents by functionally interfering with the mobilization of intra- and extra-cellular calcium produced by these vasoconstrictors. PMID- 2975194 TI - Markers of peripheral androgen action in vivo and in vitro. PMID- 2975195 TI - [Initial experience with the Burch colpopexy in the treatment of urinary stress incontinence]. PMID- 2975196 TI - Formation of a pyridinium derivative by reaction of 4-hydroxypentenal with glycine. AB - 4-Hydroxypentenal reacts with glycine in a slightly alkaline (pH 8.8) aqueous solution to 1-(1-carboxymethyl)-3-(2-hydroxypropyl)-pyridinium betaine. The yield of the pyridinium betaine isolated by preparative HPLC was 20 mol% and its structure was ascertained by 13C-NMR, IR, UV and mass spectroscopy. Several other products could be detected by HPLC in the 4-hydroxpentenal-glycine reaction mixture, their instability, however, impeded their preparative isolation. 4 Hydroxyalkenals are toxic products generated during lipid peroxidation and the results described explain in part the mechanism how these aldehydes react with nucleophilic amino groups in tissue. PMID- 2975198 TI - [The necessity for taking into account near visual acuity in the evaluation of body damage]. PMID- 2975197 TI - Streptozotocin-induced diabetes modulates the metabolic activation of chemical carcinogens. AB - The effect of chemically-induced diabetes on the hepatic microsomal mixed function oxidase system and the activation of chemical carcinogens was investigated in animals treated with streptozotocin (STZ). In order to distinguish between the effects of the diabetogenic chemical per se and that of the diabetic state, groups of STZ-treated animals received either nicotinamide simultaneously with STZ to prevent the onset of diabetes, or daily treatment with insulin in order to reverse the effects of diabetes. STZ-treated animals exhibited higher pentoxyresorufin O-dealkylase, ethoxy-resorufin O-deethylase, ethoxycoumarin O-deethylase, aniline p-hydroxylase and NADPH-cytochrome c reductase activities; similarly, increases were seen in cytochrome P-450 and b5 levels. All of these effects were prevented by nicotinamide and, at least partly, antagonised by insulin therapy. Treatment of animals with STZ markedly increased the activation, by liver microsomes in vitro, of Trp-P-1 and Trp-P-2 to mutagens, the effect being totally preventable by nicotinamide and successfully antagonised with insulin therapy. The diabetic animals were similarly more efficient in activating MeIQ but the effect was not preventable by nicotinamide or reversed by insulin. In contrast no changes were seen in the activation of IQ and only a modest increase in the case of MeIQx. It is concluded that diabetes may modulate the metabolic activation of some chemical carcinogens, presumably by changing the ratio of the various cytochrome P-450 isoenzymes. PMID- 2975199 TI - A comparison of dapsone with 13-cis retinoic acid in the treatment of nodular cystic acne. PMID- 2975200 TI - Palmo-plantar involvement in auto-immune blistering disorders--pemphigoid, linear IgA disease and herpes gestationis. PMID- 2975201 TI - Effects of changes in intravascular volume on atrial size and plasma levels of immunoreactive atrial natriuretic peptide in uremic man. AB - To study the trigger for the release of atrial natriuretic peptide (ANP) in man, we measured the atrial areas (AA) by 2-D echocardiography, the total blood volume (TBV) by 131I-serum albumin and plasma immunoreactive ANP (i-ANP) concentrations by radioimmunoassay, after prior plasma extraction, for 10 dialyzed uremic patients. Measurements were made when the patients were volume-loaded or volume depleted by isoosmotic ultrafiltration and again 48 h later, when they were again volume-loaded. Analysis of plasma extracts by high-performance gel permeation chromatography revealed that the greatest amount of the i-ANP fraction was a peptide eluting like human synthetic alpha-ANP. Ultrafiltration consistently decreased the TBV, while spontaneous regain of body-fluids caused TBV to rise to pre-ultrafiltration levels. Changes in TBV were closely related in time to changes in both right (RAA) and left (LAA) atrial area and in plasma i-ANP concentrations. Significant direct relationships were found between TBV and RAA, TBV and i-ANP and between both LAA and RAA and i-ANP. Furthermore, the decreases and the increases in TBV, RAA and LAA were closely correlated with changes in i ANP. Multiple regression analysis, however, revealed that the changes in plasma i ANP were mainly related to the changes in RAA, with little or no relationship to the changes in TBV or LAA. These findings are evidence for a positive feed-back between the level of intravascular filing volume, extent of atrial distention and amount of i-ANP released into the blood stream. PMID- 2975202 TI - B-cell-derived human interleukin 1. AB - This review addresses the questions of the molecular nature and of the physiological role of interleukin-1 (IL-1)-like activities produced by B lymphocytes. IL-1 was originally described as an exclusive product of activated monocytes/macrophages. The recent cloning of two genes for IL-1 (IL-1 alpha and beta), together with the availability of specific antibodies to these two species of IL-1 have allowed their identification as secretory products of a number of other cell types, including B cells. B cells secrete a variety of other autostimulatory factors and of IL-1-like molecules, the identification of which is still pending. In addition, B cells express receptors for IL-1, which has been shown to enhance proliferation and immunoglobulin synthesis. An important issue is that of whether B-cell-derived IL-1 serves a purpose in the physiology of the immune response. Inasmuch as IL-1 is required for T-cell response, it has been suggested that B-cell-derived IL-1 may contribute to the amplification of the immune response, particularly where B lymphocytes serve as antigen-presenting cells. PMID- 2975203 TI - Effects of prolonged picotamide therapy on platelet activity in patients with peripheral arterial disease. AB - An open study was carried out in 14 patients with peripheral arterial disease to investigate the effects of prolonged therapy with picotamide on platelet activity. Patients received daily oral doses of 900 mg picotamide for 1 month, 600 mg per day during the second month and 300 mg per day from the third to the sixth month of the study. Measurements were made before and during therapy of blood coagulation parameters and factors influencing platelet function, i.e. plasma beta-thromboglobulin and serum thromboxane B2. The results showed that there were no significant variations in platelet count, prothrombin time, partially activated thromboplastin time, presence and amount of fibrinogen in blood, and antithrombin III. Examination for fibrinogen degradation products was constantly negative and unaltered during therapy. Although plasma beta thromboglobulin values did not vary significantly, there was a significant and progressive reduction throughout treatment in serum levels of thromboxane B2. PMID- 2975204 TI - Comparison between oral procaterol and salbutamol in patients with bronchial asthma. AB - The efficacy of procaterol, a new beta 2-selective sympathomimetic drug, was compared with that of salbutamol and placebo in a double-dummy crossover study in 20 asthmatic patients. Procaterol (0.1 mg orally) was given twice daily and salbutamol (4 mg orally) 3-times a day. The study was made up of four consecutive 4-day treatment periods including two periods of plain placebo. A significant direct bronchodilating effect of both procaterol and salbutamol could be seen in PEF values, measured 4-times a day, compared with the effect of placebo (p less than 0.01 for both). Procaterol was slightly superior to salbutamol. The afternoon and evening PEF values during the procaterol period did not differ from the values during the placebo period. In symptom scores, there was significantly more tremor during the procaterol period than during the placebo period (p less than 0.01). Both procaterol and salbutamol produced more palpitation than placebo (p less than 0.05). The study shows that oral procaterol is a potent bronchodilator. The doses of procaterol and salbutamol were not equivalent. Procaterol with the dose used in the study was more potent. Despite this, the duration of the bronchodilator effect of procaterol on a twice daily dosage did not seem to be long enough in all patients. PMID- 2975205 TI - Soluble CD23/BLAST-2 (S-CD23/Blast-2) and its role in B cell proliferation. PMID- 2975206 TI - AIDS victims knock at door. PMID- 2975207 TI - Dentists wear gloves, balk at treating victims. PMID- 2975208 TI - Dental discrimination? PMID- 2975209 TI - Greene clinic provides care for Los Angeles AIDS victims. PMID- 2975210 TI - [Rate reduction of wall abscesses in daily abdominal surgery by repeated lavage with solutions of polyvidone iodine. 4006 cases]. PMID- 2975211 TI - Community outreach: Part II of a series. PMID- 2975212 TI - Trospectomycin, a novel spectinomycin analogue: antibacterial activity and preliminary human pharmacokinetics. AB - Trospectomycin (TSP; U-63366F) is a novel spectinomycin (SP) analogue with broad spectrum antibacterial activity. The in vitro activity of the analogue was compared to that of SP against approximately 400 bacterial isolates. The in vivo activity of the compound was assessed using experimental infection models for both Gram-positive and Gram-negative facultative bacteria. The preliminary human pharmacokinetics of TSP were evaluated following single-dose i.v. or i.m. administration. TSP was more active in vitro than SP (2 to 32-fold) against strains of numerous bacterial species, including staphylococci, streptococci, Haemophilus influenzae, Branhamella catarrhalis, Neisseria gonorrhoeae, Proteus spp., Bacteroides spp., Gardnerella vaginalis and Chlamydia trachomatis. The activity of TSP for most species of the family Enterobacteriaceae was comparable to that of SP. TSP was more active than SP (2 to 32-fold) in curing experimental infections due to streptococci, Salmonella typhi, Serratia marcescens, Klebsiella pneumoniae and Haemophilus influenzae. TSP was well-absorbed following both i.v. and i.m. administration. Pharmacokinetic analysis of microbiological assay data for the 1000 mg dose yielded the following mean values for the i.v. and i.m. routes, respectively: Cmax = 81.2, 28.7 micrograms/ml; serum half-life = 2.2, 2.2 h; Tmax = 25, 75 min; and AUC = 156.6, 116.2 h micrograms/ml. Pharmacokinetic analysis of assay data derived using the more sensitive HPLC assay revealed the biphasic nature of trospectomycin elimination, highlighted by a short apparent serum half-life (2.2 h) and a prolonged tissue half-life (approximately 36 h). TSP inhibits a variety of clinically important organisms, including agents of sexually transmitted diseases and pelvic inflammatory disease, and demonstrates favourable pharmacokinetic properties.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975213 TI - Pruritic rash with actinic keratosis and impending exfoliation in a patient with hypertension managed with minoxidil. AB - Dermatological toxicity has been reported following initiation of therapy with minoxidil, but no cases have been reported following prolonged use. We report the emergence of an erythematous weeping rash with impending exfoliation three years after the initiation of minoxidil therapy. Minoxidil was withdrawn and the patient responded to therapy with topical corticosteroids. Following minor surgery, the patient was inadvertently rechallenged with minoxidil. Within 24 hours of exposure bullous lesions reappeared in the extremities which again resolved with topical corticosteroids. Dermatological lesions observed on this patient were similar to those reported following acute minoxidil exposure and strongly implicate chronic minoxidil therapy. PMID- 2975214 TI - Left ventricular hypertrophy. Epidemiological insights from the Framingham Heart Study. AB - Left ventricular hypertrophy (LVH) has assumed an important role in clinical medicine as a result of the clinical implications of this often asymptomatic finding. Epidemiological data from the Framingham Heart Study have permitted an examination of prevalence, incidence, underlying predisposing factors and prognosis of LVH. Although LVH is an infrequent finding on the electrocardiogram, it is a forerunner of coronary disease, congestive heart failure, stroke and even peripheral arterial disease. Despite being strongly related to hypertension, LVH remains associated with excess risk for adverse cardiovascular morbid and fatal outcomes, even after adjusting for blood pressure. The risks associated with LVH are comparable with those of myocardial infarction. The recent introduction of echocardiography at the Framingham Heart Study has permitted the development of new criteria for LVH based on M-mode determined left ventricular mass. Unlike its electrocardiographic counterpart, echocardiographically determined LVH is a common finding, occurring in over 15% of the general population. Echocardiographic LVH is related to hypertension, obesity, valvular heart disease, coronary disease and advancing age. Ambulatory ECG results in subjects with echocardiographic LVH demonstrate increased risk for ventricular arrhythmias, which have been shown in other clinical settings to predict risk for sudden cardiac death. Preliminary data from Framingham and elsewhere suggest that echocardiographic LVH is associated with increased risk for cardiovascular disease morbidity and all-cause mortality. PMID- 2975215 TI - Left ventricular hypertrophy and antihypertensive therapy. AB - Cardiac adaptation to long-standing arterial hypertension consists of left ventricular hypertrophy (LVH), usually of the concentric type, i.e. an increase in wall thickness at the expense of chamber volume. LVH can no longer be considered only as a simple adaptive myocardial process; it drastically increases the risk of sudden death and cardiovascular morbidity and mortality, irrespective of the levels of arterial pressure. Patients with LVH have more premature ventricular contractions than patients without LVH or normotensive subjects, which indicates that LVH per se increases ventricular ectopic activity. Antihypertensive therapy should not only lower blood pressure, but also prevent or improve end-organ damage and therefore allow left ventricular mass to regress. Although they lower blood pressure, certain antihypertensive agents such as the thiazide diuretics and arteriolar dilators (hydralazine, minoxidil) have little or even a detrimental effect on LVH. In contrast, other agents such as angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, antiadrenergic drugs, and certain calcium antagonists decrease left ventricular mass in parallel with arterial pressure. Recent evidence has shown that a decrease in left ventricular mass induced by certain antihypertensive drugs suppresses ventricular ectopic activity by 85%. In contrast, left ventricular mass and ventricular ectopic activity remain unchanged or may even increase in patients treated with diuretics. It is not known whether the risk of sudden death can be decreased and the ominous prognosis of LVH altered by such specific antihypertensive therapy. PMID- 2975217 TI - Atrial natriuretic peptide in kidney of renal disease patients and healthy persons. AB - Regulation of renal excretion of atrial natriuretic peptide (ANP) was studied in kidney disease patients and healthy kidney donors. The measured ANP concentration in the patient's plasma did not correlate with their creatinine clearance (Ccr), while the fractional excretion of ANP (FEANP) significantly correlated with Ccr. FEANP in healthy persons is less than 1%. In the healthy donors of kidneys for transplantation, approximately 80% of the plasma ANP from the renal artery appeared in the renal vein. From these results, this high recovery of ANP in the veins does not appear to be adequately explained by its degradation in the renal arterioles and nephrons. The FEANP from kidney disease patients significantly correlated with FENa, FEK and FEP, but not with FECa and FEMg. The manner of ANP handling in the nephron may possibly differ from that of Ca or Mg. PMID- 2975218 TI - Comparative in vitro activity of A-56268. AB - The comparative in vitro activity of A-56268 was studied using 1,006 clinical isolates including streptococci, enterococci, staphylococci, Neisseria gonorrhoeae, Haemophilus influenzae and anaerobes. A-56268 showed activity comparable to that of erythromycin and was more active than josamycin and roxithromycin against erythromycin-sensitive aerobic and facultatively anaerobic gram-positive cocci. A-56268 was the most active macrolide against Clostridium spp. and Neisseria gonorrhoeae. Josamycin was more active than either A-56268 or erythromycin against the anaerobic gram-positive cocci and the Bacteroides fragilis group. Staphylococci moderately resistant or resistant to erythromycin (MIC 3.12-50 mg/l) remained susceptible to josamycin but not the other macrolides. PMID- 2975219 TI - Clinical experience with 99mTc-MAG3, mercaptoacetyltriglycine, and a comparison with 99mTc-DTPA. AB - The preparation, application and clinical usage of 99mTc mercaptoacetyltriglycine, MAG3, a tubular secreted compound, is described in the first 225 patients in a phase III study. Image quality, relative renal function, and renal transit times were compared with a 4 fold greater administered activity of 99mTc-DTPA in 11 patients. Correlation coefficients of 0.94 for relative function, 0.83 for parenchymal transit time index and 0.82 for whole kidney transit time index were found. Frusemide responses were similar. 99mTc-MAG3 is an efficacious radiopharmaceutical for routine renal radionuclide studies, giving excellent image quality in patients with hypertension, poor renal function, obstructive nephropathy or a renal transplant. PMID- 2975221 TI - Sequentially alternating hormone chemotherapy with high-dose medroxy-progesterone acetate and low-dose epirubicin for the treatment of hormone-resistant metastatic prostatic cancer. AB - Thirty patients with hormone-resistant metastatic progressive prostatic carcinoma were treated with sequentially alternating hormone chemotherapy. They were given 1,000 mg medroxyprogesterone acetate (MPA) orally for 26 days followed by intravenous doses of 25 mg/m2 epirubicin weekly for 4 weeks. The median duration of the treatment was 29 weeks (range 8-84). In 2 patients a more than 50% reduction in the size of measurable lymph node metastases was observed and in 2 others skeletal metastases decreased. Serum acid phosphatase normalized in 6 patients. Twenty-five patients achieved a subjective response (median duration 24 weeks; range 4-76 weeks). Median survival from the start of treatment (30 +/- 16 weeks) was unrelated to the achievement of subjective response. Normalization of serum acid phosphatase and a more than 50% reduction in serum alkaline phosphatase correlated with the achievement of a subjective response. Toxicity was generally mild, but in 1 case therapy was discontinued because of suspected cardiotoxicity. Sequentially alternating high-dose MPA low-dose epirubicin hormone chemotherapy has a marginal objective effect but a good subjective effect on progressing hormone-resistant prostatic cancer. PMID- 2975220 TI - Hormonal pattern and testicular histology in patients with prostatic cancer after long-term treatment with a gonadotropin-releasing hormone agonist analogue. AB - Seven patients suffering from prostatic cancer were treated with a depot form of D-Trp-6-luteinizing hormone-releasing hormone (LH-RH), a LH-RH agonist analogue (3 mg i.m. every 28 days) for a period of 24-32 months. The peptide induced a sharp and long-lasting inhibition of both gonadotropin and testosterone secretion. A sustained suppression of pituitary and testicular function was observed 40 days after the treatment was suspended. Testicular biopsies performed in all patients showed a marked impairment of Leydig cell mass and a complete spermatogenic arrest with a tubular derangement and fibrosis. Results indicate that the long-term continued gonadotropin-releasing hormone agonist analogue therapy induces not only a functional inhibition of testicular androgenesis but also anatomical testicular damage whose reversibility does not seem to be probable. PMID- 2975223 TI - Clinical studies on cell-mediated immunity in patients with urinary bladder carcinoma. Depression of cell-mediated immunity by adherent cells. AB - Lymphocyte blastogenesis by stimulation with phytohemagglutinin was compared in 10 control subjects and 20 patients with bladder cancer. Blastogenesis was measured by the incorporation of 3H-thymidine into lymphocytes using microplate assay. As a result, blastogenesis was significantly decreased in high stage patients compared to control subjects and low stage patients. When the nylon wool adherent cells were removed, lymphocyte blastogenesis was increased in high stage patients, while it was decreased in control subjects and low stage patients. This study provides evidence that in most bladder cancer patients with in vitro immunodepression, adherent suppressor cells are detectable, and that these cells may mediate the noted immunodepression. PMID- 2975222 TI - Total androgen blockade in advanced prostatic cancer. Critical review and personal experience. AB - The theoretical basis of 'total androgen blockade' in the treatment of advanced prostatic cancer is reviewed. The results of experimental and clinical data in the literature comparing medical or surgical castration alone versus castration plus peripheral antiandrogens are discussed. Most of these studies could not reproduce Labrie's results and do not establish the benefit of total androgen blockade. The author produces his own experience with LHRH analogs alone (Depot Zoladex; 30 patients) compared to orchidectomy plus antiandrogens (17 patients). After 2 years, there is no difference in the progression and death rates between the two groups. Moreover, the administration of an antiandrogen in 15 patients relapsing after first-line hormonal treatment (Zoladex or orchidectomy) produced an objective response in only 2 of them. 9 patients experienced a subjective response but the duration of response was very short (medium 3 months). PMID- 2975216 TI - Factors involved in the pathogenesis of hypertensive cardiovascular hypertrophy. A review. AB - All tissues can rapidly adapt their structural design whenever prolonged changes of load/activity occur within the limits characteristic of each tissue. This structural adaptation, however, is modified by various genetic and trophic influences. When antihypertensive therapy is considered in the hypertensive patient, such changes are usually well established and the cardiovascular system is structurally adapted to maintain a higher pressure than normal. Increased blood pressure and afterload cannot solely explain the development of cardiac hypertrophy. Permissive actions from the sympathetic nervous system and the circulating angiotensin II are likely, but conflicting results still exist. There is evidence for a functional renin-angiotensin system in the heart, which may be involved in the genesis of left ventricular hypertrophy. Also, a soluble factor in the hypertrophied myocardium that stimulates protein synthesis may play a key role in modulation of myocardial structure during development or regression of myocardial hypertrophy in hypertension. Hypertrophy of both the large and smaller arterial vessels has been shown to follow the same general pattern of development and regression as in the heart. The vascular hypertrophy (predominantly of the media in the arterioles) can be considered as the ultimate structural factor behind the progression of hypertension independent of the initiating factor. A vicious circle with the increased resistance as the key factor can be identified. There are at least 3 possible initiating factors: a small rise in arterial pressure, an abnormal or reinforced response to pressure, or trophic/mitogenic stimuli acting directly on the vascular smooth muscle cell. The ultimate goal in the treatment of high blood pressure is to reduce hypertension-related morbidity and mortality. Normalisation of structural cardiovascular changes is also important and easier to evaluate in the individual. Understanding of the pathogenesis of structural adaptation may help in the selection of the best treatment. PMID- 2975224 TI - 5-HT2 antagonists and minaprine block the 5-HT-induced inhibition of dopamine release from rat brain striatal slices. AB - 5-Hydroxytryptamine (5-HT) inhibited the K+-induced [3H]dopamine [( 3H]DA) release from slices of rat striatum. Minaprine (3-(2-morpholinoethylamino)-4 methyl-6-phenylpyridazine) attenuated the inhibitory effect of 5-HT in a dose dependent manner. 5-HT2 receptor antagonists, ketanserin and mianserin, prevented the effect of 5-HT as well as minaprine did. The inhibitory effect of 5-HT was not mimicked by a 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), and was not prevented by a 5-HT1A and 5-HT1B mixed receptor antagonist, propranolol. Minaprine was a potent inhibitor of the binding of [3H]ketanserin to binding sites in the striatum over the concentration range 10( 6)-10(-4) M. Lesion of the medial forebrain bundle with 6-hydroxydopamine (6 OHDA) significantly reduced the K+-induced [3H]DA release from the striatum and release was no longer inhibited by 5-HT. Lesioning, however, did not change significantly the [3H]ketanserin binding in the striatum. These results suggest that minaprine suppresses the inhibitory effect of 5-HT on DA release in the striatum via the inhibition of 5-HT binding at the 5-HT2 receptor on the nerve terminal of the DA-ergic neuron and, further, that the proportion of the 5-HT2 receptor site which is located on the nerve terminal of the DA-ergic neuron is small in the striatum. PMID- 2975226 TI - Differential effects of long-term antidepressant treatments on 8-OHDPAT-induced increases in plasma prolactin and corticosterone in rats. AB - Intravenous administration of 8-hydroxy-2(di-n-propylamino) tetralin (8-OHDPAT) to rats produced increases in plasma prolactin and corticosterone concentrations. Long-term or short-term treatment with the MAO type A inhibiting antidepressant, clorgyline, or tricyclic antidepressants (imipramine and clomipramine), did not change baseline levels of either prolactin or corticosterone. Long-term but not short-term clorgyline treatment attenuated 8-OHDPAT's effect on plasma prolactin but not on corticosterone. On the other hand, long-term but not short-term treatment with clomipramine and to some extent imipramine also, accentuated 8 OHDPAT's effect on plasma prolactin but not on corticosterone. These findings demonstrate that long-term antidepressant treatment in rats produces a differential effect on 8-OHDPAT-induced increases in plasma prolactin and corticosterone, which is consistent with other clinical and animal studies demonstrating a differential effect of long-term antidepressant treatment on two different 5-HT-mediated neuroendocrine functions. PMID- 2975225 TI - Blockade of potentiated startle responding in rats by 5-hydroxytryptamine1A receptor ligands. AB - A potentiated whole-body startle response was produced in rats by pairing intense acoustic stimuli with a light (conditional stimulus) formerly presented contiguously with electric shock. Administration of the selective serotonin (5 HT) 1A ligand 8-hydroxy-2-(di-n-propylamino)tetralin (0.125 and 0.5 mg/kg) blocked the potentiation of startle induced by the conditional stimulus. 1-[3 Chlorophenyl]piperazine, a non-anxiolytic 5-HT1B/1C agonist, did not block potentiated startle, even at a dose (1.0 mg/kg) that induced significant overall decreases in startle amplitude. The non-benzodiazepine anxiolytics buspirone (1.25-5.0 mg/kg), gepirone (3.0-10.0 mg/kg) and the related 5-HT1A ligand ipsapirone (1.0-10.0 mg/kg) blocked potentiated startle, though effects of the non-selective serotonin antagonist methysergide (0.3-10.0 mg/kg) fell short of significance. These data support a role for the 5-HT1A binding site in the anti anxiety effects of buspirone and related compounds over a range of behavioral procedures. PMID- 2975227 TI - The effect of indobufen on the thrombogenic potential of a Dacron prosthesis in an artificial circulation. AB - We have evaluated the effect of indobufen on the potential thrombogenicity of a Dacron vascular prosthesis in an artificial circulation. In a randomised double blind crossover study, ten healthy volunteers received indobufen 200 mg or placebo twice daily for one week. The artificial circulation, incorporating a 15 cm length of 8 mm Dacron graft was perfused for 60 mins with volunteer blood containing autologous 111In labelled platelets. Graft thrombogenicity was assessed by changes in platelet function, isotope labelled platelet studies and scanning electron microscopy. Platelet count fell significantly during graft perfusion (P less than 0.05) and aggregation was significantly inhibited by treatment with indobufen pre perfusion compared with the placebo group (P less than 0.02). While deposition of labelled platelets was not statistically significant, consumption of these platelets was greater in the placebo group (P less than 0.01). Field counts of adherent platelets made from scanning electron micrographs of the indobufen treated group were significantly lower (P less than 0.01) compared with the placebo group. We conclude that indobufen can reduce the thrombogenic potential of Dacron vascular grafts and suggest that it may be an effective antiplatelet agent for use following Dacron bypass surgery. PMID- 2975228 TI - Transabdominal or retroperitoneal approach to the aorto-iliac tract: a pulmonary function study. AB - An unselected consecutive series of 30 males, receiving an aorto-bifemoral Dacron graft for occlusive arterial disease, were randomised preoperatively to a transperitoneal or extraperitoneal approach. Pulmonary function tests (Forced Vital Capacity--Forced Expiratory Volume at 1 s) were performed once preoperatively and repeated four times postoperatively. As far as pulmonary function tests are concerned the results clearly demonstrated the superiority of the retroperitoneal to the transperitoneal approach. It is therefore recommended that the extraperitoneal approach should be used more frequently in reconstructive aorto-ilio-femoral surgery. PMID- 2975229 TI - [Adrenergic mechanisms regulating coronary blood outflow into the left and right sections of the heart]. AB - The alteration of veno-luminal drainage into the left heart chambers and of coronary drainage into the right heart chambers during adrenaline infusion were studied in isolated perfused at constant pressure cat hearts. During adrenaline infusion the coronary resistance was reduced, the coronary drainage into right chambers increased due to beta-adrenergic dilation mechanisms, and the luminal drainage into left chambers did not alter due to alpha-adrenergic constriction mechanisms. Thus, adrenergic regulatory mechanisms prevent an enlargement of veno luminal drainage into the arterial heart chambers. Reactions of the large coronary arteries seem to determine the alterations of luminal drainage into left chambers, whereas reactions of small coronary arteries determine alterations of coronary drainage into right chambers during the adrenaline infusion. PMID- 2975230 TI - Hepatitis B vaccine: use in dental hygiene programs. PMID- 2975232 TI - [Cobalt allergy with special reference to the hard metal industry]. PMID- 2975231 TI - Acceptance of hepatitis B vaccination by dental hygienists in the United Kingdom. PMID- 2975233 TI - [Metallic nickel on skin and in artificial sweat. Case studies]. PMID- 2975234 TI - Keratosis follicularis spinulosa decalvans with birefringent hairs. An association with variable keratoderma. PMID- 2975235 TI - Identification and immunolocalization of the laminin binding protein from embryonic avian corneal epithelial cells. AB - We report the identification of a 65-kDa laminin-binding protein (LBP) on the basal cell surface of embryonic corneal epithelium in chicken. The 65-kDa LBP was isolated by affinity chromatography with laminin-Sepharose. When reconstituted with lipid vesicles, it demonstrated specific binding for laminin. We produced monoclonal antibodies (MAbs) against 65-k Da LBP; these MAbs immunohistochemically localized to the basal epithelial cell surface. One MAb interfered with the binding of laminin to isolated epithelia and purified 65-k Da LBP. It appears that we have identified, at least in part, a cell-surface binding site for laminin. This site would provide the important link between the extracellular laminin and the intracellular cytoskeleton, and potentially the metabolic machinery of the corneal epithelial cell. PMID- 2975237 TI - [Pulmonary embolism today]. PMID- 2975236 TI - [Chrono-physiological characteristics of the enzymatic status of lymphocytes and thrombocytes in healthy children in the early postnatal ontogenesis]. PMID- 2975238 TI - [Acute coronary occlusion during transluminal angioplasty: role of coronary thrombosis]. AB - We here report on 2 patients treated by transluminal coronary angioplasty, who presented baseline angiographic aspects of an intracoronary thrombus upon vessel stenosis. In both cases mechanical dilatation was successful in increasing vessel diameter, but was complicated by activation of the thrombotic process with clot proliferation--as shown by multiple coarse filling defects irregularly stained by contrast material--and vessel occlusion. The intracoronary injection of streptokinase achieved partial slowing of the thrombotic process, but did not succeed in inhibiting it completely or in restoring vessel patency. As clinical conditions were stable, the patients were not sent to emergency surgery, but were treated conservatively with anticoagulants and platelet inhibitors: in only one patient the procedure was followed by moderate myocardial enzyme release. In both cases the coronary artery was patent at short term angiographic control. These 2 cases confirm that in the outset of transluminal angioplasty an acute coronary occlusion can be managed conservatively by thrombolytic treatment when thrombus formation can be clearly identified the cause of vessel occlusion: the activation of spontaneous lytic systems can completely restore vessel patency. If coronary occlusion was of short duration or collateral supply was adequate, myocardial infarction may not occur and emergency coronary surgery will not be necessary. PMID- 2975239 TI - [Pelviscopic treatment of abscess-forming inflammations of the pelvis]. AB - Between 1983 to 1986, in the Department of Gynaecology & Obstetrics at the University of Kiel, 50 cases with abscess-forming pelvic inflammation were treated; 28 of these through pelviscopy, whilst laparotomy was necessary in the other 22 cases. The mean age of those patients treated through pelviscopy was significantly lower than that of those subjected to laparotomy. No significant differences were found regarding either the duration of inpatient and postoperative treatment or the duration of antibiotic therapy. In the patients subjected to laparotomy, extirpation of the inflamed organs was carried out i.e. unilateral or bilateral salpingectomy, ovariectomy with eventual hysterectomy in some cases. In the pelviscopic treatment, conservative surgery was considered and ovariectomy or salpingectomy was necessary only in small number of cases. In young women, wanting to complete their families, conservative organ-retaining pelviscopic treatment of abscess-forming pelvic inflammation is a valuable alternative to laparotomy. PMID- 2975240 TI - [Quantitative hCG spot test of the serum and peritoneal fluid in intrauterine and tubal pregnancy]. AB - The concentration of hCG was measured in serum and peritoneal fluid (PF) of women with normal intrauterine pregnancy (34), extrauterine pregnancy (21), and intrauterine abortion (12) by two different methods. A new rapid, quantitative method (Spot-Test; ICON-QSR) which provides the results within 10 minutes was compared to an immunofluorometric method requiring 90 minutes (IFMA). The serum concentration of hCG and the ratio of hCG in serum and in the PF correlated well when determined by both methods (p less than 0.001). Comparable results were also obtained when hCG was measured in the serum of 4 patients with extrauterine pregnancy for a period of 2 to 3 weeks before and after surgery. When hCG was measured by means of the spot test in samples of undiluted serum containing less than 30 IU/l, lower values were obtained as compared to the IFMA. In samples containing more than 200 IU/l, the relationship was reversed. In all cases of intact intrauterine pregnancy, the level of hCG in serum exceeded that in PF, whereas the converse was observed in 19 out of 21 tubal pregnancies. These results show the spot test to be suitable for the rapid and quantitative determination of hCG in serum and the PF, provided that there is no strong hemolysis in the samples. When the concentration of hCG exceeds 300 IU/l, the measurement has to be repeated with diluted samples. Differences in concentrations of less than 20% cannot always be distinguished. Even though the results of the present study are promising, the usefulness of the spot test for routine clinical practise has to be examined in further studies. PMID- 2975241 TI - [Therapy with gonadotropin releasing hormone agonists (zoladex) in premenopausal females with metastastic breast cancer]. AB - Medicinal castration using GnRH-analogues is a new therapeutic possibility for treating metastasizing breast cancer in premenopausal women. A total of 22 premenopausal patients were included in the study reported here, all of them low risk cases. Twenty of the 22 patients had hormone receptor-positive primary tumors. A slow-release depot form of Zoladex (ICI 118630) was used as a GnRH agonist and was administered subcutaneously (3.6 mg) at four-week intervals. The long-term administration of Zoladex brought about a significant reduction in blood FSH, estradiol, and progesterone levels within one to four weeks. In contrast, there were no detectable changes in ACTH, DHEAS, cortisol, testosterone, prolactin, or androstendion levels. Therapy-induced amenorrhea occurred in all cases. The objective remission rate achieved (complete and partial remission) was 45%. As opposed to other formulations, the use of Zoladex as a GnRH analog in depot form has significant advantages, which become particularly evident through improved compliance. With Zoladex therapy an effective drug-induced castration can be accomplished in premenopausal women. As regards its efficacy it is comparable to an ovarectomy, though with less pronounced side effects. PMID- 2975242 TI - [Immunosuppressive properties of a Soviet preparation of cyclosporins]. PMID- 2975243 TI - [A rare genotype of the rhesus system (--D--/--D--)]. PMID- 2975245 TI - Vascular vasopressin receptors. AB - 1. Vascular vasopressin receptors are understood because of the specific application of each major technical advance in pharmacology; this review shows that isolated organs, whole animal preparations, hormone synthesis, radioligand binding, and human studies have all played their part. 2. Even so, neither vascular vasopressin receptor heterogeneity nor occupancy-response relationships are fully understood; by way of comparison far more is known about alpha adrenoceptors. 3. The biochemical pharmacology of vascular vasopressin receptor activation is still in its infancy. Whilst the second messenger molecules resulting from vascular vasopressin receptor activation appear to be component(s) of the pathways of phosphoinositide metabolism, technical difficulties have led investigators to study similar vasopressin receptors in other tissues. 4. It is not certain, for example, that results from hepatic vasopressin receptor studies can be automatically extrapolated to vascular smooth muscle. 5. Lastly, the directions that vascular vasopressin research might take are speculated on. It is not known whether the vascular vasopressin receptor is itself a polymer, whether receptor heterogeneity could be exploited in the clinical uses of vasopressins, nor whether vasopressins are co-released with other neurotransmitters. PMID- 2975244 TI - [A case of combined carriage of HbE and beta-thalassemia in the Georgian SSR]. PMID- 2975246 TI - Exercises for prevention and alleviation of back pain. PMID- 2975247 TI - The frequency and profile of medically compromising conditions in an dental school patient population. PMID- 2975248 TI - Aspergillus nidulans contains a single actin gene which has unique intron locations and encodes a gamma-actin. AB - The single actin gene from the filamentous fungus Aspergillus nidulans has been isolated and characterized. The only other organism reported to contain just one actin gene is another Ascomycete, the budding yeast Saccharomyces. The nucleotide sequence of the A. nidulans actin gene predicts a polypeptide containing the N terminal sequence identifying the gamma-actin isotype. Until now this characteristic N terminus has only been reported to occur in vertebrate actin sequences. A monospecific anti-gamma-actin antiserum recognizes a single 42-kDa band in immunoblots of total Aspergillus protein. None of the six introns in the A. nidulans actin gene sequence aligns precisely with those found in other actin genes. One, unlike other known actin introns, is located in the 3'-untranslated region of the gene. The 5' and 3' ends of the gene have been characterized. The Aspergillus actin gene has a heterogeneous transcript size due to the presence of several different 3' termini. Of four characterized polyadenylated transcripts, only the longest contains a typical AATAAA polyadenylation signal near its 3' terminus. Using an integrative plasmid containing Aspergillus actin sequences and the pyr4 gene from Neurospora, the A. nidulans actin gene has been mapped to the first chromosome. PMID- 2975250 TI - Escherichia coli sbcC mutants permit stable propagation of DNA replicons containing a long palindrome. AB - Recombinant DNA libraries generated in vitro should in theory contain all of the sequences of the genomes from which they are derived. However, the literature is dotted with reports of sequences that cannot be recovered, are under-represented, or are highly unstable. In particular, long palindromic nucleotide sequences of perfect or near-perfect symmetry are either lethal to the vector or suffer deletions or other rearrangements that remove symmetry [Collins, Cold Spring Harbor Symp. Quant. Biol. 45 (1981) 409-416; Collins et al., Gene 19 (1982) 139 146; Hagan and Warren, Gene 24 (1983) 317-326]. We report here that mutation of a single gene, namely sbcC, can overcome this inviability and allow for the stable propagation of a 571-bp nearly perfect palindrome in Escherichia coli. This has implications for the choice of strains used for the recovery and analysis of cloned nucleotide sequences. PMID- 2975249 TI - Analysis of the Escherichia coli glycogen gene cluster suggests that catabolic enzymes are encoded among the biosynthetic genes. AB - The nucleotide sequences of the Escherichia coli genome between the glycogen biosynthetic genes glgB and glgC, and 1170 bp of DNA which follows glgA have been determined. The region between glgB and glgC contains an open reading frame (ORF) of 1521 bp which we call glgX. This ORF is capable of coding for an Mr 56,684 protein. The deduced amino acid (aa) sequence for the putative product shows significant similarity to the E. coli glycogen branching enzyme, and to several different glucan hydrolases and transferases. The regions of sequence similarity include residues which have been reported to be involved in substrate binding and catalysis by taka-amylase. This suggests that the proposed product may catalyze hydrolysis or glycosyl-transferase reactions. The cloned region which follows glgA contains an incomplete ORF (1149 bp), glgY, which appears to encode 383 aa of the N terminus of glycogen phosphorylase, based upon sequence similarity with the enzyme from rabbit muscle (47% identical aa residues) and with maltodextrin phosphorylase from E. coli (37% identical aa residues). Results suggest that neither ORF is required for glycogen biosynthesis. The localization of glycogen biosynthetic and degradative genes together in a cluster may facilitate the regulation of these systems in vivo. PMID- 2975251 TI - [Relation between health status indicators and physical fitness of students and their physical activity]. PMID- 2975252 TI - [A method of determining the leading factors in professional orientation of adolescents at the inter-school educational-industrial combine]. PMID- 2975253 TI - [Radiation-hygienic principles of optimal radiation monitoring in the surveillance areas of atomic power stations]. PMID- 2975254 TI - [Incidence of acute respiratory infections among children exposed to atmospheric pollution]. PMID- 2975255 TI - [Significance of socio-hygienic conditions at food-processing plants for securing the quality of food products]. PMID- 2975256 TI - [Changes in the activity of a number of enzymes of skeletal muscle fibers in rats after a single physical load]. PMID- 2975257 TI - [Effectiveness of measures with regard to rationalization and improvement of working conditions of flour-mill workers]. PMID- 2975258 TI - [Indicators of physical development of younger schoolchildren in Nagorny Karabakh]. PMID- 2975259 TI - [Blood flow in the umbilical vein measured by the ultrasonographic method using the Doppler attachment (preliminary report)]. PMID- 2975260 TI - Infectious disease update. PMID- 2975261 TI - [Surgical treatment using microdissection of compressive lumbar pain syndrome caused by a herniated disk and degenerative diseases]. PMID- 2975262 TI - Common acute lymphoplastic leukemia antigen (CALLA) and intestinal metaplasia. AB - Intestinal metaplasia (IM) foci in 19 antral and 14 fundal gastric biopsies from patients with chronic atrophic gastritis were studied immunohistochemically for the presence of CALLA antigen. In only 2 cases were metaplastic glands completely negative, in 14 cases they were all positive, and in 17 cases variable proportions of CALLA positive and negative metaplastic glands were present. Complete IM seems to be less frequent than the incomplete type when the presence of CALLA is taken in consideration. CALLA is obviously a much better marker for complete or incomplete small IM. The possible importance of the presence of CALLA in IM foci for the future development of gastric cancer is discussed. PMID- 2975263 TI - Regulation of pathways of glucose metabolism in the kidney. The activity of the pentose phosphate pathway, glycolytic route and the regulation of phosphofructokinase in the kidney of lean and genetically obese (ob/ob) mice; comparison with effects of diabetes. AB - The activities of enzymes of the glycolytic route, the pentose phosphate pathway and NADPH-linked enzymes have been measured in the kidneys of genetically obese (ob/ob) mice and their lean litter mates. The renal content of glucose 6 phosphate (G6P), fructose 6-phosphate (F6P), fructose 1,6-bisphosphate (Fru-1,6 P2) and fructose 2,6-bisphosphate (Fru-2,6-P2) were also measured. Increases were found in hexokinase and enolase with an upward trend in pyruvate kinase in the ob/ob mouse kidney; a significant decline in malic enzyme was also seen. The renal content of G6P and Fru-1,6-P2 increased. There was no renal hypertrophy despite a degree of hyperglycaemia, which was, however, considerably below that observed in experimental diabetes. Comparison of the renal changes in the hyperglycaemic-hyperinsulinaemic ob/ob mice with the hyperglycaemic hypoinsulinaemic diabetic group showed two distinct groupings. Firstly, changes which were similar in the two groups included: increases in hexokinase, G6P and Fru-1,6-P2, and a decrease in malic enzyme. Secondly, opposite changes were seen in enolase and in enzymes at the G6P crossroads, phosphoglucose isomerase and phosphoglucomutase. The elevated hexokinase and G6P in both ob/ob and diabetic groups may be involved in the eventual accumulation of basement membrane material in the glomerulus which is a common feature of the two conditions. PMID- 2975265 TI - Human atrial natriuretic polypeptide in plasma of patients with anorexia nervosa. AB - To examine the effects of chronic dehydration and starvation on plasma levels of human atrial natriuretic polypeptide (hANP) in human subjects, the basal level and saline-induced rise of plasma hANP in 7 patients with anorexia nervosa were compared with those in age-matched healthy subjects. The unstimulated level of plasma hANP was markedly high in the patients with anorexia nervosa (patients vs. control; 55.4 +/- 9.0 pg/ml vs. 11.4 +/- 6.1 pg/ml, P less than 0.01). However, no significant increase of plasma hANP in the anorectic patients was observed in response to saline-infusion, while a 3-fold increase over the basal level of plasma hANP was noted in the saline-infused normal young subjects. These results show that hANP may be secreted to an inadequate extent, hence the release would be resistant to volume-loading. The pathophysiological meaning of such a high plasma concentrations of hANP in anorexia nervosa is the subject of ongoing studies. PMID- 2975264 TI - The effect of beta-endorphin on basal and TRH-stimulated TSH release in conscious male rats. AB - The inhibitory effect of beta-endorphin (EP) or other opioids on TSH secretion is, in contrast to their stimulating properties on PRL release, still a matter of debate. In the present study a dose of 1 microgram beta-EP injected intracerebroventricularly (IVT) in unstressed conscious male rats, though highly effective on PRL release, did not affect basal TSH levels, nor the TRH-induced TSH secretion. The previously reported inhibition of TSH release by opioids may therefore be an effect only seen when pharmacological doses are used. PMID- 2975266 TI - Dental implications of pharmacological management of the Alzheimer's patient. PMID- 2975267 TI - Professional dental care for patients with dementia. PMID- 2975268 TI - Up-to-the-minute news on AIDS. PMID- 2975269 TI - [Josamycin in the treatment of acne vulgaris. Results of a multicenter study of 745 patients]. PMID- 2975270 TI - [Action of oral mesoglycan in dermatology]. PMID- 2975271 TI - Enhancement of allo-responsiveness of human lymphocytes by acemannan (Carrisyn). AB - Healing powers have been imputed as being a feature of the gel from the aloe vera plant for centuries. The recent isolation of the active ingredient, acemannan, has made testing of this drug important. Since the drug appears to enhance monocyte function in other experiments, these studies were designed to test the capacity of acemannan to enhance immune response to alloantigen and to test whether the potential enhancement is a monocyte driven phenomenon. Acemannan did not enhance lymphocyte response to syngeneic antigens in the mixed lymphocyte culture (MLC) but importantly increased alloantigenic response in a dose-response fashion (2.6 x 10(-7) - 2.6 x 10(-9)M). This effect of acemannan was shown to be a specific response and to concur with concentrations of in vitro acemannan achievable in vivo. A separate series of mixing experiments demonstrated that acemannan incubation with monocytes permitted monocyte driven signals to enhance T-cell response to lectin. It is concluded that acemannan, the active ingredient of the aloe vera plant, is an important immunoenhancer in that it increases lymphocyte response to alloantigen. It is suggested that the mechanism involves enhancement of monocyte release of IL-I under the aegis of alloantigen. This mechanism may explain in part the recently observed capacity of acemannan to abrogate viral infections in animal and man. PMID- 2975272 TI - Role of Doppler ultrasound flow studies in the diagnosis of temporal arteritis. PMID- 2975274 TI - Redesigning the workplace prevents low-back injuries. PMID- 2975273 TI - [Emergency bypass operation following unsuccessful percutaneous transluminal coronary angioplasty: how important is ischemia time in preventing an infarct?]. PMID- 2975276 TI - Dental care for AIDS patients. PMID- 2975275 TI - Purification and characterization of a sperm motility inhibitor in human seminal plasma. AB - A sperm motility inhibitor from human seminal plasma was purified and characterized. The purification procedure includes dialysis, ion exchange chromatography on SP-Sephadex C-25 and adsorption chromatography on hydroxylapatite. With this procedure, the seminal plasma motility inhibitor was purified 290-fold with a 24% recovery in inhibitory activity. Its molecular weight has been estimated at 18,000 to 22,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, but at 13,000 to 15,000 according to molecular sieving under native conditions. The mobility inhibitor has an isoelectric point pH 9.1. It is stable over a wide range of pH (5 to 10) and at temperatures up to 60 C. The observation that the seminal plasma factor inhibited purified bull dynein ATPase in a concentration-dependent manner may suggest that it blocks the motility of demembranated spermatozoa by interfering with dynein arm function. PMID- 2975277 TI - Characteristics and functional significance of canine abdominal muscles. AB - To assess the characteristics and function of the muscles of the anterolateral abdominal wall, we have examined the isometric contractile properties of bundles of canine rectus abdominis (RA) and external oblique (EO) muscles. In addition, we have related the lengths of these muscles measured sonometrically in vivo at supine functional residual capacity (FRC) to in vitro optimal force-producing length (Lo). We also investigated the action of the abdominal muscles on the displacement of costal and crural diaphragm. We found that 1) contraction time of RA was longer and that the RA developed greater force than the EO at submaximal stimulation frequencies; 2) maximal tetanic force and the active length-tension curves were similar in both abdominal muscles; 3) on passive stretch, the compliance of the RA was one-third that of the EO; 4) at supine FRC, the EO is operating at 83% of Lo, whereas the RA is operating at 105% of Lo; 5) stimulation of either RA or EO (abdominal pressure of 15 cmH2O) lengthened the costal and crural diaphragm toward their Lo values, with greater crural excursion occurring than costal. We conclude that the RA is well suited for restraining the abdominal viscera in prone quadrupeds, whereas the EO is better designed to assist expiration. Stimulation of both muscles improves in situ diaphragmatic operating length. PMID- 2975278 TI - Pulmonary hypertension and pulmonary vascular reactivity in beagles at high altitude. AB - It is unclear whether dogs develop pulmonary hypertension (PH) at high altitude. Beagles from sea level were exposed to an altitude of 3,100 m (PB 525 Torr) for 12-19 mo and compared with age-matched controls remaining at low altitude of 130 m (PB 750 Torr). In beagles taken to high altitude as adults, pulmonary arterial pressures (PAP) at 3,100 m were 21.6 +/- 2.6 vs. 13.2 +/- 1.2 Torr in controls. Likewise, in beagles taken to 3,100 m as puppies 2.5 mo old, PAP was 23.2 +/- 2.1 vs. 13.8 +/- 0.4 Torr in controls. This PH reflected a doubling of pulmonary vascular resistance and showed no progression with time at altitude. Pulmonary vascular reactivity to acute hypoxia was also enhanced at 3,100 m. Inhibition of prostaglandin synthesis did not attenuate the PH or the enhanced reactivity. Once established, the PH was only partially reversed by acute relief of chronic hypoxia, but reversal was virtually complete after return to low altitude. Hence, beagles do develop PH at 3,100 m of a severity comparable to that observed in humans at the same or even higher altitudes. PMID- 2975279 TI - Adenocarcinoma of lung and sigmoid colon. PMID- 2975280 TI - The Logan H. Roots hospitals: "a tale of two cities". PMID- 2975282 TI - The coming quality crisis. PMID- 2975281 TI - AIDS in Arkansas. AIDS 1988. PMID- 2975283 TI - Quality of care review by the PRO. PMID- 2975284 TI - Consensus conference on colorectal carcinoma. PMID- 2975285 TI - Ipratropium bromide: bronchodilator action and effect on methacholine-induced bronchoconstriction. AB - The effects of ipratropium bromide (80 and 200 micrograms) and placebo on the basal bronchial tone and on methacholine-induced bronchoconstriction were investigated in 10 asthmatic patients in a placebo-controlled double-blind manner. Bronchial hyperreactivity to methacholine was confirmed at a pretrial bronchial challenge. The patients were randomly allocated to two groups in which the drug was inhaled from either metered-dose inhalers (MDI) or powder capsules. With the high dosage, the bronchodilation resulting from powder capsules was somewhat more pronounced than that achieved with the MDI. Otherwise the bronchodilator effect of ipratropium bromide and the protection afforded by the drug against methacholine-induced bronchoconstriction were similar in the two groups. In five patients the bronchodilator effect was better and in four patients the tolerance to methacholine was greater after the higher ipratropium dosage than after the lower one. In two patients ipratropium bromide had no bronchodilator effect but gave good protection against methacholine-induced bronchoconstriction. It is concluded that some patients benefit from a dosage of ipratropium bromide higher than that usually recommended and that an anticholinergic effect on the bronchi is possible even in the absence of the bronchodilator effect in the basal state. PMID- 2975286 TI - Chronobiology and asthma. II. Body-time-dependent differences in the kinetics and effects of bronchodilator medications. AB - Several bronchodilator medications exhibit body-time (i.e., biological rhythm) dependent changes in their pharmacokinetics and effects. Epinephrine (Adrenalin), metaproterenol (orciprenaline), aminophylline, and ipratropium bromide all have a better effect on the tone of the airways during the night and/or morning, when bronchial patency is low, than during the day, when it is high. The pharmacokinetics of sustained-release theophyllines (SRTs) exhibit administration time differences. Day-night dosing-time differences in the kinetics of theophylline are especially prominent in children. Generally, in day-active asthmatic children the absorption of SRT is more rapid after a morning than an evening dosing. The administration-time effect on the kinetics of SRTs also is apparent in adult patients, but the magnitude of difference between the day versus evening administrations apparently is more moderate. Initial findings from studies of unequal (morning versus evening) BID dosing schedules--more theophylline or terbutaline before bed-time than arising--reveal a better therapeutic advantage relative to equal BID dosing schedules for those patients with predominantly nocturnal symptoms. Once-daily (OD) SRTs intended for delivery of the entire daily dose at a single time also differ quantitatively in their chronokinetics. Since asthma is mainly a nocturnal disease in many patients, it has been recommended by many that ODSRTs be taken in the evening. If taken in the morning, as is the current practice in the United States, they may not ensure therapeutic theophylline blood levels during the night when most needed. Moreover, not all ODSRTs appear suitable for once-nightly administration because of unacceptable kinetics. PMID- 2975288 TI - Six weeks to twenty-one years old: a longitudinal study of children with Down's syndrome and their families. Third Jack Tizard memorial lecture. AB - A cohort of children with Down's syndrome, and their families, have been seen at intervals from six weeks old. Some follow-up data to 21 yrs are reported here. Until 21 yrs when one mother withdrew her son there have been no losses to the study apart from those caused by death. Mean IQs rose slightly from 11 to 21; scores on language and academic tests favoured the home-reared, females, and middle-class young people even when IQ was allowed for. Some of the effects on the family of having a handicapped member were explored; although adverse effects are fewer than might have been expected many parents, especially mothers, carry significant burdens. PMID- 2975287 TI - Who remains celibate? PMID- 2975289 TI - Personality characteristics of work-ready workers' compensation clients. AB - Result of this investigation indicate that "work-ready" status constitutes a significant variable in the characterization of workers' compensation claimants. The mean MMPI profile obtained for work-ready claimants (N = 69) showed no scales elevated above 70. Previous compensation studies, which did not differentiate claimants on this basis, consistently reported an elevated (123) profile for this population. In addition to demonstrating the absence of significant psychological disturbance in this population on the MMPI, the present study also found that work-ready claimants do not differ from the general working population on a number of salient attitudinal variables (e.g., ambition, willingness to accept challenges, etc.). PMID- 2975290 TI - Acute effects of the new angiotensin-converting enzyme inhibitor cilazapril: a pilot study. AB - This study assesses the magnitude and duration of action of three different oral doses of the new orally active angiotensin-converting enzyme (ACE) inhibitor RO 312848 (cilazapril, Hoffman-LaRoche, Nutley, NJ) on blood pressure and plasma ACE levels. Twelve hypertensive patients were separated into two groups: Group A (n = 6) received two single daily doses of 5 and 10 mg, each preceded and followed by two placebo days, and Group B (n = 6) received 10 and 20 mg on an identical protocol. The onset and duration of an appreciable blood pressure lowering effects were at 2 hours and at least for 12 hours, respectively, whereas suppression of ACE levels occurred at 1 hour and lasted for more than 72 hours. Response of these two parameters was partial after 5 mg, but was maximal after 10 mg and did not increase further with the 20-mg dose. A 5-10 mg dose daily may be sufficient to maintain chronic blood pressure control with this agent, but long term dose trials are necessary to establish its clinical utility. PMID- 2975291 TI - An acute dose-response pharmacodynamic evaluation of orally administered isradipine (PN-200-110) in hypertensive patients. AB - This study was conducted to assess acute oral dose-responses of four dose levels (2.5, 5, 10, 20 mg) of isradipine, a new calcium channel blocking agent of the 1,4-dihydropyridine group. Sixteen patients with mild essential hypertension were investigated using a randomized, four-way cross-over, double-blind, placebo controlled design. After a 2-week washout period, all patients were admitted to a research unit where they entered a 3-day placebo equilibration, followed by 9 days of double-blind single doses of isradipine. Drug administration was randomized, and baseline blood pressure values obtained preceding active medication doses were fairly constant. Dose-related reductions in supine and in standing blood pressures were obtained. The mean peak supine blood pressure decrements to doses of 2.5, 5, 10, and 20 mg were 17/16, 25/19, 35/22, and 37/25 mm Hg, respectively. Pulse rates increased slightly. Similar responses were obtained with patients in the erect position. Peak hypotensive responses occurred within 3 hours after dosing. The duration of effect persisted as long as 21 hours, particularly after the 10- and 20-mg doses (15/9 and 17/12 mm Hg mean supine blood pressure decrements, respectively). PMID- 2975292 TI - Childhood affective disorders. PMID- 2975294 TI - Manic symptoms in a non-referred adolescent population. AB - Using a structured interview with 150 randomly selected 14-16-year-olds and their parents, we found 20 who endorsed four or more manic symptoms of at least 2 days duration. Compared to the rest of the sample, these teenagers had significantly higher rates of attention deficit, conduct, and anxiety disorders and psychotic symptoms, and were seen as needing treatment by the interviewers. However, most of the pathology was endorsed by teenagers rather than parents. Other assessments confirmed this group as dysphoric, impulsive and emotionally labile. We discuss the normative and diagnostic implications of these findings though long-term follow-up is necessary to draw more certain conclusions. PMID- 2975293 TI - Depressive disorders in childhood. III. A longitudinal study of comorbidity with and risk for conduct disorders. AB - As part of a longitudinal nosologic study of major depressive disorder (MDD), dysthymic disorder, and adjustment disorder with depressed mood in 104 school aged probands, the prevalence and consequences of comorbid conduct disorders (CD) were examined. During the index depressive episodes, 16% of the patients had comorbid CD; during the full study observation 23% had CD; and the estimated time dependent risk of conduct disorder developing was 36% by age 19. For most cases, comorbid CD developed as a complication of the depression and persisted after the depression remitted. Comorbid CD was not differentially associated with the type of depression at study entry, did not affect depressive symptom presentation, was similarly distributed among boys and girls, and was unrelated to demographic factors. Additionally, comorbid CD did not affect recovery from the index depressive episodes and did not influence the symptom-free interval before a recurrent depression among cases with MDD. The risk of CD developing was not altered by chronologically earlier family variables or demographic factors. But girls who had attention deficit disorder, compared to those who did not, seemed to be at higher risk for CD during study observation. Finally, in this depressed cohort, having CD any time was associated with an increased rate of long-term functional problems. PMID- 2975295 TI - Comorbidity of mental disorders in the post-mortem diagnosis of completed suicide in children and adolescents. AB - In a psychological autopsy of 21 children and adolescents aged 11-19 years who committed suicide as compared with a matched-pair control group, 95% of the suicide victims and 48% of the controls had at least one serious diagnosable mental disorder (P less than 0.02). The presence of two or more mental disorders (comorbidity) was the rule occurring in 81% of the suicide victims and 29% of the controls (P less than 0.001). Mood disorders such as major depression, major depression superimposed on dysthymic disorder and dysthymic disorder coexisted with either alcohol and drug abuse, conduct disorder or other mental disorders in 76% of the victims and 24% of the controls (P less than 0.025). The suicide victims experienced significantly more psychosocial stressors and the poorest level of adaptive functioning. This study suggests that suicide in children and adolescents very frequently is the outcome of serious psychiatric disorders, particularly when associated with drug and alcohol abuse. PMID- 2975296 TI - Course of major depression in non-referred adolescents: a retrospective study. AB - This article reports on a naturalistic study of the course of illness of 38 children diagnosed as having a current or past episode of major depression out of a sample of 275 children who were selected by a method not related to their psychopathology or treatment-seeking behavior. Assessments of the presence of depression and the course of this disorder were made using structured clinical interviews (DICA and DICA-P) and a criterion-based diagnostic system (DSM-III). Longitudinal methods of data analysis included the use of life tables. The proportion of children depressed for 2 years closely resembles the results found in investigations of children who sought treatment for a psychiatric disorder. The probability of remaining depressed in these children was 21% at 1 year after onset, and 10% at the 2-year point. This parallels the rate of chronicity and the decline in rates of recovery which occur over time in adult depression. PMID- 2975297 TI - Childhood-onset depressive disorders. A follow-up study of rates of rehospitalization and out-of-home placement among child psychiatric inpatients. AB - This report describes preliminary outcome data for a sample of child psychiatric inpatients with diagnoses of major depression and/or dysthymic disorder at the time of their hospitalizations. Depressed children were compared with a contrast group of children with schizophrenia spectrum disorders. Results (based on semi structured telephone interviews) indicate high rates of rehospitalization among our depressed cohort. Depressed children had rehospitalization rates of 35% and 45% respectively in the first and second years after discharge. Out-of-home placement was rarer in the depressed group, and significantly less likely than for children with schizophrenia spectrum disorders. However, 15% of the depressed cohort were placed out of their homes within the first year of discharge. There were no differences between children with major depressive and dysthymic disorders on these outcome variables, underscoring the serious long-term correlates of childhood dysthymic as well as major depressive disorders. PMID- 2975298 TI - A family study of bipolar I disorder in adolescence. Early onset of symptoms linked to increased familial loading and lithium resistance. AB - Lifetime rates of psychiatric illness were compared in relatives of adolescent probands with bipolar I disorder and in relatives of age-matched schizophrenic controls. Familial aggregation of major affective disorders was observed in bipolar probands, the rate of bipolar I disorder greatly exceeding that reported in relatives of adult bipolar probands. Adolescent probands with childhood onset of psychiatric disturbance were distinguished from probands who had no premorbid childhood psychiatric abnormality in two ways: (1) significantly increased aggregation of bipolar I disorder in first-degree relatives; and (2) poorer antimanic response to lithium carbonate. These data underscore important heterogeneity in adolescent-onset bipolar disorder. PMID- 2975299 TI - Early-onset major depression in parents and their children. AB - In a study of 6-23-year-old offspring of depressed and of normal parents, an inverse relationship between the rates of major depression among the children and the age of onset of major depression in their proband parents was found. The children of parents who had an onset of major depression that was younger than age 20 years overall had the highest risk of major depression. There was specificity in the findings in that these higher rates were nearly all accounted for by prepubertal onsets of major depression in their children. There was a 14 fold increased risk of onset of depression before age 13 in the children of probands who had onset less than age 20. These results were not confounded by the current age of the proband or the children, by interview status (children were interviewed), by comorbidity in the parents or by assortative mating. Future family genetic studies should examine the rates and patterns of illness of the biological relatives of probands with prepubertal-onset major depression. PMID- 2975300 TI - Parental concordance for affective disorders: psychopathology in offspring. AB - This paper examines the effects of parental concordance for affective disorders and psychopathology among the 219 offspring of probands with major depression and normal controls. The lifetime prevalence of psychiatric disorders was significantly higher among the spouses of depressed probands as compared to those of normal controls. The spouses of 37% of the normals and 69% of the depressed probands met criteria for a diagnosis of major depression, an anxiety disorder, or alcoholism. Parental concordance for diagnoses, particularly for anxiety disorders, substantially increased the risk of major depression and anxiety disorders in their children. Moreover, the marital relationship, some aspects of family adjustment and severity of current symptoms were significantly worse among the couples who exhibited diagnostic concordance for anxiety, alcoholism and/or depression. The major implication of these findings is that the diagnostic status of both parents should be considered in the design and analysis of studies of children. The findings of the present study also underscore the importance of assessment of comorbid disorders in parents and offspring. Although the original study design focused on the risk of depression in children of parents in treatment for major depression, stronger transmissibility was found for anxiety disorders plus depression than for major depression alone. However, the exclusion criteria of a lifetime history of mania or hypomania led to an extremely low proportion of probands with pure major depression without concomitant anxiety disorders. These findings confirm the results of previous studies which have demonstrated a strong degree of overlap between affective and anxiety syndromes. The increased risk of anxiety disorders in the offspring of parents who had sought treatment for non-bipolar major depression suggests that anxiety may constitute an early form of expression of affective disorders. Confirmation of the finding of age-dependent expression of anxiety and depression in prospective longitudinal studies of children is indicated. PMID- 2975301 TI - School-aged children of depressed parents: a blind and controlled study. AB - This study examines the prevalence of psychiatric dysfunction in the children of parents diagnosed with affective disorders. Sixty children from 37 proband families were compared to 43 children from 26 families obtained from matched controls as well as 20 children from 13 medically ill families. Group differences in diagnosable childhood disorders and familial characteristics are investigated. Significantly more disorders and symptoms were noted in the children with psychiatrically ill parents as compared to children from matched controls and medically ill parents. Using logistic and Cox survival analyses, correlates for the risk of affective disorder, attention deficit and conduct disorder in the children were examined. Maternal depression and paternal alcoholism were related to the risk for depression in the child. The child's sex and the presence of affective disorders in the father were significantly related to the risk for attention deficit disorder. Maternal alcoholism, parental divorce and the type of subject (proband or control family) were significantly related to the risk for conduct disorder. The findings are discussed relative to results from earlier studies on rates of disorder in the offspring of depressed parents. PMID- 2975302 TI - Affective disorder in childhood: separating the familial component of risk from individual characteristics of children. AB - In studying the risk of affective disorder in children, the investigator must deal with the problem that there are two possible units of analysis: the child and the family. An analysis based on children must take account of the intercorrelation within a sibship to produce correct results, while a family based analysis makes it difficult to investigate individual characteristics of children that help determine the net risk. A two-stage iterative approach to this problem is proposed, yielding estimates of the effect of family-based factors (parental illness, family social class, marital status of parents) and individual factors (age and sex of child, previous non-affective illness). This technique is applied to a sample of 275 children from 143 families representing a wide range of familial risk for affective disorder. The final family-based model (predicting at least one child with affective disorder in the sibship) indicates a six-fold increase in risk to the child associated with maternal affective disorder (P less than 0.001), a three-fold increase in risk associated with paternal affective disorder (P less than 0.05) and divorce or separation of the biological parents, and a suggestion of increased risk in the highest social class (P = 0.06). The excess sibship risk, due to child factors age, prior anxiety disorder, and prior childhood diagnosis, contributed significantly to the family prediction (P less than 0.001). PMID- 2975303 TI - Psychiatric disorder in adolescent offspring of parents with affective disorder in a non-referred sample. AB - The relationship between parental psychopathology and psychiatric disturbance in 153 offspring aged 6-19 was assessed in 81 families randomly selected from a prepaid health plan. Offspring of parents with a history of affective disorders and of parents with non-affective psychiatric disorders had higher rates of psychiatric diagnoses and poorer adaptive functioning than children of parents who had never experienced a psychiatric illness. Offspring whose parents had affective disorder had a rate of affective disorder of 30% compared to a rate of 2% in the rest of the sample. This relationship between parental affective disorder and poor child outcome was observed when the separated and divorced families were removed from the analyses. PMID- 2975304 TI - Growth hormone response to desmethylimipramine in depressed and suicidal adolescents. AB - Desipramine 75 mg i.m. was given in the morning to 20 adolescents with major depressive disorder and 23 normal controls. Depressed adolescents secreted significantly less growth hormone (GH) over the next 2 h than did normal adolescents, although a substantial proportion of the differences were accounted for by the depressed adolescents who had a specific suicidal plan or attempt during the episode. Severity of depression or the presence of other depressive symptoms did not predict GH secretion within the depressed group. Age, sex and maturational factors in the control of GH are discussed. It is concluded that these differences in GH secretion probably reflect differences in CNS beta adrenergic and/or serotonergic function. Suicidality and depression may have different psychobiological correlates in adolescents. PMID- 2975305 TI - Self-report vs. biological markers in assessment of childhood depression. AB - Sensitivity, specificity, and diagnostic confidence of the Children's Depression Inventory (CDI) and the dexamethasone suppression test (DST) in assessing childhood depression were determined. Parent and child forms of the CDI and plasma cortisol levels at 8 a.m. and 4 p.m. were used. Three groups of prepubertal children were sampled: (1) 63 depressed inpatients, (2) 14 non depressed inpatient psychiatric controls, and (3) 21 normal community sample controls. Results suggest that scores greater than or equal to 15 on the CDI obtained from either parent or child provide a good screening instrument (sensitivity = 89%). If the DST is administered to all children with elevated CDI scores, diagnostic confidence is 97.5%. Cautions and recommendations for use of the CDI and DST are made. PMID- 2975306 TI - Atrial natriuretic peptide in the sheep. AB - To study atrial natriuretic peptide (ANP) physiology in the chronically catheterized pregnant sheep model we developed a heterologous radioimmunoassay for ovine ANP using an antiserum raised against 1-28 human ANP. This antiserum (Tor I) is specific for the aminoterminus of the human ANP molecule and shows little cross reaction with any carboxyterminus ANP fragments. Ovine ANP immunoreactivity was characterized using this antiserum and a commercially available carboxyterminus ANP antiserum obtained from Peninsula Laboratories. Each antiserum detected 2 peaks of immunoreactivity in ovine atrial extracts chromatographed on a Biogel P-10 column. The minor peak migrated at a position close to 125I-human ANP whereas the major peak represented a larger molecular weight species of ANP. Examination of gel filtration eluates of ovine plasma extracts showed one immunoreactive ANP peak using the Tor I assay system and 2 peaks with the Peninsula Laboratories assay. Plasma immunoreactive ANP levels were determined in 9 sheep using both radioimmunoassay systems. Mean (+/- SEM) levels were similar using the Peninsula Laboratories and the Tor I assay systems (57 +/- 8 pg/ml versus 43 +/- 4 pg/ml, P greater than 0.05). Using the Tor I antiserum, fetal plasma immunoreactive ANP levels were found to be significantly higher than maternal levels (188 +/- 17 versus 48 +/- 8 pg/ml, P less than 0.01) whereas pregnant and nonpregnant adult sheep had similar plasma immunoreactive ANP levels (48 +/- 8 versus 43 +/- 4 pg/ml, P greater than 0.05). Disappearance curves of synthetic human ANP from the plasma of maternal and fetal sheep were assessed using both immunoassay systems and found to be similar. PMID- 2975307 TI - [Alterations in arachidonic acid metabolism in aorta and gingiva of streptozotocin-induced diabetic rats]. PMID- 2975308 TI - What's right with nursing? PMID- 2975309 TI - "Infections related to intracranial pressure monitoring". PMID- 2975311 TI - Sexuality and multiple sclerosis. AB - The individual with multiple sclerosis (MS) experiences a severe alteration in lifestyle both physiologically and emotionally. Sexuality is an integral part of life. However, discussion of alterations in sexuality by health care workers has traditionally been viewed as taboo. Specifically, the sexual concerns in chronic debilitating disorders such as MS have been neglected. This paper will explore physical and psychological aspects of MS on a patient's sexuality. Implications for nursing practice will also be considered. PMID- 2975310 TI - Interpretation of ICP pulse waves to determine intracerebral compliance. AB - Recently, several authors have identified the need for monitoring cerebral compliance in addition to intracranial pressure (ICP) when managing patients with neurologic dysfunction. Analysis of the ICP pulse wave is one method for determining compliance. The ICP pulse wave, observed on a continuous real time pressure monitor display corresponding to each heart beat has three components analogous to the components of the arterial pulse wave. Analysis allows determination of intracerebral compliance. Implications for nursing care and suggestions for research based on ICP pulse wave analysis will also be discussed. PMID- 2975312 TI - Adrenal medullary transplant as a treatment for Parkinson's disease: perioperative considerations. AB - Autologous adrenal medullary transplantation, a new treatment for severe Parkinson's disease, is both complex and risky. Detailed planning is essential prior to beginning the procedure, and perioperative concerns unique to the patient and the procedure are challenging. This article reviews rationale for treatment and focuses on preoperative planning and care in addition to intraoperative concerns. PMID- 2975313 TI - Electrocardiographic changes following subarachnoid hemorrhage. AB - Electrocardiographic abnormalities are frequently seen in patients who have experienced a subarachnoid hemorrhage (SAH). These changes often mimic those seen in acute myocardial infarction although often patients have no history or other symptoms of cardiac disease. This paper reviews the types of rhythm disturbances seen following SAH and discusses the pathophysiologic basis for these changes. It also presents possible nursing interventions for these patients and explores areas needing further research. PMID- 2975314 TI - Neurosurgical treatment of intractable epilepsy. AB - Many patients with epilepsy are refractory to anticonvulsant therapy. For some, surgery is a means to control their seizures. Temporal lobectomy, the most common neurosurgical approach to intractable epilepsy, has resulted in improvement or cure in as many as 90% of treated individuals. To determine patients suitable for surgery, presurgical selection and evaluation criteria have been devised. With epilepsy surgery becoming more widely utilized and more centers appearing in the United States, there is a growing need for neuroscience nurses to know about recent developments. This overview of surgery for epilepsy includes the presurgical evaluation process through the postoperative phase presented from a nursing perspective. PMID- 2975315 TI - Movement therapy. AB - Nursing's holistic approach to patient care requires that nurses use interventions that promote holism. Movement therapy is one of a number of possible holistic interventions neuroscience nurses can use. This intervention helps persons to become aware of their bodies and to use the body for expressing inner feelings. Two techniques, creative dance and afspaending, are described along with suggestions for their use with neuroscience populations. PMID- 2975316 TI - A neurosciences internship and graduate nurse role conception. AB - Registered nurse (RN) turnover is a severe problem many hospitals face in today's health care climate. A study was conducted of a graduate nurse (GN) neurosciences internship established in a large metropolitan teaching hospital located in southwestern Tennessee to determine the internship's effect upon role conception of GNs. Thirty-two GNs enrolled in the internship comprised the experimental group, and 22 GNs in traditional orientation were the control group. Data collection instruments consisted of Corwin's Role Conception tool and a demographic questionnaire designed to examine variables that could influence professional role conception. Both instruments were completed at the beginning of employment, and Corwin's Role Conception repeated after completion of the first six months of practice. At this time audiotaped interviews were conducted by a nurse counselor to collect qualitative data from ten randomly selected informants from each group. Results revealed no significant differences in the two groups regarding changes in role conception as measured by Corwin's instrument. Audiotaped interviews brought to light many concerns of new GNs with regard to inadequate staffing, the charge nurse role, coping with the stress of the professional nurse role and new attitudes toward the field of the neurosciences. Implications for nurse administrators include the need to: be aware of the stresses to which new GNs are exposed; examine traditional practices such as the charge nurse role and staffing patterns; determine whether nursing internships, such as the neurosciences internship can improve the attitudes of GNs toward their specialty. PMID- 2975317 TI - Strategies for journal publication. AB - Publication in professional journals has become an expectation not only of neuroscience nurse educators but practitioners as well. Through publication, the neuroscience nurse demonstrates commitment to the profession and specialty area by sharing clinical insights and knowledge with colleagues practicing in the field. The publication process includes many steps beginning with the idea for publication to a careful review of the galley proofs. This article describes opportunities for journal publication and strategies for ensuring success in the publication process. PMID- 2975318 TI - The nurse's use of restraint with a neurologically impaired patient. PMID- 2975320 TI - QRS changes during acute ischemia induced by balloon occlusion of the LAD artery. PMID- 2975319 TI - False-positive posterior myocardial infarct criteria during left anterior descending coronary angioplasty. AB - In reviewing ECG change during acute anterior myocardial infarction (AMI) and during transluminal coronary angioplasty of the left anterior descending (LAD) coronary artery, QRS changes mimicking posterior myocardial infarct (PMI) and false-positive points for PMI using the QRS infarct size scoring system were observed in about 20% of patients. Nineteen patients in whom a three-lead electrocardiogram (ECG) was continuously recorded during transluminal coronary angioplasty of a proximal left anterior descending (LAD) coronary artery were reviewed. All but two had significant ST-segment shifts in V2 within a few seconds of balloon occlusion of a proximal LAD. In many such patients the R wave amplitude in V2 measured from the PR segment baseline remained relatively stable and the S amplitude tracked with the J. point. However, of the 19 patients studied, 11 had transient QRS changes greater than could be accounted for by baseline shift alone and 4 had transient false-positive PMI points by QRS MI size score. Transient QRS changes were observed as follows: (1) significant increase in R waves and/or decrease in S waves in V2 due to right septal block (or conduction delay) occurred in 3 of 11 patients and was the cause of the false positive criteria for PMI; (2) 5 of 11 patients had a major decrease in R waves in V2 due to left septal block; (3) 2 of 11 patients had both (1) and (2); and (4) 1 of 11 had transient frontal plane axis shifts with the QRS configuration of anterosuperior fascicular block plus right and left septal block.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975321 TI - Quantitative ST-segment recovery following angiographically successful angioplasty: a useful warning of early complications. AB - The angiographic/anatomic appearance of the TCA site and transstenotic gradient trending are two available intraprocedural variables that help to identify patients at high risk for early complications after successful angioplasty. We have reported on an additional variable, the rate of ST recovery following the final balloon deflation as a physiologic marker to identify patients at risk for early complications. Slow ST recovery was present in 52% of patients with early complications of myocardial infarction, urgent or emergent coronary bypass surgery, and/or death, whereas normal ST recovery was seen in 97% of patients with uncomplicated courses. ST trending is a non-invasive modality that is available in all patients undergoing TCA and should be a useful adjunct in identifying patients at high and low risk for early major complications following angiographically successful angioplasty. A prospective study of ST recovery during TCA deserves consideration. PMID- 2975322 TI - Poor performance of lead V5 in single- and dual-channel ST-segment monitoring during coronary occlusion. PMID- 2975323 TI - Relationship between Doppler left ventricular diastolic filling indexes and the electrocardiographic criteria for left atrial enlargement. AB - Traditional electrocardiographic (ECG) criteria for left atrial enlargement (LAE) emphasize the increased amplitude and width of the corresponding component of the hypertrophied atrium. Although a correlation exists between LAE and ECG criteria, a cause-and-effect relationship has not been conclusively demonstrated. Because the diastolic properties of the left ventricle directly influence left atrial emptying, these properties might also influence the ECG diagnosis of LAE. Therefore, the authors hypothesized that the ECG criteria for LAE are influenced by diastolic properties of the left ventricle as defined by Doppler-derived parameters. PMID- 2975324 TI - A systematic cytogenetic study of a population of 1170 mentally retarded and/or behaviourly disturbed patients including fragile X-screening. The Hondsberg experience. AB - A cytogenetic study was performed in a population of 1170 mentally retarded and/or behaviourly disturbed patients of the Hondsberg Institute in the south of the Netherlands. The cytogenetic data are presented and discussed. In all patients chromosomal evaluation was performed with Giemsa-banding and Quinacrine fluorescence, and additional banding techniques were performed whenever they were necessary to clarify the chromosomal abnormality. A fragile X screening with M199 cultures was performed in 311 males. In 22.1% of the patients a chromosomal basis was found for their developmental retardation: 14.3% Down syndrome patients, 6.1% other chromosomal abnormalities (mainly partial autosomal trisomies and monosomies and sex-chromosome abnormalities). In 24 males, through 21 index patients, a positive fragile X screening was found, i.e. 6.7% of the screened population and 1.8% of the total population. These results indicate that the diagnostic contribution of the fragile X screening is numerically of equal importance as are advanced chromosome banding techniques, and its contribution to the diagnosis of fragile X syndrome in one index male patient in general leads to the detection of several female relatives at risk to be carrier of this X-linked recessively inherited condition. The causal relationship between the occurrence of mental retardation and chromosomal aberration in genera i.e. autosomal trisomies, partial autosomal trisomies and monosomies, and Xq27-28 fragility is well established and is, to some extent, easy to understand. Whether carriers of other chromosomal rearrangements, mainly of balanced reciprocal and Robertsonian translocations, small extra chromosomes, paracentric inversions and chromosomal variants, have increased risk for mental handicap and/or congenital malformations in their progeny, remains unclear at the present time. Some of these residual problems and questions are discussed in the perspective of their importance for genetic counseling. Detailed data will be presented about the mental development and psychological profile of patients with these different types of chromosomal abnormalities and rearrangements. PMID- 2975325 TI - Portable dental practice: a new dimension for the practitioner. PMID- 2975326 TI - More on AIDS. PMID- 2975327 TI - Periodontal ultrasonic diagnosis: experiments on thin bony platelets and on a simulated periodontal ligament space. PMID- 2975328 TI - Histopathologic features of spontaneous periodontitis in adult Macaca fascicularis monkeys. PMID- 2975329 TI - Distribution of 3H-fucose in the transseptal ligament of the mouse. PMID- 2975330 TI - Evaluation of test statistics in split-mouth clinical trials. PMID- 2975331 TI - Flurbiprofen treatment of human periodontitis: effect on alveolar bone height and metabolism. PMID- 2975332 TI - Effect of lithium carbonate in HIV-infected patients with immune dysfunction. AB - Ten homosexual men received oral lithium carbonate at doses that maintained their serum lithium concentrations between 0.5 and 1.5 mEq/L. Prior to treatment all patients had HIV isolated from PHA-activated peripheral blood lymphocytes (PBLs) using a quantitative antigen-capture enzyme-linked immunosorbent assay (ELISA) assay for detection, and had an absolute number of CD4 (helper) lymphocytes of less than 300/mm3. Eight of 10 patients developed symptoms of drug toxicity requiring discontinuation of the drug in 7 patients. Two patients completed only 4-5 weeks of lithium therapy, and 5 patients received 7-8 weeks. All patients remained culture positive for HIV during the trial, and viral titers as measured by the antigen capture assay were unchanged or increased. There were no significant changes in the absolute number of CD4 lymphocytes, CD4/CD8 ratio, or phytohemagglutinin (PHA) or tetanus toxoid induced proliferative responses. There was a significant decrease in mixed lymphocyte reaction (MLR). Lithium carbonate demonstrated no immunorestorative or antiviral activity when given in therapeutic doses. Drug toxicity limited therapy in the majority of patients. PMID- 2975334 TI - Computer-aided instruction with the multiply impaired. PMID- 2975333 TI - [Studies on active constituents in medicinal plants using the receptor binding assay. II. Dopamine 2 receptor]. PMID- 2975335 TI - A comparison of two behaviour reduction procedures: traditional extinction alone and interpolated reinforcement followed by extinction. AB - Extinction responding of children with severe retardation was compared when one component of a multiple schedule (VR-20/VR-20) was interpolated with one session of additional reinforcement. Either variable ratio 10 (VR-10), variable ratio 5 (VR-5) or continuous reinforcement (CRF) were interpolated in one component prior to extinction in both. Interpolated reinforcement resulted in an extinction process at least as efficient as traditional methods. Total responding in the VR 10 and VR-5 components was less than in the comparable unchanged components. Results support and extend research with animals and humans. Results also suggest the feasibility of using such a procedure to gain control of behaviours that are controlled by intermittent schedules of reinforcement, are under multiple or inconsistent stimulus control or occur infrequently. PMID- 2975336 TI - Studies on experimentally induced hyperphenylalaninemia. AB - The administration of several compounds that can increase plasma phenylalanine levels and/or inhibit phenylalanine hydroxylase in rats was studied in order to determine their usefulness in inducing a phenylketonuria-like state. The results of this investigation revealed that 4.5 microns/10 g p-chlorophenylalanine is more effective than L-phenylalanine, alpha-methylphenylalanine, trimethoprim, Bactrim and Septra, since the former compound produced both adequate hyperphenylalaninemia and marked inhibition of hepatic phenylalanine hydroxylase activity. In addition, a 24-h study provided important insights into the changing diurnal patterns of specified biochemical parameters. PMID- 2975337 TI - Aortic regurgitation and mitral valve prolapse with Down's syndrome: a case control study. AB - This case-control study was designed to determine whether aortic regurgitation (AR) and mitral valve prolapse (MVP) are specifically associated with Down's syndrome, or occur in a comparably retarded population without Down's syndrome. The 92 control subjects resided at the same institution as the 131 cases, and had mental retardation attributable to low birth weight or perinatal asphyxia. Mean (+/- SD) ages of the cases and the control group were 41 +/- 11 years compared with 40 +/- 13 years, respectively. The relative risk of AR among the cases (those with Down's syndrome) was 2.9 (95% confidence interval, 0.65-13.2; P = 0.16) and that of MVP was 3.5 (95% confidence interval, 1.2-10.2; P = 0.02). The results showed that these two valvular abnormalities are specifically associated with Down's syndrome in adults. PMID- 2975338 TI - Bacteriophage lambda site-specific recombination proceeds with a defined order of strand exchanges. AB - Previous work has established that integration of the genome of bacteriophage lambda into the chromosome of its bacterial host proceeds via two independent strand exchanges, which make and then resolve a Holliday-structure intermediate. We find that a phosphorothioate substitution at the site of exchange in one strand of a recombination site depresses the yield of Holliday structures much more than a similar substitution in the other strand. Furthermore, we show that the Holliday structures that accumulate in unblocked reactions have all been made by recombination of one particular pair of strands. We conclude that there is a strong bias in the choice of strands that initiate crossing-over. Excision, the recombination reaction that excises the integrated prophage, exhibits the same bias as integration. This proves, at least at the level of strand exchange, that excision is not the simple reversal of integration. We have altered the relative orientation of parts of the phage attachment site, attP, to demonstrate that the strand-exchange bias is determined not by the local environment around the point of exchange in the core of attP but by more distant elements in its arms. This suggests that the order of the strand exchanges is dictated by an asymmetry in the way that the nucleosome-like structure that forms at attP brings the bacterial site, attB, into juxtaposition prior to strand exchange. Finally, we use the altered attP to show that homology between attP and attB is most critical when it is adjacent to the point of strand exchange. PMID- 2975339 TI - Chronic hypertension changes myosin isoenzyme pattern and decreases myosin phosphorylation in the rat heart. AB - We investigated systolic blood pressure (BP), ventricular myosin isoenzyme (MI) pattern, and myosin P-light chain phosphorylation (MP) of male and female normotensive (WKY) and spontaneously hypertensive rats (SHRSP). BP increased in SHRSP of both sexes during maturation. Male SHRSP reached a significantly higher BP (262 mmHg at week 64) than female SHRSP (217 mmHg at week 64). WKY remained at approximately 114 mmHg throughout the life-span investigated (5 to 64 weeks). MI pattern (expressed as %V1/%V3) shifted age-dependent to the V3 form: In female SHRSP MI pattern was 41/25 at week 18, 34/35 and 40/38 within week 22 to 32, and shifted to 18/53 until week 64. In male SHRSP MI pattern was 25/44 at week 18 and shifted gradually to 13/60 until week 53. MI patterns of WKY of both sexes were 100% V1 within week 5 to 12, shifted gradually to 51/23 and then remained constant until week 64. MP of the ventricle of female WKY and SHRSP was approximately 41% until week 52. At week 64, however, MP of female SHRSP decreased to 18% whereas female WKY remained at approximately 41%. MP of the ventricle of male WKY and SHRSP was approximately 38% until week 38. At week 44, however, MP of male SHRSP decreased to 22% whereas male WKY remained constant. Isometric tension generation of chemically skinned rat ventricular fibres increased after MP by calcium-calmodulin-dependent myosin light chain kinase. Both the shift to the V3 form and the decreased MP level might contribute to the development of cardiac failure in old SHRSP of both sexes. PMID- 2975340 TI - Oxygen diffusion distance in thyroxine-induced hypertrophic rabbit myocardium. AB - We studied the oxygen diffusion distance in the rabbit myocardium in untreated animals and in animals treated with thyroxine (T4) for 3 days and 16 days. The subepicardial and subendocardial regions were studied separately. Sixteen days of T4 treatment results in significant cardiac hypertrophy. After 16 days, the percentage of the myocardium occupied by myocytes decreases as the volume of extracellular matrix increases. After 3 days, the number of myocyte mitochondria per unit volume of myocardium increases without an increase in mitochondrial volume. After 16 days the volume density of the myocyte mitochondria increases. Oxygen diffusion distance was determined by measuring the distance between the myocardial capillaries and the cardiomyocyte mitochondria. The deposition of extracellular matrix results in an increase in the distance between the myocardial capillaries and the myocytes. The distribution of the mitochondria within the myocytes remains unchanged resulting in an increased distance between the capillaries and the mitochondria, hence, an increased oxygen diffusion distance. There is some evidence that the mitochondria most distant from the capillaries are the mitochondria in which division is most frequent with T4 stimulation. This may be related to the relative deprivation of oxygen of these mitochondria. PMID- 2975341 TI - Coronary vascular function and morphology in hydralazine treated DOCA salt rats. AB - The purpose of these studies was to evaluate structural and functional changes in a model of hypertension-induced cardiac hypertrophy in which vasodilator therapy prevented the increase in blood pressure. Uninephrectomized weanling (125 g) Sprague-Dawley rats received a Silastic implant containing deoxycorticosterone acetate (DOCA, 150 mg/kg) subcutaneously and were given drinking water containing sodium chloride and potassium chloride. Vasodilator antihypertensive treatment (hydralazine; HYD) was started immediately after DOCA implantation. The rise in blood pressure was prevented in DOCA + HYD (124 +/- 5.4 mm Hg, +/- S.E.M.) compared to DOCA (213 +/- 7.5 mm Hg), and blood pressure was not different from control (CON; 118 +/- 5.5 mm Hg). Hydralazine lowered blood pressure in CON + HYD (102 + 3.9 mm Hg) but this decrease was not significant (P greater than 0.05). Hydralazine treatment prevented hypertension in DOCA + HYD but did not prevent development of cardiac hypertrophy (heart weight/body weight of DOCA + HYD 3.99 +/- 0.1 vs. DOCA 4.15 +/- 0.1; CON, 3.23 +/- 0.2 and CON + HYD 3.27 +/- 0.1). Coronary flow reserve measured by adenosine vasodilatation in a modified Langendorff isolated perfused rat heart model, was decreased in hearts from DOCA rats (41% increase in flow above baseline) compared to controls (CON, 132%; CON + HYD 139%), and was significantly improved in DOCA + HYD (98%). Morphometric evaluation of perfusion-fixed coronary arteries demonstrated a significant increase in the slope of the regression line comparing the square root of medial area vs. outer diameter in DOCA (0.619) compared to CON (0.501) and CON + HYD (0.491). Blood vessels from DOCA + HYD were not different from control (0.503). These studies suggest that significant alterations in coronary vascular structure and function occur in hypertension-induced cardiac hypertrophy. The coronary vasculature is responsive to blood pressure, independent of cardiac hypertrophy, although coronary deficits do remain after antihypertensive therapy. PMID- 2975342 TI - Peripheral motoneuron interactions with laminin and Schwann cell-derived neurite promoting molecules: developmental regulation of laminin receptor function. AB - Schwann cells synthesize several neurite outgrowth-promoting molecules and localize them in either the extracellular matrix (ECM; e.g., laminin) or on the plasma membrane (e.g., L1/NgCAM and N-cadherin). Neurite outgrowth by embryonic chick ciliary ganglion (CG) neurons in response to these Schwann cell molecules largely depends on several specific neuronal cell surface receptors: integrin beta 1-class ECM receptors, L1/NgCAM, and N-cadherin (Bixby et al.: Journal of Cell Biology 107:353-361 1988). To address whether neuronal ECM receptors are regulated independently of cell surface adhesion molecules, we studied the ability of dissociated CG neurons from different developmental ages to extend neurites rapidly on 1) substrates coated with the ECM glycoprotein laminin (either from Schwann cell-conditioned medium or purified from the Engelbreth-Holm Swarm sarcoma) or 2) the surfaces of Schwann cells or Schwannoma (RN22) cells. CG neurons gradually lost the ability between embryonic day 8 (E8) and E14 to attach to and extend neurites in an integrin-dependent fashion on purified laminin or Schwann cell-derived laminin. The inability of E14 CG neurons to respond to laminin was partially reversed after explantation for 2.5 days in vitro, which increased the percentage of responsive neurons approximately ten-fold. E14 neurons remained capable of extending neurites rapidly on the surfaces of Schwann and Schwannoma cells. Thus, the inability of E14 neurons to respond to laminin reflects a specific loss of laminin receptor function, while other receptors, most likely N-cadherin and L1/NgCAM, remain capable of promoting neurite outgrowth on Schwann cell surfaces. Since integrin beta 1-class heterodimers have been shown to function directly as receptors mediating neuronal attachment and process outgrowth on laminin, our results imply that the expression or function of laminin-binding integrin heterodimers is regulated during the development of CG neurons. The apparent loss of integrin receptor function occurs during the period when the axons of CG neurons innervate their targets. Substantial integrin receptor function is recovered when target contact is disrupted by explantation. Thus, the functions of integrin-class receptors in CG neurons may be regulated by target contact. PMID- 2975343 TI - [Attribution of extracellular enzymes to bacterial infectious diseases]. PMID- 2975344 TI - [Diagnostic validity of hysterosalpingography and laparoscopy in infertile women]. PMID- 2975346 TI - [Hepatitis vaccination of health personnel]. PMID- 2975345 TI - Donated dental services gaining momentum in state. PMID- 2975347 TI - [Changes in T lymphocyte functions after UV irradiation]. PMID- 2975348 TI - [Therapy concepts in colorectal liver metastases. What is proven, what is open to discussion?]. AB - Isolated liver metastases of colorectal carcinomas principally can be treated by surgical intervention or cytostatic chemotherapy. Unquestionable indications for resection therapy are either solitary metastases or metastases limited to one liver lobe, since resection provides the best long-time results. In multilocular metastases or non-resectable liver metastases systemic monochemotherapy with 5 Fluorouracil (5-FU) presents respondance rates of about 20 percent with remissions lasting 3 to 6 months. Thereby a prolongation of life could not be proven statistically. Initial studies with a combination therapy of 5-FU/Folinic acid promise higher remission rates due to an increased cytotoxicity caused by a synergistic effect. Because of the mainly arterial supply of liver metastases the different procedures of regional chemotherapy-intraarterial infusion, isolated liver perfusion, chemoembolisation-provide the tumor with high drug concentrations without provoking systemic side effects. This advantage of a regional application of cytostatic drugs is reduced by the high percentage (2 to 87 percent) of extrahepatic tumor manifestations occurring after an average of 6 to 8 months. PMID- 2975349 TI - [Effect of cyclophosphamide on the growth of a transplantable sarcoma in the rat (S-E 100)]. PMID- 2975350 TI - [Bronchobiliary fistula in necrotizing liver metastasis of sigmoid cancer following regional chemotherapy]. PMID- 2975351 TI - Translational signals of a major head protein gene of bacteriophage lambda. AB - The D gene of bacteriophage lambda which codes for a major head protein is expressed at a high level during lytic growth. We have constructed a set of D lacZ gene fusions in order to examine the factors determining the high efficiency of the D translational initiation signals. It was found that an integral sequence, 300 bp long and upstream of the ATG initiation codon, is required for maximal protein synthesis. PMID- 2975352 TI - Onychomycosis due to a fungus of the Aspergillus versicolor group. PMID- 2975353 TI - Experimental candidiasis associated with liver injury. Role of transferrin. AB - In an attempt to perform a further investigation on the proposal that an increasing susceptibility to Candida infection in liver injury may be related to unsaturated transferrin level (UIBC) and/or to a total amount of transferrin represented by TIBC, we conducted experimental candidiasis using mice with galactosamine-induced liver injury and investigated the effect of preadministration of transferrin prior to inoculation of Candida albicans. Final mortality was 10% in the mice without liver injury and without transferrin (Group 1) and ones with liver injury and with transferrin (Group 3). By contrast a 50% mortality was given in one only with liver injury (Group 2). The TIBCs in Groups 1 and 3 were significantly higher than that in Group 2. The UIBCs in Groups 2 and 3, although there was no significant difference between them, were significantly lower than that in Group 1. This study confirmed that transferrin (TIBC) may have a direct deterring effect on systemic Candida infection and the decreased TIBC in the liver injury enhances the growth of C. albicans. PMID- 2975355 TI - [Incidence and results of re-interventions following coronary vessel surgery]. PMID- 2975354 TI - Molecular pharmacology of 5-HT1D recognition sites: radioligand binding studies in human, pig and calf brain membranes. AB - 1) The binding characteristics of [3H]5-HT (5-hydroxytryptamine, serotonin) were investigated in membrane preparations of several regions from calf, pig and human brain in the presence of 100 nmol/l 8-OH-DPAT (8-hydroxy-2[di-n dipropylamino]tetralin) and 100 nmol/l mesulergine in order to mask 5-HT1A and 5 HT1C sites. 2) [3H]5-HT bound rapidly, reversibly and stereo-selectively to a population of high affinity recognition sites in membranes from pig caudate, calf caudate and human cortex, caudate and substantia nigra. 3) Saturation experiments carried out with [3H]5-HT in the presence of 100 nmol/l 8-OH-DPAT and 100 nmol/l mesulergine revealed that non-5-HT1A non-5-HT1C sites represented from 50 to more than 90% of the total 5-HT1 sites (determined with [3H]5-HT in the absence of 8 OH-DPAT and mesulergine), depending on the tissue source. 4) The pharmacological profile of these sites was characterized in competition experiments performed with a variety of ligands in membranes of calf, pig and human caudate membranes. Under these conditions, [3H]5-HT labelled a population of "5-HT1-like" sites which display nanomolar affinity for tryptamines (5-carboxamidotryptamine greater than 5-HT greater than or equal to 5-methoxytryptamine greater than tryptamine) and some ergolines (metergoline greater than methysergide). In contrast, these sites showed low affinity for drugs with high affinity and/or selectivity for 5 HT1A (8-OH-DPAT, buspirone), 5-HT1B (21-009, RU 24969), 5-HT1C (mesulergine, mianserin) and 5-HT2 sites (ketanserin, cinanserin).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975356 TI - [Prevention of neurological symptoms due to atlanto-axial dislocation in persons with Down syndrome]. PMID- 2975357 TI - [Down syndrome, epiphysiolysis of the femoral head and hypothyroidism: a wellknown association?]. PMID- 2975358 TI - Immunocytometric characteristics of a monoclonal antibody (Bra55) recognizing the leukocyte common antigen (LCA). AB - The immunofluorescence cytofluorometric reactivity pattern of monoclonal antibody Bra55 (IgG1) elicited with a non-T, non-B ALL cell line (REH), with a panel of human neoplastic hemopoietic cell lines (including non-T, non-B, T and myeloid leukemia cell lines) and with isolated peripheral blood lymphocytes, monocytes and granulocytes from healthy donors corresponded to the previously described microscopic immunofluorescence, ELISA, immunoprecipitation and immunocytochemic data indicating that this monoclonal antibody recognizes a 170-220 kDa cell surface glycoprotein (leukocyte common antigen) expressed selectively on hemopoietic cells. The purified, FITC-conjugated Bra55 monoclonal antibody was effectively inhibited in its binding to the surface of LCA-positive cells by reference anti-LCA monoclonal antibodies; no inhibition of this activity by LCA unrelated monoclonal antibodies (such as anti-MHC class I and class II antibodies) was observed. These data confirm the previously reported hemopoietic cell specificity (anti-LCA, CD45) of the Bra55 monoclonal antibody. PMID- 2975359 TI - Identification of murine placental glycoproteins in relation to in vivo and in vitro immunomodulatory properties against H-2 antigens. AB - Glycoproteins were fractionated from placentae of syngeneic mice (CBA female xCBA male - H-2k) and evaluated in vivo and in vitro for the modulation of allo-immune responses against the H-2 incompatible A/J (H-2a) strain. It became apparent that the placenta contained high (309-800 kDA) and low (less than 13 kDA) molecular weight proteins, which favoured Sa1 allograft enhancement, leading to 75-100% lethal tumors, and inhibited mixed lymphocyte cultures (MLC). Though the low molecular weight fraction did not modify the mast cell degranulating (DAAD) antibody response, high molecular weight proteins caused a low production of DAAD allo-antibodies of the IgG1 + IgE isotypes. Moreover, addition of the placental fraction representing a specific band of 119 kDA resulted in the production of allo-immune sera rich in DAAD antibodies, which, however, were not associated with allograft enhancement. Beside these components, placenta is endowed with other proteins which modify humoral immune responses in different ways, as ascertained by the C-mediated cytotoxic antibody assay; fractions, rich in a 105 and 55 kDA bands, were immunosuppressive, whereas another, rich in a specific 100 kDA band, was active in eliciting enhanced production of alloantibodies of the IgG2 isotype. Moreover, the fractions representing specific bands of 50, 68 and 75 kDA, which were most effective in inhibiting MLC, neither caused lethal tumor enhancement nor modified IgG2 antibody responses. Based on in vivo and in vitro modulation of immune responses by placental products, it is concluded that: 1) allograft enhancement and high production of IgG1 antibodies are not linked to the same glycoprotein, 2) the immunomodulators in relation to the protection of viviparity appear to be located at the exclusion limits of Sephacryl S-200 (i.e. greater than 250 kDA and less than 13 kDA proteins) and 3) in vitro assays which depend on the inhibition of MLC by placental components should not be taken as the sole criterion for defining a given immunomodulator. PMID- 2975361 TI - Autoradiography of antidepressant binding sites in the human brain: localization using [3H]imipramine and [3H]paroxetine. AB - [3H]Imipramine and [3H]paroxetine were used to label sites associated with serotonin uptake mechanisms in post-mortem brain tissue from control subjects. The anatomical localization of these sites was examined by autoradiography and densities measured by microdensitometry. We found [3H]imipramine binding to increase with age in the cortex and amygdala, but to be independent of gender and post-mortem delay. Preliminary results indicate that the binding of both [3H]imipramine and [3H]paroxetine is diminished in the brain of patients treated with imipramine. The distribution of [3H]imipramine and [3H]paroxetine high affinity binding sites was very similar, and correlated well with the distribution of serotonergic presynaptic markers in the brain. The highest densities of binding sites were found in the raphe nuclei and the midline thalamic nuclei. Other structures presenting high levels of binding were the substantia nigra, nucleus interpeduncularis, locus coeruleus, nucleus nervi hypoglossi, nucleus nervi facialis, mammillary bodies and other parts of the hypothalamus. In contrast, regions such as the neocortex, hippocampus, amygdala and cerebellum showed low densities of [3H]imipramine and [3H]paroxetine binding sites. This distribution seems to indicate that the ascending serotonergic pathways are the main site of action of antidepressants. PMID- 2975360 TI - Regional distribution of dopamine, 5-hydroxytryptamine, and noradrenaline in the rat vas deferens. AB - The concentrations of dopamine (DA), 5-hydroxytryptamine (5-HT) and noradrenaline (NA) in the rat vas deferens divided in eight or four sections were determined by high performance liquid chromatography with electrochemical detection. Dopamine and NA had the same regional distribution; their concentrations were maximal near the prostatic end and decreased towards the epididymis. The concentration of 5-HT also decreased from the prostatic to the epididymal end, but 5-HT did not follow the same regional distribution as DA and NA. Reserpine (0.02 or 0.2 mg/kg, i.p., 24 hr) and 6-hydroxydopamine (2 x 80 mg/kg, i.v., 6 days) decreased the contents of DA and NA; the concentrations of both amines were modified to a similar extent. Reserpine also diminished the content of 5-HT. Pargyline (200 mg/kg, i.p., 2 hr) increased the concentration of 5-HT while p-chlorophenylalanine (300 mg/kg, oral, 3 days) decreased the contents of the amine in some sections of the vas deferens. This study suggests that DA and NA co-exist in the same sympathetic neurons. Some of the 5-HT could be stored in mast cells as previously proposed, but the finding that tissue content of 5-HT changes after inhibiting the deamination or synthesis of the amine suggests that other source(s) of 5-HT distinct from mast cells exist in the rat vas deferens. PMID- 2975362 TI - Crush accident resulting in isolated abdominal aortic injury. AB - The case history is presented of a patient with isolated abdominal aortic trauma due to blunt injury of the abdominal wall. Survey of the literature shows a typical clinical manifestation of this rare injury. When present, prompt recognition and proper treatment are essential in the management of this condition. PMID- 2975363 TI - [Disinfection of swimming pool water and its outcome]. PMID- 2975364 TI - [Use of ozone in the preparation of swimming and bathing water in West Germany]. PMID- 2975365 TI - [Determination of colony count in accordance with the drinking water regulation]. PMID- 2975366 TI - [Multilayer filtration in the preparation of bathing water--performance of the biofilter]. PMID- 2975367 TI - [Critical observations on suitability testing of synthetic pool liners]. PMID- 2975368 TI - [Contamination of cheese with Listeria monocytogenes]. PMID- 2975369 TI - [Gene technology--environmental health aspects of the release of microorganisms and viruses changed by genetic technics into the environment (the problem of release)]. PMID- 2975370 TI - [Report of experiences with a model for the assessment of long-term inpatient psychiatric cases in a large city by the appointed public health service--the Berlin model for long-term psychiatric patients]. PMID- 2975371 TI - [Supplementary data for cholesterol and blood glucose screening in public services of the Health Department]. PMID- 2975372 TI - Treatment outcome in low back pain patients: do compensation benefits make a difference? AB - Some evidence suggests that chronic pain patients who receive worker's compensation benefits have a tendency to exaggerate their symptoms and not benefit from treatment. This study compared 110 male chronic low back pain patients receiving either no compensation, time-limited compensation, or unlimited compensation on pretreatment and follow-up variables. The patients who received unlimited compensation tended to have a higher percentage of physician rated symptom dramatization, to have more pain behavior, and to use more medication than the no-compensation and time-limited compensation patients. At follow-up, fewer patients with unlimited compensation had returned to work as compared with the other groups. These results suggest that time-limited compensation may not affect treatment outcome or interfere with return to work, while unlimited compensation may adversely influence overall treatment outcome and the probability that patients will return to work. PMID- 2975374 TI - [Application of PTCA steerable guide wire for abdominal and peripheral interventional angiography]. PMID- 2975373 TI - [Granulocytic chalone and antichalone as homeostatic factors in the functional system of the blood in aseptic inflammation]. PMID- 2975375 TI - [Use of low-temperature plasma in the preparation of synthetic polymers for modification by collagen]. AB - The physico mechanical properties of films, vascular prostheses and nets made of polyethylene terephthalate and polyethylene film treated with the low temperature plasma were investigated. The plasma was generated by glow discharge in air under the following conditions: gas pressure - , current - 0.4-0.6 A, time of treatment - 40-240 s. stated that the angles of wetting with water and collagen solution of films treated with plasma decreases considerably. Sharp decreasing of wetting angles is followed by quick rise of the between water and polymeric film, and also by the increasing of free surface energy. The increasing of the wettability does not change during the storing time. The breaking stress and extension of the films treated with plasma are the same or a little bit higher than those of the untreated ones. The collagen modified by radiation shows high adhesion to the polymeric materials treated with plasma. The value of this adhesion 6 x 10(5) mN/m for polyester film, is about 6 times higher than the value of adhesion obtained by other authors, if the polyester film was treated with air plasma generated by spark discharge method. PMID- 2975376 TI - A curable cause of musculoskeletal pain. PMID- 2975377 TI - Injectable contraception. PMID- 2975378 TI - High dose ipratropium bromide--is it safe? PMID- 2975379 TI - [Workshop on "Asbestosis -assessment" in the Hospital for Accident Cases Hamburg Lohbrugge on 13 June 1987]. PMID- 2975381 TI - [Physical work capacity of agricultural workers with chronic bronchitis]. PMID- 2975380 TI - [Importance of db gene for the development of low dose streptozotocin diabetes in C57BL/KsJ mice]. AB - The paper is concerned with some data on the effect of the diabetic gene (db) on mouse sensitivity to streptozotocin (SC). Male mice aged 2-3 mos. of mutant C57BL/KsJ strain (genotypes: m+/+m, db+/+m, db+/+db) were used for investigation. Insulin-dependent diabetes mellitus was induced by intraperitoneal injections of streptozotocin at a daily dose of 40 mg/kg for 5 days. The structure and function of the insular apparatus were histologically assessed as well as by the blood level of insulin and glucose within 15 days after the start of the experiment. The earliest hyperglycemic reaction to SC was typical of mice, homozygous by the diabetic gene; they had normoglycemia at the time of treatment. In mice, heterozygous by the diabetic gene, a hyperglycemic reaction developed later after treatment. However by the end of the investigation it reached values which were typical of mice, homozygous by the diabetic gene, with basal normoglycemia. Mice, not carrying the diabetic gene, as well as homozygotes by this gene with basal hyperglycemia, possessed lesser sensitivity to SC. The expression of hyperglycemic reactions showed correlation with a degree of dystrophic changes and the development of lymphocellular infiltration in the pancreatic islets of mice with basal normoglycemia in low dose streptozotocin diabetes. The development of spontaneous hyperglycemia in homozygotes by the diabetic gene lowered their sensitivity to SC diabetogenic effects. PMID- 2975382 TI - [Status and prospects of the development of anti-tuberculosis services in the RSFSR]. PMID- 2975384 TI - [Present-day characteristics of tuberculosis]. PMID- 2975383 TI - [Epidemiological hazard of alcoholic patients with tuberculosis and the organizational measures to reduce it]. PMID- 2975385 TI - Synthesis and photosensitizing activity of a di-porphyrin ether. PMID- 2975386 TI - Protection against dihematoporphyrin ether photosensitivity. PMID- 2975387 TI - [Interventional radiology]. PMID- 2975388 TI - [Percutaneous transluminal angioplasty of carotid artery stenoses-- clinical, neuroradiologic and electrophysiologic findings before and following treatment]. PMID- 2975389 TI - Community care. PMID- 2975390 TI - [Indications of dental treatment measures for HIV-infected and AIDS patients]. PMID- 2975391 TI - [Newest results in AIDS research--of special interest to dentist. Are dental protective measures excessive?]. PMID- 2975392 TI - [Induction of prophage lambda during the irradiation of E. coli cells exposed to radiations of various LET]. AB - Induction of lambda prophage in lysogenic E. coli cells exposed to ionizing radiation of different LET was studied as a function of dose I(D). Activities of pleiotropic RecA protein were shown to contribute to the shape of the I(D) curve. The experimental data were fitted by the function I(D) = alpha D(1-exp(-D0 1.D]exp(-beta D). Inducibility alpha increased with increasing LET which was related to the increased incidence of DNA lesions being a SOS-system call. PMID- 2975393 TI - [A mathematical model of the dynamics of granulocytopoiesis in mammals]. AB - A mathematical model has been developed for the dynamics of granulocytopoiesis in mammals subjected to chronic irradiation. The model involves a chalones mechanism of haemopoiesis regulation and comprises 12 nonlinear differential equations. The simulation results agree with the experimental data concerning the dynamics of granulocytopoiesis in rats affected by radiation within a wide range of dose rates. PMID- 2975394 TI - [Mathematical modeling of the dynamics of postradiation injury and recovery of intestinal epithelium]. AB - A mathematical model for the dynamics of the crypt-villus system in irradiated mammals has been developed. The model involves a chalones mechanism of regulation crypt cell reproduction rate and represents a system of four nonlinear differential equations. The simulation results are in a good agreement with the experimental data obtained within a wide range of doses. PMID- 2975395 TI - [Seborrheic dermatitis and HIV infection]. PMID- 2975396 TI - [Participation of the B-endorphinergic system in changes in intestinal motility induced in situations of psychic stress. Experimental study in dogs]. PMID- 2975397 TI - [Laparoscopic hemostatic procedures in patients with coagulation disorders undergoing hepatic and splenic puncture biopsies]. PMID- 2975398 TI - [Recurrent diarrhea, epigastric pain radiating to the back]. PMID- 2975400 TI - Clouded crystal ball. PMID- 2975399 TI - Laser Doppler velocimetry of fingertips during heat provocation in normals and in patients with Raynaud's phenomenon. AB - Measurements of Laser Doppler flow were performed on the fingertip during heating in a group of normals and in a group of patients with different types of Raynaud's phenomenon. During heating a group of 24 normals was distinguished from a group of 29 primary Raynaud's patients. A group of 17 patients with secondary Raynaud's phenomenon due to scleroderma and 8 patients with arteriographically proven severe vascular occlusive disease of the fingers showed a significantly lower response in comparison to the two above-mentioned groups. Patients with or without slight or severe trophic skin lesions were identified. In patients with scleroderma the maximal vasodilation during heating was inversely related to the number of organs affected. In conclusion, measurements of the hyperaemia during heating may play a role in determining the degree of obstructive vascular involvement in Raynaud's phenomenon. PMID- 2975401 TI - Finding the anti-oncogene. PMID- 2975402 TI - Amniocentesis use and risk awareness: comparison of knowledge and beliefs among older gravida. PMID- 2975403 TI - Fissure sealant in permanent first molars after 10 years. AB - The aim of the present study was to evaluate fissure sealing of permanent teeth in children during a 10 year period. All completely erupted caries-free permanent first molars were sealed in 6 to 9 year-old children forming the patient population of one of the authors. 758 sealed fissures were controlled and followed up in the study. The fissure sealant material Delton was used and the treatments were performed according to the manufacture's recommendations. The sealings were controlled once a year and the follow-up period varied from one to 10 years. After 8 years, about 80 per cent of the sealed fissures showed total sealant retention and no caries. Another 16 per cent of the sealed occlusal surfaces showed partial retention and no caries. After 10 years only 5.7 per cent of the sealed occlusal surfaces showed caries or restorations. The results underline that fissure sealing is an effective treatment and has a low failure rate. PMID- 2975404 TI - Oral dental care of the cancer patient. PMID- 2975405 TI - The irradiated head and neck cancer patient in your office. PMID- 2975406 TI - Changes in factor VIII proteins after cardiopulmonary bypass in man suggest endothelial damage. AB - 16 patients undergoing coronary artery bypass grafting using cardiopulmonary bypass (CPB) had blood samples taken at various times before, during and up to 1 week after surgery for estimation of beta-thromboglobulin (BTG), alpha-1 antichymotrypsin (ACT), factor VIII procoagulant protein (VIII:C), von Willebrand factor antigen (vWF:Ag) and ristocetin co-factor (vWF:RiCoF). vWF:Ag and vWF:RiCoF rose during and following surgery in a different manner to ACT. At 1 week there was a significantly disproportionate rise in vWF:Ag compared to vWF:RiCoF which suggested a degree of pulmonary endothelial damage. Prostacyclin, which was administered to 8 of the patients during CPB, reduced platelet activation as measured by a reduction in the release of BTG and also attenuated the consumption of VIII:C. It had no effect on pulmonary endothelial damage as measured by the ratio of vWF:Ag to vWF:RiCoF. PMID- 2975407 TI - Evidence for an increased generation of prostacyclin in the microvasculature and an impairment of the platelet alpha-granule release in chronic renal failure. AB - The formation of prostacyclin (PGI2) and thromboxane A2 and the release of beta thromboglobulin (beta-TG) at the site of platelet-vessel wall interaction, i.e. in blood emerging from a standardized injury of the microvasculature made to determine bleeding time, was studied in patients with end-stage chronic renal failure undergoing regular haemodialysis and in normal subjects. In the uraemic patients, levels of 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) were 1.3 fold to 6.3-fold higher than the corresponding values in the control subjects indicating an increased PGI2 formation in chronic uraemia. Formation of thromboxane B2 (TxB2) at the site of plug formation in vivo and during whole blood clotting in vitro was similar in the uraemic subjects and in the normals excluding a major defect in platelet prostaglandin metabolism in chronic renal failure. Significantly smaller amounts of beta-TG were found in blood obtained from the site of vascular injury as well as after in vitro blood clotting in patients with chronic renal failure indicating an impairment of the alpha-granule release in chronic uraemia. We therefore conclude that the haemorrhagic diathesis commonly seen in patients with chronic renal failure is--at least partially--due to an acquired defect of the platelet alpha-granule release and an increased generation of PGI2 in the microvasculature. PMID- 2975408 TI - The anticoagulant properties of a modified form of protein S. AB - Protein S (PS) is a vitamin K-dependent anticoagulant that acts as a cofactor to activated protein C (APC). To date PS has not been shown to possess anticoagulant activity in the absence of APC. In this study, we have developed monoclonal antibody to protein S and used to purify the protein to homogeneity from plasma. Affinity purified protein S (PSM), although identical to the conventionally purified protein as judged by SDS-PAGE, had significant anticoagulant activity in the absence of APC when measured in a factor Xa recalcification time. Using SDS PAGE we have demonstrated that prothrombin cleavage by factor Xa was inhibited in the presence of PSM. Kinetic analysis of the reaction revealed that PSM competitively inhibited factor Xa mediated cleavage of prothrombin. PS preincubated with the monoclonal antibody, acquired similar anticoagulant properties. These results suggest that the interaction of the monoclonal antibody with PS results in an alteration in the protein exposing sites that mediate the observed anticoagulant effect. Support that the protein was altered was derived from the observation that PSM was eight fold more sensitive to cleavage by thrombin and human neutrophil elastase than conventionally purified protein S. These observations suggest that PS can be modified in vitro to a protein with APC independent anticoagulant activity and raise the possibility that a similar alteration could occur in vivo through the binding protein S to a cellular or plasma protein. PMID- 2975409 TI - Activated protein C increases fibrin clot lysis by neutralization of plasminogen activator inhibitor--no evidence for a cofactor role of protein S. AB - The effect of purified human activated protein C (APC) on fibrinolysis was studied using a clot lysis system consisting of purified glu-plasminogen, tissue type plasminogen activator, plasminogen activator inhibitor (released from endothelial cells or blood platelets), fibrinogen, 125I-fibrinogen and thrombin. All proteins were of human origin. In this system APC could increase fibrinolysis in a dose dependent way, without affecting fibrin formation or fibrin crosslinking. However, this profibrinolytic effect of APC could only be observed when plasminogen activator inhibitor (PAI-1) was present. The effect of APC was completely quenched by pretreatment of APC with anti-protein C IgG or di isopropyl-fluorophosphate. Addition of the cofactors of APC-protein S, Ca2+-ions and phospholipid-alone or in combination did not enhance the profibrinolytic effect of APC. These observations indicate that human APC can accelerate in vitro clot lysis by the inactivation of PAI-1 activity. However, the neutralization of PAI-1 by APC is independent of the presence or absence of protein S, phospholipid and Ca2+-ions. PMID- 2975410 TI - Is AIDS a communicable disease? PMID- 2975411 TI - [Analysis of the pattern of nephropathies based on data from a nephrological center]. PMID- 2975412 TI - [Experience in serving the child population in hard-to-reach areas of Kazakhstan]. PMID- 2975413 TI - [Experience with the ophthalmological examination of workers in refrigerator manufacturing plants]. PMID- 2975414 TI - Parasites of the stomach in donkeys of the highveld of Zimbabwe. AB - Fourteen donkeys from a communal area of the Zimbabwean highveld were examined for stomach parasites during July and November of 1986 and January and April of 1987. All 14 animals were infected. Habronema muscae occurred in 12, Draschia megastoma in 11, H. majus in 9, Trichostrongylus axei in 9, Gasterophilus intestinalis in 14 and G. nasalis in 1 donkey. The lowest level of infection was in January for the helminths and in April for Gasterophilus larvae. The peak burdens of parasites were in July for Habronema spp. and in November for D. megastoma and Gasterophilus larvae. Infections with spirurid worms and Gasterophilus develop mainly from the middle of the rainy season to the beginning of the dry season (January to May/June). G. intestinalis seems to have an annual cycle. PMID- 2975415 TI - [The importance of repeated health resort treatment in increasing the efficacy of rehabilitation of patients with chronic bronchitis]. PMID- 2975416 TI - [Evaluation of the efficacy of sanatorium treatment of children with recurrent bronchitis using the factor analysis method]. PMID- 2975417 TI - [Diluted baths with volcanic mud in the therapy of patients with rheumatoid arthritis]. PMID- 2975418 TI - [Concentrated sunlight in the therapy of patients with rheumatoid arthritis]. PMID- 2975419 TI - [Natural radon waters of the Moscow area]. PMID- 2975420 TI - [Types of cellular antitumor cytotoxicity]. PMID- 2975421 TI - [The results and prospects of the automated follow-up of the dispensary care of cancer patients]. PMID- 2975422 TI - [Natural killer cells and T-suppressors in patients with chronic gastritis]. AB - In of 69 patients suffering chronic gastritis, 23 contracting gastritis in old age and 10 with gastritis aggravated by stomach ulcers, a significantly low level of natural killers was found in cases of mucosal atrophy and gastric intestinal metaplastic lesions, particularly, at an age of more than 60 years. T-suppressor level was relatively lower in patients with atrophic gastritis. PMID- 2975423 TI - [The role of an immunologic mechanism in the occurrence of postoperative metastases in mice]. AB - A comparative analysis of metastatic spread into lungs after tumor removal in immunologically normal mice (group 1) and those with splenectomy, thymectomy and congenital athymia-induced immunity disorders (group 2) was conducted. The largest number of lung metastases was found after tumor removal in group 1, the smallest in nude mice (group 2). The use of any method of lowering T-lymphocyte level (splenectomy, thymectomy or a single administration of low-dose cyclophosphamide) was shown to result in a significantly smaller number of lung metastases after tumor removal. It is suggested that formation of metastases after tumor removal is caused by an immunological factor which is possibly associated with an active response of T-suppressors. PMID- 2975424 TI - [The status of cancer care in the Chuvash ASSR]. AB - Recent years have witnessed an improvement in cancer patient registration procedures (due to the introduction of the automated system for cancer information processing), cancer control establishment network and the level of oncological awareness of general practitioners. The need of taking fresh measures aimed at improving diagnosis of malignancies of the stomach, breast and cervix uteri which are the leading factors of morbidity and mortality in the Chuvash ASSR is discussed. PMID- 2975425 TI - [Damage to the major vessels during surgical interventions in patients with cancer]. AB - The paper discusses causes and outcome of injuries to major arteries and veins observed in 53 surgical cases of cancer of various sites. Radical removal of tumor invading the vascular wall proved the main cause of iatrogenic trauma. Lateral and circular atraumatic sutures were used to restore major circulation in 31 cases; vessels were ligated in 17. Failure to arrest intraoperative bleeding was reported in 5 patients. There were 7 (13.2%) fatalities. PMID- 2975426 TI - [Ways to rationalize the nutrition of workers in the paint and varnish industry]. PMID- 2975427 TI - [The vitamin allowance and vitamin requirements of the operators of a chemical plant in the city of Rustavi]. AB - Excretion of thiamine, riboflavin, N1-methyl nicotinamide and ascorbic acid with urine, TDP-effect, ascorbic acid and vitamin A content in the blood have shown higher vitamin requirements in the operators, as compared to subjects of the control group, this was caused by unfavourable effects of the chemical factors of the production environment. The levels of the biochemical parameters characterizing normal providing of the operators with vitamins could be achieved by additional vitamins administered to the workers (2 dragees of "Hexavitum"). PMID- 2975428 TI - [Mineral composition of watermelon pulp in relation to the degree of ripeness and the region of its cultivation]. PMID- 2975429 TI - [Experience with the organization of child nutrition in an industrial city]. PMID- 2975430 TI - [Viral infection markers in foci of chronic hepatitis B]. AB - The intensity of detection and time course of the main markers of virus infection were studied in subjects who had contacts with patients in 74 foci with chronic active hepatitis, 52 with chronic persisting hepatitis and 41 with chronic hepatitis of unknown etiology. A high (up to 80.7%) frequency of detection of HBsAg, anti-HBs, anti-HBc was established in contact subjects which showed a trend of growth with the increase of the time of their contact with the source of infection. PMID- 2975431 TI - [Use of a continuous human T-lymphocyte H9 culture for screening chemical preparations that suppress the reproduction of the human immunodeficiency virus]. AB - Studies were carried out on the effectiveness of ribamidil, sodium phosphonoformate, cortisone, and deksazon as inhibitors of human immunodeficiency virus (HIV) reproduction which was tested by immunofluorescence. The drugs were screened in H9/IIIB culture permissively infected with HIV virus, and in H9 culture subsequently infected with cell-free virus-containing culture fluid or by adding live cells from H9/IIIB culture. The latter variant was shown to be advantageous, especially if the available cultures did not produce large amounts of the virus. Because of the effectiveness of the immunosuppressants, the possibility of screening in lymphocyte cultures and selection of preparations exerting no true antiviral effect is discussed. PMID- 2975432 TI - [The frequency of detection of HBs antigen, anti-HBs and anti-HBc in the healthy population of the Moldavian SSR]. PMID- 2975433 TI - [The dynamics of humoral immunity to the tick-borne encephalitis virus in the population of 3 regions]. PMID- 2975434 TI - Studies on lymphocyte subpopulations and cell-mediated immunity in patients with chronic renal failure. PMID- 2975436 TI - [75th anniversary of the Department of Skin and Venereal Diseases of the Saratov Medical Institute]. PMID- 2975435 TI - [Course over several decades of echinococcosis with peritoneal and pulmonary spread]. AB - Report on a 67-year-old man with echinococcus-granulosus-cysts in liver, lungs, spleen and mesentery since at least 31 years. In 1969 and 1974 surgical intervention was performed. In 1983 suppurative colliquation and spontaneous depletion by a fistula of a cyst of the liver, output of the cysts of the right lung by coughing. In 1986 spontaneous resorption of a cyst of the left lobe of the liver. In 1987 measurable shrinking of a cyst of the left lower lobe of the lung. Two until today existing cysts (spleen and upper part of the abdomen in the midline) are in supervision by sonography. PMID- 2975437 TI - [Characteristics of 3H-paroxetine-labeled serotonin uptake sites in rat brain]. AB - Paroxetine (PAR) is a potent and selective inhibitor of serotonin uptake, and it has been demonstrated that 3H-PAR is a favorable candidate for labeling of the serotonin transport system. In this report, the binding of 3H-PAR to rat brain membranes was further investigated to obtain a better understanding of its role in the neuronal serotonin-uptake mechanism. Consistent with previous reports, it is indicated that 3H-PAR binds with high affinity to a site closely related to the serotonin-uptake mechanism. This binding is highly sodium dependent. The finding that depletion of brain serotonin content with p-chlorophenylalanine did not induce any alteration in the number and the affinity of the 3H-PAR-binding sites suggests the lack of serotonergic modulation on these sites. Repeated administration of desipramine, clomipramine, mianserin or deprenyl also failed to change 3H-PAR-binding sites in cerebral cortex and hippocampus. Such findings are compared to previous studies on the serotonin-uptake mechanism and the site labeled by 3H-imipramine, and discussed in respect to the biochemistry of affective disorders. PMID- 2975438 TI - [The modifiability of periodontal tissue in Wistar rats by a variation in the magnesium content of the diet]. PMID- 2975439 TI - [Dental caries and the pH value of the saliva in children and juveniles with insulin-dependent diabetes]. PMID- 2975440 TI - [The mesiodens anomaly with a reversion to older dentin forms]. PMID- 2975441 TI - [Micromorphologic research on caries-dependent dentinal hypermineralization]. PMID- 2975442 TI - [Differential diagnostic delimitation of healthy gingiva and gingivitis (the initial lesion)]. PMID- 2975443 TI - [Electrothermographic measurements of the alveolar process during tooth eruption]. PMID- 2975444 TI - [The problems of treating mandibular fractures in the case of partial dentition or an edentulous jaw]. PMID- 2975445 TI - [Diagnosis of radiogenic jaw bone damage. The first experimental results with an animal model]. PMID- 2975446 TI - [Ultrahistochemical differentiation of the plaque matrix on 2 dietary regimens]. PMID- 2975447 TI - [Experimental materials research on the combination strength behavior of various opaque materials for a dental metal-plastic composite]. PMID- 2975448 TI - Reactogenicity and immunogenicity of a new recombinant yeast-derived hepatitis B vaccine. AB - Under randomized double-blind conditions, 220 medical students were vaccinated with either a 2.5, 5, 10, or 20 micrograms dose of a recombinant yeast-derived hepatitis B or a 20 micrograms dose of a plasma-derived vaccine. Vaccines were administered at months 0, 1, and 2. After 11 months, all vaccinees received a 20 micrograms booster dose of the recombinant vaccine. There were no significant differences in adverse reactions between the study groups. Induction of IgE antibodies to yeast was not observed. One month after the third vaccination, seroconversion rates reached 100% in all vaccines. Mean anti-HBs levels varied between 150 and 1470 IU/l after 3 vaccinations, with the lowest dose resulting in the lowest titres. Following the booster vaccination, dose-dependent effects were no longer observed. Anti-HBs concentrations were reanalyzed 29 and 36 months after the study had started. The data indicate that the recombinant hepatitis B vaccine is safe and immunogenic for use in man and comparable to the plasma derived vaccine in terms of safety and efficacy. PMID- 2975450 TI - [Work experience in optimizing epidemiologic health surveillance of intestinal infections at the provincial level]. PMID- 2975449 TI - [An outbreak of Legionella infection in Armavir]. AB - In March-April 1987 an outbreak of Legionella infection was registered among 236 workers at a rubber factory in Armavir. An ARD-like syndrome and acute alveolitis were the main variants of the clinical course of the infection. Legionella infection was diagnosed on the basis of a fourfold and greater increase in the titers of antibodies to L. pneumophila, serogroup 1, as determined in the indirect immunofluorescence test and enzyme immunoassay (EIA). In urine samples obtained from patients Legionella antigen was determined with the use of EIA techniques. The culture of L. pneumophila, serogroup 1, was isolated from the recycling-type water supply system of the factory. The outbreak was stopped by a complex of sanitary, hygienic and epidemic control measures. PMID- 2975451 TI - [Isolation of strains of a new ecological variant of Staphylococcus aureus from asses]. AB - 47 coagulase-positive staphylococcal strains were isolated from the nasal smears of 175 healthy donkeys. In accordance with the schemes of Akatov--Devriese and Mayer-Witte--Akatov, 10.6% of the cultures were classified with the coagulase positive species S. hyicus and 89.3%, with the species S. aureus. Out of S. aureus strains, 11.9% were found to have the characteristics of ecovar hominis, while 16.2% of the cultures could not be classified with definite ecovars. Most of the strains (71.4%) were found to differ from the known ecovars of S. aureus in their biological properties. For this reason, the above strains were classified with the new ecovar asinae. The authors propose to make the existing S. aureus identification scheme (the scheme of Mayer-Witte--Akatov) more complete by adding the tests for hyaluronidase and phosphatase. PMID- 2975452 TI - [Characteristics of the clinical and immunologic safety of inactivated influenza vaccines in children undergoing multiple immunizations]. AB - In a strictly controlled epidemiological trial on 12,643 school children aged 11 14 years the reactogenic properties and safety of killed influenza chromatographic vaccine under the conditions of multiple immunization were studied. A single immunization dose of the vaccine (0.2 ml) contained the hemagglutinins of influenza viruses A/Philippines/82 (H3N3) and A/Kiev/59/79 (H1N1), 3.5 micrograms each. The preparation was introduced by means of a jet injector. The vaccine was shown to be clinically and immunologically safe under the conditions of the regular multiple immunization of children over the period of 4 years. PMID- 2975453 TI - T-cell acute lymphoblastic leukemia with severe leukopenia: evidence for suppression of myeloid progenitor cells by leukemic blasts. AB - A patient with T-cell acute lymphoblastic leukemia presented with leukopenia due to neutropenia, no circulating blasts and normal hemoglobin level. Marrow leukemic T lymphoblasts inhibited in vitro normal CFU-GM colony growth and released an activity that stimulated normal BFU-E growth. This patient demonstrates that immature T cells may modify hematopoietic stem cell growth both in vitro and in vivo. PMID- 2975454 TI - The maternal-infant center in the control of hepatitis B. AB - A seroepidemiologic survey of HBV infection was carried out in four population groups served by the Maternity Hospital of Majorca, an island in the Mediterranean. The prevalence of HBV markers (HBsAg, anti-HBs, anti-HBc), determined by enzyme-immunoassay, reached 17.2% among 864 parturients, 17.2% in 783 hospital personnel, 2.7% and 19.2% among the 186 children and 73 husbands of parturients with no HBV markers, 22.8% and 43.8%, respectively, among the 44 children and 48 husbands of HBsAg chronic carrier parturients. The prevalences of HBsAg in the same groups reached 0.9%, 1.5%, 0% and 0%, 11.4% and 2.1%, respectively. The carrier state represents 50% of all infection cases among children and 4.8% among husbands of carrier women. The prevalence of markers, which was especially high among personnel working in maintenance services (39.1%), on surgical wards (35.3%), among midwives (27.6%) and in cleaning and laundry services (27.1%), and laboratories (21.4%), is used as an indicator in defining priorities for the vaccination of people at risk. Sixty-seven newborn babies of HBsAg carrier mothers were administered one dose of HBIG and three doses of hepatitis B vaccine; at 12 months of age, the seroconversion rate was 98.5%, whereas 1 child (1.5%) became a chronic carrier. Maternity hospitals represent the ideal centers in which to carry out the detection of HBsAg in pregnant women prior to parturition, to ensure that all newborns of chronic carrier women receive passive-active immunization, and to screen their family contacts and vaccinate those who are found susceptible to the infection. PMID- 2975455 TI - Fetal duodenal obstruction. A high risk indicator for Down's syndrome. AB - Seven cases of fetal duodenal obstruction were diagnosed during an ultrasound examination. In 5 out of the 7, Down's syndrome was later diagnosed. The finding of duodenal obstruction in a fetus is a high risk indicator for Down's syndrome. We therefore recommend prenatal cytogenetic examination to be performed in such cases. PMID- 2975456 TI - [Effect of panaxadiol saponin and panaxatriol saponin on the activity of ATPase in rabbit striatum microsome]. PMID- 2975458 TI - Atrial natriuretic peptides and renin release. AB - The relationship between endogenous plasma concentrations of atrial natriuretic peptide and renin was examined in resting normal subjects and patients with cardiac impairment. To test the hypothesis that atrial natriuretic peptide inhibits renin secretion, intravenous infusions of atrial natriuretic peptide were administered to normal volunteers, patients with end-stage renal failure, and conscious dogs in both sodium-replete and sodium-depleted states. Plasma atrial natriuretic peptide and renin were inversely related in normal subjects (r = -0.52, n = 140, p less than 0.001), but a weak positive association between these two variables was observed in patients with cardiac impairment (r = 0.32, n = 60, p less than 0.02). Low doses of both 26- and 28-amino-acid human atrial natriuretic peptide (2 pmol/kg/minute for two hours) given to sodium-replete normal subjects halved plasma renin compared with time-matched placebo values (19 +/- 4 and 18 +/- 3 versus 36 +/- 8 microU/ml, p less than 0.001 for both). Incremental doses of synthetic atrial natriuretic peptide suppressed plasma renin below time-matched placebo values in both sodium-replete (maximal suppression 1.2 +/- 0.4 versus 8.6 +/- 1.4 microU/ml, p less than 0.001) and sodium-depleted (maximal suppression 18.9 +/- 4.9 versus 51 +/- 13 microU/ml, p less than 0.05) dogs. This effect was initially apparent at low doses of atrial natriuretic peptide (1 pmol/kg/minute), and renin suppression was maximal, in both states, with lesser doses of atrial natriuretic peptide than those at which maximal natriuresis was observed. Atrial natriuretic peptide administered to patients with end-stage renal failure (10 pmol/kg/minute for one hour) caused no change in plasma renin. These data confirm that atrial natriuretic peptide inhibits renin secretion in a dose-related manner and suggest that this action of the peptide is modified by both the baseline sodium status and renal function of the recipient. PMID- 2975457 TI - Inhibitory effect of methyl 7-butyl-4,5,6,7-tetrahydro-3-methylamino-4,6-dioxo-5 propyl-2H-pyrazol o[3, 4-d]pyrimidine-2-carboxylate (AA-2379) on type III allergic (Arthus) reaction. AB - Methyl 7-butyl-4,5,6,7-tetrahydro-3-methylamino-4,6-dioxo-5-propyl-2H- pyrazolo[3,4-d]pyrimidine-2-carboxylate (AA-2379), a non-steroidal, non-acidic agent, markedly inhibits type III allergic (Arthus) reaction; the ID50 values of AA-2379 in the rat reversed passive Arthus pleurisy, the rat active Arthus pleurisy, and the reversed passive Arthus reaction in rat skin were 5-10 mg/kg, p.o., and 30 mg/kg of AA-2379 inhibited the active Arthus reaction in rabbit skin by about 50%. Dexamethasone, but not acidic non-steroidal anti-inflammatory drugs and aminopyrine, inhibited the Arthus reaction. The vascular permeability in the reversed passive Arthus pleurisy is enhanced biphasically in the early response mediated by physiologically active amines, prostaglandins, and leukotrienes, and in the late response mediated by complements and polymorphonuclear leukocytes (PMNs). AA-2379 inhibited the late response more potently than the early one. Furthermore, when given after the early response was reduced, AA-2379 obviously inhibited the late response. Rat zymosan-induced paw edema and mouse zymosan activated serum-induced peritonitis, mediated by complements, were dose dependently inhibited by AA-2379; the ID50 values were 11.4 and 10.2 mg/kg, p.o., respectively. The results suggest that AA-2379 differs from non-steroidal anti inflammatory agents in strongly inhibiting the late response of the Arthus reaction, which associated with PMNs. PMID- 2975459 TI - Myocardial effect of converting enzyme inhibition in hypertensive and normotensive rats. AB - The effects of converting enzyme inhibition on the cardiac mass, isomyosins polymorphism, and collagen network in the left ventricle have been studied in renovascular, hypertensive, spontaneously hypertensive, and normotensive rats. The isoenzyme profile of left ventricular myosins was used as an indirect marker of the intrinsic property of contractility, whereas the collagen network, measured by a morphometric method, represented an indirect structural marker of the arrhythmogenic risk. One-clip, two-kidney renovascular hypertension was associated with cardiac hypertrophy, a shift in the isomyosin profile, and accumulation of collagen within the left ventricular myocardium. In this renin angiotensin-dependent model, one month of treatment with converting enzyme inhibitor normalized blood pressure and consistently reversed cardiac hypertrophy and the isomyosin profile. Converting enzyme inhibitor treatment of 12-week-old spontaneously hypertensive rats for three months significantly decreased blood pressure but did not completely normalize it. The increase in cardiac mass observed in spontaneously hypertensive rats was not reversed by this short treatment. Nevertheless, the percentage of the V1 form of myosin increased slightly after treatment, and the collagen content of the left ventricle was considerably decreased. Converting enzyme inhibition did not decrease blood pressure in DOCA-salt hypertension, and no changes were observed in cardiac hypertrophy, isomyosin profile, or the collagen network. The cardiac hypertrophy that occurs with aging in normotensive rats was associated with a significant shift in isomyosin profile and a large accumulation of collagen. Thus, aging mimics several of the quantitative and qualitative changes in the left ventricular protein profile observed in hypertension. In young normotensive rats, converting enzyme inhibition significantly decreased blood pressure and left ventricular mass, increased the percentage of V1 isomyosin, and prevented the accumulation of collagen. In one-year-old normotensive rats, treatment for six months with converting enzyme inhibitor decreased blood pressure, decreased cardiac mass, and prevented the accumulation of collagen; the isomyosin profile was not modified. Converting enzyme inhibition, by acting on cardiac afterload, can bring about quantitative and qualitative changes in the cardiac proteins of both hypertensive and normotensive rats. PMID- 2975461 TI - Relation between cardiac hypertrophy and forearm vascular structural changes before and during long-term antihypertensive treatment. AB - In patients with hypertension, structural changes develop in the heart and in the systemic arteries that have a significant role in the maintenance and gradual worsening of the hypertensive disease. Blood pressure, basal and post-ischemic "maximal" forearm blood flow (strain-gauge venous occlusive plethysmography), and echocardiographic left ventricular mass index were measured in 28 hypertensive patients (WHO class I or II, 23 men and five women, aged 26 to 59 years). Minimal vascular resistance (mean arterial pressure/peak blood flow) was taken as an index of vascular structural changes. The same measurements were made in a group of 14 patients before and after long-term antihypertensive treatment: in eight patients after six and 12 months of treatment with captopril (50 mg twice a day, plus 25 mg of hydrochlorothiazide per day if necessary) and in six patients after six months of treatment with nitrendipine (20 mg per day, plus 50 mg of atenolol per day if necessary). A significant but weak direct correlation was found between the degree of left ventricular hypertrophy and mean arterial pressure (r = 0.41) or minimal vascular resistance (r = 0.31). Thus, patients were categorized according to whether they had left ventricular hypertrophy or impaired blood flow; the results suggested that left ventricular hypertrophy may be detected earlier than increased minimal vascular resistance. After six months of treatment, both captopril and nitrendipine significantly reduced left ventricular mass index and minimal vascular resistance. Left ventricular mass index was normalized in 50 percent of the patients, whereas minimal vascular resistance was normalized in one patient only. After 12 months of treatment, left ventricular mass index was normalized in all patients; minimal vascular resistance was on the average further reduced but normalized in only one additional patient. Thus, regression of cardiovascular structure also seems to occur earlier in the heart. PMID- 2975460 TI - Left ventricular hypertrophy in hypertension. Changes in isomyosins and creatine kinase isoenzymes. AB - In hypertension, the heart of small mammals can express different isoenzymic forms of proteins under the influence of overload and other modulating factors. The increase in ventricular mass is generally paralleled by progressive changes in the isoforms of at least two proteins that are involved in the contraction process, namely, myosin and creatine-kinase. This review summarizes the biochemical and molecular changes occurring during progression and with regression of cardiac hypertrophy in rats, humans, and other animals, and focuses on the role played by antihypertensive drugs in modulation of ventricular isomyosins. The implications of these observations for humans remain to be fully determined. PMID- 2975462 TI - Beneficial effects of one year's treatment with captopril on left ventricular anatomy and function in hypertensive patients with left ventricular hypertrophy. AB - This study evaluated by echocardiography (M-mode tracings, two-dimensional guided) the effects of captopril administration for one year (25 to 50 mg twice a day, alone or in combination with hydrochlorothiazide when necessary) on left ventricular mass index, on systolic function at rest and during stress (hand grip for three minutes and cold pressor test for three minutes), and on diastolic function in 15 patients with essential hypertension (13 men and two women, aged 30 to 67 years) with left ventricular hypertrophy. In addition, supine and standing plasma catecholamine concentrations, plasma renin activity, and plasma aldosterone levels were measured. Examinations were performed during a placebo period and after three, six, and 12 months of captopril treatment. Blood pressure was significantly reduced (p less than 0.001), but heart rate did not change. Left ventricular hypertrophy was progressively reduced during treatment, mainly through reduction of left ventricular wall thickness. After one year, all patients had a normal left ventricular mass index (less than 120 g/m2). Before and during treatment, left ventricular systolic function, at rest and on maximal hand grip and cold pressor testing, evaluated on the basis of fractional shortening as related to end-systolic stress, was within the 95 percent confidence limits (calculated in a group of 25 normal subjects) in all 15 patients with essential hypertension. The percent increase in left ventricular dimensions during the diastolic rapid filling phase was significantly increased by treatment (p less than 0.05), indicating improvement of left ventricular relaxation. As expected, plasma renin activity was increased, plasma aldosterone levels were decreased, and plasma catecholamine concentrations did not change. These results indicate that long-term treatment with captopril has beneficial effects on left ventricular anatomy and function in patients with essential hypertension. PMID- 2975463 TI - Does the renin-angiotensin-aldosterone system modify cardiac structure and function in essential hypertension? AB - To determine the impact of the renin-angiotensin-aldosterone system on left ventricular function and structure, 36 untreated patients with essential hypertension (WHO class I and II) were examined. Posterior wall thickness, relative wall thickness, and left ventricular mass were determined by M-mode echocardiography. Plasma renin activity, aldosterone, angiotensin I, and angiotensin II levels were measured by radioimmunoassay. Plasma renin activity was related to 24-hour urinary sodium excretion. Of all the endocrine parameters, only the angiotensin II level correlated with posterior wall thickness (r = 0.50, p less than 0.05) and relative wall thickness (r = 0.46, p less than 0.05). This relationship was confirmed by stepwise multiple regression analysis taking arterial pressure, obesity, and sodium excretion into account (p less than 0.05). Plasma renin activity but not the angiotensin II level correlated positively with the ejection fraction (r = 0.42, p less than 0.05) and velocity of circumferential fiber shortening (r = 0.57, p less than 0.01). Thus, angiotensin II emerged as a determinant of left ventricular structural adaptation in essential hypertension. PMID- 2975464 TI - Cardiac pathophysiology and its heterogeneity in patients with established hypertensive disease. The first Robert C. Tarazi lecture. PMID- 2975465 TI - Coronary hemodynamics in hypertensive heart disease. Basic concepts, clinical consequences, and experimental analysis of regression of hypertensive microangiopathy. AB - Myocardial hypertrophy may influence coronary hemodynamics variably. Therefore, coronary sinus blood flow (gas chromatic argon technique) was determined in patients with left ventricular hypertrophy, with or without dilatation, associated with entirely normal coronary arteriographic results: 12 patients with hypertrophic obstructive cardiomyopathy (left ventricular mass-to-volume ratio, 3.66 +/- 0.52 g/ml), 22 patients with hypertensive heart disease due to essential hypertension (left ventricular mass-to-volume ratio, 2.12 +/- 0.26 g/ml), 18 patients with hypertensive dilatation (left ventricular mass-to-volume ratio, 1.6 +/- 0.48 g/ml), six patients with aortic stenosis (left ventricular mass-to volume ratio, 1.99 +/- 0.41 g/ml), 12 patients with aortic incompetence, and 20 patients with normal heart function. Coronary sinus blood flow was determined as a control value and as the value following intravenous injection of dipyridamole (0.5 mg/kg of body weight). Coronary reserve was calculated as the ratio of coronary resistance before and after dipyridamole. Normal coronary reserve averaged 4.89 +/- 0.11. Similar values, despite marked left ventricular hypertrophy, were present for both hypertrophic obstructive cardiomyopathy (4.4 +/- 0.19) and aortic stenosis (4.66 +/- 0.12), whereas coronary reserve was considerably reduced in the concentrically hypertrophied hypertensive hearts (3.22 +/- 0.19) (p less than 0.001). Moderate decrease in coronary reserve was found in aortic incompetence and in dilated essential hypertension. These results indicate that patients with nonhypertensive hypertrophy, despite left ventricular mass augmentation, may have normal coronary reserve, whereas at a comparable degree of left ventricular hypertrophy, patients with hypertensive hypertrophy have a specific reduction in coronary reserve. Independent from vascular effects, ventricular dilatation may result in deterioration of coronary reserve because of an abnormal component of coronary vascular resistance. These results were also verified in experimental hypertension. Moreover, prevention and/or regression of the impaired coronary circulation in experimental hypertensive heart disease, most probably due to the reduction of smooth muscle layers of the media of coronary resistance vessels, could be achieved by long-term vasodilator therapy. PMID- 2975466 TI - Cardiopulmonary receptor regulation of renin release. AB - Cardiopulmonary receptors have been shown to modulate renin release in animals. However, their involvement in reflex control of renin in humans has never been unequivocally established. This report reviews data on the effects on plasma renin activity of maneuvers (lower body negative pressure and passive leg raising) that reduce and increase central venous pressure and cardiac diameter without affecting blood pressure and heart rate, thereby deactivating and stimulating cardiopulmonary receptors with little or no involvement of the arterial baroreceptors. In normotensive subjects, reduction in central venous pressure was accompanied by an increase in plasma renin activity that was similar to the increase observed during tilt that reduced central venous pressure to a similar extent. Conversely, an increase in central venous pressure was accompanied by a reduction in plasma renin activity. The increase in plasma renin activity that followed the reduction in central venous pressure was drastically attenuated in four patients who had undergone cardiac transplantation, along with other reflex effects (changes in forearm vascular resistance and plasma norepinephrine) of cardiopulmonary receptor manipulations. There was a modest reduction in the overall reflex changes in plasma renin activity in patients with mild to moderate essential hypertension compared with normotensive subjects. However, patients with essential hypertension and cardiac hypertrophy showed marked attenuation of all reflex influences of the cardiopulmonary receptors. In five subjects, therapeutic regression of this structural alteration was associated with a clear improvement in the cardiopulmonary reflex. Thus, in humans, cardiopulmonary receptors exert an important reflex control of renin release. This control (which is due in part to receptors located in the cardiac walls) is moderately affected by a mild to moderate blood pressure elevation but is markedly impaired when the elevation produces structural alterations in the heart. Preliminary evidence, however, suggests that the cardiopulmonary reflex may be improved by a reduction in cardiac hypertrophy. PMID- 2975467 TI - Plasma levels of atrial natriuretic factor in inappropriate secretion of antidiuretic hormone syndrome. PMID- 2975468 TI - The evolution of occupational therapy in the public schools: the laws mandating practice. AB - This paper provides an overview of the evolution of occupational therapy in public school systems. The focus of this history is the passage and implementation of the laws mandating service provision to children with disabilities and a cursory examination of the social, political, and economic currents that have influenced that service provision. An effort was made to cull the most salient issues currently affecting occupational therapists who work in public school systems; for this reason, information was gathered from oral history sources, which provided the bulk of the data. Additional written sources were synthesized to provide a comprehensive review of the story of occupational therapy's emergence and current responsibilities in the education of children with disabilities. PMID- 2975469 TI - Special education and occupational therapy: making the relationship work. AB - Working as an occupational therapist in publicly funded schools requires a variety of skills. These skills include assessing the needs of children, serving as a member of the multidisciplinary team, developing individualized education program (IEP) goals and objectives in conjunction with other team members, providing services, and coordinating efforts with parents, teachers, and administrators. To fulfill these responsibilities, occupational therapists must have a comprehensive understanding of the complex federal and state laws that mandate the provision of special education and related services. Therefore, the purposes of this article are (a) to describe the legal framework within which decisions are made to provide occupational therapy to students in publicly funded school programs and (b) to highlight the knowledge and skills occupational therapists need to work effectively in schools with teachers, administrators, and parents. PMID- 2975470 TI - Promoting occupational therapy in the schools. AB - In response to our perceptions of the need to clarify the nature of school-based occupational therapy services at the local, state, and federal levels, we have written this article to address three areas. First, we suggest that the terminology used by school-based occupational therapists to describe their work be explained in terms that will allow parents and educators to better understand the roles and functions of the occupational therapist in the school. Second, to clarify and thereby increase the marketability of school-based occupational therapy, we propose a conceptual framework upon which the provision of occupational therapy as a service related to education can be based. Third, to clarify the unique role of occupational therapy in the schools, we define the roles and functions of school-based occupational therapy in such a way as to differentiate it from other related school-based services. Finally, we identify strategies to further secure the position of occupational therapy in the schools. PMID- 2975471 TI - The changing environment of early intervention services: implications for practice. AB - A new plan for providing early intervention services for very young children and their families has been established with Part H of the Education of the Handicapped Act Amendments of 1986, Public Law 99-457. Each state is currently exploring how it can best develop one comprehensive system of care that combines health, education, social, and family services. Health practitioners have traditionally provided early intervention services in medical settings such as hospitals and local health departments since the early 1900s. Educators, prompted by federal legislation, have provided early intervention services in the public schools primarily since 1975. Occupational therapy practitioners work in both medical and educational settings and must now meet the numerous challenges and opportunities in developing comprehensive, family-centered, community-based care for very young children with special needs. This paper traces the historical development of early intervention services and discusses the major practice issues associated with implementing Part H of the Education of the Handicapped Act Amendments of 1986. PMID- 2975472 TI - Pulmonary artery stenosis and balloon catheter angioplasty in the pulmonary circulation. A case report and review of the literature. PMID- 2975473 TI - Diagnosis and management of low back pain. PMID- 2975474 TI - Alopecia mucinosa progressing into mycosis fungoides. A long-term follow-up study of two patients. AB - The present report describes two patients with alopecia mucinosa who developed mycosis fungoides 4 and 8 years after the diagnosis of alopecia mucinosa had been made. The relationship between both conditions will be discussed. PMID- 2975475 TI - Reduced IgG Fc-receptor-mediated phagocytosis in human monocytes isolated from alcoholics. AB - Blood monocytes (MC) were isolated from alcoholics and from age-matched controls. Phagocytosis mediated by the IgG Fc-receptor (Fc-R) was assessed in the MC immediately after isolation by using IgG-opsonized sheep erythrocytes as test particles. Experiments were also performed after preincubating the MC in medium containing 10% autologous serum with or without ethanol (80 mM) in vitro at 37 degrees C for 15 min, and with or without ethanol (80 mM) present during the internalization assay in vitro (37 degrees C for 60 min). MC from alcoholics exhibited lower phagocytic capacity via the Fc-R than MC from healthy controls, approximately 50% versus 70% phagocytosing cells. Preincubation of the MC in medium containing 10% autologous serum with 80 mM ethanol, reduced the percentage of phagocytosing MC from controls to 55%, whereas no further reduction occurred in cells from alcoholics. When present during the assays of Fc-R function, ethanol decreased MC phagocytosis of IgG-coated particles significantly in both test groups, to about 30% phagocytosing cells. Blood samples disclosed increased serum levels of markers of liver dysfunction, immunoglobulins, increased sedimentation rate, and white blood cell counts among the alcoholics, while their serum levels of 1,25-dihydroxyvitamin D3 were reduced. PMID- 2975476 TI - Interaction of stress and ethanol: effect on beta-endorphin and catecholamines. AB - To examine the interaction of ethanol (ET) and stress on beta-endorphin and catecholamine (CA) levels, male rats pretreated with ET (3.0 g/kg, i.p.) or saline were immobilized for 30 min and killed 90 min after the initial injection. Stress resulted in (a) an increase in plasma levels of norepinephrine (NE, 243%), epinephrine (E, 175%), beta-endorphin (220%) and corticosterone (CS, 151%) and a decrease in dopamine (DA, 54%); (b) a decrease in hypothalamic NE (15%) and beta endorphin (33%) levels and an increase E (23%) and DA (58%) levels; (c) a decrease in pituitary beta-endorphin levels in both the neurointermediate (23%) and anterior (131%) lobes. Treatment with ET resulted in: (a) an increase in plasma NE (81%), E (53%), CS (71%), and beta-endorphin (33%) levels and decrease in DA (54%); (b) a decrease in the hypothalamic NE (12%) levels and an increase DA (27%) and beta-endorphin (46%) levels, and (c) a decrease in beta-endorphin (15.5%) in the intermediate lobe of the pituitary. Treatment with ET of stressed animals had only a small effect: (a) in plasma NE, E, CS, and beta-endorphin levels decreased by 30, 31, 14, and 36%, respectively; (b) in the hypothalamus DA levels decreased by 40% and beta-endorphin increased by 71%; (c) in the pituitary beta-endorphin increased in both the intermediate lobe (25%) and anterior (50%) lobes. Thus when the data of the stressed ET-treated group is compared to that of the nonstressed saline injected group, none of the measures differ significantly. These results confirm our earlier work indicating a significant interaction of ET and stress. PMID- 2975477 TI - Quantification of quinolinic acid in rat brain, whole blood, and plasma by gas chromatography and negative chemical ionization mass spectrometry: effects of systemic L-tryptophan administration on brain and blood quinolinic acid concentrations. AB - A gas chromatography/mass spectrometry assay is described to quantify the endogenous neurotoxin quinolinic acid (QUIN) in brain, whole blood, and plasma. High specificity and high sensitivity were obtained by using negative chemical ionization and accuracy was achieved by using [18O]QUIN as internal standard. Neutralized perchloric acid extracts were washed with chloroform, applied to Dowex 1 x 8 (formate form), and eluted with 6 M formic acid. After lyophilization, QUIN and [18O]QUIN were esterified with hexafluoroisopropanol (to mass 467 and 471, respectively) using trifluoroacetylimidazole as catalyst. The esters were extracted into heptane and injected onto a gas chromatograph, DB-5 capillary column. QUIN and [18O]QUIN were quantified by selected ion monitoring of QUIN-specific anion currents from the molecular anions (m/z 467 and 471, respectively) and a specific anion fragment (m/z 316 from QUIN and m/z 320 from [18O]QUIN). Minimum sensitivity was 3 fmol, intraassay variability was 3.2%, and interassay variability was 8.1% QUIN concentrations in frontal cortex from over 200 rats ranged from 20 to 180 fmol/mg wet wt. Two hours after systemic L tryptophan (L-Trp; 0.370 mmol/kg) administration, QUIN increased in whole blood 134.8-fold and in plasma, 74.3-fold. In frontal cortex, increases in QUIN (22.6 fold, corrected for QUIN in blood) exceeded increases in cortical L-Trp (2.54 fold), 5-HT (1.35-fold), and 5-HIAA (1.74-fold). These studies demonstrate that QUIN is present in brain and is sensitive to the availability of systemic L-Trp. PMID- 2975478 TI - [Percutaneous intraluminal angioplasty in renal artery stenoses in a kidney considered to be solitary (solitary kidney, remaining kidney, before contralateral nephrectomy)]. PMID- 2975479 TI - The significance of estrogen receptors in tamoxifen and toremifene therapy. AB - Estrogen receptor (ER) concentration of breast cancer tissue is important in predicting the response of each patient to hormonal, especially antiestrogen treatment. About half of the patients with ER rich tumours respond and only about 10% of the patients with ER poor tumours respond to antioestrogen treatment. Tamoxifen is a well known and widely used drug. Toremifene is a new antioestrogen, developed in Finland. At standard doses both compounds have comparable hormonal and antitumour effects, and there is no clear difference between the compounds in the affinity to ER. The value of ER in predicting the response to tamoxifen and toremifene therapy in ER positive breast cancer is significant. It is not known, however, if the role of ER remains the same with high dose toremifene. Although ERs are an important predictive factor, the antioestrogens evidently act through them only in part. As the prediction is correct in about half of the patients, other mechanisms must influence tumour growth regulation, such as the expression of oncogenes and the synthesis and activity of growth factors. PMID- 2975480 TI - [Diagnostic and therapeutic aspects of peritoneal tuberculosis in Morocco. Apropos of 300 cases]. AB - We describe 300 cases of peritoneal tuberculosis observed in the space of 11 years in Morocco. The most usual terrain presented is the young woman in precarious socio-economic conditions. The febrile ascitic form was observed in 229 cases. The ulcero-caseous was observed in 51 cases, and the fibro-adhesive form in 14 cases. Diagnostic was based on laparoscopy with peritoneal biopsy. A controlled prospective study done randomly on 100 patients showed that corticotherapy associated with tuberculous antibiotherapy did not speed up clinical recovery but tended, rather, to slow down the disappearance of granulations and indeed facilitate the constitution of peritoneal adherence phenomena. PMID- 2975481 TI - Role of macrophages in cytotoxic reaction against cells of lymphatic leukaemia L1210 in mice in the light of cytoenzymatic and ultracytoenzymatic studies. PMID- 2975483 TI - The dentist's contribution to rehabilitation of cervical posture and function: orthopedic and neurological considerations in the treatment of craniomandibular disorders. PMID- 2975482 TI - Effects of antiandrogen-estrogen treatment on sexual and endocrine parameters in hirsute women. AB - Sexual activity was evaluated in 51 women with hirsutism associated with increased levels of circulating androgens before and while on combined treatment with the antiandrogen cyproterone acetate (CA) and ethinyl estradiol (EE2) and compared to a reference group of 52 subjects. The percentage of unbound testosterone (T) was higher (p less than 0.001), the coital frequency lower (p less than 0.05), and the masturbation frequency higher (p less than 0.04) in hirsute women. Mean frequency of total activity (coitus plus masturbation) was similar in the two groups. Treatment with combination of CA and EE2 resulted in a decline of unbound T (p less than 0.001). There was no change of total sexual activity, but coital frequency increased (p less than 0.05) and masturbation frequency declined (p less than 0.04). It is concluded that raised levels of circulating androgens, as judged by free T concentration, are not of crucial importance in the expression of sexual behavior in hirsute women. PMID- 2975484 TI - TMJ syndrome: dysfunction & chronic pain. PMID- 2975485 TI - Dentistry's role is changing. PMID- 2975487 TI - Craniomandibular disorders, neurophysiology, and auriculotherapy. PMID- 2975486 TI - The American College of Orthopaedic Medicine. PMID- 2975488 TI - Nervous system. PMID- 2975489 TI - Mouthguards: protection against shock to head, neck and teeth. PMID- 2975490 TI - The dental distress syndrome quantified. PMID- 2975491 TI - Hepatitis B immunization in high risk neonates born from HBsAg and HBeAg positive mothers: comparison of standard and low dose regimens. AB - A reduced dose of plasma derived hepatitis B vaccine (Hevac B) was tested for efficacy in the prevention of perinatal hepatitis B virus (HBV) transmission in high risk neonates born from e-antigen positive HBsAg carrier mothers. Forty newborn infants born of these mothers were given hepatitis B immune globulin (HBIG) 100 IU intramuscularly immediately after birth, combined with either standard or reduced doses of HBV vaccine. The infants were divided into two groups of 20 infants each. The standard dose of HBV vaccine (5 micrograms) was given to group I, while infants in group II received reduced dose (2 micrograms) at birth and at 1, 2 and 12 months of age. There was no statistically significant difference in the efficacy and antibody responses of these two combined prophylaxis regimens. The protective efficacy rate of HBV vaccine was found to be 94.0 and 93.2 percent in group I and group II, respectively. At twelve months of age, the anti-HBs seroconversion rates were 80.0 percent in group I and 86.7 percent in group II, with geometric mean titres of 84.57 mlU/ml and 78.56 mlU/ml, in group I and group II, respectively. One month after a booster at one year of age, anti-HBs could be detected in 86.7 percent of the infants in both groups. The geometric mean titres were 429.04 and 664.81 mlU/ml, in group I and group II, respectively. Anti-PreS2 antibody was detected in high titre as early as 4 months after the first dose of HBV vaccine, with a geometric mean titre of 116.30 mlU/ml and 107.97 mlU/ml, in group I and group II, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975492 TI - Effect of different treatment regimens with ethanol and p-chlorophenylalanine on tolerance development in UChA and UChB rats. AB - Since we have previously observed a genetic difference in the development of tolerance to the narcotic effect of ethanol in UChA and UChB rats when providing them with a 10% v/v ethanol solution as sole drinking fluid, experiments were performed in order to know whether the resistance to development of tolerance to ethanol in UChB rats was also exhibited after other regimens of ethanol administration, namely, a 2.76 g/kg ethanol IP injection 24 hr before the experiment, only 10% v/v ethanol solution as sole drinking fluid for 21 days, or receiving acutely a daily dose of 2.76 g/kg ethanol by gavage for seven days. Participation of serotoninergic neurons was tested by treating rats with p chlorophenylalanine (p-CPA), a known serotonin depletor. Results show that UChA rats developed tolerance to ethanol-induced narcosis and hypothermia, while UChB rats developed it to narcosis only when they received acute oral doses of ethanol for 7 days and did not develop tolerance to hypothermia with any of the treatment regimens. p-CPA pretreatment did prevent the development of tolerance in both strains of rats, confirming the participation of serotoninergic neurons in ethanol tolerance in rats. PMID- 2975493 TI - Infectious complications in patients with HIV infection. PMID- 2975494 TI - Regional capillary and myocyte distribution in normal and exercise trained male and female rat hearts. AB - Adult Sprague-Dawley male and female rats were exercise trained for five months by either treadmill running or swimming. Significant differences in left ventricular regional capillary density and myocyte cross-sectional area were found. In control rats the epicardial regions had greater capillary density than endocardial regions. Endocardial myocyte cross-sectional areas were greater than those of epicardial myocytes in both sexes. Male rats had larger endocardial myocytes and larger hearts than females. After exercise training, myocyte size increased in the epicardial region but not in the endocardial region, while capillary density increased significantly only in the endocardial region. Similar changes were seen in both male and female rats with comparable degrees of exercise induced hypertrophy. These data suggest that exercise training "normalizes" the distribution of capillaries in the myocardium. Capillary density increased only in the regions where myocyte cross-sectional area did not increase. Further, the effects of exercise on male and female rat hearts is not different when the degree of exercise induced hypertrophy is similar. PMID- 2975495 TI - [Analysis of tumor specific immunological response in the patients with malignant brain tumors by the mixed lymphocyte-tumor cell reaction]. AB - In considering immunological approaches to treatment of the patients with malignant brain tumors, it seems very important to enhance the tumor specific immunity. Then, to ascertain whether tumor specific immune response occurs in these patients with malignant brain tumors, lymphocyte blastogenetic responses to tumor cells were examined in 18 patients with malignant brain tumors. Furthermore, to compare the systemic immunological responses with the local responses in the brain tumor tissues, both peripheral blood lymphocyte (PBL) and tumor infiltrating lymphocyte (TIL) were used as blastogenetic stimulators to the tumor cells. The PBL from the patients with malignant gliomas showed any positive blastogenetic response to their own glioma cells in 7 or 12 cases (about 58%). But, TIL from these patients showed a positive response in the only 3 cases (25%). In 6 cases of metastatic brain tumors, otherwise, their PBL showed any positive blastogenetic response to their own tumor cells in 4 of 6 cases (about 67%), and their TIL showed any positive blastogenetic responses in these same 4 cases. So, the tumor specific immunological responses may be stronger in the patient with metastatic brain tumors than in the patient with malignant gliomas. These immunological responses were, furthermore, more weak in the brain tumor tissues than in the systemic immunity. Then, this lymphocyte blastogenetic response to tumor cells were compared with other lymphocyte stimulating examination such as rectine or allogenetic lymphocyte stimulation. Our studies revealed that this lymphocyte blastogenetic response to tumor cells were at lower level compared with rectine such as PHA, PWM, and Con A, or allogenetic lymphocyte stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975496 TI - Controlled preclotting--a simplified technique proved by time. AB - Precise preclotting techniques are necessary to ensure that knitted Dacron (DuPont, Inc., Wilmington, DE) grafts are impervious and hypothrombogenic. In situ and submersion preclotting techniques are insufficient because they result in a highly thrombogenic flow surface and may not produce an impervious wall, a bad combination at best and a disastrous one at worst. The authors have used a controllable preclotting method for more than 10 years that assures that the wall will be impervious and the flow surface hypothrombogenic. We believe that the 100 200 ml of the patient's blood and approximately 15 min of the surgeon's time required to achieve this result are prudent investments for both. PMID- 2975497 TI - Peritoneal catheter development. Currently used catheters- advantages/disadvantages/complications, and catheter tunnel morphology in humans. AB - Original Tenckhoff catheters are still the most commonly used catheters, and catheter survival rates are still unsatisfactory. Preliminary results with Swan Neck catheters, the catheters with a permanently bent intramural segment, are encouraging since these catheters are associated with low complication rates. The role of the external cuff in prevention of exit and tunnel infections is still controversial. Animal experiments indicate that collagen ingrowth into the external cuff is crucial in the inhibition of epidermal downgrowth. The author's recent preliminary observations on four catheter tunnels removed 1 month to 5.5 years after catheter implantations, indicate that 1) epidermal cells in humans do not spread deeper than a few millimeters from the skin exit and 2) the external cuff does not participate in the epidermal cell downgrowth inhibition. This is unlike animal studies. The difference in epidermal biology between humans and animals may render animal experiments difficult to apply to catheter technology in humans. PMID- 2975498 TI - The peritoneoscopic implantation of a polyurethane percutaneous access device for peritoneal dialysis. Preliminary experience. AB - The short-term experience with peritoneoscopic implantation of eight Dermaport devices is discussed. These eight devices were implanted using local anesthesia and utilizing a modification of the Y-TEC method of catheter implantation. Excellent immediate function was observed universally allowing catheter use for dialysis and good skin-device interface; however dialysate leakage occurred within 2 to 4 weeks in four patients. Lack of permanent bonding between the catheter and the adjustable proximal cuff, resulting in suboptimal or slow ingrowth of tissue around it, appears to be the cause of this phenomenon. PMID- 2975499 TI - Therapeutic ratio of photodynamic therapy in the treatment of superficial tumours of skin and subcutaneous tissues in man. AB - Six patients with a total of 34 assessible subcutaneous or cutaneous lesions were treated with photodynamic therapy using 1.0, 1.5 or 2.0 mg kg-1 of photofrin II and 25-100 J cm-2 of red light (630 nm). The incidence of complete tumour response and skin necrosis were used to try to assess the therapeutic ratio of photodynamic therapy. The tumour response rate was 47%. The rate of tumour control and necrosis increased in parallel with dose of photosensitizer and light used, implying a low therapeutic ratio. However, the use of necrosis with eschar formation as an end-point for severe normal tissue damage is questioned as the skin healed completely in all cases and with minimal discomfort to the patients. PMID- 2975500 TI - The mixed epidermal cell lymphocyte-reaction is the most predictive factor of acute graft-versus-host disease in bone marrow graft recipients. AB - Risk factors for acute graft-versus-host disease (GvHD) remain controversial. We performed uni- and multivariate statistical analyses on a series of 37 patients receiving a non-depleted allogeneic bone marrow transplant from an HLA-identical sibling donor for a haematological malignancy, in order to identify risk factors for GvHD. Three factors were associated with development of moderate to severe GvHD: a positive mixed epidermal cell-lymphocyte reaction (MECLR) between donor and recipient, previous pregnancies in female donors and chronic myeloid leukaemia diagnosis. The MECLR was the most important predictive factor, selected in first rank by the stepwise linear discriminant analysis. Combining these three prognostic factors in the jackknifed procedure, we could correctly classify 33/37 patients in two groups: grade O-I versus grade II-IV acute GvHD. These results should apply to donor selection and to predict donor/recipient pairs at high risk of GvHD who might benefit of bone marrow T-cell depletion and those at low risk for whom depletion could be avoided. PMID- 2975501 TI - A streptokinase dependent plasma factor (SKDF) induces leucocyte tissue factor activity. AB - Various inducers endow human leucocytes with a procoagulant activity of tissue factor type. We have observed a novel plasma factor which in combination with streptokinase induces powerful leucocyte procoagulant activity. This streptokinase dependent factor (SKDF) is present in normal plasma or serum albeit quantitatively different in individual donors. The generation of tissue factor activity as a function of streptokinase-plasma complex shows a specific and saturable sigmoidal dose-response curve. The Hill plot shows a straight line with Hill coefficient, H = 2.2, suggesting a strong positive cooperativity for the binding of this streptokinase-plasma complex to the leucocyte surface receptor for the signal transduction leading to the biosynthesis of tissue factor apoprotein. It also suggests that the leucocyte surface receptor for streptokinase-plasma complex differs from that for endotoxin lipopolysaccharides. SKDF is of apparent high molecular weight. It does not appear to be an antibody to streptokinase since its level does not correlate with the level of antibodies to streptokinase, and it does not correlate with the antistreptolysin titre. Furthermore, SKDF does not bind to protein A. It has a narrow pH range of stability, and is destroyed at 56 degrees C, or at freeze-drying, Urokinase, another plasminogen activator, or plasmin were unable to activate SKDF to induce the leucocyte procoagulant activity. SKDF may play a role in thrombolytic therapy. PMID- 2975502 TI - Replacement therapy for a homozygous protein C deficiency-state using a concentrate of human protein C and S. AB - A severe congenital deficiency of protein C was diagnosed in a 10-month-old girl who had been suffering from skin necrosis since the age of 7 months. The patient was treated initially with fresh frozen plasma, 10 ml per kg body weight, every 24 h. Following treatment, the mean plasma level of protein C was 0.1 U/ml after 30 min and less than 0.02 U/ml after 24 h. The child was then treated with a concentrate of human protein C and S, 100 U protein C per kg body weight, given every 48 h for a period of 9 months. The mean plasma level of protein C was 0.93 U/ml 30 min after administration of the concentrate and 0.13 and 0.08 U/ml after 24 and 48 h, respectively. The mean post-transfusional in vivo recovery of protein C was 44% and the half life was 8.3 h. The mean plasma level of 'free' protein S increased from 1.1 to 2.2 U/ml after administration of the concentrate. There was no increase in 'bound' protein S. The in vivo recovery of 'free' protein S was 49% and the half life was about 17 h. Since the start of this replacement therapy using a human protein C and S concentrate, the patient has not developed any thromboembolic complications. These results indicate the therapeutic value of human protein C and S concentrate in the treatment of severe protein C deficiency. PMID- 2975503 TI - Risk of recurrence of occupational back pain over three year follow up. AB - A random sample including 2342 cases representative of all occupational back injuries in Quebec (1981) was followed up prospectively over three years to assess the recurrence rate of back problems (lumbar, thoracic, and cervical). Each medical and accident report was reviewed to obtain the site of symptoms and occupation. Age, sex, industrial sector, and number of episodes of absence from work were abstracted from the computerised Quebec Compensation Board files. The recurrence rate was 20.0% at one year follow up and 36.3% at three years. A multivariate analysis using a Poisson regression, was performed to model the risk of recurrence over time. Men had a higher chance of recurrence (risk ratio = 1.85, 95% CI = 1.50-2.27) but among recurrent cases, the average total number of episodes was comparable between men and women. Age showed a protective effect on the probability of recurrence (10 years: RR = 0.93, 95% CI = 0.88-0.98) due to the lower recurrence rate in the 45-64 year old group (31.8%). Cervical and lumbar symptoms had identical recurrence profiles whereas thoracic symptoms had a significantly lower recurrence rate. Drivers had the highest recurrence rate (42.1%) and nurses had the highest average number of recurrences (2.03) among recurrent cases. Both occupations had statistically significant excesses after controlling for the other variables. PMID- 2975504 TI - Ultralow volume application of organophosphate concentrate in grain terminals: a new occupational health hazard. AB - The introduction of ultralow volume (ULV) application of the organophosphate pesticide Fenitrothion in grain terminals presents a risk to workers of skin contact with concentrate. Blood testing, by the Ellman method, of a group of five grain terminal workers working on grain treatment showed a lowering of mean red blood cell cholinesterase (RBC ChE) activity to 23 units/gm Hb (normal value 28 40) with a range of 16-29. The probable cause was identified as percutaneous absorption of Fenitrothion concentrate by workers using ungloved hands to clean blocked drip feed nozzles. Modification of work practices was followed by a rise of mean RBC ChE to 33.6 units/gm Hb (range 32-36) during the following grain treatment season. RBC ChE activity measured during the intervening winter season- that is, a non-exposure period--showed a mean of 33.3 units/gm Hb (range 23-40). PMID- 2975505 TI - Filament assembly and regulation of the actin-activated ATPase activity of thymus myosin. AB - The effects of light chain phosphorylation on the actin-activated ATPase activity and filament assembly of calf thymus cytoplasmic myosin were examined under a variety of conditions. When unphosphorylated and phosphorylated thymus myosins were monomeric, their MgATPase activities were not activated or only very slightly activated by actin, but when they were filamentous, their MgATPase activities were stimulated by actin. The phosphorylated myosin remained filamentous at lower Mg2+ concentrations and higher KC1 concentrations than did the unphosphorylated myosin, and the myosin concentration required for filament assembly was lower for phosphorylated myosin than for unphosphorylated myosin. By varying the myosin concentration, it was possible to have under the same assay conditions mostly monomeric myosin or mostly filamentous myosin; under these conditions, the actin-activated ATPase activities of the filamentous myosins were much greater than those of the monomeric myosins. The addition of phosphorylated myosin to unphosphorylated myosin promoted the assembly of unphosphorylated myosin into filaments. These results suggest that phosphorylation may regulate the actomyosin-based motile activities in vertebrate nonmuscle cells by regulating myosin filament assembly. PMID- 2975506 TI - Placental anticoagulant proteins: isolation and comparative characterization four members of the lipocortin family. AB - Previously we isolated and characterized a placental anticoagulant protein (PAP or PAP-I), which is a Ca2+-dependent phospholipid binding protein [Funakoshi et al. (1987) Biochemistry 26, 5572] and a member of the lipocortin family [Funakoshi et al. (1987) Biochemistry 26, 8087]. In this study, three additional anticoagulant proteins (PAP-II, PAP-III, and PAP-IV) were simultaneously isolated from human placental homogenates prepared in the presence of 5 mM ethylenediaminetetraacetic acid. The isoelectric points of PAP-I, PAP-II, PAP III, and PAP-IV were 4.8, 6.1, 5.9, and 8.1, respectively, and their apparent molecular weights were 32,000, 33,000, 34,000, and 34,500, respectively. Amino acid sequences of cyanogen bromide fragments of these proteins showed that PAP III was a previously unrecognized member of the lipocortin family, while PAP-II was probably the human homologue of porcine protein II and PAP-IV was a derivative of lipocortin II truncated near the amino terminus. Comparative studies showed that all four proteins inhibited blood clotting and phospholipase A2 activity with potencies consistent with their measured relative affinities for anionic phospholipid vesicles. However, PAP-IV bound to phospholipid vesicles approximately 160-fold more weakly than PAP-I, while PAP-II and PAP-III bound only 2-fold and 3-fold more weakly. These results increase to six the number of lipocortin-like proteins known to exist in human placenta. The observed differences in phospholipid binding may indicate functional differences among the members of the lipocortin family despite their considerable structural similarities. PMID- 2975507 TI - Drosophila acetylcholinesterase: demonstration of a glycoinositol phospholipid anchor and an endogenous proteolytic cleavage. AB - The presence of a glycoinositol phospholipid anchor in Drosophila acetylcholinesterase (AChE) was shown by several criteria. Chemical analysis of highly purified Drosophila AChE demonstrated approximately one residue of inositol per enzyme subunit. Selective cleavage by Staphylococcus aureus phosphatidylinositol-specific phospholipase C (PI-PLC) was tested with Drosophila AChE radiolabeled by the photoactivatable affinity probe 3-(trifluoromethyl)-3-(m [125I]iodophenyl)diazirine [( 125I]TID), a reagent that specifically labels the lipid moiety of glycoinositol phospholipid-anchored proteins. Digestion with PI PLC released 75% of this radiolabel from the protein. Gel electrophoresis of Drosophila AChE in sodium dodecyl sulfate indicated prominent 55- and 16-kDa bands and a faint 70-kDa band. The [125I]TID label was localized on the 55-kDa fragment, suggesting that this fragment is the C-terminal portion of the protein. In support of this conclusion, a sensitive microsequencing procedure that involved manual Edman degradation combined with radiomethylation was used to determine residues 2-5 of the 16-kDa fragment. Comparison with the Drosophila AChE cDNA sequence [Hall, L.M.C., & Spierer, P. (1986) EMBO J. 5, 2949-2954] confirmed that the 16-kDa fragment includes the N-terminus of AChE. Furthermore, the position of the N-terminal amino acid of the mature Drosophila AChE is closely homologous to that of Torpedo AChE. The presence of radiomethylatable ethanolamine in both 16- and 55-kDa fragments was also confirmed. Thus, Drosophila AChE may include a second posttranslational modification involving ethanolamine. PMID- 2975508 TI - 9,13-dicis-rhodopsin and its one-photon-one-double-bond isomerization. AB - Incubation of purified 9,13-dicis-retinal with cattle opsin in 2% digitonin at 20 degrees C produced two pigments, one unstable (lambda max 478 nm) and the other stable (lambda max 485 nm) in hydroxylamine. The two pigments exhibited different characteristics. HPLC analysis revealed that the chromophores of these pigments have respectively 9,13-dicis and 9-cis geometries. Under various conditions the amount of 9,13-dicis-rhodopsin formed never exceeded 30% of the total pigments. The addition of 9,13-dicis-retinal to the ROS suspension containing opsin produced 9-cis-rhodopsin in 97% yield. Irradiation of the 9,13-dicis-rhodopsin mainly produced 13-cis-retinal, while 9-cis-rhodopsin produced the all-trans isomer. These results demonstrated that the one-photon-one-double-bond isomerization process took place in 9,13-dicis-rhodopsin. PMID- 2975509 TI - Metabolism of D-glucose in a wall-less mutant of Neurospora crassa examined by 13C and 31P nuclear magnetic resonances: effects of insulin. AB - 13C NMR and 31P NMR have been used to investigate the metabolism of glucose by a wall-less strain of Neurospora crassa (slime), grown in a supplemented nutritionally defined medium and harvested in the early stationary stage of growth. With D-[1-13C]- or D-[6-13C]glucose as substrates, the major metabolic products identified from 13C NMR spectra were [2-13C]ethanol, [3-13C]alanine, and C1- and C6-labeled trehalose. Several observations suggested the existence of a substantial hexose monophosphate (HMP) shunt: (i) a 70% greater yield of ethanol from C6- than from C1-labeled glucose; (ii) C1-labeled glucose yielded 19% C6 labeled trehalose, while C6-labeled glucose yielded only 4% C1-labeled trehalose; (iii) a substantial transfer of 13C from C2-labeled glucose to the C2-position of ethanol. 31P NMR spectra showed millimolar levels of intracellular inorganic phosphate (Pi), phosphodiesters, and diphosphates including sugar diphosphates and polyphosphate. Addition of glucose resulted in a decrease in cytoplasmic Pi and an increase in sugar monophosphates, which continued for at least 30 min. Phosphate resonances corresponding to metabolic intermediates of both the glycolytic and HMP pathways were identified in cell extracts. Addition of insulin (100 nM) with the glucose had the following effects relative to glucose alone: (i) a 24% increase (P less than 0.01) in the rate of ethanol production; (ii) a 38% increase (P less than 0.05) in the rate of alanine production; (iii) a 27% increase (P less than 0.05) in the rate of glucose disappearance. Insulin thus increases the rates of production of ethanol and alanine in these cells, in addition to increasing production of CO2 and glycogen, as previously shown. PMID- 2975510 TI - Binding of Ca2+ to the calcium adenosinetriphosphatase of sarcoplasmic reticulum. AB - The binding of Ca2+ and the resulting change in catalytic specificity that allows phosphorylation of the calcium ATPase of sarcoplasmic reticulum by ATP were examined by measuring the amount of phosphoenzyme formation from [32P]ATP, or 45Ca incorporation into vesicles, after the simultaneous addition of ATP and EGTA at different times after mixing enzyme and Ca2+ (25 degrees C, pH 7.0, 5 mM MgSO4, 0.1 M KCl). A "burst" of calcium binding in the presence of high [Ca2+] gives approximately 12% phosphorylation and internalization of two Ca2+ at very short times after the addition of Ca2+ with this assay. This shows that calcium binding sites are available on the cytoplasmic-facing side of the free enzyme. Calcium binding to these sites induces the formation of cE.Ca2, the stable high affinity form of the enzyme, with k = 40 s-1 at saturating [Ca2+] and a half maximal rate at approximately 20 microM Ca2+ (from Kdiss = 7.4 X 10(-7) M for Ca.EGTA). The formation of cE.Ca2 through a "high-affinity" pathway can be described by the scheme E 1 in equilibrium cE.Ca1 2 in equilibrium cE.Ca2, with k1 = 3 X 10(6) M-1 s-1, k2 = 4.3 X 10(7) M-1 s-1, k-1 = 30 s-1, k-2 = 60 s-1, K1 = 9 X 10(-6) M, and K2 = 1.4 X 10(-6) M. The approach to equilibrium from E and 3.2 microM Ca2+ follows kobsd = kf + kr = 18 s-1 and gives kf = kr = 9 s-1. The rate of exchange of 45Ca into the inner position of cE.Ca2 shows an induction period and is not faster than the approach to equilibrium starting with E and 45Ca. The dissociation of 45Ca from the inner position of cE.45Ca.Ca in the presence of 3.2 microM Ca2+ occurs with a rate constant of 7 s-1. These results are inconsistent with a slow conformational change of free E to give cE, followed by rapid binding-dissociation of Ca2+. PMID- 2975512 TI - Effect of endosulfan on adenosine triphosphatase (ATPase) activity in liver, kidney, and muscles of Channa gachua. PMID- 2975511 TI - [3H]spiperone binding to lymphocyte in extrapyramidal disease and in aging. AB - [3H]Spiperone binding to lymphocytes in Parkinson's disease (PD), Wilson's disease (WD), and age-matched control groups was studied. In the untreated PD group, [3H]spiperone binding was lower than in young controls, but did not differ from elderly healthy persons. After treatment with levodopa, the number of [3H]spiperone binding sites increased. In WD, lower binding of [3H]spiperone compared with age-matched controls was found. However, the magnitude of the differences in [3H]spiperone binding to lymphocytes in PD was too small to permit its use as a routine indicator of the disease state or the adequacy of pharmacological treatment in individual patients. [3H]Spiperone binding to lymphocytes decreases with age. Changes in [3H]spiperone binding to lymphocytes may be a general phenomenon for all states where dopamine is depleted, including normal aging. The nature of [3H]spiperone binding to lymphocytes remains unclear. The possible influence of dopamine on immune reactivity is discussed. PMID- 2975513 TI - In vitro effect of mercury and cadmium on brain Ca2+-ATPase of the catfish Ictalurus punctatus. PMID- 2975515 TI - [On the facial features of Down's syndrome as noted by Moire topography. 2. On the part proportionate to maxilla]. PMID- 2975514 TI - The use of dichloromethylene bisphosphonate and aminobutane bisphosphonate in hypercalcemia of malignancy. AB - 54 patients with malignant hypercalcemia were treated with either dichloromethylene bisphosphonate (Cl2MBP) or aminobutane bisphosphonate (AHButBP). Both compounds infused intravenously (i.v.) were rapidly effective in lowering serum calcium to normal range and a potency ratio between Cl2MBP and AHButBP could be roughly estimated as 1:100-250. 600 mg of Cl2MBP infused in one day over 9 h in nine patients were only slightly less effective (serum calcium from 12.3 +/- 1.7 to 10.1 +/- 1.1 SE, mg/dl) than 3300 mg infused in 11 consecutive days (13.4 +/- 1.9 to 9.3 +/- 1.2 SE, mg/dl). Thus the effectiveness of treatment with Cl2MBP was related better to the overall time of infusion than to the cumulative dose administered. This could not be demonstrated in a similar trial using AHButBP. 1600 mg Cl2MBP daily p.o. was unable to prevent the relapse of hypercalcemia in four patients after a treatment course with i.v. bisphosphonates. 100 mg of Cl2MBP intramuscularly maintained serum calcium within the normal range in two out of eight patients; however, weekly infusions of either Cl2MBP or AHButBP prevented the relapse of hypercalcemia in eight similarly treated patients for several weeks. Providing that adequate dosages are adopted, parenteral Cl2MBP and AHButBP may be a useful measure for controlling hypercalcemia in patients with carcinoma. PMID- 2975516 TI - [Studies on the glycolytic metabolism in Tawa sarcoma cells: inhibition of phosphofructokinase activity from Tawa sarcoma cells by nucleotide triphosphate]. PMID- 2975517 TI - [Light microscopical, electron microscopical and histochemical studies on the submandibular glands of Streptozotocin-induced rats]. PMID- 2975518 TI - [Tooth extraction in a patient who acquired artificial valve replacement]. PMID- 2975519 TI - [Facial features of Down's syndrome as noted by Moire topography. 3. On the feature of the facial outline]. PMID- 2975520 TI - Laser therapy in the treatment of cardiovascular disease. AB - Percutaneous laser angioplasty has now become a clinical reality, consisting chiefly of applications of thermal angioplasty in conjunction with balloon angioplasty for the recanalization of peripheral vascular obstructions. In conjunction with this development, various aspects of laser-tissue interactions, fibreoptic transmission and delivery catheter design pertinent to the cardiovascular system have come under closer scrutiny, resulting in the emergence of both noteworthy concepts and clinical achievements. PMID- 2975521 TI - Regional haemodynamic effects of depressor neuropeptides in conscious, unrestrained, Long Evans and Brattleboro rats. AB - 1. The regional haemodynamic effects of i.v. bolus doses of atrial natriuretic peptide (ANP, 1 and 10 nmol), calcitonin gene-related peptide (CGRP, 0.05 and 0.5 nmol) and corticotropin-releasing factor (CRF, 1 and 5 nmol) were assessed in conscious Long Evans and Brattleboro rats chronically instrumented with miniaturized, pulsed Doppler probes. 2. The low dose of ANP was without effect on mean arterial pressure (MAP), but caused tachycardia and hindquarters vasodilatation with vasoconstriction in renal and mesenteric beds in Long Evans rats. With the high doses of ANP these effects were more pronounced and MAP fell. In Brattleboro rats there was a primary renal vasodilatation. 3. The low dose of CGRP caused a slight fall in MAP in Long Evans rats, tachycardia and a renal vasodilatation. The high dose of CGRP caused marked hypotension, tachycardia and renal, mesenteric and hindquarters vasodilatation in both strains of rat. However, only in Long Evans rats were there secondary renal and mesenteric vasoconstrictions. 4. The low dose of CRF caused falls in MAP in both strains of rat, accompanied by renal and, particularly, mesenteric vasodilatation. Administration of the high dose of CRF caused profound, prolonged hypotension, tachycardia and mesenteric vasodilatation. There was also (late onset) hindquarters vasodilatation accompanying renal vasoconstriction that followed the initial vasodilatation in this vascular bed. 5. These results indicate that appropriate doses of particular peptides may be capable of promoting flow through individual peripheral vascular beds. PMID- 2975522 TI - Sodium-dependent inhibition by PN200-110 enantiomers of nicotinic adrenal catecholamine release. AB - 1. Dimethylphenylpiperazinium (DMPP) or high K concentrations evoke catecholamine release from perfused cat adrenal glands; in both cases the secretory response was significantly enhanced in the absence of Na. Tetrodotoxin did not modify the nicotinic secretory response. 2. The (+)- and (-)-enantiomers of the dihydropyridine Ca channel blocker PN200-110 show a high degree of stereoselectivity in the inhibition of catecholamine secretion evoked by high K or by DMPP in the presence of Na, the (+)-enantiomer being 57 and 80 times more potent, respectively, than the (-)-enantiomer. Both, noradrenaline and adrenaline release were equally depressed by PN200-110. 3. The IC50 values for (+)- and (-) PN200-110 for blockade of the secretory response induced by K or DMPP in the presence of Na are in the same range. In the absence of Na, (-)-PN200-110 did not affect DMPP-evoked secretion; however, the (+)-enantiomer partially inhibited it. 4. The results suggest that the physiological catecholamine release from chromaffin cells is preceded by Na entry through the nicotinic receptor associated ionophore; this causes cell depolarization, opening of voltage dependent, dihydropyridine-sensitive Ca channels and Ca entry into the cell. In the absence of Na, additional Ca influx through an alternative pathway (the nicotinic cholinoceptor ionophore?) might also activate secretion. PMID- 2975523 TI - Vasoconstrictor and vasodilator effects in normal and atherosclerotic conscious rabbits. AB - 1. Rabbits were fed a cholesterol-rich diet for 5 two-week intervals. Polyvinyl catheters were then implanted into the femoral artery and vein. Dose-response curves to acetylcholine (ACh), noradrenaline (NA), phenylephrine (Phen) and angiotensin II (AII), were obtained in 6 cholesterol-fed and 6 control rabbits before and after isradipine (code name PN200-110) 100 micrograms kg-1. After these experiments the animals were killed and aortic rings were suspended in an organ bath. ACh but not nitroprusside-induced relaxation was impaired in atherosclerotic but not in control preparations. 2. ACh decreased blood pressure dose-dependently in both groups of rabbits even though ACh did not relax the aortae of the same rabbits in vitro. 3. Blood pressure effects reflect mostly changes in resistance vessels. The pressor effects of NA, Phen and AII were enhanced in atherosclerotic compared with normal rabbits. 4. After a dose of 100 micrograms kg-1 isradipine the dose-response curves of all agents were shifted to the right. The differences between atherosclerotic and control rabbits disappeared, except for the AII-induced pressor response, which remained enhanced in atherosclerotic animals. The calcium antagonist thus only partly corrected the atherosclerosis-associated hyperresponsiveness to vasoconstrictor agents. PMID- 2975524 TI - Rat hypothalamic arcuate neuron response in electroacupuncture-induced analgesia. AB - Electroacupuncture (EA) effects on activity of arcuate neurons of the hypothalamus (ARH) and on magnitude of the digastric electromyogram (dEMG) in the jaw opening reflex were investigated, in both p-chlorophenylalanine pretreated and normal Wistar rats. EA stimulation (300-500 microA, 5 msec pulses, for 15 min) was delivered unilaterally to a meridian Ho-Ku point of anesthetized rats at 3, 45 and 100 Hz. In control animals, EA stimulation at 3, 45 or 100 Hz induced long-lasting suppression of the magnitude of the dEMG activity and changed the spontaneous firing rate of most of the ARH neurons: the rate either increased (type I) or decreased (type II). After low-frequency stimulation, there were significantly more type I neurons than type II; after high-frequency stimulation, there were significantly more type II neurons than type I. In serotonin-depleted rats, however, high-frequency stimulation suppressed dEMG activity only slightly and induced a smaller proportion of type II neurons. PMID- 2975526 TI - Kinetic properties of the [Ca2+ + Mg2+]-ATPase in Alzheimer and normal fibroblasts at low free calcium. AB - Kinetic properties of the [Ca2+ + Mg2+]-ATPase from one Alzheimer (AG0364B, 53 year-old donor) and one control fibroblast cell line (AG6009, 59-year-old donor) were examined. A saturation plot revealed the control fibroblast ATPase activity to saturate at approximately 500 nM free Ca2+; whereas, the Alzheimer activity saturated at approximately 1000 nM. Eadie Hofstee graphical analysis indicated almost identical Vmax values, 19.5 and 21.0 nmoles phosphate released/min/mg protein for the Alzheimer and control fibroblasts, respectively. However, an approximately two-fold higher Km value of 350 nM was observed for the Alzheimer fibroblast homogenate in contrast to 220 nM for the control. In a previous study, a kinetic difference in Alzheimer fibroblast [Ca2+ + Mg2+]-ATPase activity at high free Ca2+ concentration was observed. The difference in affinity reported in this study at low free Ca2+ concentrations supports further the hypothesis that abnormalities in Ca2+ homeostasis might be involved in the etiology of Alzheimer's disease. PMID- 2975525 TI - The effects of immobilization stress on beta-endorphin levels are modulated by testosterone. AB - Immunoreactive beta-endorphin (IR-BE) levels were determined in the anterior pituitary (AP), neurointermediate lobe of the pituitary (NIL) and the hypothalamus of castrated male rats and castrated male rats treated with testosterone proprionate (TP), subsequent to exposure to acute (once for 45 min) or chronic (45 min each day for 15 consecutive days) immobilization stress. Acute stress resulted in a reduction in the concentration of IR-BE in the AP of castrated male rats, which was potentiated by TP. The concentration of IR-BE in the NIL was elevated by acute stress in castrated male rats and was not affected by acute stress in castrated male rats administered TP. Exposure to chronic immobilization stress elevated the concentration of IR-BE in the AP of castrated animals and not animals treated with TP. The concentration of IR-BE in the NIL of castrated animals was not altered by chronic immobilization. Chronic stress did result in a significant rise in the level of IR-BE in the NIL of castrated male rats given TP. Hypothlamic IR-BE levels in castrated male rats were reduced by TP and were not influenced by acute or chronic stress. Chromatographic analysis indicated that acute and chronic stress promoted the accumulation of beta lipotropin rather than beta-endorphin in the AP. This effect was attenuated by TP. Beta-endorphin was the only form of immunoreactivity detected in the NIL and hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975527 TI - [General anesthesia in pedodontics]. PMID- 2975528 TI - A comparison of the transabdominal and transvaginal methods of ultrasound-guided aspiration of pre-ovulatory oocytes for in vitro fertilisation. AB - The efficiency of two methods of ultrasound-guided aspiration of pre-ovulatory oocytes was compared in 238 treatment cycles, of which 166 were by the transabdominal-transvesical (TA) route and 72 by the transvaginal (TV) route. Successful collection of at least one oocyte was achieved in 86% of TA attempts and in 97% of TV attempts. Significantly less oocytes were collected from follicles by the TA methods than by the TV methods: 374 oocytes were obtained from 797 follicles (46% recovery) and 242 oocytes from 338 follicles (72% recovery), respectively. Although fertilisation occurred more frequently in TA recovered oocyte (70% versus 54%), the incidence of cleavage per inseminated oocytes collected by these TA and TV methods was similar (49% versus 44%, respectively). Results of this study therefore suggest that the TV method is superior to the TA approach for oocyte recovery. PMID- 2975530 TI - Effects of felodipine on natriuresis, atrial natriuretic factor, the renin angiotensin-aldosterone system, and blood pressure in essential hypertension. AB - The aim of this study was to evaluate the short-term and long-term effects of felodipine, a new dihydropiridine calcium antagonist, on arterial blood pressure (BP), the renin-angiotensin-aldosterone system, diuresis, natriuresis, and the atrial natriuretic factor (ANF). In 15 essential hypertensives (WHO class II) BP, venous BP at the posterior tibial vein (VBPTV), diuresis, natriuresis, plasma renin activity (PRA), and both plasma aldosterone and ANF levels were evaluated at the end of a washout period and after two and 24 hours and 30, 90, and 180 days of follow-up with felodipine, 5 mg twice daily. The first dose of felodipine induced a significant decrease in BP, which was associated with increases in both heart rate and VBPTV. An acute diuretic and natriuretic effect, increases in ANF and PRA, and a transient decrease in plasma aldosterone levels were also observed. Throughout the follow-up period, the antihypertensive efficacy remained unchanged, whereas variations in electrolyte balance and hormonal parameters quickly disappeared, except for the increase in PRA, which lasted until the 30th day of therapy. In our study, felodipine showed a great antihypertensive activity during both short-term and long-term treatment. Moreover, the effect of the first dose was characterized by transient increases in circulating ANF and decreases in plasma aldosterone concentrations, which were associated with marked diuresis and natriuresis. PMID- 2975531 TI - [Atrial natriuretic factor in liver cirrhosis. The effect of volume loading]. PMID- 2975529 TI - Acetyl-L-carnitine acutely administered raises beta-endorphin and cortisol plasma levels in humans. PMID- 2975532 TI - Depression by Fc gamma receptor ligands of SRBC-induced IgM-PFC generation in human blood mononuclear cell cultures. AB - A tissue-culture system to stimulate human peripheral blood mononuclear cells (PBMC) has been employed in which IgM plaque-forming cell (IgM PFC) generation in response to sheep erythrocytes (SRBC) is dependent on macrophages and T suppressor and helper lymphocytes. In this system PBMC from normal subjects give IgM PFC responses ranging from 26 to 938 PFC/culture. Heat-aggregated human IgG or immune complexes present for the duration of culture induce a significant depression of PFC. Unaggregated IgG has no effect on the response or only a moderate stimulatory effect at the highest dose. The results of these experiments are compatible with previous results in a murine system, which indicated that Fc gamma receptor-positive (Fc gamma R+) cells in the suppressor subset are the target for aggregated IgG and induce depression of the PFC response. A similar mechanism may be operating in the human system described here, although the target cell has not been identified. These results may reflect a mechanism of immunomodulation dependent on interaction of Fc receptor (FcR) ligands with FcR, which may play a role in the pathogenesis of immune complex disorders. PMID- 2975533 TI - Acute coronary occlusion after percutaneous transluminal coronary angioplasty. PMID- 2975534 TI - Coronary artery spasm during balloon angioplasty in a patient receiving dopamine infusion. PMID- 2975535 TI - Immunologic effects on peripheral lymphoid cells from patients with chronic hepatitis type B during administration of recombinant interleukin 2. AB - Nine patients with chronic hepatitis type B were treated with recombinant interleukin 2 (rIL-2). Side effects were limited to low-grade fever and headache, which were transient and tolerable for the patients. Seven normal volunteers and nine patients with chronic active hepatitis were administered by one bolus of 500 units of rIL-2. Acute effects of rIL-2 administration on lymphoid cells included a rapid decrease in lymphocytes, especially in cytotoxic T cells and natural killer cells. These acute effects resolved within 24 hours. There was no difference in the changes of immunological parameters between normal volunteers and patients. The same effects were seen during 28 days of rIL-2 administration. The number of lymphocytes and CD4 positive cells was increased after rIL-2 administration for 28 days (P less than 0.01). Natural killer cell activity, especially that of CD16+ and Leu-7- cells was also increased (P less than 0.05). These effects may favour the elimination of virus-infected hepatocytes. PMID- 2975536 TI - Phenotypic expression of Vicia villosa binding T cell subsets, as markers of contrasuppressor cells in systemic lupus erythematosus. AB - The hypothesis that autoimmune diseases might be due to a defect in immunoregulation was tested in systemic lupus erythematosus (SLE). We have applied the double immunofluorescence, flow cytometry technique to peripheral blood lymphocytes from patients with SLE. T cells were studied for their binding of the lectin Vicia villosa (VV) which is a phenotypic marker for contra suppressor cells both in mice and humans. A significant increase in CD3+VV+ and CD8+VV+ cells was found in patients with SLE, as compared with age and sex matched controls (P less than 0.01). When the patients were divided according to the 'lupus activity criteria count', those with active disease had a significantly increased proportion of CD3+VV+ and CD8+VV+ cells, as compared with those showing no disease activity (P less than 0.001). Indeed, a sequential investigation showed that the proportion of CD8+VV+ cells changed in parallel with exacerbation and remission of disease activity. These results suggest that disease activity in SLE is associated with an increase in VV binding CD8 cells which can function as contrasuppressor cells. PMID- 2975537 TI - Associated expression of CD1 antigen and Fc receptor for IgE on epidermal Langerhans cells from patients with atopic dermatitis. AB - The presence of Fc receptors for IgE on epidermal Langerhans cells (LC) from patients with atopic dermatitis (AD) was demonstrated by three different types of experiments. Firstly, cell-bound IgE on LC was removed by acid elution and restored by highly purified human myeloma IgE (IgE kappa). Secondly, after pepsin digestion of cell-bound IgE the number of LC staining with anti-human light chain (kappa, lambda) antibodies significantly decreased in contrast to the number of LC staining with anti-human epsilon heavy chain antibody. Thirdly, LC formed rosettes with sheep red blood cells (SRBC) coated with IgE kappa. Epidermal LC from normal non-atopic controls, did not form rosettes with SRBC-IgE. The SRBC IgE rosette formation could be inhibited by preincubation with IgE kappa and BB10 (MoAb directed against the Fc receptor for IgE on human eosinophils, platelets and macrophages), but also with human IgG, whereas the SRBC-IgG rosette formation could be inhibited neither by IgE kappa nor by BB10. Both the SRBC-IgE and the SRBC-IgG rosette formation could be inhibited by OKT6 (anti-CD1) antibody. The results of inhibition studies with OKT6 antibody on the reconstitution of IgE on epidermal LC after acid elution suggest an associated expression of the CD1 antigen and the Fc receptor for IgE. PMID- 2975538 TI - Modulation of cellular anti-tumour responses by tumour-specific T suppressor lymphocytes. AB - Tumour-specific Ts cells, induced by a protocol which simulates early stages of tumourigenesis in BALB/c mice by the syngeneic plasmacytoma ADJ-PC-5, suppress the generation of specific cytotoxic T lymphocytes (CTL) in a primary in vitro mixed lymphocyte tumour culture (MLTC) of BALB/c spleen cells against ADJ-PC-5. The influence of these Ts cells on the generation of non-specific killer cell activity has now been analysed. The data show that non-specific killer cells generated during in vitro MLTC lyse both ADJ-PC-5 and YAC-1 cells. They are different from tumour-specific CTL as shown by cold target inhibition experiments and on the basis of their different phenotype. The generation of non-specific killer activity during MLTC can be completely suppressed by ADJ-PC-5 specific Ts cells. Activation of non-specific killer cells by recombinant interleukin-2 (r IL2) alone is not influenced by these Ts cells. The in vitro data are best explained by assuming that the target for suppression are ADJ-PC-5 specific T helper cells. Their inactivation will result in depletion of IL2 in the culture, which is required to activate non-specific killer cells. Prevention of activation of specific and non-specific killer cells in vitro by activated tumour-specific Ts cells in the presence of the tumour, suggests that similar mechanisms might operate in vivo and that inactivation of tumour-specific T helper cells might be of central relevance. PMID- 2975539 TI - Selective in-vitro activation of BALB/c T suppressor lymphocytes specific for syngeneic ADJ-PC-5 plasmacytoma cells. AB - T suppressor (Ts) lymphocyte activation is the first reaction of the immune system under conditions which simulate early stages of tumourigenesis. There is evidence that the Ts cell inducing antigen on ADJ-PC-5 plasmacytoma cells of BALB/c origin is a self antigen, which is also expressed on normal BALB/c spleen cells. In order to study this Ts cell inducing determinant in more detail we have developed an in vitro system which allows the selective activation of ADJ-PC-5 specific Ts cells in the absence of activation of cytotoxic T (Tc) cells. The system now allows us to study the modulation of Ts cell induction and function. PMID- 2975541 TI - IL-2 production in the autologous mixed lymphocyte reaction of patients with juvenile rheumatoid arthritis. AB - Interleukin 2 (IL-2) production in the CD4/non-T cell autologous mixed lymphocyte reaction (AMLR) was estimated for patients with juvenile rheumatoid arthritis (JRA). A cellular IL-2 assay (CILA) consisting of co-cultures of indicator cells, CTLL-2, added to CD4/non-T cell AMLR cultures was used. As both CD4 and CTLL-2 cell proliferation are IL-2 dependent, results of the CILA assay were taken as a reflection of the total IL-2 in the co-culture. By this assay most patients with JRA had normal IL-2 production. However, five patients with polyarticular disease had both a low AMLR and low levels of IL-2 production. Three of those tested showed little improvement in the AMLR on addition of recombinant IL-2, suggesting decreased responsiveness to IL-2 as well. PMID- 2975540 TI - Blastogenic responses to Pneumocystis carinii among patients with human immunodeficiency (HIV) infection. AB - Proliferative responses by human peripheral blood mononuclear cells and rat spleen cells were measured to Pneumocystis carinii in a blastogenic assay using organisms obtained from rat lungs and propagated in tissue culture as the antigen. Responses occurred only in subjects with known prior exposure to P. carinii and the magnitude of the response varied with the number of organisms present in the antigen preparation and days in culture. Healthy human adults showed higher proliferative responses to P. carinii than did patients with Class II (asymptomatic) or Class III (lymphadenopathy) human immunodeficiency virus (HIV) infection. No responses to the preparation were found among Class IV HIV patients with the acquired immunodeficiency syndrome, including those with prior episodes of P. carinii pneumonia or those receiving azidothymidine. Overall, the blastogenic responses obtained with P. carinii were similar to those obtained with tetanus toxoid and phytohemagglutinin, and correlated well with the number of circulating CD4 cells. The data suggest that the blastogenic assay using tissue culture-derived rat P. carinii is specific for the organism and should be helpful in studying the cellular immune responses in pneumocystosis among different human patient populations. PMID- 2975542 TI - Analysis of the responding and stimulating cells in the AMLR of patients with rheumatoid arthritis using limiting dilution. AB - Plastic adherent and non-adherent mononuclear cells derived from synovial fluid were found to stimulate peripheral blood mononuclear cells of patients with rheumatoid arthritis (RA) in an autologous mixed lymphocyte reaction (AMLR). Stimulation produced by adherent cells was consistently greater than that produced by non-adherent mononuclear cells. CD11 enriched cells were not significant stimulators in the AMLR, however their presence was required for a significant reaction to occur. Limiting dilution studies revealed that the most plausible model involved two or more cells of the same type in each of the stimulator and responder populations for the AMLR to occur. PMID- 2975543 TI - Communication, the letter, and the GP. PMID- 2975544 TI - Mechanism of action of sulphonylureas with special reference to the extrapancreatic effect: an overview. PMID- 2975545 TI - Diabetic renal disease in central Africa. AB - Six hundred African diabetic patients were examined using a protocol based on the WHO Multinational study in which no country from Africa was represented. The salicylsulphonic acid test for proteinuria was used to assess the presence of diabetic renal disease. Overall 23.8% of patients had proteinuria (95% confidence interval 20.4 to 27.2) and 3.8% chronic renal failure (95% confidence interval 2.3 to 5.3). Patients with proteinuria were older and had had diabetes longer than those without (p less than 0.001). Systolic blood pressure rose with increasing proteinuria in both sexes but only men with severe nephropathy showed an increase in diastolic pressure. Minimal diabetic nephropathy was more common than severe nephropathy which carried a particularly poor prognosis in African diabetic patients due to lack of resources. PMID- 2975546 TI - Blood glucose control and visual and auditory attention in men with insulin dependent diabetes. AB - Assessment of 16 young men with insulin-dependent diabetes mellitus was undertaken with neuropsychological measures of attention, decision-making, and motor tasks. Those patients in very good, or near-normal, blood glucose control demonstrated decreased attention on visual and auditory simple reaction-time tasks, compared with those in moderate blood glucose control. Patient groups did not differ in their decision-making performance; nor did they differ in motor skill performance. These results replicate earlier findings of differences in visual attention in groups of patients according to degree of blood glucose control and show between-group differences with an auditory reaction time measure. The results suggest that patients with near-normal blood glucose control may exhibit slowed simple attention, whether information presentation is visual or aural. PMID- 2975547 TI - Efficacy of dietary regulation in primary health care patients with hyperglycaemia detected by screening. AB - The efficacy of dietary regulation was examined in 38 consecutive primary health care patients with hyperglycaemia detected on screening. Ten weeks of dietary regulation reduced overall mean fasting blood glucose from 8.2 to 6.5 mmol l-1. Fasting blood glucose fell more (from 12.3 to 7.6 mmol l-1 and from 8.4 to 6.6 mmol l-1) in the two quartiles initially above the median (7.15 mmol l-1), than in the lower quartiles (6.7 to 6.1 mmol l-1 and 5.7 to 5.8 mmol l-1), even though weight reduction was similar. The reduction in blood glucose correlated (r = 0.87) with the degree of fasting hyperglycaemia before treatment. Sixteen patients (42%) reached or maintained fasting blood glucose less than or equal to 6.0 mmol l-1, and they had a milder degree of glucose intolerance, a higher insulin response to a meal and a greater reduction in weight than the 22 patients (58%) who did not reach less than or equal to 6.0 mmol l-1. Nine patients (24%) maintained fasting blood glucose less than or equal to 6.0 mmol l-1 for greater than 5 years, and showed a considerable improvement of insulin action. Dietary regulation improved glucose control mainly by reducing fasting hyperglycaemia; neither the delay in early insulin release nor the associated elevation and prolongation of the post-prandial glucose excursions were reduced. PMID- 2975548 TI - Screening for diabetes: does measurement of serum fructosamine help? AB - The diagnostic sensitivity and specificity for diabetes of serum fructosamine levels and fasting venous blood glucose concentrations were compared in 613 subjects during a diabetes community screening programme of 1049 adult Muslim Asians in Dar es Salaam, Tanzania. Using WHO (1985) criteria 228 had impaired glucose tolerance (IGT), 41 had previously been diagnosed as having diabetes while 32 had newly recognized diabetes. The mean (+/- SD) serum fructosamine levels were 20.9 +/- 3.2, 21.6 +/- 3.2, 23.9 +/- 4.9, and 30.1 +/- 7.9 (mumol g-1 albumin) in subjects with normal glucose tolerance, IGT, newly diagnosed diabetes, and previously diagnosed diabetes, respectively (p less than 0.001 for differences between groups). The specificity of values above the mean +2SD normal was 99% for abnormal glucose tolerance with a sensitivity of only 22% for diabetes. The predictive values were 44% and 97% for positive and negative results, respectively. Very little difference from normal was found for IGT subjects. Expressing fructosamine values in absolute terms or per gram albumin made little difference to sensitivity and specificity. The sensitivity was only 32% for fasting blood glucose greater than or equal to 6.7 mmol l-1, 73% for values greater than or equal to 5.5 mmol l-1, and 100% for fasting blood glucose greater than or equal to 4.5 mmol l-1. It is concluded that both serum fructosamine and fasting blood glucose are poor screening and diagnostic tests for diabetes and for IGT, and that glucose loading is required. PMID- 2975549 TI - The effect of glibenclamide and metformin on serum lipoproteins in type 2 diabetes. AB - In a cross-over study, the effects of 3 months treatment with metformin or glibenclamide on body weight, blood glucose control, and serum lipoproteins were compared in 35 Type 2 diabetic patients, inadequately controlled by dietary therapy alone. Glibenclamide alone increased body weight (mean change +2.75 kg; 95% confidence intervals +1.95 to +3.55 kg; p less than 0.0001). Glibenclamide and metformin achieved equivalent blood glucose control, independent of initial body mass index. Neither drug affected serum triglyceride concentration. Metformin alone significantly reduced low density lipoprotein cholesterol (mean change -0.34 mmol l-1; 95% confidence intervals -0.12 to -0.57 mmol l-1; p less than 0.01). Neither drug altered high density lipoprotein or subfraction cholesterol. PMID- 2975550 TI - Recurrent diabetic ketoacidosis. AB - In a study over 15 years, 740 episodes of diabetic ketoacidosis occurred in 505 patients. A total of 113 patients had more than one episode. The majority (90%) of recurrences were within 4 years of the initial episode. Taking a definition of recurrent ketoacidosis as three or more episodes within 4 years, 39 patients were identified. Two subgroups appeared prone to such frequent recurrences, namely girls less than 20 years of age and women more than 59 years of age. A variety of causes of recurrent episodes was noted in the young patients but in the older patients other chronic illnesses complicated the diabetes. The need for good domiciliary supervision of elderly chronic sick patients who require insulin is emphasized. PMID- 2975551 TI - The variability in the action of unmodified insulin is more dependent on changes in tissue insulin sensitivity than on insulin absorption. AB - Eight normal subjects were studied twice for 360 min after the subcutaneous injection of unmodified insulin (0.15 U kg-1) during euglycaemic clamps. Insulin absorption was assessed by both the area under the insulin-time curve above baseline (AUC) and time course of absorption (time to 25% and 50% of total AUC). Insulin action was measured as the amount of glucose infused. The maximal serum insulin concentration was 0.27 +/- 0.02 (+/- SE) nmol l-1 at 112 +/- 10 min. Fifty percent of total glucose infused occurred at 218 +/- 7 min. The maximal glucose infusion rate was 5.11 +/- 0.70 mg kg-1 min-1 and occurred at 256 +/- 12 min. Intrasubject coefficients of variation (CV) for total insulin AUC (11.2%), time to 25% of maximum AUC (12.1%) and time to 50% of maximum AUC (10.2%) were considerably lower than that for total insulin action (22.6%). Total insulin AUC did not correlate with total glucose utilization (r = 0.06, NS). We conclude that when glucose concentrations are maintained by euglycaemic clamps the peak of unmodified insulin action is later and the duration longer than traditionally recognized, insulin AUC does not predict insulin action, and the higher variability of insulin action compared with the indices of absorption suggests that day-to-day changes in tissue insulin sensitivity contribute more to the variability in insulin action than changes in absorption. PMID- 2975552 TI - Anti-smoking advice for young diabetic smokers: is it a waste of breath? AB - The effect of health counselling on the smoking habits of 60 diabetic patients (aged less than 40 years) was assessed. Measurement of breath carbon monoxide (CO) and urinary cotinine, a metabolite of nicotine, were used as objective markers of smoking. All patients wished to cease smoking and the impact of health counselling was reviewed in a 'Stop Smoking' clinic. In addition to routine advice on the health hazards of smoking, half the patients and their families also received further counselling during a home visit by a health visitor. After 6 months many of the 60 patients claimed to have reduced their cigarette consumption. However, the urinary cotinine concentrations did not confirm this. Only one patient actually stopped smoking and he had sustained a myocardial infarction during the study. There was a small but significant reduction of breath CO in the patients seen at home by the health visitor but the urinary cotinine concentrations were unchanged. This suggests that these patients abstained from smoking for only a few hours before attending the 'Stop Smoking' clinic. PMID- 2975553 TI - Improvement in morning hyperglycaemia with basal human ultratard and prandial human actrapid insulin--a comparison of multiple injection regimens. AB - Three 'pen'-administered multiple injection regimens have been compared with twice daily insulin injection regimens by means of 24-h profiles of plasma glucose and free insulin concentrations. Ten Type 1 diabetic patients received their usual twice daily insulin regimen and were then randomized to receive the same total daily insulin dose in four divided doses using (1) 50:50 premixed soluble and isophane, (2) 30:70 premixed soluble and isophane, and (3) preprandial soluble and evening crystalline-zinc insulins. Profiles were performed after 1 week on each regimen. Plasma glucose concentrations were similar during the twice daily regimen and the two premixed regimens, rising during the early hours of the morning to a peak between 0900 and 0930 h of 13.8 +/- 2.8 (+/- SD) mmol l-1 on the twice daily regimen, 13.6 +/- 5.3 mmol l-1 on the premixed 50:50 regimen, and 13.5 +/- 4.2 mmol-1 on the premixed 30:70 regimen. With the basal and prandial regimen, overnight plasma glucose concentrations were higher than with the other regimens between 2400 and 0300 h (p less than 0.05). Concentrations then fell until breakfast, and rose after this meal to a peak of 9.5 +/- 4.3 mmol l-1 (p less than 0.01). Mean plasma glucose concentrations were significantly lower than on the other three regimens between 0830 and 1100 h (p less than 0.05). Less variability was observed in 24-h mean plasma glucose concentrations during the basal and prandial regimen than during the other three regimens. PMID- 2975555 TI - Giving dietary advice to Asian diabetic patients. PMID- 2975554 TI - Sucrose versus saccharin as an added sweetener in non-insulin-dependent diabetes: short- and medium-term metabolic effects. AB - Seventeen non-insulin-dependent diabetic patients were randomly allocated to their usual diet supplemented daily with either 28 g sucrose or 30 g starch (isoenergetic with sucrose) and saccharin (equivalent sweetness). After 6 weeks, the supplements were reversed. No significant treatment effects were observed on fasting concentrations of blood glucose, plasma insulin or serum triglycerides, or on urinary excretion of glucose, sodium or potassium. Following a standard breakfast with either sucrose or saccharin and starch, no differences between meal responses were observed. This study demonstrates no medium-term metabolic contraindications to including a moderate amount of sucrose in the diets of patients with non-insulin-dependent diabetes mellitus. PMID- 2975556 TI - A team approach to developing a district diabetic service. PMID- 2975557 TI - The Northern Ireland Diabetes Group. PMID- 2975558 TI - Uncontrolled hypertension in type 1 diabetes: assessment of patients' desires about treatment and improvement of blood pressure control by a structured treatment and teaching programme. AB - Control of arterial blood pressure at near-normal levels is of importance for the prognosis of patients with Type 1 (insulin-dependent) diabetes mellitus. In non diabetic populations patient compliance to antihypertensive therapy is frequently poor, especially in young people. Thirty-seven consecutive eligible patients with longstanding Type 1 diabetes and persistently uncontrolled hypertension were questioned about their preferences regarding hypertension treatment. Throughout they expressed a strong desire for more information about hypertension and for active participation in monitoring of blood pressure and therapeutic decision making. In addition, they showed considerable reluctance to accept a pharmacological intervention. Therefore, in order to improve blood pressure control, the patients participated in a structured outpatient hypertension treatment and teaching programme for groups of about 6 patients consisting of four teaching sessions at weekly intervals. The programme comprised home monitoring of blood pressure and involvement of the patients in treatment decision-making. In 34 patients who had a complete follow-up examination after an average of 16 months, mean sitting arterial pressure had decreased from 111 to 101 mmHg (p less than 0.001) and 53% of the patients were below 140/90 mmHg. The number and dosage of prescribed antihypertensive agents remained unchanged by the intervention. Sodium intake was not reduced during the study, but pulse rate decreased significantly in patients treated with beta-adrenergic blockers and serum uric acid rose in patients on diuretic therapy, suggesting increased adherence of the patients to prescribed antihypertensive drug therapy. In 34 comparable Type 1 diabetic patients who were not subjected to a hypertension treatment and teaching programme mean arterial blood pressure remained unchanged during a 12-month period. PMID- 2975559 TI - Questionnaire design in diabetes care and research. 1: Getting the question right. PMID- 2975560 TI - Extra-laboratory blood glucose measurement: a policy statement. AB - The introduction of test strips for measurement of blood glucose has led to their widespread use by medical and paramedical staff outside the central laboratory. While many users have developed ad hoc schemes of training and quality control, formal guidelines have not been presented. This paper makes recommendations on various aspects of extra-laboratory blood glucose measurement in hospitals, but not in the home, with the aim of improving the safety and reliability of results. It is recommended that: 1. a formal programme of training should be devised by staff in the laboratory together with those in the diabetes service and others as appropriate; 2. laboratory and diabetes staff should train, assess, and approve more senior users who would then take responsibility for the training and performance of more junior colleagues; 3. all users should demonstrate their competence at regular intervals; 4. there should be a quality assurance programme arranged by the laboratory which tests the performance of the operator and the equipment; 5. laboratory staff, in consultation with users, should be responsible for the selection and purchase of equipment; 6. equipment should be cleaned and maintained on a regular basis by users. PMID- 2975561 TI - Psychological factors and metabolic control: time for collaboration. PMID- 2975562 TI - Development of implantable insulin pumps: long is the road. PMID- 2975563 TI - Diabetes centres: an important development or another gimmick? PMID- 2975564 TI - Immunotherapy in type 1 diabetes. PMID- 2975565 TI - Measuring psychosocial adaptation to diabetes in adolescence. AB - We have developed a psychometric test to assess psychosocial adaptation to diabetes in young people aged from 10 to 17 years. This includes subscales which describe emotional difficulty with, and attitude to, diabetes. The content of the scales was drawn from unstructured discussions with 27 young people with diabetes. The scales were piloted on two further unselected groups of adolescents (n = 50, n = 99) to examine their reliability, validity, and acceptability. Highly significant correlations were found between the results of the scales and measures of psychosocial function (p less than 0.001), parents' perceptions. Performance on the scale also correlated with a number of behaviour variables and differentiated those who were reported to be restricted by their diabetes (p less than 0.005) and those who had had a previous psychiatric referral (p less than 0.05). The scales appear reliable within the constraints of the small population examined. PMID- 2975566 TI - Reduced free radical activity detected by decreased diene conjugates in insulin dependent diabetic patients. AB - Free radicals are unstable chemical species which react with and oxidize adjacent molecules, particularly polyunsaturated lipids. The diene-conjugated non-peroxide isomer of linoleic acid (PL-9,11-LA') has been identified as the main diene conjugated compound in plasma, and is a probable marker of free radical activity. The aim of the current study was to determine whether the level of PL-9,11-LA', measured by HPLC, is altered in insulin-dependent diabetes, and to investigate whether any abnormality demonstrated correlated with microvascular disease in the form of retinopathy. There was no difference in the concentrations of linoleic acid between the diabetic and control groups (422(129) vs 402(81) (SD) mumol l 1). However, the concentration of PL-9,11-LA' was significantly reduced in the diabetic group compared with control group (15.6(6.7) vs 19.3(3.9) mumol l-1, p less than 0.01), with the molar ratio of PL-9,11-LA':linoleic acid x 100 similarly reduced (3.8(1.3) vs 5.0(1.6)%, p less than 0.005). This study does not support the concept that free radicals play a significant role in the development of diabetic vascular disease. PMID- 2975567 TI - A randomized trial of the efficacy and acceptability of a pen injector. AB - A controlled trial of pen injection of insulin was performed in 78 patients, with assessment of metabolic control and lifestyle. After a 6-week run-in period, during which control was optimized, the patients were randomized, either to stay on a twice daily insulin regimen (n = 37), or to change to a three times daily pen regimen with human ultralente at night (n = 41). Over the 20 weeks, there was no significant change in mean glycosylated haemoglobin (syringe, mean +/- SD, 11.1 +/- 2.5% to 10.9 +/- 2.0%; pen, 11.3 +/- 2.6% to 11.2 +/- 2.0%), in blood glucose profiles or in frequency of hypoglycaemic attacks in either group. A self completed questionnaire demonstrated high patient satisfaction with the pen injector (NovoPen), 78% for effect on lifestyle and 81% for increased flexibility. Ninety-five percent preferred the pen injector regimen to conventional treatment and stayed on it. PMID- 2975568 TI - A mixed meal potentiates the insulin sensitivity of glucose transport and metabolism in adipocytes from patients with type 2 diabetes mellitus. AB - Post-glucose enhancement of insulin action may represent a physiological mechanism for the acute regulation of insulin sensitivity of target tissues. To clarify whether a similar mechanism is operative in the insulin-resistant diabetic state we have investigated the effects of a mixed meal on adipocyte insulin action in eight patients with Type 2 diabetes mellitus. Ninety minutes after ingestion of breakfast insulin binding to fat cells increased by 21% (p less than 0.05). In the fasting state 6 patients had a significant response of glucose transport and lipogenesis to insulin whereas two exhibited non responsiveness. In the 6 responders insulin sensitivity, as estimated by the insulin concentration at which half-maximal effect was achieved, increased for glucose transport (before, 260 +/- 46 pmoll-1; after, 105 +/- 21 pmol l-1; p less than 0.05) and for lipogenesis (before, 36 +/- 9 pmol l-1; after, 9 +/- 2 pmol l 1; p less than 0.05). No significant changes occurred in basal or maximal glucose transport or lipogenesis. In the two primary non-responders intake of the meal was associated with average increase in maximal insulin responsiveness of 52% for glucose transport and 28% for lipogenesis. Intake of a mixed meal is associated with a slight increase of insulin binding to adipocytes from patients with Type 2 diabetes mellitus but a marked increase of adipocyte insulin sensitivity at the post-binding levels of glucose transport and metabolism. PMID- 2975569 TI - Predicting future treatment of diabetes mellitus from characteristics available at presentation. AB - This study was designed to identify the clinical features of a newly diagnosed diabetic patient that are most useful in deciding treatment. A secondary aim was to formulate a statistical model for predicting subsequent treatment. The following features were considered in 289 patients: age, sex, severity and duration of symptoms, degree and duration of weight loss, glycosuria, ketonuria, blood glucose concentration, body mass index (BMI), and family history of diabetes. Three treatment groups, 6 months after diagnosis, were defined: diet alone, diet with oral hypoglycaemic agent(s), and insulin-treated. Univariate analysis showed that symptom severity, glycosuria, ketonuria, glucose concentration, weight loss, and BMI were significantly different between the three groups. Age and rate of weight loss were significantly different between the insulin-treated and non-insulin-treated groups. Multivariate analysis gave a model to calculate the probability of requiring each of the three treatments given certain characteristics. A second cohort of 174 patients was used to assess the accuracy of the model. The model predicted the actual treatment at 6 months correctly in 72%. PMID- 2975571 TI - Insulin antibodies do not influence the absorption rate of subcutaneously injected insulin. AB - The influence of insulin antibodies on absorption rate and plasma free insulin concentrations after subcutaneous injection of insulin, was studied in two groups of insulin-treated diabetic patients, one without insulin antibodies (n = 9) and a second with high plasma concentrations of antibodies (n = 14). Except for antibody concentration there were no differences in clinical variables. During 8 h after the injection of 12 U of iodinated neutral human insulin, residual radioactivity at the injection site, plasma glucose, and free and total insulin were measured. Significant differences in absorption rate of insulin were not found between the groups. Plasma glucose (basal value 16.8 +/- 4.4 SD vs 16.1 +/- 4.2 mmol l-1) and free insulin (basal value 8.3 +/- 1.4 vs 11.4 +/- 2.3 mU l-1, maximum after 90 min 36.9 +/- 19.5 vs 30.5 +/- 18.7 mU l-1) were never significantly different between the groups, nor were areas under the curve for free insulin (191.4 +/- 69.2 vs 170.8 +/- 98.6 mU l-1 h). In the high antibody group a small increase in bound insulin was found. PMID- 2975570 TI - Additional factors associated with plantar ulcers in diabetic neuropathy. AB - Patients can only examine and handle their own feet if they have adequate visual acuity and joint mobility. We therefore studied the physical capacity of patients with neuropathy to perform the preventive footcare measures previously taught. The study included three groups of diabetic outpatients, comparable for age and duration of diabetes: (1) 38 patients with neuropathic ulcers; (2) 21 patients with neuropathy, but no ulcers; (3) 30 patients without neuropathy. Visual acuity and joint mobility, expressed as minimum eye-metatarsum and heel-buttock distances, did not differ between uncomplicated neuropathic and non-neuropathic patients: visual acuity was sufficient in 95% of neuropathic patients without ulceration and in 87% of non-neuropathic patients; joint mobility was in the normal range in both groups. However, 71% of complicated neuropathic patients had insufficient visual acuity for correct foot examination, and their joint mobility was reduced compared with uncomplicated neuropathic and non-neuropathic patients. PMID- 2975572 TI - Mechanisms of autoimmunity. PMID- 2975573 TI - Contamination of insulin by silicone oil: a potential hazard of plastic insulin syringes. AB - A 32-year-old insulin-dependent diabetic patient reported recurrent clouding of her short-acting insulin, caused by silicone oil contamination from re-used disposable syringes. PMID- 2975574 TI - Laboratory, nurse and patient assessment of Glucostix blood glucose reagent strips. AB - The new Glucostix (Ames) blood glucose reagent strips have been assessed for accuracy and acceptability by nurses using them on the ward and by patients at home. When used with a Glucometer II, Glucostix showed close correlation with blood glucose measured by Yellow Springs Analyser (r = 0.97, p less than 0.001). Using the strips without a meter patients achieved a good correlation with the Yellow Springs Analyser when measuring their own capillary blood glucose (r = 0.94, p less than 0.001). Ward nurses were less precise (r = 0.87, p less than 0.001) and this, perhaps, reinforces the importance of regular quality control to maintain the precision of blood glucose monitoring using reagent strips. Comparing Glucostix with BM Glycemie 1-44 strips, 14 of 15 nurses preferred Glucostix whereas patients showed no overall preference, 37% preferring BM Glycemie 1-44, 33% Glucostix, and 30% stating no preference. There were only minor differences between the two reagent strips. Both groups considered the Glucostix colour change to be easier to read, particularly when using the perforated bench readers, but some thought Glucostix needed more blood and that it was more difficult to remove from the pad. Thus, the Glucostix system provides an accurate measure of blood glucose and is an acceptable alternative to BM Glycemie 1-44 strips for use at home or on the ward. PMID- 2975575 TI - Compact: a computer package for clinical trials. PMID- 2975577 TI - Do we need IGI? PMID- 2975576 TI - The diabetic foot 1988. PMID- 2975578 TI - Occupational dermatoses in workers exposed to resins based on phenol and formaldehyde. AB - During December 1985 and February 1986, an investigation was carried out into skin diseases among workers in a plant producing decorative equipment built of paper sheets impregnated with resol resins based on phenol and formaldehyde (P-F R). A questionnaire was sent to all 238 employees and it was answered by 218 (91.6%). Previous and current dermatoses were reported by 98 workers and 89 of these were examined and patch tested. The patch testing was conducted with a standard test series and products from the working environment and revealed contact allergy to P-F-R in 9 persons and to formaldehyde in 1. Besides these 10 individuals with occupational allergic contact dermatitis, occupational dermatoses were diagnosed in an additional 20 workers; irritant contact dermatitis in 19 and chemical burn in 1. In total, the figure for occupational dermatoses was 30 (12.6%). PMID- 2975579 TI - Occupational sensitization to epichlorohydrin and epoxy resin. AB - Sensitization to epichlorohydrin (ECH) is reported in 6 patients with occupational contact allergy. In 2 cases, an isolated positive test to ECH was found. In the remaining 4 cases, concomitant positive reactions were seen to "ICDRG epoxy resin" (MW 385) and to liquid epoxy resin (MW 370). Allergy to bisphenol A was not seen in the patients. 5 patients worked in an epoxy resin plant. Adequate preventive measures to avoid skin contact with ECH are required to prevent ECH sensitization becoming a more serious industrial hazard. PMID- 2975580 TI - Occupational dermatitis due to sodium hypophosphite. PMID- 2975581 TI - Degradation of rat chondrosarcoma proteoglycans by a neutral metalloprotease from rabbit chondrocytes. AB - Rat chondrosarcoma proteoglycan aggregate with radiolabeled core protein was digested with a chondrocyte metalloprotease (CMP) or clostripain (CP) at neutral pH. The rates of product formation and the sizes and antigenicities of the products were studied using column chromatography and monoclonal antibodies. Sixteen percent of [35S]methionine label and 17-18% of [3H]serine label in core protein were freed from glycosaminoglycan bound peptides by 50 U/ml (760 micrograms/ml) of CP or 10 micrograms/ml (estimated) of CMP in 180 min. The CP reaction was almost complete at 5 minutes while the CMP reaction proceeded slowly from 5 to 180 min. The chondroitin-sulfate rich fragments were smaller after CP than CMP treatment. The 180 min CMP digest contained protein that migrated in 2 peaks on Sepharose CL6B. These two peaks corresponded to the peaks where hyaluronic acid binding region produced by CP and link protein migrate. Metalloenzyme inhibitors inhibited CMP with IC50s of 5 x 10(-5)M, 1 x 10(-3)M, and 80 micrograms/ml for phenanthroline, EDTA, and alpha 2-macroglobulin, respectively. PMID- 2975582 TI - Effect of two oral contraceptives containing 30 micrograms ethinylestradiol and 75 micrograms gestodene or 150 micrograms desogestrel upon various hormonal parameters. AB - The effect of two oral contraceptives containing 30 micrograms ethinylestradiol + 75 micrograms gestodene or 30 micrograms ethinylestradiol + 150 micrograms desogestrel upon various hormonal parameters were measured in 11 women each on days 1, 10, and 21 of the first, second, third, sixth, and twelfth treatment cycle and compared to the levels on days 1, 10, and 21 of the preceding control cycle. There was no significant difference in the clinical effects or in the influence on the serum hormone parameters between both formulations. A significant decrease in the serum concentrations of luteinizing hormone and follicle stimulating hormone was observed during each cycle which was dependent on the duration of intake. Contrary to this, prolactin was not significantly altered, but 6 out of the 22 women showed episodically elevated prolactin levels. Serum estradiol and progesterone were profoundly suppressed, except one woman who ovulated during the twelfth cycle probably due to a therapy with metamizol, trimethoprim and sulfamethoxazole. The concentrations of dehydroepiandrosterone sulphate were significantly and time-dependently reduced by 20 to 25% during each treatment cycle. There was also a significant decrease in the serum levels of testosterone by 20 to 30% and of free testosterone by 40 to 60%, while sex hormone-binding globulin increased by 250 to 300%. It could be observed that during the pill-free interval of 7 days the pituitary and ovarian function recovered, while the sex hormone-binding globulin levels remained elevated by 100%. PMID- 2975584 TI - Utilisation of health services over a one-year period by an adolescent population with Down syndrome. AB - The utilisation of health services during a one-year period by an adolescent population with Down Syndrome (DS) was surveyed and compared to controls of the same age-group and same geographical area. The probands comprised 28 males (65%) and 15 females (35%). The controls comprised 22 males (40%) and 33 females (60%). Mean age: 17.6 years for probands and 17.4 years for controls. Day of census, 31 December 1985. Chronic diseases or conditions were found to be significantly more frequent among probands than among controls. Controls visited their general practitioner significantly more frequently than the probands, who utilised the services of practising specialists significantly more frequently than controls. DS individuals received significantly more dental care than did controls. Altogether, probands utilise the secondary health services more frequently than controls. The question of sufficient secondary health services for DS-adolescents is discussed, but no conclusion can be drawn from the results of the present study. PMID- 2975583 TI - Acceptability of injectable contraceptives in Assiut, Egypt. AB - The present work was a randomized comparative study of two injectable progestogen only contraceptives. The first group (200 subjects) received 150 mg of depotmedroxyprogesterone acetate (Depoprovera) every 84 +/- 7 days and the second (200 subjects) received 200 mg of norethisterone enanthate (Noristerat) every 56 +/- 7 days. Acceptors of injectable contraceptives in Assiut, Egypt, were mainly women looking for fertility termination. Menstrual disruption was the main side effect among both treatment groups. Amenorrhoea was the commonest menstrual complaint and was the main reason for discontinuation in both groups. Only one pregnancy occurred during NET-EN use; two more pregnancies occurred, one in each of the two groups but there were indications that conception preceded the first injection. Menstrual irregularities were generally more frequent with DMPA users. However, DMPA had better one-year continuation rates than NET-EN (68.8 +/- 3.5 and 57.1 +/- 3.6 per 100 women, respectively). PMID- 2975585 TI - Ischemic papillopathy and contraceptives. PMID- 2975586 TI - Sternocostal joints, low back pain and lumbar discopathy. AB - New interrelations between the distensions of the 5th, 6th and 7th ribs and low back pain were studied in two groups of patients: 1) one hundred out-patients with prevalent pseudoradicular symptomatology and 2) fifty patients hospitalised for graphically verified (CT, PMG) lumbar disc herniation and pronounced radicular symptomatology. We have introduced some measurements to document asymmetrical holding of the body and limbs: the distance between the sternoclavicular joint-spina iliaca ant. sup. and the distance between the acromion and spina iliaca post. sup. We found that in the distension (blockade) of the 5th rib, constant spasms are present in m. abdominis obliquus ext., in the most lateral part of the adductors and in the most medial part of the great gluteal muscle. In 6th rib distension, spasms in the external part of the m. rectus abdominis, and in the mediolateral fascicles of thigh adductors and in the external part of m. glutaeus max. could be palpated. In 7th sternocostal distension there are spasms in the most medial parts of m. rectus abdominis, and of thigh adductors and m. glutaeus medius of the same side. After mobilization, statistically significant improvement of body asymmetry (p less than 0.01), improvement of anteflection, bending to the side, Lasegue's and Patric's phenomena were found. Surgery was indispensable only in 6% of our patients suffering from severe lumbar disc hernianion (in 3 out of 50). The CT appearance of the prolapse remained unaltered even after clinical remission. There were some changes before and after mobilization in the CT picture of sternocostal joints. The importance of strengthening of the great pectoral muscles--in which preexistent relative weakness was found--is stressed. The beneficial effect of mobilization of the respective sternocostal blockade on the pain syndrome, body asymmetry and mobility is described. PMID- 2975587 TI - Dealing with thickened mycotic toenails. PMID- 2975589 TI - [Emergency coronary bypass surgery after failure of percutaneous transluminal coronary angioplasty in 2 cases]. PMID- 2975588 TI - [Alpha-hANP-like immunoreactivity in lung cancer]. AB - Specific radioimmunoassay and immunohistochemical method of alpha-human atrial natriuretic polypeptide (alpha-hANP) was established to diagnose one case of large cell lung carcinoma (clear cell carcinoma) proved by pathology. It was found that there was alpha-hANP-like immunoreactivity in the cancer cells at a concentration of 147.5 pg/mg protein. Chromatography analysis showed that alpha hANP-like immunoreactive materials had multiple molecular forms. The significance of these phenomena remains to be further investigated. PMID- 2975590 TI - [Complicated percutaneous transluminal coronary angioplasty in 2 cases]. PMID- 2975591 TI - [Use of a balloon catheter in percutaneous transluminal coronary angioplasty]. PMID- 2975592 TI - [Atrial natriuretic factor: its physiologic characteristics]. PMID- 2975593 TI - [Streptozotocin-induced non-insulin dependent diabetes mellitus in rats]. PMID- 2975594 TI - Antitumor actions of toremifene in the 7,12-dimethylbenzanthracene (DMBA)-induced rat mammary tumor model. AB - Toremifene (200 and 800 micrograms/day) or tamoxifen (200 micrograms) were effective in preventing the development of 7,12-dimethylbenzanthracene-induced rat mammary tumors when given p.o. from day 28 after carcinogen administration. This antitumor action was completely reversed if the toremifene or tamoxifen treatment was stopped or partially reversed by coadministration of progesterone (4 mg/day). Large doses of toremifene (4000 and 8000 micrograms/day) for 10 days produced very high circulating levels of the parent compound (282 +/- 49 and 1002 +/- 224 ng/ml, respectively) and N-desmethyltoremifene (2631 +/- 449 and 6999 +/- 1308 ng/ml, respectively). However, even these very high levels did not reduce the number of animals ultimately developing tumors or the number of tumors each animal developed following cessation of toremifene administration. These findings indicate toremifene has a tumoristatic rather than tumoricidal action in this tumor model. PMID- 2975596 TI - Back-stimulation of B lymphocytes binding to helper T cell surface antigens. AB - Helper T cells directed to B cell surface determinants activate their "targets" into polyclonal antibody production in vitro and in vivo, but B cells which bind to epitopes on the helper cell surface are preferentially induced. Furthermore, "anti-helper" B cell activation also occurs by "back-stimulation", that is even when the responding B lymphocytes are not specific targets for the inducing helper cells, as long as these are simultaneously activated by appropriate interactions with presenting cells. In our experimental systems, this is the only condition where "bystander" activation can be recorded. These findings suggest mechanisms of auto-antibody production associated with unrelated helper cell activity in vivo. PMID- 2975595 TI - Doxazosin in patients with hypertension. AB - The antihypertensive effects and steady-state pharmacokinetics of doxazosin, as well as the bioequivalence of four dosage forms, were studied in 25 hypertensive patients. For an 8 mg daily dose mean Cmax at steady-state for all patients was 108 ng/ml; the mean tmax was 1.8 h. The mean terminal elimination half-life was 22 h. The four tablets containing 1, 2, 4, or 8 mg of doxazosin were bioequivalent in delivering the 8 mg dose. In patients with mild to moderate hypertension, 26-day treatment with doxazosin resulted in blood pressure reduction of 10/7 mmHg in the supine and 13/18 mmHg in the standing position. Adverse effects were generally mild and of brief duration. PMID- 2975597 TI - Tolerization during pregnancy: impact on the development of antigen-specific help and suppression. AB - Analysis of splenic B cells in the offspring of BALB/c mice which were treated with 2,4,6-trinitrobenzenesulfonic acid (TNBS) throughout the gestational period revealed that reduced responsiveness was predominantly reflecting changes in regulatory cells. Since development of T cells takes place in the thymus, postnatal maturation of helper (Th) and suppressor (Ts) T cells was evaluated in the thymus as well as in the spleen. To elaborate the impact of prenatal TNBS treatment on the trinitrophenyl (TNP) repertoire, TNP-sheep red blood cell- and fluorescein isothiocyanate-specific regulatory elements were determined in parallel. It was found that the thymus contained activated helper and suppressor cells during the first 2 weeks of postnatal life, the majority of which appeared to be self-specific. Self reactivity in thymus and periphery disappeared within 3 weeks. Prenatally TNBS-treated mice differed inasmuch as they apparently recognized TNP-self as a self component, TNP-self reactivity being down-regulated similar to self reactivity in prenatally untreated mice. This explains why prenatally TNBS-treated mice responded poorly to stimulation with conventional T dependent TNP-carrier conjugates. Yet, they responded transiently to TNP-anti-TNP conjugates. It could be shown that this was due to down-regulation of putatively idiotype-specific Ts, which were highly increased in prenatally TNBS-treated mice. Data are interpreted in the sense that the establishment of self tolerance is preceded by activation of self-reactive Th, which become partially eliminated. Remaining autoreactivity is controlled by the establishment on an equilibrium between idiotypic and anti-idiotypic elements. PMID- 2975598 TI - Structural and serological heterogeneity of gamma/delta T cell antigen receptor expression in thymus and peripheral blood. AB - Monoclonal antibodies (mAb) reactive against the gamma/delta T cell antigen receptor (TcR) have been used to characterize the distribution and structural properties of gamma/delta TcR-bearing lymphocytes in blood and thymus. Consistent with prior reports the TcR gamma/delta-1 and delta-1 mAb react with all gamma/delta TcR+ T lymphocytes in blood and thymus. By contrast the TCS-delta mAb was found only to react with a subset of the gamma/delta TcR-bearing T cell population. Several lines of evidence suggest that this reagent preferentially reacts with the V delta 1 gene product. Using these reagents, it was observed that gamma/delta TcR+ T lymphocytes comprise 4.6 +/- 3.5% (range 1.0-16.3%) of peripheral blood lymphocytes. However, analysis of peripheral blood from normal adult donors revealed that in 29 of 32 the TCS-delta (possibly V delta 1)-bearing cells comprised less than 30% of the total gamma/delta-TcR+ population. Biochemical analysis demonstrated that the predominant form of the gamma/delta TcR in adult peripheral blood is a disulfide-linked heterodimer, indicating preferential use of the C gamma 1 gene. The delta TcR chain from these TcR gamma/delta-1+/TCS-delta- T cells was remarkably basic in charge, as analyzed by nonequilibrium pH gradient electrophoresis. By contrast with peripheral blood the majority of freshly isolated and interleukin 2-cultured gamma/delta TcR+ thymocytes were predominantly TcR-gamma/delta-1+/TCS-delta +, and preferentially expressed V delta 1. Moreover, both disulfide-bonded and nondisulfide-bonded gamma/delta TcR heterodimers were expressed in all thymuses examined and both forms were contained within the TCS-delta + thymic subset. Similar to recent findings in the mouse, these studies suggest a possible bias in the structural form of gamma/delta TcR based on tissue location. PMID- 2975599 TI - C1q acts synergistically with phorbol dibutyrate to activate CR1-mediated phagocytosis by human mononuclear phagocytes. AB - The adherence of human monocytes and culture-derived macrophages to surfaces coated with complement subcomponent C1q has been previously shown to enhance Fc receptor (FcR)-mediated phagocytosis by these cells. We examined the effects of C1q on C3b/C4b receptor (CR1)-mediated phagocytosis by mononuclear phagocytes. A small percentage of human monocytes cultured in the presence of serum became competent to ingest sheep erythrocytes bearing IgM and C4b (EAC4b). This phagocytic activity was enhanced when these cultured-derived macrophages were adhered to C1q-coated surfaces. However, when cultured in a defined serum-free medium, these cells did not ingest EAC4b, even in the presence of C1q. To investigate this differential responsiveness, we studied the effects of C1q in conjunction with cell-activating agents on CR1 activation. Treatment of serum free cultured monocytes with phorbol dibutyrate (PDBu), prior to addition of the targets, induced these cells to ingest EAC4b. In addition, when exposed to C1q, both the percentage of these PDBu mononuclear phagocytes ingesting EAC4b and the number of targets ingested increased threefold over the level achieved by macrophages treated with PDBu alone. The chemoattractant N-formyl-methionyl leucyl-phenylalanine did not activate CR1-mediated phagocytosis and did not substitute for PDBu in causing synergy with C1q. Freshly isolated monocytes adhered to human serum albumin-coated glass slides in the absence or presence of PDBu did not phagocytose EAC4b. Also C1q did not stimulate monocyte CR1-mediated phagocytosis. However, addition of PDBu to cells adherent to the C1q surface triggered phagocytosis of EAC4b. The concentration of PDBu and the time of addition of PDBu relative to addition of the EAC4b targets were found to be important parameters for the achievement of maximal synergy in both the freshly isolated and cultured cell systems. This enhanced phagocytic activity was also seen with cells adhered to the purified collagen-like, pepsin-resistant, fragment of C1q. Since this region was previously shown to interact with C1q surface receptors, it appears that occupancy of this receptor is triggering events contributing to the enhanced cellular function. These experiments suggest that C1q and PDBu promote ingestion via CR1 by different but synergistic mechanisms. These data also demonstrate that the CR1-mediated enhancement of phagocytosis is not specific for FcR-mediated ingestion, but also applies to phagocytosis via CR1. PMID- 2975600 TI - Characterization of the human type I interferon receptor by ligand blotting. AB - The human type I interferon (IFN) receptor was characterized by ligand blotting. In this method, plasmalemma proteins or detergent-lysed whole-cell extracts from human Burkitt lymphoma cell lines were separated on polyacrylamide gels and subsequently transferred onto nitrocellulose sheets. Probing the blots with 3 x 10(-10) M 125I-labeled recombinant IFN-alpha A (125I-rIFN-alpha A) revealed an IFN-alpha-binding protein with an apparent molecular mass of 95 kDa (p95). Performing the electrophoretic run under reducing conditions completely abrogated the signal on the blot, indicating that the type I IFN receptor contains a disulfide bond essential for IFN binding. Optimal binding of 125I-rIFN-alpha A occurred at pH 9. The specificity of the binding reaction was established by simultaneously adding an excess of unlabeled IFN during incubation of the blots with 125I-rIFN-alpha A. The addition of either unlabeled IFN-alpha or IFN-beta, but not IFN-gamma, abolished the binding of 125I-rIFN-alpha A to p95. 125I-IFN gamma at 1.25 x 10(-11) M bound to two proteins distinct from p95 with apparent molecular mass of 92 and 87 kDa, respectively. Saturability of 125I-rIFN-alpha A binding was demonstrated by probing a constant amount of membrane proteins with increasing amounts of 125I-rIFN-alpha A. Scatchard analysis of the binding data yielded an apparent Kd of 5.4 x 10(-10) M for the immobilized type I IFN receptor. The expression of p95 on IFN-alpha-resistant and -sensitive cells was indistinguishable. We conclude that p95 is the IFN-alpha/beta receptor and that two proteins (p92 and p87) can specifically bind IFN-gamma. These results indicate that ligand blotting is a versatile method for characterization of unmodified IFN receptors and IFN-receptor interaction and could also provide a new investigational approach for other cytokine receptor systems. PMID- 2975601 TI - A novel human V delta gene expressed predominantly in the Ti gamma A fraction of gamma/delta+ peripheral lymphocytes. AB - We have characterized a functional T cell receptor (TcR) delta transcript in a Ti gamma A+ human cloned cell line derived from peripheral blood. This cDNA includes a novel V gene (V-AB12), whose expression was initially studied in a series of TcR gamma/delta+ clones. Nine Ti gamma A+ clones derived independently from distinct donors have been tested: each of them was found to possess a unique V AB12/J-IDP2 5.5-kb Eco RI rearrangement, which was constantly transcribed. Surface expression of the protein encoded by this unique rearranged gene was demonstrated by immunoprecipitations performed on three Ti gamma A+ polyclonal cell lines using a specific rabbit heteroantiserum. Further analysis strongly suggested that a monoclonal antibody (mAb), designated anti-BB3, detects a V-AB12 encoded antigenic determinant on the cell surface. Double-color immunofluorescence analysis of peripheral blood lymphocytes from ten donors indicated that most BB3+ cells are recognized by anti-Ti gamma A mAb. In previous studies, we have shown that a majority of TcR gamma/delta+ peripheral T cells expresses a gamma chain including V9 (Ti gamma A) and most frequently JP-encoded peptides. Given the present results on the delta chain, it can be concluded that, in many individuals, a predominant fraction (V gamma 9+/V-AB12+) of circulating CD3+ TcR alpha/beta- T lymphocytes expresses a receptor with little, if any, combinatorial diversity. PMID- 2975602 TI - Comparison of interleukin 2 and 12-O-tetradecanoylphorbol 13-acetate as signals for protein kinase C activation in purified human T lymphocytes. AB - Interleukin 2 (IL2) and 12-O-tetradecanoylphorbol 13-acetate (TPA) have been compared for their ability to induce translocation of protein kinase C (PKC) in T lymphocytes prestimulated with anti-CD3 monoclonal antibody (mAb), either in the presence or absence of monocytes. TPA alone did not promote purified T cell growth, but it was able to induce a transient, within 30 min, translocation of PKC activity. The profiles of PKC association with the membrane of the T cells under TPA stimulation were quite similar when either the anti-CD3 mAb or the fixed monocytes, or both, were added to the T cells. The decrease of cytosolic PKC under TPA stimulation was less pronounced for the purified T cells stimulated with anti-CD3 mAb, fixed monocytes alone or both than for unstimulated purified T cells. Even in the absence of monocytes, the addition of exogenous IL2 to the anti-CD3 mAb-treated T cells resulted in PKC translocation, with a transient increase in the PKC activity found in both the particulate and cytosolic fractions. When exogenous IL2 was added to the proliferating T cells, the association of PKC with the membrane was prolonged and the activity did not reach a plateau during the first 2 h after the IL2 stimulation. In parallel, the level of PKC associated with the membrane was higher in proliferating cells than in resting cells even 4 days after stimulation. These results suggest that activation of PKC by IL2 might be different from the direct activation of PKC by TPA and that a specific activation pathway, at least kinetically distinct from the classical phosphatidyl inositol diphosphate degradation by phospholipase C, might be involved during IL2 stimulation of T lymphocytes through high-affinity IL2 receptors. PMID- 2975603 TI - Functional and biochemical characterizations of avian T lymphocyte antigens identified by monoclonal antibodies. AB - Seven monoclonal antibodies (mAb) were used to characterize antigens present on chicken T lymphocytes and on natural killer cells by flow cytometry, radioimmunoprecipitation and by effects on cell-mediated cytotoxicity and mitogen induced proliferation. mAb CTLA8 and 5 stained 73% of thymus, 44% of spleen and 51% of peripheral blood lymphocytes (PBL), respectively, and immunoprecipitated 65- and 45-kDa proteins from detergent extracts of 125I surface-labeled thymocytes. Pretreatment of splenic lymphocytes with mAb CTLA5 and 8 in the presence of rabbit complement (C) eliminated the concanavalin A (Con A)-induced T cell proliferative responses. mAb CTLA3, 4 and 9 stained 43% of thymus, 36% of spleen and 18% of PBL, and immunoprecipitated 33-35-kDa proteins. Pretreatment of spleen cells with mAb 4 or 9 plus C reduced, but did not eliminate, the Con A induced proliferative response and significantly reduced both major histocompatibility complex (MHC)-restricted and non-MHC-restricted cellular cytotoxicity. mAb CTLA1 and 6 stained 58% of thymus, 13% of spleen and 19% of PBL. mAb CTLA1 and 6 immunoprecipitated a 65-kDa protein. mAb CTLA1 and 6 had no effect on the Con A-induced blastogenesis and CTLA6 caused no decrease in virus specific cytotoxic T lymphocyte and natural killer activity. These results indicate that (a) mAb CTLA5 and 8 identify antigens on mature T lymphocytes that are similar in tissue distribution, molecular mass and function to the mammalian CD5 antigen; (b) mAb CTLA3, 4 and 9 detect the avian homologue of CD8 antigen; and (c) mAb CTLA1 and 6 identify the avian homologue of CD4 antigen. PMID- 2975604 TI - Mechanism of skin allograft enhancement across an H-2 class I mutant difference. Evidence for involvement of veto cells. AB - Intravenous injection of spleen cells across mutant class I H-2 incompatibility results in a drastic donor-specific prolongation of skin allograft survival and a marked decrease in the donor-specific cytotoxic T lymphocyte precursor (CTLp) frequency. This immunosuppressive effect depends on the presence of radiosensitive T cells in the donor cell inoculum. It was excluded that a graft vs.-host reaction was responsible for the observed effects. In mixing experiments, spleen cells from animals transfused with allogeneic lymphocytes could not suppress a normal CTL response against the alloantigen, despite an excess of putative recipient-derived spleen suppressor cells. The data are compatible with the idea that donor T cells function as veto cells which inactivate recipient CTLp directed against the alloantigen expressed by the veto cell. PMID- 2975605 TI - Evidence for homogeneity of thromboxane A2 receptor using structurally different antagonists. AB - Nine structurally dissimilar thromboxane antagonists (SQ 29548, ICI 185282, AH 23848, BM 13505 (Daltroban), BM 13177 (Sulotroban), SK&F 88046, L-636499, L 640035 and a Bayer compound SK&F 47821) were studied for activity as thromboxane A2 receptor antagonists. The assays used were inhibition of responses induced by the thromboxane mimetic, U46619, on human washed platelet aggregation, rabbit platelet aggregation, rabbit aortic strip contraction, anaesthetised guinea-pig bronchoconstriction, and a radio-labelled ligand (125I-PTA-OH) binding assay as a measure of affinity for the human platelet receptor. The results of the present study, with activities spanning at least four orders of magnitude along with statistically significant correlations (at least P less than 0.01), strongly suggests that between assays, antagonists and species a homogenous population of thromboxane A2 receptors exists. This finding is in contrast to those of a close series of 13-azapinane antagonists studied by other workers which have suggested receptor heterogeneity. PMID- 2975606 TI - Hypotensive response to atrial natriuretic factor in conscious chronic pulmonary hypertensive rats. AB - The hypotensive effects of anaritide (Wy-47,663), a 26 amino acid atrial natriuretic factor, were examined in conscious, monocrotaline-induced chronic pulmonary hypertensive rats. Anaritide (0.25-8 micrograms/kg per min i.v.) decreased systemic arterial pressure at 2-8 micrograms/kg per min by as much as 15.6 +/- 0.4 mm Hg. The pulmonary hypotensive response to anaritide was not different from vehicle treatment. We conclude that anaritide is more effective in decreasing systemic than pulmonary arterial pressures in monocrotaline-induced pulmonary hypertensive rats. PMID- 2975607 TI - Endothelin stimulates atrial natriuretic peptide (ANP) release from rat atria. PMID- 2975608 TI - 8-Hydroxy-2-(di-n-propylamino)tetralin, a selective serotonin1A receptor agonist, reduces the immobility of rats in the forced swimming test by acting on the nucleus raphe dorsalis. AB - 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), a selective serotonin1A (5 HT1A) receptor agonist, was studied for its anti-immobility activity in the forced swimming test when administered into the raphe nuclei medianus and dorsalis of rats. At concentrations ranging from 0.5 to 5 micrograms, 8-OH-DPAT significantly reduced the immobility of rats when administered into the nucleus raphe dorsalis, but only 5 micrograms was effective when administered into the nucleus medianus. The activity of rats in an open-field under conditions identical to those used in the forced swimming test was not significantly changed by various concentrations of 8-OH-DPAT administered into the nucleus raphe dorsalis, but was significantly increased by an infusion of 5 micrograms 8-OH DPAT into the nucleus raphe medianus. The effect of an infusion of 1 micrograms 8 OH-DPAT into the nucleus dorsalis was prevented by infusing 2.5 micrograms (-) propranolol or 2.5 micrograms (-)-pindolol into the same area 5 min before 8-OH DPAT or by treating the animals with sulpiride systemically (100 mg/kg i.p.) or centrally (in the nucleus accumbens; 1 microgram/0.5 microliter). The results suggest that 8-OH-DPAT reduces the immobility of rats by activating dopamine transmission, probably in the nucleus accumbens, as a consequence of its ability to reduce the activity of 5-HT neurons that originate in the nucleus raphe dorsalis. In view of the similarities between the effects of well-established antidepressants and 8-OH-DPAT in the forced swimming test, it is suggested that 5 HT1A receptor agonists may constitute a novel class of antidepressant agents. PMID- 2975609 TI - Bronchoneutral effects in hypertensive asthmatics--celiprolol versus chlorthalidone. AB - The actions of celiprolol and chlorthalidone on lung function were studied in a randomized double-blind study of 30 patients. At the end of a 4-week placebo run in period each patient had a supine diastolic blood pressure between 90 and 115 mmHg, and a forced expiratory volume at the first second of expiration (FEV1) 50 80% of the predicted value in patients with bronchial asthma, and between 40% and 70% of the predicted value in patients with chronic airways obstruction. The active treatment period lasted 12 weeks during which patients received either celiprolol, 200-600 mg daily, or chlorthalidone 12.5-37.5 mg daily. Lung function, blood pressure and heart rate were measured and assessed every 2 weeks during active treatment. Systolic and diastolic blood pressures were reduced during treatment with both drugs, but FEV1, forced vital capacity and forced expiratory volume at 25-75% of vital capacity were not significantly altered. These results suggest that celiprolol does not adversely affect pulmonary function in hypertensive patients who also have chronic bronchial obstruction. PMID- 2975610 TI - Effect of long-term treatment with celiprolol on pulmonary function in a group of mild hypertensive asthmatics. AB - Previous studies have shown that single dose or short-term administration of celiprolol does not impair pulmonary function in patients with asthma. The long term effects of celiprolol and chlorthalidone on respiratory function in mild hypertensive asthmatic patients were compared. Following a 1-month placebo washout period, five patients were randomized to 3 months of treatment with celiprolol, and four patients were treated with chlorthalidone. This double-blind study was followed by a 12-month open trial in 13 asthmatics. Forced expiratory volume at the first second of expiration (FEV1) remained unchanged throughout the 12-month treatment period. This finding is of interest since a spontaneous decrease in FEV1 of 20 ml/year has been observed in recent surveys of asthmatic patients. No changes in peak expiratory flow rate or maximum expiratory flow at 50% of vital capacity were observed during the study, but measurements of FEV1 before and 2 h after drug administration indicated that celiprolol had no direct bronchodilating effect. The responsiveness to inhaled salbutamol was unaltered during the 12-month trial period, demonstrating that celiprolol does not block the beta 2-adrenoceptors on bronchial smooth muscle. Our results indicate that celiprolol is a safe long-term treatment for hypertension in asthmatic patients. Nevertheless, a survey of a large number of patients may be useful in order to detect individuals at risk, since sensitivity to beta-blockade may vary between patients. PMID- 2975611 TI - Vasodilating effects of celiprolol in patients with peripheral obliterative arterial disease. AB - Disturbances of peripheral blood supply have hitherto been considered contra indications to the use of beta-blockers. This is probably due to an initial increase in peripheral resistance seen with these agents. The question of whether beta-blockers should be used with caution, or not at all, in patients with peripheral arterial disease is of considerable clinical relevance, since coronary heart disease and hypertension often coexist with peripheral obliterative arterial disease. The influence of a single 200 mg dose of celiprolol on peripheral blood flow was studied in 17 male hypertensive patients (mean age: 61.4 years) with stage II peripheral vascular disease of the pelvis or upper leg. These patients were then treated with 200 mg/day celiprolol for a period of 6 weeks. Despite abstinence from physical training, the pain-free walking distance improved slightly during treatment, and the blood pressure and pulse rate were both reduced. Celiprolol did not adversely affect peripheral blood flow or clinical symptoms. This new beta-blocker, therefore, should not be contra indicated in patients with peripheral vascular disease, in the form of intermittent claudication. PMID- 2975613 TI - Voice fundamental frequency as an auditory supplement to the speechreading of sentences. AB - Recognition of words in conversational sentences of known topic was measured in nine normally hearing subjects by speechreading alone and by speechreading supplemented with auditory presentation of the output of an electroglottograph. Mean word recognition probability rose from 30% to 77% with the addition of the acoustic signal. When this signal was filtered to remove possible high-frequency spectral cues, the supplemented score fell, but only by a marginally significant 7 percentage points, supporting the conclusion that voice fundamental frequency was the principal source of enhancement. Enhancement occurred for all subjects, regardless of speechreading competence. PMID- 2975612 TI - 24-h ambulatory non-invasive blood pressure monitoring in the assessment of the antihypertensive action of celiprolol. AB - The well-known limitations of traditional sphygmomanometer blood pressure measurements have led to the development of a number of invasive and non-invasive 24-h blood pressure monitoring techniques which provide information on blood pressure in daily life. A non-invasive portable blood pressure monitor (IRC Spacelab S5300 monitor) was used to assess the antihypertensive action of celiprolol, 400 mg once daily, in a randomized, double-blind, crossover, placebo controlled study. After a washout period of 7 days, 15 patients with mild or moderate essential hypertension were subjected to three 24-h blood pressure recordings: at the end of washout, and after 1 month of placebo and celiprolol treatment. In the 10 responsive patients, celiprolol reduced 24-h systolic blood pressure, diastolic blood pressure and heart rate by 6.8%, 8.1% and 2.7%, respectively, when compared with placebo. The reduction in blood pressure was also evident during the night and the antihypertensive action of celiprolol was maintained 24-h after administration. Celiprolol had no effect on 24-h blood pressure or heart rate variability and, therefore, did not produce any alteration in cardiovascular homeostasis. These results demonstrate that celiprolol is an effective once daily treatment for hypertension. PMID- 2975614 TI - [Atrial natriuretic factor in cirrhosis]. PMID- 2975615 TI - Analysis of the promoter of the Rh2 opsin gene in Drosophila melanogaster. AB - We have analyzed the cis-acting regulatory sequences of the Drosophila melanogaster Rh2 gene that encodes the protein component of a rhodopsin which is expressed in ocellar photoreceptor cells. DNA fragments containing the start point of transcription of the Rh2 gene were fused to either the Escherichia coli chloramphenicol acetyltransferase (CAT) or lacZ (beta-galactosidase) genes and introduced into the Drosophila germline by P-element-mediated transformation. Expression of the E. coli genes was then used to assay the ability of various sequences from the Rh2 gene to confer upon the indicator genes the Rh2 pattern of expression. Fragments containing between 4.3 kb and 183 bp upstream of the start of transcription plus the first 32 bp of the 5'-untranslated leader were found to result in nearly identical levels of head-specific CAT expression. Deletion of Rh2 sequences distal to position -112 bp resulted in loss of detectable CAT expression from these Rh2/CAT fusion constructs. We have, therefore, defined a region essential for head-specific expression of the Rh2 gene to a region extending from -183 to -112. We have determined the DNA sequence of the Rh2 promoter from -448 to +32 and have found an 11-bp sequence which is also present in the upstream flanking sequences of two other photoreceptor-specific genes (ninaE and ninaC). By histochemical staining of beta-galactosidase expressed under the control of the Rh2 promoter and by analyzing the effect of the ocelliless mutation on the expression of an Rh2/CAT fusion gene, we have been able to demonstrate that this promoter is active in ocelli. PMID- 2975618 TI - [Current objectives of occupational hygiene and pathology in Georgian SSR]. PMID- 2975617 TI - Age-related decline in cell-mediated immunity in the C58 leukemic mouse strain. AB - The leukemia-prone C58 strain of mouse was examined for age-related changes in cellular immune function. Proliferative responses of lymphocytes to autologous and allogeneic stimulator cells [autologous mixed lymphocyte response (AMLR) and mixed lymphocyte response (MLR), respectively] and to mitogens were tested both prior to and around the usual age of disease onset which occurs at 7-8 months. Leukemia in these animals was defined by elevated peripheral blood and splenic white blood cell counts. The AMLR declined greater than 30% by 6-7 months of age and was virtually absent by 8 months of age even in animals that were not overtly leukemic. The MLR declined precipitously (greater than 95%) at 9 months of age. Both declines occurred at a younger age in C58 mice than in nonleukemic strains. Mixing experiments with cells from young and old animals indicate a defect in the Ly 1+23-, L3T4+ responding T cells. No evidence indicating a role for suppressor cell activity in this decline of cell-mediated immunity could be found. Deficiencies in cytokine (IL-2 and IL-1) production were not observed except in the oldest mice tested. Around the usual time of disease onset, splenic natural killer (NK) cell activity declines sharply even in nonleukemic mice. Cell-mixing experiments showed no evidence of suppressor cell activity by spleen cells from older mice, leukemic or nonleukemic, on the NK cell activity of young adult animals. Interferon alpha, beta treatment enhanced the NK activity of cells from old mice but did not restore the level of activity seen in young mice. Evidence has therefore been found for a premature decline in cellular immune function in two responses with proposed immunoregulatory roles, the AMLR and NK cell activity. It is possible that their decline could play a predisposing role in the onset of this retroviral leukemia or that these cell populations may be the target of the retrovirus. PMID- 2975616 TI - Chain-bias of Escherichia coli Rec-mediated lambda patch recombinants is independent of the orientation of lambda cos. AB - Chi is a hotspot for homologous recombination mediated by the RecBCD (Rec) pathway of Escherichia coli. For Rec-mediated recombination of phage lambda, the orientation of lambda cos in the lambda chromosome dictates the direction of travel of RecBCD enzyme through DNA and dictates which orientation of Chi or Chi like sequences will be active in stimulating recombination. I previously found that Rec-mediated lambda patch heteroduplexes, stimulated by Chi or not, are chain-biased; at the lambda P locus, recombinant information resides on the lambda r chain. This bias exists in the presence or absence of Chi sites. Reported herein is the finding that r-chain-bias at the P locus is independent of the orientation of lambda cos and thus also independent of the orientation of active Chi's or Chi-like sequences and of the direction of travel of RecBCD enzyme. These results disprove previously elaborated models in which a chain specific nick at Chi initiates recombination, and imply that some other chain distinguishing process is involved with recombination. Replication and transcription are candidates for such a process. PMID- 2975619 TI - [Sanitary-hygienic characteristics of air environment and microflora at a cotton processing plant]. PMID- 2975620 TI - [Effect of occupational factors on the condition of the oral cavity in workers engaged in the processing of non-ferrous metals]. PMID- 2975621 TI - [Decreasing noise level during bottling of liquid foods]. PMID- 2975622 TI - [Status of regional hemodynamics of upper extremities in female tea leaf pickers]. PMID- 2975623 TI - [Characteristics of the structure of a solid phase of ore aerosol during the operation of power-driven Diesel equipment]. PMID- 2975624 TI - [Complex evaluation of the dust factor in the analysis of the indicators of the incidence of pneumoconiosis in miners]. PMID- 2975626 TI - Distal oblique osteotomy for intractable plantar keratosis of the middle three metatarsals. AB - On the middle metatarsals of 49 patients with symptomatic intractable plantar keratosis, 69 distal oblique osteotomies were performed. With an average follow up time of 16 months, 83% of the results were rated good. This procedure is contraindicated for an intractable plantar keratosis with an associated fixed metatarsphalangeal joint deformity, however. PMID- 2975625 TI - [General pharmacological studies on a bronchodilator, oxitropium bromide (Ba 253)]. AB - The effects of oxitropium bromide (Ba 253) on respiratory, cardiovascular, digestive and urogenital systems were studied. Ba 253 increased heart rate in anesthetized cats at low doses (0.1-0.3 mg/kg, i.v.) and decreased blood pressure in dogs and cats at high dose (3 mg/kg, i.v.). Aerosol inhalation of a high concentration of Ba 253, however, did not influence the heart rate. Ba 253 enhanced the isoproterenol-induced inotropic action and vasodilation. Intestinal transport of mice were inhibited by Ba 253 (s.c.), but not inhibited by oral administration. Ba 253 (1-30 mg/kg, i.v.) enhanced the motility of rat uterus in vivo, but inhalation of the Ba 253 aerosol did not have any affect. Ba 253 had no effects on vasoconstriction, spontaneous motility of the ileum, bile secretion, urinary excretion and spontaneous motility of the urinary bladder. These results indicate that intravenous or subcutaneous administration of Ba 253 decreased blood pressure, enhanced uterus motility and inhibited intestinal transport, but the inhalation or oral administration, even at high doses, has no effects on the cardiovascular system and uterine motility. PMID- 2975627 TI - [Bronchodilator effectiveness of theophylline]. PMID- 2975628 TI - The effect of repetitive haemorrhage on plasma cortisol, beta-endorphin and N terminal pro-opiomelanocortin in conscious sheep. AB - We measured the effect of repeated haemorrhagic stress, performed on four consecutive days in conscious adult sheep, on the plasma concentrations of cortisol and ACTH-related peptides to determine whether the pituitary-adrenal response was altered by stress repetition. Peptides from the C-terminus of the ACTH pro-hormone was measured by beta-endorphin RIA. Glycopeptides derived from the N-terminus of the ACTH pro-hormone were measured by tau 3-MSH RIA. The immunoreactive tau 3-MSH in sheep plasma was found to have an apparent molecular weight of approximately 10,000 by gel chromatography through Sephadex G-75, which is similar to the size of the major circulating form of pro-tau-MSH found in human and rat plasma. Daily haemorrhage consistently elevated plasma concentrations of cortisol and pro-tau-MSH. There was no significant difference in the daily responses of either cortisol or pro-tau-MSH when considered individually. However, there was a significant change over the four days in the relationship between the cortisol and pro-tau-MSH responses, as judged by analysis of variance of the difference in daily z-scores of cortisol and pro-tau MSH. This trend indicated a relative increase in the secretion of pro-tau-MSH from the pituitary compared to the cortisol response, and suggested that repeated exposure to stressful stimuli may alter the pituitary-adrenal-axis. PMID- 2975629 TI - Metabolic effects of combined cyproterone acetate and percutaneous 17 beta oestradiol after six and twelve months therapy in 61 patients. AB - The metabolic effects of combined cyproterone acetate (50 mg) and percutaneous 17 beta oestradiol were studied during one year in 61 patients admitted for hyperandrogenia. Before treatment and at 6 and 12 months the following tests were performed: oral glucose tolerance test (OGTT) with insulinemia dosage, determination of total cholesterol, LDL, VLDL and HDL fractions, triglycerids, A1 and B apoproteins, liver function tests: bilirubinemia, alkaline phosphatases, transaminases and gamma glutamyl transferases. The patients' mean age was 27.0 +/ 6.8 years, the body mass index was 22.4 +/- 3.5 kg/m2. After one year of treatment the body mass index was not modified. Blood glucose slightly increased during OGTT; at 6 months this was significant at +30 minutes, at 12 months at +30, +60 and +90 minutes (P less than 0.05). There was no variation in insulinemia during OGTT. Total cholesterol decreased significantly at 6 and 12 months (P less than 0.001), this was associated with a decrease in HDL cholesterol, but without modification of the LDL + VLDL/HDL ratio. Decrease in HDL cholesterol was associated with a significant decrease in A1 apoproteins. No change in triglycerids and in liver function tests was observed at either date. In conclusion the metabolic effects of this association are described. These effects are minimal compared to those observed with cyproterone acetate and ethinyl oestradiol association in the literature. However, attention should be drawn to the possibility of glucose intolerance and decrease in HDL cholesterol and A1 apoproteins. PMID- 2975630 TI - Effect of diclofenac on plasma levels of immunoreactive prolactin, follicle stimulating hormone, luteinizing hormone, thyrotropin, and beta-endorphin in humans. AB - Prostaglandins have been shown to modulate the secretion of several pituitary hormones, suggesting that therapeutic doses of nonsteroidal anti-inflammatory drugs may change basal hormone levels. In this study, plasma levels of prolactin, follicle stimulating hormone, luteinizing hormone, thyrotropin and beta-endorphin were determined in 6 healthy men after administration of diclofenac, a prostaglandin synthesis inhibitor. The subjects were given 75 mg intramuscularly and 50 mg orally at 08.00 h the first day, 50 mg orally at 08.00, 12.00 and 20.00 h the second day and an additional 50 mg orally at 08.00 h the third day. Blood samples were collected throughout these 3 days. Diclofenac resulted in a significant and sustained decrease in plasma level of prolactin (p less than 0.005). The other hormones did not demonstrate significant change following diclofenac administration. These data suggest that administration of a prostaglandin synthesis inhibitor, such as diclofenac, selectively alters basal pituitary secretion of prolactin in humans without a detectable effect on plasma levels of other pituitary hormones. This study supports the hypothesis that prostaglandins are necessary for maintaining basal level of prolactin secretion in man. PMID- 2975631 TI - Sustained impairment of pituitary and testicular function in prostatic cancer patients treated with a depot form of a GnRH agonist. AB - Seven patients suffering from prostatic cancer were treated with a slow-release D Trp-6-LHRH preparation for a period of 24-32 months. LH, FSH, PRL and testosterone levels were evaluated before and at the end of treatment and then 40 days later. Baseline and GnRH-, TRH-, and HCG-stimulated hormonal values decreased after treatment. The possibility that a long-term treatment with GnRH analogues induces a sustained suppression of pituitary and testicular function is suggested. PMID- 2975632 TI - Expression of 67,000 MW calcimedin and its binding protein in resident and thioglycolate-elicited macrophages. AB - Calcimedins are newly recognized calcium-dependent hydrophobic-binding proteins. The 67,000 MW calcimedin exhibits calcium-dependent specific binding to a 60,000 MW intracellular acceptor protein (60,000 MW). Macrophages possessed 30,000 MW and 67,000 MW calcimedins and 60,000 MW acceptor protein, identified by immunofluorescence and confirmed by SDS-PAGE and Western immunoblots. Compared with low levels in resident macrophages, thioglycolate-elicited inflammatory macrophages showed large increases in the levels of 67,000 MW calcimedin and 60,000 MW acceptor protein, especially at cell-limiting membranes. Levels of calmodulin and 30,000 MW calcimedin appeared similar in both populations. Trypsinization removed most 67,000 MW calcimedin and some 60,000 MW acceptor protein staining. The calcimedins appear to represent novel macrophage differentiation antigens. Their modulation suggests that they may play a role in the inflammatory response. PMID- 2975633 TI - Characterization of antigen-presenting activity of intestinal mononuclear cells isolated from normal and inflammatory bowel disease colon and ileum. AB - Antigen-presenting activity in mononuclear cells, isolated from normal and inflamed human ileum and colon, has been characterized using allogeneic mixed lymphocyte reaction with resting T cells as responders. Greatest proliferation was induced by fibronectin-adherent (macrophage-enriched) cells, and least by fibronectin non-adherent (macrophage-depleted) cells and by mononuclear cells depleted of macrophages by panning with monoclonal antibody 3C10. When intestinal mononuclear cells and allogeneic T cells were incubated in large numbers, clusters were observed. These clusters contained cells with a dendritic morphology that were strongly HLA-D-positive and which also stained with macrophage-specific monoclonal antibodies 3C10, EMB11 and Y1/82A. These cells were closely associated with proliferating T cells. Studies comparing mononuclear cells isolated from normal and inflamed colonic mucosa suggest that the latter may have enhanced antigen-presenting capacity. PMID- 2975635 TI - Effect of Ly 5 allotype on in vitro immune responses. AB - We have tested in vitro immune responses of several kinds to determine if Ly 5 allotype influences reactivity of murine splenocytes in processes thought to involve the T200 glycoprotein. Matings were established among C57BL/6J (B6) (Ly 5.1) and C57BL/6-Ly 5.2 (B6-Ly 5.2) congenic mice (both H-2b) to obtain sibling mice segregating for alloalleles of the Ly 5 system. F2 progeny of the three Ly 5 genotypes were tested for antibody-dependent cell-mediated cytotoxicity (ADCC), proliferation in allogeneic mixed lymphocyte culture (MLC), mitogen responsiveness, natural killer (NK) cell activity, and cytotoxic T-lymphocyte (CTL)-mediated cytotoxic activity. We observed that in Ly 5 segregant mice, higher CTL activity was associated with the Ly 5.2 type. No allotype effect was observed in MLC, mitogen responses, and NK cell-mediated cytolysis. In the parental and F1 animals, mice carrying the Ly 5.2 allele had significantly higher ADCC levels, though this effect was not seen in the segregants. Our results indicate that Ly 5 or some closely linked gene or genes influence CTL activity of murine splenocytes in vitro. PMID- 2975634 TI - Functional evidence for a monoclonal antibody that binds to the human IL-4 receptor. AB - The complex pleiotropic effects of the T-cell derived lymphokine interleukin-4 (IL-4) are becoming increasingly well documented; however, functional studies have been hampered by the lack of reagents directed against the receptor for this factor. In this report, we present data which suggest that the monoclonal antibody MR6 binds to the human interleukin-4 receptor (IL-4R). Addition of MR6 to cultures of T cells proliferating in response to IL-4 inhibited this response in a dose-dependent fashion, giving total inhibition at 10 micrograms/ml. Similarly, the IL-4-dependent production of specific antigen-induced IgE by B cell populations was completely abrogated by MR6. Flow cytometric studies of the modulation of cell surface molecules after T-cell activation suggest that expression of the molecule detected by MR6 (p145-MR6) correlates inversely with that of the interleukin-2 receptor (IL-2R). These data, together with the previously determined molecular weight and tissue distribution of this molecule, strongly indicate that MR6 binds to the human IL-4R. PMID- 2975636 TI - Selenium status in females with occupational cervico-brachial complaints. AB - A hypothesis that selenium deficiency predisposes the development of occupational cervicobrachial complaints was tested in 134 females working in an assembly factory, with constrained work postures and repetitive work tasks. Seventeen subjects, who reported intake of selenium tablets, had higher plasma selenium levels than the others (104 vs 89 micrograms/l, P = 0.01). Among those who did not take selenium tablets, 21% reported symptoms from the upper back during the last 7d, 21% from the neck, 38% from the shoulders, 15% from the elbows, and 27% from the hands. Subjects with pain in their elbows had slightly, but significantly, lower plasma selenium levels than asymptomatics (84 vs 90 micrograms/l, P = 0.048). For the other anatomical regions, there were no statistically significant differences. Thus, there was no major association between selenium status and pain; conclusions regarding any minor association must await further studies. PMID- 2975638 TI - Urticarial vasculitis in adults. PMID- 2975639 TI - Lumen phase specific cross immunity between Hymenolepis microstoma and H. nana in mice. PMID- 2975637 TI - Incidence of spontaneous and provoked coronary spasms after percutaneous transluminal coronary angioplasty. AB - We studied the incidence of spontaneous and ergonovine-induced coronary arterial spasm during repeat coronary angiography in 96 consecutive patients with single vessel disease who had undergone successful angioplasty. Follow-up angiography was performed after a mean of 6 months (1-8 months). Sixty patients demonstrated no restenosis and in 36 patients restenosis (greater than 50% restenosis) occurred. Spasms of the arteries at the site of dilatation were significantly (P less than 0.001) more frequent in patients with restenosis (18/36; 50%) than in patients without restenosis (4/60; 7%). Before angioplasty, no differences were found in the clinical characteristics between the two groups. Likewise, the morphologic results of angioplasty were identical. Despite long-term treatment with nifedipine (30-60 mg daily) and aspirin (0.5 g daily), 14 of 18 patients with restenosis and coronary spasm suffered from spontaneous angina, as compared to only 3 of 18 patients with restenosis without demonstrable spasm. Three of the 4 patients without restenosis but with detectable spasm were also symptomatic. Thus our findings suggest that spasm of the coronary arteries achieves some importance as a pathophysiological factor for recurrence following coronary angioplasty. PMID- 2975640 TI - Synthesis and activity of partial retro-inverso modified atrial natriuretic factor analogs. AB - The synthesis and pharmacological activity of partial retro-inverso modified rat atrial natriuretic factor (rANF) analogs is described. The route to these compounds utilized a combination of solution and solid-phase methods. The analogs prepared all contain a reversed amide bond (psi[NHCO]) at the Ser 25 to Phe26 linkage. This bond has been suggested to play a key role in the metabolic inactivation of ANF. The analogs are of comparable potency to the endogenous peptide rANF1-28 in binding to cultured rat vascular smooth muscle cells, in relaxing serotonin contracted rabbit aortic rings, and as natriuretic/diuretic agents in anesthetized rats. None of the peptides has an extended duration of action in vivo. PMID- 2975641 TI - Depo-provera in the First World. PMID- 2975642 TI - [Photoallergic and allergic contact eczema caused by dibenzoylmethane compounds and other sunscreening agents]. AB - Dibenzoylmethanes are widely used in Europe in UV-A-absorbing sunscreens. We report on 21 patients with (photo)allergic contact dermatitis from 4-isopropyl dibenzoylmethane. Six subjects revealed, in addition, (photo)allergic reactions to 4-tert.butyl-4'-methoxy-dibenzoylmethane, seven to 3-(4-methylbenzylidene) camphor, two to 2-phenyl-benzimidazole-5-sulfonic acid and one to 2-phenyl-5 methylbenzoxazole. Allergic contact dermatitis from 2-phenyl-benzimidazole-5 sulfonic acid has not been reported previously. The relevance of sunscreens as a cause of (photo-)allergic contact dermatitis is discussed. PMID- 2975643 TI - [Symmetrical lividity of the fingers]. AB - Symmetric lividity of the soles of the feet was first reported in two children in 1925 by Pernet. The characteristic manifestation of this dermatosis consisted in hyperkeratosis and hyperhidrosis with livid discoloration of the pressure areas of the soles. Later the same name was applied to a similar dermatosis in which the hyperkeratotic and hyperhidrotic patches of skin on the soles had a whitish grey discoloration and the livid color, if present at all, was seen only over the marginal areas not affected by the keratosis. Similar livid keratoses affecting the palmar sides of the fingers have been seen only occasionally. The 17-year-old girl presented in this paper had a 11-year history of emotional hyperhidrosis and is a rare illustration of symmetrical lividity in its original form, localized to the fingers only. PMID- 2975644 TI - [Hailey-Hailey disease with dissemination and eczema herpeticatum in therapy with etretinate]. AB - We report a 39-year-old patient with Hailey-Hailey disease localized mainly in the genito-femoral and scrotal areas. During hospital treatment the condition worsened due to superinfection with Candida albicans and gram-negative bacteria. Treatment with systemic steroids followed by aromatic retinoids was tried: eczema herpeticum developed in the affected areas, and dissemination of Hailey-Hailey disease ultimately occurred with papulo-vesicular and histologically characteristic lesions. PMID- 2975645 TI - Electromyographic and skin resistance responses to osteopathic manipulative treatment for low-back pain. PMID- 2975646 TI - Pustular dermatitis caused by subcutaneous dracunculiasis in snakes. PMID- 2975647 TI - The legal obligation to learning-disabled and handicapped allied health students. AB - Recent legislation and judicial decisions regarding the legal responsibilities of postsecondary institutions to handicapped and learning-disabled students is discussed. The relevance of these decisions to administrators of allied health programs in the selection and retention of students becomes apparent as one reviews the increased number of litigations in the past few years. In summary, administrators must make reasonable accommodations to "otherwise qualified" handicapped individuals. They need not, however, accept students that could not benefit from the principal education given or would not likely pass certification exams in their field of study. PMID- 2975648 TI - Recent advances in the treatment of malignant ventricular arrhythmias: new hope for prolonged survival. PMID- 2975650 TI - Frostbite. PMID- 2975649 TI - Urinary incontinence in the elderly. PMID- 2975651 TI - Biologic effects of alcohol. PMID- 2975652 TI - The steady state intermediate of scallop smooth muscle myosin ATPase and effect of light chain phosphorylation. A molecular mechanism for catch contraction. AB - The ATP-induced difference UV-absorption spectrum of myosin isolated from the opaque portion of scallop smooth muscle (opaque myosin) was Ca2+-sensitive at 40 mM KCl and 1.5 M sucrose. On adding sucrose to 1.5 M, the turbidity of myosin decreased to 24% and the characteristic two forms of the difference spectrum, the ATP-form and ADP-form (Morita, F. (1967) J. Biol. Chem. 242, 4501-4506), were distinguishable. In the presence of Ca2+, the difference spectrum was the ATP form first and then decayed into the ADP-form with the depletion of ATP. In the absence of Ca2+, however, only the ADP-form was observed. The ADP-form observed in the absence of Ca2+ returned to the ATP-form when the regulatory light chain-a (RLC-a), one of the regulatory light chains of opaque myosin, was phosphorylated. These results suggest that the main intermediate at the steady state of opaque myosin ATPase is converted depending on the concentration of Ca2+, from EPADP in the presence of Ca2+ to EADP in the absence of Ca2+. It changes to EPADP in the absence of Ca2+ on the phosphorylation of RLC-a. Consistent results were obtained by measuring the ATP-induced Trp-fluorescence increase of opaque myosin in the absence of sucrose. Since the opaque portion of scallop smooth muscle is known to be responsible for catch contraction (Ruegg, J.C. (1961) Proc. R. Soc. London Ser. B 154, 224-249), these findings lead us to suppose that the opaque myosin in vivo may stay in the E.ADP complex during the catch state. It changes to EPADP by the phosphorylation of RLC-a, which may terminate the catch state. PMID- 2975653 TI - Crystallization of and preliminary crystallographic data for Bacillus stearothermophilus cyclodextrin glucanotransferase. AB - Cyclodextrin glucanotransferase from Bacillus stearothermophilus TC-91 has been crystallized from an ammonium sulfate solution by the dialysis equilibrium method. The crystals belong to the orthorhombic system, space group P2(1)2(1)2(1), with cell dimensions of a = 125.5 A, b = 88.1 A, and c = 81.5 A. The crystals appear to be suitable for X-ray structure analysis, diffracting to at least 2.1 A and being resistant to radiation damage. PMID- 2975654 TI - Effect of substitution of troponin C in cardiac myofibrils with skeletal troponin C or calmodulin on the Ca2+- and Sr2+-sensitive ATPase activity. AB - Troponin C was removed almost completely from the porcine cardiac myofibrils by the same extraction procedure using CDTA as that previously reported for the rabbit skeletal myofibrils (Morimoto, S. & Ohtsuki, I. (1987) J. Biochem. 101, 291-301), and the effects of substitution of troponin C in cardiac myofibrils with rabbit skeletal troponin C or bovine brain calmodulin were examined. While the ATPase activity of intact cardiac myofibrils or cardiac troponin C reconstituted cardiac myofibrils was activated at only a little higher concentration of Sr2+ than Ca2+, the skeletal troponin C-substituted cardiac myofibrils, as well as intact rabbit skeletal myofibrils, required more than 10 times higher concentration of Sr2+ than Ca2+ for activation of the myofibrillar ATPase activity. However, the concentrations of Ca2+ and Sr2+ required for the activation of the ATPase activity of the skeletal troponin C-substituted cardiac myofibrils were both about 5 times higher than those of intact skeletal myofibrils. The skeletal troponin C-substituted cardiac myofibrils, as well as intact skeletal myofibrils, also showed higher cooperativity in the Ca2+ activation of the ATPase activity than intact or cardiac troponin C-reconstituted cardiac myofibrils. The ATPase activity of calmodulin-substituted cardiac myofibrils was activated at a several times lower concentration of Ca2+ or Sr2+ than that of calmodulin-substituted skeletal myofibrils, while the ratios of the concentration of Sr2+ to Ca2+ required for activation were almost the same in both cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975656 TI - The Early Treatment Diabetic Retinopathy Study: an update. PMID- 2975655 TI - Mechanism of inhibition by alloxan of ATP-driven calcium transport by vascular smooth muscle microsomes. AB - The direct in vitro effects of alloxan on the Ca2+ handling by microsomal membranes isolated from dog mesenteric arteries were investigated. Preincubation of the vascular muscle microsomal membranes with alloxan showed a suppressive effect on both binding of Ca2+ (in the absence of ATP) and ATP-driven Ca2+ transport. Such an inhibition was time dependent, dose dependent, and temperature dependent. ATP-driven Ca2+ transport was much more susceptible to the inhibitory action of alloxan than Ca2+ binding under all experimental conditions examined. Alloxan inhibited ATP-driven Ca2+ transport at a comparable level over the entire period of Ca2+ uptake, but had no significant effect on the efflux of Ca2+ from preloaded microsomal membranes. This suggests that alloxan exerts its inhibitory effect on the ATP-driven Ca2+ transport via its action on the Ca-pump protein rather than the membrane permeability to Ca2+. Catalase and mannitol but not superoxide dismutase partially protected against such as inhibition by alloxan. The possible involvement of H2O2 mediating the inhibitory action of alloxan was further supported by the finding of a similar in vitro inhibitory effect of H2O2 on the ATP-driven Ca2+ transport by the vascular smooth muscle microsomes. PMID- 2975657 TI - Transplantation of immunologically modified pancreatic tissue. AB - The activation of an alloantigen reactive T cell requires alloantigen recognition and additional external signals provided by antigen presenting cells of host or donor antigen. Graft passenger leukocytes are capable of presenting alloantigen directly to recipient T cells and providing interleukin-1. Alternatively, shed alloantigen may be presented passively by recipient antigen presenting cells. Nearly all attempts to alter graft tissue immunogenicity prior to transplantation have focused on removing passenger cells. Attempts to modulate islet graft immunogenicity include conventional organ culture, high oxygen/high pressure culture, interim hosting, and treatment with ultraviolet irradiation, monoclonal antibodies and complement, and immunotoxins. While these techniques have been successful in rodent models, attempts to apply these methods to human islet transplants have been unsuccessful. PMID- 2975658 TI - Diabetic retinopathy: I. The measurement of intercapillary area in normal retinal angiograms. AB - A method for the quantitation of areas of non-perfusion of retinal angiograms is described. The values for the area of the foveal avascular zone (FAZ) and the intercapillary areas in the perifoveal region of ten angiograms of normal subjects were determined. The reproducibility of measurement between and within observers was good, correlation coefficients being 0.9963 and 0.9945, respectively, for the FAZ. The inter- and intra-observer coefficients of correlation for the perifoveal intercapillary areas were 0.9284 and 0.8687, respectively. The upper limit of normal intercapillary area was determined to be 0.07 mm2 in the first degree from the FAZ, 0.04 mm2 in the second degree and 0.025 mm2 elsewhere in the macular field. PMID- 2975659 TI - Diabetic retinopathy: II. Assessment of severity and progression from fluorescein angiograms. AB - A quantitative method for assessing the severity of retinopathy from fluorescein angiograms was applied to a group of patients starting continuous subcutaneous insulin infusion. There was a highly significant correlation between the area of nonperfusion as measured in the 20 degrees field centered on the right macula, and the mean retinopathy score derived by a standard method from a color retinal photographic survey. In those subjects who deteriorated more, there was an increase in the nonperfused areas, whereas the foveal avascular zone remained unchanged. PMID- 2975660 TI - Pancreatic transplantation: the Cambridge experience. AB - In contrast to heterotopic pancreas transplantation and conventional insulin therapy the paratopic positioning of a segmental pancreas graft provides physiological endocrine hormone delivery into the portal venous circulation. Metabolic studies performed on previously insulin dependent diabetic patients with renal failure with functioning paratopic pancreas transplants and heterotopic kidney grafts from the same donor showed near normal day to day glucose control with normal fasting glucose levels and normal HbA1 values. Hyperinsulinaemia was not seen in these patients. The effect of denervation of the pancreatic graft on the entero-insular axis and its influence on the islets were investigated. The incretin effect was preserved indicating a significant hormonal stimulation of the entero-insular axis. PMID- 2975661 TI - Pancreas transplantation: the Minnesota experience. AB - Between July 1978 and April 1987, a total of 182 pancreas transplants were performed at the University of Minnesota. For the first 100 cases (through October 1984), a variety of surgical techniques and immunosuppressive regimens were used, and 1 year patient and graft functional (insulin-independent) survival rates were 88% and 27%, respectively. From November 1984 to April 1987, a triple therapeutic drug regimen of cyclosporine, azathioprine, and prednisone was used for maintenance immunosuppression, and bladder drainage (BD) (n = 39; 38 cadaver (CAD) and 1 related (REL) donor grafts) and enteric drainage (ED) (n = 40; 21 CAD and 19 REL donor grafts) techniques were compared in 59 nonuremic, nonkidney (NUNK) transplant recipients, 21 recipients of previous kidney (PK) transplants and 8 uremic recipients of simultaneous pancreas and kidney (SPK) transplants. The survival rates were higher in recipients of BD CAD and ED REL than of ED CAD grafts (58% and 59% versus 29% at one year for all, and 84%, 84% and 40% for technically successful cases), but patient survival rates were similar (90%, 93% and 90% at one year). BD allows for early diagnosis of rejection based on urine amylase monitoring, and REL grafts are less prone to incite rejection; thus, we are currently performing only BD for grafts from CAD donors, while both techniques are used for REL donor grafts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975662 TI - Coping with autonomic neuropathy. AB - Diabetic autonomic neuropathy, the most common of all the complications associated with diabetes mellitus, is manifested as cardiovascular or gastrointestinal denervation, counterregulatory hormone dysfunction, and genitourinary tract denervation. Its treatment at this time is largely palliative, although there is some indication that control of hyperglycemia, hypertension, and obesity may prevent or delay its development. Both counseling and pharmaceutical approaches are essential to permit the afflicted diabetic patient a reasonably good quality of life. PMID- 2975663 TI - Diabetic cystopathy. AB - Diabetic cystopathy refers to the spectrum of voiding dysfunction in patients with diabetes mellitus. Diabetic cystopathy is marked by insidious onset and progression with minimal symptomology. The most common urodynamic findings are impairment of bladder sensation, increased post-void residual volume, decreased detrusor contractility that may progress to detrusor areflexia and diminished urinary flow. Treatment of diabetic cystopathy may be complicated by frequently occurring coexisting urologic conditions. The most common of these is bladder outlet obstruction. Therefore, treatment of diabetic cystopathy should be tailored to the symptom complex and clinical condition of the patient. Asymptomatic patients with manifestations of diabetic cystopathy may be treated with timed voiding. In contrast, the sine qua non for therapy in symptomatic patients is clean intermittent catheterization. While other modalities such as pharmacologic and surgical intervention have been described, none have been consistently effective. PMID- 2975664 TI - Compliance with chronic disease regimens: diabetes. AB - Patient compliance with the prescribed management regimen is presented as the fourth component responsible for control of blood sugar in insulin-taking diabetics, along with insulin dose, food consumed, and exercise performed. Noncompliance is ascribed to a large variety of reasons which vary in importance from diabetic to diabetic. A survey of ten diabetic women, two of whom were uremic and eight of whom were renal transplant recipients, was conducted to assess their compliance with advised shoe prescriptions. Observed compliance with wearing acceptable shoes is delineated as occurring because it fits with patients' life-styles. It was found that three of ten women refused to wear recommended footgear all of the time, whereas one woman varied her acceptance of advised shoes. Contracts between patients and health care team members are offered as a means of encouraging compliance; individualizing the health team's care of the patient is put forward as the most significant approach. PMID- 2975666 TI - Transplantation, hemodialysis, and continuous ambulatory peritoneal dialysis for end-stage renal disease in diabetic patients. PMID- 2975667 TI - Renal transplantation in diabetic patients is confirmed therapy while pancreas transplantation should be performed only in an investigational setting. AB - Kidney transplantation is now firmly established as the standard treatment for all diabetic patients with end-stage renal failure. In an analysis of all renal transplants at the University of Minnesota between June 1, 1980 and May 31, 1987, there were no differences in renal allograft functional survival rates for diabetic and nondiabetic recipients. At one year the survival rates were 84% (n = 151) and 86% (n = 260) for those treated with azathioprine, prednisone and ALG; 86% (n = 101) and 87% (n = 104) for those treated with cyclosporine-prednisone, and 92% (n = 165) and 89% (n = 191) for those treated with triple therapy (cyclosporine, azathioprine, and prednisone). Pancreas transplantation remains an investigational procedure for nonuremic diabetic patients but may be considered therapeutic in diabetic renal allograft recipients because such patients are obligated to immunosuppression and only the surgical risks of pancreas transplantation need to be considered, which are now acceptably low. Recipients of pancreas transplants performed simultaneous with the kidney have patient and pancreas graft survival rates of greater than 90% and +/- 60% at several institutions, including our own. The potential for benefit of pancreas transplantation, however, is greater in the nonuremic nonkidney transplant patient, and pancreas transplantations are being performed in such patients at a few institutions. An early beneficial effect of pancreas transplantation preexisting proliferative retinopathy has not been discerned, although long-term retinopathy has been stable in patients with functioning grafts. Preliminary studies have shown a beneficial effect on neuropathy and on microscopic lesions of diabetic nephropathy, but at the expense of cyclosporine toxicity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975665 TI - Preserving feet in the uremic diabetic: the physician's role. AB - We report the results of a study of 40 consecutive conditions seen in 38 ambulatory diabetics at a municipal hospital podiatry clinic and a university hospital nephrology clinic. The vast majority (92.5%) were preventable, had appropriate foot care procedures been followed. It is suggested that the lack of emphasis on prevention of foot pathology in diabetics by criteria for fee-for service payment in medicine leads to excessive and unnecessary limb morbidity. PMID- 2975668 TI - Rapid method for the analysis of red blood cell fatty acids by reversed-phase high-performance liquid chromatography. AB - A high-performance liquid chromatographic method was developed for a rapid qualitative and quantitative analysis of p-bromophenacyl esters of red blood cell fatty acids in humans. Both free and bound fatty acids, extracted with hexane-2 propanol (3:2) from packed red blood cells were derivatized with p-bromophenacyl bromide and analysed. Ten identical samples taken from a mixed pool of packed red blood cells from healthy subjects were analysed on two different columns. The fatty acid p-bromophenacyl esters were analysed on a 10 RP-18 column with methanol-acetonitrile-0.01 M ammonium formate as mobile phase and also on a 10 RP 8 column with acetonitrile-0.01 M ammonium formate as mobile phase. The two methods gave analogous results except in total analysis time: that on a 10 RP-8 column is ca. 40% shorter. Furthermore, a quantitative analysis of a standard solution to evaluate the extraction procedure in the absence or in the presence of the red blood cell core indicated a significant difference when the core is present. PMID- 2975670 TI - Antibody-dependent cellular cytotoxicity (ADCC)-mediated destruction of human immunodeficiency virus (HIV)-coated CD4+ T lymphocytes by acquired immunodeficiency syndrome (AIDS) effector cells. AB - The acquired immunodeficiency syndrome (AIDS) is defined in clinical terms by the development of Kaposi's sarcoma and/or severe opportunistic infections in persons without predisposing conditions. A hallmark of the syndrome has been a decrease in the number of CD4+ T helper cells. The reduction in the frequency of the CD4+ lymphocytes has been postulated to be primarily the result of human immunodeficiency virus (HIV) tropism and cytophathogenicity for the T-cell subset. Yet only a small percentage of cells in actually infected with HIV. Recently, we provided evidence indicating that AIDS patients' natural killer cells can mediate normal levels of antibody-dependent cellular cytotoxicity (ADCC) despite exhibiting a defect in natural killer (NK) effector function (J Immunol 139:55, 1987). This finding prompted us to investigate whether AIDS patients' effector cells could mediate ADCC against circulating CD4+ T cells infected with or expressing HIV antigen. The findings reported herein demonstrate that AIDS effector cells can mediate lysis of CEM (CD4+ T-cell line) coated with HIV protein in the presence of HIV-specific antibody. Lysis was specific, as non HIV-coated CEM or the addition of HIV-negative serum resulted in no lysis. We then examined HIV-coated peripheral blood-derived CD4+ T lymphocytes as targets in ADCC. We demonstrate that in the presence of HIV-specific antibody, HIV-coated CD4+ T lymphocytes serve as targets for ADCC by AIDS effector cells. The lytic activity obtained with AIDS effector cells was comparable to that obtained with normal effector cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975669 TI - Induction of interleukin-4-dependent IgE synthesis and interleukin-5-dependent eosinophil differentiation by supernatants of a human helper T-cell clone. AB - The human alloreactive T-cell clone A1 produces interleukin (IL)-4, IL-5, and granulocyte macrophage-colony stimulating factor (GM-CSF), but not IL-2 or interferon-gamma (IFN-gamma), as assessed by probing for mRNA transcripts, immunoassays, or bioassays. Supernatants from clone A1 induced IgE synthesis by normal human peripheral blood mononuclear cells. IL-4 was shown to be necessary and sufficient for the induction of IgE by clone A1 supernatants. In contrast, IgE induction by clone A1 supernatants and recombinant (r) IL-4 was inhibited by IFN-gamma. This suggests that the absence of IFN-gamma from the IL-4-containing A1 supernatants was important for their IgE-inducing ability. Supernatants from clone A1 could also specifically induce the growth of cord blood cell progenitors and their differentiation into eosinophils but not into basophils. rIL-5, but not rIL-4 or rGM-CSF, also induced eosinophil maturation. These data suggest that IL 5 secreted by clone A1 was responsible for its ability to induce eosinophil differentiation. The implications of the concomitant production of IL-4 and IL-5 by a single T-cell clone are discussed. PMID- 2975671 TI - Class II (DR) antigen expression on CD8+ lymphocyte subsets in acquired immune deficiency syndrome (AIDS). AB - In a selected group of human immunodeficiency virus (HIV)-infected patients we confirm the expansion of a CD8+ T-lymphocyte subset, i.e., the CD8+/Leu7+ cells, which account for 30% of the lymphocytes, compared to 3% in the control donors. In addition, a CD8+ T-lymphocyte subset that coexpresses class II (DR) antigens, i.e., CD8+/DR+ cells, is also increased from 1.5% in controls to 27% in the HIV infected patients. Using three-color immunofluorescence and flow cytometry we can demonstrate that the CD8+/Leu7+ and the CD8+/class II+ cells are not distinct but overlapping subsets. In the HIV-infected patients 42% of the CD8+/Leu7+ cells were strongly positive for class II and these CD8+/Leu7+/class II+ cells accounted for 13% of all lymphocytes. These findings indicate that the expanded CD8+/Leu7+ cells are activated and hence might be actively involved in immune defense in acquired immune deficiency syndrome (AIDS). PMID- 2975672 TI - Abnormal differentiation of immunoregulatory T-lymphocyte subpopulations in the major histocompatibility complex (MHC) class II antigen deficiency syndrome. AB - The major histocompatibility complex (MHC) class II deficiency syndrome is a rare immunodeficiency disease associated with defective expression of class II MHC antigens. We have examined the consequences of this defect for the differentiation and functional capabilities of immunoregulatory T-cell subpopulations in an affected patient. Although the number of circulating T cells was normal, there was a striking reduction in the number of CD4+ T cells. Furthermore, purified CD4+ cells from the patient were unable to provide help for antibody secretion. This defect in helper function appeared to be due to the abnormal differentiation of the few CD4+ cells present, virtually all of which expressed the CD4+HB11+ phenotype characteristic of immature "virgin" T cells. Abnormal development of immunoregulatory CD8+ T cells was also observed. Although increased numbers of CD8+ T cells were present, virtually none had phenotypic properties of suppressor cells (i.e., CD3+/CD8+/9.3- granular lymphocytes that coexpress the Leu-15 or Leu-7 antigens), and purified CD8+ cells from the patient had no suppressor activity. Thus, the absence of class II MHC antigens profoundly disrupts the development of immunoregulatory T cells. We propose that these effects occur by the following mechanisms: (1) the absence of intrathymic class II antigens results in deficient production of CD4+ cells, (2) the CD4+ cells that do emerge from the thymus do not undergo postthymic maturation into CD4+HB11 cells with helper capabilities, and (3) the absence of CD4+HB11- effector cells results in abortive development of suppressor cells involved in feedback suppression. PMID- 2975673 TI - [Studies of beta-endorphin and methionine-enkephalin in cerebral vascular disease. Its evaluation and clinical significance of periodic changes]. PMID- 2975675 TI - Localization of immunoreactive GABA and enkephalin and NADPH-diaphorase-positive neurons in fetal striatal grafts in the quinolinic-acid-lesioned rat neostriatum. AB - Fetal striatal tissue grafts have been shown to partially reverse the biochemical and behavioral deficits induced by excitotoxic lesions. To determine if grafted striatal neurons contain neurochemical markers similar to those in neurons in the caudate nucleus and to establish the morphological characteristics and relative frequency of labeled neurons in the grafts, the localization of immunoreactive GABA and leucine-enkephalin (ENK) and of NADPH-diaphorase (NADPH-d) activity was examined in fetal striatal grafts at the light and electron microscopic levels. Striatal tissue from 17-day fetuses was grafted into the caudate nucleus of adult rats 1 week after intracaudate injections of either a low or high dose of quinolinic acid. At the light microscopic level, immunoreactive GABA and ENK and NADPH-d-positive neurons, processes, and punctate structures were present within adjacent sections of the same grafts. The frequency and morphological features of these labeled cell populations were similar in grafts placed into either minimally or extensively lesioned striata. Immunoreactive GABA and ENK neurons in the grafts constituted 28% and 13.5%, respectively, of the neuronal population of the graft and their mean diameters were 22 and 14% larger, respectively, than neostriatal neurons that contained the same chemical markers. NADPH-d-positive neurons in the grafts formed 3.5% of total grafted neurons and exhibited characteristics of neostriatal NADPH-d-containing aspiny cells, including medium sized somata, indented nuclei, and varicose dendrites. At the electron microscopic level most GABA-positive neurons in the grafts contained indented nuclei and most immunoreactive ENK somata had unindented nuclei. Dendrites and dendritic spines with GABA or ENK immunoreactivity were present in the grafts where they were postsynaptic to unlabeled axons. Immunoreactive GABA and ENK axon terminals formed synapses with unlabeled neuronal profiles in the grafts. These findings demonstrate that fetal striatal grafts contain chemically defined neuronal populations that form synaptic connections within the graft and share some features with corresponding cell groups in the neostriatum. These results provide an anatomical basis for the graft-induced recovery from behavioral and biochemical deficits caused by instrastriatal lesions reported in other studies. PMID- 2975674 TI - Brush border myosin filament assembly and interaction with actin investigated with monoclonal antibodies. AB - Monoclonal antibodies binding to epitopes in the rod portion of brush border myosin were used to study the mechanism of filament assembly and its role in myosin interaction with actin. The antibodies and their Fab fragments had specific effects on the size of the filaments assembled in vitro. Two antibodies (BM3 and BM4), directed against the tip of the myosin tail, completely inhibited myosin filament assembly. The other antibodies (BM1, BM2 and BM5), binding to other sites along the myosin rod, only partially blocked filament growth, and short filaments could be assembled. Thiophosphorylated brush border myosin filaments appeared slightly more stable to the effects of the antibodies than those composed of dephosphorylated myosin. Only one (BM3) of the antibodies which completely inhibited the assembly of new filaments was capable of disassembling preformed myosin filaments. The other antibody, BM4, partially disassembled filaments, leaving approximately 0.2-microns long 'cores', suggesting that polymerization in this myosin occurs by a biphasic mechanism, i.e. the formation of a stable nucleus of antiparallely packed molecules, followed by elongation. The antibodies BM1 and BM2 bound to myosin filaments generating a regular transverse pattern with a approximately 14-nm periodicity, and had little effect on the stability of these preformed filaments. Inhibition of filament formation and solubilization of the myosin by the antibodies appeared to be associated with inhibition of myosin interaction with actin, as measured by the actin-activated MgATPase activity. In the presence of the antibodies which completely inhibit filament assembly, we observed a decrease to approximately 20% (BM4-Fab) and to approximately 50% (BM3) of the control actin-activated myosin MgATPase activity, and this activity was kinetically different from that of the soluble myosin S1 fragment, suggesting that the rod has a profound effect on the kinetics of actomyosin interaction. PMID- 2975676 TI - Secondary cutaneous cryptococcosis: case report and review of 22 cases. AB - A 51-year-old immunosuppressed man presented with ulcerations of the abdominal wall. Biopsy of the ulcer margin was necessary for the diagnosis of cutaneous cryptococcosis. Since a variety of opportunistic organisms can present with non specific cutaneous lesions, these infections must be ruled out by tissue diagnosis of all immunosuppressed patients. A review of 22 additional cases is included together with electron microscopic findings. PMID- 2975677 TI - Precautions reported to be used against cross-infection and attitudes to the dental treatment of HIV-positive patients in routine clinical dental practice in Scotland. PMID- 2975678 TI - Lichenoid eruption due to mercaptopropionylglycine. PMID- 2975679 TI - Social functioning in community residents with depression and other psychiatric disorders: results of the New Haven Epidemiologic Catchment Area Study. AB - Social functioning was compared among 4913 community participants with current depression, past depression, other psychiatric disorders and no psychiatric history, from the New Haven Epidemiologic Catchment Area Wave I survey. Respondents with current major depressive disorder (1.5%) (based on the Diagnostic Interview Schedule) reported significantly poorer intimate relationships and less satisfying social interactions than respondents with past depression or other current disorders. Respondents with no psychiatric history (77%) reported significantly more active and satisfying social interactions than persons with any psychiatric disorder. These social functioning and depression associations were similar among males and females, and corroborated results from patient samples. PMID- 2975680 TI - Sleep EEG recordings in generalized anxiety disorder with significant depression. AB - After one accommodation night, sleep EEG recordings were performed during three consecutive nights in ten drug-free inpatients presenting generalized anxiety disorder (GAD) with significant depression, compared with a age- and sex-matched group of patients with GAD and a group of primary major depressive disorder (MDD) patients. GAD patients with depression did not differ from GAD patients in any sleep variable. Patients with MDD showed more stage shifts and a greater number of awakenings than patients with GAD. REM latency was significantly shorter in MDD patients than in the other groups, and may thus help to differentiate anxious from depressed patients. PMID- 2975681 TI - Sex-related differences in the relationships between self-rated depression and biological markers. AB - Gender-related differences in self-reported depression, in biological factors putatively related to depression and in the associations between severity of illness and biological factors were investigated. To this end the Zung Self Rating Depression Scale (SDS), the ratio L-tryptophan/valine + leucine (L TRP/CAA) and basal cortisol in serum at 8 a.m. were determined in 51 depressed inpatients undergoing a dexamethasone suppression test (DST). In the total study group no significant relationships were established between severity of illness and either of the biological markers. In women, SDS correlated significantly (P less than 0.01) negatively with the ratio L-TRP/CAA and positively with post dexamethasone cortisol (P less than 0.01). In men these relationships tended to be inverted. The differences in the two sexes between these correlation coefficients were significant (P less than 0.01). These gender-related differences in the relationships between self-reported depression and the biological variables could be explained by differential psychoneuroendocrine and psychobiochemical responses. Future work on the severity of illnesses in terms of biological factors must take into account these differential responses between depressed males and females. PMID- 2975682 TI - A kinetic analysis of platelet monoamine oxidase activity in patients with panic attacks. AB - A kinetic analysis of platelet monoamine oxidase (MAO) was performed in 22 patients with panic disorder and 14 normal controls, using tyramine as a substrate. Patients and controls did not differ significantly for either Km or Vmax, when same sex contrasts were carried out. The results do not support some studies that have shown elevated MAO activity in patients with panic attacks. The reasons for the discrepancies between previous studies and the present one are explored. PMID- 2975683 TI - Cortisol measures in primary major depressive disorder with hypersomnia or appetite increase. AB - Morning plasma cortisol response to the 1 mg dexamethasone suppression test along with cortisol levels in blood, cerebrospinal fluid (CSF), and urine were measured in hospitalized male and female patients with primary major depressive disorder who reported hypersomnia (n = 23), or increase in appetite (n = 22). Comparisons were drawn to cortisol levels in patients with primary major depressive disorder who did not report hypersomnia or appetite increase (n = 23) and to normal controls (n = 23), all age- and sex-matched. Depressives with hypersomnia or increased appetite showed higher than normal 24-h urinary free cortisol concentrations. Depressed patients without hypersomnia or appetite increase had in addition to elevated free urinary cortisol concentrations higher than normal morning plasma cortisol levels before and after dexamethasone administration and a higher incidence of cortisol non-suppression after dexamethasone compared to normal subjects. The findings provide preliminary evidence that HPA activation in depression is diminished in the presence of hypersomnia and/or an increased appetite. Studies of the hypothalamic-pituitary-adrenal axis may be useful for differentiating subtypes of depression characterized by hypersomnia or enhanced appetite. PMID- 2975684 TI - Atenolol treatment of late luteal phase dysphoric disorder. AB - Activation of the renin-angiotensin-aldosterone system has been hypothesized as a potential pathophysiological factor in premenstrual tension syndrome (PMS). Atenolol is a predominate beta 1-blocker which can decrease plasma renin activity and inhibit the urinary excretion of aldosterone. Sixteen women meeting provisional diagnoses of late luteal phase dysphoric disorder were treated for symptoms of PMS with atenolol (50 mg once daily) in a randomized placebo crossover double-blind design. The data indicated significant improvements on the irritability, vigor, elation, and friendliness scores in response to atenolol compared to placebo. Significant changes were not found for several other ratings scales, indicating that atenolol improved only selected symptoms in the group as a whole. However, the women who had premenstrual tension symptoms for more than 5 years (n = 8) were improved on most of the rating scales. Atenolol decreased premenstrual plasma aldosterone to a limited extent. There was also a trend in the data toward higher luteal progesterone levels during the month subjects took atenolol. Plasma renin activity and aldosterone correlated with estrogen and progesterone levels during the placebo month but not during the active month. PMID- 2975685 TI - Parental representations of depressed outpatients from a U.S.A. sample. AB - A case-control study of adult depressed outpatients derived from a U.S. sample confirmed previous Australian reports of a significantly greater likelihood for depressives to report an earlier lack of parental care as well as parental overprotection. Only 32% of the patients, as against 62% of the non-psychiatric controls, reported 'optimal bonding', as defined by the Parental Bonding Instrument (PBI), from one or both parents. By contrast 62% of the depressives and 27% of the controls reported exposure to PBI-defined 'affectionless control' from one or both parents. As in previous studies, a raw PBI care score of less than 10 was highly discriminating, being reported by 32% of the depressives and only 3% of the controls. PMID- 2975686 TI - Study on suicide in depressed inpatients. AB - Seventy-five depressed clinical suicides were compared with 50 depressed non suicide clinical controls, using the method of stepwise logistic regression analysis. A set of the six best discriminating variables was identified, which comprised male sex, suicidal behavior at index admission and during index hospitalization, number of previous psychiatric hospitalizations, broken home and social exits. These factors should be taken into consideration when estimating suicidal danger in all types of depressed inpatients. PMID- 2975687 TI - Geographical variation in the seasonality of suicide. AB - This paper reports the results of a study on the seasonal variation of suicides occurred in Italy during the years 1969-1981. We found evidence for a cyclical pattern with one cycle per year for North/Central as well as for South Italy with no difference between the two parts in the variance explained by it. However, urban regions showed a higher seasonality than the rural parts of Italy, independently of their geographical location. To account for these findings, the hypothesis of a strong seasonal related urban rhythm has been put forward. PMID- 2975688 TI - Menstrual fluctuation in the symptoms of panic anxiety. AB - Ten women with DSM-III-defined panic attacks (five with and five without agoraphobia) had symptom severity rated daily, weekly, and retrospectively through one full menstrual cycle. Substantial fluctuations in retrospective ratings of severity were observed, with the premenstrual week being rated as most severe. Daily and weekly ratings showed much smaller fluctuations in the predicted direction. Possible reasons for this outcome are considered. PMID- 2975690 TI - Erythrocyte membrane proteins in psychiatric disorders and controls. AB - There are indications of abnormalities in membrane transport processes of erythrocytes in affective disorders. Therefore, it seemed worth while to investigate possible charge heterogeneities of erythrocyte membrane proteins in psychiatric disorders applying isoelectric focusing (IEF) in one- and two dimensional procedures. At this level of analytical sensitivity the protein patterns did not discriminate between major affective disorder, schizophrenia and controls. PMID- 2975689 TI - Are there distinct biochemical subtypes of depression? EEG characteristics of clinically defined on-drug responders and non-responders. AB - Subgroups of clinically defined responders and non-responders to clomipramine or maprotiline showed different electroencephalographic dynamics of vigilance, even before pharmacotherapy was initiated. Two hours after the first drug administration subgroup differences no longer existed. On the 21st day of medication we found a tendency towards re-establishing the pre-drug situation. Our findings lend support to the assumption of 'distinct biochemical subtypes' which, however, rather correspond to different stages of the pathophysiological process than represent invariant 'traits'. PMID- 2975691 TI - Seasonal variation of cortisol plasma levels in depressives. AB - There is some evidence for a seasonal variation of plasma cortisol levels after administration of dexamethasone in depressed subjects. This variation is suspected to be associated with similar season-related changes in pre dexamethasone cortisol levels. Therefore, cortisol plasma levels in depressed subjects before and after dexamethasone administration were investigated. No seasonal variation was found in pre- or post-dexamethasone cortisol levels. PMID- 2975692 TI - Inhibition of gonadotropic and ovarian function by intranasal administration of D Ser (TBU)6-EA10-LHRH in normo-ovulatory women and patients with polycystic ovary disease. AB - We investigated the effectiveness of D-Ser (TBU)6-EA10-LHRH (Buserelin) intranasally 600 micrograms/day given 6 times daily in desensitizing normal ovulatory women and patients with polycystic ovarian disease (PCOD) before initiation of ovarian stimulation for in vitro fertilization. We found that this regimen was sufficient to suppress the gonadotrophs in the normal women and in 8 out of 10 PCOD patients. In PCOD ovarian hormones became normal after Buserelin administration. Adrenal steroidogenesis was not affected by the GnRH agonist. We suggested that the frequency of administration of Buserelin was important to achieve a constant receptor binding and consequently a rapid desensitization. The choice of a monoclonal immunoradiometric assay for luteinizing hormone (LH) and follicle stimulating hormone (FSH) in association with the estradiol-benzoate provocation test were essential in evaluating desensitization. PMID- 2975694 TI - Renin-angiotensin and vasopressin in the development of salt-induced hypertension. AB - The purpose of this study was to assess the participation and interaction of the renin-angiotensin system and vasopressin in the early stages of the development of salt-induced hypertension. Subtotally nephrectomized rats, fed 1% saline, were treated over a 10-day period with either an antivasopressor V1 antagonist (by osmotic minipump) or an angiotensin converting enzyme inhibitor (either captopril or ramipril, given daily by oral gavage), or a combination of the two modes of treatment. Surprisingly, only ramipril (either alone or in combination with the V1 antagonist) could prevent the development of hypertension in these animals. Our data would not permit conclusion as to whether the different capacity of these agents to prevent salt-induced hypertension was due to a different degree of penetration into the central nervous system, or to some other property. PMID- 2975693 TI - Hormonal profile in benign breast disease. Endocrine status of cyclical mastalgia patients. AB - Plasma levels of prolactin, before and after TRH stimulation, progesterone, estradiol, sex-hormone-binding-globulin, androstenedione, testosterone and dehydroepiandrosterone-sulphate were determined in a group of women with benign breast disease and in control group, in order to correlate a possible hormonal disorder to breast pathology. The hormonal pattern in the patients with benign breast disease was characterized by an increased responsiveness of lactotrophes to TRH stimulation and by higher levels of sex hormone binding globulin, probably sustained by an underlying hyperestrogenism. PMID- 2975695 TI - Thalidomide and exfoliative dermatitis. PMID- 2975696 TI - Arthus-like phenomenon and lepromin A--a case report. PMID- 2975697 TI - Plasma atrial natriuretic peptide (ANP) in primary aldosteronism. PMID- 2975698 TI - Effects of percutaneous transluminal coronary angioplasty on left ventricular function evaluated by radionuclide angiocardiography. PMID- 2975699 TI - D1 and D2-type dopamine receptors in patients with Parkinson's disease and progressive supranuclear palsy. AB - The densities of D1- and D2-type dopamine receptors were measured with [3H]SCH23390 and [3H]spiperone, in the caudate nucleus and putamen of a large series of patients with Parkinson's disease or progressive supranuclear palsy, in relation to markers of dopaminergic and cholinergic innervation of the striatum ([3H]dihydrotetrabenazine binding and choline acetyltransferase activity). Correlations were sought between these parameters and clinical characteristics of the patients (abnormal involuntary movements, dementia, confusional syndrome or treatment). In Parkinson's disease, the densities of both types of receptors were unchanged, whereas in PSP, the density of D2, but not D1-type dopamine receptors, was decreased in the caudate nucleus and the putamen. No correlations between the biochemical and clinical data were found. PMID- 2975700 TI - Oculomotor defects in patients with Huntington's disease and their offspring. AB - We recorded saccadic, pursuit and fixation eye movements in patients (n = 5) with moderately advanced Huntington's disease (HD), offspring of HD patients (n = 22) and control subjects (n = 15), using the scleral sensor coil technique. Saccadic slowing was seen in all patients, no controls and (marginally) in a few at-risk subjects. Fixational stability was lower in patients than in the other groups; a structured background enhanced the difference and revealed decreased stability in a number of at-risk subjects. Smooth pursuit showed large errors in most patients and several controls but none of the at-risk subjects. Sporadic follow-up data show that at least two of the at-risk subjects developed manifest HD within a few years after passing the oculomotor test with entirely normal results. The material as a whole suggests that oculomotor dysfunction does not develop prior to, but concurrently with and as a part of generalized, progressive deterioration of motor control. The implication is that oculomotor screening of clinically healthy at-risk subjects does not reliably contribute to an earlier diagnosis of future HD. PMID- 2975701 TI - [The relation of antigen levels in the middle ear and round window membrane permeability in antigen-induced otitis media]. PMID- 2975702 TI - [Differentiation of hypertrophic cardiomyopathy from left ventricular hypertrophy induced by essential hypertension using magnetic resonance imaging]. AB - To examine the efficacy of magnetic resonance imaging (MRI) in diagnosing hypertrophic cardiomyopathy (HCM), 16 patients with HCM and 14 hypertensives with left ventricular hypertrophy (LDH) were studied using a 0.5 Tesla Siemens MRI apparatus equipped with cardiac gating. In HCM, left ventricular hypertrophy was localized to the septal wall in four, to the apical wall in two, to both the septal and apical walls in two, and to the apical and inferior walls in one, and it was diffuse in seven patients. In hypertensives, LVH was localized to the septal wall in three, to both the septal and anterior walls in two, to the free wall in one, and it was diffuse in eight patients. The distribution of the hypertrophic portion was nearly equal in both groups. The thickest portion of the left ventricular wall was 24.6 +- 4.8 mm in HCM and 21.6 +- 5.4 mm in hypertension, and there was no significant difference between them. The T2 relaxation time of the hypertrophic portion was 52.2 +- 4.8 msec in HCM and 45.3 +- 6.1 msec in hypertension, and there was a significant difference between them (p less than 0.01). However, there were no significant differences between the T2 relaxation times of the hypertrophic and non-hypertrophic portions in both groups. In conclusion, it may be difficult to differentiate HCM from hypertension based on the distribution of hypertrophic portions, but measurements of the T2 relaxation times may be useful for making the differential diagnosis. PMID- 2975703 TI - [Myocardial involvement in female Fabry's disease: evaluation by thallium-201 myocardial scintigraphy]. AB - Fabry's disease is characterized by an inherited X-linked disorder of glycosphingolipid catabolism, and heterozygous women affected with this disease who show overt symptoms including cardiac manifestations have rarely been reported. To elucidate the features of myocardial involvement in female patients, noninvasive techniques including exercise stress thallium-201 myocardial scintigraphy were performed. Three female patients, Cases 1-3, 26, 29 and 50 years of age, were documented low leucocytic alpha-galactosidase activities of less than 48% of normal (67.92-16.2 nmol/mg protein/h). They were examined using ECG, two-dimensional echocardiography (2-D Echo), Holter ECG, treadmill test and stress scintigraphy. On the ECG, negative T waves were shown in leads III and aVF in Cases 1 and 2. Left ventricular high voltage, giant negative T waves and short PR intervals were seen in Case 3. The 2-D Echo revealed neither valvular change nor left ventricular hypertrophy. On the Holter ECG, monofocal ventricular premature beats were occasionally observed in Cases 1 and 3. The treadmill test showed positive ST changes only in Case 2. On the exercise stress scintigraphy, uptake of thallium-201 was enhanced in the apex of the heart in Cases 2 and 3. Low uptake areas of thallium-201 were observed in Case 3. The ventricular angiogram revealed slight hypertrophy of the wall of the apical portion. In endocardial biopsies from the right ventricle, myelinoid lamellar inclusions were demonstrated in myocardial cells electron microscopically. Increased uptake of thallium-201 in the apex was noted in two of the three patients, but no apical thickening was noticed in any of the three cases by 2-D Echo. From the result of the biopsy of Case 3, the increased apical uptake of thallium-201 seems to reflect thickening caused by the deposition of glycosphingolipid. It was concluded that myocardial involvement in female Fabry's disease may occur early in the third decade and that the lesions could be detected with high sensitivity by thallium-201 myocardial scintigraphy. PMID- 2975705 TI - Old age in Down's syndrome. PMID- 2975704 TI - [Indications for coronary revascularization and the postoperative evaluations using Tl-201 exercise myocardial scintigraphy and a bull's eye display]. AB - Tl-201 exercise myocardial scintigraphy and quantitative analysis using bull's eye display were performed in 31 cases (18 bypass cases, 13 PTCA cases) to determine the indications for coronary artery bypass surgery and PTCA, and to evaluate postoperative improvement. Regions of interest (ROI) corresponding to each revascularized area were created on a bull's eye display. Then the washout rates (WR) and % uptakes were expressed as percentages. Improvement was judged to have occurred if the revascularized coronary artery was patent and both the WR and % uptake returned to the normal range as determined from the examinations of 20 normal cases. The results were as follows: 1. The preoperative mean WR of the improved areas (30 vessels) was 19 +- 15%, while that of the unimproved areas (15 vessels) was 35 +- 7%. We assumed that the area was suitable for revascularization when the preoperative WR was less than 25% which was in the lower limit of the normal range. Then, 18 vessels were judged to be suitable for surgery, and subsequent postoperative improvement was obtained in the 17 areas. 2. In 32 scintigraphically-improved areas, 30 vessels were angiographically patent, while five vessels were obliterated angiographically in 13 scintigraphically-unimproved areas (diagnostic validity was 76% of all 45 vessels). 3. The exercise tolerance increased significantly (p less than 0.01) from 8.7 +- 2.0 min to 11.6 +- 2.5 min in the improved cases in which all the revascularized areas were improved after revascularization. There was no change of the exercise tolerance (before: 9.4 +- 3.4 min, after: 9.4 +- 2.5 min) in the unimproved cases in which all the revascularized areas were unimproved. 4. Quantitative analysis was useful for objective evaluation, because the visual evaluations did not always agree with the quantitative evaluation. We concluded that the area with the preoperative WR less than the normal range is suitable for revascularization. As the scintigraphic evaluation was in accord with improved exercise tolerance and with patency as observed by coronary angiography, our method seems useful for postoperative follow-up. PMID- 2975706 TI - Exaggerated atrial natriuretic peptide release during acute exercise in essential hypertension. AB - The effects of acute exercise on plasma concentrations of atrial natriuretic peptide (ANP), arginine vasopressin (AVP), and plasma renin activity (PRA) were studied in 13 patients with previously untreated essential hypertension, and 8 matched normotensive control subjects. Resting levels of ANP and PRA were similar in the two groups, while resting AVP concentrations were 1.4 times higher in hypertensive subjects. Graded exercise was performed on a bicycle ergometer with workload increased each minute until exhaustion (Wmax). Wmax was higher in normal subjects than in hypertensive patients. Blood pressure and heart rate rose more steeply in hypertensive patients. Plasma ANP increased during acute exercise in both groups, but the average increase in hypertensives was substantially greater than in normal subjects (P less than 0.05). The increase in ANP during exercise was greater in hypertensives with left ventricular (LV) hypertrophy, and there was a positive correlation between LV mass and the percentage rise in ANP during exercise (r = 0.56, P less than 0.005). Plasma AVP did not alter during exercise. Plasma renin concentrations showed a small rise during exercise in both groups, which was 16% less in hypertensive subjects (P less than 0.05). The enhancement of ANP release during exercise in hypertensive subjects may reflect both cardiac structural changes and increased redistribution of blood to the cardiopulmonary compartment. PMID- 2975707 TI - Three-dimensional model of stellacyanin and its implications for electron transfer reactivity. AB - Experimental data were combined with computational methods in constructing a hypothetical three-dimensional model for the blue single copper protein Rhus stellacyanin (St). The known sequence of stellacyanin and its homology with plastocyanin (Pc) were used together with the results of spectroscopic studies of the protein that yielded the current assignment of two histidines, one cysteine and a disulfide sulfur as copper ligands in stellacyanin. By computer graphics and energy minimization the folding of the protein was predicted. The model structure is somewhat less regular than Pc as judged by surface area and energy comparisons, but it is a stable structure. Besides rotation of one imidazole ring the copper site undergoes no change even in the absence of the copper ion and the model shows that the site can be constructed with the four assumed copper ligands without forming a strained system. The structure also indicates that a carbonyl oxygen atom is near the copper, thus the site may have analogy to the Alcaligenes denitrificans azurin (Az) site, although the amino acid sequence is more homologous to that of Pc. The model indicates that aspartate 49, reductively labeled by Cr(III), is near the copper center and homologous to the site labeled by Cr(III) on Pc. Also homologous to Pc is a tyrosine residue adjacent to the aspartate. This tyrosine has been implicated in Pc electron transfer and thus is probably involved in electron transfer reactivity of St as well. The higher reactivity of St with small-molecule redox reagents compared to Az and Pc, may be due to the proximity of the above-mentioned aspartate 49 to the Cu, or the greater exposure of one of the Cu cysteine ligands, in the predicted structure as compared to that in the known Pc and Az structures. PMID- 2975708 TI - Sequence analysis of mitochondrial DNA from Podospora anserina. Pervasiveness of a class I intron in three separate genes. AB - A 48 kb region of the 95 kb mitochondrial genome of Podospora anserina has been mapped and sequenced (1 kb = 10(3) base-pairs). The DNA sequence of the genes for ND2, 3, 4, ATPase 6 and URFC are presented here. As in Neurospora crassa, the ND2 and 3 genes consist of a unit separated by one TAA stop codon. ND3, 4 and ATPase 6 are interrupted by class I introns. All three introns are remarkably similar in the C-domain of their secondary structure, sufficient enough to designate them as new subgroup, class IC introns. The open reading frames of the ND3 and 4 introns bear a high sequence similarity to the open reading frame of the class IB introns of ATPase 6 from N. crassa and ND1 from Neurospora intermedia Varkud. We also show that the tRNA Met-2 gene is duplicated and is involved in a recombinational event. The 5' region of URFC is also duplicated but no involvement of this gene with recombination or formation of plasmids is known. The evolutionary significance of the similarities of intron secondary structures and open reading frames of the ND3, 4 and ATPase 6 genes is discussed, including the possible separate evolution of structural and coding sequences. PMID- 2975709 TI - Crystal structure of the complex of phosphofructokinase from Escherichia coli with its reaction products. AB - The crystal structure of Escherichia coli phosphofructokinase complexed with its reaction products fructose 1,6-bisphosphate (Fru1,6P) and ADP/Mg2+, and the allosteric activator ADP/Mg2+, has been determined at 2.4 A resolution. The structure was solved by molecular replacement using the known structure of Bacillus stearothermophilus phosphofructokinase, and has been refined to a crystallographic R-factor of 0.165 for all data. The crystallization mixture contained the substrate fructose 6-phosphate, but the electron density maps showed clearly the presence of the product fructose 1,6-bisphosphate, presumably formed by the enzyme reaction with contaminating ATP. The crystal consists of tetrameric molecules with subunits in two different conformations despite their chemical identity. The magnesium ion in the "closed" subunit bridges the phosphate groups of the two products. In the "open" subunit, the products are about 1.5 A further apart, with the Mg2+ bound only to ADP. These two conformations probably represent two successive stages along the reaction pathway, in which the closure of the subunit is required to bring the substrates sufficiently close to react. This conformational change within the subunit is distinct from the quaternary structure change seen previously in the inactive T state conformation. It is probably not involved in the co-operativity or allosteric control of the enzyme, since the co-operative product fructose 1,6 bisphosphate is not moved, nor are the subunit interfaces changed. The structure of the enzyme is similar to that of B. stearothermophilus phosphofructokinase, and confirms the location of the sites for the two reaction products (or substrates), and of the effector site binding the activator ADP/Mg2+. However, this structure gives a clearer picture of the active site, and of the interactions between the enzyme and its reaction products. PMID- 2975710 TI - The effects of triiodothyronine on cultured neonatal rat cardiac myocytes. AB - The purpose of this study was to determine whether thyroid hormone could directly affect the phenotypic expression of two isozymic systems [lactate dehydrogenase (LDH) and myosin] and the energy transducing potential of cultured neonatal heart cells. In addition we determined if these biochemical systems developed in culture as they normally do during in vivo post-natal development. Cells were maintained for 14 days in culture medium containing 10% horse serum and Earle's salts. Experimental cultures were supplemented with 10 nmol/l 3,3',5-triiodo-L thyronine (T3). Hearts used to study in vivo development were excised from rats at the ages of 2 and 14 days post-natal to correspond with the time of isolating and harvesting the cultured heart cells, respectively. Adult hearts were used to represent the final developmental stage. Cultured cardiomyocytes without T3 administered to the culture medium showed no change in the isozymic profiles (myosin and LDH) or in metabolic potential during the 2 week culture period. The T3 treated cultures showed a complete shift to the V1 myosin isozyme. The glycolytic and aerobic metabolic potential [i.e., phosphofructokinase (PFK) and citrate synthase (CS) activities] and the LDH isozyme distribution were unaltered by T3 treatment. During in vivo development a shift toward the V1 myosin and H LDH isozymes along with an increase in aerobic metabolism occurred in the rat heart. These findings indicate that the development of these selected biochemical systems in cultured cardiac myocytes does not result from an intrinsic myogenetic program and thus must be regulated in vivo by epigenetic factor(s). These results show that T3 has the potential to be the prime determinant of the phenotypic expression of the myosin isoforms, but does not have the potential to be the sole determinant for the expression of the LDH isozymes or the glycolytic (PFK) and aerobic (CS) capacities of cardiac muscle cells. PMID- 2975711 TI - Regression of cardiac hypertrophy: morphometric and biochemical studies in rat heart after swimming training. AB - There is currently little information about the morphological changes of the myocardium accompanying the reversal of cardiac hypertrophy. In this study the hypothesis was tested that myocardial alterations induced by exercise will regress within a short interval after the end of training. Rat hearts were examined using morphometric and biochemical methods at the end of a 9-week period of endurance training and also 7, 10 and 14 days after its termination. At the end of the training period the heart weight had increased by 65% but the weight ratio of the right and left ventricular wall remained unchanged. A decline in the DNA content by 27% as well as a decrease in the volume density of the interstitial space by 14% and in the number of interstitial cell nuclei by 32% against controls, are explained by a 30% increase in the width of myofibres. The capillary density was reduced by 22% but the volume density of capillaries remained nearly constant as a result of widening of the capillary diameter by 27%. The surface density of capillaries was diminished by 10%. Ultrastructurally an increase in the ratio of mitochondrial to myofibrillar volume density was observed in the myocytes of hypertrophied hearts as compared to controls (0.54 and 0.63, respectively). Fourteen days after termination of training, 80% of the increment in heart weight had regressed. At this time the width of the myofibres and the volume density of the interstitial space had nearly normalized, while the capillary to fibre ratio had significantly increased. The ratio of mitochondrial and myofibrillar volume density became nearly normal, and a confluence of intermyofibrillar mitochondria resulted in significantly longer organelles. The increased DNA content 10 days after the training, as compared to controls, is attributable to the genesis of non-myocardial cells during the hypertrophic growth and their persistence during regression. The study has shown that cardiac hypertrophy induced by physical training nearly completely regresses within 14 days after termination of conditioning. The increased capillary to fibre ratio indicates neoformation of transversely oriented capillary branches in hypertrophy which particularly becomes apparent in two-dimensional estimation in the regression period. In comparison with myofibres, regression of capillaries seems to be delayed. The decline of heart weight and a significantly diminished RNA content during the regression of hypertrophy suggest that reduced synthesis is responsible for the decrease in heart weight. PMID- 2975712 TI - Acute and chronic effects of xamoterol in idiopathic dilated cardiomyopathy. AB - Xamoterol, a beta 1-partial agonist, was given to 10 patients with idiopathic dilated cardiomyopathy in NYHA functional classes II and III. The acute and chronic effects of xamoterol were assessed by changes in effort tolerance measured by multistage bicycle ergometry, echocardiography, radionuclide ventriculography and right heart catheterization. The acute effect was determined after a single intravenous injection of 0.2 mg/kg of xamoterol. Exercise heart rate decreased from 117 +/- 10 (mean +/- 1 SD) to 97 +/- 19 beats/min (p less than 0.01), but blood pressure and cardiac index were unaffected. When the drug was given orally at 200 mg/day for 3 months, exercise duration increased from 4.2 +/- 2.3 to 6.4 +/- 2.4 min (p less than 0.01), echocardiographic ventricular ejection fraction from 29 +/- 12 to 33 +/- 14% (NS) and radionuclide left ventricular ejection fraction from 37 +/- 11 to 45 +/- 14% (p less than 0.01), while pulmonary wedge pressure during exercise decreased from 40 +/- 10 to 31 +/- 10 mmHg (p less than 0.01). It may be concluded that xamoterol, by its action as a partial agonist, demonstrated both beta-agonist and antagonist effects in patients with idiopathic dilated cardiomyopathy. It thereby had beneficial effects on left ventricular function with resultant improvements in effort tolerance and subjective symptoms during long term treatment. PMID- 2975713 TI - The curvilinear relationship between aortic regurgitant volumes and color flow jets. Intraoperative two-dimensional Doppler and electromagnetic flowmeter correlation. AB - The potential usefulness of two-dimensional Doppler to quantitate valvular regurgitation depends on the relationship between the dimensions of the velocity maps and the regurgitant volumes. We previously found a significant linear regression between aortic regurgitant fraction and jet area, but because of a shallow slope, postulated an exponential or curvilinear relationship. A retrospective analysis of intraoperative electromagnetic flowmeter measurements of aortic flow and epicardial color flow maps of regurgitant jets gave us a more direct examination of the relationship over a wider range of lower flows. Regurgitant volume was calculated from reverse aortic flow and was compared to jet length, area (by planimetry) and volume (from Simpson's rule). Both linear and curvilinear regressions were determined. Correlation coefficients were similar but consistently closer to unity with the exponential equations, and data points were more closely distributed along a curved rather than a straight line. Length regressions were flatter and the volume regressions were steeper than the slopes of the area equations. Thus, the relationship of the gradual slope of the color flow jet area to the regurgitant volume is geometrically determined and limits quantitative application. PMID- 2975714 TI - [Brain pathology in Alzheimer's disease. Ultrastructure of amyloid fibrils in Alzheimer's disease and Down's syndrome]. PMID- 2975715 TI - [Cytogenetic study and chromosome abnormalities of Alzheimer's disease and Down's syndrome]. PMID- 2975716 TI - [Kidney cancer diagnosed by radionuclide angiography in a patient with heart disease]. PMID- 2975717 TI - [Induction of mucin deposition in mouse skin by serum from a patient with lichen myxedematosus]. PMID- 2975718 TI - Response of atrial natriuretic peptide in plasma and urine to changes in dietary intake of sodium chloride in man. AB - Plasma concentration and urinary excretion of immunoreactive human atrial natriuretic peptide (ir-hANP), aldosterone, and vasopressin were measured, and renal function and blood pressure were determined in six healthy male subjects in three periods of different sodium intake: the control (172 mEq/day), low (20 mEq/day), and high (285 mEq/day) sodium period. Both plasma concentration and urinary excretion of ir-hANP increased significantly in the high sodium period but did not change between the control and low sodium periods. On the contrary, aldosterone increased and vasopressin decreased in the low sodium period but did not change between the control and high sodium periods. These results may point out the reciprocal action of both endocrine systems. The glomerular filtration rate changed in parallel with sodium intake whereas the fractional excretion of sodium decreased only in the low sodium period, probably reflecting the action of aldosterone. It is concluded from these results that the atrial peptide may be an important component in the regulation of body fluid in the high sodium loading of physiological range but its action may be limited to the control of glomerular filtration rate. PMID- 2975719 TI - Acute exhaustive exercise changes the metabolic profiles in slow and fast muscles of rat. AB - The effects of acute endurance running on the metabolic profiles of rat skeletal muscle were studied. Male Wistar strain rats were continuously run on a treadmill for 1 h (speed, 35 m/min; grade, 0 degrees). Soleus (SOL) and extensor digitorum longus (EDL) were removed after 30-min running, and a 0, 1, 6, 24, 48, and 72 h post-exercise, and enzymes activity (CK, LDH, PFK, PK, SDH, and MDH) and substrates contents (glycogen and pyruvate) were measured biochemically. The time course of the enzyme activities showed two distinct patterns: CK, LDH, SDH, and MDH showed two peaks, at 0 and 24 h post-exercise, while PFK and PK showed one peak at 0 h post-exercise. The activities of glycolytic enzymes and CK in EDL and oxidative enzymes in SOL showed marked changes after exercise. The glycogen level was lowest at 0 h post-exercise in both muscles and recovered to resting level by 24 h post-exercise. Pyruvate increased with running and showed the highest value at 1 h post-exercise. Increased oxidative capacity of skeletal muscle in response to the acute endurance exercise dropped gradually to the resting level by 48 h post-exercise. An endurance exercise may induce a flexible adaptation on the oxidative capacity within skeletal muscle. We conclude that the respective time course of the enzyme activities must be considered when discussing metabolic changes that occur with acute endurance exercise. PMID- 2975720 TI - Histochemical properties of intrafusal fibers in the soleus muscle of the aged rat. AB - ATPase reaction profiles of intrafusal fibers in the muscle spindle of the soleus muscle of 135-week-old rats were examined. Nuclear bag1 fibers contained an acid- and alkaline-labile form of the enzyme or an acid-labile and alkaline-stabile form, nuclear bag2 fibers contained an acid- and alkaline-stabile form, and nuclear chain fibers contained an acid-labile and alkaline-stabile form. These results indicate that the enzyme histochemical heterogeneity of intrafusal fibers is well-preserved during ageing. PMID- 2975721 TI - Release of atrial natriuretic peptide from the isolated, blood-perfused right atrium of the dog. AB - Release of atrial natriuretic peptide (ANP) was investigated using the isolated right atrium (RA) cross-circulated with heparinized arterial blood of the donor dog. ANP concentration of the blood was measured by radioimmunoassay. The plasma ANP concentration of the arterial blood of the donor dog (ANP-D) which perfused the RA preparation was 111 +/- 18 pg/ml (n = 4), while the ANP concentration of venous blood leaving the RA preparation (ANP-A) was 1680 +/- 220 pg/ml (n = 4). In comparison, the plasma ANP concentration of venous blood leaving the isolated papillary muscle of the right ventricle (ANP-V) was 106 +/- 19 pg/ml (n = 4), which was not significantly different from the ANP-D concentration. When the right atrium was electrically driven at a rate of 100 beats/min, almost the same rate as the spontaneous sinoatrial rate of 98 +/- 6 beats/min (n = 4), the ANP-A concentration was not changed (1630 +/- 160 pg/ml, n = 4). When the driving rate was increased to 200 beats/min, the ANP-A concentration was significantly increased to 2250 +/- 130 pg/ml (n = 4). These results suggest that ANP is exclusively secreted from the atrium, but not from the ventricle, and that release of ANP is directly related to atrial rate. PMID- 2975723 TI - [Biochemical analysis of human external urethral sphincter]. PMID- 2975722 TI - Effect of streptozotocin administration to pregnant mice on serum glucose, glycosylated hemoglobin and weight of organs of the mother mice and their pups. AB - Administration of streptozotocin (STZ, 200 mg/kg, i.p.) to pregnant mice during either the early or middle stages of pregnancy produced diabetic conditions in the mother mice. It reduced the litter size, but did not induce diabetes in the pups. The weights of the liver and kidney per unit body weight of the infants of STZ-treated mother mice were slightly larger, though not significantly, than that of infants from control mothers; however, no increase was observed in the blood glucose or glycosylated hemoglobin (HbA1) values. PMID- 2975724 TI - [Effect of percutaneous intravascular coronaroplasty on left-ventricular diastole in patients with unstable angina]. PMID- 2975725 TI - [Echocardiogram and activity of the sympathetic nervous system in patients with primary arterial hypertension]. PMID- 2975726 TI - [Disorganization of myocardial fibers. I. Pathophysiological principles. Methods of study]. PMID- 2975727 TI - [Disorganization of myocardial fibers. Results of the studies. The role in the pathogenesis of cardiac hypertrophy]. PMID- 2975728 TI - [Beta-endorphin in the blood of patients with pheochromocytoma]. PMID- 2975730 TI - [Hospital ambulatory surgical services for children]. PMID- 2975729 TI - [Organization of the provision of emergency medical services on ocean-going ships]. PMID- 2975731 TI - [Means of improving the practical training of subordinate surgeons at a surgical clinic]. PMID- 2975732 TI - [Methodologic requirements for the independent work of students at a department of surgery]. PMID- 2975733 TI - [Optimization of the teaching of practical skills to students at a department of surgical diseases]. PMID- 2975734 TI - Rederivation of inbred strains of mice by means of embryo transfer. PMID- 2975735 TI - Murine encephalitozoonosis: the effect of age and mode of transmission on occurrence of infection. AB - Experiments were conducted to determine whether neonatal mice are more susceptible to E. cuniculi than adult mice, and whether vertical and/or horizontal transmission occur in murine encephalitozoonosis. E. cuniculi infection in neonates did not cause mortality or clinical signs, but did result in chronic infection. Despite initial age-related immunodeficiency, mice infected as neonates eventually developed humoral and cell-mediated immune responses against the parasite comparable to those seen in adult mice. The results suggested that neonatal mice are not more susceptible to E. cuniculi than adult mice. Pups from either infected or normal parents did not differ in humoral and cell-mediated immune responses after challenge, suggesting that pups from infected parents were not infected with E. cuniculi during gestation. In contrast, mice became infected by caging with infected mice demonstrating that horizontal infection does occur. PMID- 2975736 TI - Modulated immune responsiveness associated with experimental Encephalitozoon cuniculi infection in BALB/c mice. AB - Spleen cell blastogenesis to mitogens and antibody responses to sheep erythrocytes (sRBC) were tested in BALB/c mice with experimental E. cuniculi infections. Blastogenesis responses of spleen cells 1 week post-infection were significantly lower than normal to T-cell mitogens (Con A and PHA) and were unchanged in response to B-cell mitogens (LPS and PWM). After 2 weeks post infection, the responses to T cell mitogens returned to normal. Mixing spleen cells from 1-week infected mice with cells from uninfected mice failed to reveal the presence of suppressor cells. Antibody responses to sRBC were significantly slower to develop in 1 week-infected mice compared with uninfected mice or mice infected 2 weeks earlier or at the same time as sRBC challenge. Infected mice displayed splenomegaly which was most pronounced 1 week post-infection and the differential spleen cell counts revealed the presence of lymphoblasts. Lymphohyperplasia appeared to cause the splenomegaly. No shifts in the proportion of Thy 1.2+ T cells, Ig+ B cells, or esterase-positive macrophages were detected. These results indicate that the immune system in BALB/c mice is depressed early during E. cuniculi infections. PMID- 2975737 TI - A silver impregnation method for motor and sensory nerves and their endings in formalin-fixed mammalian muscles. AB - A silver impregnation method is described which shows motor and sensory nerves and their endings in formalin-fixed mammalian muscles. The method works with the same reliability on flattened muscle pieces as well as on frozen sections. Large nerve bundles, myelinated and non-myelinated single axons, and terminals impregnated by this method stand out black against a light brown background. PMID- 2975738 TI - Transluminal dilation techniques in the management of congenital obstructive cardiovascular disease. PMID- 2975739 TI - Influence of dimethylacetamide, N-N-diethyl-m-toluamide and 1 dodecylazacycloheptan-2-one on ex vivo permeation of phosphonoformic acid through rat skin. AB - Transdermal permeation of trisodium phosphonoformate (PFA) alone and in presence of sorption promoters, dimethylacetamide (DMAC), N-N-diethyl-m-toluamide (NNDEMT) and 1-dodecylazacycloheptan-2-one (Azone) was studied using excised rat skin. DMAC in concentrations between 0.05% to 10% had no effect on amount of PFA in the skin or permeated across skin, flux or lag time. 10% NNDEMT doubled the amount of PFA in the skin, increased fourfold the amount permeated across the skin, and increased the flux fivefold. There was no influence on lag time. Azone doubled the amount of PFA in the skin, more than tripled the amount permeated across the skin, and increased the flux fourfold. There was no influence on lag time. PMID- 2975742 TI - [Certification of work places]. PMID- 2975740 TI - [How to lay out a health bulletin]. PMID- 2975741 TI - [How to lay out a hospital]. PMID- 2975743 TI - [Certification of the work places of the duty nurses in the therapeutic institutions of Grodno Province]. PMID- 2975744 TI - [Raising the qualifications of paramedical and pharmaceutical personnel in the Russian Federation]. PMID- 2975745 TI - [The work experience of the head nurse of a province]. PMID- 2975746 TI - [Diagnostic usefulness of a monoclonal antibody against the common leukocyte antigen]. PMID- 2975747 TI - The caries inhibitory effects of GOS-sugar in vitro and in rat experiments. AB - The caries inhibitory activity of GOS-sugar (panose- and maltose-rich sugar mixture) was examined and compared with that of sucrose, maltose, or glucose in in vitro and in vivo experiments. Streptococcus mutans MT8148R (serotype c) and Streptococcus sobrinus 6715 (g) did ferment GOS-sugar and produce acid in a similar way as with maltose and glucose. However, GOS-sugar could not be a substrate for the glucosyltransferases (GTases) of these mutans streptococci to synthesize the water-insoluble glucan. Also, it significantly inhibited not only the synthesis of water-insoluble glucan from sucrose by the crude GTases but also the sucrose-dependent adherence of these cells to a glass surface. In particular, adherence of growing cells of 6715 was markedly inhibited by the presence of GOS sugar. GOS-sugar was found to induce significant but minimal dental caries in SPF rats infected with either MT8148R or 6715. Furthermore, the replacement of half of the dietary sucrose content with GOS-sugar resulted in a significant reduction of caries development in rats infected with strain 6715. PMID- 2975748 TI - [Effects of serum immunosuppressive factors on the cytotoxicity of lymphokine activated killer (LAK) cells induced by recombinant interleukin 2(R-IL2)]. AB - Fundamental studies were performed on adoptive immunotherapy, especially on effects on lymphocytic cytotoxic activity, of nonspecific immunosuppressive factors (ferritin, IAP, AFP) and of serum factors obtained from gastric cancer patients, and possible intervention of suppressor T cells in serum immunosuppressive activity on the cytotoxicity was also examined. The following results were obtained. 1) Cytotoxicity of LAK cells induced by culturing normal peripheral blood lymphocytes (PBL) with R-IL2 in the medium containing normal AB type sera, was higher than that of PBL. 2) Effect of nonspecific immunosuppressive factors on cytotoxicity of LAK cells was lower than that on cytotoxicity of PBL. 3) Cytotoxicity of PBL was inhibited in a relatively specific fashion by sera from patients of the cancers which were of identical histological types with the target tumor cells, while that of LAK cells was hardly inhibited by patients' sera. 4) Cytotoxicity of Leu 15-PBL was inhibited by nonspecific immunosuppressive factors and also by cancer patients' sera in a relatively specific fashion in relation to histology. 5) Cytotoxicity of Leu 15 LAK cells was hardly inhibited by serum nonspecific and specific immunosuppressive factors. The above results showed that serum immunosuppressive factors might act on PBL cytotoxicity without intervention of suppressor T cells, and that LAK cells were hardly inhibited by such immunosuppressive factors. All these results suggested usefulness of adoptive immunotherapy with LAK. PMID- 2975749 TI - Characterization of a mutation that causes overproduction of inositol in Neurospora crassa. AB - Slow-growing (inl+/-) spontaneous mutants have been isolated from an inositol requiring (inl) strain of Neurospora crassa that produces defective myo-inositol 1-phosphate synthase (MIPS), the enzyme responsible for the production of inositol-1-phosphate from glucose-6-phosphate. The defective enzyme has some residual activity. In the inl+/- strain the synthesis of the defective enzyme is enhanced, which enables the strain to grow slowly on minimal medium. The mutation (opi1) responsible for the partial inositol independence segregates independently from the inositol locus, and suppresses the inositolless character by overproduction of defective MIPS. opi1 acting upon the wild type (inl+) allele increases MIPS production and causes inositol excretion. PMID- 2975750 TI - Influence of exercise on cardiac and skeletal muscle myofibrillar proteins. AB - The purpose of this study was to examine the Ca2+-Mg2+ myofibrillar ATPase and protein composition of cardiac and skeletal muscle following strenuous activity to voluntary exhaustion. Sprague-Dawley rats (200 g) were assigned to a control and exercised group, with the run group completing 25 m.min-1 and 8% grade for 1 hour. Following activity, the myocardial Ca2+-Mg2+ myofibrillar ATPase activity pCa relationship had undergone a rightward shift in the curve. Electrophoretic analysis revealed a change in the pattern of cardiac myofibrillar protein bands, particularly in the 38-42 Kdalton region. Enzymatic analysis of myofibrillar proteins from plantaris muscle, revealed no change in Ca2+ regulation following exercise. Electronmicrographic and electrophoretic analysis revealed extensively disrupted sarcomeric structure and a change in the ratio of several plantaris myofibrillar proteins. No difference was observed for myosin: Actin: tropomyosin ratios; however a dramatic reduction in 58 and 95 Kdalton proteins were evident. The results indicate that prolonged running is associated with similar responses in cardiac and skeletal muscle myofibrillar protein compositions. The abnormalities in myofibrillar ultrastructure may implicate force transmission failure as a factor in exercised-induced muscle damage and/or fatigue. PMID- 2975751 TI - Comparison of ATPase activities and heavy chains of rabbit atrial and thyrotoxic ventricular myosin subfragment-1. AB - Subfragment-1 of rabbit atrial and thyrotoxic ventricular myosin (V1 isomyosin) has been prepared and purified by DEAE-cellulose column chromatography. Pyrophosphate-polyacrylamide gel electrophoretic patterns and column chromatographic profile of the atrial subfragment differ from those of thyrotoxic ventricular myosin subfragment-1. On the other hand, Ca2+, Mg2+ and actin activated ATPase activities of these subfragments are identical. Comparison of the peptide mapping by limited proteolysis in the presence of sodium dodecyl sulfate of the heavy and the light subunits of these subfragments reveals that the patterns for the heavy chain peptides of these subfragments are substantially similar but their light chain peptide patterns differ. The results suggest that the enzymatic and structural similarities that have been recognized between these isoenzymes using intact myosin hold true for the myosin subfragment-1. The differences between these subfragments are due to the differences in the light chains associated with them. PMID- 2975752 TI - Synthesis of functional bovine opsin in insect cells under control of the baculovirus polyhedrin promoter. AB - In vitro expression of cDNA encoding bovine opsin is accomplished using the baculovirus expression vector system. Full-length opsin was synthesized which was recognized by poly- and monoclonal antisera raised against bovine rhodopsin. Upon infection with a recombinant virus, 1 x 10(6) insect cells produced up to 3 micrograms opsin. Incubation of the in vitro synthesized opsin with 11-cis retinal produced a hydroxylamine-stable, photosensitive pigment. PMID- 2975753 TI - Adenovirus transcriptional regulatory regions are conserved in mammalian cells and Saccharomyces cerevisiae. AB - The adenovirus early region 3 (E3) promoter is an early viral promoter which is strongly induced by the adenovirus transactivator protein E1A. DNase I footprinting with HeLa cell extracts has identified four factor-binding domains which appear to be involved in basal and E1A-induced transcriptional regulation. These binding domains may bind TATA region-binding factors (site I), the CREB/ATF protein (site II), the AP-1 protein (site III), and nuclear factor I/CTF (site IV). Recently, it has been shown that the DNA-binding domain of transcription factor AP-1 has homology with the yeast transcription factor GCN4 and that the yeast transactivator protein GAL4 is able to stimulate transcription in HeLa cells from promoters containing GAL4-binding sites. These results suggest an evolutionary conservation of both transcription factors and the mechanisms responsible for transcriptional activation in Saccharomyces cerevisiae and higher eucaryotic organisms. To determine whether similar patterns of transcriptional regulation were seen with the E3 promoter in HeLa and yeast cells, the E3 promoter fused to the chloramphenicol acetyltransferase (cat) gene was cloned into a high-copy-number plasmid and stably introduced into yeast cells. S1 analysis revealed that similar E3 promoter mRNA start sites were found in yeast and HeLa cells. DNase I footprinting with partially purified yeast extracts revealed that four regions of the E3 promoter were protected. Several of these regions were similar to binding sites determined by using HeLa cell extracts. Oligonucleotide mutagenesis of these binding domains indicated their importance in the transcriptional regulation of the E3 promoter in yeast cells. These results suggest that similar cellular transcription factor-binding sites may be involved in the regulation of promoters in both yeast and mammalian cells. PMID- 2975754 TI - The amino-terminal region of the adenovirus serotype 5 E1a protein performs two separate functions when expressed in primary baby rat kidney cells. AB - Adenovirus serotype 5 E1a proteins immortalize primary cells and in cooperation with products of a second oncogene, such as adenovirus serotype 5 E1b or EJ ras, produce full transformation. E1a also activates transcription of specific viral and cellular promoters, represses enhancer-dependent genes, and induces cellular DNA synthesis in quiescent cells. Comparison of different adenovirus serotypes has identified three conserved regions in the E1a protein sequence. We have analyzed E1a mutants with deletions-linker insertions in or preceding the first conserved region, region 1 (amino acids 40 through 77 of adenovirus serotype 5 E1a). E1a mutants which have in-frame deletions-substitutions in region 1 or pre region 1 sequences were reconstructed into adenovirus to yield a total of 14 mutant viruses. All the mutant viruses showed wild-type growth in HeLa cells, confirming that region 1 is nonessential in these cells. However, we show that region 1 provides two distinct functions in infected primary rodent cells. One function is essential for induction of cell DNA synthesis, and the other is essential for focus formation. In addition, our results are consistent with a requirement for the DNA induction function in focus formation. PMID- 2975756 TI - Nutrition and somatomedin. XIX. Molecular regulation of insulin-like growth factor-1 in streptozotocin-diabetic rats. AB - Poor growth in diabetes involves low circulating levels of somatomedins/insulin like growth factors (IGFs), largely reflecting decreased growth factor release by the liver. To define regulatory mechanisms, circulating IGF-1 was compared with levels of a high mol wt putative hepatic IGF-1 precursor and hepatic IGF-1 mRNA in a model of progressive severity of diabetes in rats. Streptozotocin administered at 36, 72, 144, and 288 mg/kg produced graded metabolic decompensation 2 days later, from minimal hyperglycemia with continued weight gain at 36 mg/kg, to marked hyperglycemia, ketonemia, and weight loss at 288 mg/kg (all P less than 0.001). Total serum IGF-1 measured by RIA was unchanged with the 36 and 72 mg/kg doses of streptozotocin (471 +/- 19 and 439 +/- 27 ng/ml, respectively, vs. 517 +/- 27 ng/ml in controls) despite serum glucose greater than 400 mg/dl. With streptozotocin 144 and 288 mg/kg, serum IGF-1 fell to 131 +/- 27 and 142 +/- 10 ng/ml, respectively (both P less than 0.005 vs. controls). Serum IGF-1 was correlated strongly with serum beta-hydroxybutyrate and body weight (r = -0.88 and 0.91, respectively, P less than 0.0001), and less strongly with serum glucose (r = -0.59, P less than 0.0002). Extractable hepatic content of a high mol wt form of immunoreactive IGF-1 (a putative precursor) was unchanged at the two lowest doses of streptozotocin (68 +/- 4 and 83 +/- 9 ngeq/g vs. 67 +/- 4 in controls), but decreased to 16 +/- 3 and 29 +/- 4 ng/g at the two highest doses (both P less than 0.001 vs. controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975755 TI - Mapping of cellular protein-binding sites on the products of early-region 1A of human adenovirus type 5. AB - The binding sites for the 300-, 107-, and 105-kilodalton cellular proteins which associate with human adenovirus type 5 E1A products were studied with E1A deletion mutants. All appeared to bind to the amino-terminal half of E1A products in regions necessary for oncogenic transformation. These results suggest that these cellular species may be important for the biological activity of E1A products. PMID- 2975757 TI - Structural and functional relationships among receptors and regulators of the complement system. AB - The classical and alternative pathway of complement activation are regulated by a series of fluid phase and cell-bound factors, some of which at the same time serve as receptors for fragments of C3 and C4. These molecules are factor H, CR1 (C3b/C4b receptor), CR2 (C3d/EBV receptor), C4BP (C4b binding protein), DAF (decay accelerating factor), MCP (membrane cofactor protein; earlier designated p45/70), CR3 (iC3b receptor or Mac-1) and CR4 (protein 150/95). Due to structural, genetic and functional features these factors are members of one or several newly recognized large families of proteins: (1) molecules with 60 amino acids long repeats (H, CR1, CR2, C4BP, DAF); (2) proteins with 1,2-diacylglycerol membrane anchoring (DAF); (3) proteins with a heterodimer structure and preference for ligands containing the tripeptide arginine-glycine-asparagine (CR3, CR4). Recognizing the above mentioned regulators and receptors of the complement system as belonging to these protein families opens new perspectives for further genetic and functional research of mutual interest to complement and noncomplement scientists. PMID- 2975758 TI - Lysis of CR2-carrying cells by natural killer cells and by activated T lymphocytes is enhanced by C3 fragments. PMID- 2975759 TI - Regulatory effects of IgG-BF on hybridoma B cells. molecular characterization of variant cell lines. AB - Immunoglobulin G-binding factors (IgG-BF) produced by mouse T cells or hybridoma T cells (T2D4) were used to manipulate in vitro mouse hybridoma B cells. Both IgG production by, and proliferation of, these cells was inhibited by IgG-BF, or during co-cultures with IgG-BF-producing T2D4 cells. Thus, treatment of tumor B cells, besides its potential therapeutic use, represents an invaluable model for studying the regulation of Ig production by IgG-BF at a molecular level. To further analyze the molecular events induced by IgG-BF in B cell hybridomas, a set of variant clones of a hybridoma cell line (UN2) was isolated and variants were characterized for their Ig production and their Fc gamma R expression. PMID- 2975761 TI - Effects of various forms of monoclonal anti-Fc gamma R II (2.4G2) on B lymphocyte responses. AB - The effects of monoclonal anti-Fc gamma R II (2.4G2) in various forms on B lymphocyte proliferation and antibody (mu) secretion in vitro were evaluated. Soluble native 2.4G2 did not stimulate or inhibit the responses of B lymphocytes, either unstimulated or stimulated with rabbit F(ab')2 anti-mouse mu (anti-mu) plus lymphokines (the supernatant of Concanavalin A stimulated rat spleen cells). The failure of native 2.4G2 to affect responses was observed over a broad range of concentrations, and even when the B lymphocytes were incubated with the 2.4G2 for 24 hr prior to stimulation with anti-mu and lymphokines. Similarly, soluble chemically cross-linked 2.4G2 failed to affect B lymphocyte responses. Binding studies indicated that this failure was not due to a lack of binding and suggested that the polymerized 2.4G2 was cross-linking at least four Fc gamma R II. Larger multimers of 2.4G2 could not be evaluated due to a loss of binding activity. In contrast to the above results, 2.4G2 which was capable of extensively cross-linking Fc gamma R II (2.4G2 bound to Sepharose) or of cross linking Fc gamma R II to surface IgM (2.4G2 hetero-cross-linked with anti-mu) specifically inhibited B lymphocyte responses to anti-mu and lymphokines. Antibody secretion was affected more than proliferation. These results provide additional evidence that Fc gamma R II regulate the responses of B lymphocytes, and suggest that cross-linking of more than four Fc gamma R II is necessary to generate the inhibitory signal. Further, the results indicate that the ligand does not have to be internalized in order to generate the regulatory signal. Finally, the results with the heterodimer suggest that it may be possible to regulate a particular antibody response using anti-Fc gamma R II cross-linked to antigen or to anti-receptor antibody (e.g. an anti-idiotype). PMID- 2975760 TI - Molecular mechanisms regulating the expression of murine T-cell Fc gamma receptor II. PMID- 2975762 TI - Molecular recognition of antibody (IgG) by cellular Fc receptor (FcRI). AB - Earlier studies from this and other laboratories have provided indirect evidence for the involvement of the C gamma 2 domain of human IgG in the binding of IgG to the high affinity monocyte Fc receptor (FcRI). Two approaches have been used to extend these studies and to further localize the site of interaction on human IgG. Firstly, monoclonal antibodies (MAbs) directed against different epitopes on IgG were assayed for their capacity to inhibit the binding of radiolabelled IgG to human monocytes or U937 cells. The capacity of the MAbs to interact with their respective epitopes on FcR-bound IgG was also studied using indirect radiobinding and immunofluorescence assays. Secondly, a number of IgGs from several different species and fragments of human IgGs were assayed for their ability to inhibit the binding of radiolabelled IgG to human monocytes. The amino acid sequences of those IgGs exhibiting relatively tight, intermediate or weak binding to monocyte FcRs were compared. On the basis of these studies a possible monocyte FcR-binding site on human IgG is postulated, involving the lower hinge region of IgG (residues Leu 234-Ser 239) with possible involvement of the nearby N-proximal bend and two beta-strands (Gly 316-Lys 338). PMID- 2975763 TI - Inter-receptor relationships in effector cell triggering, with particular reference to the mast cell. AB - The role of IgE antibodies in the initiation of allergen induced release of mediators from sensitised mast cells is discussed; both from the point of view of their binding to the high affinity Fc(epsilon)RI receptor and in their provision of a triggering signal, resultant upon the cross-linking of cell bound IgE molecules by specific antigen (allergen). A possible inter-relationship between the Fc(epsilon)RI and a low affinity receptor for the IgG4 isotype on human mast cells is considered, in the light of evidence that the two types of receptor could be acting synergistically in certain clinical allergy situations (e.g. atopic eczema). It is suggested that a similar juxta-positioning of Fc(gamma 4)RII and Fc(epsilon)RI receptors on human B-lymphocytes might provide scope for regulation of IgE synthesis by IgG4 auto-antibodies directed against this immunoglobulin isotype. PMID- 2975764 TI - [Allergic contact eczema caused by Sargassum muticum in professional fishermen on Lake Grevelingen]. PMID- 2975766 TI - [Surface phenotypes of peripheral blood mononuclear cells in multiple sclerosis]. AB - Using monoclonal antibodies the subpopulations of mononuclear cells were studied in peripheral blood in patients with multiple sclerosis in active phase of the disease. The results suggested the following conclusions: 1. In the active phase of multiple sclerosis (during exacerbations and slowly progressive course) a rise occurs in the proportions of Ia-positive and M1-positive cells without changes in the proportions of T4 and T8 lymphocytes and in their mutual ratio. 2. No differences were observed in the percent of peripheral blood mononuclear cells between all patients in the active phase of the disease and those with slowly progressing disease. 3. During immunomodulatory treatment (alternating administration of prednisone and levamisole) a rise was observed in the percent of suppressor cells T8/Ia-positive, and a fall in the T4/T8 suppressor cell ratio. PMID- 2975765 TI - Diagnosis and analysis of the clinical course of T-phenotype acute lymphoblastic leukemia. AB - We refer to the diagnostic possibilities in T-phenotype acute lymphoblastic leukemia. Immunophenotyping based on the proof of T-differentiation antigens by means of specific monoclonal antibodies and assessment of the E receptor, as well as examination of enzymatic activities of adenosine deaminase and purinenucleoside phosphorylase, were performed. Thirty patients with acute lymphoblastic leukemia of T-phenotype were examined. The monoclonal antibodies used demonstrated a heterogeneity in this type of leukemia reflecting the stage of thymocyte differentiation. The cells of some patients with T-phenotype acute lymphoblastic leukemia expressed simultaneously the common acute lymphoblastic leukemia antigen and Ia molecules. Examination of enzyme activities demonstrated a characteristic pattern with a significantly increased adenosine deaminase activity and simultaneously decreased purinenucleoside phosphorylase activity in T-phenotype acute lymphoblastic leukemia. The unfavorable course of disease in patients with T-phenotype acute lymphoblastic leukemia was demonstrated analyzing a group of 50 children with acute lymphoblastic leukemia. By means of statistical methods the cumulative rate of children surviving with acute lymphoblastic leukemia of common and T-phenotypes have been expressed. The results showed the need for a more effective treatment aimed at the T-phenotype of acute lymphoblastic leukemia. PMID- 2975767 TI - [A case of mitochondrial enzymopathy]. AB - A case of mitochondrial enzymopathy, called also ophthalmoplegia plus, was observed in a 31-year-old man. Histoenzymatic investigations demonstrated in the myocytes decreased and irregularity of reactions for succinic dehydrogenase, tetrazole reductase and mitochondrial ATPase. In electron microscopy paracrystalline structures, lamellar bodies and concentrically condensed cristae were seen in the mitochondria, and increased glycogen stores outside the mitochondria. PMID- 2975768 TI - Histaminergic regulation of prolactin secretion: involvement of tuberoinfundibular dopaminergic neurons. AB - It has been shown that histamine (HA) stimulates prolactin (PRL) secretion via H2 receptors following intra-cerebroventricular infusion and via H1 receptors following systemic (intra-arterial) infusion. Since the effect of HA appears to be exerted at a suprapituitary level, we investigated the involvement of the tuberoinfundibular dopaminergic (TIDA) system in HA-induced PRL secretion in urethane-anesthetized male rats. HA infused intracerebroventricularly (30 micrograms) or intra-arterially (420 micrograms) decreased the dopamine (DA) concentration in pituitary portal blood by 30 and 23%, respectively. Blockade of DA receptors by pimozide did not prevent the stimulation of PRL secretion induced by intracerebroventricular infusion of HA or the H2 receptor agonist dimaprit. Furthermore, during DA receptor blockade intracerebroventricular infusion of the H1 receptor agonist 2-thiazolylethylamine inhibited PRL secretion. In contrast, pimozide prevented the stimulation of PRL secretion induced by intra-arterial infusion of HA and the H1 receptor agonist 2-thiazolylethylamine. In fact, under these conditions intra-arterial infusion of HA or the H2-receptor agonist dimaprit inhibited PRL secretion. During treatment with alpha-methyl-p-tyrosine, which reduced the hypothalamic DA content by 50%, HA infused intracerebroventricularly stimulated PRL secretion, while HA infused intra arterially inhibited the secretion, which is in accordance with the results obtained during pimozide treatment. Cholinergic blockade by atropine did not prevent the HA-induced PRL release, excluding the possibility that the observed effect of pimozide is due to its anticholinergic property. We suggest that intracerebroventricular infusion of HA by activation of H2 receptors may stimulate PRL secretion partly via inhibition of the TIDA system and partly via other mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975769 TI - Involvement of D1 dopamine receptors in the nicotine-induced neuro-endocrine effects and depletion of diencephalic catecholamine stores in the male rat. AB - Male rats were treated acutely with nicotine (4 x 2 mg/kg, 30-min time intervals, total treatment time 2 h) or exposed to cigarette smoke from 4 x 1 cigarette (30 min time intervals, total treatment time 2 h). Some rats were pretreated with the D1 dopamine (DA) receptor antagonist SCH 23390 (0.1-3.0 mg/kg, i.p.), or with the D2 DA receptor antagonists remoxipride and raclopride (1 mg/kg, i.p.), or with the 5-hydroxytryptamine 2 (5-HT2) receptor antagonist ketanserin (0.3 mg/kg, i.p.) 5 min before nicotine treatment or the acute intermittent exposure to cigarette smoke. Some rats were treated with the D1 DA receptor agonist SK&F 38393 (1-10 mg/kg, i.p.) 15 min, 30 min or 2 h before decapitation. Hypothalamic and pre-optic catecholamine (CA) levels were measured by quantitative histofluorimetry in discrete DA and noradrenaline (NA) nerve terminal systems. Serum thyroid-stimulating hormone (TSH), prolactin, luteinizing hormone (LH), follicle-stimulating hormone (FSH), vasopressin, corticosterone and testosterone levels were determined by radioimmunoassay procedures. Nicotine treatment and to a minor degree also acute intermittent exposure to cigarette smoke produced a reduction in serum prolactin, LH and TSH but not in serum FSH, vasopressin and testosterone levels. Nicotine treatment also increased serum corticosterone levels. Pretreatment with the D1 DA receptor antagonist SCH 23390 (1-3 mg/kg) counteracted the lowering of serum LH, but not of prolactin and TSH levels induced by nicotine or exposure to cigarette smoke. SCH 23390 alone (1-3 mg/kg) increased serum TSH levels. Remoxipride, raclopride or ketanserin did not counteract any of the neuro-endocrine actions induced by nicotine treatment. However, ketanserin alone lowered serum prolactin levels. SK&F 38393 increased serum TSH, prolactin and LH levels. It was found that nicotine treatment and exposure to cigarette smoke with few exceptions produced a depletion of CA stores in NA and DA nerve terminals of the hypothalamus, pre-optic area and median eminence which was counteracted by SCH 23390 (1 mg/kg) but not by remoxipride, raclopride (1 mg/kg) or ketanserin (0.3 mg/kg). The results indicate that D1 but not D2 DA or 5-HT2 receptors may modulate the NA and DA release in the median eminence, the hypothalamus and the pre-optic area induced by nicotinic cholinoceptor activation. Furthermore, D1 DA receptors in the median eminence may at least in part mediate the inhibitory effects of nicotine on LH but not on TSH and prolactin secretion, although there appears to exist a D1 DA receptor in the median eminence which inhibits TSH secretion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2975770 TI - Progressive myopathy in trisomy 21. AB - A 23-year-old female patient with clinical manifestations typical of Down's syndrome progressively developed intensifying weakness of the proximal muscles from the age of 16 years. CK levels were distinctly elevated. Electromyography showed myogenic lesions and muscle biopsy a myopathic image with extensive fibre hypertrophy. Progressive sporadic myopathy in association with Down's syndrome has not been reported previously. PMID- 2975771 TI - [Role of gynecologic laparoscopy in the pelvic pain syndrome]. PMID- 2975772 TI - [Effects of the oral administration of a contraceptive on lipid balance and on the concentration of serum tocopherol]. PMID- 2975773 TI - [Effects of oral estrogen-progestin contraceptives on blood coagulation and assessment of the risk of thromboembolism]. PMID- 2975774 TI - Effect of reduction of blood volume on plasma immunoreactive atrial natriuretic factor concentrations in normal man. AB - Plasma immunoreactive atrial natriuretic factor (i-ANF) concentrations were measured in ten voluntary blood donors before and after acute blood loss induced after they had been supine for 20 min. Lying down caused i-ANF to increase significantly in all subjects. Bleeding (400 ml in 5 min) caused i-ANF to decrease to values significantly lower than either those observed before haemorrhage or those recorded in the upright position. These results provide evidence that in normal man acute changes in the intravascular volume, although limited within physiological limits, are able to induce opposite changes in i-ANF release. PMID- 2975775 TI - [Effect of viral infections on the course of pregnancy and the health status of the infants]. PMID- 2975776 TI - [Experience with the mass use of an oral rehydration method in acute intestinal infections in children in a hot climate]. PMID- 2975777 TI - [Epidemiology of food intolerance in preschool children from urban and rural areas]. PMID- 2975778 TI - [10-year activities of the Leningrad inter-regional pediatric pulmonological center]. PMID- 2975779 TI - UVB reduces the cutaneous cellular infiltrate of atopic eczema: a preliminary study. PMID- 2975780 TI - Pseudoallergic reaction from Khellin in photochemotherapy of vitiligo: a case report. PMID- 2975781 TI - The evaluation of the gingival fluid flow meter. PMID- 2975782 TI - Transabdominal chorionic villus sampling: analysis of 350 consecutive cases. AB - We report a series of 350 patients submitted to transabdominal chorionic villus sampling (CVS). A technique using two ultrasound-guided needles and a suction pump was used. In most cases, the procedure was performed between 9 and 13 weeks. Twenty-one pregnancies were selectively terminated; nine spontaneous abortions followed the procedure and one fetal loss after 28 weeks was recorded; 153 pregnancies are in progress and 169 delivered fetuses are alive and well. Transabdominal biopsy is a feasible and effective technique for CVS. PMID- 2975783 TI - [Effect of preventive and social factors on the morbidity of children with local forms of tuberculosis]. PMID- 2975784 TI - Effects of gossypol on the activity of rabbit kidney Na,K-ATPase and the functions of human erythrocyte membrane. PMID- 2975785 TI - Modification of epsilon-amino group of lysines, cholesterol oxidation and oxidized lipid-apoprotein cross-link formation by porphyrin-photosensitized oxidation of human low density lipoproteins. PMID- 2975786 TI - Oxidative deamination of Se-(1-carboxyethyl)-,Se-(1-carboxypropyl)- and Se-(2 carboxyethyl)-selenocysteine by snake venom L-aminoacid oxidase. AB - Details are reported for the synthesis of Se-(1-carboxyethyl)-selenocysteine (1 CESeC), Se-(1-carboxypropyl)-selenocysteine (1-CPSeC) and Se-(2-carboxyethyl) selenocysteine (2-CESeC). They can be obtained in pure cristalline form with good yield. Some chromatographic properties, useful for their identification, are described. The three aminoacids are good substrates for snake venom L-aminoacid oxidase, giving the corresponding alpha-ketoacids as reaction products. PMID- 2975787 TI - Effects of phenytoin and flunarizine on the rise in extracellular potassium induced by repetitive stimulation of rat cerebral cortex. AB - At a critical intensity of repetitive stimulation, [K+]0 in cerebral cortex increases rapidly to a "ceiling" value of 10-12 mM. Phenomena related to this rapid regenerative increase in [K+]0 were investigated using K+-sensitive microelectrodes. The critical intensity required to induce this rise was found to be increased by phenytoin (90 mg/kg preload + 30 mg/kg presurgery) but not by flunarizine (30 mg/kg preload + 10 mg/kg presurgery), when the stimulus (20 Hz, 0.4 ms pulse duration, 5 or 40 s train duration) was applied to a 1 mm2 area of parietal neocortex in alfentanil-anesthesized rats. Flunarizine and phenytoin appear to differ in their mechanisms of anti-epileptic action. PMID- 2975789 TI - Brain interstitial fluid calcium concentration during development in the rat: control levels and changes in acute plasma hypercalcaemia. AB - Age-related changes in brain interstitial fluid (ISF) ionic calcium, in ionic and total calcium in plasma and the effect of plasma hypercalcaemia on ISF calcium have been studied in rats aged between late gestation and adult. ISF ionic [Ca2+] decreased significantly with development from 1.6 mM to 1.2 mM. Plasma ionic [Ca2+] was not significantly different from ISF [Ca2+] apart from a transient hypocalcaemia at birth which was not reflected in the ISF. Plasma total calcium was around 2X ionic [Ca2+] and showed the same age-related decrease. In acute plasma hypercalcaemia induced by calcium gluconate injections, there was only weak regulation of ISF Ca2+ at 21 days gestation but a rapid improvement after birth resulted in excellent control by 5 days. PMID- 2975788 TI - Stimulation-evoked changes in extracellular pH, calcium and potassium activity in the frog spinal cord. AB - Double-barrel ion-sensitive microelectrodes were used to measure activity-related changes in extracellular pH (pHe), potassium and calcium concentration ([K+]e and [Ca2+]e) in the spinal dorsal horns of frogs. Repetitive stimulation (30-100 Hz) of the dorsal root evoked transient acidification in the lower dorsal horn by 0.25 pH units, which was accompanied by an increase in [K+]e by 4-5 mmol/l and a decrease in [Ca2+]e by 0.5 mmol/l. The pHe changes were found to have a typical depth profile and increased with the stimulation frequency, intensity and duration. The maximum of pHe changes was reached in 25-30 s of stimulation, and when stimulation continued further no greater pHe changes were achieved. Similarly as the K+ and Ca2+ transients, the pHe reached a ceiling level, which was 0.2-0.25 pH units more acid than the pH of the Ringer solution. The poststimulation K+ undershoot below the resting K+ level (3 mmol/l) was accompanied by an alkaline shift before the original pH base line. The rise time of the pHe changes was slower than that of [K+]e and [Ca2+]e changes. However, the redistribution of all the ionic changes had a similar time course. The clearance of changes in [K+]e and pHe was slowed by ouabain. The depression of the acid shift required higher concentrations of ouabain than the depression of the alkaline shifts. Acetazolamide, a carbonic anhydrase inhibitor, depressed the acid and enhanced the alkaline shift. Superfusion of the cord with elevated [K+]e was accompanied by a prompt and progressive acid shift, the lowering of [K+]e by an alkaline shift. The stimulus-evoked K+ increase and acid shift were depressed during the elevated [K+]e, while the alkaline shift was enhanced. Spontaneous elevations of [K+]e were accompanied by acid shifts of a similar time course. The results are discussed in terms of stimulus-evoked changes in extracellular strong ion differences [SID]e, and of their possible physiological significance. PMID- 2975790 TI - Changes in [K+]0 induced by transretinal currents in frog retina. AB - Current-induced changes in extracellular potassium concentration (delta [K+]0) were measured with K+-selective microelectrodes in frog retina. A characteristic depth profile of delta[K+]0 was detected, which included significant changes in the superfusate just above the inner limiting membrane. This response was almost eliminated by Ba2+, a blocker of K+ channels. The delta[K+]0 in the superfusate is likely due to transcellular flux of K+, probably through Muller cell endfeet. PMID- 2975791 TI - The time course of the light-induced extracellular potassium change around receptors and at the vitreal surface compared with the time course of slow PIII wave in the isolated rabbit retina. AB - The experimental preparation was a piece of isolated rabbit retina, the lower side of which was superfused in a chamber with a plasma-saline mixture kept at 35 degrees C to which, in some experiments, either sodium aspartate and glutamate, or Mg2+, were added to suppress second order neuronal activity. The upper side was exposed to an atmosphere of humidified oxygen. With longlasting light stimuli (30-180 s) the [K+]0 around receptors first falls from its dark-adaptation value (4.6 +/- 0.4 mM) to 3.3 +/- 0.6 mM and then reaccumulates nearly to the pre stimulus value. The time course of reaccumulation varies greatly (30-180 s) between preparations. The peak time (90% value) of the [K+]0 decrease is several seconds. It seems that the more rapidly the retina is prepared the shorter the peak time of the [K+]0 reaccumulation and the more rapid the [K+]0 reaccumulation. This suggests that the [K+]0 reaccumulation is effected by active, oxygen-dependent mechanisms. PIII always precedes the [K+]0 change, its peak time is several 100 ms. During longlasting light stimuli (30-180 s) slow PIII reflects the difference in the [K+]0 changes around the photoreceptors and at the vitreal surface. The rapidly and carefully isolated preparations exhibit a slow cornea-positive potential similar to a c-wave, when second order neuronal activity is not suppressed. PMID- 2975792 TI - The effects of cetrimide and potassium bromate on the potassium ion concentration in the inner ear fluid of the guinea-pig. AB - The mammalian inner ear is located deep within the temporal bone. The organ of Corti, the delicate sensory system for sound, is surrounded by two fluid systems; the potassium-rich endolymph and the sodium-rich perilymph. The pathogenesis of inner ear deafness is thought to be largely due to an imbalance of potassium and sodium ions in the inner ear fluids. Dynamic changes in K+ in the endolymph and perilymph were studied in the guinea-pig following cetrimide (cetrimonium bromide, a powerful cationic detergent which shows ototoxicity) applications on the round window membrane, intramuscular injection of potassium bromate (bread whitener, known to cause renal damage and permanent deafness in animals and man). Maximum fall in K+ concentration in the endolymoh (mM/min) and maximum K+ conductance (mM/min/mV) were 3.54 +/- 1.65 and 0.036 +/- 0.02 in cetrimide, and 1.85 +/- 0.35 and 0.021 +/- 0.009 in potassium bromate, respectively. In view of these findings, the influence of the active transport mechanism to K+ concentrations are discussed in comparison with dynamic changes in endolymph K+ induced by asphyxia and ethacrynic acid. PMID- 2975793 TI - In vivo electrochemical studies of the effects of cocaine on dopamine nerve terminals in the rat neostriatum. AB - In vivo electrochemical measurements, involving chronoamperometric recordings using monoamine-selective Nafion-coated electrodes, were used to study the effects of locally applied cocaine (50-500 micromolar barrel concentrations) on dopamine (DA) nerve terminals in the neostriatum of the anaesthetized rat. Local application of cocaine did not elicit detectable increases in basal levels of extracellular DA. However, locally applied cocaine significantly augmented the concentration of DA detected following a potassium (K+)-evoked depolarization. Data obtained with a new high-speed chronoamperometric recording technique further support that DA is the predominant species detected electrochemically following potassium-evoked depolarizations both before and after local application of cocaine. Unlike other locally applied uptake inhibitors that we have studied, cocaine failed to augment the time dynamics of released DA. In addition, large doses of the highest concentration of cocaine caused an attenuation of K+-evoked DA release, presumably due to cocaine's local anaesthetic properties. These data suggest that cocaine elevates synaptic levels of DA, but in a manner that is not identical to other potent monoamine uptake inhibitors. PMID- 2975794 TI - Actions of calcitonin gene-related peptide on rat sensory ganglion neurones. AB - The membrane actions of calcitonin gene-related peptide (CGRP) and the effect of CGRP on the Ca-dependent action potential of rat dorsal root ganglion (DRG) neurons have been studied by means of an intracellular recording technique in isolated DRG of 2-3-week-old rats in vitro. Bath application of CGRP (10(-8)-10( 6) M for 1-5 min) elicited a slow reversible hyperpolarization and this hyperpolarizing effect was still observed in the medium containing TTX and TEA. However, about half of the large cells, classified by duration of action potential, were depolarized by CGRP. These membrane effects of CGRP were associated with an increase in membrane input resistance (about 20%). In addition, CGRP increased the duration of Ca-dependent action potentials. Our results are consistent with the role of CGRP as an excitatory neurotransmitter or neuromodulator in DRG-spinal cord. PMID- 2975795 TI - Scanning electron microscopic observations of the ependymal cell and the subependymal layer of the third brain ventricle in the rat. AB - This investigation was undertaken to clarify the three dimensional ultrastructure of the subependymal layer in relation with the ependymal cell layer in rat brain using the scanning electron microscope (SEM). The subependymal layer existing below the ependyma of the third ventricle in the brain of mature albino rats was examined with S E M. The hypothalamus freshly excised after median sagittal section was treated by collagenase with or without trypsin for a short while to remove the ependymal cells at the ventricular wall. After the enzymatic pretreatment of the specimen, many ependymal cells were removed and the subependymal layer was partially exposed. Most of the ciliated ependymal cells remaining at the ventricular wall extended long, single basal processes which then penetrated into the subependymal layer. The subependymal layer was composed of a delicate framework of thin processes of glial cells, ependymal cells and, in addition nerve cells. Scattered among the neuropil just beneath the ependymal cell layer, there were relatively small, globular subependymal cells. Occasionally, there were large bundles of unmyelinated nerve fibres in the subependymal layer. The individual nerve fibres distinctly showed many axonal varicosities within the fibres. Intermingled with the nerve fibres, glial processes of various forms were present. The structure of the ependymal cells and the subependymal layer was compared with the findings already reported in the studies using light and transmission electron microscope. PMID- 2975796 TI - Angiotensin II, bombesin and naloxone in tooth pulp and cutaneous nociceptive mechanisms in rabbits. AB - The effects of angiotensin II (A II), bombesin (B) and naloxone (N) on the amplitude of the late component of the evoked potentials of the cortex (EP) were studied by electrocutaneous (ECS) and tooth pulp (ETS) stimulation. An intraventricular (50 ng/kg) or intravenous (5 micrograms/kg) injections decreased the amplitude of the negative-positive component with a 20-40 ms latency (NP20 40) EP to ETS, but not to ECS. Saralasin (A II antagonist) injected intraventricularly (130 ng/kg) abolished this effect of A II in response to ETS. N intraventricular injections (30 micrograms/kg) increased the amplitude of NP20 40 EP to ETS, but not to ECS. B (intraventricular injection 20 ng/kg decreased the amplitude of NP20-40 EP to ECS, but not to ETS. This suggests there are the specific mechanisms in analgesia induced by A II and B in the pathways activated by ECS and ETS. PMID- 2975797 TI - Functional recovery after olfactory bulbectomy in rats: effect of embryonal brain grafts. AB - Regeneration of olfactory receptor neurones after bulbectomy can lead to formation of extrabulbar synapses, the functional significance of which remains controversial. Adult hooded rats (n = 27) were bilaterally bulbectomized under pentobarbital anaesthesia. Small pieces of brain tissues (1-2 mm3; OB: olfactory bulb; St: corpus striatum) were obtained from embryos of the same strain and placed into the bulbectomy-produced cavity in contact with the exposed brain surface. Smell was tested at 2- to 3-week intervals from 7 weeks to 7 months after the operation. The latency to find hidden food gradually improved and attained the intact control level in bulbectomized rats without grafts, but remained poor in the OB and St transplanted groups. Seven to ten months after transplantation, spontaneous unit activity and unit reactions to amyl acetate vapours were examined with a carbon fibre microelectrode. Unit responses in the transplants resembled those in the normal OB, but were less pronounced. Morphological examination of the transplant and of its connections with the olfactory receptor neurones and with the host brain suggested that functional recovery was mediated by the connections of the olfactory axons with the remnants of the OB, with the anterior olfactory nucleus and/or with the frontal cortex. The adverse effect of OB and St transplants was probably due to their poor connectivity with the host brain which prevented the regenerating olfactory axons from reaching higher olfactory centres. PMID- 2975798 TI - Epileptic phenomena produced by kainic acid in laboratory rats during ontogenesis. AB - Because of its preferential neuroexcitatory effects on the hippocampal neurones kainic acid (KA) is used for inducing partial seizures with a complex symptomatology. In this study the authors investigated the effect of intraperitoneal administration of KA, in doses of 2-16 mg/kg, on the laboratory rat during ontogenesis. The experimental animals were males aged 7, 12, 18, 25 and 90 days. The first signs of an effect in adult rats were automatisms; in young animals, jerks also appeared. The most important automatisms were wet dog shakes, which preponderated in 25-day-old and older animals, whereas in the young rats they consisted chiefly of intensive scratching. Minimal seizures with a motor pattern identical to minimal metrazol seizures were observed in all the age groups and so were generalized tonic-clonic convulsions, which appeared after large doses of KA. The systemic administration of KA is a convenient model of temporal seizures and their progressive generalization and could act as a model for testing broad spectrum antiepileptics. PMID- 2975799 TI - Electrophysiological properties of the isolated vestibulocerebellar complex of the frog in vitro. AB - Parameters of the electrical activity of the isolated vestibulocerebellar complex of the frog were studied under in vitro conditions. In the region of the vestibular nucleus (nc. VIII), in the presence of stimulation of the stato acoustic nerve (n. VIII), responses from efferent vestibular neurones and from unidentified (probably vestibulospinal) neurones were recorded. The latent periods of their excitatory postsynaptic potentials (EPSPs, 1.4-2.2 ms) were indicative of mono- and disynaptic connection. Inhibitory postsynaptic potentials (IPSPs) were also observed. Stimulation of the auricular lobe of the cerebellum evoked monosynaptic IPSPs, an EPSP-IPSP complex or pure EPSPs in nc. VIII, the latter probably by way of collaterals to the cerebellum. The inhibitory character of the effect of efferents from the cerebellum to the neurones of nc. VIII was demonstrated in the focal synaptic potential and in spontaneous and evoked unit activity. If n. VIII was stimulated, both focal and unit extra- and intracellular responses characteristic of activation of the Purkinje cells by mossy (MF) or climbing (CF) afferent fibres were recorded in the cerebellar cortex. The electrophysiological picture indicates that both synaptic transmission and the functional manifestations of the individual neurones are preserved in the tested preparation. PMID- 2975800 TI - Role of the medial preoptic area in sexual behaviour of the male rat: a study using repeated cycloheximide infusions. AB - The effect of cycloheximide (CHX), an inhibitor of proteosynthesis, on sexual behaviour was studied in adult male rats in which it was infused into the medial preoptic area of the hypothalamus (MPOA). Sexual interaction took place under control and modification of the precopulatory behaviour of the female. Among the various bilaterally infused amounts of CHX--20 micrograms in 1 microliter, 40 micrograms in 1 microliter, 80 micrograms in 2 microliters--the effect of the largest dose corresponded to a hypothetical state of copulatory readiness of the males; 2 h after administration the males were not capable of initiating copulatory behaviour with a passively receptive female, but did so successfully with a highly soliciting female. The dependence of copulatory readiness of the males on the precopulatory behaviour of the females was confirmed repeatedly after this dose of CHX. At the same time, the precopulatory activity of the males towards a passively receptive female was unimpaired. The effect of CHX was reversible; 48 h after infusion the males displayed high copulatory readiness. Only half the males (n = 7) given bilateral infusions of 80 micrograms CHX fulfilled criterion of copulatory performance. Histological control demonstrated that the MPOA was affected bilaterally by CHX infusion in 10 males out of 14. The results are discussed from the aspect of participation of the MPOA in the regulation of male sexual behaviour. The method allows changes in sexual behaviour to be studied in the same individual in a chronic experiment. PMID- 2975801 TI - Age-dependent influence of a moderate altitude (1,350 m) on the rat cardiopulmonary system. AB - The aim of the study was to determine whether a moderate altitude (1,350 m, Strbske Pleso, High Tatras) would act as a hypoxic stimulus on the cardiopulmonary system of young and adult rats. We used three experimental groups of animals differing in the duration of time for which they were kept at the given altitude (60 and 120 days) and the age at which they were acclimatized (from the 5th and the 60th day of life). The controls were kept at an altitude of 200 m (Prague). We found that an altitude of 1,350 m produced a significant increase in blood pressure in the lesser circulation; this response did not depend on the animals' age. Right ventricular enlargement occurred at the same time and was more pronounced in rats which had been acclimatized from infancy. The systemic blood pressure fell mildly, but significantly, only in animals exposed to altitude from adulthood; elevation of the haematocrit was likewise recorded only in this group. The results show that even a moderate altitude influences the cardiopulmonary system of the rat in a manner characteristic of the effect of chronic hypoxic hypoxia. PMID- 2975802 TI - A device for microcomputer controlled stimulation of cardiac muscle. AB - An interface is described which substitutes a stimulator for analysis of contractile properties of a muscle. A solution is presented for use of a computer both for analysis and stimulation control. A programme for such a purpose is described. PMID- 2975803 TI - Influence of age on the kinetics of vitamin C catabolism in guinea-pigs. AB - Pharmacokinetic analysis methods were used to evaluate the course of vitamin C catabolism in male guinea-pigs aged 3-5 weeks (weighting 255 g) and five months (weighing 570 g) given a diet containing no vitamin C. The vitamin C content of most of the organs and tissues of the younger experimental animals was depleted significantly more rapidly than in the older guinea pigs. At the same time, the half-time ascorbate elimination values were lower in the younger animals, in all the tissues analysed. As distinct from the older animals, vitamin C catabolism in the younger guinea-pigs had a biphasic character -- an extremely fast phase from the 1st to the 3rd day and a slower phase, comparable to the rate in older animals, from the 4th to the 17th day of administration of the scorbutogenic diet. The findings indicate that ascorbate metabolism in young organisms has specific characteristics, which ought to be taken into account when assessing and specifying the vitamin C requirements of the youngest section of the human population. PMID- 2975804 TI - Effect of the infusion of glucose, itralipid and nutramin on the initiation of rat liver regeneration after partial hepatectomy. AB - Rats were subjected to 67% hepatectomy and immediately after the operation were given a 4-hour infusion containing 5 ml saline solution, 28% glucose, 10% Intralipid, 8% glucose or 8% Nutramin. The rats were killed 18, 21, 24 and 30 h after partial hepatectomy. The effect of the tested solutions on the rate of liver regeneration was evaluated from changes in DNA specific activity and the mitotic activity of the hepatocytes. The infusion of 28% and 8% glucose markedly inhibited the onset of regeneration after partial hepatectomy. Nutramin likewise had an inhibitory effect, but not so pronounced as that of glucose. Conversely, the infusion of a lipid emulsion (Intralipid) and saline stimulated the initiation of liver regeneration compared with glucose and/or Nutramin. The possible mechanisms of the effect of infusion of the individual tested solutions on the onset of liver regeneration are discussed. PMID- 2975805 TI - Age-correlation of protein utilization to saccharide intake. AB - Male Wistar rats aged 30, 90, 150 and 360 days were fed ad libitum on diets with an optimum protein and fat content for their respective ages and an increasing saccharide content. Net protein utilization (NPU) was determined from the body nitrogen and protein intake values and the course of gluconeogenesis in the liver was measured by specific phosphoenolpyruvate carboxykinase (PEPCK) activity. According to the growth curve for the standard diet, animals aged 30 and 90 days have a high growth rate (3.245 g/day), 150-day-old rats grow more slowly (1.856 g/day) and 360-day-old animals put on scarcely any weight at all (277 mg/day). In 30-day-old rats, NPU attains maximum values in the presence of a 36% saccharide content in the diet, in 3- and 5-month-old animals in the presence of 51% saccharides and in one-year-old animals in the presence of 41% saccharides in their food. The course of gluconeogenesis also corresponds to these values. PEPCK activity in the youngest age group is greatest in the presence of 31% saccharides in the food, at 90 days it is stimulated in the presence of 31-46% saccharides, at 150 days the decisive concentration is 41 and 46% and at one year proteins are used for saccharide synthesis in diet with a 31 and 36% saccharide concentration. For optimum saccharide values, PEPCK activity is reduced in every age group; together with the maximum NPU values, this indicates that proteins are used for growth and building of the organism at an early age and for the renewal of tissues and organs and maintenance of the organism in adulthood. PMID- 2975806 TI - Antidepressant-induced weight gain: a comparison study of four medications. AB - Body weight change was monitored in 73 hospitalized depressed patients treated with one of four antidepressants for 1 month. After a 2-week medication-free period, patients were randomly assigned to treatment with amitriptyline, nortriptyline, desipramine, or zimelidine. By the end of 1 month, treatment with all three tricyclic compounds promoted weight gain, with the greatest increase observed during amitriptyline treatment; less weight was gained by patients treated with nortriptyline and desipramine. In contrast, most patients treated with zimelidine showed no weight gain and, in many cases, demonstrated weight loss. Weight change during treatment was not associated with age, sex, severity of depression, obesity, weight loss during depression, or clinical response. PMID- 2975808 TI - Modulation of cellular genes by oncogenes. PMID- 2975807 TI - Primary ciliary dyskinesia: cytological and clinical features. AB - Thirty patients with functional and/or morphological abnormalities of respiratory tract cilia were identified. The diagnosis of primary ciliary dyskinesia was based on observed abnormalities of ciliary ultrastructure or beating in vitro (beat pattern, beat frequency or percentage of motile cilia). Beat frequency and motility indices approached the normal range in some cases and suggests that the term 'immotile cilia syndrome' is not appropriate. Morphological abnormalities were most commonly due to deficiency of dynein arms, affecting the outer arms (n = 7), inner arms (n = 3) or both (n = 10). Examples of radial spoke and microtubular defects were also identified but in seven subjects ciliary ultrastructure was normal. In six patients paired samples of nasal and bronchial cilia were obtained and showed consistent abnormalities of motility and ultrastructure. Adenosine triphosphate and adenosine triphosphatase did not restore in vitro motility when added to dynein deficient cilia. The clinical picture was of life-long sinusitis and recurrent bronchial infection but the spectrum was broader than that encompassed by Kartagener's triad (dextrocardia, sinusitis and bronchiectasis). Fourteen patients had normal cardiac situs and definite or highly suggestive evidence of bronchiectasis was present in only 17 patients. Radiological evidence of sinusitis was common but absence of frontal sinuses was not universal. Chronic serous otitis media was a frequent finding but deafness was rarely profound. Fertility problems were common but were not universal in female subjects. Lung function testing revealed evidence of airflow obstruction but this was mild in most cases. PMID- 2975809 TI - Raclopride, a new selective ligand for the dopamine-D2 receptors. AB - 1. The use of raclopride, a new compound of the salicylamide series, as a ligand for the labelling of dopamine-D2 receptors in vitro and in vivo is described. 2. 3H-Raclopride has a high affinity for the dopamine-D2 receptors (Kd = 1 nM in rat striatum) with much less affinity for any other receptor. 3. 3H-Raclopride enters the brain easily and has therefore also been used in in vivo binding and autoradiography. The nonspecific binding is very low both in vitro and in vivo. 4. Raclopride has been labelled with 11C, and is used as a marker for dopamine-D2 receptors in the living human brain using positron emission tomography. PMID- 2975810 TI - Competition for sucrose-pellets in triads of male Wistar rats: effects of three serotonergic drugs. AB - 1. Within triads of rats, individuals can be discerned according to their competition for sucrose-pellets as a high-, a medium- and a poor-performing animal. 2. The competition-rates, expressed as scores, are affected by serotonergic drugs: upon inhibiting tryptophan-hydroxylase, the characteristic abstention of the poor-performing rats can temporarily be overcome; quipazine, on the other hand, leads to a dose-dependent decrease in the competition-rate of high-performing rats. 3. Such findings are indicative for a regulatory effect of some serotonergic mechanisms on a competition-behavior evoked within the familiar social context. PMID- 2975811 TI - Cingulotomy in the rat fails to block opiate withdrawal effects but elevates stress-induced plasma beta-endorphin. AB - 1. Male, albino, Sprague Dawley rats underwent surgical cingulotomy or sham operation and were then implanted with subcutaneous morphine pellets or exposed to forced running. 2. The conditions of cingulotomy or forced running alone did not cause a significant increase in plasma i.r. Beta endorphin concentrations. 3. The combined treatment of cingulotomy plus forced running caused a significant elevation of plasma beta-endorphin concentrations. 4. The cingulum is involved in the elicitation of stress hormone responses in the rat. PMID- 2975813 TI - [A word processor system for the visually handicapped]. PMID- 2975812 TI - [Chronic medullary neuro-stimulation in lumbosacral spinal arachnoiditis]. AB - Between 1983 and 1986, 20 patients were treated by chronic spinal cord stimulation for the relief of pain. These patients suffered from lumbar arachnoiditis or root fibrosis causing lumbar pain and sciatica following operations for disc herniation or repeated radiculography. In this short series, the results obtained were satisfactory, with 12 good results which were stable over a 2-year period, thanks to careful selection of the subjects within the framework of a multidisciplinary pain clinic. The treatment of the fibrosis and the mechanism of neurostimulation are reviewed. PMID- 2975814 TI - [Auricular natriuretic factor]. PMID- 2975815 TI - [Beta-lactamase-producing Neisseria gonorrhoeae and its antimicrobial susceptibility in Valdivia]. PMID- 2975816 TI - [An odontological study on Down's syndrome. Part 2: Crowding and gingivitis]. PMID- 2975817 TI - [Use of the computer in pedodontics. An application of the data base system as an aide in diagnosis]. PMID- 2975818 TI - [A case report of constitutional erythroid hypoplasia (Josephs-Blackfan-Diamond Type) with immunodeficiency]. PMID- 2975819 TI - [The dental research in institutions and schools for the handicapped in Osaka prefecture]. PMID- 2975820 TI - [Oral functional, developmental and hygienic problems, and their association in handicapped children. Part II. Oral findings in cerebral palsied children--the eruption of teeth, dental arches and occlusion]. PMID- 2975821 TI - [A survey on the contraction of dental diseases in handicapped children in our pedodontic outpatient clinic and our special clinic for handicapped children. 1st Report: 1st, 2nd and 3rd group according to Uehara's classification]. PMID- 2975822 TI - [Induction of HBs antibodies using a new genetically engineered vaccine]. PMID- 2975823 TI - [Structure of the basic rheumatic diseases of the adult population of Tbilisi]. PMID- 2975824 TI - Antibody response to heat-inactivated hepatitis B vaccine (CLB-3 micrograms) in hemodialysis patients and occupational risk personnel: a one year follow-up. PMID- 2975825 TI - [Research for the assessment of health needs of the pediatric population of a small local health district in Calabria]. PMID- 2975826 TI - [Venous thrombolysis and venous angioplasty]. AB - Venous thrombosis of the upper extremities was successfully lysed in 7 patients using selective infusion of urokinase. Selective lysis requires a lower dosage of urokinase and has a lower rate of side effects. Venous angioplasty was applied in 2 patients with stenoses of the subclavian vein, 3 with stenosis in dialysis shunts, and 1 patient with a long stenosis of the iliac vein. Venous angioplasty was successful and particularly useful in dialysis shunt stenoses. PMID- 2975827 TI - The efficacy of rifampicin against Staphylococcus aureus in vitro and in an experimental infection in normal and granulocytopenic mice. AB - The effect of rifampicin on Staphylococcus aureus in vitro was assessed as the difference between the logarithms of the numbers of colony forming units (CFU) with and without 3 h of exposure to the drug. The efficacy was expressed as the EC50, i.e. the concentration at which 50% of the maximal effect was obtained, calculated according to the Hill equation. The value found for the EC50 was 3.8 micrograms/l and the mean maximal effect was a log ratio of 5.03 (SEM 0.33). In vivo experiments were performed in normal mice and in mice made granulocytopenic by irradiation. The effect of rifampicin was assessed as the CFU count 5 h after the injection of a suspension of bacteria into the thigh muscle and 4 h after the administration of rifampicin. The efficacy was expressed as the ED50, i.e. the dose at which 50% of the maximal effect is obtained. This value was 0.18 mg/kg for the normal mice and 0.15 mg/kg for the granulocytopenic mice. The corresponding mean plasma concentrations of non-protein-bound drug were 28 and 24 mg/l, respectively. Thus, the EC50 was found to be much higher in vivo than that in vitro. This difference should be taken into account when parameters of in vitro efficacy are applied to establish dosage schedules. PMID- 2975828 TI - Tumor markers in bladder cancer. PMID- 2975829 TI - [Kinetic, cytogenetic, morphometric characteristics and DNA content of transitional cell bladder carcinomas]. PMID- 2975830 TI - Ureteral stone management: our experience. PMID- 2975831 TI - Ureteral "steinstrasse" after ESWL. PMID- 2975832 TI - Radical T.U.R. as a cure for prostatic carcinoma (stage A-B1)? An early evaluation of a temptative proposal. PMID- 2975833 TI - The use of transurethral and transrectal ultra-sounds in T.U.R. of prostatic tumours. PMID- 2975834 TI - The masked organic factors in impotence. PMID- 2975835 TI - Technique of dorsal penile artery bypass graft with saphenous vein for arteriogenic impotence. PMID- 2975836 TI - Physiology, hemodynamics and pharmacology of penile erection. PMID- 2975837 TI - [Therapeutic considerations in induratio penis plastica]. PMID- 2975838 TI - Iatrogenic lesions following ureteric dilatation. An experimental study. PMID- 2975839 TI - "Short-term" prophylaxis by aztreonam in prevention of post-operative infections in patients undergoing transurethral surgery. PMID- 2975841 TI - [Rhabdomyolysis following ligation of the internal spermatic vein in varicocele]. PMID- 2975840 TI - [Percutaneous endoureteral manipulation in the resolution of ureteral fistula complications following Bricker's urinary diversion: clinical case]. PMID- 2975842 TI - [Bladder neoplasm secondary to distant tumors: a case of bladder metastasis from gastric adenocarcinoma]. PMID- 2975843 TI - [Giant hydroureteronephrosis. Description of a case and review of the literature]. PMID- 2975844 TI - [Primary mixed cell renal adenocarcinoma following spinocellular carcinoma of the lung. Report of a rare case]. PMID- 2975845 TI - [The role of prevention in urology]. PMID- 2975846 TI - [Prevention and early diagnosis of prostatic carcinoma: results of mass screening]. PMID- 2975847 TI - [Significance of microhematuria in over 50 years of mass urologic screening]. PMID- 2975848 TI - [Prevention of nephrolithiasis]. PMID- 2975849 TI - [Prognostic factors in the tertiary prevention of superficial transitional carcinoma of the bladder]. PMID- 2975850 TI - [Role of prevention in pediatric urology]. PMID- 2975851 TI - [Prevention in andrology]. PMID- 2975852 TI - [Prevention and the neurogenic bladder]. PMID- 2975853 TI - [Urolithiasis; methods and results of endoscopic lithotripsy]. PMID- 2975854 TI - Second generation lithotripters: designs and clinical results reviewed. PMID- 2975856 TI - [Development and progress in the therapy of penile induration: 15 years' experience]. PMID- 2975855 TI - Second generation lithotripsy: an experience of 200 patients. PMID- 2975857 TI - Urethral calculi. PMID- 2975858 TI - Rigid operative cystoscopy without general anaesthesia. PMID- 2975859 TI - [Urinary sediment revisited: new aspects of an old test]. PMID- 2975860 TI - [Surgical therapy of stress urinary incontinence and detrusor instability]. PMID- 2975861 TI - [Extracorporeal lithotripsy (ESWL): 2 years' experience]. PMID- 2975862 TI - Diagnosis and treatment of early carcinoma of the prostate. PMID- 2975863 TI - Association of ABO (H) antigen negative transitional cell tumours with abnormal mucosal biopsies in superficial bladder cancer. PMID- 2975864 TI - [Ureteral invagination secondary to papilloma: an unusual cause of urinary tract obstruction]. PMID- 2975865 TI - [A variant of continent cystostomy according to Mitrofanoff of the neurogenic bladder]. PMID- 2975866 TI - [New perspectives on the use of Pygeum Africanum in prostato-bladder pathology]. PMID- 2975867 TI - Workers' compensation: 1980-84 benchmark revisions. AB - The first workers' compensation program was introduced 80 years ago. Its purpose was to compensate occupationally injured workers and their families for lost wages and medical expenses from job-related injury, regardless of fault. Today, each of the State and Federal programs that provides coverage to more than 86 percent of the work force uses a combination of private insurance, State or Federal funds, and self-insurance to meet its benefit obligations. The workers' compensation program is of continuing interest to the Social Security Administration (SSA) for several reasons. Since 1965, Social Security Disability Insurance benefits have been subject to reduction if such benefits, when combined with those provided under workers' compensation laws, exceed 80 percent of the worker's earnings. Because the two programs have gaps in protection as well as duplication in coverage, a periodic review of the workers' compensation program is necessary. In addition, SSA administers Part B of the Black Lung program- established to provide income-maintenance protection to coal miners disabled by pneumoconiosis--to about 1 million beneficiaries whose claims were filed before July 1973. This article provides revised benchmark data on the workers' compensation programs and presents a review of program operations during the early 1980's. PMID- 2975868 TI - [The role of hereditary factors in the development of psoriatic arthritis and the nature of its interrelations with psoriasis]. AB - Altogether 83 families of probands with psoriatic arthritis and the same number of families of probands with common skin psoriasis were investigated in order to study the role of hereditary factors in the development of psoriatic arthritis and the type of its interrelationship with psoriasis. Obvious accumulation of repeated cases of this disease was observed among the probands' relatives. Genetic analysis made it possible to reject the monogenic type of inheritance of psoriatic arthritis and to prove the multifactorial nature of its inheritance confirmed in 68%. Analysis of the distribution of repeated cases of psoriatic arthritis and uncomplicated psoriasis in both types of families helped to determine a genetic correlation coefficient (0.93) indicating great similarity of the genetic systems of both types of diseases therefore they could be regarded as the clinical variants of one disease--psoriasis. PMID- 2975869 TI - [Atrial natriuretic factor--a new hormone system]. PMID- 2975870 TI - Unusual activation mechanism of plasminogen. PMID- 2975871 TI - The levels of protein C and protein S in plasma in patients with type II diabetes mellitus. PMID- 2975873 TI - [Compensation practice of trade associations in relation to radiation-induced diseases: probability of causation]. PMID- 2975872 TI - Acetylsalicylic acid, BM 13.177 and picotamide improve the survival of endotoxin infused rabbits. PMID- 2975874 TI - [Changes in oral cavity in retarded immobilized children]. PMID- 2975875 TI - [Main currents of medical scientific theory in Germany after 1945]. PMID- 2975876 TI - [Organic life as circle movement: on the significance of the circle metaphor in F. W. J. Schelling's natural philosophy]. PMID- 2975877 TI - [Cocaine euphoria and natural science]. PMID- 2975878 TI - [The life and work of Martin Luther from the medical viewpoint--an attempt at interpretation]. PMID- 2975879 TI - [Urine analysis in the diagnosis of Isaak Judaeus, Gilles de Corbeil and Ortolf von Baierland. Observations on the processing technic]. PMID- 2975881 TI - Paravicius immortal? PMID- 2975880 TI - [Hematoscopy in German diagnosics of the late middle ages. Supplements to Friedrich Lenhardt, from the manuscript of Ortlof von Baierland's "Arzneibuch"]. PMID- 2975882 TI - National policies for hepatitis B vaccinations. PMID- 2975884 TI - [Reconstructive surgical treatment of invalids with defects of part of the tibial diaphysis]. AB - The authors have analyzed their observations of 113 patients and concluded that it is necessary to differentially use various kinds of osteosynthesis and bone autoplasty. Good and satisfactory long-term results were obtained in 94% of the patients operated upon. PMID- 2975883 TI - [The role of T-lymphocyte subpopulations in keratoplasty of burn leukoma]. PMID- 2975885 TI - [Locally destructive tumors]. PMID- 2975886 TI - [Significance of the initial immunologic status of patients in restorative operations on the large intestine]. PMID- 2975887 TI - [Prevention of postoperative eventration]. PMID- 2975889 TI - [Relaparotomy (problems of diagnosis and surgical tactics)]. PMID- 2975888 TI - [A method of suturing the laparotomy wound in postoperative eventration]. PMID- 2975890 TI - [Relaparotomy in the planned surgical treatment of peptic ulcer]. PMID- 2975891 TI - Intestinal permeability to macromolecules in piglets infected with transmissible gastroenteritis virus. AB - The permeability of the intestine of specific pathogen free piglets was investigated by measuring the concentration of 125-I in the blood after oral administration of 125-I polyvinylpyrrolidone (125-I PVP, MW = 40,000 Da) and the concentration of 131-I in the faeces after intravenous administration of 131-I porcine albumin (131-I PA, MW = 68,000 Da). The tests were performed one day before and up to two days after the piglets were infected with the Miller strain of transmissible gastroenteritis (TGE) virus. Biopsies of the jejunum were taken at the end of the experiment and blood samples were taken six-hourly. The piglets became anorexic and had diarrhoea 12 hours after infection; the packed cell volume decreased and the concentrations of urea and total serum proteins increased slightly after infection. However, the marked villous atrophy was not accompanied by an increased permeability of the intestine to PVP or PA. PMID- 2975892 TI - [Creative contribution of Poles to the surgical treatment of uterine prolapse]. PMID- 2975893 TI - [Surgical aspects of the treatment and significance of angina pectoris]. AB - Due to recent developments in modern interventional cardiology for the majority of patients with multi-vessel disease an indicatory and therapeutical grey-zone between bypass surgery and coronary angioplasty has arisen. Only patients with left main stenosis or uncomplicated one/two-vessel disease are generally accepted to be operated upon or treated by angioplasty, respectively. While pharmacological therapy is somewhat controlled by governments - surgical and cardiological interventional approaches are not. Even if it is justified to work out new interventional therapeutic approaches in the interest of its progress all of our current patients should in any case be informed concerning the risks of their treatment, prior to treatment. PMID- 2975894 TI - Compensatory changes of sympathetic tone, the renin-angiotensin-aldosterone system, vasopressin, and ANF as potential therapeutic targets in congestive heart failure. AB - Congestive heart failure (CHF) is not only reflected by such mechanical problems as forward- and backward failure, but it is also associated with a complex pattern of compensatory neuro-endocrine mechanisms, e.g., enhanced activity of both the sympathetic nervous system (SNS) and the renin-angiotensin-aldosterone system (RAAS); enhanced release of vasopressin from the pituitary gland; enhanced release of the atrial natriuretic factor (ANF) from the cardiac atria. These neuro-endocrine mechanisms not only operate as such but also display a complex pattern of mutual interactions. These mechanisms, though potentially beneficial short-term, may also be harmful when persisting during progression of the disease. For this reason they offer potential targets in the treatment of CHF besides the classical measures aimed directly at improving cardiac contractility. The following groups of drugs are discussed as therapeutic measures that suppress the aforementioned detrimental compensatory mechanisms: various types of vasodilator drugs; diuretics; beta 1-adrenoceptor blocking agents in low dosage; saralasin; ACE-inhibitors. So far, the enhanced release of vasopressin and ANF have not offered realistic new therapeutic targets in CHF, although their pathophysiologic issue is highly relevant. PMID- 2975895 TI - [Conditions of secretion and peripheral effects of atrial natriuretic peptide in heart failure]. AB - In heart failure, ventricular function is influenced by modulation of pre- and afterload. It has been shown that the secretion of atrial natriuretic peptide (ANP) is increased in experimental preparations of heart failure and in patients with cardiac failure. Distension of myoendocrine cells of the atria is the major stimulus for the release of ANP. The possible beneficial effect of ANP in heart failure consists of unloading the heart by its vasodilatory, diuretic, and natriuretic properties. Intravenous application of high, pharmacological doses of ANP improves ventricular performance by a reduction of pre- and afterload, the renal effects however, are markedly attenuated. From animal experiments one may conclude that ANP may play an important role in early and moderate heart failure as a counterregulating mechanism of the renin-angiotensin-aldosterone system. In chronic severe heart failure vasoconstrictory, volume, and sodium-retaining mechanisms override the effects of ANP, deteriorating ventricular function by an inadequate increase of pre- and afterload. The attenuation of the effects of ANP in cardiac failure may be due to altered hemodynamics especially in the kidney, to a counterregulation of vasoconstrictory and volume-retaining neurohumoral factors, to a reduction of specific binding sites for ANP (receptor "down regulation"), and to a possible intracellular defect beyond the formation of cGMP, the intracellular second messenger of ANP. PMID- 2975896 TI - [Morphologic characteristics of human skin in contact with organophosphate pesticides]. PMID- 2975897 TI - [The polymorphism of psoriasis in children and its relation to the HLA histocompatibility system]. PMID- 2975898 TI - [Acne fulminans]. PMID- 2975899 TI - [Petr Vasil'evich Kozhevnikov (on the 90th anniversary of his birth)]. PMID- 2975900 TI - [Bernardino Ramazzini--the founder of occupational dermatology]. PMID- 2975901 TI - Is the phosphofructokinase-reaction obligatory for glucose fermentation by Saccharomyces cerevisiae? PMID- 2975902 TI - Involvement of lipids in solute transport in yeasts. PMID- 2975903 TI - [The behavior of central venous pressure and acid-base balance in gynecologic laparoscopy]. AB - The anaesthetic procedure in patients affected with cardiac or other diseases during gynaecological laparoscopy has been reported. We tried with measurements of central venous pressure and control of acid-base-status to enable tese patients having laparoscopy, too. PMID- 2975904 TI - Granulocyte activating factor released from Propionibacterium acnes. A possible mediator of inflammation in acne vulgaris. AB - Incubation of Propionibacterium acnes but not of Propionibacterium granulosum or Propionibacterium avidum (for 30 min at 37 degrees C in physiological saline) released a soluble factor that produced enhanced chemiluminescence response of human granulocytes as well as increased chemotactic motility of these cells. Sephadex G-25 filtration of the granulocyte activating factor (GAF) revealed its low molecular weight and apparent peptide character. Thus, GAF may be a stimulus for inflammation in acne vulgaris since low molecular weight chemotactic factors can be expected to penetrate follicular walls. PMID- 2975905 TI - [History of the development of psychiatric care in Kursk Province]. PMID- 2975906 TI - [Periodic paramyotonic paralysis]. AB - A rare case of Eulenburg's paramyotonia and periodic paralysis combination is described in one family. The question of whether the disease presents a nosological entity is discussed with due consideration of its clinical polymorphism and the data of literature. Mapping the genes determining the disease would be decisive in this respect. PMID- 2975907 TI - Autologous immune response of tonsillar lymphocytes. AB - Tonsillar (TL) and peripheral blood lymphocytes (PBL) from patients with pustulosis palmaris et plantaris (PPP) or recurrent tonsillitis (RT) were analysed for their proliferative response of T-cells upon stimulation with non-T cells in the autologous mixed lymphocyte reaction (AMLR). TL and PBL from the same donor were cultured in the AMLR for up to 7 days. Maximum proliferation was observed after a 6-day culture in PBL, whereas that of TL was observed on the 3rd day of the culture. It was also observed that the AMLR of TL in patients with PPP was significantly (p less than 0.01) lower than in patients with RT. On the other hand, it has previously been proved by an immunofluorescence study that there is an identical antigenicity between tonsillar epithelium and skin. From this result, a blastoid transformation study of TL and PBL from patients with PPP was performed by using homologous or autologous skin extracts. Both homologous and autologous skin extract induced blastoid transformation of TL but PBL scarcely responded. These results may support the opinion that an auto-immunological mechanism may take part in the onset of PPP. PMID- 2975908 TI - Subsets of tonsillar lymphocytes and activated cells in each subset analysed by three-color flow cytometry. AB - Among the four subsets of tonsillar T cells, active cells were most abundant in the helper T subset, followed by the cytotoxic T, inducer T, and suppressor T subsets in order of decreasing cell count. The number of active cells was very small in the suppressor T subset. Plasma cells (PCA-1-positive cells) accounted for only about 3% of tonsillar cells. However, nearly half of the tonsillar B cells were activated (positive for Ba, being either Ba+/IgD+ or Ba+/IgD-). Ba positive cells were hardly found in the population of peripheral-blood B cells. Ba+/IgD+ cells, which are ready to be transformed to cyclic cells or dormant cells depending upon the degree of stimulation, outnumbered cyclic cells. Many Ba+/IgD+ cells were present even in the small tonsils of adults, although these cells were very scarce in the peripheral blood. These findings show that the tonsil is prepared to perform immunologic functions in prompt response to external stimuli. PMID- 2975909 TI - Effects of antidiuretic hormone (AVP) and human atrial natriuretic peptide (hANP) on water and electrolyte balance in neonates. PMID- 2975910 TI - Renal responses to intracerebroventricular infusions of atrial natriuretic peptide in the conscious goat. PMID- 2975911 TI - Plasma levels of beta-endorphin, cortisol, prolactin and growth hormone in depressed patients. AB - Basal serum cortisol, growth hormone, prolactin and immunoreactive (IR) plasma beta-endorphin levels were measured in 31 depressed patients (14 endogenous, 17 nonendogenous) undergoing the dexamethasone suppression test. The endogenously depressed patients had significantly higher (22.55 +/- 1.34 micrograms/dl) predexamethasone cortisol levels than the nonendogenous patients (16.34 +/- 1.93 micrograms/dl). The mean serum prolactin and growth hormone values of these two groups were not significantly different, while plasma IR-beta-endorphin levels of the endogenous group (40.11 +/- 3.57 pg/ml) were significantly lower than those of the nonendogenous group (120.33 +/- 27.98 pg/ml). Neither group showed a significant correlation between plasma IR-beta-endorphin and serum cortisol values. These results indicate that measurement of predexamethasone serum cortisol values and plasma IR-beta-endorphin could be valuable laboratory tests in the diagnosis of depression. PMID- 2975912 TI - Improvement in female patients with severe obsessions and/or compulsions treated with cyproterone acetate. PMID- 2975913 TI - Dehydroepiandrosterone and structural analogs: a new class of cancer chemopreventive agents. PMID- 2975914 TI - Changes in metastasis formation and distinct cell surface molecules in adhesion variants of a metastatic murine tumor. PMID- 2975915 TI - Regulation of the metastatic phenotype by the E1A gene of adenovirus-2. AB - We have previously demonstrated that rat embryo cells transformed by the ras oncogene alone are both tumorigenic and highly metastatic when injected into nude mice. In contrast, rat embryo cells transformed with the ras oncogene and the adenovirus 2 (Ad2) Ela gene are tumorigenic but either fail to metastasize, or exhibit a very low metastatic potential. Here we demonstrate that transfection of the Ad2 Ela gene into several of the ras transformed rat embryo cell lines results in a dramatic reduction in metastatic potential relative to the parental cell line. Transfection of cDNAs for the 12S and 13S Ela transcripts showed that both gene products are capable of reducing the metastatic potential of the ras transformed cell lines, however the 12S cDNA was more effective. This effect is specific to the Ad2 Ela gene as ras transformed cell lines expressing the Ad12 Ela gene or the human N-myc gene maintained their high metastatic potential. We hypothesize that the Ad2 Ela gene may regulate the expression of one or more cellular genes that contribute to the metastatic phenotype. PMID- 2975916 TI - Drug-induced tumor progression in a rat rhabdomyosarcoma. PMID- 2975917 TI - The role of pharmacokinetics in the clinical treatment of metastases. PMID- 2975918 TI - Measuring the serotonin uptake site using [3H]paroxetine--a new serotonin uptake inhibitor. AB - Serotonin is an important neurotransmitter that may be involved in ethanol preference and dependence. It is possible to label the serotonin uptake site in brain using the tricyclic antidepressant imipramine, but this also binds to other sites. We have used the new high-affinity uptake blocker paroxetine to define binding to this site and report it to have advantages over imipramine as a ligand. PMID- 2975919 TI - Discovery of novel brain lipoxygenase products formed from docosahexaenoic acid (22:6w3). AB - It is known that alcohol exposure leads to a decline in brain docosahexaenoate (22:6w3). We hypothesized that alcohol could stimulate the metabolism of this polyunsaturated fatty acid to bioactive products. Several oxidized products of 22:6w3 were indeed observed when rat brain homogenate was incubated with 14C22:6w3 in vitro. A similar group of metabolites was formed in vivo from 14C 22:6w3 injected into the lateral ventricle. These metabolites were characterized by thermospray- and GC/MS as well as by the synthesis of standards using purified enzymes. Platelet lipoxygenase also proved useful in identifying one of the brain metabolites and served as a source of enzyme for preparative studies. Their physiological effects on smooth muscle tone and platelet aggregation will be presented. PMID- 2975920 TI - Milk plasmin, antitrypsin, N-acetyl-beta-D-glucosaminidase and bacterial growth in lactoserum during the early post partum period. PMID- 2975921 TI - Ochratoxin A as a suppressor of mitogen-induced blastogenesis of porcine blood lymphocytes. PMID- 2975922 TI - Assessing support networks: stability and evidence for convergent and divergent validity. AB - Data on important methodological problems in assessing support networks is presented. Stability (over 1 year) and construct validity were examined in two groups of mothers of preschool children. One group of mothers had children with a physical handicap (n = 25); the other group had normally developing children (n = 44). When their children were 4 years of age and again at 5 years of age, mothers' support networks were assessed with the Pattison Psychosocial Kinship Inventory (a semistructured interview) and a log diary. Results indicated considerable stability in mothers' support networks over the 1-year period. In addition, agreement across methods was fairly high despite marked differences in format, providing evidence for the construct of support networks. Further evidence for construct validity was provided by generally low correlations between support network variables and other measures of social ecology. Findings were roughly similar for both groups of mothers indicating general replicability of findings. PMID- 2975923 TI - Parental coital rates and Down syndrome. PMID- 2975924 TI - Nondisjunction in Down syndrome. PMID- 2975925 TI - [Hemodynamic effects of fentanyl and sufentanil combined with flunitrazepam in coronary artery surgery]. AB - Because sufentanil has been reported as being able to prevent or treat peroperative hypertensive crises during aorto-coronary artery graft surgery, a study was carried out to compare the haemodynamic effects of sufentanil with those of fentanyl. 20 patients who were to undergo aortocoronary bypass grafting (CABG) were randomly allocated to two equal groups, sufentanil (Sf) and fentanyl (F) groups. A 1 to 5 dose ratio was used so as to have equipotent doses of sufentanil and fentanyl. Induction doses were 10 micrograms.kg-1 sufentanil and 50 micrograms.kg-1 fentanyl. Up to 20 micrograms.kg-1 sufentanil and 100 micrograms.kg-1 fentanyl were then used between intubation and the setting-up of cardiopulmonary bypass (CPB). A bolus of 10 micrograms.kg-1 flunitrazepam was given if necessary, so as to lower the mean arterial pressure (Pa) to below 100 mmHg after intubation, and under 80 mmHg during CPB. Heart rate, Pa, mean pulmonary arterial pressure, pulmonary wedge pressure (Ppw), central venous pressure and cardiac output were measured before anaesthesia, 2 min after intubation, before incision, 2 min after sternotomy, 10 min after the end of CPB, after chest closure, 30 min and 2h after arrival of the patient in the intensive care unit. The only difference found between the two groups was a more rapid drop in left ventricular preload after induction with sufentanil; 2 min after intubation, there was a 26% fall in Ppw with sufentanil (p less than 0.01) and 8% with fentanyl. Before skin incision, this drop was of 32% (p less than 0.01) and 24% (p less than 0.01) respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975926 TI - [Calcium inhibitors and anesthesia]. AB - Calcium blockers (CB) are routinely used. This could lead to possible interference with anaesthetic drugs. CB prevent calcium from entering the cell by inhibiting the slow voltage-dependent calcium channels. They act mostly on heart and smooth muscle. Of all the possible indications, the three that are confirmed are coronary heart disease, arterial hypertension and supraventricular rhythm disturbances. Most of the work published and the cases reported concerns interactions between CB and halogenated anaesthetic agents; the latter's actions on the heart depend on cellular calcium exchange. Also, the cardiovascular effects of these anaesthetics are similar to that of CB. Experimentally, halothane and enflurane have direct cardiac inhibitory effects similar to verapamil and diltiazem, whereas isoflurane's properties seem closer to the dihydropyridines (nifedipine and nicardipine). Giving verapamil or diltiazem increases the number of sino-atrial and atrio-ventricular blocks when using a halogenated agent. Clinically, interpreting the effects of CB during anaesthetic induction is difficult because of the pathology (coronary heart disease, cardiac failure), the other drugs (beta-blockers and nitrates) and the type of anaesthesia (emergency or elective). Interactions can give rise to anything from a severe cardiovascular collapse, requiring catecholamines, to a mild fall in blood pressure which responds well to plasma expansion, or even no effect on blood pressure. Rebound is seen on stopping CB in patients with coronary heart disease or arterial hypertension; stopping them before surgery does not therefore seem justified. However, extreme care must be taken when using halogenated agents for patients under treatment with CB and/or beta-blockers. A wary anaesthetist will be able to adapt the technique to the patient. It has been suggested that CB could be used to treat preoperatively myocardial ischaemia (diltiazem), hypertensive crises (nifedipine, nicardipine) and ventricular rhythm disturbances (verapamil); this must be done with caution, the patient being closely monitored (haemodynamic and electrocardiographic monitoring). Postoperatively, intranasal nifedipine, continuous intravenous nicardipine or diltiazem have been used to treat increases in arterial blood pressure during recovery and to adapt the cardiovascular system to the increased metabolic needs. Here again, close patient monitoring is essential. In any case, treatment with CB which has been stopped should be started up again as soon as possible. PMID- 2975928 TI - [New antibiotics in 1988]. PMID- 2975927 TI - [The impossible handicap. Analysis of the concept of handicap in the orientating law of 30 June 1975. 1]. AB - This study focuses on handicap as is meant in the French law of june 30, 1975 for handicapped persons. The method is content analysis applied to the law itself and to directly relevant official texts (Bloch Laine report of 1967, motives statement of the law, debates in parliament, Cour des Comptes report of 1982, Lasry Gagneux report of 1983). Handicap thus explicited is twice impossible. In the first place the whole field of handicap is built upon a strong relationship between official services and the invalids while at the same time the law gives the committees total freedom to choose and change their definition of handicap. In the second place the fact of giving such a central place to invalidity and to the relationship between official services and invalids tends to deny the very characters of handicap. P.H.N. Wood's concept as published under the authority of World Health Organization, provides a possible definition of handicap in as much as it states its social nature (to be differentiated from invalidity) and it grants equal importance to the three interactions: person-state, state-milieu, and person-milieu. PMID- 2975929 TI - Experience with the Nd:YAG laser in the treatment of keloid scars. AB - Numerous therapeutic modalities have been espoused for the treatment of keloids. In the laboratory, great strides have been made in understanding the connective tissue pathobiology that underlies this problem. Unfortunately, no consistent treatment has been found to prevent the development of surface overhealing. Moreover, all forms of therapy designed to treat this condition have produced inconsistent results. This article documents the use of the Nd:YAG laser in the treatment of 20 patients with keloid scars. Candidates for the study included patients with no previous therapy and those who had prior treatment with steroids and/or surgery with no improvement. Although the use of Nd:YAG laser for keloids continues to be investigational, our results appear promising. Success of therapy was judged by softening and flattening of the scar. Laboratory studies provide a basis for understanding the possible effect of connective tissue metabolism with the Nd:YAG. PMID- 2975930 TI - [Percutaneous transluminal angioplasty. Is there still a role for the surgeon?]. AB - Relating their own clinical experience and that of the medical literature, the authors examined the role of the surgeon in coronary angioplasty. Despite the considerable progress made in the management of accidents, they believe that one must be able to resort to surgery--even if the percentage of patients who undergo emergency surgery decreases significantly--in the form of a standby procedure organized according to the difficulty of the angioplasty. Moreover, the surgeon uses this technique during the procedure for additional revascularization above the graft on collateral arteries and angioplasty can and should be used in a higher number of cases as a supplement for coronary bypass procedures. PMID- 2975931 TI - [What strategy for treating the acute phase of myocardial infarction?]. AB - Therapeutic strategy of treatment of the acute phase of myocardial infarction has been considerably modified the last few years by the administration of drugs which protect the ischemic myocardium (beta-blockers, calcium-channel inhibitors, notably), or in preventing development of necrosis by reperfusing it either by coronary bypass procedures, or by intracoronary thrombolysis or coronary angioplasty. But these methods are difficult to institute rapidly. Thus, we prefer to perform intravenous thrombolysis, which now may even be given in the patient's home. Clinical experience obtained by the SAMU medical emergency unit in Marseille, France shows the feasibility of such an approach. Initial results are satisfactory. The question of coronary angioplasty as a supplementary procedure may then be discussed: which patients are candidates for this method and when should it be performed? PMID- 2975932 TI - [Atrial natriuretic factor in cardiovascular pathology]. AB - Atrial natriuretic factor (ANF) is a cardiac peptide hormone whose detection led to the discovery of a new natriuretic vasomotor relaxant hormonal system where the heart plays the role of an endocrine gland. Atrial distension represents the main stimulus for the release of ANF. Its cardiovascular effects consist primarily of hypotension related to its relaxant properties, a decrease in cardiac output and a negative inotropic effect. The close relations between ANF and the heart implicate this hormone as a major factor in all cardiovascular disorders, and in particular in congestive heart failure where its plasma concentration represents an index of hemodynamic and functional disease severity. Hemodynamic changes related to valvular heart disease, whether of the mitral or aortic valve, as well as dysrhythmias have a significant effect on release of ANF. Thus, it is now well recognized that ANF, released by the heart, is implicated in the pathophysiology of cardiovascular disorders. PMID- 2975933 TI - [Transluminal coronary angioplasty. Ambitions and difficulties of the second generation]. AB - Proposed in all functional forms of coronary heart disease and by gradually surmounting the obstacles of coronary arteriography, ballon catheter angioplasty, at the same time measures its limitations. These are recurring stenoses and failure of this procedure to cure chronic coronary occlusions, and they specifically characterize a method which, at the expense of unavoidable parietal lesions, does not eliminate the cause of the stenosis. It appears more desirable to remove atheromatous tissue by its extraction, vaporization or pulverization, to reinforce the wall of the coronary vessel, and these are the objectives of second-generation procedures. Development of angioscopy, lasers, atherotomies, and endoprostheses illustrates the vitality and the innovation present in invasive methods of cardiac therapeutic management. However, for the time being their clinical development of such procedures is incomplete and has not been sufficiently perfected. Future methods of angioplasty will be more satisfactory and have better control, but at present it is difficult to accurately predict the future aspects of such procedures. PMID- 2975934 TI - Parapsoriasis and related conditions. AB - Classification of parapsoriases is revised into a simple practical table. Pityriasis lichenoides (guttate parapsoriasis) is not a type of parapsoriasis. The clinical features of small patch and large plaque parapsoriasis are described in detail. Six clinical varieties of large plaque parapsoriasis and three clinical varieties of exfoliative dermatitis including Sezary syndrome have been clearly recognized as distinctive categories. Histopathology is useful for the diagnosis of parapsoriasis and mycosis fungoides. Sezary cell count is not significant for the diagnosis of Sezary syndrome. PMID- 2975935 TI - Fixed drug eruptions--a Singapore study. AB - This study documents 62 patients with fixed drug eruptions (FDE) proven by provocative testing. Of these 39 (63%) were caused by tetracycline hydrochloride and 12 (19%) caused by Saridon (R). Fifty-eight (94%) of the patients had 10 or less lesions and 50 (81%) had 5 or less lesions. The lips and genitalia were preferential sites being involved in 28 (45%) and 22 (36%) instances. In 6 instances each a solitary lesion occurred on the lower lip or the glans penis. At presentation, 43 (69%) patients were unaware of the drug-induced nature of their condition. Compared with previous reports the causative drugs varied from place to place and from decade to decade. This may be due to differences in prescription patterns. PMID- 2975936 TI - HLA and antibiotic allergic skin eruption. AB - Chinese patients with cutaneous eruption due to antibiotics were found to be positively associated with HLA-BW46 (RR = 2.5; 95% confidence limit 1.2-4.9) and negatively associated with HLA-B40 (RR = 0.46; 95% confidence limit 0.22-0.98). The association with HLA-BW46 was even higher in patients with ampicillin sensitivity (RR = 3.5; 95% confidence limit 1.4-9.0). PMID- 2975937 TI - Generalised exfoliative dermatitis--a clinical study of 108 patients. AB - This study was to investigate the epidermiological, etiological, clinical and prognostic aspects of patients with Generalised Exfoliative Dermatitis in Singapore. From 1981 to 1985, all patients with exfoliative dermatitis or erythroderma admitted to our dermatology wards were included into this study. A standard protocol was filled in for each patient, and they were subsequently followed up at our out-patient clinics. There were 79 male and 29 female patients (sex ratio 2.7:1). The average age was 61.5 years. Most of the patients were from low social income groups. Apart from scaling and erythema, peripherial oedema was the most common finding (44%), followed by hair loss (25%) and lymphadenopathy (22%), 15% of patients had haemoglobin level 10G/dl. The main etiological groups were eczema/dermatitis, psoriasis, drug reactions and those with unknown causes. Two patients were suffering from congenital dermatoses. None of our patients was casually related to malignancy. This study suggested that GED was not a fatal disease among our patients, and they often had a good prognosis. PMID- 2975938 TI - Occupational dermatoses--an update. AB - Occupational dermatosis is a common occupational disease. Contact dermatitis is its commonest presentation. New chemicals are introduced into the industry every year. Often they are potential skin irritants and allergens. Besides contact dermatitis, these chemicals can cause serious systemic effects which sometimes produce cutaneous manifestations e.g. chloracne and scleroderma. Prevention remains the most effective way of reducing the incidence of occupational dermatoses. Physicians should be familiar with advances in occupational dermatology in order to recognize them. This paper reviews some recent advances in this field. PMID- 2975939 TI - [Thyroid function and trisomy 21. TSH increase and rT3 deficiency]. AB - An excess of thyrotropin (TSH) with normal levels of tetraiodothyronine (T4) and of 3,5,3'-triiodothyronine (T3) was confirmed in the serum of 78 trisomy 21 children. A severe deficiency of 3,3',5'-triiodo-thyronine (rT3 or reverse T3) was observed and the decrease of the rT3/TSH ratio was highly significant. These new facts suggest that the rT3 deficiency plays a peculiar role in trisomy 21 (maybe through the regulation of one or few steps of monocarbons' metabolism). A systematic control of thyroid function (including the patient's rT3 level) is mandatory for the follow-up of every trisomy 21 patient. PMID- 2975940 TI - A genetic study of the Konda Kapu tribe of coastal Andhra Pradesh, South India. AB - Phenotype and gene frequencies of two blood group and four red cell enzyme systems were examined in a Konda Kapu tribal sample of Coastal Andhra Pradesh, South India. The gene frequencies for these systems in Konda Kapus indicate the middle range values for Andhra Pradesh tribal populations, excepting the ADA and Rh(D) systems, where extreme range values are found. Further, gene flow is indicated between the Konda Kapus under study and Plain Kapus, a neighbouring caste population by calculation of Fi estimates. PMID- 2975942 TI - [Transluminal angioplasty of the coronary vessels in patients with pluritruncular unstable angina]. AB - Between October, 1979 and August, 1987, 489 patients with multivessel coronary disease and unstable angina underwent transluminal angioplasty of coronary arteries with the following results: primary success in 90 p. 100 of the patients, emergency bypass surgery of occlusive dissection in 1.8 p. 100, myocardial infarction in 2.9 p. 100, death in 1.4 p. 100. These results were similar to those obtained in 369 patients with stable angina whose coronary vessels were dilated by the same group during the same period. The death rate was significantly higher in elderly people and in women. 398/489 patients were followed up for 2 to 45 months: 1.8 p. 100 died, 2 p. 100 developed myocardial infarction and 2.3 p. 100 underwent coronary bypass. 46 patients had repeat angioplasty for restenosis. After single or repeat angioplasty, 68 p. 100 of the primary success patients followed up were asymptomatic, and 73 p. 100 had lasting clinical improvement. Among 221 patients studied with different numbers of vessels treated, the degree of revascularization did not make any significant difference in the percentage of symptom-free patients. Data from the literature concerning the medical treatment of unstable angina indicate a high incidence of complications and a mediocre long-term functional benefit, while data concerning surgical treatment show a better long-term functional result. Compared with these two types of treatment, transluminal coronary angioplasty appears as a satisfactory method to treat unstable angina in patients with multivessel coronary disease. PMID- 2975941 TI - Reconstitution of the gastrointestinal microflora of lactobacillus-free mice. AB - A colony of mice that do not harbor lactobacilli in their digestive tracts but whose intestinal microflora is otherwise functionally similar to that of conventional animals was derived. Methods used to reconstitute the intestinal microflora of the mice included inoculation of the animals with cultures of specific microbes, noncultivable microbes attached to epithelial cells, and cecal contents from conventional mice treated with chloramphenicol. Twenty-six microflora-associated characteristics were monitored by using relatively simple tests to determine the microflora status of the mice. PMID- 2975943 TI - Group training to increase social behaviors in young multihandicapped children. PMID- 2975944 TI - ATP-dependent interaction of propranolol and local anaesthetic with sarcoplasmic reticulum. Stimulation of Ca2+ efflux. AB - Preincubation of sarcoplasmic reticulum (SR) with propranolol or tetracaine inhibits Ca2+ accumulation and stimulates ATPase activity by more than 2-fold. This effect is obtained only when the preincubation is carried out in the presence of ATP or other nucleoside triphosphates. The (ATP + drug)-induced inhibition of Ca2+ accumulation is pH-dependent, increasing as the pH rises above 7.5. The presence of micromolar concentrations of Ca2+ or Mg2+ during the preincubation prevents the inhibitory effect of ATP plus drug on Ca2+ accumulation or ATPase activity. The (ATP + drug) modification of SR vesicles resulted in stimulation of a rapid Ca2+ efflux from passively loaded vesicles. The ATP-dependent inhibition of Ca2+ accumulation by the drug is obtained with other local anaesthetics. The drug concentration required for 50% inhibition was 0.15 mM for dibucaine and 0.4 mM for both propranolol and tetracaine, whereas it was 5 mM, 8 mM and greater than 10 mM for lidocaine, benzocaine and procaine respectively. The heavy SR vesicles were only slightly affected by the incubation with propranolol or tetracaine in the presence of ATP, but their sensitivity increased markedly after storage at 0 degrees C for 24-48 h. These results suggest that propranolol and some local anaesthetics, in the presence of ATP, stimulate Ca2+ efflux by modifying a protein factor(s) rather than the phospholipid bilayer. PMID- 2975945 TI - Insulin proteinase liberates from glucagon a fragment known to have enhanced activity against Ca2+ + Mg2+-dependent ATPase. AB - We find, contrary to previous reports, that substantial cleavage of glucagon by insulin proteinase occurs at only one region, namely the double-basic sequence Arg17-Arg18-. Cleavage takes place almost exclusively between these two residues, liberating fragments glucagon-(1-17) and glucagon-(18-29). Others have shown that the fragment glucagon-(19-29) is 1000-fold more efficient compared with intact glucagon, at inhibiting the Ca2+-activated and Mg2+-dependent ATPase activity and the Ca2+ pump of liver plasma membranes. We show that this fragment is not liberated in detectable quantities by our insulin proteinase preparation. On the other hand, others have shown that glucagon-(18-29), though less active than glucagon-(19-29), was still 100-fold more active than glucagon itself in the above-mentioned system. Our observations represent the first demonstration of the release by insulin proteinase of a hormone fragment having enhanced activity, although it has yet to be shown that the activity of this fragment is important in vivo. Since the formation of glucagon-(19-29) from glucagon-(18-29) would involve merely removal of Arg18, a second enzyme might exist to provide the more active fragment. PMID- 2975946 TI - Evidence that a novel serine kinase catalyses phosphorylation of the insulin receptor in an insulin-dependent and tyrosine kinase-dependent manner. AB - Insulin receptor was co-purified from human placenta together with insulin stimulated kinase activity that phosphorylates the insulin receptor on serine residues. By using this 'in vitro' system, the mechanism of activation of the serine kinase by insulin was explored. Peptide 1150, histone, poly(Glu-Tyr), eliminating Mn2+ (Mg2+ only), treatment at 37 degrees C (1 h), N-ethylmaleimide, phosphate, beta-glycerol phosphate and anti-phosphotyrosine antibody all inhibited insulin-receptor tyrosine kinase activity and the ability of insulin to stimulate phosphorylation of the insulin receptor on serine. Additionally, direct stimulation of the receptor tyrosine kinase by vanadate increased serine phosphorylation of the insulin receptor. Insulin-stimulated tyrosine phosphorylation preceded insulin-stimulated serine phosphorylation of the insulin receptor. The activity of the insulin-sensitive receptor serine kinase was not augmented by cyclic AMP, cyclic GMP, Ca2+, Ca2+ + calmodulin, Ca2+ + phosphatidylserine + diolein or spermine, or inhibited appreciably by heparin. Additionally, the serine kinase phosphorylated casein or phosvitin poorly and was active with Mn2+. This indicates that it is distinct from Ca2+, Ca2+/phospholipid, Ca2+/calmodulin, cyclic AMP- and cyclic GMP-dependent protein kinases, casein kinases I and II and insulin-activated ribosomal S6 kinase. Taken together, these data indicate that a novel species of serine kinase catalyses the insulin-dependent phosphorylation of the insulin receptor and that activation of this receptor serine kinase by insulin requires an active insulin-receptor tyrosine kinase. PMID- 2975947 TI - General pharmacology of ramipril. AB - General pharmacological properties of (+)-(1S,3S,5S)-2-[(S)-N-[(S)-1 ethoxycarbonyl-3-phenyl-propyl] alanyl]-2-azabicyclo[3.3.0]octane-3-carboxylic acid (ramipril, Hoe 498), a new prodrug non-sulfhydryl angiotensin converting enzyme inhibitor and its active diacid metabolite, ramiprilat, were examined. Both ramipril and ramiprilat were without effect on basal central and autonomic nervous systems in rats and mice. Ramipril given intravenously to anaesthetized normotensive dogs produced a slight fall in blood pressure but did not significantly alter other cardio-hemodynamic functions. Also, ramiprilat was without effect on isolated atria and airway resistance of guinea pigs. Oral administration of ramipril to dogs increased renal blood flow but did not significantly affect other renal parameters, for example, glomerular filtration rate and electrolyte excretion. Ramipril produced a competitive inhibition of late proximal tubular secretion which points to in part renal secretory excretion of ramipril and/or its metabolites. Compared to urea-induced diuresis in rats, ramipril was without direct diuretic activity. Ramipril exerted little, if any, influence on gastric bile and pancreatic secretion or intestinal transit in rats, as well as on concentration of glucose and lipoproteins, blood coagulation, platelet aggregation and vascular permeability in rats, rabbits or dogs. The carrageenin-induced rat paw edema was enlarged by ramipril, but there was no such effect on serotonin-, dextran- or ovalbumin-induced edemas which in contrast to carrageenin do not involve bradykinin. Thus, undesired cutaneous reactions might result from locally released bradykinin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975948 TI - Dental sedation. A review. PMID- 2975949 TI - Lack of effect of warfarin on the restenosis rate or on clinical outcome after balloon coronary angioplasty. AB - Between September 1985 and April 1987, 110 consecutive patients who had successful coronary angioplasty were included in a randomised prospective controlled evaluation of the effects of warfarin on restenosis. The warfarin (n = 56) and the control (n = 54) groups were not different in terms of age, sex, previous coronary bypass surgery or coronary balloon angioplasty, severity of symptoms, and frequency of multivessel disease or of total coronary occlusions. Warfarin was started on the day of the procedure and the dosage was adjusted to maintain the thromboplastin international normalised ratio greater than or equal to 2.5. One hundred and five (96%) of the patients were given verapamil and other antianginal drugs were prescribed as needed. Low molecular weight dextran and heparin were given during the procedure and heparin was continued for 24 hours in all patients. One hundred and eight (98%) of patients were followed up clinically after a median of five months (range 1-20). Eighty five (77%) had follow up angiography at five months. In the warfarin group symptoms improved in 46 (85%) patients by at least 1 angina class and 31 (57%) were symptom free; the exercise test remained positive in 20 (36%) patients and the angiographic restenosis rate was 25% per lesion and 29% per patient. There were no major bleeding complications. In the control group 46 (85%) patients were improved by at least 1 angina class and 31 (57%) were symptom free; the exercise test was positive in 11 (21%) patients and the angiographic restenosis rate was 33% per lesion and 37% per patient. Although the incidence of angiographic restenosis tended to be lower with warfarin, none of these differences was significant. These data suggest that the combination of verapamil and warfarin, in the absence of aspirin, is not significantly better than verapamil alone in preventing symptom recurrence or angiographic restenosis after coronary angioplasty. PMID- 2975950 TI - Anomalous left coronary artery arising from the pulmonary artery in infancy: is early operation better? AB - From January 1975 to January 1987, 21 consecutive infants aged less than six months (mean (SD) 2.6 (1.2] were admitted with anomalous origin of the left coronary artery from the pulmonary artery. In the first 12 patients, who were seen up to 1982, operation was performed after the age of one year (mean (SD) 29 (29) months) (group 1). The next nine infants, seen from 1983 to 1987, had their operations within a few weeks of the onset of symptoms (mean (SD) age 4.8 (1.4) months) (group 2). In group 2 the left coronary artery was relocated into the aorta, whereas in group 1 there was additional resection of the left ventricular wall or mitral valvoplasty or both. At presentation there were no differences in age, clinical condition, heart enlargement, and echocardiographic left ventricular dysfunction between groups 1 and 2. Seven of the 12 patients in group 1 died, five while they were awaiting operation (three died suddenly at home) and two at operation. The five survivors are doing well 6.4 (3.1) years after operation with normal left ventricular function which improved slowly over several months after operation. Two of the nine patients in group 2 died; both deaths occurred at or soon after operation. The seven survivors are doing well 1.8 (0.9) years after operation. In three, left ventricular function recovered within three weeks; and there was even partial or total regression of the Q waves in the supposedly necrotic areas. In the remaining four the pattern of improvement in left ventricular function resembled that in group 1. Operation should be undertaken early in infants with anomalous left coronary artery arising from the pulmonary artery because the procedure is relatively safe, prevents a high natural mortality, and offers a better chance of a faster recovery of left ventricular function. PMID- 2975951 TI - Survival, response and immune effects in a prospectively randomized study of dose strategy for alpha-N1 interferon. AB - Several tumour sites have now demonstrated objective responses to alpha interferons in a diversity of doses and schedules. Since effectiveness should be enhanced with the identification of an optimal dose strategy, we undertook a prospectively randomized study to compare an intermittent high dose escalating strategy (HDS) vs. a fixed low dose treatment in relation to clinical outcome and laboratory correlates of immune function. HDS patients received interferon alpha N1 (lymphoblastoid interferon) 5M units m-2 by continuous i.v. infusion over 24 h, escalating by 5 M units m-2 day-1 as tolerated over 10 days, and repeated every 28 days. The low dose strategy (LDS) consisted of a fixed dose of 2 M units m-2 by intramuscular injection daily for 28 days, then daily for 7 days every other week. There were 53 evaluable patients. In keeping with earlier preliminary results there was evidence of improved immune function for HDS patients. They demonstrated a significant increase in the number of CD2+ (sheep red blood cell binding) cells and CD4+ (helper-inducer/suppressor-inducer) cells along with enhanced activity of natural killer cell, and mixed leukocyte culture activity. In addition to improved immune function, HDS patients survived longer than LDS (P = 0.04). Analysis of survival in relation to response suggested that monitoring of minor responses may be of interest for biological agents such as interferon. PMID- 2975952 TI - Immunohistological localization of pregnancy-associated endometrial alpha 2 globulin (alpha 2-PEG) in endometrial adenocarcinoma and effect of medroxyprogesterone acetate. AB - Endometrium from postmenopausal women with endometrial adenocarcinoma was examined immunohistochemically using a monoclonal antibody to pregnancy associated endometrial alpha 2-globulin (alpha 2-PEG), the major secretory protein of the glandular epithelium during the late luteal phase of the menstrual cycle and early pregnancy. Specimens were obtained at initial diagnostic curettage and at hysterectomy after medroxyprogesterone acetate (MPA) therapy. alpha 2-PEG was not detected in any malignant tissue irrespective of histological differentiation. Non-malignant endometrium obtained in association with malignant tissue was negative for alpha 2-PEG before treatment although after MPA therapy all specimens obtained exhibited marked alpha 2-PEG localization in glands. In four specimens endogenous alkaline phosphatase was observed consistently only in the malignant endometrium. Malignant endometrium does not appear to synthesize alpha 2-PEG nor is its synthesis induced by an oral progestogen, so that it does not represent a useful marker for endometrial carcinoma. Non-malignant endometrium in postmenopausal women appears to be fully capable of alpha 2-PEG production after stimulation with an oral progestogen. PMID- 2975953 TI - The binding of epidermal growth factor to the human uterus and leiomyomata in women rendered hypo-oestrogenic by continuous administration of an LHRH agonist. AB - The binding of epidermal growth factor (EGF) to human myometrium and leiomyomata was assessed in a group of women rendered hypo-oestrogenic with the LHRH agonist Zoladex (ICI 118630). The results were compared with those obtained with tissues from women with normal cycles. In normal women, the specific binding of radiolabelled [125I] EGF to both myometrial and fibroid homogenates did not vary during the menstrual cycle, but the specific binding of [125I] EGF to fibroid in women treated with LHRH agonist was significantly less than in the untreated group. Since the hypo-oestrogenic state induced by the agonist is associated with a decrease in fibroid size, the results suggest that the effect of oestrogen on fibroid tissue may partly be mediated by EGF. PMID- 2975954 TI - Evaluation of dye penetrations around two resin-based materials used as fissure sealants. AB - Two materials, based on Bis GMA resin, which may be used as fissure sealants were evaluated for dye penetration between sealant and enamel. The conventional fissure sealant (Durafil Flow) showed the greatest degree of dye penetration in the largest number of specimens. The penetration of the dye was less extensive when a filled resin (P30) was used. The use of a dentine adhesive together with the filled resin further reduced the penetration of the dye. The number of teeth exhibiting leakage with this treatment was the smallest noted. PMID- 2975955 TI - Moderate resolution profile structure of the sarcoplasmic reticulum membrane under low temperature conditions for the transient trapping of E1 approximately P. AB - The calcium uptake reaction kinetics of isolated sarcoplasmic reticulum (SR) vesicles have previously been shown to be at least biphasic over a range of temperatures (26 to 10 degrees C) with a fast phase identified with the formation of E1 approximately P and calcium occlusion and a slow phase with Ca2+ translocation across the membrane and turnover of the Ca2+ ATPase ensemble. At "low" temperatures, namely 0 degrees C or lower, E1 approximately P formation is slowed and E1 approximately P is transiently trapped for at least several seconds, as indicated by the absence of the slow phase for 6 s or more. We now report that a reversible, temperature-induced structural transition occurs at about 2-3 degrees C for the isolated SR membrane. We have investigated the nature of this structural transition utilizing meridional and equatorial x-ray diffraction studies of the oriented SR membrane multilayers in the range of temperatures between 7.5 and -2 degrees C. The phase meridional (lamellar) diffraction has provided the profile structure for the SR membrane at the highest vs. lowest temperature at the same moderate resolution of 16-17 A while the equatorial diffraction has provided information on the average lipid chain packing in the SR membrane plane in the two cases. To identify the contribution of each membrane component in producing the differences between the profile structures at 7.5 and -2 degrees C, step-function models have been fitted to the moderate resolution electron density profiles. Lipid lateral phase separation may be responsible for inducing the structural change in the Ca2+ ATPase, thereby resulting in the slowing of E1 approximately P formation and the transient trapping of E1 approximately P at the "lower" temperatures. PMID- 2975957 TI - Therapy of advanced ovarian cancer with D-Trp-6-LH-RH (decapeptyl) microcapsules. AB - Forty-one patients with advanced ovarian cancer (FIGO stage III or IV) who had relapsed following conventional treatment were treated with long acting depot preparation of D-Trp-6-LH-RH once a month. There were no exclusion criteria and patients of any age or performance status were eligible. Thirty patients continued to progress following therapy. Twenty-six patients died within the first 16 weeks from onset of therapy. Five patients remained stable (SD) on D-Trp 6-LH-RH with no change in tumor size. Six patients had evidence of clinical and/or radiological partial remission (PR) where the tumor size decreased by more than 50%. The remissions in the responding patients were maintained for a mean duration of 10 months (range 6-18 months). Thus 11 patients (6 PR + 5 SD) have had remission or stabilization of disease which has been clinically worthwhile. Clinical benefit of therapy with microcapsules of slow-release D-Trp-6-LH-RH in about 26% of patients is highly encouraging. Treatment with D-Trp-6-LH-RH offers an important non-toxic alternative in patients who do not tolerate chemotherapy or who have progressive disease following chemotherapy. PMID- 2975956 TI - Changes in the sarcoplasmic reticulum membrane profile induced by enzyme phosphorylation to E1 approximately P at 16 A resolution via time-resolved x-ray diffraction. AB - Time-resolved x-ray diffraction studies of the isolated sarcoplasmic reticulum (SR) membrane have provided the difference electron density profile for the SR membrane for which the Ca2+ ATPase is transiently trapped exclusively in the first phosphorylated intermediate state, E1 approximately P, in absence of detectable enzyme turnover vs. that before ATP-initiated phosphorylation of the enzyme. These diffraction studies, which utilized the flash-photolysis of caged ATP, were performed at temperatures between 0 and -2 degrees C and with a time resolution of 2-5 s. Analogous time-resolved x-ray diffraction studies of the SR membrane at 7-8 degrees C with a time resolution of 0.2-0.5 s have previously provided the difference electron density profile for the SR membrane for which the Ca2+ ATPase is only predominately in the first phosphorylated intermediate state under conditions of enzyme turnover vs. that before enzyme phosphorylation. The two difference profiles, compared at the same low resolution (approximately 40 A), are qualitatively similar but nevertheless contain some distinctly different features and have therefore been analyzed via a step-function model analysis. This analysis was based on the refined step-function models for the two different electron density profiles obtained independently from x-ray diffraction studies at higher resolution (16-17 A) of the SR membrane before enzyme phosphorylation at 7.5 and -2 degrees C. The step-function model analysis indicated that the low resolution difference profiles derived from both time resolved x-ray diffraction experiments arise from a net movement of Ca2+ ATPase protein mass from the outer monolayer to the inner monolayer of the SR membrane lipid bilayer. The conserved redistribution of this protein mass is however somewhat different for the two cases, especially at the extravesicular membrane surface containing the Ca2+ATPase "headpiece." However, the conserved redistribution of protein mass within the SR membrane lipid bilayer common to both cases is clearly due to E1~P formation. PMID- 2975958 TI - Two drug-induced dermatological syndromes among employees of a Polish construction site in a tropical country. AB - The two rare cases of exudative multiform erythema were described in its heavier alterations of Fiessinger-Rendu-Leroy as well as of Stevens-Johnson syndrome, most likely of drug-induced origin, at the employees of a Polish construction site in a tropical country, taking under consideration the causing reasons, diagnostic features as well as conditions of course and principles of treatment. PMID- 2975959 TI - Screening for Down's syndrome. PMID- 2975960 TI - The disabled living centre: what does it do? AB - Seventy five consecutive users of the Leeds disabled living foundation were surveyed. Two thirds of these users were at least moderately severely disabled. Forty eight of the 65 people (74%) who completed the questionnaire were recommended aids, and 33 had received some at the end of the survey. Only 10 of 28 people had received recommended adaptations. Few referrals were made to the centre by doctors. It is important that they tell their patients of this useful service. PMID- 2975961 TI - Hypothyroidism in Down's syndrome. AB - Fifty-five adult Down's syndrome subjects resident at Northgate Hospital were screened for the presence of thyroid dysfunction. Approximately 50% of the subjects had clinical features suggestive of hypothyroidism, sufficient to require thyroid-function tests. Twenty-two per cent of the total suffered from some degree of hypothyroidism, and 16% had positive antibodies to thyroid tissue. PMID- 2975962 TI - Comparison of the biopotencies of nicotinic acid and nicotinamide for broiler chicks. AB - 1. Two experiments were conducted with 1056 1-d-old Cobb x Cobb feather-sexed broiler chicks, to determine the relative nicotinic acid biopotencies of nicotinic acid (NA) and nicotinamide (NAm). 2. Two methods were used: the slope ratio technique and the broken-line model. 3. In experiment 1, using the slope ratio technique, NAm was 0.82 as active as NA in promoting growth to 21 d of age. In experiment 2 NAm was 0.96 as bipotent as NA. 4. Applying the broken-line model to the body weight data at 21 d of age for both NA and NAm in experiment 1, it was found that NAm had approximately 0.82 NA activity. The same analysis applied to the data of experiment 2 indicated that the biopotency of NAm was 1.09. 5. A t test of the slope-ratio values obtained in both experiments indicated that the average (0.89) was not significantly different from 1.0. 6. It was concluded that the relative biopotencies of NA and NAm in broiler chicks fed a maize-soyabean meal diet were similar. PMID- 2975963 TI - Streptozotocin toxicity to cultured pancreatic islets of the Syrian hamster. AB - Pancreatic islets of the Syrian golden hamster were maintained in culture for extended periods of time. Toxicity of streptozotocin in these cultures was evaluated by measurement of insulin secretion. Exposure of islets to 1 or 2 mM streptozotocin immediately following isolation resulted in a permanent and dose related inhibition of insulin secretion. This was accompanied by islet disruption as observed by phase-contrast microscopy. Culture of islets for 24 hours before streptozotocin exposure afforded protection from toxicity. For example, exposure of freshly isolated islets to 2 mM streptozotocin resulted in complete destruction of beta cells, whereas islets similarly exposed after a 24 hr culture period continued to secrete insulin for many months. Islets maintained in culture for one week before exposure to 0.1-0.5 mM streptozotocin, however, became more sensitive than freshly isolated islets. Repeated weekly exposure of cultured islets to a "non-toxic" concentration (0.1 mM) resulted in sustained suppression of insulin secretion after 11 weeks. PMID- 2975964 TI - Trimetrexate: molecular structures and conformational similarities in two crystal forms. AB - The structure of the non-classical quinazoline antifolate trimetrexate (TMQ) has been determined in two crystal forms, TMQ acetate monohydrate, and hydrated TMQ free base. Trimetrexate has an extended conformation in both structures, and the quinazoline and phenyl rings are mutually perpendicular. Protonation occurs at N1 in the acetate salt. The TMQ conformation is similar to corresponding parts of quinespar, the only other quinazoline antifolate structurally determined, and the hydrated strontium salt of methotrexate. PMID- 2975965 TI - Diagnostic accuracy of Doppler ultrasound technique of the penile arteries in correlation to selective arteriography. AB - In 63% of 265 patients with erectile dysfunction a relevant arterial inflow disturbance was found by Doppler ultrasound examination. Correlation between Doppler and arteriography in 58 patients showed an accuracy of 95% in detecting penile arteries and an accuracy of 91% in discovering a pathological arterial pattern (arterial anomaly or arteriosclerotic obstruction). In 15 patients the arterial inflow was measured additionally by Doppler ultrasound technique after intracavernosal injection of vasoactive drugs (IIVD) (7.5 mg papaverine and 0.25 mg phentolamine). This technique proved to be more reliable than in the flaccid state and markedly facilitated localization and assessment of pathological changes of the cavernosal arteries. PMID- 2975966 TI - Transluminal angioplasty in the treatment of arteriogenic impotence. AB - Factors bearing on the role of transluminal angioplasty in the management of arteriogenic impotence are considered. Our clinical experience indicates that arteriogenic impotence is frequent, either alone or combined with venogenic impotence. High quality diagnostic angiographic studies and their accurate interpretation are the prime requirements for proper patient selection. Numerous arteriographic adjuncts are required: vasodilation with intracavernosal injection of a papaverine-phentolamine mixture, selective internal pudendal injections, direct magnification, nonionic contrast agents, and tailored radiographic projections. Venogenic impotence must be excluded by cavernosometry and cavernosography. In impotent patients with bilateral leg and hip claudication, dilatation of common or internal iliac stenoses should benefit many cases with pure arteriogenic impotence. In the absence of claudication, angioplasty will be most frequently indicated for distal internal pudendal lesions, using 2-3 mm balloon-catheter systems. Stenoses of intrapenile branches, while common, must await further technological developments before they too may become amenable to transluminal recanalization. Unilateral transluminal angioplasty, when technically successful, should prove clinically successful when patients have been properly selected. Transluminal angioplasty can reduce the cost and morbidity of penile revascularization and may assume a modest role in the treatment of arteriogenic impotence. PMID- 2975967 TI - Histamine modulates local inhibition in the rat hippocampal slice. AB - 1. These experiments investigated the action of histamine on local inhibition in the CA1 region of the in vitro hippocampal slice preparation using a paired-pulse paradigm. 2. We observed that histamine produced a concentration-dependent and reversible attenuation of paired-pulse inhibition. This effect was reduced by the H2 receptor antagonist, cimetidine, and mimicked by the H2 receptor agonist, impromidine. 3. We also observed that histamine produced concentration-dependent effects on the amplitude of the population spike that could be correlated with alterations in the field excitatory postsynaptic potential (EPSP) amplitude and input fiber volley. High concentrations of histamine produced a reduction in the amplitude of the population spike which was always accompanied by a reduction in the EPSP and fiber volley amplitude. 4. These results suggest that histamine, through the occupancy of H2 receptors, acts to modulate the efficacy of the local synaptic circuitry which is involved in producing paired-pulse inhibition in the hippocampus. PMID- 2975968 TI - Presence of ATPase and alkaline phosphatase activities in the starfish sperm acrosome. AB - ATPase activity was cytochemically detected in the peripheral acrosomal component of ionophore-reacted sperm, while alkaline phosphatase activity was demonstrated in the upper and central components of the acrosome and, at fertilization, at the site of sperm-oocyte binding. Supernatants of ionophore treated sperm suspensions were assayed for ATPase, alkaline and acid phosphatase activities. Results suggest that alkaline phosphatase may be involved both in the acrosomal reaction and oocyte jelly lysis but the function of the acrosomal ATPase remains unknown. PMID- 2975969 TI - Usnic acid revisited, its activity on oral flora. AB - The antibacterial activity of usnic acid, the most widely distributed antibiotic among the numerous ones produced by many lichen species has been re-examined and particular attention has been devoted to the activity of optically active forms of usnic acid against Streptococcus mutans. The D(+) enantiomer was found to be more active than the L(+) form and was observed to exert a rather selective activity against S. mutans. Trials carried out in volunteers showed that mouth rinse with D(+) usnic acid preparations exerted a selective and long lasting action against S. mutans, without substantially altering the equilibrium of normal oral bacterial flora. The adherence of S. mutans to smooth surfaces is not increased by the presence of subinhibiting concentrations of D(+) usnic acid. This is at variance with what has been observed with other antibiotics. These characteristics make D(+) usnic acid a suitable candidate for topical use in oral medicine. PMID- 2975970 TI - Blood pressure and metabolic effects of streptozotocin in Wistar-Kyoto and spontaneously hypertensive rats. AB - There are conflicting reports regarding the action of streptozotocin (STZ) on blood pressure (BP) in rats. This study investigated the BP, metabolic and hormonal effects of increasing doses of STZ in Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR), with consideration to methodological aspects. Indirect tail-cuff systolic BP measured in a conscious state was mildly elevated after 2 to 4 weeks and remained so in severely diabetic, emaciated WKY, whereas there were no changes in the SHR. Four and 20 weeks after STZ administration, systolic, mean and diastolic BPs measured in a conscious state with an arterial catheter were unchanged in the diabetic WKY and were decreased in the diabetic SHR. Thus, the changes in BP depended on the method used. Dose dependent increases in blood glucose were similarly evident under conscious and ether-anesthetized conditions. Triglycerides were increased, and blood insulin and thyroxine levels were decreased in both strains. Between-strain comparisons revealed that the hypoinsulinemic response was similar, but the hyperglycemic and hypertriglyceridemic responses were greater in the SHR. The findings provide a data base for further investigation on STZ diabetes. In addition, the results suggest a different BP and metabolic susceptivity to STZ treatment in the SHR and WKY. PMID- 2975972 TI - Possible antagonistic relationship between atrial natriuretic peptide (ANP)-brain natriuretic peptide (BNP) system and brain renin-angiotensin system in central control of body fluid and cardiovascular function. PMID- 2975971 TI - Local inhibition of angiotensin II formation and bradykinin degradation in isolated hearts. AB - The interaction of the converting enzyme (CE)-inhibitor ramipril and the bradykinin (BK)-antagonist D-Arg-[Hyp2, Thi5,8, D-Phe7]BK with angiotensin I (ANG), ANG II and BK were studied in isolated hearts of rats and guinea pigs. In isolated working rat hearts perfusion with ANG I and ANG II reduced cardiac function and coronary flow, increased the activities of lactate dehydrogenase (LDH) and creatine kinase (CK) in the perfusate, decreased high-energy rich phosphates and glycogen in the myocardium and increased duration and incidence of post-ischemic reperfusion arrhythmias. BK on the other hand reduced LDH and CK activities, improved metabolic parameters in the myocardium and reduced reperfusion arrhythmias. In isolated rat hearts pretreatment with ramipril protected against reperfusion arrhythmias and reduced enzyme activities of LDH and CK in the coronary effluent. Cardiodynamic parameters and coronary flow improved and myocardial tissue levels of glycogen, ATP and creatine phosphate (CP) were elevated. Almost identical changes were seen during perfusion with BK. The cardioprotective effects produced by both, the CE-inhibitor and BK, were completely abolished when the BK-antagonist was added to the perfusate, while a smaller inhibition was obtained by indomethacin perfusion. In isolated guinea pig hearts BK increased coronary flow. Single-dose oral pretreatment with ramipril potentiated, whereas perfusion with the BK-antagonist abolished this effect. These data add support to the hypothesis that local inhibition of CE = kininase II contributes to the beneficial effects of CE-inhibitors in the heart. PMID- 2975973 TI - Modulation of Fc and C3b receptor expression on guinea-pig macrophages by lymphokines. AB - The decrease in Fc-receptor-positive cells that occurred during a 6 h incubation of resident and elicited guinea-pig macrophages was partly abrogated when lymphokines were present in the culture. When the same lymphokine preparations were tested on C3b receptor-expression they preferentially sustained the percentage of C3b rosettes formed by resident rather than elicited macrophages. This lymphokine-induced maintenance of Fc and C3b rosettes by cultured macrophages may have been due to an inhibition of receptor release or an increase in receptor synthesis. Supernatants from cultured macrophages contain shed Fc and C3b receptors which inhibit rosette formation by other macrophages. From the demonstration that culture supernatants from both lymphokine-treated and untreated macrophages significantly inhibited Fc and C3b rosette formation by freshly obtained macrophages it seems that the shedding of Fc and C3b receptors is not modified by lymphokines. The maintenance of Fc and C3b rosettes by lymphokines was inhibited by treatment of the macrophages with cycloheximide, suggesting that the lymphokine effect was due to an increase in synthesis de novo of the Fc and C3b receptors. The lymphokine-inducing antigens, BGG and PPD, and control lymphokine preparations were devoid of receptor modifying activity. The reduction in the percentage of Fc rosettes after 6 h culture appears to be due to a loss of Fc receptors for IgG1. Although lymphokines partly inhibited this effect they could not prevent the loss of these receptors following 24 h culture, unlike their action in augmenting the expression of Fc receptors for IgG2. These findings suggest that a selective enhancement of Fc receptor synthesis by lymphokines may modify the functional activities of macrophages. PMID- 2975974 TI - Serum suppressive activity of HIV seropositive patients. AB - The mechanisms by which HIV induces immunosuppression are still poorly understood so far. Several pathways of CD4 cell destruction are known, including cytolysis with or without syncitium formation and killing by cytotoxic effectors of HIV infected or non-infected CD4 cells. However, a discrepancy exists between the small number of actually infected cells in vivo and the extent of HIV-related immunodeficiency. Among other possible immunosuppressive factors, serum blocking factors have been reported, but only in AIDS-related opportunistic infections (OI), i.e. in a quite specific type of full-blown HIV disease. The purpose of this work was to determine whether serum blocking activity was unique to this group of patients, or if it was also expressed in other clinical presentations and, moreover, at earlier stages of the disease. We also attempted to delineate the nature of these seric factors. In order to do so, we assessed serum suppressive activity of 50 HIV seropositive patients, seven with OI, eight with Kaposi's sarcoma (KS), and 35 with no clinical AIDS. Our results confirm the existence of serum inhibiting factors in AIDS, and demonstrate their presence at earlier stages of the disease. They also highlight the fact that the level of serum suppression does not correlate with patients clinical status, but increases with the severity of the disease. The lower the CD4 count, the higher the suppression exerted. Furthermore, we showed that the suppression was at least partly mediated by small size molecules, which are not complement-mediated or directly lymphocytotoxic. On the other hand, this activity does not correlate with the serum level of p24 HIV core protein. The possible relation with other viral components is discussed. The relevance of these data to prognosis and pathogenesis of HIV disease deserves further investigation. PMID- 2975975 TI - T cell distribution is different in follicle-associated epithelium of human Peyer's patches and villous epithelium. AB - Immunohistochemical analyses performed on specimens of normal human ileum showed a significantly raised number of T cells in the follicle-associated epithelium (FAE) of Peyer's patches compared with the epithelium of distant villi. The T cells tended to be clustered in all layers of the FAE and were significantly more numerous adjacent to interruptions of the brush border (revealed by lack of staining for alkaline phosphatase). Such interruptions were taken to indicate 'membrane' (M) cells. Our findings therefore suggested a spatial relationship between M cells and the aggregation of T cells. The ratio of CD4+ to CD8+ T cells (approximately 4:10) was significantly higher in the FAE than in the villous epithelium (approximately 0.6:10). This suggested that the FAE may be involved to a greater extent in induction of 'helper' T cell functions, perhaps depending on luminal antigens transported by M cells, whereas the villous epithelium may be more involved in stimulation of 'suppressor' T cell functions as indicated by recent studies in vitro. PMID- 2975976 TI - Follow-up of prematurely born infants. AB - Long-term developmental outcome of the prematurely born is generally related to birth weight as a reflection of gestational age: The more immature the infant, the greater the risk of abnormal developmental outcome. Due to improvements in fetal and neonatal care over the past 25 years, developmental outcome of the preterm group has steadily improved, now approaching the expected outcome for term neonates for those born weighing over 1,000 g. For the group of very immature infants (birth weight less than 1,000 g) abnormal developmental outcome remains a significant risk. For any individual preterm infant, long-term developmental outcome is generally related to the severity and duration of initial illness and the postdischarge environment. Specific causation of developmental abnormality in a particular infant is usually speculative. The overall improvement in the long-term outcome for preterm infants has been gained at great cost in medical resources and is accompanied by emotional costs to families that remain unmeasured. The key to further reducing the risk of abnormal developmental outcome remains the prevention of prematurity. Until that can be accomplished, continued meticulous attention to all the details of superb fetal and neonatal intensive care must be exercised to minimize the risk of handicap in this vulnerable group of patients. PMID- 2975977 TI - Incisions. PMID- 2975978 TI - Dehiscence, evisceration, and other complications. PMID- 2975980 TI - Interventional cardiac procedures in neonates and infants: state of the art. AB - The currently available interventional cardiac procedures in neonates and infants are at various stages of development. We currently dilate neonates and infants with critical valvular pulmonary and aortic stenosis and postoperative aortic obstruction. We do not routinely dilate native coarctation of the aorta because of the possibility of aneurysm formation, unless the neonate is very sick and acidotic and an operative approach is considered to be high risk. Balloon and blade atrial septostomy are done routinely whenever indicated with a low incidence of morbidity. Coil embolization, endomyocardial biopsy, foreign body retrieval and percutaneous pericardial drainage are relatively safe, and with the currently available instruments these techniques can be performed safely in neonates and infants with the same indications as for older patients. We currently consider stenotic pulmonary veins to be an undilatable lesion and an optimal therapy remains to be defined. Transcatheter closure of PDA and intracardiac shunts is presently limited to older patients, due to the large size of the delivery system devices and cannot currently be used in neonates. Dilation of the pulmonary valve in cyanotic congenital heart disease appears useful, but further experience is needed. PMID- 2975979 TI - Maturation of the heart. AB - The transition from fetal to newborn to adult previously has been associated with a progressive increase in myocardial contractile function. The purpose of this review is to summarize new information on the biochemical and molecular processes that regulate the perinatal maturation of heart function. The developing heart experiences a perinatal increase in the cellular quantity of the sarcoplasmic reticulum and the myofibrils, the organelles that regulate and utilize cytosolic calcium to produce cardiac contraction. It also has become evident that the function of the available ion pump proteins is reduced in fetuses and newborns compared to adults of the same species. The resultant limited regulation of cytosolic calcium concentrations by the sarcoplasmic reticulum enhances the role of the sarcolemma in this process. Although it appears that the perinatal maturation of sarcoplasmic reticular function is not under direct molecular regulation, alternative splicing may regulate its gene products. A great deal of the perinatal maturation of myofibrillar protein function appears to be regulated by molecular processes. This has been best demonstrated in detail for myosin. In some species, such as the rat, much of the perinatal increase in myocardial contractile function can be explained by a parallel change in myosin isoforms. This isoform shift alters the activity of its ion pump, thereby allowing it to utilize more calcium. In contrast, the perinatal maturation of contractile function in several larger species cannot be explained fully by the extent of the molecular changes that have been identified currently in these species. PMID- 2975981 TI - The transverse abdominal island flap for breast reconstruction. A 7-year experience. AB - Reported herein is a 7-year experience with autogenous tissue breast reconstruction. Current refinements in the use of the transverse abdominal island flap are detailed. In addition, there is discussion of guidelines in patient selection, perioperative management, and occurrence and prevention of complications. PMID- 2975982 TI - PL94-142--its potential problems for primary care physicians. PMID- 2975983 TI - [Diagnosis and therapy of irritable colon syndrome]. PMID- 2975984 TI - [Use of drugs in renal insufficiency. Computer program for microcomputer]. PMID- 2975985 TI - [Esophageal diverticula: the role of esophageal motor dyskinesia and gastroesophageal reflux]. PMID- 2975986 TI - [Surgical therapy in vascular diseases and in the acute stage of infectious endocarditis in elderly patients]. PMID- 2975987 TI - [Use of external arteriovenous shunt as access for immediate hemodialysis in chronic uremic patients and successive conversion to internal arteriovenous fistula]. PMID- 2975988 TI - [Drug surveillance of salcatonin: preliminary results on efficacy and tolerance]. PMID- 2975989 TI - [Hyperparathyroidism in the elderly: a rare disease, often unknown]. PMID- 2975990 TI - [Diet therapy of diabetes mellitus]. PMID- 2975991 TI - [Evaluation of pains of sacral region and upper extremities in stomatologists]. PMID- 2975992 TI - Erythrocyte phosphofructokinase and glucose-6-phosphate dehydrogenase in thyroid disorders. PMID- 2975993 TI - Hormonal receptors in endometrial neoplasias. AB - The Authors studied the concentration of cytosolic and nuclear receptor for Estradiol and Progesterone in endometrial hyperplasia and carcinoma, compared with a control group. Comparative study of hormone receptor concentrations in different populations shows: 1) A significant increase in Estradiol receptors in endometrial hyperplasia (p less than 0.01); 2) A decreasing concentration of estradiol receptors with the decreasing of cellular differentiation in endometrial carcinoma (p less than 0.01); 3) A similar tendency and significance for Progesterone receptors; 4) For both receptors the tendency in the nuclear compartment is similar to that in cytosol but the significance is smaller (p less than 0.05). PMID- 2975994 TI - Atrial natriuretic factor enhances vasopressin-induced bradycardia in normotensive (WKY) but not in spontaneously hypertensive (SHR) rats. AB - Effect of elevation of blood atrial natriuretic peptide (rat 1-28 ANF) level on pressor responses and bradycardia elicited by vasopressin (AVP) was compared in conscious normotensive (WKY) and spontaneously hypertensive (SHR) rats 14-16 weeks old, instrumented with arterial and venous catheters 1-2 days prior to the experiment. Blood pressure and heart rate were determined before and after i.v. injection of 2.5, 5.0 and 10.0 ng of AVP under control conditions (infusion of saline) and during infusion of ANF at a rate of 0.3 micrograms kg-1 min-1. Administration of ANF did not influence pressor responsiveness to AVP (determined as a maximum increase of systolic blood pressure) neither in WKY nor in SHR, however, it elongated the pressor response to the lowest dose of AVP in WKY. Atrial natriuretic factor significantly enhanced bradycardic responsiveness to AVP (determined as a maximum decrease of HR) in WKY but not in SHR. Reduction of heart rate in relation to the corresponding elevation of the systolic pressure (delta HR/delta SP) was also significantly greater in WKY but not in SHR. This effect was significant when blood pressure increases elicited by AVP did not exceed 40 mmHg. The results suggest that ANF potentiates reflex bradycardia elicited by AVP. This action may be a mechanism buffering blood pressure increases in response to AVP. Absence of this effect in SHR may be related to the impairment of the reflex control of blood pressure in this strain. PMID- 2975995 TI - Arginine vasopressin dissociates the diuresis and natriuresis due to atrial natriuretic factor in man. AB - The possible interaction between arginine vasopressin (AVP) and atrial natriuretic factor (ANF) in the control of urinary sodium and water excretion was investigated in man. Nine healthy male volunteers undergoing stable maximal water diuresis were studied on four separate occasions. Atrial natriuretic factor 15 pmol kg-1 min-1 or placebo (P) was concomitantly administered against a background infusion of either AVP 0.003 pmol kg-1 min-1 or P; thus the combinations P + P, AVP + P, P + ANF and AVP + ANF were studied. Atrial natriuretic factor caused a significant increase in sodium excretion (UNaV) [+56%], urinary flow rate (V) [+17%] and free water clearance (CH2O) [+23%]; creatinine clearance (Ccr) did not change. Arginine vasopressin reduced V (-58%) and CH2O (-68%) but did not alter UNaV or Ccr. On the AVP + ANF study day, UNaV increased (+64%) as with P + ANF, but V (-44%) and CH2O (-52%) continued to decrease below baseline levels; analysis of variance showed this antidiuresis reflected the prevalent effect of AVP rather than any specific interaction. These results show that AVP is able to dissociate the natriuretic and diuretic effects of ANF. PMID- 2975996 TI - Repeated stimulation of D-1 dopamine receptors increases the circling response to bromocriptine in rats with a 6-OHDA lesion. AB - We have studied the effect of repeated injections of a specific D-1 agonist, (SKF 38393), on circling behaviour in response to a specific D-2 agonist (bromocriptine) and on striatal D-2 DA receptors in rats with a unilateral lesion of the nigrostriatal pathway. Two groups of rats were formed on the basis of their circling response to a first injection of SKF 38393: clear contralateral circling (SKF-positive) and no (or ipsiversive) circling (SKF-negative). A few days later, these rats were tested once for bromocriptine-induced circling. A repeated treatment of 8 injections of SKF 38393 over 4 days was started the next day. The animals were tested again for bromocriptine-induced circling at the end of the repeated treatment with the D-1 agonist. A third group of rats received only the 2 test injections of bromocriptine and a fourth group included intact control animals. During the repeated treatment, the D-1 agonist induced an increase of the circling response up to the fourth injection and a decrease thereafter in the SKF-positive group. This was followed by a pronounced increase of circling in response to the second injection of bromocriptine. This increased sensitivity appears to be due to the chronic treatment with SKF 38393 since animals that did not respond initially to the D-1 agonist (SKF-negative group) and those that received only 2 injections of bromocriptine showed no increase of their circling response. The treatments did not increase the affinity of [3H]spiperone binding for the striatal D-2 dopamine receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2975997 TI - [Histochemical and electron microscopic studies of the trabecular meshwork in primary open-angle glaucoma]. PMID- 2975998 TI - [Histochemical and electron microscopic studies of the trabecular meshwork in normal human eyes]. PMID- 2975999 TI - Immunopathology of uveitis. PMID- 2976000 TI - Randomized double-blind multicenter study comparing acitretin-PUVA, etretinate PUVA and placebo-PUVA in the treatment of severe psoriasis. AB - A randomized double-blind study was designed with 65 patients in order to clarify two points: (1) does addition of a retinoid to psoralen-ultra violet A photochemotherapy (PUVA) of severe psoriasis decrease the UVA energy required to achieve remission, and (2) is there a difference between two retinoids, i.e. etretinate and acitretin. Acitretin-PUVA treatment was significantly superior to placebo-PUVA with respect to several items (decrease in lesional scores after 6 weeks of therapy, number of PUVA exposures, and total dose of UVA until remission). There were also differences between the etretinate-PUVA and placebo PUVA groups, but only the decrease in lesional scores reached statistical significance. PMID- 2976001 TI - Cutaneous metastasis from papillary carcinoma of the thyroid. A case confirmed by monoclonal antithyroglobulin antibody. AB - A 59-year-old woman with a history of papillary carcinoma of the thyroid gland developed three reddish nodules on the scalp. A skin biopsy showed a dermal tumor composed of sheets of clearly differentiated thyroid vesicles. Cutaneous metastases of thyroid carcinoma are very rare and this case is the first case confirmed by immunoperoxidase studies using monoclonal antithyroglobulin antibody. Positive reactions were obtained in colloid and at apices of thyrocytes. Monoclonal antibodies to human thyroglobulin may offer a unique opportunity to confirm the tissue origin of cutaneous metastasis. PMID- 2976002 TI - Residual plasma concentrations of acitretin (Ro 10-1670) and its metabolite (Ro 13-7652) after chronic administration. AB - In order to study the elimination of Ro 10-1670 (acitretin) and Ro 13-7652, its 13-cis-isomeric metabolite after chronic administration, determination of the residual plasma concentrations of these two compounds were assayed in 9 patients in a range of 22-30 days following cessation of a 2- to 7.5-month course with daily oral doses of 10-50 mg of acitretin. A highly sensitive HPLC method was used (2 ng/ml). Residual plasma concentrations of Ro 10-1670 were below the quantification limit in all cases. Residual plasma concentrations of Ro 13-7652 were below the quantification limit in 7 cases and respectively of 3 and 4 ng/ml in the other 2 cases. These data appear to confirm the absence of storage of acitretin even after an extended course of therapy. PMID- 2976003 TI - Cutaneous reactions to nifedipine. PMID- 2976004 TI - Serum DHAS in the maternal-fetoplacental system during the 28th-40th weeks of pregnancy. AB - The authors measured the serum levels of dehydroepiandrosterone sulfate (DHAS) in the maternal vein (MV), the umbilical vein (UV) and the umbilical artery (UA) during the 28th-36th weeks of pregnancy (n = 74) and in the 40th week (n = 34), to clarify the hormonal changes that occur between the maternal and fetal compartments. The following results were found: (1) The DHAS concentration increased significantly in MV, up to twice the concentration from the 28th-32nd weeks to the 33rd-36th weeks (p less than 0.01). From the 33rd-36th weeks to the 40th week it decreased significantly to one third of this value (p less than 0.01). DHAS levels revealed a decreasing tendency in the UA and UV serum from the 28th-32nd weeks to the 33rd-36th weeks, and there was a tendency to rise from the 33rd-36th weeks to the 40th week. (2) The serum DHAS values were found to be higher than in the MV, the UA-UV difference was not significant; however, the 'UA/UV X 100' value remained higher than 100% during the 28th-40th weeks. (3) The MV DHAS value showed slight correlation with the UV and UA DHAS concentration (r = 0.2951, p less than 0.01, n = 106; r = 0.2930, p less than 0.01, n = 100). There was a close correlation between the UV and UA serum DHAS levels (r = 0.8432, p less than 0.01, n = 98). The authors consider that the adrenal activity increases independently of the maternal adrenal cortex at term. PMID- 2976005 TI - Soluble mediators can replace helper T cells in the activation of resting B lymphocytes: evidence for a human B cell activating factor. AB - We were interested in studying the participation of T cell-derived soluble factors in the early steps of B cell activation. Thus supernatants containing such factors were obtained following activation of human T cell clones and their effects on isolated B cells investigated. These supernatants induced activation, blastogenesis and proliferation of purified resting human B cells. Our results strongly suggest the existence of a B cell Activating Factor (BCAF) of apparent molecular weight (m.w.) of 12,000-15,000 daltons which acts directly on resting B cells and replaces helper T cells in B cell activation. PMID- 2976006 TI - Studies on the murine interleukin-1 receptor. AB - The multiple biological actions of interleukin-1 (IL-1) on its diverse range of target tissues is consequent upon interaction of the cytokine with specific, high affinity cell surface receptors. In this report we describe the covalent crosslinking of natural porcine IL-1 and recombinant human IL-1 to intact cells of the murine EL-4 6.1 [NOB-1] thymoma subline, and the solubilization of functional receptors from these cells. Crosslinking studies revealed the existence of two polypeptides, of 100 Kda and 80 Kda, which are involved in IL-1 recognition. Chromatographic studies and ligand-blotting of the soluble receptor demonstrated that the smaller of these two polypeptides, which appears to be a glycoprotein, is capable of interaction with both forms of IL-1. PMID- 2976007 TI - Interdigestive gastroduodenal motor activity in subjects with increased gastric acid secretion. AB - The interdigestive gastroduodenal motor activity was studied in 7 patients with active duodenal ulcer and increased gastric acid secretion, in 7 patients with hypersecretory gastroduodenitis and in 7 subjects with normal acid secretion, in whom the increase in acid secretion was obtained by means of the intravenous administration of an H2 agonist, Impromidine. The gastroduodenal motor activity was recorded manometrically for about 200-300 min in basal conditions to obtain at least two subsequent activity fronts of the migrating motor complex (MMC cycle). Only in subjects with normal acid secretion was Impromidine administered at a dose of 2 micrograms/kg/h for 150 min followed by a dose of 10 micrograms/kg/h for another period of 150 min, to obtain, respectively, a submaximal and a maximal secretory response. Patients with spontaneous acid hypersecretion, with or without peptic ulcer, showed a longer than normal MMC cycle with a shorter than normal percent of time occupied by phase III. This motor activity was similar to that recorded in normal subjects during the increase in acid secretion induced by the lowest dose of Impromidine, whereas during the highest dose the gastroduodenal MMC was disrupted and replaced by an irregular motor activity. PMID- 2976008 TI - [Histochemical studies of rat lungs after the short-term influence of soil dust]. AB - There was examined the biological activity of soil dusts using histochemical methods. Intratracheal administration of dusts used in this study is a common method for testing activity of industrial dusts. The used soil dusts were characterized by high content of free silicon dioxide, 3 times higher than its content in dusts from power stations released in the process of burning coal and approximately to the amount in graphite dust. Both coal dusts and graphite dusts absorbed by the lungs cause silicoanthracosis. The investigations have been shown that soil dusts caused stimulation of the mitochondrial metabolism giving an increase of the activity of succinic dehydrogenase, lactic dehydrogenase, and ATPase induced by Mg++ ions. This was additionally confirmed by an increase of NADP activity which is an enzyme binding a chain or reactions regulating the hydrocarbonic metabolism. There was also observed an increased activity of the hydrolytic enzyme acid phosphatase. High activity occurred in the epithelium of bronchi and bronchioli and focally in pulmonary parenchyma. PMID- 2976009 TI - Molecular characterization of the mitochondrial DNA of a new stopper mutant ER-3 of Neurospora crassa. AB - An ethidium bromide-induced stopper mutant of Neurospora crassa is characterized at the molecular level. The mutant has two populations of mitochondrial DNA: a defective predominant mutant molecule and a basal level of the wild-type molecule. The aberrant DNA resulted after a 25-kbp deletion from the wild-type mitochondrial chromosome, which included major genes such as cytb, co1 and oli2. The deletion endpoints are located in the second intron of the ND5 gene, and in a sequence 250 nucleotides upstream of the co2 gene. The recombination has taken place between two nine nucleotide repeats CCCCGCCCC, one of which is close to a PstI palindrome at its 5' end. Thus the mutant ER-3 differs from all the other stopper mutants described previously in the extent and location of the deletions in the mtDNA. PMID- 2976011 TI - The role of the two E1a mRNA products of subgroup B adenoviruses in the regulation of early promoters of subgroup C adenoviruses. AB - HeLa cells were co-transfected with recombinant plasmids carrying adenovirus (Ad)2 or Ad3 E1a promoters fused to the chloramphenicol acetyl transferase gene (cat), and a plasmid encoding the Ad3 E1a promoter. Whereas no stimulating effect was observed on the Ad3 E1a promoter, the Ad2 promoter was inhibited. To determine which of the E1a gene products of Ad3 was responsible for the repressive effect, plasmids were constructed in which only the 13S or 12S mRNA product of Ad3 was expressed. Both the 12S and 13S mRNA products of Ad3 E1a were found to depress the transcription from the Ad2 E1a promoter. Each Ad3 E1a gene product was able to stimulate transcription from the Ad5 E2a early promoter in a manner similar to that of the Ad2 E1a gene products. In the case of the Ad5 E3 promoter, neither of the Ad3 E1a gene products stimulated transcription, but an inhibition was observed. These results suggest that both mRNA products of the Ad3 E1a region inhibit transcription at the TATA box transcription complex. PMID- 2976010 TI - Sequence analysis of the gtfC gene from Streptococcus mutans GS-5. AB - The nucleotide sequence of the gtfC gene, which codes for glucosyltransferase synthesizing both water-soluble and water-insoluble glucans, and its flanking regions from Streptococcus mutans GS-5, was determined. Although the gtfC gene (4218 bp) is preceded by a Shine-Dalgarno (SD) sequence, a promoter-like sequence for this gene could not be identified. The gtfC gene product composed of 1375 amino acid residues (approx. 153 kDa) is generally hydrophilic with three small hydrophobic domains. Two direct repeating units were found near the C terminus of the peptide. The gtfC gene has extensive homology with the previously sequenced gtfB gene. The homologous regions correspond to the signal sequence, an internal region, and the direct repeating units of the peptide. An open reading frame preceded by an SD sequence and followed by an inverted repeat sequence was found immediately downstream from the gtfC gene. The combined sequences of the gtfB and gtfC genes as well as flanking regions suggest that the two gtf genes and the small downstream coding region could be coordinately expressed within an operon. The possible evolution of the gtfC gene in S. mutans GS-5 is also discussed. PMID- 2976012 TI - Expression and secretion of human atrial natriuretic alpha-factor in Bacillus subtilis using the subtilisin signal peptide. AB - Using the signal peptide of the Bacillus subtilis subtilisin gene (aprE) and a synthetic cDNA corresponding to the mature region of the human atrial natriuretic alpha-factor (hANF), we have constructed a secretion vector. B. subtilis cells, when transformed with this vector, secrete immunoreactive hANF peptides into the medium at about 500 micrograms/liter. The hANF is the first human gene product to be secreted from B. subtilis using this signal peptide. We have used promoters active during vegetative growth or sporulation and hosts deficient in several extracellular proteases but some proteolysis of the secretion products still occurs. In addition, both cell growth and sporulation are adversely affected by hANF production. Possible explanations for this observation are inefficient secretion of the atrial hormone or toxicity of the precursor or mature peptide. PMID- 2976013 TI - Cloning of Thermomonospora fusca genes coding for beta 1-4 endoglucanases E1, E2 and E5. AB - Thermomonospora fusca chromosomal DNA was partially digested with EcoRI and fragments in the size range from 4 to 15 kb were isolated, ligated into lambda gtWES.lambda B arms, packaged, and the recombinant phages plated on Escherichia coli. The plaques were screened for carboxymethyl cellulase (CMCase) activity by a gel overlay procedure, and 25 plaques were positive among the 15,000 plaques that were screened. Positive phages were amplified and used to prepare infected E. coli extracts which were assayed for CMCase activity before and after treatment with antisera prepared against five purified T. fusca beta 1-4 endoglucanases (E1-E5). One phage produced an enzyme that was inhibited by E1 antiserum, nine of the phages produced enzymes that were inhibited by E2 antiserum, 14 produced enzymes that were inhibited by E5 antiserum and the enzyme produced by the other phages was not inhibited by any of the five antisera. The DNA insert present in the phage coding for E1 was cut into a number of different fragments which were subcloned into E. coli first using lambda gtWES.lambda B and then plasmid pBR322. The smallest active subclone, pTE12, contained a 3.1-kb insert. The insert present in one of the phages coding for E2 was also subcloned and the smallest active subclone pTE23 contained a 2-kb insert. E. coli HB101 containing plasmid pTE12 or pTE23 produced enzymes that were identical to E1 and E2, respectively, in all the properties tested. PMID- 2976014 TI - [Enterobacteriaceae infections among the population working in irrigated agricultural areas]. PMID- 2976015 TI - [Antibiotic-resistant strains of Klebsiella and Pseudomonas aeruginosa in coastal seawater]. PMID- 2976016 TI - [Use of the antibody neutralization reaction in evaluating the sanitary and hygienic condition of environmental objects in children's preschool institutions]. PMID- 2976017 TI - [Laparoscopy as a criterion of indications for the surgical restoration of fallopian tube patency]. PMID- 2976018 TI - [Surgical and conservative treatment of urinary incontinence in women. II. Surgical treatment]. PMID- 2976019 TI - [Local and systemic methotrexate for unruptured tubal pregnancy]. PMID- 2976020 TI - Compensation for occupational disease: hidden agendas. PMID- 2976022 TI - T-cell chronic lymphocytic leukaemia. PMID- 2976021 TI - Genetic linkage between lipoprotein(a) phenotype and a DNA polymorphism in the plasminogen gene. AB - Coronary heart disease risk correlates directly with plasma concentrations of lipoprotein(a) (Lp(a)), a low-density lipoprotein-like particle distinguished by the presence of the glycoprotein apolipoprotein(a) (apo(a)), which is bound to apolipoprotein B-100 (apoB-100) by disulfide bridges. Size isoforms of apo(a) are inherited as Mendelian codominant traits and are associated with variations in the plasma concentration of lipoprotein(a). Plasminogen and apo(a) show striking protein sequence homology, and their genes both map to chromosome 6q26-27. In a large family with early coronary heart disease and high plasma concentrations of Lp(a), we found tight linkage between apo(a) size isoforms and a DNA polymorphism in the plasminogen gene; plasma concentrations of Lp(a) also appeared to be related to genetic variation at the apo(a) locus. We found free recombination between the same phenotype and alleles of the apoB DNA polymorphism. This suggests that apo(a) size isoforms and plasma lipoprotein(a) concentrations are each determined by genetic variation at the apo(a) locus. PMID- 2976023 TI - Mg2+-dependent adenosine triphosphatase: an enzyme marker for ovine T lymphocytes. AB - Sheep T lymphocytes showed a cell surface magnesium-dependent adenosine triphosphatase (Mg2+-ATPase) reaction, which is reported to be characteristic of human B lymphocytes. In cryostat sections of lymph nodes, spleen and thymus, Mg2+ ATPase positive regions closely matched those labelled by sheep pan T monoclonal antibodies (Moab). An Mg2+-ATPase reaction was also found in fibroblastic recticulum cells of T cell regions in lymph nodes. Double labelling of cells from peripheral blood and peripheral lymph for Mg2+-ATPase and the pan T marker showed that 78% of the lymphocytes were positive for both of these markers. In cell suspensions enriched for B lymphocytes the percentage of cells positively labelled was decreased to 37%. Samples of each cell population which were labelled with a pan T Moab and analysed by flow microfluorometry revealed T cell levels which were not significantly different from those obtained by histochemical or immunohistochemical techniques. Less than 1% of lymphocytes positive for heavy and light chains of immunoglobulin (Ig) G were labelled with Mg2+-ATPase. Veiled cells in lymph and monocytes showed a cytoplasmic Mg2+-ATPase reaction. PMID- 2976024 TI - A single dose of UV radiation suppresses delayed type hypersensitivity responses to alloantigens and prolongs heart allograft survival in mice. AB - The systemic effect of ultraviolet (UV) irradiation on delayed type hypersensitivity (DTH), contact hypersensitivity (CHS) and allograft rejection was investigated in BALB/c mice which had been exposed to a single 1 h treatment with UV radiation (27 kJ/m2) from FS40 sunlamps (60% UVB). After UV irradiation (3-5 days), mice were treated on an unirradiated site with either a subcutaneous injection of allogeneic spleen cells or a topical application of the contact sensitizer oxazolone (OX). The DTH response to allogeneic cells and the CHS response to OX elicited 6 days after immunization were significantly lower in UV treated mice than in normal mice. Spleen cells from these animals were transferred intravenously into X-irradiated (600R) recipients which were immediately challenged with antigen and the DTH or CHS response elicited was determined 24 h later. Recipients of equal numbers of cells from sensitized and normal animals (6 X 10(6) from each donor) exhibited positive DTH or CHS responses to the antigen used to sensitize the donor. In contrast, recipients of equal numbers of cells from animals sensitized and UV suppressed to the same antigen showed a suppressed DTH or CHS response. This suppression was antigen specific. Treatment of cells from UV suppressed animals, prior to transfer, with complement and cytotoxic anti-Lyt 2 or anti-Thy 1.2 monoclonal antibodies abrogated the suppressive ability of these cells, in contrast to cytotoxic treatment with anti-L3T4 or anti-Lyt 1 monoclonal antibodies which had no significant effect. The suppressor cells therefore had the phenotype Thy 1.2+, Lyt 2+, L3T4-, Lyt 1-.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976025 TI - [Role of Malassezia furfur in seborrheic dermatitis in adults and therapeutic efficacy of fenticonazole]. PMID- 2976026 TI - [Longitudinal study of a selected population with occupational allergic contact dermatitis]. PMID- 2976027 TI - Comparison of fenoterol, ipratropium bromide and their combination in patients with bronchial asthma. PMID- 2976028 TI - Mesodermal band deformity and associated pathologies. Case report. PMID- 2976029 TI - Genes for Down syndrome and Alzheimer disease share the same chromosome 21. PMID- 2976030 TI - Late and delayed problems of Down syndrome patients. PMID- 2976031 TI - Myocardial dysfunction in birth asphyxia. PMID- 2976032 TI - Coexpression of glial fibrillary acidic protein and vimentin in the central and peripheral nervous systems of the twitcher mutant. AB - A method to purify glial fibrillary acidic protein (GFAP) from mouse spinal cord is described, which permits the measurement of GFAP in the sciatic nerve of the twitcher mutant and control mouse. Cytoskeletal proteins from sciatic nerves and purified GFAP standards were electrophoresed on gel, transferred to nitrocellulose paper, and immunostained with anti-GFAP antibody. From the immunostained, 51,000-dalton band, we estimated about 200 ng GFAP per 50 micrograms of cytoskeletal protein in the twitcher sciatic nerve. The control nerve showed no detectable GFAP. Double-labeled fluorescence immunocytochemistry showed that in the brainstem of twitcher mutant, GFAP and vimentin were coexpressed in the majority of astrocytes. PMID- 2976033 TI - Growth factors for human glial cells in culture. AB - Using a new double-labeling immunofluorescence technique, we assessed various growth factors on their ability to promote proliferation of cultured human glial cells. Cells studied were fetal astrocytes, fetal Schwann cells, adult astrocytes, and adult oligodendrocytes. Effective agents for fetal astrocytes were glial growth factor from the bovine pituitary, platelet-derived growth factor, fibroblast growth factor, and 4 beta-phorbol 12,13-dibutyrate. For fetal Schwann cells, mitogens were glial growth factor from the bovine pituitary, platelet-derived growth factor, nerve growth factor, and 4 beta-phorbol 12,13 dibutyrate. Adult astrocytes and oligodendrocytes did not normally divide in culture, and none of the agents tested were effective in inducing their proliferation. The report that interleukin-2 was a mitogen for oligodendrocytes could not be replicated in the present study on any of the glial cell types. PMID- 2976034 TI - Structural complexity of primary cultures of astrocytes as revealed by transverse sections. AB - The tissue architecture of low-, medium-, and high-density primary mouse astroglial cultures was examined in horizontal and transverse planes using the electron microscope. It was found that the low-density (colony) cultures consisted of a true monolayer, whereas the medium- and high-density (confluent) cultures consisted of anywhere from two to seven overlapping sheets enclosing a substantial intercellular space. The presence of these multiple overlapping sheets in confluent astrocyte cultures should therefore be taken into consideration when interpreting data of cell-membrane-related phenomena such as ion fluxes. PMID- 2976035 TI - Tetanus toxin binding to isolated and cultured rat retinal glial cells. AB - The presence of immunocytochemically detectable membrane receptors for tetanus toxin, supposedly composed of higher gangliosides, is widely accepted as a marker of neuronal cells. We now demonstrate that Muller cells, a unique glial cell type of the vertebrate retina, possess specific tetanus toxin (TT)-binding sites. Single cell suspensions were prepared from adult rat retina by a gentle dissociation method, and the Muller cells, unequivocally identified by their morphology, could be immunocytochemically double-labeled by antisera to vimentin and to TT. The expression of complex gangliosides by identified Muller cells was also demonstrated by immunofluorescence labeling with the monoclonal antibody A2B5. Using the double-immunolabeling method for the identification of Muller cells we show that specific tetanus toxin binding is acquired by these cells during postnatal maturation both in vivo and in vitro. In vivo the percentage of tetanus toxin-positive Muller cells increases from 0% in 4-day-old animals to 10% on postnatal day 8, reaching the adult level of about 95-100% around day 30. In retinal monolayer cultures prepared from newborn rats, the majority (65%) of vimentin-positive non-neuronal cells became TT-positive during a 2-week culture period, indicating that this population of non-neuronal cells represents differentiating Muller cells. Again, comparable results were obtained with A2B5, supporting the conclusion that Mullerian glia expresses surface molecules, which are normally regarded as neuronal markers. PMID- 2976036 TI - Direct effects of carbachol on membrane potential and ion activities in leech glial cells. AB - Ion-selective double-barreled microelectrodes were used to measure the activities of intracellular K+ and Na+ (aKi, aNai) and the membrane potential (Em) in neuropile glial cells as well as extracellular K+ (aKe) in the neuropile of segmental ganglia in the leech, Hirudo medicinalis. Bath-application of carbachol resulted in a prominent membrane depolarization. This depolarization was accompanied by transient increases of aNai and aKe, whereas aKi decreased. It is suggested that the carbachol-induced depolarization and the underlying ion activity changes are due to activation of an acetylcholine receptor-coupled cation channel in the membrane of the neuropile glial cells. PMID- 2976037 TI - The perinodal astrocyte. AB - Several studies have demonstrated the presence of perinodal astrocyte processes at nodes of Ranvier in the central nervous system, suggesting that, in addition to the axon and oligodendrocyte, astrocytes participate in the formation of mature central nodes. The specific association between perinodal astrocyte processes and nodal membrane develops at the time of, or soon after, the appearance of relatively differentiated nodes of Ranvier. This interaction is likely to be mediated by cell adhesion molecules. J1 is a member of a family of glycoproteins that share a common carbohydrate epitope, designated L2/HNK-1, and that have been implicated in cell-cell interactions. This glycoprotein is concentrated at the interface between perinodal astrocyte processes and the nodal region of the axon. Moreover, N-CAM, which is a member of the same family as J1, and cytotactin, an extracellular matrix component produced by glia, are localized at the interface between the axon and perinodal astrocyte processes at nodes of Ranvier. The association of perinodal astrocyte processes with nodal membrane in the central nervous system is similar to that exhibited by perinodal Schwann cell processes at peripheral nodes, and similar functional properties have been suggested for these two glial cell processes, including production of nodal gap substance, buffering of perinodal extracellular ion concentration, and development and/or maintenance of nodal specializations in the axon membrane. Perinodal astrocyte and Schwann cell processes may also function as extraneuronal sites for the synthesis of voltage-sensitive sodium channels, to complement neuronal perikaryal synthesis and axonal transport. Ultrastructural studies on specialized patches of axon membrane within some unmyelinated, demyelinated, and dysmyelinated axons support the hypothesis of a specific role for perinodal astrocyte processes in the assembly, stabilization, and/or maintenance of axolemma with nodal characteristics. These observations suggest a multiplicity of functions for perinodal astrocyte processes at central nodes and implicate the astrocyte as an important component of the node of Ranvier. PMID- 2976038 TI - Cytotopographical specialization of enzymatically isolated rabbit retinal Muller (glial) cells: K+ conductivity of the cell membrane. AB - Muller (radial glial) cells were isolated from rabbit retinae by means of papaine and mechanical dissociation. Regional membrane properties of these cells were studied by intracellular microelectrode recordings of potential responses to local application of high K+ solutions. When different parts of the cell membrane were exposed to high K+, the amplitude of the depolarizing responses varied greatly, indicating a strong regional specialization of the membrane properties. Using morphometrical data of isolated rabbit Muller cells, and a simple circuit model, we calculated the endfoot membrane to constitute more than 80% of the total K+ conductance of the cell; the specific resistivity of the endfoot membrane was about 400 omega cm2, i.e., more than 40 times less than that of the membrane of the vitread process, which is immediately adjacent. This kind of regional membrane specialization seems to be optimized in respect to the Muller cells' ability to carry spatial buffering K+ currents. PMID- 2976039 TI - Model of potassium dynamics in the central nervous system. AB - A one-dimensional numerical model of potassium dynamics in the central nervous system is developed. The model incorporates the following physiological processes in computing spatial and temporal changes in extracellular K+ concentration, [K+]o: 1) the release of K+ from K+ sources into extracellular space, 2) diffusion of K+ through extracellular space, 3) active uptake of K+ into cells and blood vessels, 4) passive uptake of K+ into a cellular distribution space, and 5) the transfer of K+ by K+ spatial buffer current flow in glial cells. The following tissue parameters can be specified along the single spatial dimension of the model: 1) the volume fraction and tortuosity of extracellular and glial cell spaces, 2) the volume fraction of the cellular distribution space, 3) rate constants of active uptake and passive uptake processes, and 4) glial cell membrane conductance. The model computes variations in [K+]o and current flow through glial cells for three tissue geometries: 1) planar geometry (the retina and the surface of the brain), 2) cylindrical geometry (tissue surrounding a blood vessel), and 3) spherical geometry (tissue surrounding a point source of K+). For simple sources of K+, the performance of the model matches that predicted from analytical equations. Simulations of previous ion dynamics experiments indicate that the model can accurately predict ion diffusion and K+ current flow in the brain. Simulations of electroretinogram generation and K+ siphoning onto blood vessels suggest that unanticipated K+ dynamics mechanisms may be operating in the central nervous system. PMID- 2976040 TI - Ultrastructural study of phagocytic activities of young astrocytes in injured neonatal rat brain following intracerebral injection of colloidal carbon. AB - The cellular reaction to injury in the mature central nervous system (CNS) has been extensively studied in both man and animals, while a detailed study of the reaction of the immature CNS to injury is lacking in the literature. This study was undertaken to elucidate the response of young astrocytes following injection injury to developing brain. Colloidal carbon was applied because it is a suitable marker for phagocytosis, it is nontoxic, and it is readily identifiable by light and electron microscopy. The cerebral cortex of the neonatal rat was injected with 0.1 microliter of colloidal carbon solution. The animals were allowed to survive from 1 hour to 30 days postoperation. The brains were fixed by vascular perfusion and processed for light and electron microscopy. Carbon particles were ingested in membrane-bound vacuoles and sequestered in lysosomes of young astrocytes. Astrocytes, loaded with carbon particles, were identified after 4 days, and were seen in abundance between 10 to 21 days postoperation. Carbon laden astrocytes were seen in the immediate vicinity of the site of the injection; in the surrounding, apparently normal, neuropil; and in the perivascular regions. This study demonstrates the ability of young astrocytes to engulf foreign particles injected into the developing brain. The presence of carbon particles in astrocytes located further away from the site of injection is discussed. PMID- 2976041 TI - Swelling of astrocytes causes membrane potential depolarization. AB - Rat brain astrocytes growing in primary monolayer cultures were swollen by exposing them to media of decreasing osmolality caused by removal of NaCl, and the effects of this treatment on their membrane potentials were measured by intracellular recording. Depolarizations were seen that were proportional to the degree of swelling, reaching a maximum of around 60 mV when 80-100 mM NaCl was removed from the reaction media, which had an original total osmolality of 290 mosmolal. These effects were completely reversible, since restoring the cells to iso-osmotic medium after a 2-min exposure caused an immediate repolarization back to the original membrane potentials, and depolarizations were not seen when isotonicity was maintained by replacing NaCl with sucrose. Partial repolarization was seen during an extended period (30 min) of exposure to hypo-osmotic medium, mirroring a regulatory volume decrease we have previously described in these cells under identical conditions. In ion-replacement studies depolarizations were seen when the solution was made hypo-osmotic with the large cation N-methyl-D glucamine totally replacing Na+ in the medium. Removal of Cl- from the medium also had no effect on the initial swelling-induced depolarization. These results show that even moderate swelling of astrocytes in primary culture results in marked depolarization of their membrane potentials; possible mechanisms for this effect and the potentially profound implications for the swelling of astrocytes seen in situ under pathological conditions are discussed. PMID- 2976042 TI - Schwann cell myelination of the myelin deficient rat spinal cord following X irradiation. AB - The myelin-deficient (md) rat is an X-linked myelin mutant that has an abnormality of oligodendrocytes and a severe paucity of myelin throughout the CNS. This lack of myelin makes it an ideal model in which to study the cellular interactions that occur when "foreign" myelinating cells are induced in the milieu of this nonmyelinated CNS. In this study, Schwann cells were induced in the lumbosacral spinal cord by exposing it to radiation, a technique demonstrated repeatedly in other nonmutant strains of rats. Md rats and their age-matched littermates were irradiated (3,000 to 4,000 R) at 3 days of age and perfused 16 22 days later after pulse labeling with tritiated thymidine. In the md rat, Schwann cell invasion progressed from the area of the spinal cord-nerve root junction and extended into the dorsal columns and adjacent gray matter. Autoradiographic evidence revealed that many of these cells incorporated 3H thymidine, indicating that they were undergoing proliferation. Ultrastructural observations showed that there was an integration of these intraspinal Schwann cells with the cells normally occurring in this environment, i.e., oligodendrocytes and astrocytes. The extent of migration and division of Schwann cells, as well as their interactions with glial cells, were similar to those seen in the nonmutant irradiated littermates. These studies provide conclusive evidence that md rat axons are normal with respect to their ability to provide trophic and mitogenic signals to myelinating cells. PMID- 2976043 TI - Malformation of the head in Down syndrome. PMID- 2976044 TI - Competition and regulation redux. PMID- 2976045 TI - The emphasis on measurement in quality assurance: reasons and implications. AB - Quality assurance initiatives and studies have focused more on the measurement of quality than on quality improvement and control. The bias toward measurement of current efforts is attributable to the marked preference for a competitive approach to quality assurance, in which measures play a central, triggering role. Reinforcing this bias is a long-standing belief that too little is known about measuring or even defining quality. This paper examines and evaluates these rationales. It concludes that a strong case can be made for pursuing a different overall strategy, one more balanced toward quality improvement and control and, within quality measurement itself, one less focused on the kinds of global outcome measures currently associated with the competitive approach to quality assurance. PMID- 2976046 TI - Standards of care in medicine. AB - Growing concern over quality of care, increasing evidence of widespread differences in medical practice, increasing realization that much of medical practice lacks rigorous scientific underpinning, and a growing perception that society is not receiving sufficient value for resources expended on health care have led many to make the case for establishing rigorously constructed standards of care. After assessing deficiencies in the current process for generating standards of care and for implementing them, this article elaborates a two-step process for collecting clinical research data and incorporating expert clinical judgment. I believe a concerted effort to systematically develop medical standards of care should be made and that properly constructed and properly maintained standards will be acceptable to physicians. PMID- 2976047 TI - An ethical model for improving the patient-physician relationship. AB - Working with the ethical principles of beneficence and autonomy, I address three questions: How can communication between physicians and their patients be improved? How does the public availability of practice guidelines and other alternative sources of health information influence the interactions between physicians and patients? How can a partnership in health care in which the patient and the physician share responsibility be achieved? Educating patients for a partnership in health care will offer a further incentive for physicians to enter and sustain such a partnership. PMID- 2976048 TI - National forecasts of the medical care costs of AIDS: 1988-1992. AB - Two statistical extrapolation models and data on the number of AIDS cases reported between January 1984 and June 1988 yield projections on the future numbers of AIDS cases which are considerably higher than those of earlier projections, while analysis of the lifetime cost of treating an AIDS patient in 1988 dollars yields estimates lower than most previous estimates. Based on an estimate of $60,000 per patient holding between 1988 and 1992, the analysis forecasts the cumulative lifetime medical care costs of treating all AIDS patients diagnosed with AIDS to be about $2.6 billion in 1988, $3.5 billion in 1989, $4.7 billion in 1990, $6.0 billion in 1991, and $7.5 billion in 1992. PMID- 2976049 TI - Medicare spending and mortality rates of the elderly. AB - This paper addresses the issue of Medicare expenditures and mortality rates of the elderly by examining cross-sectional area data for 1980. Econometric methods are used to determine whether age-, sex-, race-specific mortality rates for elderly population cohorts are lower in areas where Medicare expenditures per beneficiary are higher, controlling for the effects of differences in population and area characteristics. The results show that greater Medicare spending per beneficiary is associated with significantly lower mortality rates. Furthermore, the level of Medicare spending is positively related to the level of public income support for the elderly and the extent of the elderly's contact with hospitals and nursing homes. PMID- 2976050 TI - Using patient age in defining DRGs for Medicare payment. AB - Major changes were recently implemented in the diagnosis related groups (DRGs) used for payment under Medicare's prospective payment system (PPS). As of October 1987, patient age was no longer used in combination with the presence of a complication or comorbidity (CC) to define DRGs. We present the results of one of the studies that led to this change. We analyzed inpatient charges for 1984 and found that the use of age in combination with CC is inappropriate for grouping Medicare patients. The original DRGs resulted in an underpayment for CC patients and an overpayment for patients 70 years or older without a CC. PMID- 2976051 TI - The dynamics of Medicaid enrollment. AB - This longitudinal study examines transitions on and off Medicaid in the 1984 Panel of the Survey of Income and Program Participation. A majority of those enrolled at the outset, but just 43% of those enrolled at any time during the 32 month survey, remained on Medicaid throughout. While slightly less than half of those departing the program subsequently enjoyed improved employment, private insurance, and higher incomes, nearly half were still poor and 55% became uninsured, indicating that persons who lost their Medicaid cards were in real danger of being without insurance and financial access to health care--a serious disincentive to get off welfare. PMID- 2976052 TI - The impact of alternative hospital payment systems on Medicaid costs. AB - From 1982 onward, 25 states adopted three broad strategies to control their Medicaid hospital costs: multiple-payer rate setting, Medicaid-only prospective payment, and selective contracting. In this analysis of the three approaches, I found that the multiple-payer systems were particularly effective at controlling Medicaid payments because of their ability to control statewide hospital costs. The Medicaid-only prospective payment systems were also successful, but appear to be effective only in the short term. California's Medicaid-only selective contracting program was also successful, largely because of the simultaneous implementation of cost-control incentives for all hospitals in the state. I also found sharp reductions in Medicaid admissions where rates were set low relative to costs, as well as a shift in Medicaid admissions to public hospitals in Medicaid-only prospective payment states. PMID- 2976053 TI - An evaluation of the information interface in the design of computer-driven aids for expressive communication. AB - Computer-based systems as assistive aids for expressive communication have become increasingly sophisticated. One such system, which provides great flexibility in the specification of an operational configuration and which is highly user oriented in its design, is discussed here. In particular, attention is directed towards the relationship between the physical signals which constitute the representation of graphic-symbol communication units and the transfer of conceptual information which these symbols ultimately mediate. This relationship is investigated practically with a view to establishing a connection between operational criteria at the functional level and the supporting physical infrastructure at the computational level. This investigation leads to some conclusions not only in terms of design strategy for flexible communication aids, but points to issues of broader significance in the area of computer-based assistive devices for those with special needs. PMID- 2976054 TI - Revascularization by percutaneous transluminal coronary angioplasty: how much is enough? PMID- 2976055 TI - Parliament and disability in Britain. AB - There is a large number of people with disabilities in Britain, yet politically disability issues have a low profile. This paper presents evidence from a study of the attitudes to welfare of 96 Members of Parliament (MPs) illustrating that their general level of awareness of future challenges to policy is low. Overall, whether at a general level or when questioned about specific policy preferences, the needs of people with disabilities rarely figured in MPs' responses. Yet, despite this, the passage of the private members' Disabled Persons' (Services, Consultative and Representative) Act, 1986 shows that there is some prospect both for legislative change and for raising further MPs' awareness of the needs of people with disabilities. PMID- 2976056 TI - Cost and quality of psycho-pedagogic institutional care: the case of 56 establishments in the Swiss canton of Vaud. AB - We studied 56 boarding institutions caring for 834 children and adolescents with different kinds of difficulties or handicaps in the Swiss Canton of Vaud. In order to weigh the purely qualitative approach we performed two different statistical analyses: 1) a correlation matrix between all qualitative and quantitative variables with the daily individual cost of the institutional care. This cost has been calculated in a standard manner and independently of each institution's accounts. 2) a factor analysis of the collected data led us to the identification of different patterns of institutional functioning. This study stresses the importance of the staff/pupil coefficient, i.e. the richness of the pedago-therapeutic framing--the intervention of specialists such as special teachers, psychologists, and speech and/or motor therapists-, and more particularly the higher ratio of specialist help compared to pure educative staff. The severity of the cases, evaluated by independent specialist observers, was in direct relation to this factor independently of the kind of difficulty or handicap. PMID- 2976057 TI - Disability in full-length feature films: frequency and quality of films over an 11 year span. AB - The purpose of this study was to assess frequency of disabling conditions in feature films and the general quality of these films (as judged by professional film critics reviews and movie going consumer ratings) over an 11-year span. 'Psychiatric disorder' were most frequently depicted. Professional film critics were markedly more negative in their reviews than ratings made by movie going consumers. PMID- 2976058 TI - [Handicaps and maladjustment: evaluation of social policies]. AB - This article examines various methods of assessing social policies with regard to the integration of handicapped or maladjusted children and adults in France. It begins with an account of the existing system of policies and services for handicapped or maladjusted children and adults in France today. By following the progress of an educationally retarded child (with minor mental handicap or retardation plus behavioural problems), noting the various instructions with which the child is involved, the reader will come to realise the limited nature of the facts available concerning the effectiveness of social integration schemes for the handicapped and maladjusted. The paper then examines the statistical data issued by the ministries of health and education as well as from separate institutions. These data indicate a lack of awareness of the various passages from childhood to adulthood, a bridge between institutions coinciding with the passage into adult life. Educational statistics are completely independent of other statistics on adult services. Measurements focus on institutional activity and often ignore the effects on the individual of the various measures taken. In 1975, France passed legislation concerning integration but little has been done to evaluate the effects of this law. Based on observations in one department of France, the author has found unequal development of the policy for educational integration on the one hand and vocational rehabilitation on the other. Such inequalities are producing increasing difficulties. Integration of physically handicapped children into an ordinary school environment has met with some success. However educational maladjustment, as manifested by children with minor mental handicap or retardation and those with behavioral problems, remains largely beyond the scope of legislation on educational integration. Increasingly, educationally maladjusted children are sent so special schools. Since job opportunities for the handicapped and maladjusted are in short supply, employment inequalities exist. As a result transitional measures or other programmes designed to occupy time have increased whereas employment in ordinary work settings has declined. More often than not, women are employed in "Centres d'Aide par le Travail" (Employment centres for the handicapped). Access to ordinary jobs tends to be mainly available to men whereas specialised employment is mainly for women.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2976059 TI - Cervical cancer risk and use of depot-medroxyprogesterone acetate in Costa Rica. AB - The relationship between cervical cancer and the use of depot-medroxyprogesterone acetate (DMPA) was examined in a nationwide case-control study in Costa Rica. Cases were women ages 25-58 years of age with invasive squamous cell cancer (N = 149) or carcinoma in situ (CIS, N = 415) reported by the National Tumor Registry during 1982-84. Controls (N = 764) were randomly selected during a nationwide household survey. Using logistic regression, we adjusted for known risk factors for cervical cancer. DMPA use was associated with a risk of CIS of 1.1 (95% confidence interval 0.6-1.8) and a risk of invasive cancer of 1.4 (95% confidence interval 0.6-3.1). The slightly elevated risks observed may be the result of chance or a detection bias. One limitation of this study is that few women had used DMPA for longer than two years. PMID- 2976060 TI - Transmission of hepatitis B virus infection in Ethiopia with emphasis on the importance of vertical transmission. AB - This two-year controlled study was conducted to define the mode of transmission of hepatitis B infection in Ethiopia. Twenty-five of 500 (5%) expectant mothers were identified as HbsAg positive (Group A). A control group of 29 mothers negative for HBsAg (Group B) was randomly selected. These 54 mothers and their families (husbands, infants and the infants' older siblings) were followed for two years with determinations of serological markers of ongoing or past hepatitis B virus infection at 6, 12 and 24 months. Social and clinical data as well as possible factors contributing to the spread of HBV (ie tattoos, circumcision, etc.) were compiled from questionnaires. Twenty-one of the 25 (84%) HBsAg positive mothers had anti-HBe and none had HBeAg. One case of vertical hepatitis B virus transmission was seen while horizontal transmission during the study period occurred in two infants and in two older siblings. Potential risk factors for non-perinatal transmission were highly prevalent in both groups and may represent the main route of transmitting the infection, while, as shown in the present study, the vertical transmission of HBV infection only plays a minor role in the maintenance of the high infection rate in Ethiopia. Vaccinating all babies soon after birth would, therefore, be an effective means of eradicating or reducing the hepatitis B virus infection and its sequelae. PMID- 2976061 TI - Changes in lymphocyte subpopulations in peripheral blood of splenectomized patients. AB - Twenty-five patients who had undergone splenectomy were investigated. Splenectomy was performed in 16 patients because of traumatic rupture of the spleen, in 6 due to idiopathic thrombocytopenic purpura (ITP) and in 3 because of other causes. The absolute numbers of T-cell subclasses--helper/inducer (CD4+) and suppressor/cytotoxic (CD8+)--and of B-cells were determined in peripheral blood. The CD4+/CD8+ was decreased in 7 patients and inverted in 10. The change in the ratio was not due to a decreased number of CD4+ cells but to an increase of CD8+ cells. Sex, age, and the time at which splenectomy was performed bore no relation to the CD4+/CD8+ ratio. In 13 of the 25 patients a significant increase in the number of polyclonal B-cells was found. None of the patients had severe infections. These observations suggest that splenectomy is responsible for the change in the CD4+/CD8+ ratio. Susceptibility to infections seems to correlate with the absolute decrease of CD4+ cells rather than with the change of the ratio. PMID- 2976062 TI - Experimental sclera-implanted keratoprosthesis. AB - An experimental design of keratoprosthesis anchored into the limbal sclera is presented. The surgical procedure enables a watertight junction between the keratoprosthesis and the globe. Dehiscence of the keratoprosthesis-limbal sclera junction occurs when an area of locus resistentia minoris is created in the lower limbus. An artificial drainage system is necessary for the durability of such a keratoprosthesis. PMID- 2976063 TI - Therapeutic prospects of THF--a thymic hormone--in autoimmune processes. PMID- 2976064 TI - A sensory evoked potential comparison of persons 'at risk' for Huntington's disease and hospitalized neurotic patients. AB - Scalp-recorded sensory evoked potentials (EPs) elicited by left and right median nerve stimulation (LSEP and RSEP), checkerboard pattern flash (VEP) and acoustic click (AEP) were obtained in 22 individuals 'at risk' (AR) for Huntington's disease and 22 hospitalized neurotic patients matched for age, gender and intelligence. EPs of AR subjects were generally similar to those of the neurotic comparison group, in terms of overall configuration; however, mean amplitudes were significantly lower for ARs. While the general amplitude reduction for ARs was seen in all three modalities, the somatosensory modality yielded the most abnormal findings. PMID- 2976065 TI - Keratan sulphate and the ultrastructure of cornea and cartilage: a 'stand-in' for chondroitin sulphate in conditions of oxygen lack? AB - Corneas from mouse, rat and rabbit were analysed quantitatively and/or qualitatively for collagen and acid glycosaminoglycans. They were examined by light and electron microscopy, using Alcian blue and Cupromeronic blue, in critical electrolyte concentration methods, with or without digestion by hyaluronidase, chondroitinases and keratanase, for their sulphated glycosaminoglycan distributions. Glycosaminoglycan patterns were very different in the three species. Mouse lacked chemically detectable keratan sulphate, which was present in considerable amounts in rat and rabbit stroma. Mouse corneal stroma proteoglycan filaments were located predominantly at the gap zone of the collagen fibrils, mainly at the d band, with few at the a and c bands. Rat and rabbit micrographs were more complicated, with many proteoglycan filaments at the a and c, as well as the d and e bands. These findings support the proposal that the a and c bands were specific binding sites for keratan sulphate proteoglycan (Scott & Haigh, 1985b). Evidence from studies on cornea and cartilage suggests that keratan sulphate, rather than chondroitin sulphate is produced in conditions of O2 lack. Metabolic mechanisms which could account for this balance are proposed The production of uridine diphosphate glucuronic acid is the key step, which is sensitive to hypoxia, lactate and NAD:NADH ratios. PMID- 2976067 TI - Liquid chromatographic analysis of doxazosin in human serum with manual and robotic sample preparation. AB - A specific method for the determination of the antihypertensive drug doxazosin in human serum is described. The method utilizes the related drug prazosin as an internal standard and is based on a simple extraction scheme followed by analysis by reversed-phase ion suppression high-performance liquid chromatography (HPLC) on an alumina-based column with fluorescence detection. The method is completely automated with a flexible robotic system for the analysis of drugs in biological fluids. The robotic automation of the method allows a 20% increase in the sample throughput and the savings of about 7 man-hours a day. Both the manual and robotic procedures yield precise quantitative results over the therapeutically relevant concentration range of 0.5 to 20 ng/mL of serum. PMID- 2976066 TI - Training students with profound or multiple handicaps to make requests via microswitches. AB - In a series of three experiments, we evaluated the use of microswitches as a means for students with profound, multiple handicaps to demonstrate preferences between toys and to make requests for specific activities. In Experiment 1, 5 students learned to demonstrate toy preferences by using microswitches to activate battery-operated toys. Experiment 2 was conducted to evaluate the students' preferences for social attention. Microswitches were used to activate prerecorded messages that signaled the classroom teacher to attend to the students. In Experiment 3, the students used the switches and prerecorded messages to make specific requests of educational staff in school and community settings. Results of these experiments, evaluated within multiple baseline, alternating treatments, and simultaneous treatments designs, indicated that these students could request specific activities. Results are discussed with respect to the continued use of microswitches and to program development. PMID- 2976068 TI - Educational readiness of survivors of neonatal encephalopathy associated with birth asphyxia at term. AB - Standardized individual psychoeducational school-readiness tests were completed by 127 nondisabled survivors of moderate (lethargy, hypotonia, and suppressed primitive reflexes) or mild (hyperalertness, hyperexcitibility) neonatal encephalopathy associated with term birth asphyxia. Application of the readiness test battery to the children with moderate or mild encephalopathy and a peer population, revealed that children with moderate encephalopathy had significantly lower scores for many tests than those in the other groups. Children with mild encephalopathy performed well. We found no significant differences due to social variables. For the moderate group, we found an increased number of types of anticonvulsants and abnormal findings on the neurologic examination at neonatal intensive care unit (NICU) discharge to be predictors of low achievement on school-readiness tests. Also for the nondisabled moderate group, multiple regression analysis of the independent variables with the 5.5-year scores added variables suggestive of intrauterine growth retardation to the prediction of lower scores on many school-readiness tests. We conclude that clinical categorizing of moderate neonatal encephalopathy associated with birth asphyxia in term infants selects a group of children with an increased percentage of school-readiness delay, and could be a useful indicator for clinicians and educators in defining those neonates who may need special preschool evaluation and benefit from a modified early school program. PMID- 2976069 TI - An investigation of the dependence of severely disabled people in a hospital. AB - This paper describes a pilot study of the dependence of 20 severely disabled patients on nursing staff in a long-term rehabilitation ward. Dependence was measured as the number of 15-minute periods spent by a patient in receipt of various categories of nursing care during fixed observation times. Measurements were made by patient activity sampling and were in reasonably good agreement with the results of nurse activity sampling. Severity of disability gradings of the International Classification of Impairments. Disabilities and Handicaps (ICIDH) were adapted to give an overall disability score for each patient. Appropriately summarized numerical data are presented in the form of tables and bar charts. Patient feeding was firmly established as the most time-consuming of the direct nursing care activities. There was also a high statistical correlation between feeding dependence and severity of disability and this finding is discussed at some length. PMID- 2976070 TI - Desmopressin acetate prophylaxis in a patient with hemophilia A: report of case. AB - Derivatives of an antidiuretic hormone raise factor VIII levels in patients with mild or moderate hemophilia A. This case report describes the use of desmo pressin acetate to raise the factor VIII level in a patient with hemophilia A, before and after extraction of 27 teeth. PMID- 2976071 TI - Use of dental X rays on postirradiated patients with head and neck cancer. AB - As cancer therapy becomes more successful and cancer survival rates increase, the dentist will be treating more patients who have received radiation therapy for head and neck cancer. Occasionally, patients and health professionals have indicated a belief that patients who have received irradiation to the head and neck regions should not be subjected to additional radiation through dental diagnostic X-ray exposures. A literature search failed to find any references that specifically addressed this question. This study reflects the opinions of 278 radiation oncologists (400 surveyed) who responded to questions about contraindications of dental X rays for the patient with head and neck cancer. PMID- 2976072 TI - Adrenergic nerve function and contractile activity of the caudal artery of the streptozotocin diabetic rat. AB - The adrenergic nerve function and contractile responses of the densely innervated caudal artery of the 8-week streptozotocin (SZ) (65 mg/kg i.p.) diabetic rat were investigated. Segments of this artery were removed from diabetic and control rats, placed in Krebs-filled tissue baths (37 degrees C) and isometric tension recorded. Contractile responses to sympathetic nerve activation by electrical stimulation and to cumulative concentrations of norepinephrine (NE) and tyramine were recorded. In order to determine NE content, the NE was extracted from the caudal artery, isolated by adsorption chromatography, and quantified by HPLC with electrochemical detection. NE accumulation and release were also studied by quantifying the amount of tritiated NE [( 3H]NE) and its metabolites in extracts of the tissue or incubation medium. The responses of the caudal artery of SZ diabetic rats to electrical stimulation (4-16 Hz) and to tyramine (1 X 10(-5)-1 X 10(-4) M) were significantly less than those of arteries from control rats and the NE content reduced by 41%, while sensitivity to NE was unchanged. Diabetic arteries also accumulated and released more [3H]NE than did arteries from control rats. These results establish that neurovascular function of the isolated caudal artery of the 8-week SZ diabetic rat is abnormal and suggest that problems in the ability of adrenergic nerves to store and release NE may contribute to this dysfunction. Such changes may play a role in the cardiovascular disturbances associated with diabetic autonomic neuropathy. PMID- 2976073 TI - Echocardiographic evaluation in acromegalic patients. PMID- 2976074 TI - Active tendon implants in flexor tendon reconstruction. AB - Forty-five active flexor tendon implants were evaluated after placement in scarred tendon beds of digits II through V. The implant is constructed of silicone rubber with a Dacron core, terminating in a loop proximally and a metal plate distally. Modification of the implant during the period of study has improved its reliability and longevity. The improvement in total active motion (TAM) averaged 72 degrees during implant functioning (stage I) in a group of digits that before operation were classified as 78% Boyes grade 5 (salvage). Complication rate during stage I was 11% (5 out of 45). Of the 27 digits evaluated after implant replacement by tendon autograft (stage II), there was an overall improvement in 62 degrees total active motion with 70% of digits being Boyes grade 5. Many of the complications were believed to be avoidable with experience. This study demonstrates the feasibility of an active tendon implant and the possibility of a permanent prosthesis. PMID- 2976075 TI - Management of health care workers remotely vaccinated for hepatitis B who sustain significant blood and body fluid exposures. PMID- 2976076 TI - Characterization of the avian T cell receptor. PMID- 2976078 TI - [Measurement of Gla proteins by dot method and clinical study (protein S, protein C, and prothrombin)]. AB - In this study, as a new assay method for Gla protein antigens, the "dot immunobinding assay" which is used on screening for a large number of antibodies (Dot method) was applied. Plasma protein S(PS), protein C(PC) and prothrombin were measured in normal and toxemic pregnant women by the "dot method". The results obtained are as follows. 1. The PS concentration in the third trimester was significantly decreased, with a count of 90.0 +/- 18.6%. 2. PC in the first trimester was significantly decreased, with a count of 81.2 +/- 15.0%. 3. In the other periods, PS and PC were slightly decreased but not significantly. There was no remarkable change in PS or PC in normal third trimester pregnancy or in toxemia of pregnancy. 4. Prothrombin was increased during pregnancy, but not significantly. 5. There was a good correlation between PS and PC concentrations (r = 0.52, p less than 0.01). PMID- 2976077 TI - The antigen-major histocompatibility complex-T cell receptor interaction. A structural analysis. AB - In summary, we wish to propose that regions on MHC molecules interact with complementary regions on processed peptide antigens, and that the resultant Ag MHC complex forms a conformation with separate functional regions that are able to interact with similarly complementary areas on T cell receptors. It is the product of these interactions that determines whether a given peptide Ag is capable of binding the MHC molecule, and whether a given Ag-MHC complex is capable of stimulating a particular T cell. As more becomes known about the molecular aspects of MHC-restricted, Ag-specific T cell activation, it will become clear which amino acid residues on the Ag, MHC, and TCR are involved in these interactions. PMID- 2976079 TI - Acute intervention in myocardial infarction--role of thrombolytic therapy and adjuvant angioplasty. PMID- 2976080 TI - Neuropsychological and neuroradiological correlates in Huntington's disease. AB - Measurements of cortical and subcortical atrophy were made on CT scans of 34 patients with Huntington's disease. Significant correlations were found between the bicaudate ratio (BCR) and an eye movement scale (r = 0.44, p less than 0.01), and activities of daily living scale (r = 0.57, p less than 0.001) and the Mini Mental State Exam (r = 0.49, p less than 0.01). No correlations were found between BCR values and severity of chorea or voluntary motor impairment. A detailed neuropsychological evaluation of 18 Huntington's disease patients showed significant correlations between the BCR and Symbol Digit Modalities test (r = 0.65, p less than 0.01), and parts A (r = 0.72, p less than 0.001) and B (r = 0.80, p less than 0.0001) of the Trail Making Test. These data support work in primates that demonstrates the role of the caudate nucleus in cognitive and oculomotor functions, but not in motor control (which is governed by putamino subthalamic systems). The specific cognitive skills correlated with caudate atrophy in Huntington's disease are those reported in primate work to be served by the frontal-caudate loop system: eye movements, conceptual tracking, set shifting and psychomotor speed. PMID- 2976081 TI - Imaging of cerebral blood flow markers in Huntington's disease using single photon emission computed tomography. AB - Single photon emission computed tomography (SPECT) imaging of six Huntington's disease patients revealed a striking reduction in regional uptake of cerebral blood flow markers in vivo. Similar changes were found in one patient with "early stage" disease. The findings are compared with parallel magnetic resonance imaging (MRI) studies, and in one case, results of postmortem examination. PMID- 2976082 TI - Peripheral blood lymphocyte phenotype and function in multiple sclerosis. AB - T suppressor cell function and phenotype are abnormal in patients with multiple sclerosis, especially during the chronic progressive phase but the sub populations defined by mitogen stimulation and serological methods may not be identical. In this study, involving 45 patients with multiple sclerosis and 33 controls, there was no correlation between T suppressor function and CD8 cell phenotype in patients with multiple sclerosis or in controls. These phenotypic and functional studies cannot therefore be used interchangeably in the assessment of patients with multiple sclerosis since they provide different information about lymphocyte subpopulations. PMID- 2976083 TI - Premature aging changes in neuromuscular junctions of transgenic mice with an extra human CuZnSOD gene: a model for tongue pathology in Down's syndrome. AB - We examined the tongue muscles in several strains of transgenic mice carrying the human Zn-Cu superoxide dismutase (CuZnSOD) gene. The presence of the extra gene was confirmed in mated progeny and the gene product activity was measured in the tongue and found to be much higher than in normal littermate controls. Using electron microscopic morphometry, the neuromuscular junctions of the transgenic mice showed significant changes resembling excessive aging, with atrophy, degeneration, withdrawal, and sometimes destruction of the terminal axons, as well as the development of multiple small terminals. The myofibers showed little change except for slight hypertrophy and an increased variability in size. They also had more megamitochondria, fat droplets and lipofuscin bodies. Excess CuZnSOD generates H2O2 and hydroxyl radicals which affect both NMJ membranes and plasticity, and which may produce premature aging. The findings resemble those observed in tongues of patients with Down's syndrome, in whom an extra CuZnSOD gene is present as part of the trisomy of chromosome 21. PMID- 2976084 TI - Photodynamic therapy is potentiated by Co60 and intratumoral injection of hematoporphyrin derivative. AB - Hematoporphyrin derivative injected directly into a subcutaneous rat glioma resulted in a significant greater tumor growth inhibition than hematoporphyrin derivative injected parenterally upon stimulation by light, Co60 or by combination of the above. The photodynamic effect was analyzed in an in vivo and in an in vitro clonogenic assay. The directly injected hematoporphyrin derivative without external activation inhibited tumor growth to 34%. Light and directly injected HPD inhibited tumor growth to 3%, whereas 4, 8 and 16 Gy of Co60 produced a growth inhibition to 11%, 0.05% and 0.00% respectively. This cytotoxic effect of the ionizing radiation is further potentiated by the addition of light, resulting in a growth inhibition to 0.1%, 0.00% and 0.00% respectively for the three corresponding radiation doses. The direct intratumoral injection of HPD minimizes the side effects and increases the effect of the photodynamic therapy as compared to the parenterally administered HPD. This direct injection modality could be of potential value in the treatment of human gliomas or other tumors. PMID- 2976085 TI - Long-term follow-up of nonoperatively treated thoracolumbar spine fractures. AB - Little information is available regarding the long-term results of nonoperative treatment for fractures of the thoracolumbar spine. One thousand six hundred ninety-one fractures of the spine seen at the University of Iowa from 1935 to 1975 were reviewed; 83 fractures met strict inclusion criteria of fractures involving T10-L5. In all cases, nonoperative treatment was the only treatment received. 42 patients (51 per cent) were contacted and completed questionnaires. Twenty (48%) of these 42 patients also returned to University Hospital for a complete physical examination as well as anteroposterior and lateral flexion extension radiographs. The average time from injury to follow-up was 20.2 years, (range 11-55 years). The average age at follow-up was 43 years (range 28-70). There were 31 men and 11 women. Seventy-one percent of the injuries were the result of motor vehicle accidents. The most common sites of injury were T12-L2, which accounted for 64% of the injuries; 78% of the patients had no neurologic deficits at the time of injury. At follow-up, the average back pain score was 3.5, (0 = no pain at all, and 10 = very severe pain). No patient demonstrated a decrease in neurologic status at follow-up, and no patient required narcotic medication for pain control. Eighty-eight percent of the patients were able to work at their usual level of activity. Follow-up radiographs revealed an average kyphosis angel of 26.4 degrees in flexion and 16.8 degrees in extension. The degree of kyphosis did not correlate with pain or function parameters in the 20 examined patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976086 TI - [Transmitral flow patterns during acute coronary artery occlusion: experimental and clinical evaluations by pulsed Doppler echocardiography]. AB - To evaluate left ventricular diastolic properties during acute myocardial ischemia, transmitral flow patterns were analyzed by two-dimensional pulsed Doppler echocardiography. In a preliminary study of transient coronary artery occlusion in anesthetized open-chest dogs, the following results were obtained: decreased peak velocity of rapid filling, prolonged rapid filling period, and compensatory increase in the peak velocity of atrial filling. These changes occurred nearly concomitantly with decreased cardiac output, elevated left atrial and left ventricular end-diastolic pressures, and decreased left ventricular systolic pressures. With reperfusion of the coronary artery, these parameters rapidly returned to the base line values. In acute and transient myocardial ischemia induced by intracoronary balloon inflation during therapeutic percutaneous transluminal coronary angioplasty (PTCA) in man, similar changes in these parameters were observed, and the changes were more marked in PTCA for the left anterior descending artery as compared with that of the right coronary artery. In conclusion, noninvasive two-dimensional pulsed Doppler echocardiography is of practical use in evaluating deterioration of the diastolic function of the left ventricle during acute and transient myocardial ischemia. PMID- 2976087 TI - [Left ventricular diastolic properties before and after percutaneous transluminal coronary angioplasty evaluated by pulsed Doppler echocardiography]. AB - Left ventricular diastolic properties were assessed by pulsed Doppler echocardiography before and after percutaneous transluminal coronary angioplasty (PTCA) in 30 patients. Four parameters were measured from records obtained by directing the ultrasonic beam from the cardiac apex to the base and by locating the sample volume at the mitral orifice: 1) the peak velocity of the rapid filling wave (R), 2) the peak velocity of the atrial contraction wave (A), 3) the acceleration half time (Ta: the time from half of the peak velocity to the peak velocity of the R, and 4) deceleration half time (Td: the time from the peak velocity to half of the peak velocity of the R). The ratio of the peak velocity of the atrial contraction to that of the rapid filling wave (A/R) was also calculated. 1. The R increased significantly after PTCA (p less than 0.01), whereas no significant change was observed in the A. 2. The A/R decreased significantly after PTCA (p less than 0.01). 3. PTCA did not influence significantly the acceleration half time or the deceleration half time. The augmented rapid filling observed after PTCA may reflect improvement of left ventricular myocardial relaxation. Our method sensitive in assessing diastolic properties is helpful for evaluating left ventricular function after PTCA. PMID- 2976088 TI - [Myocardial perfusion assessed by dynamic computed tomography before and after percutaneous transluminal coronary angioplasty]. AB - A method for quantitatively and visually assessing myocardial perfusion using a new transmission computed tomography scanner and contrast media injection was devised. Myocardial perfusion was assessed before and after percutaneous transluminal coronary angioplasty (PTCA). Six patients with left anterior descending coronary artery stenosis undergoing PTCA were successfully studied. Dynamic scans, each with a scan time of one sec, were performed at the mid left ventricular level using bolus injections of contrast medium from the inferior vena cava. Regions of interest (ROIs) were located in the left ventricular cavity and in the myocardium perfused by the left anterior descending coronary artery, and gamma-variate fitted time density curves were recorded within the first pass phase, excluding recirculation. Cardiac outputs calculated from the ventricular curves of the ROIs in the left ventricular cavity correlated well with the thermodilution measurements using Swan-Ganz catheters (r = 0.90, p less than 0.01). The ratios of myocardial blood flow (F) to the tissue element volume (V) were calculated (F/V) from the myocardial curves of the ROIs in the myocardium as parameters of myocardial perfusion, according to the Zierler's principle. The F/Vs in the myocardium perfused by the left anterior descending coronary artery before PTCA were significantly less than those of the control subjects (4.4 x 10( 2) +/- 1.2 x 10(-2) vs 8.1 x 10(-2) +/- 2.9 x 10(-2): p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976089 TI - Immunopathogenesis of HIV infection. AB - HIV may cause CD4-positive T cell depletion not only by direct infection but by causing damage to normal maintenance mechanisms. Findings in support of such a concept are presented. Results suggesting that immune reactions against HIV may contain elements of a negative nature are presented, in particular in relation to viral enhancement by antibodies. A question is also raised whether HIV replication has been proved to be reduced in a significant manner by immune reactions. PMID- 2976090 TI - Potentiation of the antitumor effect of 5-fluoro-2'-deoxyuridine esters in combination with acyclothymidine esters on L1210 in mice via oral administration. AB - Fifteen pyrimidine-related compounds were evaluated for their ability to inhibit enzymatic degradation of 5-fluoro-2'-deoxyuridine (FUdR). Acyclothymidine [5 methyl-1-(2'-hydroxyethoxymethyl)uracil] showed the highest inhibitory effect on the phosphorolytic degradation of FUdR in various tissue homogenates derived from mouse, rat, and beagle organs. Both the drug (FUdR) and the inhibitor (acyclothymidine) were esterified with appropriate aliphatic acids in order to synchronize their behavior after simultaneous oral administration. The antitumor activity of orally administered FUdR esters was potentiated by the simultaneous oral administration of the acyclothymidine esters, but not by acyclothymidine. PMID- 2976091 TI - Application of a stereospecific high-performance liquid chromatography assay to a pharmacokinetic study of etodolac enantiomers in humans. AB - An HPLC assay suitable for pharmacokinetic analysis of enantiomers of etodolac [(+/-)-1,8-diethyl-1,3,4,9-tetrahydropyrano[3,4-b] indole-1-acetic acid] was developed. Following addition of internal standard (IS), (+/-)-2-(4 benzoylphenyl)butyric acid, the constituents were extracted from the specimen into a mixture of isooctane:isopropanol (95:5). The organic layer was evaporated and the drug and IS were sequentially derivatized with ethyl chloroformate and iota(-)-alpha-phenylethylamine. The diastereoisomers thus formed were extracted and chromatographed on a normal-phase column, with a mobile phase consisting of hexane:ethyl acetate:isopropanol (85:15:0.2) at a flow rate of 2 mL/min. The etodolac diastereoisomers were separated with a resolution factor of 6.4 and detected at a wavelength of 280 nm. Excellent linear relationships were found between the peak area ratios (etodolac:IS) and the plasma and urine concentrations (0.2-20 mg/L), with intra- and interday variations of less than 10.1%. The assay was applied to a preliminary pharmacokinetic study following seven repeated oral administrations of 200 mg/12 h of racemic etodolac to two healthy subjects. The plasma concentrations of the active S-(+)-enantiomer were considerably less than those of the inactive antipode (AUC S:R, 2.5:30.9 mg.L-1.h 1) due to a greater volume of distribution of the latter (S, 101 and 135 L versus R, 24 and 17 L). Considerable concentrations of conjugated enantiomers were also found in plasma (AUC conjugated: intact: S, 1.1; R, 0.23). PMID- 2976092 TI - A new criterion for selection of pharmacokinetic multiexponential equations. AB - In linear pharmacokinetics, the time course of the plasma concentration of a drug, Ct, is expressed by the sum of exponential terms, (formula; see text) This article proposes a new statistical criterion for discriminating between alternate polyexponential models. According to this new criterion, the model that best interprets a set of experimental data points is that which minimizes the area between the approximate confidence limits of Ct. PMID- 2976093 TI - Pathophysiologic basis of canine muscle disorders. PMID- 2976094 TI - Carbon dioxide laser treatment of acne rosacea and rhinophyma: how I do it. PMID- 2976095 TI - [Coronary artery and left ventricle angiography--diagnostic and therapeutic application]. PMID- 2976096 TI - [The kidney and adrenal gland angiography--their therapeutic application in embolization and angioplasty]. PMID- 2976097 TI - [Angiography of the upper and lower limbs]. PMID- 2976098 TI - [Usefulness of rest-redistribution thallium scan for the indication of PTCA in an interesting case with acute myocardial infarction]. PMID- 2976099 TI - [Aorto-coronary bypass grafting in a patient with a single coronary artery complicated with myocardial infarction]. PMID- 2976100 TI - [Coronary artery angioplasty in 239 patients]. PMID- 2976101 TI - [Disorganization of myocardial fibers. III. Effect of myocardial hypertrophy on electrocardiographic criteria]. PMID- 2976102 TI - [Cholecystectomy through a small incision in the abdominal wall]. PMID- 2976103 TI - [Ultrastructural analysis of atrial cardiomyocytes of rats exposed to acceleration +5Gz]. AB - Ultrastructural changes in atrial cardiomyocytes of rats chronically exposed to +5 Gz were investigated. Immediately after exposure most cardiomyocytes showed hypertrophic and hyperplastic changes in practically all cell organelles. During a month-long recovery destructive changes that seemed to reflect hypertrophy regression did not return to norm. The above observations concerning changes in the system of atrial hormones together with pertinent data in the literature suggest that a factor produced by the system may be involved in the complex mechanism of acceleration-induced effects on the animal body. PMID- 2976104 TI - Myosin ATPase activity during avian cardiac and skeletal muscle development. AB - The myosin ATPase activity and myosin light chain composition in developing chick heart and skeletal muscles were studied and compared. Embryonic myosin was purified and characterized from day 7 to day 19 of embryogenesis. Embryonic cardiac myosin generally showed the same Ca2+-activated myosin ATPase activity level as the adult value. In comparison, pooled pectoralis and hindlimb skeletal muscles from day 10 through day 19 showed myosin ATPase activities that were all significantly less than the adult counterpart. The myosin light chain pattern of embryonic cardiac myosin remained relatively constant like the myosin ATPase activity, whereas developmental changes were observed in skeletal myosin light chains. PMID- 2976105 TI - [Laparoscopy in ascites]. PMID- 2976106 TI - Analgesic activity of two synthetic immunomodulators, muramyl dipeptide and adamantylamide dipeptide in mice and rats. AB - The potency of two synthetic immunomodulators, muramyl dipeptide and adamantylamide dipeptide, which have the immunoadjuvant and immunomodulatory activity on pain threshold was studied. Two different analgesiometric procedures were employed: hot plate test and acetic acid writhing test in mice and rats. Both compounds were injected intravenously (1-4 mg/kg), intraperitoneally (5-50 mg/kg) and intracerebroventricularly (0.5-4 mg/kg) and were able to produce mild transient analgesia in both species. Writhing response was more influenced after systemic administration of drugs while hot plate latencies was not. On the contrary, latencies in hot plate test were more affected than the writhing response after intracerebroventricular administration. Dose response curve showed a bell shaped feature typical for peptides. Pretreatment with naltrexone, an opiate antagonist, did not prevent the analgesic action of tested compounds. The hyperalgesia induced by administration of parachlorophenylalanine, a serotonin depletor, could be prevented by administration of a nonanalgesic dose of MDP (0.025 mg/kg). At higher dosages (1 mg/kg) MDP was able to antagonize also general toxic effects of pCPA. These results support the possibility of participation of central serotonergic structures in MDP and AdDP induced analgesia. The peripheral mechanism of action, however, can not be completely ruled out. PMID- 2976107 TI - [Clodronate]. PMID- 2976108 TI - [The majority of suppressor T lymphocytes (CD4- CD8+ CD11+) expresses S-100 protein]. PMID- 2976109 TI - [Identification of dopamine-D2 receptors on lymphocytes using the binding technic]. PMID- 2976110 TI - [Hemodynamic, renal and humoral effects of the infusion of human natriuretic peptide in hypertensive patients]. PMID- 2976111 TI - [In vitro culture of humana trophoblastic tissue: evaluation of its functionality in basic conditions and after administration of potentially toxic substances]. PMID- 2976112 TI - Chitinase from Neurospora crassa. PMID- 2976113 TI - Arthus phenomenon. PMID- 2976114 TI - Alternative pathway of complement. PMID- 2976115 TI - An examination of the association between vertebral deformities, physical disabilities and psychosocial problems. AB - In order to measure the clinical consequences of spinal osteoporosis, we correlated a number of physical and psychosocial dysfunctions with the degree of vertebral deformity in 204 women aged 55-75 yr. We employed 5 standardized questionnaire instruments: the physical dimension of the Sickness Impact Profile (SIP), an analog back pain scale, a 24-question instrument eliciting back-related disabilities, and assessments of self-esteem and mastery. Using these results as the dependent variables, we performed 5 separate stepwise regression analyses; independent variables were vertebral deformity score and 11 possible confounders. Using Pearson correlation coefficients, we found a high degree of correlation between each of the 5 primary outcome variables. Vertebral deformity score played a small but statistically significant role in the 3 models whose dependent variable was related to physical dysfunction, but it accounted for only 4-10% of the variance of these models. Mastery and self-esteem were related to overall poor health and attendant physical disabilities but not to vertebral deformity score. In this selected cohort of women, the occurrence of mild to moderate vertebral deformities caused little loss of physical function or function of psychosocial problems because of vertebral deformity. Although we were able to demonstrate a statistically significant handicap related to the total vertebral deformity score, several other clinical variables appeared to have a greater impact. PMID- 2976116 TI - Dehydroepiandrosterone sulphate and dehydroepiandrosterone in serum: differences related to age and sex. AB - Serum concentrations of dehydroepiandrosterone sulphate (DHAS) were determined in 590 healthy women aged 20-87 yr. Simultaneous assays of dehydroepiandrosterone (DHA) were performed in 417 of the women. DHA and DHAS levels correlated negatively with age while the DHA/DHAS ratio proved to be unrelated to age. When values for 60 healthy men in the age range 20-84 yr were compared with those obtained in 60 randomly-selected healthy women who were exactly age-matched, the DHAS levels were found to be significantly lower and the DHA/DHAS ratios significantly higher in the women. These results might be of use in establishing normal clinical ranges for serum DHA, DHAS and the DHA/DHAS ratio in women. PMID- 2976117 TI - The effect of medical education on the attitudes of medical students towards disabled people. AB - The negative attitudes of society towards disability and the resulting prejudicial behaviour affects the lives of disabled people. One of the declared aims of the Faculty of Medicine of the University of Southampton is to improve attitudes. Hence, an attempt has been made to assess differences in the attitudes held by first- and fourth-year medical students, senior house officers and members of the general public towards disabled people. The measurement instrument used was the 'Attitudes Towards Disabled Persons (ATDP) Scale'. The total number of subjects in the survey was 428, of whom 263 (61%) responded. No significant differences were found between the attitudes of the four groups studied. However, a subpopulation of subjects who agreed with the statement that 'Disabled people cause more problems to doctors than non-disabled people' had attitudes which were significantly more negative. PMID- 2976119 TI - [Histological findings of supernumerary teeth in a case of cleidocranial dysostosis]. PMID- 2976120 TI - Further purification and characterization of hepatopoietin. PMID- 2976118 TI - One-step gene disruption by cotransformation to isolate double auxotrophs in Candida albicans. AB - The Candida albicans LEU2 gene was disrupted by substituting lambda DNA for a small deletion within the LEU2 gene. Cotransformation with a selectable URA3 ARS vector was used to introduce a linear fragment containing the disruption into the genome of a C. albicans ura3 deletion mutant. Cotransformants containing the lambda DNA were identified by colony hybridization and the URA3 plasmid was subsequently cured. Leu2 disrupted heterozygotes were detected by Southern hybridization and one disruptant was subsequently treated with UV irradiation. Only one leu2 ura3 mutant (SGY-484) was isolated out of 11,000 mutagenized cells. SGY-484 was transformed to Leu+ with either the C. albicans or Saccharomyces cerevisiae LEU2 gene. Southern hybridization analysis revealed that the mutant is not homozygous for the disruption; the leu2 mutation reverts and is most likely a point mutation. Unexpectedly, an ade2 ura3 mutant was isolated from the same mutagenesis. PMID- 2976121 TI - In vitro effect of PTH on normal T cell functions. AB - Uremia is associated with impairment of various cell-mediated immunity functions. The effect of parathyroid hormone (PTH) - known to be elevated in uremia - on several T cell functions has been studied. Normal peripheral blood lymphocytes incubated with increasing amounts of human PTH (HPTH) or bovine PTH (BPTH) showed a considerable decrease (up to 40%) in lectin-induced lymphocytes transformation, significant decrease in helpers to suppressors ratio, and marked inhibition of E rosette formation and T11-positive cells. PTH alone showed no cytotoxic effect on lymphocytes when incubated with or without mitogens. Glucagon, in concentrations up to 10-fold those found on uremia, had no effect on T cell function. Thus the effect of PTH was specific to the hormone action. The direct effect of PTH on normal T lymphocytes and some of their immunological responses is not clear. However, the results of this study support the hypothesis that excess blood levels of PTH may play a role in the pathogenesis of the impairment of the immune response in uremia. PMID- 2976122 TI - Plasma levels of human atrial natriuretic factor in patients treated by hemodialysis and continuous ambulatory peritoneal dialysis. AB - We measured plasma levels of immunoreactive human atrial natriuretic factor (ANF) in chronic renal failure patients treated by hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD). Predialysis plasma ANF was significantly higher in HD patients (271.8 +/- 173.4 pg/ml) as compared to CAPD patients (81.8 +/- 80.5 pg/ml) and healthy subjects (31.5 +/- 19.8 pg/ml). Plasma volume was higher in HD patients than in CAPD patients. Plasma ANF and plasma volume showed a significant positive correlation. In HD patients, high plasma ANF value decreased significantly to a value comparable with that of CAPD patients after each dialysis. The removal rates of ANF by HD and CAPD were comparable. Ultrafiltration corresponding to 2% of body weight without dialysis also reduced plasma ANF. Thus, the difference in plasma ANF values between HD and CAPD patients seems to be mostly due to the difference in plasma volume, indicating that plasma ANF is sensitive to volume status even in chronic dialysis patients. PMID- 2976123 TI - Effects of insulin on rat brain noradrenaline. AB - A single intracardial injection of streptozotocin produced a significant increase in rat hypothalamic noradrenaline while no changes were observed in the olfactory tubercles. The parenteral administration of a single dose of insulin decreased rat hypothalamic noradrenaline; the effect had a rapid onset and lasted for at least six hours. Similar noradrenaline reductions were observed in the olfactory tubercles but in this tissue the depletion started later and recovered earlier. In addition, in olfactory tubercles after insulin injection, tyrosine level and dopamine metabolism were increased. The results show that the increases in hypothalamic NA observed in streptozotocin diabetic rats are counteracted by insulin administration and possibly the consequence of changes in noradrenaline turnover. PMID- 2976124 TI - [Spinal intramedullary tumor with exophytic growth]. AB - We experienced 22 cases of spinal intramedullary tumor, in which 5 cases were accompanied with exophytic growth. We discussed their clinical and radiological features, and therapeutic problems. Concerning the locations of tumors with exophytic growth, the most common site was conus medullaris, accounting for 60% (3 cases). Histopathological findings were astrocytoma in two cases, and in one case, mixed glioma, ependymoma and hemangioblastoma. In neurological observation, the most common initial symptom was back pain and lumbago, suggesting root pain. No neurological features distinguishable from those of extramedullary tumors were presented. In radiological examination, myelography and CT myelography were very helpful for diagnosis. Myelography and CT myelography showed extramedullary mass, shift and deformity of spinal cord that was not serious as compared with the size of extramedullary mass, and showed the portion where the spinal cord was swollen. Good outcomes were obtained in a case with total removal, and two cases with subtotal and partial removal that were managed with additional irradiation and chemotherapy. However recurrence and intracranial seeding made prognosis poor in two cases where total removal was impossible. We thought that postoperative careful follow-up was necessary not only to detect recurrence but also to detect intracranial seeding in the cases of spinal intramedullary tumor with exophytic growth. PMID- 2976125 TI - Hematoporphyrin uptake in atherosclerotic plaques: therapeutic potentials. AB - Atherosclerotic plaques were induced in abdominal aortas of rabbits. At 8 weeks, 5 mg of dihematoporphyrin ether (Photofrin II) per kg was injected intravenously followed by sacrifice of the animal, fluorescence microscopy, and quantitative assay of porphyrin in the plaque-containing aortas at 1, 12, 24, 48, and 72 hours. Photofrin II was taken up preferentially by the plaque, with the highest plaque to normal wall ratio occurring at 48 hours. Phototherapy was carried out in 13 animals in each of which two plaques had been induced. With a 630-nm light source 48 hours after the infusion of Photofrin, one of the pair of plaques was treated while the other served as a control. Animals were killed at 2, 4, and 6 weeks. The 6-week specimens showed the most dramatic reduction in plaque in comparison to controls. Photodynamic therapy may provide an alternate strategy in dealing with focal atherosclerosis. PMID- 2976126 TI - Modification of experimental post-subarachnoid hemorrhage vasculopathy with intracisternal plasmin. AB - This study using pigs was designed to determine the efficacy of delayed subarachnoid plasmin injection in preventing the vasculopathy secondary to experimental subarachnoid hemorrhage. Animals received cisterna magna injections of either plasmin or saline 2, 4, or 6 days after double subarachnoid blood injection. The cerebral blood vessels of all animals were examined histologically for evidence of intimal proliferation and medial necrosis. Angiography was not done. The results indicate a progressive increase in the extent and severity of intimal proliferation the longer plasmin injection is delayed. In contrast, the severity of medial necrosis did not correlate with the delay. We conclude that subarachnoid clot lysis is an effective way to prevent vascular injury from subarachnoid blood and that, the sooner it is done after hemorrhage, the more effective it will be. PMID- 2976127 TI - [Morphological changes in the rat brain after intracardiac and intraperitoneal administration of quinolinic acid]. PMID- 2976128 TI - Increased plasma levels of atrial natriuretic peptide in patients with chronic renal failure: effect of noradrenaline infusion. AB - Basal plasma atrial natriuretic peptide (ANP) and blood pressure were measured in 11 patients with chronic renal failure before requirement of dialysis, 13 patients on chronic dialysis, and 28 control subjects (Study 1). Changes in ANP during noradrenaline infusion were determined in eight patients with chronic renal failure before dialysis, 12 patients on chronic dialysis, and 17 control subjects (Study 2). ANP was also measured in 14 healthy control subjects during angiotensin II infusion (Study 3). Study 1 showed a significantly greater ANP in patients before the stage of dialysis (median 23 pg/ml) and in dialysis patients (34 pg/ml) than in control subjects (19 pg/ml) P less than 0.01 for both. In Study 2, noradrenaline induced an increase in ANP in the non-dialysed patients (P less than 0.05) and in the control subjects (P less than 0.01), but not in the dialysis patients. According to Study 3, ANP was unchanged during angiotensin II infusion. Blood pressure was increased in all groups during noradrenaline and angiotensin II infusions. It can be concluded that ANP Is increased both in patients with chronic renal failure before requirement of dialysis and in patients on maintenance dialysis. It is suggested that noradrenaline stimulates ANP release. PMID- 2976129 TI - Tetrahydroaminoacridine potentiates neurotoxicity of quinolinic acid in rat striatum. AB - Intraperitoneal injection of 5 mg/kg of tetrahydro-9-amino-acridine (THA) in rats 1 h before intrastriatal injection of 50-150 nmol of quinolinic acid potentiated the local neurotoxicity as indicated by measurements of striatal levels of glutamate decarboxylase and choline acetyltransferase. A larger dose (10 mg/kg) THA had no significant effect. The results are discussed in terms of THA's binding to various elements of the NMDA/sigma receptor complex and reported data on such binding are confirmed. PMID- 2976130 TI - Dextromethorphan does not protect against quinolinic acid neurotoxicity in rat striatum. AB - Dextromethorphan (DM, 40 or 80 mg/kg, i.p.) and MK-801 (3 or 10 mg/kg, i.p.) were compared in their ability to prevent the depletion of choline acetyltransferase (ChAT) activity in the rat striatum following intrastriatal injection of quinolinic acid. DM did not reduce striatal ChAT depletion following injection of either 300 or 150 nmol of quinolinic acid. Following injection of 300 nmol of quinolinic acid, MK-801 significantly reduced striatal ChAT depletion at a dose of 3 mg/kg and completely prevented striatal ChAT depletion at a dose of 10 mg/kg. In contrast to the potent neuroprotective action of MK-801, DM does not protect striatal cholinergic neurons from an acute challenge by an NMDA receptor agonist. PMID- 2976131 TI - Pharmacological manipulation with the descending serotonergic system or transection of the mouse spinal cord has no effect on ependymal ultrastructure. AB - In order to investigate an effect of descending nerve fibres on mouse spinal cord ependymal ultrastructure, pharmacological manipulation with the serotonergic system or transection of the spinal cord was done. Biochemical analysis showed an 83% reduction of serotonin content in spinal cord tissue after p chlorophenylalanine injections and a 93% reduction after transection. However, none of the experimental animals showed changes in ependymal ultrastructure compared to control animals as revealed by electron microscopy. PMID- 2976132 TI - In vivo and in vitro autoradiographic labelling of central dopaminergic systems with [3H]GBR12783 in rodents. AB - Labelling of central dopaminergic systems with the dopamine uptake inhibitor [3H]GBR12783 was performed in vivo in mice by i.v. injection of a tracer dose of the ligand (40 microCi) 1 h before sacrifice. Autoradiographic pictures obtained from these experiments were quantified. The data were compared to those obtained by in vitro labelling of rat brain slices with this ligand. Significant specific labelling was observed both in vivo and in vitro in the striatum, the nucleus accumbens, the olfactory tubercle and the substantia nigra. PMID- 2976133 TI - Combined percutaneous aortic valvuloplasty and percutaneous coronary angioplasty. PMID- 2976134 TI - Calcifying aponeurotic fibroma. PMID- 2976135 TI - Workers Comp reimbursement is available for NP services. PMID- 2976136 TI - A novel two-site immunoradiometric assay for beta-endorphin using nitrocellulose as solid phase. AB - A two-site immunoradiometric assay for the highly specific direct quantitation of nonacetylated beta h-EP in crude brain tissue samples has been developed with a detection limit of 10 fmol per well. The assay used two different antibodies with distinct specificities: a polyclonal rabbit anti-beta h-EP antibody binding between the middle portion and the C-terminal end of beta h-EP was bound to nitrocellulose membrane discs, a solid phase with a high protein binding capacity. In the following two incubation steps, the beta h-EP containing crude tissue extract--or the beta h-EP-standard--and, subsequently, the 125I-labeled monoclonal 3-E7 mouse antibody directed against the N-terminus of beta h-EP were added. Binding of beta h-EP to the solid phase antibody in the first incubation step was not affected by the addition of cross reacting opioid peptides derived from beta h-LPH up to 10 pmol per disc. Nonspecific binding of the labeled antibody to the solid phase could be lowered to 3% of total counts by the use of PBS containing nonfat dry milk as blocking solution and incubation buffer, a procedure that did not reduce maximum specific binding. Dilution studies performed with extracts sampled from the anterior hypothalamus excluded the interference of tissue factors in the assay. PMID- 2976137 TI - [Fixation of screws in spongy bones using plastic pins]. PMID- 2976138 TI - [Transportation devices in the system of rehabilitation of patients and invalids]. PMID- 2976139 TI - [The centenary of the Department of Traumatology and Orthopedics of the S.M. Kirov Order of Lenin, Red Banner, Military Medical Academy]. PMID- 2976140 TI - cDNA clones from autocrine thymic lymphoma cells encode two mitogenic proteins, a serine protease and a truncated T-cell receptor beta-chain. AB - Cell lines derived from primary X-ray induced T cell lymphomas (PXTL) of C57BL/6 mice secrete into the medium factor(s) required for their growth. These autocrine factor(s) are distinct from previously described growth factors. cDNA cloning experiments were performed in an attempt to identify these autocrine factor(s). cDNA clones were selected by mRNA size, differential expression, and mitogenic activity of their translation products (Xenopus expression system) on PXTL cells. Two different cDNA clones yielded distinct mitogenic proteins. One clone encodes an altered form of the T cell receptor beta-chain which is truncated at the N terminus to amino acid 49 of the constant region beta 2. The second clone encodes a serine protease which is identical to factor H or granzyme A from cytotoxic T cells. The 5' portion of the cDNA encoding the serine protease derived from PXTL cells differs from that derived from cytotoxic T cells. This difference results in distinct signal peptides. Unlike cytotoxic T cells, PXTL cells do not store the serine protease intracellularly but secrete it. PMID- 2976141 TI - Negative regulation of c-myc transcription involves myc family proteins. AB - Expression of the c-myc gene is suppressed in NIH 3T3 mouse fibroblast cells infected with recombinant retroviruses expressing high levels of v-myc (10-fold greater than those of c-myc). Suppression of steady state levels of c-myc mRNA occurred at least in part at the level of transcription from c-myc promoters P1 and P2, and involved v-myc protein since cells infected with constructs containing frameshifts and deletions in v-myc had normal levels of c-myc mRNA and protein. Suppression of c-myc expression was also observed in fibroblasts transfected with a N-myc expression vector and in fibroblasts infected with a c myc retrovirus. These findings establish that v-myc protein is involved either directly or indirectly in a regulatory circuit which represses c-myc proto oncogene transcription. Feedback regulation of c-myc transcription may be relevant in establishing the lineage specific expression of myc family proto oncogenes. Reduced steady state levels of c-myc mRNA were also observed in NIH 3T3 cells infected with 12S and 13S EIA recombinant retroviruses suggesting that the exogenous oncogene of adenovirus, EIA, can alleviate the requirement of myc for cell growth and may also share transcriptional target genes. PMID- 2976143 TI - Life events in neurological patients with headache and low back pain (in relation to diagnosis and persistence of pain) by Jorgen Jensen. PMID- 2976142 TI - MMPI changes following behavioral treatment of chronic low back pain. AB - A large sample of patients with chronic low back pain were studied on admission to a behaviorally oriented in-patient pain program, at program completion, and at 1 month follow-up. Cluster analysis of admissions MMPI scores were used to identify 4 patient subgroups each for males and females. There were no subgroup differences found on any of the admissions demographic, pain report, or physical function measures, or differential treatment outcome based on subgroup. All groups began with high levels of pain and disability, yet improved dramatically following treatment. There was a general normalization of the MMPI reflected by elevated MMPI scales found at admission showing significant decreases at follow up testing. Subgroups derived from follow-up MMPI testing were related to physical functioning at follow-up and pain report measures, with the elevated subgroups showing higher levels of continued pain and disability. PMID- 2976144 TI - [Ectoparasites and phoresants of the root vole (Microtus oeconomus Pall.) in the Polesye]. AB - Drainage melioration in the Polesye resulted in a sharp increase in the number of tundra vole (Microtus oeconomus Pall.) which quickly included into the parasitocenosis of the drained land. There were found 26 parasitic and nonparasitic species of Gamasoidea, 3 species of Ixodidae and 2 species of Trombiculidae, 10 species of Aphaniptera, 3 species of Anoplura but there was found no species specific only for the tundra vole. All found parasites occur on many hosts. This explains wide parasitic links of the tundra vole with other homothermic animals which especially extensive with the bank vole and it may have important consequences for epizootiology of tularemia and tick-borne encephalitis. PMID- 2976146 TI - [Effect of cattle color, age, size and behavior on the intensity of the attack and sucking attachment by gadflies]. AB - Mechanisms of effect of some morphophysiological parameters of cattle (age, colour, weight, skin area, intensity of defensive movements) on the attacking and attaching activity of tabanid flies were studied. Investigations were carried out in the south of Pskov Province in small herds by the method of simultaneous recording of attacking and attaching tabanids on all animals of the herd. It was established that differences in the attaching activity of Haematopota, Tabanus and Hybomitra flies depend mainly on the parameters affecting the efficiency of their attacks and connected with the age of animals, intensity of defensive movements and the host's skin area. The former correlates with the age of cows negatively while the latter positively. Therefore, more tabanids attach themselves to old animals than to young ones. For Chysops flies the main factor determining their intensity of attachment is the intensity of attacking. When attacking the Chrysops flies show preference to animals of dark colour independent of their age. It is shown that the increase in the attacking intensity results in the decrease of its efficiency and therefore reduces the probability of attachment for each individual. PMID- 2976145 TI - [The incidence of toxoplasmosis among wild vertebrates of Turkmenia (based on serologic data)]. AB - 985 wild small mammals and birds were serologically investigated for toxoplasmosis from 1981 to 1984 in Turkmenia: antibodies to Toxoplasma gondii in the indirect hemagglutination and immunofluorescent complement fixation reactions according to Goldwasser and Shepard were found in 247 (25.0%), in 11 of 17 investigated species of mammals and in 13 of 22 species of birds. A sharp rise in the toxoplasmosis infection level of wild mammals was found out serologically for the first time (from 1.2% in autumn, 1982 to 72.6% in spring, 1983) followed by its reduction against the reduction in the number of animals. This must have taken place as a result of heavy toxoplasmosis epizootics in the region of investigations which is frequented by wild and domestic cats. PMID- 2976147 TI - [Faunistic analysis of the fleas on small mammals in the trans-Aral Sea area]. AB - Materials on occurrence and abundance of 36 species of fleas on 17 species of small mammals are systematized. A great similarity between the faunas of ectoparasites of rodents and predators has been shown by means of special indices. The conception of parasitic field is formulated which implies the phenomenon of community of the fauna of plague vectors parasitic on its potential carriers. Some aspects of the formation of parasitic field and its possible effect the epizootic process are considered. It is established that the similarity between the faunas of fleas from different animals is ensured in this region first of all by a wide distribution of specific parasites of gerbils. PMID- 2976148 TI - [Ctenophthalmus rettigi Rothschild, 1908 (Hystrichopsyllidae) fleas of Mesocricetus]. AB - Fleas of Ctenophthalmus rettigi Rothschild, 1908 (12 females, 12 males) collected in 1980 and 1982 in the northern Caucasus in the Levashi District of Dagestan not far from the village of Urma were referred by the authors to the nominative subspecies described from Romania. According to their taxonomic characters fleas of this species, collected in different places of Transcaucasia, are also referred to the nominative subspecies and not to C. rettigi smiti Klein, 1962, as before (Rostigaev, 1967; Tiflov, Skalon, Rostigaev, 1977 and others). The ways of distribution of hosts of C. rettigi fleas are suggested. PMID- 2976149 TI - Manipulative therapy. PMID- 2976150 TI - [The course of a streptococcal infection during treatment at a hospital and polyclinic]. PMID- 2976151 TI - [Experience of teaching in a topical lecture series on gastroenterology in childhood]. PMID- 2976152 TI - [Sex education and behavior of girls]. PMID- 2976153 TI - [Nephrologic care of children in Armenia]. PMID- 2976154 TI - [Atypical variant of the familial form of chorea in children (characteristics of its intrafamilial polymorphism)]. PMID- 2976155 TI - Cognitive differentiation, back pain, and psychogenic pain drawings. AB - Two groups of subjects with back pain were studied (n = 67). All subjects underwent a battery of psychological tests which included a test of psychological differentiation (the Rod-and-Frame Test), a test of psychological defenses (Meta contrast Technique), and the Eysenck Personality Inventory. In addition, the subjects drew a pain picture and answered a questionnaire on pain-related issues. Each group of subjects was then subdivided into 4 groups depending on the judged abnormality of their pain drawings. The two main groups were compared overall and the subgroups of the two back-pain groups compared with each other. The Meta contrast Technique results show that depression was quite common among back-pain subjects as a whole compared to a painless group of subjects. Differences were found on field-dependence; subjects with abnormal pain drawings were more field dependent than those with "normal" pain drawings. Few other between-group statistical differences were noted between the two back-pain groups. Expected scores on hysteria were not noted among the abnormal drawing makers, neither were there differences between the groups on the items of pain duration, physical functionings, or of depression. PMID- 2976156 TI - Mechanism of transcription termination in eukaryotic cell. AB - We developed an in vivo screening system for the DNA region involved in transcription termination. This system is based on the idea that the CAT gene bearing transcription terminator upstream of poly(A) signal should produce a low CAT activity in the transfected cell. Using this system, we located termination elements in the human gastrin gene and adenovirus E1 gene. Furthermore, the terminator of gastrin gene was identified by S1 nuclease mapping and in vitro transcription. PMID- 2976157 TI - Selenocysteine on glutathione peroxidase may be converted from phosphoserine on the apo-enzyme synthesized with an opal suppressor phosphoseryl-tRNA. AB - There are two possible mechanisms (co- or post-translational) for incorporation of Se into glutathione peroxidase in which selenocysteine presents at the active site of the enzyme and corresponds to UGA on the mRNA. We studied the above mechanisms using opal suppressor tRNA in mammals. Opal suppressor tRNA did not accept any selenocysteine and phosphoseryl-tRNA did not change to selenocysteyl tRNA. Meanwhile, phosphoprotein changed to a protein containing selenocysteine by the incubation with H2Se and some enzymes. From these results, we propose that phosphoserine on glutathione peroxidase (apo-enzyme), which is synthesized with phosphoseryl-tRNA, is converted to selenocysteine in the mature enzyme, by a posttranslational mechanism. Opal suppressor tRNA may play a role to synthesize the apo-enzyme of glutathione peroxidase. PMID- 2976158 TI - Detection of deoxyribonucleoside-triphosphate imbalance death-induced DNA double strand breakage in FM3A cells by orthogonal-field-alternation gel electrophoresis (OFAGE). AB - The mechanism of intracellular deoxyribonucleoside-triphosphate (dNTP) imbalance death of mouse mammary tumor FM3A cells was studied. When the cells were exposed to 5-fluorodeoxyuridine, deoxyadenosine, or 2-chlorodeoxyadenosine, dNTP pool imbalance resulted. The imbalance was followed by DNA double strand breaks and subsequent cell death. The DNA double strand breaks have been directly examined by means of orthogonal-field-alternation gel electrophoresis (OFAGE). Fragmented DNA band appeared to be approximately 100-200 kb in size. PMID- 2976159 TI - Feeding, drinking and temperature responses to intracerebroventricular beta endorphin in the domestic fowl. AB - Four experiments were conducted to evaluate the effect of beta-endorphin (beta END) on feeding, drinking and colonic temperature in rapidly growing (Rock Cornish; RC) and slow growing (Single-Comb White Leghorn; SCWL) stocks of chickens. In the first experiment RC cockerels were injected intracerebroventricularly (ICV) with 0, 1.5, 3.0 and 6.0 micrograms of beta-END. In the second experiment RC cockerels were injected ICV with 0.5, 1.0 and 2.0 micrograms of beta-END. Experiments 3 and 4 were conducted identically to Experiment 1 and 2, respectively, except SCWL were used. Administration of beta END at levels between 1.5 and 6.0 micrograms produced a significant curvilinear increase in feeding in both RC and SCWL chicks. In RC chicks, feeding was significantly elevated at 45 min and from 90 through 240 min postinjection, whereas in SCWL chicks feeding was increased from 90 through 300 min postinjection. Water intake was depressed in RC and SCWL from 60 through 90 min and from 30 through 60 min postinjection, respectively. Significant increases in water occurred at 180 and 300 min postinjection in SCWL. beta-END also induced a significant hyperthermia in RC and SCWL from 30 through 240 min and from 15 through 180 min postinjection, respectively. At low levels of beta-END, i.e., 0, 0.5, 1.0 and 2.0 micrograms, feeding, drinking and body temperature were significantly increased in both stocks. Feeding in RC chicks was stimulated in a linear fashion from 180 through 300 postinjection while feeding in SCWL was stimulated in a curvilinear manner from 180 through 240 min postinjection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976160 TI - Effects of undernutrition during suckling and of training on the hypothalamic beta-endorphin of young and adult rats. AB - The involvement of the hypothalamic beta-endorphinergic system in behavioral processes has previously been studied in adult rats. In the present report, we studied the effects of undernutrition and of inhibitory avoidance training on the hypothalamic beta-endorphin-like immunoreactivity of 21-day-old and adult rats. Rats were undernourished by feeding their dams an 8% protein diet from the day of delivery until weaning (21 days of age). The beta-endorphin-like immunoreactivity was measured by radioimmunoassay. In adult rats, undernutrition decreased the basal level hypothalamic beta-endorphin. Avoidance training decreased the content of beta-endorphin in the hypothalamus of well-nourished adults, but had no effect on the levels of previously undernourished rats. In 21-day-old rats, neither undernutrition nor avoidance training altered the levels of beta-endorphin. These results suggest that the hypothalamic beta-endorphinergic system of weaning rats is not yet functional in relation to the parameters analyzed. Probably, other developmental factors are necessary for the emergence of the effects of undernutrition found in adult rats and for the emergence of the response of this system to training (novelty). PMID- 2976161 TI - The isolation of multiple forms of beta-endorphin from the intermediate pituitary of the Australian lungfish, Neoceratodus forsteri. AB - The pituitary of the Australian lungfish, Neoceratodus forsteri, was screened immunohistochemically with heterologous antisera specific for either the C terminal of mammalian beta-endorphin or the acetylated N-terminal of beta endorphin. Immunopositive cells were only detected with the N-terminal specific antiserum; these cells were restricted to the intermediate pituitary. Acid extracts of the intermediate pituitary were fractionated by Sephadex gel filtration chromatography, CM cation exchange chromatography and reverse phase HPLC. Fractions were analyzed by radioimmunoassay (RIA) with a N-acetyl specific beta-endorphin RIA and by radioreceptor assay for the presence of opiate active forms of beta-endorphin. Both immunoreactive and opiate active forms of beta endorphin were detected. Of the total beta-endorphin-related material isolated from the intermediate pituitary, approximately 97% was detected with the N terminal specific RIA and approximately 3% was detected by the radioreceptor assay. The N-acetylated immunoreactive beta-endorphin could be separated into two forms. The major form had an apparent molecular weight of 3.2 Kda. This material had a net charge at pH 2.5 of +5. The minor form of immunoreactive beta-endorphin had an apparent molecular weight of 1.4 Kda and a net charge at pH 2.5 of +1. Neither immunoreactive form exhibited receptor binding activity in the radioreceptor assay. A single peak of opiate active beta-endorphin was detected. This material had an apparent molecular weight of 3.5 Kda and a net charge at pH 2.5 of +7. PMID- 2976162 TI - Atrial natriuretic factor (ANF) gene expression in the Brattleboro rat. AB - Atrial natriuretic factor (ANF) is a 28-amino acid peptide hormone of cardiac origin. It has natriuretic, diuretic and vasorelaxant properties and inhibits several cardiovascular modulators. Because of the possible effects of arginine vasopressin (AVP) on ANF secretion, we have investigated ANF gene expression in Brattleboro rats which are genetically deficient in AVP. Our results indicate that cardiac ANF mRNA and ANF content are higher in Brattleboro rats compared to Long-Evans controls, whereas the plasma levels are similar in both groups. Typical secretory granules containing immunoreactive ANF are present in ventricular cardiocytes of Brattleboro but not of Long-Evans rats. These data suggest that ANF release may be uncoupled from its synthesis in the absence of AVP. PMID- 2976163 TI - [Results of the treatment of patients with advanced breast with large doses of medroxyprogesterone acetate]. PMID- 2976164 TI - Non-opiate effects of neuropeptides derived from beta-endorphin. AB - Brain enzymes convert the opioid peptide beta-endorphin (beta E-(1-31)) to alpha- and gamma-endorphin and to several non-opioid fragments by further cleavage of the tyrosine residue or acetylation. Several of these peptides selectively affect brain functions. alpha-Endorphin (beta E-(1-16)) and relate non-opioid fragments (beta E-(2-16), beta E-(2-9) a.o.) like amphetamine, delay extinction of pole jumping avoidance behavior and facilitate passive avoidance behavior. In addition these peptides enhance the stereotyped sniffing response induced by the injection of apomorphine into the nucleus caudatus. The fragment beta E-(10-16) inhibits, like serotonin and antidepressants, the behavioral effects of melatonin injected into the nucleus accumbens. gamma-Endorphin (beta E-(1-17)) has inherent opioid and neuroleptic-like properties, e.g. demonstrated by a naloxone reversible inhibition of hypermotility induced by apomorphine following injection into the nucleus accumbens. Also the non-opioid gamma-type endorphins (e.g. DT gamma E (beta E-(2-17) and DE gamma E (beta E-(6-17)) mimic certain effects of neuroleptics. These peptides facilitate extinction of pole-jumping avoidance behavior, attenuate passive avoidance behavior and antagonize the hypomotility and stereotyped sniffing induced by apomorphine injected into the nucleus accumbens and pyriform cortex respectively. These and other behavioral studies, including grasping responses, brain stimulation reward, food and other positively rewarded behavior, indicate that the action of alpha-type endorphins is in some aspects comparable to that of psychostimulants, while the effects of gamma-type endorphins are comparable to those of classical as well as atypical neuroleptics. Indeed, gamma-type endorphins have antipsychotic effects in a category of schizophrenic patients. PMID- 2976165 TI - [A fantasy game as group psychotherapy for handicapped children]. PMID- 2976166 TI - Medroxyprogesterone acetate and tamoxifen do not decrease aggressive behavior in CF-1 male mice. AB - Intact CF-1 male mice were given daily injections of either medroxyprogesterone acetate (MPA, an antiandrogen), tamoxifen (TAM, an antiestrogen), or the two drugs in combination and tested for aggressive behavior toward bulbectomized stimulus males. None of the treatments decreased fighting behavior over a 20-day test period and the presence of TAM led to increased aggression even in the presence of MPA. Testis weight was reduced by MPA while both compounds decreased seminal vesicle weight. The mechanisms involved in the observed effects are considered as well as the implications of the results for the clinical use of these compounds as modulators of testosterone-facilitated behaviors. PMID- 2976167 TI - Effects of prenatal 5-methoxytryptamine and parachlorophenylalanine on serotonergic uptake and behavior in the neonatal rat. AB - Parachlorophenylalanine (pCPA) or 5-methoxytryptamine (5MT) was administered to pregnant Sprague Dawley rats from day 8 (D8) of gestation till D17 and from D12 until birth respectively. Birth weights of both drug groups of neonates were approximately 20% less than the saline-injected controls. 5MT neonates showed a significant reduction of high affinity 3H-5HT uptake in the brainstem at all three time points: D1, D15, D30, and a slight reduction in the forebrain reaching significance only on D30. The pCPA animals showed a significant reduction in the brainstem and forebrain on D1 and D30, but only a small nonsignificant reduction in both areas on D15. Behaviors measured on day 15 revealed that in general activity, spontaneous alternation, and passive avoidance both drug groups of neonates showed deficits: less activity, less alternation, and less avoidance. PMID- 2976168 TI - Opioid influence on some aspects of stereotyped behavior induced by repeated amphetamine treatment. AB - Rats were administered repeated IP injections of dl-amphetamine (AMPH) according to a chronic escalating dose schedule (three doses per 24 hr, for four days, two days or one day). Animals treated for four days exhibited a diminished oral stereotypy in response to a challenge of 12 mg/kg AMPH or 2 mg/kg SC apomorphine (APO), 72 hr after withdrawal. Pretreatment with 2 mg/kg IP naloxone (NAL) during the period of chronic AMPH administration prevented the reduction in oral stereotypy induced by AMPH or APO. No differences were detected among the mean of stereotypy scores from the different treatments in response to a challenge dose of 6 mg/kg AMPH. Neurochemical data showed that NAL pretreatment reversed the depletion of striatal dopamine content induced by chronic AMPH. When repeated injections of AMPH were given only one day, the diminished stereotypy response to AMPH or APO was not observed. Animals treated simultaneously with 1 mg/kg IP morphine or 5 micrograms/kg IP beta-endorphin and repeated AMPH injections for one day, showed a reduced stereotyped response to AMPH or APO. These results suggest that opioid peptides are involved in the mechanisms underlying the decrease in oral behaviors following AMPH treatment. PMID- 2976169 TI - Effects of ethanol withdrawal on beta-endorphin levels in rat brain and pituitary. AB - Rats which received a liquid diet containing 6.5% (w/v) ethanol for three weeks became tolerant to the hypothermic effect of an acute dose of ethanol. Withdrawal of this diet was followed by loss of the tolerance within 3 days, and by an accompanying pattern of changes in levels of immunoreactive-ir-beta-endorphin in several brain regions. An initial decrease in levels on days 1 and 3 of withdrawal was followed by recovery to control levels on days 8 and 15. This pattern was found in the arcuate nucleus, amygdala, septum, periventricular thalamus and pre-optic periventricular hypothalamus (POPH), but was statistically significant only in the POPH. A different pattern of change in ir-beta-endorphin levels was found in the pituitary. The anterior lobe showed a significant depletion of ir-beta-endorphin levels before alcohol withdrawal, which recovered by day 8 of withdrawal. This depletion was probably not related to the loss of tolerance to ethanol but was a response to a perturbation of the hypothalamic pituitary axis of hormonal control. PMID- 2976170 TI - Antihypertensive profile of cicletanine, a furopyridine derivative: comparison with captopril, indapamide and prazosin. AB - The effects of cicletanine were compared with those of three other antihypertensive drugs: prazosin, a highly selective alpha 1 antagonist, captopril an angiotensin converting enzyme inhibitor and indapamide a diuretic antihypertensive agent, on young stroke-prone SHR rats with high salt diet; furthermore, vascular reactivity to cicletanine was studied on isolated rat aorta. At an equal dose (30 mg/kg per os) all the drugs prevent the onset of hypertension with the same intensity. The minimal effective dose on blood pressure was 1 mg/kg for both cicletanine and captopril, and 3 mg/kg for indapamide. The action on diuresis and electrolyte excretion occurs at a dose of cicletanine 10 to 30 times higher than that required to produce the anti hypertensive effect. One of the possible mechanisms of the antihypertensive effects of cicletanine could be due to a direct action of the drug on the vascular wall. This vascular impact could be an interaction with the alpha adrenoceptor system (apparent pA2 cicletanine = 5.12) or a decrease in the vascular spasmogenic response whatever agonist was studied. PMID- 2976171 TI - Pathophysiological role of atrial natriuretic peptide in man. PMID- 2976172 TI - [How can bivalves continue to close the shells?--regulation and a molecular mechanism on the economical catch contraction]. PMID- 2976173 TI - Auditory disorders in Down's syndrome. AB - This paper reports a study of hearing and middle-ear function in 100 Down's Syndrome individuals, and the findings are compared with those of previous studies. The high incidence of middle-ear dysfunction and the associated conductive hearing loss, and the progressive sensorineural loss frequently found in Down's Syndrome are discussed. The role of surgical treatment and the use of hearing aids in the management of deafness in Down's Syndrome are reviewed. PMID- 2976174 TI - Liver-specific growth factors. AB - Experimental evidence of the existence of liver-specific growth factors has been collected for more than two decades. Blood-borne growth-promoting activity of hepatocytes may be separated into plasma and platelet-derived factors. Several groups have observed the stimulation of hepatocyte growth in vitro by some platelet-associated activity, which was recently isolated from rat platelets as a 27-kDa protein called platelet growth factor (PGF). There is evidence of at least two different growth factors for hepatocytes derived from platelet-poor rat plasma, 'hepatopoietin' A and B. The partial purification of several other factors has been reported. One of these factors was prepared from the plasma of patients with fulminant hepatic failure. In addition to these 'humoral' factors, cytosolic growth-promoting activity has been partially purified by several groups. While the humoral factors described so far are only active on normal hepatocytes, the cytosolic 'hepatic stimulator substance' (HSS) also promotes the proliferation of differentiated hepatoma cells. In addition, it appears to depend on the permissive action of epidermal growth factor (EGF). None of the liver specific growth factors except PGF has been purified to homogeneity. Thus, their significance for the control of the proliferation of normal and transformed hepatocytes is still an unsettled issue. PMID- 2976175 TI - Pathophysiology and clinical consequences of metabolic alkalosis in hemodialyzed patients. PMID- 2976176 TI - [Radiologic exploration of impotence]. PMID- 2976177 TI - Occluding structures in circumflex veins of the penis. PMID- 2976178 TI - Prolonged penile erection following papaverine--experiences with prophylaxis and therapy. PMID- 2976179 TI - Sexual arousability and sexual orientation of men with erectile dysfunctions. PMID- 2976180 TI - [NMR and cavernosography in the staging of Peyronie's disease]. PMID- 2976181 TI - Caverno-venous leakage: retrograde opacification and resection of the deep dorsal vein completed by ligation of the crural edges of the corpora. PMID- 2976182 TI - [Spontaneous rupture of the kidney. Wunderlich syndrome]. PMID- 2976183 TI - [Testicular metastasis of prostatic carcinoma: presentation of a case and a review of the literature]. PMID- 2976184 TI - [A new type of suprapubic cannula for continuous drainage during transurethral electro-resection of the prostate]. PMID- 2976185 TI - Fluid intake for renal stone prophylaxis. PMID- 2976186 TI - Infected renal stones and defects of ureteral peristalsis. PMID- 2976187 TI - Infection induced renal stones: etiopathological remarks. PMID- 2976188 TI - Staging of renal cancer: CT versus angiography. PMID- 2976189 TI - [Prophylaxis of idiopathic kidney calculi with oligo-mineral water therapy]. PMID- 2976190 TI - [Diagnostic accuracy, sensitivity and specificity of computerized tomography in the staging of renal carcinoma]. PMID- 2976191 TI - [Clinical aspects of dermatologic lesions of the genital area in children]. PMID- 2976192 TI - [Arguments against the recognition of arthroses of the knee joint following occupational stress as an occupational disease]. PMID- 2976193 TI - [Seven cases of emphysema during dental treatment]. PMID- 2976194 TI - Plural medicine in Sri Lanka: do Ayurvedic and Western medical practices differ? AB - In Sri Lanka, as in India, two formally structured systems of medicine exist side by side. While Western-style biomedicine is believed to be useful, Ayurvedic medicine is well established and commonly used. Underlying one explanation for the persistence of such plural medical systems is a functional theory, suggesting that each system is used for different treatments, diseases, or for the ideological, linguistic or social characteristics of the physician. In part, Ayurvedic and Western medicine may persist because their practitioners provide distinctly different services. We tested part of this functional explanation by sending trained 'pseudo-patients' to 764 Ayurvedic and allopathic physicians across Sri Lanka. 'Patients' reported symptoms of common cold, diarrhea or back pain, and recorded after leaving the clinic many aspects of history-taking, diagnostic procedures and physician-patient interaction. Medicines prescribed were later analyzed by a laboratory. We found, basically, no significant differences between the medical practices of sampled Ayurvedic and Western-style physicians, with one exception. While both types spend 3-4 min asking four questions and doing two or three physical examination procedures, and while both prescribe, overwhelmingly, only Western medicines, the allopathic physicians give drugs, that, from the point of view of Western medicine, either 'help' or 'harm' and Ayurvedic physicians prescribe 'neutral' medicines. While we have not directly tested the entire functional explanation we suggest that a structural explanation of the persistence of two systems of medicine may be more valid. Ayurvedic and Western medicine continue in Sri Lanka because they, as institutions, are linked to the social, economic and political structure of the society. Thus, survival is based, not on what a physician does in his practice but upon the power of his medical profession to control medical territory. PMID- 2976195 TI - [A system for analyzing the organization of physiotherapeutic services for the population of large cities]. PMID- 2976196 TI - [Mass screening of schoolchildren with various non-infectious diseases of the digestive organs in the Kirghiz SSR]. PMID- 2976197 TI - [Prevalence of smoking among guests at health resorts and ways of its reduction]. PMID- 2976198 TI - [Creating a network of ambulatory care facilities and their distribution in rural communities of the Azerbaijan SSR]. PMID- 2976199 TI - [A speeding-up strategy and current problems of economic education of public health personnel]. PMID- 2976200 TI - [Prospects of the development of geriatric services]. PMID- 2976201 TI - [Increasing economic effectiveness of the use of medical equipment]. PMID- 2976202 TI - [70th anniversary of the founding of the People's Commissariat of Public Health of the RSFSR]. PMID- 2976203 TI - [Improving the nephrological services]. PMID- 2976204 TI - [Evolution of the views on the organization of the treatment of esophageal cancer]. PMID- 2976205 TI - [Atopic diathesis and the incidence of acute respiratory infections in children of preschool age]. PMID- 2976206 TI - Tissue fluid penetration and anti-treponemal activity of trospectomycin (a new spectinomycin analog) in the rabbit syphilis model. AB - The antitreponemal activity of trospectomycin (a novel 6'-propyl analog of spectinomycin) was correlated with concentrations in serum and tissue chamber fluid in a cutaneously-infected rabbit model of syphilis. After single-dose intramuscular (im) injections of trospectomycin, spectinomycin hydrochloride, and aqueous procaine penicillin G(APPG), concentrations of the drugs in paired specimens of serum and tissue fluid were determined and correlated with response of Treponema pallidum-infected lesions. Lesions responded most rapidly in APPG treated animals. Rapid response correlated with more prolonged levels of APPG in both serum and tissue fluid. Trospectomycin, when given in single doses exceeding 40 mg/kg, surpassed penicillin in peak serum levels (greater than 60 micrograms/ml at 1 hr) but not in duration of activity. Higher and prolonged concentrations of trospectomycin in tissue fluid correlated with more effective clearance of treponemes from cutaneous lesions. Animals treated with less than 40 mg of trospectomycin/kg or with 20-80 mg/kg of the parent compound (spectinomycin hydrochloride) had cutaneous lesions that were persistently darkfield-positive, as confirmed by three or more consecutive smears. PMID- 2976207 TI - Cutaneous fixed drug eruptions from systemic use of drugs. PMID- 2976209 TI - Workers' compensation and psychiatric disability. AB - Psychiatric claims under workers' compensation law have increased substantially in recent years. This chapter explores the history and varying jurisdictional standards of workers' compensation law followed by a discussion of issues pertinent to performing a psychiatric evaluation of a claimant for workers' compensation. PMID- 2976208 TI - Antimicrobial susceptibility of Neisseria gonorrhoeae isolates from a military population in San Diego. AB - Neisseria gonorrhoeae isolates were studied to determine their patterns of antimicrobial susceptibility and possible chemotherapeutic implications. Of 370 consecutive isolates, 32 (8.7%) were penicillinase-producing N. gonorrhoeae (PPNG). The remaining 338 were subjected to disk-diffusion tests, and those apparently resistant to penicillin, tetracycline, or spectinomycin were tested by an agar-dilution method. The dilution test showed that 5.4% (20/370) were penicillin-resistant, non-PPNG strains, of which 100%, 90%, and 45% were also resistant to tetracycline, cefoxitin, and erythromycin, respectively. No resistance to spectinomycin or ceftriaxone was demonstrated, although there was an association between minimum inhibitor concentrations (MICs) of penicillin of greater than or equal to 1.0 microgram/ml and increased MICs of ceftriaxone. The overall incidence of penicillin resistant isolates, including PPNG, was 14.1% (52/370). Of the 20 penicillin-resistant, non-PPNG strains, all were also resistant to tetracycline, and another 21 exhibited tetracycline resistance but were sensitive to penicillin. The in-vitro data suggested that: (1) neither penicillin, tetracycline, nor cefoxitin were acceptable drugs for routine treatment of gonorrhea in our population during the study period; (2) spectinomycin and ceftriaxone continue to demonstrate adequate in-vitro activity against N. gonorrhoeae despite increasing in-vitro resistance to penicillin; and (3) non-plasmid-mediated resistance to penicillin may predict future resistance to ceftriaxone. PMID- 2976210 TI - Cumulative injury in workers' compensation. AB - Over the past few decades a new type of claim against workers' compensation has been steadily evolving, namely cumulative injury claims. The author examines the cost of cumulative injury claims, discusses the issue of causation, and reviews cumulative injury claims with respect to current law. PMID- 2976211 TI - [Advantages and shortcomings of platelet antiaggregants in the treatment of myocardial infarction]. AB - The authors compared the action of small doses of aspirin (100-200 mg/day) and ticlopidine (500 mg/day) in multimodality therapy of acute myocardial infarction. Rapid normalization of aggregation time and disaggregation percentage, a sharp decrease in spontaneous aggregation before adding ADP were noted on the first 5 days of ticlopidine therapy. Platelet aggregation indices returned to normal 3-4 days earlier in the use of ticlopidine versus aspirin. However ticlopidine administration over 7 days was characterized by more frequent episodes of G. I. tract hemorrhages of short duration. There were also some data on the toxic effect of the drug on hepatocytes. More allergic reactions were observed in comparison with the group of patients on aspirin therapy. In order to achieve an optimal antiaggregation effect and to reduce the number of ticlopidine-related side effects short-term courses of ticlopidine with subsequent administration of aspirin at small doses were recommended. PMID- 2976212 TI - [Principles of the use of anti-influenza drugs]. AB - Long-term experience in the clinical and epidemiological testing of antiinfluenzal drugs made it possible to derive major principles of their practical application. A study of the therapeutic and prophylactic efficacy of antiinfluenzal drugs has demonstrated convincingly that success of testing depends on the onset of treatment and epidemiological substantiation of drug administration. The observance of these conditions provides a social, medical and economic effect. Practical implementation of the results obtained required the identification of risk groups of developing influenza and other ARVI, the expansion of a network of rapid diagnostic laboratories, early emergency prevention with maximum coverage of high risk groups, and early administration of antiinfluenzal drugs distributed free of charge. PMID- 2976213 TI - After the tragedy: living with the consequences. PMID- 2976214 TI - [Management of clinical or biological abnormalities during therapeutic trials]. PMID- 2976216 TI - [The status and activities of Soviet forensic medical expertise in the 20th century]. PMID- 2976215 TI - [Mechanisms of cutaneous accidents related to non-steroidal anti-inflammatory agents]. PMID- 2976217 TI - [Radiotherapy of laryngeal cancer at oncological institutions of Volgograd and the Volgograd district]. PMID- 2976218 TI - [In memory of Prof. Andrei Gavrilovich Likhachev, Corresponding Member of the Academy of Medical Sciences of the USSR and honored scientist of the RSFSR]. PMID- 2976219 TI - [With initiators of competition]. PMID- 2976220 TI - Managing low back pain--a comparison of the beliefs and behaviors of family physicians and chiropractors. AB - Random samples of 605 family physicians and 299 chiropractors in Washington were surveyed to determine their beliefs about back pain and how they would respond to three hypothetic patients with back pain. With 79% of the family physicians and 70% of the chiropractors responding, family physicians and chiropractors differed greatly not only in their technical approaches to back pain--such as drug therapy versus spinal manipulation--but also in their underlying beliefs and attitudes. Family physicians think that most back pain is caused by muscle strain, that lumbosacral radiographs are rarely useful, that appropriate therapy does not depend on a precise diagnosis, and that back pain will usually resolve within a few weeks without professional help. Family physicians were more likely than chiropractors to feel frustrated by patients with back pain, less likely to think they can help patients prevent future episodes of back pain, and less confident that their patients are satisfied with their care. Studies are needed to determine whether the different perspectives of family physicians and chiropractors are associated with differences in the costs and outcomes of care. PMID- 2976221 TI - [New results from the literature on partnership development of handicapped children and adolescents]. PMID- 2976222 TI - Oestrus phases of the Syrian hamster. PMID- 2976223 TI - [Comparison of 2 simple surgical methods: posterior colpoceliotomy versus laparoscopy--competition or synergism?]. AB - Celiocolpotomy and laparoscopy often have the same indications, such as oviduct sterilization or diagnostical inspection of the inner genital organs. In order to evaluate their significance in comparison, we examined the course of operation as well as postoperative development in 773 cases in our department in which either a celiocolpotomy (CCP n = 441) or laparoscopy (n = 332) had been performed. Both surgical methods were compared regarding indication, intra- and postoperative complications as well as surgical advantages an disadvantages. Concerning severe intraoperative complications and "failures" both methods appeared equivalent. The rate of--mostly bland--postoperative disturbances (urinary tract infection, rises of temperature) was higher in the CCP group. Regarding the results, it seems recommendable not to view both methods in competition, but to apply them synergistically: factors such as parity, obesity, prior operations, necessity to inspect the entire abdominal cavity as well as the probability and possibility to perform successfully under circumvention of laparotomy should be carefully valued in deciding which surgical method to chose. PMID- 2976224 TI - The effects of desogestrel and ethinylestradiol combination in normal and hyperandrogenic young girls: speculations on contraception in adolescence. AB - Hormone profile and ovarian morphology were studied in two groups of adolescents (group 1:19 girls with slight signs of hyperandrogenism; group 2: 14 normal adolescents) in basal conditions and during a contraceptive combination of 30 micrograms ethinyl estradiol (EE) and 150 micrograms desogestrel (D). Treatment was associated with a low incidence of side effects in both groups. In group 1, acne generally improved within 12 months while hirsutism was only reduced in some subjects (58%) after 12 months of therapy (basal hair score 8.50 +/- 1.60 vs 5.81 +/- 1.53 p less than 0.001). Significant falls in plasma levels of LH, total and free testosterone and an increase in sex-hormone-binding globulin levels were observed during treatment especially in group 1. High percentage of multifollicular ovaries (75%) characterized hyperandrogenic subjects. Ovarian volume and number of follicles, higher in group 1 than 2 in basal conditions, showed a significant reduction in both groups and normal ovarian morphology was restored in hyperandrogenic subjects. Considering the high incidence of hyperandrogenemia in adolescence and its implications, our data suggest that the EE.D combination suits adolescent biological condition and is one of the suitable contraceptive methods in adolescents which also has therapeutic effects. PMID- 2976225 TI - Transvaginal ultrasound or laparoscopy for oocyte retrieval (experiences in the Ljubljana IVF program). AB - At present, the less invasive ultrasound-directed techniques are the methods of choice for oocyte retrieval in most in vitro fertilization and embryo transfer (FIVET) centers. Among the ultrasound-direct techniques, the transvaginal follicle aspiration guided by transvaginal ultrasound for oocyte recovery is gaining popularity in many FIVET centers. This study compare cycles outcome following transvaginal ultrasound oocyte retrieval (105 cycles) to laparoscopic oocyte retrieval (218 cycles); no statistically significant difference could be demonstrated between the groups in all parameters evaluated but better clinical results have been obtained in transvaginal ultrasound group. The Authors conclude that transvaginal oocyte recovery represents an improvement and a simplification of the FIVET procedure. PMID- 2976226 TI - Gamete intrafallopian transfer: different routes of transfer. AB - The Authors report their own experience and results using different approaches to oocyte pick-up and gamete transfer for gamete intrafallopian transfer (GIFT). The overall pregnancy rate of GIFT is 35.86% (66 pregnancies on 184 cases of GIFT). The Protocol I (Laparoscopic oocyte retrieval and gamete transfer) is presently the more used and gives better clinical results (pregnancy rate of 37.7%); in the Author's opinion, the protocols III (echographic pick-up + IVF + laparoscopic delayed zygote intrafallopian transfer (ZIFT), and VII (Laparotomic pick-up and transfer) are interesting complementary techniques for GIFT. PMID- 2976227 TI - Skeletal muscle pathology in ovine congenital progressive muscular dystrophy. 1. Histopathology and histochemistry. AB - The histopathological lesions of ovine congenital progressive muscular dystrophy (CPMD) were characterized by myofiber hypertrophy, focal myofibrillar degeneration, formation of peripheral and central sarcoplasmic masses devoid of myofibrils and internal nuclei often in chains. Progressive loss of myofibrils was associated with atrophy of the fiber and eventual collapse of the sarcolemma. The process was polyphasic, consequently in mature lesions there was great variation in fiber diameter. Split fibers were common but ring fibers occurred rarely. Myofiber loss was associated with fatty or fibrous tissue replacement. Only type I (red, slow twitch, oxidative) fibers were affected and there was no histological evidence of effective regeneration. Ovine CPMD has many histopathological features in common with dystrophia myotonica in humans. PMID- 2976228 TI - Early postnatal deprivation of active sleep with desipramine or zimeldine impairs later behavioural reactivity to auditory stimuli in rats. AB - To examine the functional significance of early postnatal active sleep for the development of behavioural reactivity to auditory stimuli, rat pups were daily injected i.p. from the 7th to the 18th postnatal days with 5 mg kg-1 (6.6 mmol l 1) desipramine or 25 mg kg-1 (12.2 mmol l-1) zimeldine. Sleep-wake behaviour was recorded with a static-charge-sensitive bed (SCSB) method. Both desipramine and zimeldine suppressed the percentage of active sleep relative to the total recording time throughout the treatment period. In addition, these drugs increased the percentage of quiet state and waking. At the age of 38 days the zimeldine-treated rats showed more motor activity in the open field than the controls. At the age of 39 and 78 days all rat groups behaved similarly in the open field. Startle measures and motor activation, provoked by auditory stimulation, were determined by the SCSB method when the rats were 4 months of age. Auditory stimuli, consisting of a series of ten clicks, induced a greater number of startles as well as strong movement responses in the control rats than in the desipramine- or zimeldine-treated rats. The number of small movement responses did not differ between the rat groups. These findings indicate that early postnatal active sleep and the monoaminergic systems regulating it may be important for the normal development of neuronal circuitry associated with later reactivity to auditory stimuli. PMID- 2976229 TI - Key enzymes of myocardial energy metabolism in patients with valvular heart disease: relation to left ventricular function. AB - Endomyocardial biopsies were taken from the apex of the left ventricle in 15 patients operated on for aortic valve disease or ischaemic heart disease and from papillary muscles in six patients operated on for mitral valve disease. Activities of cardiac phosphofructokinase (PFK), total lactate dehydrogenase (LD), its isoenzyme LD1, aspartate aminotransferase (ASAT), total creatine kinase (CK), its isoenzyme MB, citrate synthase (CS) and myoglobin content (MYO) were related to the angiographically determined left ventricular function. Activities of total LD, PFK and PFK/CS ratio were lower in patients with decreased, than in those with normal, left ventricular function. Myoglobin content and activities of CS and ASAT were not related to left ventricular function. It is suggested that depressed left ventricular contractility is associated with a decreased glycolytic capacity while the oxidative capacity is mainly unaltered. PMID- 2976230 TI - Increase in myoglobin content and decrease in oxidative enzyme activities by leg muscle immobilization in man. AB - Biopsies from m. quadriceps femoris from the operated leg of nine patients were taken before, and 6 weeks after, knee surgery. During the whole postoperative period the operated leg was immobilized with the knee in 40-50 degrees of flexion. Myoglobin (MYO) and the enzymes citrate synthase (CS), creatine kinase (CK) and its isozymes MB (CK-MB) and mitochondrial CK (CK-MIT), aspartate aminotransferase (ASAT), phosphofructokinase (PFK) and lactate dehydrogenase (LD) were determined on the biopsies. Citrate synthase, ASAT, CK, CK-MB, CK-MIT and LD activities were decreased (12-30%) after the postoperative leg immobilization period. Phosphofructokinase did not change, while MYO content was increased (16%). In conclusion, a different control of the synthesis of oxidative enzymes and MYO is suggested, as the induced changes following immobilization were in opposite directions. The function of the increased MYO content may be to facilitate the oxygen extraction. PMID- 2976231 TI - Renal effects of atriopeptin II and dopamine receptor blockade in acutely volume expanded rats. AB - Studies were made of the effects of continuous intravenous infusion of a synthetic atrial natriuretic factor (ANF) or, pre-treatment with the dopamine receptor antagonist haloperidol, on the renal response in anaesthetized rats subjected to volume expansion with an isotonic solution at 2% kg-1 body weight (wt) h-1. A time-control group receiving vehicle alone was studied in parallel. Measurements were compared 75 and 145 min after initiation of the volume expansion. Seventy minutes of Atriopeptin II infusion at 10 micrograms h-1 kg-1 body wt did not significantly alter the glomerular filtration rate [control value 1.29 +/- 0.10 ml min-1 g-1 kidney wt (n = 7, mean +/- 1 SEM), experimental value 1.20 +/- 0.12], but increased sodium excretion by 49% (from 2.87 +/- 0.56 to 4.27 +/- 0.45 mumol min-1). The arterial blood pressure was reduced by 9%. In previous investigations we found that in the same dosage Atriopeptin II increased sodium excretion 10-fold in euvolaemic animals. In the time-control group (n = 7) the response was similar to that in the atrial natriuretic factor-treated animals with the exception that the blood pressure was unaltered. Thus, glomerular filtration rate showed no statistically significant change (1.28 +/- 0.06 vs. 1.27 +/- 0.09 ml min-1 g-1 kidney wt) while the sodium excretion increased by 96% (from 2.29 +/- 0.22 to 4.50 +/- 0.49 mumol min-1). In animals pretreated with haloperidol (n = 5), the natriuretic response to the volume expansion was attenuated and was about ten times below that in the time-control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976232 TI - Electric muscle stimulation in the hind leg of the spontaneously hypertensive rat induces a long-lasting fall in blood pressure. AB - The influence of prolonged low-frequency, low-intensity electric stimulation of the gastrocnemius muscle or of the biceps femoris muscle on blood pressure and heart rate was investigated in unanaesthetized, spontaneously hypertensive rats (SHR). In both groups, elevations of blood pressure and heart rate were elicited during the 60 min of muscle stimulation. After cessation of the stimulation, a depressor response developed within 60 min. Thirty to sixty minutes post stimulation the fall in blood pressure was 19 +/- 3 and 17 +/- 4 mmHg, respectively (mean +/- SE) compared with controls. In both groups, the depressor response lasted for over 5 h. In addition, the gastrocnemius-stimulated animals also developed a post-stimulatory bradycardia. In one group of SHR the sciatic nerve was anaesthetized with bupivacaine. The arousal response during stimulation was similar to that in the other groups, but after termination of stimulation blood pressure returned to the control level without any further drop. To investigate further the neurotransmitters involved, one group of gastrocnemius stimulated SHR was given naloxone by infusion during the stimulation. A modest post-stimulatory blood pressure fall also occurred in this group, but it lasted only 90 min. Another SHR group was pre-treated with parachlorophenylalanine, a serotonin synthesis blocker, which completely abolished the post-stimulatory depressor response. These results indicate that prolonged muscle stimulation gives rise to a post-stimulatory long-lasting drop in blood pressure and that this response is mediated by somatic nerve afferents. Involvement of the endorphin and serotonin systems is also suggested. PMID- 2976234 TI - CNS-induced natriuresis during dopamine receptor blockade. Further support for the existence of, at least, two separate natriuretic hormonal systems. AB - Intracerebroventricular (ICV) stimulation with hypertonic sodium chloride solutions has previously shown indications to stimulate the release of a blood borne natriuretic factor. Sodium excretion in this situation increases in the order of 10-20 times. Acute isotonic volume expansion has shown to be a potent stimulus for endogenous release of the atrial natriuretic factor (ANF). It has also been demonstrated that the natriuretic response to both volume expansion and exogenously applied ANF can be attenuated by the dopamine receptor blocker haloperidol. The present study was performed to investigate if the natriuretic response to ICV stimulation also could be attenuated with haloperidol, thus indicating similar effector mechanisms as for ANF. A first group of anaesthetized animals was, therefore, pre-treated with haloperidol (H) and then ICV stimulated (H-ICV). A second group of animals was subjected to acute isotonic volume expansion (VE, 2% b.w.h.-1) to evoke the documented ANF release. In a third group of animals pre-treatment with haloperidol was followed by volume expansion (H VE). In the H-ICV group there was a more than 30-fold increase in sodium excretion, due to an increase both in urine flow rate (more than sixfold) and in the urinary concentration of sodium (more than fourfold). Potassium excretion increased more than eightfold, urine osmolality was unchanged, and blood pressure increased by 7%. In the VE group sodium excretion increased more than 18-fold, consequent to large increases in urine flow rate (more than 23-fold), while the urinary concentration of sodium tended to decrease. Potassium excretion increased more than threefold, urine osmolality decreased by 88%, and blood pressure was unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976233 TI - Transmural distribution of biochemical markers of total protein and collagen synthesis, myocardial contraction speed and capillary density in the rat left ventricle in angiotensin II-induced hypertension. AB - The effect of angiotensin II-induced hypertension on selected biochemical parameters was studied in Sprague-Dawley rats. Angiotensin II infusion at rates of 41.7 micrograms h-1 kg-1 and 12.5 micrograms h-1 kg-1 for 2, 5, 10 and 15 days elevated the systolic blood pressure from 143 +/- 7 mmHg to 215-230 mmHg (P less than 0.001) and 185-195 mmHg (P less than 0.001), respectively. The left ventricular weight/body weight ratio increased 10-14% (P less than 0.05) and 23 32% (P less than 0.001) after 2-15 days in rats treated at the lower and higher infusion rates, respectively. Prolyl 4-hydroxylase (PH) activity, a marker of collagen synthesis, was evenly distributed in the left ventricle. PH activity increased by about 100% in both subendocardial and subepicardial layers of the left ventricular wall after angiotensin II infusion for 10 days at 41.7 micrograms h-1 kg-1, but remained unaltered at 12.5 micrograms h-1 kg-1. No change was observed in hydroxyproline concentration. Myosin isoenzymes (V1-V3), which reflect myocardial contractility, were unevenly distributed in the left ventricular wall: the proportion of the fast-turnover isoenzyme (V1) was smaller in the subendocardial layer than in the subepicardial layer. The proportion of V1 decreased after treatment in both layers. Alkaline phosphatase activity, a marker of capillary density, was evenly distributed transmurally in the left ventricular wall. Angiotensin II caused a slight decrease in this activity in both myocardial layers. The results suggest that the elevation of blood pressure leads to transmurally evenly distributed changes in biochemical parameters reflecting collagen synthesis, capillary density and contractile properties of the myocardium. PMID- 2976235 TI - Haemodynamics and plasma ANP (atrial natriuretic peptide) after acute blood volume expansion in normotensive and spontaneously hypertensive rats. AB - Atrial natriuretic peptide (ANP) was measured in plasma during acute volume load in conscious, spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats. During basal conditions immunoreactive ANP were similar in the SHR (630 +/- 56 pmoles l-1) and the WKY (657 +/- 114 pmoles l-1) groups. An acute 10% and 20% whole blood volume expansion resulted in a linear increase in immunoreactive plasma ANP in the WKY. In the SHR the increase in plasma ANP was attenuated during the 20% volume load. During the 10% and 20% volume load central venous pressure (CVP), central blood volume (CBV) and cardiac output increased relatively more in the SHR compared with the WKY group. In contrast, the increase in peripheral blood volume (PBV) and decrease in heart rate (HR) was attenuated in the SH rats. In the SHR group there was a shift of the ANP vs. CVP and ANP vs. CBV curves to the right compared with the WKY. We conclude that acute volume loading is a potent stimulus for ANP release in WKY as well as SHR. However, in the SHR, ANP release was blunted in spite of the increased centralization of the volume load in this rat strain. Thus, the decreased responsiveness of the ANP hormonal system may contribute to the development and maintenance of hypertension in this genetic form of hypertension. PMID- 2976236 TI - Further characterization of the contraction-mediating prostanoid receptors in feline cerebral arteries. Effects of the thromboxane-receptor antagonist AH 23848. AB - The effects of the thromboxane-receptor antagonist AH 23848 were investigated on isolated feline basilar arteries (BA). AH 23848 (10(-6) mol l-1) had no effect on contractions induced by 5-hydroxytryptamine or potassium, whereas the drug (10( 8)-10(-6) mol l-1) induced a parallel shift to the right in contractions induced by the thromboxane A2 mimic U46619. There was no depression of the maximum contraction, indicating competitive antagonism. The Schild plot revealed a slope index of unity with a pA2 value of 8.46. In contrast, 10(-6) mol l-1 AH 23848 depressed the maximum PGF2 alpha-induced contraction significantly from 100% to 13%. U46619 was able to induce a contraction amounting to 98% if the drug was added on top of the PGF2 alpha-induced contraction in the presence of 10(-6) mol l-1 AH 23848. The results provide strong support for previous suggestions that prostanoid-induced contractions in the feline BA are mediated by two receptor subtypes, one of which can be classified as a thromboxane-sensitive (TP) receptor. PMID- 2976238 TI - The release mechanism for atrial natriuretic factor during blood volume expansion and tachycardia in dogs. AB - Atrial natriuretic factor (ANF) is released during blood volume expansion and tachycardia, but only blood volume expansion causes atrial distension, which presumably promotes ANF release. Our study was undertaken to search for a common release mechanism. In five anaesthetized, closed-chest dogs, plasma immunoreactive (IR) ANF was measured at three levels of blood volume, which were obtained by infusing a Ringer's solution. At each level of blood volume, plasma IR-ANF was measured at three pacing frequencies. Plasma IR-ANF increased as mean right atrial pressure (mRAP) was raised from 2 to 10 mmHg by volume expansion, whereas pacing tachycardia (at heart rates (HR) 50 +/- 3 and 98 +/- 1 beats min-1 above control) at each level of blood volume expansion increased plasma IR-ANF and systolic RAP (sRAP) at constant mRAP. Plasma IR-ANF was more strongly correlated to sRAP (r = 0.83) than to mRAP (r = 0.69), but the product sRAP x HR had the highest correlation coefficient (r = 0.86). According to the multiple regression equation: plasma IR-ANF = k1 + k2mRAP + k3sRAP + k4sRAP x HR, the product sRAP x HR had the highest coefficient of determination (r2 = 0.75) and was the only significant determinant. We conclude that atrial tension or stress, developing during each atrial systole, is an important determinant of ANF release. Since atrial diastolic and systolic dimensions do not increase during pacing tachycardia, ANF release is not dependent on atrial distension. PMID- 2976237 TI - Differential haemodynamic effects of atrial natriuretic peptide (ANP) in normotensive and spontaneously hypertensive rats. AB - Central haemodynamic parameters and cardiac performance were measured in conscious spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto (WKY) control rats after a 10-min infusion of rat ANP (103-125), 1 micrograms kg 1 min-1. Mean Arterial blood pressure (MAP) decreased by approximately 10% in both groups of rats. Heart rate (HR) increased slightly in both strains during the infusion. In the normotensive group the fall in MAP was due to a reduction in cardiac output (CO) while in the SHR there was a decrease in CO as well as in total peripheral resistance (TPR). The ANP infusion also reduced central blood volume (CBV) and stroke volume (SV) in both groups of rats. The reduction in CBV and CO was significantly more pronounced in the WKY strain. Left ventricular end diastolic pressure (LVEDP) and cardiac contractility (dP/dt) did not change while central venous pressure (CVP) was slightly decreased in the WKY group as a result of the ANP infusion. We conclude that ANP reduces MAP in normotensive animals by a reduction in CO. In the SHR a reduction in TPR also contributes to the fall in MAP. Atrial natriuretic peptide did not exert any negative inotropic effects, but the reduction of CO was due to an increased venous compliance. PMID- 2976239 TI - Increased behavioural response to intrathecal serotonin after lesion of serotonergic pathways with 5,7-dihydroxytryptamine seems not to be due to depletion of serotonin. AB - The behavioural response to intrathecal i.th. serotonin (5-HT) was examined in mice pretreated with the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) or the 5-HT synthesis inhibitor p-chlorophenylalanine (PCPA) which produce similar extensive depletion of central 5-HT levels. Intrathecal 5-HT (0.4 micrograms) elicited a behavioural response consisting of reciprocal hindlimb scratching and biting or licking of the hindquarters indicative of nociceptive stimulation. The response to i.th. 5-HT was markedly increased 5 days after intracerebroventricular (i.c.v.) injection of 5,7-DHT (80 micrograms base per mouse). On the other hand, 24 h after the last pretreatment injection of PCPA (400 mg kg-1 for 6 consecutive days), the response to i.th. 5-HT was unaltered. These results indicate that i.c.v. 5,7-DHT produces supersensitivity to 5-HT. Since PCPA failed to alter the effect of 5-HT, the supersensitivity seems not to be due to depletion of 5-HT levels after the lesion. PMID- 2976240 TI - Do the fibre-type proportion and the angular velocity influence the mean power frequency of the electromyogram? AB - The dependence of the mean-power frequency and the signal amplitude of the electromyogram (EMG) on the angular velocity and the fiber-type proportion were investigated in nine female volunteers. The subjects were required to perform maximum knee extensions using an isokinetic dynamometer at different angular velocities; 0.57, 1.05, 1.57, 2.09 and 3.14 rad s-1. Electromyographic signals were obtained from the vastus lateralis, vastus medialis and the rectus femoris muscles. The angle and the torque signals were recorded simultaneously with the three EMG signals on a tape-recorder. From the EMG recordings the mean power frequency (MPF) and the signal amplitude were determined. Muscle biopsies were later obtained from the right vastus lateralis and stained for alkaline and acid mATPase for the determination of fibre-type proportions and areas. Neither the signal amplitude nor the MPF of the EMG of the three muscles were dependent on the angular velocity. The MPF of the vastus lateralis correlated significantly (r = -0.93) with the type 1 fibre proportion at 1.57 rad s-1. However, there was no significant correlation between the areas of the fibre types, alone or together, and the MPF. In conclusion the fibre-type proportion was the major factor behind the MPF irrespective of angular velocity. PMID- 2976241 TI - Vasodilator responses to alpha-human-atrial natriuretic peptide in isolated omental and pulmonary arteries from rabbit and man. AB - In isolated vessels from the pulmonary and mesenteric/omental arterial circulations of rabbit and man, the effects of synthetic alpha-human atrial natriuretic peptide (alpha-hANP) were investigated. The vessels had an outer diameter of 0.6-1.2 mm and were contracted by noradrenaline (NA) and prostaglandin (PG) F2 alpha (mesenteric/omental arteries) or by 5 hydroxytryptamine (5-HT) and PGF2 alpha (pulmonary arteries). It was found that in rabbits, mesenteric vessels contracted by NA and PGF2 alpha were not significantly relaxed by alpha-hANP. Rabbit pulmonary vessels contracted by PGF2 alpha or 5-HT were concentration-dependently relaxed by alpha-hANP (maximum 70%). Human omental vessels contracted by NA and PGF2 alpha showed a moderate (30%) relaxation after addition of the peptide. Human pulmonary arteries contracted by 5-HT were relaxed by alpha-hANP almost to baseline, while PGF2 alpha-contracted vessels showed a maximum relaxation of 65%. It is concluded that alpha-hANP has a relatively selective effect on pulmonary arterial vessels from rabbit as well as man. The results suggest that the peptide may be involved in the regulation of pulmonary vascular tone. PMID- 2976242 TI - Apparent hyperalgesia in the mouse tail-flick test due to increased tail skin temperature after lesioning of serotonergic pathways. AB - The relationship between tail skin temperature and responsiveness to noxious radiant heat in the tail-flick test was investigated in mice. A significant negative correlation between tail skin temperature and tail-flick latency was found when the tail skin temperature was increased by elevating the ambient temperature. After intracerebroventricular injection of the serotonin neurotoxin 5,7-dihydroxytryptamine (5,7-DHT, 80 micrograms) tail skin temperatures were increased and tail-flick latencies reduced. In contrast, administration of the tryptophan hydroxylase inhibitor p-chlorophenylalanine (PCPA, 400 mg kg-1 for 5 consecutive days) lead to a slight lowering of tail temperatures and a tendency towards elevation of tail-flick latencies. The results show that factors which affect tail skin temperature also influence the tail-flick test in mice. The divergent effects of 5,7-DHT and PCPA on tail-flick responsiveness may be due to the different effects of these compounds on the tail skin temperature. The results suggest that the reduced tail-flick latency after partial destruction of serotonergic pathways by 5,7-DHT is due primarily to the increased tail skin temperature. The dependence of tail-flick latency on tail skin temperature limits the usefulness of the tail-flick test unless changes in tail skin temperature are controlled for. PMID- 2976244 TI - Time relation between changes in central venous pressure and the secretion of atrial natriuretic peptide in humans. PMID- 2976243 TI - Atrial natriuretic factor, urinary catechol compounds and electrolyte excretion in rats during normal hydration and isotonic volume expansion. Influence of dopamine receptor blockade. AB - To investigate the influence of acute isotonic volume expansion (VE) on the plasma concentration of atrial natriuretic factor (ANF), the excretion of catechol compounds and electrolytes and the whole kidney glomerular filtration rate (GFR), these variables were measured before and during 60 min of VE (2% of body weight per hour). Atrial natriuretic factor was measured at the end of the experiment. In a control group (n = 7) without volume expansion, plasma ANF was 58 +/- 4 pg ml-1. The excretion of sodium, dopamine (DA), 3,4 dihydroxyphenylacetic acid (DOPAC), noradrenaline (NA) and GFR did not change during the control study. In VE animals (n = 7) plasma ANF was 82 +/- 7 pg ml-1, significantly higher than in the control group. Sodium excretion increased more than 17-fold. The excretion of the DA increased by 38% and that of DOPAC by 30%. Noradrenaline excretion remained unchanged while GFR increased by 20%. In haloperidol-pretreated animals subjected to VE (n = 7), plasma ANF was 81 +/- 8 pg ml-1 during VE, significantly higher than in the control animals. Although the sodium excretion increased more than ninefold in this group during VE, this increase was only 55% of that in the VE group not given haloperidol. The DA and DOPAC excretion was increased by haloperidol, indicating a feedback effect of receptor blockade. DOPAC excretion was not increased further by VE, but the excretion of DA increased by 15% and GFR increased by 19%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976246 TI - Central D1 dopamine receptors. Papers from a symposium. New York, New York, December 5, 1986. PMID- 2976245 TI - Androstenedione concentrations following dexamethasone suppression: correlation with clomiphene responsiveness in women with polycystic ovary syndrome. AB - It is difficult to predict clomiphene responsiveness in women with polycystic ovary syndrome (PCOS) but it has been suggested that women with evidence of excess adrenal androgen are less likely to respond to clomiphene. To investigate this further we performed a short Synacthen test following overnight dexamethasone suppression, using 11 beta-hydroxyandrostenedione (11-OHA) as a specific marker of adrenal androgen secretion in women with anovulatory infertility due to PCOS (n = 19) compared with a normal group (n = 7). Women with PCOS were subsequently divided into 2 groups according to whether or not they ovulated after clomiphene. On day 1 blood was taken at 9.00 hours for measurement of androstenedione (A), 11-OHA and cortisol, and 1 mg dexamethasone was given at 22.00 hours. On day 2 blood was taken at 9.00 hours and at 30 and 60 minutes after intravenous administration of 250 micrograms Synacthen. Before dexamethasone was given, concentrations of A but not of 11-OHA or cortisol were significantly higher in women with PCOS than in controls but there was no difference in A levels between clomiphene responders and non-responders. After 1 mg dexamethasone had been given, concentrations of A, 11-OHA and cortisol were suppressed in all 3 groups and there were no differences between the groups in the post-dexamethasone concentrations of 11-OHA or cortisol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976247 TI - Pharmacological and behavioral effects of D1 dopamine antagonists. AB - This work includes the effects of SCH 23390 and related benzazepines on behavior and attempts to relate these effects to their D1 dopamine antagonist action. Effects on conditioned avoidance responding (CAR) in rats were studied under the same conditions in which in vivo binding of the radioiodinated D1 specific benzazepine 125I-SCH 38840 was measured. It was found that there is very close agreement between the time-course for antagonism of CAR and for in vivo displacement of 125I-SCH 38840 from rat striatum. The effect of SCH 23390 in CAR in monkeys was compared with standard anti-psychotics and although its oral potency was reasonable, its duration was very short (1-2 hours at 5 times its minimal effective dose for statistically significant reduction of avoidance). It is concluded from this and prior work that SCH 23390 and other D1 specific benzazepines inhibit CAR at the same doses that bind to D1 receptors in the CNS and that D1 specific antagonists are behaviorally effective at doses that do not produce D2 receptor effects (e.g. increased plasma prolactin levels, catalepsy). PMID- 2976248 TI - Neurophysiological examination of the role of D-1 dopamine receptors in the regulation of neuronal activity in the basal ganglia. PMID- 2976249 TI - Diminished D2 dopamine receptor function and the emergence of repetitive jaw movements. AB - Oral movements in rats, repetitive jaw movements (RJM), can be induced in a dose dependent manner by a specific D1 agonist, SKF 38393, and decreased by D2 receptor stimulation with a specific D2 agonist, LY 141865. Irreversible D1 receptor inactivation by N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline severely reduced oral responses induced by SKF 38393, whereas such blockade of D2 receptors greatly augmented the D1 mediated behavior. Further, we found that chronic prolonged D2 receptor blockade following administration of fluphenazine decanoate facilitated repetitive jaw movements. PMID- 2976250 TI - D-1 dopamine receptors and arousal. PMID- 2976251 TI - The D-1 dopamine receptor. AB - The D-1 receptor will provide a fruitful ground for many scientists in the coming years. Pure biochemists will attempt to isolate, purify and sequence the molecule itself. Functional biochemists will study the mechanisms whereby the receptor regulates adenylate cyclase activity. Physiologists will attempt to study the consequences of stimulating the receptor in either the brain or in peripheral tissues. Animal behavioralists will attempt to understand how the receptor participates in the generation of animals response to dopaminergic drugs (both agonists and antagonists). Finally, it remains to be determined if any novel therapeutic agents targeted towards the D-1 receptor will become commercially viable compounds. PMID- 2976252 TI - Biochemical properties of D1 and D2 dopamine receptors. AB - The physiological action of dopamine are mediated by two distinct subtypes of receptors, D1 and D2 dopamine receptors. D1-receptors are linked to stimulation of adenylate cyclase whereas D2-receptors inhibit the enzyme and may also couple to other signal transduction systems such as ion channels. In order to characterize these receptors at the biochemical level we have developed specific probes for the identification and purification of these proteins. The ligand binding sites of the two receptors have been identified by photoaffinity labeling and reside on distinct polypeptides. In rat striatum, the D1 receptor binding site can be identified as a peptide of Mr = 72,000. In contrast, the D2 receptors appears to reside on an Mr = 94,000 peptide in most tissues. A larger peptide of Mr = 120,000 identified in the intermediate lobe of pituitary may represent the unproteolyzed form of this receptor. An affinity chromatography purification procedure has been developed for the D2 dopamine receptor. This procedure affords a substantial purification (greater than 1000 fold) of the receptor solubilized from bovine anterior pituitary glands with complete retention of its binding properties. These biochemical tools should eventually lead to the complete characterization of these two receptor subtypes. PMID- 2976253 TI - Biochemical and functional characterization of central dopamine receptors. AB - In order to characterize the D2 dopamine receptors at the molecular level we have developed procedures for purification and characterization of the receptor binding protein. The ligand binding sites of the D2 dopamine receptor have been identified by photoaffinity labeling with 3H-7-azidofluphenazine. The D2 dopamine receptor protein was partially purified by Fast Performance Liquid Chromatography on a Mono Q column and on a wheat-germ agglutinin agarose column. Some behavioral expressions which are probably mediated by D1 dopamine receptors were described. Evidence was obtained that catalepsy is associated with the blockade of D1 dopamine receptors and that D1 and D2 dopamine receptor systems interact either directly or indirectly in mediating this behavior. The administration of a dopamine agonist to monkeys with unilateral ventromedial tegmental lesions of the brainstem produced a biting behavior which, in some aspects, resembles the behavior in Lesch-Nyhan patients. The prevention of the dopamine agonist-induced biting behavior by the D1 dopamine antagonists indicates that D1 dopamine receptors are involved in the control of the expression of this behavior. The biting behavior, like other motor functions, might be controlled by the mesolimbic dopamine systems, and supersensitive mesolimbic D1 dopamine receptors might be associated with the pathology of this behavior. PMID- 2976254 TI - Dopamine D1 and D2 receptor selectivities of agonists and antagonists. AB - The selectivities of various dopamine agonists and antagonists for dopamine D1 and D2 receptors were obtained by comparing their relative dissociation constants for inhibiting the binding of [3H]SCH 23390 at D1 receptors (calf caudate nucleus) and at D2 receptors (pig anterior pituitary tissue). The most selective agonists were SK&F 38393 (for D1) and (+)-PHNO (for D2), while the most selective antagonists were SCH 23390 (for D1) and raclopride or eticlopride (for D2). PMID- 2976255 TI - DARPP-32 as a marker for D-1 dopaminoceptive cells in the rat brain: prenatal development and presence in glial elements (tanycytes) in the basal hypothalamus. AB - The present article reviews some aspects of the localization of a dopamine- and cyclic AMP-regulated phosphoprotein, DARPP-32, which is assumed to be present in D-1 dopaminoceptive neurons. Its prenatal development starts at day 14 of gestation, is to a large extent complete at birth and seems to be independent of ingrowing dopamine-containing afferents. Rearrangements occur in certain areas, and in some systems DARPP-32 appears to be only transiently expressed. The presence of DARPP-32 in glial structures, the tanycytes, in the arcuate nucleus median eminence complex in the mediobasal hypothalamus, has given further support to the hypothesis that dopamine, by controlling the shape of the tanycytes and the extension of their processes, can regulate LHRH release by a 'mechanical mechanism'. This hypothesis is now being examined in some experimental paradigms. PMID- 2976256 TI - Lipocortins. PMID- 2976257 TI - Cellular requirements of IgE-antibody regulation. AB - The low affinity receptor for IgE (Fc epsilon RII), which is identical to CD 23 has been recently implicated in a variety of functions. It appears as an early stage specific marker in the ontogeny of the IgM-bearing B cell. In humans the CD 23 is also exposed on T-cells in patients with elevated IgE, while in rodents Fc epsilon RII seems to be mainly present on T-lymphocytes. Fc epsilon RII can also be detected on macrophages, eosinophils and platelets. Activation of these cells induces an amplification of the inflammatory response. It is currently suggested that IgE-binding molecules, which are related to CD 23, modulate IgE synthesis according to their degree of glycosylation. Evidence has been provided recently that interleukin 4 induces the expression of Fc epsilon RII (CD23). The action of interleukins, IgE binding factors and cytokines on the complex events of IgE induction and synthesis are currently studied. PMID- 2976258 TI - A study of the functions of polymorphonuclear neutrophil in patients with Down's syndrome. AB - Patients with Down's syndrome (DS) are predisposed to infectious diseases, particularly of the respiratory tract. Together with the occurrence of cardiac defects, this is one of the main determinants of their life expectancy. The aim of the present work was to study a series of immunological parameters that would allow us to evaluate the functional behaviour of the neutrophil polymorphonuclears of the patients with trisomy 21 and the possible relationship between this and the susceptibility of this kind of patient to such infections. The results obtained point a significant decrease (p less than 0.001) in the adhesiveness index and in the two indexes relating to the evaluation of random mobilities of cells ("leading front" and "lower face of filter"). Although the neutrophil chemotactic responsiveness of these patients is decreased with any of the attractants employed (casein and activated sera), the differences observed with respect to the control population are more patent upon analyzing the function on the "lower face of filter". Study of phagocytosis of Candida albicans and candidicidal activity of PMN reveals a normal behaviour in the presence of autologous (patient) and heterologous (control) serum. We propose that the alterations found in the PMN cell functions of DS patients could be due to intrinsic cellular defects and that this would help to explain why such patients tend to undergo repetitive infective processes. PMID- 2976259 TI - [T-cell subpopulations and suppressor activity in atopic and infection-induced asthma]. AB - We determined the T-lymphocyte subpopulations by means of monoclonal antibodies (CD 3, CD 4, CD 8) and the so-called suppressor index (increased lymphocyte stimulation after 24 h preincubation) in 20 patients with extrinsic asthma and in 18 patients with intrinsic asthma. In patients with extrinsic asthma we found an increase of ratio CD 4/CD 8. The short-living suppressor activity was significantly decreased in patients with intrinsic asthma. PMID- 2976260 TI - Invited essay on the psychological aspects of genetic counseling. V. Preselection: a family coping strategy in Huntington disease. AB - Preselection refers to the "sick"-role assignment given to an asymptomatic individual in a family in which a genetic or other disorder with multiple uncertainties occurs. The selection process is made, often when the preselected person is still a child, without knowledge of who eventually will be affected. The process requires the collusion of the family members for its initiation and maintenance and serves as a major coping strategy to reduce or bind the stresses and anxieties engendered by uncertainty. Preselection is seen often in families in which Huntington disease occurs and is promoted by the delayed age of onset of the disorder and the uncertain gene status of the person at risk. PMID- 2976261 TI - Development and investigation of the occupational performance history interview. AB - This article describes the development of the Occupational Performance History Interview, an instrument designed to gather an accurate and clinically useful history of an individual's work, play, and self-care performance from psychosocially and/or physically disabled adolescents, adults, and older persons. Following the identification and validation of content and the development of format, the instrument was field-tested and revised. An examination of reliability across time and raters indicated that although part of the instrument ratings met or exceeded levels of acceptable stability, further development and testing is indicated. PMID- 2976262 TI - Hexosaminidase A deficiency manifesting as spinal muscular atrophy of late onset. AB - Proximal lower limb weakness and fasciculations were the only clinical manifestations of hexosaminidase A (Hex A) deficiency in a 39-year-old woman. Hex A activity in serum and leukocytes was 0 to 4% by standard heat inactivation fluorogenic substrate methods, and 5% when Hex A isoenzymes were fractionated by diethylaminoethanol cellulose chromatography. Computed tomography of the brain showed cerebellar atrophy. We suggest that Hex A activity studies be done in patients with typical as well as atypical spinal muscular atrophy. PMID- 2976263 TI - Effects of ribamidine, a 3-carboxamidine derivative of ribavirin, on experimentally induced Phlebovirus infections. AB - Ribamidine (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamidine) was inhibitory in rhesus monkey kidney (LLC-MK2 derivative) cells to Adames and Balliet strains of Punta Toro virus (PTV), a Phlebovirus related to Rift Valley fever and sandfly fever viruses. The 50% effective dose was 8 and 12 micrograms/ml against each respective virus strain; the 50% cytotoxic dose was 320 micrograms/ml, giving selectivity indices of 40 and 27 against each virus strain. The virus ratings were 1.2 and 1.0, respectively. In radiolabel uptake studies, ribamidine had a moderate effect on [3H]leucine uptake at dosages down to 1 microgram/ml, but [3H]thymidine, [32P], and [3H]uridine were inhibited at high (100-1000 micrograms/ml) doses only. Subcutaneous (s.c.) and oral treatments of Adames PTV infected mice were equally highly effective, as evidenced particularly by 100% survivors. Reduced hepatic icterus, serum oxalic acid transaminase, serum glutamic pyruvic acid transaminase, and recoverable virus titers from livers and sera of infected mice were also seen as a result of ribamidine treatment. Twice daily treatment for 5 days could be started as late as 72 h post-virus inoculation (p.v.i.) with significant inhibition of PTV infection seen. Single s.c. treatments administered as late as 48 h p.v.i. were similarly effective. Using the chronic therapy schedule, the maximum tolerated dose was 1000 mg/kg/day and the minimum effective dose was 31.3 to 62.5 mg/kg/day. Using single treatment, a maximum tolerated dose was greater than 1000 mg/kg, and the minimum effective dose was 125 mg/kg. Ribamidine s.c. treatment of mice infected intracerebrally with the Balliet strain of PTV resulted in a moderate infection inhibitory effect, seen especially by reduced virus titers in the brains of the infected, treated mice. PMID- 2976264 TI - Changes in the modulations of kinetics and allosteric properties of muscle phosphofructokinase of young and old rats. AB - In vitro studies on various modulations in kinetics and allosteric properties of muscle phosphofructokinase (EC 2.7.1.11) were undertaken, using purified enzyme from young (25-weeks) and old (100-weeks) albino rats. In comparison to normal K0.5 values for fructose-6-phosphate, increase in this value in response to ATP and citrate inhibition, decrease in K0.5 due to AMP activation and extent of ATP inhibition with increase in pH, were observed to be decreased more markedly with the enzyme of old than with that of young rats. Extent of citrate inhibition, reversals of ATP and citrate effects in response to AMP activation, and synergism of citrate and ATP inhibitions were also seen to be decreased considerably with muscle PFK of old in comparison to that of young rats. Such age-related changes in muscle PFK suggest the alterations in allosteric regulation of this enzyme during aging of the animal. PMID- 2976265 TI - Autoimmunity and T-cell subpopulations in old age. AB - To investigate the interrelationship between T-cell-dependent immune functions and autoimmune phenomena in old age we determined T-cell subpopulations in 20 aged healthy individuals (80-96 years old) using monoclonal antibodies. These persons were also investigated as to humoral parameters such as antinuclear antibodies, rheumatoid factors (IgG-, IgA-, IgM-RF), antibodies to collagen types I-IV as well as autoantibodies to organ-specific antigens. In addition, immune complexes were determined. We found that aged individuals have an increased frequency of autoantibodies as compared to a young control population, each aged subject presenting with at least one autoantibody species. Immune complexes, however, were only rarely detected. Three individuals showed a slightly increased T-helper/T-suppressor cell ratio, four had a decreased ratio. An increased number of T-suppressor cells was significantly correlated with a lowered incidence of anticollagen antibodies. Other parameters tested by us: fibronectin, laminin, procollagen type III, C3 and C4 complement components, immunoglobulins and acid alpha 1-glycoprotein. Aged individuals have significantly higher serum levels of fibronectin, while laminin and procollagen concentrations are in the normal range. A large percentage of old individuals had increased serum levels of C3 and/or C4. The acute phase protein orosomucoid, however, was in the normal range. PMID- 2976267 TI - [Doppler echocardiography of ventricular diastole]. AB - Non-invasive studies of left ventricular relaxation and filling by means of doppler-echocardiography are of considerable interest owing to easy recording and good reproducibility. However, such physiological parameters as site of measurement, heart rate and, chiefly, age may interfere with the curves obtained. The interpretation of tracings must also take into account the presence of mitral valve pathology (stenosis or regurgitation), aortic stenosis, disorders of atrioventricular (prolonged PR complex) or intraventricular (left bundle branch block) conduction and medication (e.g. vasodilators). In clinical practice, one of the main advantages of the technique is that it makes it possible to distinguish between the physiological left ventricular hypertrophy (LVH) of top class sportsmen (without impaired ventricular filling) and the pathological LVH of hypertrophic cardiomyopathy. The purpose of this study, with a review of the literature, was to take stock of the value and limitations of the doppler indices. PMID- 2976266 TI - [Unstable angina in patients over 70 years of age]. AB - Between October, 1985 and June, 1987, 125 patients aged 70 years or more were admitted to the Cardiology Department of Purpan Hospital, Toulouse for unstable angina pectoris. 107 of these patients have been followed up, i.e.: 76 men and 31 women aged from 70 to 85 years (mean 74.5 +/- 3.2 years). 49 patients (46 p. 100) had angina at rest of the intermediate syndrome type; 46 (43 p. 100) had rapidly progressive effort angina, and 12 (11 p. 100) had effort angina de novo. The patients' general condition was preserved in 85 p. 100 of the cases and altered in 15 p. 100. ECG was normal in only 10 p. 100 of the patients; it showed signs of established ischaemia in 70 p. 100 of the cases and sequelae of infarction in 23 p. 100. The mean cardiothoracic ration was 49.9 +/- 4.8 p. 100. The ejection fraction was 62 +/- 14.6 p. 100, the end-diastolic volume 86.9 +/- 32.2 ml/m2 and the left ventricular end-diastolic pressure 15.8 +/- 7.2 mmHg. 69 patients (64.5 p. 100) had calcifications in their coronary arteries. At coronary arteriography 21 patients (19.6 p. 100) showed stenosis of the main stem, 11 patients (10.3 p. 100) had a single vessel disease, 28 (26.2 p. 100) a two-vessel disease and 47 (43.9 p. 100) a three-vessel disease. Treatment was medical in 41 patients (38.3 p. 100) and surgical in 44 patients (41.12 p. 100); transluminal coronary angioplasty (TCA) was performed in 23 patients (21.5 p. 100).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976269 TI - [Clinical trial with streptokinase and sequential procedures for the treatment of acute myocardial infarction]. PMID- 2976268 TI - [Myocardial infarction caused by the thrombosis of the common trunk of the left coronary artery. Fatal outcome despite an early obstruction removal by angioplasty. Apropos of a case]. AB - We report an exceptional case of acute myocardial infarction due to thrombosis of the left main coronary artery in a 39-year old male patient. After failure of recanalization by systemic fibrinolysis, percutaneous angioplasty under circulatory assistance was successfully performed pending a possible transplantation. The patient unfortunately died of haemodynamic disturbances 48 hours after the infarction. PMID- 2976270 TI - [Refractory unstable angina and coronary angioplasty]. PMID- 2976271 TI - Quality assurance in residences for people with a disability. AB - OBJECTIVE: To review the quality of service in 7 residences for people with a disability administered by the NSW Department of Health in the New England Region. METHOD: Evaluation of each residence by an evaluation team using an evaluation package incorporating three approaches to assessment: process measures, goal-oriented outcome measures, and interviews or questionnaires. RESULTS: This comprehensive review of the 7 residences showed that high standards had been set throughout the New England Region and that the evaluation package was a powerful tool for staff keen to improve the lifestyle of people with a disability. Some of the notable changes made as a direct result of the evaluation included additional staff for recreation and leisure; more flexible administrative control from local hospitals; increased staff consistency in home routine training; and increased community integration programming. PMID- 2976273 TI - The impact of an instant pregnancy test kit on the operations of a major hospital casualty department. AB - The records of all patients on whom a casualty department pregnancy test was performed during the first 4 months of 1986 were retrospectively examined and compared with the records of all patients who in the first 4 months of 1987 had had an instant pregnancy test performed in the same casualty department, to determine the impact of such a kit on diagnostic accuracy, operative procedures and related economic factors. It was demonstrated that instant pregnancy testing significantly improved the accuracy of provisional diagnoses and appeared to help in reducing the number of surgical procedures performed. Furthermore, it was shown that a combination of careful clinical assessment plus the appropriate application of an instant pregnancy test kit could result in a cost saving to the hospital of over $41,000 per annum. PMID- 2976272 TI - Hysterosalpingography in infertility--an experience of 3,631 examinations. PMID- 2976274 TI - Treatment and prophylaxis of Pneumocystis carinii pneumonia. PMID- 2976275 TI - [Quantitative study of the "T" subpopulations in subjects with bronchiolitis]. AB - T-lymphocyte subsets were determined by monoclonal antibodies in eighty children with acute bronchiolitis, eighty healthy controls and 37 patients with respiratory tract infections other than bronchiolitis, all matched by age and sex. T3 and T8-positive cells, but not T3-positive cells, were significantly decreased in the bronchiolitis patients (p less than 0.001), with a subsequent increase in the T-helper/suppressor ratio (p less than 0.001). The decrease in T suppressor lymphocytes could play a role in the pathogenesis of acute bronchiolitis, allowing IgE hyperproduction and alveolar mast cell activation, due to the imbalance in helper/suppressor activity. PMID- 2976276 TI - Taxonomy of family life styles: III. Replication with families with severely mentally retarded children. AB - The home environment and behavior of 97 families with severely mentally retarded children was examined in order to determine the presence of family types. Cluster analysis was performed on 16 measures of proximal home environment. Five unique family clusters were identified: cohesive, control-oriented, responsive-to-child, moral-religious-oriented, nonachievement-oriented, and achievement-oriented. Comparison among the clusters on other family and child characteristics and behaviors revealed significant differences. The family types identified in this study were similar to previously determined family types. These findings appear to support the establishment of a standard classification of families. PMID- 2976277 TI - Coronary artery disease in aortic surgery. AB - The incidence of coronary artery disease in patients coming to aortic surgery and the impact of aggressive preoperative cardiac catheterization and myocardial revascularization was prospectively analyzed in 59 patients. Seventy-five percent of patients had at least one-vessel involvement, and 32% had three-vessel or left main involvement. Patients with electrocardiographic evidence of coronary artery disease had at least one-vessel involvement 84% of the time and three-vessel, left main involvement 36% of the time. Sixty-four percent of patients with no preoperative indications of coronary artery disease had at least one-vessel involvement and 29% had three-vessel, left main involvement. Resting (39 patients) and exercise multiple-gated acquisition scans (22 patients) did not predict the presence of coronary artery disease in patients without a history or electrocardiographic evidence of coronary artery disease. Myocardial revascularization was performed prior to aortic surgery in 17 patients (29%). The operative mortality was 3.7% with two patients dying from noncardiac-related complications. There were two additional deaths prior to aortic surgery with one patient dying during coronary artery bypass grafting, and one dying of aneurysm rupture prior to repair, making the overall mortality associated with this approach 7.4%. Preoperative cardiac catheterization and an aggressive approach toward coronary artery bypass grafting reduces the risk of cardiac complications in aortic surgery. PMID- 2976278 TI - Femorofemoral bypass through the perineum for infection complicating arterial revascularization of the lower limb. AB - Of 587 prosthetic arterial constructions performed between 1980 and 1984 on the aorta and lower limbs, 25 patients were operated on for sepsis. A total of 37 operations were performed, six of which were femorofemoral crossover bypasses through the perineum. Prosthetic material was used in one case and vein in five. The host vessel was the contralateral iliac or femoral artery, the contralateral limb of an aortobifemoral bypass, or the contralateral limb of an axillofemoral bypass in two cases each. The recipient vessel was the profunda femoris artery in four cases, the popliteal artery in one case, and the profunda femoris and popliteal arteries sequentially in one case. Indications for perineal bypass included an infected pseudoaneurysm in the femoral triangle following a femoropopliteal (one case) or aortofemoral bypass (five cases). In two instances, sepsis was bilateral and also required an axillofemoral bypass. Excepting one postoperative death, early results were satisfactory as infection disappeared and arterial reconstructions remained patent in all other cases. This technique should have its place in the armamentarium of extraanatomic bypasses along with other techniques such as the axillofemoral, prepublic femorofemoral crossover, and obturator bypasses. The best indication for perineal bypass is when vascularization of a lower limb from the contralateral side is necessary because of infection in the femoral triangle. PMID- 2976279 TI - Superimposing noise linearizes the responses of primary muscle spindle afferents to sinusoidal muscle stretch. AB - Experiments were conducted in anaesthetized and spinalized cats to measure the extent to which the non-linear response of Ia afferent fibers to sinusoidal muscle stretch as expressed by the peristimulus-time-histograms, PSTHs, can be transformed into a linear one by means of the superposition of random stretch ("mechanical noise"). The gastrocnemius muscles of one hind leg were stretched and the response to sinewave muscle stretch (amplitudes between 0.01 and 4.0 mm, frequencies between 0.1 and 20 Hz) were investigated while band-limited mechanical noise was superimposed on the sinewave stretch. The random stretch upper cut-off frequency was varied between 60 and 300 Hz; the displacements were normally distributed. The noise amplitude sigma, i.e. the standard deviation of the displacement distributions, was varied systematically between 0.002 and 0.4 mm. Mechanical noise was very effective in raising the mean discharge rate. Added to the sinusoidal stretch it prevented the cessation of firing during the release phase of the stretch cycle, or at least reduced the duration of discharge pauses, i.e., a linearization occurred. In general, the larger the noise amplitude, the more the amplitude of the fundamental harmonic component was attenuated and the phase lead reduced. Apart from this rule the particular combination of superimposing small noise (sigma less than 0.02 mm) on small sinewave stretch (A less than 0.02 mm) could enhance the depth of sinusoidal modulation of cycle histograms (compared with responses to pure sinusoids). Linearizing the sinewave response by additional noise allowed the estimation of frequency response characteristics in the otherwise non-linear range of amplitudes (sinewave amplitude 0.5-1.0 mm).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976280 TI - The synthetic retinoid fenretinide does not affect circulating hormone concentrations. PMID- 2976281 TI - Biosynthesis of bloodgroup I and i antigens. A sensitive and specific assay of UDP-GlcNAc:beta-galactoside beta 1----3-N-acetylglucosaminyltransferase activity in hematopoietic cells by HPLC. AB - A sensitive HPLC method for the assay of UDP-GlcNAc:beta-galactoside beta 1----3 N-acetylglucosaminyltransferase activity was developed. Using lactose as an acceptor, the formation of the product GlcNAc beta 1----3Gal beta 1----4Glc can be determined without interference by substrates resulting from enzymatic and chemical breakdown of the donor substrate UDP-GlcNAc. The method is very specific since products of other transferase reactions, which potentially may be formed in the incubations in vitro, elute at positions different from that of GlcNAc beta 1 ---3Gal beta 1----4Glc. By use of this assay method it could be demonstrated that normal and malignant hematopoietic cells and cell-lines, with the exception of erythrocytes and reticulocytes, contain beta 1----3-N acetylglucosaminyltransferase activity. PMID- 2976282 TI - Interaction of biological membranes with the cytoskeletal framework of living cells. AB - The review is focused on the molecular structure and function of the proteins composing the actin-based cytoskeletal cortex, located at the cytoplasmic face of plasma membranes of eucaryotic cells, which stabilizes integral membrane proteins in separate domains of cell membranes. It includes a survey of the molecular properties of the proteins of the erythrocyte membrane skeleton such as spectrin, ankyrin, protein 4.1, and adducin. The properties of the immunological counterparts of erythroid cortical proteins found in nonerythroid tissues and cells are compared. The structural organization and function of the newly discovered class of calcium-binding proteins, nonerythroid peripheral membrane proteins, calpactins, are also described. Finally, the discussion of some experimental models illustrates that the membrane skeleton of living cells is actively involved in a wide variety of essential biological functions ranging from differentiation, to maintenance of cell polarity and cell shape, and regulation of exocytotic processes. PMID- 2976283 TI - A Mg2+-independent Ca2+-stimulated ATPase activity in the tegument of Schistosoma mansoni. AB - A tegumental fraction was prepared from Schistosoma mansoni. This fraction exhibited ATPase activity stimulated by Ca2+ in the absence of Mg2+. The Mg2+ independency was assessed by lowering contaminant Mg2+ using CDTA. The peak of activity was 220 mumol Pi mg-1 protein h-1 and the K0.5 for CaATP was 0.32 mM; the same K0.5 was obtained using MgATP as substrate, in the absence of Ca2+. Both activities may be promoted by the same enzyme since the addition of Ca2+ did not increase the ATPase activity measured in the presence of a saturating MgATP concentration. PMID- 2976284 TI - Effects of castration and antiandrogens (RU 23908 and cyproterone acetate) on glandulain activity and on the secretory epithelium of the rat submandibular gland. AB - 1. The influence of hormones on the submandibular gland of rodents has attracted more attention since the observation of the sexual dimorphism of these organs. Androgens enhance both the development and the secretory activity of the gland. 2. In the present investigation we have studied the differences in wet weight, protein content, glandulain activity and morphometry of granular convoluted tubules of submandibular glands excised from control, castrated and antiandrogen treated (RU 23908 or cyproterone acetate) male adult albino rats. 3. Castration and antiandrogenic treatment did not affect the wet weight or protein content of the organs. Castration or treatment with RU 23908, but not treatment with cyproterone acetate, decreased glandulain activity and the height of the secretory epithelium of granular ducts. 4. The morphology and glandulain activity of submandibular gland granular ducts do not seem to be related solely to plasma testosterone concentration. Different hormonal treatments are needed to identify other factors implicated in the phenomenon. PMID- 2976285 TI - Right ventricular contractility depression in chronically infarcted rats. AB - We studied the in vitro mechanical performance of right ventricular strips obtained from male albino rats submitted to left coronary ligation for 30 days. The infarcted rats developed significant right ventricle hypertrophy together with a reduction of the isometric tension when compared to controls. The reduction was maintained even under positive inotropic intervention, i.e. the cumulative dose-response curves to calcium and adrenaline were displaced downward in parallel in the hypertrophied muscles. We conclude that after 30 days of left ventricle infarction the hypertrophied right ventricular myocardium presents a significant reduction of contractility. PMID- 2976286 TI - Left ventricular hypertrophy differences between male and female renovascular hypertensive rats. AB - In a previous study we demonstrated ventricular hypertrophy in male but not in female sinoaortic denervated rats. To evaluate a possible sexual influence on ventricular hypertrophy of other models for experimental hypertension, we studied two-kidney one clip and one-kidney one clip Goldblatt hypertension. The results showed left ventricular hypertrophy in male but not in female two-kidney one clip groups, despite the same arterial hypertension level in both groups. In the one kidney one clip groups, left ventricular hypertrophy was greater in male than in female rats. The results indicate a sexual influence in ventricular hypertrophy when the arterial renovascular hypertension level is moderate. PMID- 2976287 TI - Evidence for a role of beta-endorphin in activity of nigrostriatal neurons in the rat. AB - The effects of icv administration of beta-endorphin on secretory activity of dopaminergic neurons is described. Homovanillic and dihydroxyphenyl acetic acid levels in cerebrospinal fluid and extracts of brain tissue were determined after administration of beta-endorphin to animals pretreated or not with naloxone. The results suggest that beta-endorphin interferes with formation of dopaminergic metabolites by acting on opioid receptors. PMID- 2976289 TI - Inhibition of field stimulation-evoked relaxations in rat oesophageal smooth muscle by the calcium antagonist PN 200-110. AB - 1. The inhibitory effects of the 1,4-dihydropyridine calcium channel antagonist, PN 200-110 (isradipine), on field stimulation-evoked tetrodotoxin (TTX)-sensitive and -insensitive relaxations were studied in rat oesophageal smooth muscle of the tunica muscularis mucosae. 2. The TTX-insensitive relaxation was inhibited by PN 200-110 in a stereoselective manner with the (+)-(S)-isomer displaying a 1000 fold greater inhibitory potency than the (--)-(R) isomer. A similar potency was noted for inhibition of high K+ -evoked contractions. 3. TTX-sensitive relaxations evoked by field stimulation and contractions elicited by the muscarinic cholinoceptor agonist, cis-2-methyl-4-dimethylamino-methyl-1,3 dioxolane methiodide (cisdioxolane) were considerably less sensitive to inhibition by PN 200-110, although, again, stereoselectivity for PN 200-110 was apparent. 4. Pretreatment with (+)-(S)-PN 200-110 resulted in a non-competitive displacement of the Ca2+ concentration-response curves obtained in the presence of either isotonic 50 mM KCl or cisdioxolane. The effect of K+ was 10 fold more sensitive than that of cis-dioxolane. 5. The potency rank orders for inhibition of TTX-insensitive field stimulation-evoked relaxations and K+ -mediated contractions in a series of calcium channel antagonists were closely correlated; (+)-(S)-PN 200-110 showing highest potency followed by nifedipine, verapamil, diltiazem, (--)-(R)-PN 200-110. 6. It is concluded that TTX-insensitive relaxations are dependent upon an influx of extracellular Ca2+ through potential operated calcium channels. PMID- 2976290 TI - [Early embryo development of the bird egg subjected to a magnetic field]. AB - Chick and quail eggs exposed during the first 4 or 7 days of incubation to 50 Hertz and 5 Gauss sinusoidal magnetic fields developed normally. The percentages of dead and abnormal embryos observed in such conditions are identical to those arising spontaneously in control eggs. PMID- 2976288 TI - Bisoprolol improves perfusion of ischaemic myocardium in anaesthetized pigs. AB - 1. The ability of the cardioselective beta-adrenoceptor antagonist bisoprolol ((+/-)-1-[4-(2-isopropoxyethoxymethyl)-phenoxy]-3-isopropyl-amino -2-propanol hemifumarate, EMD 33512) to suppress isoprenaline-induced increases in heart rate and maximal rate of rise in left ventricular pressure (LVdP/dtmax) was studied in 6 anaesthetized pigs given 4 cumulative doses (16, 64, 256 and 1024 micrograms kg 1). Bisoprolol was about 2 times more effective in suppressing isoprenaline induced increases in LVdP/dtmax than those in heart rate. 2. In 8 animals which had a partial stenosis of the left anterior descending coronary artery (LADCA), the effects of 3 consecutive doses (50, 200 and 750 micrograms kg-1) of bisoprolol were studied on systemic haemodynamics, regional myocardial perfusion and function. The effects of the drug were compared with those obtained in a group of 9 animals with LADCA stenosis which did not receive any treatment. 3. The lowest dose of bisoprolol (50 micrograms kg-1) increased perfusion of the ischaemic myocardium (which had been reduced from 123 +/- 20 ml min-1 100 g-1 to 42 +/- 11 ml min-1 100 g-1) by 21 +/- 10 ml min-1 100 g-1 (P less than 0.05). In particular the subendocardial layers, which were most severely affected by the stenosis (a decrease from 128 +/- 19 ml min-1 100 g-1 to 20 +/- 6 ml min-1 100 g 1) benefited from the administration of the drug (an increase of 30 +/- 10 ml min 1 100 g-1). Perfusion of the subepicardium was not significantly affected. With the higher dose only a minor additional improvement in perfusion of the ischaemic myocardium was observed. 4. The negative chronotropic response is the most likely factor leading to the improvement in perfusion. 5. Myocardial wall thickening, which decreased from 41 +/- 2% to 9 +/- 4% (P less than 0.05) due to the hypoperfusion, did not improve after administration of the drug. This lack of improvement may possibly be due to the duration of ischaemia before and the magnitude of the flow deficit after bisoprolol administration. 6. Between 15 and 60 min of ischaemia, 5 of the 9 untreated animals had an episode of ventricular fibrillation compared with only 1 of the 8 animals treated with bisoprolol, in spite of an initially larger flow reduction in the treated animals. The more homogeneous flow distribution after bisoprolol might account for the lower incidence of arrhythmias in this group. 7. It was demonstrated that bisoprolol improves perfusion of ischaemic myocardium in anaesthetized pigs even at doses (50.pgkg-1) that only moderately antagonize isoprenaline-induced cardiostimulatory effects. PMID- 2976291 TI - [Comparative study of the kinetic properties of the neutral maltase of human granulocytes and of the kidney maltase of the rat]. AB - Neutral maltase from human granulocytes has a different substrate specificity from the human neutral maltase of kidney, though it has been reported that these two enzymes are immunologically similar. We report here that human granulocyte neutral maltase is similar to the neutral maltase from rat's kidney as regards the substrate specificity and the inhibition by Tris and maltodextrins. We also report a different thermal stability that might imply some structural differences between the two enzymes. PMID- 2976292 TI - [Effect of natural or synthetic detergents on the transport of D-glucose in the membranes of vesicles of the brush border of the intestine of the rabbit]. AB - We describe here the effects of natural and synthetic detergents on the D-glucose transport into brush-border membranes of vesicles of rabbit's intestine. Two synthetic detergents: Triton X-100 and dodecyltrimethylammonium bromide have been found very strong inhibitors (more than 50 p. 100 of inhibition of maximal D glucose uptake). Kinetic studies showed that these detergents behaved as mixed type inhibitors. The Na+-dependent transport of amino acids (aspartic acid, lysine, phenylalanine) is only poorly affected by dodecyltrimethylammonium bromide, while Triton X-100 inhibits unspecifically all the transport studied. PMID- 2976293 TI - [Inhibition of mitochondrial respiration by mepivacaine]. AB - In the last years our researches on neurotropic drugs follow our hypothesis that the strong effects on nervous system have always hidden more widespread effects on all tissues and cells. It is often required to employ local anesthetics in practising dentistry and orthodontics, particularly when children have to be treated. We have assayed in vitro one of these dental anesthetics, mepivacaine, on liver rat mitochondria: it depresses the respiration coupled to phosphorylation in mitochondria having a good respiratory control; so respiratory control too is depressed, but P/O ratio is unaffected; also respiration uncoupled by 2.4-dinitrophenol is depressed. Depressing respiration cooperates with anesthesia; unchanging P/O is good for the health of the cells and tissues treated by the mepivacaine. PMID- 2976294 TI - [Stimulation of the chemiluminescence of human polymorphonuclear leukocytes in various stages of the intraerythrocyte development of Plasmodium falciparum]. AB - The synchronized cultures of Plasmodium falciparum were used to stimulate in vitro the chemiluminescence of human polymorphonuclear leukocytes in the presence of immune serum. The schizonts were concentrated by Percoll gradient centrifugation method (density 1.085 and osmolarity 285 mOsmol), and placed in culture, treated 6 hours later by sorbitol. Under incubation at constant temperature and pressure, the rate of synchronization reached 85% for schizonts during 5 replicative cycles. Every asexual stages of Plasmodium falciparum were used separately to stimulate polymorphonuclear leukocytes: merozoites were the most effective, followed by schizonts, trophozoites, and lastly supernatants of cultures containing degradation products of parasites. PMID- 2976295 TI - [Flux cytometry of the cellular cycle of leukemic cells of the blood]. AB - The coordination of flux cytometry and of a techniques of leukoconcentration allowed to determine the cellular cycle of nucleated cells of circulating blood, without logs nor enrichment of cellular type on a definitive moment. The study of acute leukemias allow to conclude that: 1) it exists in peripheral blood a synthetic activity of ADN bound to the presence of leukemic or blastic cells; 2) this activity allows to appreciate the spontaneous variations of synthesis and the incidence of chemotherapy. PMID- 2976296 TI - [Experimental conditions of a technic of in vitro transcription using placental nuclei]. AB - To examine the expression of human placental lactogen, we investigated transcription in isolated placental nuclei. Using SH-agarose affinity chromatography, we have isolated Hg-RNA synthetized in vitro. These RNA are translated in a wheat germ cell-free system. hPL synthetized in vitro, accounting for 20% of total protein. This system should prove useful for the study of gene expression of hPL. PMID- 2976297 TI - [Histologic study of the outcome of components of nerves of human corpses grafted in nude mice (a model of chimeric intercellular communication)]. AB - Human corpse nerves are microsurgically grafted in nude mouse and biopsied from 1 to 152 days after graft. Schwann cells ever survived and probably produce angiogenic factors stimulating mice vessels. Others nerves are grafted end to end with mouse sciatic nerve. Mouse's neurites invading grafts are surrounded by human Schwann cells and set up normal chimeric nerve. PMID- 2976298 TI - [Effects of 3,5,3'-triiodo-L-thyronine on the enzymes responsible for the metabolism of xenobiotics in the rat after increase in the dietary supply of cholesterol]. AB - A cholesterol rich diet fed to rats was found to increase the cytochrome P 450 content in hepatic microsomes. Furthermore, the variations of the same parameters promoted by 3,5,3'-triiodo-L-thyronine were strongly reduced in cholesterol supplemented rats. Cholesterol prevented the inducing effects of phenobarbital but did not oppose its decreasing effect on maximal fluorescence of ANS and PNA introduced into the microsome suspensions. PMID- 2976299 TI - [Influence of 3,5,3'-triiodo-L-thyronine on the hepatic enzyme activities responsible for the metabolism of xenobiotics during the development of the chick embryo]. AB - The influence of thyroid hormones on microsomal drug metabolizing enzymes was studied in hypothyroid newborn rats and chick embryos. Administration of 3,5,3' triiodo-L-thyronine strongly decreased the microsomal cytochrome P 450 content in hypothyroid new-born rats and thus could render the rat pup more susceptible to hepatotoxicity from drugs. The drug metabolizing system in 20 days old chick embryos was less sensitive to the effects of thyroid hormone, but administration of phenobarbital was accompanied by a strongly induction effect on microsomal enzyme activities. PMID- 2976301 TI - [Studies on the levels of plasma and platelet beta-thromboglobulin in thrombocytopenia]. PMID- 2976302 TI - [Beta-endorphin like-immunoreactives content changes in the CSF of patients with acute brain injury and its clinical significance]. PMID- 2976300 TI - [Hepatic microsomal delta 9 desaturation of stearic acid in the spontaneously diabetic female BB rat]. AB - delta 9 desaturation of stearic (1-14C) acid has been estimated from incubation of liver microsomes of adult female spontaneously diabetic BB rat, an animal model resembling the spontaneous juvenile diabetes in humans, comparatively to adult female control Wistar rat. The animals were sacrificed, when hyperglycemic, 24 hours after the last insulin injection to the BB rats. Stearic acid delta 9 desaturase activity is drastically depressed in the BB rats when fatty acid composition of liver phospholipids and microsomal total liver lipids are changed in spite of the daily injection of insulin necessary for the BB rats survival. PMID- 2976303 TI - [Myocardial hypertrophy and left ventricular function in hypertensive patients: Doppler echocardiographic evaluation]. PMID- 2976304 TI - Transluminal coronary angioplasty in the treatment of silent ischemia. AB - Fifty-four asymptomatic patients with positive thallium exercise tests underwent coronary angiography followed by coronary angioplasty (PTCA), as the primary therapy for silent ischemia. The procedure was technically successful in 89% of these patients. Emergency bypass graft surgery was necessary in 2 (3.6%) and q wave myocardial infarction occurred in 1 (1.8%) of these. All fifty-four patients have been followed for a mean of 35 months since angioplasty. Of the 48 patients with initially successful PTCA, 12 had either clinical restenosis (9/14 or 19%) or a new lesion (3/48 or 6%) during follow-up, which required a repeat PTCA. At the longest follow-up, 46 (85%) had been successfully treated with on or more PTCA procedures. Two patients (3.6%) had sustained late q-wave myocardial infarction and two additional patients reported angina pectoris. There were no deaths. Angioplasty as a primary therapy for silent ischemia appears efficacious, with success and restenosis rates comparable to those in the symptomatic population. Event-free survival is improved, compared with natural history data for patients with silent ischemia from other studies. Prudent risk/benefit analysis may help to define subgroups most likely to benefit from this intervention. PMID- 2976305 TI - Monitoring of ischemia during percutaneous coronary angioplasty: influence of guidewire, balloon placement, and lead selection. AB - We examined the sensitivity of the surface 12-lead electrocardiogram (ECG) for detecting ischemia during guidewire and deflated balloon passage as well as during balloon inflation in proximal epicardial stenoses during percutaneous transluminal coronary angioplasty (PTCA) of 55 patients. Ischemia (ST change greater than or equal to 0.1 mV) by 12-lead ECG was detected in 28% of patients after guidewire passage, in 50% after deflated balloon passage, and in 76% during balloon inflation vs. 17%, 14%, and 50%, respectively, by limb lead monitoring alone. The best single lead for detecting ischemia during PTCA was V2 for left anterior descending and circumflex and III for right coronary artery inflations. The addition of a selected second precordial lead further enhanced ischemia monitoring. We conclude that ischemia is common during PTCA even during wire and deflated balloon passage, that the 12-lead ECG is more sensitive for monitoring ischemia during PTCA than conventional techniques, and that laboratories can optimize their ability to detect ischemia during PTCA by selecting appropriate leads. PMID- 2976306 TI - Assessment of coronary angioplasty: comparison of visual assessment, hand-held caliper measurement and automated digital quantitation. AB - Digital subtraction coronary angiograms (DSA) of 27 patients who had undergone coronary angioplasty (PTCA) to a total of 32 lesions were analyzed using an automated border-detecting computer program and hand-held caliper measurement of diameter percent stenosis. The results were compared with visual interpretation of the 35-mm cineangiograms. Visual reports significantly overestimated the pre PTCA diameter percent stenosis (P less than .001) and underestimated the residual post-PTCA narrowing compared with the automatic computer program (P less than .001). Caliper measurements overestimated significantly the pre-PTCA stenosis in comparison with the computer (P less than .01), but post-PTCA the two methods did not differ significantly (P = .105). There was a positive but poor correlation between caliper and computer measurements (r = .43, P less than .05) performed on the pre-PTCA digital angiograms. Post-PTCA the two methods correlated better (r = 0.78, P less than .001), but further statistical analysis showed important discrepancies between them. The correlation of visual reports and computer measurements was poor pre-PTCA (Kendall's tau = 0.32, P less than .05) and not statistically significant post-PTCA (tau = 0.64, P = .5). We conclude that there is observer bias in the visual reporting of angioplasty results, so that pre-PTCA lesions are overestimated, whereas post-PTCA are underestimated. Hand-held caliper measurement improves the assessment of coronary stenoses in comparison with the visual report but still may not altogether eliminate observer bias; it may overestimate the pre-PTCA stenoses compared with automated border detection. PMID- 2976307 TI - Brachial artery injury as a complication of cardiac catheterization: percutaneous transluminal angioplasty and streptokinase as a treatment alternative. AB - Brachial artery injuries are a well-described complication of cardiac catheterization. Traditional therapy has involved surgical repair. We report a case treated with balloon angioplasty followed by streptokinase. This procedure may provide a treatment alternative in a select group of patients. PMID- 2976308 TI - Simultaneous double balloon coronary angioplasty through a single guiding catheter for bifurcation lesions. AB - To enhance the safety, efficacy and expediency of coronary bifurcation lesion angioplasty, we report the use of two dilatation catheters, advanced simultaneously through a single guide catheter in three patients. Successful dilatation was performed in each. The technique involves the use of either two new low-profile dilatation catheters in an 8 French large lumen guide catheter or a single new low-profile dilatation catheter coupled with a conventional over-the wire catheter within a new large lumen 9 French guide. PMID- 2976309 TI - Plasminogen activation: biochemistry, physiology, and therapeutics. AB - The mammalian serine protease zymogen, plasminogen, can be converted into the active enzyme plasmin by vertebrate plasminogen activators urokinase (uPA), tissue plasminogen activator (tPA), factor XII-dependent components, or by bacterial streptokinase. The biochemical properties of the major components of the system, plasminogen/plasmin, plasminogen activators, and inhibitors of the plasminogen activators, are reviewed. The plasmin system has been implicated in a variety of physiological and pathological processes such as fibrinolysis, tissue remodeling, cell migration, inflammation, and tumor invasion and metastasis. A defective plasminogen activator/inhibitor system also has been linked to some thromboembolic complications. Recent studies of the mechanism of fibrinolysis in human plasma suggest that tPA may be the primary initiator and that overall fibrinolytic activity is strongly regulated at the tPA level. A simple model for the initiation and regulation of plasma fibrinolysis based on these studies has been formulated. The plasminogen activators have been used for thrombolytic therapy. Three new thrombolytic agents--tPA, pro-uPA, and acylated streptokinase plasminogen complex--have been found to possess better properties over their predecessors, urokinase and streptokinase. Further improvements of these molecules using genetic and protein engineering tactics are being pursued. PMID- 2976310 TI - [Effect of anisodamine (654-2) on rat neutrophil function]. PMID- 2976311 TI - [Augmentation effect of selenium on IL-2 production of lymphocytes without affecting IL-2 responsiveness]. PMID- 2976312 TI - [Studies on human cervical carcinoma cell line. I. Establishment of cloned human cervical carcinoma cell line and biological characteristics of CC-801]. PMID- 2976313 TI - [Immunoradiometric assay for tumor associated antigen CA-50]. PMID- 2976314 TI - [Biosynthesis of prostaglandin endoperoxide intermediates and its application]. PMID- 2976315 TI - [Effect of sodium and calcium intake on blood pressure, plasma calmodulin and erythrocyte ATPase activities]. PMID- 2976316 TI - [Immuno-pathological studies on muscles of polymyositis and dermatomyositis]. PMID- 2976317 TI - [The crystal structures of harringtonine and homoharringtonine]. PMID- 2976318 TI - [A group of monoclonal antibodies against human fibrinogen and its fragments]. PMID- 2976319 TI - [Measurement of human serum IgE by reverse passive hemagglutination with monoclonal anti-IgE antibodies]. PMID- 2976320 TI - [Studies on attenuation of cardiovascular responses during endotracheal intubation following rapid-sequence induction of anesthesia]. PMID- 2976321 TI - [Purification and completion of LA795 Vv-Vt in vivo-in vitro system]. PMID- 2976322 TI - [Schistosomiasis haematobia (report of 22 cases)]. PMID- 2976323 TI - [Reverse hemagglutination (RHA) test for detection of ovarian cancer antigen in serum]. PMID- 2976324 TI - Characteristics of social interactions between mothers and their disabled infants: a review. AB - This paper examines interactional characteristics of disabled infants and young children and their mothers. Interactional characteristics of disabled infants and young children differ from those of non-disabled children in several ways: disabled children provide fewer and less readable cues to the mother; they demonstrate less positive affect, more negative affect, and more muted affect; and they have more difficulties synchronizing turn-taking than non-handicapped children. Maternal interactions with disabled children differ from those of mothers with non-disabled children in dimensions involving maternal activity and positive affect. It is suggested that the social deficits commonly observed in disabled preschoolers and older children may have begun in the altered social interactional patterns observed in infancy. Various intervention strategies are described which assist parents to achieve more pleasurable, reciprocal, child centered interactions with their disabled infants. PMID- 2976325 TI - Postoperative nausea and vomiting. A comparison of sufentanil, nitrous oxide, and isoflurane. PMID- 2976326 TI - A sensitive non-radioactive assay for pyrimidine nucleoside phosphorylase using reversed-phase high performance liquid chromatography. AB - A new sensitive assay for pyrimidine nucleoside phosphorylase using non radioactive substrates is described. With the natural substrate uridine (UR) and the analog, 5'-deoxy-5-fluorouridine (5'dFUR) conditions have been optimized to measure the product formation with reversed-phase high-performance liquid chromatography. Using automated injection large series of samples may be analyzed. The assay for UR phosphorylase appeared to be comparable to existing methods with radiolabeled UR as substrate regarding sensitivity and linearity. The assay has been used to measure kinetic parameters for 5'dFUR and UR in two cell lines from intestinal origin. PMID- 2976328 TI - New retinoids for dermatologic diseases. Uses and toxicity. AB - Systemic administration of retinoids is frequently associated with mucocutaneous side effects, liver toxicity, and abnormalities of the serum lipid profile. Currently, the main factors limiting more common use of retinoids are their teratogenicity and chronic bone toxicity. Treatment with retinoids requires appropriate selection of patients, careful periodic monitoring of the clinical response, and appropriate laboratory tests. Careful therapeutic management is necessary in order to prevent or minimize side effects. PMID- 2976327 TI - Effect of atrial peptide on collecting duct function. AB - 1. The effects of synthetic rat atrial natriuretic peptides (ANP) on diffusional 22Na+, 36Cl- and tritiated water (THO) permeability of in vitro microperfused rat papillary collecting ducts and the effect in vivo of ANP on stop-flow sodium concentrations in the terminal segment of rabbit nephrons were studied. 2. The addition of 4 x 10(-8) or 4 x 10(-7) mol/l ANP to the medium or perfusion solution did not alter diffusional 22Na+ or 36Cl- permeability of microperfused rat papillary collecting ducts. 3. The basal diffusional THO permeability of papillary collecting ducts was not altered when 4 x 10(-7) mol/l ANP was present in the medium and did not inhibit the increment in diffusional THO permeability induced by vasopressin or reduce the permeability to water in a duct previously stimulated by vasopressin. 4. The administration of ANP (2 micrograms/kg bodyweight) to rabbits in water diuresis did not alter systemic blood pressure but induced a marked natriuresis and increases in urine flow and potassium excretion. This natriuresis was not associated with alterations in stop-flow sodium reabsorptive capacity or sodium permeability of the collecting tubules and ducts. 5. Previously reported in vivo clearance data suggest that ANP causes, at least in part, a natriuresis by altering sodium transport in the medullary collecting ducts. In this study, however, a direct effect could not be demonstrated and it is possible that the medulla needs to be functioning in its normal environment for such effects to be demonstrated. PMID- 2976329 TI - Percutaneous transluminal coronary angioplasty in multivessel coronary disease patients: short- and long-term follow-up in single and multiple dilatations. AB - Transluminal coronary angioplasty was successfully performed in 658 of 752 patients with multivessel disease. An angiographic success was achieved in 1198 of 1358 lesions (88%). One lesion was attempted in 338 patients (45%); 2 in 273 (37%); 3, in 101 (13%); and, 4 or more in 40 cases (5.3%). Significant complications occurred in 39 patients (5.2%): 19 (2.5%) had a transmural infarction; 26 (3.5%) required urgent myocardial revascularization; and 14 (1.9%) died. An apparent lesion recurrence occurred in 233 of 658 (35%) patients with 162 of 171 (95%) having a successful second coronary angioplasty. A second apparent lesion recurrence occurred in 37 of 162 patients (23%) with 24 of 28 (86%) having a successful third coronary angioplasty. Clinical improvement (mean follow-up: 31 +/- 17 months) persisted in 81% of successful patients. The cumulative probability of survival was 91.5% at 72 months. Survival was adversely affected, at 63 months, by the presence of prior bypass surgery (no prior bypass surgery, 94% vs. prior bypass surgery, 86%; p less than 0.05): at 24 months by a low left ventricular ejection fraction (less than or equal to 35%, 82% vs. left ventricular ejection fraction greater than 35%, 95%; p less than 0.01) and, at 57 months, in the multiple dilatation group with prior bypass surgery (no bypass surgery 96% vs. prior bypass surgery 84%; p less than 0.05). Multiple dilatation had a beneficial effect upon survival, at 27 months, in patients with a left ventricular ejection fraction less than or equal to 35% [single dilatation, 74% vs. multiple dilatation, 93%; p less than 0.001], and in patients greater than or equal to 70 years, at 39 months (79% vs. multiple dilatation, 92%; p less than 0.01). These data suggest that coronary angioplasty can be an effective treatment in patients with multivessel coronary disease without the need to dilate all diseased vessels, with good success, acceptable complication rates, and a reasonable expectation of satisfactory long-term clinical improvement. PMID- 2976330 TI - ST-segment depression in inferior ECG leads during percutaneous transluminal coronary angioplasty for left anterior descending artery. AB - The mechanism of inferior ST depression during percutaneous transluminal coronary angioplasty (PTCA) for the left anterior descending artery (LAD) was studied in 108 patients with isolated LAD lesion. In 49 patients (Group I) ST depression in inferior leads was observed, and 59 patients (Group II) showed no inferior ST depression. In the lateral lead (I or a VL), the incidence (43 cases; 88% vs. 5 cases; 9%) and degree (1.5 +/- 0.8 mm vs. 0.2 +/- 0.4) of ST elevation were significantly greater in Group I than in Group II. There was a significant inverse correlation (r = -0.57, p less than 0.01) between ST depression in the inferior lead and ST elevation in the lateral lead, but no correlation was found between anterior leads (V2-4) and the inferior leads. Elevation of diastolic pressure of the pulmonary artery (8.7 +/- 4.8 vs. 5.7 +/- 2.8 mmHg) and lowering of systolic pressure of the femoral artery (-25.7 +/- 24.3 vs. -11.8 +/- 8.3 mmHg) were significantly greater in Group I. These findings suggest that ST depression in the inferior leads during LAD angioplasty was due to a purely electrical phenomenon and not to the concomitant ischemia of the inferior wall. However, this change was more frequently seen in patients with larger myocardial ischemia and, presumably, was more often related to ST elevation in the lateral lead, which is reciprocal in position. PMID- 2976331 TI - Hyperresponsiveness to water immersion in sodium retaining cirrhotics: the role of atrial natriuretic factor. AB - Cirrhotics do not respond uniformly to head-out water immersion (HWI). Some cirrhotics have an exaggerated natriuresis while others are unresponsive. We studied the humoral and urinary responses to HWI in 5 cirrhotics and compared this to 5 normals on the same 20 mM sodium intake. These cirrhotics had a documented tendency to salt and water retention but had minimal or no evidence of significant ascites. Plasma levels of atrial natriuretic factor (ANF) increased during immersion in both groups. However, the rise in plasma ANF from baseline during each hour of immersion was significantly greater in the cirrhotics (p less than 0.01), than in the normal volunteers. In these cirrhotics, HWI not only corrected the pre-immersion tendency to sodium retention, but by 3 hours had produced an exaggerated natriuretic response (p less than 0.05). It is possible that the greater risk of ANF in cirrhotics is responsible for this natriuretic response. However the reason for the larger rise in cirrhotics is unclear; presumably either a greater central redistribution of plasma volume occurs in cirrhotics following immersion, or there may be greater sensitivity of the ANF release system in cirrhotics. PMID- 2976332 TI - Fate of polyester arterial prostheses implanted as thoraco-abdominal by-passes in dogs: haematology, pathology, and biochemistry. AB - This study was designed to evaluate the haematological, pathological and biochemical characteristics of the flow surface in polyester arterial prostheses. Their implantations as thoraco-abdominal by-passes in dogs were scheduled to remain in place for periods ranging from four hours to six months. The haematologic parameters of the hosts were evaluated before implanting the prostheses and before harvesting the graft. To quantify the thrombogenicity of the flow surface, the platelet deposition and fibrinogen uptake were measured by means of an isotopic technique involving Indium-III labelled platelets and iodine 125 labelled fibrinogen. Synthesis of prostacyclin (PGI2) and thromboxane (TXA2) by the internal capsulae was determined by radioimmunoassay. The pathology was assessed according to previously described protocols. The haematologic parameters of the hosts did not alter substantially in the presence of the prostheses. The observed changes, though slight, were not pathologically significant 48 hours after implantation. Platelet and fibrin deposition was highest on the luminal surface of the prostheses 24 hours post-implantation. These decreased and became negligible after one month when the flow surface became smooth and glistening. The PGI2/TXA2 ratio of the luminal surface was much lower than that observed in native aortic tissue. These results show that the large calibre Dacron vascular prosthesis has a flow surface with a low degree of blood compatibility. The concentration of PGI2 on the newly-formed luminal surface remained low. This characteristic and the incomplete endothelialization of the luminal surface might be among the main factors that determine the high failure rate of synthetic prostheses when used as small diameter blood conduits. PMID- 2976333 TI - Physiological effects of synthetic atrial natriuretic factor in normal conscious dogs. AB - The objective of this study was to examine the integrative physiologic effects of atrial natriuretic factor (ANF). Synthetic 99-126 ANF was administered to 6 normal conscious dogs as two consecutive infusions (0.02 and 0.1 microgram/kg/min respectively) each over 30 min: each infusion was preceded by a priming dose of 1 microgram/kg. With the first infusion, mean arterial pressure declined from 113 +/- 2 to 103 +/- 4 mmHg, pulmonary capillary wedge pressure declined from 9.6 +/- 0.5 to 7.2 +/- 0.7 mmHg and right atrial pressure declined from 9.8 +/- 0.4 to 8.0 +/- 0.7 mmHg (all p less than 0.05). No change in heart rate was seen. Despite very high plasma ANF concentrations, no further decline in arterial or central filling pressures were seen in the second infusion. In contrast, cardiac output declined progressively from 3.8 +/- 0.2 to 2.8 +/- 0.21/min (p less than 0.01) by the end of second infusion. Plasma renin activity declined from 2.2 +/- 0.7 to 0.9 +/- 0.3 ng/ml/hr (p less than 0.05) while plasma norepinephrine remained unchanged. Urine output and sodium excretion increased in a dose dependent manner. The diverse time course of the hemodynamic, renal and neuroendocrine effects suggests these effects of ANF are mediated by different mechanisms. Furthermore, the failure of heart rate and plasma norepinephrine to increase despite a significant decline in blood pressure and cardiac output suggests ANF may have a depressant effect on the sympathetic nervous system. PMID- 2976334 TI - Procainamide in vivo modulates suppressor T lymphocyte activity. AB - Autoantibodies to histone and denatured DNA have been found in 80% of patients treated with procainamide. Of these 10 to 20% will eventually develop a Systemic Lupus Erythematosus-like syndrome. Although the mechanism by which procainamide exerts its effect is unknown, in vitro studies suggest that procainamide may inhibit suppressor T cell activity. We have studied the immune function of 18 patients receiving a two hour infusion of procainamide during transvenous catheter electrophysiologic studies. There was no difference between pre and post infusion samples with respect to T and B cell mitogenesis or pokeweed mitogen induced immunoglobulin secretion. However, in seventeen of eighteen patients, there was a marked decrease in Concanavalin A-inducible suppressor cell activity. This decrease appeared to be related to the amount of procainamide infused as high dose samples showed less suppressor activity than low dose samples. Thus the data show that procainamide, when given in vivo, leads to a rapid and dose dependent decrease in suppressor cell activity. PMID- 2976335 TI - Effect of atrial peptide on gastric acid secretion in rats. AB - The effect of synthetic rat atriopeptin (AP) II was examined on basal, vagally and carbachol-induced gastric acid secretion in anesthetized rats. AP II infusion, at stepwise increasing doses of 2, 20 and 100 ng/kg/min, had no effect on basal acid secretion. At doses of 2 and 20 ng/kg/min, AP II augmented vagally induced acid secretion significantly. The secretory response to vagal stimulation + AP II 20 ng/kg/min was completely abolished by atropine. In contrast a higher dose of AP II (50 ng/kg/h) reduced vagally induced acid secretion significantly. This dose of AP II also reduced acid secretion during direct cholinergic stimulation by carbachol, while the lower dose of 20 ng/kg/min had no effect on carbachol-induced acid secretion. The present data demonstrate for the first time an effect of atrial peptide on gastric acid secretion. At lower doses AP II augments the vagal influence on parietal cell function perhaps by augmenting vagally induced acetylcholine release. At higher doses AP II exerts an inhibitory effect on parietal cell function during vagally and carbachol-induced acid secretion, suggesting different and as yet unknown mechanisms of action. These results raise the possibility that the heart can exert a hormonally mediated influence on the regulation of gastric acid secretion. PMID- 2976337 TI - [Carbocalcitonin in the treatment of senile osteoporosis and hypercalcemic syndromes]. PMID- 2976336 TI - Ex vivo platelet studies following oral nisoldipine in normotensive insulin dependent diabetics and non-diabetic controls. AB - The effect of 24 hours and 7 days treatment with nisoldipine (10 mg, twice daily) on platelet function was studied in 12 normotensive volunteers of whom six were insulin-dependent diabetics without clinical evidence of vascular complications. Platelet aggregation was assessed by platelet rich plasma (PRP) and whole blood (WB) techniques. In addition, the effect of nisoldipine on platelet hyperaggregability following exercise was assessed. After taking nisoldipine for 24 hours, in vitro platelet hypersensitivity to adenosine diphosphate was observed in PRP (p less than 0.01) and WB (p less than 0.01), to adrenaline in WB (p less than 0.03), and to collagen in PRP (p less than 0.02). After seven days treatment, platelet sensitivities to all agonists at rest in both PRP and WB showed no differences from pre-treatment values. Exercise-induced platelet hypersensitivity in WB to all three agonists was unchanged after nisoldipine treatment. Plasma noradrenaline and adrenaline concentrations increased after 24 hours treatment, although changes in agonist EC50s at 24 hours were not related to changes in plasma catecholamine levels. No effects of nisoldipine were observed on platelet thromboxane B2 release in PRP, or on plasma beta thromboglobulin levels. No differences in the effects of nisoldipine were observed between diabetic and non-diabetic subjects. Nisoldipine treatment for seven days is not associated with altered platelet function, but platelet hypersensitivity is observed after treatment for 24 hours in both insulin dependent diabetics and controls. PMID- 2976338 TI - [Effects of the treatment with ticlopidine in hemodialyzed patients]. PMID- 2976339 TI - [Drug addiction and HIV infection. Changes in immunohematological parameters and clinical picture in a cohort of drug addicts after 1-year follow-up]. PMID- 2976340 TI - [Efficacy and tolerance of intravenously administered epomediol in patients with hepatopathy]. PMID- 2976341 TI - [Severe proarrhythmic effects of flecainide]. PMID- 2976342 TI - [Treatment of chronic cardiac decompensation in patients 70 years of age and older]. PMID- 2976343 TI - [Disorders of the water-electrolyte balance. Clinical, diagnostic and therapeutic aspects]. PMID- 2976344 TI - [Hepatocholecystopathy]. PMID- 2976345 TI - [Famotidine in the therapy and prevention of duodenal ulcer]. PMID- 2976346 TI - [Sideropenia: modality and length of therapy]. PMID- 2976347 TI - [Changes in metabolic control after interruption of oral hypoglycemic agents in type II diabetic patients with secondary failure during combined therapy]. PMID- 2976348 TI - [Interferon in the treatment of non-Hodgkin's lymphoma]. PMID- 2976349 TI - [Gastroesophageal reflux. Physiopathology, instrumental and clinical diagnosis and therapy]. PMID- 2976350 TI - [Left side mass (pseudosplenomegaly) caused by a large retroperitoneal ganglioneuroma]. PMID- 2976351 TI - [Macrolides: a critical re-evaluation]. PMID- 2976352 TI - [Effects of nicardipine in the treatment of essential arterial hypertension]. PMID- 2976353 TI - [Naltrexone, opiate antagonist, in the therapy of drug dependence]. PMID- 2976354 TI - [Effects of medium- and long-term treatment with nicardipine on arterial blood pressure, hemodynamics and renal function in patients with arterial hypertension of mild and moderate degree]. PMID- 2976356 TI - [Neoplastic hypercalcemia]. PMID- 2976355 TI - [Treatment of postoperative urinary infections in obstetrics and gynecology with cinoxacin]. PMID- 2976357 TI - [Lipid peroxidation of mononuclear cells of peripheral blood before and after therapy with vitamin E]. PMID- 2976358 TI - [Infectious diarrhea. Etiopathogenetic, clinical and therapeutic aspects]. PMID- 2976359 TI - [Magnetic resonance in the evaluation of patients with lymphoma]. PMID- 2976360 TI - [Quality of life]. PMID- 2976361 TI - Health science students' images of disabled people. PMID- 2976362 TI - Carboxyfluorescein distribution in ocular tissues of normal and diabetic rats. AB - Quantitative fluorescence microscopy was used to study carboxyfluorescein distribution across the blood-ocular barrier of control and streptozotocin diabetic rats at 2 min, 1 and 2 hr after dye injection (125 mg/kg iv). Measurement of dye concentrations in plasma, urine and feces demonstrated increased plasma clearance and increased urinary clearance of carboxyfluorescein in diabetic rats. Unbound plasma dye concentration in the diabetic animals fell to 34% of the control level at 1 hr after injection; the corresponding plasma concentration vs. time integral was reduced to only 74% of the control value. The presence of a less fluorescent glucuronide conjugate of carboxyfluorescein was not detected in plasma, urine or feces. Fluorescence intensity in the ocular tissues measured, including choriocapillaris, pigment epithelium, retina, ciliary epithelium, iris, and cornea, was not higher for diabetic than for control rats. In addition, there was no indication of localized dye leakage into retina through defects in the pigment epithelial and vascular endothelial barriers or of increased dye entry at the optic disc, a site of blood-retinal barrier discontinuity. Normalization of tissue fluorescence intensity measurements at the different time intervals to compensate for disparity in concurrent plasma dye concentrations resulted in significantly higher levels in diabetic retinas at 1 hr. However, because this difference was not manifest when the plasma dye concentration vs. time integral was used to normalize the data, it is concluded that no greater accumulation of carboxyfluorescein occurs in the retina of diabetic rats over the time period studied. PMID- 2976363 TI - Multiple small papular seborrheic keratosis. PMID- 2976365 TI - [Intraoperative laser coronary angioplasty. Report of 6 cases]. PMID- 2976364 TI - Evolutionary diversity of reverse (R) fluorescent chromosome bands in vertebrates. AB - Mitotic chromosomes, interphase cell nuclei, and male meiosis of 41 species representing all vertebrate classes were analyzed with distamycin A/mithramycin counterstaining. The purpose of the study was to recognize differences and common characteristics in the reverse (R) fluorescent banding patterns in the chromosomes of vertebrate species at various stages of evolution. In contrast to the warm-blooded mammals and birds, the euchromatic segments in the chromosomes of most reptiles, amphibians, and fishes contain no multiple fluorescent R-bands. This is thought to be due to the absence of the long homogeneous regions (isochores) in the DNA of the cold-blooded vertebrates. Distamycin A/mithramycin banding specifically reveals the GC-rich constitutive heterochromatin in all vertebrates. In most of the vertebrate chromosomes examined, the heterochromatic regions have opposite staining properties with mithramycin and quinacrine. Mithramycin labels the nucleolus organizer regions very brightly in the karyotypes of fishes, amphibians, reptiles and birds, but not of mammals. The lack of mithramycin fluorescence at the nucleolus organizer regions of mammals is attributed to the relatively low level of redundancy of the GC-rich ribosomal DNA in their genomes. Studies on the various meiotic stages of the cold-blooded vertebrates show that the mithramycin labeling of the nucleolus organizers is independent of their state of activity. This can be confirmed by mithramycin fluorescence at the nucleoli of actinomycin-treated cells. PMID- 2976366 TI - [Plasma atrial natriuretic peptide in patients with acute myocardial infarction]. PMID- 2976367 TI - [Studies of hemodynamics with renal vein renin ratio before and after percutaneas transluminal renal angioplasty (PTRA)]. PMID- 2976368 TI - [Advances in research on the mechanism of immunosuppression after traumatic injury]. PMID- 2976370 TI - [Electrocardiographic research on left ventricular hypertrophy in compensated cardiac patients with essential hypertension--comparison between the Fisher's discriminative analysis and echocardiography]. PMID- 2976369 TI - [An immunogenetic study of anterior uveitis]. PMID- 2976371 TI - [Advances in percutaneous balloon catheterization in the treatment of cardiovascular diseases]. PMID- 2976372 TI - Paresthesia and back pain in a patient receiving vancomycin during hemodialysis. AB - A 36-year-old woman was admitted for initiation of hemodialysis for chronic renal failure. Two days after catheter placement the patient developed a fever that persisted and resulted in subsequent removal of the catheter. Although blood cultures were negative, cultures of the catheter tip were positive for Staphylococcus epidermidis. An initial vancomycin dose was well tolerated, but the patient later experienced numbness and tingling of her lower back accompanied by pain ten minutes after initiation of the second dose. Symptoms abated when the vancomycin infusion was discontinued, and the drug was subsequently well tolerated when reinstituted at a slower infusion rate. Similar symptoms were observed five minutes into a vancomycin infusion a week later that also resolved after decreasing the infusion rate. Patients on hemodialysis receiving vancomycin should be carefully monitored during drug administration for the development of paresthesia and spasmodic lower back pain. PMID- 2976373 TI - Diagnostic yield and endoscopic patterns of laparoscopy in the diagnosis of granulomatous hepatitis. AB - In reviewing 54 cases of granulomatous hepatitis (GH) diagnosed among 2,620 laparoscopies, we have been able to classify the capsular GH changes into five patterns: Macular, exudative, pinpoint, granular and cord-like. The exudative pattern was the most frequent one, and predominated in brucellosis. The granular pattern was most common in tuberculosis. Furthermore, 35 additional cases, in whom a laparoscopic diagnosis of GH was not confirmed by liver biopsy, showed a trend to a final diagnosis consistent with multifocal diseases, whenever laparoscopic pictures were similar to the patterns described herein. We speculate with the limits associated with the use of only a single biopsy sample. We conclude that laparoscopy permits the establishment of a tentative diagnosis of GH, which could provide us with valuable information about the usefulness of obtaining multiple hepatic biopsies. PMID- 2976374 TI - Smooth liver surface may conceal cirrhosis. Evidence for the late development of nodular surface of the cirrhotic liver. AB - A 22-year-old male student with HBs Ag + acute hepatic necrosis was admitted to our clinic for further examination. Since his liver function worsened, laparoscopy with direct vision hepatic biopsy was performed. Macroscopically, the surface of the liver did not show the characteristic nodular appearance, in contrast to histological alterations suggestive of liver cirrhosis. Eight months later, a second laparoscopic/bioptic examination was performed. The liver surface appeared nodular, while the histological picture was unchanged vis-a-vis the first examination. The discrepancy between the macroscopic findings observed at the first laparoscopy and those seen eight months later, would suggest that cirrhosis initially develops "inside" the liver and can only be detected histologically; the characteristic nodularity of the liver surface "emerges" subsequently. PMID- 2976375 TI - Reactivation of cell-free models of Physarum plasmodia after myosin reconstitution. AB - Thin-spread glycerol-extracted Physarum plasmodia were treated with N ethylmaleimide (NEM) to block myosin-ATPase and contractility. After supplementing the models with purified plasmodial myosin, they could be reactivated and contracted upon addition of ATP. Fluorescently labeled actomyosin fibers ruptured during contraction, resulting in beaded or rod-like contraction centers. Glycerol-extracted plasmodia lose their negative Ca++-dependence during extraction. Reconstitution of NEM-treated models with plasmodial myosin partly restored this Ca++-sensitivity. Thus, either myosin or a factor associated with it seems to be involved in the Ca++-dependent regulation of cytoplasmic actomyosin contraction in Physarum. NEM-blocked models reconstituted with skeletal muscle myosin were not reactivated by ATP. The same plasmodia subsequently incubated with plasmodial myosin were able to contract. PMID- 2976376 TI - Report of the Joint International Society and Federation of Cardiology/World Health Organization Task Force on Coronary Angioplasty. PMID- 2976377 TI - Inhibition of vasopressin-stimulated flank marking behavior by V1-receptor antagonists. AB - Flank marking, a form of olfactory communication displayed by hamsters, is dependent upon vasopressin-sensitive neurons in the anterior hypothalamus. In the present study two vasopressin type-1 (V1) receptor antagonists, d(CH2)5Tyr(Me)AVP and dPTyr(Me)AVP were tested for their ability to block flank marking stimulated by the microinjection of arginine vasopressin (AVP) into the anterior hypothalamus. Dose-response curves were established for AVP and flank marking in the presence or absence of different concentrations of each antagonist. DPTyr(Me)AVP was microinjected into the anterior hypothalamus 1 h before the microinjection of AVP while d(CH2)5Tyr(Me)AVP and AVP were prepared together and delivered as a single microinjection. This procedure was necessary because dPTyr(Me)AVP, but not d(CH2)5Tyr(Me)AVP, had agonist activity when initially injected into the anterior hypothalamus in concentrations ranging from 0.90-900 microM. The ED50 values (microM) for dPTyr(Me)AVP and AVP were 17.9 and 0.90, respectively. The initial agonist activity of dPTyr(Me)AVP was always followed by blocker activity. Both V1-receptor antagonists caused a dose-dependent decrease in AVP-stimulated flank marking. Maximal inhibition of AVP-stimulated flank marking was produced with approximately 1.0 mM of either antagonist. Both antagonists blocked AVP-stimulated flank marking behavior for over 12 h following their microinjection. PMID- 2976379 TI - [Nondrug therapy of gynecologic diseases]. PMID- 2976378 TI - Effect of streptozotocin on transaminases, creatinine and urea in serum of rats. AB - This study investigates the diabetes-induced lesions in liver and kidney and in addition the possible side effects of the diabetogenic substance streptozotocin (SR) on these organs in non-diabetic animals. 5-week-old female Wistar rats were injected 65 or 130 mg SR/kg body mass. Some animals of the drug group did not become hyperglycemic; thus it was possible to separate the drug effect from the diabetic influence on liver and kidney. In serum investigations some metabolic changes concerning the activities of the liver enzymes aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and the concentrations of urea and creatinine up to 30 days after drug application were studied. SR in hyperglycemic animals causes a time and dose dependent rise in all investigated parameters. Also in normoglycemic rats a significant increase in alkaline phosphatase and in creatinine was observed after 10 days. After 21 and 30 days there were no differences compared to untreated control rats, whereas elevated levels were observed in the hyperglycemic rats. Thus our results support the view of a short damaging effect of SR on liver and kidney without inducing a diabetic state; in hyperglycemic rats the damaging effect is more pronounced. PMID- 2976380 TI - Two cases of large placental chorioangioma with fetal and neonatal complications. AB - Two cases of infants born from pregnancies complicated by large placental chorioangiomas causing congestive heart failure in the neonate as the main complication are presented. Chorioangiomas may be diagnosed early in pregnancy by ultrasound examination. Since both their maternal and their neonatal complications may indicate premature termination of the pregnancy or be conducive to premature birth, repeated ultrasound examinations, including fetal echocardiography, are suggested in order to optimize the timing of the delivery. PMID- 2976381 TI - Alterations in lactate dehydrogenase isoenzymes and phosphofructokinase activity in the hormonally induced G-XII cell line. AB - G-XII cell line was earlier developed in our laboratory from the granulosa lutein cells of goat ovaries, subjected to sustained stimulation by ovine luteinizing hormone. It is a non-clonogenic and non-tumorigenic cell line which secretes progesterone. In the present paper, we report some biochemical findings on the cell line studied between passages 10 and 15. The cells exhibited a shift in the lactate dehydrogenase isoenzyme pattern from the heart form to muscle form and a 300% increase in the phosphofructokinase activity compared to fresh granulosa cells. PMID- 2976382 TI - Prevalence of mental retardation related to fragile X syndrome and other chromosomal abnormalities in the Republic of San Marino. AB - The prevalence of mental retardation related to chromosomal abnormalities, including fragile X syndrome, was studied among 2735 males aged between five and 20 years living in the Republic of San Marino. Five cases of chromosomal abnormalities were found, including one case with fragile X syndrome, with partial epilepsy and a CT scan showing a large arachnoid cyst in the posterior fossa. The prevalence of mental retardation due to chromosomal abnormality was 1.83 per 1000, and that due to fragile X syndrome was 0.37 per 1000. Although these findings relate to only a small population, they are the first from a Mediterranean area. PMID- 2976383 TI - Preventing youth suicide: education is the key. PMID- 2976384 TI - Effect of ionizing radiation on the gastrointestinal tract. PMID- 2976385 TI - Histochemical study of the forelimb muscles in Rana temporaria L. AB - Fibre types in the 7 forelimb muscles of Rana temporaria were characterized according to actomyosin ATP-ase activity and 5 distinct fibre types were distinguished. Besides the twitch fibres of type I, IIA, IIB, and IIC, also the tonic fibres were found in the examined muscles. The metabolic profiles of these fibre types was identified on the basis of SDH activity. The scores of SDH activity of different fibre types overlapped and correlation between fibre types defined by ATP-ase activity and their metabolic profiles was not found. All investigated muscles showed the heterogenous distribution of fibre types. Caput superius m. extensoris carpi radialis and caput mediale m. anconei were composed of 4 and 3 fibre types, respectively, in the opposite to the other ones which contained all 5 types of fibres. PMID- 2976386 TI - Atrial natriuretic hormones. PMID- 2976387 TI - Functional significance of subtypes of 5-HT receptors in the rat spinal reflex pathway. AB - 1. The functional significance of subtypes of 5-hydroxytryptamine (5-HT) receptors was studied in the rat spinal reflex pathway. 2. Ketanserin had no effect on the mono- (MSR) or polysynaptic reflex (PSR) in spinal rats, but decreased the PSR in intact rats. 3. 8-Hydroxy-2-(di-n-propylamino)tetralin (8-OH DPAT) and 5-methoxy-N,N-dimethyltryptamine (5-MeODMT) decreased the MSR and increased the PSR in spinal rats. 4. Ketanserin antagonized the effects of 5 MeODMT without antagonizing the effects of 8-OH-DPAT. 5. Cinanserin had similar effects to those of ketanserin. 6. These results suggest that both 5-HT1A and 5 HT2 receptors mediate MSR inhibition and PSR augmentation in the spinal reflexes of spinal rats, and that the 5-HT2 receptor has a supraspinal tonic excitatory influence on the PSR in intact rats. PMID- 2976388 TI - Effect of (+)cyanidanol-3 on cellular immune reactions and on superoxide dismutase activity in vitro. AB - Immunological and antioxidant effects of the widely used hepatoprotective agent (+)cyanidanol-3 were studied in a complex in vitro test system using isolated peripheral blood lymphocytes of patients with chronic alcoholic liver disease and of healthy donors. In both groups in vitro treatment with (+)cyanidanol-3 inhibited lectin-induced lymphocyte blast transformation and chemiluminescence in a dose-dependent fashion and increased the superoxide dismutase-expression of lymphocytes without influencing the percentage of T and active T cells and antibody-dependent and spontaneous lymphocytotoxicity. The originally lower Con A induced T cell mediated lymphocytotoxicity of patients was completely abolished, while no significant effect was seen using healthy donors' lymphocytes. (+)cyanidanol-3 markedly decreased the Con A-induced suppressor activity of patients' lymphocytes, without influencing that of normal cells. These results suggest that both antioxidant and immunomodulatory effects play an important role in the mechanism of action of (+)cyanidanol-3. PMID- 2976389 TI - Peripheral blood lymphocyte changes after benign gastrointestinal surgery. Role of thymostimulin in reversing the postoperative immunodepression seen in humans. AB - A generalized state of immunosuppression during surgery has been implicated in the development of septic complications postoperatively. We studied 18 patients operated upon for benign diseases, to examine the influence of surgical trauma on circulating lymphocyte subpopulations in man. Additionally, we evaluated the effect of thymostimulin on these changes, in another group of 10 patients. Our results suggest that the total number of lymphocytes, as well as lymphocyte subpopulations CD3+ and CD4+, fell significantly following surgery. This reduction in cell number is more pronounced on the helper/inducer lymphocytes. The CD4+/CD8+ ratio decreases significantly after operation. For patients with no complications, the immunosuppression in terms of peripheral lymphocyte population, seen in the postoperative period is usually reversible around the seventh day. On the other hand, peripheral blood lymphocyte changes in the postoperative period were less pronounced in patients treated with thymostimulin. Thymostimulin's action is predominantly on T lymphocytes, and within these, on the T helper/-inducer subpopulation. And finally, thymostimulin is capable of maintaining a normal relationship between helper/inducer and suppressor/cytotoxic cells (CD4+/CD8+ ratio) during the postoperative period, suggesting a better immune state. PMID- 2976390 TI - Percutaneous transluminal coronary angioplasty (PTCA)--application in evolving myocardial infarction. PMID- 2976391 TI - Early experience on percutaneous transluminal coronary angioplasty. PMID- 2976392 TI - [Laparosplenectomy in Hodgkin's disease]. PMID- 2976393 TI - Functional plasticity of microglia: a review. AB - The present review summarizes recently acquired data in vivo, which support a role of CNS microglia as a source of defense cells in the CNS capable of carrying out certain immune functions autonomously. We have kept the following discussion restricted to microglial cells and have not included work on the immunological functions of astrocytes, which has been recently reviewed elsewhere (Fontana et al.: Immunological Reviews 137:3521-3527, 1987). Resting microglia are scattered uniformly throughout the CNS forming a network of potential immunoeffector cells, which can be activated by stimuli ranging from peripheral nerve injury over viral infections to direct mechanical brain trauma. The term "activated microglia" is used here to describe proliferating cells that demonstrate changes in their immunophenotype but have not undergone transformation into brain macrophages. Such a transformation can be stimulated by neuronal death but not by sublethal neuronal injury. Microglia may function as antigen-presenting cells and may thus represent the effector cell responsible for the recruitment of lymphocytes to the brain resulting in an inflammatory reaction. The recent developments in the understanding of microglial cell function may lead to a redefinition of the often cited "immune privilege" of the brain. PMID- 2976394 TI - Prenatal gliogenesis in the developing cerebrum of the mouse. AB - A correlative light microscopic, ultrastructural, and immunocytochemical study was made of the brains of C57BL/6J mice obtained between embryonic day (E-) 11 to postnatal day (P-) 3. The deployment of radially oriented glial cells within the neopallium was already apparent by E-12, at which time the expanded endfeet of processes abutting the basement membrane at the pial surface showed ultrastructural evidence of glial differentiation. Scattered, horizontally arranged glial cells were also observed within the marginal zone prior to the arrival of the cortical plate neurons. Indirect immunofluorescence for glial fibrillary acidic protein (GFAP) was detected at the outer margins of the neopallium at E-12, antedating the expression of GFAP in radially organized fibers. Radial fibers traversing the full thickness of the neopallium became strongly immunoreactive for GFAP only at and after day E-16. These findings provide evidence of early gliogenesis in the developing mouse forebrain and indicate that GFAP is as readily demonstrable in mouse astrocytes and radial glial fibers as it is in those of man and subhuman primates. PMID- 2976395 TI - Effect of thyroid deficiency on the development of glia in the hippocampal formation of the rat: an immunocytochemical study. AB - The development of glia in the hippocampal formation of normal and hypothyroid rats was studied using immunocytochemical staining for either glial fibrillary acidic protein (GFAP) or vimentin. Light microscopy showed lower GFAP immunoreactivity in the radial glial processes of young hypothyroid rats compared to normal animals. These processes followed the known path of neuroblast migration toward the proliferative zone of the dentate gyrus until the end of the 1st postnatal week. Vimentin immunoreactivity showed that the glial processes were present and therefore immature at least with respect to their cytoskeletal composition. We propose that this early defect in the maturation of the radial glial fibers accounts for the final deficit in the granule cells of the dentate gyrus. Later in development, thyroid deficiency also reduced the density and number of GFAP-labeled astrocytes and the growth of their processes. This observation is in complete disagreement with the glial hypertrophy induced by thyroid deficiency in the cerebellum. The considerably increased histogenetic cell death observed in the cerebellum of young hypothyroid rats could in turn induce glial hypertrophy, whereas the hippocampal formation, where a normal low number of cell deaths is observed, is only subjected to the general depressive effect of thyroid deficiency on cell maturation. PMID- 2976396 TI - Lactate release from cultured astrocytes and neurons: a comparison. AB - Lactate released into the surrounding salt solution as well as the cellular lactate content were measured in cerebral primary cultures of mouse astrocytes and of mouse neurons. Any newly produced lactate was immediately released as lactic acid into the extracellular compartment via a lactate/proton cotransport. The astrocytic release was about 2,000 nmol x mg-1 x hr-1; the neuronal release was about 300 nmol x mg-1 x hr-1. However, if election transport was blocked with dinitrophenol, the neuronal lactate release was as high as the astrocytic one under normal conditions. High glucose (30 mM) and K+ (60 mM) increased lactate release of astrocytes but not of neurons. In contrast it was found that insulin (1 microM) exposure mainly stimulated neuronal lactate release rather than glial release. Adenosine stimulated both neuronal and glial release. Neither intracellular lactate content nor concentration changed significantly in either cell type under any conditions tested. The pathophysiological implications of these measurements are discussed. PMID- 2976397 TI - Lactic acid inhibition of gap junctional intercellular communication in in vitro astrocytes as measured by fluorescence recovery after laser photobleaching. AB - Lactic acid can permeate plasma membranes, causing intracellular acidosis. Gap junctions are sensitive to pHi and can be reversibly uncoupled by weak acids. In this study, dye coupling between in vitro astrocytes, presumably mediated by gap junctions, was measured in the absence and presence of lactic acid. Fluorescence recovery after laser photobleaching (gap-FRAP analysis) was used to measure dye coupling. Astrocytes bathed in Eagle's minimum essential medium (EMEM) with lactic acid, pHo 5.5-6, showed no difference in their dye coupling (mean recovery of fluorescence 30%) when compared to control astrocytes (mean recovery of fluorescence 26%). However, 24 mM lactic acid in EMEM, pHo 4.5, decreased dye coupling (mean recovery of fluorescence 2.0%). This effect occurred within 5 min of treatment. When lactic acid-EMEM, pH 4.5, was removed from astrocytes after 30 min and the cells were incubated in EMEM for 24 hr, decreased coupling was not reversed (mean recovery 4.0%). When lactic acid-treated astrocytes were incubated in EMEM for 48 hr, the mean recovery of fluorescence increased to 15% (i.e., 42% of the recovery seen in controls). These observations suggest that brief exposure to high concentrations of lactic acid can have immediate and long-lasting effects on glial gap junctional communication. Under pathological circumstances, such a sequence could be initiated, and this might impair astrocytic control of the central nervous system microenvironment mediated by spatial buffering. PMID- 2976398 TI - Glial cells promote dendritic development in rat sympathetic neurons in vitro. AB - Many types of glial-neuronal interactions occur during the development of the nervous system. To determine how such interactions might affect the development of autonomic ganglia, we compared the morphology of embryonic rat sympathetic neurons grown in the absence and in the presence of ganglionic nonneuronal cells in serum-free medium. Dye injections, electron microscopy, and immunocytochemistry were used to distinguish axons from dendrites. In cultures without nonneuronal cells, most (greater than 80%) sympathetic neurons extended only a single axonal process, and this unipolar state persisted for at least 8 weeks. Coculture with ganglionic nonneuronal cells caused sympathetic neurons to become multipolar and to extend multiple (range 1-17) dendrites. Morphometric measurements made after 1 month of coculture indicated that the amount of dendritic growth that occurred in vitro (mean number of dendrites/cell = 7.5; total dendritic length = 1,050 micron) was similar to that normally occurring during a comparable period in situ. In contrast to its prominent effects on dendritic growth, coculture did not cause changes in the number of axons/neuron or in the uptake of neurotransmitter. Cultures with ganglionic nonneuronal cells were immunostained for antigens present on the surfaces of fibroblasts (Thy-1.1, fibronectin) and of glia of the peripheral nervous system (laminin). Fewer than 1% of the nonneuronal cells displayed immunoreactivity for fibroblastic antigens; in contrast, greater than or equal to 99% reacted with antibody to laminin. Moreover, reconstitution experiments revealed that purified populations of laminin-positive Schwann cells promoted dendritic growth. Fibroblasts and heart cells lacked this activity. These data indicate that glia selectively promote dendritic development in sympathetic neurons maintained in serum-free medium. PMID- 2976399 TI - Glial fiber pattern in the developing chicken cerebellum: vimentin and glial fibrillary acidic protein (GFAP) immunostaining. AB - The possible relation between glial fibers and the formation of longitudinal granule cell migration patterns that occur in the cerebellar anlage of the chicken was investigated by immunocytochemistry of vimentin (monoclonal antibody) and glial fibrillary acidic protein (polyclonal antibody against GFAP, PGF) on fixed and unfixed brain tissues. In addition, neuronal development was studied with a monoclonal antibody for neurofilament. Vimentin was present in radial and tangential fibers in the cerebellar anlage during granule cell migration in almost all parts of the anlage. However, no specific topographic relation of vimentin and GFAP to the migration pattern of granule cells was observed. In adults, Bergmann fibers and astroglia were stained with vimentin antiserum and not with GFAP antiserum. Conclusions are that radial fibers do not determine the formation of longitudinal cytoarchitectonic patterns in the chick cerebellum and that vimentin is the main cytoskeletal component of Bergmann fibers and astroglial cells in embryonic and adult chicken cerebellum. PMID- 2976400 TI - Pure astrocyte cultures derived from cells isolated from mature brain. AB - Enriched preparations of oligodendrocytes, isolated either from adult bovine brain or from 30-day-old rat brain, eventually yield cultures in MEM-15% calf serum that contain, in addition to oligodendrocytes, proliferating astrocytes and variable numbers of fibroblast-like cells. If these cultures are switched to a serum-free defined medium during the 1st week, mixed cultures containing only oligodendrocytes and astrocytes are obtained. Bovine cultures can be replated and purified by selective adhesion to yield cultures that are greater than 99% astrocytes; similar procedures were not successful with rat cultures. Cytoskeletal preparations of the purified astrocyte cultures from mature bovine brain contain both vimentin and glial fibrillary acidic protein (GFAP), but vimentin is by far the major intermediate filament protein. Thus, the intermediate filament composition of these astrocytes is similar to that of astrocytes in primary cultures obtained from neonatal rat brain. Immunofluorescent studies of these cultures at 24 hr in vitro show that there are no GFAP+ cells in cultures of either species; the bovine cultures contain greater than 95% GC+ cells; and the rat cultures contain 90% GC+ cells. After a few days in vitro flat cells appear that are vimentin+/GFAP-/GC-. In serum-free medium these cells eventually become vimentin+/GFAP+. We propose that the astrocytes that grow in these cultures arise from a population of glial precursor cells, which are present even in adult brain and are isolated together with oligodendroglia, and that they do not derive from contaminating mature astrocytes. Thus, the astrocytes in our cultures may have the same origin as astrocytes grown in culture from dissociated neonatal brain. PMID- 2976401 TI - Heterogeneity of potassium currents in cultured oligodendrocytes. AB - In the present study we have analyzed the membrane currents of mature oligodendrocytes in cultures from dissociated fetal mouse cerebral hemispheres and explant cultures from fetal mouse spinal cord. Both types of oligodendrocytes showed large voltage-dependent, but time-independent inward and outward currents that were partially blocked by Ba2+. In addition, time- and voltage-dependent inward and outward currents were observed in a minority of oligodendrocytes from spinal cord. All voltage-dependent currents were completely blocked by Ba2+, and inward currents were completely blocked by Cs+, suggesting that they are mediated by K+ channels. Current-voltage curves of mouse spinal cord oligodendrocytes varied from being linear to outwardly or inwardly rectifying. In contrast, oligodendrocytes cultured from mouse brain always showed an inward rectification of the current voltage relation and a lack of time-dependent currents. It thus appears that mature oligodendrocytes in explant cultures of mouse spinal cord, in contrast to oligodendrocytes from dissociated brain, consist of different cell populations that are distinguished by their expression or active state of K+ channels. PMID- 2976402 TI - Induction of T-helper cell activity by fragments of rye grass pollen extract produced by digestion with chymotrypsin. AB - Rye grass pollen extract was digested by chymotrypsin to produce fragments with a molecular weight below 10,000, as demonstrated by polyacrylamide gel electrophoresis. Chymotryptic fragments did not react with either human or mouse IgG antibodies specific for rye grass pollen, nor did they induce an antibody response in mice with specificity for the parent extract. However, macrophage presented fragments retained the ability to react with rye-specific T cells in a lymphoproliferation assay. Furthermore, these fragments induced the development of splenocytes capable of supporting dinitrophenyl specific antibody production. This implies that the fragments still react with, and induce, rye grass pollen extract-specific T-helper cells. The possibility that such fragments might have potential for use in immunotherapy for the specific treatment of allergy is discussed. PMID- 2976403 TI - Experimental ventricular hypertrophy in growing pigs. AB - An experimental model has been developed in the growing pig to study the pathology of myocardial hypertrophy. Aortopulmonary shunts were created in 4-week old piglets and the animals were followed-up to 7 months of age. The pulmonary arterial pressure had increased at 4 weeks of shunt function by an average of 118% and the pulmonary blood flow by as much as 63%. In the longest living animal (7 months), the pulmonary arterial pressure and blood flow had increased by 170 and 53%, respectively, in relation to normal findings. The animals were killed at varying intervals and the hearts removed and examined. The weight of the entire hearts and their individual component parts increased significantly when compared with values found for normal postnatal development (P less than 0.001). The percentage increase in the total heart weights increased at all stages of development varying from an initial 109% to 39% at the time of killing. All the other parameters were also significantly greater than the values found for the normals, including the myocardial wall thickness in both ventricles (P less than 0.001). Histology showed changes comparable to gross myocardial wall thickness hypertrophy observed in human congenital anomalities. The method is recommended as a model for further studies on myocardial hypertrophy in growing individuals. PMID- 2976404 TI - Pulmonary valvoplasty--experience of 100 cases. AB - One-hundred patients with pulmonary valve stenosis underwent pulmonary valvoplasty, their ages ranging from 1 to 59 years. The systolic gradient across the valve ranged from 47 to 260 mm Hg (97.67 +/- 41.15) prior to the valvoplasty, and from 0 to 55 mm Hg (14.72 +/- 11.40) immediately after dilatation (P less than 0.0001). The clinical follow-up of 18.2 months of 56 patients showed a tendency for the systolic thrill to disappear, the systolic murmur became softer and there was a tendency to normalization of the electrocardiogram. A hemodynamic restudy was carried out in 54 patients and the systolic gradients across the valve ranged from 21.55 +/- 23.86. Seven patients required redilatation. In patients with hypertrophy of the infundibulum prior to valvoplasty this was observed to regress, indicating that infundibular hypertrophy is reversible. PMID- 2976405 TI - Scleredema adultorum of Buschke and primary hyperparathyroidism. PMID- 2976406 TI - Acitretin (Ro 10-1670) in the treatment of severe psoriasis. A randomized double blind parallel study comparing acitretin and etretinate. AB - A randomized double-blind parallel trial comparing acitretin and etretinate was performed in 20 patients with severe psoriasis during a treatment period of 12 weeks. The initial dose was 30 mg/day for 4 weeks, and the mean dose at the end of the study was slightly lower in the acitretin group when compared to the etretinate group (30.7 mg/day and 33.4 mg/day, respectively). Follow-up examinations were carried out every 2 weeks, and the efficacy of treatment was evaluated by using the PASI score. Percentage improvement in the PASI score was 50% at week 10 in both groups, and the difference between them at week 12 was insignificant. Both quantitatively and qualitatively, acitretin and etretinate do not significantly differ. PMID- 2976407 TI - Acne therapy (1934-1949). A review of 50 records. PMID- 2976408 TI - Toxicity of intravitreal injection of foscarnet in the rabbit eye. AB - Foscarnet (sodium phosphonoformate) is an antiviral with a high degree of efficacy against members of the herpes simplex virus family. We studied the toxicity of single doses of intravitreally administered foscarnet in the albino rabbit. Eyes were evaluated clinically and by light microscopy. Data demonstrated that doses ranging from 20 to 1000 micrograms per 0.1 milliliter are nontoxic to the retina, suggesting that foscarnet may be useful in the treatment of acute retinal necrosis and cytomegalovirus retinitis in patients with acquired immunodeficiency syndrome. PMID- 2976409 TI - Microtubules in cardiac myocytes. PMID- 2976410 TI - Trimetrexate in advanced carcinoma of the esophagus. AB - Twenty-four patients with advanced epidermoid carcinoma of the esophagus were treated with trimetrexate (TMTX), a lipid soluble non-classical antifol. Patients were given TMTX 8 mg/m2 intravenously day 1-5 every 28 days. In nine of these patients the dose was escalated to 12 mg/m2 day 1-5 every 28 days. Three patients had a partial response (95% confidence limit 3-32%) with a median response duration of 14 weeks. No hematologic toxicity was documented. Two patients developed moderate stomatitis and only 3 patients experienced any nausea or vomiting. The median survival of all patients is 12 weeks. It is concluded that a higher dose of TMTX should be studied in patients with esophageal cancer in order to assess the true therapeutic value of the agent at a dose closer to the median tolerated dose. A phase II ECOG study using TMTX 12 mg/m2 intravenously day 1-5 every 21 days is currently being conducted. PMID- 2976411 TI - Cardiac disease associated with the acquired immunodeficiency syndrome: a case report and review of the literature. PMID- 2976412 TI - Protein gene product 9.5 (PGP 9.5). A new neuronal marker visualizing the whole uterine innervation and pregnancy-induced and developmental changes in the guinea pig. AB - The guinea pig uterus is supplied by different populations of nerves which can be demonstrated by specific immunocytochemical and histochemical techniques. So far, there has been no single marker displaying entire peripheral innervation patterns. Recently, protein gene product (PGP) 9.5, a cytoplasmic protein in neurons and neuroendocrine cells, was found to visualize both different populations and subtypes of nerves. This prompted the present study of using PGP 9.5 for visualization of the whole uterine innervation. This was performed by the indirect immunofluorescence method using antiserum to PGP 9.5 raised in rabbits. PGP-immunoreactivity was present in all neuronal parts of the extrinsic and intrinsic uterine innervation, including different subpopulations of nerves. This was verified by chemical sympathectomy and sensory denervation with 6 hydroxydopamine and capsaicin-treatment respectively, and double immunostaining. By term a disappearance of uterine PGP-nerve-immunoreactivity was observed which was almost complete in fetus-bearing uterine tissue and further strengthens previous assumptions of a general, pregnancy-induced uterine neuronal degeneration. The developmental time-course and morphology of PGP-immunoreactive nerve structures was similar to that for other neuronal markers and support the suggestion of PGP-immunoreactivity as a general marker for the entire uterine innervation, and suggests that the presence of PGP 9.5-immunoreactivity may coincide with functional maturation of uterine innervation. PMID- 2976413 TI - Human suppressor T cells induced in vitro with an autologous renal allograft derived T cell line. I. Suppressor cell induction, function, and specificity. AB - The functional characteristics of T suppressor (Ts) cells generated from the peripheral blood lymphocytes (PBL) of a kidney transplant recipient who had excellent graft function for 1 year were examined. Ts cells were induced by co culture of PBL with an autologous alloreactive cytotoxic T lymphocyte (CTL) line (EE-1) previously grown from a routine renal allograft biopsy of this patient performed 10 days posttransplant. The EE-1 line included CD3+ T cells of CD8+ and CD4+ phenotypes with cytotoxic specificity for disparate class 1 (HLA-B8) and class II (HLA-DR1 and 3) antigens of the kidney donor (JC). The EE-1 induced Ts cell lines (designated TsEE) were found to significantly suppress (50%-95%) autologous fresh responder EE-PBL stimulation by donor EBV-transformed cells (JC EBV) in mixed lymphocyte reaction (MLR) assay. TsEE cells were CD3+ (98%) and predominantly CD8+ (68-80%), showed no cytotoxic activity, and were suppressive only at the early phase of MLR stimulation. In three-party cell test MLR assays, TsEE-mediated suppression appeared restricted to responder cells sharing HLA-B7 with the suppressor line, and was not abrogated by the addition of exogenous interleukin-2 (IL-2). TsEE cells also showed restricted suppression of CTL generation but not mature CTL activity. The restricted suppressor activity of TsEE lines was dependent upon their induction and restimulation with the autologous EE-1 line. PMID- 2976414 TI - Combined GnRH-agonist/gonadotrophin stimulation for in-vitro fertilization. AB - The occurrence of a premature luteinizing hormone (LH) surge during gonadotrophin stimulation for in-vitro fertilization leads to cancellation of the cycle. Moreover, insufficient follicular maturation is often caused by elevated basal gonadotrophin levels. Therefore, the gonadotrophin-releasing hormone (GnRH) agonist, D-Trp-6-LHRH, was applied to patients exhibiting premature LH surges, hyperandrogenaemia or incipient premature menopause. A total of 119 cycles were treated using a long-acting versus a short-acting GnRH agonistic analogue. In protocol 1, patients received daily s.c. injections of 100-500 micrograms of a short-acting compound. In protocol 2, a long-acting bolus of 3.2 mg was given i.m. Concomitant human gonadotrophin stimulation was started in protocol 1 after clinical and biochemical evidence of pituitary suppression and in protocol 2 after a fixed suppression interval of 14 days. In protocol 1, higher oestrogen levels were reached with more oocytes harvested. The pregnancy rate per transfer was increased from 3.5 to 18%, with most pregnancies occurring with protocol 1. The cancellation rate of 13.4% was mainly due to insufficient follicular development in patients in whom premature menopause was suspected. Hyperandrogenaemic patients with an elevated LH/FSH ratio exhibited the best follicular recruitment with the highest pregnancy rate of 25% per transfer. Thus, combined GnRH-agonist/gonadotrophin stimulation offers a causal treatment for patients susceptible to premature LH surges and for hyperandrogenaemic patients. PMID- 2976416 TI - [The place of immunology in recurring spontaneous abortion]. AB - Spontaneous repeated miscarriages are often explained by an immunological mechanism. Whereas in normal pregnancy the mother develops a tolerance immune response induced by paternal antigens of fetus, she is unable to react in this variety of miscarriage. The immunological theory is supported by some solid experimental arguments, which are detailed. Antigens generating the tolerance response are probably TLX antigens, expressed on syncitiotrophoblast and cross reacting with class I HLA antigens. Diagnosis of immunological miscarriage is based on elimination of other causes and on absence in woman of antibodies directed against husband class I antigens. Tolerance immune response can be induced by injections to wife of great quantity of conjunct lymphocytes. An anti HLA, and later on anti TLX, immunization is often obtained. Several teams have applied this treatment, with frequent good results: about 80% of fecundated patients conduct a normal pregnancy until its time. PMID- 2976415 TI - Inadvertent intrathecal administration of amidetrizoate. AB - Two cases are presented in which amidetrizoate (Urografin) was accidentally introduced into the intrathecal cavity. Intrathecal lavage and continuous administration of thiopentone were very successful in preventing further systemic deterioration. PMID- 2976417 TI - [Comparison of the results of umbilical artery velocimetry obtained by pulsed Doppler and continuous Doppler]. AB - Doppler assessment has become the fashion in obstetric screening at present and this is absolutely justified in view of the quality of the information that is obtained. The authors have tried to assess the two methods of analysing the blood flow through the umbilical artery by using pulsed Doppler and continuous Doppler techniques. They have used a fairly new statistical method (differential). The authors have found that the methods are interchangeable. They did this on 32 patients who on the same day had pulsed and continuous Doppler tests and they used Pourcelot's placental resistance technique. These observations show that consistent diagrams which are equivalent to one another are obtained by the two methods and they can be used economically. PMID- 2976418 TI - Pelvic examination versus fiberoptic laparoscopy. A fictional study of patient preference in 1,534 women. AB - A fictional study of patient preference was conducted in 1,534 women who received both laparoscopy and a pelvic examination plus pelvic ultrasonography. The overwhelming majority of women (87%) preferred laparoscopy, with most patients (78%) indicating they would undergo a repeat examination if requested. Hospital occupancy rose 24% while gynecologists' incomes rose 58%. Radiologists' incomes declined 14%. Seven deaths occurring in conjunction with laparoscopy were determined to be statistically insignificant. Based on these findings, it is suggested that the pelvic examination be replaced by routine laparoscopy. PMID- 2976419 TI - [Immunohistochemical study of cancer tissues. 1. Special reference to Leu-11b positive lymphocytes (NK cells)]. PMID- 2976420 TI - [Tumor-specific immunotherapy model utilizing virus-reactive helper T-cell activities]. PMID- 2976421 TI - [Iatrogenic Cushing's syndrome with mediastinal lipomatosis simulating cardiomegaly and pleural effusion: report of a case]. PMID- 2976422 TI - Compensating bladder cancer victims employed in aluminum reduction plants. AB - A criterion for eligibility to compensation is sought for bladder cancer cases among workers in the aluminum smelting industry. Probability that a case of bladder cancer was caused by occupational exposure can be estimated from a relationship derived from results of epidemiologic studies. Because the effects of occupational exposure and smoking apparently combine multiplicatively, this probability is independent of whether a case patient smoked. Estimated probabilities of causation have been used in a criterion for eligibility to compensation by the Quebec workers' compensation board. Workers with cancer for whom the upper 95% confidence limit of the probability of causation is at least 50% are compensated. This implies a minimum cumulative exposure to benzo[a]pyrene (concentration in micrograms per cubic meter times duration in years) of 19 micrograms/m3 years. Possible alternative approaches to compensation are discussed. PMID- 2976423 TI - Nurses' beliefs about cause and prevention of occupational back pain. AB - Nurses' beliefs and attitudes about causes and prevention of occupational back pain of nurses (OBPN) were studied by interviewing 68 nurses using a semi structured format. Direct patient contact activities, particularly patient transfers, were most frequently mentioned as causes of OBPN. Attitudes and beliefs about preventive methods focused upon personal work practices and personal behavior such as choice of clothing and personal exercise. The findings of this study suggest that the causes of and preventive methods for OBPN identified by nurses differ from the actual work tasks performed and possibly associated with OBPN as determined by other studies. The authors postulate that nurse training and attitudes lead to a narrow focus; avoiding preconceived concepts is necessary for preventing OBPN. PMID- 2976424 TI - Duration of chronic pain and the Minnesota Multiphasic Personality Inventory: profiles of industrially injured workers. AB - The present study explored the usefulness of the Minnesota Multiphasic Personality Inventory (MMPI) in understanding the relationship between duration of chronic pain and psychiatric difficulties. The MMPI responses of workers' compensation patients with varying levels of low back pain chronicity were compared. One hundred ninety eight patients, divided into three groups according to length of disability, underwent social history interviews and completed the MMPI. In regard to the overall profile, the sample had elevations on the Hypochondriasis, Depression, and Hysteria scales. A series of analysis of variance designs revealed that those who were disabled for two or more years evidenced significantly more depression and psychopathology than those who were disabled for less than one year. Further statistical evaluation of 200 social history variables did not reveal significant differences between the three groups on most variables. In conjunction with previous research, the results suggested a causative link between disability and psychiatric disease. PMID- 2976426 TI - The immune regulation in familial Mediterranean fever (FMF). AB - In order to investigate a possible immune regulation imbalance in familial Mediterranean fever (FMF), the T-cell subsets and interleukin (IL)-1 and -2 production were examined in 39 patients (32 consecutive; 7 previous) and 14 controls. Results in the FMF group indicated no change in total T-cells and B cells. The number of supp T-cells and helper cells were significantly decreased, as compared to the controls (14 +/- 5.2, 19 +/- 4.6 vs. 31 +/- 4.6, 41 +/- 5.3, respectively), and the NK cells were significantly increased (16 +/- 4.8, 36 +/- 2.1). Peripheral blood monocytes from the patients with FMF produced higher amounts of IL-1 and lower amounts of IL-2 than those from the control subjects. The latter results were enhanced when the FMF group was subdivided on the basis of pretreatment with colchicine and presence of amyloidosis. This study, although preliminary, indicates an immune regulation imbalance in FMF patients. Further research is necessary to understand the interrelation of amyloidosis and colchicine treatment. PMID- 2976427 TI - The autologous mixed lymphocyte response: isolation of mononuclear cell populations by counterflow centrifugal elutriation. AB - We have used counterflow centrifugal elutriation (CCE) to isolate cell populations for an autologous mixed lymphocyte response (AMLR). Enriched T lymphocyte and monocyte populations were obtained rapidly without the need for cells to first form rosettes with sheep RBC (T cells) or adhere to plastic (monocytes). In 16 separate experiments, peripheral blood (PB) was separated by CCE (9) and/or a plastic adherence and nylon wool column method (9). Viability by all methods was greater than 95% CCE yielded greater than 97% lymphocytes in F20 and greater than 90% lymphocytes in F26. Cells obtained after passage through nylon wool yielded greater than 90% lymphocytes. Both CCE (F34) and plastic adherence techniques yielded greater than 75% monocytes. Similar AMLR response was obtained using either cell separation method. We conclude that mononuclear cells isolated by CCE are responsive in the AMLR and yield comparable data to other more tedious techniques. PMID- 2976425 TI - Effects of a specific thromboxane synthetase inhibitor on development of experimental Dirofilaria immitis immune complex glomerulonephritis in the dog. AB - Twelve Beagle dogs were immunized with aqueous-soluble Dirofilaria immitis antigens, and subsequent to at least fivefold increases in serum antibody titer, 6 mg of homologous antigen was infused into the left renal artery. Six dogs were treated once daily starting the day of infusion with 0.75 mg/kg of 1 benzylimidazole (1-BIM) in saline. Six control dogs were given saline only. Light, immunofluorescent, and transmission electron microscopic examinations of renal tissue from control dogs, 10 days after antigen infusion, showed a mesangioproliferative glomerulonephritis in the left kidney with polymorphonuclear leukocyte (PMNL) infiltration and fibrin deposition. Immunoglobulin (Ig) G, M, C3, and Dirofilaria antigen deposits were observed in a segmental granular pattern. Mesangial, subendothelial, and intramembranous electron dense deposits were observed, and anti-Dirofilaria antibodies were demonstrated in kidney eluates from each dog. Administration of 1-BIM had no significant effect on IgG, IgM, C3, or antigen deposits, electron dense deposits, or concentration of antibody in kidney eluates. However, 1-BIM-treated dogs had less glomerular cell proliferation, periodic acid-Schiff (PAS) positive glomerular staining, PMNL infiltration, and fibrin deposition. These data suggest that thromboxane is an important mediator in the development of immune complex glomerulonephritis, and that in certain circumstances, inhibition of thromboxane synthesis may be an effective therapy for immune complex glomerulonephritis in the dog. PMID- 2976428 TI - The changes in plasma lipoproteins in rat with diabetes induced by streptozotocin. PMID- 2976429 TI - Limitations of ABR as a hearing test as exemplified in multiply handicapped adults. AB - Some limitations of the audiometric application of the click-evoked ABR are exemplified in the results for this sample of multiply handicapped hearing impaired adults. Normal ABR thresholds were not necessarily indicative of normal hearing. Complete absence of ABR was not necessarily associated with total deafness. These findings underscore the need for conservative interpretation when the ABR is used to assess hearing of neurologically impaired patients at risk for more central auditory dysfunction. Additional diagnostic information in problematic cases may be obtained with cortical auditory potentials. Electrophysiological data must, in turn, be integrated with behaviorally obtained results. PMID- 2976430 TI - Surgical management of obstructive sleep apnea in children with Down syndrome. AB - Obstructive sleep apnea (OSA) is relatively common in children with Down Syndrome due to anatomical and immunological variations which place them at particular risk. Secondary cardio-respiratory complications of upper airway obstruction may be found at the time of initial diagnosis; these require medical therapy before surgical correction of the OSA is attempted. The selection of appropriate surgical therapy is influenced by the anatomical structure of the individual child. Tonsillectomy and adenoidectomy alone infrequently provide lasting relief of obstruction for these children. We have found that uvulopalatopharyngoplasty (UPPP), with or without simultaneous anterior reduction of the tongue, alleviates this form of airway obstruction. This paper describes the procedures and reports our experience of six patients with Down syndrome. PMID- 2976431 TI - [Effects of hematoporphyrin derivative (HpD) and laser photoradiation on cultured cells]. PMID- 2976432 TI - [Plasma platelet-derived factors in patients with systemic scleroderma- alterations of plasma beta-thromboglobulin and platelet factor 4 levels after vibration stress on bilateral hands]. PMID- 2976433 TI - [The mechanism of polyuria associated with paroxysmal supraventricular tachycardia]. PMID- 2976434 TI - [Profuse intra-abdominal hemorrhage in laparocentesis of ascites]. PMID- 2976435 TI - [Cholecystostomy through the round ligament of the liver]. PMID- 2976437 TI - Differences in the gastrointestinal microbiota of specific pathogen free mice: an often unknown variable in biomedical research. AB - Large differences were found in the numbers of facultatively anaerobic Gram negative bacteria in the gastrointestinal tract of mice from 3 major specific pathogen free (SPF) units in Australia. The species isolated also differed between mouse colonies. In one unit the presence of Enterobacter cloacae was found to dramatically influence the survival of mice following total body irradiation. This finding conforms with previous studies which have shown the influence of variation in gastrointestinal microbiota on the immune system and on susceptibility to infection. Given that the presence or absence of Enterobacteriaceae in the intestines of mice under investigation may influence experimental results, researchers using SPF rodents are encouraged to determine the baseline loading of these bacteria in their animals. Where results of immunological or irradiation studies from different colonies are likely to be compared, the enterobacterial status of the colony being used should be reported. PMID- 2976438 TI - Perceptual categorization and consistency of synthesized (r-w) continua by adults, normal children and (r)-misarticulating children. AB - The purpose of this study was to determine if children who misarticulate (r) differ from normal children and adults in the perception of sound features that are produced correctly and incorrectly. Children with normal articulation, children who produced (r) misarticulations, and adults listened to synthesized child and adult (r-w) continua in two separate sessions, and to an adult (b-w) control continuum in one session. Perception was evaluated on the basis of measures of phonetic boundary location and the consistency of response to each stimulus in a continuum. The (r)-misarticulating children were found to be significantly less consistent than child and adult controls in responding to the (r-w) stimuli. Moreover, consistency scores were significantly higher for the adult continuum than for the child continuum. The performance of children was different from that of adults. Due to inconsistent performance, boundaries could not be computed for (r)-misarticulating children, but it was found that the boundaries for children in the control group were closer to the (r)-end of the continuum than those for adults. In the case of the (b-w) continuum, it was found that (r)-misarticulating children were significantly less consistent than adults. The phonetic boundaries of children were significantly closer to the (b)-end of the continuum than the boundary for adults. Thus, the results reveal that variability in stimulus response was influenced primarily by the productive ability of the subjects, whereas differences in stimulus categorization were influenced by the age of the subjects. The perceptual variability was most clearly reflected by responses to stimuli produced incorrectly, whereas categorization differences extended to sounds produced correctly. PMID- 2976439 TI - Comment on Cavallo and Baken (1985) PMID- 2976440 TI - [Inhibition of the function of the neutrophil granulocyte caused by autologous serum from patients with acne vulgaris]. AB - Sixteen patients with acne vulgaris were studied. All patients had severe inflammatory lesions, nodulocystic and conglobata. Of them, 12 was male and 4 female, their age was between 16 and 24 years old. In all the cases laboratory test to evaluate chemotactic, nitroblue-tetrazolium and lysosomal activity of granulocyte were performed. The protocol included in vitro incubations with and without autologous serum for considered their action on the phagocytic cell. The results shows an inhibitory action of the autologous serum in chemotactic (d mean, 34.25%; SEM, 2.39, p less than 0.01), nitroblue-tetrazolium test (d-mean, 21.06%; SEM, 2.85, p less than 0.05), and lysosomal activity (d-mean, 21.43%; SEM, 2.80, p less than 0.05). The authors suggest that granulocyte alterations that occur in inflammatory forms of acne vulgaris is a secondary phenomenon may be caused by seric IgE or vasoactive substances released during immune responses. PMID- 2976436 TI - [A method of laparoscopic cholecystostomy]. PMID- 2976441 TI - [The function of the neutrophil granulocyte in acne vulgaris]. AB - Twenty seven patients with acne vulgaris were studied. Of them, 10 had minimal lesions and 17 had severe inflammatory lesions: nodulocystic, and conglobata. NBT and lysosomal neutrophil function was performed. The results shows a significative decreased values for the three test in patients with inflammatory lesions compared to patients with non-inflammatory lesions. The values for chemotactic function was 56.23 and 93%, for NBT test 50.82 and 93% and for lysosomal activity 60.88 and 88.9%, respectively. The authors proposed that the altered in vitro neutrophil function is a secondary phenomenon caused by specific and non-specific immunologic factors that occur in patients with severe inflammatory acne vulgaris. PMID- 2976442 TI - Excimer laser angioplasty: pygmalion makes it to the ball. PMID- 2976443 TI - Molecular, cellular, and tissue responses following photodynamic therapy. AB - Photodynamic therapy (PDT) is being utilized in the treatment of a wide variety of malignant tumors. Results using PDT have been encouraging, and controlled clinical trials are currently being performed. The procedure exploits both the tumor-localizing and -photosensitizing properties of hematoporphyrin derivative or its purified component, Photofrin II. When this porphyrin mixture is administered systemically, it is retained preferentially in tumor tissue as compared to surrounding normal tissue. Localized tumor destruction induced by PDT results from the photochemical generation of cytotoxic oxygen species within the tumor. This review will provide a summary of historical and current research pertaining to molecular, cellular, and tissue responses induced by PDT. Emphasis is placed on information related to the chemistry of current photosensitizers, subcellular targets, preclinical treatment parameters, and clinical responses following PDT. PMID- 2976444 TI - Percutaneous transluminal excimer laser angioplasty in total peripheral artery occlusion in man. AB - Laser angioplasty and laser-assisted angioplasty have become a clinical reality. Producing sharply defined borders of the ablated area with minimal adjacent thermal damage, excimer lasers offer several proven and some potential advantages over conventional systems. To evaluate the feasibility of excimer laser angioplasty, we have treated one patient using 308-nm radiation via a bare fiber in direct contact with the total occlusion of a right femoral artery. The lesion was successfully recanalized, thus allowing easy passage of the balloon catheter and subsequent dilatation. This percutaneous laser recanalization of an occluded peripheral artery is one of the first to be done in man using excimer laser radiation, thus demonstrating that the technique is feasible and the system is potentially useful. PMID- 2976445 TI - Histopathology of human laser thermal angioplasty recanalization. AB - Laserprobe thermal-assisted balloon, angioplasty (LTBA) has demonstrated promising initial clinical results in recanalizing stenotic or occluded superficial femoral and popliteal arteries. Over the past year we have obtained six specimens of laserprobe thermal (LT) and LTBA treated total occlusions (avg. length 12 cm) for histopathologic examination from patients who were treated for limb salvage. Three tissue specimens were obtained acutely, and one was obtained at 6, 8, and 13 days, respectively, after laser angioplasty at the time of revision for complications or failed procedures. Serial histologic sections of the treated LT segments demonstrated recanalization of atherosclerotic lesions to approximately 60-70% of the probe diameter. The LT channels were lined by a thin layer of carbonized or coagulated tissue and several layers of cell necrosis. The histology of the thermal injury was similar regardless of whether it was produced by the heated metal cap or by free argon laser energy. Stellate balloon angioplasty fractures were frequently filled with thrombus. Analysis of these human LT and LTBA specimens revealed that the thermal device produces a confined injury through the path of least resistance. Balloon dilatation produces fragmented cracks in the vessel wall, which appear to be more thrombogenic than the carbonized LT surface. With improved guidance methods, LTBA shows potential for continuing development. PMID- 2976446 TI - Plaque-media rewelding with reversible tissue optical property changes during receptive cw Nd:YAG laser exposure. AB - Laser dosimetry for thermal fusion of plaque-wall separations during laser balloon angioplasty (LBA) is dependent upon the optical properties of the atheromatous arterial wall during one or more exposures to cw Nd:YAG laser radiation. An integrating sphere technique was used to measure relative transmission and reflection continuously during irradiation of human postmortem atheromatous aortic sections. Tissue luminal surface temperature was recorded continuously with a thermographic video imager during repetitive 20-30-sec, 8-15 watt exposure of a 3-mm nominal spot. In all specimens, transmission fell progressively during each exposure by 10-70% of baseline values. This effect was reversible with normalization of transmission during the initial phase of each subsequent exposure. Changes in transmission were inversely related to temperature over a 50-170 degrees C range, whereas relative reflection remained constant. Accompanying reversible transmission changes was the observation that the weld strength of plaque-aortic wall separations was unchanged by repetitive laser welding and tissue separation of individual sections. In conclusion, temperature-dependent reversible optical and physical properties of plaque occur during exposure to 1.06 microns cw laser radiation. PMID- 2976447 TI - Photodynamic assay of light distributions in tissue phantoms. AB - The distribution of light in tissues of varying blood content has been modeled using a gel agar system as a tissue phantom. Hematoporphyrin derivative (Photofrin I or HpD) was suspended in gels, along with a chemical indicator of its cytotoxic intermediate (singlet oxygen), and various amounts of red blood cells. The singlet oxygen detector used was tryptophan and its oxidation product. Qualitative analysis of the tryptophan photoproduct was determined using high pressure liquid chromatography analysis of individual gel sections. The relative amount of photoproduct was used as a measure of the fluence of actinic light at a given depth within the tissue phantoms. In this manner, the relative activation efficiencies of HpD were determined for light from an argon laser (mainly 488 and 514.5 nm) and an argon-pumped dye laser (630 nm). PMID- 2976448 TI - Technique for measurement of one-dimensional instantaneous ablation velocity. AB - An oscillating mirror scanned a He-Ne laser beam along the length of a thin rod while the rod was ablated by an argon laser beam. The shadow of the scanning beam on an image plane provides a picture of the instantaneous ablation velocity of the rod. This configuration provides a unique method for investigating one dimensional ablation. PMID- 2976449 TI - The concept of nomogenic disorders. PMID- 2976450 TI - The relationships between oligomeric structure and function of band 3 protein from human erythrocyte membranes: present knowledge and suggestions for further experiments. PMID- 2976451 TI - [High professionalism--a pledge of success]. PMID- 2976452 TI - [The 1st All-Union Congress of Physicians]. PMID- 2976453 TI - [The prophylactic work of the nurse in a pediatric polyclinic in protecting child health]. PMID- 2976454 TI - Regulation of Ca2+-pump from cardiac sarcoplasmic reticulum. PMID- 2976455 TI - Kinetic and equilibrium characterization of an energy-transducing enzyme and its partial reactions. PMID- 2976456 TI - Approaches to studying the mechanisms of ATP synthesis in sarcoplasmic reticulum. PMID- 2976457 TI - Fluorimetric detection and significance of conformational changes in Ca2+-ATPase. PMID- 2976458 TI - Synthesis of ATP from Ca2+ gradient by sarcoplasmic reticulum Ca2+ transport ATPase. PMID- 2976459 TI - Occluded Ca2+. PMID- 2976460 TI - Chemical derivatization of Ca2+-pump protein from skeletal muscle with N substituted maleimides and 5-(2-iodoacetamidoethyl)aminonaphthalene 1-sulfonate. PMID- 2976461 TI - Isolation and characterization of sarcolemmal vesicles from rabbit fast skeletal muscle. PMID- 2976462 TI - Use of detergents to solubilize the Ca2+-pump protein as monomers and defined oligomers. PMID- 2976463 TI - Analysis of two-dimensional crystals of Ca2+-ATPase in sarcoplasmic reticulum. PMID- 2976465 TI - Purification, reconstitution, and regulation of plasma membrane Ca2+-pumps. PMID- 2976464 TI - Reconstitution of calcium pumping of cardiac sarcoplasmic reticulum. PMID- 2976466 TI - Isolation of sarcoplasmic reticulum fractions referable to longitudinal tubules and junctional terminal cisternae from rabbit skeletal muscle. PMID- 2976467 TI - Purification of phospholamban from canine cardiac sarcoplasmic reticulum vesicles by use of sulfhydryl group affinity chromatography. PMID- 2976468 TI - Isolation of the junctional face membrane of sarcoplasmic reticulum. PMID- 2976469 TI - Isolation of triads from skeletal muscle. PMID- 2976470 TI - Genetics of Kdp, the K+-transport ATPase of Escherichia coli. PMID- 2976471 TI - K+-ATPase from Escherichia coli: isolation and characterization. PMID- 2976472 TI - Isolation of transverse tubule membranes from skeletal muscle: ion transport activity, reformation of triad junctions, and isolation of junctional spanning protein of triads. PMID- 2976473 TI - [Current epidemiology of Central Asiatic tick-borne borreliasis (based on the example of the Fergana region)]. PMID- 2976474 TI - [Opisthorchiasis in the Moscow and Vladimir regions]. PMID- 2976475 TI - [Opisthorchiasis in the Upper Kama basin]. PMID- 2976476 TI - [Isolation of the Tyagin virus from Anopheles pulcherrimus Theob. mosquitos in Tajikistan]. PMID- 2976478 TI - [Role of non-enzymatic glycosylation of proteins in the pathogenesis of diabetic complications]. PMID- 2976477 TI - [Non-enzymatic glycosylation of proteins in diabetic disease]. PMID- 2976479 TI - An improved method for the isolation of rat cardiac sarcoplasmic reticulum. AB - Preparations of cardiac sarcoplasmic reticulum (CSR) isolated from the rat by differential centrifugation have been widely used for measuring alterations in intracellular calcium flux in response to metabolic and pharmacologic disruptions. However, the purity of these SR fractions has not been firmly established. Using a combination of differential and linear sucrose gradient centrifugation, we have isolated rat CSR with high specific activity and purity. By SDS-PAGE analysis, the preparation is enriched in a protein (110 kD) of similar size to the Ca2+-ATPase of SR from other sources. Gels stained with the dye 'Stains-All' reveal a blue colored 55 kD band, confirming the presence of calsequestrin, the intraluminal low-affinity calcium binding protein of SR. The presence of the transmembrane 53 kD glycoprotein of SR was confirmed by endoglycosidase-H treatment followed by SDS-PAGE and also by a modified Western blotting technique. The rate of calcium uptake in this preparation averages 130 nmol/mg over the first minute of accumulation, approximately 4 times that previously reported for rat CSR. Calcium uptake in our preparation was essentially complete within 5 minutes. Preparations isolated by this method should be of value in future studies measuring alterations in rat CSR function. PMID- 2976480 TI - [Heterogeneity of globin protein synthesis in bone marrow cells of patients with homozygous beta-thalassemia from Tadzhikistan]. AB - The synthesis of globin proteins in blood reticulocytes of homozygous beta thalassemic patients from Tadzhikistan has been previously studied. beta thalassemia with sharp repression of beta-globin protein synthesis (alpha/beta greater than 10) has been shown to be most representative for the region. In this work, the synthesis of globin proteins has been studied in bone marrow cells of homozygous beta-thalassemic patients. Comparison of data on globin synthesis in bone marrow cells and in blood reticulocytes of the patients has revealed that in some cases the disbalance of chain synthesis in both cell types is equal. In other cases the disbalance in bone marrow cells is less than in blood cells, indicating the instability of beta-globin mRNA that is partially degrading in the process of cell maturation. Homozygous beta-thalassemic cases with low content of Hb F in blood cells (5-10%), with substantial disbalance of alpha and beta-globin synthesis and marked production of gamma-globins in bone marrow cells and in blood reticulocytes are of special interest. It has been assumed that parallel to beta-thalassemia some instability of gamma-globin proteins takes place in these patients. PMID- 2976481 TI - [Reabsorption of sutures of the abdominal wall]. PMID- 2976482 TI - [Acute surgical abdomen and cryoglobulinemic purpura. Considerations apropos of a treated case]. PMID- 2976483 TI - Cell differentiation and pattern formation in the developing mammalian retina. PMID- 2976484 TI - [Effect of the spleen cells from partially splenectomized and partially hepatectomized mice on the proliferation of hepatocytes in the regenerating liver of syngeneic recipients]. AB - Cells of mouse spleen obtained 48 h after foster splenectomy, foster hepatectomy, resection of 2/3 of spleen or 36 h after resection of 2/3 of liver were introduced intravenously into partially hepatectomized (resection of 2/3 or 1/4 of liver) syngeneic recipients. Cells of regenerating spleen sharply inhibited the mitotic activity of cells of the recipient liver following resection of 1/4 of liver 48 h after the operation and introduction of cells. Inhibition proved to be dose-dependent: it became apparent when 30 million cells were introduced, increased at a dose of 60 million cells and remained at the same level at higher doses. Division of hepatocytes after resection of 1/4 of liver was inhibited by spleen cells taken in the donors 36 h after partial hepatectomy. Spleen cells of intact and pseudo-operated donors had no such ability. Introduction of 60 million of cells of the regenerating spleen and of the spleen of partially hepatectomized animals into recipients with resection of 2/3 of liver did not inhibit reliably the division of hepatocytes, thus indicating the dependence of inhibition on the level of suppressors in the organism. Resection of a major part of liver was accompanied by a greater decrease in the activity of endogenous suppressors which could not be recovered by the introduced cells. Inhibition of cell division by suppressors was not organ specific. Suppressors inhibited proliferation in liver irrespective of the site of operation. PMID- 2976485 TI - [Myosin isoforms of the developing skeletal muscles in the loach]. AB - Changes in the myosin isozyme spectrum were studied in the loach developing skeletal muscle. It was shown using disk-electrophoresis in polyacrylamide gel and peptide mapping that light and heavy myosin chains from the larval muscles, as well as from the red and white muscle of adult fish differ from each other. Forms of myosin light and heavy chains were found which were characteristic of the larval muscle only. PMID- 2976487 TI - The role of the pediatrician in the care of the handicapped child. PMID- 2976486 TI - Cellular immune status in retinitis pigmentosa. AB - Previous reports have suggested a role of cellular and/or humoral immunity in retinitis pigmentosa. Because of the controversial nature of many of these reports, the authors undertook a detailed investigation of cellular and humoral immunity in a well-characterized group of 47 persons with retinitis pigmentosa of various heritability patterns and a similar number of age- and sex-matched controls. The authors found two changes in lymphocyte subsets. Retinitis pigmentosa patients had significantly elevated Leu 3A-positive lymphocytes (CD4 or T-helper cells) and significantly fewer Leu 2A-positive lymphocytes (CD8 or T suppressor cells) than controls, although the total numbers of T cells did not differ between the two groups. A small but significant number of retinitis pigmentosa patients expressed interleukin 2 (IL-2) antigens on their lymphocytes as compared with none of the controls. The authors saw no differences between the retinitis pigmentosa and control groups in the inducibility and secretion of gamma-interferon or IL-2. Concentrations of immunoglobulins G, A, and M did not differ between the two groups. The link between immune system alterations and the retinitis pigmentosa process remains tenuous. PMID- 2976488 TI - Psychological problems of disabled adolescents and young adults. PMID- 2976489 TI - A developmental perspective for the rehabilitation of children with physical disability. PMID- 2976490 TI - Enhancing the coping of young children with disabilities. PMID- 2976491 TI - Combined passive-active immunization against the hepatitis B virus of 132 newborns of chronic carrier mothers: long term results. AB - One hundred thirty-two newborns of carrier mothers were given combined passive active immunization against the hepatitis B virus. Doses of hepatitis B immunoglobulin (HBIG) were 0.5 ml, and doses of vaccine (Heptavax B) were 10 micrograms. Schedules used were: HBIG, 0, 3 and 6 months and vaccine 1, 2 and 7 months (Schedule 1); HBIG, 0, 3 and 6 months, vaccine, 7, 8 and 13 months (Schedule 2); HBIG, 0 month and vaccine 1, 2 and 7 months (Schedule 3); HBIG, 0 month and vaccine 0, 1 and 6 months (Schedule 4). The study population consisted of 60 boys and 72 girls. Ethnic origins were the following: Haitian, 42; Cambodian, 33; French Canadian, 22; Vietnamese, 21; others, 14. Thirty-two children were born to hepatitis B e antigen-positive mothers. General outcome was: 121 became anti-hepatitis B surface antigen-positive; 5 became hepatitis B surface antigen-positive; and 6 remained negative for all markers (nonresponders). Sex, ethnic origin and immunization schedule had no influence on the outcome. Of the 32 children born to hepatitis B e antigen-positive mothers, only 4 became hepatitis B surface antigen-positive. In those who became anti hepatitis B surface antigen-positive, mean serum anti-hepatitis B surface antigen antibody concentrations (log mIU/ml +/- SD) were: 3.47 +/- 1.03 at 8 to 9 months post-Vaccine 1; 2.89 +/- 0.91 at 14 to 17 months; 2.39 +/- 1.0 at 23 to 29 months; and 2.26 +/- 0.92 at 35 to 41 months.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976492 TI - [Isolated metrorrhagia in a prepubertal child caused by functional ovarian cyst]. AB - The authors report the case of a 10-year-old girl, without any sign of pubertal development, who showed repeated metrorrhagia, related to a functional ovarian cyst. Cyproterone acetate therapy gave only a transient remission of the metrorrhagia. A complete remission was obtained with a 6 months treatment with an LHRH analog. PMID- 2976493 TI - Postoperative erythrodermia (POED), a type of graft-versus-host reaction (GVHR)? AB - Postoperative erythrodermia (POED) is a rare disease appearing several days after surgery for which blood transfusion has been required, characterized by erythrodermia, fever, pancytopenia, hepatic insufficiency or diarrhea. Most affected patients die within four weeks. The etiology remains unknown, but some assume it to be a type of graft-versus-host reaction (GVHR) caused by transfused lymphocytes. In this study, skin biopsies taken from 9 POED patients were examined to clarify whether POED is a type of GVHR. In seven samples, changes compatible with GVHR, such as lymphocyte infiltration in the basal layer and occasional eosinophilic or vacuolar degeneration of the epidermal cells, with occasional satellitosis, were noted. Immunopathologically, infiltrating lymphocytes mostly expressed suppressor/cytotoxic T cell markers like those in GVHR. Meanwhile, immunostaining with anti-HLA-A type specific antibodies failed to differentiate infiltrating lymphocytes from host cells. Thus, the present morphologic and in vivo marker study suggested POED to be a type of GVHR, while in vivo HLA-A typing showed two contradictory possibilities: first, that POED is a GVHR in which major histocompatibility antigen-matched donor lymphocytes attack host cells, and secondly that POED is not a GVHR, the infiltrating lymphocytes being of host origin. PMID- 2976494 TI - Follicular mucinosis of childhood and adolescence. AB - We cared for nine patients who were less than 21 years old at the time of diagnosis of follicular mucinosis. Four had clearing of the lesions, four did not, and one was lost to follow-up. Follicular mucinosis, even in childhood, may be persistent and be associated with lymphoma, which can result in death. Because there is no clinical pattern or single histopathologic criterion that correlates with the outcome, clinical follow-up and assessment of several histopathologic criteria must be relied on when evaluating this condition in children. PMID- 2976495 TI - Etretinate in severe psoriasis of children. AB - Ten children, five with generalized pustular psoriasis and five with erythrodermic disease, were treated with etretinate and observed for up to seven years. In all those with pustular psoriasis, complete clearing was obtained in three weeks to four months, but in one child maintenance therapy had to be introduced twice, for one year each time. In those with erythrodermic psoriasis, the results were favorable only in two, and in two it was necessary to introduce Re-PUVA. Clinical side effects were slight, and laboratory examinations did not disclose any significant abnormalities. In one child who was treated intermittently for seven years, focal osteoporosis of one tibia was disclosed by radiography. No adverse effects on children's development were noticed. PMID- 2976496 TI - Percutaneous absorption of hydrocortisone during and after the acute phase of dermatitis in children. AB - Percutaneous absorption of hydrocortisone was measured by a four-hour absorption test in 16 children during and after the acute phase of dermatitis. In the acute phase, after the application of 1% hydrocortisone cream the increment in plasma cortisol ranged from 47 to 961 nmol per liter (median 248 nmol/L). After the acute phase the increment was significantly (P less than 0.01) lower: range 18 to 241 nmol per liter (median 95 nmol/L). In addition to local effect, topical hydrocortisone therapy of childhood dermatitis is accompanied by a systemic effect that may be pronounced in the acute phase of disease. Convalescence is connected with a diminished systemic effect due to the restoration of the skin barrier to hydrocortisone. PMID- 2976497 TI - Fixed drug eruption at the site of BCG vaccination. PMID- 2976498 TI - [Improvement in the organization of the detection of extrapulmonary tuberculosis in ambulatory care conditions]. PMID- 2976499 TI - [Consultative work of a tuberculosis specialist in a therapeutic hospital]. PMID- 2976500 TI - [The value of an epidemiologic study in detecting sources of tuberculosis infection]. PMID- 2976501 TI - [Experience in the work of detecting tuberculosis in Northern nationalities]. PMID- 2976502 TI - [History of the Chair of Phthisiology at the Tomsk Medical Institute]. PMID- 2976503 TI - [Detection of tuberculosis and nonspecific lung diseases in chemical industrial enterprises]. PMID- 2976504 TI - [Social and vocational rehabilitation in the complex treatment of newly registered patients with active tuberculosis of the respiratory organs]. PMID- 2976505 TI - [Value of complex expeditions for taking intensive anti-tuberculosis actions among children and adolescents]. PMID- 2976506 TI - Carbon-14 labeling and biological activity of the tumor-localizing derivative of hematoporphyrin. PMID- 2976507 TI - The effect of liposomes' membrane composition on the binding of the photosensitizers Hpd and photofrin II. PMID- 2976508 TI - The extrasynaptic effect of gamma-aminobutyric acid and pentobarbital and the influence of temperature on the response of parallel fibres of the frog cerebellum in vitro. AB - The authors studied the effect of two biologically active substances (gamma aminobutyric acid-GABA, pentobarbital-PB) and a physical factor (temperature-T) on the direct response of parallel fibres of the isolated frog cerebellum to electrical stimulation in vitro. The extrasynaptic action of GABA and PB during superfusion (10(-6), 10(-5), 10(-4) and 10(-3) mol.l-1) significantly reduced the amplitude of the response of parallel fibres. Superfusion with picrotoxin (10(-6) mol.l-1) only partly blocked the effect of GABA (10(-3) mol.l-1), although it abolished the effect of PB (10(-3) mol.l-1). Cooling the cerebellum from the control temperature (T = 16 degrees C) to T = 13 and 10 degrees C significantly augmented the amplitude of the responses, while raising it to 19 and 22 degrees C significantly reduced their amplitude. At T = 13 degrees C, depression of direct responses was significant only in superfusion with GABA (10(-6) and 10(-3) mol.l 1) and not in superfusion with PB (10(-6) and 10(3-) mol.l-1). The results with picrotoxin (PTX) applications, indicated that the extrasynaptic action of GABA and PB took effect by partly different mechanisms. That would account for the difference in the effect of GABA and PB in conjunction with the physical factor. PMID- 2976509 TI - Effect of dietary energy intake on tubular reabsorption of urea in sheep. AB - The aim of the experiment was to determine the effect of dietary energy intake on renal urea excretion in sheep with different nitrogen intakes. The control sheep, with a high nitrogen and energy intake, were given a daily feed dose of 21.18 g N and 15.2 MJ digestible energy (DE). The two experimental groups, with an equal, low nitrogen intake, were given diets with a different energy content. The high energy diet contained 3.63 g N and 14.18 MJ DE, the low energy diet 3.4 g N and 6.44 MJ DE. After nine weeks' adaptation to the diets, renal functions were measured by a standard clearance technique. It was found that, under stable urine flow conditions, both groups given the low nitrogen diet had a significantly lower glomerular filtration rate, fractional urea excretion and total urea excretion. A reciprocal comparison of these two groups showed that fractional urea excretion by the sheep with a high energy intake was significantly lower than in the group with a low energy intake. There were no differences in the glomerular filtration rate. A raised dietary energy intake in the presence of a low nitrogen intake caused marked natriuresis and kaliuresis. The results indicate that a raised dietary energy intake can be a significant factor in potentiating the renal effect of urea retention in sheep with a low nitrogen intake. PMID- 2976510 TI - Comparison of the ulcerogenicity of indomethacin and the gastric secretion in verapamil-treated rats. AB - The authors studied the effect of different doses of verapamil on the ulcerogenic activity of indomethacin (20 mg.kg-1) in rats. This was compared with the effect of verapamil on total gastric juice secretion, the amount of acid and the pH. It was found that, as distinct from total secretion and the amount of HCl, which verapamil reduced in correlation to the dose, ulcerogenicity after indomethacin was inhibited the most by a dose of 10 mg.kg-1 verapamil. Larger doses (20 and 30 mg.kg-1) did not increase the anti-ulcerogenic effect any further. This implies that verapamil-induced inhibition of the ulcerogenicity of indomethacin is not related directly to inhibition of total and acid gastric juice secretion. PMID- 2976511 TI - Proceedings of the Czechoslovak Physiological Society. Prague, February 2-4, 1988. Abstracts. PMID- 2976512 TI - Acute low back pain: diagnosis and management of mechanical back pain. AB - Low back pain is the most common musculoskeletal complaint seen in an ambulatory care setting and affects 80 per cent of all persons at some time in their lives. It is the number one cause of activity restriction in people under 45. With the exception of progressive neuromotor compromise, the initial treatment of nearly all causes of LBP is conservative: the use of analgesics and/or anti-inflammatory agents, bed rest, and patient education. PMID- 2976513 TI - [Hyperestrogenism in swine due to natural poisoning with zearalenone]. AB - A field case is described in which all prepuberal swine of a group of 20 pigs and 11 sows showed marked estrogenic effects. These consisted of enlarged mammary glands, swelled tumefacient vulva, and greatly enlarged internal reproductive organs. The corn used to feed these animals was found to contain 56 ppm zearalenone. Deoxynivalenol (4.9 ppm) was found in the corn; T-2 toxin, nivalenol, fusarenon-X, diacetoxyscirpenol, aflatoxins and ochratoxins were absent. Identity of Z was confirmed by TLC in four solvent systems, behavior of the suspected spots under UV light of different wavelengths, change of fluorescence from green to blue after spraying with 5% AlCl3 in alcohol and heating at 110 degrees C during 5 minutes, and by its UV spectrum. A zearalenone producing strain of Fusarium oxysporum was isolated from the suspected grain. Histopathology of uterine tissue showed typical changes produced by zearalenone: hyperplasia, hypertrophy, and metaplasia of the myometrium. Feeding of the grain to a prepuberal sow under controlled conditions reproduced all the effect found in the farm animals. This is the first field case of zearalenone poisoning reported in Argentina. PMID- 2976514 TI - [Clinical management of a case of epidermolysis bullosa]. PMID- 2976515 TI - Influence of indomethacin on experimental heterotopic bone formation in rats. PMID- 2976516 TI - [Hematoma of the rectus abdominis muscle as false acute abdomen (presentation of 3 cases)]. PMID- 2976517 TI - Effect of atrial natriuretic peptide on arterial pressure and renal function in cirrhotic rats with ascites. AB - Glomerular filtration rate, urine volume, sodium excretion and mean arterial pressure were measured in 10 rats with Cl4C induced cirrhosis presenting sodium retention and ascites, and in 10 control rats before and during the iv administration of the 28 aminoacid rat alpha-Atrial Natriuretic Peptide (alpha ANP) (a bolus of 1 microgram followed by a constant infusion of 33 ng/min). alpha ANP induced a similar increase in glomerular filtration rate and filtered sodium load in both groups of rats. In contrast, the increase in urine volume and sodium excretion produced by alpha-ANP was significantly lower in cirrhotic rats (from 13.8 +/- 1.9 to 37.9 +/- 9.1 microliters/min., and from 0.5 +/- 0.1 to 3.3 +/- 1.0 microEq/min) than in control animals (from 14.6 +/- 1.3 to 102.5 +/- 17.7 microliters/min., p less than 0.005; and from 1.0 +/- 0.3 to 14.1 +/- 3.2 microEq/min., p less than 0.001). The results indicate that in rats with experimental cirrhosis and ascites there are blunted diuretic and natriuretic responses to alpha-ANP, probably as a consequence of the exaggerated tubular sodium reabsorption present in these animals. PMID- 2976518 TI - Massive bilateral retinal vascular occlusion secondary to thrombotic thrombocytopenic purpura. AB - An 18-year-old woman developed sudden diffuse bilateral retinal vascular occlusive disease secondary to thrombotic thrombocytopenic purpura. This microangiopathic disease resulted in massive bilateral capillary closure and severe visual loss. Visual loss occurred while responding to therapy for this immune-mediated disease. This report is the first description of extensive retinal vascular occlusion in thrombotic thrombocytopenic purpura. PMID- 2976520 TI - [Immunopathology of the labial salivary glands in Sjogren's syndrome and other autoimmune diseases]. AB - 73 minor salivary gland biopsies (lip biopsy) were performed in patients with primary and secundary Sjogren's syndrome and other collagen diseases without sicca syndrome were studied by means of immunohistochemistry. The results were compared to those obtained in sarcoidosis, sialadenitis and normal glands. In all cases of Sjogren's syndrome and of other sialadenites, we could note predominance of T-lymphocytes with an increase of T-Helpers and a T-suppressor ratio superior to 1. Besides, HLA-DR antigen was present in all the activated cells of lymphocytic infiltrates, in endothelial cells and in some epithelial ductal and acinous cells around the infiltrates. This method cannot permit to differentiate Sjogren's syndrome from the other sialadenites. But, it is very useful to appreciate the evolutivity of the disease (increase in B-lymphocytes in the initial period and in evolutive phases) and to detect its change into pseudolymphoma (polyclonal cells with increase in B-cells) and into malignant lymphoma (usually monoclonal B-lymphocytes proliferation). PMID- 2976519 TI - Effects of exercise on atrial natriuretic factor. Release mechanisms and implications for fluid homeostasis. AB - Atrial natriuretic factor is reported to be elevated during and immediately following exercise and is thought to play a role in fluid homeostasis and cardiovascular regulation. The predominant stimuli for atrial natriuretic factor release during exercise appear to be increases in atrial pressures or atrial distension, both of which are reported to increase with exercise. The intensity and perhaps duration of exercise also influence the magnitude of the atrial natriuretic factor response. It is not clear if the rise in plasma atrial natriuretic factor during exercise plays any role in altering renal function since high intensity exercise is typically associated with an antidiuresis. However, elevations in plasma atrial natriuretic factor may in part be responsible for the increase in urine flow reported when exercise is performed at low or moderate intensities. Atrial natriuretic factor also has vascular effects which may be important in buffering or moderating the blood pressure response to exercise. The atrial natriuretic factor response to exercise and basal levels of the hormone are greatly elevated in patients who suffer from a variety of cardiovascular and pulmonary disease conditions. These elevated plasma atrial natriuretic factor values are associated with increases in atrial pressure and appear to be related to the severity of disease. Although much controversy exists regarding the renal and vascular effects of atrial natriuretic factor, the measurement of this hormone, particularly during exercise, may be of clinical value by providing an additional tool to evaluate patients and determine the effectiveness of various treatment regimens. PMID- 2976521 TI - Inhibitory effects of histamine on interleukin 2 and gamma interferon production of different human T helper cell subsets. AB - We have previously demonstrated that histamine can inhibit human helper T cells by direct interaction with these cells. It has now been investigated whether histamine inhibits lymphokine production by various subsets of CD4+ human T cells separated with the Leu-8 (p80) and Leu-18 (anti-CD45R;p220) monoclonal antibodies (MoAb). Histamine was shown to suppress to a similar extent the production of interleukin 2 (IL-2) and gamma interferon (IFN-gamma) by Leu 3+, Leu 3+8+, and Leu 3+8- cell subsets. Mitogen-activated, unseparated Leu 3+ and purified Leu 3+8 cells produced maximal amounts of IL-2 after 24 h and IFN-gamma after 72 h of culture. In contrast, the Leu 3+18+ subset produced no IL-2 after 24 h, and maximal amounts of IL-2 no sooner than 48 h of culture, and only small amounts of IFN-gamma during the entire culture period of 96 h. Histamine suppressed the production of IL-2 by both subsets, both when produced early (after 24 h), as in the case of the Leu 3+18- subset, and late (after 48 h of culture), as for the Leu 3+18+ subset. The IFN-gamma production by the Leu 3+ and Leu 3+18- cells and the marginal production by Leu 3+18+ cells were significantly suppressed by histamine. Dual staining with Leu 8 and Leu 18 MoAb demonstrated that the Leu 18- cell compartment included both Leu 8+ and Leu 8- cells. It was shown that the inhibitory effect of histamine on the early production of IL-2 and the major production of IFN-gamma by T helper cells is mediated via action on both the Leu 3+18-8- and the Leu 3+18-8+ cells. The inhibitory effect of histamine on the late production of IL-2 is mediated mainly via action on Leu-18+ cells. PMID- 2976522 TI - Tolerogen-mediated suppression of the immune response. Suppressor cells as passive transfer agents. AB - The anti-dinitro-phenyl (Dnp)IgM antibody response in mice was inhibited by administration of either a non-immunogenic form of Dnp-polyacrylamide (Dnp-Pa) or an excess amount of an immunogenic form of Dnp-Pa. Spleen cells, alive or heat killed, from mice tolerized in vivo by either method, inhibited the anti-Dnp response of naive spleen cells co-cultured in vitro with antigen. Conversely, donor cells tolerized in vivo by a high dose of immunogenic Dnp-Pa, when titrated into a naive cell culture which contained no antigen, produced a stimulatory dose response curve. Both the dose-dependent inhibition and stimulation correlated strongly with the amount of 125I-labelled Dnp-Pa carried over by tolerized spleen cells into the naive cell culture system. Because the doses and experimental procedures used were comparable to those commonly used for suppressor cell generation and assay, it is suggested that antigen-specific suppressor cells may produce their effects through passive transfer of antigen and/or tolerogen. PMID- 2976523 TI - Synovial dendritic cells and T cells in rheumatoid arthritis. PMID- 2976525 TI - Quality of life and evaluation of functions among people with severely impaired mobility and non-disabled controls. PMID- 2976524 TI - Occurrence of spinal pain syndromes in a group of apparently healthy and physically fit sportsmen (orienteers). AB - The prevalence of three spine-related pain syndromes was assessed in 739 apparently healthy leisure-time sportsmen (orienteers). The three syndromes were (i) low back pain or stiffness, (ii) thoracic inter-scapular back pain or stiffness, and (iii) frontal chest pain or discomfort. The mean age of the sportsmen was 32.7 years (range 10-65). For low back pain or stiffness the cumulative occurrence was 471 per 1,000, for thoracic back problems it was 146 per 1,000, and for chest pain or discomfort 85 per 1,000. The three pain syndromes were highly associated, i.e. they tended to occur in the same individual. PMID- 2976526 TI - Low back pain in schoolchildren. An epidemiological study. AB - A prospective epidemiological study was conducted to determine the prevalence of backache and the aetiological role of several contributing factors. 1715 schoolchildren of both sexes answered a previously validated questionnaire. Approximately 33% of our sample had already suffered from low back pain (LBP) at some time. There was a statistically significant positive correlation between LBP and age, female sex, time spent watching TV, smoking and competitive sports. PMID- 2976527 TI - Income redistribution effect of the Swedish sickness allowance insurance in a comparison of two concepts of social class. AB - Social insurances effect income distributions between social strata. Here, insurance returns in relation to income are studied on the Swedish sickness allowance insurance, which is intended to redistribute from higher to lower social strata. Two measures of social class are used, the socio-economic classification, the official index of Sweden, and a structural class concept, which in earlier results discriminates better for material factors such as income and work conditions. The material consists of all sickness cases of 1983 for 3,161 persons, sampled from insurance registers and cross-classified with registers at taxation authorities. Data on insurance returns, incomes, and occupation are used. Results clearly confirm the intended redistribution effect, but considerably clearer with the structural class concept. The effect is even stronger than intended for some strata, where the system seems to lack in implementation. The consequences for choice of class measure are finally discussed. PMID- 2976528 TI - Differential mortality among semiskilled applicants of disability pension. AB - The mortality experience among 4,440 applicants for disability pension has been examined during a period of observation of ten years. All applicants were males and had been member of the Danish Semiskilled Workers' Union. The reference group consists of age matched male members of the same union. The applicants for disability pension experienced a considerable excess mortality when compared with the reference group. The higher mortality risk is still present more than ten years after the application has been settled. The persons who were refused disability pension experienced an elevated mortality, too. In opposition to previous studies it is not found informative to put forward a common estimate of the elevated mortality rate because the group of disability applicants is very heterogeneous concerning their mortality experience; the excess mortality is most elevated among persons granted the high level pension, among the youngest, and during the period closest to the award. PMID- 2976530 TI - [Experience with promoting the human factor in public health]. PMID- 2976529 TI - Chronic recurrent multifocal osteomyelitis and psoriasis--a report of a new association and review of related disorders. AB - In summary, we have described two patients with CRMO and psoriasis, and have reviewed the musculoskeletal manifestations associated with pustular eruptions of the palms and soles. In view of the frequent occurrence of PPP in patients with CRMO, we suggest that the occurrence of psoriasis in our two patients is more than coincidence, and that noninfectious, inflammatory lesions of bone may be another musculoskeletal manifestation of psoriasis. This rare association, as well as the association of PPP with disorders associated with new bone formation, may shed new insights on the relatively common finding of periosteal elevation associated with psoriatic arthritis and the occasional severe juxta-articular osteolytic destructive bone lesions seen in psoriatic arthritis. PMID- 2976531 TI - [Experience with scientific and practical work in regard to studying the health status of the population of the Karelian ASSR during the transition to mass screening]. PMID- 2976532 TI - [Social and hygienic characteristics of patients hospitalized as emergency cases in various categories of cities]. PMID- 2976533 TI - [Social and hygienic aspects of the initial stage of sex behavior in an urban population]. PMID- 2976534 TI - [Comparative evaluation of progress in studies of various student groups]. PMID- 2976535 TI - [Patronage assistance to military institutions in the Vologda district during World War II]. PMID- 2976536 TI - [The role of the Perm Medical Institute in the development of higher medical education]. PMID- 2976537 TI - Reproducibility of two in vivo tests of platelet function. AB - We correlated repeat determinations of platelet aggregate ratios (PAR) and plasma beta-thromboglobulin (BTG) in the right and left arms of thirty-four subjects in order to measure test reproducibility and to evaluate the effects of venipuncture. The reproducibility of PAR on the same blood samples was relatively low but similar to that obtained when the right arm and left arm values were compared, thus indicating little variation secondary to venipuncture. For BTG, the reproducibility on identical blood samples was quite high, but fell significantly when plasma samples from the two sides were correlated. This finding and increased BTG values when obvious blood collection problems occurred, indicated that plasma BTG levels are quite sensitive to venipuncture technique. PMID- 2976538 TI - Late results of prosthetic valve replacement for aortic regurgitation and the prognostic significance of the end-diastolic and regurgitated blood volumes. AB - Between January 1975 and December 1985 214 patients underwent prosthetic aortic valve replacement for isolated aortic regurgitation. Patient follow-up averaged 5.4 years after the operation (range 0.7-10.7 years). Preoperative staging showed 9 patients to be in NYHA stage II, 111 patients in stage III, and 94 patients in stage IV. At follow-up 3 patients complained of increased fatigue, 9 patients remained stable and 167 patients had improved. 103 of these patients were free of symptoms under stress, i.e. NYHA stage I. The hospital mortality was 3.7% (8/214 patients). An additional 26 patients died within an average of 2.8 years. The cumulative 5-year survival rate was 85%; the 10-year survival rate was 81.5%. 10 patients underwent a second operation within an average of 3.4 years. The correlation between the end-diastolic volume and the regurgitated blood volume has prognostic significance. 25 patients with appropriate enlargement of the left ventricle showed a significant decrease of the end-diastolic volume and the roentgenographic heart volume combined with an increase of ejection fraction. None of these 25 patients died from cardiac complications. A control group of 9 patients with myocardial damage showed no significant change in the above parameters. 4 patients in the control group died. We conclude that the relationship of regurgitated blood volume and the left ventricular end-diastolic volume is of prognostic significance for patients with chronic aortic regurgitation presenting with minimal symptoms. PMID- 2976539 TI - [Study of a standard nursing care plan--effects of the standard nursing care plan on a patient undergoing PTCA (percutaneous transluminal angioplasty)]. PMID- 2976540 TI - [AIDS--responsibility of the stomatologist in diagnostics and prevention]. PMID- 2976541 TI - [Oral manifestations in infections with HIV in AIDS]. PMID- 2976542 TI - Synthetic-fibre filters for preventing dracunculiasis: 100 versus 200 micrometres pore size. AB - Filtering of drinking water to remove the copepod intermediate hosts of Dracunculus medinensis is a primary strategy for control of guinea-worm disease. Since filters of different porosities are used, we tested the efficiency of synthetic-fibre filters of 100 microns and 200 microns pore size in removing the various stages of 3 species of copepods from water samples. The 200 microns mesh retained the larger copepodid stages including adults (C III-VI), but permitted passage of smaller copepodids and all naupliar stages. The 100 microns mesh retained all but the earliest naupliar stages (N I-II) which are unlikely to harbour guinea-worm larvae. PMID- 2976543 TI - Syngamosis: two new Brazilian cases and evidence of a possible pulmonary cycle. AB - Cases of syngamosis due to Mammomanogamus laryngeus are reported, with a discussion of diagnostic aspects. A pulmonary cycle for M. laryngeus is proposed, suggested by the appearance of a transient area of homogeneous consolidation of the lung, detected by X-ray. PMID- 2976544 TI - Antiproliferative effect of Hu-interferon-gamma in 674V and J82 bladder carcinoma cell lines. AB - Hu-IFN-gamma was evaluated in regard to the antiproliferative effect on J82 and 647V bladder cancer cell lines. In addition, the IFN-receptors were determined. There was a significant growth inhibition of J82 as well as 647V at low dose Hu IFN-g (1 U/ml). The growth inhibition was significantly higher in 647V than in J82. The binding assay for 125J-Hu-IFN-g revealed 870 and 3,000 binding sites for 647V and J82, respectively, indicating that the antiproliferative effect of Hu IFN-g may not depend on the absolute amount of IFN-receptors, in the two cell lines tested. PMID- 2976545 TI - [Endometriosis of the postoperative scar]. PMID- 2976546 TI - Immunophenotype of pulmonary cellular infiltrates in sheep with visceral caseous lymphadenitis. AB - Pulmonary lesions associated with Corynebacterium ovis were analyzed with an indirect immunoperoxidase staining technique using monoclonal antibodies. The predominant cells in abscess walls and surrounding lung parenchyma were large macrophages which expressed major histocompatibility complex (MHC) class II molecules on their surfaces. T lymphocytes were prominent in the same sites in the naturally occurring lesions, and SBU-T4-positive ("helper/inducer") cells were the major subset of lymphocytes (mean T4/T8 ratio = 3.5). B lymphocytes and granulocytes comprised minor populations of infiltrating cells. These results implicate activated macrophages and MHC class II-restricted T lymphocytes in the pathogenesis of established C. ovis infections in sheep. PMID- 2976547 TI - Treatment of chronic back pain in horses. Stimulation of acupuncture points with a low powered infrared laser. AB - Fourteen horses that could not perform at their expected standards due to chronic back pain of 4 to 48 months duration, and had not obtained lasting improvement from other forms of therapy, were treated by stimulating nine acupuncture points using a low powered infrared laser (300 microW, 904 nm). The treatments were performed weekly, and consisted of stimulating each point for 2 minutes with a pulse frequency of 360 pulses per second. After completion of a mean of 11 treatments, clinical signs of back pain were alleviated in 10 of the 14 horses, there was no change in three, and one was lost to follow-up. Of the 10 horses who were training and competing, four won. One year after treatment was discontinued, 9 of these 10 horses continued to perform at a standard acceptable to the owner. PMID- 2976548 TI - Effect of fascial closure technique on strength of healing abdominal incision in the dog. A biomechanical study. AB - Load-deformation data were used to determine the strength of healing abdominal incisions in dogs at 7, 14, and 21 days after wounding. The breaking energy of paramedian incisions closed by suturing the internal and external leaves of the sheath of the rectus abdominis muscle vs. incisions closed by suturing the external leaf only was similar in the 12 dogs studied. Closure of abdominal incisions by suturing only the external leaf of the rectus sheath provided wound strength comparable to the traditional method. PMID- 2976549 TI - Tissue reaction to suture material in the feline linea alba. A retrospective, prospective, and histologic study. AB - Swelling and inflammation along the incision line were observed after elective ovariohysterectomy in 22 of 66 cats in a retrospective study. In a prospective study of 99 feline abdominal incisions closed with surgical gut, polyglactin 910, or polydioxanone, with and without subcutaneous closure, the least inflammation occurred when the linea alba was sutured with polyglactin 910 and the subcutaneous tissues were not sutured. Histologically, reactions in the linea alba of 12 other cats progressed from purulent to fibromononuclear to fibrous within 14 days after closure with gut, polyglactin 910, and polydioxanone. Microscopic evidence of seroma formation in 9 of 12 animals in which the subcutaneous tissue was not sutured suggested that surgical closure of subcutaneous dead space was indicated. The inflammatory reaction did not appear to be related to any one suture material as reactions were seen with all of them. PMID- 2976550 TI - [Comparative study of the state of health and nutrition of young children from rural and urban areas of the Uzbek SSR]. AB - The study of the physical development including caliperometric investigations was conducted in 2620 infants. The food ingredients and caloric content were studied in the daily ration of 267 infants with premorbid signs. Parameters of the physical development (growth, body mass, chest circumference size, and standards of fat content in per cent) in infants can be recommended as reference ones in the region studied. The physical development of infants depended on proper organization of full value nutrition, both quantitative and qualitative. PMID- 2976552 TI - [Improving the organization of dietary services in a sanatorium-preventorium]. PMID- 2976551 TI - [Therapeutic and preventive diet and various indicators of health and work capacity of workers at the industrial plant "Sintez"]. AB - Anthropometric investigations conducted have shown that 28.1% of the workers have excessive body mass. At the same time energy expenditure of workers at some departments and bays is within the range of 164 +/- 1.6-188.89 +/- 3.5 kcal/h. Natural resistance parameters (lysozyme activity, virus persistence) in the workers of the enterprise were lower than in the control group. Physical working capacity of 15% of the workers was rather low. Optimal providing of the body with ascorbic acid, thiamine and riboflavin was recorded only in 30-40% of the examined workers. Ration N 5 that was given to the workers was characterized by excessive energy value, low content of animal protein and vegetable fat deficiency; the content of simple sugars was 2-fold higher than the value recommended. The components of the ration are not sufficiently balanced, it does not meet the physiological needs of the workers. PMID- 2976553 TI - [Body vitamin allowance of vegetable growers working on sheltered soil]. AB - The providing of hothouse workers with vitamins B1, B2, B6, PP, C and E was comprehensively evaluated basing on the study of their actual nutrition and biochemical parameters of their vitamin status--daily excretion of vitamins and their metabolites with urine, vitamin concentration in the blood serum and activity of erythrocytic vitamin-dependent enzymes with calculation of their activation coefficients. Multiple hypovitaminosis detected in the vegetable growers working in hothouses who received normal amounts of these essential nutrients with food, as well as the relationship noted between the vitamin providing level and the length of working in the hothouse, evidence a possible influence of the occupational factors--the character of work attended by the action of toxic chemical compounds under conditions of the hothouse microclimate. Further investigations should be conducted to determine the vitamin requirements of hothouse workers. PMID- 2976554 TI - [Improved microbiological control over the products of sublimation drying]. AB - The qualitative and quantitative composition of microflora was studied in fruit and vegetable juices, pastilles and sauces of sublimation drying. The main groups of microorganisms causing deterioration of these products have been specified, and microbiological tests necessary for standardization of the products have been established. Indication and identification of a microbe type were conducted by methods used in technical microbiology. It has been found that microbiological contamination in fruit puree and juices dried by sublimation comprises (0.1 6).10(2)/g. Maximum content of microorganisms is (3-3.5).10(3)/g. Qualitative composition of the microflora of the products studied is represented by spore mesophyll aerobes of subtilis-mesentericus type (95-98%), yeast, mold fungi, lactic-acid bacteria and cocci. PMID- 2976555 TI - The activities of the Hungarian Centre for Congenital Anomaly Control. AB - The Hungarian Centre for Congenital Anomaly Control manages a number of surveillance programmes based on the Hungarian Congenital Malformation Registry. Notification of congenital anomalies, which is compulsory, originates exclusively from physicians working in various health institutions and is based on their diagnosis of malformed patients--newborns and infants. In recent years, the total birth prevalence of registered congenital anomalies has exceeded 47 per 1,000 total births. The notified data are critically evaluated and centrally coded according to the unit of notification, the affected individual. Diagnostic accuracy, completeness of notifications and the effect of confounding variables are continuously checked. The purpose of the Case-Control Surveillance System is to obtain etiological information concerning drug consumption, maternal diseases and occupational hazards during pregnancy. The surveillance of mutations through indicator conditions (sentinel anomalies. Down's syndrome and pairs of components of unidentified multiple congenital anomalies) is an attempt to measure the rate and trend of new germinal mutations. The nationwide follow-up of multimalformed infants helps to increase the proportion of identified congenital anomaly entities, to identify new ones, and to detect clusters caused by new environmental teratogenic or mutagenic factors. PMID- 2976556 TI - [The health status of the population at the onset of retirement--results of a total survey in Wurzen]. PMID- 2976557 TI - [Headache caused by functional discordance of the pelvis?]. PMID- 2976558 TI - [Responsibilities and possibilities of pain consultation]. PMID- 2976559 TI - [Chromosomal mosaicism after diagnostic chorionic villi biopsy. Subsequent study of the placenta]. AB - Among 311 diagnostic chromosomal analyses after CVS eleven results (mostly chromosomal mosaicism) did not fit completely with the chromosomal sets found in the fetuses and newborns. In all cases chromosomal mosaicism was restricted to the placenta only. All children born after the diagnosis of a chromosomal mosaicism in the placenta were normal. In six cases the investigation of the placenta after termination of pregnancy or birth was possible. In all those cases the mosaicism could be found within the placenta, however, in some cases in a very low percentage. Although occurrence of chromosomal mosaicism in the placenta can hinder chromosomal diagnosis after CVS, the advantages of the method are not neutralized. PMID- 2976561 TI - [Roentgeno-endovascular dilatation of the brachiocephalic arteries]. AB - The article describes the technique of radiological endovascular dilatation (RED) of stenosed brachicephalic arteries. RED was attempted in 28 arteries (18 subclavian, 7 vertebral, 2 common carotid and 1 internal carotid) in 22 patients. In one patient RED of the third portion of the left subclavian artery was supplemented with endovascular prosthesis--transcutaneous transcatheter introduction at the site of the previous stenosis of a spiral-shaped prosthesis from nitinol (a nickel-titanium alloy). Technically good results of dilatation was achieved in 21 of the 22 patients, clinical effect in 20 patients. Some angiographic criteria of the selection of patients for RED of the brachicephalic arteries are described. PMID- 2976560 TI - [Rehabilitation treatment of patients with sequelae of stroke at a specialized rehabilitation center in Leningrad]. AB - The article reviews a ten-year experience with rehabilitation of post-stroke patients accumulated at specialized in- and out-patient rehabilitation centers. The authors present the principles of structuring the recovery process, as well as the main components of rehabilitation programmes, individual methods, and their combination. The ultimate results of rehabilitation treatment are considerably better than those observed following the traditional chemo- and physiotherapy. PMID- 2976562 TI - 5-fluorodeoxyuridine decreases the effect of arabinofuranosyl-cytosine in undifferentiated human B lymphocytes. AB - An activated lymphocyte population was isolated from tonsils of 3-6 years old children by density gradient centrifugation. The isolated light density (LD) cells were bearing early B lymphocyte markers and were 5-6 times more active in DNA synthesis than the high density (HD) lymphocytes. It was found that about 70% of CdR was deaminated and converted into dTMP. The cells were very sensitive to araC, but the incorporation of [14C] CdR was twice more sensitive to araC than that of [5-3H]CdR. This effect can be explained by the interconversion of araC into araT nucleotide via the CdR interconversion pathway. This suggestion was also supported by the effect of 5-F-UdR, which decreased the inhibition of DNA synthesis caused by araC. 5-FUdR the inhibitor of thymidylate synthase, possibly decreases the interconversion of araC into araTTP, and its action via a smaller dTTP pool of the cells. Our data suggest, that the sensitivity of cells to araC may depend on the capacity of the CdR----TdR interconversion pathway, which, however, changes during the normal differentiation process of lymphocytes. Thus, the sensitivity of different malignant cells to araC is determined by the differentiation stage at which they were arrested during cell transformation. PMID- 2976563 TI - Influence of EDTA on the ATPase activities of plasma membranes from winter wheat roots. AB - Plasma membrane vesicles were purified from roots of winter wheat seedlings (Triticum aestivum L. cv. Marton-vasari-8) by aqueous polymer two-phase partitioning. The ATPase activity of vesicles in the presence of Ca was independent of but in the presence of Mg depended on the EDTA concentration in the ATPase assay. The potassium stimulation and vanadate inhibition of the MgATPase also depended on the EDTA concentration. Consequences of these results are briefly discussed. PMID- 2976564 TI - Contractile properties of chick embryo muscles in development. AB - In this paper we report on the observation of two parameters of embryonic muscles which show the functional activity of myofibrillar ATPase activity of embryonic muscles and superprecipitation (SP) of natural actomyosin. Our results indicate that, during the embryonic period, the myofibrillar ATPase activity and the SP of actomyosin significantly increased, the rate of this increase being different for leg and breast muscles. PMID- 2976565 TI - Siderophore containing 2,3-dihydroxybenzoic acid and threonine formed by Rhizobium trifolli. AB - An iron-binding compound was isolated from ethyl acetate extract of culture supernatant fluid of Rhizobium trifolii AR6 and was purified by iron-exchange chromatography. The compound was characterized by UV and IR. It contained 2,3 dihydroxy-benzoic acid and threonine and was accumulated during stationary phase of growth in iron-deficient media. Synthesis of the siderophore was repressed by FeCl3. In iron limited medium the compound promoted growth of R. trifolii strains. PMID- 2976566 TI - [Our experience with TENS in the treatment of chronic backache]. PMID- 2976567 TI - [The spine's absorptive capacity in patients with backache: evaluation of a diagnostic test]. PMID- 2976568 TI - Percutaneous transluminal angioplasty of iliac and femoral arteries in severe lower-limb ischaemia. AB - Percutaneous transluminal angioplasty was performed 92 times in 86 patients with severe lower-limb ischaemia (40% occlusion), giving rise to rest pain and/or gangrene. The patients were thereafter observed for periods up to 5 years. Criteria for success were appearance of normal groin pulse (iliac angioplasty) or persistent greater than or equal to 0.15 rise in arm/ankle blood pressure index (femoropopliteal angioplasty). The respective technical success rates were 82% and 64%. The complication rate was 10.9%, including 5.4% distal embolization. Patency rates were higher in iliac than in femoropopliteal lesions, in stenotic than occluded vessels, and also when the lesion was shorter than 5 cm and if there was good run-off. Limb salvage exceeded patency by 10% in the iliac procedures and by 15% in the femoropopliteal. Percutaneous transluminal angioplasty is recommended for selected cases of severe lower-limb ischaemia, and should always be considered for limb salvage. Reocclusion does not necessarily imply clinical failure. Technical failures should be included in calculated patency rates in order to document the method's limitations. PMID- 2976569 TI - Intraperitoneal granulomatous foreign body reaction after accidental perforation of the abdominal wall. Case report. AB - After an accidental perforation by a wooden stake of the abdominal wall and distal ileum a 28-year-old man developed an aggressive granulomatous foreign body reaction of the greater omentum with high fever and abdominal pain. The patient was cured by omental resection and prednisone treatment. PMID- 2976570 TI - Adenocarcinoma at the ileo-cutaneous junction following subtotal colectomy for ulcerative colitis. Case report. AB - Adenocarcinoma of the ileo-cutaneous junction is exceedingly rare. We report a case. The clinical presentation and pathogenesis are discussed. PMID- 2976571 TI - Plasmin in subretinal fluid. AB - Proteolytic activity was studied in subretinal fluid from 56 eyes with rhegmatogenous retinal detachment without vitreous or subretinal hemorrhage. Active plasmin (1.0-15.2 micrograms/ml) was found in 33 eyes and plasmin inhibitor complexes in 3 eyes. Plasmin was detected more often in large detachments, but there was no clear correlation with the duration of the detachment or characteristics of the holes in the retina. It seems possible that plasmin in subretinal fluid may enhance release of cells from the pigment epithelium by degrading the extracellular matrix and contribute to the development of proliferative vitreoretinopathy. PMID- 2976572 TI - The effect of different LH-RH agonists on the levels of immunoreactive gonadotropins. AB - The present study compared the effect of D-Ser(tBU)6-des-Gly-LHRH-ethylamide in the form of a nasal spray, D-Trp6-LHRH, which is applied in daily intramuscular injections, and the long-acting preparation, D-Trp6-LHRH, which is given as a monthly intramuscular injection, on the function of the pituitary-ovarian axis. Forty spontaneously menstruating women not belonging to the PCOD group participated in the study, the results of which indicate that the three LH-RH agonists used are similarly effective in their ability to down-regulate the pituitary-ovarian axis. The time required for the appearance of the suppressive effect, as measured by reduced levels of gonadotropins and estradiol as well as by the abolishment of the hypophyseal response to rising estrogens, usually does not exceed 14 days and is never more than 28 days. The immunoreactive LH-reducing effect of the LH-RH agonists was true only if the basic levels of the hormone were higher than 11 mIU/ml. This fact might be particularly significant in patients with polycystic ovarian disease or in perimenopausal women. PMID- 2976573 TI - Induction of ovulation with D-Trp6-LHRH combined with purified FSH in patients with polycystic ovarian disease. AB - Seventeen patients with polycystic ovarian disease (PCOD) and evidence of mild or severe ovarian hyperstimulation syndrome (OHSS) during therapy with CC/hCG, FSH/hCG or hMG/hCG were treated with D-Trp6-LHRH until medical gonadectomy was attained. Under the suppressive therapy with the GnRH agonist (GnRHa) ovulation was induced with FSH/hCG. In 15 out of 17 patients, ovulatory cycles were obtained with this new modality of treatment. Seven patients conceived (3 viable pregnancies and 4 early abortions) after the 1st treatment cycle. Fourteen of the 17 patients demonstrated symptoms of mild OHSS which did not require hospitalization. Only 1 patient developed severe OHSS after the combined treatment. Our results suggest that therapy with GnRHa, especially in its delayed release formulation, is effective for the prevention of severe ovarian hyperstimulation in PCOD patients undergoing treatment with menotropins for the induction of ovulation. PMID- 2976574 TI - Measurement in rehabilitation. PMID- 2976575 TI - The Functional Autonomy Measurement System (SMAF): description and validation of an instrument for the measurement of handicaps. AB - The Functional Autonomy Measurement System (SMAF) is an instrument developed for the measurement of the needs of the elderly and the handicapped. Its elaboration was based on the World Health Organization's classification of impairments, disabilities and handicaps. A functional autonomy rating scale, using a four level measurement scale, quantifies a subject's performance on 29 functions in five sectors of activity: activities of daily living, mobility, communication, mental functions and instrumental activities of daily living. For each function, the evaluator must also estimate available resources to compensate for any identified disability in order to estimate the handicap. The disability and handicap profile obtained is the basis for the prescription of home care or the allocation of chronic care beds. An inter-observer study concluded that the scale is reliable for evaluators from different professions in the community as well as in institutional settings. The instrument is rapid to administer (on average 42 min) and the reliability is not influenced by training. A study of concurrent validity has shown a strong correlation between the disability index obtained by the SMAF and the amount of required nursing-care time. This instrument can be used for clinical purposes and in epidemiological and evaluative research. PMID- 2976576 TI - A Cox regression analysis of risk factors related to Mycoplasma suipneumoniae reinfection in Danish SPF-herds. PMID- 2976577 TI - Distribution of lost-work-time claims for skin disease in California agriculture: 1978-1983. AB - In order to identify California agricultural workers at high risk for occupational skin disease, we evaluated the distribution of 2,722 claims for lost work-time skin conditions reported between 1978 and 1983 by Standard Industrial Classification (SIC) and source of illness. In the overall work force, cases were most frequently attributed to plants (52.1%), chemical exposures (20.4%), and food products (12.5%), but considerable variation occurred in the distribution of cases among the subdivisions of agriculture. The forestry subdivisions (SICs 081, 082, 084, and 085) had the highest rate (given as cases/10,000 employed) of claims for skin disease attributed to plants (53.5), but no cases due to other exposures. For chemical exposures, the highest rate of claims occurred in the horticultural specialities (SIC 018) subdivision (5.1), but this group also had a significant claims rate for plant-related skin disease (15.9). The highest claims rate for cases attributed to food products occurred in the vegetables/melons subdivision (4.7), but this group had claims rates for exposure to plants (3.8) and chemicals (2.8) that were nearly as high. Efforts to evaluate and prevent skin disease in agricultural workers should recognize the complex range of exposures to which this work force is subject. PMID- 2976578 TI - Skin notation in the context of workplace exposure standards. AB - In the establishment of workplace exposure standards, the potential for cutaneous absorption is taken into consideration through the addition of "skin notation" to the relevant substance. In the TLVs Documentation (ACGIH, 1986) dermal lethal dose to 50% (LD50) or human data are the bases for the assignment of "skin notation" to 91 of 168 substances. For the other substances, the "skin" attribution seems to be based on undocumented statements in 24 (14.5%), skin effects in 13 (8%), and analogy in 7 (4%), while in the remaining 33 (20%) any reference is lacking as to the basis for notation of the cutaneous route of entry. Furthermore, since the established "cut-off" value of 2 g/kg is sometimes bypassed when a notation is added or omitted, the use of dermal LD50 is perplexing. Given the relevance of the skin notation for the validation of threshold limit values (TLVs) in the workplace, a full examination and citation of all available scientific data are recommended when establishing the TLV of substances absorbable through the skin. PMID- 2976579 TI - Selective termination of multiple gestations. AB - Twenty-two selective terminations in multiple gestations were performed by a number of different methods. In 17 dichorionic pregnancies there was a successful delivery in surviving singletons or twins. In five monochorionic pregnancies undergoing selective termination there was a successful delivery in only one and a pregnancy loss in the other four. Six of the 18 delivered pregnancies were complicated by premature labor and delivery. Among the several methods used for selective termination, intracardiac potassium chloride injection appears to be the procedure of choice in dichorionic pregnancies. PMID- 2976580 TI - The atrioventricular node of the golden hamster (Mesocricetus auratus): a light and electron microscopic study including immunocytochemistry. AB - The atrioventricular (AV) node of the golden hamster is situated unusually high in the interatrial septum when compared to other species such as the rat. 2 main cell types, characterized by electron-lucent or electron-dense cytoplasm respectively, are found in the node; although both types contain numerous myofilaments these are irregularly arranged and sarcomeric banding is poor. A third variety comparising transitional cells, with features intermediate between the main nodal cells and general atrial myocardial cells, are found at the periphery of the node. Similar electron-lucent and electron-dense cells are also found in the bundle and the mean diameter of bundle cells increases as one passes from the node to the bundle bifurcation. In the node, specific heart granules (SHG) identified by ANP-28 immunoreactivity are found only in transitional cells and even here they are very sparse, unlike general atrial myocytes in which they are plentiful. Numerous nerve varicosities are present throughout the node and bundle and 5-hydroxydopamine (5-OHDA) labelling demonstrates that most of them have features of either noradrenergic or cholinergic terminals; a few non cholinergic, non-adrenergic varicosities are also present. PMID- 2976582 TI - Coverage of chest wall and pelvic defects with the external oblique musculofasciocutaneous flap. AB - The external oblique muscle, its fascia, and overlying skin have comprised flaps that have been used successfully in reconstructive surgery of various defects in the past. This flap provides good coverage for lower and upper chest wall defects to the level of the ipsilateral fourth rib. It provides skin of a similar color and texture and of large quantity. The skin of the flap is innervated; thus, sensation can be maintained. The remaining flat abdominal musculature and fascia maintain the integrity of the abdominal wall after transposition. We demonstrate that the external oblique musculofasciocutaneous flap can also be rotated in an inferior direction to close large hemipelvectomy defects in the absence of the usual ipsilateral thigh musculature. PMID- 2976581 TI - [Histochemical ATPase detection in arterial myocytes with regard to their contractile differentiation]. AB - ATPase activity of arterial medial smooth muscle cells in rabbits at different postnatal age was studied. At earlier periods the reaction was very weak in the media. This was followed by intensification after which it continues to be diffuse. Parallel electron-microscopic studies revealed a contractile differentiation which coexists with the activity of the 2 enzymes. PMID- 2976583 TI - Calcitonin gene-related peptide delays the no-reflow phenomenon in the rat island flap. AB - Calcitonin gene-related peptide (CGRP) is a bioactive neuropeptide with potent vasodilatory properties. The effect of CGRP on the no-reflow phenomenon was studied in rats. Island flaps based on the epigastric vessels were exposed to 11 hours of warm ischemia. CGRP was given as single doses before, before and after, or after the ischemic insults. Pre-ischemic treatment with CGRP increased flap survival at concentrations ranging from 10(-9) mol/L to 10(-7) mol/L. The survival rate of saline and untreated control flaps was 18.4%, calculated on the basis of tissue survival areas. The optimum survival rate after preischemic CGRP treatment was 60.3%, and after both preischemic and postischemic CGRP treatment, 66.3% (p less than 0.005 as compared with controls). Given as a single dose after the ischemic period, CGRP increased flap survival to 45.5% at 10(-7) mol/L (p less than 0.05), but no effect was found at lower concentrations. Apart from free radical scavengers, CGRP is the only agent known to delay the no-reflow phenomenon after a single postischemic dose. PMID- 2976584 TI - The free or pedicled anteromedial thigh flap. AB - The anteromedial thigh flap first described by Song is a septocutaneous artery flap based on the septocutaneous perforator originating from the lateral circumflex femoral vessels and long saphenous vein. The use of this flap for 3 patients who required soft tissue coverage is reported herein. The most important advantage of this flap is that it can be used not only as a skin flap but also as a vascularized fascia graft and fasciocutaneous free flap for the full-thickness defect of the abdominal wall and cranial region. PMID- 2976585 TI - Biochemistry and clinical significance of lipoprotein (a). PMID- 2976586 TI - Atlantoaxial instability and abnormalities of the odontoid in Down's syndrome. AB - Radiography of the lateral neck was performed on all children with Down's syndrome aged 4-15 in the Southern Derbyshire health district (n = 67) and a random selection of adults (n = 94). Atlantoaxial instability, defined as a gap of over 4 mm, was present in seven (10%) children and two (2%) adults. Odontoid hypoplasia, defined as an odontoid peg two standard deviations below the mean in an age matched population, was present in 15 (22%) children and 14 (15%) adults, with accessory odontoid ossicles present in two (2%) and two (3%) respectively. No one was found to have symptoms or clinical signs of spinal cord compression. Atlantoaxial instability was therefore found to be commoner in children than adults. Different programmes of management are suggested, in terms of regular clinical examination for signs and symptoms and by radiographic screening. Particular care should be taken with those who have both atlantoaxial instability and odontoid hypoplasia or accessory ossicles as they are at particular risk of spinal cord damage. PMID- 2976587 TI - [The existence of Elaephora poeli (Vryburg, 1879) Raillet and Henry, 1912 in zebu (bos indicus) and Ituri (Upper Zaire)]. PMID- 2976588 TI - Comparative effects of the calcium antagonist isradipine and some other dihydropyridine derivatives on regional blood flow in anesthetized open-chest dogs. AB - The effects of isradipine (PN 200-110), isopropyl 4-(2,1,3-benzoxadiazol-4-yl) 1,4-dihydro-5-methoxycarbonyl-2,6-dim ethyl-3- pyridinecarboxylate, on some cardiovascular parameters and regional blood flow were compared with those of other dihydropyridine derivatives in anesthetized open-chest dogs. Intravenous (i.v.) administrations of isradipine 3 and 10 micrograms/kg, nifedipine 10 micrograms/kg, nicardipine 10 micrograms/kg and nisoldipine 10 micrograms/kg, decreased aortic blood pressure and increased aortic (AoF), vertebral (VBF) and coronary blood flow (CBF), but did not affect heart rate and left ventricular end diastolic pressure. Renal blood flow was reduced by isradipine 10 micrograms/kg and nifedipine 10 micrograms/kg, but was not influenced by isradipine 3 micrograms/kg, nicardipine 10 micrograms/kg and nisoldipine 10 micrograms/kg. Left ventricular dP/dt was increased by isradipine 3 micrograms/kg, nicardipine 10 micrograms/kg and nisoldipine 10 micrograms/kg, but remained essentially unchanged following isradipine 10 micrograms/kg and nifedipine 10 micrograms/kg. The increase in AoF, VBF and CBF lasted 5-9 min following nifedipine 10 micrograms/kg or nicardipine 10 micrograms/kg, 17-30 min following nifedipine 10 micrograms/kg or nicardipine 10 micrograms/kg, 17-30 min following nisoldipine 10 micrograms/kg, and 16-44 min following isradipine 3 micrograms/kg i.v., but persisted for at least 60 min following isradipine 10 micrograms/kg. Under the experimental conditions and at the doses used in this study, all 4 drugs reduced total peripheral resistance as well as resistance in the vertebral, coronary and renal vascular beds. The results suggest that isradipine exerts cardiovascular effects similar to other calcium antagonists of the dihydropyridine group, but possesses a longer duration of action and shows a greater specificity in reducing coronary vascular resistance than nifedipine, nicardipine and nisoldipine. PMID- 2976589 TI - Application of porous bioceramic in experimental therapy of bone injuries. I. Morphological and histochemical studies in the control animals. AB - The process of healing of the rabbits' bone tissue after mechanical injury was studied. The femoral bone was injured by means of oscillating saw. The animals were killed 3, 6, 9, 14, 18 and 30 days after operation and the callus with the surrounding periosteum was taken for morphological examination. Besides, acid mucopolysaccharides level and aminopeptidase activity in the callus (up to the 18th day of healing) were determined. The time of total bone union was established for 30 days. PMID- 2976591 TI - Maternal mortality at the workers' Hospital, Rangoon (1983-1987) PMID- 2976590 TI - Which is the best test to detect the polycystic ovary? AB - Laparoscopy was used to characterize the ovaries of a group of infertile women. Patients with polycystic ovaries (PCO) were compared with those with normal ovaries. A free androgen index (FAI) greater than 4.5 was 100% specific and sensitive for detecting the PCO in a group of women with oligomenorrhoea. Overall, a biochemical 'screen' consisting of a luteinizing hormone (LH) level greater than 10 U/l, testosterone (T) greater 2.6 nmol/l, sex hormone binding globulin (SHBG) greater than 31 nmol/l or a FAI greater than 4.5 was 89% sensitive and 98% specific for PCO, which is comparable with the reliability of ultrasound for detecting the PCO. PMID- 2976592 TI - Functional significance of ventricular dilatation. Reconsideration of Linzbach's concept of chronic heart failure. AB - On the basis of theoretical considerations and experimental data this study deals with the functional consequences of structural dilatation, particularly in view of Linzbach's concept of chronic heart failure (34-38). After a short review of the literature, a theoretical analysis of the relationship between stroke volume and ventricular inner radius is presented assuming a thick-walled sphere. Presupposing constant contractility, end-diastolic sarcomere length, end diastolic wall thickness and end-systolic pressure, only a considerable increase of ventricular radius could be the direct cause of ventricular pumping failure - despite increasing wall stress and reduced ejection fraction. Impaired contractility, as well as insufficient hypertrophy and increased systemic pressure, would intensify the adverse consequences of ventricular enlargement to a predictable extent. Thus, hemodynamic and energetic consequences of dilation, although mutually interacting, should in principle be distinguished. Despite considerable simplifications involved in model calculations, the relative significance of contractility, ventricular size, wall thickness, and extracardiac factors (mechanical overload; neuroendocrine reactions) can be estimated in various animal models with congestive failure. Hence, this theoretical and experimental approach permits the modification and deepening of previous concepts of structural dilatation and also has implications for interpreting the effects of therapeutical interventions. PMID- 2976593 TI - Effects of physical training on the myocardium of streptozotocin-induced diabetic rats. AB - Effects of endurance swimming training on myocardial contractility and left ventricular myosin isoenzymes were examined in diabetic rats. A diabetic condition was induced in 15-week-old male Wistar rats, by intravenous injection of streptozotocin (50 mg/kg). Swimming training was carried out for five to six weeks (90 min/day, 6 days/week). In order to estimate myocardial contractility, the isometric developed tension of the isolated left ventricular papillary muscle was measured. Myosin isoenzymes were obtained by pyrophosphate gel electrophoresis. Fasting blood glucose of the trained group was significantly lower than that of the sedentary group (sedentary vs. trained = 409.6 +/- 25.9 vs. 266.3 +/- 20.5 mg/dl, p less than 0.001). There was no significant difference in isometric developed tension (T) between the two groups, and the dT/dtmax of the trained group showed a tendency to increase (sedentary vs. trained, T: 2.8 +/ 0.8 vs. 2.9 +/- 0.8 g/mm2, dT/dtmax: 23.1 +/- 3.6 vs. 26.2 +/- 3.5 g/mm2.s, p less than 0.1). Myocardial mechanical responses to isoproterenol and dibutyryl cAMP were increased in the trained group. Left ventricular myosin isoenzyme pattern was shifted towards VM-1 by endurance swimming (sedentary vs. trained, VM 1: 5.6 +/- 4.5 vs. 19.6 +/- 8.8%, p less than 0.001, VM-3: 75.1 +/- 10.0 vs. 54.9 +/- 14.7%, p less than 0.001). These results indicate that endurance swimming can improve disordered glucose metabolism and also influence myocardial contractility, myocardial catecholamine responsiveness, and energetics in myocardial contraction. PMID- 2976595 TI - Tay-Sachs disease: B1 variant. AB - This first child of non-Jewish parents had nystagmus at 4 months of age, bilateral cherry-red macular spots at 7 months of age, and hyperacusis at 8 months of age; the patient has deteriorated progressively following a clinical course typical of Tay-Sachs disease B variant. Total beta-N-acetylhexosaminidase assayed with 4-methylumbelliferyl-beta-glucosamine (4 MU GlcNAc) as substrate was within the normal range in plasma and cultured dermal fibroblasts and 2/3 the normal mean in leukocytes. The hexosaminidase A activity, assayed with the same substrate in plasma and cultured fibroblasts, approximated Tay-Sachs disease heterozygote levels; however, the activity of hexosaminidase A assayed with 4 MU Glc NAc-6-sulfate in the plasma, leukocytes, and cultured fibroblasts was less than 8, 2, and 1%, respectively of the control mean. This female infant with the B1 variant of Tay-Sachs disease demonstrated an earlier onset and more rapidly progressive course than was observed in 4 of the 5 previously reported patients with this Tay-Sachs disease variant. PMID- 2976596 TI - [Epidemiology of cancer of the uterine cervix in Costa Rica, 1980-1983]. PMID- 2976594 TI - Influence of spontaneous hypertension and cardiac hypertrophy on the severity of ischemic arrhythmias in the rat. AB - Cardiac hypertrophy (CH) and hypertension (HT) are major determinants of sudden cardiac death in patients with coronary artery disease. To investigate the hypothesis that CH and HT increase the incidence of severe ventricular arrhythmias in an animal model, we performed a 30-min period of coronary artery ligation in anesthetized spontaneously hypertensive rats (SHR), normotensive Wistar Kyoto (WKY) and Wistar (W) rats. The incidence and duration of ventricular fibrillation resulting from coronary artery occlusion were significantly (p less than 0.01) increased in hypertensive rats compared to normotensive animals. The calcium entry blocker nicardipine was administered orally to SHR either chronically for 8 weeks (20 mg.kg-1 twice daily) or acutely as a single dose of 20 mg.kg-1. After long-term treatment with nicardipine, left ventricular hypertrophy index and systolic blood pressure were significantly (p less than 0.001) reduced when compared to vehicle-treated SHR, whereas a single administration of nicardipine only decreased blood pressure without affecting cardiac mass. In the long-term nicardipine-treated SHR group, acute coronary artery ligation induced significantly less ventricular fibrillation (p less than 0.05) and mortality (p less than 0.001) than in acutely nicardipine-treated or untreated SHR groups. In conclusion, the data suggest that the severity and incidence of lethal ventricular arrhythmias are more elevated in hypertensive than in normotensive rats and this may be related to the myocardial hypertrophic state. PMID- 2976597 TI - [Cancer of the stomach in Costa Rica. Incidence and mortality]. PMID- 2976598 TI - [The demand for the main cardiovascular preparations in the city of Frunze]. AB - The structure of consumption and need in cardiovascular drugs were studied in a hospital and in the Kirghiz Scientific Research Institute of Cardiology according to a scheme developed in the USSR Cardiology Research Center. Use of cardiovascular drugs was studied with respect to optimal results of therapy in 361 patients with main cardiac pathology (ischemic heart disease, essential hypertension and their combination) who had received 1125 courses of drug treatment. The prescriptions were analysed according to their intensity, regularity and optimal effect. The authors assessed provision of cardiological patients of Frunze with drugs and gave recommendations on its improvement, in particular, satisfaction of need in some active drugs (prolonged nitrates, calcium antagonists, cardiac glycosides). PMID- 2976599 TI - Integration of Down's syndrome children in the primary school: a longitudinal study of cognitive development and academic attainments. PMID- 2976600 TI - On the intermediacy of carboxyphosphate in biotin-dependent carboxylations. AB - In the ATP-dependent carboxylation of biotin that is catalyzed by most biotin dependent carboxylases, a fundamental mechanistic question is whether the ATP activates bicarbonate (via the formation of carboxyphosphate as an intermediate) or whether the ATP activates biotin (via the formation of O-phosphobiotin). We have resorted to three mechanistic tests using the biotin carboxylase subunit of acetyl-CoA carboxylase from Escherichia coli: positional isotope exchange, intermediate trapping, and 18O tracer experiments on the ATPase activity. First, no catalysis of positional isotope exchange in adenosine 5'-[( alpha, beta-18O, beta, beta-18O2]triphosphate) was observed when either biotin or bicarbonate was absent, nor was any exchange seen in the presence of both N-1-methylbiotin and bicarbonate. Second, the putative carboxyphosphate intermediate could not be trapped as its trimethyl ester, under conditions of incubation and analysis where the authentic triester was shown to be adequately stable. In the third test, however, we showed that the ATPase activity of biotin carboxylase that is seen in the absence of biotin, an activity that is known to parallel the normal carboxylase reaction when biotin is present, occurs with the transfer of an 18O label directly from [18O]bicarbonate into the product Pi. This result suggests that the bicarbonate-dependent biotin-independent ATPase reaction catalyzed by biotin carboxylase goes via carboxyphosphate and that the carboxylation of biotin itself may proceed analogously. PMID- 2976601 TI - Sites in the diyne-ene bicyclic core of neocarzinostatin chromophore responsible for hydrogen abstraction from DNA. AB - The antitumor antibiotic neocarzinostatin exhibits its main drug action by abstracting hydrogen from DNA deoxyribose with consequent strand breakage or related lesions. All biological activities of the drug derive solely from a nonprotein chromophoric substance (NCS-chrom) consisting of a novel epoxy-bicyclo diyne-ene system. Thiol or sodium borohydride activates NCS-chrom into a labile, reactive species that induces DNA damage but causes inactivation of the drug in the absence of the target DNA. Mass spectrometric studies indicate that the isolated thiol-activated NCS-chrom product in the presence of DNA has the same molecular weight as the thiol-inactivated NCS-chrom product in the absence of DNA. No deuterium is incorporated into the chromophore from the deuterium-labeled sulfhydryl group. Since three deuterium atoms can be incorporated into the drug by treatment with sodium borodeuteride without DNA, adding an unlabeled DNA under parallel conditions permitted the ready identification of the activated NCS-chrom product that abstracted hydrogen from the DNA. Not only does the activated NCS chrom product have the same structure as the inactivated drug without DNA, but two of the incorporated deuterium atoms have been substituted by hydrogen. With the aid of NMR spectrometry, the two replaced hydrogen atoms are found to be incorporated into the C-2 and C-6 positions of the bicyclo-diyne-ene ring of NCS chrom and are derived neither from borodeuteride nor from the hydroxyl functions of the solvents. In accord with current proposals, the two hydrogens incorporated into the drug may come from closely opposed sites on the complementary strands of the DNA at which the drug is bound.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976602 TI - Docosahexaenoate-containing molecular species of glycerophospholipids from frog retinal rod outer segments show different rates of biosynthesis and turnover. AB - We have studied the de novo synthesis and subsequent turnover of major docosahexaenoate-containing molecular species in frog rod outer segment (ROS) phospholipids following intravitreal injection of [2-3H]glycerol. On selected days after injection, ROS were prepared and phospholipids extracted. Phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS) were isolated and converted to diradylglycerols with phospholipase C. Diradylglycerols were derivatized with benzoic anhydride and resolved into diacylglycerobenzoates and ether-linked glycerobenzoates. The diacylglycerobenzoates were fractionated into molecular species by HPLC, quantitated, and counted for radioactivity. Label was incorporated into ROS phospholipids by day 1 and was followed up through the eighth day. The dipolyenoic species 22:6-22:6 from PC showed a 3-5 times higher radiospecific activity than the same species from either PE or PS. In PC, the specific activities of 16:0-22:6 and 18:0-22:6 were 3-5 times lower than the specific activity of 22:6-22:6. In contrast, for PE, the specific activities of 16:0-22:6 and 18:0-22:6 were 2-5 times higher than that of 22:6-22:6. The specific activities of 18:0-22:6 and 22:6-22:6 were similar in PS. Specific activities of the docosahexaenoate-containing species began approximating an exponential decline 6-8 days postinjection and continued through the 31st day. The rate of decline was determined by calculating the half-life of each molecular species, which was used as a measure of the turnover of the species. The species 22:6-22:6 PE and 18:0-22:6-PE showed a 2-3 times slower turnover rate than the corresponding species from either PC or PS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976603 TI - [The effect of proteolysis of myosin heads on conformational changes induced by them in F-actin]. AB - The effect of limited proteolysis of myosin subfragment I (S1) on conformational changes in F-actin during the formation of a rigor F-actin-S1 complex was studied, using polarized microfluorimetry. Upon the decoration of thin filaments made up of rhodaminyl-phalloin modified F-actin with subfragment I, the anisotropy of fluorescence increased. Limited proteolysis of S1 at the junctions of 27 kD-70 kD and 27 kD-50 kD-20 kD fragments as well as the destruction of the 50 kD fragment by methanol had no effect on the nature of these changes. It was assumed that during the formation of the actomyosin complex the conformational changes in F-actin were induced by the 20 kD fragment. The 27 kD and 50 kD fragments seemed to exert a weak influence on action conformation within the complex. PMID- 2976604 TI - The non immunogenicity of CALLA in leukemic children. AB - The sera from 55 children with acute lymphoblastic leukemia (ALL) treated with active immunotherapy were examined for the presence of antibodies against common ALL antigen (CALLA). A solid phase indirect radioimmunometric assay (IRA) was developed which enabled the detection of anti-CALLA antibody in patients' sera, utilizing the ability of affinity-purified CALLA to bind Ricinus communis agglutinin and anti-CALLA antibody simultaneously. Using IRA, anti-CALLA antibody activity could not be detected in a majority of patients. We concluded that the patients did not raise comparable antibodies against CALLA, indicating this antigen is not immunogenic for ALL patients. PMID- 2976606 TI - Aortic angioplasty. AB - Percutaneous transluminal angioplasty is a well-recognized and accepted alternative to surgery in the treatment of arterial stenoses in the peripheral vasculature, coronary arteries and aortic branches such as the renal arteries. Its application in the abdominal aorta is less well described but in selected patients and with experienced operators it has been shown to be effective. PMID- 2976605 TI - Hexachlorophene distributions in estuarine sediments. PMID- 2976607 TI - Incisions with knife or diathermy and postoperative pain. PMID- 2976609 TI - Rectus abdominis muscle flap for persistent perineal sinus. PMID- 2976608 TI - A protocol for the safe treatment of acute lower limb ischaemia with intra arterial streptokinase and surgery. AB - Over a 5-year period 70 patients, presenting with subacute ischaemia of the lower limb (more than 12 h), were treated with low-dose intra-arterial streptokinase. There were 72 infusions and effective lysis was achieved in 52 (72 per cent), with an average infusion time of 25 h. A total of 23 (32 per cent) also underwent percutaneous transluminal angioplasty when lysis showed an underlying stenosis, and a further 19 (26 per cent) required surgery to remove persistent stenosis, organized thrombus or atheromatous debris. Significant bleeding occurred in 4 patients (6 per cent) and 13 (18 per cent) underwent amputation. There were five deaths (7 per cent), one of which was directly related to the infusion, while three were due to myocardial infarction. All of the major complications occurred in the early part of the study and both the selection of patients and the technique of infusion were modified to improve safety. Complementary treatment by percutaneous transluminal angioplasty and surgery was used more frequently in the later part of the study. The technique is not recommended for the white leg of acute ischaemia (less than 12 h), or for lysis of clot in a retroperitoneal Dacron graft, but may be uniquely valuable to demonstrate the cause of subacute ischaemia. PMID- 2976610 TI - [Normal school and medicosocial integration of young handicapped subjects]. PMID- 2976611 TI - Phage typing set for differentiating Staphylococcus epidermidis. AB - A phage typing set composed of 13 phages is described for characterizing Staphylococcus epidermidis. Isolates (372) from cases of bovine mastitis were used in this study. Of these, 350 or 94% were successfully delineated, and 63 phage types were observed. Twenty two cultures were not typeable. PMID- 2976612 TI - [Diagnosis of left ventricular hypertrophy in arterial hypertension: comparison of echocardiography and electrocardiography]. PMID- 2976613 TI - [Electrocardiographic, vectorcardiographic and echocardiographic study of the grade of left ventricular hypertrophy in trained and semi-trained athletes]. PMID- 2976614 TI - Pinealocytes immunoreactive with antisera against secretory glycoproteins of the subcommissural organ: a comparative study. AB - By means of light-microscopic immunocytochemistry two polyclonal antibodies (AFRU, ASO; see p. 470) directed against secretory glycoproteins of the subcommissural organ were shown to cross-react with cells in the pineal organ of lamprey larvae, coho salmon, a toad, two species of lizards, domestic fowl, albino rat and bovine (taxonomic details, see below). The AFRU-immunoreactive cells were identified as pinealocytes of the receptor line (pineal photoreceptors, modified photoreceptors or classical pinealocytes, respectively) either due to their characteristic structural features or by combining AFRU immunoreaction with S-antigen and opsin immunocytochemistry in the same or adjacent sections. Depending on the species, AFRU- or ASO-immunoreactions were found in the entire perikaryon, inner segments, perinuclear area, and in basal processes facing capillaries or the basal lamina. In most cases, only certain populations of pinealocytes were immunolabeled; these cells were arranged in a peculiar topographical pattern. In lamprey larvae, immunoreactive pinealocytes were observed only in the pineal organ, but not in the parapineal organ. In coho salmon, the immunoreaction occurred in S-antigen-positive pinealocytes of the pineal end-vesicle, but was absent from S-antigen-immunoreactive pinealocytes of the stalk region. In the rat, AFRU-immunoreaction was restricted to S-antigen immunoreactive pinealocytes found in the deep portion of the pineal organ and the habenular region. These findings support the concept that several types of pinealocytes exist, which differ in their molecular, biochemical and functional features. They also indicate the possibility that the AFRU- and ASO immunoreactive material found in certain pinealocytes might represent a proteinaceous or peptide compound, which is synthesized and released from a specialized type of pinealocyte in a hormone-like fashion. This cell type may share functional characteristics with peptidergic neurons or paraneurons. PMID- 2976615 TI - Circadian rhythm of prohormone atrial natriuretic peptides 1-30, 31-67 and 99-126 in man. AB - Two peptides consisting of amino acids 1-30 and 31-67 of the N-terminal end of the prohormone of the atrial natriuretic factor (pro ANF), vasodilate aortas in vitro, lower blood pressure in vivo, and have natriuretic properties similar to the atrial natriuretic factor (ANF, amino acids 99-126 of the prohormone). It has been recently discovered that pro ANF 1-30 and pro ANF 31-67 as well as ANF circulate in man. To determine if these three peptide hormones have a circadian variation in their circulating plasma concentrations, eight housestaff volunteers were studied on a day when they were in the hospital for 24 hr. These 5 men and 3 women, ages 25 to 39 had blood samples taken at 0800, 1200, 1600, 2000, 0000, 0400 and 0800 on the following day. One-half of these house officers were up all night while the other half went to sleep from midnight to 0800 and had their 0400 plasma samples drawn while in a supine position. The peak level for all three peptide hormones was at 0400 for both supine and upright subjects. It was concluded that there are circadian rhythms in normal, active people of these three peptide hormones, whose peak levels are at 0400 irrespective of posture. PMID- 2976616 TI - Ovarian abscess following cesarean section. A case report and review of literature. AB - Ovarian abscess is presented although such a finding is an unusual gynecologic complication. It is difficult to distinguish from a tubo-ovarian abscess. Its presence may be suspected in a patients after surgery, carrying IUD, having intraperitoneal infection and pregnancy. If an abscess is present, a tubo-ovarian abscess is much more common, except in pregnancy. For the last 110 years only 120 cases of ovarian abscesses have been reported in the Literature. PMID- 2976617 TI - Hormonal profile and androgen status during the menstrual cycle in women with acne. AB - The androgen excess in women with acne can play an important role in gynecological endocrinology. LH, FSH, PRL, Testosterone (T), Androstenedione (A), Deydroepiandrosterone sulphate (DHEA-S) and Progesterone (P) were investigated in 21 women with uncomplicated idiopathic acne during the menstrual cycle. LF/FSH ratio greater than 3 was found in 7 cases (36.8%). Total T values were high in 90.4% of the whole group of women with acne (mean +/- SD 618.6 +/- 253.6, normal range 92-352 pg/ml). An ovulatory pattern of P was found in 66.65% of women with acne. Hyperprolactinemia was found in 23.6% of cases. The common denominator for ovulatory dysfunction in these women seems to be elevated LH levels, rather than elevated T levels. PMID- 2976618 TI - Correlation between concentrations of magnesium, zinc, and potassium in plasma, erythrocytes and muscles. AB - In 93 patients, consecutively subjected to laparotomy, we studied the correlation between concentrations of magnesium, zinc and potassium in plasma, erythrocytes and muscle tissue. Sixteen percent had hypomagnesemia, 27% hypozincemia, and 53% hypokalemia. There was no difference with regard to the concentration of magnesium and zinc in erythrocytes or in muscle tissue between patients with low and patients with normal/high plasma concentrations of these minerals. Potassium concentration per liter erythrocytes was slightly but significantly higher in patients with hypokalemia than in patients with normal plasma potassium. When expressed per mmol haemoglobin there was, however, no difference in erythrocyte potassium between these two groups. Neither did they differ with regard to potassium concentration in muscles. There was a highly significant correlation between the concentrations of magnesium, zinc and potassium in erythrocytes as well as in muscle tissue, and a significant correlation between magnesium and potassium in plasma. However, no correlation was found between the mineral concentrations in plasma, erythrocytes and muscles, neither for magnesium, nor for zinc or potassium. We therefore conclude that a valid test to estimate total body stores of these minerals cannot be based on analysis of erythrocytes. PMID- 2976620 TI - Induction of the in vitro anti-HBs response by hepatitis B surface antigen. AB - The production in vitro of antibody to hepatitis B surface antigen by peripheral blood mononuclear cells of healthy volunteers was studied after each of the three doses of hepatitis B vaccine. An in vitro hepatitis B surface antigen antibody response was successfully induced in 12% of the specimens taken over a 7 month period. The response to this antigen was induced in additional samples if cells had been treated previously with anti-CD4 and complement or anti-CD8 and complement prior to culture initiation. The addition of interleukin 2 could also induce the formation of antibodies to hepatitis B surface antigen. The results suggest that the antibody response to hepatitis B surface antigen is complex and varies depending on the individual and time of sampling. PMID- 2976619 TI - Immunologic abnormalities related to antigenaemia during HIV-1 infection. AB - The expression of phenotypic markers on T and B lymphocytes in long-term human immunodeficiency virus type 1 (HIV-1) seropositive, antigen negative patients, in seropositive, antigen positive individuals without AIDS and in seronegative intravenous drug abusers was examined by two colour flow cytometry. Seropositive, antigen positive patients showed decreased CD4+ lymphocyte numbers, causing lower CD4/CD8 ratios when compared to seropositive, antigen negative subjects. While CD4 CDw29+ (4B4) lymphocytes are selectively reduced in seropositive, antigen negative individuals, both CD4 CDw29+ and CD4 CD45R+ (2H4) lymphocytes are decreased when antigenaemia is present. An increased percentage of CD3 HLA DR+ activated T lymphocytes and of CD20+ (B1) Leu 8 negative activated B cells was seen in HIV-1 seropositive antigen positive patients. These results demonstrate that, in long-term seropositive individuals, antigenaemia is associated with peculiar phenotypic changes of lymphocyte subsets. PMID- 2976621 TI - Nickel antigen induces IL-2 secretion and IL-2 receptor expression mainly on CD4+ T cells, but no measurable gamma interferon secretion in peripheral blood mononuclear cell cultures in delayed type hypersensitivity to nickel. AB - Nickel sulphate antigen-induced peripheral blood lymphocyte activation in vitro was characterized by lymphokine measurement (IL-2, IFN-gamma) and phenotyping of the IL-2 responsive cells. Mononuclear cells from nickel-sensitive patients synthesized more DNA, produced more IL-2 and had more IL-2 receptor positive cells in response to nickel than did those of the control subjects. On the other hand no IFN-gamma was detectable in the nickel supernatants, while PPD, used as the control antigen, induced pronounced quantities of IFN-gamma with an equal amount of DNA synthesis. The increase in IL-2 receptor positive cells was due to activation of CD4+ (helper/inducer) T cells. T cells with HLA-DR antigen surface markers were more numerous on each day of culture than cells with IL-2 receptors. These two activation markers were co-expressed on the same cells only to a certain extent, thus perhaps reflecting different types or phases of activation. In conclusion, nickel-induced peripheral blood mononuclear cell activation in vitro differs from microbial antigen-induced activation with respect to its modest or non-existent IFN-gamma response. PMID- 2976622 TI - Role of the spleen in immunosuppression of gastric cancer: predominance of suppressor precursor and suppressor inducer T cells in the recirculating spleen cells. AB - In order to analyse the role of the spleen on immunosuppression of gastric cancer, T cell phenotypes in the spleen cells (SC) were investigated by two colour fluorescence flow cytometry, with reference to their suppressor cell activity. Suppressor T cell phenotypes of CD4+2H4+ cells (suppressor/inducer T cells) and CD8+CD11+ (suppressor T cells) were distributed predominantly in SCs from patients with gastric cancer, while they were distributed scarcely in those with liver cirrhosis. Moreover, CD4+2H4+ cells and CD8+CD11+ cells were found predominantly in SCs and splenic vein lymphocytes (SVL) respectively. Among SCs, a significantly higher proportion of CD4+2H4+ cells was found in the recirculating SCs, but fewer were found in the residual SCs. Higher activity of Concanavalin-A induced suppressor cells was found in the former and that of spontaneously activated suppressor cells was found in the latter. These results suggest the suppressor precursor and suppressor/inducer T cells might distribute predominantly in the cells recirculating from the spleen, and that suppressor cells might be matured during the migration from the spleen. PMID- 2976623 TI - Cyclosporin A induces a selective, reversible suppression of T-helper lymphocyte regeneration after syngeneic bone marrow transplantation: association with syngeneic graft-versus-host disease in rats. AB - Experimental studies in rats have established that a combination of lethal X irradiation, syngeneic bone marrow transplantation (sBMT) and temporary administration of Cyclosporin A (CyA) leads to the development of a disease defined as syngeneic graft-versus-host disease or CyA-induced autoimmunity. This study demonstrates that the combination of lethal X-irradiation, sBMT, and CyA selectively suppresses the repopulation of T-helper cells, as monitored in peripheral blood. The suppression of T-helper cell regeneration is maintained as long as CyA is administered. In addition, development of CyA-induced autoimmunity does not start as long as CyA is given. After withdrawal of CyA T-helper cells recover. Their reappearance in the peripheral circulation coincides with the development of syngeneic graft-versus-host disease, suggesting a role of these cells in initiating or causing the disease. PMID- 2976624 TI - Cost and efficacy of intravenous streptokinase plus PTCA for acute myocardial infarction when therapy is initiated in community hospitals. AB - The costs and clinical results of a thrombolytic therapy protocol using intravenous (IV) streptokinase in small community hospitals prior to emergency transfer for cardiac catheterization for possible percutaneous transluminal coronary angioplasty (PTCA) were prospectively studied in 56 patients with an acute myocardial infarction [Tulsa Heart Center-Myocardial Infarction Study Team (THC-MIST) protocol]. Findings were compared to those for 53 similar patients receiving "conventional therapy" during the same study period. The reperfusion rate for IV streptokinase was 63%. Additional emergency PTCA yielded a total reperfusion rate of 95%. Despite emergency cardiac catheterization for all THC MIST patients, average hospital charges were not significantly different between the two groups ($20,495 vs. $20,722; THC-MIST vs. conventional therapy, respectively). Total charges (hospital plus physician fees) were also not significantly different ($22,986 vs. $22,400; THC-MIST vs. conventional therapy, respectively). Intensive care unit (ICU) days and total hospital stays were significantly less for THC-MIST patients (3.1 vs. 5.8 ICU days, p less than 0.05 and 8.8 vs. 12.4 hospital days, p less than 0.05; THC-MIST vs. conventional therapy, respectively). Thus initiation of IV streptokinase in community hospitals for acute myocardial infarction with subsequent transfer for possible PTCA can yield high reperfusion rates without significantly increasing health care costs. PMID- 2976626 TI - Morphologic correlates of coronary angiographic patterns at the site of percutaneous transluminal coronary angioplasty. AB - Over the last 10 years considerable interest has been paid in the angiographic recognition of successful percutaneous transluminal coronary artery angioplasty (PTCA), complication of the technique, and angiographic predictors of restenosis. This report summarizes various angiographic patterns visualized at the site of angioplasty and correlates these patterns with morphologic findings. Of 66 patients undergoing PTCA for chronic or unstable angina pectoris, 76 PTCA sites were available for analysis. The two most common angiographic patterns at the angioplasty site (intimal flap = 43%, intraluminal haziness = 38%) correlated morphologically with intimal-medial splits with localized dissections (79%). Shallow, superficial intimal lesions, laminated thrombus, and adventitial tears accounted for the remaining morphologic changes. Eight PTCA sites without morphologic injury corresponded to smooth wall changes and spasm at angiography. PMID- 2976625 TI - Changes in the levels of plasma atrial natriuretic peptide, hemodynamic measurements, and the levels of vasoactive hormones during the clinical course of congestive heart failure. AB - To investigate the mechanism for the release of human atrial natriuretic peptide (hANP) and the pathophysiological role of hANP in patients with congestive heart failure (CHF), plasma hANP levels in patients with dilated cardiomyopathy (DCM) or acute myocardial infarction (AMI) were determined serially, and the relationship between plasma ANP levels and hemodynamic measurements or various vasoactive hormones was analyzed during the clinical course of congestive heart failure. In 63 patients with either AMI or DCM, plasma hANP, plasma renin activity, aldosterone concentration, and catecholamines were measured over 4 weeks, during the course of CHF. Cardiac catheterization with a Swan-Ganz catheter was also performed. Plasma hANP in patients with DCM was elevated continuously during the clinical course. Plasma hANP levels in patients with AMI of Groups II and IV of Forrester's class decreased on days 7 and 14 and those in patients with AMI of Group I changed within normal limits. Plasma hANP levels were correlated positively with pulmonary artery pressure and pulmonary capillary wedge pressure in patients with AMI or DCM. Plasma renin activity, noradrenaline, and adrenaline levels were elevated in the acute phase of myocardial infarction and had a tendency to decrease upon improvement in clinical status. Plasma renin activity and noradrenaline level correlated positively with plasma hANP levels. These data indicate that plasma hANP levels are regulated by atrial distension and severity of cardiac impairment, and that plasma hANP and plasma renin activity or catecholamines correlated closely during the clinical course of CHF, indicating that these hormones may be involved in the volume and electrolytes status in CHF. PMID- 2976627 TI - Anti-androgens in gynaecological practice. AB - Hirsutism and acne in women are common distressing problems. Unwanted hair growth, acne and seborrhoea result from the action of androgens on the skin. Such effects depend not only on increased androgen production by the ovary or adrenal gland but also on the bioavailability of androgen to peripheral tissues. This in turn is related to transport of androgens in plasma by specific binding proteins and to peripheral metabolism of testosterone and androstenedione to their more potent 5 alpha-reduced derivatives. An effective anti-androgen is one which blocks the androgen receptor-mediated actions of testosterone and DHT on skin. CPA, the treatment of choice in the UK, is a potent androgen receptor-blocking steroid which also has progestational properties. When combined with ethinyloestradiol it also suppresses ovarian function, thus reducing androgen production, and provides effective contraception. PMID- 2976628 TI - Tumors of the cauda equina: the importance of an early diagnosis. AB - The natural history of cauda equina tumors was studied in 13 patients. Pain was the most prominent clinical manifestation. Neurological dysfunction mainly consisted of sensory disturbances. The pre-operative course was in 62% three years or longer. A relation was found between the histology of the tumor and the delay of diagnosis: ependymomas were discovered much later than schwannomas. Other histological types of tumors were meningioma, sarcoma and metastasis of a medulloblastoma. The effect of treatment, in nine cases only surgical, was excellent in seven of the 13 patients. The prognosis was determined by histology and length of pre-operative period. PMID- 2976629 TI - Spontaneous extrusion of peritoneal catheter through intact abdominal wall. AB - A rare occurrence of spontaneous extrusion of a Denver peritoneal catheter through the right flank in a 14-year-old boy is reported. The firm texture and short length of the catheter coupled with its movements with respiration produced a hammer effect and eroded the abdominal wall. Disconnection of the peritoneal catheter from the chamber and pulling out the extruded catheter is suggested as a simple and effective method of removal of the shunt. PMID- 2976630 TI - Quantitative analysis of a new marker for common acute lymphoblastic leukemia detected by two-dimensional electrophoresis. AB - A major subtype of childhood lymphoblastic leukemia has been previously defined on the basis of expression of a cell surface marker referred to as the common acute lymphoblastic leukemia antigen. In this study we provide evidence that leukemic cells from most of the patients with common acute lymphoblastic leukemia contain a cytosolic polypeptide designated L4, the occurrence of which is strongly correlated with the expression of the common acute lymphoblastic leukemia antigen. The detection and quantitation of L4 was accomplished by analysis of cellular polypeptides resolved by two-dimensional polyacrylamide gel electrophoresis. This approach provides a powerful tool for the delineation of distinct polypeptides corresponding to different types of leukemia and complements immunological analysis of the cell surface marker phenotype. PMID- 2976632 TI - [Spontaneous reporting. Significance, applications and interpretation of the data]. PMID- 2976631 TI - [Treatment of arterial hypertension for cardio-cerebral protection: role of calcium antagonists]. PMID- 2976633 TI - [Evaluation of the efficacy of profibrinolytic treatment with defibrotide in the therapy of thrombophlebitis]. PMID- 2976634 TI - [Functional dyspeptic syndrome caused by dyskinesia of the bile ducts. Auscultatory percussion diagnosis and therapeutic monitoring]. PMID- 2976635 TI - [Use of prosthesis for vascular access in hemodialysis]. PMID- 2976636 TI - [Gastroenteropancreatic endocrine system and crenotherapy with mineral waters]. PMID- 2976637 TI - [Magnetic resonance of the adrenal glands]. PMID- 2976638 TI - [Magnetic resonance of the head and neck in pediatrics]. PMID- 2976639 TI - Promoting better health: the missing strategy. PMID- 2976640 TI - Diabetes in the United Kingdom: some facts and figures. PMID- 2976641 TI - Questionnaire design in diabetes care and research. 2: Making the choice. PMID- 2976642 TI - Association of hypertension with blood viscosity in diabetes. AB - Plasma and whole blood viscosity and its determinants were measured in 86 diabetic patients (29 hypertensive and 57 normotensive) and compared with 52 non diabetic control subjects to assess whether hypertension has an additive and adverse effect on blood viscosity. Whole blood viscosity (corrected for haematocrit), at high and low shear rates (95 and 0.95 s-1), was significantly higher in both Type 1 (5.1 +/- 0.5 (+/- SD), 19.8 +/- 2.9) and Type 2 (5.2 +/- 0.3, 21.1 +/- 2.0) diabetic patients compared with control subjects (4.9 +/- 0.6, 17.4 +/- 2.6 mPa s, p less than 0.01). Corrected whole blood viscosity at high shear rate was significantly higher in hypertensive than in normotensive Type 2 diabetic patients (5.5 +/- 0.4 vs 5.2 +/- 0.3 mPa s, p less than 0.01). Plasma viscosity was significantly higher in diabetic patients compared with control subjects (1.4 +/- 0.1 vs 1.3 +/- 0.1 mPa s, p less than 0.01), but there was no difference between hypertensive and normotensive diabetic patients (1.4 +/- 0.1 vs 1.4 +/- 0.2 mPa s). Fibrinogen levels were similar in all the groups. PMID- 2976643 TI - Case-control study on risk factors associated with fibrocalculous pancreatic diabetes. AB - We investigated the relation between fibrocalculous pancreatic diabetes and cassava consumption in a case-control study, in which 31 cases of pancreatic diabetes were compared with 45 non-diabetic control subjects who had no pancreatic calcification. Risk of diabetes was not related to cassava consumption. We also observed no increased risk of fibrocalculous pancreatic diabetes associated with alcohol consumption, history of gallbladder and biliary tract diseases, ascariasis, and family history of diabetes. Lower monthly income, farmer occupation, rural residence, and low BMI were significantly (p less than 0.05) related to pancreatic diabetes. Our data suggest that consumption of cassava may not be an important risk factor for pancreatic diabetes. With limited sample size, however, cassava consumption could not be excluded as one possible cause of fibrocalculous pancreatic diabetes. PMID- 2976644 TI - Cardiac hypertrophy in diabetic nephropathy: an echocardiographic study. AB - Echocardiography was used to study the prevalence and severity of left ventricular hypertrophy in patients with established diabetic nephropathy (persistent proteinuria for at least 2 y plus severe retinopathy). Fifteen patients had mild renal impairment (serum creatinine less than 150 mumol l-1), 14 patients had moderate renal impairment (serum creatinine 150-400 mumol l-1), and 20 patients had severe renal impairment (serum creatinine greater than 400 mumol l-1). Thirty-six of the 49 (73%) were on anti-hypertensive treatment, despite which mean blood pressure was 161 +/- 25/89 +/- 9 (+/- SD) mmHg. Left ventricular hypertrophy was demonstrated in 42 of the 49 patients (85%), and increased in severity with increasing renal impairment. Interventricular septal + left ventricular posterior wall thickness was 25 +/- 3 mm in those with mild renal impairment, 28 +/- 6 mm in those with moderate renal impairment and 30 +/- 4 mm in those with severe renal impairment. The most severe left ventricular hypertrophy was seen in the Afro-Caribbean patients. Left ventricular hypertrophy was present even in those with marginally raised blood pressure and was related to age and serum creatinine but not to present blood pressure or duration of proteinuria. PMID- 2976645 TI - Cell-mediated immunity and symptomatic diabetic autonomic neuropathy. AB - T lymphocytes have been implicated in the nerve damage observed in allergic experimental neuritis and in idiopathic polyneuritis. Symptomatic autonomic neuropathy in long-standing Type 1 diabetes is a rare and unexplained complication, and some preliminary evidence has suggested a pathogenetic role for the immune system. We have measured levels of activated T lymphocytes in 18 Type 1 diabetic patients with symptomatic autonomic neuropathy and in 16 matched patients with uncomplicated Type 1 diabetes. Purified T lymphocytes from peripheral blood were stained with a fluorescein-labelled monoclonal antibody directed to the activation marker HLA-DR and counted under UV microscopy. Percent DR positive T lymphocytes were significantly raised in the patients with autonomic neuropathy when compared with long-standing uncomplicated diabetic patients (8.2 +/- 4.2 vs 4.9 +/- 3.3%, p less than 0.01). This finding lends support for a role of the immune system in the development of autonomic neuropathy. PMID- 2976646 TI - Blood glucose control and insulin secretion improved with combined therapy in type 2 diabetic patients with secondary failure to oral hypoglycaemic agents. AB - The influence of combined therapy using insulin and oral hypoglycaemic agents on blood glucose control and on insulin secretion in Type 2 diabetic patients with secondary failure to oral hypoglycaemic agents was evaluated. Type 2 diabetic patients (n = 180) (98 normal-weight, 82 over-weight), at least 3 years from diagnosis, and having poor blood glucose control on oral hypoglycaemic agents for at least 3 months (fasting plasma glucose greater than 10.0 mmol l-1) despite intensive efforts at improvement, were included in the study. A single daily insulin injection (human ultralente), at a dose of 0.22 +/- 0.07 U kg-1 d-1 in normal-weight and 0.33 +/- 0.10 U kg-1 d-1 in over-weight patients, was added to the previous dietary and drug treatment for 6 months. A progressive and significant (2p less than 0.001) reduction of the mean daily blood glucose was observed during the first 3 months of combined therapy (from 13.2 +/- 3.2 to 8.1 +/- 2.1 mmol l-1 in normal-weight and from 13.4 +/- 3.1 to 8.8 +/- 2.3 mmol l-1 in over-weight patients), with no further significant changes thereafter. A significant increase (2p less than 0.001) in the mean daily C-peptide concentration (from 0.50 +/- 0.30 to 0.71 +/- 0.29 nmol l-1 in normal-weight and from 0.78 +/- 0.36 to 1.00 +/- 0.41 nmol l-1 in over-weight patients) took place during combined therapy. No changes of body weight (+ 1.5 +/- 1.2 kg in normal weight and + 1.0 +/- 1.0 kg in over-weight patients) were observed. PMID- 2976647 TI - Suppression of insulin secretion by falling plasma glucose levels is impaired in type 2 diabetes. AB - The ability of Type 2 diabetic patients to suppress islet B-cell secretion in response to falling plasma glucose levels has been studied with two different protocols. (1) Five diet-treated diabetic patients and 6 normal subjects were studied after the termination of a hyperglycaemic clamp at 15 mmol l-1 for 150 min, with the plasma glucose levels then being allowed to fall and the glucose clamp re-established at 10 mmol l-1. The plasma insulin levels fell in normal subjects from 178 +/- 141 (+/- SD) mU l-1 at the end of the 15 mmol l-1 clamp to 147 +/- 97 mU l-1 (p less than 0.02) 20 min later, whereas in diabetic patients there was no significant change from 61 +/- 41 to 56 +/- 35 mU l-1, respectively (NS). (2) The second study was performed to assess the turn-off of islet B-cell secretion with diabetic patients and normal subjects starting at comparable plasma insulin levels. Twelve diet-treated diabetic patients and 11 normal subjects were given a continuous low-dose glucose infusion for 60 min at a rate of 5 mg kg-1 ideal body weight min-1, after which the infusion was turned off and the plasma glucose level allowed to fall.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976648 TI - Altered hand skin blood flow in type 1 (insulin-dependent) diabetes mellitus. AB - Disturbed upper limb skin blood flow has been described in insulin-dependent (Type 1) diabetes mellitus, but the pathophysiological mechanism remains unclear. Hand skin blood flow was therefore measured at room temperature and following immersion of hands in cold and warm water in 13 healthy control subjects, in 10 patients with Type 1 diabetes mellitus and cardiovascular autonomic neuropathy, and a further 10 Type 1 diabetic patients with normal cardiovascular autonomic tone. Following cold challenge there was failure of digital artery clampdown in all diabetic patients in comparison with healthy control subjects (p less than 0.005), and the index finger temperature fell less (p less than 0.05). Laser Doppler flow was reduced at the palms at room temperature or following the warm challenge (p less than 0.008), as well as on the dorsum at room temperature (p less than 0.05), in all diabetic patients. In addition laser Doppler flow in the diabetic patients was reduced at the palms and dorsum immediately following cold water challenge (p less than 0.004) and this reduction persisted 15 min (p less than 0.05) and 30 min (p less than 0.01) into the recovery phase. In comparison to those diabetic patients with normal cardiovascular tone, those with cardiovascular autonomic neuropathy had reduced laser Doppler flow at the pulp 15 min after cold water immersion (p less than 0.05), at the nailbed immediately after cold water immersion (p less than 0.01), and at the palms immediately after warm water challenge (p less than 0.01). PMID- 2976649 TI - Acute changes in blood glucose concentration do not promote thrombin generation or fibrin breakdown in type 1 diabetes. AB - To investigate the effect of blood glucose concentration on thrombin generation and fibrinolytic activity, six Type 1 patients had the blood glucose concentration maintained for 1 h at 5, 15, and 25 mmol l-1, and 8 patients underwent hypoglycaemia of 20 min duration after the blood glucose had been kept at 8 mmol l-1 for 1 h. During hyperglycaemia plasminogen activator activity rose from 214 (11-625) (median, range) to 478 (18-772) units (p less than 0.05) at a blood glucose of 5 mmol l-1 and to 511 (89-816) (p less than 0.05) and 535 (33 976) (p less than 0.05) units at a blood glucose of 15 and 25 mmol l-1, respectively. Cross-linked fibrin degradation products (FDP) were 45 and 53 micrograms l-1 at a blood glucose of 5 mmol l-1 and remained unchanged at higher glucose levels. Fibrinopeptide A was 1.3 (0.6-2.8) nmol l-1 at a blood glucose of 5 mmol l-1, and remained unchanged with hyperglycaemia, being 1.3 (0.9-1.3) nmol l-1 after 1h at 25 mmol l-1. During hypoglycaemia, plasminogen activator activity rose from 155 to 745 units (p less than 0.05) while both fibrinopeptide A and cross-linked FDP remained unchanged. The results indicate that acute fluctuations in blood glucose concentration do not lead to thrombin generation. Additionally, increased fibrinolytic activity measured in vitro is not associated with an increase in cross-linked FDP. This suggests that short-term hyper- and hypoglycaemia do not affect the end-products of the coagulation and fibrinolytic pathways. PMID- 2976650 TI - A nation-wide cross-sectional study of glycosylated haemoglobin in Danish children with type 1 diabetes. AB - A nation-wide screening for HbA1c was carried out in Denmark. Twenty-one paediatric departments treating children with Type 1 diabetes participated. During a period of 4 months 884 children were included, 93% of all those followed at these centres, representing approximately 70-80% of all children and adolescents with Type 1 diabetes in Denmark. Among the children 351 were less than or equal to 12 years and 533 were adolescents between 12 and 18 years. Children less than or equal to 12 years had a HbA1c concentration of 8.9 +/- 1.5 (+/- SD)% and an insulin dose of 0.71 +/- 0.3 U kg-1 24-h-1. For adolescents HbA1c was significantly higher 9.7 +/- 2.0% (p less than 0.001) and insulin dose significantly increased 0.85 +/- 0.3 U kg-1 24-h-1 (p less than 0.001). Normal range for HbA1c is 4.0-6.5 (mean 5.3)% of total haemoglobin. Boys and girls less than or equal to 12 years had similar HbA1c, but girls received 13% more insulin (p less than 0.001). In the group of adolescents, girls had a HbA1c 4% higher than boys (9.9 +/- 2.0 vs 9.5 +/- 2.0%, p less than 0.025), received 11% more insulin (p less than 0.001), and had 6% higher body mass index (p less than 0.001). A weak correlation was found between insulin dose and the HbA1c level (r = 0.29, p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976651 TI - Skin problems in workers using visual display terminals. A study of 201 patients. AB - 201 patients with skin problems caused in their own opinion by working with visual display terminals (VDTs) were referred and examined. 18% claimed that their condition improved overnight and 21% that it did so over the weekend. In 25 patients, the skin problems occurred mainly on the cheek turned towards the VDT. Half the patients had rosacea, but their subjective skin symptoms were generally more severe than those in ordinary cases of rosacea, i.e., severe pain, itching and burning. The rest of the patients mainly had common facial dermatoses such as seborrhoeic eczema, acne vulgaris and atopic dermatitis. The patients' skin types, judged by their response to ultraviolet light, did not differ from those of a control population. 18% had non-specific skin problems, e.g., itching and redness. This latter group of patients most closely resembles those cases previously described amongst VDT-workers. An attempt to correlate skin problems with VDTs' electrostatic field strength was unsuccessful because of difficulties in eliminating it. Of 52 electrostatic shields placed in front of patients' VDTs, 38 were defective and 9 were wrongly earthed. The prevalence of migraine-like headache was 40%, which is much higher than in a control population. There was no great use of cosmetics by the study population. When followed up after an average of 8 months, 2/3 of the patients had fewer skin complaints. The question of whether the prevalence of skin problems in general is higher amongst individuals using VDTs than in a control population is addressed in a current study. PMID- 2976652 TI - Rubber contact urticaria. Allergenic properties of 19 brands of latex gloves. AB - To compare the immediate skin test reactivity of various latex (natural rubber) surgical and cleaning gloves, prick tests were performed on 40 latex-allergic persons, 26 of whom were sensitized by surgical and 14 by cleaning latex gloves. 6/17 surgical gloves tested and 1/2 cleaning gloves caused positive reactions in almost all (over 87%) of the allergic subjects. In contrast, the frequencies of positive reactions to 4 other surgical latex gloves were as low as 8-21%, suggesting that not all surgical gloves are equally allergenic. Control prick tests with 1 synthetic rubber and 1 polyvinyl chloride (PVC) glove were negative in all subjects. 2 surgical latex gloves causing either a high or low number of positive prick tests in allergic subjects were analyzed with high-pressure liquid chromatography (HPLC). Similar allergenic protein fractions were detected, which showed protein peaks at MW 2000, 5000 and 30,000 daltons. Corresponding proteins were detected in 1 latex cleaning glove analyzed and in natural rubber. This result confirms that allergenic proteins persist in various surgical and cleaning latex gloves after manufacture from natural rubber and may cause contact urticaria symptoms in sensitized people. PMID- 2976653 TI - Occupational contact allergy to triglycidyl isocyanurate (TGIC, Tepic). PMID- 2976654 TI - Contact allergy to propyl gallate. PMID- 2976655 TI - Contact allergy to 2,4-dichlorophenylethyl imidazole derivativess. PMID- 2976656 TI - Occupational eczema with photosensitivity due to Japanese sargassum. PMID- 2976657 TI - Localization of atrial natriuretic peptide degrading activity in the rat kidney. PMID- 2976658 TI - Renal peritubular versus luminal uptake of biologically active small proteins. PMID- 2976659 TI - Painful and silent myocardial ischaemia during exercise testing. AB - The study was designed to clarify the clinical and angiographic attributes of patients with exercise induced silent myocardial ischaemia. The sample consisted of 102 patients who had ischaemic exercise electrograms (greater than or equal to 1 mm ST segment depression) and had a significant angiographic stenosis (greater than 50% diameter stenosis), in at least one of the major coronary arteries. In 61 patients (60%) painful ischaemia was observed during exercise (group A) while in 41 patients (40%) only ST depression (silent ischaemia) occurred (group B). The groups were similar with respect to age, sex, history of diabetes and all angiographic parameters including the Gensini coronary score. A history of prior myocardial infarctions was observed more frequently (p less than 0.01) in patients with silent ischaemia (group B). Prior myocardial infarction may be a factor causing silent myocardial ischaemia. PMID- 2976660 TI - Burns of the hands due to ethylene oxide used to sterilize gloves. PMID- 2976661 TI - An unanticipated use of a topical cream. AB - A sixteen-year-old black male, distraught over his acne, injected his face via syringe and needle with a topical cream. Within twelve hours, the injected lesions became red and edematous, and later tender. After treatment with oral prednisone and erythromycin, the lesions resolved. This case serves to illustrate the unexpected misuses of cosmetics, in this instance, a patient anxious only to improve his acne problem. PMID- 2976662 TI - A preliminary study on T lymphocyte subsets in systemic lupus erythematosus. PMID- 2976664 TI - [Damages to normal rabbit retina by hematoporphyrin derivative-photodynamic therapy]. PMID- 2976663 TI - [Cause and treatment of back pain in athletes]. PMID- 2976665 TI - Vancomycin-associated exfoliative dermatitis during continuous ambulatory peritoneal dialysis. AB - Vancomycin is commonly prescribed to patients undergoing continuous ambulatory peritoneal dialysis (CAPD) for catheter-related infections and acute episodes of peritonitis. Although adverse dermatological reactions have been reported secondary to the rapid intravenous infusion of vancomycin, the intraperitoneal route of administration has been used routinely during CAPD without these effects. This case report describes a CAPD patient with systemic lupus erythematosus who developed erythema multiforme that progressed to exfoliative dermatitis during intermittent intraperitoneal vancomycin therapy for a catheter related exit-site/tunnel infection. PMID- 2976666 TI - Occupational contact psoriasis. AB - Occupational psoriasis accounts for about 1.2% of all cutaneous pathologic conditions due to work. It is especially important for subjects with slight signs of psoriasis, since patients already suffering from clear illness spontaneously avoid traumatic activities. Our interest was directed to cutaneous symptoms, signs for identifying subjects predisposed to traumatic dermatitis, also useful in vocational guidance. The necessity of official acknowledgement of such work related dermatological pathologies is suggested. PMID- 2976667 TI - [Benign symmetrical lipomatosis--an occupational disease following exposure to tar?]. AB - A recent publication poses the question of a relationship between workers exposed to tar and their tendency to develop benign symmetric lipomatosis (bsL). As this publication has already caused new problems for the assessment of tar exposure, the side effects of tar on the skin and the pathogenesis of bsL using the available literature are discussed. To date, it is not possible to present a coherent pathogenetic model of tar-induced bsL. PMID- 2976668 TI - [Morphobiochemical studies of adrenaline secretion from the adrenal medulla of guinea pigs in experimental anaphylaxis]. PMID- 2976669 TI - Subtyping of phosphoglucomutase 1 (PGM1) in microstains of different biological materials with subsequent species differentiation without further stain material consumption. AB - Micromethods for subtyping of phosphoglucomutase 1 (PGM 1) in small amounts of biological stain material are described, using an applicator for highly diluted stain extracts. With the aid of this applicator strip blood and semen micro stains as well as single hair-roots could be extracted by electroelution directly on the PGM1 isoelectric focusing gel. Species differentiation was also possible either by radial immunodiffusion using the extract remaining in the applicator strip after isoelectric focusing or by interpretation of the PGM pattern itself. PMID- 2976670 TI - The relationship between pharmacokinetics and pharmacodynamics of enoximone in healthy man. AB - The relationship between the pharmacokinetics and pharmacodynamics of enoximone, a new positive inotropic agent, was investigated in 6 healthy men. The volunteers received single oral and i.v. doses of 3 and 1 mg/kg, respectively, and placebo in a double-blind cross-over trial. Plasma concentrations of enoximone and its sulphoxide metabolite, effects on the corrected electromechanical systole (QS2c), the impedance cardiogram (dZ/dt)/RZ index, blood pressure and heart rate were determined over an 8-h period. Peak effects on QS2c and the (dZ/dt)/RZ index were obtained after approximately 1 h. During the first hour, the cardiac effects lagged behind the high plasma concentrations. Thereafter, the effects on QS2c were closely correlated with the plasma concentrations both of enoximone and its sulphoxide derivative (r greater than or equal to 0.90). The concentration-effect curves of both substances were parallel and were independent of the route of administration. The inotropic activity was not related to the drug level in hypothetical peripheral compartments. The results suggest that determination of plasma enoximone 1 h after administration and thereafter may be useful in assessing the haemodynamic activity of the drug. Should this observation also be present in a clinical situation, plasma enoximone measurement might be a valuable tool in management of patients suffering from heart failure. PMID- 2976672 TI - Three putative 5-HT1A agonists act as 5-HT antagonists in frog sensory neurons. AB - The putative 5-HT1A agonists 8-hydroxy-dipropylaminotetralin (DPAT), buspirone and p-aminophenylethyl-m-trifluoromethylphenyl piperazine (PAPP) were tested for their ability to narrow the action potentials of bullfrog sensory neurons, a serotonergic response observed to be blocked by the 5-HT1A antagonists spiperone and spiroxitrine. Given alone, DPAT, buspirone, and PAPP were ineffective 5-HT agonists. When applied concomitantly with 5-HT, however, they significantly antagonized the original response. Thus, in some systems these putative 5-HT1A agonists function as antagonists. PMID- 2976671 TI - The behavioural effects of 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) in mice. AB - The effects of the 5-HT1A receptor agonist, 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) on the behaviour of mice were studied. 8-OH-DPAT given i.v. in doses greater than 1 mg/kg induced the distinct 5-HT syndrome, including head weaving, hindlimb abduction, forepaw treading and tremor. The 8-OH DPAT-induced behaviour was not affected by the 5-HT depleter, p chlorophenylalanine. Reserpine, which depletes monoamines, significantly decreased the head weaving elicited by 8-OH-DPAT, although it did not reduce the other components of the behavioural syndrome. The non-specific 5-HT receptor antagonist, metergoline, attenuated the 8-OH-DPAT-induced behaviour, while the 5 HT2 receptor antagonist, ketanserin, was without effect. In addition, the 5-HT1A receptor antagonist, spiperone, inhibited the 5-HT syndrome elicited by 8-OH DPAT, while the dopamine receptor antagonist, haloperidol, affected only the head weaving. These results suggest that 8-OH-DPAT-induced behaviour in mice is mediated by the postsynaptic 5-HT1A receptor. PMID- 2976673 TI - NAN-190: an arylpiperazine analog that antagonizes the stimulus effects of the 5 HT1A agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). PMID- 2976674 TI - [Confidence in evidence: 10 years after the death of Maccacaro]. PMID- 2976675 TI - [Tumor mortality in a cohort of glass industry workers]. PMID- 2976676 TI - [Participation in preventive child health services in Santiago, Cuba]. PMID- 2976677 TI - [Thalassemia registry of Sicily]. PMID- 2976678 TI - [Contribution of cost accounting to the efficiency of health services: a study on the analytic laboratories of a Roman hospital]. PMID- 2976679 TI - [Determination of HTLV-III-LAV antibodies in 2 different groups of heroin addicts in Verona in 1981 and 1985]. PMID- 2976680 TI - [The Bormida valley]. PMID- 2976681 TI - [Analysis of epidemiological data using a personal computer: problems and integration of diverse software]. PMID- 2976682 TI - [Electroencephalographic manifestations of the convulsive reaction of rats to the intraventricular administration of quinolinic acid and kynurenine]. AB - The EEG responses to intraventricular administration of quinolinic acid (1.5 and 10 mg), pentylenetetrazole (500 micrograms) and kynurenine (200 and 400 micrograms) were studied. Quinolinic acid was found to produce the convulsive discharges in the EEG 30-60 sec after administration. Similar results were obtained following administration of pentylenetetrazole, the first EEG disturbances being recorded in both cases in the leads from the caudate nucleus and hippocampus. Kynurenine administered intraventricularly failed to change the initial EEG in rats. The possible triggering role of the caudate nucleus in the mechanism of quinolinic acid-induced convulsions is suggested. PMID- 2976683 TI - [The existence on the serotonin carrier of 2 independent sites for the specific binding of trazodone and imizin]. AB - A combined effect of zimelidine or trazodone on (3H)-serotonin uptake by the rat brain synaptosomes in the presence of 120 nM imipramine was studied. Imipramine and zimelidine were found to inhibit (3H)-serotonin uptake by a competitive manner, whereas trazodone was a noncompetitive inhibitor. Zimelidine, but not trazodone, was shown to bind to serotonin carrier at the same site as imipramine. PMID- 2976684 TI - [Mechanism of the vascular action of verapamil and a crown ether derivative]. AB - The effects of verapamil and benzyl-aza-15-crown-5 on pressor reactions of blood pressure and perfusion pressure in the femoral artery after administration of noradrenaline, tyramine, angiotensin amide and stimulation of the femoral nerve were studied in acute experiments on anesthetized cats (pentobarbital sodium, 50 mg/kg). Verapamil (0.5 mg/kg) and benzyl-aza-15-crown-5 (9 mg/kg) suppressed pressor reactions to the nerve stimulation and produced no changes at administration of noradrenaline, tyramine and angiotensin amide. One can suggest that the mechanism of the vasodilating action of verapamil and benzyl-aza-15 crown-5 is due to their inhibitory effect on calcium-dependent release of noradrenaline from terminals of the sympathetic nerves. The search of vasodilating agents in the series of crown-ether derivatives is promising. PMID- 2976685 TI - Dynamic angioplasty--a milling catheter for transcutaneous and intraoperative treatment of vascular occlusive disease. AB - Balloon angioplasty and bypass graft surgery are common procedures for treating vascular occlusive disease. The purpose of this preliminary investigation was to evaluate the effectiveness and the safety of a new catheter system, first introduced by KR Kensey. The system involves a flexible catheter with a high speed rotating tip, driven by an electrical motor. The tip is cooled by a continuous flow of sterile saline containing dextran 40, heparin and urokinase. Radiopaque contrast medium may be infused through the catheter to allow the device to be guided and to detect lesions and to evaluate the efficacy of treatment. The system was applied in three patients with occlusive vascular disease (Stage IIb) and segmental or total occlusion of the superficial femoral artery (SFA). The milling catheter was introduced percutaneously into the common femoral artery and guided to the area of occlusion under DSA control. In two patients total recanalisation was achieved after passage of the milling catheter. In one patient the totally occluded SFA could not be cannulated and a femoro popliteal bypass was performed one week later. Complications such as perforation of the vessel or peripheral embolisation were not observed. Pedal pulses were improved significantly in one patient. Further investigations will be necessary to demonstrate whether the milling catheter can be safely used to revascularise patients with limb threatening peripheral vascular disease. PMID- 2976686 TI - [Geriatric dentistry: the dentistry of the future]. PMID- 2976687 TI - Autosomal dominant keratoderma, ichthyosiform dermatosis and elevated serum beta glucuronidase. AB - Five patients in a large pedigree with autosomal dominant palmoplantar keratoderma and/or an ichthyosiform dermatosis were studied. Biopsies of the keratoderma, a remote hyperkeratotic plaque on an arm and the ichthyosis all revealed the same histopathologic changes including hyperkeratosis with round retained nuclei. All 5 patients had elevated serum beta-glucuronidase activity. The significance of beta-glucuronidase in this and other skin diseases is discussed. PMID- 2976688 TI - Fixed drug eruption with lormetazepam (Noctamid) PMID- 2976690 TI - Controlled trials of physical therapy. PMID- 2976689 TI - Deficits in the production of grip forces in Down syndrome. AB - Down syndrome (DS) and control individuals were examined as to their ability to adapt grip forces to changes in the properties of lifted objects. The DS group generated substantially greater grip forces than the controls in all tests and failed to adapt normally to changes in the frictional properties of the objects. Their greater grip forces were not due to greater slipperiness of the skin of their fingers. These results are consistent with other findings of subtle deficits in DS individuals' use of somatosensory information for controlling movement and posture. PMID- 2976691 TI - Family medicine and the developmentally disabled: a relationship worth building. PMID- 2976692 TI - [Hemodynamic and renal effects of synthetic atrial natriuretic factor (alpha-h ANF) in patients with congestive heart failure]. AB - We studied the effects of alpha-human atrial natriuretic factor (alpha-h-ANF) on diuresis, glomerular filtration rate (GFR), natriuresis, atrial blood pressure (BP), heart rate (HR) and plasmatic levels of aldosterone, renin activity (PRA), antidiuretic hormone (ADH) in 21 patients with congestive heart failure (CHF), (NYHA, class IV) and 15 healthy volunteers. Three different doses (25, 50 and 100 micrograms) of alpha-h-ANF were administered; each subject received only one dose. Protocol was as follows: 2 periods of 30 minutes each before alpha-h-ANF administration; 2 periods of 15 minutes and 3 of 30 minutes each after it. Aldosterone, PRA, ADH and plasmatic level of ANF were measured by radioimmunoassay. In patients with CHF 25 and 50 micrograms of alpha-h-ANF produced a significant increase in diuresis, GFR, and natriuresis but no modification of BP. On the contrary BP decreased significantly after 100 micrograms of alpha-h-ANF, without changes in renal function. In healthy volunteers the effects of alpha-h-ANF appeared simultaneously and to same degree after each dose. Both in patients and healthy subjects alpha-h-ANF administration had little effect on aldosterone, renin and ADH secretion. The effects of alpha-h ANF in patients with CHF may be caused by a lowered receptor sensitivity due to a high level of ANF and to their different haemodynamic status. PMID- 2976693 TI - [Coronary angioplasty in unstable angina. Immediate and short-term results]. AB - We report the results of percutaneous transluminal coronary angioplasty (PTCA) in 67 consecutive patients with unstable angina. Twenty patients had new onset (less than 2 months) angina, 33 patients had crescendo angina and 14 had early postinfarction angina. Fifty-one patients had one-vessel disease, 12 patients had two-vessel disease and two patients had three-vessel disease; two patients had a stenosis of a venous graft. In cases with multivessel disease, we performed only the dilatation of the ischaemia-related vessel identified by morphologic features of coronary lesion and electrocardiographic changes during chest pain. The procedure was successful in 54 cases (80.6%). Seven patients (10.4%) had major complications. Emergency coronary artery bypass graft surgery was performed in 6 cases (8.9%) because of occlusion of the left anterior descending artery; despite emergency operation one patient died and two patients sustained a myocardial infarction. One patient had occlusion of the right coronary artery and inferior myocardial infarction. In all patients in whom angioplasty was successful unstable angina disappeared. At 6 months follow-up there were no infarctions or deaths but 14 of 42 patients (33%) had recurrent angina. Restenosis occurred in 16 of 33 patients (48%) who had repeat coronary angiography. Four patients with recurrence of unstable angina had repeat angioplasty; it was successful in 3 cases. One patient died of refractory cardiac arrest. The mortality rate of 71 procedures performed in 67 patients was 2.8% (2/71) and the overall myocardial infarction rate was 4.2% (3/71).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976694 TI - Experience with a 1.2-mm pneumoperitoneum needle for laparoscopy. PMID- 2976695 TI - [Distribution of the incidence of various pelviscopic (laparoscopic) surgical procedures and their complication rates. A statistical analysis in West Germany 1983 up to and including 1985]. AB - The 3rd German pelviscopy (laparoscopy) statistics includes in all 207,823 laparoscopies from 310 clinics and 41,644 laparoscopies from 121 private practises. This is a percentage of 39% from clinics and 59.3% from private practises. There appeared 429 serious complications in the clinics and in the practises 63, which is a complication rate of 1.97%. In 415 cases (1.66%), a laparoscopy was required. The most important of the complications was injury of the intestinal organs, when putting in the Veress-needle or the optical trocar, followed by injury of the main blood vessels, as well as hemorrhages of the salpinges on detached omental fragments. Where tubal sterilization is concerned, 87.7% of the clinics ans private practises used the bipolar HF technique; 32.4% used the endocoagulation according to Semm. In 8.3% of the cases, the monopolar high frequency method was applied. After sterilization in the clinics, 2.8% intra and extrauterine pregnancies occurred, while in the private hospitals this percentage was 3.3%. The intra- or extrauterine pregnancies occurred mainly after sterilization by the bipolar HF technique or after clip-sterilization. 46.1% of the investigated clinics and 36.4% of private practises, intend to increase the indication of endoscopic operational methods. PMID- 2976696 TI - [Laparoscopic findings in suspected adnexitis]. AB - Between 1976 and 1985 laparoscopy was performed on a total of 768 patients in whom adnexitis was suspected. In accordance with conventional usage they were initially designated as "acute" and "chronic" cases of adnexitis on the basis of clinical criteria. The tentative diagnosis of an active infection was confirmed by laparoscopy in 62.7% of the patients with the clinically acute form and 14.5% of those with "chronic" adnexitis. Although high temperature, leukocytosis, and an increased ESR were more common among patients with laparoscopically confirmed adnexitis, high levels of inflammatory reaction were also found in patients with other conditions. In 164 patients (21%) no pathological findings were found in the genital and abdominal regions. The results confirm the importance of laparoscopy in the diagnosis of adnexitis. PMID- 2976697 TI - [Chlamydia trachomatis--detection of inflammatory diseases of the fallopian tubes]. AB - The prevalence of Chlamydia trachomatis in tubal swabs obtained by laparoscopy was investigated in a prospective study. Specimens were collected from women with salpingitis (group 1), tubal infertility (group 2) and from controls, which were considered as being not infected and subjected to laparoscopy for other reasons (group 3). In the period 12/1980-8/1984 chlamydial infection was diagnosed, by means of the McCoy tissue culture method, in group 1 in 19% (16/85), in group 2 in 4% (10/250) and in group 3 in 0% (0/122). In a second period until 6/1985, positive chlamydial cultures were seen in 2% (1/44), 2% (2/110) and 0% (0/75) respectively, however, a simultaneously used IFT (Micro Trak, Syva Merck) was negative in all cases. In the third period until 12/1986, positive chlamydial cultures were obtained in 12% (9/77), 1% (1/137) and 4% (4/90) respectively. The sensitivity of a simultaneously performed EIA (Chlamydiazyme, Abbott) in women with salpingitis was 67%, the specificity was 97%. The EIA seemed to provide an alternative diagnostic method to culture when evaluating tubal specimens. In an additional trial, chlamydial complement fixation test was positive in 26% of a total of 90 women with tubal infertility (and negative tubal culture) vs. 6% in a control group of pregnant females (p less than 0.001). In conclusion, the reported results support the etiologic role of Chlamydia trachomatis as an important agent in salpingitis as well as in tubal infertility. PMID- 2976698 TI - [Indications and results of diagnostic laparoscopy in the assessment of lower abdominal pain]. AB - Between 1976 and 1986 laparoscopy was performed in 3,584 patients to establish the cause of acute and chronic lower abdominal pain. The most frequent indications for laparoscopy were lower abdominal pain of unknown etiology, adnexitis, and adnexal tumors. The overall level of agreement between the clinical and laparoscopic diagnoses was 56.8%. In 50% of the cases with unknown etiology there was no correlation with the complaints. It also proved impossible to establish any organic cause in 8% to 32% of the other cases clinically diagnosed. The principal advantage of laparoscopy is, that the diagnosis is definite, rendering medication and laparotomy unnecessary in many cases. This applies in particular, with regard to appropriate treatment for lower abdominal pain of unknown etiology with no organic findings, and the treatment of benign ovarian cysts. PMID- 2976699 TI - [A. A. Bogdanov (on the 115th anniversary of his birth)]. PMID- 2976700 TI - [Rpo pathway as a possible recombination mechanism of the interaction between DNA of transducing phages and the Escherichia coli genome]. AB - The conditions affecting recombination of DNAs of transducing lambdoid rifd phages with the chromosome of Escherichia coli K-12 recA in the region of homology were studied. In support to the previously obtained data, the Int system was shown to take no part in the process. The homologous character of recombination interactions and their dependence on efficiency of transcription were demonstrated. It is therefore suggested that recombination takes the Rpo pathway. Similar peculiarities were revealed in the processes of interaction between DNAs of rifd phages as well as of lambdoid phages carrying trp genes and the host genome. A hypothesis is put forward that the Rpo pathway operates, depending on the density of DNA supercoiling. PMID- 2976701 TI - [Medico-genetic study of the Kostroma Region population. VIII. Genetic structure of large subdivided populations and its relation with the incidence of autosomal recessive pathology]. AB - FST was estimated for 67 communities and 6 small towns of Kostroma province. The mean FST value for rural and urban populations was (0.83 +/- 0.08).10(-3) and (0.29 +/- 0.07).10(-3), respectively. The connection between FST values and the load of autosomal-recessive disorders was established; the coefficient of correlation (by Chuprov) was 0.34 (chi 2 = 8.45; P less than 0.05). The matrices of genetic distances for two groups of districts of Kostroma province, based on surnames frequencies, were calculated. Dendrogramms were constructed using genetic distances, which reflect the degree of genetical similarity of the populations. The conclusion drawn from the analysis of these dendrogramms is that there is distinct isolation by distance in populations of Kostroma province. It is shown that genetical subdivision of populations is dependent on geographical and some other factors and on the load of autosomal-recessive pathology in the population. PMID- 2976702 TI - [Medico-genetic study of the Kostroma Region population. IX. Interpretation of the genetic distance matrix]. AB - Two kinds of matrices of genetic distances were compared. The first originate from the distribution of surnames frequencies in communities of some districts of Kostroma province. The others were constructed, based on the population size and the physical distances between populations. Essential similarity in results obtained by these two methods was noted. The differences arise, due to the elements of infrastructure in the real populations which are not taken into account using the second method. PMID- 2976703 TI - [Complex hygienic standardization of the herbicide fenagon in reservoir water and food products]. PMID- 2976704 TI - [Sanitary-hygienic indicators of water supply and the incidence of intestinal infection among the population]. PMID- 2976706 TI - [Osteoarticular changes in vibration disease]. PMID- 2976705 TI - [Effect of polymer materials on the reproductive function of men]. PMID- 2976707 TI - [Hygienic standardization of isobutyronitrile in reservoir water]. PMID- 2976708 TI - [Hygienic characteristics of multi-story buildings under construction in Tashkent]. PMID- 2976709 TI - [Survival of pathogenic microorganisms in snow]. PMID- 2976710 TI - [Experience of the preventive services department in conducting mass screening of workers at a railroad junction]. PMID- 2976711 TI - [Use of gas chromatography in determining the degree of environmental pollution]. PMID- 2976712 TI - [Substantiation of single maximum permissible levels of acrylic and methacrylic acids in the air of populated regions]. PMID- 2976713 TI - [Hygienic substantiation of the maximum permissible level of ambush in reservoir water]. PMID- 2976714 TI - [Characteristics of local overexertion of the hands]. PMID- 2976715 TI - [Functional activity of nonspecific suppressors and indicators of lymphocyte blast transformation in patients with occupational allergy]. PMID- 2976716 TI - [A case of bullous contact dermatitis caused by butifos]. PMID- 2976717 TI - [Hygienic characteristics of the working conditions of operators of chemical shops at coke plants]. PMID- 2976718 TI - [Fixation of the vagina to the rectus abdominis muscle in the treatment of stress induced urinary incontinence in women]. PMID- 2976719 TI - [Pregnancy carried to term following administration of cyproterone acetate in the 1st trimester]. PMID- 2976720 TI - [Prenatal diagnosis of complex vitium cordis in routine ultrasound screening]. PMID- 2976721 TI - [Premature luteinization and its treatment with an LH/RH analog]. PMID- 2976722 TI - [The protective effect of medroxyprogesterone acetate in patients with ovarian cancer treated with chemotherapy]. PMID- 2976723 TI - [Laparoscopic therapy of cystic ovarian tumors]. PMID- 2976724 TI - [Atrial natriuretic factor (ANF)--physiological aspects]. PMID- 2976725 TI - The dynamics of eosinophilia in concurrent infections with Trichinella spiralis Owen, 1835 and Toxascaris leonina Linstow, 1909. AB - The concurrent infection with larvae of Trichinella spiralis and eggs of Toxascaris leonina was studied under various conditions using 75 male white mice. The changes in content of eosinophilic leucocytes in the blood, as well as the total number and distribution of larvae of both parasites in different body tissues were demonstrated. The primary infection with Toxascaris leonina caused an increase in the number of eosinophilic leucocytes from day 4 p.i., whereas the infection with Trichinella spiralis larvae induced an increase only from day 7 p.i. An antagonism was observed between the two parasite species: the primary infection with T. leonina led to a decrease in the total number of muscle larvae of T. spiralis, and, vice versa, the primary infection with T. spiralis suppressed the development of T. leonina. PMID- 2976726 TI - Effect of voice quality on adult's perceptions of Down's syndrome children. PMID- 2976729 TI - The intelligibility of synthesized speech in data inquiry systems. PMID- 2976727 TI - Hemodynamic, renal, and endocrine effects of 4-h infusions of human atrial natriuretic peptide in normal volunteers. AB - A synthetic human atrial natriuretic peptide of 26 aminoacids [human (3-28)ANP or hANP] was infused into normal male volunteers. Six subjects were infused for 4 h at 1-wk intervals with either hANP at the rate of 0.5 or 1.0 microgram/min or its vehicle in a single-blind randomized order. Human (3-28)ANP at the dose of 0.5 microgram/min raised immunoreactive plasma ANP levels from 104 +/- 17 to 221 +/- 24 pg/ml (mean +/- SEM), but it induced no significant change in blood pressure, heart rate, effective renal plasma flow, glomerular filtration rate, or renal electrolyte excretion. At the rate of 1.0 microgram/min, human (3-28)ANP increased immunoreactive plasma ANP levels from 89 +/- 12 to 454 +/- 30 pg/ml. It reduced effective renal plasma flow from 523 +/- 40 to 453 +/- 38 ml/min (P less than 0.05 vs. vehicle), but left glomerular filtration rate unchanged. Natriuresis rose from 207 +/- 52 to 501 +/- 69 mumol/min (P less than 0.05 vs. vehicle) and urinary magnesium excretion from 3.6 +/- 0.5 to 5.6 +/- 0.5 mumol/min (P less than 0.01 vs. vehicle). The excretion rate of the other electrolytes, blood pressure, and heart rate were not significantly modified. At both doses, human (3-28)ANP tended to suppress the activity of the renin angiotensin-aldosterone system. In 3 additional volunteers, the skin blood flow response to human (3-28)ANP, infused for 4 h at the rate of 1.0 microgram/min, was studied by means of a laser-doppler flowmeter. The skin blood flow rose during the first 2 h of peptide administration, then fell progressively to values below baseline. After the infusion was discontinued, it remained depressed for more than 2 h. Thus, in normal volunteers, human (3-28)ANP at the dose of 1.0 microgram/min produced results similar to those obtained previously with rat (3 28)ANP. It enhanced natriuresis without changing the glomerular filtration rate while effective renal plasma flow fell. It also induced a transient vasodilation of the skin vascular bed. PMID- 2976728 TI - Storage of spheroplasts at -70 degrees C for transfection with phi X174 RF DNA. AB - A method was developed for preparation of spheroplasts used for transfection with phi X174 RF DNA. This method had a high-level competence and retained the competence for up to one year of storage in 7% DMSO at -70 degrees C. PMID- 2976730 TI - A PGM1*1A variant with a reduced activity. AB - An adverse homozygosity at the phosphoglucomutase-1 (PGM1) locus was detected in a family by electrophoresis on starch gels. Isoelectric focusing on polyacrylamide gels could clearly demonstrate a faint band at the same position as that of PGM1 1A in the mother and the child. Further analysis of other family samples and densitometric evaluation of the stained bands revealed the genetic transmission of a variant allele with a reduced activity. The molecular basis for the low activity is still unknown. PMID- 2976731 TI - In vitro functional characterization of SAG-3--a naturally occurring monoclonal antibody in NZB mice: specificity toward functional T cell subsets. AB - Natural thymocytotoxic autoantibodies (NTA) have been suggested to contribute to the T cell mediated immune dysfunction prevalent in NZB mice. However, direct studies to confirm these suggestions have been difficult to perform due to the absence of large amounts of pure NTA. To overcome this problem, we generated a naturally occurring monoclonal antibody from an NZB mouse which possesses the serological characteristics of serum-NTA. Herein, we report the in vitro functional characteristics of this monoclonal antibody designated SAG-3. In vitro treatment of spleen and lymph node cells from BALB/c mice with SAG-3 and complement eliminated most of the proliferative response to Con-A and moderately reduced the response of these cells to PHA-P and PWM. Complement mediated cytolysis of SAG-3 reactive cells significantly reduced the capacity of unprimed lymph node cells to generate primary CTL responses in vitro against allogeneic cells. However, only a partial decrease in CTL activity was observed when primed secondary CTL effector cells were treated, due primarily to the depletion of CTL's specific towards MHC class I antigens. Furthermore, a moderate reduction in antigen-specific helper T cell function was observed. Pretreatment of responder cells with SAG-3 + C' did not appear to reduce the proliferative MLR response to a wide range of stimulator cells. In addition, contrary to previous reports on serum-NTA, SAG-3 appears not to be reactive against Con-A induced suppressor T cells. Finally, by FACS analysis SAG-3+ thymocytes constitute an overlapping population of Lyt-2+ and L3T4+ thymocytes. These findings are discussed in comparison to previous reports on NTA. PMID- 2976732 TI - Regulation of active transport of monovalent cation across the animal cell plasma membranes by cytosolic regulatory proteins. PMID- 2976733 TI - Effect of hypertonicity and monensin on CD3/TCR surface expression in human T cells. AB - Pretreatment of T lymphocytes with anti-CD3 or anti-TCR mAb results in the disappearance of the T cell receptor complex (TCR/CD3) from the cell surface. The mechanisms of this down-regulation have not been fully elucidated. We demonstrate here that the modulation of the CD3/TCR complex can be completely inhibited by hypertonic medium, a condition known to block receptor-mediated endocytosis. Consequently, the sequestration of the CD3/TCR complex associated to relevant mAb in acid vesicles did not occur under hypertonicity condition. Moreover, monensin, which is known to interfere with receptor recycling, was found to amplify the CD3/TCR modulation induced by specific mAb and decrease the uptake of 125I labeled anti-CD3/TCR mAb by T cells. We therefore propose that mAb-CD3/TCR complexes are internalized via coated pits by a receptor-mediated endocytosis and then transported to acid compartments. MAb are degraded in lysosomes during this process, whereas CD3/TCR molecules recycle back to cell surface. PMID- 2976734 TI - Regulation of allospecific suppressive cell generation in vitro. AB - Control of the generation of suppressive cells (SC) in allogeneic mixed lymphocyte cultures (MLC) has been re-investigated. Cells taken from 3-6 days old ("early", e-MLC) suppressed lymphocyte proliferative responses when transferred to a second MLC. Suppression was not allospecific, nor restricted to the autologous responder. In contrast, cells from MLC 7-10 days old ("late", l-MLC) mediated weak but specific suppressive activity, which was HLA-restricted. Addition of early MLC cells (e-MLC1) to a second MLC (MLC2), blocked the further generation of non-specific suppressive activity in that MLC, suggesting the presence of downregulatory components for SC generation. Nonetheless, the generation of allospecific suppression in late MLC2 (l-MLC2) was enhanced rather than inhibited under these conditions. In separate experiments, a T cell clone (TCC) was isolated which mimicked the action of e-MLC cells on SC generation, namely, down-regulation of non-specific but amplification of specific SC generation. This clone carried the "alternative" tau/delta T cell receptor. Thus, tau/delta+ "antisuppressor" regulatory cells may have a role in the establishment and maintenance of allograft tolerance. PMID- 2976735 TI - Binding of immunoglobulin G to peripheral blood lymphocytes. AB - The specific and saturable binding of FITC conjugate of aggregated goat IgG to goat peripheral blood lymphocytes was studied in PBS containing 1% BSA. The polar nature of the specific interaction of heterologous aggregated IgG, IgG monomer and its fragment F(ab'2) with the cells was studied by ELISA using the peroxidase conjugated F(ab'2) of anti-human IgG under different conditions of pH and ionic strength. PMID- 2976736 TI - Nervenkitt: notes on the history of the concept of neuroglia. AB - The evolution of concepts concerning the identity and the functions of neuroglia is traced. Some of the main ideas in the works of Virchow, Deiters, Golgi, Lenhossek, Lugaro, Ramon y Cajal, del Rio-Hortega, Achucarro, Penfield, and others are highlighted. PMID- 2976737 TI - Plasticity of astroglia: evidence supporting process elongation by "stretch". AB - The structural plasticity of cerebral astroglia was investigated in vivo by implantation experiments. Immunocytochemical markers for glia filament protein were used to identify the astrocytes. First it was established that implanted nitrocellulose filters provided a substrate for astrocytes from different brain regions of young rats. Astrocytes attached to the filter and projected fine processes into it. Longer implantation times increased the density and length of glial processes within filter spaces. Astrocytes that penetrated the filters implanted in the tectum exhibited more processes than those in the cortex, suggesting regional differences of astrocyte distributions. Second it was observed that astrocytes that attached to the filter formed elongated processes when they were tethered within an expanding matrix. This was shown by implanting the nitrocellulose filter together with PC12 cells that continued to grow. The implantation of neither PC12 cells without filters nor nitrocellulose filters alone induced the formation of elongated astroglia with parallel aligned processes, resembling radial glia. Such glial forms only occurred in the filter/PC12 cell cografts. This indicates that processes of astrocytes adherent to nitrocellulose filters could be stretched in response to expansion of the surrounding tissue. PMID- 2976738 TI - Plasma membrane of cultured oligodendrocytes: I. Isolation, purification, and initial characterization. AB - Oligodendrocytes generate myelin as extensions of the plasma membrane. Myelin has been well characterized, yet little is known concerning oligodendrocyte plasma membrane. We have developed a reproducible method for the isolation of an oligodendrocyte plasma membrane-rich fraction (F2.2). F2.2 has a 25-fold enrichment in K+-dependent p-nitrophenyl phosphatase, a plasma membrane marker. Impurities are composed of Golgi elements (8-12%), microsomes (4-6%), and lysosomal membranes (1-5%). Our starting material was oligodendrocytes kept in culture in a nonattached state for 3 to 5 d. After disrupting the cells and removing nuclei (P1), the supernatant (SP1) was fractionated on a self-generating 20% Percoll gradient into three bands: F1, F2, and F3. F1 had only 3% of the applied protein and was not characterized. F2, with 11% of the protein, was fivefold enriched in plasma membrane. F3 had 27% of initial protein; it consisted of a crude mitochondrial and lysosomal fraction. F2 was further purified by first washing it hypotonically, treating it with Mg2+, and then fractionating it on a Ficoll step gradient that yielded F2.2 at the interphase. Morphologically F2.2 comprises 1) membranous sheets, often with more than one membrane in close apposition; 2) membrane vesicles of various sizes and shapes frequently filled with amorphous material; 3) Golgi elements; and 4) unrecognizable profiles. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profile of F2.2 reveals CNPase as a major component in agreement with the high CNPase specific activity (3,860 mumol/mgP/h) found in F2.2. Other significant polypeptides have Mr = 170,000, 135,000, 108,000, 80,000, 53,000, 38,500, 32,000, and 22,200. PMID- 2976739 TI - Plasma membrane of cultured oligodendrocytes: II. Possible structural and functional domains. AB - An oligodendrocyte plasma membrane-rich fraction, F2.2, was resolved by equilibrium density centrifugation on a linear sucrose gradient from 0.5 M to 1.3 M into three fractions, F2.2a, F.2.2b, F2.2c, and a pellet F2.2p. F2.2a and F.2.2b were enriched 1.5-fold relative to F2.2 in plasma membrane markers at the expense of F2.2c and F2.2p, which became correspondingly impoverished. This gave F2.2a and F2.2b a 42-fold and 37-fold enrichment, respectively, in plasma membrane markers relative to the initial cell homogenate. F2.2c had a sevenfold enrichment in a Golgi marker; together with F2.2p, they contained all the Golgi marker initially present in F2.2. Preliminary data indicated that the F2.2 subfractions differed from one another in their molar ratios of cholesterol to phospholipids and protein to lipids but had similar protein profiles when examined by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Their content of fucosylated glycoproteins appeared also to be different. Morphologically, F2.2a and F2.2b were very similar: they contained large membrane vesicles, membrane sheets, and vesicles entrapped within other vesicles. Membrane membrane interaction was apparent in these fractions. F2.2c had many of the same elements, but most of the membrane structures contained amorphous material. F2.2p differed morphologically from the other fractions in that it had principally electron-dense structures. It is postulated that F2.2a, F2.2b, and perhaps F2.2c represent different domains of oligodendrocyte plasma membrane. Alternatively, these fractions might correspond to the plasma membrane of oligodendrocyte subtypes. PMID- 2976740 TI - Astrocytes in the guinea pig, horse, and monkey retina: their occurrence coincides with the presence of blood vessels. AB - In the present study the distribution of astrocytes in the nerve fiber layer (NFL) has been studied in the sparsely vascularized retinae of the guinea pig and horse and in the richly vascularized retina of the Old World monkey (Cercopithecus aethiops) using immunocytochemical methods. In the guinea pig retina glial fibrillary acidic protein (GFAP)-positive astrocytes could not be detected. They were found, however, in the myelinated region of the optic nerve. The optic nerve head and a small retinal region immediately adjacent to it contained few vimentin-positive astrocytes. Histological sections confirmed the restriction of astrocytes to a small retinal region and showed that this is also the only retinal area that is vascularized. Astrocytes showing GFAP and vimentin immunoreactivity were absent from most of the horse retina. They were found only in a narrow zone close to the optic disc, which is also the only region of the horse retina that is vascularized. Thus, as in the rabbit retina (Schnitzer: J. Comp. Neurol. 240:128-142, 1985), in the guinea pig and horse retina astrocytes are not present ubiquitously in the NFL but coexist with blood vessels. In the monkey retina, GFAP-positive astrocytes were found ubiquitously in the NFL. Astrocytes were absent from the avascular foveal region only. It is suggested that the concurrence of retinal astrocytes and intraretinal vascularization may be a feature common to many, if not all, mammalian species. PMID- 2976741 TI - Increased glial fibrillary acidic protein synthesis in astrocytes during retrograde reaction of the rat facial nucleus. AB - Glial fibrillary acidic protein (GFAP) increases in astrocytes following axotomy of facial motoneurons. In the present study we quantified GFAP synthesis both in regenerating facial nuclei after nerve crush and in nonregenerating facial nuclei after nerve resection. An increase in GFAP synthesis during regeneration occurs as early as 24 h after the axotomy. Thus, the increase in the astrocytic GFAP synthesis seems to be the earliest glial response to retrograde changes in facial motoneurons. PMID- 2976742 TI - Brain astrocytes express region-specific surface glycoproteins in culture. AB - Astrocytes derived from the mouse brain mesencephalon and striatum regulate neuronal morphogenesis in a region-specific manner in vitro. To begin defining molecular mechanisms that may underlie this functional heterogeneity, lectin probes were used to compare surface glycoproteins expressed by astrocytes from different brain regions. These experiments demonstrated marked differences in surface glycoproteins depending on the anatomic origin of the astrocytes. In particular, mesencephalic and cerebellar astrocytes express a fucosylated glycoprotein with an apparent molecular weight of 190 kD that is absent or rarely expressed by striatal or cortical astrocytes. These findings raise the possibility that carbohydrate diversity of astrocyte surface molecules may play a role in the heterogeneity of region-specific neuron-glial interactions. PMID- 2976743 TI - Hormone levels in male ice hockey players during a 26-hour cup tournament. AB - Pituitary, adrenal and testicular hormones and steroid-binding proteins were analysed in 15 active ice hockey players and in 16 spectators, matched for physical fitness, during a 26-h cup tournament including three matches. Diurnal variations were noted for all compounds except FSH and SHBG. The levels of cortisol, dehydroepiandrosterone (DHA), 4-androstene-3, 17-dione (A-4) and prolactin increased in players but not in spectators following the matches played in the afternoons, but not following the match played in the morning. Albumin and DHA sulphate (DHAS) levels increased in players following all three matches and SHBG levels increased slightly in players following the afternoon matches, all probably due to haemoconcentration. DHAS and albumin showed simultaneous diurnal variations with a nadir in the morning. 17 alpha-hydroxyprogesterone (17-OHP) and total and non-SHBG-bound testosterone showed similar diurnal patterns in spectators, with maximum levels in the morning. Levels of 17-OHP were largely unaffected by physical stress, while the morning maximum for total and non-SHBG bound testosterone was completely abolished in players. Minor or insignificant changes were noted for LH and FSH. While the changes in unconjugated adrenal steroids (cortisol, DHA and A-4) reflect changes in acute ACTH stimulation, changes in albumin binding may be more important for the short-term variations in levels of DHAS. The influence of physical stress upon the pattern of testosterone may be mediated mainly by inhibition of testicular steroidogenesis by elevated cortisol and/or prolactin levels, rather than by decreased gonadotrophic stimulation or by alterations in binding to SHBG. PMID- 2976745 TI - Cosmetics. Content and function. PMID- 2976744 TI - An investigation of the immune suppression induced by some saikosaponin derivatives. AB - The relationship between lymphocyte activation by purified plant glycoside and the suppression of antibodies against sheep erythrocytes was investigated by an adoptive cell transfer system. Glycosides are one of the main components of Shosaikoto--famous hematopoietic remedies of oriental medicine. In earlier reports, BALB/c mice treated with these drugs, suppressed the plaque-forming cell (PFC) response to T-dependent antigens. In contrast, the PFC response was enhanced for T-independent antigens. In this communication, an experiment was designed to determine the nature of the suppressive factor(s) produced in treated mice. Mice were orally or intraperitoneally administered 1 mg/kg of five types of purified SS derivatives every other day on five consecutive occasions. Spleen cells and serum were prepared from SS-treated mice and were then passively transferred to recipient mice. On the spleen cells (and not any of the sera) from drug-treated hosts effectively suppressed the PFC in recipients. The spleen cells of treated mice were further divided into adherent and non-adherent cells; each cell type was transferred into recipient animals. The observed suppressive activity was clearly detected in the non-adherent cell population. A subsequent analysis of the lymphocyte subpopulation revealed that Lyt.1.1 negative and Lyt.2.2 positive cells were responsible for this suppression. It was concluded that some SS derivatives elevate the activities of the major immunocyte population, with suppressor cell activity predominating. PMID- 2976746 TI - Topical estrogens. Current status. PMID- 2976747 TI - Differential attitudes of Chinese students toward people with disabilities: a cross-cultural perspective. AB - The purpose of this study was to examine the attitudes of 338 Chinese secondary school students toward three major disability groups (people who are physically disabled, emotionally disturbed, and mentally retarded) using an adaptation of the American Scale of Attitudes toward Disabled Persons (SADP). We found that physically disabled persons were rated higher across the three subscales of the SADP compared to either emotionally disturbed and mentally retarded individuals, who were rated similarly low by the students. The lack of differentiation between the two mental handicapping conditions is at variance with contemporary western findings. The overall negative attitudes toward people with mental disabilities may have significant implications for community rehabilitation programming for this population. PMID- 2976748 TI - [Urticaria, abdominal pain, arthritis, pseudotumor cerebri and nephrotic syndrome in a 20-year-old patient]. PMID- 2976749 TI - Pachydermoperiostosis and acro-osteolysis. AB - The authors present a case of pachydermoperiostosis (digital hippocratism, periostosis, pachydermia) associated with acro-osteolysis. This is the first association of this type described in Italy, and it is only recently that foreign authors have reported the presence of acro-osteolysis in patients affected with pachydermoperiostosis. The results obtained from studies of the micro and macro circulation in pachydermoperiostosis are reported. PMID- 2976750 TI - Abilities Expo '88: the exhibition par excellence. PMID- 2976751 TI - [Ghirlandajo's painting "Grandfather and Grandchild" from the viewpoint of the dermatologist]. PMID- 2976752 TI - [Transdermal therapeutic systems (TTS)]. AB - The technique, pharmacokinetics, and advantages and disadvantages of transdermal drug delivery systems (TDS) are discussed. Special emphasis is laid on the dermatological problems arising during use of the TDS, such as irritation, sensitization, and the effects of occlusion, dermatoses, skin microflora and epidermal metabolism. PMID- 2976753 TI - [Epicutaneous testing with a standard series. Results in 12,026 patients]. AB - From 1977 to 1983, 12,026 patients were given patch tests with a standard series of substances. A total of 4,494 (37.4%) had positive patch-test reactions to one or more allergens; the average number of positive reactions was 1.85. The most frequent reactions were due to nickel sulphate (positive reactions in 9.2% of those tested), fragrance mix (8.9%), balsam of Peru (6.3%), cobalt chloride (4.7%), potassium dichromate (4.3%) and wool alcohols (4.3%). Women showed significantly more positive reactions than men to nickel sulphate, cobalt chloride, wool alcohols, and parabens and significantly fewer positive reactions than men to potassium dichromate, PPD mix, and epoxy resin. Younger patients showed significant more positive reactions to nickel sulphate and cobalt chloride; middle-aged patients reacted more to potassium dichromate, paraphenylendiamine, formaldehyde and thiuram mix; elderly patients reacted to balsam of Peru, wool alcohols, caine mix, neomycin sulfate, benzocaine, colophony, clioquinol, mafenide, parabens, and gentamycin sulfate. PMID- 2976754 TI - In vivo transfer of pAM beta 1 from Lactobacillus reuteri to Enterococcus faecalis. AB - Trials were conducted to determine the in vivo transferability of plasmid mediated antibiotic resistance between two strains of enteric Gram-positive bacteria. Germ-free mice were associated with the donor Lactobacillus reuteri DSM 20016 strain, carrying the broad host range pAM beta 1 plasmid, and with the Enterococcus faecalis JH2SS recipient strain. Analysis of faecal content of associated mice demonstrated that the in vivo transfer of this plasmid did occur and that frequencies of conjugation were affected by the presence of subtherapeutic levels of antibiotic in the diet. PMID- 2976755 TI - Classification of handicaps in 6-7-year-old mentally retarded children. Usefulness and inter-observer agreement of a child adapted handicap code of WHO's ICIDH. AB - This paper reports on the clinical use of a classification of handicaps in 6-7 year-old mentally retarded children. A child-adapted handicap code of WHO's ICIDH was used. It was easy to manage and added valuable information to the child's medical status and psychosocial evaluation. The inter-observer agreement was most satisfactory. The handicap dimension of 'social integration' ought to be regarded as 'social interaction' and needs to be specified further. PMID- 2976757 TI - Purification and characterization of beta-N-acetylhexosaminidase I from human placenta. AB - beta-N-Acetylhexosaminidase (hexosaminidase) I, which has an intermediate charge character between those of hexosaminidases A(alpha beta 2) and B[beta beta)2), was purified 1,500-fold from human placenta by procedures including chromatographies on concanavalin A (Con A)-Sepharose and an immunoadsorbent column. The isolated hexosaminidase I was heat-stable, and antigenically cross reactive to anti-beta chain-IgG but not to anti-alpha chain-IgG. The results of substrate specificity experiments using 3H-labeled natural substrates indicated that the hexosaminidase I hydrolyzed Gb4Cer to Gb3Cer but not GM2 to GM3. The tryptic peptide map of the hexosaminidase I was similar to that of hexosaminidase B, though some differences were observed. The hexosaminidase I after treatment with neuraminidase or endo-beta-N-acetylglucosaminidase H was partly converted to less acidic forms. Treatment of the hexosaminidase I with acid phosphatase did not change the charge character. Therefore hexosaminidase I is an acidic variant form of hexosaminidase B, possibly resulting from sialylation and the presence of phosphodiester bonds at the carbohydrate moiety. PMID- 2976756 TI - Ca2+ uptake in bovine adrenocortical microsomes: formation of phosphorylated intermediate of Ca2+ dependent ATPase. AB - Bovine adrenocortical microsomes were prepared and partially purified by discontinuous sucrose density gradient. Light fractions of the microsomes at the interface between 15 and 30% sucrose solution, exhibited ATP dependent Ca2+ uptake. The Ca2+ uptake was dependent on temperature and stimulated by free Ca2+ (the concentration for half maximal activation = 1.0 microM) and Mg2+. The Ca2+ uptake was inhibited by ADP but not affected by 10 mM NaN3 or 0.5 mM ouabain. Calcium release from the microsomes was accelerated by a Ca2+ ionophore, A23187, but not by a Ca2+ antagonist, diltiazem. A microsomal protein with a molecular weight of 100-110 kDa was phosphorylated by [gamma-32P]ATP in the presence of Ca2+, and the Ca2+ dependency was over the same range as the Ca2+ uptake (the concentration for half maximal activation = 3.0 microM). The phosphorylated protein (EP) was stable at acidic pH but labile at alkaline pH and sensitive to hydroxylamine. The rate of EP formation at 0 degrees C in the presence of 1 microM ATP and 10 microM Ca2+ (half time = 0.2 s) was less than that in the sarcoplasmic reticulum (SR) of rabbit skeletal muscle (half time = 0.1 s). The rate of EP decomposition at 0 degrees C after adding EGTA was about 6.7 times slower (rate constant: kd = 4.3 X 10(-3) s-1) than that of SR. It was suggested that adrenocortical microsomes contain a Ca2+ dependent ATPase which function as a Ca2+ pump with similar properties to that of SR. PMID- 2976758 TI - Diabetic complications: is there a way out of the labyrinth? PMID- 2976759 TI - Diabetic nephropathy: natural history and declining incidence in diabetic children. AB - This report reviews data regarding diabetic nephropathy and proliferative retinopathy (PR) that were derived from three inception cohorts of insulin dependent diabetics (IDDM) under the age of 21 years at the onset of diabetes mellitus in the index years 1939, 1949, and 1959. Nephropathy occurred in only a subsegment of this population and was age dependent rather than dependent on the duration of diabetes. It occurred earlier and was more aggressive in those with poorer control of the diabetes. The cumulative incidence of nephropathy decreased significantly between 1939 and the 1949 and 1959 cohorts. Possible reasons for this decline are discussed. Similarities and differences between the occurrence of nephropathy and PR are reviewed. The findings suggest that different etiologic factors are involved in the pathogenesis of these two microvascular complications. PMID- 2976760 TI - Rational drug treatment of the hypertensive diabetic with nephropathy. AB - Development of hypertension in Type I diabetics parallels evolution of nephropathy. In Type II diabetics, excessive prevalence of hypertension prior to the appearance of proteinuria suggests that factors other than nephropathy are operative in its pathogenesis. On the other hand, the risk of nephropathy in Type II diabetics is higher than previously appreciated. Recent evidence suggests that angiotensin II plays an important role in the induction and progression of diabetic nephropathy. This provides a rationale for antihypertensive therapy with converting enzyme inhibitors in nephropathic diabetics in whom they have been shown to lower blood pressure and diminish proteinuria. Furthermore, in a retrospective study of patients with various renal diseases (including diabetic nephropathy), the authors found suggestive evidence that converting enzyme inhibitors may also attenuate progression of renal failure to a greater extent than other antihypertensive drugs. PMID- 2976761 TI - Does the severity of diabetic retinopathy predict pregnancy outcome? AB - The authors sought to determine whether the severity of diabetic retinopathy is a predictor of subsequent pregnancy outcome. One hundred and seventy-nine pregnant diabetic women were evaluated in their first trimester of pregnancy. Stereoscopic color photographs of the ocular fundus were taken and graded by the Fundus Photography Reading Center. Thirty-nine women had no retinopathy, while 28 had proliferative retinopathy in the worse eye. The women's history and hospital delivery room charts were reviewed with regard to pregnancy outcome. Thirty-three pregnancies terminated with an adverse outcome. A logistic regression analysis was used to evaluate significant predictors of pregnancy outcome. Of maternal age, duration of diabetes, glycosylated hemoglobin, proteinuria, cigarette smoking status, and severity of diabetic retinopathy, only the last variable significantly predicted an adverse outcome. These data suggest that the severity of retinopathy should be considered when counseling a pregnant diabetic woman. PMID- 2976762 TI - Some lessons from the studies of renal biopsies in patients with insulin dependent diabetes mellitus. AB - The authors have studied relationships of renal structure and function in more than 100 patients with insulin-dependent diabetes mellitus (IDDM), aged 13-55 years (mean, 30 years) with diabetes for 1-30 years (mean, 19 years). The authors confirmed the unique nature of the diabetic lesions that, in constellation, occur in no other disease. It was found that increased fractional mesangial volume (Vv Mes) is strongly associated with decreased glomerular filtration rate (GFR), proteinuria, and hypertension and that all patients with overt diabetic nephropathy have Vv Mes in excess of 0.35 micron 3/micron 3. This relationship results from constriction of the capillary lumen and filtration surface as a consequence of increased Vv Mes. Global glomerulosclerosis (scarring) is common in IDDM patients and appears related to arteriolar hyalinosis. Focal segmental glomerulosclerosis is a rare lesion in these patients. Having a single kidney (transplanted IDDM patients) is not associated with accelerated lesion development. The presence or absence of microalbuminuria (MA), per se, does not predict underlying glomerular structure, which may vary from the normal range to a level of pathology bordering on that regularly associated with overt nephropathy. However, when MA is associated with hypertension, or reduced GFR or both, urine albumin excretion (UAE) generally exceeds 40 mg/24 hr, and glomerular pathology is always present. The authors concluded that diabetic nephropathy is a unique renal disorder that cannot be caused by hemodynamic factors alone. The authors further conclude that MA becomes a predictor only when other features of overt nephropathy are already present and that serious diabetic glomerular lesions can be present in patients with normal UAE. PMID- 2976763 TI - Diabetic cystopathy--a risk factor in diabetic nephropathy? AB - Twenty-seven patients with Type I diabetes and diabetic nephropathy were repeatedly tested (mean, 6 times per patient) for residual urine volumes with a noninvasive technique. Results in 43 of 162 investigations (27%) were abnormal, with residual volumes of greater than 15 ml. Twelve of 162 (7%) showed a residual volume greater than 100 ml. In any individual patient the occurrence of residual volumes was not a consistent finding, and the volumes varied. Pathologic residual volumes were more common in men, but all those with bacteriuria were women. All patients with residual volumes (N = 16) were given voiding instructions. There was no increase in residual volumes during the observation period (mean, 32 months), the median residual volume being 8 ml at the first observation and 5 ml at the last observation. The occurrence of residual urine could not be shown to correlate with progression of renal insufficiency. It is suggested that all patients with long-standing Type I diabetes should be tested by a noninvasive technique for residual urine volume and given voiding instructions to avoid acute retention episodes and complete atony of the bladder. PMID- 2976764 TI - Atrial natriuretic factor in various stages of diabetic nephropathy. AB - Plasma concentrations of atrial natriuretic factor (ANF) were measured by radioreceptor assay in 148 insulin-dependent Type I diabetic patients with different stages of diabetic nephropathy. In patients with overt or incipient diabetic nephropathy as well as in patients with glomerular hyperfiltration only, ANF concentrations were elevated when compared with non-nephropathic diabetic and normal subjects. No correlations between ANF and blood pressure, age, diabetes duration, or parameters of metabolic control were found. These findings may indicate a pathophysiologic role of ANF in the development and progression of diabetic nephropathy. PMID- 2976765 TI - Triglyceride levels affect cognitive function in noninsulin-dependent diabetics. AB - Noninsulin-dependent diabetes mellitus (NIDDM) is associated with decrements in several cognitive functions. Among the variables that apparently contribute to the decline in cognitive performance is poor glucose control, as measured by hemoglobin A1c. Elevated levels of triglycerides in diabetics may also contribute to this cognitive decline through the increased incidence of atherosclerosis in these patients. The authors examined the relationship between triglycerides and cognitive performance in 246 NIDDM outpatients, aged 55-74 years. The relationship between triglyceride levels and performance on three cognitive tasks and on a test of reaction time was measured. Elevated levels of triglycerides were associated with significant decrements in performance on the digit symbol substitution test, digit span (backward) test, and on a reaction time measure. High levels of triglycerides, independent of chronic glucose control, appear to contribute to the decreased ability to perform short-term memory tasks in NIDDM. PMID- 2976766 TI - Nonenzymatic glycation: a central mechanism in diabetic microvasculopathy? AB - Nonenzymatic glycation of proteins is one of several theories advanced in recent years to explain the pathogenesis of complications of diabetes. It is a condensation reaction between glucose and free amino groups, and the extent of glycation is largely dependent on the glucose concentration to which the protein is exposed and the biologic half-life of the involved protein. Because the lesions of diabetic microangiopathy involve basement membranes, many of the relevant studies have examined the consequences of excess glycation in collagen and other extracellular matrix proteins. Additionally, recent work indicates that glycated proteins, through a series of rearrangement reactions, give rise to abnormal cross links and complexes that are believed to alter structure-function properties and to have pathophysiologic importance. From these studies, a two tiered scheme for the natural history of nonenzymatic glycation is proposed. This scheme is clinically relevant, particularly with respect to diabetic management programs. PMID- 2976767 TI - Demographics, diabetes type, and degree of rehabilitation in diabetic patients on maintenance hemodialysis in Brooklyn. AB - A point prevalence study of 232 uremic diabetics undergoing maintenance hemodialysis was conducted at fourteen facilities in Brooklyn, NY, to ascertain extent of rehabilitation, diabetes type, and immediate family history of diabetes according to the diabetes type in the proband. The majority of patients were black (138, 59%) and female (131, 56%). When grouped by diabetes type, insulin dependent (Type I) diabetics were a small minority (31, 13.4%) of the total study population. With the exception of those with onset of diabetes in childhood, there was no difference between the interval between diagnosis of diabetes and development of renal failure in Type I (15.3 +/- 8.6 years) and the overall group of diabetics (14.9 +/- 9.3 years). A history of diabetes in an immediate family member was found in 114 (49.1%) of the entire group and was approximately the same in Type I (41.9%) and Type II (52.5%) diabetics. Rehabilitation was poor for the group as a whole, with a mean Karnofsky score of 64.9 +/- 14.3, which is a level indicative of the need for assistance in everyday living, and there was an inverse correlation between increasing age and declining Karnofsky score. Factors inhibiting rehabilitation included serious vision loss in 137 (59.1%) subjects, limb amputation, and prior myocardial infarction and stroke. Only 7 of the 153 patients (4.8%) of those younger than age 65 were gainfully employed outside of the home, and only 27% of surveyed patients were able to attend to activities beyond self care. Maintenance hemodialysis, although life extending, does not induce substantive rehabilitation for the uremic diabetic. PMID- 2976769 TI - Issues in the classification of mental retardation: differentiating among organic etiologies. AB - Many researchers of mental retardation fail to take etiological differences into account, although the value of distinguishing between organic and familial mental retardation has long been discussed. The argument is made that even the two-group approach needs to be extended so that groups with different organic etiologies are studied separately. Taking etiological distinctions into account will allow for more precise research and a better understanding of mental retardation. Evidence for the utility of differentiating retarded persons by etiology is provided by a research review showing examples of behavioral differences between organically retarded groups. It is concluded that such differentiation also has clear implications for intervention. PMID- 2976768 TI - A survey of the NIH CAPD Registry population with end-stage renal disease attributed to diabetic nephropathy. AB - A survey of CAPD/CCPD patients with end-stage renal disease attributed to diabetes mellitus done by the USA NIH CAPD Registry obtained information from 499 patients. These data suggest that in diabetics with renal insufficiency, the time interval from age at diagnosis of diabetes to initiation of dialysis decreases as the age of diagnosis increases. Mean interval from the time of diabetes diagnosis to CAPD or CCPD initiation was 25 years for patients less than 20 years of age at diagnosis and 17 years for patients greater than or equal to 30 years of age. This trend is independent of the type of diabetes management and appeared to be independent of the type of diabetes. Patients were categorized on the basis of pre- and post-CAPD management of hyperglycemia. There were several associations noted between type of diabetes therapy and clinical findings: A higher proportion of legally blind patients had used insulin only (33%) compared with patients never using insulin (10%) and 78% of patients using insulin only were white compared with 49% among the never on insulin group. This latter result indicates that race influences the type of diabetes and/or progression of diabetes to renal insufficiency. Patients on insulin only reported parents and/or siblings with diabetes less often than did patients using insulin and oral agents, some insulin, or never any insulin to manage their diabetes. The authors also noted that peritonitis rates were not increased in those patients who added insulin to dialysis solutions. PMID- 2976770 TI - Simultaneous microassay for etretinate and its active metabolite, etretin, by reversed-phase high-performance liquid chromatography. PMID- 2976771 TI - Purification of phosphofructokinase using transition-state analogue affinity chromatography. AB - A novel purification of phosphofructokinase has been achieved in a two step process using ion-exchange affinity chromatography and a transition-state analogue affinity column matrix. The procedure can be performed in one day, and gives a 25% yield of the starting material. The transition-state analogue chromatography is carried out using an ADP-agarose column in the presence of fructose 6-phosphate, magnesium ions and nitrate ions. In the presence of nitrate ion plus substrate, phosphofructokinase binds immobilized ADP while other proteins pass through the column. Previous studies with creatine kinase have shown that the nitrate ion mimics the planar phosphate in the transition state resulting in a complex which is stable under the relatively high ionic strength of the column buffer. This permits the elution of phosphofructokinase in a single peak of high specific activity. This column typically results in a 20-30 fold increase in specific activity with only a small loss of activity. PMID- 2976772 TI - Immunoblots, antimicrobial resistance, and bacteriophage typing of oxacillin resistant Staphylococcus aureus. AB - An immunoblotting system was developed for typing of oxacillin-resistant Staphylococcus aureus. Clinical isolates recovered during a 40-month period at a single institution were evaluated with this typing scheme. Results were compared with susceptibility patterns and with bacteriophage typing results for 100 clinical isolates and with plasmid fingerprints for 14 isolates. Immunoblotting was found to be a useful method with good reproducibility that distinguished seven major groups of patient isolates that were clinically and epidemiologically related. Susceptibility patterns showed specific correlations with other typing results but were inferior to immunoblotting and phage typing for differentiating major groups of organisms. Plasmid profiles failed to distinguish two major groups that were readily identified by immunoblots and phage typing. There was evidence of increasing antimicrobial resistance of endemic hospital strains. Immunoblotting correlated well with phage typing, offered an alternative method for typing isolates that could not be typed by phage typing, and was superior to susceptibility testing and plasmid profiles for distinguishing different groups of oxacillin-resistant S. aureus at our institution. PMID- 2976773 TI - Variability of clarithromycin and erythromycin susceptibility tests with Haemophilus influenzae in four different broth media and correlation with the standard disk diffusion test. AB - Four separate laboratories performed antimicrobial susceptibility tests with 40 Haemophilus influenzae isolates, each tested in triplicate. Erythromycin and a new macrolide, clarithromycin (A-56268; TE-031), were tested by the disk diffusion method, by the agar dilution procedure in two different media, and by broth microdilution tests in four different media. Erythromycin MICs for 90% of the strains were 16 micrograms/ml in Mueller-Hinton broth with 3% lysed horse blood and NAD, 4.0 micrograms/ml in hemophilus test medium, and 2.0 micrograms/ml in supplemented Schaedler broth or in the fastidious broth medium from Beckman Instruments, Inc. Clarithromycin MICs were generally 1 doubling dilution greater than erythromycin MICs in each of the media. Erythromycin disk tests corresponded best with MICs determined in the fastidious broth medium. In that same medium, clarithromycin MICs were about 1 doubling dilution greater than what would be expected from the results of disk tests. Because there were fewer growth failures, hemophilus test medium is recommended for microdilution tests with H. influenzae. Incubation of all tests for a full 24 h without an increased CO2 atmosphere was needed to achieve maximal precision of the tests. Interlaboratory and intralaboratory reproducibility of all tests was satisfactory. PMID- 2976774 TI - Comparison of Dacron-tipped applicator and cytobrush for detection of chlamydial infections. AB - Endocervical samples were obtained from 435 females for Chlamydia detection by using both a Dacron swab and a cytobrush. Positive results were obtained from 35 swabs and 34 cytobrush specimens. All specimens positive with the cytobrush were detected also with swab samples. The cytobrush and Dacron swab appear to be comparable for the detection of endocervical infections with Chlamydia trachomatis. PMID- 2976776 TI - Effects of whole cottonseed, niacin, and niacinamide on in vitro rumen fermentation and on lactating Holstein cows. AB - In Experiment 1, effects of whole cottonseed (0, 5, 15, or 30% of the total ration DM) on in vitro ruminal fermentation showed increased ruminal pH and ammonia concentration but lowered microbial protein. Acetic acid concentration was greatest with diets of 15 and 30% whole cottonseed, but propionate and total VFA concentrations were reduced by increasing whole cottonseed from 0 to 30%. In Experiment 2, neither niacin nor niacinamide (0, 100, 200, or 400 ppm) altered substantially fermenter pH or ammonia concentration. Both niacin and niacinamide increased synthesis of microbial protein. Acetate and propionate concentrations were not altered by treatment. Total VFA concentration tended to be lower as concentration of niacin and niacinamide increased. In Experiment 3, 28 Holstein cows were used to determine the effects of supplemental niacin on feed intake, milk yield, and composition. Cows were fed individually complete mixed diets ad libitum containing either: 1) 0; 2) .015; 3) .03; or 4) .06% niacin. There was a trend for lower milk fat test with niacin supplementation. Milk protein percentage was higher without niacin than with niacin at .015 or .03% in the diet, but daily milk and protein yields were higher with .06% versus .015% of niacin. Supplemental niacin did not affect casein nitrogen, lactose or minerals percentage, or concentrations of plasma glucose and insulin. PMID- 2976777 TI - Some environmental factors influencing the breeding of Anopheles balabacensis complex (dirus) in domestic wells in Burma. PMID- 2976775 TI - Benefits of a back care and light duty health promotion program in a hospital setting. AB - Back injuries are a common and expensive problem in industry in terms of both direct and indirect costs. A dramatic increase in the costs of workers' compensation for work-related back injuries occurred from 1981-1985 in a hospital setting. During this five-year period, costs of back injuries rose from $36,384 to $272,751, a 750% increase, and the incidence per thousand employees rose from 21 in 1981 to 33 in 1985. A back care education program was instituted by the Physical Therapy Department and Employee Health Service for all departments within the hospital, with content tailored to individual job requirements. Because employees who have had one back injury are at risk for reinjury, an "at risk" program was also developed. For those injured employees without time off work and without need for medical care, a private session with a physical therapist is required to assess musculoskeletal problems and to review posture and body mechanics directly related to specific job tasks. Those employees unable to work in their usual positions are assigned to temporary modified jobs as part of the Light Duty Program. In 1986, following the institution of these two programs in the hospital, the cost of back injuries dramatically decreased to $72,296, a $200,000 drop, and the incidence per thousand employees fell to fifteen. The efficacy of this health promotion intervention program was confirmed both in the cost benefit to the hospital as well as in reduction in incidence of low back injury. PMID- 2976779 TI - Percutaneous transluminal coronary angioplasty and its role in thrombolysis: an overview of current practice. PMID- 2976778 TI - Ultrastructure and cytochemistry of the tegument of Orthocoelium scoliocoelium and Paramphistomum cervi (Trematoda: Digenea). AB - The tegument of Orthocoelium scoliocoelium and Paramphistomum cervi was examined using histochemical techniques and electron microscopy. On the basis of the distribution of acid and alkaline phosphatase (E.C. 3.1.3.2, E.C. 3.1.3.1), non specific esterase (E.C. 3.1.1.1), cholinesterase (E.C. 3.1.1.7) and succinate dehydrogenase (E.C. 1.3.99.1) at light microscope level two distinct regions were recognized, an outer and an inner zone. Electron microscopy revealed that the tegument comprises an outer surface syncytium underlain by a thick subsyncytial zone and musculature. Deeper still occur the nucleated "tegumental cells". The latter are in cytoplasmic continuity with the surface syncytium via vacuolated cytoplasmic trabeculae which traverse the muscle layers and the subsyncytial zone. Three types of tegumental cells each lacking mitochondria were observed. The T1 cells synthesize discoid and electron dense T1 bodies while T2 cells produce oval and electron lucent T2 bodies. The third type of tegumental cells apparently produce no secretory bodies and may represent an embryonic cell type. The surface syncytium contains T1 and T2 secretory bodies and is bounded apically by a plasma membrane invested externally by a fuzzy and filamentous glycocalyx. The surface syncytium lacks mitochondria and is traversed by infoldings of the basal plasma membrane. Beneath the surface syncytium the subsyncytial zone is largely comprised of fibrous interstitial material. This zone, which is particularly thick in the amphistomes, is traversed by trabeculae and extensions of underlying parenchymal cells which usually contain mitochondria and lysosomes. The subsyncytial zone overlies numerous circular and longitudinal muscle fibres. The absence of mitochondria and enzymes associated with active transport suggests that the amphistome tegument may be mainly specialized for protection of the worm against mechanical and chemical conditions prevailing in the rumen. Active uptake of nutrients is probably not a primary function. PMID- 2976780 TI - Mls and the helper T-cell repertoire. I. Mls as a regulator of T-helper cell activation. AB - We provide evidence that the Mls reaction involves a broad cross-section of the helper cell population. In addition to those cells reacting overtly to Mls stimulatory spleen cells, there is a second large population of helper cells that are affected by an Mls difference. This latent Mls effect is manifested by either synergy or antagonism in mitogen-mediated, Ia-dependent T-cell activation, depending on Mlsa or Mlsb phenotypes of stimulator cells, respectively. Our data can be explained by attributing to Mls alleles the function of differential regulation of autoreactivity of T cells. The results show that the concept of the Mls exerting a negative signal deserves serious consideration. PMID- 2976781 TI - Mls and the helper T-cell repertoire: II. Does the Mls gene product influence the formation of the T-cell repertoire? Studies with Mls tolerance. AB - In the formation of a repertoire, T cells are selected through a window of autoreactivity which is defined by a low boundary representing the minimal autoreactivity required to enter the T-cell compartment and a high boundary which determines the highest admissible autoreactivity. We propose that the Mls gene product down regulates autoreactivity and thus modifies the formation of the T cell repertoire. Given the polymorphism of Mls and the assumption that Mlsb is more inhibitory than Mlsd, it is conceivable that Mlsb (responder) mice admit into their T-cell compartment T cells with higher autoreactivity than Mlsd mice. We suggest that it is this highly autoreactive fraction of Mlsb T cells which responds in the overt Mls response. We show that Mls tolerance eliminates T cells responding in the overt Mls response but not T cells responding in the latent Mls response. This is consistent with the finding that T cells responding in the latent Mls response occur in Mlsb as well as in Mlsd mice. PMID- 2976783 TI - Certification: recognizing excellence. PMID- 2976782 TI - Genetic complexity of Mls. AB - For almost 20 years, little new has been described for Mls or the products it encodes. In the present report, data is presented which indicate that either the number of loci or the number of alleles per locus that control Mls-like products is much larger than the two-locus model or five allele-model presently envisaged. PMID- 2976784 TI - Role delineation and test specification validation study for the CNRN examination. AB - The American Board of Neuroscience Nursing conducted a role delineation and validation study to determine practice areas to be tested. In addition, weighting for each area to be included in the certification examination was determined. This article describes steps taken in the validation process and discusses findings along with application of results to certification examination revision. PMID- 2976785 TI - Wilson's disease: how it changed a young woman's life. AB - Wilson's disease is an inherited disorder which affects the body's ability to metabolize copper. The resulting copper toxicosis can involve all body organs but especially the liver, brain, cornea and kidneys. This article presents a review of the disease and a case study demonstrating how the disease changed a young woman's life, presenting many challenges to her, her family, her nurses and other care providers. PMID- 2976786 TI - An approach to migraine. AB - Since headache is one of the most common symptoms experienced in the general population, the neuroscience nurse should understand the phenomenon of migraine. We define the various types of migraine and present a strategy, based upon precipitators, inhibitors and a "migraine threshold" to facilitate patient education. PMID- 2976788 TI - Progressive supranuclear palsy: nursing care implications. AB - Progressive supranuclear palsy (PSP) is an uncommon, but debilitating, neurological disease. Characterized by paralysis of eye movements, difficulty talking and swallowing, body trunk rigidity, gait ataxia, and dementia, the disease progresses rapidly over a 6-8 year time span. Death usually occurs from aspiration pneumonia. To provide meaningful care for the PSP patient and his/her family, the neuroscience nurse must first understand the disease pathophysiology, then the appropriate care interventions. PMID- 2976787 TI - The Traumatic Coma Data Bank: a nursing perspective, Part II. AB - The Traumatic Coma Data Bank (TCDB) was a collaborative project undertaken to study the nature and course of severe head injury. Evolution of nursing goals includes collection of data, communication, new clinical observations, designing future research projects, education and legislative impact. Management issues inherent in the collaborative research project are defining, collecting, entering, and analyzing and publishing results. PMID- 2976790 TI - Creutzfeldt-Jakob disease. AB - While the incidence of Creutzfeldt-Jakob disease is very low worldwide, sufficient cases are appearing in neurological units to warrant increasing nursing information. In this article epidemiology, etiology, clinical manifestations, and diagnosis, specific nursing diagnoses and suggested areas of intervention with special reference to family involvement are addressed. PMID- 2976789 TI - The effect of transportation between the recovery room and intensive care unit on postoperative acoustic tumor patients. AB - Cardiovascular changes have been noted to occur during movement from the operating room or recovery room (RR) to the intensive care unit (ICU). This study examined blood pressure changes and related variables during transport from the RR to the ICU in 135 postoperative acoustic tumor patients. Although there was no significant change in blood pressure values from the RR to the ICU, patients with pre-existing hypertension and elevated RR blood pressures were noted to have an increased frequency of postoperative complications. In addition, a relationship was noted between administration of halothane and frequency of postoperative complications. Results highlight the need for nurses not only to monitor, but also closely control postoperative blood pressure in this population. PMID- 2976791 TI - A brief neurobehavioral exam useful for early detection of postoperative complications in neurosurgical patients. AB - This article describes data from 11 patients evaluated before and after neurosurgical intervention for treatment of brain tumor, subdural hematoma and hydrocephalus. The Neurobehavioral Cognitive Status Examination (NCSE) was administered pre- and postoperatively. Improvement was documented by the NCSE in 6 of the 11 patients. In the remaining five patients, there was evidence of deterioration in cognitive functioning. After clinical and diagnostic re evaluation by the physician, four patients had repeat operations. Follow-up evaluation after the second operation was useful in determining treatable and non treatable causes of progressive cognitive deterioration. In these patients use of the NCSE may have expedited effective diagnostic evaluation and subsequently improved patient care. Such instruments have clinical utility since they are easily administered, pragmatic, and objectify subtle changes in cognitive functioning which may be early signs of increased intracranial pressure or complications of surgery. PMID- 2976792 TI - Validation of defining characteristics: clinical design. AB - In order to formulate accurate nursing diagnoses, neuroscience nurses must determine clinical validity of defining characteristics. Physiologic, psychologic, behavioral or social signs and symptoms may be studied. This article suggests methods to be used in validating defining characteristics including practical and research design considerations. PMID- 2976793 TI - Two new fluoroquinolones: ciprofloxacin and norfloxacin. PMID- 2976794 TI - Creating a safe environment in the AIDS era. AB - This article discusses concerns related to neuroscience nursing practice in the wake of the AIDS epidemic. Safety issues in the workplace and recommendations for creating a safe environment according to Centers for Disease Control (CDC) guidelines are delineated. Legal considerations from the patient's and nurse's perspectives are also addressed. PMID- 2976795 TI - [The mysteries of docosahexaenoic acid--possible functions of C22 polyenoic fatty acids]. PMID- 2976796 TI - [Prediction of avascular necrosis in the femoral head following fracture dislocation--using the electrochemically generated hydrogen clearance method]. AB - In order to predict necrosis of the femoral head following fracture-dislocation, the blood flow in the femoral heads in twenty-five hips was measured immediately after reduction and three, six, nine and twelve months thereafter. Twenty hips were followed up for more than one year (average 26.3 months) radiographically. Seven out of twenty hips had very low blood flow in the weight bearing areas three months after reduction. Five of seven had very low blood flow six and nine months after reduction and resulted in necrosis which was determined radiographically. The other two of seven recovered the blood flow and have not resulted in necrosis. Thirteen hips with normal blood flow three months after reduction also did not undergo necrosis. It is possible to predict the incidence of necrosis after fracture-dislocation by measuring the blood flow of the femoral head with the electrochemically generated hydrogen clearance method. PMID- 2976797 TI - [Evaluation and medical certificate of disability]. PMID- 2976798 TI - IgE class-specific regulatory factor(s) and Fc epsilon receptors on lymphocytes. AB - The IgE isotype-specific regulatory factor(s) of rodents as well as humans was shown to have an affinity to IgE molecules, suggesting that the factor(s) are Fc epsilon receptors (Fc epsilon R) on lymphocytes or include a fragment of Fc epsilon R. In order to test this possibility, the monoclonal anti-Fc epsilon R antibodies with different epitope specificities were prepared. FACS analysis showed that approximately 50 percent of B cells from normal individuals expressed Fc epsilon R and the augmentation of the expression was observed by the incubation with T cell factors and IgE. However, the Fc epsilon R expression on T cells was not detected even after induction. The result suggests that T cells may secrete Fc epsilon R but not express it on the surface or the IgE-binding factor(s) from T cells may not be antigenically cross-reactive with Fc epsilon R on B cells. PMID- 2976799 TI - Experimental dissection of an IgE class-selective regulatory cascade. PMID- 2976800 TI - The expression of IgE Fc receptors on lymphocytes of allergic patients. AB - Peripheral blood lymphocytes from nonatopic subjects and atopic patients were analyzed for cells expressing Fc receptors for IgE (Fc epsilon R). Nonatopic humans and atopic patients in remission had approximately 1 percent of Fc epsilon R+ peripheral blood lymphocytes. Usually greater than 99 percent of these cells were mIgM+/mIgD+ B cells. However, in approximately 10 percent of nonatopic and atopic subjects a transient increase of Fc epsilon R+ lymphocytes to 3-6 percent was observed in the absence of any disease manifestations and measurable changes in the serum IgE level. At times of increased numbers of peripheral blood Fc epsilon R+ lymphocytes, up to 1 percent Fc epsilon R+ positive cells were detected in isolated T cell preparations. The Fc epsilon R+ T cells reacted with the monoclonal antibody Lyt 3 to the sheep erythrocyte receptor of human T cells but not the anti-T cell antibody OKT3, and fractions also with the monoclonal antibodies OKT8 (cytotoxic and suppressor T cells) and OKM1, which binds to an antigen present on monocytes and a subpopulation of T cells and large granular lymphocytes. No OKT4+ (helper T cells) Fc epsilon R+ cells were detected. The reactivity with monoclonal antibodies to T cell subsets of the Fc epsilon R+ T cells paralleled the reactivity of the IgG Fc receptor positive T cells. In contrast to patients with allergic rhinitis and asthma, patients with severe atopic dermatitis or the Hyper IgE Syndrome always had significantly elevated percentage of Fc epsilon R+ lymphocytes (4-10 percent), which were almost entirely B cells since less than 0.1 percent Fc epsilon R+ T cells were detected in these patients. Atopic dermatitis patients receiving systemic corticosteroid treatment had only 0.2 percent Fc epsilon R+ lymphocytes which was significantly less than the 1 percent of the nonatopic control donors. Attempts to define the function of Fc epsilon R on human B and T lymphocytes have been unsuccessful thus far; however, the increase of Fc epsilon R+ cells associated with atopic disease in man and parasitic infections in rats and mice suggest that Fc epsilon R+ lymphocyte may be involved in the IgE isotype regulation. PMID- 2976801 TI - Regulation of the human IgE antibody response. AB - The frequent association of elevated serum IgE in patients with T cell immunodeficiencies suggest a role for T cells in the regulation of the human IgE antibody response. Unlike the situation with other isotypes the polyclonal B cells activators, pokeweed mitogen and Epstein Barr virus, do not routinely induce IgE synthesis in normal B cells. However, B cells from normal donors will synthesize immunoglobulins of all isotypes (including IgE) when cultured with T cell clones that recognize determinants expressed on the B cells (cognate stimulation). T cells with Fc receptors for IgE can be isolated from patients with hyper IgE syndrome and maintained as long term continuous T cell lines. These cells secrete IgE binding factors which enhance IgE synthesis but not IgG synthesis by preactivated IgE bearing B cells from allergic subjects but not resting B cells from normal donors. IgE binding factors isolated from sera of normal donors selectively suppress IgE synthesis. In contrast, IgE binding factors isolated from sera of patients with hyper IgE syndrome contain IgE potentiating activity as well as IgE suppressor activity. These results suggest that IgE synthesis in man is activated by T cells and isotype specific secretion of this immunoglobulin is modulated by IgE binding factors. PMID- 2976802 TI - IgA binding factors and Fc receptors for IgA: comparative studies between IgA and IgE Fc receptor systems. AB - The expression of Fc receptors (FcR) for IgA (Fc alpha R) as well as for IgE (Fc epsilon R) on T lymphocytes (T cells) is enhanced or up regulated by the corresponding class of immunoglobulins (Ig). The production of class-specific regulatory factors binding to IgA and IgE (IgA binding factor [IgA-BF]; IgE binding factor [IgE-BF]) is also induced by these respective ligands. Murine IgA BFs produced by a T hybridoma T2D4 and concanavalin A-activated spleen cells suppressed the in vitro IgA antibody responses of pokeweed mitogen-stimulated mouse spleen cells class-specifically. Human IgA antibody response was also suppressed by the murine IgA-BF. Similar suppressive IgA-BF is also produced by a human natural killer (NK)-like cell line (YT), which has no rearrangement of the T cell receptor beta-chain gene, indicating that non-T non-B/LGL cells may also be involved in the regulation of the class-specific antibody responses. It appears that, in human as well as murine systems, T- and NK-cells have the capacity to co-express multiple class-specific FcRs and to produce the corresponding immunoglobulin binding factors. While the Fc epsilon R expression is abnormally enhanced in the diseases with hyperimmunoglobulinemia E, disregulation of Fc alpha R is associated with certain human diseases involving the altered IgA regulation. In IgA nephropathy, which is characterized by increased serum IgA level and IgA deposition in the mesangium, there is an enhancement of the expression of Fc alpha R. In contrast, IgA failed to induce Fc alpha R significantly on the lymphocytes from the patients with selective IgA deficiency, indicating that Fc alpha R plays an important role in the IgA regulation in vivo. PMID- 2976803 TI - Fc receptor-bearing T cells and Ig binding factors as class-specific suppressors of polyclonally activated human B cells. AB - By studying the model of polyclonal activation of PBMC from healthy adult humans, selective suppression of the generation of cIgG+ or cIgA+ cells could be achieved by T gamma and T alpha cells activated by Agg-IgG and Agg-IgA, respectively. Furthermore a comparable class-specific suppression was achieved by addition of IgG-BF or IgA-BF released by various cell types including T-enriched PBMC suspensions, B cells or monocytes. The latter effect required the presence of radiosensitive T cells. Whereas T gamma and T alpha cells activated by Agg-Ig inhibited the generation of cIg+ and Ig-secreting cells of the matching class, Ig BFs were shown to act at a transitional stage of B cell maturation by blocking cIg+ generation and/or proliferation, without impairing Ig secretion by fully differentiated plasma cells. Yet another lectin-like factor, termed BMIF, released by FcR- as well as by FcR+ lymphoid or nonlymphoid cells (e.g. polymorphonuclear neutrophils), could block the maturation of cIg+ into Ig secreting plasma cells. Unlike Ig-BF, BMIF was not isotype specific. Cells and lymphokines which control the initial stages of B cell activation and differentiation have been extensively investigated but little is known at present about the regulation of the progression from cIg+B blasts to fully differentiated plasma cells. Sequential determination of cIg+ blasts, plasma cells, PFC, and Ig secretion in polyclonally activated PBMC cultures shows an orderly sequence of appearance and decrease of cells at these different stages, suggesting that up and down regulatory signals control each step. Furthermore the demonstration of suppressor pathways which affect B cell maturation at precise transitional stages provides further indirect evidence towards a sequential regulation of each successive differentiation event. In view of the heterogeneity of FcRs with respect to subclass specificity, affinity, cell type distribution and structure, much remains to be done to elucidate the precise regulatory functions of those molecules in the late stages of B cell maturation. From our studies it would appear that some types of Ig-BF would ensure the recognition of Fc determinants on B cell sIg, but still require T cell, and possibly other factors produced by those cells, to alter B cell maturation. This is in keeping with several models in which isotype specific T cells, but not the Ig-BFs thereof, were shown to regulate B cell differentiation. PMID- 2976804 TI - Isotype-specific recognition and regulation by T cells studied with tumor models. PMID- 2976805 TI - Excessive pronation: a major biomechanical determinant in the development of chondromalacia and pelvic lists. AB - Based on statistical analysis of 97 chronic low back patients, a pelvic list model has been proposed explaining the correlation between leg length discrepancies (LLD) and sciatica. This model suggests that asymmetrical pronation patterns (one pedal arch dropping more than the contralateral pedal arch) initiates a forward downward rotation within the sacroiliac joint. Entrapment of the sciatic nerve between the piriformis muscle and sacrospinous ligament occurs. Clinically, paresis is observed: numbness, weakness and eventually paralysis of the affected limb. Based on this work, the authors have comprised a classification dividing LLD into two main categories: functional and anatomical. Functional LLD have been subdivided into two types of lists: static and dynamic. Treatment based on a combination of chiropractic and podiatric therapy is presented with a 6-month follow-up. Analysis of the success in this tandem approach is very promising. Concurrently, a chondromalacia model is proposed explaining the pathomechanical events associated with oblique tracking patellar syndrome. The authors suggest that excessive pronation is the causative factor directing asynchronous rotation between the shank and femur. This forces the patella out of its normal tracking groove, which, in turn, generates erosion between the inferior margin of the patella and femoral epicondyles. PMID- 2976807 TI - An analytical survey of structural aberrations observed in static radiographic examinations and acute low back cases. PMID- 2976808 TI - Dopamine-derived alkaloids in alcoholism and in Parkinson's and Huntington's diseases. AB - Tetrahydroisoquinoline (TIQ) alkaloids and 1-carboxy TIQ derivatives have been found in human fluids and/or tissues. The possible biosynthetic pathways of salsolinol (Sal), taken as an example of TIQs, are discussed, and the possibility that biosynthesis occurs through a stereospecific enzymatic reaction is considered. In this respect, it is reported that the R enantiomer of Sal predominates in urines of healthy volunteers, whereas the S enantiomer predominates in port wine and possibly in other beverages and foods, suggesting that Sal present in humans could have, at least partially, and endogenous enzymatic origin. TIQs and other dopamine-derived alkaloids are weak MAO inhibitors, the R enantiomer of Sal and salsolidine being more potent than the S form. The changes in monoamine oxidase activity and the nigrostriatal concentrations of dopamine and homovanillic acid in Parkinson's and Huntington's diseases and in alcoholism are reviewed. In these pathological situations, changes in the levels of dopamine-derived alkaloid levels may occur. The possibility that the modifications found might cause or contribute to changes in mental and/or neurophysiological states in these pathological situations is considered. PMID- 2976806 TI - Chiropractic distractive decompression in the treatment of pelvic pain and organic dysfunction in patients with evidence of lower sacral nerve root compression. AB - Chiropractic theory postulates that organic dysfunction could be the result of neurological disorganization secondary to mechanical disorders of the spine. Few studies have documented the efficacy of chiropractic manipulative therapy in treating mechanically induced organic dysfunction. Lower sacral nerve root compression (LSNRC) as the result of lumbar disc lesion has been identified as a cause of pelvic pain and organic dysfunction (PPOD). Ten cases of PPOD with accompanying evidence of LSNRC in patients with low back pain as a result of a clinically established lumbar disc lesion are presented with symptomatology prior to and following treatment with distractive decompressive manipulation. A report of one of the cases is detailed. LSNRC is often overlooked as a cause of PPOD. Recognition of associated symptomatology in patients with evidence of LSNRC and confirmation through pain provocation examination is emphasized. Chiropractic distractive decompression may be effective in treating PPOD in patients with evidence of LSNRC as a result of a clinically established lumbar disc lesion. PMID- 2976809 TI - Genetic mapping of a phosphoglucomutase locus in Aedes togoi. AB - An electrophoretic survey of the phosphoglucomutase (PGM) enzyme was performed using agar gels in 6 strains (3 Japanese strains and 3 strains from Taiwan, Thailand and Canada) of Aedes togoi. The survey revealed at least 3 alleles involved at the Pgm locus among the 6 strains examined. Backcross experiments showed that the Pgm locus was located on the sex chromosome in the following order: Odh (octanol dehydrogenase)--M(sex)--(13.8 map units)--Pgm--(17.0 map units)--h(hooked leg)--s(straw-colored larva). PMID- 2976810 TI - Ca2+-ATPase deficiency in a patient with an exertional muscle pain syndrome. AB - 31P Magnetic resonance spectroscopy studies were carried out in vivo on skeletal muscle of a patient with verapamil-responsive, chronic, progressive post exertional muscle pain. A sister suffered from a similar complaint. The results showed that the muscle: (1) decreased its high energy phosphate content more rapidly than normal during exercise, indicating either increased utilisation or decreased production of ATP; (2) acidified more rapidly than normal during exercise suggesting an increased glycolytic rate; (3) continued in some studies to acidify markedly during the first minute after exercise, indicating that glycolysis remained active into the recovery period; (4) had phosphocreatine and ADP recovery rates consistent with normal rates of oxidative phosphorylation. On the basis of these results, it was proposed that the patient suffers from a defect in Ca2+ handling in the muscle. Subsequently, direct measurement of Ca2+ ATPase activity in the sarcoplasmic reticulum fraction from a muscle biopsy sample showed that the activity of this enzyme was reduced by about 90%. PMID- 2976811 TI - Inhibition of vasopressin secretion during migraine. AB - Six subjects with a history of increased urinary frequency during migraine were studied. In each case there was a marked diuresis and natriuresis within 12 hours of onset of the headache. This was associated with a significant fall in urinary arginine vasopressin (AVP) (p less than 0.01). Migraine may be associated with altered hypothalamic activity resulting in reduced AVP secretion. PMID- 2976812 TI - Absence of antibodies to cardiolipin in patients with Huntington's chorea, Sydenhams chorea and acute rheumatic fever. PMID- 2976814 TI - Physiological responses of rat plantaris motor units to overload induced by surgical removal of its synergists. AB - 1. Rat plantaris muscles were subjected to chronic overload by the surgical removal of the soleus and most of the gastrocnemius muscles. Twelve to 16 wk later whole muscle and motor unit (ventral root dissection technique) contractile properties as well as histochemistry were determined. 2. Motor units were categorized as fast, fatigable (FF), fast, intermediate fatigue-resistant (FI), fast, fatigue-resistant (FR), and slow (S) based on contractile characteristics. Muscle fibers were identified as type I and type II according to myofibrillar ATPase staining. 3. Whole muscles demonstrated increases in wet weight, tetanic force, proportion of type I fibers, and mean cross-sectional areas of both type I and II fibers, as a result of chronic overload. 4. Tetanic tension increased by the same relative magnitude in all motor units whereas twitch tension remained unchanged. A significant change in the proportions of the motor unit types occurred in overloaded muscles, such that the latter contained higher proportions of FF and S units, and lower proportions of FI and FR units, than normal muscles. 5. The fatigue profile of a composite constructed from a summation of motor unit responses revealed that the overloaded plantaris displayed fatigue resistance similar to that of the normal plantaris for a given absolute force output. 6. Glycogen-depleted fibers of hypertrophied single motor units demonstrated uniform myofibrillar ATPase and SDH staining characteristics suggesting that metabolic adaptations among fibers of the same unit were similar after 12-16 wk of overload. 7. The finding that overload caused a uniform increase in the tetanic strength of all motor units, whereas alterations in fatigue resistance varied in degree and direction among unit types, demonstrate that these two properties are not controlled in parallel in this model. The smallest units maintain or even increase their fatigue resistance during the hypertrophic process, whereas high threshold units actually decrease in fatigue resistance. PMID- 2976813 TI - HLA typing and T-cell subpopulations in multiple sclerosis. PMID- 2976815 TI - Plasma, platelet, and aorta fatty acids composition in response to dietary n-6 and n-3 fats supplementation in a rat model of non-insulin-dependent diabetes. AB - Male Sprague-Dawley rats were injected with 90 mg/kg of streptozotocin at 2 days of age. After weaning, they were put on a fat-free diet supplemented with safflower oil (S), a combination of S and linseed oil (L) or a combination of evening primrose oil (E) and L for 8 weeks. Plasma glucose levels and glycosuria were significantly elevated in all 3 groups of diabetic rats in comparison with the corresponding control rats. The percentage of arachidonic acid (20:4n-6) in plasma phospholipids of the S + L and E + L groups was similar to that of the S group and did not differ between control and diabetic rats while adrenic acid (22:4n-6) and docosahexaenoic acid (22:6n-3) changed in proportion to dietary n-3 and n-6 fats content. Arachidonic acid in aorta phospholipids significantly reduced in all 3 groups of diabetic rats as compared to the corresponding control groups. Dihomo-gamma-linolenic acid (20:3n-6) and arachidonic acid in aorta phospholipids increased by the E + L treatment. These results suggest that arachidonic acid in plasma phospholipids is kept constant regardless of the presence of diabetes of non-insulin-dependent type or dietary n-3 and n-6 fats supplementation. In aorta phospholipids, arachidonic acid in diabetic animals reduced and this may be compensated by gamma-linolenic acid supplementation, which leads to increase of dihomo-gamma-linolenic acid and arachidonic acid levels. PMID- 2976816 TI - Drug-induced paralysis (muscle relaxant) therapy in the mechanically ventilated neonate. AB - Drug-induced paralysis in the mechanically ventilated neonate is prescribed primarily to control breathing and, secondarily, to favorably affect underlying pulmonary disease and associated complications. Although the control of breathing can be achieved, it is controversial when pulmonary disease is favorably influenced by paralysis. However, such therapy may lessen the severity, and the incidence of the complications in specific subgroups of infants. In view of significant adverse effects, muscle paralysis should be used judiciously in neonates. PMID- 2976817 TI - Degree of conversion of seven visible light-cured posterior composites. AB - The infrared spectra of seven commercially available posterior composite resins were analysed by ratioing the C = C aliphatic peak to the C = C aromatic peak. The degree of conversion ranged from 43.5 to 73.8%. The result was similar to the range previously obtained from unfilled Bis-GMA based resins and commercial anterior composites. PMID- 2976818 TI - The development of necrotizing enterocolitis following repair of gastroschisis: a surprisingly high incidence. AB - We recently observed the development of necrotizing enterocolitis (NEC) in two consecutive newborn infants after gastroschisis repair. Because this association was unexpected, a retrospective review of our 11-year experience was performed using a multivariant computer analysis. The cohort consisted of 54 newborns with gastroschisis. All infants with omphalocele were excluded. Ten of 54 infants (18.5%) developed NEC as defined by classical clinical findings and pneumatosis intestinalis. Twenty-one distinct episodes of NEC occurred with up to three episodes (mean, 2.1) per patient. Twenty of the 21 episodes were successfully treated nonoperatively. Two infants developed pneumoperitoneum, one of whom underwent laparotomy upon which no perforation or intestinal infarction was found. Eight of the ten patients survived--a survival rate no different than for the remainder of the study group. Neither of the two deaths was attributable to NEC. The NEC was atypical in that no significant relationship was established with known predisposing conditions such as prematurity or low Apgar scores. Suspected risk factors such as time of feeding, type of closure, type of formula, total parenteral nutrition (TPN), and composition of TPN were not statistically related. Significant associations included concurrent TPN associated cholestatic liver disease in nine of the ten infants, antecedent intestinal surgery other than abdominal wall closure in five of the ten infants, and delay in initiation of enteral feedings (greater than 30 days) in eight of ten infants. In addition, the NEC occurred significantly later (range, 32 to 79 days; mean, 52 days) in the clinical course than usual; in fact, three of ten infants were rehospitalized with NEC following discharge. We conclude that a relationship exists between NEC and gastroschisis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976819 TI - Histology of the intestine in human gastroschisis--relationship to intestinal malfunction: dissolution of the "peel" and its ultrastructural characteristics. AB - There are conflicting views on the pathogenesis of the intestinal malfunction seen in infants with gastroschisis. It has been variously ascribed to abnormalities of ganglion cells and smooth muscle elements, intestinal ischemia, and the "peel" which invests the serosa of the intestine. Review of the clinical and experimental literature showed only limited information on the histology of the eviscerated human intestine. In order to add to this data base, and to further investigate the pathogenesis of the intestinal malfunction from a histologic standpoint, we reviewed surgical and autopsy material from our experience with 105 neonates with gastroschisis. Ten specimens were satisfactory for evaluation from a standpoint of tissue integrity. The specific mural components of mucosa, submucosa, muscularis, and ganglion cells were examined and found to be either normal, or to show nonspecific abnormalities that varied from case to case, and were related mostly to intestinal infarction due to compromise of the gut at the site of the gastroschisis defect. In six patients, this progressed to atresia formation. The most consistent abnormalities were found in the serosal layer with its peel. Using special stains, the peel was found to be composed largely of fibrin and collagen. Based on this study, we feel that edema and ischemic changes, though often present, are much less prominent than the peel, as the leading histologic abnormality of the intestine of gastroschisis. Squamous epithelial cells were seen in the peel in four cases, suggesting that the peel had been "appliqued" onto the serosa of the herniated fetal gut. PMID- 2976820 TI - The abdominal wall defect "almost always" lies to the right of the umbilicus. PMID- 2976821 TI - Influence of right atrial stretch and atrial natriuretic factor on rat intestinal fluid content. AB - 1. Studies were made on the effects of right atrial stretch and atrial natriuretic factor (ANF) infusion on fluid movement into the intestinal tract. 2. Stimulation of the atrial volume receptors by inflation of an intracardiac balloon in the conscious, unrestrained rat did not change intestinal fluid content under normovolaemic conditions. 3. When the rat was rendered hypovolaemic by peritoneal dialysis (34% deficit in extracellular fluid volume), right atrial stretch significantly increased intestinal fluid content. Under these conditions, inflation of the balloon restored large intestinal fluid content to the pre dialysis state, i.e. right atrial stretch completely abolished that component of fluid absorption attributable to the extracellular fluid volume deficit. 4. These data suggest that stimulation of the right atrial receptors inhibits intestinal fluid reabsorption but probably does not initiate fluid secretion. 5. There was no evidence that this might be mediated by ANF since rat ANF (twenty-eight residue peptide, Ser-99-Tyr-126), infused for 1 h at doses of 0, 0.05, 0.1 and 0.5 microgram/min, did not cause any detectable changes in the fluid content of the large or small intestine of similarly prepared hypovolaemic rats. PMID- 2976822 TI - The effects of atrial natriuretic peptide and glucagon on proximal glomerulo tubular balance in anaesthetized rats. AB - 1. The renal actions of ANP (average dose 30 ng kg-1 min-1 and glucagon (50 ng kg 1 min-1) were compared using fractional lithium reabsorption as the index of proximal reabsorption in groups of seven rats. Doses were chosen to cause similar increases in glomerular filtration rate (GFR). Time controls were included. 2. Glucagon raised GFR 32% and absolute proximal reabsorption (APR) 26% producing 81% effective proximal glomerulo-tubular balance (GTB) which was not significantly different from the 100% expected for perfect GTB. ANP raised GFR 33% and APR 10% indicating only 30% effective GTB (P less than 0.01). This was a significantly different effect from glucagon (P less than 0.005). 3. Sodium output increased 10-fold with ANP and 3-fold with glucagon. Filtration fraction increased 33% (P less than 0.04) above the pre-treatment value with ANP but was unchanged with glucagon. Plasma renin concentration was suppressed similarly by each hormone (46 and 36%, P less than 0.05, compared with pre-treatment values). 4. Despite a change in peritubular physical factors favouring reabsorption, there was almost complete attenuation of the increase expected in APR with the ANP induced increase in GFR. In contrast, a similar change in GFR with glucagon resulted in an almost parallel increase in APR demonstrating maintenance of proximal GTB. 5. It is concluded that in the anaesthetized rat, ANP but not glucagon profoundly inhibits the increase in proximal reabsorption that normally follows an increase in filtered load. Such an action would contribute to the more potent natriuretic activity of ANP compared with glucagon. PMID- 2976823 TI - Responses to passive movement of receptors in joint, skin and muscle of the human hand. AB - 1. Microneurographic techniques were employed to record unitary activity from afferents associated with digital joints of six conscious human subjects. Of 120 single afferents sampled from the median and ulnar nerves at the wrist, eighteen (15%) were classified as joint afferents; the majority of the sample (72.5%) were of cutaneous origin, and 12.5% were from muscle spindles and tendon organs. 2. Of the eighteen joint afferents six were tonically active in the rest position of the hand. All except two were recruited or accelerated their background discharge during passive joint movement. Three tonically active afferents were responsive to passive movement throughout the physiological range. The majority of the afferents, including the other three tonically active units, responded only towards the limits of joint rotation. 3. As a group, the sample of joint afferents had a limited capacity to signal the direction of joint movement. Nine of the sixteen joint afferents sensitive to movement responded in two axes of angular displacement, and two responded in all three axes. In any one axis of rotation eight afferents were activated in both directions of movement. However, one afferent, associated with the interphalangeal joint of the thumb, responded uni-directionally throughout the physiological range of joint movement and was thereby capable of adequately encoding joint position and movement. 4. Twenty-one of twenty-nine slowly adapting and eleven of eighteen rapidly adapting cutaneous afferents tested were activated by joint movement, but only towards the limits of joint rotation; half of the thirty-two movement-sensitive afferents were bi directionally responsive. Muscle spindle afferents responded to stresses applied to the joint only if the resulting passive movement stretched the parent muscle. 5. It is concluded that human joint afferents possess a very limited capacity to provide kinaesthetic information, and that this is likely to be of significance only when muscle spindle afferents cannot contribute to kinaesthesia. PMID- 2976824 TI - Intracellular pH regulation in resting and contracting segments of rat mesenteric resistance vessels. AB - 1. The pH-sensitive dye 2',7'-bis-(2-carboxyethyl)-5 (and -6)-carboxyfluorescein (BCECF) was used to measure intracellular pH (pHi) in segments of rat resistance vessels (internal diameter about 200 microns) with the vessels mounted in a myograph for simultaneous measurements of isometric contraction. 2. BCECF loaded slowly into the vessels over 1 h and did not affect the maximal contractility of the vessels. There was a loss of dye with time which, however, was very slow when the segments were only excited for 2 s/min, suggesting that the loss was mainly due to dye bleaching with only a very slow leak. 3. The ratio of the emissions (at 540 nm) with excitation at 495 and 450 nm was calibrated in terms of pH using the K+-H+ ionophore nigericin. This calibration gave a pHi value of 7.15 +/- 0.02 (n = 20), suggesting that hydrogen ions are not in electrochemical equilibrium in these vascular smooth muscles which have a membrane potential of about -60 mV. 4. Addition of 10 mM-NH4Cl caused a transient alkalinization and wash-out of 10 mM NH4Cl a transient acidification. Increasing CO2 with maintained bicarbonate caused a rapid acidification followed by an incomplete recovery. Removal of CO2 and bicarbonate (HEPES-buffered solution) with constant extracellular pH caused a transient alkalinization but steady-state pHi was not significantly altered. 5. In bicarbonate-free buffer the Na+-H+ exchange blocker 5-(N-ethyl-N-isopropyl) amiloride (EIPA) and sodium-free conditions caused a slow acidification. In bicarbonate buffer (PSS) EIPA had no detectable effect after 10 min but the anion exchange blocker diisothio-cyanatostilbenedisulphonic acid (DIDS) caused a small acidification over that time course. 6. The rate of recovery after an acid load was about 50% lower in HEPES buffer compared to PSS and it was inhibited by EIPA. In PSS amiloride and EIPA each had a small inhibitory effect on the pH recovery after an acid load. DIDS also inhibited the recovery from an acid load in PSS and this effect was additive to that of EIPA. DIDS and EIPA also had additive inhibitory effects on the 22Na+ influx stimulated by the acid loading, while in HEPES buffer DIDS had no effect on either pH recovery or 22Na+ influx. These results suggest that a Na+-H+ exchange and an influx of bicarbonate coupled to sodium influx are of importance for pHi control in these vessels.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2976825 TI - Instability in human forearm movements studied with feed-back-controlled muscle vibration. AB - 1. Frequency-modulated vibration was applied to the elbow flexor and extensor tendons to produce reflex movements of the forearm in normal subjects. The modulating (command) signal caused equal and opposite deviations from the 40 Hz carrier frequency so that when flexor vibratory frequency increased, extensor frequency decreased, and vice versa. 2. It is argued that the movements resulted largely from the reflex action of muscle spindle primary afferents whose firing frequency had been 'taken over' and modulated by the vibration. 3. Bode plots relating forearm movements to command signal (modulating) frequency showed the transfer function of the Ia afferent-CNS-muscle-load system to have a low-pass filter characteristic. The phase lag of movement on command increased progressively with command signal frequency, exceeding 180 deg at 3-4 Hz. 4. The transduced forearm movements were fed back to provide the command signal to the vibrators (and thus indirectly to the spindle afferents) via a filter mimicking the dynamic responsiveness of muscle spindle primary endings. Our aim was to 'break into' the reflex arc, and re-route it so that we could artificially vary the gain without significantly altering the dynamics of the pathway. 5. Nearly all subjects developed forearm oscillations (tremor) when the gain exceeded a threshold value. Subjects varied widely in their threshold, though for a given subject the threshold remained fairly constant from day to day. The results suggest that reflexly active individuals may not have a large safety margin with respect to forearm instability. 6. The frequency range of the oscillations observed in seven subjects was 3-8 Hz. The frequencies depended upon the level of flexor-extensor co-contraction, and increased from 3 to 5 Hz at 10% co contraction to 5-8 Hz at 100% co-contraction. An analysis of the mechanical impedance of the arm provided estimates of tremor frequencies consistent with these results. 7. These unexpectedly low tremor frequencies led us to propose that it may be erroneous to expect stretch reflexes to contribute to forearm tremor in the range 8-12 Hz (e.g. physiological and 'enhanced' physiological tremors). Rather, their contributions should be sought in the range 3-8 Hz (e.g. pathological tremors such as those of Parkinson's and cerebellar disease). PMID- 2976826 TI - Instability in human forearm movements studied with feed-back-controlled electrical stimulation of muscles. AB - 1. Amplitude-modulated electrical stimulation was applied to the elbow flexors and extensors to produce movements of the forearm in normal subjects. The parameters of the modulating (command) signal were set in isometric trials so as to produce equal and opposite background torques, and equal and supportive torque modulations. 2. Bode plots relating forearm movement to command signal (modulating) frequency showed the muscle-load to have a low-pass characteristic similar to that previously described in the cat, and a slightly larger bandwidth than described previously in man. 3. The transduced forearm signals were fed back to provide the command signal to the stimulators via a filter which mimicked the transfer function of muscle spindle primary endings. In effect this replaced the neural part of the reflex arc with an accessible model, but left the muscle-load effector intact. 4. All six subjects developed forearm oscillations (tremor) when the loop gain exceeded a threshold value. The mean tremor frequency at onset was 4.4 Hz, which was similar to that of the equivalent vibration-evoked tremor (previous paper, Prochazka & Trend, 1988). 5. With the linear spindle model, oscillations tended to grow rapidly in amplitude, and the stimuli became painful. The inclusion of a logarithmic limiting element resulted in stable oscillations, without significant alterations in frequency. This allowed us to study the effect on tremor of including analog delays in the loop, mimicking those associated with peripheral nerve transmission and central reflexes. In one subject, loop delays of 0, 20, 40 and 100 ms resulted in tremor at 4.0, 3.6, 3.0 and 2.1 Hz respectively, as quantified by spectral analysis. 6. By considering separately the phase contributions of the different elements of the reflex arc, including delays, it became clear that muscle-load properties were important in setting the upper limit of tremor frequencies which could conceivably be supported by reflexes. 7. The results support the conclusion of the related vibration study (Prochazka & Trend, 1988), that for moderate levels of background co-contraction, the contribution of stretch reflexes to tremor at the elbow should be sought in the 3-5 Hz range. Exaggerated long-latency reflexes would be expected to reduce these baseline frequencies by 1 or 2 Hz. PMID- 2976827 TI - Further evidence for synaptic actions of muscle spindle secondaries in the middle lumbar segments of the cat spinal cord. AB - 1. The aim of this study has been to investigate the receptor origin of postsynaptic actions evoked by group II muscle afferents in mid-lumbar segments of the cat spinal cord. The experiments tested the hypothesis that the afferents involved were the secondary endings of muscle spindles. 2. Spindle afferents were activated by contractions of intrafusal muscle fibres which were induced by electrical stimulation of fusimotor axons in the distal parts of transected ventral roots by one to three stimuli at 150-500 stimuli/s. A separate series of experiments has shown that such stimuli are effective in activating a considerable proportion of muscle spindle secondaries when contractions of extrafusal muscle fibres are eliminated by differential fatigue of these fibres, provided that several fusimotor axons are stimulated simultaneously. 3. Extracellular field potentials were recorded in the dorsal horn, at such locations where synaptic actions were evoked by electrical stimulation of group II but not group Ia muscle spindle or group Ib tendon organ afferents of pretibial flexors. Effects of activation of spindle afferents following stimulation of fusimotor axons were then compared with effects evoked by electrical stimulation of group II afferents of anterior tibial or extensor digitorum longus nerves and by small stretches of these muscles. 4. Distinct field potentials were evoked by stimulation of ventral root fibres at all locations at which field potentials were obtained from group II afferents stimulated electrically. The latencies of these field potentials were in both cases shorter in the dorsal horn than in the ventral horn. 5. The appearance of these field potentials was not related to contractions of extrafusal muscle fibres and was also observed when these contractions were practically eliminated. Furthermore, their threshold and similar dependence on a potentiating effect of two to three stimuli, as found for single secondaries, allow them to be attributed to secondary endings of muscle spindles. PMID- 2976829 TI - Immunological studies in recurrent spontaneous abortion: effects of immunization of women with paternal mononuclear cells on lymphocytotoxic and mixed lymphocyte reaction blocking antibodies and correlation with sharing of HLA and pregnancy outcome. AB - The occurrence of maternal antipaternal lymphocytotoxic antibody (LCTA), mixed lymphocyte reaction blocking factors (MLRBF) and human leukocyte antigen (HLA) antigen sharing was studied in 115 couples with unexplained repeated spontaneous abortions (RSA). Comparisons were made to the same studies done on 41 couples with explained repeated miscarriages. We found no significant difference between the patient and control group with respect to the percent of couples sharing none, one, or two or more HLA-A,-B, or -DR antigens. Examination of the occurrence of LCTA and MLRBF likewise did not reveal differences between the groups, nor did the occurrence of these antibodies on initial testing correlate with HLA disparity between couples. Women with three or more spontaneous abortions were immunized with paternal mononuclear cells (MNC) if they met at least two of the following three criteria: they shared two or more HLA antigens; their serum was negative for paternal MNC-directed LCTA; their serum did not contain maternal versus paternal MLR blocking factors. Complete HLA, LCTA and MLRBF data pre- and post-treatment are available on 60 women. Sixty-three percent of women converted to LCTA positive 6 +/- 1 weeks after immunization, and 35% of women converted from negative to positive for MLR blocking after immunization. Fifty-eight women who had all three tests done prior to immunization became pregnant after immunization. Only 50% of this selected group have experienced successful pregnancy as judged by delivery of a live-born infant. In the patients presented, successful pregnancy outcome did not correlate with HLA antigen disparity, but successful patients were more likely than aborters to have either LCTA or MLRBF prior to pregnancy (28 vs. 7%). Post-immunization conversion to LCTA positive was more prevalent in the women who aborted after immunization (74%) compared to those who had successful pregnancy (48%) while MLR blocking antibody conversion from negative to positive was the same in both groups. The data indicate that neither HLA antigen sharing nor conversion to LCTA or MLR blocking positive after paternal WBC immunization are predictors for successful pregnancy outcome. Results so far suggest that conversion to LCTA positive after immunization may have a negative influence on pregnancy outcome. PMID- 2976828 TI - Testicular immunosuppressive protein. AB - Auto-, allo- and xenografts of various endocrine tissues survive for prolonged periods in the testicular interstitium. The reason for transplant survival outside the blood-testis barrier has been obscure. In the present paper we describe a high molecular weight (Mr = 130,000), heat- and pH-labile immunosuppressive factor with an isoelectric point of 6.3-7.3 in extracellular fluid collected from the rat testicular interstitium. The results show that the testicular immunosuppressive agent is not a steroid, but a protein. This testicular immunosuppressive protein may contribute to the immune privilege in the testicular interstitium. PMID- 2976830 TI - Augmentation of cytotoxic activity by mitogens in rheumatic heart disease. AB - Natural killer cell activity and alterations in cytotoxicity after culture with streptococcal blastogen A and phytohemagglutinin (PHA) were examined in patients with inactive rheumatic heart disease (RHD) and control patients. Natural cytotoxic activity of mononuclear cells (MNC) did not differ between RHD and control patients with either peripheral blood or tonsils. In cultured blood MNC the level of cytotoxic activity stimulated by blastogen A was significantly greater in patients with RHD at all effector:target cell ratios. These differences in cytotoxic activity were not observed with cultured tonsillar MNC. In similar experiments with a different group of patients, culture with PHA or blastogen A both produced a significantly greater increase in cytotoxic activity in blood MNC from patients with RHD. The increase was significantly lower with PHA than with blastogen A. The ability of mitogens to differentially augment cytotoxic activity in cells from the blood of patients with RHD implies that a population of cells exists in these patients that could be activated during acute rheumatic fever to play a role in pathogenesis. PMID- 2976832 TI - A disability profile of users of a Belfast leisure centre. PMID- 2976831 TI - Chronic upper limb pain syndrome (repetitive strain injury) in the Australian workforce: a systematic cross sectional rheumatological study of 229 patients. AB - The epidemic of chronic upper limb pain is the most important and controversial issue in industrial rheumatology in Australia today. Two hundred and twenty-nine consecutive patients referred with chronic upper limb pain which had been labelled "repetitive strain injury" or "overuse syndrome" were assessed according to a protocol designed to give insight into the questions: Is the pain genuine or falsely reported, i.e., malingering? If genuine is it due to a physical injury, a pain syndrome, or a mixture of both? Twenty-nine patients fulfilled criteria for specific rheumatological diagnoses (fibrositis 15, rotator cuff syndrome 3, rheumatoid arthritis 3, cervical referred pain 3, lateral epicondylitis 2, de Quervain's tenosynovitis 1, carpal tunnel syndrome 1, and psoriatic monoarthritis 1). In the remaining 200 (mean age 37 years, range 19-58, 91.5% female) many different pain patterns and nonspecific associated symptoms were recorded. Eighty nine percent had greater than or equal to 2 Smythe tender points, 1.5% had 1 tender point, and 9.5% had no tender point. Diffuse pain and greater than 7 tender points is sufficient to diagnose fibrositis, and localized pain and a smaller number of tender points strongly suggests a genuine chronic rheumatic pain syndrome. Stress, personal susceptibility and poor motivation appeared important in some cases. The liberal workers' compensation system, early labeling as repetitive strain injury, and social acceptability appeared important in the development of the epidemic. PMID- 2976833 TI - Back and neck problems in a teaching institution. PMID- 2976834 TI - Regression of electrocardiographic left ventricular hypertrophy following treatment of primary hyperaldosteronism. AB - The electrocardiograms (ECGs) of a series of 34 patients with primary hyperaldosteronism (PHA), 17 treated surgically (group I) nine treated medically (group II) and eight treated with/by both surgery and drug therapy (group III) were analysed to determine whether the treatment of PHA resulted in an improvement in the ECG changes of left ventricular hypertrophy (LVH) and also whether there was a difference in this improvement between medically and surgically treated patients. There was a significant reduction of blood pressure in the group as a whole (186/111 to 141/95 mmHg, P less than 0.001) and within each of the treatment groups (group I 183/108 to 137/98 mmHg, group II 188/112 to 147/93 mmHg, group III 193/115 to 144/92 mmHg). This reduction within each group was of a similar magnitude. There were reductions in both precordial voltages (SV1 + RV5) from 3.68 mV to 2.79 mV (P less than 0.01) and in the number of patients with ECG LVH from 15 to 8 (P less than 0.05). Again the reductions were of similar magnitude in each of the groups: group I 3.78 to 2.77 mV, group II 3.78 to 2.84 mV, group III 3.39 to 2.77 mV. Thus the improvement in blood pressure achieved by the treatment of PHA is accompanied by a reduction in precordial voltages and the number of patients with ECG-LVH. This improvement is independent of the type of treatment used. PMID- 2976835 TI - Plasma levels of atrial natriuretic factor in mild to moderate hypertensives without signs of left ventricular hypertrophy: correlation with the known duration of hypertension. AB - The relationship between plasma atrial natriuretic factor (ANF), blood pressure (BP), age, plasma renin activity (PRA) and urinary sodium excretion was studied in 64 normal subjects (mean age 48.7 +/- 2.1 yrs; BP: 126.5 +/- 1.6/79.5 +/- 0.9 mmHg) and in 104 untreated uncomplicated essential hypertensives (50.8 +/- 1.1 yrs; BP: 164.7 +/- 1.6/105.2 +/- 0.6 mmHg). ANF was measured by radioimmunoassay after extraction on C18 columns. ANF was significantly higher in the hypertensives than in the normal subjects (37.1 +/- 1.2 vs 29.7 +/- 1.5 pg/ml, P less than 0.01). In normals plasma ANF was significantly correlated with age (r = 0.72, P less than 0.001), Na excretion (r = 0.42, P less than 0.001) and PRA (r = -0.71, P less than 0.001) whereas in the hypertensives ANF plasma levels correlated only with systolic (r = 0.46, P less than 0.001) and diastolic (r = 0.51, P less than 0.001) BP. In addition in hypertensive patients, by multivariate linear regression analysis, a significant correlation was found between age, known duration of hypertension and plasma ANF. The partial correlation coefficient between duration of hypertension and plasma ANF was highly significant (r = 0.80, P less than 0.001). These findings suggest that in essential hypertension the level of arterial BP is a main determinant of the ANF plasma values offsetting the ability of other physiological factors to regulate plasma ANF levels. PMID- 2976836 TI - Platelet activation indices and apolipoproteins in hypertensive patients. AB - We have studied the platelet activation indices beta-thromboglobulin (beta-TG and platelet factor 4(PF4), triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL) and apolipoprotein (A1, A2, B, C2, C3, E) profiles of 22 untreated essential hypertensive subjects (WHO stages 1 and 2) and 22 controls, to see if there might be some causal relationship between lipoprotein abnormalities and greater platelet activation. The results showed the patients had both greater platelet activation than the controls, as demonstrated by higher plasma beta-TG levels (P less than 0.01) and lower apolipoprotein A2 levels (P less than 0.05). However there were no significant correlations between the platelet activation indices and the plasma levels of apolipoproteins, lipoproteins or lipids in either group. PMID- 2976837 TI - Resolving arguments about the sanctity of life: a response to Long. AB - Thomas Long has argued that there is an irreconcilable metaphysical difference between the views of those who, like ourselves, believe that on quality-of-life grounds it is sometimes justifiable to end the life of a severely handicapped infant, and those who, like Paul Ramsey, reject this view. Because of this metaphysical difference, Long considers it impossible for our arguments to refute Ramsey's position. We disagree. PMID- 2976838 TI - Suffer the little children some autonomy. AB - There exist layers of disparity between perspectives and perceptions of handicap or disability. Handicap is frequently seen by the able-bodied (and minded), as a condition unrelated to themselves. Learned articles appear in specialist journals, but lost in the relentless tidal wave of words, is the very real personality of that handicapped individual ... not waving, but drowning. PMID- 2976839 TI - Palmoplantar keratoderma, nail dystrophy, and hereditary motor and sensory neuropathy: an autosomal dominant trait. AB - Autosomal dominant inheritance of a syndrome comprising palmoplantar keratoderma, nail dystrophy, and hereditary motor and sensory neuropathy (HMSN) was observed in three generations of one family. Nail dystrophy affected the toe and fingernails; it was present at birth or developed during early childhood. Palmoplantar keratoderma became apparent in later childhood. Each subject with nail dystrophy and keratoderma also had clinical or electrophysiological evidence of axonal neuropathy. PMID- 2976840 TI - Genomic imprinting: a possible mechanism for the parental origin effect in Huntington's chorea. AB - Huntington's disease (HD) is an autosomal dominant condition with almost complete penetrance. The age of onset of the symptoms, however, is variable and depends on the parental origin of the gene. A high proportion of early onset cases inherit the HD gene from their father, whereas a considerable proportion of late onset cases inherit the gene from their mother. Modification of the HD gene by maternally inherited extrachromosomal factors has been invoked to account for the parental origin effect. Recent experimental evidence suggests genomic imprinting as an alternative mechanism, by which the gene itself becomes modified in a different way depending on whether it is passed through the maternal or the paternal germline. This modification may involve methylation of DNA and could result in earlier or higher level expression of the gene when it is transmitted by the father. PMID- 2976841 TI - The development of Alzheimer's disease in Down's syndrome assessed by auditory event-related potentials. AB - Several studies have reported changes in auditory event-related potentials in patients with Alzheimer's type dementia. These include an increase in latency and a reduction in amplitude of the P300 (P3) response, a late positive component generated about 300 ms after an unexpected stimulus. Alzheimer's type dementia is an almost invariable acompaniment of ageing in Down's syndrome. This study was designed to assess the usefulness of the auditory P300 response as a measure of the onset of dementia in Down's subjects, who because of poor language development may be difficult to assess by psychological tests. Auditory event related potentials were recorded from 89 Down's subjects, aged 16-66 years. A control group of 29 mentally retarded subjects with fragile-X syndrome and 83 normal volunteer controls were also tested. Clinical psychological testing found evidence of dementia in 16 Down's subjects and none with fragile-X. Furthermore, in the Down's population but not the fragile-X or control groups, there was a marked increase in P300 latency with age starting around 37 years. In controls, the effect of age on P300 latency became significant some 17 years later around the age of 54 years. The premature effect of age on P300 in Down's syndrome was due to the prolonged P300 latency in the 16 subjects showing signs of dementia. It was confirmed that P300 latency increase reflects the development of Alzheimer's dementia in Down's subjects. PMID- 2976843 TI - Vitamin E and Alzheimer's disease in subjects with Down's syndrome. AB - People with Down's syndrome (DS) are at high risk of developing Alzheimer's disease (AD). The gene coding for superoxide dismutase-/ on chromosome 21 resulting in excess activity of the enzyme with consequent risk of oxidative damage might account for the premature ageing. Vitamin E protects against such damage. Plasma vitamin E levels measured in 12 DS subjects with AD (8.19 +/- 0.77 micrograms/ml) were lower (P less than 0.05) than in 12 DS controls (9.43 +/- 1.57 micrograms/ml). It is suggested that there may be an interaction between risk of AD and the protective action of vitamin E. PMID- 2976842 TI - Auditory P300 response in the assessment of Alzheimer's disease in Down's syndrome: a 2-year follow-up study. AB - Sixty-five subjects with Down's syndrome were followed up and retested 2 years after the initial recording of auditory P300 (P3) event-related potential described in a companion paper (Blackwood et al., 1988). The number of subjects showing clinical evidence for Alzheimer's type dementia had increased by a further 14%. In subjects showing clinical deterioration over a period of 2 years, 78% (7/9) had an increase in P3 latency which was three standard deviations or greater than the group mean. None of the 20 fragile-X group retested showed significant change after 2 years. The results suggest that P3 change may be a sensitive index of the onset of Alzheimer's type dementia in Down's syndrome. PMID- 2976844 TI - The significance of Atlanto-axial instability in Down's syndrome. AB - Atlanto-axial instability has been a cause for concern amongst those with Down's syndrome who take part in 'high risk' sports. The authors recommend radiological screening of people with Down's syndrome to help identify those with unstable and symptomatic Atlanto-axial joints and suggest advice on management. PMID- 2976845 TI - Ventricular myosin of the shrew Crocidura russula, correlation with contractile properties. AB - The present study demonstrates that in the shrew ventricular muscle the speed of tension development and relaxation, as well as twitch duration, are much shorter than in the guinea-pig. It also shows that ventricular myosin of the shrew has a high Ca2+-activated ATPase activity and that it is composed of alpha-type heavy chains. Namely, the native molecule is a V1 variety of myosin. These findings advance our knowledge on an as yet uncharacterized mammalian heart and further demonstrate the correlation between mechanical properties and myosin type in heart muscle. PMID- 2976846 TI - Factors affecting the loss of mitochondrial function during zero-flow ischemia (autolysis) in slow and fast heart-rate hearts. AB - The (uninhibited) mitochondrial ATPase comprises approximately 90% of the total ATP hydrolyzing activity present in quiescent, ischemic canine heart muscle and its inhibition by its natural inhibitor protein plays a pivotal role in the slowing of tissue ATP depletion during ischemia. While dog heart mitochondria contain a full complement of mitochondrial ATPase inhibitor capable of fully down regulating the enzyme activity present in this species, rat heart mitochondria contain a much lower level of inhibitor, sufficient to inhibit the enzyme activity present in this species by only approximately 20%. Moreover, this fractional complement of inhibitor remains largely inoperative in the ischemic rat heart. As shown in the present study, one apparent result of the lack of a functional complement of mitochondrial ATPase inhibitor in the rat heart is a more rapid rate of cell ATP depletion during zero-flow ischemia. This in turn results in a more rapidly developed and initially more severe cell acidosis in the ischemic rat heart because ATP hydrolysis produces protons. Finally, and consistent with earlier studies by us, the more rapid ATP depletion together with the more severe acidosis appears to result in a marked increase in the rate of loss of mitochondrial respiratory function in the ischemic rat heart compared to the ischemic dog heart. Our findings suggest that slow heart-rate hearts which contain in situ functional mitochondrial ATPase inhibitor, possess an effective mechanism for sparing cell ATP stores during early ischemia, whereas fast heart rate hearts which lack in situ mitochondrial ATPase inhibitor function, possess a less effective ATP sparing mechanism. PMID- 2976848 TI - A historical perspective of current new attachment procedures. PMID- 2976847 TI - Demineralized bone matrix in treatment of periodontal defects. A review of the literature. PMID- 2976849 TI - The furcation problem: etiology, diagnosis, therapy, and prognosis. PMID- 2976850 TI - [Acute megakaryoblastic leukemia in Down syndrome with 3 q-, dominated by tumor formation and myelofibrosis]. PMID- 2976851 TI - [The effect of percutaneous transluminal angioplasty in renovascular hypertension and the renin-angiotensin system]. PMID- 2976852 TI - [My unborn child]. PMID- 2976853 TI - [Isolated mitral valve insufficiency in comparison with mitral-tricuspid insufficiency: various mechanisms of compensating for the defect and functional status of the myocardium]. AB - A combined analysis of ventricular contractility and intracardiac hemodynamic compensatory mechanisms was carried out, on the basis of angiocardiographic findings, in 37 patients with rheumatic mitral incompetence. Atrial fibrillation aggravates essentially the defect's hemodynamics, while added tricuspid incompetence is accompanied by a certain off-loading in the lesser circulation network. A grossly perversed phasic structure of intramyocardial stress was noted, apparently being a compensatory mechanism. Reduced specific coronary flow and diastolic perfusion gradient in intact coronary arteries are shown to be causes of clinical angina. PMID- 2976854 TI - [Effect of hypertrophy of the left ventricle on myocardial perfusion in patients with arterial hypertension]. AB - A study of 32 patients with arterial hypertension using echocardiography, electrocardiography and myocardial 99mTc-pyrophosphate scintigraphy, demonstrated that developing left-ventricular hypertrophy may interfere with myocardial perfusion. Regression of left-ventricular hypertrophy and recovery of myocardial perfusion is shown to be possible under the effect of antihypertensive treatment. PMID- 2976855 TI - [Umbilical hernia with spontaneous rupture of the hernial sac and skin in ascites]. PMID- 2976856 TI - [Controlled laparostomy in the surgical treatment of peritonitis]. PMID- 2976857 TI - [Selection of the method of plastic surgery of postoperative ventral hernia]. PMID- 2976859 TI - [Structural and functional status of the myocardium in patients with hypertension after long-term use of albetol]. PMID- 2976858 TI - Reduction of Lp(a) by different methods of plasma exchange. AB - Lipoprotein(a) has been shown to be an independent risk factor for atherosclerosis. The effect of different forms of plasmapheresis therapy on the removal of Lp(a) is examined. Plasmapheresis is successfully administered in a small number of hypercholesteremic patients who fail to respond to conventional therapy. Comparison of four different methods of plasma exchange (albumin substitution, anti-apoB antibody column, LDL precipitation, filtration) reveals significant differences in effectiveness in the ability to lower plasma Lp(a): plasma exchange with albumin und LDL precipitation seem to be the most effective, plasma filtration the least. PMID- 2976860 TI - [Algodystrophy of skeletal muscles]. PMID- 2976861 TI - Skin test survey in castor bean allergic working population in eastern Sudan, with frequency response of first dilutions giving skin reactions. PMID- 2976862 TI - Fringe mode transmittance laser Doppler microscope anemometer: its adaptation for measurement in the microcirculation. AB - Blood flow analysis in the microcirculation requires accurate measurement of velocity, volume flow and shear-rate versus shear-stress relationships. The resolution of most anemometers is too limited to obtain useful measurements, especially near the blood vessel wall and at branches and bifurcations. To make such measurements possible with a noninvasive, high resolution, accurate technique, we have developed a fringe mode, transmittance laser Doppler microscope anemometer (LDMA). This system has an intrinsically high spatial resolution (10 x 12 microns), and does not require a high concentration (10(6)/cm3) of scatters or red blood cells (RBC) as in our application. Preliminary measurements of water flow in a rectangular channel were conducted to ascertain the reliability and accuracy of velocity measurements using the LDMA. Velocity profiles were then measured by the LDMA system in arterioles 38-135 microns in diameter, in the transparent, everted cheek pouch of the anaesthetized hamster. The extremely high resolution of the optical system, and the ultra-fine traversing mechanism of the microscope stage, made velocity readings larger than 0.02 mm/s with accuracy and reproducibility better than 1%, possible near the wall to within 7-10 microns. PMID- 2976863 TI - High quality photoplethysmograph signals from a laser Doppler flowmeter: preliminary studies of two simultaneous outputs from the finger. AB - An improvement in the quality of a photoplethysmography signal derived from a laser Doppler flowmeter probe has been achieved by incorporating an auxiliary fibre in the probe head. This fibre is positioned at an optimum distance from the laser light transmitting fibre and overcomes the problem of high frequency signals which mask the detailed features of the photoplethysmograph pulse when a small fibre separation is used. These two signals may be recorded simultaneously from the same site and a correlation between the Doppler output and the photoplethysmograph amplitude has been demonstrated in the finger. The amplitude is shown to be affected by the relative position of the point of measurement with respect to the heart, a factor which does not appear to influence significantly the Doppler output. PMID- 2976864 TI - A week in the life of Mary: the impact of microtechnology on a severely handicapped person. AB - Microtechnology has an important role to play in maximizing the independence of severely handicapped people, especially those with few reliable voluntary movements. Such movements must be harnessed efficiently to provide control over all aspects of life including communication, mobility and the immediate environment. To highlight the role of microtechnology in the day to day life of a severely handicapped person living at home, a case study is presented. The systems Mary uses for communication and environmental control are described and their limitations discussed. Adaptive man-machine interfaces are proposed and foreseeable developments in the fields of speech technology and robotics are considered. Successful introduction of high technology devices requires a full appreciation of medical and social factors as well as those relating to the technology. This is achieved through close partnership between the clinical engineer and the rehabilitation consultant to the benefit of the severely handicapped person. PMID- 2976865 TI - Nonsurgical management of peripheral vascular disease: state of the art. PMID- 2976866 TI - Limitations in the measurement of urine ethanol in clinical trials to monitor ethanol consumption. AB - Self-report of daily alcohol consumption has been used as the dependent variable in clinical trials to assess the effects of two serotonin uptake inhibitory drugs, zimelidine and citalopram. The validity of the dairy data was established by correlating concentration of ethanol in daily urine samples with number of reported standard alcoholic drinks (r = 0.62 for the 934 subject-days in the zimelidine study; r = 0.54 for the 3,103 subject-days in the citalopram study, both p less than 0.0001). The effects of factors other than inaccurate reporting, such as the range of values for reported daily drinks and subjects' drinking patterns, on the correlation coefficients for all subject-days and for individual subjects in the citalopram study are discussed. Sampling 50% or fewer of the 84 days of the citalopram study for each subject is economically advantageous and did not significantly change the values of the correlation coefficients or the rank positions of subjects but did increase the 95% confidence intervals for the correlation coefficients, indicating less certainty about the actual correlation coefficients and, therefore, the accuracy of a subject's self-report. The ability of urine ethanol concentration to validate objectively diaries of alcohol consumption is limited by factors that must be considered but are likely to be out of the investigator's control. PMID- 2976867 TI - [Pre-Sezary erythroderma developing into Sezary syndrome: apropos of 2 observations]. AB - Two cases of pre-Sezary erythroderma of several years of evolution that evolved into Sezary syndrome are presented. The role of persistent antigenic stimulation in the development of T-cell lymphoproliferative disease in discussed. We also review the different therapeutic approach to Sezary syndrome. PMID- 2976868 TI - [Photodynamic destruction of in vitro cultivated squamous cell carcinoma cells of the head and neck area]. AB - Hematoporphyrin derivatives are retained in higher concentrations in proliferating tumor tissues than in normal tissues causing a defined photosensitization. Laser light irradiation of photosensitized cells results in photobiological, non-thermal cell destruction. These photodynamic reactions were analyzed with in vitro cultured squamous cell carcinoma lines of the head and neck. As a light source we used an argon-ion-pumped-dye laser (630 nm wavelength), and for photosensitization we employed a mixture of dihematoporphyrinester and -ether (DHE). The photodynamic reaction on squamous carcinoma cells was proportional to the DHE concentration, the DHE incubation time, and the light exposure. Maximal tumor cell destruction in comparison to DHE free controls was observed with DHE concentrations of 5-10 micrograms/ml medium and a light exposure of 2-4 Joule/cm2. Based on these dose effect relations for DHE sensitized squamous carcinoma cells, photodynamic therapy for destruction of head and neck cancer can be applied in animal tumor models. PMID- 2976869 TI - Hemophilus influenzae type B vaccine. PMID- 2976870 TI - The effects of sufentanil on the hemodynamic and respiratory response to exercise. AB - The effects of the potent opioid, sufentanil, were studied in 11 athletes. Sufentanil was administered intravenously (up to 0.5 microgram.kg-1 over 10 min) to the subjects while they ran at 14 km.hr-1 on a level treadmill. Prior to, and after, the drug infusion, the treadmill was inclined by 6% for 4 min and CO2 was inhaled for 4 min. Two groups were studied: group 1 (six subjects) breathed room air and group 2 (five subjects) breathed O2 enriched air. During level running the ventilation (liters.min-1) of the group 1 subjects was reduced (65.3 +/- 8.6 to 55.9 +/- 4.9, P = 0.09, mean +/- standard error) and PaCO2 (mm Hg) increased from 37.6 +/- 0.7 to 44.0 +/- 0.5 (P less than 0.05). PaO2 (mm Hg) was substantially reduced from 92.0 +/- 2.0 to 70.0 +/- 2.0 (P less than 0.05). In group 2, where hypoxia did not occur, ventilation was reduced from 62.5 +/- 1.5 to 47.6 +/- 1.0 (P less than 0.05). The ventilatory response to the CO2 was shifted to the right but the slope was unchanged by sufentanil. The 6% grade did not cause any significant change in the PaCO2 in either group 1 (0.1 +/- 0.4 prior and 0.8 +/- 0.5 mm Hg increase after sufentanil) or group 2 (1.0 +/- 1.5 prior and 0.4 +/- 0.9 mm Hg increase after sufentanil). The heart rate response was unaffected by sufentanil but the blood pressure increase in response to the 6% grade was blocked with the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976871 TI - Phospholipase A2 and lipocortins, antiphospholipase proteins. PMID- 2976872 TI - Isolation and characterization of genes for cytochrome b6/f complex. PMID- 2976874 TI - Quantification and calibration of flash stimuli in clinical research. AB - Despite the relatively wide use of flash-type stimuli in VEP studies, no universally accepted standards or methods of measurement for these stimuli are in existence. Several methods have been described for the determination of flash intensities, however, the majority of these methods are not sufficiently rigorous, or are based on overly optimistic assumptions on the ideal nature of flash-stimulators. We describe here a simple method for the determination of flash-stimulus energies and absolute peak intensities. Our method does not require specialized optometric instrumentation, and should be readily adaptable to any modern laboratory. PMID- 2976873 TI - An oral contraceptive particularly suitable for women over 35. PMID- 2976876 TI - Pacemaker registration electronic-card. A proposal for a computerised system of storing of the pacemaker registration card. AB - This paper describes an example of a computerised system dedicated to store an Emergency Health Card into a compact and portable memory support such as the CMOS RAM CARD. Details are given from the system used and of the program developed for this task. In particular, patient, pacemaker and lead data are stored by the prototype system. The acquisition layout is similar to that of the European pacemaker registration card but some other sections are added. The possibilities of an introduction of a card system like this on E.E.C. countries is discussed. PMID- 2976875 TI - Impedancimetric bacterial detection: theoretical and experimental aspects. AB - By means of the bipolar impedance technique, we detected bacterial growth in an inoculated broth as its time course absolute impedance. From it, the impedance change relative to sterile medium was obtained, calculating also its time derivative. The repeatability of the derivative curves (they overlapped within a band better than 3.3%) permitted the identification of a double-hump pattern which, in principle, could be accepted as an indicator of the type of bacteria (Escherichia coli). After six experimental series, the growth curves appeared as sensitive to the initial concentration of bacteria and to the culture time preceding inoculation; they were also dependent on the temperature and on the average basal impedance. Temperature showed a greater effect (one order of magnitude) on the lag-phase of the growth curve than on the stationary-phase. This effect occurs because the impedance growth curves tend to get away from the reference offered by the sterile medium. The best working conditions were obtained for an average basal impedance of 510 ohms under well controlled temperature conditions (variations smaller than or equal to 0.20 degrees C) with wire stainless steel electrodes vertically immersed in the culture broth. This impedance technique appears as inexpensive and easy to automatizing for large number of samples. PMID- 2976877 TI - Principles of automatic processing of speech signals and their application in medical technology and for aids for handicapped. AB - Digital processing of speech signals has made great advances in the last few years. This has brought about many new possibilities for man-machine communication in information technology. Automatic recognition and synthesis of speech signals give chances to communicate in a very direct manner with data processing machines. This gives quite good advances in all areas where communication is often done by vocal commands. Medical technology is one example where people normally use spoken commands to get any action or to produce medical documents by voice input. Another and today still much more important area are aids for handicapped people. There are different tasks which may be done using speech recognizers or synthesizers. For paraplegics it will be important to operate their usual aids through vocal commands. For people with communication defects speech recognizers may be useful for the deaf to aid them in participating in speech communication or for training their speech capabilities. For people with speech defects through e.g. spastic paralysis speech aids may help them to produce an intelligible speech signal from written texts, produced by typing on a keyboard. Blind people may use reading machines with speech output to become relatively independent in text reading and text preparation using typewriters. Speech understanding systems may help to make a direct voice dialogue with medical expert systems. On the other hand, advanced research in hearing psychology and the physiology of speech production may help to develop more advanced automatic speech processing. PMID- 2976878 TI - The Nobel Prize and medical progress through technology. PMID- 2976879 TI - Automatic assessment of the interaction between respiration and heart rate variability signal. AB - The present paper introduces an original method of processing heart rate variability (HRV) and respiration signals as detected respectively through chest electrodes and thoracic belt in dogs under different experimental conditions. Signals are processed as time series synchronous with the occurrence of QRS complexes on ECG signal and auto and cross spectra are accordingly calculated. Two particular bands appear mainly of interest on the spectrum of HRV signal: one in correspondence with the respiration rate and another one at a lower frequency value. Values of power at these frequency bands together with coherence and phase between HRV signal and respiration complete the parameters which try to quantify a few aspects of the complex dynamic relationships between the original signals. In particular, controlled respiration in dogs was studied through the connection with an automatic ventilator, as well as the effects of drugs which interact with the neural regulatory systems (i.e. sympathetic and parasympathetic nervous system). Gain and phase relationships between heart rate variability and respiration, obtained with spectral analysis, could be used to provide a better understanding of the neural control mechanisms linking heart rate and respiration in various experimental conditions. The method described in this study is to be used both in physiological and clinical research. PMID- 2976880 TI - Molecular organisation of the quinic acid utilization (QUT) gene cluster in Aspergillus nidulans. AB - The functional integrity of the QUTB gene (encoding quinate dehydrogenase) has been confirmed by transformation of a qutB mutant strain. The DNA sequence of the contiguous genes QUTD (quinate permease), QUTB and QUTG (function unknown) has been determined and analysed, together with that of QUTE (catabolic 3 dehydroquinase). The QUTB sequence shows significant homology with the shikimate dehydrogenase function of the complex AROM locus of Aspergillus nidulans, and with the QA-3 quinate dehydrogenase and QA-1S (repressor) genes of Neurospora crassa. The QUTD gene shows strong homology with the N. crassa QA-Y gene and QUTG with the QA-X gene. QUTD, QUTB, and QUTG, QUTE form two pairs of divergently transcribed genes, and conserved sequence motifs identified in the two common 5' non-coding regions show significant homology with UASGAL and UASQA sequences of the Saccharomyces cerevisiae and N. crassa Gal and QA systems. In addition, conserved 5' sequences homologous to the mammalian CAAT box are noted and a previously unreported conserved 22 nucleotide motif is presented. PMID- 2976881 TI - Alleviation of type I restriction in adenine methylase (dam) mutants of Escherichia coli. AB - The host-controlled EcoK-restriction of unmodified phage lambda.O is alleviated in dam mutants of Escherichia coli by 100- to 300-fold. In addition, the EcoK modification activity is substantially decreased in dam- strains. We show that type I restriction (EcoB, EcoD and EcoK) is detectably alleviated in dam mutants. However, no relief of EcoRI restriction (Type II) occurs in dam- strains and only a slight effect of dam mutation on EcoP1 restriction (Type III) is observed. We interpret the alleviation of the type I restriction in dam- strains to be a consequence of induction of the function which interferes with type I restriction systems. PMID- 2976882 TI - 2-Aminopurine and 5-bromouracil induce alleviation of type I restriction in Escherichia coli: mismatches function as inducing signals? AB - The EcoK restriction of unmodified phage lambda is 1000-fold alleviated in Escherichia coli grown in the presence of base analogs 2-aminopurine (2AP) and 5 bromouracil (5BU). 2AP treatment of bacteria affects specifically the type I restriction systems (EcoA, EcoB, EcoD and EcoK) and does not influence type II (EcoRI) and type III (EcoP1) restriction. 2AP-induced alleviation of restriction occurs in bacteria which are deficient in the SOS response (recA and lexA) and mismatch repair (mutH, mutL and mutS) and can be distinguished from the alleviation of restriction observed in dam- strains. We suggest that mismatches induced by 2AP and 5BU may function as an inducing signal for the alleviation of restriction observed in the presence of base analogs. PMID- 2976883 TI - Ultrastructural aspects of the differentiation of glomerular endothelial cells from the human metanephros. PMID- 2976884 TI - The effect of ethanol upon early development in mice and rats. XIII. The effect of chronic consumption of beer and wine upon preimplantation development in rats. PMID- 2976885 TI - [Embryology of the heart. I. The formation and positioning of the cardiac tube]. PMID- 2976886 TI - Nuclear DNA content in different histologic types of breast carcinoma. PMID- 2976887 TI - Coexistence of two malignant lymphomas in the same patient. A morphological analysis of four cases. PMID- 2976888 TI - Microscopic observations and immunopathologic correlations in bladder carcinomas. PMID- 2976889 TI - Ultrastructural features of the giant-cell tumor of bone. PMID- 2976890 TI - Middle ear infection in suckling and weaned calves. PMID- 2976891 TI - Protective effect of verapamil in experimental myocardial ischaemia: an electron microscopic investigation. PMID- 2976892 TI - Influence of epirubicin and epurox treatment on mouse peritoneal macrophages. PMID- 2976893 TI - Victor Papilian's scientific activity. One hundred years since his birth (1888 1956). PMID- 2976894 TI - Denervation-induced proliferative changes of triads in rabbit skeletal muscle. AB - Protein compositional and functional differences exist between longitudinal and junctional sarcoplasmic reticulum (SR) in relation to Ca transport and to Ca release. In light of this knowledge, we have reinvestigated the effects of denervation on SR of rabbit gastrocnemius, a predominantly fast muscle. Electron microscopy of 2-weeks denervated muscle showed proliferation of transverse tubules (TT), forming junctional contacts with SR terminal cisternae (TC). At coincident periods, the yield of muscle microsomes was increased, and their fractionation by sucrose-density centrifugation demonstrated a relative increase of heavy vesicles. Thin-section electron microscopy of heavy SR from denervated muscle showed an increased number of vesicles containing calsequestrin (CS) as compared with control muscle. Electrophoretic analysis confirmed the relative decrease of Ca-ATPase protein and the striking increase of CS both in total microsomes and in heavy SR vesicles. Calcium loading and Ca-ATPase activity as well as the density of Ca-ATPase protein were decreased to a similar extent (20 30%) in denervated muscle microsomes. Stimulation of Ca-ATPase activity by Ca ionophore A23187 showed that the vesicles were tightly sealed. When probed by competitive ELISA with antibody to SR Ca-ATPase from pure fast muscle, the Ca ATPase of denervated microsomes was found to be highly cross reactive. Cleveland's peptide maps of the Ca-ATPase protein after partial digestion with S. aureus V8 protease also showed no significant change after denervation. Changes in cholesterol content and in the ratio of Mg-ATPase to Ca-ATPase activity of denervated muscle microsomes indicated a 4-fold increase of TT protein, i.e., from about 3% to not more than 12% of total protein, at 2 weeks after denervation. All these changes were totally reversed upon reinnervation of muscle fibers, and the consequent muscle recovery, as obtained by nerve crushing instead of nerve sectioning. From these results, we conclude that denervated adult fast muscle, similarly to immature fast muscle, contains more junctional SR. However, the molecular and catalytic properties of the Ca-ATPase are unaffected by denervation. PMID- 2976895 TI - Metabolic myopathy in canine muscle-type phosphofructokinase deficiency. AB - In vivo 31phosphorus nuclear magnetic resonance spectroscopy (P-NMR) of the anterior tibialis muscle was used to investigate the metabolic myopathy of inherited muscle-type phosphofructokinase (PFK) deficiency in four (homozygous) dogs who had mild exercise intolerance, rare muscle cramps, increased serum creatine kinase activity, but no myoglobinuria. During isometric muscle work induced by indirect electrical stimulation, and subsequent recovery, changes in the ratio of phosphocreatine (PCr) and inorganic phosphates (Pi) were comparable in muscle of PFK-deficient and normal dogs and indicated a large capacity for arobic oxidative phosphorylation in canine muscle. The progressive accumulation of sugar phosphates (PME) during graded exercise clearly demonstrated the glycolytic block in PFK-deficient dogs. During a muscle contracture, induced by acute muscle stimulation, PFK-deficient muscle became completely depleted of PCr and ATP, accumulated large amounts of PME, and recovered very slowly. We conclude that PFK-deficient dogs have a metabolic myopathy that demonstrated some but not all the features recognized in the human disorder. PMID- 2976897 TI - Treatment of severe renovascular hypertension by percutaneous transluminal renal angioplasty in patients with solitary functioning kidney. Effects on blood pressure and renal function. AB - In this study the effects of percutaneous transluminal renal angioplasty on blood pressure and renal function were studied in 9 hypertensive patients with stenosis of the main artery of a solitary functioning kidney. The outcome of the percutaneous dilatation of the renal artery stenosis, and the effects on blood pressure and renal function were evaluated for a mean period of 16.4 +/- 2.13 (SE) months (ranging from 3 to 65 months). A successful dilatation of the renal artery stenosis was shown in all the patients by the aortography performed 1 h after the procedure. At the discharge (7.8 +/- 0.9 days after dilatation), blood pressure was 'cured' in 2 patients and 'improved' in the remaining patients; renal function was improved in all patients who had reduced renal function. At the last follow-up, no restenosis was found in patients who repeated the follow up angiography; blood pressure was 'cured' in 3 patients and 'improved' in 6 patients, and renal function appeared steadly improved. In contrast with other reports, our results demonstrate that percutaneous renal angioplasty is a safe and effective procedure and should be attempted before considering surgical intervention in patients with artery stenosis of a solitary functioning kidney. PMID- 2976896 TI - Cyclopiazonic acid in combination with aflatoxins, zearalenone and ochratoxin A in Indonesian corn. AB - The mycotoxin, cyclopiazonic acid (CPA), was detected at concentrations as high as 9 ppm in 21 of 26 corn samples from a Bogor poultry feedmill. This is the first demonstration of the natural occurrence of CPA in Indonesia. CPA was always accompanied by other mycotoxins, especially aflatoxins, suggesting that the interactive toxicity of these mycotoxins to poultry should be investigated. PMID- 2976898 TI - Effects of 8-OH-DPAT on ultrasonic vocalization and audiogenic immobility reaction in pre-weanling rats. AB - Emotional reactivity in preweanling rats was assessed by observations of two reactions characteristic for two different age periods, respectively. One reaction, ultrasonic vocalization in rat pups separated from mother and littermates, was observed during early postnatal age (10 days). The other reaction, rigid immobility elicited by a sudden sound, was observed at the age of weaning (20 days). Previous studies from this laboratory indicated that the serotonergic system was involved in the control of both of these reactions. In the present study this notion was further tested by investigating the effects on these reactions of the congener of ergot 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), characterized as a centrally-acting, potent and selective agonist of 5-hydroxytryptamine (5-HT). Rat pups, 10 days of age, were treated subcutaneously with 7.5, 15 and 30 micrograms/kg of 8-OH-DPAT and subsequently tested for ultrasonic vocalization. Animals, 20 days old, were treated with 15, 30 and 60 micrograms/kg of 8-OH-DPAT before testing of the immobility reaction. The results showed a dose-dependent decrease of the amount of ultrasonic vocalization and of the duration of the immobility reaction, indicating an anxiolytic-like action of 8-OH-DPAT in both behavioural patterns. A possible explanation for the antagonistic effect of 8-OH-DPAT is that this drug exerts a preferential agonistic effect on presynaptic (auto-) 5-HT receptors. PMID- 2976899 TI - Pupillary response to atrial natriuretic factor in rabbits. AB - This study was designed to evaluate the pupillary and intraocular pressure (IOP) response to exogenously administered atrial natriuretic factor (ANF) in rabbits. Dose-response studies were conducted by administering intracameral (i.c.) ANF (0.1-5 micrograms). The effect of route of administration was evaluated by administering ANF (5 micrograms/kg) intravenous (i.v.) subcutaneous (s.c.) and intraperitoneal (i.p.). In a final study, normal rabbit serum or ANF antiserum were administered i.c. In all studies, pupillary diameter (PD) and IOP were evaluated. Intracameral administration of ANF (0.1-5 micrograms) produced a significant (P less than 0.01) dose-dependent unilateral miosis without affecting IOP. Peripheral (i.v., s.c., i.p.) administration ANF did not affect PD or IOP. Finally, ANF antiserum did not affect PD significantly. These data suggest that ocular, but not circulating ANF may contribute to regulate pupillary function. The mechanism of the miotic response to ANF probably involves interactions with other autonomic neurotransmitters because immunoneutralization of endogenous ocular ANF was without measurable effect. PMID- 2976900 TI - Characterization of beta-endorphin-immunoreactivity in limbic brain structures of rats self-administering heroin or cocaine. AB - The effects of intravenous self-administration of 30 micrograms infusions of either heroin or cocaine, or saline on the concentrations of beta-endorphin immunoreactivity (beta E-IR) in the anterior part of the rat brain limbic system were studied. Self-administration of heroin and cocaine for 5 daily sessions resulted in a marked reduction of the concentrations of beta E-IR in the nucleus accumbens, rostral striatum, septum and hippocampus at the time of the scheduled next session on day 6. In pooled extracts of these regions from rats receiving saline, combined application of high-pressure liquid chromatography (HPLC) fractionation and specific radioimmunoassays revealed the presence of a number of beta E-related peptides co-chromatographing with synthetic non-acetylated and acetylated alpha, beta- and gamma-type endorphins. Similar profiles were found after HPLC fractionation of extracts of these regions from rats self administering heroin and cocaine. Rats self-administering heroin or cocaine, however, showed decreased amounts of all detected forms of beta-endorphin as compared to saline rats. These findings indicate that both self-administration of an opiate that induces psychic as well as physical dependence and of a non-opiate stimulant inducing psychic but not physical dependence, results in a significant decrease of beta E and related peptides in limbic brain regions of the rat. All forms of beta E detected after HPLC were equally affected, suggesting an overall effect of the drugs on peptide turnover. These results suggest that beta E and related peptides may be involved in the neurochemical mechanisms underlying psychic dependence to drugs. PMID- 2976902 TI - [Assessment of the function of language in social medicine expert testimony]. PMID- 2976901 TI - [Current concepts on the diagnosis and therapy of endometriosis]. PMID- 2976903 TI - [Different relations of laws, comprehensive contracts, guidelines and recommendations]. PMID- 2976904 TI - [Social medicine analysis: how sick are rehabilitation applicants? A closer look at expert health insurance assessment]. PMID- 2976906 TI - [Sector related inner city distribution of total cancer mortality in Hamburg]. PMID- 2976905 TI - [Expert assessment and evaluation practice of homeopathic practitioners]. PMID- 2976908 TI - [Professional orientation of former students of the Waldorf school in Helsinki]. PMID- 2976907 TI - [Individual growth of school and kindergarten children--a longitudinal study of height, weight and body mass index percentiles of Basel school and kindergarten children]. PMID- 2976909 TI - [Environmental conditions for the preservation and promotion of health]. PMID- 2976910 TI - [A new fatality caused by dextropropoxyphene]. PMID- 2976911 TI - [Inpatient rehabilitation treatment of malignant neoplasm--an epidemiologic study]. PMID- 2976912 TI - [WHO guidelines for air quality in Europe]. PMID- 2976913 TI - [Organization of obstetrics and prenatal care in Sweden]. PMID- 2976914 TI - [Expert assessment of driving fitness]. PMID- 2976915 TI - [AIDS prevention and the public health service]. PMID- 2976916 TI - ["Reed-rush sewage treatment" from the public health viewpoint]. PMID- 2976917 TI - [Granting special exemptions according to paragraph 4 of the drinking water regulation in plant pesticide contamination]. PMID- 2976918 TI - [Formaldehyde measurements in room air caused by disinfection measures]. PMID- 2976919 TI - [Copper pollution of drinking water]. PMID- 2976920 TI - [Public health officials and politics]. PMID- 2976921 TI - [Allergic symptoms in children and the environment--a pilot project for the assessment of an empirical approach]. PMID- 2976922 TI - [Legal-social observations on prostitution in relation to AIDS and the AIDS catalog]. PMID- 2976923 TI - [Responsibilities of the public health office in monitoring tattooing establishments]. PMID- 2976925 TI - [The public health physician in the emergency service and disaster prevention--a neglected responsibility]. PMID- 2976924 TI - [Results of assessing sterilization and disinfection guidelines in Baden Wurttemberg]. PMID- 2976926 TI - [Unemployment and utilization of the public health system]. PMID- 2976927 TI - [Training of disinfectors--a geographic comparison--emphasis, deficiencies, perspectives]. PMID- 2976928 TI - Inhibition of the activity of glucosyltransferase from Streptococcus mutans by glycyrrhizin. PMID- 2976930 TI - Relative contributions of the rib cage and abdomen during augmented breaths. PMID- 2976929 TI - Diminished radial count is found only postnatally in Down's syndrome. AB - Acinar complexity was assessed using the radial count in 23 patients with Down's syndrome. Late intrauterine growth of the lung is not impaired, and acinar complexity is normal. Thereafter, decreased acinar complexity occurs and is grossly and microscopically apparent by 4 months of age. The gross appearance is characteristic, consisting of a diffuse and uniform porosity of the cut surface of the lung. Microscopically, this appearance is due to the presence of dilated alveoli and alveolar ducts. Children and adults with Down's syndrome have a significantly reduced number of alveoli, which are enlarged. In the majority of cases, the alveoli have a double capillary network that has not been described in any other lung condition. No correlation could be established between the radial count and the presence or absence of double capillary network, hypertensive pulmonary vascular disease, or congenital heart disease. The unique appearance of the lung in Down's syndrome results from failure of the lung to develop properly in the postnatal period, presumably genetically determined. PMID- 2976931 TI - The longitudinal study of the northern Finland birth cohort of 1966. AB - The Northern Finland birth cohort comprises 12,058 live births in 1966-96.3% of all births in the region. The investigation was started during pregnancy and the last follow-up of the total series was at the age of 14 years, when the coverage was still large. Smaller samples of the children and data for the study population from national registers were also examined for the older age groups. The health and development of the children was studied, with special emphasis placed on obtaining reliable incidence figures for neurological handicaps and their correlation with perinatal events. In particular, the correlation between low birthweight and handicapping conditions was documented thoroughly. The indicators predictive of low birthweight among the biological characteristics of the mother and the social conditions of the mother and family, included maternal smoking during pregnancy. The latter was associated not only with adverse perinatal outcome but also with reduction in educational achievement and height among survivors. PMID- 2976932 TI - [Adverse effects of non-steroidal anti-inflammatory drugs]. PMID- 2976933 TI - [Acute non-traumatic rhabdomyolysis]. PMID- 2976934 TI - Degradation of the labelled beta-endorphin is an important factor of direct RIA in the human and rat plasma. AB - Degradation of iodinated beta-endorphin in human and rat plasma was investigated, using different proteinase inhibitors. This degradation affects the radioimmunoassay quality. Additional blank tubes were also prepared during assay procedure to avoid the effect of 125-I-beta-endorphin adsorption on the dextran coated charcoal in the presence and absence of plasma proteins, what avoids underestimation of the beta-endorphin level in plasma samples. Based on the above observations, the sensitive and direct radioimmunoassay of beta-endorphin was developed in human and rat plasma. PMID- 2976935 TI - Scientific opinion on the carcinogenic risk due to topical administration of benzoylperoxide for the treatment of acne vulgaris. PMID- 2976936 TI - Toxicity of immunotherapy with interleukin-2 and lymphokine-activated killer cells. AB - Immunotherapy with IL-2 represents a major breakthrough in the management of renal cell carcinoma and malignant melanoma. At present, the toxicity of most IL 2 regimens is severe and prohibitive for clinicians not intimately familiar with the myriad of side effects associated with its use. The elucidation of the mechanism by which the lymphokine induces tumor regression, the vascular leak syndrome and other side effects will permit IL-2 to be used more safely and effectively. PMID- 2976937 TI - [Dynamics of interleukin 2 production in patients with diffuse toxic goiter under the influence of treatment]. AB - The paper is devoted to the results of a study of IL-2 production by T-helpers in the course of treatment of patients with diffuse toxic goiter (DTG); 30 patients and 30 healthy controls were investigated. The diagnosis of DTG was confirmed by modern methods including radioimmunoassays. A study of the blood level (relative and absolute) of T-lymphocytes in the patients before therapy, during 2 and 12 mos. of thimazol therapy and during remission of disease showed its significant decrease as compared to the controls. IL-2 production in the blood lymphocyte culture supernatant of the DTG patients was assessed by testing on 3H-thymidine labeled ConA-T-blasts. A rise of the stimulation index before therapy and during 2 mos. of therapy, its normalization at the subsequent stage of therapy and the reduction in the patients with remission of thyrotoxicosis were established. A double control of IL-2 production (resulting from therapy and the level of thyroid hormones T3 and T4 decreasing during therapy) was assumed. The results of the study suggested the selectivity of affection of T-lymphocyte subpopulations in DTG and confirmed the preserving of T-helper function. PMID- 2976938 TI - Predicted structure for the calcium-dependent membrane-binding proteins p35, p36, and p32. AB - A new family of proteins (annexins) that bind to membranes at micromolar free Ca2+ has been recognized. Its members include an EGF-receptor kinase substrate (p35), a retroviral tyrosine kinase substrate (p36), the liver protein endonexin (p32) and an electric ray protein, calelectrin. Each protein contains four sequence repeats with a further 2-fold internal homology. Using the predicted secondary structure and pattern of conserved hydrophobic residues in each repeat, we have built a three-dimensional model that is largely isostructural with the known molecular conformation of bovine intestinal calcium-binding protein. The final (energy-refined) model had a core formed from the conserved hydrophobic residues. It differed from ICaBP principally in the length of the two Ca2+ binding loops with only one loop being able to bind. The model suggests a mechanism for interaction of these new Ca2+-binding proteins with phospholipid bilayers. PMID- 2976939 TI - Selenocysteine's mechanism of incorporation and evolution revealed in cDNAs of three glutathione peroxidases. AB - The nonsense codon, UGA, has for the first time recently been shown to encode selenocysteine in two proteins, mouse glutathione peroxidase (GSH-Px) (EC 1.11.1.9) and bacterial formate dehydrogenase. A co-translational rather than post-translational selenium-incorporation mechanism has been implicated. Furthermore, high expression levels of GSH-Px have suggested that suppression of termination is efficient and specific. We have isolated and characterized pituitary, kidney and placenta cDNAs for bovine, human and mouse GSH-Px respectively. It is demonstrated that this novel suppression event occurs in diverse tissues, in at least three mammalian species and at the translational step. Surprisingly, GSH-Px is shown to be extramitochondrially encoded, indicating a cytosolic suppression event rather than one utilizing the mitochondria's well-documented extended codon-reading ability. Sequence analysis reveals that a simple proximal contextual pattern responsible for readthrough does not exist. Analysis of predicted secondary structures of mRNAs, however, has revealed a conformation which may be unique to selenocysteine proteins and may prove useful as a tool for artificial incorporation of selenocysteines. A human intron for GSH-Px from an unspliced mRNA has been isolated whose position indicates an ancient, divergent evolutionary relationship with thioredoxin-S2, rather than an independent convergent one. PMID- 2976940 TI - Primitive rhabdomyosarcoma presenting with diffuse bone marrow involvement: an immunohistochemical and ultrastructural study. AB - We recently identified three cases of primitive rhabdomyosarcoma (PRMS) presenting with diffuse bone marrow infiltration but inconspicuous soft tissue primaries, referred to The Johns Hopkins Hospital (JHH) as acute leukemia. In each case, the diagnosis of rhabdomyosarcoma was established using immunohistochemical staining and electron microscopy. Ultrastructural examination of tumor cells showed a feltwork of thin filaments, discontinuous basal lamina, glycogen, and primitive cell junctions without cell processes or neurosecretory granules. This presentation of PRMS may be more common than recognized, since it can be readily misdiagnosed as a hematopoietic tumor. While positive staining for muscle-specific actin, desmin, myoglobin, or other markers of skeletal muscle differentiation may be diagnostic, negative staining is inconclusive, requiring recognition of the "minimal" ultrastructural findings of primitive rhabdomyosarcomas. PMID- 2976941 TI - Nuclear morphology of follicular center cell-associated T-cells: an immunoultrastructural study of follicular hyperplasia and follicular center cell lymphomas. AB - Although the immunophenotypic nature and distribution of T-cells in normal follicular centers and in follicular center cell lymphomas have been studied in great detail, the morphology of these cells has been largely ignored. Immunoultrastructural studies using UCHT-1, an antibody to the T-cell receptor associated CD3 antigen, were therefore performed on four tonsils with reactive follicular hyperplasia and on four follicular center cell lymphomas. T-cells present in normal follicular centers had a mean maximum nuclear diameter of 4.9 +/- 0.93 microns and were not significantly different from those associated with neoplastic follicular center cells which had a maximum nuclear diameter of 4.9 +/ 1.2 microns. In contrast, the T-cells in normal interfollicular T-zones had a mean maximum nuclear diameter of only 4.4 +/- 0.81 microns which was significantly smaller than either of the previous two groups (P = 0.002 and P = 0.004, respectively). Similarly, T-cells in a relatively uninvolved T-zone in a follicular center cell lymphoma had a mean maximum nuclear diameter of 4.5 +/- 0.78 microns. The nuclear configuration of the follicular center cell-associated and interfollicular T-cells ranged from round to clefted with the greatest number of clefted T-cells being associated with the follicular center cell lymphomas. These data demonstrate that T-cells associated with either normal or neoplastic follicular center cells tend to be larger than interfollicular T-cells. Because of the very variable nuclear configurations of these T-cells, not all cells with clefted nuclei in follicular centers or follicular center cell lymphomas can be assumed to be of B-cell origin. PMID- 2976942 TI - Influence of urapidil on in vitro platelet response to adrenaline and other aggregating agents. AB - The effects of hypotensive drug urapidil on human platelet functions were investigated. Urapidil failed to evidence direct aggregating properties or potentiating effects. Furthermore, drug high concentrations inhibited the platelet response to ADP, PAF, collagen, adrenaline and bovine thrombin, and influenced the platelet release reaction induced by ADP and PAF. Data indicate that urapidil possesses negligible agonistic effects on human platelet alpha 2 adrenoceptors and interferes at high concentrations with the platelet activation, as evidenced for other anti-aggregating compounds. PMID- 2976943 TI - Copulatory performance of penile desensitized male rats following the administration of 8-OH-DPAT. AB - Bilateral transection of the dorsal penile nerve in male rats was followed by reduction of the amount of intromissions in relation to the frequency of mounts and abolishment of the capacity to ejaculate. Treatment with 8-OH-DPAT (0.25 mg.kg-1 -20 min SC) essentially normalized the behavior of the operated animals which showed a normal proportion of mounts to intromissions and which all ejaculated at almost all testing occasions. It was speculated that treatment with 8-OH-DPAT makes central neural mechanisms more responsive to genital stimulation thereby facilitating the development of full penile erection, vaginal insertion and intromission. PMID- 2976944 TI - Evidence for serotonergic modulation of sucrose sham-feeding in the gastric fistulated rat. AB - Both the real and sham intakes of a 5% sucrose solution were reduced by d fenfluramine, quipazine (a 5-HT receptor agonist), and fluoxetine (a selective 5 HT reuptake inhibitor). These data demonstrate a serotonergic inhibitory component in the control of sham-feeding. In support of this, 8-OH-DPAT (a selective 5-HT1A agonist), which inhibits serotonergic activity at small doses via an autoreceptor mechanism, enhanced sucrose sham-feeding and reversed the suppressant effect of d-fenfluramine. Together, the results indicate a serotonergic influence on ingestional responses which is probably closely related to the oropharyngeal control of consumption. PMID- 2976945 TI - Profile of the selective dopamine D-2 receptor agonist N-0437: its effects on palatability- and deprivation-induced feeding, and operant responding for food. AB - N-0437 is a potent and highly selective dopamine D-2 receptor agonist, which has been used in the present series of experiments to investigate its potential anorectic properties. In doses of 0.3-3.0 mg/kg (IP), N-0437 significantly reduced consumption of a sweetened palatable mash in nondeprived mice (minimal effective dose, 0.3 mg/kg) and rats (minimal effective dose, 0.56 mg/kg). Reduction in food intake were also produced in rats by the less potent, but selective, D-2 agonist RU 24213 (effective at 10.0 mg/kg), and by d-amphetamine (1.0 and 3.0 mg/kg). The anorectic effect of N-0437 (1.0 mg/kg) was completely antagonized by the selective D-2 antagonist, YM-09151-2 (0.01 mg/kg). Over a series of 10 injections, N-0437 (1.0 mg/kg) maintained its effect to reduce palatable food intake. In food-deprived rats, N-0437 (0.3-3.0 mg/kg, IP) also reduced consumption of standard laboratory food, and dose-dependently reduced operant responding for food under a FR8 schedule of reinforcement. The results of the experiments are discussed in terms of a possible direct effect to reduce feeding responses resulting from stimulation of postsynaptic dopamine D-2 receptors. PMID- 2976947 TI - Platelet 3H-paroxetine binding in depressed patients. AB - Studies using 3H-imipramine binding in platelets as a means of exploring the serotonin transport system in depressed patients have produced inconsistent findings. For this reason, we studied 3H-paroxetine binding in platelets, because we believed it might provide a more reliable ligand for studying serotonin transport. Subjects were 23 depressive inpatients and 23 normal controls. No differences in the maximal number of binding sites (Bmax) or in the dissociation constant (Kd) were found between depressives and controls. PMID- 2976946 TI - [Atlanto-axial dislocation in Down syndrome and os odontoieum. Case report and review of the literature]. AB - A case is reported of the rare coincidence of mongolism and os odontoideum in an eleven-year-old child, leading to compression myelopathy in consequence of atlantoaxial dislocation, and an overview of literature to the present time is given. Recommendations on diagnostic and prophylactic measures are offered designed to counteract as early as possible the spinal complications of atlantoaxial instability. PMID- 2976948 TI - WAIS--R factor structure in a vocational rehabilitation sample: additional support for a third factor in special populations. PMID- 2976949 TI - Premenstrual symptom changes and plasma beta-endorphin/beta-lipotropin throughout the menstrual cycle. AB - The beta-endorphin (BE) hypothesis of premenstrual symptomatology was assessed in 15 healthy, drug-free normally cycling women. Cycle-phase assignment was aided by measurement of basal body temperature and plasma progesterone. For the entire group, peripheral plasma BE concentrations by radioimmunoassay (with substantial beta-lipotropin (BL) cross-reactivity) were generally unvarying across the menstrual cycle. High and low symptom subgroups were defined by retrospective and prospective daily self-reports of premenstrual changes. For the cycle studied, moderate to extreme symptom severity was defined according to premenstrual increases in impaired concentration, water retention and negative affect. No between-groups difference was observed for any cycle-phase for absolute values of beta-endorphin (BE/BL). When difference scores were examined, there was a positive time-lag correlation between follicular-to-periovulatory changes in BE/BL and the subsequent luteal increase in impaired concentration over the periovulatory baseline (rs = 0.82); considering the number of comparisons, however, the correlation fails to reach significance. The salience of various types of stressors on mood was assessed; there was a trend for "task-demands" to account for a greater percentage of negative attributions premenstrually compared to post-menstrually, for the entire group. The relative inconsistency in plasma BE/BL from cycle to cycle may be related in part, to the lack of stability in certain cycle characteristics from month to month. Future studies of BE/BL across the menstrual cycle may benefit from a demanding task in a provocative stress paradigm. PMID- 2976951 TI - Centrally injected atrial natriuretic factor inhibits angiotensin- and osmotically-induced drinking in pigs. AB - Atrial natriuretic factor 5-28 (atriopeptin III, APIII; 10 micrograms) injected into the cerebral ventricles of young pigs 5 min before injection of either angiotensin II (AII, 300 ng) or hypertonic NaCl (0.74 M) attenuated the subsequent drinking responses to these dipsogens although the latency to the response was not altered. The inhibition lasted for up to 4 days in the case of AII-induced drinking but normal daily water intake was not disrupted. These findings suggest that in the pig APIII may play a role in the central regulation of drinking in response to AII and that it may have a long biological half-life. PMID- 2976950 TI - Major depressive disorder predicts cardiac events in patients with coronary artery disease. AB - Fifty-two patients undergoing cardiac catheterization and subsequently found to have significant coronary artery disease (CAD) were given structured psychiatric interviews before catheterization. Nine of these patients met criteria for major depressive disorder. All 52 patients were contacted 12 months after catheterization, and the occurrence of myocardial infarction, angioplasty, coronary bypass surgery and death was determined. Results of the study show that major depressive disorder was the best predictor of these major cardiac events during the 12 months following catheterization. The predictive effect was independent of the severity of CAD, left ventricular ejection fraction, and the presence of smoking. Furthermore, with the exception of smoking, there were no statistically significant differences between those patients with major depressive disorder and the remaining patients on any variable studied. The possible mechanisms relating major depressive disorder to subsequent cardiac events are discussed. It is concluded that major depressive disorder is an important independent risk factor for the occurrence of major cardiac events in patients with CAD. PMID- 2976952 TI - The effects of diphosphonates on alveolar bone loss. PMID- 2976953 TI - [Contact dermatitis in Krosno]. PMID- 2976954 TI - [A new type of contact allergens: airborne allergens]. PMID- 2976955 TI - [The role of metals in the development of allergy in workers in the electrotechnical industry]. PMID- 2976956 TI - [13-cis-retinoic acid in the treatment of acne conglobata et phlegmonosa in the light of our studies]. PMID- 2976957 TI - Atrial natriuretic peptide inhibits water and sodium intake in rabbits. AB - The effect of atrial natriuretic peptide (ANP) on water and sodium intake was investigated in wild rabbits, a species which does not drink water following i.c.v. or i.v. administration of angiotensin II but develops sodium appetite following i.c.v. infusion of angiotensin II. ANP was given during or after depletion of extracellular fluid volume: hemorrhage, fluid deprivation and administration of furosemide. Systemically administered ANP reduced the water, but not the sodium intake of wild rabbits. I.c.v. administration of ANP inhibited both water and sodium intake. The suppression of thirst following both i.v. and i.c.v. administration of ANP indicates that inhibition of the effect of angiotensin II is not the exclusive mechanism and the circumventricular organs are probably not the exclusive sites of action for ANP. The inhibition of sodium appetite in wild rabbits was consistent with earlier proposals that ANP acts through the inhibition of the effects of angiotensin II. Reduction of food intake coincident with administration of ANP was also noted, but dose-dependent decrease was not observed. PMID- 2976959 TI - [Clinical contributions to the always open file of so-called essential hematuria]. PMID- 2976958 TI - Effect of central hypertonic stimulation on plasma atrial natriuretic peptide and oxytocin in conscious rats. AB - The effect of the intracerebroventricular (i.c.v.) injection of hypertonic sodium chloride on plasma atrial natriuretic peptide (ANP) and oxytocin (OT) was evaluated in conscious freely moving rats. A hypertonic or isotonic NaCl solution was injected into the third ventricle. Blood pressure and heart rate were monitored and blood samples were collected. I.c.v. injection of the hypertonic solution resulted in a significant increase in mean arterial pressure (105.3 +/- 2.9 mmHg at time 0 to 124.2 +/- 4.4 mmHg at 5 min, P less than 0.01) and heart rate (350.0 +/- 25.0 bpm at time 0 to 420.8 +/- 13.6 bpm at 20 min, P less than 0.01). Plasma OT increased 4-fold over the basal values 5 min after the injection (4.5 +/- 1.1 to 20.1 +/- 3.2 pg/ml, P less than 0.01), while there was no significant change in plasma ANP (37.3 +/- 9.1 to 46.6 +/- 12.6 pg/ml, n.s.). The control injection produced no significant changes in any parameters. These results show that hemodynamic changes are not necessarily associated with alterations in plasma ANP. Furthermore they suggest that central osmoreceptors are not involved in the control of ANP secretion. PMID- 2976960 TI - [Primary evolutive papillo-odditis An anatomico-clinical and therapeutic study]. PMID- 2976961 TI - [Autografting of the traumatized spleen]. PMID- 2976962 TI - [Autotransplantation of the 2d toe in the reconstruction of the index finger. The microsurgical technic]. PMID- 2976963 TI - [2 benign proliferative diseases of the stomach with a major surgical solution: hypertrophic gastropathy with giant folds and diffuse gastric polyposis]. PMID- 2976964 TI - [Jejunocutaneous fistula following the use of a polyester net for treating eventration]. PMID- 2976965 TI - [Hemodialysis via a venous catheter in the femoral or subclavian vein in the urology patient with kidney failure]. PMID- 2976966 TI - [Antithrombotic therapy in cardiac disease: approach for the selection of pharmacologic agents]. PMID- 2976968 TI - [Residual ischemia after myocardial infarct (II). Treatment]. PMID- 2976967 TI - [Percutaneous aortic valvuloplasty in senile calcified aortic stenosis in 15 patients. Immediate results and 12-month follow-up]. PMID- 2976969 TI - [Insufficiency of the left ventricle in elderly patients with normal systolic function. Is ischemia a determinant of the disease?]. PMID- 2976970 TI - [Immediate coronary angioplasty in patients with acute infarct of myocardium treated with tissue plasminogen activator: evaluation with cardiac perfusion gammagraphy in exertion]. PMID- 2976972 TI - [Rehabilitation overseas. Integration of handicapped students at American community colleges]. PMID- 2976971 TI - [The personal computer as an aid for the handicapped and as a support measure in the rehabilitation of handicapped patients]. AB - Our working world cannot be pictured anymore without computers of all kinds and sizes - which renders them equally relevant to the field of rehabilitation to disabled people. What is less known and accepted is the fact that the computer constitutes a technical device excellently suited for a multitude of purposes in the day-to-day life of disabled persons. Commercially available devices and programmes frequently can be used directly, and adaptations of earlier developments can be transferred with only minor modification. The present contribution is intended to give an idea of currently available, practical experience-based possibilities for highly efficient, and at the same time economical, provision of technical aids making use of personal computers. PMID- 2976973 TI - Mechanical properties of isolated fetal miniature pig lungs after substitution with fluorocarbons. AB - In our present study we tried to inflate and stabilize isolated immature lungs of fetal minipigs in gestation age of 95 days (= 85% of total normal gestation period) with different fluorocarbons. Based on our previous experiences, the immature lungs at day 95 are almost non inflatable with air. For our experiments we used fluorocarbon 43 (FC-43) with a surface tension of 16 mN/m and fluorocarbon 72 (FC-72) with a surface tension of 12 mN/m. Eighteen fetal immature lungs were used. In group 1 the lungs were rinsed with FC-43; in group 2 the rinse solution was FC-72, and in group 3 the lungs were untreated. After removing the fluorocarbon, in the case of groups 1 and 2, the lungs were artificially ventilated. Pressure-volume (p-v) curves were registered in the beginning (immediately after FC lavage), after 10 and 20 min of artificial ventilation. Airway opening pressure (pi) and weight-specific end-inspiratory lung compliance (ci) were investigated. Statistically significant differences in weight-specific end-inspiratory compliance were found between FC groups and untreated group 3, but no stabilization could be seen during the investigation period of 20 min. No statistically significant improvement in weight-specific end inspiratory compliance was observed between group 1 and 2, although the compliances of group 2 with FC-72 were better than those of group 1 with FC-43 in three p-v diagrams registered in the beginning and after 10 and 20 min of artificial ventilation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976975 TI - [A case of valvular disease with giant right atrium and partial atrial standstill]. PMID- 2976974 TI - Plasma concentrations of atrial natriuretic factor in acute left ventricular failure in the dog. AB - Acute ischemic left ventricular failure was induced in anesthetized dogs by repeated coronary embolization with 50 microns microspheres. Plasma concentrations of atrial natriuretic factor (ANF) were measured by radioimmunoassay of arterial and venous samples before and after failure induction. Heart failure was accompanied by a doubling of arterial ANF concentration, whereas there were only insignificant changes on the venous side. The increase in arterial ANF correlated significantly to the increase in left ventricular end-diastolic pressure, but not to the increase in right atrial pressure. Measurements of pericardial pressure indicated that a pericardial constraint acted to reduce atrial distension and thereby cardiac ANF release during failure. PMID- 2976976 TI - [Adverse reactions to anti-hepatitis B vaccine in hospital personnel: results end experiences]. AB - It has been studied, by inquiry, the adverse reactions in the hospital personnel vaccinated against Hepatitis B with 3 doses of 20 mcg of the HB-VAX (MSD) vaccine. The 45% of the inquired people referred some kind of the side effects, more frequently after the first vaccinal dose. The local reactions incidency was larger than the general ones, but without any significant differences. The local pain and the asthenia, general malaise and myalgia were the more outstanding symptoms in both cases. The average duration of the adverse reactions was two days, not appearing any disorders in the laboral activity of the vaccinated. In our experience, the anti-Hepatitis B vaccine employed didn't offer superior risk to the observed with antiviral vaccines employed to prevent other diseases. PMID- 2976978 TI - IgG receptors on eosinophils--using immune scanning electron microscopy. AB - Using carboxylate modified latex particles covalently conjugated with anti-IgG, IgG receptors on eosinophils were examined with scanning electron microscopy (SEM). After block and inhibition tests, significant number of latex particles were confirmed to bound on the surface of eosinophils. Earlier reports described that density of eosinophils decreased in hypereosinophilic patients and the heterogeneity of eosinophils came into focus. Our experiment revealed that eosinophils of hypereosinophilic patients had more IgG receptors than those of normal volunteers. This difference might be due to the heterogeneity of eosinophils. PMID- 2976977 TI - Increased atrial natriuretic peptide in an early stage of chronic glomerulonephritis. AB - Atrial natriuretic peptide (ANP), angiotensin II (AII), aldosterone (Aldo), arginine vasopressin (AVP) in plasma, urinary excretion of prostaglandin E2 (PGE2) and urinary sodium excretion rate (UNaV) were determined in 11 normotensive patients with chronic glomerulonephritis and a normal glomerular filtration rate (GFR) and in 14 healthy control subjects before, during and after intravenous infusion of a 2.5% sodium chloride solution. During basal conditions ANP was increased in patients compared with controls (9.8 pmol/l (median) versus 7.2 pmol/l, p less than 0.01). After sodium infusion ANP was unchanged in the patients but significantly increased in the controls. AII, Aldo, AVP in plasma and urinary PGE2 excretion were the same in patients and controls. The urinary sodium excretion rate was significantly increased in patients compared with controls during sodium infusion (p less than 0.05). No correlations were found between ANP and UNaV, AII or Aldo in either patients or controls. The relationship between serum osmolality (Sosm) and AVP was normal in the patients. It can be concluded that in normotensive patients with chronic glomerulonephritis and normal GFR, ANP is increased during basal conditions and the response to acute volume expansion may be blunted. The renin-angiotensin system, the osmoregulatory system and urinary PGE2 excretion are normal and respond in a normal way to volume expansion. It is suggested that the increased level of ANP can be viewed as a compensatory phenomenon to an abnormal sodium or volume homeostasis in the early stages of chronic glomerulonephritis. PMID- 2976979 TI - [The clinical significance of ECG exercise testing at hospital discharge of patients with acute myocardial infarction]. AB - 330 patients were admitted to the intensive care unit of our hospital from January 1985 to December 1986. 141 of the 150 patients aged below 65 years underwent symptom-limited maximal exercise testing on a bicycle ergometer before discharge. No severe complications were noted. 61 of those tested showed a pathological reaction such as angina pectoris, ST-segment depression or ventricular ectopic activity (couplets, ventricular tachycardia). Coronary angiography was performed in 33 patients (23.4%). The outcome of this examination led to coronary bypass surgery in 15 cases and to PTCA in 2. For 9 of these 17 patients exercise testing soon after myocardial infarction was the essential investigation that led to angiography and surgery. Symptom-limited exercise testing before discharge is a safe method of distinguishing patients at risk, requiring further investigations for possible coronary bypass surgery, from those that do not need additional work-up. We conclude that every post-myocardial infarction patient should be exercise-tested before leaving hospital. PMID- 2976980 TI - [Determining the optimal period of temporary disability in various diseases of the internal organs]. PMID- 2976981 TI - [Physical development of children in Astrakhan]. PMID- 2976982 TI - [Activities of regional agricultural medical teams]. PMID- 2976983 TI - [The main form of research conducted by students at medical institutes]. PMID- 2976984 TI - [The museum and its role in the training of physicians]. PMID- 2976985 TI - [A page from the history of social medicine]. PMID- 2976986 TI - [Improving the structure of regional sanitary-epidemiologic stations]. PMID- 2976987 TI - [Cancer of the thyroid gland in a goiter-endemic region]. PMID- 2976988 TI - [Analysis of mortality in acute peritonitis]. PMID- 2976990 TI - [Characteristics of the dynamic outpatient observation of workers at an industrial enterprise]. AB - A computer-assisted staged system of a follow-up of factory workers with CHD and diabetes mellitus was developed. It included automated screening by questionnaire, ECG and biochemical screening, physical examination, investigation of the basic carbohydrate-lipid indices, more detailed examination in out- or inpatients settings using up-to-date diagnostic methods (bicycle ergometry, monitoring after Holter, etc.), identification of groups for a follow-up, and therapeutic and sanitary measures. A total of 27750 persons were investigated. The most important group was that including persons with risk factors: vegetovascular dystonia developing into CHD (6%) or essential hypertension (20%). Sanitary measures by unified methods caused a decrease in temporary disability with the economic effect of 197,000 rubles. PMID- 2976989 TI - Mini-laparoscopy in blunt abdominal trauma. AB - Blunt abdominal trauma in multiorgan injured or comatose patients always presents a problem. The aim is to assess, in the shortest period of time, which organ injury requires priority and whether intra-abdominal bleeding or perforation exists. Abdominal lavage proved to be too sensitive. Not every positive case needs exploration. Approximately 15%-20% of the cases explored because of positive lavage did not show a significant bleeding site that would require surgical treatment. The authors developed a mini-laparoscope that can be used at the bedside, in the emergency room, or in the intensive care unit. The procedure can be performed with intravenous sedation and local anesthesia. In 150 cases, no hemoperitoneum was found in 53% of these cases. Except for 1, none of these patients needed further exploration. In 21%, severe hemoperitoneum was discovered; these patients were transferred to the operating room, and this was confirmed by surgery. In 26%, a small amount of blood was found in the gutters. These patients were observed in the intensive care unit and an unnecessary exploration was avoided. Laparoscopy gives a wider range of decision making by observing the abdominal cavity. It can be completed in 10-20 min at the bedside. No serious complications were encountered. This procedure should be taught and practiced in trauma centers. PMID- 2976991 TI - [The immune status in variants of the course and outcome of acute delta virus infection]. AB - The immune status was investigated in 32 patients with acute hepatitis of mixed etiology and in 39 HBsAg carriers, superinfected with delta virus. The relationship of changes in the immune status with a course and outcomes of acute delta virus infection was revealed. These changes returned to normal on patients with a cyclic course of disease, persisted in patients with a lingering process, and progressed in the development of subacute liver dystrophy. The most noticeable shifts (a decrease in all indices of the immune system) were revealed in patients with a fulminant course of disease. PMID- 2976992 TI - [Tuberculosis as the cause of death at the present stage of tuberculosis control]. AB - A 2-year study of all lethal outcomes in active tuberculosis in the Russian Federation has shown that 64.2% of patients died from progression of the specific process or its complications, 35.8%--from other diseases. A distinctive feature was that 42.8% of these were aged 20 to 49. The number of men exceeded that of women 5.8-fold. The negative factors were as follows: primary detection on consultation (47.9%), late diagnosis of far advanced forms of tuberculosis (23.4%), violation of the principles of chemotherapy by undisciplined patients, alcohol abuse and alcoholism among men (63.5%) and women (31.7%), concomitant diseases (58%). PMID- 2976993 TI - Modulation of heparin cofactor II function by S protein (vitronectin) and formation of a ternary S protein-thrombin-heparin cofactor II complex. AB - The complement inhibitor S protein, which is identical to the adhesive protein vitronectin, functions as heparin-neutralizing factor by protecting thrombin as well as factor Xa against fast inactivation by antithrombin III. The interference of S protein with glycosaminoglycan-catalyzed inhibition of thrombin by heparin cofactor II was investigated in these studies. S protein significantly counteracted the anticoagulant activity of heparin and pentosan polysulfate but not of dermatan sulfate. In the presence of 0.3 micrograms/ml heparin, 0.5 micrograms/ml pentosan polysulfate, or 2 micrograms/ml dermatan sulfate, S protein induced a concentration-dependent reduction of the inhibition rate of thrombin by heparin cofactor II. This resulted in a decrease of the apparent pseudo first-order rate constants by about 17-fold (heparin), or about 7-fold (pentosan polysulfate), whereas no neutralization of dermatan sulfate was demonstrable at a physiological ratio of S protein to heparin cofactor II. Exposure of the glycosaminoglycan-binding region of S protein by reduction and carboxymethylation of the protein increased the neutralizing activity of S protein towards heparin and pentosan polysulfate. The results of these functional experiments correlated well with the demonstration of direct binding of S protein to both polysaccharides but not to dermatan sulfate. While reduced/carboxymethylated S protein remained also ineffective in neutralizing other dermatan sulfate compounds with varying degree of sulfation, a synthetic highly basic tridecapeptide, representing a portion of the glycosaminoglycan binding domain of S protein, counteracted their anticoagulant activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2976994 TI - Fibrin metabolism in patients with acute myocardial infarction during and after treatment with tissue-type plasminogen activator. AB - In order to define some of the determinants of successful thrombolysis and reocclusion during fibrinolytic therapy for acute myocardial infarction (AMI), specific molecular markers of fibrin metabolism were serially measured in 15 patients with AMI treated with tissue-type plasminogen activator (t-PA). Fibrin formation was assessed by measurement of fibrinopeptide A (FpA) and fibrinolysis by assay of B-beta peptides 1-42 and 15-42 and crosslinked fibrin degradation products (XDP). At baseline, FpA levels were high while markers of fibrinolysis were near normal. Following a 90-minute infusion of t-PA (0.5-1.1 mg kg-1 hr-1), all markers of fibrinolysis increased. Levels of FpA remained elevated despite heparin at the initiation of cardiac catheterization. None of these markers discriminated between patients with successful reperfusion from those without. At 4 hours, B-beta 15-42 peptide and XDP levels remained elevated suggesting persistence of fibrinolysis beyond the short circulatory half-life of t-PA. FpA levels at 4 hours were lower in patients who underwent acute coronary angioplasty compared to those who received additional low dose t-PA (12.3 +/- 4.5 vs. 30.4 +/ 5.5 ng/ml, p less than 0.05). By 48 hours, markers of fibrinolysis had returned toward normal except in 2 patients with persistently elevated B-beta 15-42 peptide levels who suffered reocclusion on days 5 and 6 (75 and 44 vs. 29 +/- 3 nM, p less than 0.005). In conclusion, molecular markers of fibrin metabolism during fibrinolytic therapy may provide clinically relevant data. PMID- 2976995 TI - Normal plasmic cleavage of the gamma-chain variant of "fibrinogen Saga" with an Arg-275 to His substitution. AB - We have identified a gamma-Arg-275 to His substitution in an abnormal fibrinogen designated as "fibrinogen Saga" characterized by impaired fibrin monomer polymerization. By chromatofocusing chromatography, we isolated normal and abnormal fragment D1 populations separately from the plasmic-calcium digests of fibrinogen derived from the propositus, a heterozygote for the abnormality. We found that both normal and abnormal fragment D1's were similarly protected from digestion by plasmin in the presence of calcium ions and further degraded to fragments D2 and D3 due to cleavage of the gamma-chain remnant when calcium ions were replaced by chelating agents. Abnormal fragment D1 failed to inhibit both thrombin-clotting of normal fibrinogen and polymerization of normal fibrin monomer, while normal D1 exhibited marked inhibitory activities. In an aberrant peptide comprising residues gamma-274-302 isolated by HPLC from the lysyl endopeptidase-digests of abnormal fragment D1, we identified a His substituting for an Arg at position 2, which corresponds to position 275 of the mutant gamma chain. PMID- 2976996 TI - [Backache in children and adolescents]. AB - Although backpain during childhood is uncommon serious diseases feature high on the list of differential diseases. Often the correct diagnosis in the young is made late or missed in the first instance. During the adolescent period the incidence of backpain rises to include mechanical or osteochondritic conditions and the early problems of a degenerative back such as disc protrusion. It has to be stressed that infection and neoplasma although rare can be a cause of backache at any age. If after thorough investigation no abnormalities has been detected, one should keep in mind that serious mental problems can be the cause of backpain. PMID- 2976997 TI - The production of tumor necrosis factor-alpha by rat basophilic leukemia cells with triggering IgE receptor. AB - We evaluated cytotoxic factor released from rat basophilic leukemia cells (RBL) sensitized with anti-ovalbumin (OVA) mouse serum after incubation with OVA. The cytotoxic activity of this factor was completely blocked by anti-mouse tumor necrosis factor-alpha (mTNF-alpha) specific antibody. Therefore, we concluded that by triggering the IgE receptor, RBL could produce and release TNF-alpha. PMID- 2976998 TI - [Indices of the local volumetric blood flow and oxygenation in hemangiomas of the face and neck]. PMID- 2976999 TI - [The incidence of partial tooth loss among the population of the Moldavian SSR]. PMID- 2977000 TI - [Ideal indications for the use of a textile ligament of high strength Trevira in alloplastic replacement of the anterior cruciate ligament in chronic isolated rupture]. AB - Between August 1980 and December 1986, 120 patients suffering from chronic instabilities of the knee ligament were submitted to corrective surgery at the Berufsgenossenschaftliche Unfallklinik Frankfurt am Main. A textile ligament made of polyethylene terephthalate (Trevira hochfest type 730) was substituted in all cases for the disturbed or missing ligament structure. 67 out of 100 patients came to the check-up appointments. A precise and objective comparison between the preoperative and postoperative findings of each patient was now possible for the first time thanks to the most recent computer technique. Until now we were not able to do this. Possibly the results published hitherto had a subjective touch and were too much dependent on the feelings of the individual investigators. Full objective stability over several years was reached anyhow in 51.4% of the patients checked up. Especially in case of inveterate isolated rupture of the anterior cruciate ligament, the alloplastic substitution by a ligament made of Trevira hochfest is at present an ideal possibility to restore stability for a long time and to prevent severe secondary wear symptoms of the other interior structures of the knee. The failures and insatisfactory results with respect to stability are explained by an insufficient consideration of the implantation technique recommended. In order to help this method to achieve its welldeserved propagation which would be to the advantage of patients with chronic instabilities of the knee ligament, this technique is explained in detail once more and possible errors are indicated. PMID- 2977002 TI - Passive lumbar mobility. A prospective study of back pain in young men during their military service. AB - Mobility of the back is mostly evaluated through active mobility in a standing position. In this study, also the passive mobility between lumbar vertebraes was assessed in sidelying, and grouped in four categories. More than five hundred young men were examined three times over a period of 3-4 years, before and after their basic military training. The lumbar vertebra L5 was judged to have decreased mobility in 29% and increased mobility in 17% of the cases. The corresponding values for L4 were 39% and 13%, respectively. There were significant positive correlations between the results of all the examinations. The total agreement between examinations was around 50%. Decreased passive lumbar mobility correlated to the current amount of back discomfort at the second and third examinations, but at the first examination it could not predict the future incidence of back pain. PMID- 2977001 TI - Leg length inequality. A prospective study of young men during their military service. AB - Within a prospective study of back function and pain before and after basic military training, the leg length inequality (LLI) was assessed, in steps of less than 0.5 cm. (equal), 0.5-1.5 cm, 1.6-2.5 cm, 2.6-3.5 cm and more than 3.5 cm. Around six hundred young men were examined three times over a period of four years. LLI of 0.5-1.5 cm was found in 32%, and 4% had a difference of over 1.5 cm. Pelvic rotation was noted in 15% of the cases. The average total agreement of identifying LLI was 64% between the three examinations. No correlation was found between LLI and back-pain or pain-provocing tests. In those with LLI in standing there was a tendency towards more remarks on SI-joint mobility tested in lying. During the follow-up period, no correlation of the LLI and the result of the other examination variables could be found. PMID- 2977003 TI - Tightness of hamstring- and psoas major muscles. A prospective study of back pain in young men during their military service. AB - Muscular tightness and the therapeutic effect of stretching has been widely discussed during the last few years in sports training and physiotherapy. Within a prospective study of back function and pain before and after compulsory military service, tightness of hamstring- and psoas muscles was assessed. Around 600 young men were examined three times over a period of four years. Tight hamstring muscles were found to be very common in this group. Only 43% of the right and 35% of the left legs reached an angle of at least 80 degrees from the couch during the straight-leg-raising test (Lasegue's test). The test of muscular tightness showed a significant test-retest reliability over all examinations. Tight hamstring- or psoas muscles could not be shown to correlate to current back pain or to the incidence of back pain during the follow-up period. PMID- 2977005 TI - [Early and late results of dilatation of the coronary vessels]. PMID- 2977004 TI - Zinc ion stimulation of ATP cleavage by prostasomes from human seminal plasma. AB - A stimulation by zinc ions of the hydrolysis of ATP by prostasomes prepared from human semen has been observed. Stimulation was maximal at a Zn2+/ATP stoichiometry of 0.5/l, and increasing this ratio resulted in a gradual decrease in ATPase activity. The pH optimum was 6.0. The apparent Km for Zn2+-dependent ATPase was 0.43 mmol/l and apparent Vmax 5.60 mumol/mg protein/20 min. Other divalent cations could replace Zn2+ as cofactor more or less effectively in the order Mn2+ greater than Cd2+ greater than Ba2+ greater than Sr2+. Potassium ions produced a further activation of the Zn2+-dependent ATPase system by about 10%. Such a stimulation was also attained to some extent by other monovalent cations as Rb+, NH4+, Li+ and to a lesser extent by Cs+. Orthovanadate in the concentration interval 5-1,000 mumol/l was inhibitory of the Zn2+-dependent ATPase system in a dose-dependent fashion. An aminopeptidase activity was also linked to the prostasomes. This enzyme activity was dramatically inhibited by 2 mmol/l orthophenantroline. A reactivation of the orthophenantroline-inhibited aminopeptidase activity was possible by adding Zn2+ to the reaction mixture. Hence, prostasomes contained ATPase as well as aminopeptidase activities both of which being dependent upon Zn2+. These two activities did not seem to be expressions of an ATP-dependent protease activity associated with prostasomes. PMID- 2977006 TI - [Coronary risk factors at the work site. Identification, prevention, compensation]. PMID- 2977007 TI - [Relaparotomy: its meaning and classification]. PMID- 2977008 TI - [A "semi-open" method of treating diffuse suppurative peritonitis]. PMID- 2977009 TI - [Various causes of high rate of mortality in incarcerated hernia]. PMID- 2977011 TI - [Surgical treatment of arteriosclerosis at the Surgical Disease Clinic No. 2 (head: Prof. L.V. Lebedev) of the Pavlov First Medical Institute in Leningrad]. PMID- 2977010 TI - [Causes of insufficient reduction of mortality rate after operations for incarcerated external ventral hernia]. PMID- 2977012 TI - [Nikolai Nikolaevich Samarin (on the centenary of his birth)]. PMID- 2977013 TI - [Clinico-roentgenologic characteristics of calcinosis and ossification of postoperative scars of the white line of the abdominal wall]. PMID- 2977014 TI - [Structural and functional changes in myosin from rat skeletal muscles during postmortem autolysis]. AB - Structure and enzymatic properties of myosin were studied in rat skeletal muscles during various periods of postmortal autolysis. Ca2+-ATPase activity of the protein was decreased within 3 hrs after death. At the same time, the structure of myosin, as shown by gel filtration in 8 M urea and disc electrophoresis in SDS polyacrylamide gel, was maintained. The phenomenon of "substrate inhibition" exhibited both native myosin and the protein isolated from the autolyzed muscle tissue. Enzymatic activity of myosin was markedly decreased within 12 hrs and 24 hrs after death. Degradation of the protein heavy chains was also observed. PMID- 2977015 TI - [Experience in organizing nephrologic and urologic care for the population of Orel Province]. PMID- 2977016 TI - [Factors influencing the development of myocardial hypertrophy]. PMID- 2977017 TI - [Echocardiographic evaluation of cardiac function in the diastole of hypertension patients with various degrees of left ventricular hypertrophy]. PMID- 2977018 TI - [Pathophysiology of polysaccharide sulfates in the connective tissue]. PMID- 2977019 TI - [Multiple trauma with injury of the heart, lung, liver, abdominal wall, face and right arm]. PMID- 2977020 TI - [Early reaction of rejection of esperal after implantation]. PMID- 2977021 TI - [Various elements of the experimental introduction of the Book of Health for retired and handicapped persons in an urban environment]. PMID- 2977022 TI - [Endometriosis of the abdominal wall scar after cesarean section]. PMID- 2977023 TI - [Ultrasonographic characteristics of asymmetrical hypertrophy of the interventricular septum in patients with arterial hypertension]. PMID- 2977024 TI - [Coexistence of congenital heart defects with other congenital abnormalities]. PMID- 2977025 TI - [Use of free submerged autologous skin transplants for the repair of abdominal hernia. Early and remote results]. PMID- 2977026 TI - [New aspects of nuclear medicine diagnosis of kidney function: improved potential by pharmacologic intervention and quantitative analytic procedures]. AB - In nuclear medicine new trends in the diagnosis of renal function are based on the introduction of new radiopharmaceuticals, improvements in the methodological part of the procedure and precise pharmacological intervention in response to given indications. Tc99m mercaptoacetyltriglycine (Tc99m MAG3) was tested as replacement for I123 orthoiodohippuric acid (I123 oIH) both in the form of a HPLC purified substance and as an impure kit preparation. HPLC purified Tc99m MAG3 clearance determinations in anuric patients showed a low extrarenal excretion amounting to only about 5% of the total clearance in normal patients. Kit preparations yielded about 90% of the labelled product; impurities were pertechnetate, reduced hydrolyzed Tc99m and chemically unidentified labelled products which showed a significantly lower renal, but increased hepatobiliary excretion in comparison with Tc99m MAG3. The renal clearance with kit preparations of Tc99m MAG3 was 55% of the clearance with oIH at a comparable urinary excretion. Significantly higher protein binding and therefore, a decrease in the distribution volume of Tc99m was found in comparison with I123 oIH. No difference was recorded between the two substances with respect to the renogram curves in normal subjects, apart from a modest delay in the elimination of Tc99m MAG3. For clinical purposes kit preparations of Tc99m MAG3 proved equal to I123 oIH. The influence of angiotensin converting enzyme (ACE) inhibitors (captopril) leads to characteristic changes in the renograms of patients with Goldblatt hypertension. Quantitative criteria for the evidence of haemodynamically significant renal artery stenosis were derived from investigations without and with captopril (25 mg) (I123 oIH and Tc99m DTPA) in 21 patients with essential hypertension. The criteria were defined as follows: a delay in peak activity (Tmax) in the I123 oIH captopril renogram exceeding 2 minutes as compared with the baseline value and/or a lower uptake of Tc99m DTPA in comparison with the uptake of I123 oIH (uptake quotient I123 oIH/Tc99m DTPA greater than 1.2). The diagnostic and prognostic potential of the captopril renogram was compared with that of the captopril test by investigating 34 patients with renal artery stenosis (23 uni-, 11 bilateral) (atherosclerosis: 23, fibromuscular hyperplasia: 11). The captopril renogram was positive more often (n = 12) than the captopril test (n = 4) in patients without renal functional impairment of the stenosed kidney. Similar results were obtained with both methods in patients with atrophic kidneys: captopril renography was positive in all cases with a positive captopril test.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2977027 TI - [Effectiveness of aerosol treatment with spasmolytic agents of obstructive bronchitis in infancy and early childhood]. AB - In 64 children in the age of 3 months to 3 years with obstructive bronchitis the effectivity of aerosol treatment with orziprenalin and the combination of fenoterol with ipratropium bromide (Berodual) using ultrasonic nebulizer, the treatment with Berodual metered pressure aerosol, and the effectivity of the inhalation of physiological (isotonic) saline solution during acute respiratory virus infection has been compared. The effectivity was evaluated by clinical observation and on the base of the determination of capnographic and oxymetric values. Compared to the controls (inhalation of saline), a very high effectivity of all 3 bronchospasmolytic formulations has been observed. The ease of the use of the Berodual metered pressure aerosol is stressed. PMID- 2977028 TI - [Etiology of pathogen-induced changes between the toes in patients working under moist conditions]. AB - Referring to recent studies, we discuss the epidemiology of tinea pedis and its position within the range of polyetiological foot infections caused by microbial agents. 201 industrial workers used to wear rubber boots because of moist working conditions were examined with regard to their feet. 107 of them showed clinically altered toe web; in 45 cases, we found ringworm infection (22.4%), in correlation to increased daily wearing time of rubber boots. 85 workers showed colonization of pathogenic bacteria (42.4%), especially gram-negative species (25.8%). Frequently, mixed infections were found. The clinical picture not always allows definite conclusions as to the causal agents. The symptom "maceration" is seen in each of the 3 pathogenic groups of germs (fungi, gram-positive pathogenic cocci, gram-negative bacteria), frequently in bacterial foot infections, especially gram negative infections. Marked clinical symptoms involve increased infection rates. The predisposing factors to foot infection such as interdigital maceration and the promoting role of rubber boots have been confirmed. The final diagnosis is to be reassured by cultural investigations, in order to set up a specific treatment. PMID- 2977029 TI - [Hemodynamic, anti-ischemic, metabolic and neurohumoral effects of enoximone (MDL 17,043) in patients with coronary disease]. AB - The hemodynamic, anti-ischemic, metabolic, and neurohumoral effects of the phosphodiesterase inhibitor enoximone were investigated in 17 patients (mean age 58 +/- 2 years) with coronary heart disease as established by coronary angiography and positive exercise tests after i.v. application of 0.75 mg/kg body weight. Whereas administration of enoximone resulted in a significant increase in heart rate from 75 +/- 17 to 83 +/- 14 per minute (p less than 0.01), exercise heart rate, blood pressure and myocardial oxygen consumption did not change significantly (p greater than 0.05). At rest, enoximone led to a significant decrease of mean right atrial pressure from 5.7 +/- 2.3 to 3.8 +/- 1.2 mm Hg (p less than 0.01). During exercise there was a significant fall in pulmonary pressure (PAm from 40 +/- 7 to 24 +/- 7 mm Hg, p less than 0.001; PCm from 24 +/- 7 to 14 +/- 6 mm Hg, p less than 0.001) caused by preload reduction and concomitant inotropic increase; there was also a significant rise in cardiac output from 12.7 +/- 5 to 13.8 +/- 5 mm Hg (p less than 0.01) and a decrease of ST-segment depression from 1.97 +/- 0.76 to 0.53 +/- 0.51 mm (p less than 0.001). With improved peripheral and probably coronary blood flow, a concomitant decrease of the metabolic ischemic markers was detected during exercise (potassium 4.44 +/ 0.29 vs. 4.31 +/- 0.30 mval, p less than 0.05; lactate 19 +/- 9 vs. 18 +/- 7 mg/dl; pH 7.28 +/- 0.27 vs. 7.36 +/- 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977030 TI - Combined evaluation of postphlebitic limbs by laser doppler flowmetry and transcutaneous PO2/PCO2 measurements. PMID- 2977032 TI - [The role of heredity in the development of chronic viral hepatitis]. PMID- 2977031 TI - [Does the action of regulatory peptides introduced exogenously correspond to their physiologic functions in the plasma and cerebrospinal fluid?]. PMID- 2977033 TI - [The opiate system of the brain in patients with alopecia areata]. PMID- 2977034 TI - [Nosology, epidemiology and prevention of mycosis caused by T. rubrum]. PMID- 2977035 TI - [Venereal disease morbidity among pupils]. PMID- 2977036 TI - Prenatally diagnosed gastroschisis--a preliminary report advocating the use of elective caesarean section. AB - Seven cases of prenatally diagnosed gastroschisis were evaluated. All infants were delivered by elective Caesarean section and brought to the operating theatre immediately after birth where the abdominal wall defect was closed within one hour post partum. Primary closure was easy and successful in all cases without widening of the defect and with the umbilicus left intact, because of the total lack of oedema and bowel peel. The average duration of postoperative total parenteral nutrition (7 days) and hospital stay (18 days) was significantly shorter in this prenatally diagnosed group compared to traditionally treated cases. PMID- 2977037 TI - [Fatal lung hemorrhage in complex pulmonary angiodysplasia in a case of mongolism]. AB - A case of sudden and unexpected death of a boy with Down-Syndrome is reported. Death was due to acute excessive haemorrhage from the lungs. There were no cardiac anomalies. Histology revealed in addition to mediahyperplasia, intimaproliferation and occlusions, multiple foci of anomalous ectatic blood vessels predominantly in the lungs, some of which ruptured and bled into adjacent airways. Evidence of previous less severe bleeding episodes was present. Further investigation showed a familial occurrence of the blood vessel anomaly. PMID- 2977038 TI - Peritonitis and massive granulocytic infiltration of the spleen in adult Still's disease. AB - A case of adult Still's disease is described which, in addition to the more common manifestations, also included abdominal discomfort. Upon laparoscopy, peritonitis was disclosed; a biopsy showed massive granulocytic infiltration of the spleen which could not be attributed to an infectious disease. The patient did not improve on conventional therapeutic modalities but required intensive combination therapy consisting of high dose acetylsalicylic acid, prednisone, and slow-reacting substances before entering remission. PMID- 2977039 TI - [HLA-B27-negative sacroiliitis as a complication of inflammatory forms of acne: case reports and review of the literature]. AB - In this review, three patients with severe inflammatory acne accompanied by a distinct rheumatologic syndrome are discussed together with 44 similar cases reported in the literature. This syndrome, here referred to as "acne-associated arthritis (AAA)" usually involves male teenagers and twins. The most characteristic clinical findings relate to the axial skeleton as well as the sterno-costo-clavicular region. Although no association with HLA-B27 has been found, AAA shares a number of clinical, radiological and histomorphological features with seronegative spondylarthropathies. Furthermore, nosological connections seem to exist with a number of ill-defined "hyperostotic osteoarthropathies." Similar characteristics are known from other "pustulotic or neutrophilic dermatosis." The hypothesis of an infectious etiology of AAA has not been substantiated. Responses to systemic antibiotics are generally disappointing, whereas steroids have been shown to exhibit some therapeutic efficacy. PMID- 2977040 TI - [Composite adhesive technique. Precise-sure-patient justified]. PMID- 2977041 TI - [Schon's acrylic bite. A simple method for bite registration]. PMID- 2977042 TI - [Are nickel based alloys dangerous?]. PMID- 2977043 TI - [The medroxyprogesterone acetate serum level following various medroxyprogesterone acetate dose schedules in gynecologic oncology]. AB - Medroxyprogesterone acetate (MPA) is used as an adjuvant hormonal medication in patients with different kinds of carcinomas. Since adequate serum levels are thought to be essential we determined the individual, postoperative MPA levels in 36 patients with endometrial carcinoma over a period of 12 weeks. The patients received either an oral dose of 3 X 100 mg MPA or a weekly changing scheme with 2 X 10 mg Tamoxifen and 3 X 100 mg MPA. An additional small group of 4 patients with ovarian carcinoma was enrolled receiving an oral dose of 1000 mg MPA daily. The peripheral serum levels of MPA exhibit enormous inter- and intraindividuell variations and only the high dosage schemes yield levels above 90 ng/ml which are claimed necessary by some authors. The cortisol concentration measured at the same time were within the normal range and did not correlate with the MPA values. PMID- 2977044 TI - [The training of neurosurgeons in clinical internships]. PMID- 2977045 TI - [Interprovince neurosurgical centers in the RSFSR]. PMID- 2977046 TI - [Indications for lumbar facet infiltrations. Preliminary results]. PMID- 2977048 TI - [Cerebral palsy: its effects on eruption of the first permanent molar and health illness condition]. PMID- 2977049 TI - [The Zambrano Gomez technic for reduction of the size of the tongue]. PMID- 2977047 TI - Laminar distribution of neuritic plaques in normal aging, Alzheimer's disease and Down's syndrome. AB - Quantitative studies of neuritic (senile) plaques in six cortical layers were carried out in brains from people with confirmed clinical and neuropathological diagnosis of senile dementia of the Alzheimer type (SDAT) and Down's syndrome (DS). The same studies were performed on brains of normal old-aged people. In Alzheimer disease (AD) and DS cases the highest numbers of neuritic plaques (NP) were observed in temporal lobe layers III and II and occipital lobe layers III, IV and II. In normal old-aged people the highest numbers of NP were found in temporal lobe III and V and in occipital lobe IV, III, and V layer. The plaque numbers in both temporal and occipital cortices of AD and DS were significantly higher than that of normal old-aged people but there was no difference between AD and DS. PMID- 2977051 TI - Laboratory tests as indicators of alcohol consumption in patients with chronic low back pain. AB - The use of laboratory screening tests for the detection of heavy drinking in outpatients implies among other things knowledge about their relation to the degree of alcohol consumption. In this study of 95 patients with chronic low back pain, serum gamma-glutamyltransferase demonstrated the best correlation with the mean daily amount of alcohol consumed (r = 0.74). Adding other serological tests, such as high density lipoprotein cholesterol, aminotransferases and linoleic acid content in lecithin, did not improve this correlation, although these tests also correlated well with the alcohol intake. PMID- 2977050 TI - Caffeine stimulates in vivo platelet reactivity. AB - The effect of coffee drinking on platelet reactivity was studied in 12 healthy subjects. Plasma beta-thromboglobulin concentration was determined before and one hour after administration of 100 mg of caffeine, corresponding to one cup of coffee. Mean values were 47.0 +/- 19.3 and 179.3 +/- 85.5 ng/ml before and after caffeine administration respectively. The increase, 298 +/- 150%, is highly significant (p less than 0.001). PMID- 2977052 TI - Incidence and outcome of reintervention after coronary bypass surgery. PMID- 2977053 TI - Thrombolysis in acute myocardial infarction and the indication for aortocoronary bypass surgery. PMID- 2977054 TI - Immediate bypass surgery (0.5-3 h) for evolving myocardial infarction. PMID- 2977055 TI - Emergent surgical revascularization following acute myocardial infarction. Experimental and clinical considerations. PMID- 2977056 TI - Removal of extrauterine IUDs by the vaginal route with laparoscopic assistance. AB - Although laparoscopy is the preferred method of removing extrauterine IUDs, occasionally removal is complicated, especially if the IUD is encased in omentum. This paper reports on five such cases in which the extrauterine IUDs were removed vaginally through a posterior colpotomy with laparoscopic assistance. PMID- 2977057 TI - [Outpatient care of handicapped children and adolescents]. PMID- 2977058 TI - [Some aspects of pediatric and specialist outpatient management of preschool and school children (3-18 years of age)]. PMID- 2977059 TI - [Biochemical and immunological properties of 34kDa EDTA-extractable protein (EEP34k)]. PMID- 2977060 TI - [Clinical evaluation of hochu-ekki-to on the patients with renal ptosis]. AB - Hochu-ekki-to was administered orally in 2.5 g doses thrice per day to 53 patients who complained of lumbago or lower abdominal discomfort. Its efficacy rate was 53.8% for the patients who complained of lumbago, and 32.3% for the patients who complained of lower abdominal discomfort. Total efficacy rate was 66.0%. Mild adverse effects were observed in 4 patients, but no severe untoward effects was observed. PMID- 2977062 TI - [Anesthetic management in dentistry for severely handicapped patients in Okinawa ken]. PMID- 2977063 TI - [Tactics in the management of pregnancy complicated by lumbar vertebrogenic pains]. PMID- 2977061 TI - Serial cell-mediated immunological changes in terminal uremic patients on continuous ambulatory peritoneal dialysis therapy. AB - To investigate the mechanism of immunologic defects in uremia, we examined the cell-mediated immunity in 20 uremic patients with well balanced dietary assessments before continuous ambulatory peritoneal dialysis (CAPD) therapy and the serial changes in cell-mediated immunity 3, 6, 9, and 12 months after therapy. Absolute lymphocyte count, active T, total T, OKT4, OKT8, and B-cell levels were significantly lower in uremic patients than in controls, but progressively increased after CAPD treatment. The lymphoproliferative responses to mitogens decreased and were more evidently suppressed in the presence of autologous plasma. The suppressive effect of autologous plasma could be abolished by indomethacin treatment. The plasma prostaglandin E2 level, which was increased in uremic patients, decreased after CAPD treatment. These results suggest that prostaglandin E2 may play an important role in the suppression of T-cell function. The T-cell response to the stimulation of autologous non-T cells (autologous mixed-lymphocyte reaction; AMLR) in uremic patients was also low in patients before CAPD treatment. The patients' autoreactive T cells could suppress AMLR, and so did the supernatant from AMLR and isolated T-cell culture of uremic patients. These results suggest the existence of suppressor autoreactive T cells which can release suppressor factor(s). However, to our surprise, the autoreactive T-cell proliferation increased, and the suppressive effect of the supernatant was abolished after CAPD therapy. The mechanism for this peculiar phenomenon is not known and deserves further investigation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977064 TI - [The main trends in the activities to improve mass screening of women]. PMID- 2977065 TI - [Predicting professional competence of subinterns based on an analysis of personality characteristics]. PMID- 2977066 TI - Determination of glutathione in biological material by flow-injection analysis using an enzymatic recycling reaction. AB - A sensitive and specific assay for glutathione using a recycling reaction followed by spectrophotometric detection in a flow-injection analysis system is presented. The proposed method provides specific amplification of the response to glutathione by combined use of the enzyme GSSG reductase and the chromogenic reagent 5,5'-dithiobis(2-nitrobenzoic acid). Both oxidized (GSSG) and reduced (GSH) glutathione are detected, so that GSSG must be determined separately after alkylation of the GSH with N-ethylmaleimide. The sensitivity is controlled by the number of times the cycle occurs and therefore by the residence time of the sample in the reactor. This time depends on the reactor length and the flow rate. The influence of residence time, temperature, and enzyme concentration on the response has been studied and the optimum reaction conditions have been selected. The sample throughput is as high as 30 h(-1) and the detection limit is 1 pmol GSH at a signal-to-noise ratio of 3. The method has been evaluated by the quantification of GSH and GSSG in isolated hepatocytes. A high correlation between the new flow-injection analysis method and the original spectrophotometric batch assay has been found (slope = 1.039, intercept = 0.6, n = 216, r = 0.977). The main advantages of the proposed method are high sample throughout, high sensitivity, and good reproducibility. PMID- 2977067 TI - Dot-blot assays and their use as a direct antigen-binding method to screen monoclonal antibodies to 1,4-beta- and 1,3-beta-glucan synthases. AB - A rapid method has been developed to assay beta-glucan synthases spotted on a nitrocellulose sheet. The sensitivity of this method allows screening of hybridoma-making monoclonal antibodies in a direct antigen-binding assay by measurement of the activity of the enzymes retained by the antibodies previously fixed on nitrocellulose. PMID- 2977069 TI - Enhancement of uphill transport by a double carrier membrane system. PMID- 2977068 TI - Plaque-lift testing of expression vector lambda gt11 with gold-labeled immunoglobulins. AB - Colloidal gold particles were coated with affinity-purified antibodies against the human plasma protein, C1 inhibitor, and used to probe for fusion proteins of C1 inhibitor with beta-galactosidase encoded by recombinant bacteriophage lambda gt11 DNA. Plaque-lift tests were done with recombinant proteins immobilized on nitrocellulose applying anti-C1 inhibitor gold particles followed by the silver enhancement treatment. This procedure resulted in a sensitive and specific staining of the recombinant proteins and allowed the selective detection of relevant clones in a complex cDNA expression library. Under optimized conditions, plaque-lift testing was completed within 2.5 h after removal of nitrocellulose filters from the plate. Hence, the immunogold detection method provides an alternative to conventional enzyme- or radionuclide-based screening procedures for cDNA expression libraries. PMID- 2977070 TI - Efficacy of febantel against abomasal nematodes and lungworms in cattle. AB - The efficacy of febantel at a dosage of 5 mg/kg (45.5% paste formulation) against inhibited early 4th-stage larvae (EL4) of Ostertagia ostertagi, other nematodes of the abomasum, and Dictyocaulus viviparus was investigated in 4- to 6-month-old Holstein calves that grazed on pasture heavily contaminated with parasites from February 24 to April 1, 1986 (36 days). In Louisiana, this is the first month of a 3-month period in which increasing numbers of inhibition-prone O ostertagi larvae are acquired, and infection risk with D viviparus may remain high. Three of 4 calves that died of lungworm infection during the pasture-exposure period were necropsied. Large numbers of abomasal nematodes, including inhibited O ostertagi larvae, and large numbers of D viviparus were recovered. Twenty-five calves were randomly allotted by equal distribution of body weight to 2 groups and treated on April 4: placebo-treated calves (n = 13) and febantel-treated calves (n = 12). Equal numbers of treated and control calves were killed at 6 and 7 days, respectively, after treatment. Mean numbers of O ostertagi in control cattle were: adults, 4,931; developing 4th-stage larvae (DL4), 1,119; and inhibited EL4, 3,410. Ostertagia lyrata, Trichostrongylus axei, Haemonchus sp, and D viviparus were well distributed in nearly all control calves. Percentage reduction of O ostertagi in treated calves, when compared with controls, was: adults, 83.6%; DL4, 57.8%; and inhibited EL4, 34.8%. Percentage reductions of other species were: O lyrata, 92.6%; T axei adults, 99.3% and 4th-stage larvae (L4), 100%; Haemonchus sp adults, 66.7%, and L4, 64%; D viviparus adults 90.6%, and immature forms, 97.1%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977071 TI - [Erythrokeratoderma variabilis (Mendes Da Costa). Ultrastructural study]. PMID- 2977072 TI - [Mendes Da Costa erythrokeratoderma variabilis. Effect of RO 10-9359 (Tigason)]. PMID- 2977073 TI - [Harlequin fetus treated with etretin (RO 10-1670)]. PMID- 2977074 TI - [Filiform palmar hyperkeratosis and rectosigmoid cancer]. PMID- 2977075 TI - [Cutaneomucous adverse effects of continuous infusion of 5-fluorouracil: 12 cases]. PMID- 2977076 TI - [Severe drug accidents and allopurinol]. PMID- 2977077 TI - [Lichenoid dermatitis with alopecia caused by quinidine]. PMID- 2977078 TI - [Deferoxamine (Desferal) dermatitis]. PMID- 2977079 TI - [New iatrogenic acne: acne caused by amineptin (Survector)]. PMID- 2977080 TI - [Iatrogenic acne caused by amineptin (Survector). Apropos of 8 cases]. PMID- 2977082 TI - [Acne caused by Survector]. PMID- 2977083 TI - [Monstrous acne in the adult. Inducer role of Survector?]. PMID- 2977084 TI - [Rosaceous drug eruption caused by amineptin (Survector)]. PMID- 2977081 TI - [Severe lesions of acne type induced by chronic amineptin poisoning: apropos of 6 cases]. PMID- 2977085 TI - [Giant HPV 6 Buschke-Lowenstein tumor with metastases in the abdominal wall]. PMID- 2977086 TI - Cerebrospinal fluid levels of quinolinic acid in Huntington's disease and schizophrenia. AB - The concentration of the endogenous excitotoxin quinolinic acid was determined in the cerebrospinal fluid of drug-free patients suffering from Huntington's disease or schizophrenia (control group). In both diseases, quinolinic acid concentrations were highly variable (less than 4-48 nM) but the mean levels for each disease group were not significantly different from each other or from the quinolinic acid concentration of normal cerebrospinal fluid. Analysis of steady state cerebrospinal fluid quinolinic acid concentration is unlikely to be of value as a diagnostic tool in Huntington's disease. PMID- 2977087 TI - DNA double-chain breaks in recombination of phage lambda and of yeast. PMID- 2977089 TI - Compliance with risk factor reduction among post-coronary-bypass surgery and post coronary angioplasty patients. PMID- 2977088 TI - Spliceosomal snRNAs. PMID- 2977090 TI - [Immunology of transplantation]. AB - Immunologic events leading to the acute rejection of a grafted organ follow three sequential stages: 1) recognition of the antigens fully specific expressed by the allograft; 2) proliferation and differentiation of the T lymphocytes; 3) destruction of the graft mediated by various cellular types: sensitized cytotoxic T lymphocytes, lymphokine activated lymphocytes or macrophage cells, and, more hypothetically, by the so-called "killer" cells. The T lymphocytes infiltrating the grafts during rejection include functionally distinct subsets: inducer T cells (CD4+ cells) and suppressor/cytotoxic cells, T cells (CD8+ cells). In reversible acute rejections the role of activated cytotoxic cells (CD3+ CD8+) seems to be predominant. The functional nature of the lymphoblasts infiltrating the allograft is heterogeneous. It includes cell-mediated cytotoxicity, the proliferative response to alloantigens and the production of lymphokines. Finally, the role of the T cell receptor for the recognition of the antigen appears to be essential. The necessity for the immune system to build up a large repertoire of different T receptors is dependent on a mechanism of genetic coding similar to the one used by immunoglobulins. In the near future, one may hope to alter specifically the repertory of the T receptors used by the lymphocytes responsible for rejection. This would represent the dawn of a new era in transplantation immunity with the beginning or specific immunosuppression. PMID- 2977092 TI - Ultrasound directed oocyte recovery in an IVF program. PMID- 2977091 TI - [Thrombolysis with tissue plasminogen activator in patients with acute myocardial infarction. Preliminary report]. PMID- 2977093 TI - Acne in the adolescent. PMID- 2977094 TI - Depo-Provera--missing the point? PMID- 2977095 TI - Progestogens for contraception. PMID- 2977096 TI - Traumatic skin conditions in the athlete. PMID- 2977097 TI - Genetic control of susceptibility to severe hyperglycaemia evoked by CFA/SZ induced immune response against beta cells in various rat strains. AB - A novel approach has previously been reported to induce an insulin-dependent (type 1) diabetes mellitus in Wistar and Lewis rats by subdiabetogenic dose of the beta cell toxic agent streptozotocin injected i.p. 24 h after a polyclonal activator of lymphocytes, the complete Freund's adjuvant, was administered. The results from a comparative study in Wistar rats and congenic Lewis rats of the haplotype RT1a and RT1u demonstrate that both the genetic background and genes linked to the major histocompatibility complex are probably involved in the genetic control of susceptibility to the induction of hyperglycaemia in this experimental diabetes model. The susceptibility to hyperglycaemia was strongly associated to the non-specific activation of the immune system by administration of complete Freund's adjuvant only. From these data it is suggested that the genetic control of diabetes induction in this animal model is caused by genetic control of the immune/autoimmune reactivity involved in the mechanisms of beta cell destruction. PMID- 2977099 TI - The Neurospora am gene and allelic complementation. PMID- 2977098 TI - Studies on the fast reacting thiol groups in phosphofructokinase from baker's yeast. AB - The fast reacting thiol groups of yeast phosphofructokinase were studied by means of stopped-flow measurements. The enzyme was found to contain four very fast reacting cysteinyl residues determined by their reactivity towards 5,5' dithiobis(2-nitrobenzoic acid). A second class of eight thiol groups reveals an apparent first order rate constant which is three orders of magnitude lower than the rate constant of the first one. Due to the extreme high reactivity of the first class of cysteinyl residues partial oxidation was already observed under aerobic conditions. Fructose 6-phosphate, fructose 1.6-bisphosphate, and fructose 2.6-bisphosphate, respectively, decrease the reactivity of the first class of thiol groups but not the total number of the accessible cysteins. This result is discussed with regard to conformational changes of the enzyme after binding of the sugar phosphates. PMID- 2977100 TI - [The long-term effects of motivation and plaque control on blind children]. PMID- 2977101 TI - Mating type response in Neurospora crassa. Early and transient changes in the patterns of protein synthesis in sexually stimulated mycelia. AB - 1. Pulse labeling with [35S]-methionine, one-dimensional SDS-polyacrylamide gel electrophoresis and fluorography were used to study the pattern of protein synthesis in Neurospora crassa mycelia undergoing sexual development. 2. Contact of sexually-competent mycelium with cells of the opposite mating type elicited a rapid and transient increase in the synthesis of two predominant proteins of 58 kDa and 40 kDa localized in the cytosol fraction. 3. Marked changes in the pattern of protein synthesis were also observed in the 12,000 g particulate fraction, predominantly mitochondrial, where the synthesis of a 34 kDa polypeptide was most prominent among others. 4. Poly(A)+ RNA extracted from mycelia 2 h after sexual stimulation supported the in vitro synthesis of the 58 kDa and 40 kDa major polypeptides synthesized in vivo. 5. No differences were observed in the pattern of protein synthesis of treated cultures and controls 24 h after the sexual stimulus. PMID- 2977102 TI - Calcium transport through the placenta. AB - The calcium content of the foetus increases exponentially during gestation. The transfer of calcium from the mother to the foetus is believed to occur via an active mechanism, based on observations that the calcium concentration is higher in foetal than in maternal blood, and that perfusion in situ of the umbilical artery of the placenta results in a net increase in the calcium concentration of the perfusate even when the calcium content of the perfusate is threefold higher than in the maternal blood. The calcium concentration in the foetal blood is mainly regulated by foetal parathyroid hormone and the plasma concentration of 1.25-(OH)2 vitamin D3. Several experiments suggest that this vitamin D metabolite plays a key role in calcium transport through the syncytial cell. A vitamin D dependent calcium binding protein has been detected in the placenta, which might intervene in the hormonal control of this transport. The cellular mechanisms of calcium transport through the placenta are not clearly defined. An ATP-dependent calcium uptake by microsomal membrane vesicles from human placenta has been described by several investigators. This calcium transport is saturable, magnesium dependent, and displays Michaelis-Menten kinetics with a Km of approximately 70 nM ionized calcium. Two Ca2+ ATPases with low and high affinities for calcium are described. Since there are discrepancies concerning the exact localization of these enzymes in the syncytial cell, it has not been established whether they intervene in calcium transport through the placenta.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977103 TI - Estimation of blood concentration of drugs after topical application from in vitro skin permeation data. I. Prediction by convolution and confirmation by deconvolution. PMID- 2977104 TI - Amino acid sequence of rabbit factor H of complement. Purification of peptides produced by cyanogen bromide cleavage. PMID- 2977105 TI - Calcium-binding protein regucalcin is an activator of (Ca2+-Mg2+)-adenosine triphosphatase in the plasma membranes of rat liver. PMID- 2977106 TI - Genetic evidence favouring cytotoxic T cell forbidden clones as the cause of insulin-dependent diabetes mellitus. AB - Recent evidence indicates that immunoglobulin light chain variable region genes are genetic determinants for Graves' disease, which is caused by autoantibodies which stimulate the thyroid gland. We have tested whether germline immunoglobulin kappa light chain variable region (V kappa) genes contribute to the genetic predisposition for IDDM. Status for the kappa light chain constant region (C kappa) allotype, Km(1), was determined in members of suitable multiplex IDDM families. Such families had two or more siblings with IDDM, together with one parent negative for Km(1) and the other heterozygous, so that each sibling had a 50% chance of receiving the kappa light chain marker. Twenty-one families of this type were found. Of the siblings, disregarding disease status, 31 were concordant with the diabetic proband for Km(1) and 29 were discordant, this close approximation to equality confirming the validity of the methodology. Of the diabetic siblings of the probands, 14 were concordant and 15 discordant. Of the non-diabetic siblings, 17 were concordant and 14 discordant. This similarity shows that V kappa genes, which are closely linked to C kappa genes, are not genetic determinants for IDDM. The contrast with Graves' disease favours the possibility that the islet beta cell destruction of IDDM is mediated by forbidden clones of cytotoxic T cells, rather than of B cells. PMID- 2977107 TI - Genetic and environmental control of diabetes induction by multi-dose streptozotocin in two BALB/c substrains. AB - BALB/cJ male mice were resistant and BALB/cByJ males were susceptible to induction of diabetes by multi-dose streptozotocin (MSz). Although both closely related BALB/c substrains expressed H-2d haplotype, they could be differentiated by allelic differences at three genetic loci [Qa-2 (Chr 17), Bcd-1 (Chr 5), and Afr-1]. (BALB/cJ X BALB/cByJ)F1 males inherited the BALB/cJ resistance phenotype in a dominant fashion, thereby eliminating the BALB/cJ-expressed Afr-1b (recessive) allele as the susceptibility locus. Backcross of F1 mice to the susceptible BALB/cByJ strain produced a 1:1 segregation of susceptible and resistant (F1-like) phenotypes, suggesting that susceptibility was controlled by a single recessive gene. No linkage was found between the putative susceptibility gene and the mutant BALB/cByJ Qa-2,3 gene linked to the H-2 complex or with the mutant Bcd-1c allele. Since the resistant F1 males expressed low levels of androgen-dependent mouse urinary protein characteristic of the resistant BALB/cJ parental strain, the possibility was discussed that the alleles controlling sensitivity to MSz also controlled tissue sensitivity to endogenous androgens. An environmental effect on phenotype expression was indicated when BALB/cByJ males obtained from a colony free of pneumonia virus of mice (PVM) showed an attenuated rate of response to hyperglycemia induction in comparison to males obtained previously from an enzootically infected colony. PMID- 2977108 TI - Teicoplanin: an investigational glycopeptide antibiotic. AB - The chemistry, mechanism of action, antimicrobial spectrum, pharmacokinetics, adverse effects, and clinical uses of teicoplanin are reviewed. Teicoplanin, a novel glycopeptide that is similar to vancomycin, was isolated in the mid-1970s. A fermentation product of Actinoplanes teicomyceticus, teicoplanin is a structurally complex compound made up of six fatty-acid components attached to a common aglycone. Teicoplanin's mechanism of action, like that of vancomycin, is inhibition of cell-wall biosynthesis. In vitro activity is comparable to that of vancomycin and includes staphylococci, streptococci, corynebacterium, listeria, and anaerobic cocci. Resistance to teicoplanin has been reported with coagulase negative staphylococci. Teicoplanin is 50 to 100 times more lipophilic than vancomycin. Teicoplanin is poorly absorbed after oral administration but is 90% bioavailable when administered intramuscularly. The drug distributes widely into body tissue and is eliminated primarily renally. Optimal dosing regimens and therapeutic serum drug concentrations have not been well established. Reported adverse effects have included irreversible ototoxicity, allergic reactions with maculopapular rash and eosinophilia, pain at intramuscular injection site, and elevation of aminotransferases. Initial clinical trials have yielded conflicting results in gram-positive bacteremia, endocarditis, osteomyelitis, and soft-tissue infections. Teicoplanin has shown promise in surgical and dental prophylaxis. Comparative trials with vancomycin and other antimicrobial agents must be completed before teicoplanin's role as a therapeutic agent in the treatment of systemic gram-positive infections is defined. PMID- 2977109 TI - Use of ipratropium bromide in obstructive lung disease. AB - The chemistry, pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosage of ipratropium bromide are reviewed. Ipratropium bromide, a synthetic quaternary isopropyl derivative of atropine, interrupts vagally mediated bronchoconstriction by inhibiting the cyclic guanosine 3',5' monophosphate system at parasympathetic nerve endings. Ipratropium bromide is poorly absorbed after oral and inhaled administration but diffuses rapidly into tissue after i.v. or i.m. administration. The elimination half-life is 3.2-3.8 hours. After inhalation, the drug is eliminated in the urine and feces. The bronchodilatory effect of ipratropium bromide in stable chronic obstructive pulmonary disease appears to be comparable, and may be superior, to that of the beta-sympathomimetic agents. In acute exacerbations, ipratropium bromide is useful but may not be the preferred agent because of a delayed onset of action (within 15 minutes; mean dose-dependent duration of effect, three to five hours). Combination therapy with other bronchodilating drugs has proved useful. Ipratropium bromide may be a useful adjunctive agent in the treatment of asthma. Since the onset of action is delayed, ipratropium bromide should not be used as single-drug therapy in an acute asthmatic exacerbation. Reported adverse effects, including cough, nausea, palpitations, dry mouth, nervousness, gastrointestinal distress, and dizziness, have been mild. The usual dosage is two inhalations (36 micrograms) four times daily, and the maximum number of doses per day should not exceed 12. Although ipratropium bromide is currently indicated only for maintenance therapy in stable chronic bronchitis and emphysema, it may be useful as adjunctive therapy in asthma and in the management of acute exacerbations of chronic bronchitis and asthma. Additional experience in a variety of chronic obstructive pulmonary disorders will help to clarify the role of ipratropium bromide in the treatment of obstructive pulmonary disease. PMID- 2977110 TI - LHRH and beta-endorphin in the hypothalamus of the ram in relation to photoperiod and reproductive activity. AB - The regional distribution and relative concentrations of LHRH and beta-endorphin immunoreactivity in the hypothalamus were compared in sexually active and sexually inactive Soay rams. LHRH was localized principally in the medial basal hypothalamus and median eminence with lower concentrations in the anterior hypothalamus and preoptic area. beta-endorphin immunoreactivity was also found in highest concentrations in these regions but was more widely distributed into the rostral, dorsal and lateral parts of the hypothalamus. LHRH content in the medial basal hypothalamus was significantly lower in the sexually active rams compared to inactive rams while there was no difference in beta-endorphin immunoreactivity content between the two groups. Gel filtration chromatography of hypothalamic extracts revealed that the beta-endorphin immunoreactivity was due to both beta lipotropin and beta-endorphin; the ratio of beta-endorphin: beta-lipotropin tended to be less in the sexually active rams than in the sexually inactive rams. The results are consistent with the hypothesis that beta-endorphin is a physiologically important endogenous opioid involved in the modulation of LHRH secretion. PMID- 2977111 TI - [Role of the practitioner faced with LAV infection]. PMID- 2977112 TI - [Dental care for patients with AIDS]. PMID- 2977113 TI - [Information for patients]. PMID- 2977114 TI - [Students and AIDS. Questionnaire from the Dental Students Association of Strasbourg]. PMID- 2977115 TI - Effect of dosing intervals on efficacy of clarithromycin and erythromycin in mouse infection models. AB - Clarithromycin is a new macrolide with a serum half-life which is at least twice that of erythromycin. In order to select the appropriate dosing intervals, the in vitro post-antibiotic effect (PAE) of clarithromycin and erythromycin were compared and the efficacies of clarithromycin and erythromycin when administered once (q.d.), twice (b.i.d.) and three (t.i.d.) times a day in mouse protection tests were measured. The doses were selected from achievable peak serum concentrations in man and were 18 mg/kg for clarithromycin which gave a peak blood level of 2.0 micrograms/ml and a serum half-life of 0.82 h, and 10 mg/kg for erythromycin which gave a peak blood level of 0.9 microgram/ml and a serum half-life of 0.46 h. The PAE of clarithromycin for Staph. aureus is 6.25 h compared to 2.35 h for erythromycin. Clarithromycin was effective against Staph. aureus, Strep. pyogenes and Strep. pneumoniae when administered q.d., b.i.d. and t.i.d. and there was no difference in the cumulative mortalities in the three treatment groups. For erythromycin, there was also no difference in the survival of Strep. pyogenes and Strep. pneumoniae-infected mice, but when tested against Staph. aureus it was more effective when administered t.i.d. than b.i.d., and b.i.d. was more effective than q.d. Clarithromycin administered q.d. was more effective than erythromycin administered t.i.d. against Staph. aureus. PMID- 2977116 TI - Effects of cadmium ingestion on blood pressure and ventricular mass in rabbits. AB - Ingestion of cadmium (Cd) acetate in deionized drinking water (1 ppm) in rabbits resulted in the development of hypertension and increased left ventricular mass similar to what was observed in experimental renal hypertension (Grollman technique). Regardless of the approach, mean arterial pressures (MAP) of at least 50 mm Hg greater than those of controls developed over a 34-day period. Increased left ventricular mass relative to body weight was found in both hypertensive groups. However, only an increase in renal mass was observed in unilaterally nephrectomized animals. The increased mass of hearts and kidneys that was detected after an approximately 1-month period attests to the severity of these forms of experimental hypertension. These results demonstrate that the ingestion of drinking water contaminated with Cd can cause hypertension and an increase in left ventricular mass over a short time period in rabbits. PMID- 2977117 TI - Serotonin and platelet aggregation in patients with essential hypertension compared with a normotensive control group. PMID- 2977118 TI - The D-1 dopamine receptor partial agonist, CY 208-243, exhibits antiparkinsonian activity in the MPTP-treated marmoset. AB - Administration of L-DOPA plus carbidopa, or the D-2 agonist (+)-PHNO, to MPTP treated common marmosets caused motor hyperactivity and a reversal of the parkinsonian syndrome. In contrast, administration of the putative D-1 agonist SKF 38393 was without effect on movement or motor disability. The subsequent administration of another putative selective D-1 partial agonist CY 208-243 produced a dose-related improvement in motor activity and reversal of parkinsonian motor deficits in MPTP-treated animals. The effect of CY 208-243 was inhibited by pretreatment with the D-1 antagonist SCH 23390 and, to a lesser extent, by the D-2 antagonist sulpiride. In another group of normal drug naive marmosets, the administration of CY 208-243 produced only a small increase in motor activity. Following treatment with MPTP and without other drug administration, administration of CY 208-243 produced a marked reversal of motor deficits and locomotor hyperactivity. Thus, CY 208-243, suggested to be a partial D-1 agonist exhibits antiparkinsonian activity in MPTP-treated marmosets which does not require prior or concurrent exposure to D-2 agonists. PMID- 2977119 TI - Therapeutic choices for rheumatic disorders. AB - Of concern to physicians specializing in the elderly is the different profile of drug metabolism and toxicity in the elderly. Older patients have differences in metabolism of a number of drugs, as well as in altered immunologic responses to these agents. The advantages and disadvantages of the agents used most frequently to treat elderly rheumatic patients will be reviewed. For example, effects of various nonsteroidal antiinflammatory drugs (NSAIDs) regarding efficacy and adverse gastrointestinal, hepatic, and renal effects, and their different pharmacokinetic profiles can play a part in choosing appropriate therapy for an individual patient. Etodolac, a newer NSAID, is a promising addition to the available choices for elderly, as well as younger, patients. Etodolac has been shown to be clinically effective with a low incidence of adverse effects. It has a half-life of approximately 7 h in older patients, and about 75% of the drug is excreted in the urine mainly as metabolites. It is highly protein-bound. PMID- 2977120 TI - Relaxation of rabbit middle cerebral arteries in vitro by H1 histaminergic agonists is inhibited by indomethacin and tranylcypromine. AB - The H1-histaminergic agonists 2-pyridylethylamine (2-PEA) and 2-methylhistamine relaxed potassium-constricted, perfused, rabbit middle cerebral arteries at low concentrations (3 x 10(-11) to 3 x 10(-8) M) and constricted them at high concentrations (3 x 10(-7) to 3 x 10(-4) M). The relaxation and the contraction were not antagonized by propranolol (up to 3 x 10(-6) M) given 30 min before, suggesting that beta-adrenergic mechanisms were not involved. When 2-PEA was tested on arteries constricted with uridine triphosphate (UTP), similar results were obtained. In the UTP-constricted arteries, the 2-PEA-induced responses were competitively antagonized by 3 x 10(-9) M mepyramine. Together with previous work (Ea Kim et al., 1986), these results are compatible with the hypothesis that H1 receptors were responsible for both the relaxation and the contraction observed. When either indomethacin (10(-8), 3 x 10(-7), or 10(-5) M), dexamethasone (10(-5) M), or tranylcypromine (10(-5) or 10(-4) M) were tested on the response to 2-PEA or 2-methylhistamine, these inhibitors suppressed the relaxation or reversed it to a contraction. Furthermore, they potentiated the contraction induced by these agonists. These results favour the hypothesis that the H1-mediated relaxation in rabbit cerebral arteries may in part involve the release of prostaglandins, especially prostacyclin. The participation of such a prostanoid in histaminergic relaxation seems exclusively an H1-mediated mechanism, since the relaxation induced by the H2-agonist dimaprit (in the presence of mepyramine) was not antagonized by either indomethacin (3 x 10(-7) M) or tranylcypromine (10(-4) M). PMID- 2977121 TI - [Localization of atrial natriuretic peptide in the heart. Immunohistochemical and northern blot analyses]. AB - Atrial natriuretic peptide (ANP) which is secreted from atrial muscle has been shown to produce relaxation of vascular smooth muscle, anti-hypertensive effects, and natriuresis, and to implicate renin-angiotensin-aldosterone network. In human, ANP (hANP) exists as three subtypes: alpha-, beta-, gamma-hANP which have molecular weight of 3,000, 6,000 and 13,000, respectively. In human tissue hANP has been shown to be present not only in heart atria, but also submandibular gland. In addition, it has been reported that in some conditions of heart disease hANP can be found in the ventricles. However, the exact distribution outside of the atrial system has not been definitely established. In our studies, we attempted by using immunohistochemical methods on human and dog hearts to answer this question. In addition, by using nucleic acid probes we have investigated possible areas where hANP may be synthesized. In the initial studies specific antibody to hANP was prepared in rabbits by immunization with synthetic alpha hANP coupled to porcein thyroglobulin. The specificity of this antibody was confirmed by Western Immunoblotting. Immunoperoxidase staining demonstrated hANP in His-bundle and major branching bundles as well as atrial wall and AV-node of both human and dog hearts. Ventricular muscle cells outside of the conduction system did not contain hANP. The possibility of non-specific staining by antibody to thyroglobulin was muled out as antibody to thyroglobulin alone never showed positive staining. However, staining of the atrial muscles was always granular in the perinuclear areas, while that of the conduction system was usually diffuse.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977122 TI - Membrane potential and activities of certain membrane-bound enzymes of polymorphonuclear leukocytes in essential hypertension. PMID- 2977123 TI - Astrocytes as eicosanoid-producing cells. AB - A variety of prostaglandins and leukotrienes, together with thromboxane and prostacyclin metabolites, can be detected in central nervous tissues and in cerebrospinal fluid. Defined cultures of astrocytes have revealed these cells to be a major source of eicosanoids. In common with other eicosanoid-producing cells, agents such as calcium ionophores and phorbol esters are potent stimuli for promoting release. While in other tissues agonists for receptors linked to calcium mobilisation prompt eicosanoid release, this does not seem to be the case in astrocytes, though a range of such receptors are present. The notable exceptions to this observation are adenosine triphosphate and adenosine diphosphate, presumably acting through P2 purinergic receptors. Many cell types in the CNS are targets for eicosanoids, possessing receptors linked to adenylate cyclase or phospholipase C. An appreciation of the functional significance of activation of these receptors is just now beginning. Eicosanoids have effects in the CNS that involve not only the vascular supply but also synaptic modulation and immune regulation. PMID- 2977124 TI - Distribution of orthogonal arrays of particles in the Muller cell membrane of the mouse retina. AB - In the present study we investigated the Muller cell membrane of the mouse retina by freeze-fracturing. The mouse retina is vascularized and the vessels running outside the nerve fiber layer are completely encased by Muller cell endfeet. Orthogonal arrays of particles (OAP) reside in all membrane areas of the Muller cells. The paravitreous as well as the pericapillary endfeet reveal a considerably higher density of OAP than the nonendfoot membranes including the perikaryal ones. This is in contrast to the Muller cell membrane of the rabbit retina studied previously (Wolburg and Berg: Neurosci, Lett., 82:273-277, 1987). There we found a completely different distribution of OAP; practically all OAP reside in the endfoot membrane facing the vitreous body. The nonendfoot and perikaryal membranes were devoid of OAP. The OAP distribution in both species corresponds roughly to the distribution of the K+ conductances measured by Newman (J. Neurosci., 7:2423-2432, 1987). The putative relationship between OAP and K+ channels, including functional aspects, is discussed. PMID- 2977125 TI - Cytoplasmic membrane elaborations in oligodendrocytes during myelination of spinal motoneuron axons. AB - The ultrastructure of paranodal oligodendroglial cytoplasm, which is located in proximity to the forming myelin sheath, was studied during maturation of spinal motoneuron axons in rat. At 8 days postnatal, the paranodal oligodendroglial loops contain a network of membrane-bound tubulovesicular elements. These membrane elaborations are most common in oligodendroglial loops attached to the outermost layers of the myelin sheath, i.e., paranodal loops closest to the nodal gap. The number of oligodendroglial cytoplasmic profiles per paranodal loop falls over the course of five to ten sequential paranodal loops, and these profiles are nearly absent in paranodal oligodendroglial cytoplasm located distant from the nodal gap. Oligodendrocytes in spinal cords of 14- and 20-day-old rats and of adult rats did not exhibit networks of tubulovesicular profiles. The appearance of these membrane organelles within oligodendroglial cytoplasm during myelin maturation suggests increased membrane turnover within paranodal cytoplasm located adjacent to the axon that is being myelinated. Membrane turnover within oligodendrocytes may reflect axonal modulation of glial function during myelination. PMID- 2977126 TI - [Disability prevention in Hansen's disease: reality at a health service]. AB - In Hansen's disease, the third part of advanced incidences or non treated cases exhibit some patterns of physical incapacity, besides the prejudice and beliefs generated by deformities-which the patients may present if they do not get a correct treatment. Which basis on this problematical question of the disease a group of professionals composed of medical doctors, nurses, physical therapist, occupational therapist and social worker made a study of the incapacities of the Hansen's disease patients through a detailed physical exam. This study intended to identify sensory and motor injuries of the face, upper and lower limbs. This study also develops a research about the necessity to systematize the assistance given in a public health center. PMID- 2977127 TI - [Leprous symmetric keratoderma palmaris et plantaris. Apropos of 2 clinical cases]. AB - Two cases of "Keratoderma palmaris et plantaris symmetric leprous" are described, which rarely occur in the field of acrokeratosis, contracted by infection. PMID- 2977128 TI - Liquid chromatographic determination of zearalenone and alpha- and beta zearalenols in milk. AB - Previous research has demonstrated transmission of zearalenone and alpha- and beta-zearalenols into the milk of cows and other animals. Since human intake of zearalenone and its metabolites via milk is an unknown factor in risk assessment of zearalenone and because appropriate methodology for their determination in milk is not available, a rapid and sensitive analytical method has been developed. Essentially, the method includes extraction with basic acetonitrile, acidification, partition into methylene chloride on a hydrophilic matrix, cleanup on an aminopropyl solid phase extraction column, and reverse-phase liquid chromatography with fluorescence detection. Recoveries from milk averaged 84% for zearalenone, 93% for alpha-zearalenol, and 90% for beta-zearalenol at spiking levels of 0.5 to 20 ng/mL. As little as 0.2 ng/mL of zearalenone and alpha zearalenol and 2 ng/mL of beta-zearalenol can be detected in milk. These 3 compounds are stable in refrigerated milk for at least 2 weeks and in milk brought to boiling. Enzymes (beta-glucuronidase and aryl sulfatase) may be added to milk prior to extraction to hydrolyze any conjugates. PMID- 2977130 TI - The amino acid sequence of ribonuclease N1, a guanine-specific ribonuclease from the fungus Neurospora crassa. AB - The complete amino acid sequence of ribonuclease N1 (RNase N1), a guanine specific ribonuclease from a fungus, Neurospora crassa, was determined by conventional protein sequencing, using peptide fragments obtained by tryptic digestion of cyanogen bromide-treated RNase N1 and by Staphylococcus aureus V8 protease digestion of heat-denatured RNase N1. The results showed that the protein is composed of a single polypeptide chain of 104 amino acid residues cross-linked by two disulfide bonds and has a molecular weight of 11,174: (sequence; see text) (Disulfide bonds: C2-C10, C6-C103) The amino acid sequence was homologous with those of RNase T1 (65% identity) and related microbial RNases. PMID- 2977129 TI - The conformation of alpha-human atrial natriuretic polypeptide in solution. AB - The three-dimensional structure of alpha-human ANP in solution was determined through the combined use of nuclear magnetic resonance spectroscopy and distance geometry. The results are based on distance constraints determined by nuclear Overhauser effect measurements and one disulfide bond. The structure is as follows. Three separate regions, which are Ser1-Cys7, Arg11-Ile15, and Gln18 Tyr28 each have some ordered structure. The remaining parts in the sequences of Gly9-Gly10 and Gly16-Ala17 act as hinges. And the C-terminal part is folded back toward the cyclic moiety. The conformation of alpha-hANP reported here is expected to give a better understanding of the relationships between its biological activities and three-dimensional structure. PMID- 2977131 TI - The different roles of two distinct Fc gamma receptors on guinea pig macrophages in the phagocytosis of sensitized sheep erythrocytes. AB - The functional roles of two distinct types of Fc gamma receptors (Fc gamma 1/gamma 2R specific for both IgG1 and IgG2, and Fc gamma 2R specific for IgG2 alone) on the surface of guinea pig macrophages in the phagocytosis of sensitized sheep erythrocytes (EA) were investigated by the use of two Fab's of monoclonal anti-Fc gamma 1/gamma 2R and anti-Fc gamma 2R antibodies. The binding and subsequent ingestion of IgG1 antibody-sensitized erythrocytes (EA gamma 1) by macrophages were completely inhibited by anti-Fc gamma 1/gamma 2R Fab', indicating that the reactions are mediated only by Fc gamma 1/gamma 2R. On the other hand, the binding and subsequent ingestion of IgG2 antibody-sensitized erythrocytes (EA gamma 2) were substantially inhibited by anti-Fc gamma 2R Fab', but not by anti-Fc gamma 1/gamma 2R Fab'. The inhibitory activities of anti-Fc gamma 2R Fab' were dependent upon the amount of IgG2 antibody bound on erythrocytes; increasing the amount of bound IgG2 antibody from 0.15 to 0.91 micrograms/2 X 10(8) erythrocytes resulted in a decrease in the inhibition of binding of EA gamma 2 by anti-Fc gamma 2R Fab' from 50 to 0%, and also a decrease in the inhibition of ingestion of EA gamma 2 from 100 to 50%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977132 TI - The yeast peptide elongation factor 3 (EF-3) carries an active site for ATP hydrolysis which can interact with various nucleoside triphosphates in the absence of ribosomes. AB - ATP (GTP) hydrolysis was clearly demonstrated by using at most 16 pmol of yeast peptide elongation factor 3 (EF-3) in the absence of ribosomes. However, the highly active yeast ribosomes (up to 48 pmol) displayed virtually no ATPase (or GTPase) activity in the absence of EF-3. Several lines of evidence indicated that both the catalytic and binding sites of the ATPase reside in the elongation factor itself, not on the ribosomes. The patterns of protection by various nucleoside triphosphates against tryptic digestion of EF-3, reflecting the wide substrate specificity of the ATPase, confirmed that the active center of the endogenous ATPase is located on the factor itself and not on contaminants. The intrinsic activity was stimulated up to two orders of magnitude by the presence of the yeast ribosomes fully active in polyphenylalanine synthesis. The activation was achieved by enhancing the catalytic activity (kcat) to a much greater extent than the binding affinity (Km). On the other hand, the ribosome activated ATPase activity was revealed to inherit its wide substrate specificity from the intrinsic property of EF-3, which shows an affinity to various XTPs, including pyrimidine- and purine-nucleoside triphosphates, irrespective of 2' hydroxylation of the sugar moiety. From experiments on protection against tryptic digestion, we determined that intricate conformational changes of the factor molecule occur upon interaction with the substrate XTP and ribosomes. PMID- 2977133 TI - Oxygen exchange during the acto-subfragment-1 ATPase reaction: evidence for the two-route mechanism of the actomyosin ATPase reaction. AB - The oxygen exchange occurring during the acto-S-1 ATPase reaction was analyzed based on the distribution of 18O-labeled species of P1 using [gamma-18O]ATP as a substrate. Evidence was found for the two-route mechanism in which ATP is hydrolyzed via the dissociation of acto-S-1 into F-actin and the S-1-phosphate ADP complex, S-1PADP, and their recombination, and also hydrolyzed without the dissociation of acto-S-1 (Inoue, A., Shigekawa, M., & Tonomura, Y. (1973) J. Biochem. 74, 923-934; Inoue, A., Ikebe, M., & Tonomura, Y. (1980) J. Biochem. 88, 1663-1677). When ATP was mainly hydrolyzed without the dissociation of acto-S-1, the extent of oxygen exchange was low. When ATP was hydrolyzed by both routes, the distribution of product P1 with 3, 2, 1, and 0 18O atoms showed a mixture resulting from low and high oxygen exchange. The rate of ATPase without the dissociation of acto-S-1 can be estimated from the rate of the overall reaction (v), the rate of recombination of S-1PADP with F-actin (vr), and the extent of dissociation of acto-S-1 (a). The distribution of the P1 species measured was almost equal to that calculated from the ratio of ATP hydrolysis via the two pathways as avr and v-avr, respectively. This result indicates that the rates of the dissociation of acto-S-1PADP into S-1PADP and F-actin and their recombination are much lower than the rate of decomposition of the acto-S-1PADP complex into acto-S-1 + ADP + Pi. PMID- 2977134 TI - Synthesis of recombinant human single-chain urokinase-type plasminogen activator variants resistant to plasmin and thrombin. AB - Single-chain urokinase-type plasminogen activator (scu-PA), a potential therapeutic reagent for thrombosis, is activated in plasma by plasmin. The activated enzyme is further digested by plasmin to generate low-molecular-weight urokinase (LMW-UK), which has no affinity for fibrin. To circumvent this dual effect of plasmin, we synthesized in Escherichia coli a variant of scu-PA, which is not converted to LMW-UK on treatment with plasmin. In another variant, the activation cleavage site was modified such that activation by plasmin was slowed down and that inactivation by thrombin was greatly diminished. The combination of these variants may be applicable as an effective thrombolytic reagent for clinical use. PMID- 2977135 TI - Cyclosporin A-induced reversible cortical blindness. AB - Despite the occurrence of serious side effects, the use of cyclosporin A after organ transplantation has increased because of its ability to effectively suppress allograft rejection. Its use in the treatment of ophthalmic disease has also recently increased. Central nervous system toxicity due to cyclosporin A is a significant but apparently clinically reversible side effect. A liver transplant patient in whom cortical blindness from profound neurotoxicity was the initial presentation is described. Neurologic abnormalities, including cortical blindness, resolved completely after discontinuation of cyclosporin A. However, pathologic studies performed 8.5 months after the initial transplant revealed residual central nervous system demyelination. PMID- 2977136 TI - Transient cortical blindness with occipital lobe epilepsy. AB - Three youths with occipital seizures and intermittent visual loss are presented. All three had occipital epileptiform discharges. Visual loss occurred during seizures and improved with anticonvulsant medication. In the absence of an underlying structural lesion of the occipital lobe, occipital seizures in children often respond to antiepileptic medication and may spontaneously resolve. As occipital seizures are often associated with headaches and sometimes with vomiting, differentiating them from basilar migraine may be difficult. The possibility of ongoing seizure activity should be considered in a patient who presents with acute visual loss without an obvious cause. PMID- 2977137 TI - Does sporadic Leber's disease exist? AB - This study gives some illustrative case reports of the difficulties in the diagnosis of Leber's hereditary optic neuroretinopathy. It underlies the importance of careful family history and search for peripapillary microangiopathy in the maternal relatives of patients suspected to suffer from Leber's disease. The article casts doubt on the existence of so-called sporadic Leber's disease. PMID- 2977138 TI - Optic nerve ultrastructure following amiodarone therapy. AB - Amiodarone has been implicated in the pathogenesis of optic neuropathy in several cases. However, that relationship is unclear, as subjects placed on amiodarone represent a high-risk group for various vasoocclusive accidents. In order to investigate the effect of amiodarone on the optic nerve, we examined histopathologically sections of the retrobulbar optic nerve obtained from an asymptomatic subject taking amiodarone. Lamellar inclusions were selectively found in the large axons. Amiodarone may have a chronic neurotoxic effect on the optic nerve via a drug-induced lipidosis. This neurotoxicity may be related to some of the acute forms of optic neuropathy described in the literature. PMID- 2977139 TI - Recurrent anterior ischemic optic neuropathy in young adults. AB - Anterior ischemic optic neuropathy (AION) is primarily a disease of middle and late life. When it affects the young patient, AION is often due to a specific underlying disease process predisposing to vasculopathy. Reported here are two patients with AION in whom no evidence of associated disease was found. The tendency for recurrent attacks of disc ischemia distinguishes this clinical entity from the more common idiopathic form of AION of older individuals. Hypotheses regarding possible etiology are discussed. PMID- 2977140 TI - Melkersson's syndrome associated with syphilis. AB - The onset of the Melkersson-Rosenthal syndrome, manifested by bifacial paralysis and severe edema of the upper lip in a 25-year-old man, was followed shortly by signs of secondary syphilis. PMID- 2977141 TI - Sneddon's disease presenting with visual loss and dementia. AB - A 51-year-old woman with Sneddon's disease presented with transient right hemifield loss of vision and transient right-sided weakness. Over the preceding decade she had experienced a slow decline in mental function. She also had hypertension, migraine, and a mixed seizure disorder. She had skin changes typical for generalized livedo reticularis but she did not have Raynaud's phenomenon or winter ulcerations. Her disease was not understood until the stroke related symptoms were associated with the skin abnormalities. We review the neuro ophthalmic manifestations of Sneddon's disease and add data from our case to the growing body of fact that suggests that Sneddon's disease may be an immunologically mediated vasculopathy. PMID- 2977143 TI - Magnifying loupe. PMID- 2977142 TI - Trochlear nerve palsy following minor head trauma. A sign of structural disorder. AB - Trauma-induced superior oblique palsy usually results from contusion or avulsion of the trochlear nerve or from decompensation of a congenital trochlear nerve palsy. Severe craniocerebral trauma is often associated with the former mechanism, whereas more minor closed-head injuries can decompensate a congenital phoria. We report a patient who developed an isolated trochlear nerve palsy following minor head trauma. Investigation revealed an unsuspected tentorial vascular malformation that was compressing the trochlear nerve in its subarachnoid course. In the absence of other features (e.g., documentation of old head tilt, large vertical fusion amplitudes) that support decompensation of a congenital phoria, compressive lesions should be sought in cases of fourth cranial nerve palsies that follow minor head trauma. PMID- 2977144 TI - Shunt-dependent hydrocephalus. PMID- 2977145 TI - Recurrent diplopia after vestibular nerve section. PMID- 2977146 TI - Imaging techniques in optic nerve evulsion. PMID- 2977147 TI - [T cell antigen receptor gene rearrangement in refractory anemia with excess of blasts (RAEB)]. PMID- 2977148 TI - [A case of postoperative craniopharyngioma with hypernatremia. The mechanisms of ADH and atrial natriuretic peptide (ANP) releases]. PMID- 2977149 TI - Frontal-plane QRS axis revisited: accuracy of current approximations and reappraisal of their merit in the diagnosis of right ventricular hypertrophy. AB - The frontal-plane mean QRS vector orientation (AQRSxy)--the so-called electrical axis--is an ECG feature commonly used for the diagnosis of right ventricular hypertrophy and is correctly measured by calculating the areas subtended by QRS deflections in two different leads. To overcome the drawbacks of doing this by hand, two alleged approximations of AQRSxy have become popular and are in current use: one is based on the measurement of QRS component wave peak amplitudes and the other on the estimation of the half-area vector of the frontal plane loop. The values obtained with the correct and the two more practical methods are compared and their diagnostic efficiency is assessed by means of a procedure for ECG criteria optimization based on the receiver operating characteristics (ROC) curve analyzed in terms of information theory. The authors conclude that the two more popular methods for AQRSxy determination provide similar values that, although correlated with the true measure of the parameter are statistically different from it. On the other hand, the diagnostic efficiency of AQRSxy alone, regardless of the method by which it is computed, is only as good as, if not bettered by, other much more easily measurable frontal-plane parameters (ie, left to rightward forces amplitude ratio in adults and rightward forces amplitude in pediatric patients). PMID- 2977150 TI - Characteristics of M-GTFI, a new inhibitor of Streptococcus mutans glucosyltransferase. AB - M-GTFI, originally screened as an inhibitor of Streptococcus mutans glucosyltransferase, strongly inhibited alpha-glucosidase, in a non-competitive manner especially when the synthetic substrate p-nitrophenyl-alpha-D glucopyranoside was used. It also inhibited beta-glucosidase, beta-amylase and, to a lesser extent, beta-glucuronidase. The inhibitor was stable in neutral and alkaline pH ranges and dependency of the inhibition on pH and temperature was not observed. Some proteinases and polysaccharides-hydrolyzing enzymes as well as human saliva did not inactivate the inhibitor. There was a correlation between the release of sulfate anions from the inhibitor molecule on incubation with HCl (0.2 N) at 100 degrees C and loss of inhibitory properties of the molecule. It is suggested that the presence of sulfate ester linkages in the inhibitor molecule play an important role in the inhibition process. PMID- 2977151 TI - Mechanisms behind myocardial depression in rat renal hypertension. AB - Cardiac function in rat renal hypertension has been shown repeatedly to be depressed. The reason for this impairment has not been fully understood, although cardiac hypertrophy and increased collagen content have been claimed as possible causes. To further unravel the mechanisms underlying the deterioration of left ventricular function in rat renal hypertension, we investigated cardiac function, myocardial morphology, myosin iso-enzymes, plasma renin activity and levels of myocardial high-energy compounds in hearts from rats exposed to renal and reversed renal hypertension. Maximal cardiac function was unaltered in hearts exposed to antihypertensive therapy from the time of renal artery clipping compared with untreated hypertensive rats. The observed alterations of iso-enzyme pattern, plasma renin activity levels and myocardial morphology among the groups showed to be of less importance with respect to cardiac performance. Together with previous results from our laboratory, the present findings suggest that some negative inotropic agent of renal or non-renal origin is released during two kidney, one clip renal hypertension, which counteracts the enhanced left ventricular performance induced by cardiac hypertrophy. PMID- 2977152 TI - Cardiac hypertrophy and characteristic impedance in spontaneously hypertensive rats. AB - Characteristic impedance and systemic arterial compliance were evaluated in 12 anaesthetized spontaneously hypertensive rats (SHR) from analysis of phasic records of pressure and flow in the ascending aorta. Cardiac hypertrophy expressed as the ratio of the left ventricular weight to the body weight was not correlated with arterial blood pressure (systolic, diastolic or mean). In contrast, there was a significant relationship between the left ventricular weight to body weight ratio and the characteristic impedance, suggesting that increased arterial stiffness contributes to left ventricular hypertrophy in SHR. PMID- 2977153 TI - Left ventricular hypertrophy and ventricular dysrhythmic risk in hypertensive patients: evaluation by programmed electrical stimulation. AB - Left ventricular hypertrophy in hypertensive patients is associated with an increased prevalence of ventricular arrhythmias. Twelve patients with left ventricular hypertrophy assessed by M-mode echocardiography and 12 without left ventricular hypertrophy underwent an electrophysiological study with programmed electrical stimulation. The patients with left ventricular hypertrophy had a prolonged infranodal conduction time which correlated closely with left ventricular mass (r = 0.71; P less than 0.001). Programmed electrical stimulation initiated more intraventricular re-entry and unsustained ventricular tachycardia in the group with left ventricular hypertrophy than in the control group, although sustained ventricular tachycardia was never induced. We conclude that ventricular vulnerability is increased in hypertensive patients with left ventricular hypertrophy, especially in those who show electrocardiographic evidence of left ventricular hypertrophy. PMID- 2977154 TI - The induction of cardiac hypertrophy by catecholamines can be dissociated from their inotropic effect. AB - In this study, ornithine decarboxylase, the first and rate-limiting enzyme of the polyamine pathway, was used as a marker of the very early stages of cardiac hypertrophy. Our data show that ornithine decarboxylase is independently regulated by ventricular wall stress and adrenergic receptors, and support the theory that catecholamines have a pressure-independent trophic effect. The infusion of beta 2-adrenergic agonist, terbutaline, at 10(-7) mol/l, did not affect the cardiac performance of isovolumic perfused rat hearts. Despite the lack of functional changes, ornithine decarboxylase activity, as assayed in vitro by 14C-ornithine decarboxylation, was markedly increased in both left and right ventricles. Further investigation showed that ornithine decarboxylase stimulation by catecholamines, unlike ornithine decarboxylase basal activity, was dependent on the extracellular calcium level. Furthermore, passive increases in ventricular wall stress in non-beating hearts also increased left ventricular ornithine decarboxylase activity. PMID- 2977155 TI - Soluble factor from the hypertrophied left ventricle of dogs in experimental hypertension: ability to stimulate protein metabolism of cultured heart cells. AB - Our study concerned the existence of humoral factor(s) in cardiac hypertrophy arising from experimental perinephritic hypertension in dogs. Hypertension was induced by the method of Page with some modifications. A microassay technique was used on cultured neonatal rat heart cells. Heart extract from the hypertrophied left ventricle of dogs with experimentally induced renal hypertension, but not sham-operated dogs, increased the uptake of 3H-uridine by cultured rat heart cells. At a final heart extract concentration of 5 X 10(-3)% v/v (1-3 micrograms/ml), 3H-uridine was increased by a mean of about 15%, from four experiments. High performance liquid chromatography showed that the heart extract contained at least 16 molecules. Among them, a molecule of approximately 11,200 molecular weight stimulated the uptake of both 3H-uridine and 14C-leucine by cultured rat heart cells. These results indicate that the heart extract from the hypertrophied left ventricle of dogs with experimentally induced renal hypertension contained a factor that might induce and/or modulate myocardial hypertrophy in the model of hypertension. PMID- 2977156 TI - Expression of c-myc proto-oncogene in hearts and cultured smooth muscle cells of spontaneously hypertensive rats. AB - We studied the expression of c-myc proto-oncogene in hearts and cultured aortic smooth muscle cells of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY), in order to investigate the association of the c-myc gene with cardiac hypertrophy and atherosclerosis in SHR. Transcription of the c-myc gene in hearts of SHR was higher than that of WKY at 10 weeks of age, when cardiac hypertrophy had developed in SHR. The c-myc gene expression in cultured aortic smooth muscle cells of SHR, after the addition of serum to the serum-deprived cultures, was higher than that of WKY. These results suggest that the enhanced expression of the c-myc gene in the hearts and cultured aortic smooth muscle cells of SHR may be associated with the growth control of these cells, and may play a role in the development of cardiac hypertrophy and atherosclerotic lesions in SHR. PMID- 2977157 TI - Disparate difference in preload reserve between myocardial hypertrophy due to essential hypertension and hypertrophic cardiomyopathy. AB - To investigate the left ventricular preload reserve in essential hypertensives, the legs-up procedure was performed and the data were compared with those in patients with non-obstructive hypertrophic cardiomyopathy and in normal subjects. Stroke volume index and the following systolic time intervals were measured before and after the legs-up procedure: left ventricular ejection time, pre ejection period and the ratio of pre-ejection period to left ventricular ejection time (PEP/ET). No changes in blood pressure, heart rate or plasma catecholamine levels were noted before and after the legs-up procedure in all groups. A shortening in the pre-ejection period, a prolongation of ejection time, a decrease in the PEP/ET ratio and an increase in the stroke volume index were observed in essential hypertensives and in normal subjects. However, the changes in these parameters were not observed in patient with hypertrophic cardiomyopathy. The legs-up procedure is known to increase left ventricular end diastolic volume. These results suggest that myocardial hypertrophy due to essential hypertension preserves the same preload reserve as in normals, but hypertrophic cardiomyopathy does not. PMID- 2977158 TI - Sodium intake modulates left ventricular hypertrophy in essential hypertension. AB - In order to assess the impact of dietary sodium intake on the degree of left ventricular hypertrophy, we determined posterior wall thickness, relative wall thickness and left ventricular mass by two-dimensionally guided M-mode echocardiography, and related these parameters to sodium excretion over 24 h. There was no restriction on sodium intake. The first cohort comprised 43 subjects (residents of New Orleans) with mild to moderate essential hypertension who had not been treated for at least 4 weeks; in this cohort sodium excretion correlated with posterior wall thickness (r = 0.64, P less than 0.001), relative wall thickness (r = 0.67, P less than 0.001) and left ventricular mass (r = 0.37, P less than 0.02). A stepwise multiple regression analysis confirmed that sodium excretion was a determinant of posterior wall thickness (P less than 0.02) and relative wall thickness (P less than 0.05) independently of age, arterial pressure and body weight. The second cohort comprised 60 white male patients (residents of Bonn) with mild essential hypertension who had never been treated in the past; in this cohort sodium excretion correlated with diastolic diameter (r = 0.36, P less than 0.001) and with left ventricular mass (r = 0.35, P less than 0.001). Sodium excretion and systolic pressure emerged as independent variables (P less than 0.02) for left ventricular mass as evaluated by multiple regression analysis. These results identify dietary sodium intake as an independent powerful determinant of left ventricular hypertrophy in two disparate patient cohorts. Thus, for a similar haemodynamic load, sodium intake might accelerate, and conversely salt restriction mitigate, cardiac structural adaptation in patients with essential hypertension. PMID- 2977159 TI - The effects of vasoactive agents on the proliferation of vascular smooth muscle cells grown in vitro. AB - The present study was designed to investigate the effect of vasoactive agents on cellular proliferation in serially passed cultured vascular smooth muscle cells (VSMC). A substantial reduction in the number of vascular smooth muscle cells was observed with the addition of nifedipine, nicorandil, bunazocine and labetalol compared with that in a control sample. Furthermore, noradrenaline significantly increased the number of vascular smooth muscle cells. In contrast, neither propranolol nor captopril had any effect on number of vascular smooth muscle cells. The cell size, measured as water volume of vascular smooth muscle cells based on the equilibrium distribution of 3-O-(14C-methyl)-D-glucose, did not differ between treatments with the above-mentioned agents. It is suggested that in addition to a known calcium-mediated mechanism, an alpha-receptor-mediated property could be involved in the proliferation of vascular smooth muscle cells and that clinical use of a calcium antagonist or an alpha-blocker might be useful to prevent the hyperproliferation of vascular smooth muscle cells commonly seen in the vascular walls of patients with hypertension. PMID- 2977160 TI - The importance of hypophyseal hormones for structural cardiovascular adaptation in hypertension. AB - The effect of renal artery clipping was tested in three groups of male Sprague Dawley rats: (1) 30 control animals, (2) 30 hypophysectomized animals, and (3) 30 hypophysectomized animals treated with growth hormone and thyroxine. Fifteen rats in each group were clipped and 15 acted as controls. In the first group clipping raised arterial pressure and plasma renin activity. Thirty-five days after clipping, pair-perfused hindquarter preparations at maximal dilation and maximal pressor response were both increased, reflecting, respectively, decreased lumen diameters and increased media thickness in the resistance vessels. Clipping also increased left ventricular weight. Hypophysectomy eliminated the weight gain, and the maximal pressor response and maximal dilation were lower than in the control and treated groups. Hypophysectomy also considerably reduced the rise in blood pressure on clipping and, even more so, the associated structural cardiovascular changes. Replacement therapy with growth hormone and thyroxine almost restored the weight gain and also the structural responses of the heart and vessels to clipping. We conclude that pituitary hormones play an important, probably permissive, part in the development of normal vessel structure and in the adaptation of cardiovascular structure to chronic hypertension. PMID- 2977161 TI - Size and site-dependent heterogeneity of human vascular responses in vitro. AB - Ring segments of splanchnic, peripheral, coronary, pulmonary and uterine conduit arteries obtained during surgery were studied in tissue baths. Resistance arteries dissected from various sites were studied in a myograph. Both conduit and resistance vessels contracted in response to the alpha 1-agonist phenylephrine (10(-7) to 10(-4) mol/l), an effect that was antagonized by the alpha 1-antagonist doxazosin (10(-8) to 10(-6) mol/l). However, the alpha 2 agonists BHT 933 (10(-7) to 10(-4) mol/l) and UK 14304 (10(-7) to 10(-4) mol/l) only contracted the resistance vessels and not the conduit arteries. The response to BHT 933 was competitively antagonized by the alpha 2-antagonist yohimbine (3.10(-8) to 3.10(-7) mol/l) and the magnitude of the contractile response was inversely related to vessel size. Similarly, neuropeptide Y (10(-9) to 10(-6) mol/l) contracted only the resistance vessels, and induced marked tachyphylaxis. Atrial natriuretic peptide (ANP; 10(-8) to 10(-6) mol/l) produced concentration dependent relaxation in all conduit arteries studied, being ineffective in resistance arteries from subcutaneous or omental sites, but relaxed those from renal tissue and skeletal muscle. Calcitonin gene-related peptide (10(-8) to 10( 6) mol/l) and vasoactive intestinal peptide (10(-9) to 10(-6) mol/l) relaxed both conduit and resistance arteries. This response was dependent on the integrity of the endothelium in the systemic conduit but not the resistance vessels. These results indicate that the receptors for adrenergic agonists and vasoactive peptides are varyingly distributed throughout the human vasculature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977162 TI - Expression of atrial natriuretic factor gene in hearts from neonates of spontaneously hypertensive rats and stroke-prone spontaneously hypertensive rats. AB - In order to further elucidate the expression of the atrial natriuretic factor (ANF) gene in spontaneously hypertensive rats (SHR) and the substrain, stroke prone SHR (SHRSP), ANF messenger (m)RNA levels in the atrium and in the ventricle were measured in neonates of SHR and SHRSP and were compared with those in control Wistar-Kyoto rats (WKY). The levels of ANF mRNA in ventricles of the three strains reached approximately 7% of those in atria; however, no significant difference was observed in atrial and ventricular ANF mRNA levels among the three strains. These results demonstrate that the expression of the ANF gene in hearts from neonates of WKY, SHR and SHRSP is similar among the three strains. PMID- 2977163 TI - Ganglionic, spinal cord and hypothalamic atrial natriuretic factor: its distribution, origin and possible role in spontaneously hypertensive rats. AB - We report the occurrence of the atrial natriuretic factor (ANF) prohormone in the hypothalamus, spinal cord and sympathetic ganglia determined by measurement of immunoreactive ANF by two peptide-specific radio-immunoassays with antibodies against near-C-terminal and near-N-terminal portions of ANF prohormone. This suggests local ANF generation in neural structures. In spontaneously hypertensive rats (SHR) we found an elevated ANF-C content in all tissues along the pathway of increased efferent sympathetic outflow, which is present in this animal model. The ANF-N was augmented in SHR only in the hypothalamus. This indicates an overall increase of neural ANF in SHR. The reported neuroinhibitory function of increased neural ANF, however, was attenuated by a decrease in the number of some brain and peripheral ganglionic ANF binding sites in SHR. It remains to be determined whether the increased neural ANF in SHR is a primary phenomenon or a compensatory increase induced by high blood pressure. PMID- 2977164 TI - Effect of atrial natriuretic factor on arterial baroreceptor control of heart rate and blood pressure in conscious rats. AB - The effects of atrial natriuretic factor (ANF) on arterial baroreceptor control of heart rate and of blood pressure were examined in conscious normotensive rats chronically instrumented with arterial and venous catheters, by measuring (1) the pulse interval responses to four intravenous boluses of phenylephrine and four intravenous boluses of nitroprusside, reflex sensitivity being calculated as the slope of the linear relationship between pulse interval and mean arterial pressure (nine rats); (2) the pressor response to right common carotid occlusion (balloon-in-cuff occluder) in eight rats with aortic and left carotid baroreceptor denervation. The study was performed before and during a non hypotensive infusion of ANF. Atrial natriuretic factor increased the bradycardic responses to phenylephrine by 90% but reduced the tachycardic response to nitroprusside by 67% (P less than 0.01 for both) and left the pressor response to carotid occlusion unaffected (-7%, NS). It is concluded that ANF modulates the arterial baroreceptor reflex in a complex fashion, with opposite responses to arterial baroreceptor stimulation and deactivation, and different responses for the cardiac and vascular component of the reflex. PMID- 2977165 TI - Effect of changes in sodium balance on renin, angiotensinogen and atrial natriuretic factor messenger RNA levels in rats. AB - The expression of messenger (m)RNA for renin, angiotensinogen and atrial natriuretic factor (ANF) was investigated in rats on different sodium intakes. Messenger RNA was measured by a radiodensitometric hybridization assay. In the high-sodium state, renal renin mRNA decreased, but it increased in the low-sodium state. A further increase in renin mRNA was seen in the low-sodium state after captopril administration. Angiotensinogen mRNA levels in the liver, kidney and brain were altered by varying sodium intake. In the high-sodium state angiotensinogen mRNA decreased, but in the low-sodium state it increased. After treatment with captopril, angiotensinogen mRNA levels decreased in the liver and kidney. Angiotensinogen mRNA showed tissue specificity for expression, especially in the brain. Atrial ANF mRNA levels changed slightly with different levels of sodium intake. PMID- 2977166 TI - Nicardipine and atrial natriuretic factor increase whole body vascular permeability in rats. AB - Using binephrectomized anaesthetized rats, we explored the possibility that calcium antagonists may alter the partition of extracellular fluid between plasma and the interstitium, as suggested for atrial natriuretic factor (ANF). The effects of intravenous infusion of synthetic ANF (103-126 ANP; Wy 47.663) and the dihydropyridine derivative nicardipine were assessed by measuring changes in haematocrit and plasma proteins. After a 40-min infusion of ANF or nicardipine, haematocrit increased significantly (9% and 5.4%, respectively). The calculated loss of plasma volume was 15% after administration of ANF and 9.1% after nicardipine compared with 3.9% in rats receiving vehicle only. Plasma proteins increased by only 3.9% (after ANF) and 3.7% (after nicardipine), less than expected for a plasma volume contraction without protein extravasation. Atrial natriuretic factor and nicardipine induced a similarly slight change in mean arterial pressure. These results suggest that nicardipine and ANF both reduce plasma volume by an extrarenal mechanism; part of the fluid shift might be facilitated by an increased vascular permeability to proteins. PMID- 2977167 TI - Peptides derived from atrial natriuretic polypeptide precursor in human and monkey brains. AB - Using radio-immunoassays for the C-terminal sequence of alpha-atrial natriuretic polypeptide [atrial natriuretic factor (ANF)-(99-126)], alpha-ANP (17-28) [ANF (115-126)], which corresponds to gamma-human ANP [115-126; gamma-hANP-(115-126)] and for the N-terminal sequence of gamma-hANP [human ANF-(1-126)], gamma-hANP-(1 25) [human ANF-(1-25)], we detected the parallel distribution of alpha-hANP-like immunoreactivity and gamma-hANP-(1-25)-like immunoreactivity in the human and monkey brains, with the highest concentrations in the midbrain and the pons. High performance gel permeation chromatography coupled with the two radio-immunoassays and also with another radio-immunoassay using a monoclonal antibody against the N terminal sequence of alpha-ANP revealed that gamma-ANP is synthesized in the brain and cleaved into N-terminally deleted form(s) of alpha-ANP and 10-K N terminal fragment(s) of gamma-ANP, which coexist within the neuron. These results suggest that the post-translational processing of gamma-ANP in the brain is different from that in the heart. PMID- 2977168 TI - Increase in plasma atrial natriuretic factor and right atrial area during endogenous and exogenous volume loading in healthy volunteers: effect on plasma renin activity, aldosterone and antidiuretic hormone. AB - The mechanisms of atrial natriuretic factor (ANF) release and the effects of ANF on plasma renin activity (PRA), plasma aldosterone and plasma antidiuretic hormone were investigated in two groups of normal subjects. One group was studied before and after endogenous volume loading produced by 30 min of head-down tilt. The other group was studied before and after an exogenous volume load in the form of 1 litre of isotonic saline infused over 30 min. Measurements made included right atrial area by two-dimensional echocardiography, PRA, plasma aldosterone, plasma antidiuretic hormone and plasma levels of ANF. There was a significant correlation between the increase in atrial area and plasma ANF concentrations after endogenous and exogenous volume loading. Levels of PRA and plasma aldosterone were suppressed significantly after exogenous volume loading but not after endogenous volume loading. The concentration of plasma antidiuretic hormone was not affected by either procedure. Short-term changes in right atrial pressure appear to regulate the plasma levels of ANF in normal people. PMID- 2977169 TI - Effects of calcium and ouabain on the release of atrial natriuretic factor. AB - In order to examine the role of calcium in the secretory process of atrial natriuretic factor (ANF), we studied the effects of hypercalcaemia and ouabain on the plasma concentration of immunoreactive ANF, and the effect of calcium on immunoreactive ANF release from isolated rat atria. Anaesthetized dogs were treated with CaCl2 infusion, ouabain or phenylephrine injection. With CaCl2 infusion, serum calcium and plasma immunoreactive ANF respectively increased to three and four times their basal levels. Ouabain increased plasma immunoreactive ANF to two and a half times the initial level. Neither CaCl2 nor ouabain produced any effect on right atrial pressure and heart rate, but they both significantly increased arterial pressure. Phenylephrine caused a greater increase in arterial pressure than both CaCl2 and ouabain. However, there was no significant increase in plasma immunoreactive ANF. Moreover, calcium stimulated the release of immunoreactive ANF from isolated rat atria. These results suggest that the calcium may play a key role in the secretory process of ANF. PMID- 2977170 TI - Preparation of monoclonal antibodies against atrial natriuretic polypeptide precursor and application to highly sensitive sandwich enzyme immunoassay. AB - We produced three monoclonal antibodies against different epitopes of the precursor form of atrial natriuretic polypeptide (ANP), gamma-human ANP (gamma hANP) [human atrial natriuretic factor (ANF)-(1-126)]. Monoclonal antibodies were prepared by fusion of mouse myeloma cells X63-Ag8.653 with spleen cells from BALB/c mice immunized with synthetic alpha-hANP or gamma-hANP-(1-25) [human ANF (1-25)] conjugated to thyroglobulin. Of three monoclonal antibodies obtained, two were directed against the alpha-ANP sequence: one antibody (KY-ANP-I) recognizes the N-terminal half of the ring structure, while the other (KY-ANP-II) recognizes the N-terminal sequence of alpha-hANP. The third antibody (KY-ANP-III) was against gamma-hANP-(1-25). With the aid of rabbit polyclonal anti-alpha-ANP-(17 28), we developed sandwich enzyme immunoassays. An enzyme immunoassay for alpha hANP using KY-ANP-I and polyclonal Fab' was very sensitive (0.01 fmol/tube) and we could detect 0.6 pg/ml of plasma alpha-hANP without extraction. Thus, monoclonal antibodies against gamma-ANP are useful tools for investigating the significance of ANP and related peptides derived from gamma-ANP. PMID- 2977171 TI - A new Australian kindred with the syndrome of hypertension and hyperkalaemia has dysregulation of atrial natriuretic factor. AB - A family with the syndrome of hypertension and hyperkalaemia affecting six members in two generations is reported from Australia, where the first two sporadic cases were described. All family members had hyperkalaemia, hyperchloraemia and normal creatinine clearance. Only one affected adult and no affected children were hypertensive, possibly because of habitual low-salt diets. Plasma potassium fell significantly during fludrocortisone acetate administration, and urine potassium increased during saline infusion, consistent with renal tubular responsiveness to mineralocorticoid. Low plasma renin activity and pressor hyper-responsiveness to angiotensin II suggested sodium volume overload, but atrial natriuretic factor (ANF) was normal or only slightly elevated when compared with clearly elevated levels in primary aldosteronism. Plasma ANF was unresponsive to the usually reliable stimulus of angiotensin infusion in the two brothers affected and to saline infusion in one of them. These findings are consistent with a renal tubular avidity for sodium, leading to volume expansion, suppression of renin, and, depending on dietary sodium intake, hypertension. A role for dysregulation of ANF in the pathophysiology is possible. PMID- 2977172 TI - Cosecretion of peptides derived from gamma-human atrial natriuretic polypeptide in normal volunteers and patients with essential hypertension and adrenal disorders. AB - Using two radio-immunoassays for N-terminal and C-terminal fragments of human atrial natriuretic polypeptide (ANP) precursor, gamma-hANP [human atrial natriuretic factor-(1-126)], that is gamma-hANP(1-25) [human atrial natriuretic factor-(1-25)] and alpha-hANP [human atrial natriuretic factor-(99-126)], we studied the secretion of gamma-hANP-derived peptides into circulation from the heart in normal subjects and patients with essential hypertension and adrenal disorders. Volume expansion with 2 litres physiological saline increased plasma gamma-hANP(1-25)-like immunoreactivity concomitantly with plasma alpha-hANP-like immunoreactivity in normal subjects. Infusion of angiotensin II (20 ng/kg per min) or noradrenaline (200 ng/kg per min) also caused a parallel increase in plasma gamma-hANP(1-25)-like and alpha-hANP-like immunoreactivity. Plasma gamma hANP(1-25)-like immunoreactivity levels were changed together with alpha-hANP like immunoreactivity in patients with essential hypertension and adrenal disorders. These results indicate that gamma-hANP-derived peptides, alpha-hANP and the 10-k N-terminal fragment of gamma-hANP (N-peptide) are cosecreted from the heart and that the simultaneous measurement of N-peptide and alpha-hANP serves as an indicator of the cardiac endocrine function. The significance of N peptide as a hormone must await further clarification. PMID- 2977173 TI - Atrial wall stress rather than pressure per se might be responsible for the increased secretion of atrial natriuretic factor after heart transplantation. AB - Plasma atrial natriuretic factor (ANF) concentrations were measured, and relationships to intracardiac pressures and atrial dimensions were assessed in a series of 17 heart transplant recipients undergoing cardiac catheterization during their annual evaluation. Despite excellent cardiac function (normal filling pressures, adequate cardiac output, ejection fraction above 60%), plasma levels of ANF were elevated. Step-up levels across the heart were consistent with increased cardiac secretion, whereas both the metabolic clearance rate and plasma half-life were normal. Correlations between plasma concentrations of ANF at different cardiac sites and atrial filling pressures were low. However, right and left atrial dimensions in our patients were greatly enlarged. These findings suggest that increased atrial size (by virtue of the atrial anastomoses) and augmented wall stress (law of Laplace) rather than pressure per se are responsible for the increased ANF production after heart transplantation. PMID- 2977174 TI - The vasodilator effect of human atrial natriuretic factor (99-126) on human omental arteries. AB - The effect of synthetic human atrial natriuretic factor [ANF-(99-126)] on human omental arteries was investigated. The compound produced concentration-dependent relaxation, 10, 50 and 90% of maximum effect being observed at about 1, 10 and 100 nmol/l, respectively. These concentrations are considerably higher than those measured by radio-immunoassay in human plasma, even under extreme conditions (0.001-0.3 nmol/l). Therefore, ANF can dilate human resistance-size arteries, but whether it does so under physiological conditions has not been established. PMID- 2977175 TI - Dopaminergic control of aldosterone secretion is not mediated by atrial natriuretic factor in patients with essential hypertension. AB - Both dopamine and atrial natriuretic factor (ANF) are known to suppress aldosterone secretion. Since it is possible that dopaminergic mechanisms facilitate ANF release, we investigated the relationship between these two inhibitory systems by comparing the increases in aldosterone induced by metoclopramide, a dopaminergic antagonist, with decreases in ANF. Aldosterone, ANF, prolactin, plasma renin activity, cortisol and potassium were measured before and after the intravenous injection of 10 mg metoclopramide, blood samples being collected at 15-min intervals up to 2 h after the injection. These studies were performed in patients with essential hypertension who were maintained on a constant sodium intake (100 mmol/day), before and after 5 days of treatment with ibopamine, an orally active dopamine analogue. Before ibopamine metoclopramide induced the expected, marked increases in aldosterone and in prolactin, but only minimal, non-significant decreases in ANF. All other humoral parameters, as well as blood pressure and heart rate, were unaffected by metoclopramide. After ibopamine treatment, which caused a transient natriuretic effect, the responses of aldosterone and of ANF to metoclopramide were similar to those observed in control studies, whereas that of prolactin was enhanced. Thus, it appears that the suppressive effect exerted by the dopaminergic tone on aldosterone secretion is independent of ANF both before and after dopaminergic stimulation. PMID- 2977176 TI - Atrial natriuretic factor protects the isolated working ischaemic rat heart against the action of angiotensin II. AB - The interaction between atrial natriuretic factor [synthetic human ANF-(103-126)] and angiotensin II (Ang II) and its influence on reperfusion arrhythmias, cardiodynamics, enzyme loss and metabolic changes were investigated in isolated ischaemic working rat hearts. Acute regional myocardial ischaemia was induced by coronary artery occlusion which was associated with ventricular fibrillation. Perfusion with 1 X 10(-9) mol/l Ang II markedly aggravated these arrhythmias. Perfusion with 1 X 10(-7) mol/l ANF, in contrast, gave protection against ventricular fibrillation and prevented Ang II-induced aggravation of ventricular fibrillation. Atrial natriuretic factor improved cardiodynamics, in particular, during reperfusion, whereas Ang II impaired cardiodynamics and increased the release of creatine kinase and lactate dehydrogenase. These adverse effects of Ang II were absent when ANF was simultaneously perfused. Compared with control hearts, myocardial tissue levels of glycogen, ATP and creatine phosphate were increased in hearts perfused with either ANF or ANF plus Ang II, whereas lactate levels decreased. Perfusion with Ang II alone led to deterioration in these metabolic parameters. These results in isolated working rat hearts suggest that ANF protects against the consequences of ischaemia and reperfusion and that functional antagonism between ANF and Ang II may contribute to this. PMID- 2977177 TI - Dopaminergic-opioidergic interaction is reflected by changes in pituitary hormone secretion in patients with essential hypertension. AB - In a randomized, double-blind crossover study 13 untreated patients with mild essential hypertension were exposed to submaximal bicycle exercise. Sixty minutes before ergometry 10 mg metoclopramide or placebo, and 10 min before exercise 0.4 mg naloxone or placebo, were given intravenously. Plasma adrenocorticotrophic hormone, beta-endorphin and cortisol levels increased significantly after ergometry, whether performed after placebo, naloxone, metoclopramide or metoclopramide + naloxone treatment. However, only naloxone administration potentiated plasma adrenocorticotrophic hormone, beta-endorphin and cortisol responses to workload. Plasma levels of adrenocorticotrophic hormone, beta endorphin and cortisol were 45 +/- 14 pg/ml, 6.2 +/- 1.2 pmol/l and 141 +/- ng/ml, respectively, after ergometry, when performed after placebo, but these values were increased to 61 +/- 10 pg/ml, 11.4 +/- 2.8 pmol/l and 207 +/- 22 ng/ml, respectively, after naloxone treatment. This naloxone-induced potentiation of hormonal release was blocked by metoclopramide pretreatment, suggesting a close interaction between dopaminergic and opioidergic mechanisms, regulating hormonal responses to physical exercise. PMID- 2977178 TI - Platelet vasopressin receptor in essential hypertension. AB - The specific vasopressin receptor of V1 vascular subtype, which mediates platelet aggregation, has been found on human platelets. We investigated the binding characteristics using tritiated arginine vasopressin [3H]-AVP and platelet aggregation with AVP turbidometrically in normal subjects, patients with WHO class II essential hypertension and patients with malignant-phase hypertension. In essential hypertensives Bmax was significantly higher than that in normal subjects, but there were no differences in affinity and the maximal percentage aggregation between them. In malignant-phase hypertensives Bmax and maximal percentage aggregation were significantly lower than those in normals and essential hypertensives, although there was no difference in the affinity between them. With radio-immunoassay, the mean platelet-free plasma AVP level was significantly higher in malignant-phase hypertensives than those in normals and essential hypertensives, whereas there was no difference in mean platelet AVP levels between them. In essential hypertensives Bmax and maximal percentage aggregation did not change, but in malignant-phase hypertensives Bmax increased significantly and maximal percentage aggregation tended to normalize after treatment. PMID- 2977179 TI - Vasopressin receptors in rat brain and kidney: studies using a radio-iodinated V1 receptor antagonist. AB - Arginine8-vasopressin (AVP) acts via V1 receptors (blood vessels, liver and brain) and V2 receptors (renal collecting duct). To study brain and kidney V1 receptors selectively, a specific V1 receptor antagonist [d(CH2)5,Sar7]AVP was radio-iodinated and purified by high performance liquid chromatography. Iodine 125[d(CH2)5,Sar7]AVP bound to single classes of rat liver and kidney V1 receptors with high affinity (liver: Kd = 3.0 +/- 0.9 mol/l and Bmax = 530 +/- 10 fmol/mg protein; kidney: Kd = 0.5 +/- 0.9 nmol/l and Bmax = 11 +/- 8 fmol/mg protein) in a time-dependent and saturable manner. Displacement of the radioligand from liver and renal medulla membranes and sections of the brain and kidneys by unlabelled AVP analogues was consistent with that expected for binding to V1 receptors. In vitro autoradiography of rat brain revealed areas of specific receptor binding in many regions, including regions involved in central cardiovascular regulation, such as the nucleus of the solitary tract and area postrema, as well as choroid plexus and large blood vessels. Binding was observed in several regions not previously observed to contain AVP receptors. In the kidney [3H]AVP bound to the inner and outer medulla, probably to vascular V1 and collecting duct V2 receptors. In contrast, [125I][d(CH2)5,Sar7]AVP binding was only in the inner medulla, possibly to vasa recta. These findings support a functional role for V1 receptors in the brain and kidney. PMID- 2977180 TI - Vasodilation induced by a specific vasopressin V2 agonist in dogs. AB - Vasopressin V2 agonists increase cardiac output and lower total peripheral resistance within minutes when infused into conscious dogs. The purpose of this study was to determine the mechanisms of these haemodynamic effects. Two groups of dogs were studied. In the first group, six dogs were instrumented for the measurement of mean arterial pressure and cardiac output (electromagnetic flowmeter) at least 1 week before the experiment. They were then anaesthetized with pentobarbital and the central nervous system was destroyed. Mean arterial pressure was maintained with an infusion of noradrenaline. After it had remained stable for 30 min the dogs received an intravenous infusion of 4-valine-8-D arginine vasopressin (VDAVP), a specific V2 agonist, 10 ng/kg per min. Mean +/- s.e.m. arterial pressure fell from 97.6 +/- 1.8 to 69.1 +/- 2.5 mmHg, but cardiac output did not change significantly. Total peripheral resistance fell from 1322.5 +/- 151 to 859.7 +/- 84.9 units. Thus, VDAVP exerts haemodynamic effects in the absence of the central nervous system and appears to act primarily as a vasodilator. A second group of six anaesthetized dogs was studied in which femoral blood flow was measured with an electromagnetic flow probe during intravenous and intrafemoral arterial infusion of VDAVP, 10 ng/kg per min. Blood flow increased by 27.9 +/- 5.4 ml/min from a control value of 65.1 +/- 12.6 ml/min with intra-arterial infusion of VDAVP, but did not change significantly with intravenous infusion. Mean arterial pressure remained unchanged under both conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977181 TI - Prediction of the progression of cardiac hypertrophy in middle-aged mild hypertensives. AB - To investigate the predictive value of exercise tests and diastolic function measurements for the progression of left ventricular hypertrophy, symptom-limited treadmill stress testing and echocardiography were performed before and after a follow-up period of 3.5 years in 47 mild hypertensive men aged 42 +/- 2 years. The men were classified into three groups by the progression of the left ventricular mass index (%LVMI) during the observation, i.e. (LVMI after follow up) - (LVMI before follow-up)/(LVMI before follow-up). The high-progression group (n = 13) had a %LVMI exceeding mean +/- 2/3s.d. of all subjects; the low progression group (n = 21) had a %LVMI within mean +/- 2/3s.d. and the non progression group (n = 13) had a %LVMI less than mean -2/3s.d. At the beginning of the observation, age, blood pressure at rest, LVMI, ejection fraction, mean velocity of circumferential fibre shortening, peak shortening rate and systolic time intervals (ET/PEP, ratio of ejection time to pre-ejection period) were similar among the three groups. However, the high-progression group showed a higher systolic pressure at peak exercise, a lower peak filling rate and a longer time to peak filling rate (TPFR) as corrected by the R-R interval of the ECG. These data suggest that systolic pressure at peak exercise and echocardiographically assessed diastolic function are useful in predicting the progression of cardiac involvement in mild hypertension. PMID- 2977182 TI - Long-term antihypertensive treatment may induce normalization of left ventricular mass before complete regression of vascular structural changes: consequences for cardiac function at rest and during stress. AB - In 14 essential hypertensive patients, aged 26-59 years, blood pressure, left ventricular mass index (LVMI), systolic function (M-mode echo, two-dimensionally guided), post-ischaemic 'maximal' forearm blood flow (strain gauge venous occlusion plethysmography), plasma renin activity, plasma catecholamines and aldosterone were measured before and after 6 and 12 months of treatment (eight patients were given captopril, 100 mg/day, + hydrochlorothiazide 25 mg/day in five patients, and six patients were given nitrendipine, 20 mg/day, + atenolol 50 mg/day in four patients). Minimal vascular resistance (mean blood pressure/peak forearm blood flow) was taken as an index of arterial structural changes. After 6 months of treatment significant reductions in blood pressure (P less than 0.001), LVMI (P less than 0.001) and minimal vascular resistance were observed. After 12 months of treatment blood pressure, LVMI and minimal vascular resistance were further reduced. The LVMI was normalized in nine cases and the minimal vascular resistance in two cases only. Aldosterone and plasma catecholamines did not change, whereas plasma renin activity was increased during captopril only. Before and during treatment the left-ventricular shortening fraction in relation to end systolic stress in each patient at rest, and at peak of handgrip and cold pressor tests, fell within the 95% confidence limits of correlation obtained in normals. Thus, in essential hypertensives long-term treatment can induce normalization of LVMI before complete regression of arterial structural changes in the forearm. Left ventricular systolic function is preserved after normalization of LVMI, both at rest and during stress. PMID- 2977183 TI - Left ventricular systolic function in relation to withdrawal of different pharmacological treatments in hypertensives with left ventricular hypertrophy. AB - We evaluated the left ventricular mass index (LVMI) and the functional response to cold pressor and handgrip tests in 74 untreated essential hypertensive patients and 26 age and sex-matched normals. The same measurements were repeated in 22 essential hypertensives after 6 and 12 months of treatment (captopril or nitrendipine, plus diuretic or beta-blocker in a few cases) and in 21 essential hypertensives after withdrawal of treatment, a reduction in the LVMI and a further increase in blood pressure. Left ventricular systolic function was evaluated by the relationship between left ventricular end-systolic stress and fractional shortening. Highly significant negative correlations, with similar slopes and intercepts, were found between end-systolic stress and fractional shortening under basal conditions, after regression of left ventricular hypertrophy and after withdrawal of treatment, both at rest and at the peak of stress tests. An examination of each point of the relation between end-systolic stress and fractional shortening showed that very few points were beyond the 95% prediction limits of the correlation obtained in normal volunteers. These results indicate that left ventricular systolic function is normal in most untreated essential hypertensives, and is usually well maintained after regression of left ventricular hypertrophy during long-term treatment as well as after withdrawal of treatment, both at rest and during an acutely induced afterload increase. PMID- 2977184 TI - Photoproducts formed from photofrin II in cells. AB - Fluorescence and absorption spectra of light-exposed cells containing the tumour localizing porphyrin preparation Photofrin II (PII) have been studied. Light exposure results in spectral changes that may be due to a photoinduced modification of the porphyrins without breakage of the porphyrin macrocycle and/or to a photoinduced displacement of the porphyrins in the cells. Photochemical reaction involving breakage of the porphyrin macrocycle also occur as can be seen from the loss of absorbance within the Soret band region during light exposure. Singlet oxygen may be involved in the photodegradation of PII in cells since the process is slowed down on bubbling N2 through the samples and is slightly faster in suspensions in Dulbecco's phosphate buffered saline (PBS) made of D2O compared with suspensions in PBS made of H2O. During light exposure a fluorescent product is formed in the cells with fluorescence excitation and emission characteristics similar to those of the "age pigment" lipofuscin (lambda exc = 350 nm, lambda em = 440 nm). PMID- 2977185 TI - Photoreactivation in phr mutants of Escherichia coli K-12: cloning from the gal att lambda interval increases the photoreactivable response in phrA strains. AB - In this report we have cloned restriction fragments from the gal-att lambda region obtained from a purified preparation of lambda dgal transducing 'phage DNA, and demonstrate the appearance of a photoreactivable response in a photoreactivation-deficient phrA phrB strain. We also show that when this plasmid is transduced into a delta phrA strain there is an increase in the photoreactivable response after a single high intensity light flash and after continuous illumination. These data have been discussed in relation to the hypothesis of the presence of multiple photolyase molecules in Escherichia coli. PMID- 2977186 TI - Blue light induces circadian rhythms in the bd mutant of Neurospora: double mutants bd,wc-1 and bd,wc-2 are blind. AB - This paper describes a new blue light effect for Neurospora crassa, the photoinduction of circadian rhythms in the bd mutant. The wc-1 and wc-2 genes are necessary for this effect. PMID- 2977187 TI - Is human atrial natriuretic peptide in fetal blood useful as a parameter to detect the decompensate state of the fetal heart? AB - In nonimmunologic hydrops fetalis associated with heart anomaly, plasma human atrial natriuretic peptide (hANP) values in umbilical venous blood were significantly higher than those in normal infants. These findings indicate that significant release of hANP from atria is induced by the decompensatory state of the fetal heart. PMID- 2977188 TI - Immunogenetic and racial determinants of gold toxicity in rheumatoid arthritis. AB - Typing for antigens HLA-A,B,C and DR was performed on 165 rheumatoid arthritis patients (14 black, 151 white) who had received gold therapy to determine the relationship between HLA antigens and gold dermatitis, stomatitis, thrombocytopenia, and proteinuria. Dermatitis and stomatitis occurred in both black and white patients. Thrombocytopenia and proteinuria occurred only among the white patients studied. The absence of thrombocytopenia and proteinuria among the black patients was not statistically significant. Antigen HLA-DR7 was uncommon among black and white subjects with dermatitis (0 of 6 blacks, 4 of 48 whites), but this decrease in frequency was not statistically significant. Antigen HLA-DR3 was an important risk factor for thrombocytopenia (relative risk = 11.8, P = .0043) and proteinuria (RR = 5.8, P = .032). These results are consistent with previous studies of HLA-DR3 and gold toxicity. The only black patient with stomatitis possessed the A1B8DR3 phenotype. Future studies should examine whether the same HLA antigen confers risk of different gold toxicities in different racial groups, and whether there are HLA antigens that provide a protective effect. PMID- 2977190 TI - Aggressive management of acute myocardial infarction. AB - Acute transmural myocardial infarction is usually caused by a coronary thrombus along with fixed coronary artery stenosis. Myocardial necrosis can be interrupted by the prompt use of pharmacologic and mechanical thrombolysis. Intravenous streptokinase and urokinase have been associated with approximately a 45 percent recanalization rate while the newer agent, recombinant human tissue-type plasminogen activator (rt-PA), has an average lysis rate of 70 percent intravenously. Intracoronary streptokinase and urokinase have a similar 75 percent lysis rate, but with additional costs and morbidity. Percutaneous transluminal angioplasty (PTCA) is often indicated to correct an underlying stenosis, the time of which depends on the experience and expertise of the PTCA team. PMID- 2977189 TI - Carbon dioxide laser laparoscopy in treatment of infertility and disorders associated with pelvic pain. AB - Seventy-five patients, from December 1984 to December 1985, received carbon dioxide (CO(2)) laser laparoscopy for infertility and pelvic pain. The chief complaint of 55 patients was pelvic pain, and for 20 patients, either primary or secondary infertility. The most common findings were endometriosis (84 percent) and pelvic adhesions (35 percent).This paper gives the incidence of multiple diagnostic findings and the use of CO(2) laser laparoscopy. The results indicate that with the availability of the CO(2) laser laparoscope a significant number of patients can be treated for endometriosis, pelvic adhesions, salpingitis, and other disorders, preventing the need for future surgical procedures or medical therapy. PMID- 2977191 TI - Left ventricular end-systolic wall stress--dimension relationship in unanesthetized dogs with perinephritic hypertension. AB - To study myocardial contractility in hypertensive hearts with normal wall motion, we examined left ventricular end-systolic wall stress-dimension relationships (ESWDR) during a baseline period (CS: control stage) and in the eighth week after induction of systemic hypertension by Page's method (HS: hypertensive stage) in unanesthetized dogs. The mean aortic blood pressure increased from 94 +/- 11 to 142 +/- 26 mmHg (p less than 0.01). The end-diastolic left ventricular posterior wall thickness increased significantly during the HS (9.4 +/- 1.3 vs 7.3 +/- 1.3 mm; HS vs CS), and its dimension was significantly (p less than 0.05) smaller than it was during the CS (37.0 +/- 4.2 vs 39.9 +/- 4.6 mm; HS vs CS). There were no significant differences between the 2 stages in left ventricular fractional shortening (31.9 +/- 5.0 vs 32.6 +/- 2.8; HS vs CS), in end-systolic meridional left ventricular wall stress (75.3 +/- 10.8 vs 68.3 +/- 15.6 10(3) dynes/cm2; HS vs CS), or in the ESWDR slopes (98.6 +/- 17.7 vs 94.0 +/- 19.7; HS vs CS). The ESWDR dimension intercepts significantly decreased from 2.0 +/- 0.3 to 1.8 +/- 0.3 cm during the HS; that is, the relationship shifted to the left with no significant change in the slope. At autopsy, the ratio of left ventricular weight to body weight of the hypertensive dogs was significantly (p less than 0.01) greater than that of sham-operated control dogs (6.0 +/- 0.9 vs 4.3 +/- 0.5 g/kg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977192 TI - Orthostatic hypertension due to coexistence of renal fibromuscular dysplasia and nephroptosis. AB - A 42-year-old woman presented with orthostatic hypertension. Increased plasma renin activity was noted and blood pressure rose gradually with standing. Selective renal arteriography indicated narrowing of the distal portion of the right renal artery and poststenotic dilatation and signs of arterial stenosis due to fibromuscular dysplasia. Greater arterial narrowing resulted from tortion due to nephroptosis brought about by excessive renin secretion. Thus, both renal arterial stenosis and nephroptosis were considered responsible for the present orthostatic hypertension. Percutaneous transluminal renal angioplasty was found very effective for normalizing standing blood pressure and renal blood flow. PMID- 2977193 TI - Physiological factors of atrial natriuretic polypeptide release and its neural regulation in conscious dogs. AB - We have examined physiological factors in atrial natriuretic polypeptide (ANP) release and whether or not the cardiac nerves control release of ANP. Two possible factors were tested, an increase in plasma sodium level (PNa) and an increase in atrial pressure. Injection of 1.0 or 2.0 mEq/kg of sodium ions elevated PNa by 5.3 +/- 0.3 or 7.3 +/- 0.4 mEq/L, respectively, but plasma ANP level (PANP) did not change. Infusion of 18 ml/kg of 3% Dextran-40 over 5 min increased mean left atrial pressure (MLAP) by 7.6 +/- 0.9 mmHg. PANP increased from 206 +/- 17 pg/ml to 260 +/- 25 pg/ml, which was not significant. PANP, corrected for hemodilution, significantly increased to 348 +/- 34 pg/ml. These results suggest that PNa increase does not promote ANP release, but that an atrial pressure increase does. This transient volume load did not induce full response of the ANP releasing system. A prolonged volume load for 45 min increased corrected PANP to 435 +/- 73 pg/ml. A close linear correlation was found between the increases in MLAP and PANP. These facts indicate that prolonged volume expansion is necessary to induce full response of the ANP releasing system. Complete cardiac denervation did not affect the tonic level of plasma ANP, volume expansion-induced increase in PANP, or the sensitivity of the ANP releasing system. Thus we conclude that the cardiac nerves do not control ANP release caused by volume expansion. PMID- 2977194 TI - Role of atrial natriuretic peptide in experimental renal failure. PMID- 2977195 TI - Effects of chronically administered atrial natriuretic factor in aldosterone infused hypertensive rats. AB - To assess the pathophysiological role of atrial natriuretic factors in mineralocorticoid hypertension, we studied the effects of chronic infusion of synthetic atrial natriuretic factor on blood pressure and sodium-water excretion in rats with aldosterone salt-induced hypertension. Administration of synthetic atrial natriuretic factor (150 micrograms/kg/day) to rats made hypertensive by 7 day infusion of aldosterone (100 micrograms/kg/day) and sodium loading with 1% NaCl as drinking water returned the blood pressure to control levels, and the antihypertensive effect was not associated with any changes in urine volume and urinary sodium excretion. These results indicate that atrial natriuretic factors may be involved in the regulation of blood pressure in mineralocorticoid hypertension, independent of the renal effects of these substances. PMID- 2977196 TI - Factors influencing vascular and natriuretic responses to ANP. AB - To explore factors that modulate the magnitude of vasorelaxation and natriuretic/diuretic responses to exogenous ANP, clearance studies were performed in unilateral hydronephrotic rats with renal vasoconstriction complications. An increase in renal plasma flow associated with a decrease in calculated renal vascular resistance in response to ANP (2 micrograms.min-1.kg-1, i.v.) was obtained only in the hydronephrotic kidney. In contrast, the control kidney of the unilateral hydronephrotic rat had a decrease in glomerular filtration rate and an abolished natriuretic response following ANP administration. A prior renal denervation in the control kidney restored the natriuretic effect of ANP and abolished the lowering of glomerular filtration rate. These data suggest that the vasodilatory effect of ANP may depend on the vascular tone itself, and that renal responses to ANP are significantly modified by renal nerve activity. PMID- 2977197 TI - An integrated study employing histopathological, immunohistocytochemical and radioimmunoassay analyses of atrial natriuretic peptide in the right and left atria in patients with mitral valve disease. AB - To clarify the production mechanism of atrial natriuretic peptide (ANP) in right (RA) and left atria (LA) in mitral valve disease, histopathological and immunohistocytochemical analyses were performed and ANP levels were investigated by radioimmunoassay (RIA) in 28 patients. Atrial tissues were obtained during mitral valve replacement. ANP-like immunoreactivity of the myocytes applied by the avidin-biotin peroxidase complex method was observed around the nuclei of the atrial myocytes. Electronmicroscopically, immunoreactivity was observed in atrial specific granules. Light-microscopically determined intensity of the immunoreactivity was classified into 4 grades and the intensity in 100 myocytes was expressed by adding the scores of each myocyte. Mean right atrial pressure was positively correlated with the activity score in RA (r = 0.80). Pulmonary capillary wedge pressure was not correlated with the score in LA. The score in RA was significantly higher than that in LA. The ANP level in RA investigated by RIA was also higher than that in LA. Histopathological findings such as myocyte hypertrophy, degeneration and interstitial fibrosis were more severe in LA than in RA. In conclusion, longstanding atrial overloading, especially in LA, caused severe pathological damage, resulting in a smaller production of ANP. Much more ANP may be produced from RA in long-standing mitral valve disease. PMID- 2977198 TI - Physiological significance of atrial natriuretic peptides in essential hypertension. AB - To investigate the significance of atrial natriuretic peptides (ANP) in essential hypertension, plasma ANP concentrations in 43 essential hypertensives, 16 borderline hypertensives and 17 normotensive controls were measured. Furthermore, effects of high-sodium and low-sodium intakes on plasma ANP concentration were examined in "salt-sensitive" [SS] and "nonsalt-sensitive" [NSS] patients with essential hypertension. Plasma ANP concentration was significantly higher in hypertensives than in borderline hypertensives and in normotensive controls. No significant difference in plasma ANP concentration was observed between borderline hypertensives and normotensive controls. Plasma ANP concentration increased with the high-sodium diet in both the SS and NSS patients, but the mean increment was significantly greater in the SS than the NSS patients. Urinary excretion of sodium was lower in the SS patients taking the high-sodium diet than the corresponding value in the NSS patients. These findings suggest that an increased level of circulating ANP in hypertensive patients represents a compensatory mechanism to offset further elevation of blood pressure and sodium retention. PMID- 2977199 TI - Effect of exercise on circulating atrial natriuretic peptide and left ventricular ejection fraction in healthy persons and patients with coronary artery disease. AB - Radionuclide angiographic measurements of left ventricular ejection fraction were performed at rest and during exercise in 10 normal persons and 11 patients with coronary artery disease. Exercise was continued on a supine bicycle exercise table up to a symptom-limited maximum. Plasma levels of atrial natriuretic peptide (ANP) were also determined at rest and during exercise. Ejection fraction in the normal volunteers was 59 +/- 3% (mean +/- SEM) at rest and increased significantly (p less than 0.01) to 69 +/- 3% during exercise. Ejection fraction in the patients was 47 +/- 5% at rest and did not change significantly during exercise (51 +/- 7%). Plasma ANP in the normals rose significantly (p less than 0.01) from 62 +/- 16 pg/ml at rest to 454 +/- 94 pg/ml during exercise. Plasma ANP in the patients also rose significantly (p less than 0.01) from 231 +/- 102 pg/ml to 794 +/- 170 pg/ml. The response of plasma ANP to exercise was enhanced significantly (p less than 0.05) in the patients as compared with the normals in relation to ejection fraction by analysis of covariance. In both the normals and the patients, plasma ANP was inversely and significantly correlated with ejection fraction during exercise (r = -0.46, p less than 0.05, n = 21), however, not at rest. Because it has been reported that plasma ANP is correlated positively with pulmonary artery wedge pressure, the estimation of plasma ANP during an exercise stress test might be used for the evaluation of cardiac reserve in coronary artery disease. PMID- 2977200 TI - [Applications of the shape memory alloy in clinical medicine]. PMID- 2977201 TI - [Hemodynamics in obesity]. PMID- 2977203 TI - [Functions and structure of complement inactivators in plasma. 4). Factor H]. PMID- 2977202 TI - [Functions and structure of complement inactivators in plasma. 2). I factor]. PMID- 2977204 TI - [Functions and structure of complement inactivators in membrane proteins. 1). C3b/C4b receptor]. PMID- 2977205 TI - [Progress in the study of complement receptors. Structure and physiologic functions of complement receptors]. PMID- 2977206 TI - [Clinical study of congenital abnormality of complement system regulator factors]. PMID- 2977207 TI - [A bullous erythema multiforme type drug eruption induced by tiopronin showed pemphigus like eruption]. PMID- 2977209 TI - [A new laser Doppler velocimeter with two fibers and its application to measurements of stenotic coronary artery flow]. PMID- 2977208 TI - [Quantitative and functional study of peripheral lymphocytes in patients with alopecia areata]. PMID- 2977210 TI - [Circulating immune complexes in retinitis pigmentosa]. PMID- 2977211 TI - Bad backs--uncovering the real problem: the Alexander technique sheds new light on a sore situation. PMID- 2977212 TI - Assay of branched-chain alpha-keto acid dehydrogenase kinase in mitochondrial extracts and purified branched-chain alpha-keto acid dehydrogenase complexes. PMID- 2977213 TI - Significance of biogenic amines in functional disturbances resulting from brain injury. AB - Focal cortical freezing lesions in rats caused a widespread decrease in local cerebral glucose utilization (LCGU) in cortical areas of the lesioned hemisphere and this was interpreted as reflecting a depression of cortical activity (Pappius, 1981). Cortical serotonin (5-HT) metabolism was increased throughout the lesioned hemisphere (Pappius and Dadoun, 1987). In contrast, norepinephrine (NE) was decreased bilaterally, while levels of dopamine and its metabolites were not affected (Pappius and Dadoun, 1986). To determine if the changes in these neurotransmitters are of functional importance and mediate the observed changes in LCGU, the effects of inhibition of 5-HT synthesis with p-chlorophenylalanine (PCPA) and alpha 1-adrenergic blockage with prazosin (PZ) on cerebral metabolism and biogenic amine content in injured brain were studied. At doses of PCPA ineffective on LCGU (50 and 100 mg/kg) brain trauma still resulted in increased 5 HT metabolism. PCPA at doses which selectively ameliorated the depression of cortical LCGU in the lesioned hemisphere (200 and 300 mg/kg) completely prevented changes in 5-HT and 5-hydroxyindoleacetic acid seen following traumatization in untreated animals. These results provide evidence that decreased LCGU in lesioned brain is due to an activation of the serotonergic system. Prazosin (1 mg/kg) given 30 min before the lesion significantly increased cortical glucose utilization in the injured hemisphere and was even more effective when the treatment was continued for 3 days. Prazosin did not modify changes in cortical biogenic amines seen in untreated animals. The data are in agreement with a postulated inhibitory role of serotonin and norepinephrine in the cerebral cortex and implicate both neurotransmitters in functional alterations associated with injury. PMID- 2977214 TI - [Anesthesiologic considerations concerning the severely handicapped undergoing dental treatment under general anesthesia]. PMID- 2977215 TI - Beta-endorphin-like immunoreactivity increases in cerebrospinal fluid of acute head-injured patients. AB - beta-endorphin-like immunoreactivity (beta-ELI) was measured in cerebrospinal fluid (CSF) of 36 acute head-injured patients and 12 patients without head injury as controls. The mean level of beta-ELI in CSF of controls, mild cerebral contusions, and severe cerebral contusion patients were 51.9 +/- 5.6 pg/ml, 110.5 +/- 14.5 pg/ml, and 173.8 +/- 20.1 pg/ml respectively, with significant difference between them. The results also showed that beta-ELI may reflect the prognosis of acute head-injured patients. PMID- 2977217 TI - Safety: patient handling in the community. PMID- 2977216 TI - Erythroderma. PMID- 2977218 TI - Safe lifting and moving for nurse and patient. PMID- 2977219 TI - Rapid rise of serum testosterone following discontinuation of long term treatment of prostate carcinoma with an LHRH-agonist. PMID- 2977220 TI - A biometrical view on normal values of CD4 and CD8 lymphocyte counts in peripheral blood. AB - Statistical methods have been used to determine an optimal approach to the definition of the reference range for CD4 ("helper") and CD8 ("suppressor/cytotoxic") T cell numbers and the CD4/CD8 ratio in the peripheral blood. A graphical presentation of the absolute values for CD4 and CD8 for 85 healthy blood donors showed that a reference ellipse defined by fitting a gaussian distribution to logarithmically transformed data for absolute counts of CD4 and CD8 cells was superior to the fitting of an ellipse to untransformed data. Further analysis for another 147 subjects showed that the 95% tolerance prediction for the CD4/CD8 ratio in health could be stated with 95% confidence as 0.6 to 5.0. This approach allows clear definition of reference ranges for T cell tests in health and would also be applicable to results for patients with a disease such as HIV 1 infection in which a reference range for "well" patients exists and a change in the T cell ratio is of prognostic significance. PMID- 2977221 TI - Hairy cell leukemia cells are relatively NK-insensitive targets. AB - The hypothesis that interferon alpha (IFN alpha) has its beneficial effects in hairy cell leukemia by activating natural killer cells against hairy cells was examined. Leukemic cells from patients with hairy cell leukemia were tested for their susceptibility to lysis by fresh and IFN alpha activated peripheral blood mononuclear (PBMC) cells from normal donors. All hairy cells tested were relatively insensitive to cytolysis by PBMC and IFN alpha activated PBMC. The low levels of 51Cr release obtained with a few donors was due to lysis of leukemic cells, not residual normal cells, and was mediated by a natural killer cell (T cell receptor independent) mechanism. Chronic lymphatic leukemic cells before and after treatment with phorbol ester were also resistant to cytolysis. Hairy cells were not susceptible to lymphokine activated killer (LAK) cells but were sensitive to lysis by antibody and complement. The insensitivity to cell mediated cytolysis against hairy cells was shown by cold target inhibition to be a lack of target recognition by NK cells. PMID- 2977222 TI - [Hepatitis B: is the current vaccination policy the correct one everywhere?]. AB - In this study we have tried to examine retrospectively all the babies born to HBsAg + mothers in the past nine years, in order to evaluate which is the real incidence of perinatal transmission of HBsAg, and then if the actual politic to vaccinate only these babies is right and sufficient. The results demonstrate that the perinatal transmission is not a major problem in our country, so we suggest that for improvement of public health is more advised the vaccination of all infants: those born to HBsAg + mothers at the birth, the others in the third month of life. PMID- 2977223 TI - [Current immunologic and functional research on the skin in occupational contact dermatitis]. AB - Immunological and functional studies are performed on workers engaged in the production of plastics, artificial fibres and dyes, in view of clarifying some mechanisms in the evolution of the contact dermatitis and the related with it early changes. The study establishes increased values of IgA and IgG in workers in contact with irritating substances and increased values of IgG in workers in contact with sensitizing substances. In the latter is registered a decrease in the active and the total rosettes. The functional investigations, performed on 292 workers point out to no essential deviations in pH values in the limits of the physiological variability. The alkaline resistance of the skin is decreased in 51% of the examined, who are in contact with irritating substances and in 29.7% exposed mainly to sensitizing substances. PMID- 2977224 TI - Photodynamic effects of hematoporphyrin derivative on the uptake of rhodamine 123 by mitochondria of intact murine L929 fibroblasts and Chinese ovary K1 cells. PMID- 2977225 TI - Thymidine uptake, incorporation and DNA polymerase activity in murine bladder tumor cell, MBT-2, exposed to UV activated dihematoporphyrin ether. PMID- 2977226 TI - Photodynamic therapy induced ultrastructural alterations in microvasculature of the rat cremaster muscle. PMID- 2977227 TI - [Interleukin-1: a multifunctional molecule in inflammation and immune response- its structure and function in defence system]. PMID- 2977228 TI - [A stroll through the world of restorative dentistry]. PMID- 2977229 TI - The common language of Katz's index of ADL in six studies of aged and disabled patients. AB - Aged and disabled patients' need for coordinated services has increased the importance of a common language for their ability to perform the activities of daily living (ADL). This article aims at presenting Katz's Index of ADL and the use of it in six selected studies in order to show the availability for different purposes in divergent contexts. Katz's index is based on developmental principles and includes six common personal activities ordered into a cumulative scale. The use of the instrument provided both qualitative and quantitative information about disability among rather severely disabled patients in short-term and long term hospital care, in nursing homes, in day-care and in home-care. The level of disability among the less disabled persons, who were independent in all of the six activities, were not differentiated by the index. A supplement of instrumental activities based on the same developmental principles is required, before a common language also for less disabled people can be established. PMID- 2977230 TI - Acute renal failure, eosinophilia thrombocytopenia and exfoliative dermatitis associated with captopril therapy. PMID- 2977231 TI - [Incidence of diphtheria (data of the results of the bacteriologic examination of patients with tonsillitis)]. PMID- 2977232 TI - [Myxomatous degeneration in cardiovascular diseases]. PMID- 2977233 TI - [Hemodynamics in patients with myocarditis]. PMID- 2977234 TI - [Prognosis of the risk of early alcohol dependence among male adolescents]. PMID- 2977235 TI - [The role of 24-hour ECG monitoring in the diagnosis and evaluation of the treatment of unstable stenocardia]. PMID- 2977236 TI - [Nonenzymatic fibrinolysis of the blood and urine in urologic diseases]. PMID- 2977237 TI - [Integration of 2 handicapped children into a collective nursery]. PMID- 2977238 TI - Endoscopic observations of the residual cavity after liver hydatid disease surgery. AB - Endoscopic inspection of the residual cavity after liver hydatid disease surgery has the potential of determining the cause of delayed cavity obliteration by providing information about the inner surface condition. We therefore performed endoscopy whenever there was an unsatisfactory rate of reduction of the residual cavity size. Over the last 6 years, 72 patients have been operated upon for liver hydatid disease. Of these, 42 had wide-bore catheter drainage of the echinococcal cavity and 30 had primary surgical closure. In 17 of 42 patients, endoscopy was performed 3-14 months postoperatively, because the cavity showed no evidence of reduction in size. A flexible choledochoscope was passed through the transcutaneous drainage fistula. Silk sutures were found four times and residual parasitic elements were found 3 times. These were all removed. In 13 patients, the cavity was inspected within 6 months postoperatively; their inner cavity surfaces were covered with white soft areolar tissue. The other 4 patients' cavities were examined within 6-14 months; hard fibrous connective tissue was found. Two patients required reoperation, and in both cases the cavity collapsed within 3-12 months postoperatively. We conclude, that endoscopy of the residual cavity is a useful procedure for both recognition and removal of the cause of delay in the cavity obliteration and can lead to collapse of the cavity. PMID- 2977239 TI - Synthesis of new fluorescent spirolactone derivatives: determination of their affinities for aldosterone receptors. AB - Fluorescent spirolactone derivatives are obtained by coupling 3-carboxylic coumarins to a spirolactone bearing a 3-hydroxypropyl chain in the 7 alpha position. The two esters prepared by this method are highly fluorescent (emission 383 and 408 nm). PMID- 2977240 TI - [The structure and dynamics of ophthalmic pathology based on office visit data in the city of Kineshma]. PMID- 2977241 TI - [Laparoscopic organic rheography in the diagnosis of disorders of visceral blood flow]. PMID- 2977242 TI - [Open treatment of postoperative peritonitis]. PMID- 2977243 TI - [Surgical methods in intestinal fistulas]. PMID- 2977244 TI - [Complications of the urachus in adults]. AB - The authors observed 7 patients aged from 19 to 74. Five of them were operated upon. One patient died from anaerobic neclostridial phlegmon of the anterior abdominal wall which appeared after an injury of the not noticed urinary duct. Laparotomic accesses passing aside form the area of possible localization of the urinary duct were more justified. When operating on the paraumbilical area it is expedient to make a revision for the detection of the urachus and associated abnormalities on the anterior abdominal wall. PMID- 2977245 TI - [In memory of Il'ia Ivanovich Deriabin]. PMID- 2977246 TI - [In memory of Aleksei Grigor'evich Shapkin]. PMID- 2977248 TI - [The use of computer technology in instructing students in the military department of a medical institute]. PMID- 2977247 TI - [Various problems of epidemiology and prevention of suppurative-inflammatory complications in surgery]. AB - An epidemiological analysis of pyo-inflammatory diseases (PID) in surgical department of multiprofile hospitals has been made. PIDs were diagnozed in 16.2% of patients operated upon. The observation of fluctuations of the level of PID with regard for the correlation of severe and "minor" forms and terms of their appearance allows to make a conclusion of the probable place of infection and may serve an indicator of epidemiological situation. The application of film-forming drug "Lyfuzol" for closing operative wounds has decreased the incidence of PID in catarrhal and phlegmonous appendicitis 2.6 times. PMID- 2977249 TI - [Piriformis syndrome. A contribution to the differential diagnosis of lumbago and coccygodynia]. AB - The piriformis syndrome is characterized by pain in the buttock radiating to the leg and to the coccydeal region. It is an isthmus syndrome of the N. ischiadicus and/or N. cutaneous femoris posterior. The most frequent causes are blunt traumas of the buttocks and spasms of the N. piriformis. More often than not the piriformis syndrome is the cause of a "postlaminectomy syndrome" or of a coccygodynia. The diagnosis can be established by a clinical examination with sufficient reliability. PMID- 2977250 TI - Crossed transvertebral puncture to block spinal ganglion in treatment of pain. AB - A simple and highly efficient percutaneous method eliminating the spinal ganglion and its posterior spinal cord root in treating metameric pain is described. Monitored by X-ray the needle crosses the vertebral canal passing e.g. from the right into the left intervertebral space. The intervention itself is made by a mesocaine alcohol block. The first experience with 12 patients is favourable in postdiscotomic syndromes or lumbar and causalgic pains in lower extremities having no mechanical cause discovered by CT. PMID- 2977251 TI - [Calcinosis intervertebralis of children, clinical and radiologic picture, especially of the late phase]. PMID- 2977252 TI - Importance of antibiotics for the prevention of postoperative spondylodiscitis. PMID- 2977253 TI - Radiodiagnostic signs and clinical symptoms of postoperative spondylodiscitis. PMID- 2977254 TI - Clinical evaluation of autonomic (interoreflex) heart rate regulation. Application to diagnosis of autonomic neuropathy. PMID- 2977255 TI - On acute effects of ethanol (0.5 g/kg) in the course of one hour on the blood pressure, the body temperature and the heart rate in man. PMID- 2977256 TI - Effect of stress and ethanol on reproduction of conditioned food motor reflexes in cats. PMID- 2977257 TI - On acute effects of some drugs on the higher nervous activity in man. The acute effects of ethanol (0.5 g/kg) in the course of one hour. Part LVII. PMID- 2977258 TI - Retest reliability of the paired-association learning indices. PMID- 2977259 TI - Histioceptive-neurotrophic system in amyotrophic lateral sclerosis. PMID- 2977260 TI - Effects of microtubular transport blockade in peripheral nerve axons on chronic unallayable root pain. PMID- 2977261 TI - [Clinical efficacy of long-term treatment with LH-RH analogue, ICI 118630 (Zoladex), in prostatic cancer patients. The Zoladex Multicenter Study Group]. AB - The Zoladex (ICI 118,630) multicenter trial included 149 patients with stage B +o D prostatic cancer recruited from 1984 to date. Of them 53 clinical responders on the treatment for 40 weeks or longer were subjected to assessment of antitumour response, overall subjective response, endocrinological response, safety and usefulness. The responders consistently showed a clinical response as evidenced by antitumour response in 32 of 50 patients (64.0%) at week 12 and in 35 of 51 (68.6%) at week 40, and overall subjective response in 42 out of 47 patients (89.4%) at both week 12 and week 40. Endocrinologically, all of the 47 eligible patients maintained a 40-week or longer response. Adverse reactions were observed in 19 out of 73 patients (26.0%), subdivided by the time of occurrence as 15/73 (20.5%) up to week 12, 3/71 (4.6%) between week 12 and week 40, and 5/53 (9.4%) in and after week 40. No patient required the discontinuance of treatment. Usefulness of the drug was observed in 51 out of 52 patients (98.1%). The results indicated that Zoladex in a once monthly regimen may be of great advantage to elderly patients with prostatic cancer, and allow an improved patient compliance: Zoladex may not only produce clinical remission but also improve the quality of life. PMID- 2977262 TI - LH-RH agonist, Zoladex (Goserelin), depot formulation in the treatment of prostatic cancer. Randomized dose-finding trial in Japan. AB - Ninety patients with advanced prostatic cancer (15 with stage B, 23 with stage C, and 52 with stage D) were randomized to receive 0.9, 1.8, or 3.6 mg, respectively, of Zoladex depot subcutaneous injection every 28 days for 12 weeks. The serum levels of LH, FSH, and testosterone were elevated after the first injection, and followed by a significant decrease. The suppression of testosterone levels in the blood to castrate levels was observed in all patients except two treated with 0.9 mg. Objective response (CR and PR) was seen in 63.6% (0.9 mg), 47.8% (1.8 mg), and 68% (3.6 mg) of patients according to the Japanese Prostatic Group Criteria. Subjective improvement (performance status, analgesic consumption) was also observed in 75-88% of patients but without a statistically significant difference between each dose group. Only minor adverse effects were found during the treatments. The drug was detected dose dependently in the blood by radioimmunoassay. These results suggest that endocrine therapy with Zoladex depot in doses of 3.6 mg subcutaneously every 4 weeks is a useful alternative to surgical castration in patients with prostatic cancer. PMID- 2977263 TI - Zoladex as primary therapy in advanced prostatic cancer. A French cooperative trial. AB - From April 1984 to May 1986, 129 patients with prostate cancer entered a prospective trial with a new LH-RH agonist, Zoladex. Mean age was 72 years (range of 45-94 years) and, in most cases, patients had metastatic disease, not previously treated by chemotherapy or hormone therapy. Patients received a monthly injection of 3.6 mg. Serum testosterone was lowered into the range of castrate levels after 4 weeks of treatment. In 105 evaluable patients at 3 months, a 65% partial response (PR) rate was observed, with 11% stable and 24% progressive disease. Median time to progression was 37 weeks. Analysis of objective criteria revealed 30% PR for prostate volume and 51% CR-PR for prostatic acid phosphatases. Seventeen percent of lytic metastases had recalcified. One hundred twenty-nine patients were evaluable for toxicity. Endocrinological side effects were common: decrease in libido, 92%; impotence, 86%; hot flushes, 48%; and breast swelling or tenderness, 9%. Nonendocrinologic side effects were rare. The treatment is generally well accepted by patients owing to the convenient depot formulation and to the minor side effects. PMID- 2977264 TI - Long-term results of treating advanced prostatic cancer with the LH-RH analogue Zoladex. AB - Fifty-six patients with previously untreated advanced prostatic cancer were treated with the LH-RH agonist Zoladex. Patients have been followed for between 24 and 39 months. The response rate and duration of response have been assessed and related to the degree of differentiation of the primary tumor. All of the patients with well-differentiated tumors responded symptomatically, while the comparable figure for patients with poorly differentiated tumors was 67%. The better differentiated tumors tended to respond longer than their poorly differentiated counterparts (18.75 months compared to 15.9 months, respectively). Overall survival was worse in patients having tumors of higher grade and the mean survival after relapse was shorter. The results confirm the unfavorable prognostic significance of poor histological grade in prostatic cancer and indicate that response to treatment, duration of response, and time to death following relapse are also adversely influenced. PMID- 2977265 TI - Clinical study of an LH-RH agonist (ICI 118.630, Zoladex) in the treatment of prostatic cancer. AB - Eighty patients with prostatic cancer have been treated with an LH-RH analogue (Zoladex). Ten had no metastasis, and hormone therapy was used as an induction treatment before curative radiotherapy. The others had metastatic disease and, in some cases, had already received some form of endocrine therapy. Patients received a monthly injection of Zoladex (3.6 mg). No progressive disease was noted among patients with nonmetastatic tumors; of the patients with metastases, those who were previously untreated had a higher response rate (14.8% complete response) and longer progression-free and overall survival. Toxicity was mild in spite of two cases of disease flare. PMID- 2977266 TI - LH-RH analogue treatment for advanced prostate cancer. AB - The trial drug was ICI 118.630 (Zoladex). Inclusion criteria were histologically confirmed advanced prostate cancer (T greater than 2 or N+ or M+), life expectancy greater than 3 months, and no previous radiotherapy, orchiectomy, or chemotherapy. Treatment started in November 1984; 30 patients were recruited. The period of treatment ranged from 6 to 144 weeks (median of 59.5 weeks). One patient died after 6 weeks of rapidly progressive renal failure. Data were updated to the end of August 1987. The mean age was 67.9 years (53-83 years). Subjective response was evaluated by a mean symptoms score (using daytime micturition, nocturia, dysuria, hesitancy, and flow) and a score of three different items: patients' activity, bone pain, and use of analgesics. Only 7.1% of the patients showed a permanent positive response. Four different objective responses (complete, partial, stable disease, and progression) were possible after evaluating the T category, tumor dimensions, metastases, and prostatic acid phosphatase. Testosterone (T) and plasmatic LH levels rose after administration: T dropped below the castration level (1 ng/ml) within a few days and remained constantly low. The rate of progressive disease was 27.6%; disease control was possible in 72.4% of the patients (PR or SD). PMID- 2977267 TI - Zoladex treatment of symptomatic prostatic carcinoma. AB - The depot LH-RH agonist Zoladex was used to treat 38 patients with previously untreated symptomatic stage D2 prostate carcinoma. Side effects were minimal and patient acceptability excellent, although temporary tumor flare occurred in 11% of patients. Eighty-four percent experienced subjective improvement and 87% had objective evidence of initial disease stabilization or remission lasting 3 months. Serum levels of gonadotrophin and free testosterone as well as androgens of adrenal origin fell significantly with treatment. Long-term survival to date appears at least as good as that described for conventional endocrine therapy. PMID- 2977268 TI - Simultaneous administration of a luteinizing hormone releasing hormone agonist and diethylstilbestrol in the initial treatment of prostatic cancer. AB - It is well known that the first administration of luteinizing hormone releasing hormone (LH-RH) analogues induces an initial increase of luteinizing hormone (LH) release and a subsequent rise in testosterone (T). This rise is maximal after 3-4 days and is followed by a steady fall of LH and T. Several investigators have reported flare-up symptoms in patients with advanced prostatic cancer, associated with the rise of T, when treated by LH-RH analogue alone. In order to prevent these flare-up symptoms, we treated 20 patients with advanced prostatic cancer with an association of a LH-RH analogue (Zoladex Depot) and diethylstilbestrol (DES) 1 mg/day. DES was given for 14 days, starting 7 days before the first Zoladex Depot injection. T fell to near castrate levels within a few days, but rose again 3-4 days after the administration of the LH-RH analogue to pretreatment values before returning to castrate levels. No clinical flare-up manifestations were recorded. We conclude that combination treatment with DES can prevent the flare-up symptoms induced by LH-RH analogue in patients with prostatic cancer. PMID- 2977269 TI - Long-acting (depot) D-TRP-6 LH-RH (Decapeptyl) in prostate cancer. An Italian multicentric trial. AB - Ninety-five patients with stage C (C1 + C2) or D (D1 + D2) prostatic carcinoma were treated with the long-acting formulation of D-TRP-6 LH-RH (Decapeptyl) for up to 39 months. Of 88 patients evaluable for response, about one-half showed an objective response. In most cases, subjective improvement with relief of bone pain and/or urinary symptoms was obtained. Five patients claimed a mild increase in bone pain and one patient a slight worsening of dysuria following the first injection. Median progression-free survival was 13.1 and 16.4 months in patients with stage D2 and D1, respectively. Median survival in stage D2 patients was 27.6 months. These results indicate that the depot formulation of D-TRP-6 LH-RH offers an effective therapeutic alternative for patients with advanced prostatic cancer. PMID- 2977270 TI - Hormonal therapy trials in prostatic cancer. An EORTC Genitourinary Group study. AB - In this paper the authors review the completed and current EORTC Genitourinary Group trials for metastatic carcinoma of the prostate. In terms of time to progression and length of survival, there is no significant difference between any of the effective endocrine treatments that have been studied. The statistical analysis of the different variables used in trials 30,761 and 30,762 determines three risk groups of patients. There is no need for very special laboratory investigations to establish a prognosis. Orchiectomy is the cheapest and safest endocrine treatment in metastatic carcinoma of the prostate. PMID- 2977271 TI - How would you like to have an orchidectomy for advanced prostatic cancer? AB - One hundred thirteen patients were randomized for the study. There was no significant difference between the groups for response or survival. The incidence of side effects related to the reduction of testosterone was similar in both groups. Disease "flare" was seen in three patients treated with long-acting D-Trp 6 LH-RH. All symptoms resolved by the end of 8 weeks. There was less psychological morbidity in the D-Trp-6 LH-RH group but the difference did not reach statistical significance. Our results indicate that long-acting D-Trp-6 LH RH offers a safe and effective alternative to surgical orchidectomy. PMID- 2977272 TI - U.K. trials of treatment for M1 prostatic cancer. The LH-RH analogue Zoladex vs. orchidectomy. AB - A randomized, controlled trial of the treatment of metastatic prostatic cancer using either the LH-RH analogue Zoladex or orchidectomy is reported. The trial was conducted in the U.K. and Ireland and involved participants in 17 centers. The design of the trial is outlined and the response criteria discussed. The results show that the patient characteristics in the two treatment groups were comparable upon entry. Repeated administration of Zoladex every 28 days has been shown to be as effective as orchidectomy in lowering serum testosterone to castrate levels. The subjective and objective response rates were similar, as were the duration of response, time to treatment failure, and survival rates. The withdrawals from the trial and adverse reactions are discussed. These results, 10 months after the closure of recruitment to the trial, seem to indicate that Zoladex and orchidectomy are equivalent treatments and that Zoladex seems to be a truly medical alternative to surgical castration. Finally, other ongoing U.K. trials of the treatment of advanced prostate cancer are outlined. PMID- 2977273 TI - Interim report of a randomized trial comparing Zoladex 3.6 mg depot with diethylstilbestrol 3 mg/day in advanced prostate cancer. The West Midlands Urology Research Group. AB - A study comparing Zoladex 3.6 mg depot with diethylstilbestrol (DES) 3 mg/day was initiated in August 1985. One hundred ninety-three patients with histologically confirmed prostate cancer T3/4 or M1 have been randomized up to 31 March 1987: 95 to Zoladex, 98 to DES. No patient had received prior systemic therapy. There is no bias in the treatment groups in terms of baseline characteristics. Median follow-up is 11 months, and the response rate at 12 months from randomization (CR + PR) for Zoladex is 70 +/- 9.4% and 50 +/- 10.1% for DES. Median time to best response is 6 months for Zoladex and 12 months for DES (using the Kaplan-Meier life table method). Subjective responses are 56 +/- 10.2% for Zoladex and 44 +/- 10% for DES. Five increases in bone pain were found after the first Zoladex treatment, as well as one increased ureteric obstruction. None required treatment withdrawal. Seventeen patients on DES were withdrawn due to adverse reaction (chi 2 = 4.33, 1df, p less than 0.05). Overall survival at 31 March 1987 is 84% for the Zoladex group and 78% for the DES group. This study has shown that Zoladex is superior to DES in achieving early tumor response in advanced prostate cancer, without causing serious complications warranting withdrawal of treatment. PMID- 2977274 TI - [Analysis of the sequelae of induced abortion (based on the clinical data of the V.I. Lenin Kazan Institute for the Training of Physicians)]. PMID- 2977275 TI - [The impossible handicap. Analysis of the concept of handicap in the orientation law of 30 june 1975. II]. PMID- 2977276 TI - [Towards an attempt at biological differentiation of endogenous and exogenous depression: a study of beta-endorphins and other parameters (prolactin, growth hormone and dexamethasone repression test) in a cohort of ambulatory depressed patients]. PMID- 2977277 TI - Hypersensitivity to Shigella outer membrane proteins in mice. AB - Mice sensitized with living Sh. flexneri 3a or with outer membrane proteins (OMP) derived from these bacteria showed Arthus and delayed type hypersensitivity (DTH) when injected in the footpad with OMP. The delayed reactivity to OMP was transferable to normal mice by spleen cells obtained from donor animals which were previously sensitized with living bacteria. PMID- 2977278 TI - Altered expression of chitin synthetase activity and biochemical changes in the cell wall in a developmental mutant of Phycomyces. AB - The Phycomyces developmental mutant S356 elaborates spores which show a much poorer viability and a higher affinity for Calcofluor White than the wild-type spores. Protease-activated extracts of the mutant spores showed higher levels of chitin synthetase activity than the parental strain-derived spores. High levels of enzyme activity in the mutant extracts, but not in the corresponding wild-type extracts, could be detected in the absence of an exogenous protease. The high basal active chitin synthetase is not the result of activation by endogeneous proteases during cell breakage since protease inhibitors did not reduce, but rather increased, the activity levels. The analysis of cell wall composition in the mutant spores revealed significant changes in the proportion of uronic acids and protein but not in chitin. The mutant phenotype is discussed in relation to the developmental stage at which the alterations connected with cell wall metabolism occurred. PMID- 2977279 TI - Tourette syndrome. PMID- 2977280 TI - Reduction of elevated CSF beta-endorphin by fenfluramine in infantile autism. AB - Fenfluramine therapy has been reported to improve behavior in infantile autism and has been associated with a decrease in abnormally increased blood serotonin content. The primary central effect has not been proved to be serotonergic. Beta endorphin is involved in the anorexic effect of fenfluramine and may play a role in autism. Nine children with infantile autism were treated with fenfluramine in double-blind, placebo-crossover design. Transient anorexia was the only adverse effect. Autistic behavior was reported to improve in three patients, but objective psychometric tests were unchanged. Beta-endorphin-like immunoreactivity was determined in lumbar cerebrospinal fluid of patients during and before or after treatment with fenfluramine and then was compared to normal controls. Beta endorphin was elevated significantly in the baseline autistic group (p less than .005) and was reduced toward control values during fenfluramine treatment. The results are consistent with a role for beta-endorphin in infantile autism and in the mechanism of fenfluramine treatment. PMID- 2977281 TI - Rehabilitation of communication impairment in dystonia musculorum deformans. AB - Augmentative and alternative communication (AAC) aids were used in three young, intellectually normal patients with dystonia musculorum deformans (DMD) who had severe speech and writing impediments. These aids included speech therapy, communication boards, and voice synthesizers for verbal communication and typewriters, memowriters, and computer software and printers for written communication. At times customized accessing was needed which required specific adaptive modifications. Implementation of the AAC aids system was determined effective for DMD patients in view of the intellect-sparing nature of the disorder. Improvement was hampered by the progressive nature of the disease and by the emotional stress of accepting the long-term use of AAC. Correct and early diagnosis of communication impediments are crucial for the appropriate AAC aids prescription and implementation. An AAC protocol is suggested to meet the special communication needs of DMD patients. PMID- 2977282 TI - Recurrent encephalomyelitis associated with incontinentia pigmenti. AB - A 4-year-old girl developed progressive obtundation following an upper respiratory tract illness. Physical, cerebrospinal fluid, computed tomographic, electroencephalographic, and evoked response findings were compatible with disseminated encephalomyelitis. Skin lesions indicative of incontinentia pigmenti were confirmed by biopsy. Transient loss of suppressor T cells was observed. Prior history revealed that at 6 months of age a similar episode of acute central nervous system deterioration had occurred. Recurrent encephalomyelitis does occur with incontinentia pigmenti. Transient loss of suppressor T cells suggests that this is an immune-mediated process. PMID- 2977283 TI - Huntington disease: finding the gene and after. AB - Huntington disease is an autosomal dominant disorder that usually begins in mid life and is characterized by progressive choreiform movements and dementia. Approximately 5% of patients develop symptoms prior to 14 years of age. In most juvenile cases, the gene is transmitted from the father. In children the clinical course is marked by mental deterioration or behavioral abnormalities, gait disturbances usually the consequence of rigidity, cerebellar signs, and seizures. The pathologic findings are highlighted by atrophy of the caudate. Atrophy also is observed on brain imaging, while positron emission tomography demonstrates marked caudate hypometabolism which antedates the appearance of the clinical disease. Cell death in the striatum primarily affects medium and small GABA containing neurons, representing the striatal output projections. Somatostatin containing neurons and cholinergic neurons are spared. The gene for Huntington disease has been localized in close proximity to the tip of the short arm of chromosome 4. The gene product and the manner by which it induces selective cell death is still unknown but should become evident in the near future. PMID- 2977284 TI - [Intracranial venous blood flow velocities in infants]. PMID- 2977285 TI - Venous responsiveness to atrial natriuretic factor in man. AB - The venorelaxant effect of atrial natriuretic factor in man was studied using the dorsal hand vein technique. Infusion of met-ANF to preconstricted veins at doses up to 240 ng min-1 in 11 healthy male subjects caused only minimal venorelaxation. Atrial natriuretic factor is unlikely to have a significant venorelaxant effect at physiological doses in man. PMID- 2977286 TI - UV-induced vanadate-dependent modification and cleavage of skeletal myosin subfragment 1 heavy chain. 1. Evidence for active site modification. AB - Ultraviolet irradiation above 300 nm of the stable MgADP-orthovanadate (Vi) myosin subfragment 1 (S1) complex resulted in covalent modification of the S1 and in the rapid release of trapped MgADP and Vi. This photomodified S1 had Ca2+ATPase activity 4-5-fold higher than that of the non-irradiated control S1, while the K+EDTA-ATPase activity was below 10% of controls. There was a linear correlation between the activation of the Ca2+ATPase and the release of both ADP and Vi with irradiation time. Analysis of the total number of thiols and the ability of photomodified S1 to retrap MgADP by cross-linking SH1 and SH2 with various bifunctional thiol reagents indicated that the photomodification did not involve these reactive thiols. Irradiation of the S1-MgADP-Vi complex caused a large increase in absorbance of the enzyme at 270 nm which was correlated with the release of Vi from the active site, suggesting an aromatic amino acid(s) was (were) involved. However, analysis by three different methods showed no loss of tryptophan. All the irradiation-dependent phenomena could be prevented by replacing Mg2+ with either Co2+, Mn2+, or Ni2+. Unlike previous irradiation studies of Vi-dynein complexes [Lee-Eiford, A., Ow, R. A., & Gibbons, I. R. (1986) J. Biol. Chem. 261, 2337-2342], no peptide bonds were cleaved in photomodified S1. Photomodified S1 was able to retrap MgADP-Vi at levels similar to unmodified S1. Upon irradiation of the photomodified S1-MgADP-Vi complex, MgADP and Vi were again released from the active site, resulting in heavy chain cleavage to form NH2-terminal 21-kDa and COOH-terminal 74-kDa peptides. All evidence indicates that this new photomodification and subsequent chain cleavage occur specifically at the active site. PMID- 2977287 TI - Solubilization and reconstitution of the D-1 dopamine receptor: potentiation of the agonist high-affinity state of the receptor. AB - The D-1 dopamine receptor was extracted from rat striatal membranes with sodium cholate and NaCl in the presence of a specific agonist and phospholipids. The soluble receptor then was reconstituted into phospholipid vesicles by further addition of phospholipids prior to detergent removal. Of the total membrane receptors, up to 48% were extracted and 36% were reconstituted into phospholipid vesicles. Yields were greatly reduced if the agonist was omitted or replaced with an antagonist. The solubilized and reconstituted D-1 receptors retained the pharmacological properties of the membrane-bound receptors, including the ability to discriminate between active and inactive enantiomers of specific agonists and antagonists. In this regard, the affinity of the reconstituted receptors for the D-1 specific antagonist 125I SCH 23982 was similar to that of the membrane-bound receptors with a Kd of 1.5 nM. Both the soluble and reconstituted forms of the D 1 receptor exhibited two affinity states for the D-1 specific agonist SK&F R 38393. In contrast to the low proportion of the receptors that had a high affinity for the agonists in striatal membranes (less than 6%), there was a dramatic increase following solubilization (22%) and reconstitution (40%). Similar results were obtained by using dopamine; the proportion of high-affinity sites increased from 4% (membrane-bound) to 48% (reconstituted) of the total receptor population. These high-affinity sites were coupled to G proteins, as guanyl nucleotides completely abolished them. Addition of guanyl nucleotides prior to solubilization or to reconstitution, however, had no effect on the subsequent yield of the reconstituted receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977288 TI - High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 3. Effect of electrical field shape. AB - The resolution of pulsed-field gel electrophoresis is dramatically affected by the number and configuration of the electrodes used, because these alter the shape of the applied electrical fields. Here we present calculations and experiments on the effect of electrode position in one of the most commonly used pulsed-field gel electrophoresis configurations. The goal was to explore which aspects of the electrical field shape correlate with improved electrophoretic resolution. The most critical variable appears to be the angle between the alternate electrical fields. The most effective electrode configurations yield angles of more than 110 degrees. A continually increasing angle between the fields produces band sharpening that greatly enhances the resolution. PMID- 2977289 TI - High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 4. Influence of DNA topology. AB - Pulsed-field gel electrophoresis is a powerful technique for the fractionation of linear DNA molecules with sizes above 50 kilobase pairs (kb). Here it is demonstrated that this technique is also effective for separating smaller DNAs including linear, circular, and supercoiled species. The mobilities of linear DNAs larger than 8 kb can be modulated by pulse times between 0.1 and 100 s. The mobility of supercoiled DNA molecules up to 16 kb is generally unaffected by these pulse times except that 10-s pulse times cause a small but distinct increase in the mobility. The general insensitivity of small supercoiled DNAs to pulse time presumably occurs because these species reorient so rapidly that they spend most of their time undergoing conventional electrophoresis. However, the mobilities of larger supercoiled DNAs are affected by pulse times of less than 1 s, and at 0.1 s the molecules are better resolved by pulsed electrophoresis than by ordinary electrophoresis. The mobility of 3-19 kb nicked and relaxed circular DNA molecules is also affected by pulse time but in a complex way. PMID- 2977290 TI - The biosynthesis of highly branched N-glycans: studies on the sequential pathway and functional role of N-acetylglucosaminyltransferases I, II, III, IV, V and VI. AB - At least 6 N-acetylglucosaminyltransferases (GlcNAc-T I, II, III, IV, V and VI) are involved in initiating the synthesis of the various branches found in complex asparagine-linked oligosaccharides (N-glycans), as indicated below: GlcNAc beta 1 6 GlcNAc-T V GlcNAc beta 1-4 GlcNAc-T VI GlcNAc beta 1-2Man alpha 1-6 GlcNAc-T II GlcNAc beta 1-4Man beta 1-4-R GlcNAc T III GlcNAc beta 1-4Man alpha 1-3 GlcNAc-T IV GlcNAc beta 1-2 GlcNAc-T I where R is GlcNAc beta 1-4(+/- Fuc alpha 1 6)GlcNAcAsn-X. HPLC was used to study the substrate specificities of these GlcNAc T and the sequential pathways involved in the biosynthesis of highly branched N glycans in hen oviduct (I. Brockhausen, J.P. Carver and H. Schachter (1988) Biochem. Cell Biol. 66, 1134-1151). The following sequential rules have been established: GlcNAc-T I must act before GlcNAc-T II, III and IV; GlcNAc-T II, IV and V cannot act after GlcNAc-T III, i.e., on bisected substrates; GlcNAc-T VI can act on both bisected and non-bisected substrates; both Glc-NAc-T I and II must act before GlcNAc-T V and VI; GlcNAc-T V cannot act after GlcNAc-T VI. GlcNAc-T V is the only enzyme among the 6 transferases cited above which can be essayed in the absence of Mn2+. In studies on the possible functional role of N glycan branching, we have measured GlcNAc-T III in pre-neoplastic rat liver nodules (S. Narasimhan, H. Schachter and S. Rajalakshmi (1988) J. Biol. Chem. 263, 1273-1281). The nodules were initiated by administration of a single dose of carcinogen 1,2-dimethyl-hydrazine.2 HCl 18 h after partial hepatectomy and promoted by feeding a diet supplemented with 1% orotic acid for 32-40 weeks. The nodules had significant GlcNAc-T III activity (1.2-2.2 nmol/h/mg), whereas the surrounding liver, regenerating liver 24 h after partial hepatectomy and control liver from normal rats had negligible activity (0.02-0.03 nmol/h/mg). These results suggest that GlcNAc-T III is induced at the pre-neoplastic stage in liver carcinogenesis and are consistent with the reported presence of bisecting GlcNAc residues in N-glycans from rat and human hepatoma gamma-glutamyl transpeptidase and their absence in enzyme from normal liver of rats and humans (A. Kobata and K. Yamashita (1984) Pure Appl. Chem. 56, 821-832). PMID- 2977291 TI - Natural killer cells discriminate between high mannose- and complex-type asparagine-linked oligosaccharides. AB - Chinese hamster ovary cell lines with different types of N-linked oligosaccharides were tested as targets for control and lymphokine treated natural killer (NK) cells. The targets tested were parent cells, Lec1 mutants and Lec4 mutants. Due to an apparent defect in GlcNAc transferase V, Lec4 cells produce complex-type N-linked oligosaccharides devoid of GlcNAc beta(1-6) linked branches. Lec1 cells form only high mannose-type N-linked oligosaccharides because they lack GlcNAc transferase I activity. Lec1 cells are very sensitive to lysis by beta-interferon treated human NK cells, but both parent and Lec4 cells are resistant to NK lysis. The ability to discriminate between parent and Lec1 targets was demonstrated with untreated control effectors as well as those which were pretreated with either beta-interferon, gamma-interferon or interleukin-2. Both control and lymphokine-boosted NK cells exhibit much greater lytic activity against targets having only high mannose-type N-linked oligosaccharides. Five oligosaccharide structures resembling those found on N-linked glycoproteins were tested for their ability to block NK lysis of Lec1 targets. Only the high mannose type glycopeptide from 7S soybean glycoprotein was inhibitory in the mu molar range. At the same concentration, none of the complex-type oligosaccharides had any effect on lytic activity. The results suggest that a high mannose-type N linked oligosaccharides is recognized at some step in NK cell-mediated lysis. PMID- 2977292 TI - Modulatory effects of beta-endorphin on interferon-gamma production by cultured peripheral blood mononuclear cells: heterogeneity among donors and the influence of culture medium. AB - The stress-responsive neuropeptide beta-endorphin (beta-END) has been shown to either enhance or suppress interferon-gamma (IFN-gamma) production by mitogen stimulated peripheral blood mononuclear cells (PBMNC) in vitro. In this study, we investigated whether donor selection and the medium used for cell culture influenced the modulatory effect of beta-END on the IFN-gamma production by PBMNC. Considerable variation of the effect of beta-END on IFN-gamma production was observed in individuals who underwent multiple testing. Suppression of IFN gamma was significantly greater in 16 male donors (29 +/- 6.0%) compared to 8 female donors (1.5 +/- 6.4%) when PBMNC were preincubated for 3 h with beta-END followed by concanavalin A stimulation for 72 h in medium containing 20% fetal bovine serum (FBS). Lowering the concentration of FBS resulted in enhanced production of IFN-gamma by PBMNC exposed to beta-END. Arachidonic acid or beta END added separately to medium containing 3% autologous serum produced no suppression of IFN-gamma, but when added together resulted in significant suppression. This observation provides support for the hypothesis that beta-END achieves its suppressive effect on IFN-gamma via a mechanism involving arachidonic acid metabolism. We conclude that the modulatory effect of beta-END on IFN-gamma production by PBMNC is both donor and medium dependent. PMID- 2977293 TI - Septal Q wave response to exercise before and after percutaneous transluminal coronary angioplasty. PMID- 2977294 TI - Reduction of the adherence of Streptococcus sobrinus insoluble alpha-D-glucan by endo-(1----3)-alpha-D-glucanase. AB - Insoluble alpha-D-glucan, previously formed on a glass surface from sucrose by the action of cell-free D-glucosyltransferases of Streptococcus sobrinus OMZ176, was significantly removed by a purified preparation of endo-(1----3)-alpha-D glucanase (mutanase) from a strain of Pseudomonas sp. Almost complete dissociation of adherent glucan occurred at the highest enzyme concentration (40 mU/mL) tested. Synthesis and de novo adherence on glass of the glucan was markedly inhibited by the presence of mutanase, even at low concentrations (4 mU/mL or less). When compared to native glucan, the mutanase-modified glucan samples (a) contained lower proportion of D-(1----3) linkages; (b) showed lower susceptibility to mutanase and higher susceptibility to (1----6)-alpha-D glucanase (dextranase); (c) contained larger amounts of low-molecular-weight fractions; (d) had lower intrinsic viscosities; (e) showed higher S. sobrinus cell-agglutinating activities; and (f) consisted of looser entwinement of coalescent single-stranded fibrils (a major component) and shorter double stranded fibrils (a minor one). PMID- 2977295 TI - Maintenance of heparan sulfate structure throughout evolution: chemical and enzymic degradation, and 13C-n.m.r.-spectral evidence. PMID- 2977297 TI - Decrease of serum buffering capacity associated with malignant neoplasms. AB - We have developed a new tumor marker based on the finding that cancerous sera suppress glucose utilization by cultured macrophages. Phosphofructokinase (PFK) was identified as susceptible enzyme [Naknamura et al.: JNCI 1986; 77 (in press)]. We found a high correlation between PFK inhibition and the lower buffering capacity of cancerous sera against an ATP aqueous solution and diluted acidic solutions. Sera from healthy donors changed to the PFK-suppressive type when exposed to lactate, even when followed by elimination of lactate. Lactate in cancerous sera was 1.6 times more than in normal sera. Sera from patients who tend to show acidosis, such as those with diabetes mellitus or chronic renal failure and pregnant women, also showed a lower buffering capacity as well as a higher inhibition of PFK. Although the pH method is simpler, we recommend the application of the PFK inhibition test, because this enzyme inhibition is not simply composed of a lower buffering capacity of cancerous sera, but partially of an inactivation of the enzyme through oxidation. PMID- 2977296 TI - Natural occurrence and clastogenic effects of nivalenol, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl-deoxynivalenol, and zearalenone in corn from a high risk area of esophageal cancer. AB - This is the first report of the natural coexistence of a group of Fusarium mycotoxins (nivalenol [NIV], deoxynivalenol [DON], 3-acetyl-deoxynivalenol [3 ADON], 15-acetyl-deoxynivalenol [15-ADON], and zearalenone [ZEN]) in corn from Linxian, China, an area with a high risk of esophageal cancer. Using thin layer chromatography (TLC), high pressure liquid chromatography (HPLC), and gas chromatography (GC), 107 corn samples from Linxian were analyzed. The average levels of NIV and DON were 757 +/- 707 (54-2,760) ng/g and 5,376 +/- 4,460 (360 12,670) ng/g, respectively, with 100% positivity in 24 corn samples consumed as staple food by esophageal cancer patients and their families. Other corn samples collected from five villages in Linxian at different seasons in 1984-1986 also revealed high levels of NIV and DON contamination, with 100% positivity, suggesting that they are consistently and widely present in corn in that area. Levels of 3-ADON and 15-ADON in Linxian corn were 113 +/- 57 and 495 +/- 538 ng/g, respectively. Crude extracts of corn samples collected from esophageal cancer patients' families and the HPLC-purified NIV and DON fractions induced significant chromosome aberrations in V79 cells. Pure toxins of NIV, DON, T-2, and 3-ADON also induced chromosome aberrations in V79 cells at very low concentrations (ng levels/ml medium). Cytotoxic effects were observed at slightly higher concentrations. The levels and kinds of trichothecenes were in positive co relation with the incidence of esophageal cancer. The data suggest that trichothecenes in food may possibly be associated with esophagitis and esophageal cancer in Linxian. PMID- 2977298 TI - Characterization of two similar differential tumor markers based on phosphofructokinase activity arising from the influence of cancer patient serum. AB - The activity of phosphofructokinase (ATP: D-fructose-6-phosphate 1 phosphotransferase, EC.2.7.1.11) was found to decrease through the influence of sera from cancer patients. Two similar differential methods were developed using phosphofructokinase (PFK) as the indicator enzyme. In the original "acidic ATP method" (AAM), an acidic ATP-aqueous solution is mixed with a PFK-serum mixture following incubation at room temperature for 9 min prior to injecting a reaction cocktail without ATP into the latter. In another "two-step method" (TSM), the reaction cocktail including ATP is added to a mixture of PFK and serum after its incubation for 60-90 min at 37 degrees C. The reaction mechanism of AAM is as follows: The buffering capacity of sera decreases with the progress of malignant tumors, whose physiology is termed subdetectable acidosis, but more recently designated as hydrogen ionic stress. In contrast, TSM occurs through an increase in serum oxidation activity, causing reversible oxidation of PFK and subsequent decrease in PFK activity. Such physiology is presently termed oxidative stress [Sies S: Oxidative Stress. London: Academic Press, 1985] and recognized as an increase in active oxygens, lipid peroxides, and oxidation enzymes in the serum. These findings are based on examination of the effect of ammonium sulfate at various concentrations, dithiothreitol in a PFK suspension, antagonistic reagents, sodium azide, effects of various volumes and dilutions of serum on the PFK activity, and the clinical application of both methods. We conclude that AAM and TSM have entirely different reaction mechanisms and thus different clinical applications. PMID- 2977299 TI - Orthopedic aspects of competitive swimming. AB - Orthopedic problems related to competitive swimming are rarely disabling, but can be problematic in preventing training and competition. Most problems are related to the shoulder and knee. Treatment is primarily nonsurgical and directed at relieving symptoms and allowing the athlete to continue with swimming practice. Treatment aids such as ice packing, anti-inflammatory medications, muscle stimulation and electrogalvanic stimulation, strengthening exercises, and static stretching are encouraged; upper arm bands and patellar-stabilizing supports can be adapted to training routines. PMID- 2977300 TI - [Experimental hypertrophy of the heart and its regression in rats--changes in functional reserve]. PMID- 2977301 TI - Isolation and characterization of a Neurospora glucose-repressible gene. AB - Using differential hybridization, the cDNA copy of a Neurospora gene coding for an abundant glucose-repressible mRNA (grg-1) has been isolated. The cDNA was used to clone the genomic copy, and both were sequenced. The cDNA is nearly full length and contains putative translational start and termination codons. Conceptual translation indicates that grg-1 mRNA could direct the synthesis of a 7,000 molecular weight polypeptide. The genomic clone, contained in an 1,888 bp PvuII fragment, encompasses the entire cDNA as well as 838 bp of 5' and 369 bp of 3' flanking sequence. Comparison of the cDNA and genomic clones revealed the presence of two short introns in potential protein-coding sequences. grg-1 message levels were found to increase within minutes following the onset of glucose deprivation and rise 50 fold during the first 90 min of derepression. PMID- 2977302 TI - Immediate hypersensitivity reactions to penicillin and related antibiotics. PMID- 2977303 TI - Priming effect of a birch pollen season studied with laser Doppler flowmetry in patients with allergic rhinitis. AB - Nasal mucosal provocation tests were done on eight patients with seasonal allergic rhinitis before and after a birch pollen season. The effects on nasal microvascular blood flow were detected by means of laser Doppler flowmetry. The patients reacted to the birch pollen provocation with an increase in blood flow. This increase was greater after the pollen season than before, when the same pollen doses were used, indicating a priming phenomenon of the resistance vessels. PMID- 2977304 TI - Evidence against the central nervous system involved in altered reflex control of sympathetic nerve activity by atrial natriuretic peptide. AB - This study aimed to examine whether atrial natriuretic peptide (ANP) alters arterial baroreceptor control of sympathetic nerve activity (SNA) by acting on the central nervous system (CNS). In rabbits anesthetized with alpha-chloralose, we examined changes in arterial pressure and renal SNA in response to graded electrical stimulation (1.7, 3.3, 5.0 Hz) of aortic depressor nerves during the intravenous (IV) infusion of saline or alpha-human atrial natriuretic peptide (alpha-hANP) (0.1, 1.0 microgram/kg/min) or after the intracerebroventricular (ICV) injections of saline or alpha-hANP (3, 10 micrograms). Sinoaortic denervation and bilateral vagotomy were done before examination in all rabbits to eliminate the influence of the known effects of ANP on arterial and cardiac receptors. IV alpha-hANP decreased arterial pressure but did not alter renal SNA. ICV alpha-hANP did not change arterial pressure or renal SNA. Arterial pressure and renal nerve responses to graded electrical stimulation of aortic depressor nerves were not altered by IV or ICV alpha-hANP. These results suggest that alpha hANP does not modulate control of renal SNA by acting on CNS. PMID- 2977305 TI - Atrial natriuretic peptide, angiotensin, norepinephrine and electrolyte in cerebrospinal fluid of essential hypertension. AB - We determined concentrations of atrial natriuretic peptide (ANP), angiotensin (Ang), norepinephrine (NE) and electrolyte in plasma and cerebrospinal fluid (CSF) to study possible roles of these substances within the brain in human hypertension. Blood and CSF samples were obtained from 10 patients with mild to moderate essential hypertension (EHT) aged 40-65 y and 10 age-matched normotensive subjects (NT) on a regular salt diet (8 g/day). Levels of ANP, NE, Na, K, Ca and Cl in CSF and plasma were comparable between EHT and NT. Plasma renin activity, plasma and CSF Ang II were lower in EHT than NT. CSF Ang III tended to be lower in EHT. There was no correlation between CSF and plasma ANP, or between CSF and plasma Ang II. Our results indicate that CSF levels of ANP may not be altered in middle aged patients with mild to moderate hypertension. It is also suggested that Ang II, NE and sodium in the central nervous system may not have important roles in hypertension of those patients. PMID- 2977306 TI - Tinnitus prevalence in industrial hearing loss compensation claimants. AB - In a study of 3466 claimants for noise-induced occupational hearing loss tinnitus was reported by half of the patients. Its characteristics in this select group were different from tinnitus seen in other groups. Specifically, tinnitus prevalence seemed to be independent of age, state of hearing and duration of noise exposure if greater than 10 years. The reasons for these differences are unclear, though the possibility of financial motivation by some claimants is raised. Further work in this area is urgently required to achieve standardization and objectivity. Being a subjective complaint these aims will be difficult to accomplish. However not until then can a fair handicap scale be developed for workers who develop this disquieting symptom as a result of their occupation. PMID- 2977307 TI - Plasma neuropeptide Y and atrial natriuretic peptide concentrations in man. AB - 1. In order to examine the concentration of neuropeptide Y-like immunoreactivity (NPY-LI) and atrial natriuretic peptide (ANP) in the circulation in man, blood was sampled from the iliac vein, the inferior vena cava, the superior vena cava, the pulmonary artery and the femoral artery in 13 patients undergoing cardiac catheterization. 2. Plasma NPY-LI levels were similar at all points sampled and no arteriovenous differences were found. Plasma ANP concentration in the pulmonary artery was greater than in peripheral venous blood but there was a strong correlation between the two. 3. The concentration of NPY-LI and ANP in peripheral venous blood reflects central venous and arterial concentrations. PMID- 2977308 TI - Cardiac function and cardiovascular hormone balance during hemodialysis with special reference to atrial natriuretic peptide. AB - Echocardiographically determined left ventricular function and cardiovascular hormone balance were assessed before and after hemodialysis in 10 patients who had been on hemodialysis for 4 months to 15 years. Plasma levels of atrial natriuretic peptide (ANP), antidiuretic hormone (ADH), renin activity and aldosterone were determined. All patients had vector- and echocardiographic evidences of slight to moderate left ventricular hypertrophy. The body weight decreased 2.0 kg (3.3 +/- 0.5%) with dialysis. Nine out of ten patients showed a slightly reduced ejection fraction that normalized after dialysis (p less than 0.05). Left atrial and ventricular systolic dimensions were around the upper reference limit before dialysis with a decrease after dialysis (p less than 0.05 and p less than 0.02, respectively). The levels of ANP decreased with dialysis from 2-17 times to 1 to 15 times the upper reference value in nine out of the ten patients. In the whole group the decrease was 117 +/- 35% (p less than 0.005). A significant regression was obtained between percentage decrease of body weight and percentage change of ANP (r = 0.67; p less than 0.05). The plasma concentration of ADH did not change following dialysis but the mean value was significantly higher than the mean value of the reference group of the laboratory (p less than 0.05 before and p less than 0.005 after dialysis). Renin activity and aldosterone levels were low and did not change during dialysis. In conclusion, the slight left ventricular hypertrophy may partly be a response to volume overload with hyperdynamic circulation and partly to metabolically depressed myocardial function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977309 TI - Increased plasma levels of immunoreactive human atrial natriuretic factor in primary IgA nephropathy. AB - The plasma concentration of the immunoreactive (IR) human atrial natriuretic factor (hANF) was measured in 17 patients with primary IgA nephropathy (IgAN) (9 normotensive and 8 hypertensive subjects without impairment of renal function). Furthermore, correlations with the renin-angiotensin II-aldosterone system and hemodynamic alterations were studied. The mean value of IR-hANF was significantly (p less than 0.002) higher in normotensive IgAN patients (68.2 +/- 14.6 pg/ml) than in controls (48.8 +/- 11.5 pg/ml), while it was slightly and not significantly elevated in hypertensive IgAN patients (58.5 +/- 8.4 pg/ml). In the latter the mean plasma renin activity (PRA) was significantly increased (0.92 +/- 0.30 ng/ml/h; p less than 0.002), while in normotensive IgAN patients (0.68 +/- 0.58 ng/ml/h) no difference was observed. Plasma aldosterone levels showed the same behavior pattern as those of PRA. Hemodynamic studies showed that the mean values of the cardiac index (CI) were significantly (p less than 0.002) high in both normotensive (3.55 +/- 0.5 l/min/m2) and hypertensive (3.32 +/- 0.47 l/min/m2) patients, while a significant reduction in the total peripheral resistance index (TPRI) in normotensive (2171 +/- 349 dyn/s/cm-5/m2; p less than 0.02) and a significant increase in hypertensive (2959 +/- 440 dyn/s/cm-5/m2; p less than 0.05) patients were observed. The mean arterial pressure (MAP) had a positive correlation with the TPRI and an inverse correlation with the IR hANF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977310 TI - In vitro study of expression of interleukin-2 receptors in T-lymphocytes from patients with IgA nephropathy. AB - The present study was undertaken to examine the expression of interleukin-2 receptor (IL-2R)/CD25 antigen in cultured T-lymphocyte subsets in IgA nephropathy. Twenty-four IgA nephritic patients, 12 patients with chronic glomerulonephritis (non-IgA nephropathy), and 17 healthy controls were studied in an infection-free interval. T-cell subsets in peripheral blood mononuclear cells (PBMC) and activated T-lymphocyte subsets expressing IL-2R were determined by double immunofluorescence staining with fluorochromes conjugated to monoclonal antibodies against T-helper/inducer (CD4+) cell, T-suppressor/cytotoxic (CD8+) cell, B (CD20+) lymphocytes, and IL-2R. The percentages of CD4+ and CD8+ lymphocytes, CD4/CD8 ratio, and total activated lymphocytes (with IL-2R/CD25 antigen) did not differ between the IgA nephritic patients, patients with chronic glomerulonephritis, and healthy controls in freshly isolated, unstimulated lymphocytes or PBMC cultured with pokeweed mitogen. Following pokeweed mitogen stimulation for 5 days, 17.3 +/- 10.3%, 16.6 +/- 8.4%, and 16.7 +/- 9.4% of PBMC from IgA nephritic patients, patients with chronic glomerulonephritis, and controls respectively expressed IL-2R (p greater than 0.05). However, the individual T-cell subsets bearing IL-2R were distinctly different between the IgA nephritic patients and patients with chronic glomerulonephritis or healthy controls. IgA nephritic patients had increased activated CD4+ lymphocytes (with IL-2R) (p less than 0.025) and reduced activated CD8+ lymphocytes (p less than 0.025). Our study suggests a defective immunoregulation in IgA nephropathy with enhanced T-helper/inducer and reduced T-suppressor/cytotoxic activity when stimulated with mitogen and probably, during clinical exacerbation. PMID- 2977311 TI - Germanium-induced nephropathy: report of two cases and review of the literature. AB - We report two cases of renal failure following long-term ingestion of germanium dioxide (GeO2) and comment on eight other cases reported in Japan. Ge-induced nephropathy is characterized by insidious onset of renal failure without proteinuria or hematuria after oral intake of Ge-containing compounds for more than several months, and by degeneration of renal tubular cells with minor glomerular abnormality in histology. When patients ceased to ingest Ge compounds, renal function gradually recovered but never returned to the normal range. Serious extrarenal complication can contribute to an unfavorable prognosis. PMID- 2977312 TI - Peripheral angioplasty--10 years on. PMID- 2977314 TI - Finding and dilating renal artery stenosis for hypertension. PMID- 2977313 TI - Radiological screening for atlanto-axial instability in Down's syndrome. AB - Two hundred and twenty Down's sufferers of all ages in the Sheffield district had their cervical spine X-rayed as part of a screening programme to ascertain atlanto-axial instability. Fifteen patients (6.8%) had an abnormal predental distance with four (1.8%) showing atlanto-axial subluxation. Seventy per cent of abnormal findings were in the under-25 age group. Only one child had clinical manifestations. Patients under the age of 25 years are most likely to participate in sports and are thought to be the age group most at risk. Degenerative changes in the cervical spine were found at an early age in patients with Down's syndrome, and there is both an increased incidence and severity of degenerative change at higher levels with increasing age. PMID- 2977315 TI - [Limitations of spontaneous reporting and ways of improving the quality of the data]. PMID- 2977316 TI - [Use of gold salts in Still's disease]. PMID- 2977317 TI - [Remarks on incidence of infections observed during 4 years of ambulatory practice]. PMID- 2977318 TI - [Aneurysm of the left ventricle. Natural history and course after surgery in adult and aged patients. Review of the literature and personal experience]. PMID- 2977319 TI - [Chemotherapy of advanced ovarian carcinoma. I. First-choice treatment]. PMID- 2977320 TI - [A rare manifestation of Hodgkin's disease: splenic abscess]. PMID- 2977321 TI - [Diet therapy of renal insufficiency]. PMID- 2977322 TI - Effect of gangliosides on primary and secondary plaque-forming cells responses and on suppressor T-cells induction. AB - This study was designed to determine the suppressive effect of bovine brain gangliosides on primary and secondary anti-sheep red blood cell (SRBC) plaque forming cells (PFC) response. Spleen cells from inbred C57BL/6J mice were used as sources of lymphocytes. The result showed that levels of gangliosides at 50 and 100 micrograms/ml suppressed primary anti-SRBC PFC responses about 30% and 60%, respectively, of normal control. In contrast, secondary anti-SRBC PFC were not suppressed by the same levels of gangliosides. Likewise, PFC response from mixing SRBC-primed T-cells with naive B-cells was not suppressed by gangliosides. In addition, induction of suppressor T-cells was reduced by 50 and 100 micrograms/ml of gangliosides, 57% and 63%, respectively, versus 86% of suppression of PFC response by the control suppressor T-cells. These results further suggest the role of gangliosides as immunomodulators. PMID- 2977323 TI - [Choice of candidates and perioperative management in coronary artery bypass graft]. PMID- 2977324 TI - The toxicity and mutagenicity of S-3-iodo-N-(1-ethyl-2-pyrrolidinyl)methyl-2 hydroxy-6-methoxybenzamide (IBZM), a new CNS D-2 dopamine receptor imaging agent. AB - IBZM is one of several benzamide derivatives showing a high affinity for the CNS D-2 dopamine receptor. Carrier-free [123I]IBZM is potentially useful as a nuclear medicine imaging agent for investigating CNS D-2 dopamine receptor in humans. This study describes the acute toxicity of IBZM in the rat and rabbit, its subchronic toxicity in the rabbit and its mutagenicity measured by the Ames test. IBZM had a 24 hour LD50 of 400 mg/kg in the rat and 50 mg/kg in the rabbit when administered i.v. Deaths occurred within minutes of dosing. Some necrosis was evident at the injection site in IBZM treated animals which was not found in the controls. No gross or histological differences between experimental animals and controls were evident in surviving animals when necropsied 14 days after dosing. Repeated exposure of rabbits to IBZM at a total cumulative dose of 100,000 times the expected clinical dose revealed no consistent changes in hematology, blood chemistry, blood enzymes or tissue pathology. IBZM was not mutagenic in the modified Ames assay with or without metabolic activation in the TA98 and TA100 tester strains. It is therefore unlikely that acute adverse effects will be associated with the diagnostic use of this drug in man. PMID- 2977325 TI - Beta-lipotropin-like immunoreactivity in intraglandular colloid from pituitary intermediate lobe cells. AB - The aim of this paper is the identification of beta-lipotropin (beta/LPH) as a peptide present in intraglandular colloid (the holocrine secretion of cells in the marginal half of the bovine pituitary intermediate lobe). beta/LPH, although not an opioid peptide itself, contains the peptide beta-endorphin. The methodology used allowed detection of beta/LPH when present in the samples in sufficient amounts. PMID- 2977326 TI - The hypertensive heart: pathogenesis, variants, and prognostic value. AB - We examine the heart's involvement in arterial hypertension, reporting on several studies of hypertensive patients showing that left ventricular myocardial mass is a significant prognostic indicator of essential hypertension and that left ventricular hypertrophy (LVH) correlates with an induced increase in Ca2+ concentration in platelets. We also consider the LVH variant of asymmetric hypertrophy and the role of the hypertensive heart in coronary insufficiency, and we speculate on the significance of the degree of LVH and asymmetric hypertrophy as risk factors for predicting cardiac complications of essential hypertension. PMID- 2977327 TI - Primary hypertension: left ventricular mass and function, sympathetic nervous system activity, and therapy. AB - Left ventricular (LV) mass is a predictor of morbidity in patients with hypertension. To elucidate the mechanisms of left ventricular hypertrophy (LVH) in primary hypertension, we examined the relationships of LV mass, arterial blood pressure, and plasma norepinephrine--as a marker of sympathetic nervous system tone--in three populations of patients with mild or moderate hypertension. We found linking of neural tone with (a) arterial blood pressure in mild and moderate hypertension, (b) LV mass in moderate hypertension, and (c) diastolic function in mild hypertension, suggesting that neural tone is a factor in both the pathogenesis and the sequelae of hypertension. Additionally, elevated norepinephrine level may reflect subtle loss of diastolic function before hypertrophy is manifest. Both nondrug and drug therapy may regress LV mass after blood pressure reduction with disparate effects on neural tone. PMID- 2977328 TI - Autogenic training in the treatment and secondary prevention of essential hypertension: five-year follow-up. AB - Ninety patients with essential hypertension were followed for 5 years. Initially the patients were randomized into two groups: (a) an experimental group consisting of 44 patients who received autogenic training and (b) a control group of 46 patients who did not receive any behavioral intervention. By the end of the follow-up period, the experimental group was significantly different from the control group, with reduced blood pressure (by 5.8 mm Hg systolic and 3.2 mm Hg diastolic vs. 4.3 mm Hg systolic and 2.0 mm Hg diastolic), a smaller increase in left-ventricular myocardial mass (14.6 g vs. 38.2 g), improved psychological indices, and a decrease in the number of sick days of leave. Autogenic training appeared to be more effective in patients with mild hypertension than in those with moderate hypertension and the results were comparable with those obtained with regular medication. PMID- 2977329 TI - Echocardiography, hypertension, and left ventricular mass. AB - The greater sensitivity of echocardiography than electrocardiography has revealed left ventricular hypertrophy (LVH) to be present in 20% or more of patients with systemic hypertension, with the exact prevalence dependent both on how a population is selected and on the sex, race, and possibly age composition of its members. LVH is more closely related to blood pressure recorded in the patient's natural setting during normal activity or exercise-whether measured by portable recorder or home manometer-than to blood pressure measured by the physician. A subgroup of patients with mild essential hypertension exhibit high cardiac output and evidence of supernormal myocardial contractility in the absence of LVH, whereas among patients with more sustained hypertension, high ejection fractions may be associated with marked concentric LVH. Recent data indicate that echocardiographic detection of LVH identifies mildly hypertensive patients at significant risk, a finding that may aid identification of patients for drug treatment. PMID- 2977330 TI - Isoenzyme patterns as a tool in taxonomy of chlorococcal algae. AB - From isolated lichen algae of the genus Trebouxia, De Puymaly isoenzymes of phosphoglucomutase, malate dehydrogenase and superoxide dismutase were studied by isoelectric focusing in hybrid immobilized pH gradients. Phosphoglucomutase has proven to be a suitable marker since it is highly variable whereas malate dehydrogenase and superoxide dismutase are less variable. The focusing patterns are species specific and provide a basis for a reevaluation of species classification of Trebouxia. PMID- 2977333 TI - The influence of the type of occupation on return to work after myocardial infarction, coronary angioplasty and coronary bypass surgery. AB - Between January 1980 and December 1983 the medical and social status of 423 patients who were considered candidates for aortocoronary bypass surgery (ACBS) was assessed by a questionnaire, at a mean of 16 months after coronary angiography. Of these patients 54 had refused surgery, 15 were re-operated, 23 had angioplasty and seven had died on the waiting list. After exclusion of these 117 patients, 306 remained, who form the basis of this report. Fifty three patients (17%) had retired before surgery, four (1.3%) had died perioperatively and 19 were on sick-leave for less than three months. Of those who were still employed pre-operatively, 102 (44.3%) went back to work, 85 (37%) had retired and 42 (18%) were on sick-leave for longer than three months. Significant differences were noted between the 102 working and the 85 retired patients as far as medical and social factors are concerned. Of the medical factors, post-operative freedom of symptoms (P less than 0.0001), postoperative exercise tolerance (P less than 0.0001) and completeness of revascularization (P less than 0.05) seemed to have influence on return to work. Of the social factors, age (P less than 0.0001), type of occupation (P less than 0.0002), duration of preoperative absence from work (P less than 0.001) and heavy manual work (P less than 0.05) showed significant differences between the groups. Since duration of preoperative absence from work is the only preoperative factor that can be modified, strategies for improving the return-to-work rate should aim at the shortening of waiting times for coronary angiography and ACBS. PMID- 2977331 TI - Electron microscopy and structural model of human fibronectin receptor. AB - Highly-purified human fibronectin receptor (a heterodimer of two distinct subunits, alpha and beta) was studied using electron microscopy and a variety of preparative procedures. It was found that the receptor consists of a globular head approximately 80 by 120 A and two tails about 20 A thick and 180-200 A long. The whole complex is approximately 280 A long. At low concentrations of detergent the receptor forms doublets, triplets or rosettes associated with the tails which possess the transmembrane portion of the molecule. Computer-assisted structure prediction using the published amino acid sequence of both subunits showed differences in the secondary structure of the tails, the alpha-tail being rich in beta-strands, the beta-tail having five cysteine-rich repeats analogous to the EGF-like repeats of laminin. Estimates of the length of the tails from the predicted structure conformed well with the dimensions obtained from electron micrographs. PMID- 2977332 TI - Possible orientational constraints determine secretory signals induced by aggregation of IgE receptors on mast cells. AB - Three biologically active monoclonal antibodies (mAbs) specific for the monovalent, high-affinity membrane receptor for IgE (Fc epsilon R) were employed in analysing the secretory response of mast cells of the RBL-2H3 line to crosslinking of their Fc epsilon R. All three mAbs (designated F4, H10 and J17) compete with each other and with IgE for binding to the Fc epsilon R. Their stoichiometry of binding is 1 Fab:1 Fc epsilon R, hence, the intact mAbs can aggregate the Fc epsilon Rs to dimers only. Since all three mAbs induce secretion, we conclude that Fc epsilon R dimers constitute a sufficient 'signal element' for secretion of mediators for RBL-2H3 cells. The secretory dose response of the cells to these three mAbs are, however, markedly different: F4 caused rather high secretion, reaching almost 80% of the cells' content, while J17 and H10 induced release of only 30-40% mediators content. Both the intrinsic affinities and equilibrium constants for the receptor dimerization were derived from analysis of binding data of the Fab fragments and intact mAbs. These parameters were used to compute the extent of Fc epsilon R dimerization caused by each of the antibodies. However, the different secretory responses to the three mAbs could not be rationalized simply in terms of the extent of Fc epsilon R dimerization which they produce. This suggests that it is not only the number of crosslinked Fc epsilon Rs which determines the magnitude of secretion-causing signal, but rather other constraints imposed by each individual mAb are also important.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977334 TI - Outcome from the questionnaire adopted for the consensus conference. PMID- 2977335 TI - Psychological factors and return to work. AB - Return to work is considered to be a criterion of a good outcome after a myocardial infarction or coronary revascularisation procedure. In general, return to work is not necessarily correlated with cardiac state; a variety of psychological, social and economic factors influence whether or not the individual will return to gainful employment. Psychological variables to be considered include the reactions of patient and family, their understanding of the illness, the personality and behavioural characteristics of the patient, expectations of the results of treatment and attitudes towards disability. These will be influenced by aspects of the job itself or the working environment as perceived by the patient. Physicians' attitudes are strong factors in influencing the final decision on whether or not the patient goes back to work. Employers and fellow employees, by their attitudes to the illness, will also influence the decisions. It follows that assessment and management of psychological problems must be part of total patient care after a coronary event if the optimal number of patients are to return to work, confidence of their ability to cope with its demands. Though the actual way in which assessment is made and appropriate treatment undertaken may vary considerably, a study of available knowledge enables general guidelines on these aspects to be given. PMID- 2977336 TI - Vasorelaxant effects of atriopeptins in human internal mammary artery. AB - The vasorelaxant effects of the atriopeptins, AP-(103-126) and AP-(103-123) were compared in human internal mammary artery (IMA) rings which had been contracted with norepinephrine. AP-(103-126) completely relaxed the IMA with an IC50 within the physiologically relevant range (15 nM). However, relaxation with AP-(103-123) was greatly reduced compared to AP-(103-126), with complete relaxation observed in only half of the vessels tested and only at concentrations of 10-50 microM. These results indicate that the atriopeptin (AP) receptor in human arteries may be more similar to the AP receptor in rabbit rather than rat arteries. PMID- 2977337 TI - The thromboxane receptor antagonist, daltroban, protects the myocardium from ischaemic injury resulting in suppression of leukocytosis. AB - The cardioprotective action of the new selective inhibitor of thromboxane receptors, daltroban (BM 13.505), was studied in cats subjected to 3 h of coronary artery ligation followed by 2 h of reperfusion. In comparison with vehicle (physiological saline)-treated cats, daltroban (20 mg/kg per h i.v.) reduced the ischaemia-induced rise in the ST segment and prevented the development of a Q-wave in the ECG during reperfusion. This was paralleled by a significantly improved preservation of creatine-phosphokinase activity in the ischaemic myocardium. Daltroban significantly attenuated platelet ATP secretion, the U-46.619-induced contraction of the cat thoracic aorta ex vivo and the myeloperoxidase-associated generation of reactive oxygen species ex vivo. These effects could be largely attributed to the inhibition of ischaemia-induced leukocytosis. It is concluded that daltroban protects the myocardium from ischaemic injury and that this effect involves prevention of ischaemia-induced leukocytosis. PMID- 2977338 TI - [Dynamic characteristics of the dependence of the venous return on changes in the volume of the circulating blood]. AB - Changes of the venous return of blood to the right heart were studied in respect to changes in the total blood volume. The latters were coarsed by the automatical transfusions of blood, controlled by the feed-back loop. Dynamic components of the dependence under study were treated by the spectral analysis technique. The contribution of them to the mechanism of the shift in systemic circulation can prevail over known static approximations of this dependence. PMID- 2977339 TI - Ninety-eight fixed drug eruptions with provocation tests. AB - Ninety-eight patients having fixed drug eruptions were subjected to provocation tests. Cutaneous involvement was seen in 43, mucous membrane in 33 patients, while cutaneous and mucous membrane involvement was seen in 22 patients. The causative drugs were found to be trimethoprim-sulphamethoxazole (45), acetylsalicylic acid (24), hyoscine butylbromide (Buscopan) (8), ibuprofen (Brufen) (6), oxyphenbutazone (6), tetracycline hydrochloride (2), phenolphthalein (1) and phenobarbitone (1). There was evidence of cross sensitivity between oxyphenbutazone and phenylbutazone. PMID- 2977340 TI - Generalized eruptive pustular drug rash due to cephalexin. AB - A 38-year-old woman developed a generalized pustular eruption 18 h after taking an oral cephalosporin (cephalexin). After having excluded other possible pustular dermatoses, we concluded that our patient's eruption was drug-induced. PMID- 2977341 TI - Heparan sulfate proteoglycan from human and equine glomeruli and tubules. AB - 1. Proteoglycans were isolated from human and equine glomeruli or tubules by guanidine extraction and anion exchange chromatography. 2. These proteoglycan preparations contained about equal amounts of heparan sulfate and chondroitin sulfates. 3. During the preparation of glomerular or tubular basement membranes the main part of proteoglycans (greater than 50%) was extracted in the salt extract. Chondroitin sulfate proteoglycan was mainly found in the water and salt extracts of glomeruli and tubules, heparan sulfate proteoglycan in the deoxycholate extracts and the basement membranes. 4. The glomerular basement membrane (GBM) contains about 12% (human) or 20% (equine) of the proteoglycans of the total glomerulus. They consist of greater than 70% (equine) or 80% (human) of heparan sulfate. 5. Heparan sulfate proteoglycan was isolated from the proteoglycan preparations of human or equine glomeruli and tubules by additional treatment with nucleases and chondroitinase ABC followed by CsCl gradient centrifugation. 6. Protein accounts for about 40% (dry weight) of the heparan sulfate proteoglycans. Their amino acid composition is characterized by a high content of glycine, but 3-hydroxyproline, 4-hydroxyproline and hydroxylysine are lacking. 7. The biochemical characteristics of the heparan sulfate proteoglycan of human or equine glomeruli or tubules differ from that isolated from rat glomeruli by their higher protein content and their amino acid composition. The significance of these differences is discussed. PMID- 2977342 TI - Hematoporphyrin ethers--III. Cellular uptake and photosensitizing properties. AB - 1. The cellular uptake and the efficiency in sensitizing cells to photoinactivation were determined for hematoporphyrin (Hp) diphenyl ether, Hp dicyclohexyl ether and Hp dihexyl ether. 2. The phenyl diether was taken up by the cells to the same degree as was the clinically used porphyrin preparation photofrin II, while the dihexyl and notably the dicyclohexyl ether were taken up 3-4 times better. 3. Furthermore, the quantum yields for photoinactivation of cells were similar for the three diethers and twice as large as that for photofrin II. 4. Fluorescence- and absorption spectroscopy indicate that these findings are related to the fact that photofrin II is much more aggregated in the cells than are the three Hp diethers. 5. When cells loaded with the porphyrins are incubated with porphyrin-free medium containing serum a certain percentage of the cell-bound drug is removed: 14% for photofrin II, 28% for Hp diphenyl ether, 50% for Hp dicyclohexyl ether and 20% for Hp dihexyl ether. 6. With respect to cell uptake and retention of the dyes, the data did not show any uniform relationship to the polarity of the drugs, in contrast to what has been found earlier for Hp diethers of linear hydrocarbons. PMID- 2977344 TI - [Dermatophytosis]. PMID- 2977343 TI - Ouabain-insensitive Na-ATPase activity in the basolateral membrane from rat jejunum. AB - 1. In the basolateral membrane preparation of the rat enterocyte (jejunal tract) there is not only the well-known (Na,K)-ATPase activity, but also a ouabain insensitive Na-ATPase. 2. The Na-ATPase is not activated by anions or other monovalent cations. As a substrate, ATP cannot be replaced by other nucleotides. 3. The Na-ATPase is insensitive to ouabain and bumetanide, inhibited partially by furosemide and totally by ethacrynate. 4. The activation of Na-ATPase at different Na concentrations shows an hyperbolic curve (Km = 15.7 +/- 2.3 mM and Vmax = 204 +/- 19 nmoles Pi/mg protein per min) different from the sigmoidal curve (Km = 9.8 +/- 1.2 mM and Vmax = 640 +/- 15 nmoles Pi/mg protein per min) shown by (Na,K)-ATPase. 5. These results are compared with the corresponding ones found in other animals and tissues in which the Na-ATPase was found. 6. The Na ATPase activity can be interpreted as the enzymatic correspondent of a ouabain insensitive Na pump, present in the basolateral membrane of the enterocyte different in behaviour with respect to the known Na pump. PMID- 2977345 TI - [Behavior of plasma beta-endorphin and treatment with non-steroidal anti inflammatory agents (NSAID)]. PMID- 2977346 TI - [Echocardiographic analysis of anatomical and functional changes in the left heart ventricle during antihypertensive treatment with nicardipine]. AB - Changes in hemodynamic variables regulating systolic function were assessed by M mode echocardiography, under 2D control, in 11 patients with primary uncomplicated hypertension treated with nicardipine (60 mg/daily). At the end of treatment (8 weeks) blood pressure and end-systolic stress were greatly reduced (p less than 0.001), and systolic fractional shortening was increase (p less than 0.02). The percentage increase in fractional shortening was correlated with a decreased in end-systolic stress (r = 0.67). The index of left ventricular performance (systolic pressure/end-systolic dimension ratio) was reduced: no variation was found in the hypertrophy-independent index of the inotropic state (systolic pressure/end-systolic dimension ratio normalized for posterior wall thickness). No change in the inotropic state was caused by a negligible sympathetic reflex outflow: heart rate also remained unchanged. The only index of left ventricular hypertrophy which was slightly reduced after two months of treatment was left ventricular mass (p less than 0.001). In conclusion, the short term treatment with nicardipine, in hypertensive patients, showed powerful effects in the improvement of systolic function; the possible regression of left ventricular hypertrophy must be confirmed in long-term therapy. PMID- 2977347 TI - [Atrial involvement in arrhythmogenic right ventricular dysplasia: primary or secondary? Description of a case of occult ventricular dysplasia with right atrial enlargement and exclusively sinoatrial arrhythmia]. AB - The atrial involvement in patients with arrhythmogenic right ventricular dysplasia is very rarely described. We here describe a patient with right atrial enlargement suffering from classical sinus and atrial electrical disease (tachycardia-bradycardia syndrome); the associated echo-angiographic aspects of the right ventricle are compatible, even in the absence of ventricular tachyarrhythmias, with a concealed form of right ventricular dysplasia. A common pathogenetic mechanism for the atrial and ventricular involvement is hypothetically considered. PMID- 2977348 TI - [Angioplasty in patients with multivessel coronary disease. Indication criteria and short- and medium-term results]. PMID- 2977349 TI - [Percutaneous coronary angioplasty in evolving myocardial infarction]. PMID- 2977350 TI - [Complications of percutaneous coronary angioplasty and significance of surgical stand-by]. PMID- 2977352 TI - [Percutaneous coronary angioplasty in Italy: data of the Italian Registry]. PMID- 2977351 TI - [Long-term results of coronary angioplasty: importance of restenosis]. PMID- 2977353 TI - Coliphage lambda to terminator lowers the stability of messenger RNA in Escherichia coli hosts. AB - The effects of the transcription terminators to and tfd on the overall high-level expression of a human interferon-beta gene (IFN-beta) in Escherichia coli hosts were compared. Deletion mapping shows that mRNA lability is caused by sequences at or near the lambda terminator to stem-loop structure. Extensive RNA secondary structure in this region indicates a potential RNase III cleavage/binding site. In RNase III- E. coli hosts, IFN-beta synthesis is indeed considerably enhanced. The bacteriophage tfd terminator does not confer this mRNA labilization phenomenon. In all cases, RNA level and stability correlate with the level of IFN beta synthesized in the cell. In the system described, ongoing translation stabilizes mRNA only moderately. PMID- 2977354 TI - Retroregulation of an int-lacZ gene fusion in a plasmid system. PMID- 2977355 TI - Structural analysis of mouse S-antigen. AB - Mouse S-antigen clones were isolated from a mouse retinal cDNA library using a bovine S-antigen cDNA probe. The largest clone (MSC-242) comprised 1532 bp and contained the entire coding sequence. The nucleotide sequence homology between the mouse and bovine coding regions was 84%, while non-coding regions appeared to be more divergent. The deduced amino acid sequence indicated that the mouse S antigen had 403 residues and its molecular ratio was 44,930. An overall amino acid sequence similarity of 84% was observed between the mouse and bovine proteins. This degree of similarity dropped to 60% and 47% at the N and the C termini, respectively. The local homology with alpha-transducin observed in the bovine proteins, including the putative phosphoryl and rhodopsin binding sites, was conserved in the mouse as well. There was no overall sequence similarity with other proteins listed in the National Biomedical Research Foundation (NBRF) protein sequence database. Among the uveitopathogenic sites for experimental autoimmune uveitis (EAU), peptides N and M were identical to their bovine counterparts. Peptides 3 and K, however, were more divergent. The short repeats within these peptides were conserved. PMID- 2977356 TI - Characterization of polyoma virus early proteins expressed from vaccinia virus recombinants. AB - We previously reported that live recombinant vaccinia viruses (VV) encoding either the large T (LT) or middle T (MT) antigens of polyoma virus (PyV) were able to induce rejection of tumors caused by PyV-transformed cells [Lathe et al., Nature 326 (1987) 878-880]. Here we present evidence that PyV early proteins expressed by the recombinants retain the biochemical characteristics of their authentic counterparts despite the cytopathic effect of VV infection. VV-encoded LT is a nuclear phosphoprotein, with specific DNA binding, ATPase and nucleotide binding activities. VV-expressed MT associates with cellular kinases, particularly with pp60c-src, by which it is phosphorylated in vitro. Expression levels of LT and MT reached 10(6) molecules per infected cell. The use of VV as a vector is encouraged by the high expression level obtained and because VV infection does not seem to prevent appropriate post-translational processing of proteins encoded by VV recombinants. PMID- 2977357 TI - Cloning and physical characterization of katE and katF required for catalase HPII expression in Escherichia coli. AB - Two genes, katE and katF, affecting the synthesis of catalase HPII in Escherichia coli, have been cloned. The multistep cloning protocol involved: screening for the tet gene in a transposon interrupting the genes, selecting DNA adjacent to the transposon, and using it to probe a library of wild-type DNA to select clones from which katE and katF were subcloned into pAT153. The clones were physically characterized and the presence of the genes confirmed by complementation of their respective mutations. The location of the transposon insertions in the two genes was determined by Southern blotting of genomic digests to further confirm the identity of the cloned genes. A 93-kDa protein, the same size as the subunit of HPII, was encoded by the katE plasmid, indicating that katE was the structural gene for HPII. A 44-kDa protein was encoded by the katF plasmid. PMID- 2977358 TI - Two human genes isolated by a novel method encode DNA-binding proteins containing a common region of homology. AB - Two cDNAs encoding new DNA-binding proteins (Dbps) have been cloned using a human placenta lambda gt11 recombinant cDNA library and DNA fragments as probes. Hybrid proteins expressed by the lambda gt11 cDNA library were blotted onto nitrocellulose filters, and incubated with three different radio-labeled DNA probes containing the human epidermal growth factor (EGF) receptor enhancer or the human c-erbB-2 promoter. Two kinds of clones, named dbpA and dbpB, showed high affinities for the DNA probes. The comparison of the nucleotide and the deduced amino acid (aa) sequences between these two cDNAs indicated that 100 of 109 aa located in the central region of these two Dbps were identical. The dbpA and dbpB-coded proteins also had an affinity for other cDNA probes such as the human c-ski gene, but not for poly(dI-dC).poly(dI-dC), suggesting that the sequence(s) recognized by the dbpA and dbpB-coded proteins may occur frequently, or that these proteins bind to DNA non-specifically in a different manner from that of histones. A simple method, described in this paper, can be used to isolate cDNA clones encoding Dbps. Strategies used for the detection of sequence specific and non-specific Dbps are discussed. PMID- 2977359 TI - [Hygienic evaluation of the conditions of the dairying process and the quality of milk products at subsidiary farms]. PMID- 2977360 TI - [Surface-active agents in the environment and human health]. PMID- 2977361 TI - [Various problems of the mathematical modeling of changes in the health status of the population in relation to atmospheric air pollution]. PMID- 2977362 TI - [Substantiation of hygienic standardization of ofunak in reservoir water]. PMID- 2977363 TI - [Hygienic evaluation of the air in animal housing]. PMID- 2977364 TI - [Oncohygienic evaluation of the possibility of using active sludge]. PMID- 2977365 TI - [A methodologic approach to the study of the physician's activities in health education]. PMID- 2977366 TI - [Hygienic problems of allocating and planning for milk pasteurization and processing plants at subsidiary farms]. PMID- 2977367 TI - [Standardization of the aluminum level in drinking water of various degrees of hardness]. PMID- 2977368 TI - [Effect of the school curriculum at a pulmonologic sanatorium on the functional status of children]. PMID- 2977369 TI - [Problems of occupational hygiene in the production of synthetic detergents (review of the literature)]. PMID- 2977370 TI - [Evaluation of the therapeutic and preventive effectiveness of the use of a polymer film in periodontal lesions in workers in the aluminum industry]. PMID- 2977371 TI - [Study of possible reproduction of causative agents of suppurative skin diseases in the lubricating-cooling liquid "Ukrinol-lM"]. PMID- 2977372 TI - [Effectiveness of introducing individual protection measures for workers in the phenazone industry]. PMID- 2977373 TI - [Characteristics of immunologic reactivity of workers at an epoxy resin plant]. PMID- 2977374 TI - [Comparative observation on atrial specific granules of secretive atriopeptides and secretive granules of beta-cells of pancreatic islets]. PMID- 2977375 TI - Characterization of the human HEXB gene encoding lysosomal beta-hexosaminidase. AB - The lysosomal enzyme beta-hexosaminidase A contains alpha- and beta-subunits that are encoded by the HEXA and HEXB genes, respectively. The human HEXB gene has been isolated and characterized. It is 45 kb long and is split into 14 exons. Of the 13 introns, 12 interrupt the coding sequences at homologous positions in the HEXA and HEXB genes. The 5' flanking region contains the functional HEXB gene promoter. While a fine-structure analysis has yet to be done, we note that the sequence is GC rich and has several GC boxes and one CAAT box. There are also sequences related or identical to a progesterone response element and an AP-1 binding motif. PMID- 2977376 TI - [Use of isotretinoin in severe polymorphous acne]. PMID- 2977378 TI - [Dermatologic aspects of HIV infections]. PMID- 2977377 TI - [Epidemiological data on infantile seborrheic dermatitis in 12 years of ambulatory care activity (1974-1985) at the Institute of Clinical Dermatology I and Pediatric Dermatology of the University of Milan]. PMID- 2977379 TI - A soluble 'anchorminus' interleukin 2 receptor suppresses in vitro interleukin 2 mediated immune responses. AB - The immunosuppressive effects of a recombinant soluble IL-2 receptor L chain (s IL-2R) were analyzed. S-IL-2R protein was obtained from the conditioned medium of L cells transfected with a mutant cDNA clone encoding the extracytoplasmic portion of the IL-2 receptor (IL-2R) and was purified to homogeneity by an IL-2 coupled sepharose column, following by reverse phase chromatography (HPLC). Soluble IL-2R protein thus prepared retained the ability to bind IL-2 specifically and suppressed the in vitro IL-2-mediated immune responses, including proliferation of IL-2-dependent cell line (CTLL-2), induction of secondary cytotoxic T lymphocytes (CTL) and the mixed lymphocyte reaction (MLR), but did not suppress the growth of IL-3-dependent cell line. Kinetic studies revealed that s-IL-2R exhibited the suppressive effects on the proliferative responses of alloantigen stimulated human tonsillar cells, only when added at an early stage, namely 0-48 h after culture onset, whereas cyclosporin A (CsA) exhibited an inhibitory effect only when added at between 0 and 24 h. This implies that s-IL-2R exerts its effect on an early stage of lymphocyte activation. The observed immunosuppressive effects of s-IL-2R suggest the possibility that s-IL-2R might be useful for the protection of rejection crisis in organ transplantation. PMID- 2977381 TI - Percutaneous balloon valvoplasty and angioplasty in children. PMID- 2977380 TI - Effect of cisplatin on release of lysozyme, plasminogen activator, leucine aminopeptidase and beta-hexosaminidase by murine peritoneal macrophages in vitro. PMID- 2977382 TI - Conservative treatment of abdominal stab wounds with omental protrusion. AB - Two cases of abdominal stab wounds with omental protrusion treated conservatively are presented. Omental protrusion in a patient without signs of shock or peritoneal irritation is not an absolute indication for explorative laparotomy. PMID- 2977383 TI - Pharmacokinetics of cyproterone acetate and its main metabolite 15 beta-hydroxy cyproterone acetate in young healthy women. AB - Cyproterone acetate (CPA) and 15 beta-hydroxy-cyproterone acetate (15 beta-OH CPA) have been quantitated in human plasma by a selective, automated HPLC assay in 8 young female subjects. Subjects received a single oral dose of 100 mg CPA and four weeks later, a single intramuscular dose of 300 mg CPA. Plasma levels in seven subjects could be evaluated pharmacokinetically. After intramuscular administration maximum CPA serum levels of 191 +/- 49 ng/ml were reached between 2 and 3 days. Postmaximal levels declined with a half-life of 4.3 +/- 0.7 days. After oral administration maximal serum levels of 255 +/- 110 ng/ml were measured between 2 and 3 h. CPA levels decayed biphasically with a terminal half-life of 3.6 +/- 1.3 days. Bioavailability after oral administration was nearly complete (88 +/- 20%). Metabolite levels were below CPA levels until 4 days (i.m.) and 6 h (p.o.) and exceeded CPA levels later on. 15 beta-OH-CPA levels declined with half lives similar to those of corresponding CPA levels, indicating that terminal half life of the metabolite in serum reflects its rate of formation rather than its rate of elimination. Areas under the metabolite serum level curves after intramuscular and oral administration were 10 to 15% higher than the corresponding areas for unchanged CPA. Since CPA and 15 beta-OH-CPA contribute to the antiandrogenetic efficacy there is good reason to assume that the intramuscular administration of CPA might be equally effective in the therapeutical use as the oral administration. PMID- 2977384 TI - An in-vitro evaluation of the cellular uptake and intraphagocytic bioactivity of clarithromycin (A-56268, TE-031), a new macrolide antimicrobial agent. AB - Erythromycin base and its 6-0-methyl derivative clarithromycin were actively accumulated 7.3 +/- 1.2-fold and 9.2 +/- 2-fold respectively by human neutrophils in vitro. The intraphagocytic bioactivities of the antimicrobial agents were investigated using the combination of a radioassay, colony counting method and a fluorescence microassay which facilitates the distinction between intracellular bacteriostatic and bactericidal mechanisms. Staphylococcus aureus, Listeria monocytogenes and Legionella micdadei were used as the test intraphagocytic microbial pathogens. Both agents were found to possess intracellular bioactivity for all three species of bacteria with clarithromycin being consistently more active than erythromycin. Under the assay conditions used both agents were bacteriostatic (intracellularly) for S. aureus and Leg. micdadei and bactericidal for List. monocytogenes. Clarithromycin is clearly a potent intraphagocytic antibiotic and potentially superior in this respect to erythromycin. PMID- 2977385 TI - Regulatory light chain-a myosin kinase (aMK) catalyzes phosphorylation of smooth muscle myosin heavy chains of scallop, Patinopecten yessoensis. AB - Regulatory light chain-a myosin kinase (aMK), which phosphorylates one of the myosin regulatory light chains, RLC-a, contained in the catch muscle of scallop, was also found to phosphorylate heavy chains of scallop myosin. After incubation of myosin isolated from the opaque portion of scallop smooth muscle (opaque myosin) with aMK in the presence of [gamma-32P]ATP, about 2 mol of 32P was incorporated per mol of the myosin. The radioactivity was mostly found in the heavy chain at 0.26 M KCl. The pH-activity curve and MgCl2 requirement for the heavy chain phosphorylation were similar to those for RLC-a phosphorylation. In contrast, the dependency of activity on KCl concentration was different from that for RLC-a. The heavy chain phosphorylation activity decreased with increase in KCl concentration up to 0.06 M, and then increased at concentrations over 0.06 M to a maximum at around 0.26 M KCl. This complicated profile probably reflects the solubility of myosin, and the phosphorylation site may be located in the rod portion insoluble at low KCl concentrations. Phosphorylation of heavy chain did not change the solubility of the opaque myosin molecule at all. The acto-opaque myosin ATPase activity in the presence of Ca2+ was found to be decreased to less than one-fourth by the heavy chain phosphorylation. PMID- 2977386 TI - Chemical synthesis and expression of copper metallothionein gene of Neurospora crassa. AB - The gene coding for the Neurospora crassa copper metallothionein (MT) was synthesized and inserted in the lacZ' gene of pUC18 plasmid to give the same translational reading frame as the latter gene. The MT-beta-galactosidase fused gene was expressed in Escherichia coli to produce a fused protein in which the amino and carboxy termini of MT are linked to the beta-galactosidase through methionine residues. An MT derivative containing an extra homoserine residue at the carboxy terminus was prepared by cyanogen bromide cleavage of the fused protein followed by a reverse-phase HPLC separation. The spectral features of the MT derivative and its copper complex were similar to those of the corresponding native MTs. PMID- 2977387 TI - Partial purification and properties of UDP-N-acetylmannosamine:N acetylglucosaminyl pyrophosphorylundecaprenol N-acetylmannosaminyltransferase from Bacillus subtilis. AB - An enzyme which catalyzes the conversion of GlcNAc-PP-undecaprenol into ManNAc(beta 1----4)GlcNAc-PP-undecaprenol, a key lipid intermediate in the de novo synthesis of various teichoic acids, was partially purified from the 20,000 x g supernatant fraction of Bacillus subtilis AHU 1035 cell homogenate. By means of ammonium sulfate precipitation, gel chromatography, and ion-exchange chromatography, the enzyme was purified about 70-fold, giving a preparation virtually free from substances obstructive to measurement of the N acetylmannosaminyltransferase reaction. The enzyme was shown to be specific to UDP-ManNAc. The Km value for UDP-ManNAc was 4.4 microM, and the optimum pH was 7.3. The enzyme required 10 mM MgCl2, 0.3 M KCl, 25% glycerol, and 0.1% Nonidet P 40 to function at full activity. PMID- 2977389 TI - Quantitative analysis of retinoids in biological fluids by high-performance liquid chromatography using column switching. II. Simultaneous determination of etretinate, acitretin and 13-cis-acitretin in plasma. AB - An automated gradient high-performance liquid chromatographic method for the determination of etretinate, acitretin and 13-cis-acitretin in plasma was developed, using a column-switching technique. After protein precipitation with ethanol, 0.5 ml of the supernatant was injected onto a precolumn (17 mm x 4.6 mm I.D.), filled with 37-53 microns C18 Corasil. Polar plasma components were washed out using 1% ammonium acetate and 1% acetic acid-acetonitrile (8:2, v/v); the retained retinoids were then transferred to the analytical column (125 mm x 4 mm I.D., filled with 5-microns ODS material) in the backflush mode, separated by gradient elution and detected at 360 nm by UV detection. The limit of quantification was 2 ng/ml and the inter-assay precision in the concentration range 20-1000 ng/ml was between 0.9 and 4.0% for all three compounds. To optimize the recovery for etretinate (greater than 60%), protein was precipitated from plasma with ethanol before injection, instead of direct injection of plasma samples, and a mobile phase containing 20% acetonitrile, instead of pure water or buffer, was used. PMID- 2977388 TI - Solubilization and properties of UDP-D-glucose:N-acetylglucosaminyl pyrophosphorylundecaprenol glucosyltransferase from Bacillus coagulans AHU 1366 membranes. AB - The glucosyltransferase which catalyzes the conversion of GlcNAc-PP-undecaprenol into Glc(beta 1----4)GlcNAc-PP-undecaprenol in the presence of UDP-glucose was solubilized from Bacillus coagulans AHU 1366 membranes by treatment with 0.1% Triton X-100 and partially purified by means of column chromatography on Sephacryl S-300 and DEAE-Sephacel. The final preparation was virtually free from other enzymes involved in the de novo synthesis of teichoic acid. The enzyme had a pH optimum of 6.6-8.0 and a Km value for UDP-glucose of 21 microM. The enzyme required 40 mM MgCl2, 0.6 M KCl, and 0.1% Nonidet P-40 for full activity. PMID- 2977390 TI - Development of a solid-phase extraction technique for alpha-human atrial natriuretic peptide in human plasma. AB - A reliable extraction method was developed for alpha-human atrial natriuretic peptide (alpha-hANP) using Bond Elut C8 columns in tandem. This involved activation of the columns using methanol followed by a water wash to remove the excess methanol. Plasma (1 ml) was then added and a vacuum applied until all was drawn through. Excess protein and other endogenous compounds were removed by washing the columns with water and elution of the alpha-hANP was achieved with 0.75 ml acetonitrile-water-trifluoroacetic acid (80:19.8:0.2, v/v/v). Samples were evaporated under nitrogen and reconstituted in radioimmunoassay buffer ready for analysis. The recovery of alpha-hANP from plasma using this method was found to be 90% +/- 0.6% [mean +/- standard error of the mean (S.E.M.); coefficient of variation (C.V.) = 1.5%] which will allow more precise measurement of the peptide than is presently available. With this high precision of analysis available, having a limit of detection of 0.4 fmol/ml and a range of 0 to 32 fmol/ml, a low dose infusion of alpha-hANP was conducted and the changes in plasma concentration were followed. PMID- 2977391 TI - Simultaneous determination of niacin and niacinamide in meats by high-performance liquid chromatography. PMID- 2977392 TI - Food increases the bioavailability of acitretin. AB - Eighteen healthy male volunteers received 50 mg oral doses of acitretin on two occasions, according to a random crossover design. Acitretin was administered during a complete fast or following a moderate breakfast. Plasma samples were obtained at various times and the concentration of acitretin and its 13-cis isomeric metabolite (Ro 13-7652) were quantified by a specific HPLC assay. The AUC0-15 for acitretin was increased when administered with food for all subjects (except one) with a mean increase of 90% (from 1175 to 2249 ng/ml.hr). The maximum plasma concentration of acitretin (Cmax) was increased by 70% when administered with food (from 245 to 416 ng/ml), while the time to reach Cmax was unaffected. The ratio of AUC of Ro 13-7652 to acitretin was the same for both the fasted and fed conditions; therefore, the formation of metabolite was not influenced by concomitant ingestion of food. The presence of food increases the apparent bioavailability of acitretin. A likely mechanism behind this observation is an increase in acitretin solubility in addition to an increase in the lymphatic absorption and a prolonged residence time of the drug in the gastrointestinal tract. PMID- 2977393 TI - Amlodipine pharmacokinetics in healthy volunteers. AB - In the present study we investigated the pharmacokinetics and comparative bioavailability of three oral doses of amlodipine in 12 healthy male volunteers. A randomized, open-label, three period crossover study design was employed. Each subject received, on three separate occasions a single oral dose of 2.5, 5 and 10 mg amlodipine. Standing diastolic blood pressure was reduced by 1.1, 4.8 and 8 mmHg six hours after 2.5, 5 and 10 mg amlodipine, respectively. There were no significant changes in pulse rate, nor on the EKG. The curves for the mean plasma concentrations versus time for the three doses showed parallel time-courses. Highly significant positive correlations were observed between dose and AUC (0-72 hrs) and between dose and Cmax. However, dose corrected AUC and Cmax were 10-20% lower with 2.5 mg, than with 5 and 10 mg. Peak levels were achieved 5.6 to 6.4 hours postdose. Half lives were 31.2, 33 and 36.8 hours for 2.5, 5 and 10 mg respectively. Headache was the most common side effect, and was more frequently observed with the highest dose. In summary, linear relationships were found between the dose and the plasma levels of amlodipine. Decreases in standing diastolic blood pressure were also dose related. Because of its long half-life and gradual absorption, amlodipine should be effective in lowering blood pressure given once daily and the incidence of side effects due to rapid absorption should be minimized. PMID- 2977394 TI - The effects of renal function on the disposition of isradipine. AB - The effect of renal function on isradipine kinetics was examined in four groups of subjects (N = 55) who had normal or impaired renal function. Each subject received isradipine orally as a 10-mg capsule. Serial blood samples were obtained from 0 to 48 hours postdose and the isradipine plasma concentrations determined by radioimmunoassay. Kinetic parameters, Cmax, lambda 3, t 1/2, AUC, CL'o (oral clearance), and CLo (oral clearance standardized to body weight) were determined. Marked intersubject variability of the pharmacokinetic parameters was observed. No statistically significant differences (P greater than .05) were found for AUC, Cl'o, and Clo parameters when renal impairment groups were compared with controls. AUC values were lower (P less than .05), however, for the group with severe renal function impairment than for groups with mild or moderate renal function impairment. No significant correlations (r = -.23, P greater than .05; and r = .13, P greater than .05, respectively) were found between creatinine clearance (CLCR) and CLo and between age and CLo. Considering the interpatient variability in isradipine disposition and the lack of significant differences in CLo between groups, no clear-cut dosing regimen alterations, based on single-dose data, are warranted in renal impairment. PMID- 2977395 TI - [Papilledema caused by minocycline: apropos of a case]. AB - Benign intracranial hypertension with papilloedema developed in a 18-year-old woman following Minocycline administration. Tetracycline therapy was prescribed for acne vulgaris. One month after the beginning of the treatment, she presented with headache, nausea and vomiting; there were no visual symptoms. Visual acuity and visual field were normal, fundus examination showed bilateral papilloedema. After Minocycline was discontinued and steroid therapy was administrated, symptoms rapidly resolved and papilloedema disappeared. Minocycline is known to penetrate into the central nervous system more effectively and to have a greater lipoid solubility than the other antibiotics of the same group. However the pathogenesis of benign intracranial hypertension after Minocycline therapy remains unknown. PMID- 2977396 TI - The clinical relevance of tumor marker CEA, CA 19-9 in regional chemotherapy of hepatic metastases of colorectal carcinoma. AB - Up to December 1986, 50 patients with documented hepatic metastases from colorectal carcinoma were treated with 5-fluoro-2-deoxyuridine (FUDR) using Infusaid pumps. The response of liver metastases to regional chemotherapy was studied by computerized tomography (CT) and carcino-embryonal antigen (CEA), and/or CA 19-9 antigen serum assays. Preoperative CEA values were pathological in 94% of the patients but only 48% had a pathological concentration of the antigen CA 19-9 of over 37 U/ml. The course of CEA and CA 19-9 in combination with the arterial angio-CT reflected the response of liver metastases to regional chemotherapy. A decrease or normalisation of CEA and CA 19-9 after the beginning of therapy is an indication of partial or complete remission of metastases (68% of the patients showed lowered CEA serum values). If the marker continues to rise in serum this is a danger signal of progression of liver metastases or of extrahepatic tumor spread if the tumor stage in the liver remains unchanged. PMID- 2977397 TI - Neuroimmunomodulation in cancer patients: correlations between melatonin and beta endorphin blood levels and T helper/suppressor ratio. AB - The pineal gland and opioid peptides play roles in the neuroendocrine control of immunity. Both neuroendocrine and immune dysfunctions have been observed in cancer but the importance of the altered secretion of neurohormones in the immunoincompetence of cancer patients has never been investigated. This study concomitantly evaluated neuroendocrine and immune functions in 40 patients with early or advanced neoplastic disease. In each patient, melatonin and beta endorphin blood levels and lymphocyte subtypes were determined on venous blood samples collected during the morning. Metastatic patients had lower melatonin levels and a lower T4/T8 ratio than patients without metastases but no significant correlation was found between melatonin and the T4/T8 ratio. beta endorphin levels appeared to be normal in all patients. These results suggest that melatonin and beta-endorphin secretion have no role in determining immune dysfunctions in cancer. PMID- 2977398 TI - Biomaterials and immune system: cellular reactivity towards PTFE and Dacron vascular substitutes pointed out by the leukocyte adherence inhibition (LAI) test. AB - Thirty-eight patients affected by peripheral vascular insufficiency, and twelve healthy volunteers, were submitted to a cellular immunity test: LAI test, in which leukocytes fail to adhere to glass in contact with a sensitizing antigen. Patients were divided as follows: Class 1: Dacron+PTFE grafted patients, Class 2: Dacron, Class 3: PTFE, Class 4: controls. Total leukocytes, mononuclear cells, T and B lymphocytes were used as cellular populations. Finely minced Dacron and PTFE fabric vascular prostheses were employed as targets. This research showed that a T cellular immune-reactivity towards Dacron and PTFE respectively occurs in Dacron and PTFE grafted patients, and that reactivity is greater in case of Dacron. Wider researches are required to state the immune system role in fabric prostheses patency; at this regard, must be kept in mind that T lymphocytes release thrombogenic factors in course of cellular immune response. PMID- 2977399 TI - Collagen arthritis in rats: the importance of humoral immunity in the initiation of the disease and perpetuation of the disease by suppressor T cells. AB - Arthritis could be passively transferred with a serum concentrate from collagen arthritic rats to nude rats and cyclosporin-treated, type II collagen-tolerant rats. These findings suggest that collagen arthritis could be inducible by humoral immunity alone in the absence of cellular immunity to type II collagen or functional T cells. In addition, passive arthritis induced by anticollagen antibody is a mild, transient disease from which the animals normally recover and the rats that have recovered from passive arthritis are resistant to develop a second phase of arthritis following a second administration of anticollagen antibody or the subsequent challenge with type II collagen. However, when a serum concentrate was transferred while cyclosporin was administered continuously, transferred arthritis in cyclosporin-treated, type II collagen-tolerant rats lasted as long as cyclosporin treatment and arthritis was significantly enhanced compared to those of naive recipients. Further, enhancement and prolongation of passively transferred arthritis in nude rats was observed. Furthermore, treatment with cyclophosphamide reversed acquired resistance to collagen arthritis subsequent to recovery from passive arthritis. These findings suggest that suppressor T cells might, at least in part, affect the clinical course of collagen arthritis and reverse acquired resistance to arthritis. PMID- 2977400 TI - Antigen specific down regulation of murine collagen induced arthritis: T suppressor cell circuits in arthritis immunotherapy. AB - The present article summarizes a series of experiments which have been performed to describe an antigen-specific suppressor cell pathway for the suppression of the erythema and edema associated with an animal model of rheumatoid arthritis, collagen induced arthritis (CIA). Initial studies utilized the adoptive transfer of splenic cell subpopulations to establish the presence of suppressor cells in lymphoid tissues of mice which were suppressed for collagen induced arthritis. Subsequent studies generated T cell hybridomas from animals which had been suppressed for collagen induced arthritis by a single injection of a large quantity of Type II collagen. The T cell hybridomas varied in their self surface expression of glycoproteins which are associated with genetically determined functions. The suppressor T cells generated, described a regulatory suppressor cell pathway comprised of at least afferent suppressor T cells and effector suppressor T cells. The cells act in an antigen-specific fashion with regard to the suppression of collagen induced arthritis but appear to be polymorphic in their recognition of the interstitial collagens. The studies, taken together, indicate that the use of antigen specific T suppressor cells in the form of T cell hybridomas can be utilized as a form of immunotherapy in experimental arthritis. PMID- 2977401 TI - AIDS in the workplace. Handicap discrimination laws and related statutes. PMID- 2977402 TI - What makes a good photosensitizer for photodynamic therapy? PMID- 2977403 TI - Biological action of colorless and yellow solutions of gold sodium thiomalate on thrombin activity and the mixed lymphocyte reaction. AB - Gold sodium thiomalate is a pale yellow powder which forms a colorless solution when added to sterile water. The marketed form of gold sodium thiomalate is a pale yellow solution. The yellow color develops as a result of the sterilization process. This study demonstrates that the physical change induced in the drug by the sterilization process has no effect on the action of gold sodium thiomalate on the serine esterase thrombin, nor on the inhibition of the mixed lymphocyte response. Thus it is unlikely that the yellow component is responsible for benefit in rheumatoid arthritis. If the components creating the yellow color cause toxicity, the preparation and/or formulation of the drug should be changed. PMID- 2977404 TI - Captopril reduces the renal response to intravenous atrial natriuretic peptide in normotensives. AB - The interaction between atrial natriuretic peptide (ANP) and the renin angiotensin-aldosterone (RAA) system was studied in 6 healthy volunteers using the angiotensin-converting-enzyme (ACE) inhibitor, captopril. Each volunteer received, on separate occasions in random order, 48 hours treatment with a) low dose captopril (6.25 mg twice daily), b) 'high-dose' captopril (25 mg twice daily) and c) placebo prior to an infusion of synthetic human ANP (99-126). Resting plasma ANP levels were significantly (P less than 0.01) higher on treatment with 'high-dose' captopril when compared with low-dose captopril or placebo. 'High-dose' captopril reduced mean arterial blood pressure (BP) and significantly (P less than 0.01) reduced the natriuretic response to the human ANP infusion. These results support the hypothesis that ACE is involved in ANP metabolism. The reduced renal response to ANP during treatment with captopril may reflect the dependence of ANP on adequate renal perfusion pressure and angiotensin II levels to exert its natriuretic effect. PMID- 2977405 TI - Platelet-activating factor and anti-platelet-aggregating factor in acute reduction of blood pressure following percutaneous transluminal renal angioplasty in patients with renovascular hypertension. AB - The mechanism of the acute fall of BP following percutaneous transluminal renal angioplasty (PTRA) was studied in four patients with renovascular hypertension caused by fibromuscular dysplasia. One hour after PTRA, systemic blood pressure and plasma renin activity in the ipsilateral renal venous blood decreased significantly (P less than 0.05), but the plasma noradrenaline level in ipsilateral renal venous blood increased significantly (P less than 0.05). At the same time, a platelet-activating factor (PAF) and an unidentified factor that inhibited the aggregation of rabbit platelets induced by PAF, arachidonic acid or ADP were detected in the ipsilateral renal venous blood, but were not found in the contralateral renal venous blood. Plasma noradrenaline level in cubital venous blood decreased significantly (P less than 0.05) after 24 hours as compared with that before PTRA and BP also maintained the normal level. These results suggest that the reduction in plasma renin activity is associated with the acute reduction in BP following PTRA. PAF and an unidentified factor blocking the aggregation of platelets may be involved in ipsilateral renal venous blood following PTRA in patients with renovascular hypertension. The reduction in plasma noradrenaline level is an additional mechanism involved in maintaining normal BP following PTRA in the late stage. PMID- 2977406 TI - Amlodipine and captopril in moderate-severe essential hypertension. AB - The therapeutic usefulness of adding once-daily amlodipine (10 mg) for four weeks in moderate-severe hypertensive patients uncontrolled on low dose captopril (25 mg twice daily) alone was studied in 29 patients in a double-blind, placebo controlled two-way crossover comparison. Once daily amlodipine was shown to be an effective antihypertensive drug when combined with captopril. The amlodipine minus placebo differences in mean changes from captopril baseline values were: 18/-12 mmHg and -20/-12 mmHg for supine and standing systolic/diastolic pressures (P less than 0.001 for all four pressure variables). The combination was well tolerated, and no patient discontinued therapy. Five patients experienced ankle oedema and four patients reported flushing while receiving amlodipine/captopril. PMID- 2977407 TI - Effects of heptaminol AMP amidate on suppressor and helper function of murine T cells. AB - Heptaminol AMP amidate (HAA), a newly developed nucleotide derivative, was found to restore the immunosuppression in mice due to the induction of suppressor T (Ts) cells by concanavalin A (Con A) (50 micrograms/body). HAA also inhibited Con A-mediated in vitro induction of Ts cells. On the contrary, the administration of HAA in mice primed with keyhole lympet hemocyanin (KLH) (30 micrograms/body) caused an enhanced induction of antigen specific helper T (Th) cells. Effects of HAA on Ts and Th cells were found to be dependent on their level of induction. The administration of HAA also increased the spleen cell number and augmented the plaque forming cell response to some extent in cyclophosphamide treated mice. The present results suggested that HAA-mediated immunopotentiation was possible by a combined suppressive effect on Ts cells and enhancing effect on Th cells. PMID- 2977408 TI - [Review of morphologic changes in electrocardiograms of 70 patients with pheochromocytoma]. PMID- 2977409 TI - [Prognostic value of the electrocardiogram in dilated cardiomyopathy]. PMID- 2977410 TI - [Prevalence of ischemic heart disease among the rural population and its relation to dietary habits]. PMID- 2977411 TI - [Excessive body weight as one of the major risk factors of cardiovascular diseases in rural population]. PMID- 2977412 TI - [Evaluation of left-ventricular myocardial hypertrophy in patients with mitral valve insufficiency based on the data of electro- and echocardiography]. PMID- 2977413 TI - [Vladimir Kharitonovich Vasilenko]. PMID- 2977414 TI - [Non-invasive evaluation of left-ventricular function in hypertension]. AB - Left-ventricular functions were assessed by means of mechanocardiography in patients with essential hypertension in the absence of signs of heart failure. A significant prolongation of the isovolumic relaxation phase, rapid filling phase, and an increase in the amplitude of apex-cardiographic "a" wave were found in hypertensive patients showing no ventricular hypertrophy. Similar but more pronounced disorders were observed in patients with ventricular hypertrophy where, in addition, the pre-ejection period was prolonged and there were signs of asynergic contraction. The findings testify to diastolic dysfunction of the left ventricle as a result of its impaired compliance, already present in the early phases of essential hypertension. Mechanocardiography is a suitable method for the detection of cardiac dysfunction in essential hypertension. PMID- 2977415 TI - [Effect of compensatory myocardial hypertrophy on the electric stability of the heart]. AB - Dog heart was shown to be more prone to ventricular fibrillation at the stage of completed right-ventricular hypertrophy about 5 months after pulmonary arterial trunk was stenosed by 2/3 of the original lumen, as compared to normal heart. Electrical instability of the heart with completed right-ventricular hypertrophy increases in proportion to the magnitude of hypertrophy. PMID- 2977416 TI - [Hypertonic heart]. PMID- 2977417 TI - [Prevention of disorders of cardiac contractility with nicotinamide in adriblastin -related damage]. AB - Pretreatment with 20 mg/kg nicotinamide 3 days prior to a single intraperitoneal 6 mg/kg adriblastin injection, followed by repeated injections every second days for 1 week, prevented cardiac contractility disorders in adriblastin-treated rats, while their cardiac contractility was less prone to hypoxic depression and recovered better at subsequent reoxygenation. PMID- 2977419 TI - Distribution of zeranol in bovine tissues determined by selected ion monitoring capillary gas chromatography/mass spectrometry. AB - Bovine tissues, including liver, muscle, kidney, bile, serum, and urine, have been quantified by selected ion monitoring capillary gas chromatography/mass spectrometry to establish the distribution of the anabolic drug, zeranol, and its metabolites, taleranol and zearalanone, after administration of zeranol to 9 bovine animals. The method used to isolate, confirm, and quantify zeranol is undergoing validation by the United States Department of Agriculture, Food Safety Inspection Service (FSIS). Application of this method demonstrates utility in determining residue levels of zeranol in a variety of tissues with levels ranging over 4 orders of magnitude (i.e., 100 parts per trillion (ppt) to 1 part per million (ppm]. The analyte levels determined in this study complement previously reported pharmacokinetic data on the distribution of zeranol in addition to providing more specific information for taleranol and zearalanone. In this quantitative study it is shown that the liver is the main organ of deposition for zeranol, taleranol, and zearalanone, that taleranol is the main metabolite in the bovine, and that zeranol is efficiently eliminated. PMID- 2977418 TI - Metabolism of 3H-dehydroepiandrosterone sulphate by subjects with steroid sulphatase deficiency. AB - Patients with steroid sulphatase deficiency develop ichthyosis with accumulation of cholesterol sulphate in plasma and in the stratum corneum. The present study was undertaken to determine whether desulphation of the C19 steroid DHEAS is also impaired. The mean plasma concentrations of DHEA and androstenedione were significantly lower for patients than for controls (p less than 0.02 and 0.001) while the mean concentration of DHEAS was higher (p less than 0.002). Following intravenous administration of 3H-DHEAS, one patient failed to desulphate 3H-DHEAS as evidenced by an absence of urinary 3H-glucuronides. A second produced normal amounts of urinary 3H-glucuronides (indicative of desulphation capacity) in a baseline study but did not desulphate 3H-DHEAS following ampicillin treatment to alter gut microflora. A third patient had consistent sulphatase activity with and without ampicillin. PMID- 2977420 TI - [Lack of correlation between atrial natriuretic peptide and arterial pressure in hemodialysed patients]. PMID- 2977421 TI - Characterization of the interaction of human plasmin with its specific receptor on a group A streptococcus. AB - Certain Group A beta-hemolytic streptococci express a receptor that is capable of specifically binding the human plasma protease plasmin. Once bound, plasmin remains enzymatically active and is unregulated by its naturally occurring inhibitor alpha-2-antiplasmin (Lottenberg, R., C. C. Broder and M. D. P. Boyle, 1987. Infect. Immun. 55: 1914-1918). In this study certain characteristics of the interaction between plasmin and the receptor expressed on a group A beta hemolytic streptococcus, strain 64/14, were examined. Binding occurred optimally at physiologic pH and ionic strength. The KD was 5 x 10(-11) M and there were approximately 800 receptors per bacterium. Mouse passage of strain 64 had no significant effect on the KD of the receptor. Binding of plasmin to the bacteria was inhibited by lysine and epsilon-aminocaproic acid in a concentration dependent manner. Similarly these amino acids would displace pre-bound plasmin from the bacteria. These findings suggest a role for plasmin's high affinity lysine binding site in the interaction of plasmin with the bacteria. PMID- 2977422 TI - Phosphofructokinase activity in fibroblasts from patients with Alzheimer's disease and age- and sex-matched controls. AB - The activity of the enzyme phosphofructokinase (PFK) was comparable in cultured skin fibroblasts from eight patients with Alzheimer's disease and eight age- and sex-matched controls. Mean activities were similar in the two groups whether measured under nonallosteric conditions at pH 8.0 or under allosteric conditions at pH 7.0, in the presence of 0.1 or 1 mM ATP. Activities of PFK in Alzheimer's disease and control cells also showed a similar temperature dependence and similar isozyme patterns on column chromatography. These results argue against the existence of significant structural variations of PFK in Alzheimer's disease. PMID- 2977423 TI - Inhibition of T-cell antigen receptor-mediated transmembrane signaling by protein kinase C activation. AB - The murine T-lymphoma cell line LBRM-33 is known to require synergistic signals delivered through the antigen receptor (Ti-CD3) complex, together with interleukin 1 (IL-1), for activation of IL-2 gene expression and IL-2 production. Although 12-O-tetradecanoylphorbol-13-acetate (TPA) was capable of replacing IL-1 as an activating stimulus under certain conditions, biologic studies indicated that TPA failed to synergize with Ti-CD3-dependent stimuli under conditions in which IL-1 was clearly active. Acute exposure to TPA and other active phorbol esters resulted in a concentration-dependent inhibition of the increases in phosphoinositide hydrolysis and intracellular free Ca2+ concentration stimulated by phytohemagglutinin or anti-Ti antibodies. TPA treatment induced no direct alteration of phospholipase C enzymatic activities in LBRM-33 cells. In contrast, both Ti-CD3 cross-linkage and TPA rapidly stimulated the phosphorylation of identical CD3 complex polypeptides, presumably via activation of protein kinase C. Exposure of LBRM-33 cells to TPA resulted in a time-dependent, partial down regulation of surface Ti-CD3 expression. Thus, TPA treatment inhibited the responsiveness of LBRM-33 cells to Ti-CD3-dependent stimuli by inducing an early desensitization of Ti-CD3 receptors, followed by a decrease in membrane receptor expression. These studies indicate that phorbol esters deliver bidirectional signals that both inhibit Ti-CD3-dependent phosphoinositide hydrolysis and augment IL-2 production in LBRM-33 cells. PMID- 2977425 TI - Histaminergic regulation of prolactin secretion: involvement of serotoninergic neurons. AB - The possible involvement of the serotoninergic system in histamine-induced PRL secretion was studied in urethane anesthetized male rats. Intracerebroventricular infusion of histamine (30 micrograms) stimulated PRL secretion 10-fold. This effect was mimicked by the H2-receptor agonist dimaprit (300 micrograms), while the H1-receptor agonist 2-thiazolylethylamine (140 micrograms) had no effect. Pretreatment with the serotonin receptor blockers methysergide (2.5 mg/kg i.p.) or ketanserin (2.5 or 10.0 mg/kg i.p.) reduced the PRL peak response to histamine 75, 54, or 58%, respectively. During serotonin receptor blockade, dimaprit had a stimulatory effect similar to that of histamine, while 2-thiazolylethylamine had no effect. Intraarterial infusion of histamine (420 micrograms) stimulated PRL secretion 6-fold. This effect was mimicked by the H1-receptor agonist 2 thiazolylethylamine (1,900 micrograms), while the H2-receptor agonist dimaprit (3,000 micrograms) had no effect. Pretreatment with methysergide (2.5 mg/kg i.p.) or ketanserin (2.5 or 10.0 mg/kg i.p.) reduced the peak response to histamine 54, 54, or 51% respectively. The effect of histamine was mimicked by 2 thiazolylethylamine, while dimaprit slightly inhibited the PRL secretion. The antiserotoninergic activity of methysergide and ketanserin was demonstrated by their ability to prevent the PRL-releasing effect to serotonin. The effects of methysergide and ketanserin were not due to dopamine-like activity, since none of the drugs affected basal PRL secretion and since the dopamine receptor antagonist pimozide did not prevent the inhibitory effect of methysergide on the histamine induced PRL release. The findings indicate that histamine-stimulated PRL secretion is mediated in part by serotoninergic neurons. PMID- 2977424 TI - The actions of 5-hydroxytryptamine receptor agonists and antagonists at pre- and postjunctional level on the canine saphenous vein. AB - The prejunctional and postjunctional 5-HT receptors of the canine saphenous vein were studied. The release of 3H-noradrenaline (3H-NA) from incubated saphenous vein strips was inhibited by 5-hydroxytryptamine (5-HT) in a concentration dependent way (5-HT concentrations: 0.01, 0.1 and 1.0 mumol.l-1), but not by the selective 5-HT1A agonist 8-hydroxy-dipropylaminotetralin (8-OH-DPAT; 1 and 10 mumol.l-1). The inhibitory effect of 5-HT was antagonized by metitepine and methysergide, but not by yohimbine, (-)-pindolol or ketanserin. In strips preincubated with 5-HT (1.2 mumol.l-1), the fractional release of 3H-NA was slightly reduced (paired experiments). 5-HT and 8-OH-DPAT caused concentration dependent contractions of the saphenous smooth muscle. A parallel shift of the concentration-response curve for 8-OH-DPAT to the right was caused by metitepine and yohimbine, but not by ketanserin. The contractions caused by 5-HT were antagonized by metitepine and yohimbine (parallel displacement of the curves to the right), as well as by ketanserin and methysergide (with a depression of the upper part of the curve). Blockade of alpha-adrenoceptors (due to prazosin plus a low concentration of yohimbine) also resulted in a weak antagonistic effect. Ketanserin and metitepine displaced the noradrenaline concentration-response curve to the right. We conclude that the saphenous vein of the dog is endowed with prejunctional receptors of the 5-HT1 type which can not be classified as belonging either to the 1A or 1B subtype; and that at the postjunctional level 5 HT1 (possibly of the 1D subtype) and 5-HT2 receptors are present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977426 TI - Hypnotic action of flunitrazepam in the rat: does 5-HT mechanism play a role? AB - This study examined whether pharmacological manipulation of serotonergic (5-HT) systems would affect the hypnotic action of flunitrazepam in rats. Flunitrazepam, a potent hypnotic, was used alone or combined with parachlorophenylalanine (pCPA), an inhibitor of the synthesis of 5-HT, 8-OH-DPAT, a 5-HT1A receptor agonist and fluvoxamine, an inhibitor of the reuptake of 5-HT. Flunitrazepam increased the amount of orthodox sleep, the latency of rapid eye movement (REM) sleep and decreased the amount of REM sleep. The drug pCPA decreased the total sleep time and the amount of orthodox and REM sleep. Administration of flunitrazepam to pCPA-pretreated rats induced orthodox sleep in an identical way to that found in the controls. The drug 8-OH-DPAT increased wakefulness and the latency of REM sleep. The association of flunitrazepam with 8-OH-DPAT abolished the increase in waking seen after 8-OH-DPAT alone. In contrast, the combined treatment with flunitrazepam and 8-OH-DPAT resulted in a lengthening of the latency of REM sleep significantly greater than that observed with the same dose of each drug alone. Fluvoxamine increased the latency a decrease the amount of REM sleep. The association of fluvoxamine with flunitrazepam induced a decrease in REM sleep, equal to the sum of the effects of the two drugs alone. Fluvoxamine did not modify the other effects of flunitrazepam. The present experiments demonstrate that the association of pCPA, 8-OH-DPAT and fluvoxamine, did not alter the hypnogenic effect of flunitrazepam. The possibility of an involvement of 5-HT mechanisms in the effect of flunitrazepam on the phasic events in sleep is questionable. PMID- 2977427 TI - The effects of acute and repeated administration of T3 to mice on 5-HT1 and 5-HT2 function in the brain and its influence on the actions of repeated electroconvulsive shock. AB - The effects of the administration of L-triiodothyronine (T3) On the function of 5 HT in the CNS and its influence on the actions of electroconvulsive shock have been examined in mice. A single injection of T3 (100 micrograms/kg) had no effect 24 hr later on either 5-HT1A-mediated hypothermia, induced by 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT; 0.5 mg/kg) or the 5-HT1B-mediated locomotor response to 5-methoxy-3-(1,2,3,6-tetrahydropyridin-4-yl) 1-H-indole (RU 24969; 50 ng i.c.v.). This treatment increased 5-HT2-induced head-twitches, produced by 5 methoxy-N,N'-dimethyltryptamine (5-MeODMT; 2 mg/kg), but did not alter 5-HT2 receptors in the frontal cortex, suggesting that this potentiation was mediated indirectly through a modulatory neurotransmitter. One injection of T3 had no effect on the concentration of 5-HT in the forebrain or mid/hindbrain, but increased 5-HIAA in the latter region. Daily injections of T3 for 10 days attenuated the responses to both 8-OH-DPAT and RU 24969. Furthermore, 5-MeODMT induced head-twitches returned to control values and this was accompanied by a 10% decrease in 5-HT2 receptors in the cortex. Repeated administration of T3 increased levels of 5-HT in mid/hindbrain and concentrations of 5-HIAA both here and in forebrain. Hence, treatment with T3 attenuated the function of 5-HT1A and 5-HT1B receptors, but increased 5-HT2-mediated responses, although the time courses for these effects were different. Triiodothyronine also enhanced the synthesis and turnover of 5-HT in the brain of the mouse. Repeated electroconvulsive shock (90 V, 1 sec) decreased the hypothermia induced by 8-OH DPAT. However, 5-MeODMT-induced head-twitches were enhanced by acute and repeated electroconvulsive shock. Administration of T3 together with electroconvulsive shock did not alter the effects of electroconvulsive shock on 5-HT1A-mediated hypothermia, but markedly potentiated its actions on 5-HT2-mediated responses. These findings provide possible pharmacological evidence for the suggested antidepressant effects of T3 and the potentiation of antidepressant therapy by this thyroid hormone. PMID- 2977428 TI - [Cardiorespiratory findings in top competitive rowers]. PMID- 2977429 TI - [Changes in plasma levels of androgens and SHBG in patients with polycystic ovary syndrome (PCOs) treated with oral contraceptives containing desogestrel]. PMID- 2977430 TI - A study of the capillary permeability in patients with venous hypertension by a new system: the vacuum suction chamber (VSC) device. PMID- 2977431 TI - [Combined therapy of thrombotic thrombocytopenic purpura]. PMID- 2977432 TI - [Principles of the organization of child neurologic services and current problems concerning developmental brain disorders]. PMID- 2977433 TI - [Prevalence of HBsAG during pregnancy at the local health unit in Sassari and initial results of the anti-HBV prophylactic vaccine in newborn infants]. PMID- 2977434 TI - A two-site immunoradiometric assay of proatrial natriuretic factor. Application to tissue extracts. AB - A "two-site" immunoradiometric assay (IRMA) was developed to specifically measure ANF (1-126), the precursor of ANF. This assay is based on the simultaneous use of antibodies against two different antigenic determinants: murine monoclonal antibody (2H2), which recognizes positions 101 through 103 of ANF, is linked to Immunobeads and employed to extract any ANF C-terminal; a second antibody, which is directed against positions 11 through 37, is radioiodinated and allows binding to any C-terminal-2H2-Immunobead material which bears the N-terminal antigenic site. A curvilinear relationship was obtained between radioactivity and the amount of proANF (1.5 to 400 fmol) added. Optimisation of IRMA was determined by the amount of 2H2-Immunobeads and labeled antibody used, incubation time as well as possible interference by both ANF (99-126) and ANF (1-98). Tissue extracts were used to validate the assay. proANF was detected in decreasing amounts in heart atria, heart ventricles, lungs, kidneys and adrenal glands. Its presence was further confirmed by reverse-phase HPLC followed by radioimmunoassay. IRMA is a simple and rapid method for the direct measurement of proANF in tissue extracts and chromatographic fractions. The presence of proANF in tissues strongly suggests local synthesis. PMID- 2977435 TI - Atrial natriuretic factor in the spinal cord of normotensive and hypertensive rats. AB - Atrial natriuretic factor (ANF) was investigated in the rat spinal cord and hypothalamus using two radioimmunoassays. ANF was also quantified in both tissues of Spontaneously Hypertensive Rats and Dahl rats. Spinal cord and hypothalamus were found to be immunoreactive to proANF and its near-NH2- or near-COOH-terminal fragments. A major part of the extracted ANF was a COOH-terminal peptide smaller than or the same as ANF (Ser99-Tyr 126). SHR had higher hypothalamic and spinal cord ANF concentrations than Wistar Kyoto rats, while the Dahl salt-sensitive animals exhibited an increase in spinal cord ANF when compared with the Dahl salt resistant group. The data suggest that spinal cord may produce ANF locally with processing similar to that in hypothalamus. Changes in ANF concentrations occurring during the course of hypertension remain to be further investigated. PMID- 2977436 TI - Presence of atrial natriuretic peptide-like material in guinea pig intestine. AB - Acidic extracts of guinea pig jejunum and colon contain atrial natriuretic peptide-like material (IR-ANP) detected by radioimmunoassay after purification by Sephadex G-50 gel filtration. Immunohistochemical analysis of guinea pig colon also revealed IR-ANP to be located directly beneath the lamina muscularis mucosae. High performance gel permeation chromatography (HP-GPC) and reverse phase high performance chromatography (RP-HPLC) of the IR-ANP showed correspondence to the 15 kD ANP precursor molecule (pro-ANP). No low molecular weight forms of ANP were detected. The extracted pro-ANP could be converted to alpha-ANP-like material by incubation with serum or supernatant of colonic homogenate. These data indicate the intestine to be a further site of ANP synthesis. PMID- 2977438 TI - Determination of spectinomycin dihydrochloride by liquid chromatography with electrochemical detection. AB - Spectinomycin dihydrochloride is determined by liquid chromatography with electrochemical detection. The drug is chromatographed on a reverse-phase Nucleosil C18 column using an eluent containing 0.02 M sodium citrate and 0.0015 M octyl sodium sulfate (pH 6.10 with perchloric acid) and acetonitrile (100:4). Detection is performed using a coulometric detector (porous carbon working electrode) at +0.85 V. The drug and primary degradation product are detectable. Detector response is linear to at least 20 micrograms/ml, which is four times the assay level. The procedure has relative standard deviations of +/- 1.21 to +/- 2.72% for three lots of bulk drug. Sensitivity is greater than 0.1 microgram/ml of spectinomycin (5 ng on column). Repeatability at this level is +/- 4.94%. PMID- 2977437 TI - [Pathogenesis of Bartter's disease]. PMID- 2977439 TI - [Formation of muscle proteins during freezing]. AB - The effect of different conditions on the formation and properties of cryogels prepared by the freezing-thawing procedure from suspensions and solutions of the carp (Cyprinus carpio) myofibrillar proteins was studied. The freezing of water solutions and suspensions of the native myofibrillar proteins resulted in the formation of the structures mainly stabilized by non-covalent bonds. When muscle proteins were denatured prior to the freezing they formed the structures stabilized by both non-covalent and covalent disulfide bonds. PMID- 2977440 TI - [Current problems of phthisio-ophthalmology]. PMID- 2977441 TI - [Importance of preventive fluorography in the early diagnosis of pulmonary tuberculosis in rural population]. PMID- 2977442 TI - First results of an open phase II study with the antidepressant paroxetine. PMID- 2977443 TI - Effect of angiotensin converting enzyme inhibition on renal response to atrial natriuretic factor in rats. AB - Previous studies have shown that atrial natriuretic factor (ANF) inhibits renin secretion whereas cilazapril blocks angiotensin II generation via converting enzyme inhibition. Both agents enhance renal excretory function. The present study was conducted to test whether the renin-angiotension system is involved in the ANF-induced renal effects. ANF was administered to anesthetized normal rats (n = 16) with or without a simultaneous infusion of cilazapril. Single bolus injections of ANF at doses of 2.5 micrograms/kg and 5.0 micrograms/kg significantly decreased mean arterial blood pressure by 6.8 +/- 2.3% and 9.4 +/- 2.2%, respectively. The corresponding increases in glomerular filtration rate were 5.6 +/- 3.7% and 8.4 +/- 2.8%, in absolute sodium excretion were 55.0 +/- 18.5% and 105.2 +/- 39.9%, and in urine flow were 24.8 +/- 9.3% and 35.6 +/- 14.6%. Intravenous infusion of cilazapril (33 micrograms/kg.min) reduced the arterial blood pressure, elevated the glomerular filtration rate and increased sodium and water excretion. The corresponding doses of ANF administration during continuous infusion of cilazapril further decreased blood pressure by 8.3 +/- 1.9% and 10.9 +/- 5.4%, respectively. However, there were no significant changes in the glomerular filtration rate and sodium and water excretion. The failure of ANF to exhibit a renal effect was irrelevant to the lowering blood pressure induced by cilazapril. These results suggest that reduced endogenous angiotensin II generation contributes to the renal, but not the hypotensive, effect of ANF. PMID- 2977444 TI - [Reactive syndromes in Huntington's disease]. PMID- 2977445 TI - [Effect of lymphoid cells of Peyer's patches on the immunologic response in mice]. PMID- 2977446 TI - Carbachol effect on larynx resistance in the cat. PMID- 2977447 TI - [Biopsy with Tru-Cut needle of abdominal subcutaneous fat for diagnosis of amyloidosis. Comparative study with rectal biopsy]. PMID- 2977448 TI - HIV infection: the clinical picture. PMID- 2977449 TI - Dilemmas in the neonatal management of extremely low birth weight babies. PMID- 2977450 TI - Rheumatic disease aspects of leg length inequality. AB - Inequality of leg lengths is associated with low back pain, greater trochanteric bursitis, and degenerative hip disease. The pelvic obliquity that results from a discrepancy of leg lengths may contribute to degenerative changes in the lumbar spine. The diagnosis of leg length inequality may be overlooked. If this occurs, extensive and costly investigations are then often ordered to evaluate these secondary manifestations of leg length inequality. This is unfortunate because a simple shoe lift is all that was needed to provide symptomatic relief for selected patients. PMID- 2977451 TI - An in vitro study of the effects of androgens on the cytoskeleton of ovarian granulosa cells with special reference to actin. AB - Ovarian granulosa cells from small antral follicles from immature rats were cultured in a serum-free medium for 1-6 days with or without the presence of 10( 5) M dehydroepiandrosterone (DHEA) or 10(-5) M-androstenedione (delta 4-A). Control cultures reveal that the cells are flattened and contain many filamentous bundles organized as stress fibers, numerous scattered cytoplasmic actin filaments, microtubules and vimentin. Alpha actinin and myosin were shown by immunocytochemistry to have a punctate pattern along the stress fibers. For the most part, cells exposed to androgens did not flatten; however, they assumed a varied shape and contained fewer stress fibers and actin filaments. Many of these cells did not develop stress fibers and those that did develop were fewer in number and displayed--actinin and myosin in a punctate pattern. Microtubules and vimentin filaments remained unaltered when compared to controls. It is believed that the deficiency of actin filaments, coupled with certain other degenerative changes which express themselves in other cellular compartments, leads to an early atresia of the granulosa cell cultured in high concentrations of androgens. PMID- 2977452 TI - Immunological status of nude mice engrafted with allogeneic or syngeneic thymuses. AB - Restoration of T-cell functions and changes in autoantibody production were studied in BALB/c nu/nu (nude) mice engrafted with syngeneic (BALB/c) or allogeneic (C57BL/6J) thymuses across major histocompatability barriers. T-cell functions, including mitogen responses and antibody production to sheep red blood cells (SRBC), were restored in nude mice engrafted with either allogeneic or syngeneic thymuses. Alloreactivity was evaluated by analysis of the pattern of skin allograft rejection, generation of alloreactive cytotoxic T-lymphocytes (CTLs), or quantitation of mixed-lymphocyte reaction (MLR). BABL/c nude mice engrafted with thymuses from newborn C57BL/6J mice accepted the skin from either thymus donor-type mice or from host-type mice. By contrast, such thymic chimeras rejected skin grafts from a third-party donor. CTLs from nude mice engrafted with C57BL/6J thymuses were cytotoxic to target cells of the third party but not to target cells of the host-type or of the thymus-type. In the MLR assay, spleen cells of nude mice engrafted with C57BL/6J thymuses responded vigorously to third party cells and only slightly to cells of the thymus-type. Low levels of serum IgG and high titers of IgM antibodies to nuclear antigens (but not dsDNA) or skin basal cells were also found in nude mice. Antibodies to both nuclear antigens and skin basal cells disappeared after transplantation of syngeneic thymuses, but not after transplantation of allogeneic thymuses. By contrast, serum IgG levels were restored to normal in nude mice engrafted with either syngeneic or allogeneic thymuses. These results suggest that either HLA-matched or HLA-mismatched thymus grafts may become a viable treatment for certain patients with T cell deficiencies associated with deficient development or maintenance of thymic structure and/or function. PMID- 2977453 TI - Growth of single T cells and single thymocytes in a high cloning efficiency filler-cell free microculture system. AB - A high cloning-efficiency microculture system is described in which single T cells, stimulated to divide by phorbol ester and calcium ionophore, grow rapidly under the influence of purified growth factors in the absence of other cells. The kinetics of clonal growth has been monitored over a five day period by phase contrast microscopy. Mature peripheral T cells, and mature subpopulations from the thymus, responded with a cloning efficiency over 80%; they required IL-2 as a minimum but several other factors enhanced growth. Ly2+L3T4- thymocytes (mean doubling time 10.4 hr) grew more rapidly than Ly2-L3T4+ thymocytes (mean doubling time 15.2 hr). Early (Ly2-L3T4-) thymocytes responded with a cloning efficiency of 60%; their efficient growth was dependent on both IL-1 and IL-2. The typical Ly2+L3T4+ cortical thymocyte did not grow under these conditions. PMID- 2977454 TI - Antigen processing: an interim report. PMID- 2977455 TI - Novel mechanisms controlling arginine metabolism in Neurospora. PMID- 2977456 TI - The E. coli Rho protein: an ATPase that terminates transcription. PMID- 2977457 TI - This and that: on transport and translation. PMID- 2977459 TI - Intramuscular versus subcutaneous injection for hepatitis B vaccine. PMID- 2977458 TI - The metabolism of propranolol (ICI 45,520, Inderal) and xamoterol (ICI 118,587, Corwin) by isolated rat hepatocytes: in vivo-in vitro correlations. AB - 1. The metabolism of two compounds which undergo predominantly Phase I (propranolol) and Phase II (xamoterol) metabolism in vivo has been studied in isolated rat hepatocytes. 2. Propranolol was rapidly metabolized by rat hepatocytes to a number of metabolites which correlated well with those observed in vivo. The effect of saturable metabolism on the in vitro clearance of propranolol at high substrate concentrations was very similar to the changes observed in vivo. 3. Xamoterol was metabolized by rat hepatocytes to produce mainly xamoterol glucuronide, with the sulphate conjugate of xamoterol representing a minor component. The low rate of formation of xamoterol sulphate is probably due to the low affinity of xamoterol for the sulphotransferase enzyme, since supplementation with inorganic sulphate did not significantly alter the rate of sulphation; the sulphotransferase system of these hepatocytes was however shown to be active in the metabolism of phenol. 4. The correlations observed between the known routes of metabolism of propranolol and xamoterol in vivo and those observed in isolated hepatocytes support the utility of isolated hepatocytes as a predictive model of metabolic events in vivo. PMID- 2977460 TI - [Epidemiologic aspects of early latent syphilis]. PMID- 2977461 TI - Three years' experience of the east New Britain project for the disabled. PMID- 2977462 TI - [Backache as an occupational disease?]. PMID- 2977463 TI - [The noncorrespondence of the geographic ranges of the host and the causative agent]. AB - For the first time the experimental and field studies of Leptospira infections, carried out over the period of 6 years, have revealed that the habitat of striped field mice (Apodemus agrarius Pallas 1778) serving as a host for Leptospira pomona, serovar mozdok, is much wider than the habitat of the infective agent proper. The presence of an animal species highly sensitive to a definite leptospiral serovar and serving as its reservoir at a given locality cannot be regarded as a proof of the presence of the epizootic process without bacteriological confirmation. But, in the absence of homologous Leptospira carriage, intensive leptospiral seroconversion can be attributed to a population other than that of the host. PMID- 2977464 TI - [The possibility of using the demonstration of Shigella antigens in the study of the epidemic process in dysentery]. AB - The detection of Shigella antigens in feces in accordance with the results of the passive hemagglutination test is more effective than the bacteriological method for finding out persons infected with Shigella, especially in the absence of pronounced clinical manifestations of dysentery, and can be used in the study of the mechanism of the epidemic process. The examination of two groups, each consisting of permanent and temporary members, has revealed that the use of this method for detection of the antigen makes it possible to perform the epidemiological analysis of morbidity and, in particular, to determine the source(s) of infection. PMID- 2977465 TI - [The results of a seroepidemiologic study of Newcastle dysentery]. AB - Differences in the monthly distribution in the number of seropositive individuals among children and adults in years with different intensity of the epidemic process have been revealed. Immunity in cases of dysentery caused by S. newcastle reflects the yearly and seasonal activation of the epidemic process in this Shigella infection both in children and adults, the seroconversion characteristics observed in the year of a high morbidity level being a more objective criterion indicating the beginning of the activation of the epidemic process in dysentery caused by S. newcastle in comparison with the commonly registered morbidity level. PMID- 2977467 TI - [Diagnostic characteristics of acute neurologic pathology at the prehospital stage]. AB - Clinically verified files of 1436 patients with 12 most frequent forms of acute neurological pathology were studied for the rate and causes of prehospital diagnostic errors. The information obtained before the patient's admission was evidenced to be insufficient for the reliable differentiation of the ailment nature. In cases when precise diagnosis appears impossible at the site of first medical aid, its efficiency can be increased by correct choice of special hospital. PMID- 2977466 TI - [The frequency of detection of markers of hepatitis virus B infection among the inhabitants of Rustavi, the Georgian SSR]. AB - Blood serum samples from 1,087 patients with acute viral hepatitis were studied. HBsAg was detected in 36.6% of cases. The study of anti-HBc IgM made it possible to diagnose hepatitis B in 6.6% of cases. The study of blood serum samples from 362 donors, 2,356 pregnant women and 163 medical workers in Rustavi for the presence of the markers of hepatitis B infection revealed a wide spread of hepatitis B in Rustavi. PMID- 2977468 TI - [Hormone content of the blood in neurotic patients]. AB - Radioimmunoassay was applied to study the hormones (corticotropin, cortisol, T3, T4, testosterone, prolactin, follicle-stimulating and luteinizing hormones) in 52 male neurotic patients and 120 healthy donors in Arkhangelsk city, as compared to latitudinal mean values. In patients, prolactin, cortisol, thyroid hormones and gonadotropins were increased by 82.7%, 48.7%, 41%, 52.6%, respectively. Patients also had lower production of ACTH, higher blood contents of T3, prolactin and testosterone. Unidirectional shifts were detected in hormonal systems of neurotic patients, with respect to the time of the disease onset and the duration of last exacerbation: ACTH secretion increased with reduced response of adrenal cortisol production, compensatory increase in thyroid functions, redistribution of gonadotropic fractions increasing the FSH/LH ratio, decrease in testosterone production. At the initial stages prolactin secretion increased to reach later its plateau. PMID- 2977469 TI - Gastroschisis: prenatal diagnosis and management. AB - Ten fetuses with gastroschisis on whom a prenatal sonographic diagnosis and evaluation were available and who delivered at the University of California, San Francisco, were evaluated retrospectively. Six fetuses were delivered vaginally without mortality and minimal or absent morbidity. Four were delivered abdominally, 2 for unrelated obstetrical indications. A 3rd fetus had a suspected prenatal bowel perforation and was the sole mortality. Only 1 mother underwent primary cesarean section, specifically due to the fetal gastroschisis. Umbilical cord pH values were normal in those delivered vaginally. With careful ultrasound examination and immediate neonatal surgical capability in a level III perinatal center, vaginal delivery of fetuses with gastroschisis can be associated with excellent outcome. A multinational randomized study of the delivery mode of fetuses with gastroschisis is necessary and appropriate at this time. PMID- 2977470 TI - The effect of hormonal imprinting on 3H-steroid incorporation of Tetrahymena pyriformis. AB - Steroid hormones are incorporated by Tetrahymena, and appear intracellularly in both cytoplasmic and intranuclear localizations. At primary interaction with the steroid, incorporation was the greatest at 10 min, whereas at the second interaction it tended to increase with time. The Tetrahymena cells preexposed to a steroid incorporated a greater amount of it at the second exposure, owing presumably to the induction of steroid receptors at the first interaction. PMID- 2977471 TI - Some epidemiological characteristics of Down's syndrome in Hungary. AB - Down's syndrome seems to be an adequate indicator condition of germinal numerical chromosome mutation. The true birth prevalence of Down's syndrome was 1.2 +/- 0.1 per 1000 total births in Hungary 1973-1982. The rate of undernotification and misdiagnosis was 25% and 5%, resp. A significant deviation was detected in the occurrence of Down's syndrome in twenty territorial-administrative units of Hungary due to the different ascertainment. The maximum figures may represent the true birth prevalence. No significant monthly variation, i.e., seasonality was found in the birth of Down's syndrome. A Surveillance of Down's syndrome was established in 1980 based on the Hungarian Congenital Malformation Registry. PMID- 2977472 TI - The impact experiment as a source of information about the mechanism of origin of lower leg trauma. PMID- 2977473 TI - Genetic basis of virulence in dermatophytes. PMID- 2977474 TI - Mutants of Trichophyton equinum. PMID- 2977475 TI - Somatometry of the Czechoslovak Spartakiade 1985. PMID- 2977476 TI - [Development of the enzyme systems of the human embryonic and fetal heart]. PMID- 2977477 TI - Blood supply of the rat liver in scanning electron microscopy. PMID- 2977478 TI - Diagnostic significance of serum proteinase activity ("cathepsin B-like" activity) testing by amino acid derivatives of 7-amino-4-methylcoumarine. PMID- 2977479 TI - Changes of serum proteinase "cathepsin B-like" activity in patients with lung cancer and plasmocytoma after therapy. PMID- 2977480 TI - Leukotrienes and other lipoxygenase metabolites of arachidonic acid: their role in edema, ischemia and demyelination of the central nervous system. PMID- 2977481 TI - Blood serum vitamin E and A levels during peroral therapy of periodontal diseases with zinc sulphate. PMID- 2977482 TI - Peritumorous cellular reaction in Grawitz carcinoma. PMID- 2977483 TI - Disorders of metabolism of magnesium and its clinical and laboratory features. PMID- 2977484 TI - Determination of HLA phenotypes of relatives--a contribution to the assessment of paternity probability. I. PMID- 2977485 TI - Determination of HLA phenotypes of relatives--a contribution to the assessment of paternity probability. II. PMID- 2977486 TI - Age dependence of the body mass indices. PMID- 2977487 TI - Exclusion and inclusion of paternity: 1976-1985. PMID- 2977488 TI - HLA antibodies in pregnant women. PMID- 2977489 TI - More attention to meningococci. PMID- 2977490 TI - Microbial eczema--bacterial findings, skin tests, state of natural resistance and of humoral immunity. PMID- 2977492 TI - Diagnostics and current possibilities of treating Sarcoma idiopathicum multiplex haemorrhagicum Kaposi. PMID- 2977491 TI - Therapy of microbial eczema with staphylococcal vaccine. PMID- 2977493 TI - Quantitative determination of surfactants in aqueous eluents from the horny layer of the epidermis. PMID- 2977494 TI - Application of biostimulative effects of He-Ne laser in the therapy of crural ulcers. Ultrastructural findings in irradiated tissue. PMID- 2977495 TI - Suppression of lactation with Terguride. PMID- 2977496 TI - Measurement of regional uteroplacental blood flow. PMID- 2977497 TI - Pregnancy-induced hypertension and its influence on uteroplacental blood flow. PMID- 2977498 TI - Changes in uteroplacental blood flow in relation to intrauterine hazard to fetus. PMID- 2977499 TI - Extrasphincteric posterior perineal approach to the organs of the lesser pelvis. PMID- 2977500 TI - [A new surgical technic for congenital anorectal anomalies]. PMID- 2977501 TI - [Histomorphological mucosal changes in laryngeal carcinoma]. PMID- 2977502 TI - [Histomorphological changes in the mucosa in chronic laryngitis]. PMID- 2977503 TI - Radiological findings in postoperative discitis. PMID- 2977504 TI - Body mass indices--prediction efficiency. PMID- 2977506 TI - Lipoprotein X in the cholestatic syndrome. PMID- 2977505 TI - [Echography in the diagnosis of urothelial tumors]. PMID- 2977507 TI - Diabetic nephropathy. PMID- 2977508 TI - Actual possibilities of examination of sinoatrial node function. PMID- 2977509 TI - Rupture of the heart and acute myocardial infarction. PMID- 2977510 TI - Perforating keratoplasty in patients with keratoconus. PMID- 2977511 TI - Our experience with faradization of urinary bladder in enuretic children. PMID- 2977512 TI - Collective education at the university and its socially psychological foundations applied to the conditions of the faculties of medicine. PMID- 2977513 TI - Teeth extraction. Reasons--procedures--circumstances. PMID- 2977514 TI - Linear regression of body mass on stature and chest circumference. PMID- 2977515 TI - [Total activity and isofermentation of lactate dehydrogenases in peripheral lymph in relation to lymphatic flow]. PMID- 2977516 TI - [Activity of mitochondrial enzymes in patients with acute viral hepatitis B]. PMID- 2977517 TI - [Circulatory homeostasis in patients with hypertension during acute and chronic blockade of alpha 1-adrenoreceptors]. PMID- 2977518 TI - [Cardiovascular effects of aspartic acid]. PMID- 2977519 TI - [Hormonal parameters in the polycystic ovary syndrome]. PMID- 2977520 TI - [Various treatment regimens in infertility using bromoergocryptine in women with hyperprolactinemia]. PMID- 2977521 TI - Body dimensions of miners in Ostrava. PMID- 2977522 TI - Bioincompatibility of dialysis membranes: factor H binding correlates inversely with complement activation indicating a local imbalance of involved proteases/anti-proteases. PMID- 2977523 TI - Neutrophil elastase, thrombin and plasmin in septic shock. PMID- 2977524 TI - Atrial natriuretic peptide (ANP)-induced increase in capillary albumin and water flux. PMID- 2977525 TI - Correlation of laser-Doppler-velocity measurements and endothelial cell shape in a stenosed dog aorta. AB - Laser-Doppler-velocity measurements were carried out in an elastic 1:1 true-to scale silicone rubber model of a dog aorta with stenosis. The model was constructed from a cast of a severely stenosed dog aorta (71% of its area). The stenosis in the dog aorta was prepared by wrapping a cotton band around the aorta. This band was tightened until the presence of a thrill or a bruit was felt distal to the band. Twelve weeks later the animal was sacrificed and a cast was prepared from the aorta. From this vascular cast, the cross-sectional area was calculated. Endothelial cell geometry and orientation was studied using computerized analysis to determine the cell area and shape index. An elastic silicone rubber model was prepared from the cast to measure the velocity profiles and to estimate the local wall shear stress. Velocity measurements were done at steady and pulsatile flow using a Newtonian aqueous-glycerol solution and a non Newtonian blood-like fluid. From those velocity measurements the velocity gradients near the wall were determined and the shear stress calculated. The flow distal to the stenosis separates from the wall at physiological conditions. The endothelial cells are smaller and more elongated in the throat; distal to the stenosis they are larger and rounder. The shape index distribution along the stenosed aorta is correlated with the level of wall shear stress. It is shown that even low changes in the wall shear stress have an influence on the orientation of the endothelial cells. PMID- 2977526 TI - [Clinical effect of hotyuekkito (buzhongyiqitang) on symptoms due to renal ptosis and stress incontinence]. AB - Eleven patients with lumbago due to renal ptosis, and 23 patients with stress incontinence were treated with Tsumura Hotyuekkito (Buzhongyiqitang), an old Chinese prescription, 7.5 g, three times a day. The subjective symptoms were improved in 9 cases (82%) of lumbago and 18 cases (78%) of stress incontinence. No side effects were observed. PMID- 2977527 TI - [Endocrine therapy for prostatic carcinoma--the clinical trial to compare the efficacy of LH-RH analogue, ICI 118630 (Zoladex) with castration or estrogen]. AB - A multicenter randomized clinical trial was carried out between May, 1986 and May, 1987 involving 82 patients with stage B-D prostatic carcinoma from 29 centers. The clinical efficacy, endocrine effect, safety and usefulness of the luteinizing hormone-releasing hormone (LH-RH) analogue and other endocrine manipulations in the treatment of prostatic carcinoma. Zoladex depot containing 3.6 mg of ICI 118,630, an LH-RH analogue, was administered every four weeks 3 times in total. Patients in the control group received either 300 mg of diethylstilbestrol diphosphate orally daily for 12 weeks or orchidectomy. An antitumor effect (CR + PR) was observed in 21 of the 33 patients (63.6%) in the Zoladex group and in 22 of the 33 (66.7%) in the control group, showing no significant difference between the two groups. There was no significant difference in overall subjective response either; 21 of the 24 (87.5%) in the Zoladex group and 24 of the 30 (80.0%) in the control group. In both groups, 100% endocrine effect was obtained as shown by achievement of castration in all patients. Adverse reactions were observed in 14 of the 39 (35.9%) patients treated with Zoladex as compared with 19 of the 34 (55.9%) control patients, resulting in no significant difference in the incidence between the two groups. These adverse reactions were not so severe as to require withdrawal from the study. In both groups, the treatment was assessed as slightly or more useful in 29 of the 33 (87.9%) patients. From these results, it is concluded that Zoladex, 3.6 mg depot, is a useful drug for treatment of prostatic cancer, having clinical efficacy and endocrine effects comparable to those of the conventional endocrine manipulations, being safe, and causing less physiological and psychological pain. PMID- 2977528 TI - Filter-supported preparation of lambda phage DNA. AB - A rapid and simple method is described for the isolation of DNA from phage lambda which requires neither special equipment nor expensive material such as cesium chloride for ultracentrifugation nor extractions with organic solvents or ethanol precipitation. Microgram quantities of lambda DNA are obtained in less than 2 h from 90-mm plate lysates or 5-ml liquid cultures. The method allows the simultaneous isolation of large numbers of probes, e.g., clones from phage libraries. Lambda phages are precipitated by polyethylene glycol/sodium chloride and recovered by low speed centrifugation onto glass fiber filters positioned in disposable syringes. The DNA of phages is released by a 50% formamide/4 M sodium perchlorate solution, washed in filter-bound form, eluted with a small volume of low-salt buffer or water, and finally recovered by centrifugation. Comparison of the DNA isolated by this method with that obtained by two conventional procedures reveals both a similar recovery and a similar suitability for restriction enzyme digestion and subcloning. PMID- 2977529 TI - Preparation of bacteriophage lambda DNA using the TL-100 ultracentrifuge. AB - A procedure for the preparation of DNA from bacteriophage lambda is described, using the Beckman TL-100 bench-top ultracentrifuge. The procedure involves growth of phage in agar plates, precipitation with polyethylene glycol, and a single centrifugation in cesium chloride under conditions that disrupt the phage coat. The method avoids the use of enzymes, ion exchange resins, and phenol. It can be completed in less than a day. The resulting DNA is of good purity and is easily cuttable by restriction enzymes. PMID- 2977530 TI - [Concentration of beta-endorphin and met-enkephalin in the blood of surgical patients in critical states]. PMID- 2977531 TI - [The operative management of the anesthesiology-resuscitation service of a hospital based on a systems approach and computerization]. PMID- 2977532 TI - [Fecal surveys in pastured sheep and the occurrence of Cysticercus tenuicollis in slaughtered sheep]. AB - Out of 4 fenced pastures, 2 stationary flocks and 3 wandering flocks with approx. 2,000 mother sheep, 962 samples of faeces were checked for the occurrence of gastro-intestinal nematodes and lungworms. Irrespective of the way the sheep are kept, an infestation incidence of 11.1 to 100% with gastro-intestinal nematodes was found. In one flock even after two treatments 66.7% were still infected; therefore, suspect methods of treatment have to be avoided and a constant change of anthelmintics is absolutely essential. Dictyocaulus filaria occurred in 6 out of 9 flocks and was easy to be treated chemotherapeutically, whereas the protostrongyles could be reduced but were not eliminated totally. - 785 out of 4,710 slaughter sheep (= 16.7%) harboured metacestodes of Taenia hydatigena in the omentum majus. In most cases one or two specimens (= 66.6 i.e. 17.6%) of Cysticercus tenuicollis could be found, once there were even 117. 1.332 cysticercus (= 78.1%) showed ready to contract and according to these and other morphological criteria, a total of 1,416 (= 83%) were regarded to be vital and able to evoke an infection. Relating to practice, the sheep dog should always be treated at the same time as deworming of the flock takes place. PMID- 2977533 TI - [Therapeutic effectiveness of the gonadotropin releasing hormone analogue D-Trp-6 LHRH in the treatment of precocious puberty]. AB - Efficacy and safety of a superactive gonadotrophin-releasing hormone analogue (D Trp-6-LHRH, decapeptyl) was assessed in seven girls, aged two to seven years with precocious puberty (four idiopathic, two associated to McCune-Albright syndrome and one with myelomeningocele). A further case of a seven year old male was treated with a delayed release formulation of the analogue combined with cyproterone acetate for the first two weeks of therapy. Three cases had previously been unsuccessfully treated with cyproterone acetate (2% or medroxyprogesterone (1) but this conventional therapy was discontinued three months prior entering the trial. Decapeptyl was given at a dose of 100 mcg sc daily for 10 days, followed by a dose of 1-2 mcg/kg daily thereafter. One patient required a sustained dose of 7 mcg/kg for effective control. The male patient received a biodegradable depot preparation of decapeptyl at a dose of 1.5 mg IM monthly, calculated to release circa 100 mcg of the analogue daily. He was also treated with cyproterone acetate. 100 mg/m2/day 15 days before and for two weeks during decapeptyl administration. Treatment was well tolerated without significant side effects. After six months on treatment, both LH and FSH levels were undetectable and showed no response to LHRH. Plasma oestradiol levels were reduced from 230 +/- 40 to 13 +/- 4 pg/ml. Total testosterone in the male patient was suppressed from 7 ng/ml to undetectable levels. This was accompanied by a cessation in the progression of the development of secondary sexual characteristics. All female patients developed amenorrhoea and their Tanner stage regressed to I or prepubertad after treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977534 TI - Evoked effects of PGE2 and PGA2 on lipid fluidity and Ca2+-stimulated ATPase of Walker-256 tumor microsomal membranes. PMID- 2977535 TI - Lymphocyte activation: modulatory effects mediated by interactions between cell adhesion molecules. PMID- 2977536 TI - Effect of ADP-ribosyl transferase inhibitors on the survival of human lymphocytes after exposure to different DNA-damaging agents. PMID- 2977537 TI - Cilofungin (LY121019) inhibits Candida albicans (1-3)-beta-D-glucan synthase activity. AB - Cilofungin (LY121019) inhibited Candida albicans growth and activity of (1-3) beta-glucan synthase, for which it was a noncompetitive inhibitor with a Ki-app of 2.5 microM. Cilofungin had no effect on chitin synthase activity. Based on these and other data, it seems likely that cilofungin inhibits fungal growth by inhibiting (1-3)-beta-glucan synthase activity. PMID- 2977538 TI - Effects of atrial natriuretic factor (ANF) on rat testicular steroidogenesis in vitro. AB - The aim of this study was to evaluate the effects of rat atrial-peptide type II (rAP-II) on testicular steroidogenesis by isolated adult rat Leydig cells. rAP-II stimulates testosterone secretion. The maximal stimulatory effects of rAP-II on testosterone production occurred at the dose of 10(-11) M. At the same dose this peptide stimulates androstenedione and dehydroepiandrosterone-sulphate production, two important testosterone precursors of delta-4 and delta-5 steroidogenetic pathways, respectively. At higher doses, 10(-9) and 10(-7) M, the stimulatory effect of rAP-II on testosterone secretion is strongly declined, on androstenedione secretion is abolished, whereas on dehydroepiandrosterone sulphate secretion remains unaffected. These data suggest that rAP-II at low doses exerts a stimulatory effect on the early steroidogenetic step by isolated adult rat Leydig cells. At higher doses this peptide seems to influence the testicular steroidogenesis, probably exerting a limiting reaction step involving the 3-beta-OH-steroid-dehydrogenase activity. PMID- 2977539 TI - [Percutaneous pulmonary valvuloplasty. Short-term and long-term results]. AB - Percutaneous angioplasty was performed in twenty consecutive patients, with congenital pulmonary valve stenosis. Ages ranged from eight months to thirty-two years (mean 9.5 years old). We achieved a valvular gradient dropping from 91 +/- 39 to 19 +/- 11 mm Hg (P less than 0.001) in early post angioplasty level and it was practically unchanged at three months and one year later. (19 +/- 12, 19 +/- 17 mm Hg) (P less than 0.001). Similar change was observed in the right ventricle systolic pressure which was diminished in a progressive way during the follow-up from 113 +/- 37 to 39 +/- 35 (P less than 0.001), 59 +/- 18 and 53 +/- 25 mm Hg (P less than 0.001) immediately, three months and one year later, respectively. The ratio right ventricle systolic pressure/left ventricle was diminished from 0.96 +/- 29 to 0.63 +/- 0.35 in the early post angioplasty period and later from 0.50 +/- 0.16 and 0.44 +/- 0.22 (P less than 0.001). Only one case had restenosis one year later and we repeated the angioplasty with good results. Most of the patients are asymptomatic, the pulmonary murmur features changed. We observed improvement in electrocardiographic and echocardiographic signs. One patient died of anesthetic complications. The remainder of patients did not have severe complications and they were discharged from 48 to 72 hours after angioplasty. In conclusion, valvuloplasty is an effective procedure in a short and long term basis. We considered valvuloplasty in congenital pulmonary valve stenosis the treatment of choice in this group of patients. PMID- 2977541 TI - Honesty--the best policy. PMID- 2977542 TI - Dental care for AIDS virus carriers. PMID- 2977540 TI - Antihypertensive activity of a new calcium channel blocker LF 2.0254, in spontaneously hypertensive rats: a comparison with nifedipine and nicardipine. AB - LF 2.0254 is a new dihydropyridine calcium channel blocker with a long-lasting antihypertensive action in spontaneously hypertensive rats. In acute experiments, LF 2.0254 was shown to be more potent than nifedipine and nicardipine in reducing blood pressure after i.v. and oral administration with less tachycardia. The hypotensive effects of LF 2.0254 (10 mg/kg) lasted for greater than 24 hr, whereas the blood pressure-lowering effects of nifedipine and nicardipine (10 mg/kg) had disappeared after 6 hr. LF 2.0254 given orally once a day at 10 mg/kg for 14 days to spontaneously hypertensive rats, led to a marked and persistent decrease in blood pressure, with no significant changes in heart rate. In an attempt to prevent the development of hypertension, LF 2.0254 (3 and 10 mg/kg) was given once a day for 8 weeks to 5 week-old SHR. On the last day of treatment, LF 2.0254 had markedly reduced blood pressure over a 24 hr period, however, there was no reduction of left ventricular hypertrophy, since body weight and left ventricular weight were not significantly different between control and treated groups. PMID- 2977543 TI - Long-term clinical and angiographic results following percutaneous transluminal coronary angioplasty. AB - The long-term results of percutaneous transluminal coronary angioplasty were evaluated in our first 42 patients, who had initial successful angioplasty and no restenosis on angiogram performed four months after angioplasty. Evaluation included repeat follow-up angiogram performed approximately two years after angioplasty in 22 of the 42 patients. All 42 patients had single vessel disease with severe angina pectoris refractory to medical treatment before angioplasty; at a mean follow-up of 28 months after angioplasty, 37 (88%) were asymptomatic. No patient died or underwent coronary artery bypass graft surgery during the follow-up period, although three patients (7%) suffered a myocardial infarction due to occlusion of a nondilated artery. In the 22 cases studied at a mean of 28 months after angioplasty, angiogram showed that successful coronary artery dilatation remained in all cases, with no deterioration of a dilated lesion between the four and 28 month angiograms. Progression of atherosclerosis, however, did occur in five of the 22 patients (23%), with development of either stenosis or occlusion of a non-dilated vessel. This study suggests that the development of restonsis between four and 28 months after percutaneous transluminal coronary angioplasty is unlikely. The results suggest an excellent long-term prognosis after angioplasty, in patients who have no evidence of restenosis at four months after an initially successful dilatation. PMID- 2977544 TI - Enhancing social skills and self-perceptions of physically disabled young adults. Assertiveness training versus discussion groups. PMID- 2977545 TI - Cardiovascular effects of the new calcium antagonist isradipine and of diltiazem in anesthetized open-chest dogs. AB - Cardiovascular effects of the new calcium antagonist, isradipine (PN 200-110), were compared with those of diltiazem in anesthetized open-chest dogs. Isradipine 5 micrograms/kg i.v. produced significant decreases in systolic, diastolic and mean aortic blood pressure (AoP) concomitant with a decrease in mean renal blood flow (RBF) and increases in mean vertebral blood flow (VBF), mean coronary blood flow (CBF) and left ventricular dP/dt (LVdP/dt), but almost unchanged heart rate (HR) and left ventricular enddiastolic pressure (LVEDP). Diltiazem 300 micrograms/kg i.v. also produced decreases in AoP and RBF and increases in AoF, VBF and CBF. LVdP/dt and LVEDP were not significantly changed, but HR was decreased by this drug. Duration of increase in AoF, VBF and CBF was significantly longer in isradipine than in diltiazem. The decrease of coronary vascular resistance relative to total peripheral resistance was significantly greater than 1.0 for diltiazem, but not for isradipine. Results indicate that isradipine produces effects on AoP, AoF, VBF, CBF, RBF and LVEDP similar to diltiazem and the drug increases LVdP/dt without a decrease in HR in contrast to diltiazem, and that the effects of isradipine were long sustained when compared with those of diltiazem. PMID- 2977546 TI - Pressure effects on the interactions of the sarcoplasmic reticulum calcium transport enzyme with calcium and dinitrophenyl phosphate. AB - The effect of hydrostatic pressure on the calcium-dependent hydrolysis of dinitrophenyl phosphate by the sarcoplasmic calcium transport enzyme has been studied. The magnesium dinitrophenyl phosphate complex is the true substrate of the enzyme (K = 7000 M-1) by which it is hydrolyzed at 20 degrees C with a turnover rate of 4 s-1. Activation by calcium ions occurs between 0.1 and 1 microM as observed for ATP hydrolysis. The activation volume of the enzyme saturated with both ligands exhibits pronounced pressure-dependence, rising from 25 ml/mol at atmospheric pressure to 80 ml/mol at 100 MPa. The apparent binding volumes for magnesium dinitrophenyl phosphate and calcium are likewise pressure dependent. The volume changes connected with the binding of magnesium dinitrophenyl phosphate is quite small approaching zero at 100 MPa. The apparent binding volume for calcium greatly increases with pressure from 35 ml/mol at atmospheric pressure to 150 ml/mol at 70 MPa. A nearly constant binding volume of approximately 40 ml/mol results if the effect of pressure on the respective rate constants that contribute to the apparent binding constant, is taken into account. The pressure-dependence of enzyme activity at subsaturating calcium concentrations yields an activation volume of 250 ml/mol related to the rate of calcium binding indicating the occurrence of a transient large volume expansion of the enzyme complex. The volume changes observed for the calcium-dependent interaction of the enzyme with magnesium dinitrophenyl phosphate well agree with that found for magnesium p-nitrophenyl phosphate (W. Hasselbach and L. Stephan,Z. Naturforsch. 42 c, 641-652 (1987)) indicating that the found volume changes are intrinsic properties of the transport enzyme, independent of the respective energy donor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977547 TI - [Canadian experiences with AIDS and HIV infection]. PMID- 2977548 TI - [Perspectives on the AIDS epidemic: experience in the United States]. PMID- 2977549 TI - [AIDS in Mexico: trends and projections]. PMID- 2977551 TI - [AIDS in Brazil, 1982-1988]. PMID- 2977550 TI - [AIDS in Colombia]. PMID- 2977552 TI - [Prevalence of HIV-1 infection in viral hepatitis patients with a history of intravenous drug abuse]. PMID- 2977553 TI - [Epidemiology of AIDS and HIV infection in the Caribbean]. PMID- 2977555 TI - [Prevalence of infection with human immunodeficiency virus in Cuba]. PMID- 2977554 TI - [Perinatal transmission of the human immunodeficiency virus]. PMID- 2977556 TI - [Immunopathogenic aspects of infection by the human immunodeficiency virus in Venezuela]. PMID- 2977557 TI - [Retroviruses in the Caribbean]. PMID- 2977559 TI - [Public knowledge about AIDS in Brazil, the Dominican Republic, Haiti and Mexico]. PMID- 2977558 TI - [Simple procedure for obtaining large quantities of HIV antigens for serodiagnosis]. PMID- 2977560 TI - [AIDS: social, legal and ethical aspects of the "third epidemic"]. PMID- 2977561 TI - [Prevention of the transmission of HIV through blood. The Mexican experience]. PMID- 2977562 TI - [The Canadian national AIDS program]. PMID- 2977563 TI - [The prognosis for AIDS has been exaggerated]. PMID- 2977564 TI - [Prognosis on the number of AIDS cases, United States of America]. PMID- 2977565 TI - [Projections on AIDS: perspective from Jamaica]. PMID- 2977566 TI - [The changing pattern of HIV transmission in the Caribbean]. PMID- 2977567 TI - [What's the real reason for the prognosis on AIDS?]. PMID- 2977568 TI - Lysosphingolipids and mitochondrial function. II. Deleterious effects of sphingosylphosphorylcholine. AB - Psychosine, sphingosylphosphorylcholine (52-104 microM), and other glycosphingolipids stimulate mitochondrial respiration (up to 500%) and inhibit oxidative phosphorylation to varying degrees. Above 104 microM these functions as well as uptake of Ca2+ are prevented. At 104 microM sphingosylphosphorylcholine inhibits the mitochondrial ATPase reaction in submitochondrial particles by 48%. Both sphingosylphosphorylcholine and psychosine enhance the active phosphate dependent swelling of mitochondria. Passive swelling occurs in the presence of rotenone (when swelling does not normally occur) and under hypotonic conditions. A direct interaction of sphingosylphosphorylcholine with membranes is demonstrated by a discharge of the proton gradient across mitochondrial membranes, hemolysis of red blood cells, and binding to inner and outer mitochondrial membranes. Thus lysosphingolipids bind strongly to mitochondrial membranes and markedly alter mitochondrial function. This alteration would affect the ATP levels, thereby altering a wide range of ATP-dependent cellular functions. These results offer a partial explanation for the pathogenesis of representative lysosomal storage diseases. PMID- 2977569 TI - Communication aids provision 1983-1986. PMID- 2977570 TI - Update on percutaneous transluminal coronary angioplasty. AB - Percutaneous transluminal coronary angioplasty was first performed in a human coronary artery in 1977. In this article, we review the possible mechanisms of angioplasty, current indications for angioplasty, and complications of angioplasty including acute closure and restenosis. PMID- 2977571 TI - Myocardial protection during percutaneous transluminal coronary angioplasty. AB - Coronary angioplasty results in transient coronary artery occlusion. This article reviews with systemic and regional methods aimed at preventing the electrophysiologic and hemodynamic consequences of regional myocardial ischemia. PMID- 2977572 TI - Transcatheter therapy for congenital heart disease. AB - In summary, the cardiac catheterization suite has evolved from an invasive diagnostic facility into a primarily therapeutic laboratory. Through the use of balloons, blades, and plugs there are an increasing number of patients who may have definitive cardiac therapy on an outpatient basis, avoiding more invasive surgical interventions. PMID- 2977573 TI - Extra-weak chemiluminescence of drugs. IX. Extra-weak chemiluminescence of neocarzinostatin injection. PMID- 2977575 TI - In vivo PUVA and UVB sensitivity of various human epidermal Langerhans cell markers (ATPase, HLA-DR and T6)--dose-response and time-sequence studies. PMID- 2977574 TI - The topical treatment of onychomycosis using a new combined urea/imidazole preparation. PMID- 2977576 TI - Seborrhoeic keratoses and internal malignancies. A case control study. PMID- 2977577 TI - Onychomycosis due to Trichophyton soudanense. PMID- 2977578 TI - A comparison of doxycycline and minocycline in the treatment of acne vulgaris. PMID- 2977579 TI - Severe acne due to isoniazid. PMID- 2977580 TI - [Multipathway leading to platelet aggregation and in vivo platelet secretion in cerebral ischemia]. PMID- 2977581 TI - Rapid method for the isolation of bacteriophages from lysogens. AB - A rapid method for the isolation of bacteriophages from lysogens is described. Phages are induced in broth medium containing mitomycin C, which is then replicated onto agar medium. Molten medium containing indicator strains is then poured in these plates. Bacterial lysis is subsequently detected with tetrazolium containing broth. PMID- 2977582 TI - Replacement of the canine infrarenal aorta with an autogenous, living, tissue graft. PMID- 2977583 TI - Atrial natriuretic factor causes secretion in the isolated perfused rabbit ileum. PMID- 2977584 TI - [Reflex scoliosis in backache syndrome]. PMID- 2977585 TI - [Research on the relation of reninangiotensin-aldosterone and cardionatrin in endemic cretinism]. PMID- 2977586 TI - Influence of antihypertensive therapy on development and progression of diabetic glomerulopathy. AB - Capillary hypertension is suggested to be the underlying cause of microvascular disease affecting the kidney, the retina, and other organs and tissues in diabetic patients and animals. Hyperglycemia causes an expansion of extracellular volume, which induces a vasodilatory response. Hemodynamic adaptation to vasodilation leads to an increase in intracapillary hydraulic pressure, which subsequently causes vascular damage. In experimental animals, restoration of capillary pressure to normal levels by ingestion of a low-protein diet or administration of an angiotensin I-converting enzyme inhibitor has been shown to prevent microvascular damage in the kidney, and dietary protein restriction limits injury in the retina as well. Atrial natriuretic peptide, which is secreted by atrial myocytes in response to volume expansion, may be involved in mediation of the hemodynamic adaptation (vasodilatory response) that results in diabetic microvascular disease. PMID- 2977587 TI - The in vitro activity of ramoplanin (A-16686/MDL 62,198), vancomycin and teicoplanin against methicillin-susceptible and methicillin-resistant Staphylococcus spp. AB - Ramoplanin (A-16686/MDL 62,198) is a novel lipoglycopeptide antimicrobial, comprised of three closely related polypeptides containing chlorinated phenyl moieties and D-mannose, isolated from the fermentation products of Actinoplanes sp. ATCC 33076. The antimicrobial activity of ramoplanin is limited to Gram positive bacteria and its reportedly unacceptable administration side-effects suggest that any potential clinical role will be limited to the topical therapy of superficial skin infections and the eradication of bacteria, representing a possible nosocomial cross-infection source, from carriage sites. In this study the MICs of ramoplanin have been determined for methicillin-susceptible and methicillin-resistant isolates of Staphylococcus aureus, S. epidermidis and S. haemolyticus and compared with those of two glycopeptide antimicrobials, vancomycin and teicoplanin. MICs were determined using an agar incorporation technique in Mueller-Hinton medium with an inoculum of 10(5) cfu. Ramoplanin was 2-8 times more active than either vancomycin or teicoplanin against methicillin susceptible and methicillin-resistant isolates of S. aureus and methicillin susceptible isolates of S. epidermidis. Isolates of methicillin-resistant S. epidermidis and both methicillin-susceptible and -resistant isolates of S. haemolyticus were generally less susceptible to teicoplanin than to vancomycin. Ramoplanin was significantly more active than either vancomycin or teicoplanin against these isolates. These results suggest that the clinical evaluation of ramoplanin as a topical antibacterial agent for the control of superficial infections caused by Staphylococcus spp. and for the eradication of methicillin resistant S. aureus from carriage sites, is justified. PMID- 2977588 TI - Incidence and prognostic importance of silent ischaemia after PTCA: a prospective study. AB - To evaluate the influence of PTCA on symptomatic and asymptomatic ischaemic episodes in 94 patients, 24-h ambulatory, electrocardiographic Holter recordings were obtained before and after successful PTCA. Sixty-four per cent of patients had one-vessel disease, 28% two-vessel disease and 8% had three-vessel disease. Ischaemic episodes were present in 36% of patients before PTCA, of which 71% were silent; after PTCA, 23% of patients had ischaemic episodes, of which 98% were silent; thus silent ischaemic episodes were improved by PTCA to a lesser degree than symptomatic ischaemic episodes. Successful PTCA lead to a significant reduction in total number and duration of ischaemic episodes but not to a complete abolition. Patients with silent ischaemic episodes after PTCA had also a higher incidence of silent ischaemic episodes before PTCA. Functional and haemodynamic improvement was comparable in patients with and without silent ischaemic episodes. No specific cause for the persistent or newly appearing silent ischaemic episodes after PTCA could be identified; they are not indicative of an inadequate dilatation and cannot be considered as a risk factor for early restenosis. A possible explanation could be a traumatically induced, increased vascular tone in susceptible patients. PMID- 2977589 TI - [Ecologo-physiologic aspects of the occurrence of arterial hypertension in Leningrad (facts and hypotheses)]. AB - High rate of arterial hypertension occurrence in Leningrad and North-West of Europe is supposed to be associated with some ecological factors (soft water with low quantity of minerals) aiding the action of some main etiological factors of the hypertension. Some clinical-physiological approaches to solution of the problem are suggested. PMID- 2977590 TI - [Correlation of functional and structural changes of the cardiovascular system in hypertensive disease]. AB - Dynamics of left ventricular mass, minimal resistance of forearm arterial vessels and compliance of forearm venous vessels were studied in hypertensive patients during a 3-year effective antihypertensive treatment. Our findings suggest that pressure reduction and regression of myocardial hypertrophy develop in a parallel fashion while minimal resistance of vessels remains elevated due to the increasing contractile activity of smooth muscles. The presence of structural changes can only be assessed if the transmural pressure is the same along the whole arterial tree. PMID- 2977591 TI - [Several biophysical parameters of the cardiovascular system in patients with hypertensive disease]. AB - Biophysical assessment of circulation was performed in 135 patients with essential hypertension and 39 healthy subjects at rest and during exercise. An increase in arterial impedance was found to play a major part in the mechanisms of blood pressure elevation. In patients with eukinetic hemodynamics, the markedness of left ventricular hypertrophy was associated with adequate circulatory biophysical mode at rest and with the limitation of adaptive changes during exercise. In patients with hypokinetic hemodynamics, the hypertrophied myocardium adjusts itself to an increased after load at the expense of increments in the left ventricular oxygen demand (even at rest) and the exhaustion of myocardial functional reserve during muscular activity. PMID- 2977592 TI - The characterization of the protection by nicotinamide against the action of diabetogenic nitrosoureas. AB - The ability of various amide compounds to reverse the inhibitory effects of diabetogenic nitrosoureas on glucose stimulated insulin secretion was ascertained in isolated islets of Langerhans. Addition of nicotinamide 30 or more minutes after exposure of islets to streptozotocin (STZ) did not significantly reverse the insulinlytic effect of STZ. Not only nicotinamide isomers--isonicotinamide and picolinamide, but also other primary amides--pyrazinamide and benzamide were equally as effective as nicotinamide against nitrosourea and alloxan induced beta cytotoxicity. The protective ability of the amides was dose dependent in all cases and ranged from over 90% for 20 mM, 70% for 10 mM and 26% for 5 mM regardless of which isomer was used. The stronger the basicity of the ring nitrogen of pyridine derivative, the more potent the amide in protecting islets against the effect of either STZ or MNU. We have presented evidence to show that the protective ability of nicotinamide against diabetogenic nitrousoureas is not a specific property of nicotinamide but rather such action is shared by amides with stronger basic pyridine nucleus. PMID- 2977593 TI - Tolbutamide enhances insulin action on gluconeogenesis and on fructose 2,6 bisphosphate levels in isolated rat hepatocytes. AB - In hepatocytes isolated from fed rats and incubated either under basal conditions or in the presence of glucagon, tolbutamide reduced gluconeogenesis from (U-14C) pyruvate by increasing the cellular concentration of fructose 2,6-bisphosphate. Furthermore, this sulfonylurea enhanced the inhibitory action of insulin on glucagon-stimulated gluconeogenesis; this effect was accompanied by a more marked increase of the cellular concentration of fructose 2,6-bisphosphate than that elicited by either insulin or the sulfonylurea alone. In connection with this, tolbutamide--without significant modification of cellular cyclic AMP levels- raised the proportion of 6-phosphofructo 2-kinase in active form in hepatocytes incubated either under basal conditions or in the presence of glucagon, and reinforced the action of insulin in antagonizing the glucagon-mediated inactivation of this enzyme. PMID- 2977594 TI - [Complex psychophysiologic and biochemical study of the mechanisms of correcting a stable pathologic state of dependence on alcohol]. PMID- 2977595 TI - [Relationship between morphology of restenosis and clinical aspects in patients who had undergone coronary angioplasty]. AB - Although restenosis after percutaneous transluminal coronary angioplasty (PTCA) may have morphologic characteristics which are not similar to the original stenosis, it is unknown if a different morphology is also associated with a different clinical presentation. Eleven consecutive patients with angina and single vessel disease had a recurrence of symptoms and restenosis within 6 months of successful PTCA. Seven patients (group I) complained of spontaneous angina before PTCA and had a positive hyperventilation test, while 4 patients complained of exercise-induced angina and had a negative hyperventilation test (group II). In group I restenosis was associated with the same clinical presentation as before PTCA and the hyperventilation test was positive in 6 patients. The angiographic morphology of restenosis was changed in 4 patients. In group II restenosis occurred with the same clinical presentation as before PTCA, the hyperventilation test was negative in 3 patients while the angiographic morphology of restenosis was unchanged in only 1 patient. Although morphology of restenosis is different from the original stenosis, clinical presentation associated with restenosis is generally the same as before PTCA. The response to vasoactive stimuli is independent from the morphology of the lesion. PMID- 2977596 TI - [Coronary angioplasty in the aged]. AB - The safety and clinical efficacy of percutaneous transluminal coronary angioplasty (PTCA) in elderly patients has not been established. PTCA was attempted in 34 patients aged 65 or more (mean age 67.4). Patients were referred for angioplasty because of significant symptomatic ischemic heart disease with either stable, unstable angina or chest pain after myocardial infarction. Primary success was achieved in 29 patients (85.3%). Significant complications were encountered in three patients (8.8%): two emergency surgical procedures (5.8%), one transmural infarction (3%). In two patients (5.8%) the PTCA failed because the balloon dilating catheter didn't cross the tight stenosis. Follow-up data (mean 13.8 +/- 10.3 months) are available: 22 (73.4%) are asymptomatic. A clinically apparent recurrence occurred in 8 patients (26.6%). Coronary angiography was performed in 19 patients (63.3%) 6.3 +/- 2.4 months after PTCA. Angiographic restenosis occurred in 10 patients (52.6%). All patients with restenosis were ridilated with completely success. In conclusion, PTCA can be performed in elderly patients with a good success rate, an acceptable complication rate, a relatively low clinically apparent recurrence rate, and should be considered as a therapeutic modality for selected geriatric patients. PMID- 2977597 TI - [Usefulness of the modification of Pfannenstiel's incision by the Cherney method]. PMID- 2977598 TI - Red cell polymorphisms in Sardinia. AB - Acid phosphatase (ACP1), esterase D (ESD) and phosphoglucomutase 1 (PGM1) polymorphisms have been studied in Sardinia and the following gene frequencies have been found: ACP1*A = 0.235, ACP1*B = 0.684 and ACP1*C = 0.081; ESD*2 = 0.118 and PGM1*2 = 0.233. These findings confirm the genetic uniqueness of Sardinians compared to the other Italian and European populations. PMID- 2977599 TI - Immunoprophylaxis of hepatitis B in India. PMID- 2977600 TI - Robertsonian translocation with reciprocal product in the father of a 46, XY t(21q;21q) boy. PMID- 2977601 TI - Omphalopagus twins and twin transfusion syndrome. PMID- 2977602 TI - [A hepatitis patient comes for an extraction]. PMID- 2977603 TI - [Are there contraindications for the addition of noradrenaline to anesthetic solutions?]. PMID- 2977604 TI - [Oro-dental management of cancer patients]. PMID- 2977605 TI - Cell adhesion and phagocytosis promoted by monoclonal antibodies not directed against fibronectin receptors. AB - In this report we describe cell adhesion and phagocytosis promoted by two monoclonal antibodies that were selected for immunofluorescence staining of non permeabilized baby hamster kidney (BHK) cells. Anti-BHK1 staining was heaviest along cell margins, whereas anti-BHK2 staining was continuous along cell margins. Neither antibody stained elongated plaque structures such as were observed when cells were reacted with antibodies to fibronectin (FN) receptors. The monoclonal antibodies functioned as adhesion ligands in four different assays: attachment to culture dishes, spreading, binding of latex beads and phagocytosis. Anti-BHK1 and anti-BHK2 promoted attachment to culture dishes similarly, but anti-BHK2 was more effective at promoting cell spreading. Antibody-promoted cell spreading was inhibited by the peptides Ser-Asp-Gly-Arg and Gly-Arg-Gly-Asp-Ser-Pro but not by other, related, peptides tested. The monoclonal antibodies also promoted binding of latex beads, and the bead binding sites were motile, on the basis of their 'capping' response. Nevertheless, anti-BHK2 beads were phagocytosed by cells 5- to 20-fold more efficiently than anti-BHK1 beads. The binding sites for anti-BHK1 and anti-BHK2 were characterized by immunoprecipitation experiments. Anti-BHK1 binding sites contained 50K (K = 10(3) Mr) and 88K components under non-reducing conditions that migrated as a 51/53K doublet and a 93K component under reducing conditions. On the other hand, anti-BHK2 binding sites contained 88K and 110K components under non-reducing conditions that shifted to apparent 107K and 128K values when measured under reducing conditions. PMID- 2977606 TI - Mechanism of action of prothymosin alpha in the human autologous mixed lymphocyte response. AB - Prothymosin alpha(Prot alpha), an immunologically active polypeptide derived initially from rat thymus, and now pig thymus, was tested for its effect on autoantigen-induced human T cell proliferation in vitro. Pig ProT alpha was found to enhance the autologous mixed lymphocyte response (auto-MLR). Optimum enhancement was achieved at doses which varied among different donors. Treatment of the stimulatory monocytes with ProT alpha resulted in considerably higher auto MLR responses as compared to those with non treated monocytes. ProT alpha was without effect on T lymphocytes. In contrast, T lymphocytes exhibited enhanced proliferative activity when treated with ProT alpha in the environment of autologous monocytes. Moreover, supernatants from cultures of monocytes incubated with ProT alpha (ProT alpha-sup) were also shown to enhance the human auto-MLR either after addition in cultures or after preincubation with responder T lymphocytes. In addition, ProT alpha-sup did not demonstrate any detectable interleukin 1 (IL 1) or interleukin 2 (IL 2) - like activity. Furthermore, ProT alpha-sup induced an increase in IL 2 production in auto-MLR cultures. The enhancement of T-cell proliferation and IL 2 production by ProT alpha-sup was maximal when this material was added at the beginning of the auto-MLR, and no effect of ProT alpha-sup was seen if the latter was added 3 days after initiation of the culture. Finally, Prot alpha-sup was also shown to increase the expression of IL 2 receptors on T lymphocytes activated in the auto-MLR. These studies suggest that ProT alpha enhances the human auto-MLR through ProT alpha-sup which is released after interaction of monocytes with ProT alpha ProT alpha-sup then increases directly T lymphocyte proliferation by elevating IL 2 production and expression of IL 2 specific receptors on autoactivated T lymphocytes. PMID- 2977607 TI - Inhibition of graft-vs-host induced immunodeficiency with immunosuppressive therapy. AB - The pathologic features of the acute graft-vs-host disease occurring in unirradiated (C57Bl/6 X A/J)F1 mice injected intravenously with lymphocytes from the C57Bl/6 parent are similar to those reported for other parental----F1 hybrid combinations. When stimulated in culture with concanavalin A, lipopolysaccharide or alloantigen, spleen cells from B6AF1 mice that had been injected 11 days previously with B6 lymphocytes exhibited proliferative responses that were drastically reduced in comparison to the responses of spleen cells from F1 hosts injected with syngeneic lymphocytes. IL2 production in GVH spleen cell cultures was also diminished. Proliferative responses and IL2 production were partially restored in mice given immunosuppressive therapy with azathioprine, cyclosporin A or Sch 24937 a drug whose inhibitory effects on cellular and humoral immune responses in mice have recently been described. Phenotypic analyses by flow cytometry of the GVH splenocyte population indicated that the most consistent change in the GVH spleen was the appearance of an Lyt2+ L3T4+ T cell subset which in the majority of experiments was accompanied by an increase in cells expressing only the Lyt2 antigen. Both subpopulations were reduced in mice that had recovered immunological responsiveness following immunosuppressive therapy. The results suggest that in this GVH model the development of an immunodeficient state is directly related to the induction of an active T suppressor cell population and that such cells are effectively eliminated from the splenocyte population following treatment with some immunosuppressive drugs. PMID- 2977608 TI - Effect of BCG upon functional and phenotypic immune markers in rats bearing the Dunning R3327 MAT-LyLu prostatic adenocarcinoma. AB - Rats bearing (or not bearing) the Dunning R3327 MAT-LyLu prostatic adenocarcinoma were treated with Bacillus Calmette-Guerin (BCG) and evaluated for immune competence using functional and phenotypic markers. Tumor presence significantly depressed total T and helper T cell representation along with the helper/suppressor T cell ratio. Functional immunity, measured by phytohemmagglutinin (PHA) induced blastogenesis, was also significantly depressed. When BCG was administered to non-tumor bearing animals, it had no effect upon T cell subset distributions but significantly reduced PHA induced blastogenesis. BCG similarly administered to tumor bearing animals did not alter the depressed helper/suppressor T cell ratio found in tumor bearing rats, but did significantly elevate PHA induced blastogenesis. However, these elevated levels of functional immunity in BCG treated tumor-bearing rats remained significantly below normal. These data demonstrate a poor correlation between functional and phenotypic assessments of immune capability. PMID- 2977609 TI - [Disorders of the posterior pituitary]. PMID- 2977610 TI - [Urinary beta-thromboglobulin in preeclampsia]. PMID- 2977611 TI - [A case of a para bile duct lymphatic cyst with polycystic disease: a para bile duct lymphatic cyst treated with pure ethanol injection after ultrasonic drainage]. PMID- 2977612 TI - Molecular analysis of suppressor T cell receptors. PMID- 2977613 TI - Understanding suppressor cells: where have we gone wrong? PMID- 2977614 TI - Utilization of digital image processing to study dynein arms (ATPase) in normal and immotile cilia. AB - Tracheal ciliary cross sections were examined with scanning transmission electron microscopy and the resultant images were digitized for image enhancement. A gray scale histogram of each ciliary image was produced and manipulated to enhance the image for dynein arms. Tracheal epithelial tissue from the pig, rabbit, and dog, including dogs with immotile cilia syndrome, was examined by using this technique. Tissue from each animal was fixed with each of three different fixatives and sections were evaluated for preservation of dynein arms. The same fixative did not consistently provide optimal fixation for ciliary dynein arms in all three species examined. Each species, therefore, must be evaluated to determine the optimal fixative for preservation of normal ciliary ultrastructure. Digital image processing provides a mechanism for enhancing dynein arms in situ without the need for addition of special stains or the use of techniques such as image summation. With this technique it has been shown that about two-thirds of outer dynein arms are partially or completely missing on cilia from dogs with immotile cilia syndrome. PMID- 2977615 TI - High contrast fluorescence imaging using two-wavelength laser excitation and image processing. PMID- 2977616 TI - Photodynamic effects of hematoporphyrin-derivative on enzyme activities of murine L929 fibroblasts. AB - Photodynamic treatment of murine L929 fibroblasts with hematoporphyrin-derivative resulted in the inactivation of cytosolic, mitochondrial and lysosomal enzymes and in a decrease in cellular adenosine triphosphate and reduced glutathione concentrations. Comparison of these results with those of previous studies revealed that transmembrane transport systems and DNA repair enzymes are inactivated after much shorter illumination periods than are intracellular enzymes. Although the pattern of photodynamic damage altered by varying the protocol of preincubation with hematoporphyrin-derivative and washing, it appeared that under all experimental conditions the plasma membrane was much more sensitive to photodynamic damage than were the intracellular enzymes. Lysosomal membrane disruption with subsequent detrimental release of lysosomal enzymes has been implicated previously in certain forms of porphyrin-induced photodynamic cell destruction. Cytochemical studies on enzyme localization virtually exclude such a mechanism in hematoporphyrin-derivative-induced cell inactivation in L929 fibroblasts. PMID- 2977617 TI - Porphyrin derivatives having physical and chemical characteristics similar to those of the active components of hematoporphyrin derivative and with very strong photosensitizing effects. AB - The tumour-localizing fraction of hematoporphyrin derivative (Hpd) is thought to possess an essentially diporphyrin ether structure or, alternatively, a diporphyrin ester structure, the properties of which facilitate its retention in malignant cells and its biological activity on irradiation. To elucidate this problem further, we have synthesized the dimethyl, diethyl, dipropyl, di-n-butyl and di-iso-butyl ethers of hematoporphyrin. These ethers show chromatographic properties very similar to those of the active components of Hpd. Furthermore, they are much better photosensitizers in a cellular system than are crude Hpd or Photofrin II, and, like the components of Hpd, they are taken up and retained by cells according to their degree of non-polarity. PMID- 2977618 TI - Hematoporphyrin derivative photosensitization and cellular fluorescent photoproduct formation. AB - A new fluorescence band with an emission maximum at 460 nm was observed in S180 tumour cells and human cancer tissues photosensitized by Y-HPD. This fluorescence was the result of a photochemical reaction involving specific proteins and Y-HPD in the cells. PMID- 2977619 TI - A culture medium for Paracoccidioides brasiliensis with high plating efficiency, and the effect of siderophores. AB - The plating efficiency of Paracoccidioides brasiliensis on standard mycological media is poor, impairing its isolation and recovery from various sources, particularly infected tissues. We describe a medium that markedly improves P. brasiliensis plating efficiency. It consists of a synthetic medium (modified McVeigh-Morton) supplemented with 4% (v:v) horse serum and 5% (v:v) culture filtrate from stationary phase P. brasiliensis cultures. A commercially available medium (brain-heart infusion), ordinarily inferior to unsupplemented McVeigh Morton medium, is at least as efficacious as supplemented McVeigh-Morton medium when supplemented in this manner. We show that plating efficiency varies among P. brasiliensis isolates and can even vary with the isolate's history of passage in culture. In contrast, all isolates studied could produce the growth enhancing factors present in culture filtrate. Some siderophores produced by other fungi can be substituted for the culture filtrate, whereas others can be substituted for both the filtrate and serum. The enhancing effect of filtrate and/or serum could be removed by chelating iron. P. brasiliensis-produced siderophores are likely to be the growth enhancing moiety in culture filtrates. PMID- 2977620 TI - [Studies on metabolism and disposition of sizofiran (SPG), an anti-tumor polysaccharide. I. Excretion and tissue distribution of 14C-SPG]. PMID- 2977621 TI - [Unilateral isthmic lysis in x-ray computed tomography]. AB - Condensing pediculolaminar hypertrophy of mechanical origin is known as "Dahu's sign". Dahu's sign corresponds to hypertrophy of the vertebral hemi-arch found by CT scan and leads to suspicion of controlateral lysis of the isthmus. It is the CT scan equivalent of vertebral "anisocorism" by conventional radiology. PMID- 2977622 TI - Cellular immunity and suppressor T cell function in asthmatic children on prolonged ketotifen therapy. AB - This study was undertaken to investigate the effect of ketotifen on the immune system of asthmatic children receiving prophylactic treatment with ketotifen. E. rosettes, mitogenic response to phytohemagglutinin (PHA) and concanavalin A (Con A) as well as Con A induced suppression were measured in 15 asthmatic patients before and 1-3 months after treatment with ketotifen (1 mg x 2/day), and in 20 normal healthy controls. No significant difference in immunological parameters studied were found between the pre and post treatment periods in the asthmatic patients, neither was any difference demonstrated between the asthmatic and control group studied. PMID- 2977623 TI - NK cell activity and monocyte dysfunctions in a patient with common variable hypogammaglobulinemia. AB - A 57-year-old man with a history of recurrent infections from the age of 50 was hospitalized with a diagnosis of common variable hypogammaglobulinemia (CVH). Immunological studies revealed a severe reduction of circulating immunoglobulins of all classes. Immunophenotyping of peripheral blood mononuclear cells (PBMC) with monoclonal antibodies, revealed normal values of total B and T cells with CD4/CD8 ratio sharply reduced (0.35) as compared to normal (1.6) because of an increase of CD8 and a decrease of CD4 cells. The surface expression of IL-2 receptor was normal. Natural cytotoxic and phagocytic system presented several abnormalities: a deep impairment of NK activity was found in spite of a normal number of NK cells, as ascertained by Leu 19 and B73.1 monoclonal antibodies. The defective NK activity was not restored by interferon alfa, but was normalized by recombinant IL-2. Phagocytic function, as defined by zymosan-stimulated O2- production was almost absent. The involvement of natural cytotoxic and phagocytic systems in CVH has been rarely reported; the possible causative role of a chronic viral infection (Epstein-Barr virus?) is discussed, on the base of anamnesis. PMID- 2977624 TI - [Increased complexes between protease and protease inhibitor in the plasma from patients with neoplasm]. PMID- 2977625 TI - The regulation of natural killer cell activity by splenic nonspecific suppressor cells and its modification in cancer patients. AB - Spleen cells (SC), splenic venous blood lymphocytes (SVL) and peripheral blood lymphocytes (PBL) from gastric and esophageal cancer patients were simultaneously tested for natural killer (NK) and nonspecific suppressor (Ts) cell activities. Furthermore, the influence of Ts activity on the augmentation of NK activity by a biological response modifier (BRM) was also investigated. Positive Ts activities were frequently detected in the SC, SVL and PBL of advanced cancer patients. The NK activities of SC and SVL were maintained even in advanced cancer patients, though significantly depressed NK activities were observed in the PBL of advanced cases. Cancer patient SC, SVL and PBL with positive Ts activity showed low NK activities. Moreover, the NK activities of SVL and PBL were low in the patients with positive Ts activity in SC. The NK activity of normal control PBL was strongly augmented by interleukin 2, interferon and OK-432. These BRMs exhibited comparable capacities to augment the NK activities of SC, SVL and PBL with negative Ts activity in cancer patients, however, the effects of these agents seemed to be low in cells with a positive Ts activity. These results suggested that NK activity might be regulated by nonspecific suppressor cells and the presence of suppressor cells might affect the augmentation of NK activity through BRM in circulating blood lymphocytes and also in spleen cells. PMID- 2977627 TI - [Surgical treatment of congenital cardiac defects in a patient with Cantrell syndrome; a case report]. PMID- 2977626 TI - The antiproliferative effects of fluoropyrimidine derivatives against human tumor xenografts in a subrenal capsule assay. AB - The antiproliferative effects of the fluoropyrimidine derivatives, 5-fluorouracil (5-FU), 1-(2-tetrahydrofuryl)-5-fluorouracil (Tegafur), UFT, 1-hexylcarbamoyl-5 fluorouracil (HCFU), and 5'-deoxy-5-fluorouracil (5'DFUR), were investigated in a 4 day subrenal capsule assay. The antiproliferative effects against two human tumor xenografts established in athymic mice were examined after treatment with three different doses of each anticancer agent, and the adequate dose of each anticancer agent in this experimental system was estimated as: 473 mg/kg for Tegafur, 433 mg/kg for UFT, 50 mg/kg for HCFU and 185 mg/kg for 5'DFUR, respectively. A comparative study of the antiproliferative effects of fluoropyrimidine derivatives was carried out against 7 xenografts. According to our criteria of positive tumor response, the effective rates were: 1 of 7 (14.3 per cent) by 5-FU, 2 of 7 (28.6 per cent) by Tegafur, 2 of 7 (28.6 per cent) by UFT, 1 of 6 (16.7 per cent) by HCFU, and 1 of 4 (25.0 per cent) by 5'DFUR, respectively. Although no statistical differences were demonstrated between the agents, the utility of a chemosensitivity test before clinical use was suggested. PMID- 2977628 TI - [A case of a giant postoperative hernia successfully treated with Bulgarian antimicrobial polyamide mesh]. PMID- 2977629 TI - Treatment of lower back pain with "pressing and pulling" at Kunlun (UB 60) method. PMID- 2977630 TI - Expression of the Escherichia coli lamB gene in Vibrio cholerae. AB - A phage lambda mediated transduction system was devised to facilitate molecular analysis of Vibrio cholerae. A lamB expression plasmid, pAMH62 was introduced into Vibrio cholerae by conjugation. The resulting V. cholerae derivatives harboring pAMH62 produced substantial amounts of the LamB protein. This protein was properly inserted into the outer membrane, as suggested by (i) its localization into the cell envelope, (ii) its association with the peptidoglycan layer of the cell wall, and (iii) its function as receptor for phage lambda. In vivo packaged cosmids were efficiently transduced into these strains of V. cholerae. PMID- 2977631 TI - [Trunk and abdominal wall edema in dermatomyositis]. PMID- 2977632 TI - [Pathogenetic mechanism of cardiovascular changes in carbon monoxide exposure]. PMID- 2977633 TI - [Allergic contact dermatitis caused by rubber. Evidence of a new group sensitization to p-phenylenediamine derivatives]. PMID- 2977634 TI - [Dental abnormalities in several malformation conditions (cleft lip and/or palate, multiple malformation syndromes) in several dysostosis in children]. PMID- 2977635 TI - [Current approaches in the treatment of gastroschisis]. PMID- 2977636 TI - [Change in immunologic reactivity in children with penetrating eye injuries]. PMID- 2977637 TI - [Results and prospects of the development of traumatologic-orthopedic services in the republics of Central Asia and Kazakhstan]. PMID- 2977638 TI - [A method of saturation of tissues of the limbs with morphocycline]. PMID- 2977639 TI - [Multiple injuries among sailors on ships]. PMID- 2977640 TI - [Keratoconjunctivitis sicca during treatment with 13-cis-retinoic acid (isotretinoin)]. PMID- 2977641 TI - Human model for studying in vitro the regulating function of specific and scavenger LDL receptors. AB - Measurements of the specific and scavenger LDL receptor activities in different disorders do not reveal all the causes and origin of hypercholesterolaemia. Studying the failure of the intracellular regulation of these receptors can give us some new information about the background of atherosclerosis. Therefore, we have tried to develop an in vitro model which is suitable for studying the specific and scavenger LDL receptor activities and their most important regulating functions, i.e. the inhibition of HMG-Co-A reductase by native LDL and the apo E secretion induced by acLDL. MDMs cultured for three days have turned out to provide a good model to test these functions. PMID- 2977642 TI - Changes in vasopressin-converting aminopeptidase activity in the rat pineal gland during summer: relationship to vasopressin contents. AB - Vasopressin (VP)-converting aminopeptidase (VP-AP) activity and VP contents were measured in single rat pineal glands during the summer of two successive years. The peptidase activity decreased significantly in August. The lowest activity (+/ SEM) of 0.18 +/- 0.02 pmol.hour-1 was recorded on August 14, compared to the basal activity of 0.25 +/- 0.01 pmol.hour-1 in July and September of 1986. The change with similar percentage occurred in the same period of 1987. The specific activity of the enzyme in the crude homogenate, 15,000 g pellet and supernatant fraction of rat pineal glands, exhibited the same pattern of variations. The decrease in peptidase activity coincided with the previously reported dramatic rise in pineal VP content in early August which was confirmed in this series of experiments. Another peptidase, the so-called gamma-endorphin generating endopeptidase (gamma-EGE) activity, and beta-endorphin-related peptides in the pineal gland did not change in this period. The results show that the variations of pineal VP contents and VP-AP activity during summer are not general for other peptides and peptidases. The coincidence of opposite changes in VP content and VP AP activity of the pineal gland may indicate a role of the peptidase activity to regulate the VP content. PMID- 2977643 TI - Specificity of serine proteases for cleavage sites on proatrial natriuretic factor. AB - An immunological approach was used to investigate the specificity of protease cleavage sites on proANF. Cleavage of 35S-cysteine biosynthetically-labeled proANF by whole serum, thrombin and kallikrein was examined. Reaction products were immunoprecipitated with two antibodies directed to different epitopes: a previously characterized antibody directed toward the carboxy-terminus of ANF103 126, which cross reacts with proANF, ANF99-126 and ANF103-126, and a newly prepared antisera to synthetic ANF99-105, which uniquely recognizes ANF99-126, but not proANF or ANF103-126. With increasing time of incubation with rat serum, proANF is sequentially cleaved at the C-terminus of a monobasic Pro-Arg dipeptide sequence to form ANF99-126, and then at the C-terminus of a dibasic Arg-Arg dipeptide sequence to yield ANF103-126. This cleavage activity of serum is blocked by leupeptin (40 micrograms/ml), but not by hirudin (100 nM), a specific inhibitor of thrombin, or by aprotinin (200 KIU/ml), a kallikrein inhibitor. When 100-fold purified serum cleavage enzyme was used in place of crude serum, similar results were obtained. Thrombin cleaves proANF only at the monobasic site to produce ANF99-126 while kallikrein cleaves only at the dibasic site to produce ANF103-126. As expected, the generation of these cleavage products can be inhibited by hirudin or aprotinin respectively. These data indicate that the substrate specificity of the serum cleavage activity is broader than that of thrombin or kallikrein, and that cleavage of proANF by serum proteases may be influenced by conformational restraints. The methods developed here should help in the future characterization of the physiological proANF cleaving enzyme. PMID- 2977644 TI - Interaction of putative vasopressin receptors in rat brain and bovine pituitary gland with a vasopressin anti-idiotype antibody as revealed by immunoblotting. AB - Vasopressin (AVP)-binding proteins were obtained from rat brain and the anterior and posterior lobes of bovine pituitary glands by (a) preparation of crude membranes, (b) solubilization of membrane proteins, (c) passage through an affinity column containing immobilized AVP, and (d) elution from the column with excess AVP. Gel electrophoresis revealed protein bands of 55 and 62 kilodaltons in rat brain, bovine posterior lobe and, to a far lesser extent, in the anterior lobe, which were similar to those previously identified in rat brain to be associated with AVP binding. Immunoblotting demonstrated that the 55 kilodalton bands of rat brain and bovine pituitary gland were selectively immunoreactive with an AVP anti-idiotype antibody. In addition, immunoreactivity occurred with a 62 kilodalton component of rat brain. PMID- 2977645 TI - [Presence of immune complexes in patients with peptic ulcer]. PMID- 2977646 TI - Prostaglandin D2 inhibits pentylenetetrazole-induced convulsions in rats by a serotonin-mediated mechanism. AB - Prostaglandins (PGs) of the E series are known to exert anticonvulsant action in experimental animals. Earlier studies from this laboratory have indicated that PGE1 inhibits pentylenetetrazole (PTZ)-induced convulsions in rats through a serotonin-mediated mechanism. PGD2, the major PG in the rodent brain, shares a number of central pharmacological actions of the PGEs, and like the latter it potentiates the anticonvulsant action of phenobarbitone and phenytoin in rats. The present study was undertaken to investigate the putative anticonvulsant action of PGD2 against PTZ-induced convulsions in rats and to evaluate the role of serotonin in the anticonvulsant action of PGD2. PGD2 (5, 10, and 20 micrograms, icv) produced a dose-related inhibition of PTZ-induced clonic convulsions in rats. The anticonvulsant action of PGD2 (20 micrograms, icv) was significantly attenuated following pretreatment of the rats with pharmacologic agents known to reduce central serotonergic activity, including 5,6 dihydroxytryptamine, a selective neurotoxin for serotonergic neurons, p chlorophenylalanine, a specific inhibitor of serotonin biosynthesis, metergoline, a serotonin postsynaptic receptor antagonist, and quipazine, which is known to inhibit neuronal release of serotonin. These findings, in conjunction with an earlier study from this laboratory indicating that PGD2 augments rat brain serotonergic activity, suggest that the anticonvulsant activity of PGD2 against PTZ-induced convulsions in rats is mediated through a serotonergic mechanism. PMID- 2977647 TI - Inhibition of intestinal pyrimidine nucleoside phosphorylases. AB - The activity of 5'-deoxy-5-fluorouridine (dFUR) depends on its activation to 5 fluorouracil (FU) by pyrimidine nucleoside phosphorylases. These enzymes are found in tumors and normal tissues, with the highest activity in the small intestines. The present study examined the inhibition of dFUR phosphorolysis in intestinal tissues. dFUR metabolism in intestinal homogenates was inhibited by uracil (U), uridine (UR), and thymidine (TdR), which are the normal substrates for the phosphorylases. Conversely dFUR reduced the metabolism of these inhibitors. A good agreement was found between the observed data and the computer fitted data using the equations for competitive inhibition between dFUR and the inhibitors. In the absence of inhibitors, the Vmax of dFUR phosphorolysis was 47.1 +/- 4.9 microM/min and the apparent Km was 910 +/- 167 microM. The Vmax was unaltered by the inhibitors, while the Km was increased with increasing inhibitor concentrations. The maximal inhibition of dFUR metabolism by UR and TdR was about 80%. The Ki's were 372 microM for U, 87.2 microM for UR, and 112 microM for TdR and are orders of magnitude higher than their reported endogenous serum concentrations. The rate of dFUR phosphorolysis to FU in the intact intestinal epithelial crypt cells, indicated by the ratio of FU to dFUR in the intracellular fluid, was reduced by UR in a concentration-dependent fashion. These data indicate that the naturally occurring pyrimidines inhibit competitively the dFUR metabolism by the intestinal phosphorylases, that this inhibition occurs at concentrations much higher than the circulating endogenous levels, and that phosphorolysis is the major route of dFUR metabolism. PMID- 2977648 TI - Nonenzymatic and enzymatic hydrolysis of 5-fluoro-2'-deoxyuridine (FUdR) esters. AB - The chemical and enzymatic reactivity of 5-fluoro-2'-deoxyuridine prodrugs esterified at the 3' and 5' positions with several acyl groups has been investigated. The enzymatic reactivity was affected by the acyl structure, the site of esterification, and the number of esters in the prodrug molecule. PMID- 2977649 TI - A biotin-avidin-based enzyme immunoassay for beta h-endorphin. AB - An avidin-biotin enzyme-linked immunosorbent assay (ELISA) is described for beta h-endorphin (beta h-EP). Microtiter plates coated with commercially available antibodies were used together with beta h-EP tracer derivatives that were biotinylated in positions 24, 28, and 29 via a C6 spacer arm. Nonspecific binding of biotinylated derivatives to the microtiter plates was blocked with a mixture of 1% casein and 10% ethanolamine in 0.1 M NaHCO3. A sequential saturation procedure using a high-affinity antiserum in combination with an avidin-alkaline phosphatase complex matched the sensitivity of reported radioimmunoassays (RIAs), with a detection limit of 0.5 fmol/assay. The intra- and interassay coefficients of variation were 5 and 12%, respectively. Results obtained by ELISA and RIA showed good correlations (r = 0.95). The beta-EP concentration in extracted rat plasma after high-performance liquid chromatographic (HPLC) fractionation was determined by this method to be 1600 fmol/mol. PMID- 2977650 TI - [EDLS (endogenous digitalis-like substance(s))--detection, chemistry, and physiologic function]. AB - Human beings and higher animals contain compounds which interact with the Na+/K+ ATPase of the heart muscle and other organs like the cardiac glycosides, and bind to cardiac glycoside-specific antibodies [endogenous digitalis-like substance(s), EDLS]. EDLS cause increased natriuresis. The level of EDLS of the blood is raised under physiological stress situations (e.g., pregnancy and delivery and at certain pathophysiological conditions (e.g., hypertony). The EDLS are low molecular compounds. As yet their chemical structure is unknown. PMID- 2977651 TI - [Natriuretic peptide family: ANP and BNP]. PMID- 2977652 TI - [Biosynthesis and secretion of atrial natriuretic peptide]. PMID- 2977653 TI - [Role of ANP in the regulation of blood pressure]. PMID- 2977654 TI - [Lp(a)--the current state of knowledge and clinical significance]. PMID- 2977656 TI - Localization and properties of the ATPase in the inner ear of rat. PMID- 2977655 TI - Improved tolerance to ischemia in hypertrophied myocardium pre and post cardiopulmonary bypass use of a calcium-channel blocker (diltiazem). PMID- 2977657 TI - [Analysis of the clinical and angiographic parameters related to restenosis after angioplasty]. PMID- 2977659 TI - [Correlations between serum ANF concentration and cardiac functional indices assessed by two-dimensional echocardiography and pulse Doppler method in various cardiac diseases]. PMID- 2977658 TI - [Decrease of HCG level after conservative laparoscopic treatment of extra-uterine pregnancy]. AB - HCG levels have been studied in 76 patients following laparoscopic treatment using laparotomy. This shows that early detection of treatment failure is possible of ectopic pregnancy. The decrease of HCG was the same as after conservative treatment from the third or fifth post-operative day onwards. The authors report a scheme for the post-operative follow-up after laparoscopic treatment of ectopic pregnancy based on the rate of HCG decrease. PMID- 2977660 TI - [Characteristics of immunologic indices in patients with rheumatoid arthritis]. PMID- 2977662 TI - [Proceedings of the 33d meeting of the Lombard Urologic Society. Milan, 11 June 1988. Problems of urologic surgery in the irradiated pelvis]. PMID- 2977661 TI - [Huntington chorea in the province of Ferrara from 1971 to 1987. Descriptive study]. AB - In the context of a multidisciplinary study program whose purpose is to investigate the genetic aspects of Huntington's Chorea (HC), the authors conducted an epidemiological descriptive research extended to the population residing in the province of Ferrara in a time period including the years from 1971 to 1987. On December 31st, 1987 we estimated a prevalence rate of 1.89 cases of HC per 100,000 inhabitants; in the years 1971-1987 the incidence rate was of 0.11 per 100,000/year and the mortality rate of 0.06 per 100,000/year. In the last years considered for the study, the incidence and the prevalence showed a relative increase indicating that HC still exists in the ferrarese population, despite a greater public awareness. With the aim to organize preventive actions and to reach the preclinical diagnosis of the disease we singled out 83 subjects at risk the study area. PMID- 2977663 TI - [Radiobiology of injuries of the pelvic structures]. PMID- 2977665 TI - [Treatment of cystitis caused by radiation]. PMID- 2977664 TI - [Functional bladder-urethral injuries in the irradiated pelvis]. PMID- 2977666 TI - [Salvage cystectomy]. PMID- 2977667 TI - [Problems in surgical repair of ureteral injuries in the irradiated pelvis]. PMID- 2977668 TI - ["Terrorist" and optimist for the handicapped. Interview by Lars Peter Bergqvist]. PMID- 2977669 TI - [Disorders of the heart rhythm in patients with chronic obstructive lung diseases]. PMID- 2977670 TI - [The initial results of the sanatorium rehabilitation of patients having had an acute myocardial infarct in a mid-alpine climate]. AB - Short-term results of rehabilitation of 1046 CHD patients at a cardiological sanatorium in mountains at average altitude (1600 m above the sea level) were studied. Large focal myocardial infarction (MI) was found in 738 of them, small focal MI in 249, and acute focal myocardial dystrophy of ischemic genesis in 59. Of 1046 patients considerable improvement of the status after rehabilitation therapy at the sanatorium was noted in 27.6%, improvement in 64.5%, no changes in 6.1%, deterioration in 1.7%; one patient died. This type of therapy in a mountainous climate at average altitude yielded good immediate results in most of the patients (92.06%) with MI and could be recommended for rehabilitation of CHD patients at the 2nd stage of treatment at a sanatorium. PMID- 2977671 TI - [The role of suppressor cells in congenital influenza infection in mice]. AB - Young mice with congenital influenza infection have lower immune responsiveness of lymphocytes to nonspecific mitogens and influenza virus antigens. Lymphocytes of such animals inhibit proliferation of normal lymphoid cells activated with concanavalin A and immune lymphocytes activated with influenza virus antigens. It is assumed that in congenital influenza infection one of the possible mechanisms of immunosuppression in mice is the activation of suppressor T-cells. PMID- 2977672 TI - [Detection of antibodies using recombinant strains of influenza viruses in the sera of various species of birds circulating in the territories of the Ukrainian and Azerbaijan SSRs]. AB - Examinations of blood sera from different species of birds trapped in the Ukrainian and Azerbaijan SSRs using diagnostic preparations from the influenza A/sea gull/Maryland/704/77 virus strain and a recombinant R117 derived from it revealed the presence of antibodies to hemagglutinin H13. The diagnostic preparation produced from the recombinant strain was found to be more active in the detection of antibodies in avian sera. PMID- 2977673 TI - [Conservative complex treatment of lumbar vertebral syndrome in inpatient conditions]. PMID- 2977674 TI - Defective myo-inositol-1-phosphate synthase production in an inositolless double mutant Neurospora crassa strain. AB - A slow growing inl+/- mutant was isolated from an inositol dependent (inl) Neurospora crassa strain. The latter strain produces defective myo-inositol-1 phosphate synthase which has residual activity. Inositol, similarly to that found in wild and inl mutant strains, represses the enzyme production in the inl+/- strain as well. Withdrawing inositol from the medium results in derepression of the enzyme synthesis. Derepression is hindered by cycloheximide. Inl+/- character in the double mutant is brought about by overproduction of the defective myo inositol-1-phosphate synthase. PMID- 2977675 TI - Structure and expression of cDNA for calphobindin. PMID- 2977676 TI - [Chronic streptococcal tonsillitis: significance of specific blastogenic response test and possibilities of its clinical applications]. PMID- 2977677 TI - [Effect of glucomacropeptide on the cellular interrelations of the humoral immune response]. PMID- 2977678 TI - Mast cells in experimental and human cancerogenesis. PMID- 2977679 TI - Sinsemilla and its cultivation in climatic conditions of Czechoslovakia--a potential danger of abuse of marihuana of high quality. PMID- 2977680 TI - Evaluation of the diagnostical significance of lectin histochemistry in breast malignant and benign lesions. PMID- 2977681 TI - Recognizing lung pseudolymphoma (casuistic case). PMID- 2977682 TI - Effect of corticoid treatment upon the measles virus-induced leukocyte adherence inhibition. PMID- 2977683 TI - Trigeminofacial reflex. III. Effect of auriculotherapy. PMID- 2977684 TI - Metabolism of magnesium and zinc in skeletal system. PMID- 2977686 TI - Effect of genetic heterogeneity of hypertriglyceridemia upon the risk of coronary heart disease. PMID- 2977685 TI - Systemic and local factors in controlling of bone metabolism. PMID- 2977687 TI - [The prognostic significance of an ST-segment depression during the bicycle ergometric examination of men]. PMID- 2977688 TI - Membrane plasmapheresis and its current possibilities in internal medicine. PMID- 2977689 TI - Bone marrow necrosis (a report on five intravitally recognized cases and a brief review of the literature). PMID- 2977690 TI - Problems of the evaluation of occupational vasoneurosis in medical practice--a case report. PMID- 2977691 TI - Trichological alterations in women during pregnancy and after delivery. PMID- 2977692 TI - Changes in colour temperature characteristic of cholesteric temperature indicators provoked by UV radiation. PMID- 2977693 TI - Knee arthrography. PMID- 2977694 TI - Investigation of the behavior of enuretic children in stomatological surgery. PMID- 2977695 TI - Mechanical preparation and cleansing of root canals in vitro. PMID- 2977696 TI - [Morphometry of the dental tissues of human teeth in the primary and secondary dentitions]. PMID- 2977698 TI - Parameter estimation in the logistic response model. PMID- 2977697 TI - [Use of holographic interferometry and the moire method in fields of medicine]. PMID- 2977699 TI - Quantitative morphometric evaluation of changes in the small intestine mucosa of laboratory rats after X-ray and gamma irradiation. PMID- 2977700 TI - Empirical Bayesian estimation in the logistic model. PMID- 2977701 TI - Fisher information in the logistic model. PMID- 2977702 TI - Causes, prevention and therapy of premature labour. PMID- 2977703 TI - [Use of the infinitive as an adverbial clause in English technical medical texts]. PMID- 2977704 TI - [The role of the lymph nodes in the generalization of the process in experimental apical periodontitis]. PMID- 2977705 TI - Primary tumours of the heart. PMID- 2977706 TI - Semiautomatic operation of the rotatory microtome II. PMID- 2977707 TI - Prof. MUDr. Jan Hrbek, DrSc. The sixty-fifth anniversary. PMID- 2977708 TI - [On the birthday of Prof. Zdenek Sery (born 23 September 1917)]. PMID- 2977709 TI - Twenty-five years of the Second Department of Surgery of the University Hospital Olomouc. PMID- 2977710 TI - Effect of morphine on leukocyte kinetics in rabbits. PMID- 2977711 TI - Histochemical study of reaction of the nucleus supraopticus of rat brain to irradiation with 500 Gy. PMID- 2977712 TI - Histochemical changes of capillaries in rat brain cortex after irradiation with supralethal doses of gamma-radiation. PMID- 2977713 TI - Cellular reaction to concanavalin A studied by the skin window method in healthy subjects and in patients with malignant tumors. PMID- 2977714 TI - Local immune response in laryngeal cancer: a prognostic view. PMID- 2977715 TI - Tumor immunity in oral cancer: a clinico-pathological study. PMID- 2977716 TI - The role of thymidylate synthase in the response to fluoropyrimidine-folinic acid combinations. AB - A panel of human colorectal tumor cell lines has been examined to determine the role of TS in the response to fluoropyrimidine antimetabolites. Among these cell lines, the response to FdUrd does not correlate with the levels of TS. In cell lines HCT 116 and RCA, which are poorly responsive to FdUrd, structural alterations in TS have been identified. In HCT 116, two TS polypeptides are present: a common form, occurring in all the cell lines and a variant form. The variant TS polypeptide has a reduced affinity for the TS ligands, FdUMP and CH2H4PteGlu, relative to the common TS polypeptide. Clonal populations of HCT 116 that overproduce each form have been isolated. Clones that overproduce the variant polypeptide are 4-fold less responsive to TS-directed cytotoxic agents than those that overproduce the common; thus, the presence of the variant TS is associated with a reduced response to TS-directed cytotoxic agents. The response of cell line RCA to FdUrd is dependent upon the extracellular CF concentration: response increases as CF is increased. RCA contains a TS enzyme with reduced affinity for CH2H4PteGlu, relative to cell line C, which is sensitive to FdUrd at all CF concentrations. Both cells form high chain-length polyglutamates of CH2H4PteGlu at CF concentrations in which the response to FdUrd differs by 4 fold. In RCA, the TS structural gene is variant, relative to the other cell lines. This variation may underlie the altered enzyme affinity for CH2H4PteGlu and the sensitivity to modulation of FdUrd response by CF. PMID- 2977717 TI - Tumor cell responses to inhibition of thymidylate synthase. AB - Whether inhibition of thymidylate synthase is lethal to a population of tumor cells depends upon three factors: 1) the dependence of the cells upon de novo synthesis of thymidine nucleotides; 2) the length of time enzyme is inhibited and the requirement for thymidine nucleotides during this period; and 3) the biochemical responses of the cells to the initial inhibition of enzyme, many of which interfere with maintenance of thymidylate synthase in an inhibited state. Following inhibition of thymidylate synthase, deoxyuridylate accumulates, as does the cellular content of thymidylate synthase. In addition, the initially formed enzyme-inhibitor complexes dissociate. These biochemical sequelae alter the effectiveness of the blockade of thymidylate synthase in a time-dependent, continuously-changing manner. Whether cell kill occurs depends on whether the dynamic balance of these factors allows a sufficiently low enzymatic activity to be maintained for a long enough period of time. An analysis of this interaction of factors leads us to the conclusions that efficient tumor cell kill with fluoropyrimidines is best attained by combination with reduced folate cofactors and inhibitors of deoxypyrimidine biosynthesis. Each of these agents modifies the response of tumor cells with the result that the fluorodeoxyuridylate-induced inhibition of thymidylate synthase is maintained. This analysis also suggests that folate analogs inhibitory to thymidylate synthase are more compatible than pyrimidine analogs with inhibition of thymidylate synthase as an approach to cancer chemotherapy. PMID- 2977718 TI - Immune response to vaccinia virus and recombinant virus products in dogs. AB - A study was undertaken to determine the safety and suitability of vaccinia virus as an eukaryotic expression vector in dogs. Clinical signs were not seen in inoculated dogs, with the exception of small nodules at the site of inoculation. Vaccinia virus did not spread from dogs that were inoculated by SC (n = 5), intradermal (ID; n = 13), or intranasal (n = 3) routes to noninoculated dogs maintained in close contact. Replication of vaccinia virus appeared to be restricted in dogs because greater than or equal to 10(5) plaque-forming units of virus were required to induce an immune response by ID inoculation. Results were better with ID inoculation than with SC or intranasal inoculations. Repeated inoculations enhanced serum antibody titers, and annual reinoculation resulted in boosting of antibody titers. Maternal antibody interfered with virus replication and antibody production. Recombinant virus products induced antibody formation in dogs to human influenza-A virus, herpes simplex virus, and human hepatitis-B virus antigens. It was concluded that vaccinia virus would be safe and suitable as an eukaryotic expression vector in dogs. PMID- 2977719 TI - [Valvuloplasty in pulmonary and aortic stenosis and angioplasty in aortic coarctation]. PMID- 2977720 TI - The epidermal G1-chalone: an endogenous tissue-specific inhibitor of epidermal cell proliferation. AB - An apparently macromolecular factor is isolated from aqueous skin extracts which inhibits DNA synthesis in vivo and in vitro with high efficacy (ID50 in vivo 0.2 pmol/g, in vitro 0.2 pM) and in a highly specific manner showing a point of attack in the late G1-phase of the cell cycle (epidermal G1-chalone). Preliminary characterization indicates an unusual highly amphipathic structure consisting of amino acids and carbohydrate. Despite its apparent molecular weight of approximately 10 kD the chalone is stable against denaturing agents and most enzymes, including proteases. An inverse correlation between chalone responsiveness of mouse epidermis in vivo and the development of hyperplasia due to injury indicates an important role of the factor in the regulation of tissue homeostasis. According to its physicochemical and biological properties the epidermal G1-chalone appears not to be related to other endogenous inhibitors of epidermal cell proliferation such as the pentapeptide pyroGlu-Glu-Asp-Ser-GlyOH and transforming growth factor beta (TGF beta). PMID- 2977721 TI - Assignment of complement components C4 binding protein (C4BP) and factor H (FH) to human chromosome 1q, using cDNA probes. AB - Using cDNA probes for Factor H (FH) and C4 binding protein (C4BP) on a panel of somatic cell hybrids, we show that both of these genes map to the long arm of chromosome 1. PMID- 2977722 TI - [Platelet activation during exacerbation of bronchial asthma--thrombin induced ATP release from washed platelets and plasma beta-TG, PF4, beta-TG/PF4 ratio]. PMID- 2977723 TI - [Coronary transluminal angioplasty: current indications and results]. PMID- 2977724 TI - Decrease in arterial blood pressure in spontaneously hypertensive rats by the TXA2 receptor antagonist BM 13177 and by acetylsalicylic acid. AB - The effects of the selective TXA2 receptor antagonist BM 13177 and of the cyclooxygenase inhibitor acetylsalicylic acid on blood pressure and heart rate were investigated in anaesthetized spontaneously hypertensive rats (SHR). After intravenous injection of BM 13177 (60 mg/kg) there was a strong short-lasting decrease in arterial blood pressure (BP) with a maximum reduction of 21.6% in the third minute after injection. The heart rate was only slightly decreased. The formation of TXB2 and 6-keto-PGF1 alpha in serum, aorta thoracica, renal medulla, and renal cortex, which was determined by radioimmunoassay, was not altered by BM 13177. Pretreatment with acetylsalicylic acid (100 mg/kg) attenuated the BM 13177 induced reduction of blood pressure and heart rate in male, but not in female SHR. An intravenous application of acetylsalicylic acid (ASA) (100 mg/kg) resulted in a decrease of blood pressure and heart rate only in male rats. The extent of these effects was similar to that induced by BM 13177. A possible explanation for the different reactions of male and female animals might be sex differences in the sensitivity of cyclooxygenase in the aorta thoracica to ASA and/or in the balance of prohypertensive and antihypertensive prostanoids. TXA2 and other prostanoids seems to participate in the short-time regulation of BP in SHR. PMID- 2977725 TI - Smooth muscle effects of hydroxylated docosahexaenoates produced from human platelet. AB - Washed platelets (10(8)cells/ml) are capable of metabolizing docosahexaenoic acid (22:6w3, DHE) to 12-lipoxygenase derivatives. The two major metabolites of the DHE thus formed were collected and derivatized for analysis by GC/MS/EI. The structures assigned were 14(S) and 11(S) hydroxy-docosahexenoate (HDHE). 12 lipoxygenase inhibitors such as ETYA inhibited the production of HDHE. The metabolites formed are biologically active as they are capable of inducing a weak contraction in airway but not vascular smooth muscle preparations; a thromboxane agonist (U46619) was 10 to 20-fold more efficacious than HDHE in the guinea pig lung parenchymal strip. HDHE may act in part through stimulation of leukotriene production as increased peptidyl-leukotriene levels were associated with the HDHE induced contraction in this preparation and a lipoxygenase inhibitor (NDGA) was capable of a partial blockade of this response. In addition, HDHE antagonizes the contractile effects of the thromboxane-agonist, U46619, especially in vascular smooth muscle. Stimulation of the sulfidopeptido leukotrienes and thromboxane antagonism may therefore be important aspects of the biological function of HDHE. PMID- 2977727 TI - Moyamoya and other causes of stroke in patients with Down syndrome. AB - Twenty-seven new cases of stroke in Down syndrome are reported, two probands, 13 patients from review of institutional records, and 12 patients from a survey of child neurologists. Forty-one patients from previous reports are summarized. Most stroke episodes were related to congenital heart disease and infections. Seven cases were associated with angiographic abnormalities; there were three cases of unknown etiology and four cases of moyamoya. The understanding of moyamoya and other causes of stroke may be advanced by further study of stroke in Down syndrome patients. PMID- 2977726 TI - [Peroxidative vulnerability of synaptosomal high affinity Ca++-ATPase and pharmacologic effects]. AB - The high affinity Ca++-ATPase participates essentially in the regulation of intrasynaptosomal calcium homeostasis. Related to posthypoxically restricted transmitter release, we examined the influence of newly-generated free radicals (ascorbic acid-ferric salt mixture) or sodium dodecyl sulfate in vitro and of a mild hypobaric hypoxia in vivo on the activity of synaptosomal high affinity Ca++ ATPase. Moreover we tested the effectiveness of piracetam, meclofenoxate hydrochloride, pyritinol and verapamil on the changed enzyme activity subsequent to a hypoxic exposure. The activity of synaptosomal high affinity Ca++-ATPase (1.04 +/- 0.03 mumol Pi/mg.h) is reduced by not more than 40% depending on the concentration of the ascorbic acid-ferric salt mixture used but is nearly totally inhibited by sodium dodecyl sulfate (0.2 mg/ml). Hypobaric hypoxia (18 h, 8.7 kPa) decreases the enzyme activity to 0.79 +/- 0.03 mumol Pi/mg.h. Piracetam, meclofenoxate hydrochloride and pyritinol are protectively effective on the decrease of enzyme activity induced by hypoxia. The results emphasize the importance of intact protein-phospholipid interactions for the enzyme activity and support relations between synaptosomal high affinity Ca++-ATPase and transmitter release. PMID- 2977728 TI - Spontaneous onset of ciliary block glaucoma in acute hydrops in Down's syndrome. AB - A flat anterior chamber and raised intraocular pressure are unusual complications of acute hydrops. We report a case in which a patient with Down's Syndrome developed a flat anterior chamber, together with raised intraocular pressure, several weeks after the onset of acute hydrops. PMID- 2977729 TI - [The role of structural changes in T4 bacteriophage tail proteins]. AB - Conformational changes in bacteriophage tail proteins after heating and ionic strength alteration leading to dissociation of tail sheath have been studied using protein fluorescence, differential scanning microcalorimetry and electron microscopy methods. Autonomous structural changes in tube-baseplate proteins have been revealed. They take place under the same conditions as those which release the bonds holding the sheath protein subunits to those of the tube in isolated sheathed tails. The conformational changes in the tube-baseplates are reversible similarly to the process of assembly and disassembly of the extended sheath. Morphological changes in the tube have been found at the temperature above the transition registered by protein fluorescence but not by calorimetry. This suggests that revealed spectral alterations reflect changes in quaternary structure of tail tube in particular. PMID- 2977730 TI - Calcium uptake and inositol trisphosphate-induced calcium release from parathyroid gland membranes. AB - The manner in which changes in Ca2+ are translated by the parathyroid gland into a signal for secretion of its major secretory proteins, parathormone and secretory protein-I, is not understood. In order to gain insight into this mechanism, Ca2+ transport was evaluated in porcine parathyroid gland membranes prepared by homogenization and differential centrifugation. The membranes were considered to be intact and appropriately 'sided' since they contained secretory protein-I and exhibited ATP-dependent uptake of [45Ca2+]. About 1 mol of ATP was hydrolyzed per mol of Ca2+ transported, signifying that the ATPase activity was relevant to the Ca2+ transport system of the membranes. Inositol trisphosphate, which is believed to be a universal signal for generating a change in cytosolic Ca2+ by discharging Ca2+ from internal membranes, caused the parathyroid membranes to release accumulated Ca2+. The membranes were able to partially reaccumulate the discharged Ca2+. The data suggest that intracellular Ca2+ metabolism in the parathyroid cell is similar to that of other secretory cells. If a change in intracellular concentration of this ion is involved in secretion, it may occur locally within the cytosol. PMID- 2977731 TI - Neutrophil function in megaloblastic anemia. AB - Chemotaxis, nitroblue tetrazolium (NBT) reduction and the presence of Fc gamma receptors on surface membranes were measured in the peripheral blood neutrophils of 20 patients with megaloblastic anemia (15 with folate deficiency and 5 with pernicious anemia). The percent of NBT- or Fc gamma-positive neutrophils was the same for patients and normal controls. In contrast, chemotaxis was decreased approximately 20% (P less than 0.05) in megaloblastic anemia and when anemia was corrected in 3 cases, chemotaxis increased to levels within the normal range. In contrast to what one would expect on the basis of these laboratory findings, there are no reports of increased susceptibility to infections in these patients. PMID- 2977732 TI - [Treatment of precocious puberty with an LH-RH agonist (D-TRP6-LH-RH delayed release microcapsules)]. PMID- 2977733 TI - [Intraluminal coronary angioplasty]. PMID- 2977734 TI - Two-step treatment approach for periodontitis in AIDS patients. PMID- 2977735 TI - Managing the oral manifestations of cancer therapy, Part I: Head-and-neck radiation therapy. PMID- 2977736 TI - Managing the oral manifestations of cancer therapy, Part II: Chemotherapy. PMID- 2977737 TI - Dental management of the diabetic patient. PMID- 2977738 TI - Nalbuphine hydrochloride: an alternative parenteral analgesic for patients with a compromised respiratory system. PMID- 2977740 TI - Anatomical deformations due to footwear in Zimbabwe. PMID- 2977739 TI - ARC patient sues dentist for treatment refusal. PMID- 2977741 TI - An inside look at pediatric dentistry. Interview by Tope Maxson. PMID- 2977742 TI - [Diagnosis of neck, arm, lumbar and leg pain by electromyography of the cervical and lumbar rotator brevis muscles]. PMID- 2977743 TI - [Clinical significance of cardionatrin in the diagnosis of deficiency of heart lung qi]. PMID- 2977744 TI - T lymphocytes in Graves' disease. PMID- 2977745 TI - Clinical evaluation of three sanguinarine delivery systems for use in a psychiatric hospital preventive dentistry program. PMID- 2977746 TI - Influence of dietary lipids on microsomal membranes from chick breast muscle. AB - 1. The effect of different dietary fat intake on the lipid composition and fluidity of microsomal membranes as well as in the enzymatic activity of the Ca2+ ATPase from chick breast muscle was investigated. 2. When a standard diet was supplemented with 10% sunflower seed oil, an increase in the relative amounts of unsaturated fatty acids and membrane fluidity and a decrease in the cholesterol content was observed. 3. The presence of 6% cholesterol in the diet does not modify the fatty acid composition and the fluidity of the membrane but increased, in a low extension, the cholesterol content. 4. The provision of the sunflower seed oil-rich diet supplemented with cholesterol just 48 hr before death promoted an increase in the relative amounts of unsaturated fatty acids and cholesterol content whereas the membrane fluidity decreased in a significant extent. 5. Despite that dietary lipids gave rise in some cases to changes in lipid composition and in the physical state of the microsomal membrane, neither the Ca2+ uptake capacity nor the ATPase activity were significantly affected. PMID- 2977747 TI - Inhibitory effect of squid (Todarodes pacificus) paramyosin on actomyosin ATPase and on superprecipitation. AB - 1. Paramyosin from squid mantle muscle inhibited the Mg-ATPase and the superprecipitation activities of actomyosin. 2. The inhibition was detected only when paramyosin forms a cofilament with myosin. 3. ATP-induced changes in the morphology of the cofilament of myosin and paramyosin are involved in the inhibition by paramyosin. PMID- 2977749 TI - No evidence for linkage between lipoprotein(a) (LPA) and esterase D (ESD). PMID- 2977748 TI - Thick filaments of fish myosin and its actin-activated Mg-ATPase activity. AB - 1. Tilapia (Tilapia nilotica) myosin forms short, mini-filaments, and are easily disassembled upon addition of ATP showing no saturated activation in its actin activated Mg-ATPase activity. 2. The presence of 5-10 mM MgCl2 allows tilapia myosin to form native thick-filaments and are resistant to ATP. 3. The rod portion of Tilapia myosin molecule is responsible for its characteristic filament forming ability. 4. The similar filament forming ability as Tilapia myosin was suggested for other fish myosins. PMID- 2977750 TI - [The stomatological treatment of diabetic children]. PMID- 2977751 TI - [Mechanism of cell damage by hematoporphyrin derivative (HPD) plus light. II. Effect of HPD plus light on respiration and oxidative phosphorylation in hepatoma cells and normal liver mitochondria]. AB - The effects of HPD plus light on energy metabolism and membrane structure of hepatoma cell and normal liver mitochondria were investigated. HPD was bound to mitochondria either in vitro or in vivo. Mitochondria bound HPD plus light increased MDA level, decreased respiratory intensity and respiratory control, dissociated phosphorylation from oxidation and enhanced mitochondria swelling on age. These effects were related to HPD concentration and exposure time but it was unchanged in the control. These results showed that the loss of function of oxidation phosphorylation was due to the mitochondrial membrane system damage under the action of singlet oxygen produced by HPD bound mitochondria following light activation. PMID- 2977752 TI - [An analysis of iatrogenic coronary artery dissection in 5 cases]. PMID- 2977753 TI - [Plasma beta-thromboglobulin and platelet factor 4 in essential hypertension]. PMID- 2977754 TI - [Normal values of total 12-lead QRS voltage and its significance in diagnosing left ventricular hypertrophy]. PMID- 2977755 TI - Endodontic treatment and immunopathology of periapical granuloma in an AIDS patient. PMID- 2977756 TI - [Creatine kinase and aspartate and alanine transaminase activity in the myocardium and blood serum of albino rats in experimental streptozotocin diabetes mellitus. Effect of (-)-epicatechin]. PMID- 2977757 TI - Isolation and structure determination of the main related substances of teicoplanin, a glycopeptide antibiotic. AB - Teicoplanin is a complex formed by five closely related glycopeptides and by a small amount of a hydrolysis product. Minor quantities of related substances are also present. Two of them (named RS-1 and RS-2) were isolated and purified starting from the tailing fractions of a teicoplanin batch. Preparative reversed phase liquid chromatography on large low-pressure and medium high-pressure scales, concentration, desalting, and freeze-drying steps were applied. 300 mg of RS-1 and 900 mg of RS-2 were obtained in a purity grade (about 90%) sufficient for structural investigation. Starting from considerations on the HPLC retentivity and on biosynthesis, the structures were assigned on the basis of 1H N.M.R. spectra and homonuclear CO-SY 2D experiments, FAB-MS spectrometry, and GC MS of the esters of the fatty acids obtained by hydrolysis. RS-1 and RS-2 are teicoplanins having 10-methyl-undecanoic acid and n-dodecanoic acid, respectively as fatty acid chains. No major difference in the in vitro activity of these teicoplanins emerged in comparison with teicoplanin complex. PMID- 2977758 TI - [Contents of macroergic phosphates, ATPase and creatine kinase activity of tissues of the artery and vein walls in rabbits]. PMID- 2977759 TI - Immunoregulatory granulocyte factor (GF). II. In vitro features of antibodies against GF+. AB - Activated polymorphonuclear leucocytes (PMNL) secrete a specific factor possessing numerous immunoregulatory properties. The rabbit IgG antibodies against this human granulocyte factor GF (anti-GF-IgG) have significantly changed the regulatory features of GF and the same antibodies alone have also exerted immunoregulatory potency when tested in vitro. Inhibition of mixed lymphocyte reaction (MLR) proliferation after GF treatment was abolished in the presence of anti-GF-IgG. GF displays a potentiating effect on interleukin-1 (IL-1) production by human monocytes in vitro. Anti-GF-IgG did not change the potentiating capacity of GF and these specific antibodies alone also enhanced IL-1 production to the same extent as GF. In peripheral blood mononuclear cell (PBMC) cytotoxicity test, the cytotoxic activity of effector cells was markedly reduced in the presence of GF. Anti-GF-IgG partially abrogated the observed suppression, although their own features indicate a strong inhibitory effect on PBMC cytotoxicity. These contradictory results are difficult to explain but they may enable us to understand the pleiotropic immunoregulatory mechanism of both GF and anti-GF-IgG. The results of the present and recently published studies support the idea that both GF and anti-GF-IgG modify T lymphocyte function at the same receptor level. PMID- 2977760 TI - Expression of major histocompatibility complex class II antigen on target cells in autoimmune endocrine diseases. PMID- 2977761 TI - Chairside dental assisting techniques for the disabled student. PMID- 2977762 TI - Hematoporphyrin ethers--II. Improvements in method, synthesis of the dihexyl, dicyclohexanyl and diphenyl ethers and their preliminary biological evaluation. AB - 1. Hematoporphyrin 2.4-dihexyl, -dicyclohexanyl and -diphenyl ethers have been synthesized. 2. Their chemical and physical properties are recorded. The possibility of isomerism is discussed. 3. Preliminary tests have indicated that they possess high photosensitizing efficiency on human cancer cells. PMID- 2977763 TI - Preparation and characterization of biotinylated probes for the beta-interferon receptor. AB - 1. Recently we described the isolation of the beta-interferon receptor [Zhang et al. (1986) J. biol. Chem. 261, 8017-8021]. A highly purified product was obtained but in low quantities. 2. The use of biotinylated beta-interferon as a ligand represents an alternate approach to receptor isolation. 3. We have prepared and characterized the derivatives N-(biotinyl)- and N-(biotinyl-epsilon-aminocaproyl) recombinant human [Ser17]-interferon beta (B- and BC-recHuIFN beta). 4. Biotin incorporation does not result in any loss of antiviral activity, demonstrating the recognition of the derivative by the cell receptor. 5. The biotinylated recHuIFN beta binds specifically and reversibly to succinoylavidin or guanidine thiocyanate-stripped succinoylavidin linked to a Sepharose matrix. 6. Comparison of the competition curves obtained with [14C]biotin and [3H]biotinyl recHuIFN, in the presence of increasing concentrations of biotin suggests that the IFN moiety of the derivative has little effect on the affinity of biotin for avidin. 7. Biotinylated recHuIFN beta derivatives represent useful probes for the beta-IFN receptor. PMID- 2977764 TI - Fructose-2,6-bisphosphate in rat mesenteric lymph nodes. AB - 1. The fructose-2,6-bisphosphate (Fru-2,6-P2) content of mesenteric lymph nodes was measured in rats. 2. The effects of Fru-2,6-P2 on the activity of 6 phosphofructo-1-kinase (PFK-1) from rat mesenteric lymph nodes were also studied. 3. The affinity of the enzyme for fructose-6-phosphate was increased by Fru-2,6 P2 whereas the inhibition of the enzyme with high concentrations of ATP was released by Fru-2,6-P2. 4. The activity of lymphocyte PFK-1 was highly stimulated in a simultaneous presence of low concentrations of AMP and Fru-2,6-P2. 5. These results show that rat lymphocyte PFK-1 is highly regulated with Fru-2,6-P2 which means that glycolysis in rat lymphocytes is controlled by Fru-2,6-P2. PMID- 2977765 TI - A simple purification method for a glucosyltransferase complex from Streptococcus mutans OMZ 176 with a high yield. AB - 1. Glucosyltransferase was purified from Streptococcus mutans OMZ 176 by ammonium sulfate fractionation and gel filtration. 2. The overall purification was about 15,000-fold with a yield of 53%. 3. The purified enzyme appeared to be a multienzyme complex consisting of three different components with molecular weights of 158,000, 153,000 and 149,000, respectively. PMID- 2977767 TI - In vitro caries induction at the tooth-amalgam interface. PMID- 2977766 TI - Silver release from metal-reinforced glass ionomers. PMID- 2977768 TI - [Aortic valvuloplasty associated with coronary angioplasty. Description of a case]. AB - The authors report the case of a 57 year-old man with severe calcific aortic stenosis and single vessel coronary artery disease, symptomatic of angina pectoris, who refused surgical therapy and was treated with combined percutaneous aortic valvuloplasty and coronary angioplasty as a single procedure. Valvuloplasty was performed initially, and the calculated aortic valve area increased from 0.75 to 1.27 cm2. Subsequently, a 75% proximal right coronary artery stenosis was dilated to 20%. Two months later, at follow-up the patient was asymptomatic and the Doppler-determined aortic valve area was 1.01 cm2. PMID- 2977769 TI - [Isolated enlargement of the right atrium. Description of a successfully operated case]. AB - We report a case of isolated right atrial enlargement. The patient was an 11 year old boy who had life-threatening supraventricular arrhythmias. Diagnosis was basically performed by Echo-2D Doppler. The surgical treatment performed on cardiopulmonary by-pass, was followed by immediate conversion to sinus rhythm. Histological examination showed chronic inflammatory aspects. Postoperative Echo 2D (2 years later) showed a moderate right atrial enlargement. PMID- 2977770 TI - DNA methylation and control of genome organization in Neurospora crassa. PMID- 2977771 TI - Methylation-dependent gene expression in phage Mu. PMID- 2977772 TI - [The role of occupational and various social factors in the development of diseases among workers in the chemical fiber industry]. PMID- 2977773 TI - Acquired immune deficiency syndrome. What we know--what questions remain. PMID- 2977774 TI - Responsibility of the dental team: prevention and patient education. PMID- 2977775 TI - Examination, detection, diagnosis and referral. PMID- 2977776 TI - Dental management of the head and neck cancer patient. PMID- 2977777 TI - [Dermatitis caused by bulbar analeptics: mepixanox and theophylline]. PMID- 2977778 TI - State of electrolytes, osmotic balance and the activity of ATPase in the lenses of selenite--induced cataracts. AB - Selenite-cataracts incorporated many morphological characteristics observed in human senile catracts. Progressive elevation of sodium, marked loss of potassium, several fold increment of calcium; considerable loss of magnesium levels, a dose response reduction of total-ATPase activity and significant hydration are the important features observed in the lens during the progressive treatment of selenite. The sodium-potassium imbalance is found to be a secondary effect during the development of cataract and is suggested to bring about by an abnormal accumulation of calcium ions and inactivation of transport enzyme. The calcium activated proteases could be the promoting factor for the proteolysis and insolubilization of lens proteins in the inducement of selenite cataract. The impact of selenite on the SH containing ATPase anzymes could be the cause of impairment in energy metabolism, derangement of electrolytes and osmotic imbalance which, in turn, accelerate the cortical involvement of lens opacities. PMID- 2977779 TI - Alterations in T-helper and T-suppressor lymphocyte populations after multiple injuries. AB - Fifteen patients with multiple injuries and an Injury Severity Score (ISS) ranging from 2 to 57 (median 25) were studied for variations in lymphocyte populations on the day of injury and the three following days. Nine of the patients had an ISS above 16. In all patients the total number of lymphocytes fell during the first 24 hours after the injury (P less than 0.01), mainly due to a reduction in the number of circulating T-lymphocytes from a median of 1.8 to 0.6 x 10(9) cells/l (P less than 0.01). No reduction in the T-helper/T-suppressor cell ratio could be demonstrated for the group as a whole, but in the 9 patients with an Injury Severity Score (ISS) of more than 16 a significant reduction in ratio from median 1.5 to 0.8 was found. Due to clinical observations patients with an ISS of more than 16 are considered severely injured and at risk of developing complications such as infection and septicaemia. The fall in T helper/T-suppressor ratio indicates impaired immunity in these patients. PMID- 2977780 TI - Disembowelment--a retrospective study of patients suffering evisceration following penetrating abdominal injury. AB - Details are presented of 104 patients admitted over a 1-year period with evisceration following penetrating abdominal wounds. Stabbing was the most common cause of injury. Small bowel was the most commonly eviscerated organ, as well as the organ most frequently injured. Exploratory laparotomy was performed on all the patients who survived to reach the operating theatre. The negative laparotomy rate was 12 per cent, and a further 3 per cent had such mild visceral injuries that they did not require operative repair. For comparison a group of 111 patients admitted over the same period, but suffering only omental protrusion, was also examined. The rate of visceral injury in this group was slightly lower. Wound complications, including breakdown, were more commonly seen in the eviscerated patients, but the mortality was 8 per cent in both groups. PMID- 2977781 TI - Report addresses ethical ramifications of refusing to treat patients with AIDS or patients who are HIV seropositive. PMID- 2977782 TI - Tetraethylammonium-induced contraction of rabbit coronary artery. AB - The vasoactive effect of tetraethylammonium (TEA), a well-known K channel blocker, was tested on helical strips excised from the large epicardial coronary arteries of rabbit hearts. TEA (10 mM) induced transient tetanic contraction of greater amplitude as a result of summation of twitch responses. Occasionally, spontaneous periodic contractions occurred during prolonged exposure to 10 mM TEA. This TEA-induced contraction was abolished in Ca-free solution and suppressed by Ca-entry blockers: nitroglycerin, nicorandil, and isoproterenol, but not by phentolamine or atropine. In strips in which TEA did not induce remarkable contraction, subsequent addition of a subthreshold concentration of ergonovine, serotonin, acetylcholine, ouabain, K-rich solution, or alkalinization of the solution provoked remarkable contraction. These results are consistent with previous reports that TEA induced tetanic contraction as a result of summation of twitch responses due to spontaneous discharge of Ca-spikes in some arterial and tracheal smooth muscle. The results also support the idea that TEA induced contraction of the rabbit coronary artery is mediated by the same mechanism, i.e., spontaneous Ca-spike discharge. PMID- 2977784 TI - Atrial natriuretic factor. PMID- 2977785 TI - Treatment of severe psoriasis with low dose cyclosporin A and the effect on the helper-suppressor T cell ratio in peripheral blood. PMID- 2977783 TI - Impaired myocardial perfusion in patients with hypertrophic cardiomyopathy: assessment with digital subtraction coronary arteriography. AB - To study the clinical significance of abnormal myocardial perfusion in patients with hypertrophic cardiomyopathy (HCM), we performed a computerized washout analysis of digital subtraction coronary arteriograms in 28 patients with HCM and 16 control subjects. The contrast disappearance half-life (T1/2) was calculated from a time-density curve generated in the four sectors of the myocardium perfused by the left anterior descending coronary artery and the mean T1/2 was calculated by averaging T1/2 values for these four sectors. Patients with HCM demonstrated longer T1/2 in the ventricular septal region than control subjects. Thirteen (46%) of the patients with HCM presented abnormally longer mean T1/2 values, suggesting impaired myocardial perfusion. Family histories of HCM were more frequent in patients with abnormal mean T1/2 values (92% vs 47%; p less than 0.05). On the exercise stress test, patients with abnormal T1/2 values presented significantly lower exercise tolerance with more frequent exercise-induced ST segment depression (62% vs 13%; p less than 0.05). However, there were no significant differences between the two groups with regard to ventricular wall thickness, left ventricular end-diastolic pressure, or the severity of systolic narrowing of the coronary arteries. These findings suggest that 13 (46%) of the patients with HCM have impaired myocardial perfusion, which may be a manifestation of intramural coronary artery disease in addition to left ventricular hypertrophy, elevated left ventricular end-diastolic pressure, or systolic narrowing of the coronary arteries. Additionally, significant association of the prolonged T1/2 with a familial occurrence of HCM and depressed exercise tolerance with ST segment depression imply that impaired myocardial perfusion could be an important inherent pathophysiological state leading to myocardial ischemia during exercise. PMID- 2977786 TI - The monoclonal antibody-defined CAR-3 antigen is a serological marker associated with pancreatic carcinoma. AB - The monoclonal antibody-defined CAR-3 antigen is a new carcinoma associated marker which is expressed on a mucin-like molecule. Serum concentrations of CAR-3 were assayed in 181 patients with carcinomas of different organs, 20 patients with non-carcinomatous malignancies, 123 patients with inflammatory diseases and 150 healthy controls. Serum levels of CAR-3 were significantly increased in 51% of the patients with pancreatic carcinomas, in 60% of patients with biliary tract carcinomas and in about 15% of the patients with carcinomas of the digestive apparatus. Sera from patients with breast carcinomas were negative, as well as sera from patients with melanomas or sarcomas. CAR-3 values in samples from patients with chronic pancreatitis were constantly negative, as were samples from healthy donors. Significant concentrations of CAR-3 were detected in 20% of the sera from patients with acute pancreatitis and in 15% of the sera from patients with cirrhosis. Because of its high specificity for pancreatic carcinomas compared to chronic pancreatitis, CAR-3 seems a promising marker for distinguishing between neoplastic and chronic inflammatory diseases of the pancreas, whose differential diagnosis is difficult. PMID- 2977787 TI - [Biochemistry of dynein, the ATPase from cilia or flagella]. PMID- 2977788 TI - Endodontic therapy for mental nerve neuropathy in systemic cancer patients. PMID- 2977789 TI - Inhibition of Streptococcus mutans glucosyltransferase by M-GTFI, a new inhibitor. AB - Two hundred strains of soil microorganisms were screened for the production of inhibitors of the glucosyltransferase activity of Streptococcus mutans strain, K1 R. The strain producing the greatest amount of inhibitor was one recently isolated in our laboratory. It has now been identified as a strain of Micromonospora narashinoensis on the basis of morphological and physiological studies. The inhibitor, M-GTFI, affects the glucosyltransferase that produces the water-insoluble glucan rather than that which produces the water-soluble glucan. Fuchsin-sulphite staining of the inhibitor after its purification by polyacrylamide gel electrophoresis indicates that it is probably an acidic substance. It had Mr 5700 as was determined by gel filtration. From an examination of the effects of this inhibitor on representative strains of S. mutans other than K1-R, there is a suggestion of a similar selectivity for the water-insoluble glucan-forming activity in other strains. PMID- 2977791 TI - [Evaluation of coronary blood flow using digital subtraction technique and cine coronary angiography: a preliminary report]. AB - To evaluate coronary circulation in ischemic heart disease, digital image processing with cine coronary angiography was performed. Using time-density curves obtained from individual pixels, images showing the distribution of contrast density and transit time were obtained. To record angiograms, contrast medium was injected into the coronary artery in a steady manner during right atrial pacing. Frames in the end-diastolic phase immediately before the P wave were selected, and digitized into a 512 x 512 x 8 bit matrix using a system composed of a video camera, an analog-to-digital converter, and a computer. These digitized images were then stored in a disk memory. A mask image was prepared before the injection of contrast medium. Subtraction was performed using the mask image and a series of images following contrast injection. The subtracted images were of sufficient quality to permit clear observation of the individual coronary arterial branches. Time-density curves were then determined from these subtracted images. From these curves, time from the onset of contrast injection to its peak density (Tp), time from the peak density to the half peak density (T 1/2) and the attenuation factor of the curves (tau) were derived. Their distributions were expressed as color images. Examples of a normal control and a case of inferior infarction were demonstrated. Blood flow function images with good spatial resolution were thus obtained. This method is useful for evaluating coronary blood flow. PMID- 2977790 TI - Immune properties of glucosyltransferases from S. sobrinus. AB - GTF activity was separated into water-insoluble (GTF-I) and water-soluble (GTF-S) polyglucan-synthesizing enzymes. Each preparation demonstrated a single band on 6% SDS PAGE. Only water-insoluble or water-soluble polyglucan was synthesized by the respective enzyme preparation. Rats were immunized, on Days 1 and 14, with either GTF-I or GTF-S in adjuvant. Animals were bled 13, 35 and 54 days after the initial immunization. Individual antisera were tested against either the GTF-I or the GTF-S for inhibition of radioactive glucose incorporation into glucan, and in gel diffusion, and by Western transfer analyses. The respective antisera reacted with the homologous, but not the heterologous enzyme in gel diffusion and Western transfer. GTF-I activity was not inhibited by antibody to GTF-S, but antibody to GTF-I inhibited GTF-I by 68%. GTF-S was inhibited by more than 60% by each of 3 anti-GTF-S sera. Only one anti-GTF-I serum inhibited GTF-S at as much as a modest 30% level. These data support the antigenic and functional distinctiveness of the GTF enzymes of S. sobrinus 6715. PMID- 2977792 TI - [Tissue characteristics in left ventricular hypertrophy using magnetic resonance imaging]. AB - For 15 normotensive patients with asymmetric septal hypertrophy (ASH), 10 hypertensive patients with concentric hypertrophy (CH), and five normal subjects (N), we examined changes in myocardial T1 and T2 values related to the cardiac cycle. The usefulness of those values in differentiating diseases with left ventricular hypertrophy was evaluated. Left ventricular (LV) short-axis spin echo images and inversion recovery images were obtained at endsystolic and diastolic cardiac phases, and T1 and T2 images were calculated. The regional wall thickness (WT) and T1 and T2 values were measured in the anterior septum, anterior wall, lateral wall, posterior wall and posterior septum. Myocardial T1 and T2 values were significantly decreased in systole (T1: 185.6 +/- 37.9 msec, T2: 24.4 +/- 6.3 msec, mean +/- SD) compared to those in diastole (T1: 249.2 +/- 56.7 msec, T2: 31.7 +/- 9.4 msec). In both the ASH and CH groups, significant correlations were observed between diastolic T1 values and WT (ASH: r = 0.80, p less than 0.01, CH: r = 0.45, p less than 0.01), and between diastolic T2 values and WT (ASH: r = 0.58, p less than 0.01, CH: r = 0.60, p less than 0.01). In the regions where diastolic WT were more than 17 mm, T1 values in the ASH group (343.4 +/- 40.5 msec) were significantly higher than those of the CH group (247.3 +/- 21.4 msec), although the mean wall thickness values were similar in both groups. The T1/WT and T2/WT were significantly lower in the CH group than those in the ASH and N groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977793 TI - [Diastolic dysfunction and its relation to myocardial fibrosis in essential hypertension]. AB - This study elucidated diastolic left ventricular dysfunction and whether myocardial interstitial fibrosis correlates with diastolic dysfunction in mild to moderate systemic hypertension (HT). Six normotensive subjects, 18 hypertensive patients without left ventricular hypertrophy (LVH) and 10 hypertensive patients with significant LVH were evaluated. M-mode echocardiography was used to determine fractional shortening (FS), isovolumic relaxation time (IRT), and left ventricular filling volume during rapid and slow filling periods and the atrial contraction period (RFV, SFV, ACV). The quotients of the left ventricular filling volume and the end-diastolic volume and stroke volume were also calculated. Simultaneous biventriculography was used to determine the end-diastolic thickness of the interventricular septum and posterior wall. Right ventricular endomyocardial biopsies were performed to calculate the percentages of fibrosis. The FS was normal in all groups. The percentages of fibrosis in the two HT groups were significantly greater than those in the normals. The IRT of the HT groups was significantly greater, and the RFV, RFV/EDV and RFV/SV were significantly less than those of the normals. Multiple regression analysis showed that the wall thickness and the percentages of fibrosis correlated significantly with IRT, RFV, RFV/EDV and RFV/SV. The standard coefficients of correlation of wall thickness and the percentages of fibrosis were 0.333 and 0.239 in respect to IRT, and 0.304 and -0.473 in respect to RFV. There were significant correlations between the percentages of fibrosis and RFV (r = -0.675), RFV/EDV (r = -0.664) and RFV/SV (r = -0.602) in the normals and in cases of HT without LVH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977794 TI - [Morphological problems in cardiac hypertrophy]. AB - Ventricular hypertrophy is categorized as (1) volume hypertrophy (eccentric hypertrophy or hypertrophy with dilatation), and (2) pressure hypertrophy (concentric hypertrophy or hypertrophy without dilatation). Hypertrophy per se is considered as a reaction to hemodynamic overload, both in occasions of excessive volume and of elevated pressure except in (idiopathic) cardiomyopathies. In patients with volume hypertrophy, some alteration of myocardial architecture is inevitable if the volume load exceeds a critical degree. Such alteration is characterized by deterioration of the median circular muscular layer of the ventricle resulting from over-stretching of the horizontally-coursing myocyte bundles. Hypertrophy of the inner oblique muscular layer of the left ventricle in cases with mitral regurgitation, and of the outer oblique muscular layer in cases with aortic regurgitation develops according to the changing configuration of the left ventricle; globular in the former, and elongated in the latter. In patients with pressure hypertrophy, there is some increase of the myocyte mass, with a disarrangement at the anterior triangle of the interventricular septum. The latter is physiologically situated at the anterior margin of the septum between the outer and median layers of both the ventricles. This seems to be a prelude to thickening of the septum and tends to hypertrophy of the free wall. A reduction in the deranged myocyte mass due to fibrosis or adiposis corresponds to arrest of the active hypertrophic process, and it may be the beginning of decompensation of cardiac function. Peculiar modes of dilatation and hypertrophy in cardiomyopathies have common denominators with those of known etiologies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977795 TI - [Coronary artery blood flow velocity non-invasively measured using a vessel tracking pulsed Doppler system]. AB - A newly-developed noninvasive method was used to measure left coronary blood flow during phantom experiments. Two techniques were used in which: (1) the sample position can always be set in a fluctuating vessel using a wall echo-tracking method with a phase-locked-loop, and (2) the Doppler reference signal was generated separately synchronous with the wall echo signal. These techniques were combined, using a commercially available pulsed Doppler apparatus (SSH-40B: Toshiba). Basic experiments were performed using a blood vessel phantom to verify the validity of these systems. Blood flow velocity in the fluctuating tube could be measured clearly using a vessel-tracking method. The blood flow velocity of the left anterior descending artery was measured in three normal subjects and in seven patients from the third intercostal space along the left sternal border. The velocity pattern was characterized by a crescendo-decrescendo shape in diastole. The peak velocity which appeared in diastole ranged from 19 to 69 cm/sec, with no difference by disease entity. However, in all cases, the blood flow velocity signals were marred by extraneous signals, making it impossible to measure blood flow velocity during systole. Further improvement of the system is mandatory in order to use this flowmeter clinically. PMID- 2977796 TI - [Analysis of myocardial texture in two-dimensional echocardiographic images]. AB - To evaluate tissue changes, we studied myocardial texture using two-dimensional echocardiographic images. We investigated 19 normal subjects, 28 patients with left ventricular hypertrophy, and 12 patients with old anteroseptal myocardial infarction of longer than one year duration. Using 2.5, 3.5, and 5.0 MHz transducers, two-dimensional echocardiograms in the parasternal long-axis view were obtained, and the textures of the interventricular septal images were classified in three types; type I, with a nearly uniformly speckled or echolucent appearances; type II, with multiple, discrete, small (2 to 4 mm) highly refractile echoes; type III, with larger highly refractile echoes (greater than 4 mm) appearing as a cluster of broad patches or band-like echoes. Normal subjects belonged to the type I texture, while many with left ventricular hypertrophy belonged to the type II category. Type III was often observed in patients with old anteroseptal infarction. Using a transducer of higher frequency, there tended to be a shift from type II to I or type III to II. In phantom experiments using carbolandam granules instead of the myocardium, the echocardiographic texture became rough when the phantom was farther from the transducer or the transducer was of low frequency. We suggest that the texture in two-dimensional images may reflect myocardial tissue changes, when other factors including the apparatus and technique remain stable. PMID- 2977797 TI - [Distribution patterns of hypertrophy at the apical level in patients with giant negative T waves: identification by magnetic resonance imaging]. AB - To clarify the distribution patterns of hypertrophy at the apical level in patients with giant negative T waves (GNT), ECG-gated magnetic resonance imaging (MRI) was performed in 10 patients with GNT and in five normal controls. End diastolic left ventricular short-axis images at the basal and apical levels were obtained in all subjects. Thicknesses of the septal, anterior, lateral and posterior walls at these two levels were measured and distribution of hypertrophied myocardium (more than or equal to 15 mm) at the apical level was examined. The ratio (R) of the maximal thickness at the apical level over that at the basal level was calculated. In normal subjects, the mean apical wall thickness was 8.7 +/- 1.9 mm. In the GNT group, the wall thickness was always greater than the mean value +3 SD of the normal control, and there were no differences among the four segmental walls. The hypertrophic portions at the apex were circumferential in three, septal-anterior-lateral in two, septal-anterior in two, septal in one, anterior in one and lateral in one. In patients with GNT, the average maximal thickness at the apical level was 19.3 +/- 3.2 mm; by location, four cases in the septum, four in the anterior wall and two in the lateral wall, and the average minimum thickness was 11.7 +/- 3.7 mm; all in the posterior wall. The R was more than 1.3 in nine patients with GNT and less than 1.0 in all normal subjects. In conclusion, there was a variety of patterns of apical hypertrophy, and the R greater than or equal to 1.3 was characteristic in patients with GNT. PMID- 2977798 TI - [Four cases of Fabry's disease mimicking hypertrophic cardiomyopathy]. AB - Four patients with Fabry's disease diagnosed by right ventricular endomyocardial biopsy had cardiac manifestations simulating hypertrophic cardiomyopathy (HCM). Case 1: A 51-year-old woman, whose elder sister had congestive heart failure, was hospitalized for exertional dyspnea and cardiomegaly. Her electrocardiogram (ECG) showed a short PQ interval (0.10 sec) and left ventricular hypertrophy. Her echocardiogram (Echo) showed moderate symmetrical hypertrophy of the left ventricle (IVST/PWT = 18 mm/17 mm). Case 2: A 32-year-old woman, whose elder sister had an abnormal ECG, was hospitalized for the ECG abnormalities consisting of a short PQ interval (0.10 sec) and ST-T changes in the left precordial leads. The Echo revealed mild symmetrical hypertrophy of the left ventricle (IVST = 13 mm, PWT = 13 mm). Case 3: A 44-year-old man was hospitalized for his ECG suggestive of left ventricular hypertrophy, and his Echo showed asymmetrical septal hypertrophy (ASH; IVST = 22 mm). Case 4: A 51-year-old man was hospitalized for his ECG showing high voltage in the left precordial leads, and his Echo showed ASH (IVST = 20 mm). The cardiac histopathological findings of these cases included cytoplasmic vacuolization by light microscopy, and electron dense deposits consisting of parallel or concentric lamellae with periodic spacing, suggesting Fabry's disease. The urinary glycolipids of Case 1 were increased biochemically; then the diagnosis of Fabry's disease was confirmed. Cardiac hypertrophy in Fabry's disease has many aspects, because the histopathological changes and clinical manifestations are determined by genetic factors. It was concluded that Fabry's disease may be concealed in some patients with the clinical diagnosis of HCM. PMID- 2977799 TI - Optimal timing for valve replacement in chronic aortic regurgitation: analysis based on the myocardial contractility and postoperative prognosis. AB - Left ventricular (LV) function was evaluated considering force-velocity and stress-shortening relationships in 14 asymptomatic (Group 1), 18 symptomatic patients (Group 2), and 53 normal subjects (Group 3) to determine the optimal time for valve replacement in patients with chronic aortic regurgitation (AR). Valve replacement was recommended for all Group 2 patients and for one patient in Group 1, who had sustained ventricular tachycardia. There was one operative death and five deaths remote from surgery; one patient in Group 1 died suddenly of undetermined cause, and four patients in Group 2 died of congestive heart failure (CHF). The LV end-systolic volume index (ESVI) was greater than 100 ml/m2 in the five patients whose death was unrelated to surgery (remote deaths). ESVI was less than 50 ml/m2 in all but two patients in Group 1, and more than 40 ml/m2 in all cases in Group 2. The index of preload, end-diastolic stress (sigma ed), was increased in Groups 1 and 2 as compared with Group 3. A significant positive correlation was observed between end-systolic stress (sigma es) and ESVI (r = 0.71, p less than 0.001) in patients with AR, and this linear line was not as steep as that of Group 3. Afterload (sigma es) and ejection fraction (EF) in Group 1 were within normal range, afterload was normal but EF was reduced in mildly symptomatic patients in Group 2, and severely symptomatic patients had markedly reduced EF and elevated afterload. There was a close correlation between ESVI and end-diastolic volume index (EDVI), and this was expressed as an exponential curve (Y = 21.69e0.006x, r = 0.88, p less than 0.001). This indicates that the rate of shortening of the muscle fiber deteriorates exponentially with enlargement of the ventricle. These observations suggest that in AR patients: 1) afterload and contractility of the ventricle remain normal in the majority of asymptomatic patients by means of compensatory hypertrophy and preload elevation (preload reserve), 2) deterioration of contractility seems to be the factor initiating CHF, and 3) progression of CHF is due to further deterioration of contractility in addition to elevation of afterload (afterload mismatch). It is concluded that careful observations are necessary when ESVI exceeds 50 ml/m2 in asymptomatic patients. Valve replacement is recommended when such patients develop symptoms of CHF, or either when EF falls to less than 50% or ESVI exceeds 100 ml/m2, even if patients remain asymptomatic. PMID- 2977800 TI - [Surgical indications of ischemic heart disease: a physician's viewpoint]. AB - Surgical indications for patients with ischemic heart disease are discussed from a physician's viewpoint. Among the subsets of this disease, surgical treatment is undoubtedly recommended for patients with serious complications following acute myocardial infarction (MI) such as cardiac rupture, interventricular septal perforation, acute severe mitral regurgitation and left ventricular mural thrombi complicated by systemic emboli, because these subsets have little or no response to medical treatment. Patients with left ventricular aneurysm are also surgical candidates, if they have refractory left heart failure or ventricular arrhythmias. Indications for coronary artery bypass surgery (CABS) for patients with angina pectoris are as yet controversial in Japan among physicians and surgeons, because the Japanese comply well with medications, and medically treated patients have favorable prognoses. Indication standards for CABS may consist of three components; the first, an essential condition of the institution where CABS is performed. It includes the physician's ability to precisely diagnose angina pectoris, availability of skilled surgeons and anesthesiologists for heart surgery, a well-organized system for patient management during and after CABS and an acceptable success rate. The second includes a clinical indication. It is considered valid that candidates for CABS should be basically patients unresponsive to medical treatment, in so far as there would be a risk such as perioperative MI or operative death. Angina pectoris is judged to be medically-resistant when chest pain at rest does not resolve, or an exercise capacity necessary for patients' social rehabilitation is not achievable, after the appropriate administration of antianginal agents including a combination of nitrates, calcium antagonists and beta-blockers. The last problem is an anatomical indication. Recent advances in percutaneous transluminal coronary angioplasty (PTCA), an effective therapeutic procedure for coronary revascularization, has greatly influenced indications for CABS, and it makes it difficult to strictly distinguish medical from surgical treatment. The purpose, clinical indications, and efficacy of PTCA are similar to those of CABS. However, patients' physical, emotional and socioeconomic burdens relative to PTCA are remarkably less than those of CABS. Thus, it seems reasonable that CABS should be recommended to patients who have contraindications to or high risks with PTCA, such as left main trunk lesions; whereas in general, there are no anatomical contraindications to CABS. PMID- 2977801 TI - The effect of pH on the calcium dependence of calcium accumulation in dog cardiac muscle sarcoplasmic reticulum. AB - Net Ca2+ accumulation in vesicles of dog cardiac sarcoplasmic reticulum (CSR) was evaluated at three different pHs: 6.0, 6.8 and 7.6. The Ca2+ sequestration by CSR depends on Ca2+ concentration and on pH values. The curves that show the relationship between Ca2+ accumulated by CSR and external Ca2+ concentrations were shifted with pH changes, both in the absence and in the presence of potassium oxalate. Considering the curve at pH 6.8 as reference, a lower Ca concentration was needed to obtain the half-maximal value in Ca sequestration under pH 7.6 (0.04 +/- 0.006 and 0.79 +/- 0.09 microM at pH 7.6 and 6.8, respectively). Opposite results were obtained under pH 6.0 (13.66 +/- 1.29 microM). Net calcium release during active accumulation of Ca2+ and Ca2+ efflux from passively 45Ca2+ loaded CSR microsomes were significantly higher at alkaline pH than at acidic pH. The results suggest that in CSR alkaline pH would promote the increase in the rates of both, Ca2+ release and active Ca2+ accumulation, while opposite effects would be expected under acidic pH. Therefore, pH changes may regulate both, the Ca2+ level upon which the SR Ca2+ pump works (permeability effect) and the sequestration rate of the Ca2+ pump (variation in the affinity for calcium). PMID- 2977802 TI - Significance of changes in plasma atrial natriuretic peptide in congestive heart failure. PMID- 2977803 TI - [A transient appearance of GOT-linked immunoglobulins]. PMID- 2977804 TI - [Study of multicentric reticulohistiocytosis on cultured histiocyte like cells]. PMID- 2977805 TI - Survey and observation of physically handicapped persons living at home due to intractable disease. PMID- 2977806 TI - [The influence of dementia and physical disability on the circadian rhythm of heart rate in the elderly]. PMID- 2977807 TI - [Estimation of myocardial sympathetic neuronal function with I-123 metaiodobenzylguanidine (MIBG) in patients with acute myocardial infarction undergoing percutaneous transluminal coronary thrombolysis (PTCR)]. PMID- 2977808 TI - [Percutaneous transluminal coronary angioplasty in 10 patients after A-C bypass]. PMID- 2977809 TI - [PTCA for completely occluded saphenous aortocoronary graft]. PMID- 2977810 TI - Abnormal in vitro differentiation of clonogenic B-cells in common acute lymphoblastic leukemia in complete remission. A marker for minimal residual disease? PMID- 2977811 TI - Can neo-adjuvant chemotherapy prevent residual tumors? PMID- 2977812 TI - [Structural characteristics of alkylated DNA in phage lambda]. PMID- 2977813 TI - [Isolation of strains of the hemorrhagic fever with renal syndrome virus in a cell culture]. PMID- 2977814 TI - [Combined immobilized cultures producing fibrinolytic proteinases]. AB - The authors obtained combined immobilised systems composed of a culture producing fibrinolytic proteinases and a stimulating strain. The optimal ratio between the two cultures in gel was selected at a high starting cell density. Highly stable immobilised cultures were produced by growing the cells in gel particles. The interrelationship of the partners was studied in the binary immobilised culture. The biosynthetic activity of the system fell down to the level of a monoculture when the cells of the stimulating strain were eliminated from gel. The producing and stimulating strains are at equilibrium in associative immobilised cultures obtained by growing the cells in gel, and Arthrobacter is not eliminated. The mechanism of biosynthesis stimulation in a combined immobilised culture has been studied. Apparently, the procedure of immobilisation and the action of a stimulating compound exert the synergistic effect. PMID- 2977815 TI - [Usefulness of ultrasonic evaluation of the ophthalmic artery branches by the Doppler method in the diagnosis of marked stenosis and occlusion of the internal carotid artery]. AB - Ninety-two patients with disturbances of cerebral circulation were subjected to contrast diagnostic methods and ultrasonographic examination by the method based on Doppler's phenomena. In ultrasonographic examination the blood flow in the internal carotid artery was assessed directly and indirectly (through the extraorbital branches of the ipsilateral ophthalmic arteries with compression test). The results of Doppler ultrasonography of 119 internal carotid arteries were compared with their angiographic images which showed normal state of these arteries, in 56 stenosis of the lumen over 70% in 14, occlusion of the artery in 49 (with occlusion of the initial part of the artery in 44 cases). Using the combined evaluation of the internal carotid and ophthalmic arteries a high agreement was obtained between the results of ultrasonography and contrast methods--96.3% for normal internal carotid arteries, 93.6% for arteries with major stenosis or occlusion, 90.9% for occlusion of the initial part of the internal carotid artery. It is worth stressing that in Doppler ultrasonography only a combination of absent blood flow in the artery and lack of Doppler's effect or reversed blood flow in the ipsilateral ophthalmic artery indicate unequivocally occlusion of the internal carotid artery. PMID- 2977816 TI - Atrial natriuretic peptide directly inhibits corticosteroid biosynthesis in human aldosterone-producing adenoma. AB - The effect of human atrial natriuretic peptide (hANP) on aldosterone, 18-OH cortisterone, corticosterone, deoxycorticosterone, and cortisol secretion was examined in isolated human aldosteronoma cells from five patients with primary aldosteronism. hANP exerted a nearly identical inhibitory action on basal secretion of each of these steroids and antagonized also the stimulating effects of ACTH, serotonin, metoclopramide and vasoactive intestinal polypeptide. The results suggest that in human aldosteronoma cells hANP may be a non-selective inhibitor of corticosteroid biosynthesis and that the site of inhibition of steroidogenesis may be localized to the early pathway of steroid biosynthesis. PMID- 2977817 TI - The Scandinavian multicenter photopatch study 1980-1985: final report. AB - At 16 different dermatology clinics in Scandinavia from 1980-1985, photopatch testing was performed on 1993 patients with suspected photodermatosis. The collective results are presented in this article. The most common cause of sun related dermatosis was polymorphic light eruption (PLE) (38%), while secondary aggravation of pre-existing skin diseases was established in 16% of the patient group. Photocontact dermatitis (11%) and contact dermatitis (10%) were responsible for 274 and 369 positive test reactions (respectively) on photopatch testing using the SPDRG standard series. Musk ambrette and para-aminobenzoic acid were the leading photosensitizers, while perfume mixture, balsam of Peru and lichen mixture were the most frequent causes of contact sensitivity. The principal photoallergens and contact allergens in the PLE, persistent light reaction and actinic reticuloid groups are discussed, together with the problems, risks and possible mechanisms of induction of photosensitization in these patients. The incidence, causes and diagnostic and therapeutic implications of secondary sunscreen sensitivity in these groups are also addressed. PMID- 2977818 TI - The relationship of the renin-angiotensin-aldosterone system to atrial natriuretic peptide and the natriuresis of volume expansion in diabetics with and without proteinuria. AB - Atrial natriuretic peptide (ANP) and the renin-angiotensin-aldosterone system are important regulatory hormones in sodium homeostasis. We have measured these hormones during volume expansion produced by water immersion in diabetic subjects without and with microalbuminuria or frank proteinuria and compared the response with normal controls. Diabetic subjects excreted about half the amount of sodium that was excreted by the normal subjects (39 vs 21 mmol) over 4 h. Diabetic subjects and normal ones showed a twofold rise in ANP during immersion and a marked suppression of both plasma renin activity and aldosterone. There was no difference in the hormonal response between diabetic and normal subjects or between those diabetic subjects with and those without incipient (microalbuminurics) or established nephropathy. PMID- 2977819 TI - Cerebral infarction and left ventricular mass: a clinical and echocardiographic study. AB - One hundred and two stroke patients were studied. Thirty-three (32 per cent) were hypertensive by the WHO criteria. Eighty-three (83 per cent) had cerebral infarction and three patients suffered from spontaneous intracerebral haemorrhage. The mean left ventricular mass was calculated from echocardiographic measurements and compared with that of controls. Neither cases nor controls had valvular or congenital heart disease, or disease processes that may be associated with myocardial infiltration. Mean left ventricular mass of all cases was significantly greater than that of controls (p less than 0.025) and that of the cases over the age of 50 years was also significantly greater than that of controls of the same age (p less than 0.02). The clinically normotensive cases had greater left ventricular mass than the normotensive controls (p less than 0.02). Meanwhile left ventricular mass in patients aged 50 and under was not significantly different from the appropriate control group (p greater than 0.2). These data indicate that the frequency of arterial hypertension among victims of cerebral infarction is greater than may be ascertained clinically particularly in those over 50. PMID- 2977820 TI - [The role of the staphylococcal infectious-allergic process in chronic marginal periodontal diseases, and specific therapeutic possibilities with staphylococcal vaccine. Preliminary note]. PMID- 2977821 TI - [Actualities concerning the pathogeny of reciprocal cracking noises in the temporomandibular articulations in the setting of the painful dysfunctional menisco-condyllary syndrome]. PMID- 2977823 TI - [The incidence of traumatic maxillo-facial lesions in the casuistics of the Bucco maxillo-facial Surgery Clinic from Bucharest]. PMID- 2977822 TI - [The temporomandibular articulation: internal disturbances and associated neuromuscular disorders]. PMID- 2977824 TI - [Adhesive resins for sealing of troughs and fossae]. PMID- 2977825 TI - [An epidemiological study of the contribution of intranatal risk factors to the frequency of dento-bucco-maxillary anomalies of children]. PMID- 2977828 TI - [Some aspects of the radiological assessment of the dento-periodontal support]. PMID- 2977827 TI - [A possible model for the interpretation of onset in chronic marginal periodontitis]. PMID- 2977826 TI - [The first stage of mixed dentition: an orthodontic approach]. PMID- 2977831 TI - [Late results following maxillary resections for malignant tumours]. PMID- 2977829 TI - [Aspects of the evolution of radiological investigative methods of the temporo mandibular articulations]. PMID- 2977830 TI - [Considerations on the cleaning of acrylic prostheses]. PMID- 2977832 TI - [Internal suspension in maxillary fractures]. PMID- 2977834 TI - [Magnetopressopuncture as a means for sedating children during stomatological treatment]. PMID- 2977833 TI - [Auriculotherapy-a method for inducing analgesia in the stomatological practice]. PMID- 2977836 TI - [General accidents in the dental office. Considerations of pathogenesis, prophylaxis and treatment]. PMID- 2977835 TI - [A method for the preparation of scale models of conjunct prosthetic devices made of thermally polimerized acrylate]. PMID- 2977839 TI - [Present stage in the treatment of bucco-maxillo-facial cancer]. PMID- 2977837 TI - [Allergic manifestations in the stomatological practice]. PMID- 2977838 TI - [Diagnostic problems in bucco-maxillo-facial cancer]. PMID- 2977840 TI - [The indications and limits of surgery in fractures of the facial mass]. PMID- 2977841 TI - [Plastic reconstructions after resections for labial malignant tumours]. PMID- 2977842 TI - [Tactical elements in the treatment of associated maxillo-facial and craniocerebral injuries]. PMID- 2977843 TI - [The treatment of fractures of the bones of the face. Experience in 8,346 cases]. PMID- 2977844 TI - [The study of glycoproteins from dental plaque]. PMID- 2977845 TI - [Structural and ultrastructural changes in the cerebral cortex, the hypothalamus, and of the adenohypophysis (TSH and FSH cells) in rabbits with experimental periodontitis ]. PMID- 2977846 TI - [Clinical and therapeutic results obtained in the treatment of affections of the buccal mucosa with an adhesive apiphytotherapeutic preparation, "Propovit MSK"]. PMID- 2977847 TI - [Pre-therapeutic stages of malignant tumours of the salivary glands]. PMID- 2977848 TI - [Branchiogenic carcinoma. Contributions to the definition of a controversial anatomo-clinical entity]. PMID- 2977850 TI - [The volemic balance in extensive bucco-maxillo-facial resections]. PMID- 2977849 TI - [Chondrosarcomas of the jaws]. PMID- 2977851 TI - [The importance of vertical redimensioning in the recovery of the occluding balance]. PMID- 2977852 TI - [Therapeutic principles for the resolving of dental trauma]. PMID- 2977854 TI - [A procedure for achieving the functional morphology of models]. PMID- 2977853 TI - [Experimental studies on marginal closure with three sealing agents]. PMID- 2977855 TI - [Options in working out a therapeutic approach in Down's syndrome cases]. PMID- 2977856 TI - Influence of thromboxane metabolism interference on glycemia regulation in rats. PMID- 2977857 TI - [Desmoid tumor: a review of the literature and report of 19 cases]. PMID- 2977858 TI - Assessment of gastric blood flow with laser Doppler flowmetry. AB - The use of laser Doppler flowmetry (LDF) to measure gastric blood flow was evaluated in the cat and man. The reproducibility of laser Doppler flowmetry recordings was studied in the feline stomach. In five cats flowmeter signals and venous outflow of the stomach were simultaneously recorded. The flowmeter recordings were made during operation and gastroscopy in 140 patients by means of two different (PF1-4-kHz and PF2-12-kHz) laser Doppler systems. The flowmeter recordings were highly reproducible during the cat experiments, with a coefficient of variation varying between 4% and 13%. Angulation of the probe within 60-120 degrees against the tissue under study did not affect the flowmeter signal. Pressure of the probe against the studied tissue attenuated the flowmeter signal 42 +/- 13% (n = 10). A significant correlation coefficient (r = 0.76; p = 0.01; n = 30) was obtained between flowmeter signal and venous outflow of the stomach. In man the PF2-12-kHz system yielded a higher flowmeter signal (14.5 +/- 6.9 V; 32 patients) than the PF1-4-kHz system (7.9 +/- 2.5 V; 108 patients). Flow dimensions were calculated by using results previously obtained in the intestines. The estimated blood flow values amounted to 31 +/- 10 and 57 +/- 27 ml min-1 100 g-1 for the PF1-4-kHz and PF2-12-kHz systems, respectively. In conclusion, laser Doppler flowmeter is a promising technique for studying gastric perfusion during operation and endoscopy. Motion artifacts are the major drawback of the present laser Doppler systems. PMID- 2977859 TI - [Involvement of rat locus coeruleus in the analgesic effect induced by monosodium glutamate injection into the hypothalamic arcuate nucleus area]. PMID- 2977860 TI - [Effects of atriopeptin III on isolated perfused rat heart]. PMID- 2977861 TI - Distribution of interleukin-2 receptors in activated T-lymphocyte subsets in vitro. PMID- 2977862 TI - [Periodontal disease in diabetes mellitus]. PMID- 2977863 TI - [Side effects of immunomodulators and countermeasures]. PMID- 2977864 TI - [Laparoscopy in the diagnosis of acute appendicitis in children]. AB - Based upon an analysis of results of 1210 operations and 160 laparoscopies in children for "acute appendicitis" the authors make a conclusion that about 20% of children were subjected to unnecessary appendectomies. Laparoscopy allows to make the correct diagnosis in the absolute majority of children with obscure clinical picture of acute appendicitis and to choose a rational medical treatment. PMID- 2977865 TI - [Spring retractors]. PMID- 2977866 TI - [Surgical treatment of endemic nodular goiter]. PMID- 2977867 TI - Perigraft reaction: incompatibility of synthetic vascular grafts? New aspects on clinical manifestation, pathogenesis, and therapy. PMID- 2977868 TI - Percutaneous transluminal angioplasty: the surgeon's role. PMID- 2977869 TI - [Aortic isthmus stenoses--dilatation in adulthood. A German cooperative study]. AB - An analysis of percutaneous transluminal angioplasty of the coarctation of the aorta in adults was evaluated in a cooperative study of the German Working Group of Angioplasty of the German Society of Cardiology. Dilation was performed in 18 patients with a mean age of 26 years (14-49 years). The success rate (gradient less than or equal to 20 mm Hg) was 78% regarding peak to peak gradient, 89% regarding mean gradient. The peak-to-peak gradient decreased from 82 +/- 16 mm Hg to 18 +/- 11 mm Hg. The diameter of the aortic isthmus increased from 0.7 +/- 0.3 cm to 1.3 +/- 0.4 cm (p less than 0.01). After six months only one restenosis occurred. The peak-to-peak gradient measured 10 +/- 12 mm Hg, the diameter 1.4 +/ 0.5 cm. In two patients a balloon rupture occurred without rupture-related complications. No patients died, no cross paralysis or aortic rupture occurred. In three of seven patients with trans-esophageal echocardiographic monitoring a small intimal flap was found; in one patient a media dissection occurred leading to a 15-min period of chest pain and spontaneous healing. In another patient successful dilation was controlled by acute control aortography and computer tomography. After discharge severe chest pain developed. A subtraction angiography of the aorta was negative. Six months later a biplane aortography of the distal thoracic aorta confirmed the diagnosis of aortic dissection type III DeBakey, previously diagnosed by transesophageal echocardiography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977871 TI - Intervertebral disc and tendon: quantitative ultrastructure and differentiation of their collagen fibrils. PMID- 2977870 TI - [Pseudostabilization of unstable angina pectoris: disappearance of symptoms in persistence of silent myocardial ischemia]. AB - We studied a patient hospitalized with unstable angina pectoris; ST-segment analysis during Holter ECG revealed several silent ischemic attacks despite complete disappearance of anginal symptoms under medical treatment. Prior to cardiac catheterization the patient went into acute myocardial infarction. Immediate intravenous thrombolysis and subsequent angioplasty of a high-grade stenosis abolished the ischemic events. ST-segment analysis during Holter ECG offers a method to detect ischemic events despite the disappearance of anginal symptoms in the clinical course of unstable angina pectoris. This technique might therefore identify patients with unstable angina pectoris at higher risk for further cardiac events. PMID- 2977872 TI - [The functional morphology of the adrenal cortex after experimental stress]. PMID- 2977873 TI - [Dog bite wounds: characteristics and therapeutic principles]. AB - Dog bite wounds are a common problem and children are the age group at highest risk. Wounds are often multiple and generally involve the face or the limbs. Injuries include punctures, crushes, lacerations and avulsions of flaps. The well know prevalence of facial injuries can pose some difficult reconstructive problems. Infections are not rare even in well-treated cases. The Authors report the last ten cases which they have treated and on the basis of their experience and of the literature suggest guidelines for effective treatment. A prophylactic antibiotic therapy and tetanus immunization are the basis of rational medical therapy. Wounds should be immediately sutured after careful debridement and through irrigation, unless they are heavily contaminated or located on the hand. PMID- 2977874 TI - [Surgery of the saphenous-popliteal junction in the treatment of initial insufficiency of the small saphenous vein]. AB - Insufficiency of the sapheno-popliteal junction may be the first sign of varicose disease, and represents one of several ways that lower limb-venous insufficiency uses to present itself. Doppler ultrasound is particularly helpful in diagnosing this often subclinical insufficiency. Based on satisfactory results in a series of 18 patients treated between 1985 and 1988 (mean follow-up 1.5 years), the authors suggest treating sapheno-popliteal junction insufficiency by means of crossectomy, under local anesthesia, and on an out-patient basis. When surgical treatment is performed during the first stages of the disease, the consequent irreversible damages to the lesser saphenous vein due to stasis are avoided, and thereby, a true varicose state is avoided. PMID- 2977876 TI - [Transcranial pulsed Doppler in the diagnosis of cerebrovascular pathology: methodology, indications and limitations]. AB - Doppler analysis of flow in intracranial arteries is now possible using a 2 MHz probe, allowing sufficient penetration of bone to obtain signals noninvasively and to measure rate of flow in middle (MCA), anterior (ACA) and posterior (PCA) cerebral arteries and in basilar trunk. Practical applications of this new method are numerous in cerebrovascular disease and, despite certain limitations due to anatomic factors, transcranial Doppler may identify stenosis of the intracranial arteries and study the value of collateral flow across the anterior circle of Willis in patients with extracranial carotid artery stenosis. PMID- 2977875 TI - [Current trends in the therapy of C.I.N. (cervical intraepithelial neoplasia)]. AB - The Authors examine the epidemiological data on cervical intraepithelial neoplasia (C.I.N.) and review the various therapeutic methods available for it's treatment, paying special attention to the outpatient treatment of these precancerous portio lesions. PMID- 2977877 TI - [Analysis and evaluation of blood alcohol values found in victims of fatal traffic accidents in the Parma region (1982-1986)]. AB - The Authors examined the relationship between blood alcohol levels and traffic accident incidence in 340 cases which came under the observation of the Institute of Forensic Medicine from 1982 to 1986. Positive blood alcohol (31.2% of cases) were correlated to age, sex, type of vehicle driven, day and hour of the accident and to influence upon behaviour (greater than or equal to 0.50 g%) and to drunkenness (greater than or equal to 1.50 g%). Positive blood alcohol resulted to be correlated to, in the following order, passengers, drivers, pedestrians, motorcyclists and cyclists. The incidence was also related to evening and night hours, and to the days Saturday, Sunday and Monday. Our results, even if not completely superimposable, from a statistical point of view, do permit a comparison between them and those found in the Literature regarding the casuistry of other forensic anatomo-pathologic studies that define the risk factors related to alcohol consumption and traffic accident risk. PMID- 2977878 TI - [A different interpretation of the functional role of uterine fixation during natural delivery]. AB - The Author suggests that physiologic delivery is triggered by the synergism of muscular and hydraulic forces. The first to be put into motion are the muscular contractions of the uterus, diaphragm and abdominal wall muscles; the second follow the first and cause a type of hydraulic press phenomenon that, by means of amniotic fluid, is spread to the pelvic basin during the phase in which the fetus is propelled and are only subjected to the muscular forces from myometrium contractions. Therefore, it is inaccurate to believe that these maintain all the forces of delivery. In fact, the hydraulic component of the forces of delivery, which is accepted as being the most important in terms of efficacy, doesn't rely on fixation by the uterus but rather relies on the bony surface of the pelvic basin. The Author believes that it is necessary to re-examine some of the traditional aspects of interpreting delivery because in spite of the rapid progress towards a complete understanding, the study of obstetrics has accepted without criticism and inadvertently assimilated concepts that originate and can be credited to the time of Hippocrates. PMID- 2977879 TI - [Systems assisting decisions in clinical medicine]. AB - There are numerous systems and computerized procedures which have been developed and tested as a guide to clinical decisions. The methods used in the development of these systems can be divided into two large groups: mathematical and logical. None of these can be considered as better than the other in absolute, but the efficiency of each system depends on that problem is to be described and solved. The mathematical methods seem more suitable in the description of biological systems and for the selection of groups which have discriminant and sufficiently defined characteristics. Meanwhile the logical methods are better in the description and the formalization of more complicated clinical problems, characterized by uncertainty and availability of more or less numerous alternatives. It is foreseen that in the next years the systems for aided decision making will be programmed making use of methods belonging to both categories, and particularly, the expert systems will be planned using both artificial intelligence techniques and mathematical and statistical methods. The increasing frequency in the use of such systems in clinical practice will cause a reevaluation and a checking of most aspects of medical proceedings, as seen by the new methodologies or the traditional methods whose use could be driven by computer. PMID- 2977880 TI - [Study on health status and oral hygiene of children at a primary school. Health status and oral hygiene in schoolchildren]. AB - The Authors refer about a research made in a primary school after checking mouth sickness in school boys. The research concerns three periods: 1st. medicaL examination, 2 nd. instructions about a correct oral hygiene, 3 rd. anew inspection. The conclusions they made, are the following: now only the children know what they must do for their teeth in order to avoid mouth sickness and how to prevent then, but also their parents and often their teachers are not sensitive to this problem. So it is necessary that the prevention work begun by the dentist, will be carried on by parents and by teachers. PMID- 2977881 TI - [Prenatal development of muscle spindles in pig extraocular muscles and its significance]. PMID- 2977882 TI - Plasma atrial natriuretic peptide (ANP) and maternal hemodynamic changes during normal pregnancy. AB - Peripheral venous plasma concentrations of immunoreactive atrial natriuretic peptide (irANP) were studied longitudinally in 12 women at the 12th, 24th, and 36th week of pregnancy as well as 3-5 days and 3 months post partum. Serial measurements of maternal hemodynamics were performed simultaneously with blood sampling for irANP determination. With advancing pregnancy there were significant increases (p less than 0.001) in cardiac output, stroke volume and heart rate, while total peripheral vascular resistance decreased (p less than 0.001). All these changes were normalized 3 months post partum. Plasma irANP increased (p less than 0.05) from 23.2 +/- 1.3 pM/l at week 12 to 25.9 +/- 1.5 pM/l at week 36 of pregnancy, and fell significantly (p less than 0.01) to 20.5 +/- 1.1 pM/l 3 months post partum. Changes in plasma irANP appear to be related to changes in maternal central hemodynamics. The changes in ANP release probably represent one of several mechanisms that maintain circulatory and volume homeostasis during normal pregnancy. PMID- 2977883 TI - Serial measurements of plasma renin activity, aldosterone and cortisol during percutaneous transluminal angioplasty of the renal artery in man. AB - Fifteen patients (eight men, seven women) with hypertension and renal artery stenosis underwent dilation of the stenosis by percutaneous transluminal renal angioplasty (PTRA). During and shortly after this treatment the effects on the renin-angiotensin-aldosterone system and blood pressure were studied. Plasma renin activity (PRA) was measured in peripheral blood and in renal venous blood during the PTRA. PRA increased in peripheral blood during PTRA as a result of an immediate significant rise in renal venous plasma renin activity by 132 +/- 134% (P less than 0.01) on the dilated side. PRA in the contralateral renal vein was close to that in peripheral blood. Within 10 min after PTRA there was a significant increase in serum aldosterone from 439 +/- 343 to 774 +/- 635 pmol 1 1 (P less than 0.025), while serum cortisol remained unchanged. The aldosterone increase was most probably mediated by angiotensin II. Systolic and diastolic blood pressures were unchanged during PTRA in spite of renin and aldosterone increases, suggesting that antihypertensive factors counteract the pressor effects of a physiologically relevant increase in PRA. PMID- 2977884 TI - Ultrastructural changes of synapses in supraoptic nucleus of hedgehog during hibernation. PMID- 2977886 TI - Probability of paternity: deceased accused man or mother; twins. PMID- 2977885 TI - Polymorphic systems in North Moravian Region. PMID- 2977887 TI - Does circadian variation of "cathepsin B-like" activity exist in the serum of patients with lung cancer? PMID- 2977888 TI - Healing of experimental laser wound in liver and spleen. PMID- 2977889 TI - Healing of experimental laser wound in the skin and auricle. PMID- 2977890 TI - Healing of experimental laser wound in striated muscle and adipose tissue. PMID- 2977891 TI - Early gastric carcinoma. PMID- 2977892 TI - Retrospective study of gastric cancer on the basis of Lauren's classification criteria. PMID- 2977894 TI - [Histologic findings in chronic inflammation of the maxillary sinuses]. PMID- 2977893 TI - Morphological correlates between bone marrow findings in patients with lymphocytic ML (CCL and immunocytoma) and clinical staging. PMID- 2977895 TI - Temperature-dependent dimorphism and growth rate of Trichophyton equinum mutants. PMID- 2977896 TI - Adsorption of alpha-amylase (EC 3.2.1.1) and lipase (EC 3.1.1.3) before and after substitution therapy in exocrine pancreatic insufficiency. PMID- 2977897 TI - Colicinogeny in patients after cholecystectomy. PMID- 2977898 TI - [Lyme disease]. PMID- 2977899 TI - Cytogenetic findings in 54 patients with polycythemia vera and their prognostic value. PMID- 2977900 TI - The seasonal dependence of magnesium and zinc in blood in young sportsmen. PMID- 2977901 TI - Innovation of the antiepileptic therapy with magnesium lactate. PMID- 2977902 TI - Our experience with amiodarone administration to patients with paroxysmal atrial fibrillation. PMID- 2977904 TI - Lactation-inhibiting and prolactin-lowering effect of lisuride and transdihydrolisuride. A comparative study. PMID- 2977903 TI - Terguride in the treatment of prolactin-induced luteal insufficiency. PMID- 2977905 TI - [The gastroesophageal junction. Present views of its function and our experimental studies]. PMID- 2977906 TI - [Hemodilution in general surgery]. PMID- 2977907 TI - Perioperation electrostimulation of the anal sphincters. PMID- 2977908 TI - Postirradiation rectovaginal fistula and its surgical management. PMID- 2977909 TI - Application of immunotherapy in proctosurgery. PMID- 2977910 TI - Some notes on the incidence of nosocomial infections at the department of surgery. PMID- 2977911 TI - Psychopharmacologic management of the stress in pedodontic practice. PMID- 2977912 TI - The anatomy of soft tissues of the lower vestibule in relationship to periodontal tissues of the lower incisor teeth. Epidemiological study. PMID- 2977913 TI - Contribution to diagnosis and treatment of pulpal-periodontal lesion from the standpoint of conservative stomatology. PMID- 2977914 TI - [Repair of removable partial dentures with metal skeletons]. PMID- 2977915 TI - Physiological serum vitamin E level and age. PMID- 2977916 TI - [Forensic medical and dental problems of identification of victims of mining disasters]. PMID- 2977917 TI - Chromatic temperature characteristics of thermographic sheets applied in actual Czechoslovak medical practice. PMID- 2977918 TI - Item intercorrelation in the multiple-choice test. PMID- 2977919 TI - Auxotrophic mutants of ovicidal fungus Penicillium lanosum. PMID- 2977920 TI - A contribution to the prenatal development of the lymphatic tissue of the parotid gland. PMID- 2977921 TI - Biotransformation of demecolcine by rat liver microsomes. PMID- 2977922 TI - Quantitative morphometric evaluation of changes in the small intestine mucosa of laboratory rats after gamma and mixed neutron and gamma irradiation. PMID- 2977923 TI - Enzyme cytochemistry of the rat plexus chorioideus after supralethal gamma irradiation. PMID- 2977924 TI - Reaction of some enzymes in nucleus supraopticus of rats irradiated with 300 Gy of gamma radiation. PMID- 2977926 TI - [Meosix--a simple method for the semiquantitative evaluation of enzyme activity in histochemistry]. PMID- 2977925 TI - [Topochemistry of hydrolases in the intestines of human embryos and fetuses]. PMID- 2977927 TI - Myelinated neurons in the central nervous system of the hedgehog. PMID- 2977928 TI - Kidney disease in KK mice: effect of glyburide. PMID- 2977929 TI - Some characteristics of chemically-induced beta-cell lesions as possible contributing factors in diabetes. PMID- 2977930 TI - Prevention of diabetes in the BB rat by injection of major histocompatibility complex- (MHC-) compatible bone marrow cells. PMID- 2977931 TI - Chemically-initiated hyperglycemia: streptozotocin-specific T lymphocyte reactions. PMID- 2977932 TI - [Non-reusable regulators of perfusion]. PMID- 2977933 TI - [Epidemiology of mental retardation. Study of a dependent population of a telephone company]. PMID- 2977934 TI - Left ventricular structural and functional changes in young men at increased risk of developing essential hypertension. Assessment by echocardiography. AB - In order to evaluate if left ventricular structural changes are present in young subjects with increased risk of developing essential hypertension, we have investigated young normotensive and borderline hypertensives with and without hypertensive parents. Our results showed, that left ventricular hypertrophy (LVH) was found in these risk subjects and that LVH was associated to borderline hypertension. PMID- 2977935 TI - Serum dehydroepiandrosterone sulphate and pubertal development in Chinese girls. AB - Serum dehydroepiandrosterone sulphate (DHEAS), estradiol (E2), luteinizing hormone (LH) and follicle stimulating hormone (FSH) were measured by radioimmunoassay in 170 healthy school girls from 7-16 years old. Blood samples from postmenarcheal girls were taken on days 6-10 of the menstrual cycle. Bone age, height, weight, breast and pubic hair development were also investigated. Of the four hormones measured, serum DHEAS displayed the earliest elevation, at 7 years and upwards. A significant DHEAS increment at chronological age 13 years was observed. Girls with earlier menarche had higher DHEAS level than girls having later menarche. Serum DHEAS levels also correlated with bone age, height, weight, subcutaneous fat and pubertal stages. The results suggest that adrenal androgen might be involved in the initiation of puberty and female maturation. PMID- 2977936 TI - Maternal age in trisomy. AB - By comparison with a more general theory, data on trisomy in live births, amniocenteses, and spontaneous abortions by year of maternal age are shown to fit a logistic augmented by a proportion independent of maternal age. The frequency of trisomy increases monotonically, with no discrepancy at extremely low or high maternal age. Trisomy 16 is exceptional in that all cases appear to be age dependent. For groups A, B, and C most trisomies arise by a process independent of maternal age. A small proportion of these trisomies and about half of trisomies for smaller chromosomes (excluding trisomies 16 and perhaps 22) originate by some process dependent on maternal (but not paternal) age and therefore presumably independent of crossingover, which in the female takes place before birth. PMID- 2977938 TI - [Atherosclerosis and its complications as a cause of hospital mortality in the Tashkent population in 1982-1986]. AB - Atherosclerosis and coronary disease incidence rate specific for age, sex and nationality has been estimated at 7108 autopsies performed at city hospital in 1982-1986. The analysis suggested this rate to be on the increase, being 2.4% for 1920s, 29.3% for 1977-1981 and 39.5% for 1982-1986. Localization of arterial lesions responsible for a primary disease varies. Coronary heart disease ranks first occurring in 67.9% of cases. CVD follows occurring in 22%. Atherosclerosis death rate for the inpatients when adjusted for sex is age-specific. Though atherosclerosis is more frequently detected in native males aged 45-54 and females aged 45-49 compared to newcomers, the latter outnumber the natives by cases of atherosclerosis in the rest groups. PMID- 2977937 TI - Proline-rich polypeptide (PRP) from ovine colostrum. Bi-directional modulation of binding of peanut agglutinin, resistance to hydrocortisone, and helper activity in murine thymocytes. AB - A proline-rich polypeptide (PRP) isolated from ovine colostrum has a regulatory effect on the immune response. The present report demonstrates that the polypeptide can cause bi-directional modulation of surface markers and function of murine thymocytes. PRP is able to reduce binding of peanut agglutinin (PNA) to murine PNA+ thymocytes and to increase the binding of PNA to PNA- cells. This transition of binding ability can be reversed by a second treatment of cells with PRP. PRP is also able to transform cortisone-resistant thymocytes into cortisone sensitive, and vice versa. Helper cells induced by PRP from murine glass nonadherent thymocytes (PNA+) showed suppressor activity after the second treatment with PRP. The observed changes were occurring concomitantly, i.e. changes in binding of PNA were accompanied by changes in resistance to cortisone and in expression of helper or suppressor activity. Bi-directional effects of PRP on PNA-binding ability, sensitivity to hydrocortisone, and helper-suppressor function, makes this polypeptide unique among immuno-modulators known. PMID- 2977939 TI - Fibre optic laser Doppler measurement of intravascular velocity. PMID- 2977940 TI - Tophaceous gout of the axial skeleton. AB - A 72-year-old female with a recent episode of podagra, without peripheral tophi, presented with nocturnal back pain and a destructive lesion of the right lamina of L4. A tophus was excised at laminectomy. Evidence of an axial skeletal tophus causing an irritative radiculopathy via mass effect is presented. PMID- 2977941 TI - Familial restrictive cardiomyopathy. AB - The cases of a father and daughter with idiopathic restrictive cardiomyopathy are described. In contrast to other forms of cardiomyopathy, this type is rarely familial. PMID- 2977942 TI - Adjuvant cytotoxic chemotherapy for colorectal cancer: the present position. PMID- 2977943 TI - Treatment of alopecia areata with diphencyprone. PMID- 2977944 TI - Maternal mortality in Rangoon Burma. PMID- 2977945 TI - Value of laparoscopy in the diagnosis of hepatic and peritoneal diseases. PMID- 2977946 TI - Lipid peroxidation and retinal phototoxic degeneration. PMID- 2977947 TI - Roles of oxygen and oxygen substitutes in DNA sugar damage by antitumor antibiotics. PMID- 2977948 TI - Studies on recombination within the mouse H-2 complex: IV. Characterization of new recombinant haplotypes H-2t7, H-2t8, H-2as2, and H-2as3. AB - Four new intra-H-2 recombinants were characterized serologically and functionally. In two of these recombinants, B10.ASR1 and B10.ASR7, crossing over occurred between the A alpha and E alpha subregions, very probably in E beta since most intra-I region recombinants thus far investigated at the DNA level appear to involve recombination within the E beta gene. In the other two recombinants, B10.ASR2 and B10.ARS8, crossing over occurred between the S and D subregions. B10.ASR2 and B10.ASR8, crossing over occurred between the S and D subregions. B10.ASR7, which is serologically indistinguishable from B10.BASR1 and B10.S(8R), slightly stimulates and strongly responds to both of these strains in MLR. The probable location of the B10.S(8R) stimulatory product is E beta. The H 2 composition of B10.ASR1 is closest to that of B10.S(9R) and B10.HTT; therefore, precise definition of the cross-over point at the DNA level will be of particular interest. PMID- 2977950 TI - [Esthetic inlays of high strength bis/GMA resin]. PMID- 2977949 TI - "Tolerization" of human T-helper cell clones by chronic exposure to alloantigen: culture conditions dictate autocrine proliferative status but not acquisition of cytotoxic potential and suppressor-induction capacity. AB - Induction of clonal anergy in T-helper (Th) cells may have a role in regulating immune responses. A model system for studying Th cell tolerization at the clonal level in vitro could be useful for investigating the mechanisms involved. Accordingly, alloreactive helper cells were maintained in culture with interleukin 2 (IL 2) by intermittent stimulation with specific antigen. Regardless of the frequency of antigen stimulation, clones of age less than ca. 35 population doublings (PD) were found to undergo antigen-specific autocrine clonal expansion in the absence of exogenous IL 2. Such young clones (designated as phase I) could therefore not be "tolerized" by frequent exposure to antigen. In contrast, most clones of age greater than ca. 35 PD could be tolerized by frequent exposure to antigen (designated as phase II clones). Their autocrine proliferation was then blocked, although they still recognized antigen specifically as shown by their retained ability to secrete interferon-gamma (IFN gamma) and granulocyte-macrophage colony stimulating factor (GM-CSF). The mechanism of response failure involved both an inability to upregulate IL 2 receptors in the absence of exogenous IL 2, as well as an inability to secrete IL 2. These defects were not overcome by stimulation with mitogens or calcium ionophore and phorbol esther in place of alloantigen. T-cell receptor, alpha, beta, and gamma-chain gene rearrangements remained identical in phase I and phase II clones. Tolerization of phase II clones could be avoided by increasing the period between antigen exposures. Despite this, whether or not phase II cells were capable of autocrine proliferation, they were found to have acquired the novel function of inducing suppressive activity in fresh lymphocytes. Suppressor induction was blocked by the broadly reactive MHC class II-specific monoclonal antibody (moAb) TU39, but not by moAb preferentially reacting only with HLA-DR, DQ, or DP. Sequential immunoprecipitation on T-cell clones showed the presence of a putative non-DR, DQ, DP, TU39+ molecule on phase II clones. However, this molecule was also found on phase I clones. The nature of the TU39-blockable suppressor-inducing determinant present on phase II but not on (most) phase I clones thus remains to be clarified. In addition to suppressor-induction activity, phase II clones also acquired lytic potential as measured in a lectin approximation system. Cytotoxic (CTX) potential was also not influenced by the frequency of antigenic stimulation and could be viewed as a constitutive modulation of clonal function.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2977951 TI - [The advantages of conscious sedation in care of the handicapped]. PMID- 2977952 TI - Rapid kinetic characterization of active transport and passive release of calcium in vesicular fragments of longitudinal and junctional sarcoplasmic reticulum. AB - 1. Vesicular fragments of longitudinal and cisternal-junctional sarcoplasmic reticulum can be obtained, selectively, from skeletal muscle. 2. Longitudinal sarcoplasmic reticulum vesicles have a high content of Ca2+-ATPase, and provide the best system for characterization of the calcium pump. Equilibrium and transient state measurements provide equilibrium and kinetic constants for the partial reactions of the catalytic and transport cycle, including binding and translocation steps. Free energy estimates are then obtained based on an explicit mechanism, rather than simply on a transmembrane gradient (which is independent of the reaction mechanism). Kinetic evidence for a sequential pattern of calcium binding and translocation is presented. 3. The cisternal-junctional membrane contains (in addition to Ca2+-ATPase) calsequestrin and a large protein (MW greater than or equal to 350,000 Da) which corresponds to the junctional processes ("feet") and binds ryanodine. This protein is involved in rapid release of Ca2+ from loaded vesicles. PMID- 2977953 TI - Evolutionary conservation of laminin-binding proteins. AB - 1. The virulence of pathogens and metastatic capacity of cancer cells seems to correlate with the ability to adhere to cells and/or to basement membrane components. A key feature of this mechanism is the expression of specific receptors for the basement membrane protein laminin. Three different receptors have been already described in cells phylogenetically very distant, such as human white blood cells, Trichomonas vaginalis and Staphylococcus aureus, all recognizing laminin with the same range of affinity. 2. We have shown that laminin, which is also found in the circulation, enhances phagocytosis of S. aureus by macrophages in a species-specific fashion. Also, monoclonal antibodies (MAb) raised against the bacterial receptor inhibit the phagocytic enhancement mediated by laminin and recognize laminin-binding proteins in unicellular parasites and mammalian cells. The same Mab 1.H12 elutes a 52-kDa protein from bacterial extracts and a 67-kDa band from cancer cell extracts. Since the MAb is a monospecific reagent, results with 1.H12 strongly suggest an evolutionary conservation of the binding site of phylogenetically different laminin receptors. PMID- 2977955 TI - The elderly, disabled and handicapped adult burned through abuse and neglect. AB - A retrospective review of 1152 acute burns treated at the University of Michigan Burn Center during the past 5.5 years identified 26 adult patients in whom abuse and neglect was suspected. These patients, 12 females and 14 males, had an average age of 42 years and average total body area burn (TBA) of 18 per cent. Eight of these cases were in situations in which the question of inflicted injury was raised and 18 were in situations of neglect and/or improper supervision. Seven (27 per cent) died from their injury. All of these adults were either very old, mentally impaired, and/or physically handicapped. Twenty-three were burned in health care facilities or institutions. Adult abuse in the elderly, handicapped and disabled population is similar to that found in child abuse and includes characteristic burn patterns, a story that does not fit the injury, and a delay in seeking medical care. An increased awareness of the problem is necessary for improved diagnosis and management of these patients. PMID- 2977954 TI - Post-partum psychosis in adult GM2 gangliosidosis. A case report. AB - Adult hexosaminidase A deficiency is a form of GM2 gangliosidosis with autosomal recessive inheritance. Only 35 cases (mostly among Ashkenazic Jews) have been reported worldwide. Symptoms include, in a third of the cases, psychosis. A 27 year-old sufferer with no prior psychiatric history, developed a post-partum psychosis, with affective and hebephrenic components, 3 days following her first delivery. She responded to lithium within 10 days of initiating treatment; the full episode lasted 1 month. We conclude that lithium is the preferred treatment for psychosis in such adult patients, especially in light of possible long-term neurological deterioration caused by phenothiazines. Ashkenazic Jews with atypical neurological syndromes presenting with psychosis should be tested for hexosaminidase A deficiency. PMID- 2977956 TI - Skin injury following contact with a complex amine. AB - Eight patients with skin damage due to contact with an amine are described. The material is used in the potash industry under the commercial name of Armine to prevent bosselation. Although it is considered to be a strong base, no deep burns were encountered. The reaction to this material initially resembled a dermatitis; later the wound developed the typical appearance of a burn. In addition to the common conservative treatment, antihistamine drugs were used. When a true allergic reaction was suspected steroids were added to the regimen. PMID- 2977957 TI - [Jean Brachet (1909-1988)]. PMID- 2977958 TI - [Changes in corticosterone and aldosterone concentrations in various tissues of Xenopus laevis tadpoles during the metamorphosis]. AB - During the metamorphosis of Xenopus laevis tadpoles, tissue concentrations of corticosterone and aldosterone did not change significantly in forelegs and hindlegs; they increased in tail, liver, skin and intestine. The rise of corticosteroid concentrations appeared in tissues which were deeply transformed during the first part of the climax, when plasma levels of corticosteroids also increased. Highest tissue levels, attained at the mid-climax, were maintained at least until the end of metamorphosis although plasma concentrations of two steroids were then abruptly fallen. Tissues able to retain corticosteroids reacted as Vertebrate "target tissues" and transformations which took place in them could be dependent, at least partially, on corticosteroids. PMID- 2977959 TI - [Effect of retinoic acid on the differentiation and growth of murine mastocyte precursors]. AB - In cultures of normal mouse hematopoietic cells containing Interleukin-3 develop cells with many features of mast cells. These cells seem heterogeneous with respect to morphological and biochemical examination. Nevertheless, most of the cells show many granules and a low ability to self-renew. In the present report we describe the development of a blastic cell population, termed mastoblasts, when normal mouse hematopoietic cells are exposed continuously to retinoic acid (RA: 10(-6) to 10(-5) M/l). Using H*3-thymidine incorporation, cell cycle measurement and protein content by flow cytometry, transmission and scanning electron microscopy, we show that these cells seem to be of mast cell lineage but with a high self-renewing capability. So, RA is able to inhibit mast cell differentiation and to provide us a "mastoblastic" population which could be used as a model to study mast cell differentiation. PMID- 2977960 TI - [Blood polymorphism in the study of isolated communities]. AB - Isolated communities offer a unique opportunity for the study of biological and social consequences of consanguinity and migration. The studies of genetic polymorphisms have contributed greatly, not only to knowledge of the genetic constitution of a given individual and population, but also to clarify either relationship between structure and function of polymorphic traits or the susceptibility to multifactorial diseases, in which interaction between the gene and environment cannot be ignored. For over 25 years, we have investigated the effect of consanguinity and genetic polymorphisms in 9 isolated communities in Western Japan. We reported here different values of gene frequency for each polymorphic trait, compared with the neighboring communities and described how we applied these data to clarification of the genetic constitution of isolated communities as well as of genetic susceptibility to some diseases. PMID- 2977961 TI - [Effect of temperature on the expression of bobbed mutations in Drosophila melanogaster]. AB - The expression of two bobbed mutations on the X chromosome was studied at two temperatures 25 degrees C and 18 degrees C (larval developmental time, viability, phenotype of adult...). Results showed that the wm4bb mutation is thermosensitive. Some hypothesis are expressed to explain this phenomenon. PMID- 2977962 TI - [A technic for selection of antidotes to an acute lethal poisoning]. AB - The dose of a toxic substance which is lethal to the rat within 15 min (LD100.15 min) and its specific antidotes are defined. Atropine, diazepam and nimodipine in sequential administration are effective antidotes to a LD100.15 min (1.5 mg/kg) of paraoxon, an anticholinesterase. PMID- 2977963 TI - [Effects of biologically active peptides on the giant neurons of the giant African snail, Achatina fulica Ferussac]. AB - Effects of neuropeptides proposed as neurotransmitters of mammals and invertebrates and those of natural toxins, etc., on the identifiable giant neurones of an African giant snail (Achatina fulica Ferussac) were examined. Oxytocin had a tendency to show the excitatory effects; this substance was excitatory on five neurones and inhibitory on one neurone. A substance related to oxytocin, vasotocin, also showed excitatory effects on the three neurones. On the other hand, FMR Famide was inhibitory on six neurones. Proctolin had excitatory effects on only one neurone. PMID- 2977964 TI - [Mechanism of tumor promotion]. AB - Various kinds of calmodulin-interacting agents have been proved to suppress tumor promotion in vivo. In this study, we further demonstrate that flunarizine and dehydroepiandrosterone, which are proved to interact with calmodulin, showed antitumor-promoting activity in two-stage carcinogenesis of mouse skin. These results indicate that Ca2+/-calmodulin system, may play an important role in tumor promotion. PMID- 2977965 TI - [Cryotherapy of metastatic B16 melanoma. Consequences of the immunogenic potential]. AB - A significant inhibition of B16 melanoma growth and prolonged survival of mice, immunized with cryolysat at 10 M omega of B16 melanoma cells, are obtained in an immunoprophylactic assays. These results show that the freezing maintains tumor rejection activity of tumor antigen. Immunization with frozen cells or their crude butanol extract induce only an inhibition of tumor growth without a prolongation of survival. PMID- 2977966 TI - [Methods of quick-freezing and cryosubstitution of biological tissues in analytical ion microscopy]. AB - The localization and relative concentration of diffusible ions (Li+, K+) or bound but easily displaceable ions (Ca2+) have been studied in myocardium prepared by rapid-freeze fixation and compared with results from chemically prepared samples. Contrary to chemical fixation which gives diffused potassium imaging, cryofixation enables elemental precise mapping in cellular compartment. Bound displazable external calcium, extracted by chemical fixation, is visualized after rapid-freeze fixation. Its localization is distinguished by a bright cellular outline, which corresponds to the glycocalix. Intracellular calcium is less emissive and has the same distribution and concentration whatever the sample preparation. Very movable and present at low concentration in tissues, lithium is localized after rapid-freeze fixation. Those qualitative and quantitative results show the adequation of rapid freeze fixation and freeze substitution to the study of diffusible ions by means of analytical ion microscopy. PMID- 2977967 TI - [Combined administration of sodium chloro-2-propionate and insulin on the secretion of glucagon by the pancreas in the diabetic rat]. AB - This work was designed to study the effects of sodium 2-chloropropionate (2CP) alone or combined with insulin, in vitro, on glucagon secretion from pancreas isolated from rats, made diabetic by streptozotocin (66 mg/kg i.p.). The pancreata were perfused with a physiological solution containing 2.8 mM glucose (0.5 g/l) and glucagon secretion was stimulated by an arginine infusion (5 mM) for 30 min. When 2CP (1 mM) and/or insulin (4 IU/l) were applied, they were infused from the start of the organ perfusion. In the presence of glucose alone, a marked decrease in glucagon output was observed in diabetic rat pancreas. The arginine perfusion induced a biphasic glucagon secretion both in normal and diabetic rat pancreas; this response was however clearly reduced in diabetic rat pancreas. In diabetic rat pancreas, the infusion of either 2CP or insulin had no effect on glucagon output in presence of glucose alone, nor did it modify the response to arginine. In contrast, the combined infusion of insulin and 2CP induced different effects depending on the conditions: whereas in presence of glucose alone it restored a glucagon output close to that recorded in normal rat pancreas, it did not modify the response to arginine. PMID- 2977969 TI - [Characterization of surface and intracellular glycoconjugates in larval forms of Echinococcus multilocularis using lectins]. AB - Carbohydrate moieties have been detected with nine fluorescent lectins in Echinococcus multilocularis cysts, developed in liver and lung of infested jirds. Tegument and glycocalyx, and two types of medullary cells were found to selectively and strongly bind lectins which are specifically adsorbed by N acetyl glucosamine, N acetyl galactosamine, galactose or mannose. The significance of these features in the host-parasite interaction are discussed. PMID- 2977968 TI - [Inter-organ differences of the cytometric DNA content in mice: relation of the staining method]. AB - With one step DNA staining methods including cell membrane lysis and RNase treatment, we regularly observed a higher fluorescence emission in liver nuclei compared to bone marrow nuclei in C57BL/6 mice. Therefore this study was conducted in order to emphasize such a phenomenon in other organs and to assess if higher fluorescence emission was related to higher DNA content or staining procedure failure. Liver, bone marrow and testis were removed from Swiss, BDF and C57BL/6 mice. The following samples were prepared: 1) liver cells with TRBC (TRBC = Trout Red Blood Cells = internal standards), 2) bone marrow cells with TRBC, 3) testis cells with TRBC and 4) mixtures of liver, bone marrow and testis cells. The staining procedures were: A) one step pH 10 procedure described by Vindelov (Virchows Arch. B. Cell Path., 1977, 24, 227-242), B) same procedure with twice RNase concentration, C) first method with twice NP 40 concentration and D) three steps procedure including Trypsin and Spermine treatment (Vindelov et al., Cytometry, 1983, 3, 323-327). In protocols A, B and C, "Diploid cells/TRBC" ratio differed significantly between liver, bone marrow and testis nuclei. Moreover, 3 distinct populations of diploid cells were present in samples 4. In protocol D, "Diploid cells/TRBC" ratio were identical between liver, bone marrow and testis nuclei. In samples 4, only 1 population of diploid cells has been observed. This study shows that DNA stabilization by polyamine and protein degradation by protease could act on Propidium Iodide fixation and/or fluorescence emission, with significant differences according to the origin of the cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2977970 TI - [Effects induced hypothermia on hepatic portal circulation]. AB - Studies of the liver circulation in dogs during hypothermia, showed that portal blood flow is reduced with no significant changes of blood pressure and with important vascular resistance. These effects are probably due to the contraction of pre-capillaries sphincters. PMID- 2977972 TI - A survey of children attending a rehabilitation centre in Harare. Implications for rehabilitation. PMID- 2977971 TI - [Effect of induced hypothermia on the systemic circulation]. AB - Studies of the systemic circulation in dogs (n = 5) during hypothermia showed that cardiac output, mean arterial pressure, total peripheral resistance, pulse rate, work L. V. is reduced and the stroke volume is increased. The authors think that these effects are probably due to metabolic alterations during hypothermia. PMID- 2977974 TI - [Interstitial photodynamic therapy of oral squamous cancers]. AB - Photodynamic therapy (PDT) was studied in the treatment of a variety of solid malignant tumors. Ten patients with oral squamous cancers treated by PDT are reported: The laser beam was introduced into the malignant and metastatic lymphatic tissues through inserted optic fibers. The results showed that all the lesions gave complete response to PDT except the bony involvement. It was noted that the lymphatic metastatic foci showed a higher photosensitivity, and the covering skin, connective tissue and the nearby organs were not obviously injured. Combination of PDT with chemotherapy, radiotherapy and surgery are also discussed. PMID- 2977973 TI - [Adrenal cortical carcinoma (ACC)--report of 10 cases]. AB - Ten patients with adrenal cortical carcinoma were treated from 1966 to 1986. There were 7 males and 3 females. The typical clinical manifestations, marked increase of 17-ketosteroid, 17-hydroxycorticoids and DHA, and negative dexamethasone suppression test were essential for the diagnosis. Of the ten patients, eight had secretive function and their 17-ketosteroid and 17 hydroxycorticoids varied from 36.8-93 mg% and 32.5-150 mg%, respectively. DHA was measured in 5 cases with the result of 6.95-44mg%. Those without secretive functions or obvious endocrine disturbances were usually misdiagnosed as kidney tumor, splenomagaly, liver tumor or pancreatic mass. Wood had summarized that nonsecretive ACC patients commonly had fever, pain, exhaustion syndrome (emaciation, fatigue, perspiration, anorexia), mass and distant metastasis. Adrenal scan, IVU, abdominal aortic arteriography, retroperitoneal pneumography and CT were helpful in localization. The differential diagnosis between ACC and adenoma by pathology was difficult. It is generally agreed that if the mass is larger than 100 grams, capsulated, having blood or lymphatic vessel invasion, hemorrhage, necrosis and calcification or even distant metastasis, malignant tumor should be considered. Surgical removal of the tumor is the only effective treatment. For advanced or recurrent lesions, selective adrenal artery thrombosis could be used. One of the ten patients was thus treated by this facilitated subsequent surgery. Postoperative chemotherapy, such as O.P-DDD, might be used in some cases. PMID- 2977975 TI - [Nature of cells with helper function for proliferation and differentiation of hemopoietic stem cells in the erythroid direction]. PMID- 2977976 TI - Changes in the plasma and urine alpha human atrial natriuretic peptide (alpha hANP) concentration in patients with thyroid disorders. AB - In order to assess the possible involvement of thyroid hormone in alpha human atrial natriuretic peptide (alpha hANP), we investigated the plasma and urine ANP concentration in patients with primary hyperthyroidism and hypothyroidism. Plasma and urine were extracted through Sep-Pak C18 cartridges and the urine ANP concentration was corrected by urine creatinine (cre. mg/dl) and expressed as fmol/mg.cre.. The plasma ANP concentration in patients with untreated hyperthyroidism (32.3 +/- 7.0 fmol/ml; n = 22) was higher than in normal subjects (p less than 0.01 vs control; 6.2 +/- 0.7 fmol/ml). After restoration to euthyroidism, the plasma ANP concentration (patients with treated hyperthyroidism) fell to normal (8.9 +/- 1.9 fmol/ml). The plasma ANP concentration in patients with untreated hypothyroidism (14.1 +/- 3.0 fmol/ml; n = 7) was higher than normal, but in two of them there was mild renal dysfunction and an incomplete right blundle branch block in the electrocardiogram. It was possible that these factors contributed to the observed increase in plasma ANP. However, a significant positive correlation was found between plasma ANP and free thyroxine (n = 40, r = 0.449; p less than 0.01) and free triiodothyronine (n = 40, r = 0.546; p less than 0.01). The urine ANP concentration in patients with untreated hyperthyroidism was markedly higher than in normal subjects (p less than 0.01), but in untreated hypothyroidism not significantly different from normal. PMID- 2977977 TI - Pathogenesis of extracellular fluid abnormalities of hypothalamic hypodipsia hypernatremia syndrome. AB - A 26-year-old man with hypothalamic hypodipsia-hypernatremia syndrome is reported, who presented with adipsia, hypernatremia, and impaired osmolality mediated arginine vassopressin (AVP) secretion. A chorionic gonadotropin secreting tumor was detected in the anterior hypothalamus and treated with external irradiation. After the treatment, hypernatremia persisted and was not corrected by fluid loading, osmolality-mediated AVP secretion remained impaired. Despite the absence of signs of hydropenia, hypovolemia was suggested by low blood pressure and elevated plasma indices of the renin-angiotensin system, and supported by blood volume determination. The plasma aldosterone concentrations were inappropriately low for the renin-angiotensin status. The plasma atrial natriuretic polypeptide (ANP) level was normal in spite of hypovolemia and increased more than double after fluid loading. Hypernatremia, primarily caused by hypodipsia and impaired osmolality-mediated AVP secretion, secondarily sustained ANP secretion and suppressed aldosterone release, which conceivably contributed to the development and perpetuation of hypovolemia in this patient. PMID- 2977978 TI - Effect of atrial natriuretic factor (ANF) on the secretory activity of zona glomerulosa in sodium-restricted rats. AB - Prolonged (12-day) sodium deprivation strikingly raised both basal plasma aldosterone concentration (PAC) (114%) and plasma renin activity (PRA) (200%), and lowered ANF blood level (-30%). Acute ANF bolus administration produced a dose-dependent decrease in PAC in both normally-fed and sodium-restricted rats. The maximum effect (-30/-37%) was observed with a dose of 20 micrograms.kg-1. The interruption of the renin-angiotensin system (RAS), obtained by a 7-day infusion of captopril and maintenance doses of angiotensin II, did not cause significant changes in PAC in animals kept on a normal diet, while it did induce a significant lowering of PAC in sodium-restricted rats (-25%). This treatment evoked in both groups of animals a notable reduction of PRA (-61/-89%). A 7-day infusion with ANF (at a rate of 20 micrograms.kg-1.h-1) notably lowered PAC ( 32%) in normally-fed rats, independently of the RAS status. The same occurred in sodium-deprived rats, but the effect was more intense in animals with intact RAS (-41% vs -24%). Prolonged ANF infusion significantly reduced PRA (-48%) only in sodium-restricted rats with intact RAS. These findings suggest that (i) the long term inhibitory effect of ANF on aldosterone secretion is due to both the block of renin release and a direct action on the zona glomerulosa; and (ii) the mechanism underlying the adrenoglomerulotrophic effect of sodium restriction involves not only the activation of RAS, but also the suppression of ANF release. PMID- 2977979 TI - [Medico-genetic studies of the Kostroma oblast population. X. Load of hereditary diseases in the population of Kostroma]. AB - Medical-genetic study of the population of Kostroma (the total size of the population analysed approx. 250,000) was carried on. The load of hereditary diseases in the population (per 1000) was 0.75 for autosomal dominant, 0.49 for autosomal recessive and 0.17 for X-linked recessive disorders. Significant differences in the prevalence of autosomal recessive hereditary disorders between rural populations and the population of Kostroma were observed. The dependence of the load of autosomal recessive pathology on random inbreeding was shown for the whole Kostroma province. PMID- 2977980 TI - [Cytogenetic effect of thaliblastine in a culture of human peripheral blood lymphocytes]. AB - The mutagenicity of thaliblastine (Bulgarian potential antitumor drug) was investigated in vitro in lymphocytes from healthy donors, and in vivo in lymphocytes of oncological patients after thaliblastine administration. No increase in the rate of chromosome aberrations was noted with increasing thaliblastine concentrations in vitro and in the course of therapy in vivo. Some polyploid metaphases were found in the lymphocytes of the patients treated with thaliblastine, as a result of the statmokinetic effect of the drug. Thaliblastine exerts extraordinarily slight mutagenic effect, as compared with other cytostatics. PMID- 2977981 TI - Effect of a single dose of mesoglycan on the human fibrinolytic system, and the profibrinolytic action of nine daily doses. AB - The profibrinolytic activity of orally administered Mesoglycan was evaluated in 18 patients affected by impaired plasma fibrinolytic activity. Mesoglycan was administered by a single oral dose of 24, 48 or 72 mg on 1 day, and by repeated doses of 48 mg twice a day for 9 consecutive days. After the single administration all the fibrinolytic parameters were significantly and positively influenced with an order of magnitude and a duration of effects proportional to the dose employed. After the repeated administration, a constant and reproducible activation of the fibrinolytic system was observed without any interference with haemocoagulative parameters. These results confirm that Mesoglycan is endowed with a relevant profibrinolytic activity in man after oral administration. The pharmacological activity of Mesoglycan could possibly involve the liberation of a certain amount of plasminogen tissue activator. PMID- 2977983 TI - Prevalence of respiratory symptoms and allergic sensitivity in multiply handicapped children. PMID- 2977982 TI - Cellular immunity and suppressor T cell function in asthmatic children on prolonged theophylline therapy. AB - Measurements of cellular immunity including the percentage of T-cell mitogenic response of lymphocytes to phytohemagglutinin (PHA) and concanavalin A (Con A), as well as Con A-induced suppressor function, were determined in 20 asthmatic children (mean age 12.4 +/- 0.6 years) before and after 6 months of theophylline therapy. Only 9 of the 20 children patients that commenced the study completed the entire 6 months of therapy. A group of 37 healthy children (mean age 12.6 +/- 0.3 years) served as control. During the 6-month period of therapy, theophylline levels were maintained between 10 and 20 micrograms%. No significant difference in any of the parameters examined were found between the initial 20 asthmatic patients and the control group. Furthermore, in the 9 patients that were evaluated before and after 6 months of theophylline therapy no significant change in lymphocyte function including suppressor function was detected. We conclude that the beneficial effect of long-term theophylline therapy in asthmatic children is probably not related to an immuno-modulatory effect. PMID- 2977984 TI - [Recent trend in the research of hypertension in Japan. Problems on the long-term treatment of hypertension--withdrawal of antihypertensive drugs and regression of cardiac hypertrophy]. PMID- 2977985 TI - [Effects of hexavalent chromium at non-lethal concentrations on the enzymology of the intestine of Salmo gairdneri and Dicentrarchus labrax (Pisces)]. AB - Effects of non lethal concentrations of hexavalent chromium on intestinal enzymology of Salmo gairdneri and Dicentrarchus labrax (Pisces). The effects of an exposure to potassium dichromate on intestinal enzyme activities (Alkaline phosphatase, maltase, leucine amino peptidase and ATPases) have been studied on a fresh water fish (Salmo gairdneri) and a salt water fish (Dicentrarchus labrax). Fish were exposed at seasonal temperatures (13 or 21 degrees C) to toxic concentrations equal to 1/10 of the 24 h-LC 50 (i.e. 18 mg/l Cr for trout and 5 mg/l Cr for bass) during respectively 13 and 21 days. Intoxicated trout stopped feeding and showed a decrease in their intestinal weight at the end of the experiments. A decrease of brush border membrane activities (Alkaline phosphatase, maltase and leucine amino peptidase) were also observed. These alterations have been interpreted as the consequence of the chromium induces fasting. Intoxicated bass showed no alterations of their feeding habits. Two specific effects of chromium on enzyme activities have been found: a severe decrease of the alkaline phosphatase activity and an increase of the Na/K ATPase activity. These enzyme activities could be useful indicators of chromium intoxication in marine fish. PMID- 2977986 TI - A specific ELISA using purified opsin, for studying autoimmunity in retinal diseases. AB - A highly sensitive enzyme-linked immunosorbent assay (ELISA) was developed to measure nanogram quantities of rhodopsin or its apoprotein, opsin, in bovine retinal rod outer segment (ROS) preparations. Anti-opsin anti-sera could detect as little as 4 ng of purified opsin or of opsin in ROS preparations. The purified opsin was prepared by quantitative elution from a preparative polyacrylamide gel, and showed higher immunoreactivity with anti-opsin than did ROS when the same amount (per weight) of protein was allowed to bind in the wells of the ELISA plates. The effect of the ionic detergent SDS (sodium dodecyl sulphate) on the immunoreactivity and antigen binding to the ELISA wells was studied. Concentrations of 0.1% SDS and above reduced the apparent binding of opsin with anti-opsin when examined by ELISA. This may have been because the negatively charged SDS reduced the efficiency of the antigen coating process, or because changes in the epitopes' conformations made them less recognisable by the corresponding antibodies. A similar ELISA system using a specific anti-S-antigen anti-serum allowed the detection of even very small amounts (nanograms) of S antigen in ROS preparations. The presence of S-antigen in ROS preparations was confirmed by immunoblotting. Thus purified opsin is preferable to ROS for ELISA tests of autoimmunity to rhodopsin in retinal diseases. These sensitive ELISA techniques could be used to examine the presence of minute amounts of rhodopsin, opsin or S-antigen in different retinal preparations. PMID- 2977987 TI - Spontaneous improvement of juvenile rheumatoid arthritis after T lymphocytosis with suppressor phenotype and function. AB - Spontaneous improvement of active juvenile rheumatoid arthritis (JRA) occurred after T lymphocytosis in an 8-year-old boy. He had prominent lymphocytosis, the count reaching 59,000/mm3, followed by spontaneous disappearance of fever, arthralgia, lymphadenopathy, hepatomegaly, and C-reactive protein. The serum immunoglobulin levels were gradually decreased. The surface marker analysis, using two color flow cytometry, showed that the lymphocytes were activated suppressor T lymphocytes, expressing CD3, CD8, HLA-DR, and CD8 plus CD11. When studied in vitro with pokeweed mitogen stimulation, the T lymphocytes significantly suppressed the immunoglobulin production by autologous B lymphocytes as compared with the T lymphocytes at remission (p less than 0.01). Based on the widely believed notion that depression of suppressor T lymphocyte functions is one of the important mechanisms underlying systemic JRA, the activated T lymphocytosis with the suppressor phenotype and suppressive function on the immunoglobulin production may have been related to the improvement of active JRA in the patient. PMID- 2977988 TI - Left ventricular hypertrophy as a possible indicator of systemic embolization in nonrheumatic atrial fibrillation--a clinical and echocardiographic study. PMID- 2977989 TI - Modulation of plasma glucose levels by thyrotropin-releasing hormone administered intracerebroventricularly in the rat. AB - Thyrotropin-releasing hormone (TRH), but not histidyl-proline diketopiperazine (cyclo[His-Pro]), induced transient hyperglycemia associated with hyperglucagonemia and marked hyperinsulinemia when placed intracerebroventricularly (i.c.v.) in anesthetized rats. This TRH-induced hyperglycemia was prevented by acute adrenalectomy. However, adrenalectomy did not prevent TRH-induced hyperinsulinemia or hyperglucagonemia. In streptozotocin induced diabetic rats, i.c.v. administration of TRH caused progressive and pronounced hyperglycemia. i.c.v. TRH-induced hyperinsulinemia was abolished by vagotomy and by systemic administration of hexamethonium or atropine. These results suggest that TRH induces hyperglycemia mediated by stimulation of the sympathetico-adrenal system and hyperinsulinemia by stimulation of the vagus nerve, and that the rapid decline of plasma glucose levels following transient hyperglycemia is due to hyperinsulinemia. PMID- 2977990 TI - [Studies on the role of heart innervation and other non-osmotic factors in regulating water and sodium metabolism]. PMID- 2977991 TI - PGO waves and insomnia in PCPA-treated rats. AB - The serotonin-depleting drug, parachlorophenylalanine (PCPA), in a dosage of 300 mg/kg, was administered to rats in an effort to test the hypothesis that altered distribution of PGO waves following drug treatment may be responsible for the sleep disruption and consequent sleep loss that accompany decreased serotonin levels. Consistent with the hypothesis, we found that the greater the proportion of PGO waves that precede spontaneous arousals, the greater the reduction in slow wave sleep. However, inconsistent with the hypothesis, we found that the decrease in sleep did not result from an increase in the number of arousals. Further, though an increase in the proportion of waking waves always accompanied a rise in wake time, the two variables were negatively correlated. These data do not support a PGO wave/arousal hypothesis to account for the decrease in sleep following PCPA treatment in the rat. Rather, the findings tend to implicate an alteration in the mechanisms of arousal linked to serotonin depletion. PMID- 2977992 TI - A comparison of feeding and locomotion responses to serotonin agonists in three rat strains. AB - The hyperphagic effects of two selective 5-HT1A agonists (8-OHDPAT and buspirone) in a free feeding paradigm and the locomotor suppressant effect of the serotonin agonist, m-chlorophenylpiperazine (m-CPP) were compared in three different rat strains: Wistar, Sprague-Dawley (SD), and Fawn-Hooded (FH) rats. Administration of various doses of 8-OHDPAT and buspirone produced significant increases in two hour food intake only in Wistar and SD strains and not in the FH strain. Similarly, various doses of m-CPP produced significant decreases in locomotor activity only in Wistar and SD strains and not in the FH strain. Isolated FH animals gained significantly less body weight relative to both Wistar and SD animals. These findings demonstrate attenuated feeding and behavioral responses to serotonergic agonists in the FH strain relative to both Wistar and SD strains. PMID- 2977993 TI - [Scavenging of active oxygen by selenium]. PMID- 2977994 TI - [Atriopeptin--a searched-for natriuretic hormone]. PMID- 2977995 TI - Personnel and patient doses during percutaneous transluminal angioplasty (PTA). PMID- 2977996 TI - [Atrial natriuretic factor]. PMID- 2977997 TI - [The inferior epigastric flap: experimental study]. PMID- 2977998 TI - [Endocarditis prophylaxis in dental interventions]. PMID- 2977999 TI - [Effect of Rhopalurus lauticauda venom on the isolated rectus abdominis muscle of Bufo marinus]. PMID- 2978000 TI - Endoscopy for contraception. PMID- 2978001 TI - Investigation of surface roughness of composite fillings. PMID- 2978002 TI - [Regression of infundibular hypertrophy of the right ventricle after pulmonary valvoplasty]. PMID- 2978003 TI - [Atrial natriuretic factor]. PMID- 2978004 TI - [Report on the meeting of the International Society and Federation of Cardiology and World Health Organization: task-force on coronary angioplasty]. PMID- 2978006 TI - [Synthesis of visual rhodopsin in a cell-free translation system. I. Influence of the structure of the synthetic bovine visual opsin mRNA on its translational efficiency]. AB - Influence of structural changes in nontranslated regions and translation initiation site of the in vitro synthesized bovine opsin mRNA on its translational efficiency in the wheat germ cell-free system has been studied. It is shown that level of the opsin synthesis up to 30 micrograms per 1 ml of translational mixture can be attained by optimizing structure of 5'-nontranslated region. PMID- 2978007 TI - [Oral characteristics of children with cerebral infantile paralysis]. PMID- 2978008 TI - [Tropomyosin from smooth and skeletal muscles initiates various conformational changes in skeletal F-actin]. AB - Changes in F-actin conformation in myosin-free single ghost fibers of rabbit skeletal muscle induced by the binding of skeletal and gizzard tropomyosin to F actin were studied by measuring intrinsic tryptophan-polarized fluorescence of F actin. It was found that skeletal and gizzard tropomyosin binding to F-actin initiate different conformational changes in actin filaments. Skeletal tropomyosin inhibits, while gizzard tropomyosin activates the Mg2+-ATPase activity of skeletal actomyosin. It is supposed that in muscle fibers tropomyosin modulates the ATPase activity of actomyosin via conformational changes in F actin. PMID- 2978009 TI - The internal dynamics of gene 32 protein-DNA complexes studied by quasi-elastic light scattering. AB - The hydrodynamic properties of large homodisperse single stranded DNAs complexed with the helix destabilizing protein of phage T4, the product of gene 32 (GP32), have been measured. The results suggest a size of the binding site between 8 and 10 nucleotides/GP32 molecule, in reasonable agreement with earlier work on a complex between GP32 and single stranded 145 base DNA. From static light scattering experiments it is concluded that the persistence length of these complexes is about 30 nm, distinctly smaller than the generally accepted value for double stranded DNA. The quasi-elastic light scattering properties of the DNA GP32 complexes were determined. The variation of the apparent translation diffusion coefficient Dapp with the scattering vector q was analyzed using the discrete ISMF and Rouse-Zimm models [S.C. Lin et al., Biopolymers 17 (1978) 425]. The model parameters that followed from the fit of Dapp versus q2 and from an extensive global analysis of the actually measured autocorrelation functions agreed with the notion that these DNA-protein complexes are indeed rather flexible. The continuous Soda model [K. Soda, Macromolecules 17 (1984) 2365] could successfully explain the variation of Dapp versus q2, assuming a persistence length of 30 nm and a base-base distance in the complex of 0.44 nm. PMID- 2978010 TI - [Osteogenic osteosarcoma, a model of a disease that is curable using a multidisciplinary treatment approach]. PMID- 2978011 TI - [Endogenous opiate modulators of insulin secretion in the obese]. AB - The effects of endogenous opiates on insulin response to oral glucose load were studied in obese subjects and in lean healthy volunteers. None of these having a family diabetes. After 3 days on an 1,800 cal./m2, 40% carbohydrate diet all subjects underwent two standard 75 g oral glucose tolerance tests (OGTT), one of which was accompanied by an i. v. administration of 10 mg of, an antagonist of opiates, the naloxone. In one group of obese impaired oral glucose tolerance test occurred. All obese, but not the lean healthy volunteers, showed: 1) increased basal plasma insulin levels, 2) higher insulin response to OGTT, 3) a decrease in insulin response to OGTT after naloxone administration, with significant differences at 60 min (p less than 0.01) and 90 min (p less than 0.025). In none of the subjects significant differences were observed in blood glucose levels after OGTT plus naloxone administration. These data suggest that increased endogenous opiates may affect insulin response to glucose in obese with impaired or normal oral glucose tolerance test. At present there seems to be no satisfactory explanation for unchanged blood glucose levels during OGTT with and without naloxone despite a decrease in insulin secretion in the obese patients. PMID- 2978012 TI - [Toxicology of cupric sulfate in honeybees. I. New algebraic parameters of lethality as alternatives to the LD50]. AB - A full kinetic study, over the whole life span of individuals, has been performed on the lethality of honeybees receiving by chronic administration: 0; 0.25; 0.5; 1; 2 g.l-1 of cupric sulfate in sucrose syrups. Three algebraic parameters, based on the Hill equation, allow a complete description of the curves: the time corresponding to the death of 50% of the individuals (LT50), the Hill coefficient (nt) and the slope of the tangent at the inflexion point (delta) corresponding to the maximum mortality rate. Studying the variations of the 3 parameters versus administered doses, leads to new relations allowing the calculation of the dose decreasing by half the 50% survival time of controls (dT50), and the doses increasing up to half the maximum, the value of nt(dnt50) and the value of delta (d delta 50). The experimental data gave: dT50 = 0.27 g.l-1; dnt50 = 0.51 g.l-1; d delta 50 = 0.93 g.l-1. These indexes, which represent new objective parameters for the characterization of lethality, should be substituted to the LD50 whose values vary between zero and infinite according to the time arbitrarily chosen for their calculation, and wear only a very restrictive significance. PMID- 2978013 TI - [Comparative effects of dihydropyridine inhibitors of calcium penetration on spontaneous contraction of the isolated heart atria in the guinea pig]. AB - All DHPs (nifedipine, nicardipine, nitrendipine) produced a concentration dependent depression of the isometric contraction and of the atrial rate of the isolated, spontaneously beating atria of the guinea-pig. The depressive actions of nifedipine and nitrendipine were completely antagonized by the addition of calcium, aminophylline and isoprenaline. Aminophylline partially, calcium almost completely and isoprenaline completely antagonized the depressive action of nicardipine on the isometric contraction. Only isoprenaline antagonized the effect of DHPs on the atrial rate of the isolated, spontaneously beating atria of the guinea-pig. It is possible that all these substances restore the contractibility of the atria by compensating the calcium balance, previously changed by DHPs, or by producing an increase in the intracellular cyclic AMP content (aminophylline and isoprenaline). PMID- 2978014 TI - [Effects of synthetic bioactive lipids on the activity of D-beta-hydroxybutyrate dehydrogenase, a membrane enzyme]. AB - The structural requirements of lecithins analogs for purified D-beta hydroxybutyrate dehydrogenase activation have been studied with chemically defined phospholipids. It appears that the trimethylamine group of choline can be changed by a pyridinium group. On the other hand, the decrease of density of the positive charges on the liposomes surface obtained by dilution of such bearing molecules with negative or non charged phospholipids increases the enzyme reactivation. Finally, the PAF acether, a lipid mediator, is able to reactivate the enzyme in similar conditions as these obtained with mitochondrial phosphatidylcholines. PMID- 2978015 TI - [Regression of mullerian ducts in the female chick embryo treated by norethindrone and gonadotropins]. AB - There is no correlation between the antigonadotropic activity of norethindrone and the regression of Mullerian ducts induced by this substance in the female chick embryo. PMID- 2978016 TI - [Quantitative approach of melanin in human cultured melanocytes]. AB - The purpose of the study was to establish an in vitro model for the quantification of the melanin produced in human melanocytes in culture. Melanin content in these cultured human melanocytes is determined by spectrofluorimetric assay. Fluorescence intensity is quantitatively related to the melanin content present in cultured melanocytes. PMID- 2978017 TI - [Effect of a diet containing calcium pantothenate on urinary vitamin excretion and on the liver and kidney total pantothenic acid level in rats]. AB - Four groups of five adult rats weighing 310 g received during 20 days a diet containing 0, 1.68, 16.8 or 168 mumol of pantothenic acid per kg of diet. The daily urinary vitamin excretion was, in nmol per day: 32 +/- 8, 32 +/- 4, 180 +/- 23 and 2,100 +/- 91, respectively (mean +/- SEM). Liver and kidney pantothenic acid content was the same in all groups, in nmol per g of fresh tissue: 300 +/- 36 and 190 +/- 6, respectively (mean +/- SEM, n = 20). PMID- 2978018 TI - Characterization of human platelet vasopressin receptor and the relation between vasopressin-induced platelet aggregation and vasopressin binding to platelets. AB - Immunoreactive AVP was found to be much higher in platelets than in platelet-free plasma (PFP) in normal subjects (12.8 +/- 6.3 versus 1.7 +/- 0.8 fmol/ml). AVP levels in PFP were appreciably elevated in parallel with the elevation of plasma osmolality induced by the acute osmotic stimulation, while the AVP levels in platelets did not change before and after the stimulation. Binding studies on intact platelets demonstrated specific binding sites for [3H]AVP. The specific binding was time, temperature and concentration-dependent, saturable and reversible, with the maximal binding capacity (Bmax) of 169.9 +/- 14.4 sites/platelet and affinity of 4.84 +/- 1.15 x 10(8)M-1. The affinity constants for unlabelled AVP, lysine vasopressin (LVP), oxytocin (OT) and dDAVP were 9.0, 8.5, 7.4 and 6.6, respectively, and the inhibition constant for d(CH2)5Tyr(Me)AVP (V1-antagonist) was 7.7. There was a highly significant correlation between the affinity constants of AVP analogues and their relative vasopressor activities in vivo, whereas no such correlation was found between the affinity constants and antidiuretic activities. AVP caused platelet aggregation with the maximal aggregation of 48.0 +/- 25.1% at 230 nM of AVP. A significant correlation was observed between the maximal percentage aggregation and Bmax of [3H]AVP to intact platelets. These results suggest that the platelet vasopressin receptor belongs to the V1 vascular subtype and mediates platelet aggregation. PMID- 2978019 TI - Isolation of U5 snRNP particles. AB - A method for the isolation of intact U5 small nuclear RNP particles from HeLa cells has been developed. The procedure includes nuclear extraction of the particles at moderate ionic strength, fractionation in agarose gels, electrophoretic transfer on DEAE cellulose paper and elution with ammonium chloride. The purified U5 snRNP particles contain U5 RNA and a set of eleven proteins. They retain their antigenicity as judged by their reaction with autoantibodies from patients with connective tissue diseases. PMID- 2978020 TI - U5 snRNP particles bind pre-messenger RNA in vitro. AB - The ability of purified U5 small nuclear RNP particles (U5 snRNPs) to bind in vitro pre-mRNA of the human beta-globin gene was investigated. The transcript which contained sequences, corresponding to the second intron and fragments of the second and third exons of the gene was bound to U5 snRNP particles in the presence of antibodies specific against U5 snRNPs. The in vitro splicing of pre mRNA was inhibited in the presence of autoantibodies against U5 snRNP particles. The results are discussed in terms of the possible role of U5 RNP particles in pre-mRNA splicing. PMID- 2978022 TI - Impairment of sexual behaviour in female rats with impaired glucose tolerance due to streptozotocin treatment of their maternal grandmothers. AB - A decreased receptive sexual behaviour was found in female F2-offspring of Streptozotocin-treated maternal grandmothers. These F2-offspring are characterized by decreased glucose tolerance and increased diabetes susceptibility, their metabolic situation being comparable to that of Type II prediabetes. The observed phenomena were associated with permanent hypoplasia of the hypothalamic ventromedial nucleus (HVN). It is hypothesized that there could be a causal relationship between these morphological changes in the HVN and impaired sexual behaviour in female prediabetic rats. PMID- 2978021 TI - The evaluation of androgen circulating levels following castration in adult male rats. AB - In the present study dehydroepiandrosterone (DHEA), androstenedione (A), testosterone (T) and dihydrotestosterone (DHT) plasma levels were determined in adult male rats five days, seven weeks and eleven weeks after orchiectomy and confronted, respectively, with rats 60 days old which were sacrificed 5 days, 7 weeks and 11 weeks after the sham-operation for orchiectomy. It was revealed that five days after castration A, T and DHT were decreased with respect to sham operated rats. Seven and eleven weeks after orchiectomy only T remained lower. In all three groups of castrated animals the A/T ratio resulted augmented whereas T/DHT ratio resulted lower with respect to the sham-operated animals. Five days after castration DHEA plasma concentration was positively correlated to A levels and both androgens resulted negatively correlated to T plasma levels. Seven weeks after bilateral orchiectomy an inverse correlation appeared between DHEA/A ratio and T. This emphasizes the role of DHEA and A in maintaining testosterone circulating levels. Seven and eleven weeks following castration the A/T ratio was negatively related to the T/DHT ratio, indicating that A contributes to DHT plasma levels. In the second part of our study the effect of sex steroid administration was evaluated 7 weeks after castration. A linear correlation between DHEA and T circulating levels was obtained following the administration of T while treatment with oestradiol caused a significant increase of the DHEA/A ratio. The castrated animals that received DHT presented lower T circulating levels while the A/T ratio was significantly increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978023 TI - [Psychiatric care and epidemiologic culture]. PMID- 2978024 TI - [Psychiatric epidemiology in the community: the importance of the definition of "case"]. PMID- 2978025 TI - [Evaluation of the operative style of psychiatric services: presentation and validation of a new questionnaire]. PMID- 2978026 TI - [Crisis and personal autonomy in patients at psychiatric facilities in the Tuscan region]. PMID- 2978027 TI - [Analysis of activity and use of psychiatric ambulatory services in Rome]. PMID- 2978028 TI - [First evaluation of the enactment of the Psychiatric Reform Act: function status and typologic models of new services in the light of the results of a national inquiry]. PMID- 2978029 TI - [Registration and monitoring of tumors in children (from 0 to 14 years of age) in the Umbria region]. PMID- 2978030 TI - [Assessment of causes of death in follow-up studies: comparison of procedures that can be used in Italy]. PMID- 2978031 TI - [Trends in psychiatric epidemiology]. PMID- 2978032 TI - [Manual of epidemiologic methodology]. PMID- 2978033 TI - Domiciliary care for the elderly patient. PMID- 2978034 TI - Long-term fluoride release from a composite restorative. PMID- 2978035 TI - Effect of different types of smear layers on dentin and enamel shear bond strengths. PMID- 2978036 TI - Aorto-Y-bifurcation graft: Dacron versus PTFE. Preliminary results of a randomized prospective study. AB - A randomized prospective study comparing PTFE-Y and Dacron-Y-grafts with regard to function rate, morphology, hemodynamic differences along the proximal anastomosis and body of the graft, complications and quality of material was performed. Between March 1983 and February 1987, 112 patients were admitted to the study. Randomization including the criteria of age, sex, indication to operation (chronic occlusive disease, aneurysm) run-off, diabetes, nicotine consumption and operative approach (transperitoneal, retroextraperitoneal) followed the methods of adaptive randomization developed by Pocock and Simon and was well balanced. Function rate and complications as well as morphological alterations showed no significant differences in either group. During a mean observation period of 24 months, Group I yielded a 97% and Group II a 95% function rate (Kaplan-Meier). In all patients but one in each group with limb graft occlusions (n = 5) function was regained by successful thrombectomy and profundaplasty. With respect to Doppler ultrasound differences in flow pattern, in four patients with enlarged proximal side-end anastomosis and adequate--not to large--incision is recommended performing the proximal anastomosis. PMID- 2978037 TI - The influence of quality on the long-term patency of arterial grafts: the surgeon's key role in improving results in medium and small caliber vessels. AB - The long-term patency of small and medium caliber grafts is directly affected by the surgeon's informed selection. In addition, when knitted synthetics are the appropriate choice, the methods used to preclot them for implantation will have a direct effect on their function. PMID- 2978039 TI - Nuclear medicine studies of aging--VI. Dual photon absorptiometry and bone scans in "at risk" women with back pain. AB - We studied 20 women who were at risk for osteoporosis (postmenopausal, or on dialysis, or on steroids), who presented with severe back pain. Patients underwent a dual photon (153Gd) densitometry study of the L2-L4 vertebrae, as well as an emission bone scan (99mTc-MDP). The dual photon method revealed abnormal results (20% or more below the mean expected value) in 13/20 patients (65%), and below the "fracture threshold" in 15/20 (75%). The emission bone scan delineated abnormal foci of uptake in 15/19 (79%). We conclude that women at risk for osteoporosis, who present with severe back pain, are likely to have a positive bone scan due to compression fracture or other derangements related to bone demineralization. PMID- 2978038 TI - New frontiers of vascular grafting. AB - Vascular grafting can be a very rewarding surgical technique: following surgery, most patients will maintain their autonomy and, in many cases, will be able to return to work. Should small diameter calibre substitutes be required, the autologous saphenous vein is the gold standard. If not available, the expanded PTFE is probably the best choice. For medium and large diameters, the polyesters (Dacron) are recommended provided they have a structure adapted to the needs of patients. However, progress in the last few years is not related to improvements in the grafts themselves. The only breakthrough has come with the success of graft seeding. This is the reason why other approaches have been proposed to restore or improve blood flow in atherosclerotic arteries: medication, sympathectomy, dilatation, laser endarterectomy, stent and spinal cord stimulation. All those approaches have more and more precise indications: they can supplement vascular grafting. PMID- 2978040 TI - Two-year study of passive-active immunization for prevention of hepatitis B infection in newborns. PMID- 2978041 TI - BMY-25801, an antiemetic agent free of D2-dopamine receptor antagonist properties. AB - BMY-25801, 4-amino-5-chloro-N-[2-(diethylamino)ethyl]2-(1-methyl-2-oxopropoxy ) benzamide, a new antiemetic agent free of D2-dopamine receptor antagonist properties, was effective against emesis induced by cytostatic agents (cisplatin, cyclophosphamide and doxorubicin) and total body radiation in the ferret. It also was effective against cisplatin-induced emesis in the dog; however, it was inactive against emesis caused by apomorphine and hydergine in the same species. In terms of activity profile, BMY-25801 could be differentiated both from metoclopramide and domperidone. Metoclopramide was nonselectively active against emesis induced by cytostatic agents, radiation and D2-dopamine receptor agonists, whereas domperidone was selectively effective against emesis induced by apomorphine and hydergine only. BMY-25801 failed to reveal any D2-dopamine receptor antagonist properties in several pharmacological tests (catalepsy, apomorphine stereotypy, serum prolactin, striatal dihydroxyphenylacetic acid and [3H]spiperone displacement) whereas metoclopramide was uniformly active in these tests. The activity profile of domperidone was compatible with its classification as a peripherally acting D2-dopamine receptor antagonist. BMY-25801 and metoclopramide antagonized serotonin-induced bradycardia (Bezold-Jarisch reflex) in the anesthetized rat, a response involving peripheral neuronal 5-HT3 receptors. Thus, BMY-25801 represents a novel antiemetic acting independently of D2-dopamine receptor mechanisms; however, its exact mode of action remains unknown. PMID- 2978042 TI - [Pathologic skin changes in workers at electric and thermoelectric power plants]. AB - Dermatological examination was performed and epidermal tests using a routine set of allergens and metals (Cr, Co, Ni, Al, Cu, Ag, Zn, Hg, Fe) on 112 workers of power plants and thermal-electric power stations working at the stands characterized by a heavy dustiness (electro-filters operation, ash removal, deslagging, carburizing) and at the stands where dustiness was not so heavy but instead exposure to machine oils and greases (retors' operators, electromechanics, assemblers and welders) was remarkable. It was found that occupational exposure to chemicals resulted in skin inflammation in 7.1% of the examined persons. Machine oils and greases induced skin inflammation in 2.7% and occupational acne in 5.3% of workers. It was also observed that chromium compounds were the primary allergen in workers exposed to dusts (13.4% of sensitized persons) and in workers exposed to industrial greases and oils (8.0% of sensitized persons). Allergy to cobalt compounds prevailed among persons exposed to smears and oils. Single positive results of epidermal tests with the use of copper and silver were obtained. Moreover, data concerning the microelements content in fly-ashes are presented. Information on the frequency of the incidence of occupational skin diseases, sickness absenteeism due to dermatoses and on personal safety equipment which should be used by the workers of power industry plants are provided. PMID- 2978043 TI - [Development and validation of a questionnaire in the study of spinal changes in a working population]. PMID- 2978044 TI - [Regional division by the level of endemicity of unilocular hydatid disease of the territory of the Azerbaijan SSR based on the results of a population examination]. PMID- 2978045 TI - [Seroepidemiologic characteristics of an echinococcosis focus in the Uzbek SSR]. PMID- 2978046 TI - [Seroepidemiologic characteristics of a trichinelliasis focus in Rostov Province]. PMID- 2978047 TI - [Comparative evaluation of 2 methods for demonstrating Borrelia in ticks--the vectors of Lyme disease]. PMID- 2978048 TI - [Structural features of the integration site of foreign DNA in the transgenic mouse genome]. AB - The structure of the transgenic mouse DNA region containing an integrated transgene (fragment of pBR322 sequence) was analysed. In one of the sequences flanking the transgene, short direct and inverted overlapping repeats were revealed at a distance of 60 bp from the integration site. In the same flanking sequence, there is an extended sequence (3.5 kbp) 0.3-1 kbp away from the transgene. It repeats 100-300 times in the mouse genome and is highly conservative (the homologs of the repeat have been revealed in other mammalian, bird, fish and insect genomes). This up-to-date unknown family of highly conserved dispersed repeats has been denoted by T1. We believe that both the revealed short inverted repeats capable of forming hairpins with loops and the T1 repeat are structures involved in the process of non-homologous insertion of foreign DNA into the region of the transgenic mouse genome. PMID- 2978049 TI - [Organization of simple sequences in the Drosophilia melanoga ter genome]. AB - Fragments of Drosophila melanogaster DNA that intensively hybridize with simple sequences poly[(dG-dT).(dC-dA)], poly[(dA).(dT)] and poly[(dG-dA).(dC-dT)] were cloned. The first two types of simple sequences are organized in these clones as separated stretches of moderate length, repeated many times within 12-15 kb. Each cluster contains only one type of the simple sequences and originates from a unique in the genome. In contrast, poly[(dG-dA).(dC-dT)] occurs in the genome as several isolated motifs. PMID- 2978050 TI - [The use of phage lambda promotors for expressing genes of secreted proteins in Bacillus subtilis cells]. AB - At was shown with the help of promoterless alpha-amylase and staphylokinase genes that lambda PR and lambda PL promoters could be used in Bacillus subtilis. Promoters strength was compared to promoter of alpha-amylase gene, this enabled to order the promoters in a row: PAA greater than lambda PR greater than lambda PL. The lambda PR promoter region was controlled by temperature in E. coli cells only, but not in B. subtilis, therefore, the active lambda C1857 gene product was not produced in B. subtilis cells. The lambda PR promoter is used by B. subtilis at a later growth stage than PAA and the lambda PL promoter at a still later stage than lambda PR. The data enables lambda PR to be considered as quite useful for Bacilli. PMID- 2978051 TI - Nigral dopamine autoreceptors are exclusively of the D2 type: quantitative autoradiography of [125I]iodosulpride and [125I]SCH 23982 in adjacent brain sections. AB - The effect of unilateral 6-hydroxydopamine lesions of the medial forebrain bundle on the specific binding of [125I]iodosulpride and [125I]SCH 23982 in the rat substantia nigra was determined by quantitative autoradiography of adjacent sections. The specific binding of [125I]iodosulpride was reduced by 40-70% on the lesioned side in the substantia nigra pars compacta, reticulata, lateralis and in the ventral tegmental area. In contrast, the specific [125I]SCH 23982 binding was unchanged in all subdivisions of the substantia nigra. The results indicate that dopamine autoreceptors are present in the substantia nigra and in the ventral tegmental area and that they are exclusively of the D2 type. PMID- 2978052 TI - Atrial natriuretic factor in sympathetic ganglia of the rat: dependence on cholinergic innervation. AB - Atrial natriuretic factor is detectable in the peripheral autonomic ganglia of the rat by radioimmunoassay and immunohistochemistry. In the present study, surgical and neurochemical methods were used to evaluate the source of this peptide in sympathetic ganglia. Decentralization of the ganglia and/or central administration of colchicine diminished the atrial natriuretic factor content in para- and prevertebral ganglia. Axotomy did not affect levels of ganglionic atrial natriuretic factor. A messenger ribonucleic acid species hybridizing with rat atrial natriuretic factor complementary deoxyribonucleic acid was not found within the total ribonucleic acid extracted from superior cervical ganglia. These results indicate a direct dependence of ganglionic atrial natriuretic factor on cholinergic innervation. PMID- 2978053 TI - [Contact allergy to ethylenediamine]. PMID- 2978054 TI - [A proposed new method of determining the properties of allergenic substances used in industry]. PMID- 2978055 TI - [Occupational allergy in dentists]. PMID- 2978056 TI - [Occupational allergy in nurses]. PMID- 2978057 TI - [AIDS: oral manifestations and prevention in dental practice]. PMID- 2978058 TI - [Luetic diseases]. PMID- 2978059 TI - [Management of emergencies of hepatic origin]. PMID- 2978060 TI - [Hematological aspects of the dental patient]. PMID- 2978061 TI - [Tuberculosis]. PMID- 2978062 TI - [Gonorrhea]. PMID- 2978063 TI - [Pulmonary diseases in dental practice]. PMID- 2978064 TI - Episodic barrel rotations induced by intrastriatal injection of quinolinic acid in rats. Inhibition by anticonvulsants. AB - Unilateral intrastriatal injection of quinolinic acid (2,3 pyridine dicarboxylate; QUIN) in the rat produces episodic barrel rotations and tonic clonic forepaw movements, lasting for several hours. We investigated whether intraperitoneal posttreatment with anticonvulsants could abolish this phenomenon when it is already fully developed, and whether their potency ratio was similar in models of epilepsy. All 8 tested antiepileptics, namely carbamazepine, clonazepam, diazepam, diphenylhydantoin, ethosuximide, flunarizine, phenobarbital and sodium valproate decreased this behaviour in a dose-dependent way. Six other drugs with anticonvulsant properties were also effective: DL-2-amino-7 phosphonoheptanoic acid, desipramine, etomidate, ketamine, meprobamate and sabeluzole. The ED50-values for halving the frequency of the episodes of barrel rotation correlated well with published ED50-values for inhibition of tonic hindpaw extension in the maximal metrazol seizure test (rs = .95, p less than 0.001) and with the ED50-values for halving the duration of the forepaw clonus in the rat-kindling model (rs = .93, p less than 0.001). This quinolinic acid test allows visualization of the onset of action of anticonvulsants, with each animal as its own control. In order to assess whether this test is also sensitive to drugs influencing the symptoms of Huntington's disease, the effect of the dopamine antagonists haloperidol and pimozide, the acetylcholinesterase inhibitor physostigmine and the anticholinergics atropine and dexetimide were investigated as well. The experiments suggested that the barrel rotations and clonic forepaw movements, only 3-6 hours after intrastriatal injection of QUIN respond to anticonvulsants, but are not specifically sensitive to drugs used in the symptomatic treatment of Huntington's disease. PMID- 2978065 TI - i.r. Beta-endorphin, corticosterone, cholesterol and triglyceride concentrations in rat plasma after stress, cingulotomy or both. AB - Plasma i.r. beta-endorphin, corticosterone, cholesterol and triglyceride concentrations were determined in male Sprague-Dawley rats after exposure to running or swimming stressors or after surgical ablation of the cingulum bundle. While cingulotomy alone altered only the plasma triglyceride concentrations, the combination of cingulotomy plus running stress significantly increased plasma i.r. beta-endorphin and triglyceride concentrations above those seen in animals receiving only a running stress. Triglyceride concentrations in cingulotomy plus swimming stress were significantly elevated above those in animals receiving a severe stress only. While the exposure to running and swimming increased plasma beta-endorphin significantly above control levels, plasma corticosterone was not affected by these stressors. Changes in plasma cholesterol and triglycerides were also differentially affected by cingulotomy or stress exposure. These results indicate that various stress hormones are affected differentially by exposure to various experimental procedures which are employed as stressors. PMID- 2978066 TI - [News in AIDS research--information for the dentist. Contradictory preventive strategy]. PMID- 2978067 TI - [Current views on the pathogenesis and treatment of aplastic anemia]. PMID- 2978068 TI - Rehabilitation of a nutritionally and dentally compromised patient. PMID- 2978069 TI - [Bonding of composites to dentin treated with Tubilicid]. PMID- 2978070 TI - [Indications for and results of anterior resection of the rectum]. PMID- 2978071 TI - [Late results following arthroplasty using a Moore prosthesis]. PMID- 2978072 TI - [The role of surgical intervention in the combined treatment of advanced ovarian neoplasms]. PMID- 2978073 TI - [Choledochoduodenostomy in benign hepatobiliopancreatic lesions]. PMID- 2978074 TI - [The Harrison-Debas operation in traumatic rupture of the duodenum]. PMID- 2978075 TI - [Paraneoplastic polyarthritis revealing gastric cancer]. PMID- 2978076 TI - [A case of xanthogranulomatous pyelonephritis]. PMID- 2978077 TI - [Xanthogranulomatous pyelonephritis. Comments apropos 145 clinical cases]. PMID- 2978078 TI - [Riedel's thyroiditis]. PMID- 2978079 TI - [Complications due to excessive intravascular length of a central venous catheter]. PMID- 2978080 TI - [Ocular tension in diabetics]. PMID- 2978082 TI - [The value of the ophthalmologic examination in detecting rhinosinusal lesions of tumor origin]. PMID- 2978081 TI - [Antigen S and ocular autoimmunity]. PMID- 2978083 TI - [The importance of studying HLA histocompatibility antigens in eye lesions]. PMID- 2978084 TI - [And eye diseases are disappearing?]. PMID- 2978085 TI - [Microscopic and quantitative aspects of the human cornea as a function of age]. PMID- 2978086 TI - [Treatment of cataract with Quinax]. PMID- 2978087 TI - [Fukala's operation in marked myopia]. PMID- 2978088 TI - [Problems arising in the treatment of monocular traumatic cataract. The inverted artificial crystalline lens]. PMID- 2978089 TI - [Scleroplasty in degenerative myopia. Postoperative results]. PMID- 2978090 TI - [Orbito-ocular traumatism]. PMID- 2978091 TI - [Exophthalmos due to ethmoidal and orbital mucocele]. PMID- 2978092 TI - [A guide to the ophthalmologic care of children with retinopathy of prematurity]. PMID- 2978093 TI - [The glass-ionomers in adhesive dentistry: a rational material. 2. Clinical use]. PMID- 2978094 TI - Immunopathology of human schistosomiasis mansoni. I. Immunomodulatory influences on T cell function. PMID- 2978095 TI - Skin circulation in the nipple after reduction mammaplasty by upper and lower glandular resections. AB - Avascular necrosis of the nipple is a serious complication of reduction mammaplasty with nipple transposition. A study was undertaken to measure the skin circulation in the nipple before, during and after this operation. In 14 patients undergoing a reduction mammaplasty according to the method of Strombeck, the skin circulation was measured in 25 breasts with laser doppler flowmetry (LDF) and fluorescein flowmetry (FF). LDF showed that the skin circulation increased after de-epithelialization to 204.4 +/- 31.0% of the preoperative value (100%) (mean +/ SEM, p less than 0.01). After the upper and lower glandular resection the circulation was reduced to 90.7 +/- 12.3% of the preoperative value. The division of the lateral pedicle did not affect the circulation. After the skin had been sutured, the circulation was 71.5 +/- 9.1% of the preoperative value (p less than 0.01). One to four days postoperatively the circulation was 100.3 +/- 13.2% of the preoperative value. At FF uniform fluorescence was observed in the nipple postoperatively in all patients but two, in whom avascular necrosis later developed. Our results thus show that the circulation in the nipple after reduction mammaplasty by the Strombeck method is adequate and that it is safe to divide the lateral dermal pedicle. PMID- 2978096 TI - [Fibromyalgia (fibrositis). A heterogenous pain syndrome]. PMID- 2978097 TI - [Histomorphologic changes in the lung tissue in death from overcooling]. PMID- 2978098 TI - A flexible genetic code, or why does selenocysteine have no unique codon? PMID- 2978099 TI - [Disodium cromoglycate in comparison with antihistaminics (loratadine) in seasonal allergic rhinitis]. PMID- 2978100 TI - [Reproducibility of epicutaneous test results]. AB - 857 non-dermatologic patients were patch-tested with the Finn-Chamber-technique. The sequent testings under identical conditions were made after 3-12 months. Readings were done after 24, 72, 96 hours and later. During the first test-series we found 44 reactions to be positive, 20 remained identical during repetition testings, 7 became negative and 36 positive reactions appeared. So reproducibility was 46% for the positive results of the first testing serie. PMID- 2978101 TI - [Modern methods of acne therapy]. AB - Today the treatment of acne vulgaris offers a broad spectrum of topic and systemic therapeutics. With these a optimal therapy in different acne-phases is practicable, and the possibility of scar formation is inhibited. The particular groups of substances are discussed regarding indication, action and contraindication. PMID- 2978102 TI - [Dermatologic problems of workers in clean rooms]. AB - Special dermatological problems of workers in clean-rooms. Ever since the development and production of microelectronic parts require so called working conditions of "clean-rooms", dermatologists are increasingly confronted with skin problems of persons working within this environment. The skin problems associated with excessive dryness are attributable to low humidity in the "clean-rooms" and as an additional factor to the lamellar air flow of filtered air. With people working in clean rooms--protective clothing notwithstanding--the incidence of skin flaking is high, and this can cause electronic faults. Even components of normal skin-care agents can be the cause of such electronic faults. Use of skin cosmetics, virtually free from inorganic substances, can lead to a considerable reduction in skin flaking and prevention of exsiccation. PMID- 2978103 TI - [Skin protection as an occupational dermatologic problem]. AB - Skin protection has a particular importance in occupational dermatology. The skin has to be protected from different occupational noxae for example physical like heat, cold, UV-radiation or friction or chemical like toxic or allergenic contactants. The protection measures are specified, in detail new formation of workrooms to prevent irritation of the skin, protective clothes and gloves, different noxa-adapted, skin protective preparations including different detergents and care of the skin. Their valence concerning prophylaxis and present occupational eccema is discussed. PMID- 2978104 TI - Laparoscopy for screening and early diagnosis of ovarian cancer. AB - Authors examined validity of laparoscopy for early diagnosis of ovarian cancer. They studied 144 cases of patients with benign neoplasia diagnosed by clinical examination and found ten cases of ovarian cancer by laparoscopy. They point out validity of laparoscopy indicated as a present day essential technique for early diagnosis of ovarian cancer. PMID- 2978105 TI - [Studies on platelets and coagulofibrinolytic system in ischemic heart disease]. PMID- 2978106 TI - [Hepatic porphyria and dentistry]. PMID- 2978107 TI - [Preparation of a patient with carcinoma of the mouth before radiation]. PMID- 2978108 TI - [Convulsive attacks and the dentist]. PMID- 2978109 TI - [The role of pedodontics in cleft treatment strategy]. PMID- 2978110 TI - [Principle infectious complications of heroin addition: their incidence in stomatology]. PMID- 2978111 TI - [Hereditary angioneurotic edema in oral surgery]. PMID- 2978112 TI - Bactericidal activity of M14659 enhanced in low-iron environments. AB - The bactericidal activity of M14659 against Escherichia coli in low-iron environments was investigated and compared with that of ceftriaxone and ceftazidime. The bactericidal activity of M14659 against E. coli in Mueller Hinton broth was enhanced 30- to 20,000-fold by addition of transferrin, which is an iron-binding protein, whereas the activity of ceftriaxone or ceftazidime was much less strongly affected. This enhancement by transferrin was completely inhibited by saturating the iron-binding capacity of transferrin with FeCl3. M14659 was taken up markedly into bacterial cells in the presence of transferrin, and its uptake was inhibited by the protonophore dinitrophenol, which inhibits active-transport systems coupled to an energized membrane such as the iron transport systems of E. coli. The bactericidal activity of M14659, which chelates Fe3+, was also enhanced in the presence of other iron-binding compounds such as lactoferrin and alpha,alpha'-dipyridyl or in iron-deficient Mueller-Hinton broth (Fe3+ concentration, less than 2 nM) supplemented with FeCl3 at 0.1 to 1.0 microM, but not in unsupplemented iron-deficient Mueller-Hinton broth. The E. coli used in this study was confirmed to derepress iron transport systems in the presence of transferrin, lactoferrin, and alpha,alpha'-dipyridyl and in the iron deficient Mueller-Hinton broth supplemented with FeCl3 at 0 to 1.0 microM. M14659 also showed an excellent antibacterial activity in vitro against other gram negative bacteria in the low-iron environments. These findings indicate that M14659 may be actively taken up with Fe3+ into bacterial cells, probably through the iron transport systems under conditions of low iron and, thus, kills bacteria effectively. PMID- 2978113 TI - The role of Lyt-2+ T cells in the regulation of autoimmunity in murine lupus. AB - Systemic lupus erythematosus (SLE) in humans and in mice is characterized by reduced suppressor T-cell activity. This observation suggests that selective loss or impaired function of suppressor T cells may contribute to the development of autoimmunity. To clarify the role of suppressor T cells in the pathogenesis of SLE, we used a rat MAb to selectively deplete Lyt-2+ ('suppressor/cytotoxic') T cells from lupus-prone NZB/NZW F1 (B/W) mice. Treatment consisted of weekly intraperitoneal injections of anti-Lyt-2 (2 mg/mouse) beginning at age 4 months, prior to the onset of overt clinical illness. Control mice received weekly injections of either non-immune rat IgG or saline. Despite sustained depletion of Lyt-2+ T cells, mice treated with anti-Lyt-2 were indistinguishable from control mice with respect to production of anti-DNA antibodies, development of renal disease, and mortality. These findings imply that Lyt-2+ T cells do not regulate autoimmunity in B/W mice. However, they do not exclude the possibility that Lyt 2+ T cells suppress autoimmunity in normal mice but are simply non-functional in B/W mice. Therefore, we also examined the consequences of depleting Lyt-2+ T cells from non-autoimmune C57BL/6 x NZW (B6/NZW) mice. Depletion of Lyt-2+ T cells from B6/NZW from age 4 to 10 months produced neither serologic nor clinical evidence of murine lupus. These observations suggest that suppressor T cell defects are not sufficient to cause murine lupus. PMID- 2978114 TI - T cells expressing gamma delta chain receptors in rheumatoid arthritis. AB - Whereas the majority of T cells use alpha and beta chains to form their T-cell receptor, a small minority of T cells, which do not express the CD4 or CD8 surface markers, use other chains termed gamma and delta to form their receptor. Flow cytometry was performed on cells isolated from the blood and synovial joints of patients with rheumatoid arthritis. Monoclonals which recognise the gamma and delta chains were used to compare the proportion of TCR gamma delta cells in these sites. Approximately half the patients had more TCR gamma delta in the joints than in their blood and one newly diagnosed patient had high numbers of TCR gamma delta cells in both blood and joints. In this preliminary study it is not possible to evaluate the role of these cells in the disease process, but it is of interest that in some RA patients there is an overabundance of both T cells that arise early in ontogeny (TCR gamma delta cells) and B cells that arise early in ontogeny, the CD5 B cell. PMID- 2978116 TI - "We want water, not gold". PMID- 2978115 TI - Defective induction of T-cell help and natural killing following anti-CD3 stimulation of autoimmune lymphocytes. AB - Anti-CD3 monoclonal antibody (MoAb) stimulates T cells in normal peripheral blood to proliferate and develop cytotoxic activity against NK-sensitive tumor cell lines. We now find that anti-CD3 MoAb also generates cytotoxic activity against a cell line (MEL-21) resistant to classical NK cell killing. After activation in vitro with anti-CD3 MoAb for 18 h, normal peripheral blood mononuclear cells (PBMNC) develop more HLA-DR-positive helper than suppressor T cells, manifest a functional helper effect as measured by increased IgG synthesis (P less than 0.01), as well as kill MEL-21 target cells. PBMNC from rheumatoid arthritis (RA) patients respond normally but mononuclear cells from rheumatoid arthritis synovial fluid (RASF) respond poorly. PBMNC from systemic lupus erythematosus (SLE) patients also respond poorly to anti-CD3 stimulation. Thus, the ability of anti-CD3 to stimulate IgG production and generate enhanced natural cytolytic activity are defective in both RASF and SLE lymphocytes. PMID- 2978117 TI - Cerebral degenerations producing dementia: importance of neuropathologic confirmation of clinical diagnoses. AB - Dementia is a major public health concern with our increasing elderly population and currently affects more than three million Americans at an annual cost of $50 billion. The marked overlap in symptomatology between Alzheimer's disease and other primary parenchymal degenerations makes antemortem diagnosis based on clinical assessment tentative at best, with error rates of 25% commonly reported. Accurate diagnosis is of vital importance in order to improve our understanding of these illnesses, evaluate potential therapies, and provide appropriate genetic counseling to family members. Direct neuropathologic examination at autopsy is currently the only reliable method for assuring accurate diagnosis, and should be undertaken in all demented patients. To illustrate the importance of these principles, we present three patients who were clinically diagnosed with Alzheimer's disease, and subsequently found to have other dementing illnesses by careful postmortem neuropathologic examination. PMID- 2978118 TI - Markedly increased platelet membrane fluidity in Down syndrome with a (14q, 21q) translocation. AB - Abnormal platelet membrane fluidity was found in a 50-year-old woman with Down syndrome who had been experiencing a gradual mental decline. The patient's karyotype showed a triplication of the long arm of chromosome 21. The authors hypothesize that the PMF-locus, which carries the allele for increased membrane fluidity, is located on the long arm of chromosome 21. Similarities between Down syndrome and Alzheimer's disease and the possible relationship between membrane fluidity and cognitive function are mentioned. PMID- 2978119 TI - Theoretical and experimental study of the time dependent flow of red blood cell suspension through narrow pores. AB - The Hemorheometer has been adapted to allow the recording of the flow rate during the filtration process. For newtonian fluids, the flow rate variation versus time through the pores is well approximated by Poiseuille's law. For dilute red blood cell suspensions, the same analysis can be applied by introducing the concept of "apparent filtration viscosity" which is higher than the usual viscosity measured by Couette viscometry. The apparent filtration viscosity parameter is related to the deformations undergone by red blood cells as they pass through the narrow pores. Apparent filtration viscosity can be used to obtain a precise determination of the erythrocyte deformability. Measurements performed, for a given blood sample, with pores of different diameters (5 microns, 8 microns and 12 microns) show that the error on the value of apparent filtration viscosity is less than 3%. As a result, the sensitivity of the filtration method allows to discriminate among normal blood samples. High concentrations of erythrocytes or leucocytes are found to modify the apparent filtration viscosity. These factors are apparent in the recorded filtration curves. Their effects on filtration measurements can be easily estimated. PMID- 2978120 TI - The effect of hematoporphyrin derivative and human erythrocyte ghost encapsulated hematoporphyrin derivative on a mouse myeloma cell line. AB - The effect of hematoporphyrin derivative (HPD) in combination with HeNe laser radiation on a mouse myeloma cell line, P3X63Ag8U1, has been examined. The effects of varying doses of radiation and photosensitizer were assessed using both dye exclusion and clonogenic assay systems. Development of photosensitivity by those cells in the presence of hematoporphyrin (HP) and HPD has been examined. In addition, with a view towards assessing erythrocyte encapsulation as a vehicle for an immunotargeting system for HPD, the effect of HPD encapsulated in such a manner, on the mouse myeloma cell line has been examined. From the results obtained from these in vitro experiments the authors believe that this may be a convenient means of packaging HPD for antibody-mediated delivery to tumour cells. PMID- 2978121 TI - Glomerular basement membrane heparan sulfate glycosaminoglycan in aminonucleoside of puromycin nephrosis. AB - Proteinuria in aminonucleoside of puromycin (PAN) nephrosis is due to an alteration of the glomerular charge- and size-selective barrier. Heparan sulfate seems to play a role in glomerular charge permeability to plasma proteins. We report a quantitative analysis and metabolism of glomerular basement membrane (GBM) heparan sulfate in PAN nephrosis. Studies were performed 5 and 10 days post PAN administration (15 mg/100 g) to Sprague-Dawley rats. GBM heparan sulfate glycosaminoglycan was measured using the dimethylmethylene blue test after chromatographic separation of the digested GBM. Sulfate-35 uptake by GBM and catabolism of GBM-sulfated compounds were studied using glomerular cultures. The heparan sulfate glycosaminoglycans concentration on day 5 (1.42 micrograms/mg GBM protein) was within the normal range (1.17 +/- 0.25), but the concentration on day 10 was below the limit of detection (0.57 micrograms/mg). The sulfate-35 incorporated in the GBM glycosaminoglycan on day 5 (158 cpm/micrograms glycosaminoglycan) was lower than the uptake of control GBM (270 cpm/micrograms glycosaminoglycan), and markedly lower than the uptake on day 10 (1,590 cpm/micrograms glycosaminoglycan). The catabolism of the GBM sulfated compounds on day 5 and 10 was not different than the one seen in controls. These data suggest that PAN administration is associated with a decrease in synthesis of GBM sulfated compounds on day 5 and a decrease in GBM heparan sulfate glycosaminoglycan on day 10. However, on day 10, an increased compensatory synthesis is observed. This may subsequently normalize the GBM heparan sulfate concentration and explain the resolution of the proteinuria. PMID- 2978122 TI - Changes in plasma concentrations of atrial natriuretic peptide during the first ten days and the second month of life. AB - Atrial natriuretic peptide (ANP) was assayed on the 1st, 3rd, 7th and 10th day of extrauterine life and the levels of the hormone were checked after the 2nd month in 12 infants. After an initially rapid increase, with a peak of the 3rd day (45.7 +/- 7 pg/ml the 1st day of extrauterine life and 175 +/- 5 pg/ml on the 3rd day), we can recognise a progressive decrease until the measurement made at 2 months (61.5 +/- 5 pg/ml). The reported data show the important role of ANP in adaption of life after birth. This can be considered as a compensatory response because it represents an attempt to maintain the balance of positive sodium which is essential for the growth of the newborn. PMID- 2978124 TI - The effect of atrial natriuretic factor on arginine-8-vasopressin and oxytocin levels in various brain regions and plasma. AB - The effect of intracerebroventricular (i.c.v.) treatment of rat atrial natriuretic factor III (ANF III; 0.5 microgram) was measured on the arginine-8 vasopressin (AVP) and oxytocin (OXT) contents of rat hypothalamic and limbic brain areas as well as those in the plasma. The hormone concentrations were determined by radioimmunoassay (RIA). The administration of ANF III in conscious euhydrated rats resulted in a significant reduction of both AVP and OXT contents in the hippocampus. Ether anesthesia interfered with the effect of ANF III, since in anesthetized rats ANF III reduced the levels of AVP and OXT in the septal regions, too. ANF III had no effect on the basal plasma AVP and OXT concentrations, however, the peptide inhibited the plasma AVP and OXT elevation induced by hyperosmosis (intraperitoneal injection of 2.5% NaCl). The results suggest that ANF III may be important in the control of the activity of both the peripheral (hypothalamo-neurohypophyseal) and the central (brain) AVP-ergic and OXT-ergic systems. PMID- 2978123 TI - Ovulation induction for in-vitro-fertilization and embryo transfer applying decapeptyl (DTRP-6 LH/RH) in combination with HMG or FSH. AB - 47 patients out of the IVF-program of the Department of Obstetrics and Gynecology University of Kiel, who demonstrated in previous stimulation cycles premature LH surges, were treated in two modalities with a down regulation applying the GnRN analogue decapeptyl (DTRP-6 LH/RH) and a concomitant HMG- or FSH-stimulation. The down-regulation was started after ovulation up to a negative LH/RH test followed by a concomitant gonadotropin stimulation in group 1. In group 2 a parallel treatment with decapeptyl and HMG or FSH was performed from day 2 of the cycle. 10 husbands of the punctured patients had pathological sperm. In both groups 6 patients were discarded from of the stimulation protocol as their oestradiol responses were not adequate, sperm contamination was detected late in one case, and in one patient a premature LH surge occurred once again. In 36 patients vaginal follicular punctures were performed. With respect to pregnancies group 1 revealed a much higher pregnancy rate than group 2. It seemed better to start the down-regulation with the GnRH-analogue decapeptyl in the luteal phase of the previous cycle. The treatment with decapeptyl should not only be applied in patients with previous LH surges but also in order to establish a synchronous follicular maturation in ovulating patients treated for in-vitro fertilization, or gamete-intra-Falloppian-tube-transfer. PMID- 2978125 TI - [From symptom to risk]. PMID- 2978126 TI - [ISTAT surveys on the health status of the Italian population and the use of health services: 1980 and 1983]. PMID- 2978128 TI - [Incidence of tumor of the female breast in the province of Florence from 1977 to 1982]. PMID- 2978127 TI - [Estimate of the hospital incidence of schizophrenia in Lombardy]. PMID- 2978129 TI - [Screening for colorectal neoplasms at a regional corporation]. PMID- 2978130 TI - [Description and evaluation of the information system of the departments of mental health in the Lazio Region]. PMID- 2978131 TI - [Evaluation of ophthalmologic pediatric screening at 3, 12, and 24 months of age]. PMID- 2978132 TI - [Unemployment and health: what Italian statistics show]. PMID- 2978133 TI - [Physical activity of school age children and prevention of cardiovascular diseases]. PMID- 2978134 TI - [Exposure to electromagnetic fields with extremely low frequency and occurrence of tumors in man: evaluation of epidemiologic studies]. PMID- 2978135 TI - [The nature of tumors and violins]. PMID- 2978136 TI - [Mortality of in-patients at public psychiatric hospitals in the Lazio Region: first results]. PMID- 2978137 TI - [The first ambulatory visits to psychiatric services in the Emilia-Romagna region. Evaluation of incidence and sociodemographic factors]. PMID- 2978138 TI - [Cultural attitudes in the comparison of alcoholic beverages: a survey on teachers of the local health unit 18 of the Veneto Region]. PMID- 2978139 TI - [A school health service at a local health unit in Veneto: evaluation and proposals]. PMID- 2978140 TI - [Use of a regional file of hospital discharges for a cohort study]. PMID- 2978141 TI - [Tumors of the lung attributable to occupational exposure]. PMID- 2978142 TI - [Use of administrative data for epidemiologic research. Consultation of the Fiscal Code Archive for ascertaining living existence in cohort studies]. PMID- 2978143 TI - [The proportion of tumors of the lung caused by occupational exposure: updating]. PMID- 2978144 TI - [Secondary prevention may also be true prevention]. PMID- 2978145 TI - [Methodologic debate on the design of the controlled case study]. PMID- 2978146 TI - [Occupation-exposure matrix for 16 carcinogenic or suspected carcinogenic substances for the respiratory system]. PMID- 2978147 TI - [Identification of distortion due to misclassification and selection errors in studies of analytic epidemiology. A bibliographic review]. PMID- 2978148 TI - [Recommendations for screening in oncology]. PMID- 2978149 TI - [A longitudinal system of mortality surveillance according to socio-economic characteristics, as shown by population censuses: description and documentation of the system]. PMID- 2978150 TI - [Mortality and occupational status in the Turinese longitudinal study]. PMID- 2978151 TI - [Years of potential life lost in Italy before 65 years of age]. PMID- 2978152 TI - [A case of general anesthesia for the dental patient with cat cry syndrome]. PMID- 2978153 TI - [Correlation between arterial pressure at rest and during effort and left ventricular mass in hypertensive patients who were never treated]. AB - A poor correlation has been found between blood pressure at rest and left ventricular mass in the course of several echocardiographic studies on hypertensive patients. The aim of this work was to determine if this finding could be the result of previous antihypertensive therapy, which had been suspended a few weeks previously in most of the studies. In addition, we tested whether blood pressure values during physical exercise correlate with the echocardiographic indices of left ventricular mass better than the values at rest. In our group of 43 patients with mild to moderate essential hypertension who had never been pharmacologically treated, the correlation between both systolic and diastolic blood pressure and left ventricular mass was poor (r = 0.41 and 0.30 respectively). This result suggests that one or more factors other than hypertension may determine the development of left ventricular hypertrophy. However, in 10 patients with left ventricular hypertrophy a more significant correlation was found between cardiac mass and diastolic pressure (r = 0.52), rather than systolic pressure (r = 0.33). This finding supports data indicating that cardiovascular risk is related more to diastolic pressure increments than to systolic pressure. As for blood pressure values during physical exercise, in our study they did not show a better predictivity of ventricular mass than the values at rest. PMID- 2978154 TI - [Neurohormonal changes which occur in heart failure and their clinical significance]. PMID- 2978155 TI - [Preparation and primary application of a new renal function imaging 99mTc-MAG3]. PMID- 2978156 TI - Comparisons between helper and suppressor T-cell induction. PMID- 2978157 TI - [Use of isotretinoin in severe juvenile acne]. PMID- 2978158 TI - [Meleda disease (mal de Meleda). Data and considerations on an indigenous caseload]. PMID- 2978159 TI - [Topical administration of hematoporphyrin derivative and red light irradiation. A therapeutic approach in psoriasis]. PMID- 2978161 TI - [Bifonazole in the treatment of dermatitis seborrheica]. PMID- 2978160 TI - [Treatment of erythrodermic psoriasis with cyclosporin]. PMID- 2978162 TI - Prevention of human helper T-cell clone activation by cyclosporin A also blocks secretion of granulocyte/macrophage colony stimulating activity. AB - Cyclosporin A (CsA) blocks stimulation and growth of alloreactive T helper cell clones (Th-TCC), even in the presence of exogenous Interleukin-2 (IL-2). To examine whether this might reflect a generalised inhibition of cytokine production by these cells, their production of granulocyte/macrophage colony stimulating factors (GM-CSF), thought not to be involved in the autocrine proliferation of the clones themselves, was investigated. Contrary to the prediction that only pathways relevant to T cell clonal expansion would be blocked by CsA, it was found that this immunosuppressive substance exerted a profound inhibitory activity on GM-CSF secretion, even in the presence of exogenous IL-2. Although a higher concentration of CsA was required to block GM CSF secretion than to block proliferation, the former was not due to toxic effects on the cells, or permanently switching off the genes for GM-CSF, since upon further cultivation of the clones without CsA, alloantigen-specific responsiveness was restored. These results therefore show that clonal human T helper cell populations are sensitive to inhibition of GM-CSF secretion by CsA. Part of the immunosuppressive mechanism of action of CsA may therefore reside in this activity, since blockade of GM-CSF secretion by T helper cells would influence functional activities of antigen presenting cells. PMID- 2978163 TI - Immunohistocytochemical localization of alpha human atrial natriuretic peptide in human atrium obtained from endomyocardial biopsy and cardiac surgery. AB - To demonstrate the localization of atrial natriuretic peptide (ANP), we were able to apply the indirect and avidin-biotin peroxidase complex (ABC) methods for immunohistocytochemical analysis on sections with routine fixation and embedding for conventional histopathological and ultrastructural examinations. Human right and/or left atrial myocardial specimens were obtained at cardiac surgery (n = 12) or by right atrial endomyocardial biopsy, employing Konno's catheter bioptome (n = 30). In both specimens, ANP-like immunoreactivity was observed mainly around the nuclei and in specific granules in atrial myocytes. These new techniques are, therefore, considered to be useful for clinical analysis employing minute human atrial specimens in various heart diseases. PMID- 2978165 TI - NSW Department of Health. Policies for developmental disability services. PMID- 2978166 TI - Back injury: prevention or cure. PMID- 2978164 TI - Increase by trifluoperazine in calcium sensitivity of myofibrils in a skinned fibre from frog skeletal muscle. AB - 1. Since it has been demonstrated that trifluoperazine (TFP) increases the affinity for Ca2+ of troponin C as well as calmodulin, the effect of TFP was examined on the Ca2+-induced tension in mechanically skinned fibres isolated from frog skeletal muscle and on Ca2+-dependent ATPase activity of myofibrils from similar frog skeletal muscle. 2. Lower concentrations of TFP increased the Ca2+ sensitivity of myofibrils without a change in the maximum tension, giving rise to a less steep tension-pCa relationship. This effect was reversible although thorough washes were necessary. The drug also enhanced myofibrillar ATPase activity, not only at low Ca2+ concentrations but also at saturating high Ca2+ concentrations. The increased affinity of troponin C for Ca2+ is difficult to accept as the sole explanation for the stimulatory effect of TFP. 3. Half of the maximum stimulating effect was obtained between 10 and 30 microM-TFP, which is similar to the reported apparent inhibition constant (Ki) for calmodulin dependent enzyme reactions. However, the stimulating effect of TFP cannot be attributed to its inhibition of calmodulin because of the finding that this effect was independent of Ca2+. Earlier published results (e.g. Klee & Vanaman, 1982) also support this conclusion. 4. Studies on myofibrillar ATPase activity suggest that the stimulating effect of TFP is not identical in its underlying action with those of caffeine and quercetin, which are also known as Ca2+ sensitizing drugs, having a similar eventual effect on tension development. 5. Higher concentrations of TFP decreased the maximum tension induced by high concentrations of Ca2+, while enhancing the tension in the presence of low concentrations of Ca2+. Analogous findings for ATPase activity were also made. TFP concentration for half the maximum depression was about 10 times higher than that for half the maximum stimulation. This suggests that different site(s) are involved in the stimulatory and inhibitory effects of TFP, although there may be some sites in common. 6. Discussion favours the stimulating effects of TFP as being caused considerably by the affected molecular interactions among myosin, actin, tropomyosin and troponin. PMID- 2978167 TI - Immunoregulation by helper T cell subsets. PMID- 2978168 TI - CR1 mediates binding of L. major metacyclic promastigotes to human macrophages. PMID- 2978169 TI - Stage-specific surface antigens during the morphogenesis of Trypanosoma cruzi: developmentally regulated expression of a glycosyl-phosphatidylinositol anchored glycoprotein of amastigotes. PMID- 2978170 TI - [Open laparoscopy]. PMID- 2978171 TI - Purification and properties of phosphofructokinase in rat submandibular gland. PMID- 2978172 TI - Effect of quinolinic acid (QUIN) on gamma-aminobutyric acid concentration in the rat brain. PMID- 2978173 TI - The ultrastructure of rat hippocampal formation in organotypic tissue culture after exposure to quinolinic acid. PMID- 2978174 TI - Molecular heterogeneity in lysosomal storage diseases. Alpha-fucosidase and N acetyl-beta-D-hexosaminidase deficiency variants. AB - The availability of specific antibodies and cDNA probes for lysosomal hydrolases has revealed unexpected heterogeneity among the human inherited lysosomal storage diseases. Using alpha-fucosidase and N-acetyl-beta-D-hexosaminidase deficiency variants as examples, it has been determined that a lysosomal hydrolase deficiency can result from DNA deletion mutations, failure to synthesize mRNA because of defective splicing, posttranslational defects in assembly, and synthesis of a precursor enzyme that is prematurely proteolytically degraded through lack of a protective protein. In some cases (fucosidosis), the different genotypes cannot be distinguished phenotypically, whereas in others (beta hexosaminidoses) the phenotypes can range from infantile neurodegeneration through juvenile motor neuron disease to adult neurodysfunction. Biochemical studies on both diseases have revealed several distinct genotypes. We show that some forms of fucosidosis result from unstable enzyme that can be stabilized by protease inhibitors, whereas partial beta-hexosaminidase deficiencies cannot be corrected by these protease inhibitors. PMID- 2978175 TI - [Bronchodilating effect of Berodual in smokers and non-smokers with chronic bronchitis]. PMID- 2978178 TI - [Blood dyscrasias]. PMID- 2978177 TI - Erythromycin and penetrating agents containing ointment in acne vulgaris therapy. AB - An ointment containing penetrating agents and erythromycin was devised. The rate of penetration of the antibiotic through the skin and the ointment efficacy in treatment of acne changes caused by Propionibacterium strains in animals were evaluated. PMID- 2978176 TI - Modulation of autologous mixed lymphocyte reaction by pharmacological agents affecting adrenergic receptor. AB - The effect of epinephrine and propranolol administration in vivo on autologous mixed lymphocyte reaction (AMLR) was investigated. AMLR inhibition by epinephrine and potentiation of AMLR after propranolol injection was found. Presumable mechanisms of these changes in the light of beta-receptor agonist and antagonist are discussed. PMID- 2978179 TI - [Bacterial endocarditis and prophylaxis in dentistry]. PMID- 2978180 TI - [Treatment of the dental patient with arterial hypertension]. PMID- 2978181 TI - [Decision making and patient management using a personal health history in a clinical dental practice]. PMID- 2978182 TI - [Polycythemia]. PMID- 2978183 TI - Alzheimer's disease and dementia. PMID- 2978184 TI - Treatment of myocardial infarction. PMID- 2978185 TI - Angioplasty in the management of coronary artery disease. PMID- 2978186 TI - [Fungal diseases of the feet in farmers]. PMID- 2978187 TI - [AIDS: the double role of the dentist]. PMID- 2978188 TI - [Prosthesis and oral lesions from metabolic diseases]. PMID- 2978189 TI - Clinical trial of ambroxol (Mucosolvan) in Sjogren's syndrome. AB - A clinical trial is described in which twelve patients with Sjogren's Syndrome (SS) were given a dose of 135 mg ambroxol (Mucosolvan) daily for eight weeks. Nine of the patients completed the trial. Three patients dropped out because they developed side effects (generalized rashes in two patients and stomatitis in one). The side effects, however, were mild and cleared off after cessation of treatment. Ambroxol improved sicca symptoms, especially ocular symptoms, in the SS patients, although lacrimal and salivary gland functions measured by Schirmer test or gum test were not changed. Furthermore, the treatment did not alter any chemical findings determined in the stimulated tear and saliva. Our results suggest that ambroxol is useful for the management of sicca symptoms in some patients with SS. PMID- 2978190 TI - [Prosthetic care of partially edentulous elderly persons]. PMID- 2978191 TI - [Perfecting metal-ceramics. Ultra light crown and bridge procedures]. PMID- 2978192 TI - [Air quality = life quality]. PMID- 2978193 TI - [Maxillo-orthopedic study model]. PMID- 2978194 TI - [The SG slide--a new solution]. PMID- 2978196 TI - Growth and precocious puberty. AB - Treatment of precocious puberty of central origin is aimed at controlling the development of sexual characteristics and improving final height. Increased growth rate is one of the major clinical symptoms, accompanied by an even more marked advance in bone age. Medroxyprogesterone acetate and cyproterone acetate have provided almost satisfactory control of pubertal characteristics, but with accompanying adrenal insufficiency. The data with regard to growth and bone maturation are contradictory. LHRH analogues have recently become available, and provide good control of gonadotrophin secretion. In a series of 21 cases (13 girls, 8 boys), a significant decrease in growth rate was achieved in both sexes with an LHRH analogue, with a significant increase in the height age/bone age ratio; control of gonadal secretions was also obtained. These results are only preliminary, but provide hope that the final height of these children will be improved. PMID- 2978195 TI - [The gingival model. An important part of dental prosthesis fabrication]. PMID- 2978197 TI - Isolation of follicular dendritic cells from human tonsils and adenoids. In vitro culture. PMID- 2978198 TI - Interactions between follicular dendritic cells and lymphoid cells. PMID- 2978199 TI - Deposits of terminal complement complex and S-protein on follicular dendritic cells in human lymphoid tissue. PMID- 2978200 TI - The leukocyte-common antigen (L-CA) family. PMID- 2978201 TI - Inhibition of splenic T lymphocytes and accessory cell interaction by anti-mac-1 (monoclonal antibody to complement receptor type 3). PMID- 2978202 TI - Suppression of immunoglobulin production by germinal centre HNK-1+ CD3+ cells. AB - Germinal centre T cells co-expressing HNK-1 antigen have little lytic activity against NK targets (K562 cells). In order to determine whether these cells regulate B cell function, they were purified from human tonsils by panning on anti-HNK-1 antibody coated Petri dishes and co-cultured with autologous and allogeneic tonsillar T and B cells in the presence of pokeweed mitogen. At the end of 7 days of culture, supernatants were assayed for immunoglobulin concentration by ELISA. A dose-dependent suppression of both IgG and IgM production was demonstrated at ratios from 1:125 to 1:16 of HNK-1+ cells to B cells, but enhancement was observed at very low ratios (less than 1:500) or ratios exceeding 1:16 in some tonsil preparations. Similar results were obtained with peripheral blood HNK-1+ cells but without enhancement in some cases at the extremes of HNK-1+ cells to B cell ratios. The suppression was not MHC restricted. These preliminary experiments indicate that germinal centre HNK-1+ cells may be intrafollicular suppressor cells. PMID- 2978203 TI - Homing receptor expression and migration of activated lymphocytes. AB - The experiments show that homing receptors are regulated in a complex fashion during initial cellular activation: Signals leading to blast formation induced either: --a decrease of homing receptor expression in the majority of blasts; - an increase of the Mel-14 expression in 20-40% of the blasts, and --a selective down-regulation in the capacity to bind to Peyer's patch HEV even under conditions, where binding to peripheral HEV is high. Submitogenic stimuli in partially activated cultures induce a rise in Mel-14 antigen expression and binding to peripheral node HEV, whereas Peyer's patch binding is unchanged or lowered. Thus, a selective and differential regulation of organ-specific homing receptors takes place under distinct activation conditions. The mucosal system related receptor is more easily down-regulated upon activation. The in vivo homing experiments indicate that mitogen activation induces one dominant migratory phenotype. Alterations in homing receptor expression seem to be associated with changes in further cellular functions leading to reduced entry into lymphatic tissues and increased localization of these cells in lung or liver. The mechanisms regulating the differential expression of organ-specific homing receptors and additional homing-relevant properties of the cells are still unknown. PMID- 2978204 TI - The migration of lymphocyte subsets from blood to lymph in the normal rat. PMID- 2978205 TI - Role of the galt contrasuppressor T cell circuit in isotype-specific immunoregulation. AB - GALT contains Tcs cells which protect IgA responses under conditions of oral tolerance. Oral immunization of erythrocyte antigen induces antigen-specific Tcs cells in Peyer's patches which predominantly support IgA responses with minor IgM and IgG responses upon adoptive transfer to tolerized mice. On the other hand, systemically-primed mice possess Tcs cells which convert oral tolerance to only IgM and IgG responses. In general, effector Tcs cells which abrogate antigen specific oral tolerance are Lyt-1+, Lyt-2-, L3T4-, I-J+ and V. villosa-adherent. Interestingly, these Tcs cells express T3 molecules which suggest the expression of a TCR which may account for their antigen specificity. PMID- 2978206 TI - Evidence for contrasuppression in patients with Crohn's disease. AB - Patients with Crohn's disease (CD) have elevated numbers of Vicia villosa agglutinin (VVA) binding cells in the peripheral blood. These cells represent a major subset of activated peripheral T cells. VVA binding T lymphocytes express either the T8 or the T4 determinant on their cell surface. In contrast in normal controls only a minor subset of peripheral T cell expresses binding sites for VVA. The majority of these cells coexpress T8. VVA binding T cells display no helper activity. Only in a subfraction of patients with CD and not in normal controls these cells mediate contrasuppressor activity for Ig and in particular for IgA. This subgroup of patients is characterized by the lack of extramucosal manifestations. It has now been shown that VVA binding T cells in their majority do not possess phenotypic features of helper inducer cells. This further supports the hypothesis of their involvement in contrasuppression. Moreover it was shown that IgA produced in the presence of VVA binding T cells is IgA1 and IgA2 (ratio 2:1) which are both modulated by VVA binding T cells. PMID- 2978207 TI - T cells and epithelial expression of HLA class II determinants in relation to putative M cells of follicle-associated epithelium in human Peyer's patches. PMID- 2978208 TI - Induction of the expression of an FcR for monomeric rat IgG2b on rat peritoneal macrophages. PMID- 2978209 TI - Human peritoneal macrophages enhance HLA-DR expression in culture and are very potent stimulators of an allogeneic mixed leucocyte reaction. PMID- 2978210 TI - Impact of IgD receptor-positive T cells on the immune response. PMID- 2978211 TI - Germinal center B cells present antigen obtained in vivo to T cells in vitro and stimulate mixed lymphocyte reactions. AB - In the present study we sought to test the ability of GC B cells to activate T cells by antigen presentation and mixed lymphocyte reactions. The first set of experiments demonstrated that GC B cells obtained three or more weeks after immunization could induce IL 2 production by T cells only by adding antigen to the cell cultures. In the next (table; see text) series, it was demonstrated that GC B cells isolated as early as 1 day and for over a week following an antigenic challenge with OVA (i.e. where they obtained antigen from the FDC-derived iccosomes in vivo), were able to stimulate high levels of IL 2 production in the absence of exogenous OVA. This response was maximal on Day 5 which corresponded precisely with the keinetics of the ultrastructural studies which document the uptake of antigen by GC B cells in vivo (see Szakal et. al, this volume). Furthermore, it was shown that the FDC-derived antigen was remarkably immunogenic when compared with exogenous antigen. The data from the MLRs demonstrated that GC B cells, which are low density cells, are potent stimulators of T cells compared to the resting, low density B cells. In addition, GC B cells even stimulated a significant syngeneic response. These results indicate that GC B cells can present antigen and interact with T cells efficiently. This is believed to be important in the germinal center reaction and in the induction and maintenance of the immune responses. PMID- 2978212 TI - Characterization of cell populations, reappearing in the mouse spleen after elimination by liposome encapsulated dichloromethylene diphosphonate: reappearance of marginal zone lymphocytes is independent of marginal zone macrophages. PMID- 2978213 TI - Epizootiological studies on porcine atrophic rhinitis. XV. Results of local immunization with a live Bordetella bronchiseptica strain in a large-scale pig herd affected by atrophic rhinitis. PMID- 2978214 TI - Ultrastructure of microgametogenesis of Eimeria stiedai in rabbits (Oryctolagus cuniculus). PMID- 2978215 TI - Studies on the virulence of pigeon paramyxovirus-1 (PMV-1). I. Changes in the virulence of pigeon PMV-1 strains isolated in Hungary upon passage in chickens, embryonated hen's eggs and pigeons. PMID- 2978216 TI - Closed abdominal surgery. PMID- 2978217 TI - Angioscopy and lasers in cardiovascular surgery: current applications and future prospects. PMID- 2978218 TI - Haemoperitoneum: an unusual complication of Fitz-Hugh Curtis syndrome. AB - A case of haemoperitoneum following minor trauma in a patient with Fitz-Hugh Curtis syndrome is presented. This is a rare complication of a condition which is common in Papua New Guinea and many other communities. Laparoscopy is the key to diagnosis of this syndrome which is often overlooked or misdiagnosed. PMID- 2978219 TI - Late rupture of knitted velour Dacron arterial prostheses. AB - Rupture of a Dacron arterial prosthesis is a most unusual complication, although it has been reported previously. Three further such cases are reported with a brief review of the relevant literature. PMID- 2978220 TI - Pathology of macrovascular disease. AB - In this chapter we have presented information on the development of large vessel damage in diabetes mellitus. A series of changes occur independent of the presence of atherosclerosis. The abnormalities include accumulation of PAS positive material, laminin, fibronectin, type IV collagen and connective tissue with lack of acid mucopolysaccharides, and deposition of calcium. It is of particular interest that accumulation of PAS-positive material and lack of acid mucopolysaccharides are recognized as the histological markers of diabetic microangiopathy. These changes are in agreement with the hypothesis of a non atherosclerotic large vessel damage, that is, diabetic macroangiopathy. From this standpoint the working hypothesis of a specific diabetic macroangiopathy should generate new ways to study the mechanism of the large vessel disease of diabetes over and above the traditional concept of classical atherosclerosis. PMID- 2978221 TI - Immunoregulatory pathways in adult responder mice. III. Establishment of a GAT specific suppressor T cell clone from GAT-tolerant responders which afferently regulates DTH responses. AB - Recent advances in the biochemical and genetic analysis of soluble immunoregulatory molecules (TsF) have been achieved via the establishment of cloned TsF-producing T cell hybridomas. However, studies on in vivo regulation of immune responses have been hampered by the lack of clonal populations of nontransformed suppressor T cells (Ts). Nonhybridoma Ts clones would allow cellular dissection of complex Ts circuits and precise analyses of Ts effector mechanisms. Our laboratory has recently demonstrated that poly(Glu60Ala30Tyr10) (GAT)-specific unresponsiveness is induced in adult responder mice tolerized via the intravenous injection of GAT-coupled syngeneic spleen cells (GAT-SP). This unresponsiveness is mediated by two antigen-specific mechanisms--nontransferable clone inhibition and induction of transferable Ts which regulate both humoral and T cell-mediated delayed-type hypersensitivity (DTH) responses. We have thus applied methodology used for the production and maintenance of antigen-specific T helper (Th) clones in an attempt to establish and characterize Ts clones mediating GAT-specific in vivo suppressive activity. Therefore, spleen cells from GAT-SP tolerant responder mice were maintained in continuous culture with soluble GAT, 10% concanavalin A-conditioned medium (IL-2), and irradiated syngeneic antigen presenting cells (APC). A stable, long-term Ts cell line (J372) was isolated by this procedure. This line and one of its clones (J372.2) suppressed the afferent (induction), but not efferent (elicitation) phase of GAT-specific DTH. In contrast, the J372.2 Ts clone had no inhibitory effect on the development of specific T cell proliferative responses. Intravenous injection of small numbers (2-5 x 10(6)) of J372.2 Ts cells resulted in significant suppression of DTH responses in GAT-primed, but not in ovalbumin- or methylated bovine serum albumin-primed recipients, demonstrating the antigen-specificity of the suppression. Intravenous injection of a GAT-specific Th clone (JTL-E1) or of a DNP-specific Th line (JTL-DNP) had no suppressive effects on GAT-specific responses suggesting that J372.2-mediated unresponsiveness is the result of active suppression, and not the result of nonspecific inhibitory effects of activated T cells. More importantly, normal GAT-specific DTH responses in recipients of the JTL-E1 Th clone (maintained in the same GAT concentration as J372.2) indicated that J372.2-mediated suppression was not due to induction of nontransferable tolerance by surface-associated GAT.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978222 TI - Cloned Lyt-1+, 2- T suppressor cells--a commentary. PMID- 2978223 TI - Adult T leukemia cells produce a lymphokine that augments interleukin 2 receptor expression. AB - Human T-cell leukemia virus (HTLV)-infected cell lines derived from adult T-cell leukemia (ATL) express constitutively the receptor for Interleukin-2 (IL-2-R) and the associated antigen (Tac antigen). In contrast, the same antigen is transiently expressed by normal T-cells only after immune stimulation. Recently, it was reported that the constitutively expressed Tac antigen on ATL cells and cell lines was not down-regulated or modulated by anti-Tac antibody. Since the antigen was modulated on normal mitogen- or alloantigen-stimulated T-cells, we postulated that the regulation of IL-2-R may be abnormal on ATL cells; the synthesis of IL-2-R is continuously stimulated in these cells. A unique HTLV/ATLV(-) cell line (YT) derived from a child with acute lymphoblastic leukemia was found to express low levels of Tac antigen that could be enhanced by various stimuli, including conditioned medium (CM) derived from normal lymphocytes, but not by lectins (PHA, Con A). Of particular interest, the exposure of YT cells to CM from ATL cell lines with helper phenotype revealed the presence of factor(s) (ATL-derived factor, ADF) that augmented the synthesis and expression of IL-2-R/Tac antigen on YT cells and promoted YT cell growth. CM from HTLV(-) leukemia cell lines lacked both IL-2-R augmenting activity and a growth promoting activity. Immunoaffinity-purified IL-2 and recombinant gamma interferon also lacked IL-2-R augmenting activity. Moreover, the physicochemical analysis with Fast protein liquid chromatography (FPLC) revealed that ADF was quite different in pI point from the IL-2-R augmenting activity in CM from normal lymphocytes. These results suggested that ADF is a unique product of HTLV(+) cells. The possible relationship between ADF production, HTLV infection, and the abnormal expression of IL-2-R is suggested, and these abnormalities may be advantageous for the leukemogenesis and abnormal growth of ATL. PMID- 2978224 TI - The in vivo behavior of T cell clones: altered migration due to loss of the lymphocyte surface homing receptor. AB - Although cloned lines of T lymphocytes have been valuable in defining the in vitro functions of well-defined cell types, they have often demonstrated relatively poor activity in vivo. One striking property of T cells clones which might affect their in vivo activity is their unusual inability to localized in lymphoid tissue as do most normal T cells. Normal lymphocyte recirculation and localization requires that lymphocytes recognize and pass through the walls of specialized high endothelial venules (HEV) as they enter into lymph nodes. We previously showed that murine T cell clones are unable to home into the peripheral lymphoid organs-lymph nodes and Peyer's patches. The inability of these cells to recognize lymph node HEV in an in vitro frozen section adherence assay suggested that the lack of lymphoid homing was due to the loss of a normal lymphocyte surface receptor for HEV. The present experiments were designed to determine: 1) the molecular mechanism responsible for the loss of normal lymphocyte migration, and 2) whether these migration and homing characteristics are irreversible features of T cell clones. Flow cytometric analysis of helper and cytolytic clones using a monoclonal antibody (MEL-14) specific for the lymphocyte homing receptor showed that they lack this surface receptor. This lack of receptor expression was confirmed by the inability to detect the antigen in detergent-solubilized extracts of surface-radiolabeled cells. Thus, the lack of homing to lymph nodes appears to be due to the loss of expression of the surface receptor which mediates the interaction between lymphocytes and HEV. When clones were rested in vitro in a nonproliferative state without stimulation by antigen or growth factors, they did not regain expression of the surface homing receptor or the ability to migrate to lymph nodes in vivo. The lack of receptor expression, therefore, is not merely associated with a rapidly proliferating state, but rather seems to be an irreversible feature of T cell clones, at least under in vitro culture conditions. T cell clones, both rested and recently restimulated, share certain features characteristic of activated T cells, as shown by recent results with MLC-stimulated T cell blasts. Both populations are large, brightly PNA-positive lymphocytes which lack expression of the MEL-14 receptor and do not home to peripheral lymphoid tissue. We propose that this PNA high, MEL-14- nonrecirculating phenotype may represent a normal phase of T cell differentiation through which many T cells pass after being activated by specific antigen.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978226 TI - The role of suppressor cells in maintaining tolerance to self molecules--a commentary. PMID- 2978225 TI - Stimulation of helper T cells and dominant suppressor T cells that recognize autologous insulin. AB - The ability to distinguish self and non-self remains a central, though incompletely understood, prescript of immunology. In the absence of disease, the organism maintains immunological unresponsiveness, or tolerance, to self proteins. Unresponsiveness also extends to certain exogenous proteins, and unresponsiveness to many of these proteins is regulated by immune response (Ir) genes encoded by the major histocompatibility complex (MHC). It has been suspected by many that there is a fundamental association between the principles that govern immunological self-tolerance and experimentally observed Ir gene control of responses to exogenous antigens. Does MHC-linked unresponsiveness to a particular antigen result from cross-reactivity between the exogenous antigen in question and self at some critical level(s)? We have approached this question through the study of in vitro antibody responses to variants of insulin. The amino acid sequence of insulin is highly conserved and murine antibody responses to heterologous insulins are controlled by H-2 linked Ir genes. We have previously demonstrated that although pork insulin fails to stimulate antibody responses in nonresponder C57BL/10 mice, it does prime helper T (Th) cells that support secondary antibody responses to pork insulin. However, dominant, pork insulin-primed suppressor T (Ts) cells normally mask this helper T cell activity. Thus, nonresponsiveness to pork insulin is controlled by highly specific suppressor T cells that prevent the expression of function, but not the priming of helper T cells. Here, we extend these studies to demonstrate that T cells from nonresponder C57BL/10 mice immunized with pork insulin have primed helper T cells and dominant suppressor T cells that cross-react with autologous insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978227 TI - Small and large B cells respond differently to T cell-derived B cell growth and differentiation factors. AB - A major problem in B cell biology is the determination of the roles played by helper T (TH) cells vs cytokines in the activation, replication, and differentiation of B lymphocytes. There is general agreement that activated B cells in cycle can replicate and terminally differentiate when provided with appropriate T cell-derived lymphokines. There is considerable controversy, however, as to whether cytokines can induce resting G0 B cells to secrete IgM. Some reports claim that TH cells are required before cytokines can act. In contrast, other reports claim that some T cell-derived supernatants (SN) have the capacity to activate resting B cells to become immunoglobulin-secreting cells. In the present study, we have examined one such SN (S26.5) as well as the EL-4 and PK 7.1 SN for their capacity to activate resting G0 B cells and a population of less dense B cells containing activated cells. These two populations, separated by Percoll density centrifugation, were characterized for size, stage in the cell cycle, and cell surface phenotype. It was shown that the most dense population contained predominantly G0 B cells, whereas the less dense population contained a subset of cells in cycle. Our studies show that neither T cell SN nor, indeed, any combination of cytokines and anti-immunoglobulins caused a major increase in the number of cells secreting IgM in the population enriched in G0 cells. In contrast, the T cell SN caused marked increases in the generation of IgM secreting cells in the population that contained a large proportion of activated cells. Limiting dilution analysis confirmed that the number of responding cells in the dense cell population was substantially lower than the number of responding cells in the less dense population. The small number of precursors in the dense B cell population may be attributed to contamination of that population with cells that have undergone activation steps in vivo. The present results, therefore, add further evidence to an existing large body of evidence that TH cells are essential for the terminal differentiation of G0 resting B cells in response to thymus-dependent antigens. PMID- 2978228 TI - Major histocompatibility complex-restricted and unrestricted activation of helper T cell lines by liposome-bound antigens. AB - Helper T lymphocytes recognize foreign antigen together with class II major histocompatibility complex (Mhc) molecules on the surface of antigen-presenting cells (APC). However, it is not known in what form soluble protein antigens are presented to T cells. The difficulty of serologically demonstrating the presence of soluble antigen on the surface of APC, the observed rapid degradation of antigen by these cells, and the finding that under special circumstances peptides of a certain protein are more antigenic that the whole molecule have led to the notion that foreign antigens must be rendered immunogenic for helper T cells by internalization, processing (probably involving enzymatic fragmentation), and redisplay on the membrane of APC in association with class II Mhc molecules. To analyze antigen recognition by helper T cells and to assess the biological significance of antigen processing, we have constructed liposomes that carry inserted class II Mhc molecules and a protein antigen coupled covalently to one of the lipid constituents of the artificial membrane. We demonstrate here that such liposomes are capable of inducing proliferative responses of long-term cultured T-cell clones, and interleukin-2 (Il-2) production by a T-cell hybridoma in an antigen-specific, Mhc-restricted fashion, in the absence of antigen presenting cells. The responses require the presence of foreign antigen and class II molecules on the same lipid vesicles. The magnitude of responses is critically dependent on the lipid composition, the density of bound antigen, and the concentration of liposomes in cell cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978230 TI - The origin of MHC-restriction of suppressor T cells--a commentary. PMID- 2978229 TI - A cloned T cell line that selectively augments antibody responses of phosphorylcholine-specific B cells bearing the T15 idiotype. AB - Helper T (Th) cells have shown to be heterogeneous both in their function and in their specificity. Several studies have demonstrated that there exist Th cells which do not appear to be specific for antigen:Ia complexes (not H-2 restricted) but interact with the target B cell via recognition of immunoglobulin idiotypic determinants. In analyzing anti-phosphorylcholine (PC) responses, such idiotype specific Th cells are characterized by 1) their ability to increase responses to phosphorylcholine by augmenting selectively the activation of T15-idiotype bearing PC-specific B cells, 2) their ability to bind specifically to T15-bearing immunoglobulin and 3) the dependence of their maturation on circulating T15 idiotype rather than Ia. Our aim is to explore further the relationship between Ia-specific and T15-specific Th cells. These studies describe the first step in comparing specificity and function of idiotype-specific Th cells to other known Th cells by the cloning of a T15-specific Lyl T cell (ThId) with properties consistent with those described for uncloned populations. The cloned ThId cells generated do not activate B cells to secrete antibody but augment anti-PC plaque forming cell (PFC) responses induced by Ia-restricted Th cells. This augmentation is seen only in the T15-bearing PFC responses even though non-T15-bearing B cells are equally accessible, and it is seen only in PC responses and not responses to TNP under identical culture conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978231 TI - Characterization of a hybridoma-derived T cell factor that promotes the production of antibodies bearing a dominant cross-reactive idiotype(s). AB - The participation of postulated subsets of T helper cells in antigen-specific antibody responses has generated both interest and controversy among immunologists. Specifically the import as well as the very existence of multiple populations of T helper cells has led to an intense search in recent years for cloned lines of such subsets that permit unambiguous classification and study. Furthermore, the means by which some of these T cells induce antibody responses may be via the elaboration of soluble factors mandating their characterization both biochemically and mechanistically. We have recently reported the existence of a T helper factor present in a 24-h Con A supernatant that specifically enhances an idiotype-bearing (Id+) response to trinitrophenol (TNP). The unique biochemical properties of this substance, namely, its capacity to bind both antigen and cross-reactive idiotype (CRI), has led to the generation of a cloned T cell hybridoma that constitutively "secretes" a factor which appears identical to the helper activity in Con A Sn. The cloned T cell hybridoma, herein designated LOP 1.4, elaborates a factor which selectively enhances the CRI+ anti TNP antibody response in vitro. The specificity of the assay employed as well as its sensitivity for detecting significant enhancement of the percent CRI+ anti TNP PFC response lent itself well as a useful vehicle for subsequent characterization of the factor. The LOP 1.4 factor, which can act at the later stages of the B cell response in a dose-dependent fashion, was characterized by affinity chromatography in order to probe the mechanism of its selective Id enhancement. The factor binds both the idiotype and the ligand for which one of the idiotype-bearing monoclonal antibodies is specific. That the factor binds idiotype and can be eluted selectively with ligand but not with noncross-reacting ligand suggests that the factor possesses separate but not independent binding sites, or alternatively, a single binding site that preferentially binds to a unique composite of antigen-idiotype. In addition, the factor bears I-J determinants, consistent with what we have previously detected on the surface of TH2-like cells. These results, collectively, suggest that the T cell hybridoma LOP 1.4 is a TH2-like cell (supporting the concept of multiple TH subsets) in light of its ability to enhance an idiotypic response to specific antigen through the production of a soluble factor that demonstrates affinity for both antigen and idiotype. In addition, like the I-J+ TH2 cell, the LOP 1.4 factor also bears I-J region determinants.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978232 TI - H-2-linked Ir gene control of VH determinant(s)-specific helper T cells. AB - The fact that helper T cells (Th) recognize antigen in the context of class II MHC antigens is well documented. T cells specific for immunoglobulin (Ig) determinants have been demonstrated as have Th cells that interact with B cells in an idiotype (Id)-restricted manner. It is still controversial whether or not such T cells recognize idiotype in an MHC-restricted fashion. In tackling this problem it is important to have a T cell population selected by the introduction of the Ig bearing the determinant(s) in question and to have both the T cell and B cell populations unbiased by prior intentional exposure to specific exogenous antigen. Thus, the likelihood of such specific antigen-induced interactions is reduced and a clearer view of the Ig-induced interaction can be obtained. With this in mind, we found that T cells from B10.D2 mice immunized with normal BALB/c serum Ig were able to stimulate the response of BALB/c B cells to sheep red blood cells (SRBC) in vitro. H-2-linked Ir gene control was revealed by the ability of these Th cells to recognize BALB/c Ig in association with H-2d (BALB/c) but not H 2b (BALB.B). Through the use of Igh congenic mice, BAB/14 and C.B20, we found the Th cells to be specific for VH (idiotypic) rather than CH (allotypic) determinants; the determinant(s) in question was apparently expressed on some BALB/c anti-SRBC antibodies since these Th cells could help anti-SRBC responses but not anti-horse or anti-burro RBC responses. This conclusion of idiotypic specificity was supported by the fact that these Th cells could be primed with either IgM or IgG from BALB/c serum, one BALB/c anti-SRBC hybridoma protein but not two others or a BALB/c IgM myeloma protein, and by the fact that absorption of the serum on SRBC prior to separation of the Ig for immunization removed the priming ability of that Ig preparation. From the use of B cell mixing experiments, it was determined that the restriction elements of H-2 complex and the appropriate Ig determinants had to be borne on the responding B cells, suggesting that direct T-B collaboration was involved in the Th cell action. Therefore, by priming with normal serum Ig we have generated Th cells which act through direct interaction with responding B cells via a VH determinant(s). In addition, unlike the findings of others using different methods of priming Id specific Th cells, these Th cells are under H-2-linked Ir gene control.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978233 TI - Varieties of idiotype-specific helper T cells--a commentary. PMID- 2978234 TI - Limiting dilution analysis of proliferating and helper T cells in the in vivo immune response to KLH: derepression of helper T cells at moderately increased frequencies. AB - While it is clear that some T cells have the capacity for almost indefinite proliferation in vitro, it is a controversial issue how much of this proliferative capacity is utilized by T cells in a response to antigen in vivo. In the framework of a strict clonal selection model the functional activities of normal and immune lymphocyte populations are essentially determined by the frequencies of antigen-specific cells which are clonally expanded after recognition of antigen. In contrast, our group has proposed a network model in which the more prominent effect of immunization is a release of antigen-specific T cells from a state of suppression (= derepression) which exists in the non immune situation and which is generated through interactions between T cells involving their antigen-specific receptors. In this model, derepression is achieved by competition of antigen with the interactions among T cells through which suppression is exerted. To test our model, we analyze in this paper how much of the immune response to keyhole limpet hemocyanin (KLH) in draining lymph nodes is accounted for by an increase in the numbers of KLH-reactive T cells or by their derepression. To this end, we immunize mice subcutaneously with KLH in CFA. For a period of 14 days after immunization draining lymph nodes are removed, and the frequencies of KLH-reactive proliferating T cells and of KLH-reactive helper T cells determined. We find that proliferating T cells increase 5 to 8 fold in frequency from day 1 to 4 after immunization (approximately 1/30,000 to approximately 1/5000) and no evidence for suppression of these T cells in the non immune situation can be obtained. In contrast, T helper cells are strongly suppressed in non-immune lymph nodes and become derepressed suddenly between days 3 and 4 following immunization. From day 0 to day 3 T helper cell frequencies are in the order of 1/14,000-1/38,000, then increase suddenly approximately 3-6 fold within 1 day from 1/16,000-1/8,000 to 1/3,000-1/5,000 with no further change until day 14. Thus, helper T cell immunity in draining lymph nodes appears to be generated by a combination of increased frequencies of specific T cells with their release from suppression. In addition, we have reasons to suspect that we overestimate the increase in T cell frequencies. We therefore think that derepression is a major factor in the response of T helper cells to antigen. PMID- 2978235 TI - Functional and biochemical evidence for the recognition of T cell receptors by monoclonal antibodies to an immunoglobulin idiotype. AB - Sharing of "idiotypes" by T and B cells with similar nominal specificities has been extensively reported in functional assays. The recent molecular characterization of T cell receptors has led to the suggestion that such idiotypic mimicries could result from "network" selection of available T cell repertoires. Alternatively, the validity of the conclusions taken from those functional assays could be questioned. We have now used an experimental system where recurrent expression of antibody idiotypes by T helper cells requires "learning" from the B cell/antibody compartment, and show here that the idiotypic determinants in question are indeed associated with T cell receptor molecules. A monoclonal antibody (F6(51)) directed to an idiotope of the TNP-binding BALB/c myeloma protein MOPC460 specifically inhibits antigen-dependent proliferation and helper activity of BALB/c anti-TNP-BALB/c helper T cells. The anti-idiotypic antibodies also induce IL-2 production by these helper cells and precipitate a surface molecule with characteristics of T cell receptor. We conclude that, in this particular system, T cell receptors and antibodies of similar nominal specificities share idiotypic determinants. PMID- 2978236 TI - Limiting dilution analysis of T cells suppressing the primary antibody response to sheep erythrocytes. AB - The immune system is often seen as an organ whose primary function is discriminating between "self" and "nonself." Theoretically, there are several possible ways it can exert such a function. Earlier, it has been discussed that clones with receptors recognizing "self-determinants" are deleted during ontogeny. However, it is now well established that the normal adult repertoire does contain T and B cells with anti-self specificity. Nevertheless, in most cases autoimmune reactions are avoided, either due to lack of stimulation or due to active control mechanisms like suppression. There are various types of suppression described in the literature, ranging from highly specific to totally nonspecific suppression. A very attractive and universal form of suppression was proposed by Jerne in his network hypothesis: in the nonimmune state, cells of the immune system communicate with each other via interactions of their specific receptors and thus form a self-suppressive network. This paper describes the attempt to estimate frequencies of suppressor T (Ts) cells existing in the normal nonimmunized mouse. Ts cells are defined functionally in a suppressor assay, i.e., by suppression of the in vitro primary immune response of spleen cells to sheep erythrocytes. The experimental procedure involves limiting dilution of T cells into the suppressor assay followed by a quantitative analysis of the antibody responses (PFC assay or ELISA) and Poisson statistics. Several separate "peaks" of suppression are observed, depending on the number of T cells in the assay. Varying from experiment to experiment, these peaks reach maxima of suppression ranging from 20 to 80%. Low numbers of T cells are especially efficient in suppression, being themselves counterregulated at higher cell numbers. With increasing T cell numbers, suppression will appear and disappear again several times--a phenomenon already described by us for other functional T cell populations [reviewed in Eichmann et al. (1983): Springer's Sem. Immunopathol. 6:7].(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978237 TI - Sporadic idiotypic cross-reactivities between antibodies and T helper cells: one example of aleatory expression of T cell idiotypes. AB - Idiotypic cross-reactivities between T and B cell receptors have been taken in the past to suggest VH-gene expression by both types of lymphocytes. More recently, after the molecular characterization of TcR, those observations were reinterpreted to indicate idiotypic network regulation, operating to select cross reactive idiotypes in both B and T cell compartments. In support of these views, it has been shown that T cell expression of idiotypes is controlled by IgH-linked genes and markedly altered in B cell-deprived mice, and that T cells "learn" idiotype expression from the B cell compartment in the first few weeks of life. In most of these studies, aleatory idiotype cross-reactivities have not been sufficiently considered. Given the large diversity of antibodies and TcR, and the degeneracy of antibody-ligand interactions, it could be expected that a (monoclonal) anti-idiotypic reagent prepared against an antibody idiotope will always "cross-react" with some TcR variable regions. If these will be "major" T cell clonotypes, situations of idiotype sharing by B and T cells can be found which indicate neither VH-gene expression by T cells nor idiotypic network regulation. We report here one example of such aleatory expression of antibody idiotypes by T lymphocytes. A monoclonal anti-idiotypic antibody directed to anti TNP antibodies of BALB/c mice specifically inhibits the growth and effector activity of C57BL/6 anti-NP-self helper T cells, while failing to interact with either BALB/c anti-NP-self or C57BL/6 anti-TNP-self helper cells. The clonotypic nature of these interactions could also be demonstrated by the specific induction of IL-2 production in the appropriate helper T cells by the anti-idiotypic antibody.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978238 TI - The role of the murine L3T4 molecule in T cell activation: differential effects of anti-L3T4 on activation by monoclonal anti-receptor antibodies. AB - MHC restricted T cells can be divided into two subsets based on the mutually exclusive expression of the cell surface differentiation antigens L3T4 and Lyt-2 in the mouse. Expression of the L3T4 marker is correlated most strictly with recognition of foreign antigen in association with self class II MHC molecules, or Ia molecules. Less stringently correlated with L3T4 expression is the recognition of unmodified self or non-self Ia molecules. Finally, expression of L3T4 is also correlated with certain functional properties, although this correlation is even less stringent. The major correlation for function is between L3T4 and the ability to activate B cells. These correlations have led to the hypothesis that L3T4 recognizes Ia molecules, and plays a role in increasing the affinity of T cell:Ia bearing cell interactions. This hypothesis is bolstered by the finding that anti-L3T4 antibody blocks such interactions. Recently, we and others proposed a second effect of cross-linking L3T4 molecules, namely negative signalling. We further proposed that the natural ligand for L3T4 is Ia molecules, and that Ia-driven cross-linking of L3T4 molecules on the T cell in the absence of receptor aggregation would lead to off signalling to the T cell and separation of cell conjugates. To better understand the role of the L3T4 molecule in T cell activation, we have examined the effect of several anti-L3T4 antibodies on stimulation of a cloned line of helper T cells by a panel of monoclonal antibodies directed at what appear to be different epitopes on the T cell receptor. Unlike previous analyses of stimulation of helper T cells with anti-T cell receptor antibodies, we observe differential effects of anti-L3T4 on T cell activation by anti-receptor antibodies, the effect of anti-L3T4 depending on the characteristics of the anti-receptor antibody. This result suggests that L3T4 is intimately associated with the T cell receptor, and may thus play a critical role in T cell specificity as part of the antigen:Ia recognition complex. This proposed role is in keeping with the very strong correlation between L3T4 expression and recognition of self class II MHC molecules. While these studies do not provide definitive evidence for a physical association between L3T4 and the T cell receptor, they do place certain constraints on current models and suggest new possibilities for understanding T cell recognition and development.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978239 TI - Direct receptor:receptor interactions between T and B lymphocytes: idiotypic restriction in the antibody response to a cloned helper T cell receptor. AB - The concept of an immunological network includes the possibility of interactions between receptors on T and B lymphocytes, and such interactions, should they occur, might be expected to influence the repertoire of receptors in each set of cells. Indeed, B cell idiotype specific helper T cells, both MHC-restricted and MHC-unrestricted, have been reported and have been shown to influence the expression of the B cell repertoire. Likewise, it has been reported that B cells may influence the specificity of both regulatory and MHC-restricted T cells. However, interactions between receptors on cloned, MHC-restricted helper T cells and B cells have been difficult to document. Recently, we have taken advantage of an unusual cloned helper T cell line to demonstrate that anti-T cell receptor antibody is produced by direct receptor:receptor interactions between T and B lymphocytes, and that these interactions are not MHC restricted. However, these earlier studies did not address the question of whether such interactions led to activation of B cells expressing multiple distinct antibodies, or whether direct T cell receptor:B cell receptor interactions would lead to an idiotypically restricted B cell response. To address this question, we have now examined both monoclonal and polyclonal responses to the receptor of a conventional, MHC restricted cloned T cell line, and have shown that these responses are of limited idiotype heterogeneity. Indeed, about 60% of antibodies produced to the receptor of this cloned line share idiotypic determinants, and appear to recognize a single epitope on the receptor. Idiotypically unrelated anti-receptor antibodies, although still specific for the cloned line, recognize what appears to be a distinct epitope on the receptor. These data suggest several conclusions. First, they demonstrate further that direct receptor:receptor interactions between helper T cells and B cells can occur, and can be mutually stimulatory for the two cell types. Second, as shown previously, such interactions are not MHC restricted. Third, such interactions can lead to an idiotypically restricted B cell response. Finally, it is interesting to compared these results with those of other investigators studying idiotype-specific helper T cells. As the cloned line used in this study is a conventional, MHC-restricted, antigen specific helper T cell bearing an alpha:beta heterodimeric receptor complex, and as its interaction with B cells is MHC unrestricted and leads to idiotypically restricted antibody responses, one might propose that such cells are candidates for a clone of an idiotype-specific helper.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978240 TI - A helper T cell clone produces an antigen-specific molecule (T-ABM) which functions in the induction of suppression. AB - Among Ly-1+,2-T cells there appears to be two independent modes of antigen recognition. Helper and cytotoxic Ly-1 T cells recognize antigen only in the context of I region products whereas regulatory T cells, such as T suppressor inducer cells, produce antigen-specific, antigen-binding molecules (T-ABM). These T-ABM often have been found to form a part of biologically active, antigen specific regulatory factors. A number of environmental conditions effect whether a foreign antigen will produce a positive response leading to immunity or a negative one leading to tolerance. Many of the conditions which favor the induction of suppressor T cells simultaneously preclude the proper interaction of antigen presenting cells with helper T cells. This parallel led us to ask whether helper T cells perform at least two, apparently opposite functions: a) under conditions favoring immunity helper T cells produce lymphokines to activate immune effector cells, and b) under conditions favoring suppression they produce molecules which function in suppressor cell induction. Therefore, this question relates to the mechanisms by which an immune response is switched into either a positive (help) or negative (suppressive) track. In addition, it begins to address the relationship between the different modes of antigen recognition exhibited by helper T cells vs. T suppressor inducer cells (see above). To explore this problem we employed an antigen-specific, I-Ak restricted helper T cell clone as the purest available source of helper T cells. We presented antigen to the cloned T cells under conditions which favor suppression rather than help (for example, by ultraviolet irradiation of the antigen-presenting cells) and collected supernatants 48 hrs later. The supernatants were then examined for activity in a functional assay for antigen-specific suppressor factors. Our results indicate that under conditions favoring suppression, a T-ABM was produced which functioned in the antigen-specific induction of suppression in vitro. The T ABM had the same antigen specificity as that exhibited by the helper T cell and was therefore probably derived from the clone. This observation introduces the possibility that the interaction between antigen-presenting cells and helper T cells is a crucial decision point in the immune response which can lead to either immunity or suppression. The latter would be achieved through the production, by helper T cells, of an antigen-specific component of T suppressor inducer factor (i.e., the T-ABM). The possible relationship between T-ABMs and the T cell receptor is discussed. PMID- 2978241 TI - Induction of suppression by a murine nonspecific suppressor-inducer cell line (M1 A5). III. Partial purification of the suppressor cell-inducing factors. AB - The cause of the state of anergy which often accompanies advanced tumor growth has been attributed to several factors, including nonspecific suppressor cells capable of suppressing humoral and cell-mediated responses. Induction of suppressor cells in tumor-bearing hosts has been attributed to the release of immunoregulatory factors by the tumor, as well as by the host's own lymphoid cells. Several such "suppressor cell-inducing factors" have been described. However, in only a few cases has the inducing material been characterized. We have recently reported the existence of a suppressor cell inducing factor (SIF) in the 18 hr culture supernatant of spleen cells from mice bearing an advanced M 1 fibrosarcoma. Our aim was to characterize SIF, both biochemically and mechanistically. In order to purify the inducing factor, as well as to dissect the cellular and molecular events that occur as a result of suppressor cell activation, we isolated the M1-A5 cell line from the spleen cells of a mouse bearing an advanced M-1 fibrosarcoma. Induction of suppressor cells by supernatants from M1-A5 cells closely resembled the situation seen with the whole spleen cells. In both cases: 1) an inducing factor with an estimated Mr less than 12 kDa was found, and 2) the release, but not the effector function, of the inducing factor was prostaglandin-dependent. The 18 hr culture supernatant of M1 A5 cells was used as the source of the inducing factor and suppressor cells were activated by exposure of normal spleen cells to the inducing factor for 4 hr. The nature of the inducing factor was investigated by subjecting the M1-A5 culture supernatant to molecular weight sieving on Sephadex G-100 medium. The results showed that M1-A5 supernatants contained a high- (Mr 70 kDa) and a low- (Mr 5.5 kDa) molecular weight factor, which were termed SIF alpha and SIF beta, respectively. SIF alpha and SIF beta were further purified by ion exchange chromatography. SIF beta bound to QAE-Sephadex A-25 and was eluted in a single peak. However, SIF alpha displayed heterogeneity with respect to binding to DEAE Sephacel, as SIF activity was seen in the void volume as well as in fractions eluted from the anionic exchanger. Mechanistically, we demonstrate that the induction of suppressor cells by SIF alpha and SIF beta is genetically non restricted. In addition, we show that SIF alpha and SIF beta suppress the in vivo antibody response to sheep red blood cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978242 TI - Possible involvement of Ly-1 B cells in the effector phase of IgG suppression mediated by suppressor T cell factor. AB - The effector phase of IgG suppression mediated by an antigen-specific suppressor T cell factor (TsF) was studied. The monoclonal TsF of an inducer type derived from a KLH-specific suppressor T cell hybridoma (34S-704) suppressed IgG response mounted by DNP-primed B cells (anti-Thy-1 treated spleen cells) and KLH-specific cloned helper T cells only in the presence of unprimed Thy-1-, Ly-1+, I-J+, Ig+ cells. The addition of naive Ly-1+ cells to the culture of DNP-primed Ly-1/Thy-1 depleted B cells and KLH-Th clones augments anti-DNP IgG responses, and KLH-TsF suppressed the enhanced parts of IgG responses. The Ly-1+ cell seems to be a target of TsF, because naive spleen cells treated either with anti-Ly-1 or with anti-MIg failed to absorb TsF, whereas TsF activity was absorbed with whole spleen cells and anti-Thy-1 treated spleen cells. The functional role of Ly-1 B cells and possible mechanisms in the effector phase of TsF-mediated IgG suppression will be discussed. PMID- 2978243 TI - Activation of help and contrasuppression as essential prerequisites for immune response. AB - The network theory proposes the immune system as a self-centered defense mechanism, which continuously maintains a steady state of activity via idiotypic anti-idiotypic interactions. In line with this hypothesis, the steady state of the immune system has been described to represent a status of suppressed activity, response being manifested by release from suppression. There is evidence that release from suppression is initiated by activation of contrasuppressor cells (TCS), which either transfer a state of resistance towards suppression on helper T-cells (TH) or interact directly with suppressor T-cells (TS). In the latter case, it was postulated that the nominal antigen of contrasuppressor T-cells are antibodies and that TS and TCS interact one with the other via idiotypic-anti-idiotypic structures. The present report examined the role of TCS in initiation of response and proved that the responding state requires activation of TH as well as TCS. While antigenic stimulation (TNP) resulted in concomitant activation of TH and TCS, it was possible to dissect these two prerequisites for response by application of a monoclonal anti-TNP antibody (AB) carrying a recurrent idiotype (Sp6) followed by application of a subimmunogenic dose of the nominal antigen. Clonal analysis of regulatory cells via limiting dilution (LD) cultures revealed that a subimmunogenic dose of antigen led to activation of help but failed to activate TCS. On the other hand application of AB, which did not initiate response, resulted in activation of TCS, but not in activation of TH, i.e., these were not released from suppression. However, consecutive activation of TCS via AB and of help via a subimmunogenic dose of antigen moved the immune system into the responding state. Activation of TCS via AB was found to be independent from the mode of application, i.e., free AB, AB coupled to syngeneic cells, or AB coupled to the nominal antigen, although most straightforward results were obtained with free AB, since syngeneic cells exerted an additional suppressive effect, and AB-antigen (TNP) conjugates activated TNP-specific TH as well as TCS. Furthermore, AB-induced activation of TCS was independent of the recipient's age. Yet, the effect was most pronounced when AB were applied neonatally, i.e., after a hyporesponsive stage, animals were hyperreactive towards the nominal antigen, and this hyperreactivity was initiated by expansion of TCS. This extends the notion of high idiotypic connectivity in the neonatal period to the level of regulatory T-cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978244 TI - The role of suppressor T cells in the expression of immune response gene function. AB - Mechanisms underlying major histocompatibility complex (MHC)-linked immune response (Ir) gene regulation of immune responses have been the subject of considerable interest and debate in recent years. Two general mechanisms have been proposed to account for antigen-specific, Ir gene-mediated unresponsiveness. In one, defective antigen presentation resulted from the failure of processed nominal antigen and Ia antigen to associate on the antigen presenting cell membrane in a manner sufficient for helper T cell (Th cell) activation. By contrast, it has been proposed that selected Th cell clones were deleted from the repertoire during ontogeny or otherwise rendered unresponsive to the antigen-Ia complex, i.e., functionally deleted. Either of these mechanisms would account for the deficient activation of antigen-specific, Th cells observed in genetic low or nonresponder mice. In addition, the failure of mice to respond to certain antigens under Ir gene control has been attributed to the activation of specific suppressor T (Ts) cells. The latter mechanism might be considered a corollary or subset of the clonal deletion model. However, an important distinction exists. In the case of active, Ts cell-mediated Ir gene regulation, genetic low responder animals should retain the capacity for antigen-induced activation of Th cells, or Th cell activity should be demonstrable in these mice. In this communication, experiments are described which are designed to evaluate the possibility that active Ts cell-mediated regulatory mechanisms were of general importance in mediating Ir gene-related unresponsiveness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978245 TI - Ir gene regulation: past failures to present cogent mechanisms and to delete diverting oversimplifications--a commentary. PMID- 2978246 TI - A comparison of salbutamol and ipratropium in chronic bronchitis and chronic airflow limitation. PMID- 2978247 TI - [Genetic monitoring of inbred rats by electrophoresis technique]. PMID- 2978248 TI - [Diagnostic value of ratio of serum hCG level to CSF in the brain metastasis of choriocarcinoma]. PMID- 2978249 TI - [Expression of intermediate filaments in cultured cells]. PMID- 2978250 TI - [Experimental study of atherosclerosis. I. Anti-atherosclerotic actions of Chinese herb effective component 8501]. PMID- 2978251 TI - [A formula method for measuring the oscillatory potentials of electroretinogram]. PMID- 2978252 TI - [Studies on anatomy, roentgenology and mechanics of calcar femorale and their clinical significance]. PMID- 2978253 TI - [Preliminary observation on plasma level of 5-HT, TXB2 and 6-keto-PGF1 alpha in TEPH]. PMID- 2978254 TI - [Cyclosporin A inhibits arachidonic acid metabolism in cultured cells]. PMID- 2978255 TI - [Generation and identify of a panel of monoclonal antibodies against carcinoma of the lung]. PMID- 2978256 TI - [Long-term follow-up on change of antithyroid antibody in chronic lymphocytic thyroiditis]. PMID- 2978257 TI - [Treatment of rheumatoid arthritis with tripterygium wilfordii hook. I. Effect on secretion of total IgM and IgM-RF by peripheral blood mononuclear cells (PBMC)]. PMID- 2978258 TI - [Establishment of cell-sorting technique with flowcytometer]. PMID- 2978259 TI - [Study on short-term clinical trial of R-77-3 [3-(4-cyclo-pentyl-l-piperazinyl) imino methyl rifamycin SV] by mouse foot-pad technique]. PMID- 2978260 TI - [Ultrastructural studies on differentiation of human leukemic cell induced by retinoic acid and harringtonine]. PMID- 2978261 TI - [Primary investigation of drug-induced lupus]. PMID- 2978262 TI - Prolactin levels and pituitary enlargement in hormone-treated male-to-female transsexuals. AB - PRL levels were evaluated during long-term treatment with cyproterone acetate 100 mg and ethinyloestradiol 100 micrograms/day orally or depot-oestrogens in 214 male-to-female transsexuals. PRL levels increased above normal in all subjects (normal less than 300 mU/l). In 46 (21.4%) subjects PRL levels rose to greater than 1000 mU/l. The incidence of PRL levels greater than 1000 mU/l was 3.7-7.2% per treatment year. Grossly elevated PRL levels were associated with high doses of oestrogens (P less than 0.05) and advanced age at the start of treatment (P less than 0.05). In 23 subjects PRL levels greater than 1000 mU/l decreased by more than 50% spontaneously (n = 5) or after dose reduction (n = 18). In five of the subgroup of 15 subjects with persistent PRL levels greater than 1000 mU/l enlargement of the pituitary gland was shown by CT-scanning. These data suggest that the lowest possible oestrogen dose and lifelong follow-up of hormone-treated male-to-female transsexuals is essential. PMID- 2978263 TI - The reversal of the metastatic phenotype by gene transfer. AB - When studying the function of MHC-restricted immune responses in controlling metastatic growth we discovered that highly metastatic clones of mouse tumours express the H-2D but lack expression of the H-2K gene of the MHC system. The de novo expression of the H-2K antigen, after H-2K gene transfection, resulted in the reversal of a metastatic to a non-metastatic phenotype. This reversal was causally related to the acquisition of H-2K-restricted immunogenic properties. Immunization with H-2K-transfected cells, after surgical removal of the local tumour, abolished or significantly reduced the growth of metastases. We subsequently observed that H-2K expression is correlated with expression of the c fos oncogenes. Transfection of H-2K-negative cells with v-fos or c-fos genes resulted in the expression of H-2K. Our studies suggest that one of the main functions of the c-fos proto-oncogenes is control of the expression of the MHC genes. Searching for additional molecular properties which characterize the metastatic phenotype, we observed that the metastatic clones of each of our lung metastasizing tumours expresses an fms-related oncogene. This was correlated with a membrane-bound tyrosine kinase, which has the properties of growth factor receptors. We examine the possibility that our fms-like gene codes for this protein kinase, which represents a receptor for a local growth factor that controls metastatic growth in the lung. PMID- 2978264 TI - [Incidence of onychomycoses in ungual pathology]. PMID- 2978265 TI - [Nevus sebaceous]. PMID- 2978266 TI - [Topical antiandrogen therapy in seborrheic blepharitis]. PMID- 2978267 TI - [Facts about AIDS for the dental professional]. PMID- 2978268 TI - [The effect of topical stimulation on periodontal structure in Streptozotocin induced diabetic rats. 1. The experiment in conventional rats]. PMID- 2978269 TI - [The effect of topical stimulation on periodontal structure in Streptozotocin induced diabetic rats. 2. The experiment in germfree rats]. PMID- 2978270 TI - Generation of self-macrophage-toxic non-T cells in the MHC-homozygous F1 spleen cells co-cultured with parental cells: possible involvements of host cells in impaired immunity in GVH disease. AB - Simplified-in vitro system was developed to examine the contribution of host's cells in graft-versus-host (GVH)-disease-associated immunodeficiencies. In analogy with major histocompatibility complex (MHC)-matched GVH-reaction, (BALB/c x DBA/2)F1 (H-2d) hybrid spleen cells were co-cultured with irradiated BALB/c (H 2d) spleen cells, so that cellular activities to be generated are ascribable to F1 cells. In vitro development of anti-allo-specific cytotoxic T cells of the F1 origin was dramatically suppressed by coexistence of the irradiated parental cells and by the addition of F1 cells precultured once with the parental cells, suggesting the generation of suppressor cells in the F1 (host) cells activated by the parental cells. Thus generated suppressor cells are Thy.1-, weakly or nonadherent and radiosensitive. Interestingly, in the same reactions there also developed Thy.1- cytotoxic cells for autologous macrophage targets. An involvement in immunodeficiencies in GVH disease of the host-derived cytotoxic and/or immunosuppressive, non-T cells was discussed. PMID- 2978271 TI - Pulmonary involvement in schistosomiasis mansoni. AB - The post-treatment pulmonary alterations were evaluated in patients (Study 1) and in mice (Study 2) infected with Schistosoma mansoni. Study 1: the patients were examined pre and post-treatment (with ora oxamniquine) and the following exams were performed: sputum for eosinophils and chest x-ray. Study 2: four groups of mice (total = 64) were studied; Group I (infected and treated with oxamniquine); II (infected and not treated); III (not infected and treated) and IV (not infected and not treated). All were x-rayed to check for pulmonary abnormalities pre and post-treatment and lung specimens were studied by optical microscopy and immunofluorescence. We have found abnormalities in the parameters checked in both studies and the results suggest an immunological reaction, probably due to deposition of immune complexes in the lungs, with subsequent activation of the complement system. The experimental study showed that the alterations are not dependent of the presence of eggs and/or worms of S. mansoni in the lungs, thus corroborating the hypothesis of deposition of circulating material. PMID- 2978272 TI - [Reactions of tonically active motor neurons to the stimulation of different afferents]. PMID- 2978273 TI - Renal disease: a dental perspective. PMID- 2978274 TI - [Program of vaccination against hepatitis B in Latium]. PMID- 2978276 TI - [Acquired immunodeficiency syndrome (AIDS): review]. PMID- 2978275 TI - [Effect of the method of inhalation of Berodual on bronchodilation in patients over 60 years of age with diffuse bronchial obstruction]. PMID- 2978277 TI - Clinical and histologic observations of glass ionomer-silver cermet restorations in six human primary molars. PMID- 2978278 TI - Cell membrane Ca2+/Mg2+ ATPase. PMID- 2978279 TI - [AIDS: an opinion survey of dentists in the Republic of South Africa]. PMID- 2978280 TI - Dehydroepiandrosterone sulfate (DHEA-S) and DHEA-S-like compounds in fibrocystic disease of the breast. AB - We assayed Type 1 (high K+) and Type 2 (high Na+) human breast cyst fluids for DHEA-S. When an antibody specific for the 3-sulfoconjugate end of DHEA-S was used, Type 1 cyst fluids (n = 18) showed a content of 114 +/- 68 micrograms/mL (mean +/- sigma) and Type 2 cyst fluids (n = 14) of 35 +/- 17 micrograms/mL (P less than 0.01). Using an antibody specific for the D-ring, the results were 151 +/- 91 micrograms/mL and 51 +/- 32 micrograms/mL, respectively (P less than 0.01). The apparent concentrations of DHEA-S were statistically different, even though both assays gave equal results in serum from normal adults. The presence of other compounds in individual cyst fluid samples was examined by extraction and chromatography. DHEA-S immunoreactivity was found in both early and late eluting fractions in Type 1 cyst fluids and in late eluting fractions from Type 2 cyst fluids. Only the late eluting fraction from Type 2 fluids had approximately equal immunoreactivity with both antibodies. In addition to authentic DHEA-S, breast cyst fluids contain other materials that react with DHEA-S antibodies. Radioimmunoassays for DHEA-S in cyst fluid must be specifically validated because of the presence of these compounds. PMID- 2978281 TI - A new reversed phase, paired ion thin-layer chromatographic method for steroid sulfate separations. AB - The sulfoconjugated steroids estrone sulfate (ES) and dehydroepiandrosterone sulfate (DS) were separated in the reversed phase mode on polyamide-coated TLC plates. Baseline resolution was obtained between tritiated ES and DS standards when run with a mobile phase of 20% acetonitrile in 5mM aqueous triethylamine, triethanolamine, tris-hydroxymethylaminomethane, tributylamine or ammonia. ES and DS showed no mobility in the absence of an ion-pair reagent. The radioactive peaks were detected and integrated non-destructively by scanning. Quantitation was confirmed by elution of cut-out peak areas and liquid scintillation counting. Similar results were obtained with washed ethanol extracts of serum labeled with tritiated ES and DS. The extracts were defatted on the plate with hexane: ethyl acetate (1:1) prior to the reversed phase development. PMID- 2978282 TI - Perinatal management of the fetus with an abdominal wall defect. AB - The antenatal diagnosis of abdominal wall defects has allowed improved perinatal management. For fetuses with associated anomalies, the options of elective termination or minimal intervention can be offered. Our ability to predict the extent of bowel damage in gastroschisis based on the ultrasound findings enables us to offer early delivery to those fetuses who are at high risk. The data are not clear at the present time whether cesarean section offers any advantage. These fetuses should, however, be delivered at a center which is capable of providing high level medical and surgical care to these potentially ill infants. Initial resuscitation of these neonates requires early insertion of an intravenous line and a nasogastric tube, the administration of antibiotics, sterile coverage of the eviscerated bowel, and careful attention to temperature instability. Neonates with gastroschisis should be operated on as soon as they are stable, whereas infants with omphalocele can be investigated for associated anomalies prior to surgery. Primary fascial closure is performed whenever possible. Where this is not possible, a staged repair using a silastic chimney achieves closure within 3-6 days. Skin coverage alone or nonoperative management is reserved for the few cases with giant omphalocele, associated anomalies, or poor operative risk. Decisions about primary versus delayed closure, while usually dictated by clinical judgement, can be aided by indirect measurement of intraabdominal pressure. Postoperative ventilation, and consideration of long term nutritional needs, are also important parts of the perioperative management. PMID- 2978284 TI - [The role of IL-2 and Th in pathogenesis of non-Hodgkin's lymphoma (NHL)]. PMID- 2978283 TI - [Effects of phenobarbital and phenytoin on the growth morphology and some ATPase activities of rat brain astrocytes derived from normal newborn cerebrum and C6 glioma cells in culture]. PMID- 2978285 TI - [Study of pancreatic islet's reconstruction and beta cells regeneration of experimental diabetes]. PMID- 2978286 TI - Contrasting plasma atrial natriuretic factor concentrations during comparable natriuresis with infusions of atrial natriuretic factor and saline in normal man. AB - 1. To explore the role of atrial natriuretic factor (ANF) in the response to an acute saline load, we compared plasma hormone and urinary electrolyte interrelationships during administration of ANF and saline. Plasma concentrations of ANF, renin, angiotensin II and aldosterone, together with urine volume and electrolytes, were measured during infusions of placebo, ANF [alpha-human ANF (99 126)] and 0.9% (w/v) NaCl solution in normal subjects under standardized conditions of diet and posture. 2. Saline loading and ANF infusions initially induced similar natriuresis and suppression of renin-angiotensin-aldosterone system activity in association with markedly disparate values of plasma ANF. Plasma ANF levels rose to two- and eight-fold placebo values with saline and ANF, respectively (P less than 0.001). Conversely, in the period after infusion plasma ANF values were similar while natriuresis differed significantly. Peak natriuresis lagged behind peak plasma ANF values with both stimuli. 3. ANF, but not saline, enhanced urinary excretion of calcium and magnesium. Saline, but not ANF, caused increased kaliuresis. 4. The data suggest that ANF makes only a minor contribution to natriuresis induced by saline challenge, although full confirmation of this point requires quantification of end-organ responses to endogenous ANF in the face of changing arterial pressure and circulating volume. PMID- 2978287 TI - Atrial natriuretic factor increases after a protein meal in man. AB - 1. The renal function changes induced by dietary protein are thought to result from the activity of hormonal factors that remain as yet undefined. Since a meat meal and high dose atrial natriuretic factor (ANF) infusions have similar effects on glomerular filtration rate, natriuresis and kaliuresis, we decided to investigate the possibility that a protein meal could stimulate ANF activity. 2. We studied 10 normal volunteers who had a fixed protein and sodium intake for 7 days before the experiments. The subjects received a meat meal (1-1.5 g of protein/kg) and, on a separate occasion, a carbohydrate meal that had a similar caloric, sodium and potassium content. Diuresis was stimulated with water ingestion, and urine collections were obtained before the meals (baseline) and after the meals for a period of 3 h. Blood samples were obtained 30 min and 5 min before the meals and every hour for 3 h in the period after the meal. 3. The protein meal, but not the carbohydrate meal, was associated with parallel increments in plasma immunoreactive ANF (i-ANF), natriuresis, kaliuresis and glomerular filtration rate (estimated from creatinine clearances) which reached peak values 2-3 h after the meal. The mean increment of plasma i-ANF after the protein meal represented a twofold increase over baseline levels. 4. We conclude that ANF may participate in the physiological response to an oral protein load. PMID- 2978288 TI - raf family serine/threonine protein kinases in mitogen signal transduction. PMID- 2978289 TI - Topical corticosteroids for skin disorders in infants and children. AB - Topical corticosteroids are an essential part of the treatment regimen of children with atopic eczema and a number of other inflammatory dermatoses, such as infantile seborrhoeic eczema, pompholyx and contact dermatitis. A special form of treatment, the 'wet wrap technique, has been found to be effective in treating children with acute erythrodermic eczema in an inpatient setting. Therapy usually lasts for 3 to 5 days and is useful as crisis intervention therapy. On the other hand, in some conditions, for example pityriasis alba, perioral eczema or juvenile plantar dermatosis, therapy with these agents is generally not required or may not be useful. It is important to remember that care must be taken when a topical corticosteroid is prescribed in children, since this age group is particularly at risk of developing side effects. Thus, the patient should be monitored closely, and potent agents should be avoided. PMID- 2978290 TI - [Necrotizing fasciitis of the abdominal wall]. PMID- 2978291 TI - Effect of gossypol acetic acid on spermatozoal ATPases activity and fertility in chicken. PMID- 2978292 TI - Role of adrenal in maintenance of spermatogenesis in rats. PMID- 2978293 TI - Comparative double-tracer whole-body autoradiography: uptake of 11C-, 18F- and 3H labeled compounds in rat tumors. AB - The uptake of various labeled compounds by tumors was studied by double-tracer whole-body autoradiography (DTWBA) in rats. Each animal carried two types of tumors: mammary carcinomas and the Walker 256 carcinosarcomas. The markers used were [18F]- and [3H]fluorodeoxyglucose (glucose utilization), [3H]thymidine (cell proliferation), [11C]methionine (amino acid metabolism) and [11C]- and [3H]toremifene (estrogen-receptor-avid agents). In each experiment, the distribution of a substance labeled with short-lived radionuclide (11C or 18F) was compared with that of another substance labeled with a long-lived nuclide (3H). Quantification was done by combining computerized image analysis of the autoradiograms with liquid scintillation counting of punched tissue pieces obtained from the cryosections. The relationships between the uptakes of the various radiopharmaceuticals were recorded in tumors and normal tissues. The dynamics of [18F]fluorodeoxyglucose and [11C]methionine were determined in tumors and some selected tissues by positron emission tomography (PET). The uptake rate between fluorodeoxyglucose and thymidine in the mammary tumor was five times higher than the ratio in the Walker tumor. The corresponding figure for FDG/methionine was four times. Thymidine, compared with methionine, was twice as efficient. Thus, the mammary tumors were best imaged with FDG or thymidine. The non-steroid antiestrogen toremifene was taken up in very low amounts by these tumors. By DTWBA, experimental tumors may serve as their own control. PMID- 2978295 TI - Evaluation and management of the dental patient with renal failure, renal transplant or on renal dialysis. PMID- 2978294 TI - Muscle-spindle distribution in relation to the fibre-type composition of masseter in mammals. AB - The various parts of the masseter muscle complex (pars superficialis, pars profunda, zygomaticomandibularis, maxillomandibularis) in the rat, guinea-pig, rabbit, cat and macaque monkey were examined to discover whether they showed any relationship between the distribution of muscle spindles and extrafusal fibre types. Intrafusal (spindle) and extrafusal fibre types in masseter were compared with those in limb muscles and were identified by a combination of standard histochemical methods and indirect immunoperoxidase staining with antibodies specific for the various isoforms of myosin characteristic of fibre types in mammalian muscle. In general, the fibre-type properties of intrafusal fibres in masseter resembled those in limb muscle spindles, but the extrafusal fibre-type composition was unlike that in most limb muscles. In the rat masseter, most of the spindles were clustered together in a few very restricted areas. Extensive fusion of the external capsules of adjacent spindles, resulting in the formation of giant spindles, was seen in the cat and monkey masseter; this was sometimes accompanied by the enclosure of extrafusal fibres within the fused spindles. Common to all species, but strongest of all in the rat, was a close association between the distributions of muscle spindles and extrafusal Type I (slow twitch) fibres within the masseter complex. Muscle spindles and Type I fibres were either absent or rarest in the superficial part of masseter, but were most common in the deep layer (pars profunda) or zygomaticomandibularis. The functional significance of these observations is discussed. PMID- 2978296 TI - Beta-subunit of ATP-synthase: a useful marker for studying the phylogenetic relationship of eubacteria. AB - The genes encoding the beta-subunits of ATP-synthases (ATPases) from Bacteroides fragilis DSM 2151, Cytophaga lytica DSM 2039 and 'Taxeobacter ocellatus' were cloned. The nucleotide sequences were determined completely for the genes of the first two organisms and to a major part for that of 'T. ocellatus'. The predicted amino acid sequences were compared with previously published amino acid sequences of beta-subunits. Two characteristic insertions were found in genes from organisms belonging to the so-called bacteroides-cytophaga-flavo-bacterium group. The remaining structure shows a high degree of sequence similarity within this group. These data support the conclusions drawn from comparative 16S rRNA sequence analyses that organisms in this phenotypically heterogeneous group are phylogenetically related. A phylogenetic tree was constructed based on a distance matrix of optimally aligned amino acid sequences of beta-subunits of ATPases of various eubacteria, chloroplasts and mitochondria. It is in good agreement with a tree derived from 16S rRNA sequence analyses. PMID- 2978297 TI - Dual roles for calcium ions in apical growth of Neurospora crassa. AB - We report initial attempts to define the role of Ca2+ in the polarized extension of Neurospora crassa. Growth of the organism was diminished in media containing less than 1 mM-Ca2+; extension was more severely impaired than biomass synthesis, resulting in the formation of stubby, bulbous hyphae, even of spherical cells. Reduced extension and abnormal morphology were correlated with the loss of surface-bound Ca2+, probably associated with the cell wall. Intracellular Ca2+ may be represented by material that fluoresces brightly in the presence of chlortetracycline. Punctate fluorescent bodies and diffuse fluorescence were both arrayed in a longitudinal gradient, maximum apically. Addition of the calcium ionophore A23187 induced dissipation of the fluorescence; concurrently, the hyphae lost as much as one half of their Ca2+ content. Extension continued almost unabated, but multiple branches quickly emerged from the apex. The observations suggest that a cytoplasmic Ca2+ gradient is not required for polarized extension, but may play a role in ensuring the dominance of the apex. PMID- 2978298 TI - [Systemic management of a patient with Marfan's syndrome during dental treatment]. PMID- 2978299 TI - [Progress in special emergency treatment]. PMID- 2978300 TI - AIDS: Clinic V and the role of GDPs. PMID- 2978301 TI - [Therapeutic trials in general practice]. PMID- 2978302 TI - [HTLV-I demonstrated in chronic progressive myelopathy]. PMID- 2978303 TI - [Motor disorders of the unoperated stomach]. PMID- 2978304 TI - [Effects of antihypertensive agents on intrarenal hemodynamics]. PMID- 2978305 TI - [Autoimmune hypoglycemia: the fault of pyritinol?]. AB - Initially reported in Japan, autoimmune hypoglycemia is related to the presence of insulin-binding antibodies, even in patients who have never been treated with insulin. The authors report a case of spontaneous autoimmune hypoglycemia in a French woman receiving pyritinol. The difference between insulin and C peptide radioimmunoassay levels prompted a search for insulin antibodies. In vitro studies confirmed their presence and showed that they were immunoglobulins G with two binding sites without species specificity. The outcome of autoimmune hypoglycemia is usually favourable, with a rapid decrease of insulin antibodies but steroid therapy is needed when serious clinical manifestations are present. The differential diagnosis with factitious hypoglycemia may be difficult. The reasons for the appearance of the insulin antibodies and the exact mechanisms of hypoglycemia remain hypothetical. However, drugs with a sulfhydril group, such as pyritinol, could play a causative role in this syndrome. PMID- 2978306 TI - [Prognostic factors in resuscitation]. AB - The outcome from intensive care is known to be influenced by such factors as age, previous health status, severity of the disease and diagnosis. In order to assess the influence of each individual factor, 3,687 patients from 38 French intensive care units were studied. For each patient were recorded: age, simplified acute physiological score (SAPS), previous health status, diagnosis, type of intensive care unit (medicine, scheduled or elective surgery) and immediate outcome. Each of these factors was found to influence the immediate survival rate. A multivariate analysis ranked the factors in the following order: SAPS, age, type of intensive care unit and previous health status. Diagnosis played a role in the prognosis since with a 10-15 points SAPS mortality was nil for drug overdose, 12 per cent for chronic obstructive pulmonary disease and 38 per cent for cardiogenic shock. However, a single diagnosis was made in only 37 per cent of the patients, as against 3 diagnoses in 17 per cent and 4 diagnoses or more in 7 per cent. When the type of intensive care unit was considered, the mean death rate was 20 per cent in medicine, 27 per cent in scheduled surgery and 5 per cent in elective surgery (P less than 0.001). Since this study showed a definite influence of each of the four factors on immediate survival, intensive care patients can be described and classified according to this system. However, it must be stressed that individual prognoses are extremely vague. PMID- 2978307 TI - [Complications and sequelae of thyroid surgery]. AB - A series of 408 operations performed on the thyroid gland is presented. The mortality rate was 0.25 per cent, and late recurrent nerve paralysis occurred in 2.2 per cent of the cases. This complication was independent of the disease and of the type of operation. Contrary to temporary recurrent nerve paralysis, which occurs with large goitres, it was observed in patients with small thyroid glands. Six patients had transient hypocalcaemia but none developed permanent hypoparathyroidism. The benign character of thyroid gland surgery is emphasized. The main morbidity, recurrent nerve paralysis, could be reduced by a strict surgical technique. Systematic parathyroid gland dissection and reimplantation account for the absence of late hypoparathyroidism. PMID- 2978308 TI - [Surgical treatment of leprous neuritis. The results in 114 operations]. AB - One hundred and fourteen surgical operations for neuritis were performed in 50 patients coming from areas where leprosy is endemic. The neurological signs included pain and/or sensorimotor deficit. The surgical procedure consisted of transposition and/or neurolysis. Pain subsided in 86 per cent of the cases, and the sensorimotor deficit was reduced in 78.9 per cent. The effects of surgery on pain always appeared on recovery from anaesthesia or on the day following the operation. The results were particularly good in young subjects and in patients with neurological signs of recent onset. Poor results were observed only in cases of old and painless neuritis with motor deficit, the latter being usually unchanged. Few studies have been published on large series of leprous neuritis patients treated surgically. Surgery in such cases must be associated with a medical treatment, failing which the patient is exposed to relapses. PMID- 2978309 TI - [Idiopathic calcifications of the central gray nuclei (Fahr's disease). Detection by magnetic resonance imaging]. PMID- 2978310 TI - [The "third testicle" in a cyclist: an ailment not to be ignored]. PMID- 2978311 TI - [Do nonsteroidal anti-inflammatory drugs have a protective effect against Campylobacter pylori?]. PMID- 2978312 TI - [Respiratory Mycoplasma pneumoniae infections in the acquired immunodeficiency syndrome]. PMID- 2978313 TI - [Use of amiodarone in pregnant women]. PMID- 2978315 TI - [Megalocytic interstitial nephritis associated with renal adenocarcinoma. A case]. PMID- 2978314 TI - [Aseptic purulent meningitis in Kawasaki syndrome in an adult]. PMID- 2978316 TI - [The value of cyclosporin in treating Wegener's disease]. PMID- 2978317 TI - [Diagnosis of atopy by a new test: Phadiatop]. PMID- 2978318 TI - [Etiological research on neutropenia in adults]. PMID- 2978319 TI - [Atraumatic, as far as possible, neonatal resuscitation]. PMID- 2978320 TI - [Transesophageal cardiac stimulation. Its value and limits]. PMID- 2978321 TI - [Drug-induced thrombopenias and neutropenias. An in vitro study by the indirect immunofluorescence test]. AB - The sera from 38 patients with suspected drug-induced thrombocytopenia (22) or neutropenia (16) were tested with the indirect immunofluorescence test on platelets or granulocytes for the presence of drug-dependent antibodies. Three drug-induced antibodies with reactivity against platelets and 5 with reactivity against granulocytes were detected. In 3 sera antibodies were found which reacted already with target cells without adding the drug to the test system. These data show that drug-induced blood dyscrasias often have an immunological cause and that in vitro tests can be helpful in detecting the responsible drug. Different mechanisms can be involved. In many sera circulating antibodies were not found, but an immunological mechanism is likely to be involved in some of these cytopenias: the antibody could be entirely absorbed by the target cells, a metabolite of the drug could be the immunogen and finally the test may not be sensitive enough. PMID- 2978322 TI - [Sarcomatous recurrence of leiomyoma of the inferior vena cava]. AB - In a 63-year old woman, a surgically removed leiomyoma of the inferior vena cava recurred, 4 years later, as a leiomyosarcoma. These two smooth muscle tumours, one benign the other malignant, seldom involve the inferior vena cava. Both produce few clinical symptoms and therefore are belatedly diagnosed, except for suprarenal or retrohepatic lesions which rapidly exhibit signs of compression. Computerized tomography, angioscans and cavography are necessary to make a provisional diagnosis and determine the operative strategy. The differential diagnosis between leiomyoma and leiomyosarcoma rests on cellular or nuclear atypias, mitotic index and sometimes cytological and staining details showing the myofibril. These tumours develop slowly in the renal segment. Treatment is primarily surgical. The case reported here raises the problem of the leiomyoma leiomyosarcoma sequence: are we confronted with degeneration, or is there such a thing as a smooth muscle fibre disease? PMID- 2978323 TI - [Systemic toxicity of paraphenylenediamine. 4 cases]. AB - Paraphenylenediamine (PPD) poisoning has been known for many years, but few studies have been devoted to the subject. We report 4 cases, seen in the nephrology department, concerning women aged from 18 to 35 years who had tried to commit suicide by drinking varying amounts of that hair dye. The initial symptom was acute asphyxia which required emergency tracheotomy in 3 cases. Thereafter, the most important visceral damage was acute renal failure, usually with oliguria or anuria, for which haemodialysis was performed in 2 cases. In these patients treated at an early stage the mid- and long-term prognosis was satisfactory. Rhabdomyolysis is the principal mechanism underlying PPD systemic toxicity; it is, in particular, responsible for the renal failure observed. The selling of PPD hair dye should be restricted by law. PMID- 2978324 TI - [Metoprolol, a new effective antimigraine agent]. AB - The effectiveness of metoprolol as a prophylactic treatment of migraine was investigated in double-blind comparative trials involving more than 500 patients. In two trials using a slow-release preparation and a dose of 200 mg per day, metoprolol proved significantly better than a placebo and as effective as propranolol 160 mg/day or flunarizine 10 mg/day. At the dose of 100 mg per day metoprolol was significantly more effective than clonidine and clomipramine and not different from a placebo, propranolol 80 mg/day or pizotifen 1.5 mg/day. It is concluded that metoprolol is useful as prophylactic treatment of migraine and ranks among the main drugs used for the same purpose, such as propranolol and pizotifen. PMID- 2978326 TI - [D8-D9 dorsal herniated disk revealed by spinal cord compression]. PMID- 2978325 TI - [Congenital tracheal stenosis due to complete circular rings. Tracheoplasty using autologous pericardium]. AB - The case of a 9-month old infant with congenital tracheal stenosis due to complete tracheal rings, associated with an aberrant left pulmonary artery is reported. Surgery was carried out under extracorporeal circulation to ensure adequate oxygen supply. Extubation, thus made possible, enabled the lesion to located accurately by bronchoscopy. Tracheoplasty by means of an autologus pericardial patch was performed. The quality of the repair was checked by a second bronchoscopy. The long-term result is satisfactory. PMID- 2978328 TI - [Adrenal cortex function in patients with the acquired immunodeficiency syndrome]. PMID- 2978327 TI - [Zidovudine efficacy and tolerance in 32 patients with cerebral toxoplasmosis in the acquired immunodeficiency syndrome]. PMID- 2978329 TI - [Sleep polygraph study of African subjects in Niger]. PMID- 2978330 TI - [Joint periostitis of the lower limb disclosing a fistulated and infected aortic prosthesis]. PMID- 2978331 TI - [Streptococcal group A fulminant septicemia at the end of pregnancy]. PMID- 2978332 TI - [Complete wrapping of the liver with resorbable mesh for hepatic rupture with persistent hemorrhage]. PMID- 2978333 TI - [An unusual cause of hypertension in an elderly subject and its consequences]. PMID- 2978334 TI - [Calculus, bile, statistics and the surgeon]. PMID- 2978335 TI - [Serratia marcescens nosocomial septicemia and pseudobacteremia]. PMID- 2978336 TI - The stroke patient. PMID- 2978337 TI - [Biomechanical considerations concerning the behavior of the sustaining periodontium with respect to the site at which the mastication force is exercised]. PMID- 2978338 TI - [Electron optical aspects of the constitution and organization of the bacterial dental plate (Part I)]. PMID- 2978339 TI - [The antimicrobial action of a complex metronidazole paste on the microbial flora in infected root canals]. PMID- 2978340 TI - [Primary malignant melanomas of the buccal cavity]. PMID- 2978341 TI - [Malignant tumors of the buccomaxillofacial nervous structures]. PMID- 2978342 TI - [The hemostatic effect of an adhesive apiphytotherapeutic preparation--Propovit M.S.K. experimental study]. PMID- 2978343 TI - [Mathematical and algorithm methods for interpretation of the Teleroentgen cephalometric images with a view to establishing the facial developmental prognosis of children during their growth]. PMID- 2978344 TI - [Evaluation of the efficiency of the cariopreventive chewing gum produced in Romania]. PMID- 2978345 TI - [Binder's syndrome. Comments with reference to a clinical case]. PMID- 2978346 TI - [Contributions to the elaboration of anamnestic indexes with epidemiologic character, in view of detecting the frequency of the craniomandibular painful dysfunctional syndrome, associated or not with recurring migraine (Note I)]. PMID- 2978347 TI - [The use of resilient materials for improving the functionality of mobile prostheses]. PMID- 2978348 TI - [A clinical and therapeutic study of temporo-mandibular arthroses]. PMID- 2978349 TI - [Implications of oral respiration in dento-facial orthopaedics]. PMID- 2978350 TI - [Epidemiology of stomatological affections in preschool children]. PMID- 2978351 TI - [The study of monosaccharides and of basic amino acids from the structure of glycoproteins isolated from bacterial dental plaque]. PMID- 2978352 TI - [Particularities of pulp pathology in subjects with capillary fragility]. PMID- 2978353 TI - [The painful T.M.J. syndrome in the totally edentate with total prosthesis]. PMID- 2978354 TI - [Surgery in complex pathology of the ascending aorta: replacement of the ascending aorta, aortic valve and reimplantation of the coronary arteries in 6 patients]. PMID- 2978355 TI - [Fanconi's anemia: effect of treatment with a vitamin complex and with nicotinamide]. PMID- 2978356 TI - [Modulation of Ca2+-ATPase localization in early dentinogenesis of rat incisor]. PMID- 2978358 TI - [Improving regional health education propaganda]. PMID- 2978357 TI - The enigma of the acquired immunodeficiency syndrome. PMID- 2978359 TI - [The need of specialized services for patients receiving ambulatory care]. PMID- 2978360 TI - [Medico-demographic characteristics of young large families in the Kazakh SSR]. PMID- 2978361 TI - [Medico-social characteristics of gynecologic patients in need of rehabilitation treatment]. PMID- 2978362 TI - "The dental implications of HIV infection". A report on the content of a conference held at Sheffield, U.K. 13th & 14th July, 1988. PMID- 2978363 TI - [Osteomas of the paranasal sinuses: review of the literature and personal cases]. AB - The sinusal osteomas: a review of literature and personal cases. The osteoma is a osteogenetic benign tumour formed by a fully developed bone, characterized by a slow growth, which, as far as the maxillo-facial region is concerned, shows to be frequently located in the paranasal sinuses. The osteoma originates from one of the sinus walls and spreads into the sinus cavity itself producing symptoms related to a lesion space occupying. In the present report, taking into consideration 4 personal cases, some literature datas are examined concerning the epidemiology, clinical behaviour and especially the therapeutic implications to be adopted by the surgeon in the presence of sinusal osteoma. PMID- 2978364 TI - [Water loading diuresis test in albino rats. Critical study of approximately 12,000 tests. 3. Loading tests using several mineral waters bottled under various experimental conditions]. AB - The authors studied a series of more than 12,000 complex diuresis exams after water loading, in the albino rat, which permitted them to draw the following conclusions: 1). The temperature of water definitively and, probably the environmental temperature, influence the diuretic response. 2). The most favourable water loading preparations are not fasting and solid fasting during the 24 hrs. prior to loading. 3). Pretreatment with diuretic water 15 days prior to loading, favours the diuretic response to the same or the any other water: activated kidney. 4). The addition of CO2 to water, by itself, does not seem to favour diuresis. 5). Aging or conservation in bottles, always reduces, more or less sensitively, the diuretic properties of the water itself, especially during the first 2-3 months. This confirms that the diuretic properties do not only and exclusively depend on the chemical or physical composition or on the structure, but on everything together, that is modified after the moment of it's natural emergence. This loss of activity, in any case, is favourable to the indiscriminate use of these waters as both dietetic and drinking water. 6). It is probable that the different pH's of water influence it's diuretic activity. 7). Even if the more active waters, in this study, seem to be bicarbonate, sulfate bicarbonate, or bicarbonate-sulfate-alkaline-terrose, this can be attributed to the prevalence of these classes of water in this study. Instead, what emerges with certainty, from the point of view of the molar concentration, is that while there are which are minimally mineralized and oligominerals with scarce or no diuretic activity, there are mediomineral and hypotonic mineral waters with up to 100 mmol/liter, and with conspicuous diuretic activity, even after more or less long periods of conservation in bottles. 8). For all of the above-mentioned reasons, we feel that a useful orientation can also be gained for the use of the discussed water for human needs. PMID- 2978365 TI - [The hypothalamic-hypophyseal neurosecretory system of the Alosa fallax nilotica. A histological and ultrastructural study]. AB - We studied the morphological, histochemical and ultrastructural aspects of the hypothalamic-pituitary system in the Alosa fallax nilotica. The secretory activity of the preoptic nucleus and of the lateral tuberal nucleus was also investigated with regard to the gonadal growth and development during the reproductive sexual cycle and the changing migrational environmental conditions (anadromous and catadromous phases). The hypothalamic-pituitary system is formed by: centers of neurosecretion synthesis (parvo and magnocellular preoptic nucleus and lateral tuberal nucleus), a conduit system (preoptic infundibular neurohypophyseal tract) and a collecting center (neurohypophysis intermediate lobe). The neurosecretory cells show two distinct types of secretory granules with different ultrastructural features probably representing the morphological expression of two distinct hormones. These cells show also characterizations of cytoplasmic areas related to peculiar cyclic functional stages concerned with the reproductive sexual cycle. We observed significant quantitative and qualitative changes of the neurons both in the parvo- and magnocellular preoptic nucleus and in the lateral tuberal nucleus during anadromous migration and during spawning and fertilization. Almost insignificant changes of the neurosecretory material were observed in the preoptic-pituitary tract and in the intermediate neurohypophyseal lobe. The results show that the hypothalamic-pituitary neurosecretory system play an important role in the osmoregulation (magnocellular preoptic nucleus) and on the reproductive cycle (lateral tuberal nucleus). The system probably also controls some other metabolic activities (utilization of the stored material) linked to peculiar environmental and feeding conditions. PMID- 2978366 TI - [Acute cardiotoxicity of tricyclic compounds: remarks on a clinical case]. AB - Tricyclic compounds have complex effects on the cardiovascular system, causing dangerous arrhythmias even in patients free of cardiac disease, especially after overdose. We report on a case of cardiac arrhythmia during low dosage therapy of post-herpetic neuralgia with Carbamazepine and Amitriptyline in a patient with chronic renal rejection and cirrhosis. Pharmacologic properties and side effects of these drugs are examined from the current literature. Both drugs prolong AV conduction, consequently You can observe AV blocks of various degrees of His bundle branch blocks also during low-dosage therapy in patients with hepatic failure. It seems to be judicious to provide to ECG monitoring during the complete period of therapy. PMID- 2978367 TI - [The contribution of the Doppler method in the diagnosis of Raynaud's syndrome]. AB - According to this method, during the period of the crisis, the blood flow graph of the digital arteries changes in the opposite direction when this is compared with the blood flow graph during the period of calmness which is of course the normal state. PMID- 2978368 TI - [Diagnosis of depression in chronic alcoholism: methodological aspects]. AB - The incidence of depression ranges between 3% and 98% in chronic alcoholism. This discrepancy has been attributed to the lack in univocal diagnostic criteria and to poor attention to the moment when the observation takes place. In patients with chronic alcoholism, accurate clinical examinations are required to make a diagnosis of depression; furthermore, the presence of continuous alcohol abuse, or the condition of initial of prolonged abstinence must be considered. Using these criteria the diagnosis of depression in chronic alcoholism will probably become more reliable and will be possible to carry out a more appropriate therapy. PMID- 2978369 TI - First clinical experience with a new TCR/CD3-monoclonal antibody (BMA 031) in kidney transplant patients. PMID- 2978370 TI - Doxazosin, a case history. AB - Demonstration that the antihypertensive activity of prazosin resulted from blockade of postjunctional alpha 1-adrenoceptors stimulated interest in identifying the structural basis for this novel pharmacological profile. SARs suggested that 2,4-diamino-6,7-dimethoxyquinazoline was the pharmacophore for alpha 1-adrenoceptor recognition and that receptor occupancy was similar to noradrenaline. This model stressed the importance of charge-reinforced hydrogen bonding rather than particular roles played by individual nitrogen atoms. Modification of the quinazoline 2-substituent influenced alpha 1-adrenoceptor affinity and antihypertensive activity, particularly duration of action. These studies led to a novel series of quinazoline-benzodioxanoyl derivatives from which doxazosin was selected for detailed evaluation. Doxazosin is a potent, selective alpha 1-adrenoceptor antagonist which produces long-lasting antihypertensive effects in laboratory animals, with little effect on heart rate. In man, the plasma half-life of doxazosin is approximately 10 hr., consistent with clinical evidence that once-daily administration provides 24 hr. blood pressure control. PMID- 2978371 TI - Characteristics and uses of natural killer cells. PMID- 2978372 TI - Differential expression of the leucocyte-common antigen family. PMID- 2978373 TI - Human naive and memory T cells: reinterpretation of helper-inducer and suppressor inducer subsets. PMID- 2978374 TI - An in-vitro model for tumor immunotherapy with antibody heteroconjugates. PMID- 2978375 TI - Cell surface heparan sulphate implicated in haemopoietic growth factor signalling. PMID- 2978376 TI - B-cell transplants to immunodeficient xid neonates do not imprint their helper T cells. AB - A great variety of Igh-restricted immunoregulatory phenomena have been described in responses to both haptens and carriers. The investigation of Igh-dependent T cell functions is of considerable interest because it relates to issues such as the interdependence of the T- and B-cell networks, isotope- and idiotype-specific help, the role of Igh-linked products in the generation of the helper and cytolytic T-cell repertoires and Igh-linked effects on cell interactions among helper and suppressor cells. Helper T cells have been shown to be critically susceptible to B cell-mediated education and it is generally assumed that T cells become imprinted by B-cell idiotypes during development. It occurred to us that xid immunodeficient neonates, which lack T15+ B cells and T15 antibodies, might provide a suitable host to investigate the B-cell dependent imprinting of T cells providing help in T15+ anti-PC responses. We designed our experiments to test whether or not T15 deficient xid mice could be used as recipients of normal B cells capable of imprinting the host's helper cells. The results of our experiments demonstrate that unirradiated xid neonates are readily engrafted with normal B cells from both neonatal and adult donors. The transplanted B cells reconstitute anti-PC and anti TNP-FICOLL PFC responses in the xid hosts. However, the early engraftment of T15+ B cells in xid mice did not cause a detectable imprinting of their helper T cells, i.e., we could not detect any biases in the preference of carrier-/primed cells from reconstituted mice for primary or secondary T15+ or T15- B cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978378 TI - Estimation of premorbid intelligence in organic conditions. AB - The validity of premorbid IQ estimates provided by the National Adult Reading Test (NART) and Vocabulary sub-test of the Wechsler Adult Intelligence Scale were evaluated, by comparison with matched, healthy control subjects, in Korsakoff psychosis, alcoholic dementia, dementia Alzheimer type (DAT), multi-infarct dementia (MID), Huntington's disease, and closed head injury (CHI). There was no significant difference in NART performance between control subjects and the alcoholic dementia, DAT, MID, and CHI groups. Although there appeared to be a decline in NART performance in the Korsakoff and Huntington's groups, it did provide a significantly higher IQ estimate than the Vocabulary sub-test. All clinical groups, with the exception of the CHI group, performed at a significantly lower level than the control group on the Vocabulary sub-test. PMID- 2978379 TI - Mianserin and mania. PMID- 2978377 TI - Artificial dimers of native actin: preparation and properties in biological functions. AB - With the aid of tartryl-bis-epsilon-aminocaprylazide artificial dimers were produced from F actin from rabbit striated muscle. These derivatives will not polymerize by themselves but are able to copolymerize fully with native G actin. By modification of a single side chain per dimer, this copolymerization was completely inhibited. The dimers are able to activate subfragment 1 ATPase of myosin and bind to DNase I with inactivation of the enzyme in the same manner as native G actin. Within the dimer, one ADP is immobilized and will exchange against ATP extremely slowly. The dimers do not bind to the mushroom toxin phalloidin. PMID- 2978380 TI - A study of mortality experiences in a mental-handicap hospital. AB - The study examined the mortality experiences of a population of mentally handicapped in-patients between 1983 and 1987. Demographic data and factors related to terminal illness and cause of death were considered. The findings suggested a continued trend towards increased longevity and indicated a number of groups at risk of early death. PMID- 2978381 TI - [Surgical treatment of acne scarring: non-linear scar revision]. PMID- 2978382 TI - [Liquid nitrogen therapy of acne vulgaris]. PMID- 2978383 TI - [Down syndrome in Chinese-radiographic assessment in neonatal period]. PMID- 2978384 TI - Actuality in the treatment of multivessel unstable angina pectoris. PMID- 2978385 TI - In vitro interactions of carumonam in combination with coumermycin, teicoplanin, flucloxacillin and vancomycin. PMID- 2978386 TI - Interpretive criteria for the agar diffusion susceptibility test with teicoplanin using the ICS-and DIN-methods, respectively. PMID- 2978387 TI - T lymphocyte subpopulation in early and late schistosomal hepatosplenomegaly. PMID- 2978388 TI - Therapy of Clostridium difficile in pseudomembranous and antibiotic-associated colitis. PMID- 2978389 TI - Myonecrosis of the abdominal wall, complicating radical hysterectomy. PMID- 2978390 TI - Vaccination campaign against hepatitis B in Italy. PMID- 2978391 TI - New vaccines against hepatitis B. PMID- 2978392 TI - [Ultrastructural changes in hemopoietic microenvironment and hemopoietic cells in bone marrow of aplastic anemia]. PMID- 2978393 TI - [Hereditary angioedema with chronic glomerulonephritis]. PMID- 2978395 TI - [Effects of retinoids on tumor specific immune response in mice]. PMID- 2978394 TI - [Studies on antimicrotubule and antitumor activities of synthetic PD derivatives]. PMID- 2978396 TI - [Construction of plasmids containing different human globin gene fragments]. PMID- 2978397 TI - [Study on biological characteristics of spontaneous diabetic KK mice]. PMID- 2978398 TI - [Experimental study of atherosclerosis. II. The effect of 8501 on the balance between thromboxane A2 and prostacyclin in preventing experimental atherosclerosis of rabbits]. PMID- 2978399 TI - [Immunoregulatory effects of ascites of tumor-bearing mice]. PMID- 2978400 TI - [DNA fragments labeled with biotin]. PMID- 2978401 TI - [Effect of dermatan sulphate on plasminogen activator activity in perfused rat hindlegs]. PMID- 2978402 TI - [Inhibitive effect of anisodamine on increase of vascular permeability after injection of Escherichia coli endotoxin in mice]. PMID- 2978403 TI - [Effect of racemic, (+),(-) gossypol on rat testis, epididymis, liver and kidney]. PMID- 2978404 TI - [Detection of anti-DNA antibody by horseradish peroxidase ELISA]. PMID- 2978405 TI - [Neuronal tracing with choleragen subunit B--a monoclonal immunocytochemical method]. PMID- 2978406 TI - [Ms-ELISA for detection of serum antibody level in leprosy patient--establishment of Ms-ELISA]. PMID- 2978407 TI - [Observation on plasma serotonin (5-HT) and platelet membrane fluidity in AMI patients]. PMID- 2978408 TI - [Post-treatment evaluation of fillings with Cermet cement (Ketac-Silver)]. PMID- 2978409 TI - [First clinical results of retrograde root canal filling using a Cermet cement]. PMID- 2978410 TI - [Back problems and bad postural habits]. PMID- 2978411 TI - [Psychological training in dentistry. 2]. PMID- 2978412 TI - [Dental health measures to psychopathic patients. I. The condition of oral hygiene of psychiatric inpatients]. PMID- 2978413 TI - Electrocardiographic findings in healthy adults residing at high altitudes in the Himalayas. PMID- 2978414 TI - Beta-endorphin levels after intrathecal infusion of iodinated contrast media. AB - Modifications in beta-endorphin levels in cerebrospinal fluid have been described following lumbar puncture and metrizamide injection. Cerebrospinal fluid (CSF) samples were obtained from 19 patients before and after lumbar myelography. Two radioimmunoassays were used. One was a commercial kit; and the other one (developed in our laboratory) used a chromatographic removal from beta lipotrophin. No definite variation of beta-endorphin was observed after myelography, using either the commercial kit or a more sophisticated procedure. Some controls were prepared by adding metrizamide or Iopamidol in vitro to CSF samples in order to evaluate a non specific effect of these contrast media. The results obtained with these controls suggest that the discrepancy of results may be explained simply by assay artifacts due to drug interferences when using the commercial methods. PMID- 2978415 TI - An analysis of the effect of HLA-DP in the mixed lymphocyte reaction. AB - It has recently been reported that HLA-DP antigens may play an important role in the development of graft-versus-host disease (GVHD) following transplantations of haploidentical bone marrow as a treatment for haematological malignancies. Mixed lymphocyte culture (MLC) is routinely performed prior to bone marrow transplantation to assess the suitability of the donor, and we have therefore examined the role of HLA-DP in this test. One-way MLC chequerboard experiments were performed between 17 HLA-Dw3 homozygous typing cells (HTC) with a range of HLA-DP antigens represented, including HLA-DPw1, w2, w3, w4 and CP63. The experiments were performed on multiple occasions and each time a highly significant difference (P = less than 0.001) was observed between the Relative Responses (RR) in the HLA-DP matched responder/stimulator pairs and the HLA-DP mismatched pairs. There was, however, considerable overlap in these results with ranges in the HLA-DP-matched group RRs of 0-17%, and 0-62% in the mismatched group. Only 3.1% of the HLA-DP-matched grou had a RR greater than 5%, while 48% of the HLA-DP mismatched group had a RR greater than 5%. From these results it was calculated that a positive response (greater than 5%) has a 96% chance of being due to an HLA-DP disparity of one or two antigens. Conversely, with a negative MLC the chance of their being no HLA-DP antigen disparity was only 65%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978416 TI - Dentists' professional and ethical responsibilities for HIV-positive patients and patients with AIDS. PMID- 2978417 TI - Awareness and acceptance of hepatitis B vaccine by Irish dental practitioners. PMID- 2978418 TI - Graded spinal cord injuries produced in rabbits with non-invasive microwave hyperthermia. AB - The use of non-invasive microwave energy to produce spinal cord injuries with intraspinal hyperthermia was studied in experimental animals. Lesions were produced with external beam microwave irradiation at 915 MHz in rabbits, using intraspinal temperature levels from 40 to 43 degrees C., and periods of heating ranging from 15 to 60 minutes. The parameters which determined thermal dose were the degree of temperature elevation in the spinal cord relative to the body core and the duration of that elevation. Thermal dose-response relationships were established by monitoring intraspinal temperatures during heating using an epidural thermistor probe at T8. Animals were examined 48 hours after lesion production and assigned a neurological grade. Injuries were grouped clinically according to their degree of relative functional severity as minimal, mild, moderate, or severe. Evaluation of spinal cord integrity was carried out by recording cortical somatosensory evoked responses (SER) following sciatic nerve stimulation. Increased SER latencies were first observed after heating the spinal cord to 41 degrees for 60 minutes. Impulse transmission was absent after heating to 42 degrees for 30 minutes, a thermal dose which produced complete paraplegia. Morphologically, lesion size and configuration were directly related to the thermal dose used in their production. Low thermal doses produced white matter edema limited to the posterior columns, while larger doses resulted in demyelination, retrograde neuronal changes, and infarction of the dorsal half of the cord. High thermal doses also produced foci of hemorrhage in the gray and white matter of the dorsal cord. These studies suggest that reproducible spinal cord injuries with predictable levels of neurological severity can be produced by noninvasive microwave heating.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978419 TI - Papillon Lefevre syndrome--a case report. PMID- 2978420 TI - Prescribing requirements of the elderly mentally handicapped: future demands on primary health care teams. AB - The medication prescribed for 269 mentally handicapped hospital residents aged 60 years and over on 1 January 1987, including five with Down's syndrome, and for 31 residents aged 50 years or over with Down's syndrome was analysed.Fifty of the 269 elderly residents were receiving no medication but the mean number of prescriptions for the remaining 219 was four. Of the total sample of 269 residents, 32% were taking drugs for psychiatric and psychological disorders; 11% regular antiepileptic drugs; 62% long-term drugs for other problems (diuretics and laxatives were the most common); 29% had long-term prescriptions for topical preparations, enemata or suppositories; 23% were on shortterm prescriptions for other problems; and 17% had short-term prescriptions for topical preparations, enemata or suppositories. For the 31 Down's syndrome residents the commonest prescriptions were for skin preparations.Particular care is needed in making a diagnosis and monitoring the effects of treatment on mentally handicapped patients because a complete history and full cooperation on examination may not be forthcoming. These findings have implications for primary health care teams who will be responsible for the mentally handicapped when hospitals close and the residents live in the community. PMID- 2978421 TI - Carotid artery Dacron patch graft angioplasty: a ten-year experience. AB - Of 394 carotid endarterectomies performed during a 10-year period, 36 (9%) included Dacron patch graft angioplasties. Patch graft angioplasties were used selectively in patients because they had undergone previous carotid endarterectomy or had small-caliber vessels. There were no postoperative infections, occlusions, emboli, stroke, or pseudoaneurysms. In addition, no clinically detectable recurrent stenoses developed in this high-risk group of patients. Twenty-four of the 27 patients available for follow-up were screened for asymptomatic stenoses with spectral analysis and B-mode imaging; there was no sign of hemodynamically significant recurrent stenosis in this group. These encouraging results support the continued selective use of Dacron patch graft angioplasty in carotid surgery. PMID- 2978422 TI - Development of blood flow velocimeter for ocular vessels. AB - Clinical evidence suggests that blood flow in the retinal vessels varies in progressing diabetic retinopathy, coronary arteriole disease, hypertension and some other clinical situations. The changes of the flow in the blood vessels may serve as a monitor of the disease and as a follow up device for treatment success. Despite their importance direct blood flow velocity measurements are not easily obtainable. Recent advances in the development of a laser Doppler anemometry for the determination of in-vivo retinal blood velocity enables to obtain, with a noninvasive technique, information relevant to the flow of blood in the human retina. The described anemometry system is designed to be integrated, self-aligned and to operate with backscattered light. The influence of the important design parameters as well as focusing of the control volume on the retinal arteries is analyzed. Due to multiscattering of light from the blood cells present simultaneously in the control volume, the electronic signal obtained is very difficult to process and sophisticated signal processing routines are still being developed. At present measurements are used to obtain results with blood flowing in glass tubes of an inside diameter of 50 to 2000 microns. Preliminary experimental and theoretical simulations are currently being performed to validate the principle of the proposed data processing technique. Initial measurements have already produced relative high quality (and thus processable) signals, indicating good prospects for a practical ophthalmic system operation. PMID- 2978424 TI - [Native myosins of the masticatory muscles of the cat using non-dissociative gel electrophoresis]. PMID- 2978423 TI - Calmodulin-activated calcium ATPase in bipolar illness. AB - Calmodulin-activated calcium ATPase is a transport enzyme which establishes the normal level of intracellular ionized calcium in most cells. We have determined values for three parameters of this enzyme: E-t, the concentration in the membrane; Vmax, the maximal velocity, and Ka, the binding affinity for calmodulin. We assayed these parameters in erythrocyte membranes from lithium carbonate-treated bipolar subjects and from normal controls. Bipolar subjects have significantly increased levels of E-t compared with normal controls. PMID- 2978426 TI - [Septicemia--an interdisciplinary clinical problem]. PMID- 2978425 TI - [The height of the palatal vault in infants with Down's syndrome]. PMID- 2978427 TI - [Principles of the diagnosis and organization of medical services for patients with septicemia]. PMID- 2978428 TI - [Sensitivity of Staphylococcus aureus strains to 2d and 3d generation cephalosporins]. PMID- 2978430 TI - The management of intermittent claudication. PMID- 2978429 TI - Post-herpetic abdominal wall herniation. PMID- 2978431 TI - Topical and oral tetracycline in the treatment of acne vulgaris. PMID- 2978432 TI - Lymphomatoid granulomatosis--evidence of a clonal T-cell origin and an association with lethal midline granuloma. AB - Lymphomatoid granulomatosis and lethal midline granuloma are both characterized histologically by atypical pleomorphic angiocentric infiltrates. Whether these conditions are malignant lymphoproliferative disorders remains controversial. Here we report the results of studies carried out in a patient with coeliac disease, who developed recurrent self-healing subcutaneous nodules with the histological changes of lymphomatoid granulomatosis and an invasive nasal tumour with the histological features of lethal midline granuloma. The patient subsequently also developed an erythrophagocytic syndrome. Immunocytochemical labelling of both cutaneous and nasal lesions demonstrated a predominant population of T cells with a CD4-negative CD8-positive phenotype. Analysis of DNA from cutaneous tissue revealed a discrete rearrangement of the beta and gamma T cell receptor genes. These findings suggest that lymphomatoid granulomatosis is a clonal T-cell lymphoproliferative disorder and its association with lethal midline granuloma indicates that both conditions may have a common histogenesis. PMID- 2978434 TI - [Medical considerations in geriatric dentistry]. PMID- 2978433 TI - Skeletal alterations in Down's syndrome: bibliographic review and case study contribution. PMID- 2978435 TI - [Current aspects of hemostasis]. PMID- 2978436 TI - [Infantile epistaxis: a local disease or a symptom of blood dyscrasia?]. PMID- 2978437 TI - [The limits of transtympanic injections in treating serous otitis with a closed tympanum]. PMID- 2978438 TI - [Our clinical management in treating seromucous otitis]. PMID- 2978439 TI - [Conductive hypoacusis and injuries of the middle ear]. PMID- 2978440 TI - [Etiologic aspects of chronic unilateral perceptive deafness]. PMID- 2978441 TI - [Acupuncture analgesia in pathology and the treatment of ENT injuries]. PMID- 2978442 TI - [Hypoacusis from simple ear injuries or associated with craniocerebral injuries]. PMID- 2978443 TI - [The role of otorhinolaryngologists in treating severe craniocervicofacial injuries]. PMID- 2978444 TI - [A tracheobronchial foreign body tolerated for 41 days]. PMID- 2978445 TI - [Malignant degeneration of laryngotracheal papillomas]. PMID- 2978446 TI - [Treatment possibilities in rhinopharyngolaryngo-tracheobronchial scleroma]. PMID- 2978447 TI - [Rechanneling of the nasal fossae with alloplastic material in ozena]. PMID- 2978448 TI - [Leucotrofina in the treatment of protracted suppurative otitis media in infants (preliminary note)]. PMID- 2978449 TI - [Pension Fund Administration. Disabled who received a pension prematurely]. PMID- 2978450 TI - [Inhibition of hypersensitivity reactions by oleanolic acid]. PMID- 2978451 TI - Activation of plasmin in mastitic milk. PMID- 2978452 TI - Immunofluorescent and immunoelectron microscopic localization of protein antigens in red cells infected with the human malaria Plasmodium falciparum. AB - Erythrocytes infected with the human malaria Plasmodium falciparum produce elevations of the surface membrane of the red cell called knobs. Through the use of transmission electron microscopy and a post-embedding protein A-immunogold technique, it was possible to show changes in the distribution of band 3, glycophorin A and spectrin in the region of the knob. These proteins appeared to be aggregated or condensed in the area of the knob, whereas the remainder of the red cell surface showed no such dense clusters; haemoglobin and the histidine rich protein of P. lophurae could not be localized to the knobby protuberances. It was not possible to detect any changes in protein distribution using the light microscope and indirect immunofluorescence. PMID- 2978453 TI - [Impairments of the locomotor system for the purposes of the new pension for disability (law 222 of 12-6-84). Diagnostic methodology and evaluation guidelines]. PMID- 2978454 TI - T-cell imbalances and NK activity in varicella-zoster virus infections. AB - Samples of peripheral blood lymphocytes (PBMC) were serially obtained from 30 patients with herpes zoster (HZ) and 10 patients with chickenpox (CP). Cells were assayed for NK-cell function and for the expression of surface membrane antigens which identify T-cell and NK-cell subsets. During the acute phase of disease (less than 7 days from onset), PBMC from patients with HZ had low proportions of T-helper (CD 4+) cells and a large number of T-suppressor (CD 8+) cells, resulting in a low T-helper/T-suppressor ratio. There was an increased percentage of nonspecific suppressor cells (GD 8+-CD 11+ cells) and increased expression of HLA-DR determinants on both CD 8+ and CD 4+ cells. The NK activity was depressed with no concomitant decrease in NK cells (CD 16+ or Leu 7+ cells). In the early convalescing phase of disease (8-14 days), there was a significant increase in CD 16+ cells and increased expression of HLA-DR on these cells, correlating with increased NK activity. In the late recovery phase (greater than 14 days), NK activity and levels of T-cell subpopulations were normal with the exception of increased CD 4+ cells and, consequently, of the helper/suppressor ratios. In the acute phase of CP (less than 7 days), the T-cell imbalances were similar to those encountered with HZ patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978455 TI - Induction of suppression of delayed type hypersensitivity to herpes simplex virus by epidermal cells exposed to UV-irradiated urocanic acid in vivo. AB - Urocanic acid (UCA), the putative photoreceptor for ultraviolet radiation (UV) induced suppression, undergoes a UV-dependent trans to cis isomerisation. Epidermal cells from mice painted with UCA, containing a known proportion of the cis-isomer, generate suppression of the delayed type hypersensitivity response to herpes simplex virus type 1 (HSV-1) when transferred to naive syngeneic recipients at the same time and site as infection with HSV-1. One T suppressor cell subset, of phenotype (Thy1+, L3T4+, Ly2-), is induced by the cis-UCA modified epidermal cell transfer. Flow cytometric analysis of the epidermal cells from skin treated with UV or cis-UCA indicates an overall reduction from normal in the number of cells expressing MHC Class II antigens, but no alteration in the number expressing I-J antigens. PMID- 2978456 TI - Diminished interferon gamma production may be the earliest indicator of infection with the human immunodeficiency virus. AB - The degree of clinical severity in human immunodeficiency virus infected patients, ranging from asymptomatic seropositive subjects to acquired immune deficiency syndrome, as well as in individuals at risk was assessed in relation to: (1) T-cell subset balance and expression of markers of T-cell activation; (2) natural killer activity; and (3) interferon gamma production. A decrease in the CD4/CD8 (helper/suppressor) ratio and an increase in the percentage of CD8+ (suppressor/cytotoxic) cells coexpressing markers of activation (HLA-DR or CD25) were closely correlated with the clinical severity of the human immunodeficiency virus infection. Natural killer activity was significantly impaired in patients with acquired immune deficiency syndrome and acquired immune deficiency syndrome related complex but normal in asymptomatic seropositive individuals and subjects at risk. Interferon gamma production, either in response to mitogens or the antigens from infectious agents commonly affecting human immunodeficiency virus positive individuals, was decreased in patients with acquired immune deficiency syndrome or acquired immune deficiency syndrome-related complex, with lesser involvement in human immunodeficiency virus-seropositive subjects or individuals at risk. Four of the six persons in the last group seroconverted during the ten months subsequent to evaluation of their immune status. Since production of interferon gamma was diminished in these patients while other assays of immunity were normal, measurement of this lymphokine may be a useful determinant of infection with the human immunodeficiency virus. PMID- 2978457 TI - Specificity and function of T cells bearing gamma delta receptors. PMID- 2978458 TI - Selenocysteine, a highly specific component of certain enzymes, is incorporated by a UGA-directed co-translational mechanism. AB - The opal termination codon UGA is used in both prokaryotic and eukaryotic species to direct the specific insertion of selenocysteine into certain selenium dependent enzymes. So far a formate dehydrogenase (hydrogenase-linked) of Escherichia coli and glutathione peroxidases of murine, human and rat origin have been identified as enzymes containing selenocysteine residues encoded by UGA. A novel seryl-tRNA, anticodon UCA, that specifically recognizes the UGA codon is required for selenocysteine incorporation into formate dehydrogenase. A eukaryotic UGA suppressor tRNA with UCA anticodon that accepts serine and is phosphorylated to O-phosphoseryl-tRNA may have a corresponding function in glutathione peroxidase synthesis. Other factors required for the unusual usage of the in-frame UGA codons to specify selenocysteine incorporation and the biochemical mechanism involved in distinguishing these from normal UGA termination codons are discussed. PMID- 2978459 TI - T cell development in B cell deficient mice. III. Restriction specificity of suppressor T cell factor(s) produced in mice treated chronically with rabbit anti mouse mu chain antibody. AB - A role for Igh linked genes and the idiotypes they encode has been implicated in the activity of a variety of T cell subpopulations. Idiotype restricted T cell function has been observed for helper and suppressor cell populations. The finding that T cell receptor genes are distinct from B cell receptor (Igh) genes strongly argues against a direct role for immunoglobulin genes in the determination of the T cell repertoire. Nevertheless, idiotypic Ig determinants may play an indirect role in influencing the ultimate composition of the T cell repertoire. One approach to this question involves evaluation of T cell activity upon development in an immunoglobulin deficient environment. The availability of antigens which elicit T cell and antibody responses characterized by the expression of dominant crossreactive idiotypes under the control of Igh genes provides an ideal approach to investigate the basis for the expression of Igh like structures on T cells and the concomitant functional genetic restrictions they determine. Thus, we have prepared B cell deficient mice by continuous treatment, beginning at birth, with rabbit anti-mouse IgM. The network which comprises the suppressor T cell response to azobenzenearsonate (ABA) was then examined in normal and anti-mu treated mice to assess what role, if any, immunoglobulin encoded determinants play in influencing the composition of the peripheral T cell pool. The results clearly demonstrate that the absence of Ig+ B cells leads to major alterations in the composition of the T cell repertoire. Anti-mu treated, but not normal rabbit Ig treated, mice produce TsF1 which fails to suppress cytotoxic T lymphocyte or helper T cell responses of normal syngeneic mice, yet efficiently suppresses those of syngeneic anti-mu treated recipients. Reciprocally, normal TsF1, though suppressive in normal Igh-1 syngeneic recipients, fails to affect the development of responses in anti-mu treated syngeneic mice. TsF1 obtained from anti-mu treated mice is antigen-specific. Testing of anti-mu TsF in a variety of normal or anti-mu treated recipients reveals no MHC restrictions. In marked contrast, anti-mu TsF reflects a novel pattern of Igh functional restrictions. The observed Igh restrictions were found to map to the idiotype encoding VH regions of the Ig heavy chain gene cluster (Igh-VH). The results demonstrate that T cell maturation in the virtual absence of environmental immunoglobulin can lead to profound changes in the composition of the T cell compartment. The means by which the absence of Ig encoded determinants leads to such changes is speculated upon. PMID- 2978460 TI - The putative I-Jk- strain AKR/J synthesizes I-Jk+ molecules: implications for Jt gene control of I-J expression. AB - At present, the genetic basis for control of murine I-J determinants is unknown. On one hand, it is clear that polymorphism in I-J molecules is controlled by genes mapping in the I region of the H-2 gene complex on chromosome 17. On the other hand, molecular genetic studies provide evidence that I-J molecules are not encoded by I region genes. Although formal proof of the latter must await isolation and characterization of I-J structural genes, these observations are compatible with the concept that I-J molecules are encoded by non-H-2 genes, but the expression of these non-H-2 genes is regulated or influenced by I region genes. Recent studies by Hayes et al. provide evidence for non-H-2 control of the cell surface expression of I-Jk determinants in strain AKR/J. This strain typed I Jk-, as judged by complement dependent cytolysis with monoclonal I-Jk antibodies. Studies with recombinant inbred and congenic strains suggested that the I-Jk- phenotype in strain AKR/J was controlled by a gene (Jt) mapping on chromosome 4. Based on these observations and studies with F1 hybrids and numerous other strains, Hayes et al. concluded that interaction between the Jt gene and an H-2 gene on chromosome 17 (probably E beta or E alpha) regulates the production and expression of I-Jk molecules, and hypothesized that I-Jk epitopes may reside on Jt modified Class II molecules or on the Jt gene product.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978462 TI - An investigation into non-fluency in Down's syndrome. PMID- 2978461 TI - Nebulization versus injection in ambulatory treatment of acute asthma: a comparative study. AB - Nebulization treatment of acute asthma with terbutaline and ipratropium bromide, given either separately with a 30-minute interval or combined as single inhalation, was compared with injection treatment with a combination of terbutaline given subcutaneously and theophylline given intravenously. Seventy seven episodes of acute asthma were studied. Nebulization treatment gave the same degree of bronchodilation as the injections, both immediately after treatment [measured as increase in forced expiratory volume in 1 second (FEV1) in peak expiratory flow rate (PEFR) and improvement of dyspnoea] and during the following 6 days (measured by PEFR recordings at home). The injection treatment caused a moderate increase in heart rate, whereas no circulatory side-effects were noted during nebulization treatment. Administration of ipratropium bromide 30 minutes after terbutaline was not more effective than the combination of both substances as a single nebulization. PMID- 2978463 TI - [Effect of burn infection on the functions and structure of rabbit erythrocyte membrane]. PMID- 2978464 TI - Collagen network remodelling and diastolic stiffness of the rat left ventricle with pressure overload hypertrophy. AB - This study had two objectives: (a) to determine the accumulation of collagen and its structural remodelling in the hypertrophied rat left ventricle after 4 and 8 weeks of abdominal aorta banding; and (b) to correlate these findings with the diastolic stress-strain relation of the intact myocardium. In comparison to age and sex matched controls, the collagen volume fraction of the hypertrophied myocardium after 4 and 8 weeks of aortic banding increased significantly from 3.5(SD1.0)% to 7.8(4.2)% and 6.2(2.0)% respectively. This accumulation of collagen, or fibrosis, occurred in the absence of myocyte necrosis. Scanning electron microscopy showed increased density and thickness of the collagen weave and tendons. At 4 weeks, light microscopy showed interstitial oedema and disrupted collagen fibrils. Left ventricular diastolic stress-strain relations of both pressure overload groups were significantly steeper than that of the control group. Thus the response of the interstitium to the hypertrophic process that accompanies abdominal aorta banding is a complex process that includes a structural remodelling of the fibrillar collagen matrix and the early appearance of interstitial oedema, each of which may contribute to a rise in the passive stiffness of the intact myocardium. PMID- 2978465 TI - A simple technique for producing supravalvular aortic stenosis in animals. AB - A safe and reproducible technique to create supravalvular aortic stenosis was developed, which avoids many of the difficulties encountered in the production of aortic stenosis. Dogs were anaesthetised and artificially ventilated. The chest was opened and the venae cavae were encircled with umbilical tapes. The ascending aorta was then encircled by a 1.5-2 cm wide, 6-7 cm long dacron patch, venous return was stopped by tightening the tapes, and a J-shaped clamp applied to the ascending aorta at the dacron patch. Two layers of continuous mattress suture were placed adjacent to the clamp, plicating the aortic diameter by about 50%. After releasing the clamp and restoring normal venous return, left ventricular (LV) and aortic (AO) pressures were measured. Subsequently, one or two deep mattress sutures were placed below the running mattress sutures to increase the stenosis and to obtain the desired gradient. The LV-AO systolic pressure gradients obtained immediately after the operation ranged from 40 to 75 mm Hg. Two to 6 months after the operation the pressure gradients ranged from 50 to 200 mm Hg. Left ventricular to body weight ratios were 6.41 (SEM 0.26) v 4.24(0.20) for the controls. Heart weight to body weight ratios were 8.37(0.35) v 5.65(0.33). LV end diastolic pressures were normal. This technique can be used either in puppies or adult animals. The problem of aortic rupture is eliminated. The pressure gradient can be easily controlled during the operation and reproducible LV hypertrophy can be obtained in a shorter time than with aortic banding of puppies. PMID- 2978466 TI - Bioactive LH in women with polycystic ovaries and the effect of gonadotrophin suppression. AB - Discrepancies between levels of bioactive LH (B-LH) and immunoreactive LH (I-LH) in polycystic ovarian syndrome (PCO) have been reported previously. Serum levels of I-LH, B-LH (by dispersed Leydig cell assay), FSH, oestradiol (E2) and progesterone (Prog) were measured once to three times weekly over 4 weeks in 13 women with classical clinical, ultrasound and endocrine features of PCO. Eleven women attending for infertility but whose profiles when studied three times weekly by combined endocrine and ultrasound assessment were normal and ovulatory served as controls. Seven of the women with PCO were evaluated during and after 3 weeks suppression with ethinyloestradiol (30 micrograms) plus 150 micrograms either of desogestrel or levonorgestrel; two were given both treatments. Both I LH and B-LH levels were higher in PCO patients (20 +/- SD 5 U/l and 46 +/- 9 U/l respectively, P less than 0.0001), compared with all phases of the normal cycles except the mid-cycle peak. The B-LH to I-LH (B:I LH) ratio in PCO patients (2.5 +/- 0.7) was higher than in all the control cycle phases (P less than 0.05). I LH, B-LH, B:I LH ratio, FSH and E2 were all suppressed from the second week of oestrogen-progestogen treatment (P less than 0.01) and returned gradually to pretreatment levels by the third or fourth week after suppression. The LH and FSH levels and B:I LH ratio in PCO patients during suppression were comparable with levels in the early and mid-follicular phases of control cycles but the LH/FSH ratio remained significantly raised (P less than 0.01) at 2.3 +/- 0.7.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978467 TI - Pityriasis rubra pilaris and vitiligo in Down's syndrome. PMID- 2978468 TI - Keratoderma of myxoedema. PMID- 2978470 TI - 1988 fountain of youth. PMID- 2978469 TI - Haemorrhagic pompholyx in bullous pemphigoid. PMID- 2978471 TI - Donated dental services program takes off in New Jersey. PMID- 2978472 TI - Assessing dental manageability of handicapped children. PMID- 2978473 TI - [A comparative study of apical hypertrophy with giant negative T waves and non obstructive hypertrophic cardiomyopathy: evaluation by computed tomography, echocardiography and left ventriculography]. AB - To assess the relationships between patterns of left ventricular hypertrophy and giant negative T waves (greater than 15 mm, GNT), 24 patients with GNT were categorized in two groups: Group A consisted of 12 patients with GNP but without left ventricular hypertrophy (greater than 13 mm at the chorda level of the interventricular septum and/or left ventricular posterior wall by UCG, LVH); and group-B consisted of 12 patients with GNT and LVH. Twelve patients with LVH but no GNT compromised as group-C. Left ventriculography (LVG) was performed, and left ventricular mass (LVM) and apical hypertrophy were assessed by CT. The configurations of the end-diastolic left ventricle by LVG (RAO 30 degrees) were as follows, by group: A: spade in eight patients and papillary muscle hypertrophy (PMH) shape in four. B: spade in five, PMH in four, banana in one and oval in two. C: banana in nine and oval in three. The LVM (g/BSAm2) were as follows; A: 121.5 +/- 14.9, B: 130.5 +/- 14.8, C: 190.6 +/- 42.1 (C greater than A, B, p less than 0.001). The apex/base ratios calculated by CT were as follows; A: 1.43 +/- 0.31, B: 1.21 +/- 0.19, C: 1.09 +/- 0.20 (C greater than A, p less than 0.05). The left ventricular mass in patients with GNT was less than that of those without GNT. Giant negative T waves were most often associated with apical hypertrophy. PMID- 2978474 TI - [Natural course of hypertrophic cardiomyopathy: clinical, hemodynamic and echocardiographic features in the end stage]. AB - We investigated the natural course of 59 patients with hypertrophic cardiomyopathy (HCM) in follow-up periods of one to 13 years and analyzed the clinical, hemodynamic and echocardiographic parameters to determine the factors influencing the prognosis. Among these patients, 44 (75%) remained stable in a compensated condition with or without medications. Five patients died suddenly and two died of congestive heart failure. Heart failure developed in another eight. At the initial evaluation, these 15 patients had high left ventricular end diastolic pressure (mean: 22 +/- 8 mmHg) significantly higher than that of 44 compensated patients (mean: 13 +/- 6 mmHg, p less than 0.001). There were no differences in age at the initial evaluation between compensated and end stage groups. Atrial fibrillation occurred persistently in three and transiently in two among ten patients with heart failure during follow-up periods. Ventricular dimensions and systolic function did not statistically differ between the two groups. However, six patients with heart failure had cavity dilatation and deteriolated ventricular contractile function at the initial evaluation. Four of them did not show any change in left ventricular hypertrophy, but the regional thinning of the wall was observed in the remaining two. There were no characteristic features to predict sudden death in the clinical, hemodynamic or echocardiographic parameters. Thus, increased left ventricular end-diastolic pressure, atrial fibrillation, left ventricular dilatation and the regional thinning of the left ventricular wall are useful predictive markers for poor prognosis in HCM. PMID- 2978475 TI - [Clinical pictures and prognosis of mitral valve prolapse in the middle-aged and old-aged patients]. AB - The age as an important factor influencing the prognosis of mitral valve prolapse (MVP) was assessed by comparing two age groups; 26 cases whose ages were younger than 49 years, and 27 cases more than 50 years. 1. The aged group had higher incidence of dyspnea and enlarged heart most probably due to insidious heart failure. 2. The aged group frequently had an apical pansystolic murmur and enlarged left atrium, indicating the progress of mitral regurgitation. These observations suggest that the careful follow-up observation is mandatory for patients with MVP of advanced age. PMID- 2978476 TI - Digital calibrator for intravital velocimetry. PMID- 2978477 TI - [Repair of a gap in the abdominal wall using intraperitoneal Seblon mesh]. PMID- 2978478 TI - [The dam in pedodontics: placement technic]. PMID- 2978479 TI - [The dental assistant in pedodontics]. PMID- 2978480 TI - [Oral hygiene and the handicapped. Trial of a controlled program in a group of disabled rural residents]. PMID- 2978481 TI - [Structure and function of proteoglycans]. PMID- 2978482 TI - Can something be done for patients with liver metastases for colorectal cancer? PMID- 2978483 TI - The management of acne vulgaris. PMID- 2978484 TI - Coronary thrombolysis. PMID- 2978485 TI - [Coronary angioplasty]. PMID- 2978487 TI - [Oral surgery during anticoagulant therapy]. PMID- 2978486 TI - [Prevention and therapy of oral manifestations in leukemic patients]. PMID- 2978489 TI - [Stomatological aspects of AIDS]. PMID- 2978488 TI - [Oral surgery in patients with coagulation disorders and hemophilia]. PMID- 2978490 TI - [Dental interventions in haemorrhagic diatheses and long-term therapy with anticoagulants]. PMID- 2978491 TI - [First experiences with a stomatological dispensary for cerebrally disabled patients]. PMID- 2978492 TI - Effect of exhaustive exercise on liver mitochondrial function in the rat. AB - The oxidative and phosphorylative function of rat liver mitochondria after exhaustive exercise was investigated. The stimulation of state 4 respiration (without ADP) with NADH and FADH2 dependent substrates was demonstrated. The reduction in RCR ratio (the rate of oxidation in state 3/the rate of oxidation in state 4) and enhanced activity of oligomycin sensitive ATP-ase was also found. The results suggest an inhibition of liver mitochondrial phosphorylative activity in rats exercised till exhaustion. PMID- 2978493 TI - Abdominal wall defects and obstructive uropathies. Aetiology and management. AB - Gastroschisis and omphalocele (exomphalos) are embryologically distinct, and for this reason each has its own distinct constellation of associated anomalies. The perinatal management of both gastroschisis and omphalocele addresses the controversy as to the best mode of delivery that has been expressed in the literature with regard to gastroschisis and has exercised many of those involved in antenatal diagnosis in the case of omphalocele. The post-delivery management of both of these defects is briefly discussed. Controversies surrounding the aetiology of renal dysplasia and whether urinary tract obstruction is a cause of dysplasia or an anomaly associated with dysplasia are also discussed. The options for therapy and the indications for an in utero shunting procedure are outlined as well as the controversy centred around the utility of an in utero shunt. The need for a prospective randomised trial is emphasised, together with the difficulty that is raised by the need to choose a limited number of prospective indices on which treatment decisions in such a trial must be based. The need for a long-term follow-up of all patients entered in such a trial is emphasised. PMID- 2978494 TI - [Local anesthetics: how to treat patients with suspected allergy]. PMID- 2978495 TI - Enoximone in intensive care. Proceedings of a symposium. Brussels, Belgium, 21st March 1988. PMID- 2978496 TI - Enoximone in heart failure: mechanisms of action. PMID- 2978497 TI - Comparative effects of enoximone and nitroglycerin on left ventricular performance and regional wall motion in ischaemic cardiomyopathy. PMID- 2978498 TI - Effects of enoximone in patients with cardiac failure after myocardial infarction. PMID- 2978499 TI - Comparison of haemodynamic efficacy of enoximone and dobutamine in coronary surgery patients. PMID- 2978500 TI - Effects of enoximone in patients with low ejection fraction undergoing coronary artery bypass graft surgery. PMID- 2978501 TI - Efficacy of enoximone in the management of refractory low output states following cardiac surgery. PMID- 2978502 TI - A study of the comparative effects of enoximone and dobutamine on urinary output after open heart surgery. PMID- 2978503 TI - Use of enoximone in patients awaiting cardiac transplant. PMID- 2978504 TI - Enoximone, a pharmacological bridge to transplantation. PMID- 2978505 TI - [Repair of a large defect of the abdominal wall by myocutaneous flap of m. gracilis with blood vessel anastomosis--a case report]. PMID- 2978506 TI - [The effect of two resuscitation regimens on cardionatrin during burn shock]. PMID- 2978507 TI - [Changes in immunological function of red cells in burn patients and their significances]. PMID- 2978508 TI - Verdicts differ in AIDS cases with patients. PMID- 2978509 TI - New York City dentist wins AIDS discrimination dispute. PMID- 2978510 TI - [Tooth extraction in coagulopathic patients]. PMID- 2978511 TI - Marfan syndrome: dental problems and management. PMID- 2978512 TI - [Correlation between anxiety and pain in ambulatory dental patients]. PMID- 2978513 TI - [General anesthesia in dentistry for patients at hepatic, renal and metabolic risk]. PMID- 2978514 TI - The Groningen Social Disabilities Schedule: development, relationship with I.C.I.D.H., and psychometric properties. AB - Most of the existing instruments for the assessment of social dysfunctioning of psychiatric patients are merely lists of precoded items, leaving little or no room for posing supplementary questions. In this article a new, semi-structured questionnaire--the Groningen Social Disabilities Schedule (GSDS)--is described, in which this shortcoming is met. The development of the GSDS is based on social role theory and on the existence of a hierarchy in social disabilities, demonstrated in an earlier World Health Organization study. Compatibility was sought with the International Classification of Impairments, Disabilities and Handicaps (ICIDH), a trial-classification of the World Health Organization, to be used in research on the consequences of illness. It is concluded that the inter rater reliability of the GSDS is good. To a large extent the hierarchy proved to hold true for this instrument. Some implications for future research and for the ICIDH are discussed. PMID- 2978515 TI - The biological evolution of the Leeds-Keio ligament in the human knee. A histological and ultrastructural study. AB - The authors report their experience in the reconstruction of the anterior cruciate ligament using the Leeds-Keio ligament in 12 patients, 7 by open arthrotomy and 5 by an arthroscopic technique. The biological evolution of the new ligament was studied macroscopically and histologically in the first 6 cases treated by open arthrotomy. Arthroscopic monitoring and biopsy was carried out 4 to 10 months after operation. In 5 cases the new ligament appeared to be intact and histologically vital, with proliferation of new tissue along the new ligament. There were broken filaments of dacron hanging from the intercondylar cavity in only one case. The first macroscopic, histological and ultrastructural results show that despite a few limitations the Leeds-Keio ligament has the features required for replacement of the anterior cruciate ligament, although it will be necessary to wait 4 or 5 years before fully evaluating the durability and function of the ligament and the best operative method. PMID- 2978516 TI - The colony-stimulating factor 1 (CSF-1) receptor (c-fms proto-oncogene product) and its ligand. AB - Alterations in genes that function in normal growth and development have been linked to malignant cell transformation. The mononuclear phagocyte colony stimulating factor (CSF-1 or M-CSF) is a polypeptide growth factor synthesized by mesenchymal cells, which stimulates the survival, proliferation, and differentiation of haematopoietic cells of the monocyte-macrophage series. Multiple forms of soluble CSF-1 are produced by proteolytic cleavage of membrane bound precursors, some of which are stably expressed at the cell surface. The c fms proto-oncogene encodes the CSF-1 receptor, which is composed of an extracellular ligand-binding domain linked by a single membrane-spanning segment to a cytoplasmic tyrosine-specific protein kinase domain. Whereas the tyrosine kinase activity of the normal receptor is stimulated by CSF-1, mutations in the c fms gene can constitutively activate the kinase to provide growth-stimulatory signals in the absence of the ligand. Oncogenic activation of the c-fms gene product appears to involve removal of a negative regulatory tyrosine residue near the carboxyl terminus of the receptor and one or more additional mutations that may simulate a conformational change induced by CSF-1 binding. Expression of the human c-fms gene in mouse NIH-3T3 cells confers a CSF-1 stimulated growth phenotype, indicating that receptor transduction is sufficient for fibroblasts to respond to a haematopoietic growth factor. In contrast, the v-fms oncogene induces factor-independent growth and tumorigenicity in factor-dependent myeloid cell lines, and contributes to the development of proliferative disorders of multiple haematopoietic lineages when introduced into murine bone marrow progenitors. Aberrant expression of an endogenous c-fms gene secondary to proviral insertion and transcriptional activation has also been implicated in virus-induced myeloblastic leukaemia in mice. The c-fms and CSF-1 genes have been mapped on the long arm of human chromosome 5, a region that frequently undergoes interstitial deletions in certain haematopoietic disorders including acute myelogenous leukaemia. The study of CSF-1 and its receptor should provide information concerning the role of tyrosine kinases in regulating the normal growth and differentiation of haematopoietic cells and in contributing to their malignant transformation. PMID- 2978517 TI - Structure and function of Fc receptors on macrophages and lymphocytes. AB - Cell surface receptors for the Fc portion of immunoglobulin confer on most cells of the immune system the ability to communicate with the humoral antibody response. These Fc receptors are known to be particularly important for the function of various effector cells, such as macrophages, since they are involved in mediating a variety of activities including endocytosis, antibody-dependent cellular cytotoxicity, and triggering the release of potent inflammatory agents. Over the past few years, a considerable amount has been learned about the structure and functions of the Fc receptors expressed by murine and human cells, due to the availability of specific anti-receptor antibodies and the isolation of Fc receptor cDNA clones. In general, these receptors are transmembrane proteins whose extracellular domains contain two immunoglobulin-like regions and are thus members of the immunoglobulin gene family. Their domain structure consists of a glycosylated extracellular domain, a single membrane-spanning segment, and a relatively long cytoplasmic domain. The cytoplasmic tails exhibit a surprising degree of variation in length and amino acid sequence. This review summarizes some recent information concerning the structure and expression of the Fc receptors found on murine and human macrophages and lymphocytes. Particular attention is paid to the functional activities of these receptors, and the possible relationship between receptor function and receptor structure. PMID- 2978518 TI - C3 receptors on macrophages. AB - The complement receptors on macrophage are responsible for their binding and ingestion of opsonized targets. The two established receptors are CR1, which recognizes C3b, and CR3, which recognizes iC3b, the natural product of C3b from cleavage by the complement control protein factor I and its cofactors. CR1 belongs to a group of proteins that contain a structural element characterized by its size of 60-65 amino acids, and four conservatively positioned cysteines, which engage in a self-contained 1-3, 2-4 disulphide arrangement. This structural unit is called SCR (short consensus repeat) and is found in the complement proteins C1r, C1s, C2, factor B, factor H, C4BP, DAF, MCP and CR2, each of which interacts with some cleavage products of C3 and/or C4. CR1 has 30 SCR units accounting for its entire extracellular structure. It has a transmembrane segment and a small cytoplasmic domain. CR3 is a heterodimer containing an alpha and beta subunit held together by non-covalent forces. The beta subunit is also found in the two leukocyte antigens, LFA-1 and p150,95, which have alpha subunits distinct from that of CR3. The beta subunit contains 56 cysteine residues, 42 of which lie in a span of 256 residues immediately adjacent to the transmembrane segment. It shares extensive sequence homology with subunits of membrane protein complexes that bind fibronectin and vitronectin, implicating that they all belong to an extended set of surface adhesion molecules not restricted to the immune system. p150,95 is also expressed on macrophages and it has iC3b binding activity. It also shares some functional properties with CR3 as an adhesion surface molecule. PMID- 2978519 TI - Adhesion-promoting receptors on phagocytes. AB - Phagocytes express a family of structurally related receptors, LFA-1, CR3, and p150,95, that mediate adhesion of leukocytes to a variety of cells and surfaces. LFA-1 mediates the binding of killer T cells to targets, CR3 mediates binding of phagocytes to iC3b-coated surfaces and to endothelial cells, and LFA-1, CR3, and p150,95 each mediate the binding of bacterial lipopolysaccharide. Here we review the structure and function of each of these receptors and present evidence that they are related to a larger class of adhesion-promoting receptors called integrins. Of particular emphasis are observations that the capacity of these receptors to promote adhesion is strongly and reversibly modulated by both soluble and surface-bound stimuli. We review this form of regulation and present evidence that changes in the binding activity of adhesion-promoting receptors is accomplished by changes in the two-dimensional distribution of receptors in the plane of the membrane. Inactive receptors are randomly distributed in the membrane, and their ability to bind a ligand-coated surface is enabled by a ligand-independent movement into small clusters. The implications of these structural features are discussed. PMID- 2978520 TI - Ethical ramifications of refusing to treat patients with AIDS or patients who are HIV seropositive. PMID- 2978521 TI - Fatal pulmonary embolus in a patient treated with Marvelon. PMID- 2978522 TI - [Yeast flora of the intestinal tract in long-lived residents of Abkhaziia]. PMID- 2978523 TI - [Antagonism phenomenon in killer yeast Saccharomyces cerevisiae, isolated from the intestinal tract of long-lived residents of Abkhaziia]. PMID- 2978524 TI - [Dentists and AIDS]. PMID- 2978525 TI - [Identification of degenerative processes as a cause of lumbar pain syndrome]. PMID- 2978526 TI - [The incidence of low back pain in hotel and restaurant workers]. PMID- 2978527 TI - [Low back pain in pregnancy]. PMID- 2978528 TI - [The effect of the cervical and lumbar syndrome on work capacity]. PMID- 2978529 TI - [Acute lumbar syndrome in tourists]. PMID- 2978530 TI - [Roundtable on prophylaxis of complications and consequences for the dentist and radiotherapist in patients undergoing radiotherapy for upper digestive or airway neoplasms or Hodgkin's disease]. PMID- 2978531 TI - [Antibiotic prophylaxis in dentistry]. PMID- 2978532 TI - [Blood pressure measurement: involvement of South African dentists]. PMID- 2978533 TI - [Surgical intervention in heart transplant patients]. PMID- 2978534 TI - [Sensometric function therapy by pediatrician and dentist in orofacial malfunction]. PMID- 2978535 TI - Interaction of membrane-bound Fc gamma receptors with the cytoskeletal matrix. AB - Binding of ligands to cell surface receptors may induce an interaction of the receptors with the cell cytoskeleton. This interaction may decrease the solubility of the receptors in nonionic detergents. We studied effect of binding of various 125I-labeled immunoglobulin ligands to Fc gamma receptors on guinea pig peritoneal macrophages and human placental syncytiotrophoblast plasma membranes on an interaction of these receptors with the cytoskeletal matrix. The receptor-cytoskeleton association was evaluated by measurement radioactivity of bound ligands in pellets and supernatants obtained after lysis of cells or membranes in a nonionic detergent NP-40. Binding of soluble immune complexes or crosslinking of IgG bound induces much stronger insolubilization of the receptors than binding of monomeric or aggregated IgG. It shows that the interaction of the receptors with the cytoskeletal matrix strongly depends on the degree of cross linking of the Fc gamma receptors by ligands bound. The observed effects were IgG Fc region-specific. Isolated, purified putative Fc gamma receptors from guinea pig peritoneal macrophages and from human placental syncytiotrophoblast plasma membranes do not interact with free G or F actin. We also studied association of the guinea pig peritoneal macrophage Fc gamma receptor with the cytoskeleton, before and after shedding of macrophage membrane proteins. The results obtained showed that the macrophages have only one class of Fc gamma receptors interacting with the cytoskeletal matrix. Effect of a cytoskeleton-destabilizing buffer and DNAse I on release of the receptors from the cytoskeleton suggests that insolubilization of ligand-Fc gamma R complexes was caused, at least partially, by an interaction of the receptors with actin filaments in the cytoskeleton. The results presented in this paper suggest that the cytoskeleton might play a role in transmission of signals from Fc gamma receptors to the cells. They underline the role of immune complexes as physiological ligands for Fc receptors and correlate well with activation of cells via their Fc receptors (e.g. superoxide burst) observed by other authors after treatment of the cells with immune complexes, but not with monomeric or aggregated IgG. PMID- 2978536 TI - Relationships between structure and function in the Fc receptor family. PMID- 2978537 TI - Modulation of prostaglandin and leukotriene biosynthesis. PMID- 2978538 TI - T cell receptors: structure and genetics. PMID- 2978539 TI - Lymphocyte subsets and activation. PMID- 2978540 TI - The therapeutic use of monoclonal antibodies. PMID- 2978541 TI - A clinical study of topical mucopolysaccharides & polydeoxyribonucleoprotein (Foltene) therapy in alopecia. AB - We performed clinical trials to evaluate the therapeutic effects of Foltene in patients of the several types of hair fallings. Thirty patients with male pattern baldness, alopecia areata and seborrheic alopecia were included in this study. Foltene was applied every other day for 40 days, and followed by maintenance therapy of twice application a week. The duration of whole therapy was 6 months. We conclude that Foltene is an effective and agent for male pattern baldness, alopecia areata and seborrheic alopecia from the following results. Ten patients with male pattern baldness was treated with Foltene for 6 months. Foltene had therapeutic effects of 50% in hair regrowth, 70% in decreased hair falls, 30% in decreased dandruff, 50% in decreased seborrhea. Thirteen patients with alopecia areata was treated with Foltene for 6 months. Foltene had therapeutic effects of 61.6% in hair regrowth, 53.9% in decreased in hair falls, 53.9% in decreased dandruff, 77.0% in decreased seborrhea. Seven patients with seborrheic alopecia was treatment with Foltene for 6 months. Foltene had therapeutic effects of 85.8% in hair regrowth, 57.2% in decreased hair falls, 42.9% in decreased dandruff, 85.8% in decreased seborrhea. The degree of therapeutic success was related to the duration of therapy. The side effects were as followed: itching sensation developed in 2 patients (6.7%); tingling sensation in 3 patients (10.0%); burning sensation in 1 patient (3.3%); erythema in 3 patients (10.0%). PMID- 2978542 TI - Toward a social ecology of the home-care household. AB - I examined the relation between household parameters of size, composition, and temporal variability and indicators of care-giver and elder well-being. Two discrete groups of 20 care-giver-elder pairs were included in the study: (a) a group in which elders were moderately to severely cognitively impaired, but physically well, and (b) a group in which elders were moderately to severely physically disabled, but cognitively unimpaired. Care givers and elders were interviewed and assessed in their homes, and the care giver completed a 3-day log of home-care activities. Results indicated that care-giver and elder well-being are related to opposite values of the same household parameters in the two groups. The study demonstrates the utility of a social-ecological framework for research on home care, and suggests that the degree of fit of any particular household arrangement is relative to the competence-disability profile of the elder in care. PMID- 2978543 TI - Stressful situations in caregiving: relations between caregiver coping and well being. AB - We examined ways in which caregivers cope with stressful caregiving situations and the relations between coping strategies and caregivers' psychosocial well being. Respondents were 58 family caregivers to patients discharged from a rehabilitation hospital. Caregivers identified a recent stressful event in caregiving and indicated strategies used to cope with this event. After controlling for patients' impairment level, analyses indicated that caregivers engaging in more escape-avoidance coping reported greater depression and more conflict in their personal relationships. Those using more positive reappraisal demonstrated greater positive affect. Younger caregivers, many of whom were women, used more avoidance strategies. Results have implications for therapeutic interventions with family caregivers. PMID- 2978544 TI - Preliminary characterization of two glucan synthetase preparations and their reaction products from Candida albicans. AB - Two glucan synthetase preparations from Candida albicans were obtained by lysis of regenerating protoplasts (enzyme A) or mechanical breakage of yeast cells (enzyme B). Enzyme A was insensitive to EDTA or GTP but it was stimulated by a combination of both agents. Enzyme B was inhibited by EDTA, this inhibition being released by increasing the concentration of the chelating agent or by addition of GTP to the assay mixtures. Enzyme A was further activated by glycerol and sodium fluoride. The reaction products were characterized as linear beta-1,3-linked glucans on the basis of their resistance to periodate and susceptibility to beta glucanases. In both cases the "in vitro" synthesized radioactive chains were added to the non-reducing end of cold, performed glucan or to and acceptor other than glucan. At least, part of the preformerd glucan chains of enzyme A, but no those of enzyme B, showed a free reducing terminal. On the basis of the origin of both enzyme preparations it is suggested that glucan molecules are synthesized while bound to an acceptor of a different nature which is subsequently excised. PMID- 2978545 TI - TCR beta chain expression during human thymic development. PMID- 2978546 TI - [The plastic surgery for adiposis of the abdominal wall--report of 9 cases]. PMID- 2978547 TI - [Orodental intervention before and after radiotherapy of the oral cavity]. PMID- 2978548 TI - [Robert Bunon (1702-1748). A precursor to pedodontics]. PMID- 2978549 TI - [Dacron mesh and surgical therapy of inguinal hernia]. PMID- 2978550 TI - [Dental care for children and youths in Zurich]. PMID- 2978551 TI - [The history of pedodontics in Budapest]. PMID- 2978552 TI - [Results of systematic pedodontic and preventive dental care in Visegrad]. PMID- 2978553 TI - [The work of our orthodontic department]. PMID- 2978554 TI - [The Castillo-Morales palatal plate]. PMID- 2978555 TI - [Dentistry for handicapped patients]. PMID- 2978556 TI - [Dental prevention for the handicapped]. PMID- 2978557 TI - [The psychology of parents of uncooperative handicapped children]. PMID- 2978558 TI - [Dentoperiodontal conditions in a group of handicapped residents in rural areas]. PMID- 2978559 TI - [Dentistry for handicapped patients]. PMID- 2978560 TI - [Prosthetic rehabilitation of uncooperative patients]. PMID- 2978561 TI - [Advantages of conscious sedation in the handicapped]. PMID- 2978562 TI - [Dental treatment with general anesthesia for neuropsychiatric patients]. PMID- 2978563 TI - [Dental care under anesthesia for the handicapped]. PMID- 2978564 TI - [Dental treatment for handicapped patients: medico-legal considerations]. PMID- 2978565 TI - [Didactic amplification]. PMID- 2978566 TI - Approach to patients with degenerative disorders of the nervous system. PMID- 2978567 TI - Survey of dental need among veterans with severe cognitive impairment. PMID- 2978568 TI - Dental treatment planning for patients with severe cognitive impairment. PMID- 2978570 TI - Argument against providing dental care for the severely cognitively impaired patient. PMID- 2978569 TI - Establishing a preventive dentistry program in a long-term health care institution. PMID- 2978571 TI - Argument in favor of providing dental care for the severely cognitively impaired patient. PMID- 2978572 TI - Dental care for the cognitively impaired: an ethical dilemma. PMID- 2978573 TI - Dental management of a patient with cognitive impairment: a case report. PMID- 2978574 TI - The evaluation and management of a dental patient with a bleeding disorder. PMID- 2978575 TI - Evaluation and management of the dental patient with cancer II: Patients receiving head an neck radiation therapy. PMID- 2978576 TI - [Osteoradionecrosis: risks and prevention in dentistry]. PMID- 2978577 TI - [Clinical characteristics of bleeding disorders]. PMID- 2978578 TI - A history of the British Paedodontic Society. Part V. Proceedings and other publications. PMID- 2978579 TI - Ethical considerations of appropriate versus comprehensive dental care for patients in nursing homes. PMID- 2978580 TI - Predictors of dental behavior of disabled children. PMID- 2978581 TI - Attitudes toward dental care for children with cancer: results of a maternal questionnaire. PMID- 2978583 TI - Can biomedical engineering support preventive medicine? PMID- 2978582 TI - [The induction of suppressors of the humoral immune response in experimental staphylococcal infection]. PMID- 2978584 TI - Development of a microprocessor-based adaptive technique for blood pressure measurements. AB - The paper introduces a new microprocessor-based adaptive technique for the indirect measurement of the systolic and diastolic pressure in humans. The technique is based upon a statistically consistent relationship between the amplitude of the pulsative pressure waveform at the systolic and diastolic points and the amplitude of pulse signals detected when the artery is fully occluded. An adaptive measurement philosophy has been implemented in the design of an electronic analog-digital sphygmomanometer which, in addition to a pressure transducer, contains suitable electronic instrumentation for processing and displaying the electronic signals. A dedicated microprocessor is used to store statistical relations and control the operation of the device. Verification of overall system accuracy is accomplished via direct comparison with manual auscultatory measurements. Clinical testing of a prototype indicates satisfactory performance; measurement errors are maintained well within proposed standards for automated sphygmomanometers. PMID- 2978585 TI - Magnetic field of atrial depolarization. AB - The isomagnetic maps of normal subjects and patients with right and left atrial overloading were recorded to determine the characteristic features of the magnetic field of atrial depolarization. The isomagnetic maps examined in this study indicated the instantaneous current source, which specifically localizes the current sources due to the right and left atria, respectively. The magnetic field recorded with a second derivative gradiometer clearly detected the cardiac current source from the right atrium, which is located close to the anterior chest wall, thus this method improved the diagnostic sensitivity for right atrial overloading. In patients with left atrial overloading, the isomagnetic map showed multiple dipoles due to the right and left atria, respectively, which are difficult to be detected by the electrocardiogram or isopotential map. These results suggest that the magnetocardiogram provides useful information on the current source to supplement information obtained by the conventional electrocardiogram. PMID- 2978586 TI - Automated data collection and presentation in the operating room. AB - An 'Operating Room Data Integration System', is described which is used to collect, present and archive all important physiological parameters during open heart surgery. The system requires very little attention, and provides an easy to understand and coherent interface to the user. The system is adaptable to a large extend and thus data can be presented to the user in a manner, with which he or she is already familiar. Simple drivers can be written to enable connection of the system to almost any other piece of medical equipment, if the latter provides an analog or digital, output signal. Automatic logging of the acquired signals is then possible. PMID- 2978587 TI - EEG mapping: current status and future prospects. AB - EEG mapping is the topographical display of parameters evaluated from multichannel EEG recordings. Different problems connected with EEG mapping are: number of electrodes, interpolation between electrodes, types of reference and statistical treatment of maps. These problems are discussed briefly and examples are given. To complete this report, a short historical background along with some comments about the clinical settings have been provided and attention given to future prospects. PMID- 2978588 TI - Technology in health care: the social impact and economic cost. AB - The contribution of technology to longevity and the quality of life has been substantial during the twentieth century. In the past two decades, technology employed in the care of hospitalized patients has been responsible for most of the cost by which the medical inflation rate (Medical Economic Index) exceeds the consumer price index. In most instances, the marginal benefit from the incremental cost is too small to be measured. If this viewpoint is correct, and if governments continue to contain medical costs, the future use of technology will be limited to those applications which have a clearly demonstrable marginal benefit associated with their incremental cost. PMID- 2978589 TI - Tissue-derived biomaterials and their use in cardiovascular prosthetic devices. AB - A variety of tissues and tissue components, ranging from allograft and xenograft tissues to albumin- and collagen-coated synthetic materials, have been used to fabricate cardiovascular prosthetic devices. Tissue-derived biomaterials include both viable and non-viable tissues as well as individual tissue components which have undergone some degree of preimplantation processing. A review of the biochemistry, immunogenicity, mechanical properties, physicochemical properties, preimplantation processing and the morphology of the following cardiovascular prostheses are discussed: heart valve bioprostheses and allografts; blood vessel allografts; biologic vascular grafts; and, protein-coated vascular prostheses. This review focuses on the generic aspects of prosthetic device fabrication using tissue-derived biomaterials, realizing that specific differences between various commercially available prostheses may exist. PMID- 2978590 TI - The challenge of small diameter vascular grafts. AB - Symptomatic vascular disease involving blood vessels smaller than 6 mm internal diameter constitutes the majority of vascular disease cases. In many cases the only correction for the condition is vascular reconstruction, or bypass surgery. The only adequate replacement for these vessels at the present time are autologous arteries, or more commonly veins. The availability of a reliable small diameter artificial graft would greatly increase the number of patients that could be treated surgically. Three materials are used for present grafts, tanned natural vessels, and artificial vessels constructed of knitted Dacron or expanded Teflon. All three types eventually fail due to thrombosis, either because of their inherent thrombogenicity or because of encroachment of tissue (intimal hyperplasia) (IH) into the lumen of the graft at the point where the natural and prosthetic vessel join. The exact reasons for thrombosis and IH are poorly understood and constitute a major area of current research. Attempts to alleviate the problem involve: designing non-thrombogenic grafts, lining grafts with the living cells (endothelial cells) that form the inner surface of natural vessels, designing grafts that are elastic like natural vessels, providing biodegradable grafts that are replaced by tissue that reconstitutes a natural vessel, and producing living vessels in vitro. An overview of the problems and current research reveals some promising approaches to the solution, but all are still in the developmental stages. PMID- 2978591 TI - Holographic velocimetry: an alternative approach for flow characterization of prosthetic heart valves. AB - In vitro techniques for evaluating circulation through prosthetic heart valves can yield important information concerning valve performance prior to implantation. Laser Doppler velocimetry is the most widely accepted noninvasive flow measurement technique, however, its use is somewhat limited for flow mapping applications. Holographic velocimetry is an alternative noninvasive in vitro approach for flow diagnostics. The advantage of this technique is the ability to do instantaneous full volume measurements for complete three-dimensional flow mapping at any instant in a flow cycle. Holographic velocimetry was used for the evaluation of circulation through a ball and cage valve. Results indicate that just beyond the apex of the cage very low velocity exists in the mean flow direction, while the other two components of velocity are rather large. Studies done further downstream from the valve begin to show the flow characteristics returning to the state of fully developed turbulent flow, as was previously seen upstream from the valve housing. PMID- 2978592 TI - Fracture mechanics principles applied to implant medical devices--a review. AB - Historically, the fatigue behavior of materials and structures, including biomaterials and implant devices, was analyzed using stress-life (S-N) techniques, i.e. by determining the number of cycles to failure at a calculated peak stress. The development of fracture mechanics technology has made the crack growth aspect of fatigue analysis a more exact science, providing more precise data on the properties of materials, incorporating more geometric and metallurgical information from the structure, and, in some cases, providing for updated in-service lifetime predictions. Over the past decade, the medical implant industry and researchers have adopted this analytical tool to varying degrees. The result has been the development of some improved materials, operative procedures, and devices, based on a more exact knowledge of the properties most critical to assuring the long-term durability of devices. Examples of the application of fracture mechanics technology to orthopedic and cardiovascular devices and the potential for further applications are reviewed. PMID- 2978593 TI - Infections from biomaterials and implants: a race for the surface. AB - Microorganisms in nature and disease are dependent on substratum attachment for optimal growth and development. Similarly, implanted biomaterials tend to potentiate bacteria on their surfaces so that normally friendly special or opportunistic organisms become virulent pathogens. Virulence is also enhanced because both bacteria and biomaterials interfere with host defense mechanisms. Infections centered on biomaterials are most difficult to eliminate and usually require removal of the device. The consequences of device failure are catastrophic and costly. It is the specific nature of the biomaterial surface, which is indirectly a reflection of bulk features, that causes and directs the changes in bacterial behavior which result in virulence. Features of organisms and materials and interactions responsible for these phenomena are reviewed. PMID- 2978594 TI - [Providing antibacterial properties to a vascular prosthesis made of polyethylene terephthalate fibers]. AB - We made the synthesis of a new antibacterial product satisfying the requirements and being similar to polyethylene terephthalate fibres PET from which implants designed for people are made. Proper solvents were worked out and the parameters of processing were determined. The vascular prostheses subjected to the antibacterial processing proved very good and longlasting antibacterial properties. PMID- 2978595 TI - [Problems of conservative dental treatment of patients with long-term dialysis treatment and after transplantation of the kidneys]. PMID- 2978596 TI - [Dental treatment of patients before and after transplantation of the heart]. PMID- 2978597 TI - Evaluation of a new glass ionomer cement with silver as a core buildup under a cast restoration. PMID- 2978598 TI - A simple self-retaining mouth appliance for physically disabled individuals. PMID- 2978599 TI - [Comparison of the causes of occupational atopic dermatitis diagnosed at the Institute of Occupational Medicine in Lodz 1972-1978 and 1979-1987]. AB - The causes leading to occupational contact eczema were compared in two time periods 1972-1978 and 1979-1987. An increase was observed in the second period in the frequency of occupational eczema in women and greater incidence in health service workers. Occupational contact eczema was diagnosed most frequently in construction workers, in machinery and metallurgic industry and in weavers and textile workers. The causes of occupational eczema remained unchanged, the most frequent causes were still hypersensitivity to chromium, cobalt and epoxy resins. No increase in the frequency of hypersensitivity to nickel was noted. PMID- 2978600 TI - [New possibilities of prevention of occupational Candida albicans infection]. AB - The effects were studied in vitro of a score of various additives to toilet soaps on strains of Candida albicans isolated from patients. Their effect on the basic properties of toilet soaps was studied, determining foam production and the index of foam stability, and their resistance to high temperature was determined since in the process of production high temperature might reduce their effects on C. albicans. The strongest fungicidal effect was exerted in strongly alkaline soap solutions by Clotrimazole. Good effects were obtained also using Clioquinol, a derivative of 8-hydroxyquinoline. The results of these studies suggest that certain substances may be added for exerting a fungicidal effect in alkaline medium protecting thus against fungal infections. PMID- 2978601 TI - [Papillon-Lefevre syndrome. Apropos of a case]. PMID- 2978602 TI - [Professional dental practice and AIDS]. PMID- 2978603 TI - [Some aspects of dental treatment for the epileptic child]. PMID- 2978604 TI - An EMG analysis on regulation of the diaphragm and abdominal muscles. AB - Neural mechanisms regulating activities of diaphragm and abdominal muscles were investigated on anesthetized, tracheostomized and spontaneously breathing dogs. EMGs of costal diaphragm and external oblique (EO)abdominal muscle were recorded by fine-wire electrodes. The electrodes for diaphragm-EMG were implanted without opening abdominal cavity using laparoscopy. During quiet breathing the mean firing frequency of diaphragm-EMG was 7.6 +/- 0.7 (S.D.) Hz. The discharge spikes of diaphragm-EMG observed in early-inspiration or in post-inspiratory phase, were low in amplitudes, consistent with previous reports that phrenic motoneurons were comprised of two populations. During quiet breathing spikes were not observed in EO-EMG in a half of the animals, and were tonically-firing in the remaining dogs. Hypercapnia increased firing frequency of diaphragm-EMG, but did not recruit spikes of high amplitude. On the other hand, hypercapnia recruited EO-spikes of higher amplitude, and increased firing frequency. When spikes fired tonically in EO-EMG, hypercapnia suppressed these spikes during inspiration. Airway occlusion gradually recruited EO units of higher amplitude. Mechanical stimulation of upper airway suppressed tonic-EO activity during inspiration. In conclusion, we demonstrated that 1) abdominal wall is not related with control mechanism of diaphragm; 2) the control mechanism of dog's EO differs from cat's internal intercostal muscles; and 3) the tonic component of EO is affected by respiratory commands from higher neuronal architecture. PMID- 2978605 TI - The connective tissue and contractile response of the myocardium to the development and regression of hypertrophy. PMID- 2978606 TI - Validity of endoscopy in the diagnostics of biliary atresia. PMID- 2978607 TI - [The medical emergency in the dental office]. PMID- 2978609 TI - [Growth history and dental treatment of a violent, emotionally-disturbed child]. PMID- 2978608 TI - [Bacteriological examination of dental plaque in institutionalized mentally retarded persons, particularly emphasizing the occurrence of mutans streptococci]. PMID- 2978610 TI - [Bacteriological examination of fissure plaques removed from seriously mentally retarded patients institutionalized]. PMID- 2978611 TI - [Clinical approach to the diseases of psychosomatic dentistry in our department]. PMID- 2978612 TI - [Observations on the medicolegal compensation of the stochastic effects provoked by exposure to ionizing radiation at the work site]. PMID- 2978613 TI - Immunological abnormalities in patient with IgA nephropathy. AB - T cell immunity and phagocytic activity were studied in the blood of patients with IgA nephropathy in order to clarify their roles in the pathogenesis of IgA nephropathy. The percentages of total T lymphocytes, helper T cell and suppressor T cells were significantly reduced in patients. A significantly elevated helper T cell/suppressor T cell ratio in patients showed a predominant reduction in suppressor T cells. There was a significant relationship between histologic findings and helper T cell/suppressor T cell ratio in patients. Natural Killer (NK) cell activity was significantly reduced but the lymphocyte response after phytohemagglutinin (PHA) stimulation was not in patients. ConA-induced suppressor cell activity was not depressed despite of a decrease in suppressor T cells in patients. Phagocytic activity of polymorphonuclear leucocytes (PMNs) ingesting yeasts was significantly reduced in patients. Also an inverse correlation was found between serum IgA levels and phagocytic activity of PMN. It is concluded that suppressor T cell defects, depressed phagocytic activity and impaired NK cell activity may play a role in the pathogenesis of IgA nephropathy. PMID- 2978615 TI - [Dental treatment of handicapped children under general anesthesia at a general hospital]. PMID- 2978614 TI - Familial benign copper deficiency: an old case re-examined. AB - Seven years follow-up and re-examinations of a boy and his mother with "familial benign copper deficiency" revealed repeatedly subnormal serum copper and normal caeruloplasmin levels with relatively good psychosomatic development in the 9 year-old index patient. 64Cu-uptake was elevated in the mother's and normal in the proband's fibroblasts. The pathomechanism of the condition remained unknown, but the biochemical findings and the clinical course did not correspond to any of the previously described forms of congenital defects of copper metabolism. PMID- 2978616 TI - Human delta T cell receptor: rearrangement, deletion, and translocation. AB - Individual T cells express the CD3 molecule in association with alternative gamma delta or alpha beta heterodimeric T cell receptors (TCR). The delta TCR, used in one type of T cell is located within the alpha TCR gene used in quite another cell. T cell precursors and occasional gamma delta T cells in humans possess an unexpected 2.0 Kb mRNA in which a tandemly repeated motif, T early alpha (TEA), has been spliced to the constant (C alpha) region. Long range pulse field gel mapping as well as molecular cloning reveal that TEA is located immediately 5' to the most upstream joining (J alpha) segments of the alpha TCR locus. The human delta TCR locus is immediately 5' to TEA and diversity (D delta 1 +2), J delta 1 +2, C delta, and TEA are linked within 35 Kb. The human delta TCR locus conserves a 12/23 bp spacer paradigm and D delta 1 and D delta 2 are frequently recombined as D delta 1/D delta 2, and reveal exonucleolytic trimming with extensive "N" segment addition. Thus, despite the predominant use of one V delta and J delta segment considerable delta diversity is generated. T cell acute lymphoblastic leukemias (ALL) represent clonal expansions of maturationally arrested cells at specific stages of thymic ontogeny. The gamma delta T-ALLs display allelic exclusion for the delta TCR. Moreover, pre T cells within this group indicate that delta TCR rearrangement can occur prior to the activation of gamma, beta, and alpha loci.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978617 TI - Nakahara memorial lecture. Regulation of T lymphocyte responses: interactions among receptors. AB - Murine helper T lymphocytes (HTL) have been categorized on the basis of the lymphokines they secrete. TH1 cells produce interleukin 2 (IL-2), interferon gamma (IFN-gamma), and lymphotoxin, whereas TH2 cells produce IL-4 and IL-5 but not IL-2 or IFN-gamma. Both cell types apparently can produce IL-3, granulocyte macrophage colony stimulating factor (GM-CSF), and tumor necrosis factor. We have found that there is at least one additional subset which secretes IL-4 and IFN gamma. These helper T lymphocyte subsets also respond differently to immunoregulatory processes. T cell clones of all subtypes can proliferate in response to exogenous IL-2, but IL-4 apparently induces a proliferative response only in those subsets that produce IL-4. IFN-gamma inhibits the proliferation of TH2 clones, but does not affect the proliferation of TH1 clones, cytolytic T lymphocyte (CTL) clones, or cells that produce both IL-4 and IFN-gamma. TH1 cells pretreated with a saturating concentration of IL-2 do not proliferate when stimulated with anti-CD3 monoclonal antibody (mAb). However, proliferation of TH2 cells is enhanced by IL-2 pretreatment, while the TH1 clones or CTL clones through the T cell receptor (TCR) profoundly inhibits IL-2-induced proliferation of those cells, whereas such stimulation either has very little effect or augments the proliferation of IL-4-producing clones. Collectively, these data suggest that the combined influence of these cytokines and intensity of TCR stimulation may determine which cell types expand in number during a particular immunological situation. PMID- 2978618 TI - In vivo administration of anti-CD3 monoclonal antibody can activate immune responses thus preventing malignant tumor growth. AB - Anti-CD3 monoclonal antibodies (mAb) have been shown to suppress T cell-mediated immune responses both in vitro and in vivo. However, in vitro studies with these antibodies have also demonstrated that they possess potent mitogenic properties, raising the possibility that they might be capable of potentiating immune responses in vivo. In this regard, we have recently shown that an anti-CD3 mAb can activate murine T cells in vivo. Furthermore, low doses of antibody induce interleukin 2 (IL-2) receptor expression and enhanced proliferation to allogeneic major histocompatibility complex (MHC) antigen without detectable modulation or blocking of the T cell receptor and without suppression of T cell-mediated immune responses. In light of these findings, we investigated the ability of low dose anti-CD3 to enhance an anti-tumor response directed against the malignant murine UV-induced skin tumor, 1591-Pro-4L. Low dose anti-CD3 administration resulted in enhanced in vitro anti-tumor activity and prevented tumor outgrowth in approximately two-thirds of animals treated at the time of tumor inoculation. Furthermore, these animals displayed lasting tumor-specific immunity. These results suggest that anti-CD3 mAb can be utilized for the enhancement of anti tumor responses in vivo and may have general application in the treatment of immunodeficiency. PMID- 2978619 TI - Analysis of melanoma antigen and its involvement in tumor-escape mechanisms. AB - Melanoma antigen was characterized by using the C57BL/6 mouse melanoma (B16) system, especially in relation to escape mechanisms of tumor cells from immunological surveillance. The antigen on the surface of melanoma cells selectively induced double negative cytotoxic T lymphocytes (CTL) lacking genetic restriction specificity in their action, whereas the soluble antigen shed or secreted from the cells preferentially induced suppressor T cells (Ts) inhibiting CTL generation in the induction phase. The epitopes of melanoma antigen for CTL and Ts were found to possess a "GM3-like structure". Anti-melanoma CTL activity was blocked by either GM3(NeuAc)-or GM3(NeuGc)-liposomes. Moreover, the GM3 (NeuGc)-liposome could induce anti-melanoma CTL when used as an antigen in the in vitro primary response. On the other hand, the soluble melanoma antigen or GM3(NeuAc)-but not GM3(NeuGc)-liposome itself specifically induced anti-melanoma Ts. Therefore, anti-melanoma Ts are able to distinguish GM3 molecular species. We also found two types of T cells, C3T4+ and double negative I-J+ T cells, to be involved in this suppression. Although the primary structure of melanoma GM3 was demonstrated to be the same as that of normal GM3, syngeneic anti-melanoma GM3 monoclonal antibody (M2590) did distinguish melanoma from normal cells. Further close analysis in liposome lysis experiments using various concentrations of GM3 clearly demonstrated that M2590 anti-melanoma GM3 only reacted with GM3 at a "high" density (more than 10-12 mol%), whereas no reactivity was observed at a "low" density (less than 7.5 mol%). It is clear, therefore, that the density of GM3 with normal primary structure is important in generating melanoma antigenicity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978620 TI - IL-2 receptor and Fc epsilon R2 gene activation in lymphocyte transformation: possible roles of ATL-derived factor. AB - Lymphocyte transformation/activation is accompanied by the induction of a variety of the inducible receptors associated with the activation antigens. The p55 chain of the interleukin-2 receptor (IL-2R/p55) recognized by anti-Tac monoclonal antibody (mAb) is expressed on activated T and B cells as well as natural killer (NK) cells. IL-2R/p55(Tac) is constitutively expressed on T4(+) T cells transformed by human T-lymphotropic virus I (HTLV-I), which is a causative agent for adult T-cell leukemia (ATL). Low affinity Fc receptor for IgE (Fc epsilon R2/CD23) is another inducible receptor binding IgE and is expressed on various hematopoietic cell types. While the physiological expression of Fc epsilon R2 and its soluble form (IgE Binding Factor; IgE BF) is variably regulated by cytokines and IgE, there is a constitutive expression of Fc epsilon R2 on Epstein-Barr virus (EBV) transformed B cell lines and some of HTLV-I (+) T cell lines. ATL derived factor (ADF) has been characterized as an IL-2R/p55(Tac) inducing factor derived from HTLV-I(+) T cell lines. Purification of ADF protein and the cDNA cloning proved that ADF is closely related to the autocrine growth factor produced by an EBV(+) B cell line 3B6 (H. Wakasugi and T. Tursz) and belonging to the family of thiol reducing co-enzyme thioredoxin which is involved in many biological reactions. Recombinant ADF produced by COS cells and E. coli showed both IL-2R inducing and reducing activities. We will discuss the biological roles of ADF in viral and normal lymphocyte activation and transformation in relation to the receptor gene activation. PMID- 2978621 TI - The T-cell antigen receptor: a complex signal-transducing molecule. AB - The T cell antigen-specific receptor (TCR) on most mature T cells is a multisubunit complex composed of 7 chains: alpha, beta, gamma, delta, epsilon, and either a zeta-zeta homodimer (zeta 2) or a zeta-eta heterodimer (zeta eta). We have derived a series of TCR variants and mutants from the antigen-specific murine T cell hybridoma, 2B4.11, permitting detailed analyses of the assembly and transport of the TCR. Loss of the zeta chain resulted in markedly reduced cell surface TCR expression. This was due to enhanced degradation of the other TCR chains in a post-Golgi, probably lysosomal, compartment. Loss of the beta chain, delta chain, or the combination of delta and zeta chains, also resulted in loss of cell surface TCR expression. Unlike the zeta loss variants, in these cases the other TCR chains were retained in the ER. Epsilon, gamma, and zeta could survive for prolonged periods in the ER, while the alpha, beta, and delta chains were rapidly and efficiently degraded. In another series of studies, variants that were deficient in the eta chain but that expressed normal levels of zeta 2 containing TCRs were analyzed for their functional properties. A positive relationship was found between the presence of zeta eta and the ability to respond to mitogenic stimuli with increases in phosphoinositide hydrolysis and, in one well characterized variant, increases in intracellular Ca2+. Despite this, late biological responses such as interleukin 2 (IL-2) production and inhibition of transformed growth were relatively normal. These results call into question the putative cause-and-effect relationship between some early biochemical events and these late biological responses. Further, they suggest a model in which zeta eta-containing TCR complexes are largely responsible for activation-induced phosphoinositide hydrolysis. PMID- 2978622 TI - [Microleakage between light-cured composite resins]. PMID- 2978623 TI - [Marginal leakage test on "Silux" composite resin]. PMID- 2978624 TI - Periodontal probing and its relation to degree of inflammation and bleeding tendency. PMID- 2978625 TI - Cuspal reinforcement in primary teeth: an in vitro comparison of three restorative materials. PMID- 2978626 TI - Psychological factors in the treatment of chronic low back pain. Follow-up study of a back school intervention. AB - The aim of the study was to investigate the role of certain psychological factors (e.g. neurotic features, alexithymia, and hostility) as intervening variables modifying the outcome of the back school intervention or correlating with spontaneous recovery. The results indicated that those patients who reacted favorably to the back school intervention could be described as emotionally well adjusted and controlled showing relatively good cognitive capacity with undisturbed reality testing. The poor responders in the treatment group were less capable cognitively and not so well balanced emotionally. Patients showing spontaneous recovery in the control group were characterized by a more lively and less controlled way of expressing emotions and affects. In contrast to these, patients who showed increasing disability during the 1-year follow-up were characterized by restricted expression of emotions and affects indicating alexithymia. PMID- 2978627 TI - Glass ionomer-silver cermet bonded composite resin Class II tunnel restorations. PMID- 2978628 TI - Glass ionomer-silver restorations: a demineralization-remineralization concept. PMID- 2978630 TI - [Important aspects of the development of a syndrome. A dental viewpoint]. PMID- 2978629 TI - [Conservative dentistry and risk of Osler's disease]. PMID- 2978632 TI - [A histological study on the changes of the lingual papillae of streptozotocin induced diabetes in rats]. PMID- 2978631 TI - [An electron microscopic study on the capillaries of buccal mucosa and retina in streptozotocin-induced diabetic rats]. PMID- 2978633 TI - [Desmoid tumors]. PMID- 2978634 TI - Atrial natriuretic factor in the impulse conduction system of the heart. PMID- 2978635 TI - Chronic infusion of atrial natriuretic peptide prevents pulmonary hypertension in hypoxia-adapted rats. PMID- 2978636 TI - Intrarenal dopamine-1 receptors control renal function. AB - Dopamine receptors are classified to DA-1 and DA-2 and are characterized in renal tissue by radioligand binding and by the response of renal adenylate cyclase to dopaminergic agonists and antagonists. DA-1 receptors are localized in the renal tubules, the medial layer of renal microvessels, and the juxtaglomerular apparatus. DA-1 receptor stimulation causes dilation of renal, mesenteric, coronary, and cerebral vessels. In the present study, we tested the hypothesis that dopamine is a paracrine substance in the control of renal function. We employed a potent specific DA-1 receptor antagonist, SCH, to evaluate the role of intrarenal DA-1 receptor in the maintenance of renal function. Intrarenal DA-1 receptor blockade with SCH caused a highly significant dose-dependent antidiuresis and antinatriuresis, and decreased FENa. A rebound diuresis and natriuresis above control values were observed after cessation of DA-1 receptor blockade. There were no changes in renal hemodynamic function during DA-1 receptor blockade. These results strongly suggest that the antinatriuresis and antidiuresis induced by DA-1 receptor blockade are mediated by an action at the renal tubule. The infusion rate of SCH administered intrarenally was sufficiently low to produce no measurable systemic effects including PRA, PAC, and MAP. Thus, these results can be interpreted as due to intrarenal DA-1 blockade. In summary, we have demonstrated that renal excretory function is highly sensitive to DA-1 receptor blockade within the kidney and appears to be mediated by renal tubular events. This study provides strong evidence that DA-1 receptors play a physiological role in the control of renal function. PMID- 2978637 TI - Palmoplantar keratoderma with normal intelligence in tyrosinaemia II. PMID- 2978638 TI - Hypereosinophilic syndrome presenting with erythroderma. PMID- 2978640 TI - [Hyperplastic gingival conditions from calcium antagonists]. PMID- 2978639 TI - [How to treat heart patients]. PMID- 2978641 TI - Cleft lip and palate anomaly, retinitis pigmentosa, epilepsy and twinning. PMID- 2978642 TI - [Immediate preprosthetic restorations for nonvital teeth]. PMID- 2978643 TI - [Extractions in hemophilic patients. Control of hemostasis with DDAVP]. PMID- 2978645 TI - AIDS and infection control. PMID- 2978644 TI - Coping behaviour of disabled persons and their families: cross-cultural perspectives from Norway and Botswana. AB - In this article a model for analyzing the coping behaviour of families with disabled children is presented. It is demonstrated how this model, through its holistic perspective, may be used for including culture as an important variable in studies of reactions to crisis and coping behaviour in such families. Some cross-cultural perspectives, based on studies from Norway and Botswana are presented, and the universality of reactions to critical events, as established in prominent theories of crisis are questioned. PMID- 2978646 TI - Enid dentists help the forgotten. PMID- 2978647 TI - Isolation and properties of rat submandibular gland N-acetylhexosaminidase. PMID- 2978648 TI - Treatment of the infrabony defect: elimination or regeneration. PMID- 2978649 TI - Biochemical approach to periodontal regeneration--a review of the literature. PMID- 2978650 TI - Dentists called to account over HIV infected patients. PMID- 2978651 TI - [Nutrition in the elderly]. PMID- 2978652 TI - Growing pains. PMID- 2978654 TI - [The attitude toward treatment of HIV-positive patients has improved]. PMID- 2978653 TI - Dentistry: the law and mental retardation. PMID- 2978655 TI - [Development of dental care for disabled patients]. PMID- 2978656 TI - [Immunization against dental caries]. PMID- 2978657 TI - Dentists split on AIDS. PMID- 2978658 TI - The ethical and legal considerations of treating patients with infectious diseases. Canadian Dental Association. PMID- 2978659 TI - Woodall's editorial on AIDS excellent. PMID- 2978660 TI - [Therapeutic catheterization in cardiology of adults]. PMID- 2978661 TI - [The human immunodeficiency virus and transmissible infections. Analysis of the problem and recommendations for dental care]. PMID- 2978662 TI - [Hemodynamic effects and plasma catecholamine concentrations during dental treatment in hypertensive patients]. PMID- 2978663 TI - [Histopathology and microradiography of changes in rat tibia epiphyseal cartilage after streptozotocin administration]. PMID- 2978664 TI - An unusual hazard. PMID- 2978665 TI - Effects of low temperature on the respiratory metabolism of carbohydrates by plants. AB - The effects of lowering the temperature from 25 degrees C to 2-8 degrees C on carbohydrate metabolism by plant cells are considered. Particular emphasis is placed on the mechanism of cold-induced sweetening in tubers of potato (Solanum tuberosum). Temperatures between 0 and 10 degrees C were shown to cause a marked reduction in the rate of respiration of a wide range of plant tissues. At these temperatures the ability of suspension cultures of soybean (Glycine max), and callus cultures and tubers of potato to metabolize [14C]glucose was appreciably diminished. The detailed distribution of 14C showed that lowering the temperature decreased the proportion of the metabolized [14C]glucose that entered the respiratory pathways and increased the proportion converted to sucrose. Pulse and chase experiments, in which [14C]glucose was supplied to potato tubers at 2 and 25 degrees C, showed that lowering the temperature led to accumulation of label in hexose 6-phosphates, which were subsequently converted to sucrose. The patterns of 14CO2 production from specifically labelled [14C]glucose supplied to soybean suspension cultures and disks of potato tuber suggested that lowering the temperature reduced the activity of glycolysis more than that of the oxidative pentose phosphate pathway. It is argued that the above experiments demonstrate that lowering the temperature not only reduces the rate of carbohydrate metabolism but also alters the relative activities of the different pathways involved. A disproportionate reduction in glycolysis at the lower temperatures is suggested. Mature tubers of many varieties of potato accumulate sucrose and hexose when stored between 2 and 10 degrees C. Starch is the source of carbon for this synthesis of sugar. We could not detect cytosolic fructose-1,6 bisphosphatase in potato tubers and suggest that carbon for sugar synthesis in the cold leaves the amyloplast, not as triose phosphate, but probably as a six carbon compound. Evidence is presented that phosphofructokinase (EC 2.7.1.11) plays a major role in regulating the entry of hexose 6-phosphates into glycolysis in potato tubers. Phosphofructokinase was purified from potato tubers and shown to consist of four forms. Three of these forms were shown to have higher Q10 values over the range 2-6 degrees C than over the range 12-16 degrees C and are regarded as being cold-labile. No such cold-lability was detected for the key enzymes involved in sucrose synthesis and the oxidative pentose phosphate pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978666 TI - [Dental care in Danish psychiatric institutions]. PMID- 2978667 TI - [The general physical, psychological and social situation among patients with cerebral palsy]. PMID- 2978668 TI - Plastic surgery of the trunk--reducing blood loss. AB - A total of 178 patients has been reviewed who underwent major plastic surgery over a 6-year period. This series compares well with other similar series. No transfusions were required and the average blood loss was estimated at 150 cm3. Dilute epinephrine infiltration and coagulation cautery dissection effectively controls and reduces blood loss in major plastic surgery of the trunk. PMID- 2978669 TI - An abdominal repair for complete rectal prolapse. AB - A modified sling rectopexy to the sacrum was performed in a series of 104 patients with complete rectal prolapse during 1975-86. A sling of mersilene mesh was sutured to the front of the sacrum and to the sides of the rectum without enclosing its anterior surface. A postoperative mortality of 2% and morbidity of 23% were found. Control of the prolapse was achieved in over 90% of cases and the continence rate rose from a preoperative level of 37% to 86% postoperatively. This form of abdominal repair is an effective method of correcting complete rectal prolapse in the majority of cases. PMID- 2978670 TI - [Oral complications in patients with acute leukemia]. PMID- 2978671 TI - [Dental management of children with cerebral palsy]. PMID- 2978672 TI - [Basic standards for dental care of patients with AIDS]. PMID- 2978673 TI - Digital signal processing applied to Doppler U.S. velocimetry. PMID- 2978675 TI - Unexpected drug reactions. PMID- 2978676 TI - Cutaneous drug reactions in porphyrias. PMID- 2978674 TI - A brief history of drug reactions. PMID- 2978678 TI - Adverse reactions to antibiotics other than penicillin. PMID- 2978677 TI - Adverse reactions to penicillin and related drugs. PMID- 2978679 TI - Identifying adverse reactions to drugs. Reporting systems. PMID- 2978680 TI - Adverse effects of corticosteroids: I. Topical and intralesional. PMID- 2978681 TI - Adverse effects of corticosteroids: II. Systemic. PMID- 2978682 TI - Lithium, leukocytes, and lesions. PMID- 2978683 TI - Medical complications of isotretinoin. PMID- 2978684 TI - Cutaneous drug reactions in small animals. PMID- 2978686 TI - Immunology of adverse drug eruptions. PMID- 2978685 TI - Histology of adverse cutaneous drug reactions. PMID- 2978687 TI - Systemic contact-type dermatitis due to drugs. PMID- 2978689 TI - Left ventricular hypertrophy in hypertension and the effects of antihypertensive drug therapy. PMID- 2978688 TI - Drug reactions involving the mouth. PMID- 2978690 TI - Finishing of polymaleic/polyacrylic based glass polyalkenoate cements. PMID- 2978691 TI - [Antibiotic prophylaxis for patients at risk in dental practice]. PMID- 2978692 TI - [Impairment presuppositions in the application of the Article No. 5 of the Law No. 222 of 6/12/1984 (allowance for attending or nursing)]. PMID- 2978693 TI - [Ketac silver: in vitro experience and clinical results]. PMID- 2978694 TI - Shear bond strength of Scotchbond 2/Silux to dentin. PMID- 2978695 TI - Coping with disability. PMID- 2978696 TI - [Mechanisms of action of endo- and exoactive substances in psoriatic skin]. PMID- 2978697 TI - [Reversible functional changes in the heart and circulation in extreme athletic exertion]. PMID- 2978698 TI - Special dental clinic at UMDNJ to serve AIDS, other patients. PMID- 2978699 TI - AIDS in perspective. PMID- 2978700 TI - [Echocardiography in primary hemochromatosis]. PMID- 2978701 TI - [Psychogeriatrics: tentative evaluation of mental health in the aged]. PMID- 2978702 TI - [Surgical treatment of teeth in hemophiliacs]. PMID- 2978703 TI - [Genetic deficiencies of beta-N-acetylhexosaminidase A and B: Tay-Sachs and Sandhoff diseases (GM2-gangliosidosis)]. PMID- 2978704 TI - [Aspects of prophylactic orientation in stomatology]. AB - The prophylactic orientation is a basic principle, the application of which, in the stomatological practice, on a constant basis, aims at a significant improvement in the status of bucco-dental health of the population, and the transformation of the present type of stomatology, in a truly modern stomatological medicine. The author makes a detailed analysis of the strategy employed in the reduction of the provence of the main bucco dental diseases, and of the necessities for stomatological treatment, especially of the reduction by over 50% of the prevalence of dental caries, which, at the present time, makes necessary to carry out approximately 75% of the entire volume of stomatologic activities in adults. Also the reduction is attempted, of the prevalence of gingivitis, parodontitis, dento-maxillary anomalies, of the cancer of the buccal cavity, and other affections, by methods and means of primary prophylaxis, applied in the majority of the population. The secondary prophylaxis is also analysed, as it is carried out by early detection and treatment of lesions and diseases in the dento-maxillo-facial field. Other aspects are also discussed, such as: extension of the prophylactic orientation in the curative stomatological practice, and the avoidance of iatrogenic lesions and diseases, which are frequent in the stomatological practice; the ergonomic organization of the working place, and of the work methods, reduction of the risks involved in contracting or transmitting to or from the patients of various diseases, prophylaxis of occupational diseases in stomatologists, prophylaxis of aspects, attitudes or behaviour which are against etichal and deontological principles of the profession, as well as the necessity for a better preparation from the prophylactic viewpoint of stomatologists and their coworkers in view of coping with increased competence and efficiency with the increased demands of stomatological assistance of the population. PMID- 2978706 TI - [Pharmaceutical preparations from plant products employed in stomatologic diseases]. AB - Two preparations for stomatological use have been introduced, obtained from vegetal extracts of Ocimum basilicum L., Tilia sp., and Symphytum officinalis L. Their stability in time was evaluated, and the microbiological activity was assessed in vitro on microbial strains involved in stomatological affections. In clinical experiments good results were obtained in parodonthopathies, in the therapy of lesions developing under prostheses, and in other affections. PMID- 2978705 TI - [Juvenile periodontosis. Note 1. Clinical and morphopathologic considerations]. AB - The authors review a series of aspects related to the classification and the terminology of parodonthopathies. They propose to investigate some clinical and morphopathological aspects in 14 cases of juvenile parodonthosis, a form of parodonthopathy that has been accepted since 1971 (Baer). The characteristic clinical aspects are stressed, of this affection, including: the "mirror-like" aspect of the lesions, involvement of the frontal and lateral group of teeth, and the 6-year molar--a mode especially seen in women, as well as the histopathologic characteristic changes which accompany this affection. Thus, in parallel with the dystrophic processes, and the degenerative ones, there are also characteristic signs of chronic inflammation in the parodonthal membrane. PMID- 2978707 TI - [Actualities concerning frequency, etiopathogeny, and the diagnosis of temporomandibular osteoarthrosis]. AB - On the basis of a vast reference list, and on the personal experience of the authors new opinions are presented, concerning the concept, the frequency, and the etiopathogeny of temporomandibular osteoarthrosis. Although the symptoms and signs of this morbid entity are frequently similar with those of other intracapsular affections, as a result of a combination of modern radiographic investigations (arthrotomography with double contrast, computerized scanner tomodensitometry, magnetic nuclear resonance), arthroscopy, macroscopic and microscopic examinations (biochemical analysis of the synovial fluid) it has become possible to make a positive diagnosis, as well as a differential one. The conclusions of the study have confirmed the evolution of a large number of experimental studies, both clinical and epidemiological, published in the specialized literature, which favor the detection and the removal of multiple risk factors involved in the development of this disease of degenerative origin, by a correct treatment, and by the use of prostheses. PMID- 2978708 TI - [A procedure for improving the stability of the total mandibular prosthesis with the aid of the muscular factor in the marked atrophies of the mandibular crest]. AB - The study aimed to define factors involved in improving the stability of total mandibular prostheses by the use in optimal conditions of the muscular factor. This is achieved with a special method for casting of the neutral space which allows to prepare the external surface and to place the artificial dental arc according to objective functional criteria. In the first part of the study notions are discussed related to the practical solution of the problem, such as: the anatomo-functional study of formations which define the neutral space, aspects of the neutral space in the total edentate, and the conformation of total prostheses, as well as the placing of the artificial dental arcs in relation to the neutral space. In the second part a method is presented, for making prints, that was used in 35 cases. The results of the prosthetic procedures, verified clinically and electromyographically are promising. PMID- 2978709 TI - [Optimizing therapeutic methods in mandibular fractures]. AB - The authors present various methods for immobilizing mandibular fractures, stressing their advantages and disadvantages. They describe modern surgical methods used in immobilizing these fractures, considering methods employed in stable, functional osteosynthesis that have improved indications for surgical therapy, and that have made obsolete intermaxillary immobilization. These methods are more acceptable for the patient because they allow for a completely normal diet, as well as the maintenance of a normal hygiene in the buccal cavity. Evolution of the recovery is more easily followed, and in case of complications interventions are easily carried out in a short time. The presence of functional stimuli enhances the development of a good callus, and the full recovery is shortened by 2-3 weeks. PMID- 2978711 TI - [Acupuncture as a bioenergetic reequilibration means in the treatment of chronic mixed marginal parodontopathies]. AB - The authors discuss the use of acupuncture in the treatment of chronic mixed marginal parodontopathies as an eventual means of bioenergetic reequilibration. The various studies carried out for some time by the Department of Odontology and Parodontology of the Bucharest Faculty of Stomatology have demonstrated the lateration of electrical bioresistance in certain acupuncture points in the cephalic extremity at the level of the gingival mucosa in pathologic parodontal states. This suggested the possible applicability in chronic mixed marginal parodontopathies of acupuncture instead of the known means of biostimulation. PMID- 2978710 TI - [Contribution of immediate orthopedic-orthodontic treatment in maxillo-palatine labial clefts]. AB - Labio-maxillo-palatine clefts are pathological conditions accompanied by severe handicaps, congenital malformations of particular interest for medical assistance because of several reasons, including the following: they are the most frequent congenital malformations of the bucco-maxillo-facil region: in certain cases there are other teratologic conditions in other parts of the organism that are associated to these clefts; there are no disturbances from the intellectual viewpoint; there are efficient possibilities for therapy as a result of a multidisciplinary approach, and the progress achieved in each of the fields involved may contribute to continuous improvement of the therapeutical prognosis; because of the therapeutical problems that these pathological conditions may raise, and because of the long duration of the treatment, the pathology involved in labio-maxillo-palatine clefts has a special social character which renders necessary the support of extra-medical factors (special schooling, special conditions for work, etc.). All these aspects explain the world-wide interest for this type of pathology, as it is also reflected in the large number of scientific manifestations dedicated exclusively, or in association to this problem, aimed at achieving a permanent therapeutic progress in this field. In the period between 1986 and 1987 the authors have selected 60 patients out of those hospitalized for this defect in the Clinic. The patients were aged between 2 and 43 years. Orthodontal therapy is the type of therapy which takes the longest time in these patients. This is achieved by the successive application of orthodontal devices, which necessitate a long-term therapeutic strategy. In the course of this therapy a permanent cooperation is necessary between the specialist in orthodontics and the maxillofacial surgeon. A common approach is beneficial for both specialties. The number of specialists in orthodontics involved in the orthodontal therapy of labio-maxillo-palatine clefts is alarmingly low, and thus there is no possibility to evolve a special program for their further special education. Another difficulty is the fact that patients from all over the country demand the services of the same specialist, and a series of aspects evolve from this situation, as follows: the patients (usually accompanied by one of the parents) have to perform long trips, which are also expensive; when incidents occur, in relation with the use of a orthodontal device these patients cannot require immediately the assistance of specialized services, and, as a result, the number of recidives is alarmingly high.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2978712 TI - [Correlation between occluding trauma and dental erosion]. AB - The study presents data obtained by examination of a lot of patients with erosions of the teeth. At the same time histologic sections were prepared from teeth with cuneiform lesions that had to be removed as a result of parodonthopathic processes. Histological studies that have been carried out have demonstrated hyperemia of the dental pulp, and blood suffusion, sometimes accompanied by oedema. Slight rarefaction of dentine was also noted in other microscopic fields, with characteristic enlargement of the interdentinal spaces. The fact that the histological picture does not provide too much data on the etiopathogenic mechanism of the cuneiform lesions it may be considered that the occluding trauma is one of the major causes of these coronal lesions. Occluding balancing as performed by the authors in these patients have always demonstrated the presence of erosions in teeth that had suffered from occluding trauma. By this attitude and with these means of therapy one can prevent the development of new lesions of the cuneiform type, and thus contribute to the decrease in the morbidity, and to the prevention of the various disturbances in the dentomaxillary apparatus. PMID- 2978713 TI - [Edentation of the 6-year molar tooth. Clinical aspects and treatment]. AB - The problem of the 6 year molar tooth edentation has been approached from an unilateral point of view. The present paper deals with the complex factors that contribute to the onset and evolution of the 6 year molar tooth edentation. The group studied included 2000 subjects making it possible to establish the frequency of the affection in terms of age, general condition of the body environmental factors, caries prevention, out-patient follow up, previous dental treatment. The study continued with 100 cases of molar edentation without prostheses, establishing the clinical and complementary signs of the 6 year molar syndrome. Likewise discussed are the favourable elements and errors in the concept and treatment of previously applied prostheses. A complex treatment was applied in 50 cases of 6 year molar edentation showing the preprosthetic, proprosthetic and prosthetic treatment applied and certain therapeutical techniques applicable by the practitioner. PMID- 2978714 TI - [The clinical and technological particularities of using Gaudent "S" and Gaudor alloys in the conjoint treatment of extensive edentation]. AB - The present paper forms part of a longitudinal study on prostheses restaurations with Romanian non-noble alloys for establishing the clinical, technological and biological implications of their utilization in mass assistance. The large contact surface with biological structures and the buccal medium of total conjoint restaurations permitted chacking of previous data regarding the biological behaviour of these alloys. In the group of 58 patients with total bridges followed up for 2 to 5 years there were no adverse reactions in which the alloy used could be incriminated. The iatrogenic aspects observed consequent to the application of prostheses were common to the use of any other metal alloy. The surface colour changes in a reduced number of cases depended upon the technologic preparation of the alloys and particularly upon the patient's limited ability to keep the plate under bacterial control. The study demonstrated that casting of a total Gaudent S bridge under present conditions does not give correct adaptation of the cervical limits which means more accurate compliance with the technology and homologation of the specific of this alloy. PMID- 2978715 TI - [The transverse cervical flap. Indications and results]. AB - The transverse cervical regional flap may be used for covering defects, especially after oncologic exeresis against a potentially reduced biological background in the regions of the chin, parotid gland, the ear and buccal cavity. The authors used this flap in seven cases: 5 carcinomas, an avulsive facial wound and a retractile scar of the chin. There were very good results in 4 cases, good in 2 cases, and in 1 case necrosis of anterior portion of the flap developed. As the results of the use of this flap have not been published we consider the present publication contributes to the clinical indications of the method and its limitation. PMID- 2978716 TI - Is alpha-ANP primarily a natriuretic hormone? PMID- 2978717 TI - Effects of combined hypophysectomy and cyproterone acetate on the adrenal chromaffin cells of the rat. Evidence for postnatal migration of adrenal chromaffin cells. AB - A light and electron-microscopic study was performed concerning the effects of hypophysectomy (HP) followed by cyproterone acetate (CA) treatment on the adrenal chromaffin cells of the rat. The latter drug is reported to interfere with steroid biosynthesis. When given following HP, CA was found to induce degeneration in the inner cells of the zona fasciculata and a marked fall in the blood corticosterone level. The adrenal chromaffin cells which were depleted after the administration of reserpine at the beginning of the experiment failed to recover two weeks later. In addition, some of the depleted chromaffin cells migrated towards the outer cells of the zona fasciculata, had the appearance of pheochromoblasts and showed features of increased synthetic activity. The results are discussed in the light of the generally accepted functional relationship between the cells of the adrenal cortex and the medulla and following a marked fall in the concentration of blood corticosterone, a chemobiotaxis is suggested between the chromaffin and adrenal cortical cells. PMID- 2978719 TI - The absence of non-local conformational changes in OR3 operator DNA on complexing with the cro repressor. AB - The Interaction of the cro protein of lambda phage with a synthetic OR3 operator having 17 base pairs in length and with its 9 bp fragment has been studied using the circular dichroism (CD) method. In both cases, a considerable change in the CD of the samples was found in the region 260-300 nm upon the addition of the cro protein. The stoichiometry obtained by the CD titration was identical for OR3 and its 9 bp fragment: one duplex per dimeric cro. NaCl addition makes the complexes dissociate so that the 9 bp fragment becomes free at [NaCl] greater than 0.2 M while the whole OR3 becomes free at [NaCl] greater than 0.5 M. The CD spectra of both the free duplexes show a typical B-form conservative pattern with a positive CD band (270 nm) and a negative one (250 nm). The specific complexing of both the duplexes results in a substantial CD depression in the positive band. The most pronounced effect occurs at 280 nm. This spectral change is quite distinct from those in the B to A transition and in the non-cooperative winding of the DNA within the B-family of forms. The interaction of the cro protein with the non operator DNAs, calf thymus DNA and a synthetic 10 bp duplex, reveals no visible CD changes at all. An inference is drawn that the CD change in the specific complexes is mainly due to the induced CD in tyr-26 upon its interaction with a specific base pair in the operator or its fragment, the operator DNA conformation being conserved in a B-like form as a whole.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978718 TI - Peculiarities of the B to A transition of the lambda phage regulatory site OR3 and of its fragment. AB - Conformations of the synthetic deoxyoligonucleotide 17 base pairs long, which is an OR3 operator of lambda phage, and of its 9-b.p. fragment were studied by the circular dichroism method (CD). The regions of stability of the double-stranded state were determined for these duplexes. A comparison of the CD spectra for these oligonucleotides with the CD for a lengthy DNA showed the conformation of these short DNA pieces to belong to the B-family. A cooperative change in the CD spectra is observed in trifluoroethanol (TFE) solutions at a TFE concentration specific for each oligonucleotide, which is supposed to stem from a B to A transition. The length of the fragment was found to affect the ability for the B A transition. The transition into the A form is hindered by 13% TFE for the short 9-nucleotide in comparison with the 17-nucleotide. We suggest that this is due to the B form stabilization by terminal base pairs (B-phility of the ends). PMID- 2978720 TI - Sequential radiation damage in protein crystallography. AB - Radiation damage in protein crystals is described in terms of a sequential process of protein disordering. A new radiation-damage model has been tested against data from several protein crystals and can describe radiation damage corresponding to loss of the original intensity in excess of 80%. The model is an extension of previous models which characterize radiation damage in terms of successive conformational transitions of the protein from an undamaged to a spatially disordered to finally an amorphous state. The proposed model provides a more-general positional characterization of the disordered protein and includes, prior to the disordered state, a new dose-dependent state in which the protein conformation resembles the undamaged protein. Comparison of this model with the best previous model shows that the proposed model provides an improved fit to radiation-damage data. PMID- 2978721 TI - Prevention of bacterial endocarditis in localised juvenile periodontitis and Papillon-Lefevre syndrome patients. AB - The bacterium Actinobacillus actinomycetemcomitans is found in large numbers in subgingival plaque and gingival tissues of patients with LJP and PLS. This bacterium too has been found to cause infective bacterial endocarditis in patients at risk. Antibiotic prophylaxis is necessary for at risk patients with LJP and PLS because significant bacteraemia is produced during extensive periodontal instrumentation, extractions and surgery which are required in managing these cases. The current antibiotic regimens recommended by the American Heart Association/Council on Dental Therapeutics are not effective against this bacterium. A two-stage prophylactic approach is advocated, first with tetracycline for two weeks to eliminate the Actinobacillus actinomycetemcomitans, followed by the regimens recommended by the American Heart Association on the day of the dental procedure itself. Tetracycline should not be used concurrently or as a substitute for the recommended regimens by the American Heart Association/Council on Dental Therapeutics. PMID- 2978722 TI - [Distal tubular acidosis. Dental aspects]. PMID- 2978723 TI - New York Court approves AIDS referral. PMID- 2978724 TI - [The use of OsO4-pyridine reagent for studying decarboxylated amino acid complexes with DNA]. PMID- 2978725 TI - [Membrane ATPase and erythrocyte cations in acute leukemia]. PMID- 2978726 TI - [Plasma level of atrial natriuretic peptide in patients with uncomplicated essential hypertension]. PMID- 2978727 TI - Antigenic targets in membranous glomerulonephritis. PMID- 2978728 TI - [Sarcoplasmic reticulum of skeletal muscles and calcium pump]. PMID- 2978729 TI - [The role of phospholamban in cardiac sarcoplasmic reticulum]. PMID- 2978730 TI - [Myosin-linked Ca2(+)-regulation of action-myosin-ATP interaction]. PMID- 2978731 TI - [Urodynamics in normal pediatric patients]. PMID- 2978733 TI - Bacteriophage lambda int protein may recognize structural features of the attachment sites. AB - The bacteriophage lambda int protein binds to and promotes polynucleotide strand exchange within specific DNA segments called attachment sites. Previous work strongly suggests that the specificity of int protein action is based, at least in part, on its ability to recognize nucleotide sequences in the attachment sites. We suggest that int protein also recognizes structural features of the attachment sites such as the twist and roll angles between adjacent base pairs. This proposal is based on statistical analysis of the predicted twist and roll angles of a large collection of secondary attachment sites. The analysis shows that the oscillation patterns of these parameters are conserved in regions where int proteins binds. PMID- 2978732 TI - NMR studies of DNA recognition sequences and their interaction with proteins. The phage lambda OR1 operator, a symmetric lac operator and their specific complexes with cro protein and lac repressor "headpiece". AB - The phage lambda operator OR1 and a 18 base pair symmetric lac operator have been studied by high resolution NMR. The imino proton resonances and the resonances of the unexchangeable protons (except the 5' and 5" sugar proton resonances) have been assigned by one- and two-dimensional NOE techniques. The imino proton resonances of OR1 and the symmetric lac operator have been used to monitor changes induced in the operator structure by the formation of a specific complex with the phage lambda cro protein and with the lac repressor N-terminal DNA binding domain ("headpiece"). Two regions within the OR1 sequence could be identified, where changes in the imino proton resonance positions occur: The central part around base pairs CG 9 and 10 and the region around base pairs AT 5 and CG 5. The TA base pair 6 is the only position in the symmetric lac operator, where the complex formation with headpiece induces a change. PMID- 2978734 TI - A study of phi X-174 DNA torus and lambda DNA torus tertiary structure and the implications for DNA self-assembly. AB - Hydrated torus shaped complexes were examined by transmission electron microscopy in both spermidine-condensed linear and nicked circular phi X-174 DNA and lambda DNA preparations. Freeze-etch replicas of both these torus samples, produced with very low Pt metal deposition levels (9APt/C), were found to have circumferentially wound single DNA double helix size surface fibers in the range of 30A width. Measurements of torus inner and outer circumference as well as ring thickness were performed. Observed differences in the torus dimension distributions from circular phi X-174 DNA and linear phi X-174 DNA may be related to the different topological constraints on DNA folding in these two samples (1). On the basis of annulus thickness measurements phi X-174 DNA toruses, in contrast to lambda DNA toruses, were observed to fall into two classes identified as being formed from monomer DNA condensation and multimer DNA condensation. All of the torus substructure and population dimensions observed here are consistent with the continuous circumferential DNA winding model of torus organization proposed by Marx and Reynolds (1) to explain the micrococcal nuclease cleavage properties of the toruses. End-on view measurements of the torus thickness were made from micrographs obtained by extensive tilting of the object replica. These direct measurements confirmed quaternary structure interpretations made from simple strand packing models. We compared the measured torus properties in this linear DNA size series (5386-48000 bp). With increasing DNA length the pattern of DNA strand self-assembly was found to be more varied producing lambda DNA toruses of varying shape. The relevance of our study to the problem of lambda bacteriophage DNA head packaging was discussed. PMID- 2978735 TI - Quaternary structure and function in phage lambda repressor: 1H-NMR studies of genetically altered proteins. AB - The quaternary structure and dynamics of phage lambda repressor are investigated in solution by 1H-NMR methods. lambda repressor contains two domains separable by proteolysis: an N-terminal domain that mediates sequence-specific DNA-A binding, and a C-terminal domain that contains strong dimer and higher-order contacts. The active species in operator recognition is a dimer. Although the crystal structure of an N-terminal fragment has been determined, the intact protein has not been crystallized, and there is little evidence concerning its structure. 1H-NMR data indicate that the N-terminal domain is only loosely tethered to the C-terminal domain, and that its tertiary structure is unperturbed by proteolysis of the "linker" polypeptide. It is further shown that in the intact repressor structure a quaternary interaction occurs between N-terminal domains. This domain-domain interaction is similar to the dimer contact observed in the crystal structure of the N-terminal fragment and involves the hydrophobic packing of symmetry-related helices (helix 5). In the intact structure this interaction is disrupted by the single amino-acid substitution, Ile84----Ser, which reduces operator affinity at least 100-fold. We conclude that quaternary interactions between N-terminal domains function to appropriately orient the DNA-binding surface with respect to successive major grooves of B-DNA. PMID- 2978736 TI - Glomerulonephritis and chronic myelogenous leukemia. AB - An adolescent with chronic myelogenous leukemia (CML) developed gross hematuria. Evaluation included renal biopsy revealing a proliferative glomerulonephritis with mesangial deposits of immunoglobulins A and G. An association between CML and immune-complex glomerulonephritis has not been previously reported and may represent a paraneoplastic phenomenon. PMID- 2978737 TI - Rapid haemodynamic responses to atrial natriuretic factor (99-126) in conscious sheep. AB - 1. Haemodynamic effects of 20 micrograms and 100 micrograms injection of atrial natriuretic factor 99-126 (ANF) were studied in conscious sheep. 2. ANF injection rapidly decreased blood pressure associated with a fall in total peripheral resistance, increased heart rate and cardiac output. These parameters returned to normal within 5 min of injection. 3. This study shows that ANF has an initial vasodilatory action to decrease blood pressure, which is different from the hypotensive mechanism seen with short-term infusion (60 min) of ANF in sheep. PMID- 2978738 TI - Atrial natriuretic factor in human pathophysiology. AB - 1. Evidence from numerous experiments incorporating central blood volume expansion and changes in sodium status supports atrial stretch as the prime determinant of ANF release. 2. Plasma ANF levels are the result of both secretion and clearance of the peptide. Clearance is altered by a number of factors, including changes in posture in normal man and is probably impaired in disease states with diminished renal and hepatic blood flow. 3. In normal subjects an inverse relationship exists between plasma ANF values and renin-angiotensin aldosterone system activity. This relationship is lost and replaced by a positive association in heart failure, presumably reflecting the abnormal concurrence of increased atrial stretch and diminished renal perfusion in this condition. Plasma ANF values rise with increasing severity of heart failure and fall with effective treatment. 4. Plasma ANF values are elevated in hypertension and cardiac tachyarrhythmias possibly reflecting raised central venous and atrial pressures. 5. A variety of other disorders may be associated with abnormal plasma ANF values including cirrhosis and the syndrome of inappropriate ADH secretion. 6. Evidence from low-dose infusions of ANF in normal volunteers suggests that the variations in plasma ANF seen in health and disease are sufficient to exert biological effects. 7. The advent of a specific antagonist is needed to provide further insight into the physiological and pathophysiological roles of ANF. PMID- 2978739 TI - Effect of antihypertensive therapy on the cardiovascular amplifiers. AB - 1. In established chronic hypertension the amplifier properties of vessels and heart contribute about 70% to the maintenance of the elevated blood pressure (BP). Recent studies in spontaneously hypertensive rats (SHR) suggest that the structural changes occur very early and their amplifier properties may be critical for the development of hypertension. 2. In patients with primary hypertension, the greater the regression of cardiac and vascular hypertrophy, the slower the subsequent redevelopment of hypertension. Following regression of hypertrophy, the antihypertensive action of moderate regular exercise can maintain BP in the normal range in a proportion of patients. 3. Early treatment of SHR with enalapril greatly reduces the subsequent 'steady-state' BP in SHR. This produces virtually complete regression of vascular hypertrophy, but somewhat lesser degrees of regression of cardiac hypertrophy. 4. These studies serve as models for primary and secondary prevention of hypertension. A strategy based on intermittent drug and non-pharmacological therapy in man may contribute to the secondary prevention of atherosclerosis, in view of the adverse effects on lipid profiles of many antihypertensive drugs. PMID- 2978742 TI - Response of atrial natriuretic peptide to adrenaline and noradrenaline infusion in man. AB - 1. Atrial natriuretic peptide (ANP) levels were significantly increased during both adrenaline and noradrenaline infusions, in the physiological range, in normal subjects and in patients with essential hypertension. 2. During adrenaline infusion significant increases in both circulating adrenaline and noradrenaline levels were observed. Mean arterial pressure was unaltered. Changes in heart rate were not significant. 3. During noradrenaline infusion, significant increases in circulating noradrenaline and mean arterial pressure were also observed. Heart rate and plasma adrenaline levels were unaltered. 4. Fluctuations in sympathetic nervous system activity may be involved in the regulation of ANP via adrenoceptor stimulated release of ANP. Other known regulators such as atrial stretch and increasing heart rate may modify this response. PMID- 2978740 TI - Red blood cell ionized calcium concentration in spontaneous hypertension: modulation in vivo by the calcium antagonist PN 200.110. AB - 1. Altered calcium regulation has been observed in experimental and human hypertension. In this study erythrocyte (RBC) intracellular calcium concentration ([Ca2+]i) was compared in conscious spontaneously hypertensive rats (SHR) and their normotensive controls (WKY) at rest and after injection of the dihydropyridine calcium antagonist PN 200.110. 2. Resting [Ca2+]i was similar in SHR and WKY. 3. PN 200.110 administration induced a rapid decrease in blood pressure in SHR and WKY. Five minutes after the injection no change in [Ca2+]i was observed; at 1 h [Ca2+]i was significantly decreased in SHR, but not in WKY. 4. These results suggest that the mutual adaptation of the rate of calcium influx through calcium channels and the activity of the calcium extruding pump differ between WKY and SHR. PMID- 2978741 TI - Decreased renal responses to atrial natriuretic peptide in renal wrap hypertension. AB - 1. The effects of atrial natriuretic polypeptide (ANP) on renal function were examined in renal wrap hypertensive rabbits and sham operated rabbits. 2. ANP (2 micrograms/min) induced hypotension, but did not produce significant diuresis, natriuresis or change in the glomerular filtration rate (GFR) in renal wrap hypertensive rabbits (n = 8). 3. In sham operated normotensive rabbits (n = 4), ANP induced significant diuresis (230%) and increased GFR by about 40%. 4. Thus, ANP was markedly less effective in the impaired kidneys of renal wrapped rabbits than in normal kidneys. PMID- 2978743 TI - Effects of volume expansion and contraction on plasma levels of atrial natriuretic peptide in man. AB - 1. Effects of saline infusion and blood removal on atrial natriuretic peptide (ANP) in normal subjects were examined in order to better define the magnitude of acute central volume regulatory influences on ANP. 2. Plasma ANP levels increased progressively during volume expansion with saline infusion, increasing by 18% after 30 min and by 93% after 120 min, and did not change during recumbency alone. 3. Plasma ANP levels immediately after a standard blood donation performed semirecumbent were significantly lower than before blood donation; they fell by 18%. 4. The magnitude of the fall in ANP induced by blood donation correlated significantly with basal plasma ANP. 5. In man, ANP responds to both increases and decreases in central blood volume, consistent with a role for ANP in blood volume homeostasis. PMID- 2978744 TI - Attenuated renal response to atrial natriuretic peptide infusion in rats with heart failure. AB - 1. The natriuretic and diuretic effects of three atrial natriuretic peptide (ANP) infusion rates were examined in rats 4 weeks after myocardial infarction induced by left coronary artery ligation. 2. The natriuretic and diuretic effects of ANP were observed in controls and rats with infarction, but the effects were significantly attenuated in the latter. 3. Rats with chronic left heart failure were less sensitive to the renal effects of ANP compared with controls. 4. Impaired sodium and water excretion in chronic heart failure may be due partly to an attenuated renal response to ANP. PMID- 2978745 TI - Altering angiotensin levels by administration of captopril or indomethacin, or by angiotensin infusion, contributes to an understanding of atrial natriuretic peptide regulation in man. AB - 1. Plasma atrial natriuretic peptide (ANP) levels were positively correlated with plasma renin activity (PRA) levels, when blood volume and blood pressure (BP) were not raised in normal subjects (NLS) or patients with postoperative aldosterone-producing adenoma (APA), Bartter's syndrome (BS), Addison's disease, anorexia nervosa, diuretic abuse or salt-losing congenital adrenal hyperplasia. 2. Angiotensin II infusion raised ANP levels in NLS, and patients with BS, pre- and postoperative APA, only when BP rose, suggesting that this effect might be mediated by the rise in BP. 3. Captopril lowered aldosterone and ANP levels in renal artery stenosis, but falling BP levels could mediate this effect. Captopril lowered aldosterone and BP in BS, but did not lower ANP, perhaps because angiotensin remained elevated. 4. Indomethacin lowered ANP when PRA was initially normal or raised (NLS and BS), but not when PRA was suppressed (APA). This effect could not be mediated by BP, which rose, but could be mediated by renin angiotensin, which fell. 5. Factors other than central blood volume and atrial stretch may modulate ANP levels. Plasma angiotensin II may be such a factor, and may exert an important influence at high levels, especially when blood volume is low. PMID- 2978746 TI - Effect of glycaemic control on glomerular filtration rate in the streptozotocin diabetic rat. AB - 1. Diabetes was induced in 32 adult Wistar-Kyoto rats with streptozotocin (60 mg/kg). Fourteen rats remained untreated, 10 received insulin three times per week, and eight received insulin daily. Fourteen non-diabetic rats served as controls. 2. Exchangeable sodium and plasma volume were elevated in the untreated diabetic rats. Treatment normalized these parameters. 3. Glomerular filtration rate (GFR) was elevated in the untreated diabetic group and the group receiving insulin three times per week compared with the control group. Daily insulin treatment restored GFR towards control values. 4. Plasma atrial natriuretic factor was similar in untreated diabetic rats and non-diabetic rats. PMID- 2978747 TI - Renal effects of atrial natriuretic factor (99-126) in conscious sodium-replete sheep. AB - 1. The effect of renal arterial infusion of synthetic human atrial natriuretic factor (ANF (99-126] on renal function in the conscious euvolaemic sheep was characterized. ANF (99-126) was infused for 2 h at 5 and 50 micrograms/h into the renal artery of crossbred Merino ewes with chronically indwelling cannulae inserted in the renal artery. The effect on absolute and fractional excretion of Na, K, Ca, Cl and HCO3, glomerular filtration rate (GFR), effective renal plasma flow (ERPF) and free water clearance (CH2O) were measured. 2. Infusion at 50 micrograms/h produced a fourfold increase in Na and Cl excretion. Ca excretion increased eightfold, while K and HCO3 increased by small amounts. At the lower dose only Na, Cl and Ca excretion increased significantly. The changes in absolute excretion of each ion were closely mirrored by changes in fractional excretion. CH2O became more negative at both levels of infusion. Small changes in GFR were measured at both rates of infusion. No changes in ERPF or renin secretion were observed. 3. ANF (99-126) infusion at 50 micrograms/h for 1 h increased the excretion of Li, such that more than 70% of the change in Na excretion was associated with the changes in Li clearance. Changes in GFR accounted for less than 10% of change in Na excretion. 4. Following either long term (50 micrograms/h for 6 h) or repeated short-term (20 micrograms/h for 30 min) infusions of ANF (99-126), the response of Na excretion was not sustained. The mechanisms of the tachyphylaxis remains undetermined. 5. ANF (99-126) is a powerful stimulus to the absolute and fractional excretion of Na, K, Ca, Cl and HCO3. The mechanism of action is not known, but appears to be related to changes in tubular function and/or a change in glomerulotubular balance. PMID- 2978748 TI - Interaction between atrial natriuretic factor and ouabain: vascular reactivity to noradrenaline in pentobarbital anaesthetized dogs. AB - 1. The influence of intra-arterial infusion of rat atrial natriuretic factor (ANF 8-33) and/or ouabain on the vascular responses to noradrenaline was investigated in the denervated and flow-controlled hindlimb preparations in pentobarbital anaesthetized dogs. 2. During the continuous infusions of ANF (30-40 min) vascular responses to noradrenaline were significantly depressed. Subsequent infusion of ouabain together with ANF (50-60 min) reversed and restored the vascular reactivity to the control levels. Hypotension produced by ANF infusion was partially reversed during the simultaneous infusions of both the agents. 3. In a separate series of experiments, in which ouabain was first infused (50-60 min) vascular responses to noradrenaline were significantly enhanced. Subsequent infusions of ANF (plus ouabain) even up to 60 min or longer failed to alter the enhanced vascular responsiveness facilitated by ouabain. 4. The present studies demonstrate a physiological antagonism between ANF and ouabain and such a phenomenon could account for the previous observation that vascular reactivity to noradrenaline was progressively enhanced after acute blood volume expansion. Whereas plasma levels of both ANF and ouabain-like inhibitor(s) of the sodium pump are elevated after volume expansion, inhibitory effects of ANF on the vascular smooth muscle may be compromised in the presence of an Na+ pump inhibitor(s). PMID- 2978750 TI - Community service. Part III of a series. PMID- 2978749 TI - Evaluation of a pediatric dentistry curriculum by graduates. PMID- 2978751 TI - Pediatric dentistry comes of age. PMID- 2978752 TI - Shear bond strengths to dentin: effects of surface treatments, depth and position. PMID- 2978753 TI - [Periodontal condition of 15 children with acute lymphatic leukemia or Hodgkin's lymphoma before and after induction therapy]. PMID- 2978754 TI - Qualitative and quantitative strategies for exploring the progress of sex education for the handicapped. PMID- 2978756 TI - A system for delivery of dental care in the long-term care facility. PMID- 2978755 TI - Wellness: a valuable resource for persons with disabilities. PMID- 2978757 TI - Referring medically compromised and infectious dental patients. PMID- 2978759 TI - [The dance of AIDS]. PMID- 2978758 TI - Immunological abnormalities of acquired immunodeficiency syndrome and related disorders in patients from Rio de Janeiro, Brazil. AB - The immunological profile of acquired immunodeficiency syndrome (AIDS) and chronic lymphadenopathy syndrome (CLAS) in 15 and 11 Brazilian patients, respectively, was studied. The AIDS patients showed reduced percentage of total T (CD3) and T-helper-inducer (CD4) lymphocytes, relative increase in numbers of T suppressor-cytotoxic (CD8) cells and a marked inversion of T-helper inducer/suppressor-cytotoxic (CD4/CD8) ratio. Lymphoproliferative responses to PHA, ConA, PPD and PWM were diminished. Hypergammaglobulinemia and high levels of circulating immune complexes were also found. The CLAS patients also showed important immunological alterations, but not so intense as those with AIDS. These data seems to be similar to those observed in other parts of the world. PMID- 2978760 TI - [Effect of several resin monomers on glucosyltransferase activity of Streptococcus mutans]. PMID- 2978761 TI - Behavioral influences of rectal diazepam in solution on dental patients with mentally and physically handicapping conditions. AB - The hypothesis of this study was to determine whether the use of rectal diazepam in solution would effectively modify the uncooperative behavior of patients with mentally and physically handicapping conditions during dental treatment. The sample consisted of 42 patients with mild to severe mentally handicapping conditions, 4 to 31 years old, who live in a homecare center. Supragingival and subgingival scaling and prophylaxis were attempted during a 5-minute period and the patient's behavior was assessed. Conscious sedation using a rectal solution of diazepam (Stesolid) was used for those patients with voluntary or involuntary uncooperative behavior that prevented treatment. Twenty-two subjects (52.4%) were treated without diazepam. The rectal solution of diazepam proved to be a significantly (P less than .01) effective agent for behavior modification permitting the successful treatment of 16 (80%) of the 20 remaining subjects. The following behaviors were significantly modified: places hand(s) or arm(s) in front of mouth (P less than .01); does not open mouth, lips held firmly together (P less than .05); turns head to one side or side to side (P less than .05); attempts to grab instruments (P less than .05). No relationship was found between the results and the variables of medical history, gender, age, weight, quantity of diazepam administered, routine medication, pulse rate, blood pressure, and respiration. PMID- 2978762 TI - Oral manifestations of Beckwith-Wiedemann syndrome. AB - A report of case of a patient with Beckwith-Wiedemann syndrome is presented. The macroglossia associated with the syndrome necessitated long-term dental follow-up to prevent and treat problems with occlusion, speech, and esthetic appearance that might develop. Early intervention might be required if feeding or respiratory difficulties, resulting from the macroglossia, arise. The diagnosis of the syndrome should alert the dentist to the increased incidence of intra abdominal malignancies and the need for periodic observation. PMID- 2978763 TI - Development of a hospital-based dental phobia clinic. PMID- 2978764 TI - Use of the periodontal ligament injection in dental care of the patient with hemophilia--a clinical evaluation. AB - The PDL injection can be effective when used in the dental patient who has hemophilia. In the case reported here, no postoperative complications related to hemorrhage or hematoma formation were recorded. The PDL injection offers the patient with hemophilia improved anesthetic reliability without factor administration for routine dental procedures. PMID- 2978765 TI - Dental considerations for the patient receiving dialysis for renal failure. AB - A review of the literature describing the dental management of patients receiving hemodialysis because of renal failure is presented. A description of the renal failure process is given. Pretreatment and day of treatment precautions are considered. A pertinent dental case report of a dialysis patient is also presented. PMID- 2978766 TI - Prevention of self-inflicted trauma: dental intervention to prevent chronic lip chewing by a patient with a diagnosis of progressive bulbar palsy. AB - An appliance is used as a beginning in the prevention of self-mutilative oral behavior. The patient in the case reported here required a quick solution to a severe problem of self-inflicted trauma. The design presented here can be improved on or an altogether new approach may be developed. PMID- 2978767 TI - Behavioral and pharmacological dental management of a patient with autism. AB - A 10-year longitudinal report of case of an institutionalized autistic male dental patient is described. Interpretation of the data in the clinical and medical charts during this period points to possible clues to facilitating treatment for autistic individuals. One potentially important finding is the apparent inverse relationship that was found between level of sedation and patient cooperation. The impact of the specific medical diagnosis on the comprehensive care for these patients is also addressed. PMID- 2978768 TI - Guidelines for the use of local anesthetics in the dental treatment of patients who are susceptible to malignant hyperthermia. AB - There is controversy regarding the safe use of local anesthetics in patients who are susceptible to malignant hyperthermia undergoing dental treatment. This article reviews the literature for reports of malignant hyperthermia reactions under local and general anesthesia, and suggests a protocol for the management of these patients in the dental office. PMID- 2978769 TI - The central indwelling venous catheter in the pediatric patient--dental treatment considerations. AB - Dentists need to understand and recognize the special needs of this unique patient population before dental therapy is instituted. The primary objective of this paper is to familiarize the dental practitioner with the central venous catheter and its indications in pediatric medicine. In addition, recommendations concerning the appropriate management of these patients are provided. PMID- 2978770 TI - A comparison of two mobile treatment programs for the homebound and nursing home patient. AB - Two mobile treatment programs using portable equipment transported in vans to serve homebound persons in Denver and Chicago are compared for types of patients treated, use by local dentists, types of services provided, fees generated, and costs involved in operation during the 2-year period (1985-86). Both programs treated a similar, largely nursing home-based white female population that was predominantly older. Volunteer dentist participation varied greatly, with more than twice the number of dentists using the service in Chicago. Both programs accomplished essentially the same number of visits for the biennium studied, with 1,324 for Chicago and 1,320 for Denver. The Denver program was more efficient, generating more visits each time a dentist used the program. The services provided in total were about the same for both programs, with Denver generating 4,887 procedures and Chicago 4,602 for the biennium, but Denver had a more favorable ratio of diagnostic to treatment services. The costs of both programs were close, averaging about $60,000 per year. Denver was able to generate far more in equivalent fees than Chicago for the biennium, but Chicago dentists donated a greater percentage of services (67) than did Denver dentists (62). PMID- 2978771 TI - Maternal perceptions of child behavior: handicapped versus nonhandicapped. AB - This study compares the ratings of child behavior in short stories by mothers of children with handicapping conditions versus those whose children had no handicapping conditions. Each of the mothers rated 12 short stories or vignettes and indicated their self-perceived level of stress. The results showed that the mothers of children with handicapping conditions expressed significantly greater stress levels and rated inappropriate child behavior in a more tolerant fashion than the mothers of children without handicapping conditions. This information may be of value to the dentist who treats patients with handicapping conditions, and it provides a greater understanding of the mothers' concerns and daily life stresses. PMID- 2978772 TI - The patient with cancer: social and clinical perspectives for the dentist. AB - Dentists can have a significant role in the care of individuals with cancer. They detect and provide treatment for patients with head and neck cancer and routinely provide oral care to patients who have other forms of cancer. This paper addresses the social, clinical, and psychological responses to cancer and considers the patient's understandings of prevention, diagnosis, and treatment. The findings in this paper are based on intensive interviews with 42 patients with cancer and family members, sociological participant-observation of oral and maxillofacial rehabilitation of the patient with head and neck cancer in major teaching hospital settings, and a review of the literature. Cancer was associated with an uncertain course and prognosis, made more problematic by imagery, fear, and fatalism. Patient behaviors, including delayed care seeking, are related to understandings of cancer and the social and family interactions that may occur around the disease. Professional role dilemmas, strategies toward informing patients of their cancer, and dealing with death are considered as issues dentists may face in care of the patient with cancer. PMID- 2978773 TI - Fevers of unknown origin in patients requiring mechanical ventilation: an odontogenic problem? AB - Patients who are on continuous monitors and mechanical ventilators, and who are receiving medical care in an intensive care setting, do not meet the usual expectations for dental care. Two examples of odontogenic contribution to fevers of unknown origin are presented in the context of treatment of continuously monitored patients with mechanically supported respiration. It is intended that these cases show the need for active dental consultation and participation by dentists in the care of the patient who is chronically ill and who cannot be brought to the traditional dental care setting. PMID- 2978774 TI - Oral complications of cancer chemotherapy: prevention and management. AB - Cancer chemotherapy is used to destroy rapidly proliferating cells. However, normal cells with high mitotic indexes are also affected by chemotherapy, particularly those in the oral and gastrointestinal mucosa and the hemopoietic system. Ultimately, this may lead to certain oral complications of cancer chemotherapy such as mucositis, infection, hemorrhage, xerostomia, and neurologic and nutritional disorders. The prevention and management of these oral complications that may result from the chemotherapeutic management of cancer are discussed here. PMID- 2978775 TI - Dental treatment needs among hospitalized adult mental patients. AB - This survey determines the dental care needs of hospitalized adults with psychiatric and mental disorders. Comprehensive clinical and radiographic oral examinations, conducted on 33% of the patients in a large state mental hospital, found that extensive unmet needs existed in this population. The major requirements were for prophylaxis, calculus removal, and periodontal therapy. The patients' needs varied depending on several demographic factors, including length of hospitalization and psychiatric diagnoses. PMID- 2978776 TI - Feeding disorders research and teams: a new challenge for dentistry. AB - Feeding disorders, which include masticatory and cerebral disabilities, present a major problem for disabled persons, but only in the last decade have health professionals shown an increased interest in the diagnosis and management of patients with such problems. Dentists are experts in the diagnosis and treatment of disorders of the oral-facial region, yet they have either worked in intradisciplinary isolation or have not demonstrated their interest in patients with feeding disorders. Consequently, those involved in the assessment and treatment of persons with feeding disorders are not aware of the contributions dentists have made to the basic and clinical data base of feeding assessment and therapy. Dentistry is simply not mentioned in rehabilitation. The opportunity now exists for dentistry to inform others of our expertise in the field from both a research and a clinical perspective. The basic and clinical base of dental therapeutics and its role in the diagnosis and treatment of disorders of the masticatory apparatus and of swallowing are reviewed here. It is hoped that this review will stimulate interest among dentists to take part in the rehabilitation of patients with feeding disorders through the use of the extensive data base within the profession. PMID- 2978777 TI - Interim dentures for the older patient who is medically compromised. AB - The concept of interim or transitional dentures is appropriate to the treatment of older patients who are hospitalized or medically compromised. This paper briefly reviews the emotional and physiological needs of these patients, and illustrates a case in which an interim denture technique was beneficial. PMID- 2978779 TI - Ensuring the rights of persons with disabilities. AB - The following keynote address by Justin Dart, chair, congressional Task Force on the Rights and Empowerment of Americans with Disabilities, was presented Aug 8 in Philadelphia during the ninth congress of the International Association of Dentistry for the Handicapped. PMID- 2978778 TI - Agoraphobia: review and considerations in dental treatment. AB - The nature of agoraphobia as well as problems that may be encountered during dental treatment of the patient with agoraphobia is discussed to aid dentists' recognition and understanding of the syndrome. PMID- 2978780 TI - Initial results of the use of a drug-interaction system. AB - Dentists' knowledge about drug interactions, particularly in medically compromised older patients, may not be adequate. An on-line drug interaction system was tested for 4 months in a dental clinic to ascertain its usefulness in training students. Characteristics of high-risk patients are described, as well as opinions of users about the applicability of information presented and its relation to patient care. Approximately 3% of the patients seen were at risk of trauma from drug interactions, half of which were considered likely to occur. Implications of drug interaction data are also discussed in relation to treatment planning. PMID- 2978781 TI - Wheelchair transfer techniques for the dental office. AB - Dental offices must be made more accessible to individuals who are physically challenged. Transfer techniques for safely moving patients between wheelchairs and dental chairs, sliding transfers, one- and two-person transfers, and methods of accommodating patients in a dental operatory who cannot be transferred from wheelchairs are presented. PMID- 2978782 TI - Oral hygiene for the severely handicapped: clinical evaluation of the University of Mississippi dental care system. AB - The study reported here was a clinical evaluation of an automatic toothbrushing device for persons with severe handicapping conditions. This device, the University of Mississippi dental care system, is designed for patients who are unable to control their arms and hands. The effectiveness of this device was compared to routine (ordinary, manual) regimens of oral hygiene through use of gingival (Loe-Silness) and plaque (Turesky) indexes (GIs, PIs). Plaque indexes for patients under routine care averaged 2.11; PIs for patients under the dental care system averaged 1.47. Based on Ebbutt's extra period changeover design model, the t-test for difference in plaque scores is significant (P less than .001). The results indicate that the University of Mississippi dental care system improves the patients' ability to independently control plaque. PMID- 2978783 TI - Special oral concerns in people with severe and profound mental retardation. AB - Dental patients with severe and profound mental impairments may engage in behaviors that adversely affect their oral health. Based on experience in a residential care facility, this report sensitizes dental practitioners, who are unfamiliar with the problems of these patients, to consequences of these behaviors. PMID- 2978784 TI - Spironolactone therapy in hirsutism and acne. PMID- 2978785 TI - Surgery for rhinophyma: a case report. PMID- 2978786 TI - Identification of a neuronal laminin receptor: an Mr 200K/120K integrin heterodimer that binds laminin in a divalent cation-dependent manner. AB - Neuronal interactions with extracellular matrix (ECM) components are crucial for axon growth and guidance during development and nerve regeneration. Laminin (LN), a prominent ECM glycoprotein, promotes neuronal survival and axon growth. To identify neuronal receptors for LN, we looked for cell surface proteins on the neuronal cell line B50 that bind LN. An integrin alpha/beta 1 dimeric receptor was identified and purified using lectin and LN affinity chromatography. The purified integrin contains two subunits with Mrs of 200 K and 120 K that bind LN specifically in the presence, but not the absence, of divalent cations (Ca2+/Mg2+ or Ca2+/Mn2+). The Mr 120 K protein was identified as the rat integrin beta 1 subunit using two beta 1 subunit-specific antibodies, and was shown to form a noncovalent complex with the Mr 200K putative alpha subunit. Since neurons and neuronal cell lines express similar integrin beta 1-class heterodimers that mediate attachment and process outgrowth on LN, the Mr 200K/120K complex identified here is likely to be an important laminin receptor used by neurons. This integrin may also mediate binding to LN by many nonneuronal cell types. PMID- 2978787 TI - Cis-trans interconversion of acitretin in man. AB - The major plasma metabolite of acitretin (trans-acitretin) is its 13-cis isomer, cis-acitretin. Interconversion of cis-acitretin to trans-acitretin was demonstrated in man following administration of a single oral dose of cis acitretin. Plasma concentrations of Ro 13-7652 (cis-acitretin) and Ro 10-1670 (trans-acitretin) were much higher after cis-acitretin administration than after trans-acitretin administration. Surprisingly, these high concentrations were not associated with a clear therapeutic effect in dermatoses (e.g. psoriasis) which are usually responsive to oral retinoids. Interactions between the cis and trans isomers formed in vivo may explain the difference in therapeutic activity of each stereoisomer when administered orally. PMID- 2978788 TI - Giant left atrium and mitral valve replacement: risk factor analysis. AB - Giant left atrium (GLA) associated with mitral valve disease (MVD) has been reported as a significant risk factor in mitral valve surgery with mortality ranging from 8%-32%. Plication of the left atrium has been suggested to reduce postoperative left ventricular failure, respiratory failure and mortality. The 203 consecutive patients with MVD operated upon between 1980 and 1986 were reviewed and divided in two groups: group A without GLA (165 patients) and group B with GLA (38 patients = 19%). The pertinent preoperative and intraoperative notes and the early and late postoperative course were reviewed and correlated. The hospital mortality was 2.4% in group A and 2.6% in group B. Late mortality, at a mean follow-up of 54 months was 4.3% in group A and 5.4% in group B. In this series, GLA was not a significant risk factor in MVR and did not affect early and late results as compared with cases without GLA. Plication may not be required in absence of extracardiac signs of compression. PMID- 2978789 TI - Assessment of hemodynamic function and tolerance to ischemia in the absence or presence of calcium antagonists in hearts of isoproterenol-treated, exercise trained, and sedentary rats. AB - The effects of cardiac hypertrophy on the structure, function and tolerance to ischemia of rat hearts have been investigated. Multiple injections of low doses of isoproterenol (ISO) resulted in an increase of heart weight/body weight ratio by 60%, and a decrease of myocardial creatine kinase activity by 25%, as compared to normal rats. Compared to age-matched control rats, rats submitted to a swimming program had a higher heart weight by 20%, but similar values of heart weight to body weight ratio. In isolated perfusion, the functional capacities of hearts from ISO-treated rats were severely depressed compared to normal rat hearts whereas exercise-trained rat hearts performed as well or even better than control hearts. The functional recovery of ISO-treated hearts following cardioplegia-induced arrest for 20 min at 37 degrees C was significantly worse than the recovery of normal hearts, but hearts of exercise-trained rats showed a significantly better recovery than control hearts. Exercise training results in improvement of myocardial blood supply resulting in better preservation of the heart during ischemia, compared to normal hearts. Addition of a combination of verapamil and diltiazem to the cardioplegic solution followed by ischemic arrest for 20 min at 37 degrees C resulted for ISO-treated rat hearts in an improved recovery of cardiac output (99%) compared to cardioplegia in the absence of these drugs (72%). In exercise-trained and control rat hearts, calcium antagonists improved the recovery from cardioplegic arrest of cardiac output from 90% to 92% and from 71% to 87%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2978790 TI - Axillo-femoral bypass failure secondary to axillary stenosis: ultrasound-guided percutaneous transluminal angioplasty. PMID- 2978791 TI - Progesterone decreases DNA binding factor activity and the expression in Xenopus oocytes of a cAMP responsive gene from rat liver. AB - Stage VI Xenopus oocytes were injected with a plasmid (pBB0.6-CAT) which contains the cAMP regulatory element (CRE) from the rat liver phosphoenolpyruvate carboxykinase (PEPCK) gene fused upstream from a reporter gene [chloramphenicol acetyltransferase (CAT)]. Inhibition of the expression of the reporter gene (average = 51%) was observed in the presence of 10 microM progesterone, which is known to lead to inactivation of the oocyte cAMP dependent protein kinase (A kinase). In contrast, oocytes injected with a control plasmid (pSV2CAT), which contains no CRE, exhibited a variable increase (average = 31%) in CAT activity after progesterone treatment. Injection of the purified bovine cardiac A kinase catalytic subunit prior to exposure of oocytes injected with pBB0.6 CAT to progesterone prevents the loss of CAT activity generated by incubation with the steroid. Gel retardation analyses with oocyte lysates and a labeled synthetic oligonucleotide fragment containing the CRE from the PEPCK gene showed the existence of a complex with the same Rf and specificity as that formed with rat liver extracts. Subsequent exposure to progesterone, however, led to a rapid and extensive decrease in this binding activity. Taken together, these results are consistent with but do not prove the hypothesis that progesterone treatment and A kinase inactivation lead to a decrease in pBB0.6 CAT expression by virtue of a decline in the binding activity of an oocyte factor(s) to the CRE of the PEPCK fragment in pBB0.6-CAT, thereby decreasing transcription of the CAT gene. PMID- 2978793 TI - [Regional supply of orthodontists and pedodontists by specialty]. PMID- 2978792 TI - Plasma membrane and cytoskeletal constituents in Leishmania mexicana. AB - The presence and the localization of actin, spectrin and ankyrin are studied by immunofluorescence and immunoblotting in Leishmania mexicana promastigotes growing in vitro. These proteins, amphitropic in nature, coexist both in soluble and insoluble forms. Our results demonstrate that the Triton insoluble form of these proteins constitutes beside tubulin the cytoskeletal scaffold of promastigotes in close association with the plasma membrane, the axoneme and the basal body of the parasite. PMID- 2978794 TI - [Management of the hemophilic child]. PMID- 2978795 TI - [Assignments of dental team members in treatment of patients at risk]. PMID- 2978796 TI - [Detection and prevention of oral complications in diabetics]. AB - Oral affections are important causes of disorders in diabetes, within them, periodontal disease is the most common one, which comprises gingivitis and periodontitis. Dental plaque, high concentrations of salivary calcium and glucose, hyperglycemia and a lower resistance to infections, are main factor contributing to periodontal disease, dental caries, mycotic stomatitis and aphthae. Even though with the most advanced surgical methods and using antibiotics, systematically, the treatment of periodontal disease is rather difficult and often unsuccessful, therefore, to prevent it is the best treatment available, hence the importance of teaching oral health to the diabetic patient, which should comprise a good metabolic control, right tooth-brushing and visits to the stomatologist every six months or in the presence of gingival bleeding or gingival pus emanation. Diabetic condition is not a contraindication for the extraction of carious dental teeth, on the contrary, such teeth must be extracted when required or adequately treated. PMID- 2978797 TI - [A survey on dental diseases among handicapped children in pedodontic outpatient clinic and special clinic for handicapped children. 2nd report: IV, V, VI, and VII groups according to Uehara's classification]. PMID- 2978798 TI - [Toothbrushing of physically handicapped children (2)]. PMID- 2978799 TI - [Studies of the facial features of Down's syndrome by Moire topography]. PMID- 2978801 TI - [Clinical application of ampicillin suppository (KS-Rl) in pedodontic clinic]. PMID- 2978800 TI - [Investigation concerning the children with congenital heart disease]. PMID- 2978802 TI - [The effectiveness of intravenous flunitrazepam administration with inhalation sedation using N2O-O2 in dental treatments of handicapped patients]. PMID- 2978803 TI - Spondylolisthesis. A common cause of back pain in Greenland. PMID- 2978804 TI - Lipoprotein (a) as a risk factor for atherosclerotic diseases. PMID- 2978805 TI - Health associated risk factors for academic achievement in a group of Manitoba Indian children. PMID- 2978806 TI - Initiation of a hepatitis B vaccination program in the Northwest Territories of Canada. PMID- 2978807 TI - Echocardiographic and Doppler assessment of hypertrophic cardiomyopathy. AB - Echocardiographic and Doppler studies are invaluable in the detection and management of patients with hypertrophic cardiomyopathy, since virtually all aspects of the disorder can be evaluated. Echocardiography is the primary method of diagnosis, and hemodynamic studies are required usually only when surgery is contemplated. All patients should if possible undergo complete M-mode, two dimensional, Doppler, and color flow studies to define the extent of hypertrophy, the hemodynamic subgroup, quantitation of left ventricular outflow obstruction, mitral regurgitation when present, and an assessment of diastolic function. Patients undergoing medical or surgical therapy for this disorder should have repeat studies performed to assess the effects of therapy on the degree of obstruction and diastolic dysfunction. PMID- 2978808 TI - Comparison of peak and modal aortic blood flow velocities with invasive measures of left ventricular performance. AB - The purpose of this study was to determine the accuracy of Doppler-derived modal and maximum velocity and peak and mean acceleration of ascending aortic blood for the assessment of left ventricular systolic function. Studies were performed in six anesthetized open-chest dogs. Doppler-derived modal velocity, maximum velocity, and peak and mean acceleration were compared with left ventricular dP/dt, maximum aortic blood flow, and rate of blood flow measured with an electromagnetic flow probe under varying inotropic states. Maximum Doppler velocity showed better correlation (r = 0.94, y = 0.34 + 3.95) with maximum aortic blood flow than the modal velocity (r = 0.85, y = 1.49 + 3.85x). Peak acceleration also correlated better with the rate of blood flow (r = 0.92, y = 12.3 + 4.92x) than the mean acceleration (r = 0.83, y = 12.2 + 4.27x). Modal and maximum velocity and mean and peak acceleration correlated well with left ventricular dP/dt. We conclude that peak modal and peak maximum velocity and peak and mean acceleration are accurate measurements of left ventricular function. Maximum velocity and peak acceleration are more accurate than modal velocity and mean acceleration. PMID- 2978809 TI - Evaluation of reperfusion hyperemia with myocardial contrast echocardiography. AB - In this study we used myocardial contrast echocardiography to evaluate reperfusion hyperemia in an open-chest canine model of temporary coronary artery occlusion. Eight dogs had coronary occluders and electromagnetic flow probes on the left circumflex coronary artery. Aortic root injections of agitated sodium diatrizoate and saline solution were used for myocardial contrast. Data were collected at baseline (n = 16), during coronary occlusion (n = 18), immediately after coronary release (n = 18), and 5 minutes after coronary artery release (n = 12). Baseline coronary flow was 23.8 +/- 5.9 ml/min, decreasing to 0 ml/min during coronary occlusion. Immediately after coronary release flow was 96.6 +/- 41 ml/min (p less than 0.001 compared with baseline), and 5 minutes after coronary release flow was 68.2 +/- 27.9 ml/min (p less than 0.001 compared with baseline). The myocardial image intensity change after injection of contrast material was 74.25 +/- 30.6 ml/min at baseline and declined to 10.4 +/- 10.9 ml/min during coronary occlusion (p less than 0.001 compared with baseline). During reperfusion hyperemia image intensity change was 102.3 +/- 33.3 ml/min (p less than 0.001 compared with occlusion, p less than 0.02 compared with baseline, p less than 0.001 compared with remote regions). Considering all observations, myocardial image intensity change after contrast injection correlated positively with coronary flow (r = 0.67, p less than 0.001). Correlations within individual dogs ranged from r = 0.70 to 0.98. We conclude that image intensity change after aortic root injection of echocardiographic contrast correlates with coronary blood flow. Objective measurements of contrast intensity reflect increases in coronary flow associated with reactive hyperemia after coronary occlusion and release. PMID- 2978810 TI - Lipomatous hypertrophy of the interatrial septum: characterization by transesophageal and transthoracic echocardiography, magnetic resonance imaging, and computed tomography. AB - Lipomatous hypertrophy of the interatrial septum is a benign condition that must be distinguished from other space-occupying lesions of the atria. Patients with this disorder generally have chronic pulmonary disease and thus are difficult to image with conventional transthoracic two-dimensional echocardiography. Transesophageal echocardiography can provide high quality imaging of intracardiac structures in patients who lack adequate transthoracic echocardiographic windows as a result of pulmonary disease. This case report describes the appearance of lipomatous hypertrophy of the interatrial septum as investigated by transesophageal echocardiography. PMID- 2978812 TI - OK-432 and IL-2-augmented cytotoxicity of human natural killer cells and cytotoxic T lymphocytes at the clonal level. AB - The biological response modifiers OK-432 and interleukin-2 (IL-2) were found to enhance the lytic capacity of cloned CD3- natural killer (NK) cells and CD3+ T cells. With respect to NK cells, only those clones with a high proliferative capacity and cultured without phytohaemagglutinin (PHA) responded with enhanced lytic capacity to OK-432. OK-432, but not IL-2, was found to augment the antibody dependent cellular cytotoxicity of cloned NK cells. With T-cell clones, OK-432 augmented the cellular cytotoxicity of CD3+8+ but not that of CD3+4+ cytotoxic T lymphocytes, while IL-2 augmented the cytotoxicity of both types of clone. Taken together, these results indicate that OK-432-augmented lytic capacity is not restricted to NK cells and its pathway of action may be independent of IL-2. PMID- 2978811 TI - The role of C3 and its fragments in the control of S phase entry of activated mouse B lymphocytes via the complement receptor type 2. AB - Complement component C3 and receptors for its activated fragments have been recognized as regulators of humoral immune responses. Cross-linked C3b and C3d will substitute for alpha B cell growth factors (alpha BCGF) produced by macrophages by stimulating activated murine B lymphocytes to enter S phase. In contrast, soluble C3d and synthetic oligopeptides--containing the amino acid sequence of the binding site of C3d to its receptor, the complement receptor type 2--inhibit the entry into S phase of alpha BCGF-stimulated B cells. There is evidence supporting the hypothesis that a portion of the C3 gene at the 3' end, encoding parts of the alpha chain, encodes an alternate C3 product with alpha BCGF activity. PMID- 2978813 TI - Abnormal movements of the oral-facial region--diagnosis, assessment and control. A guide for the dental clinician. PMID- 2978814 TI - Paediatric dentistry. PMID- 2978815 TI - [Dental care for the handicapped]. PMID- 2978817 TI - Provision of full dentures following neurological surgery. PMID- 2978816 TI - [Rett syndrome: a "new" and rare disease. A clinical case]. PMID- 2978818 TI - AIDS, confidentiality and dental care. PMID- 2978819 TI - Enzymatic activities in gingival fluid with special emphasis on phospholipases. PMID- 2978820 TI - Progeria in a pediatric dental patient: literature review and case report. PMID- 2978821 TI - Mouthstick prosthesis placement in a 19-month-old arthrogryposis multiplex congenita patient: case report. PMID- 2978822 TI - Strategic planning in pediatric dentistry: the students' perspective. PMID- 2978823 TI - Conscious sedation practices in pediatric dentistry: a survey of members of the American Board of Pediatric Dentistry College of Diplomates. PMID- 2978824 TI - Abdominoplasty combined with total abdominal hysterectomy. AB - Twenty patients who underwent total abdominal hysterectomy combined with abdominoplasty are described here, along with the criteria used for their selection. No patients suffered pulmonary embolus, significant infection, or other serious complications. Combining the procedures shortened the convalescent period and reduced the total cost. In carefully selected cases it appears that these two procedures can be combined safely. PMID- 2978825 TI - Free microneurovascular gracilis muscle transfer in the dog for enterostomal sphincter construction and control: an experimental study. AB - Hundreds of thousands of individuals struggle today with the morbidity of an abdominal wall enterostoma. This study explored two ways of constructing abdominal wall sphincter mechanisms in dogs using gracilis muscles transferred by free microneurovascular technique. Transferred muscles successfully generated pressures greater than the normal external anal sphincter but could not maintain this level of contraction for tonic sphincter closure. Other transferred muscles were designed to straighten the course of stomas continent at rest. They provided strong linear pull uncoiling the stomas but simultaneously pinched off the orifices during contraction. This opening model shows promise if the muscle attachments around the stoma orifices could be made less constricting. PMID- 2978826 TI - Alternatives to the classic abdominoplasty. AB - Prior to designing and performing an abdominoplasty, we use a simple test that allows us to predict, within 1 cm, the location of the suprapubic scar after a standard abdominoplasty. When the distance from the umbilicus to the suprapubic scar is less than 10 cm, we recommend using different techniques, such as an abdominoplasty with a short vertical scar, a midabdominoplasty, or an abdominal up-and-down lift. We suggest that only a belt lipectomy can improve the silhouette appearance after a substantial weight loss. PMID- 2978827 TI - Selective colony blotting to expand bacterial surface receptors: applications to receptors for rat immunoglobulins. AB - Many bacterial surface receptors demonstrate a heterogeneous expression pattern among individual colonies. Methods have been developed to select bacteria expressing high levels of a stable surface receptor. This process is illustrated using a Streptococcus zooepidemicus isolate demonstrating a high level of Fc receptors for rat immunoglobulins. This strain was selected and expanded to obtain a bacterial isolate demonstrating approximately 100 fold greater reactivity with rat immunoglobulins than protein A positive Staphylococcus aureus or 30-40 fold higher reactivity for rat IgG than type III Fc receptor positive streptococcal group G strains. The optimal pH for rat IgG binding and the reactivity with rat IgG subclasses and certain rat monoclonal antibodies is described. The potential application and limitations of the selected rat Fc receptor positive bacterial strain to immunoassays based on the specificity of rat monoclonal and polyclonal antibodies is discussed. PMID- 2978828 TI - Low responsiveness to MLM bacilli associated with an impaired macrophage function in CBA/J mice. AB - It has previously been reported that susceptibility to Mycobacterium lepraemurium (MLM) infection (subcutaneous inoculation) differs markedly between CBA/J and C57BL/6 mice. The high susceptibility of CBA/J mice was shown to be due to the generation of cyclophosphamide-sensitive, I-Jk-positive suppressor T cells. In the present study, we attempt to show functional differences between splenic macrophages of both strains of mice in terms of their phagocytic and antigen presenting activities against MLM bacilli. The results show that (a) the secondary MLM-induced T-cell proliferative response is dependent on Ia-positive antigen-presenting cells (APC), (b) macrophages of CBA/J mice are able to present antigens to F1 T cells, and yet (c) they show an extremely low phagocytic activity, (d) antigen presentation by APC of CBA/J mice lasted for a longer period than that of C57BL/6 mice after MLM infection. These results suggest that there is a functional difference in CBA/J and C57BL/6 APC, which could account for the phenotypic difference in immune responsiveness to MLM bacilli. PMID- 2978829 TI - Proliferative T cell clones directed at a human minor antigen. AB - Clones directed against a human minor antigen were isolated from an interleukin-2 dependent cytotoxic T cell line. As expected, some clones exhibiting cytotoxicity for HLA-identical minor antigen-positive cells were detected. In addition, some clones were detected which, though not cytotoxic, were able to proliferate in the primed lymphocyte typing (PLT) test in response to stimulation by cells from some HLA-identical sibs and certain DR 2+/ve unrelated cells. These clones expressed the Leu 3a cell surface antigen. DR 2 was one of the class II determinants expressed by the responding cell used to generate the cytotoxic T lymphocyte line. These results are consistent with the interpretation that helper T cells are able to recognize minor antigens in the context of self-class II HLA determinants and respond in PLT. PMID- 2978830 TI - Pregnancy outcome in human couples with recurrent spontaneous abortions: HLA antigen profiles; HLA antigen sharing; female serum MLR blocking factors; and paternal leukocyte immunization. AB - Critical features of the trophoblast for immune protection in the mother are: (1) its resistance to cytotoxic lymphocytes and antibodies; (2) it forms a physical barrier to immune effector cells, but not antibody, from reaching the fetus; (3) it signals the migration of suppressor and other functionally hyporesponsive lymphocytes into the uterine decidua and uterine lymphatics; (4) it promotes the production of maternal serum MLR (mixed lymphocyte reaction) blocking antibody with paternal antigen specificity. Some of these immunological features are lacking in women with recurrent abortions of immune etiology. Eleven women who aborted an additional time after immunization with paternal leukocytes were compared with 14 women who delivered infants at term post-immunization. It was found that those who aborted: (1) had HLA antigen profiles that did not differ significantly from those of control fertile couples or from observed antigen frequencies in North American Caucasians; (2) shared more HLA A, B, D/DR, and MT antigens with their spouses than controls; (3) were not more hyporesponsive in MLR to paternal antigens pre- and post-immunization when compared to controls; (4) failed to develop female serum MLR blocking factors post-immunization; (5) failed to develop humoral alloantibodies to B-cell alloantigens; (5) had lymphocytes in the uterine decidua mantling the conceptus and in the uterine lymphatics that were reactive/cytotoxic to paternal stimulating alloantigens. These results are in sharp contrast to the immunodynamics of peripheral blood leukocytes and decidual leukocytes to paternal alloantigens in women who delivered infants at term post-immunization. PMID- 2978831 TI - AIDS, public policy, and dentistry. PMID- 2978832 TI - Effect of phenytoin on the salivary composition in handicapped patients. PMID- 2978833 TI - Different fusimotor reflexes from the ipsi- and contralateral hind limbs of the cat assessed in the same primary muscle spindle afferents. AB - Experiments were performed in forty-five cats anaesthetized with alpha chloralose. The aim of the study was to investigate a sample of primary muscle spindle afferents from triceps muscle with respect to their fusimotor reflex control from ipsi- as well as contralateral hind limb. Primary muscle spindle afferents of the triceps surae muscle were recorded from the mean rate of firing and the modulation of the afferent response to sinusoidal stretching of the triceps surae muscle was determined. Test measurements were made during tonic stretch of the ipsilateral PBSt, contralateral PBSt, contralateral triceps muscle or during extension of the intact contralateral hind limb. Control measurements were made with ipsi- and contralateral PBSt as well as contralateral triceps muscles relaxed and with contralateral hind limb in resting position. The occurrence and types of fusimotor effects were assessed by comparing test to control responses. The main finding of the present investigation was the great variability in type and size of the fusimotor effects evoked by different ipsi- and contralateral reflex stimuli. Both ipsi- and contralateral stimulations gave rise to predominantly dynamic, predominantly static or mixed static and dynamic fusimotor reflexes. In the same preparation, a given reflex stimulus often caused different reflex responses in different triceps surae primary spindle afferents. In the same afferent unit, different reflex stimuli usually produced fusimotor effects which differed from each other in type and/or size. In general, contralateral whole limb extension and stretch of contralateral PBSt muscles were more potent as reflex stimuli than stretch of the ipsilateral PBSt muscle. Stretch of the contralateral triceps surae muscle was, but for a few afferent units, ineffective as reflexogenic stimulus. It is concluded that the individualized receptive profiles of the primary muscle spindle afferents, which have been postulated in earlier investigations where the effects of different stimuli have been investigated on different cell populations, still seems to hold good when the stimuli are tested on the same units. The individuality of the receptive profiles of gamma-motoneurones is discussed in relation to different motor control hypotheses. PMID- 2978836 TI - Present status of recombinant hepatitis B vaccine. PMID- 2978835 TI - Global strategy for prevention of hepatitis B. PMID- 2978834 TI - [Medroxyprogesterone acetate in fibrocystic disease of the breast in adolescents]. PMID- 2978837 TI - Rapid phage DNA isolation without the use of enzymes. PMID- 2978838 TI - A simple technique for preparing pure lambda DNA. PMID- 2978839 TI - Delayed-type hypersensitivity to allogeneic mouse epidermal cell antigens. III. Analysis of suppressor cells. AB - The specificity of suppressor T cells (Ts cells) induced by intravenous administration of allogeneic spleen cells was studied in mice using a delayed type hypersensitivity (DTH) assay. The DTH responses were induced by subcutaneous injection of allogeneic epidermal cells (ECs) and were assayed by footpad swelling. Afferent-phase Ts cells (Ts-aff cells) were transferred into the recipient mice before ECs immunization. Treatment with monoclonal anti-Thy-1.2 antibody and complement abolished the suppression by Ts-aff cells in the DTH response. The suppression induced by Ts-aff cells, however, was resistant to the treatment with monoclonal anti-I-A or anti-Lyt-2.2 antibody and complement. These results showed that Ts-aff cells were Lyt-2-, Ia- T cells. Efferent-phase Ts cells (Ts-eff cells) were transferred before challenge of the DTH assay. Phenotypic analysis of these Ts-eff cells showed them to be Lyt-2+, Ia- T cells. Studies using several strains of congenic mice revealed the antigen specificity of both Ts cell subsets. Adoptive transfer of Ts-eff cells required H-2 restriction, but Ts-aff cells did not. We also induced cognate suppression of the DTH responses to the alloantigens mediated by Ts-eff cells. PMID- 2978840 TI - Ly-5.185 molecule is associated with thymic maturation of lymphocytes but not with their cytotoxic activity. AB - The Ly-5 system is a composite family of molecules with different molecular weight. Each molecular form is characteristic of a different lymphoid population, in particular a 175,000- to 185,000-dalton doublet is peculiar of thymocytes and peripheral T cells. Athymic nude mice, however, lack the 185,000-dalton form on their lymphocytes. To investigate whether the expression of the 185,000 daltons requires the presence of thymus, we induced nude mice lymphocytes to differentiate into mature T cells in vivo by thymus grafting and, in vitro, into T cytotoxic cells (CTL) by cocultivation with alloantigen plus recombinant interleukin 2 (IL-2). The appearance of the 185,000-dalton form on lymphocytes of thymus-grafted nude mice but not on in vitro activated cytotoxic cells indicates an active role of thymus in the induction of this molecule and suggests that 185,000-dalton form could be a marker of intrathymic differentiation but not of the cytotoxic activity of T cells. PMID- 2978841 TI - Differential genetic requirements for in vivo and in vitro induction of T-killer and T-suppressor cells in the mutant H-2Kb system and the cross-reactivity of the T-killer clones. AB - Differences in the generation of anti-H-2Kb wild-type specific cytotoxic T lymphocytes (CTL) and specific suppressor T cells (SSTC) were investigated in H 2Kbm mutant mouse strains. To this end, optimal conditions for in vivo induction of highly active CTL in this mutant system were found and the T-cell origin of CTL and SSTC was confirmed using the anti-Thy-1.2 monoclonal antibody G4. Unlike the CTL, which were generated in vivo by any of the mutant strains tested (bm1, bm3, and bm4), the SSTC were only produced by bm3, whose H-2Kbm3 antigen, in contrast to the other H-2Kbm molecules, differs from wild type by serologically defined determinants. The high activity of anti-wild type bm4 CTL induced in vivo contrasted with a low activity of such CTL induced in mixed lymphocyte culture (MLC). This appeared to be the property of bm4 only, but not of bm1 or bm3, and it was reproduced in the reciprocal system B10 anti-bm4. CTL generation could be restored in the MLC by the addition of concanavalin A supernate or a mixture of bm4 and bm12 stimulator cells. Three of the six in vivo induced and in vitro propagated bm3 anti-B6 CTL clones demonstrated selective cross-reactivity to only one of the third-party H-2K molecules used, either Kk, Kd, or Kbm4. The present results indicate that (a) the SSTC and antibody recognize similar H-2Kb epitopes; (b) the H-2Kb epitopes recognized by the CTL and SSTC are not identical; (c) the genetic control of CTL generation in vivo is distinct from that in MLC, and (d) the affinities of antigen-specific receptors on T-cell clones of the same specificity may be different, leading to their individual cross-reactivity patterns. PMID- 2978842 TI - [Hypertension as a cardiovascular risk factor: physiopathologic mechanisms]. PMID- 2978843 TI - [Neurohumoral response to sublingual captopril and nifedipine in hypertensive emergencies and during the cold test. Contribution to the study of the interaction between the sympathetic and renin-angiotensin systems and atrial natriuretic factor]. PMID- 2978844 TI - [Evaluation of hypertension with the dynamic exercise test]. PMID- 2978845 TI - [Rheumatoid factor in secretory otitis media]. PMID- 2978846 TI - The rheumatology of Paget's disease. AB - Up to 60% of patients with symptomatic Paget's disease develop articular symptoms in its course. Effective therapy may depend on accurate localization of all symptoms and many patients will have an overlap of bone and articular symptoms which may prove impossible to separate. Over the past 10 years there have been major advances in drug therapy for Paget's disease but still the ideal agent is not currently available. Analgesics and non-steroidal anti-inflammatory drugs (NSAIDs) remain the basis of treatment for arthritis and there is no evidence yet that calcitonin or the diphosphonates will inhibit its development. Orthopaedic procedures, especially osteotomy for deformity and joint replacement may be indicated in selected cases. PMID- 2978847 TI - [Genetic polymorphism of human red cell phosphoglucomutase-1 subtypes in Chinese]. PMID- 2978849 TI - [The teaching of epidemiology in Italy]. PMID- 2978848 TI - Light and electron microscopic observations of muscle spindles in the rat tongue. PMID- 2978850 TI - [Continuing education in medicine: a dream, a mirage or a nemesis?]. PMID- 2978851 TI - [Is pleasure a risk factor?]. PMID- 2978852 TI - [Sentinel events in occupational environments: applications and developments]. PMID- 2978853 TI - [Isotretinoin (Roaccutan), a new teratogenic drug: which preventive strategies?]. PMID- 2978854 TI - [HIV-1 infection in Italy as of August 31, 1988]. PMID- 2978855 TI - [Mortality caused by tumors attributable to occupation exposure to asbestos amosite in a community]. PMID- 2978856 TI - [A system for screening and epidemiologic monitoring of HIV infection in drug addicts. A working model and analysis of data related to 1517 subjects from the Veronese area]. PMID- 2978857 TI - [Trends of the WHO]. PMID- 2978858 TI - [An interview with Dr. H. Nakajima, general director of the WHO. Interview by Giuseppe Lojacono]. PMID- 2978859 TI - [An interview with Dr. D. Poggiolini, director of the Pharmaceutical Service of the Health Ministry and the Drug Committee of the EEC. Interview by Giuseppe Lojacono]. PMID- 2978860 TI - [25th anniversary of the chair of pediatric stomatology]. PMID- 2978861 TI - [Dental health analysis and orthodontic anomalies in normal children and in children with hearing or sight loss]. PMID- 2978862 TI - Ofloxacin in the treatment of gonococcal and chlamydial urethritis. PMID- 2978864 TI - [Endodontic therapy in special patients]. PMID- 2978863 TI - [Fluoride release from restorative materials containing fluoride]. PMID- 2978865 TI - The organization and coherence of socioemotional, cognitive, and representational development: illustrations through a developmental psychopathology perspective on Down syndrome and child maltreatment. PMID- 2978866 TI - hst-1 transforming protein: expression in silkworm cells and characterization as a novel heparin-binding growth factor. AB - A protein encoded by an hst-1 transforming gene was expressed in silkworm-derived BmN cells and secreted into the culture medium using a recombinant baculovirus vector. The strong affinity for heparin of the secreted protein made it possible to purify the hst-1 protein to homogeneity in a two-step procedure. The purified hst-1 protein has a molecular weight of 18,000 and stimulates both DNA synthesis in NIH3T3 cells and human umbilical vein endothelial cell proliferation. In addition, morphological changes and anchorage-independent growth of NIH3T3 cells are induced by this product. These results show that the hst-1 transforming protein is a novel heparin-binding growth factor as predicted by nucleotide sequence analysis. PMID- 2978867 TI - DNA binding properties of murine p53. AB - We analysed the in vitro binding of p53 from normal (3T3) and from chemically transformed (Meth A) Balb/c mouse cells to double-stranded (ds-) DNA and to single-stranded (ss-) DNA by DNA-cellulose chromatography. We confirm previous findings that p53 in cellular extracts exhibits ds-DNA-binding activity (Lane and Gannon, 1983). In addition, we demonstrate that such p53 also binds to ss-DNA. Analyses with immunopurified p53 protein provide evidence that this DNA-binding activity is intrinsic to p53. DNA binding of p53 could not be inhibited by a monoclonal antibody specific for the C-terminal region. An N-terminal deletion mutant of p53 (Rovinski et al., 1987) exhibited similar DNA-binding properties as wild-type p53, indicating that the N-terminus also is dispensable for DNA binding. We further show a close correlation between the DNA-binding activity of p53 from 3T3 cells and its association with nuclear substructures. PMID- 2978868 TI - Cloning of the p53-dependent origin of cellular DNA replication. AB - We have recently reported that the c-myc protein may promote cellular DNA replication by binding to the origin of DNA replication (ori) and that an origin of human DNA replication which can autonomously replicate in human cells was cloned as a binding sequence of c-myc protein (Iguchi-Ariga et al., 1987). Here we report that cellular tumor antigen p53 may also participate in cellular DNA replication and another origin of DNA replication was cloned as a possible p53 binding sequence. The sequence could autonomously replicate in Raji cells which express p53 at a high level but not in HL-60 cells in which the coding gene for p53 is largely deleted. Little homology of the sequences was found between c-myc protein-binding ori and p53-binding ori. This suggests that c-myc protein and p53 may independently recognize different ori in chromosomal DNA. PMID- 2978869 TI - Specific interaction between a subset of the p53 protein family and heat shock proteins hsp72/hsc73 in a human osteosarcoma cell line. AB - We report here immunological evidence for the specific association between p53 and hsp72/hsc73 heat shock proteins in a human cell line derived from an osteosarcoma. The same association between p53 and hsp72/hsc73 was observed in HOS-TE85 clone 5 from which the HOS-SL cell line was derived. This association was indicated by the co-immunoprecipitation from HOS-SL of both p53 and hsp72/hsc73 proteins observed with either an anti-p53 monoclonal antibody (PAb421) or an antiserum specifically reacting with hsp72/hsc73 heat shock proteins. Furthermore, Western blot analysis allowed us to show that hsp72/hsc73 proteins did not share an epitope with p53, confirming that the co immunoprecipitation of p53 and hsp72/hsc73 was attributable to the physical association of the proteins. Data obtained from SDS-PAGE show that the HOS-SL cells expressed two forms of p53 with distinct molecular weights. Both forms contained several species with different isoelectric points ranging between pH 6.0 and 6.5. The data obtained from both 1D and 2D gel analyses consistently show that the p53 proteins involved in the association with hsp72/hsc73 were mainly the species that migrated with the slower mobility in the SDS dimension. The possibility is discussed that the HOS-SL p53 variant forming a complex with hsp72/hsc73 contains an activating mutation for transformation. PMID- 2978870 TI - [Risk of infections transmission during stomatological treatment]. PMID- 2978871 TI - [Oral status of children with chronic renal insufficiency]. PMID- 2978872 TI - [Neuropsychological deterioration in Huntington chorea]. PMID- 2978873 TI - [Cerebrovascular disorders as a presentation form of hypertrophic myocardiopathy]. PMID- 2978875 TI - Improved left ventricular function after short-term treatment with fructose-1,6 diphosphate: echocardiographic study in chronic ischemic heart disease and idiopathic dilated cardiomyopathy. AB - The effect of fructose-1,6-diphosphate (FDP) on left ventricular function was assessed in seven patients with chronic ischemic heart disease and eight patients with idiopathic dilated cardiomyopathy. In a crossover study design each patient received 10 gm of FDP or saline placebo intravenously for three days. An M-mode echocardiographic assessment of left ventricular (LV) function was made before and after each treatment period. After FDP treatment, LV end-diastolic and systolic dimensions showed a 6% reduction (P less than 0.01), while peak lengthening rate of LV dimension in diastole and peak shortening rate of LV dimension in systole increased 17% and 10%, respectively (P less than 0.05). There was evidence that FDP was more effective in the patients with ischemic heart disease than in the patients with cardiomyopathy. PMID- 2978874 TI - Effects of glibenclamide on serum lipids, lipoproteins, thromboxane, beta thromboglobulin, and prostacyclin in non-insulin-dependent diabetes mellitus. AB - A study was conducted to determine the effects of glibenclamide on serum lipoproteins, apolipoproteins, thromboxane (TXA2), prostacyclin (PGI2), and beta thromboglobulin (B-TGL) in patients with newly diagnosed non-insulin-dependent diabetes mellitus (NIDDM). In 20 NIDDM patients, aged 34 to 67 (mean, 53.6) years, without clinical signs of atherosclerotic disease and whose blood sugar level was over 140 mg/dl after four weeks of dietary treatment, fasting blood samples were taken before the beginning of the trial, after four weeks of dietary treatment, and after four and eight weeks of combined dietary and glibenclamide treatment. Pretrial levels of total serum cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) in the diabetic patients did not differ from those in nondiabetic controls, whereas high-density lipoprotein cholesterol (HDL-C) levels and the percentage of TC bound to HDL (HDL-C%) were significantly lower in the patients than in controls. After combined dietary and glibenclamide treatment and the normalization of blood sugar, both HDL-C (mg/dl) levels and HDL-C% levels increased significantly. TC, TG, and LDL-C levels decreased. Levels of apolipoproteins A1 and A2 rose and apolipoprotein B fell, but differences were not significant. TXB2 and 6-keto-PGF1-alpha (the inert metabolites of TXA2 and PGI2) and B-TGL were determined by radioimmunoassay. TXB2 and B-TGL levels decreased significantly after glibenclamide administration, indicating attenuation of platelet aggregation. No changes in PGI2 were observed. The results demonstrate the favorable effect of glibenclamide on lipoproteins and apolipoproteins in NIDDM patients, especially in increasing HDL-C levels and HDL C%, and in attenuating platelet aggregation as indicated by reduction of TXB2 and B-TGL. PMID- 2978876 TI - The relationship of immunological responsiveness to suppression of cellular responses by cyclosporine in vitro. PMID- 2978878 TI - The role of suppressor cells in the blood transfusion phenomenon. PMID- 2978877 TI - Cyclosporine inhibits nonspecific transfusion-induced immunity in dogs. PMID- 2978879 TI - Identification of cell population mediating suppression after blood transfusion. PMID- 2978880 TI - The transfusion effect in dogs: no indication for the presence of transfusion induced suppressor cells in the spleen. PMID- 2978882 TI - Modulation of T cell subsets in immune responses to alloantigens by allo-specific blood transfusion. PMID- 2978881 TI - Blood transfusion induces alterations in the circulation of immunoglobulin secreting cells and activates helper T cells for Ig synthesis. PMID- 2978883 TI - The clinical significance of the autologous mixed lymphocyte reaction in transplantation. PMID- 2978884 TI - In vitro humoral responses of renal allograft recipients. PMID- 2978885 TI - In vivo-in vitro correlates of human rejection using flow cytometry with two color fluorescence on peripheral blood mononuclear cells. PMID- 2978887 TI - MLR suppression and MLR-suppressor activity induced by thoracic duct drainage prior to transplantation. PMID- 2978886 TI - Immunogenicity and viability of alpha-class II immunotoxin-treated pig islet cells. PMID- 2978888 TI - Stopping prednisone in kidney transplant recipients taking cyclosporine: safety conditions. PMID- 2978889 TI - Percutaneous transluminal angioplasty: the procedure of choice for renal allograft artery stenosis. PMID- 2978890 TI - Three year experience of donor specific transfusion and concomitant cyclosporine A. PMID- 2978892 TI - Kinetics of antidonor mixed lymphocyte culture responses in donor specific transfusions plus azathioprine-treated patients. PMID- 2978891 TI - Optimization of donor specific blood transfusion in kidney transplantation. AB - 1. One hundred milliliters of stored whole blood DST, three times at weekly intervals is a practical, less immunizing and effective approach to enhance graft survival in recipients of a living-related donor kidney. 2. This protocol could also be used in sibling donor/recipient pairs who do not share a haplotype as well as in those who share two haplotypes to enhance graft survival. 3. The use of a short course of Cs (6 mg/kg/d for three weeks) along with DST appears to reduce the sensitization rate even lower. The dose of Cs used in this study produced no clinically significant adverse reactions, whereas Aza (1 mg/kg/d) often produces leukopenia. 4. DST produces significantly suppressed donor specific MLC responses in the early post-DST period; however, it increases the response at a later time. DST modulates immune responses in such a way that secondary responses upon grafting are more readily reversible by immunosuppressive agents. PMID- 2978893 TI - Related donor kidney transplantation is the best form of treatment for uremia. PMID- 2978894 TI - Immunoprophylaxis of hepatitis B virus reinfection in recipients of human liver allografts. PMID- 2978895 TI - Predictors of rejection in cyclosporine-treated cardiac allograft recipients. PMID- 2978896 TI - Role of suppressor cells and thymic hormone function in the early and late phases of murine graft-versus-host reactions. PMID- 2978897 TI - Induction and suppression of graft-versus-host reactivity in B6.C-H-2 mutant mice. PMID- 2978898 TI - [D-penicillamine and cutaneous lesions in Wilson's disease]. PMID- 2978899 TI - Stress-induced changes in plasma concentrations of immunoreactive beta-endorphin and cortisol in response to routine surgical procedures in lambs. AB - Following four different surgical procedures in lambs 3-5 weeks old, plasma immunoreactive beta-endorphin (beta-EP) and cortisol were assayed at 15 min and 24 h as determinants of post-operative stress. A threefold increase in mean plasma beta-EP levels occurred 15 min after tail docking, and a maximal eight- to tenfold increase occurred in response to castration and/or mulesing with tail docking. Significant increments in mean plasma cortisol levels followed these surgical procedures with the maximal response 15 min after mulesing plus castration with tail docking. The physiologically active 'free' cortisol in plasma represents about 25% of the cortisol, as measured, and the two are highly correlated. At 24 h, beta-EP levels in all treated groups were similar to controls, although a small elevation in cortisol levels was still present in the lambs subjected to mulesing. Ultrafiltration of plasma extracts showed that peak beta-EP levels contained about 40% immunoreactivity from low molecular weight species (mol. wt less than 10,000). By specific radioimmunoassay and reverse phase high-performance liquid chromatography this comprised about 75% beta-EP1 31, the most potent analgesic endorphin, 10% beta-EP1-27, and 15% alpha-N-acetyl beta-EP. Increased beta-EP1-31 levels may modulate post-operative pain in lambs. PMID- 2978900 TI - Effect of adding ketotifen to ipratropium bromide in the treatment of nocturnal asthma. PMID- 2978901 TI - Cyclosporine sensitivity of human alloreactive T lymphocytes depends on the strength of the allogeneic stimulus. PMID- 2978903 TI - The alloreactive potential of T-like cells from athymic nude rats (rnu/rnu). PMID- 2978902 TI - Amplification of in vitro tissue-associated (mixed lymphocyte kidney culture) immunity by allograft rejection. PMID- 2978904 TI - Suppression of murine alloimmune cytolytic T lymphocyte development by spleen cells incubated with staphylococcal enterotoxin B in vitro. PMID- 2978905 TI - Limiting dilution analysis of human alloantigen-reactive helper T lymphocytes. PMID- 2978906 TI - Requirements for primary T cell activation. PMID- 2978907 TI - Intrinsic regulation programs for responses to alloantigens within monoclonal human helper T lymphocyte populations. PMID- 2978908 TI - Functional analysis of TDTH and Tc cells infiltrating primary or secondary allografts. PMID- 2978909 TI - T helper cells infiltrating kidney allografts: frequency and functional characterization. PMID- 2978911 TI - Spontaneous acceptance of spleen allografts: role of T suppressor cells at the graft site. PMID- 2978910 TI - Nonspecific tissue destruction as a consequence of cytotoxic T lymphocyte interaction with antigen-specific target cells. PMID- 2978912 TI - Induction of suppressor cells by ultraviolet-irradiated DST in monkey skin allografts. PMID- 2978913 TI - The nature of T cell repertoire modification in neonatal tolerance. PMID- 2978914 TI - Mechanisms of immunologic tolerance following total lymphoid irradiation in the baboon. PMID- 2978915 TI - Specific unresponsiveness in rats with prolonged allograft survival is dependent upon the graft and suppressor T cells. PMID- 2978916 TI - Immune responsiveness following skin grafting in cyclosporine-treated rats. PMID- 2978917 TI - Tempo of induction of W3/25+ suppressor cells in cyclosporine-treated rat cardiac allograft recipients. PMID- 2978918 TI - Suppression of allograft rejection in the rat by pretreatment with cells of the T helper, but not T-cytotoxic suppressor phenotype. PMID- 2978919 TI - Alloreactive suppressor T cells. PMID- 2978920 TI - Contra-IL 2 activity of a lymphokine produced by a hybrid cell line of ALS induced suppressor T cells. PMID- 2978922 TI - Blood transfusion plus allograft--but not blood transfusion alone--induce IL 2 producing suppressor cells in Lew-1A recipients of LEW-1W heart allograft. PMID- 2978921 TI - Cloned natural suppressor cells derived from the neonatal spleen: in vitro action and lineage. PMID- 2978923 TI - Veto cells are not responsible for alloimmunization-induced abrogation of lethal graft-versus-host reaction. PMID- 2978924 TI - W3/25+ and OX8+ T suppressor cells are required to maintain transplantation tolerance. PMID- 2978925 TI - Regulation of neonatal transplantation tolerance induction by natural suppressor and contrasuppressor cells. PMID- 2978926 TI - Anti-interleukin-2 receptor (IL-2R) antibody against rejection of organ grafts. PMID- 2978927 TI - Ia-positive donor-specific bone marrow cells prolong renal allografts in dogs treated with fractionated lymphoid irradiation. PMID- 2978928 TI - Comparison of clinical response of DR-matched, MLC-compatible cadaver renal allografts and those from HLA-identical related donors. PMID- 2978929 TI - Donor-specific T cell alloreactivity in relation to 1 haplotype-mismatched blood transfusion prior to living-donor renal transplantation. PMID- 2978930 TI - The effect of donor-specific blood transfusion on in vitro alloreactive precursor cell frequencies. PMID- 2978931 TI - Functional heterogeneity of HLA-class II determinants: the role of HLA-DQ as a modulator of cell-mediated responses. PMID- 2978932 TI - Mechanism of specific transplantation tolerance induction by vascular allografts across a class I difference in miniature swine. PMID- 2978933 TI - Response to Mlsa and H-2 cross-reactivity of long-term T cell lines generated between Mls congenic mice. PMID- 2978934 TI - T cell presensitization in renal transplantation. PMID- 2978935 TI - [Modification of the pain threshold and beta-endorphin level in heroin addicts undergoing withdrawal]. PMID- 2978936 TI - [Diagnosis and management of ectopic pregnancies]. PMID- 2978937 TI - Adverse drug reactions: reporting and evaluating cutaneous reactions. PMID- 2978938 TI - Lichen planus and related conditions. PMID- 2978939 TI - Workers' compensation. PMID- 2978940 TI - Molecular genetics of yeast: establishing the relationship between a gene and its function. PMID- 2978941 TI - [Diagnostic laparoscopy. Experience with 1,287 cases]. PMID- 2978942 TI - Impaired induction of delayed hypersensitivity following anterior chamber inoculation of alloantigens. AB - BALB/c mice that receive allogeneic P815 tumor cells (DBA/2 origin) into the anterior chamber of the eye (intracamerally, i.c.) fail to develop DBA/2-specific delayed hypersensitivity (DH). We have previously shown that this failure results, in part, from the generation of efferent T suppressor cells. To determine whether DH effector cells are even activated in these mice, a local adoptive transfer assay for DH was developed: Mixtures of responder lymphocytes and irradiated stimulator cells were injected into the pinnae of normal mice, syngeneic with the responders. In this assay, spleen cells from i.c. injected donors failed to evoke a local DH. Moreover, when spleen cells from i.c. injected donors were mixed with specifically immune BALB/c anti-DBA/2 lymphoid cells and similarly assayed, DH was effectively suppressed. Spleen cells from i.c. injected mice, depleted of Lyt-2+ suppressor cells, remained unresponsive in this assay, even if before injection they had been exposed to 5 days of in vitro culture with DBA/2 alloantigens. We conclude that anterior chamber inoculation of P815 cells in BALB/c mice fails to activate DH effector cells. Therefore, the inability of i.c. injected mice to mount DH responses results not only from efferent suppression but also from selective impairment of DH induction. The latter effect may be due to the activation of an afferent suppressor mechanism that acts selectively on T cell help for DH but that has no effect on help for B cells or cytotoxic T cell precursors. PMID- 2978943 TI - Genetic restrictions and cellular interactions in the induction of anterior chamber associated immune deviation (ACAID). AB - The injection of antigen into the anterior chamber (AC) of the eye results in a pattern of systemic immune responses termed anterior chamber associated immune deviation (ACAID). ACAID is characterized by down-regulation of delayed-type hypersensitivity (DTH) by T suppressor cells with normal antibody production and cytotoxic T cell responses. To better understand the mechanisms of ACAID, we studied the genetic restrictions and intraocular T cell interactions in TNP ACAID. We report that (a) antigen is not reprocessed by the eye for the induction of suppression in TNP-ACAID, (b) the T cells injected into the AC and the recipient animal must match at genes in the immunoglobulin heavy chain (IgH) region of chromosome XII for TNP-ACAID to be induced, (c) TNP-coupled Lyt-1+, I J+ T cells present antigen to viable T cells within the eye to induce systemic suppression, (d) although there is an IgH restriction between the T cells injected into the AC and the recipient, the interaction between the T cell antigen-presenting cell and the Tsi-cells in the AC inoculum is restricted to the I-J portion of the major histocompatibility complex, and (e) the two Tsi-cells that initiate the suppressor cascade have different antigen presentation requirements. These results further emphasize the complex interaction between T cells, which serves to control immune reactions in the eye. PMID- 2978944 TI - Serodiagnosis of the acquired immune deficiency syndrome by enzyme-linked immunosorbent assay compared to cellular immunologic parameters in African AIDS patients and controls. AB - The sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA), absolute numbers of T-helper cells, and T-helper: T-suppressor cell ratios were compared in asymptomatic controls and IgG Western blot-confirmed patients with the acquired immune deficiency syndrome (AIDS) in Kinshasa, Zaire, between August 1984 and May 1985. Two hundred sixteen (97.7%) of 221 IgG Western blot-positive AIDS patients and 4 of 97 (4%) controls were ELISA-positive, 3 of whom were Western blot-positive. The sensitivity and specificity of the ELISA was 97.7% and 99.0%, respectively, compared to Western blot results. Detection of the human immune deficiency virus using absolute number of T-helper cells (less than 400 cells/mm3) was as sensitive (98.2%), but less specific (90.7%). A T-helper: T suppressor ratio of less than 0.9, had a sensitivity of 97.3%, and specificity of 94.8%. The ELISA test had the highest predictive value and greatest utility in an African clinical setting for detecting HIV infected patients where a wide range of other immunocompromising diseases are seen. PMID- 2978945 TI - [Diphosphonates and primary hyperparathyroidism]. PMID- 2978946 TI - Cellular immunoregulation of acute pulmonary inflammation in strain 2 guinea pigs. AB - Subclasses of lung immunoregulatory T cells were analyzed during acute pulmonary inflammation in strain 2 guinea pigs and compared with T cell subpopulations in the peripheral circulation. Immunized animals were aerosol-challenged with specific antigen and sacrificed at 12-, 24-, 48-, 72-, and 96-hours postchallenge. Mononuclear cells, isolated from peripheral blood and bronchoalveolar lavage, were enriched for T cells. The percentage of helper T cells, as well as antigen-specific blastogenesis, in recovered pulmonary T cells exhibited maximal values at 12-, 24-, and 96-hours postchallenge. In contrast, the presence of suppressor T cells correlated with decreased blastogenesis and antigen-specific suppression in isolated lung cells at 72-hours postchallenge. Since changes in pulmonary cells did not correlate with those found in the peripheral circulation, immunoregulatory events in these two compartments may be distinct. These results indicate that the proportions of lung T cell subclasses, as well as their in vitro functional activity, are altered over the course of pulmonary disease. Such changes in immunoregulatory cell populations may be important in the mediation of disease pathogenesis. PMID- 2978947 TI - A novel suppressor cell develops in uterine decidua in response to fetal trophoblast-type cells. AB - In both murine and human pregnancy, a novel population of small lymphocytic cells with suppressive activity accumulates in the uterine decidua. In the mouse, these suppressor cells lack T cell markers, release an interleukin-2 blocking factor, are associated with cells with cytoplasmic granules, and are deficient in the decidua of mice showing increased abortion rates with loss of resistance to attack by maternal antipaternal transplantation immunity. Small granulated cells with immunosuppressive activity do not arise as a result of decidualization alone but can be induced using trophoblast-type cells and supernatants conditioned by trophoblast-type cells. Possible mechanisms of this induction and the importance of trophoblast-decidual interaction in generating localized immunosuppressive buffer zone (chemical barrier) about the implanted conceptus are discussed. PMID- 2978948 TI - Lipoprotein(a): a genetically-determined lipoprotein variant in which a protein of the plasminogen family is linked to APO B100. PMID- 2978949 TI - Structure of ferrichrome-type siderophores with dissimilar N delta-acyl groups: asperchrome B1, B2, B3, D1, D2 and D3. AB - Asperchromes are a series of iron-chelating compounds which contain a cyclic hexapeptide backbone as in ferrichrome siderophores and differ from the latter in having heterogenous acyl groups in the ornithine side chains. The molecular structures of the asperchrome B and D series have been determined by 1H- and 13C NMR spectroscopy; single-crystal X-ray diffraction was used to determine the detailed structural features of asperchrome B1 and asperchrome D1. Asperchrome B1 crystallizes in the triclinic space group P1 with a = 1.3143(5) nm, b = 1.2200(5) nm, c = 0.8949(3) nm, alpha = 105.17(4) degrees, beta = 94.03(3) degrees, gamma = 109.65(3) degrees, V = 1.2843 nm3, Z = 1, rho chi = 1.446 g cm-3. Final R = 0.054 for 4625 reflections measured at 138 K using MoK alpha. Asperchrome D1 crystallizes in the monoclinic space group P2(1) with a = 1.2248(11) nm, b = 1.3795(9) nm, c = 1.3644(6) nm, beta = 93.24(6) degrees, V = 2.3016 nm3, Z = 2, rho chi = 1.418 g cm-3. Final R = 0.110 for 3180 reflections measured at 138 K using MoK alpha radiation. The conformation of the molecular backbone and iron coordination geometry in both asperchrome B1 and D1 compare well with those observed in other known ferrichrome siderophores. The differences in the acyl groups are illustrated and the structural results are correlated with their iron transport properties. PMID- 2978950 TI - Influence of water activity and temperature on the accumulation of zearalenone in corn. AB - The influence of water activity (aw) and temperature on the zearalenone biosynthesis in corn has been examined. Viable corn kernels were conditioned at different values of water activity (0.90, 0.95 and 0.97), inoculated with Fusarium graminearum and incubated at different temperatures. Zearalenone was determined at selected times. For the strain used, a constant temperature of 25 degrees C resulted more favorable than 15 degrees C, 20 degrees C and the combination of two weeks at 25 degrees C followed by incubation at 15 degrees C shown by other authors to increase the zearalenone accumulation. Both F. graminearum growth and zearalenone production at 25 degrees C were inhibited at water activity 0.90. At short incubation times, toxin accumulation was greater at water activity 0.97 than at water activity 0.95 (25 degrees C). This relation was inverted at longer periods of incubation. A combined effect of water activity and temperature on the zearalenone accumulation was observed. PMID- 2978951 TI - Inhibitory effects of selected Turkish spices and oregano components on some foodborne fungi. AB - The inhibitory effects of 10 selected Turkish spices, oregano essential oil, thymol and carvacrol towards growth of 9 foodborne fungi were investigated in culture media with pH 3.5 and 5.5. The antifungal effects of sodium chloride, sorbic acid and sodium benzoate and the combined use of oregano with sodium chloride were also tested under the same conditions for comparison. Of the spices tested, only sodium chloride were also tested under the same conditions for comparison. Of the spices tested, only oregano at 1.0, 1.5, 2.0% (w/v) levels showed effect on all fungi. 8% (w/v) sodium chloride was less effective than oregano. Oregano essential oil, thymol or carvacrol at concentrations of 0.025% and 0.05% completely inhibited the growth of all fungi, showing greater inhibition than sorbic acid at the same concentrations. The combined use of oregano and sodium chloride exhibited a synergistic antifungal effect. PMID- 2978952 TI - [Serum complement activation pathways and circulating anti-streptolysin-0 antibodies: diagnostic value in glomerulonephritis in children]. PMID- 2978953 TI - Histopathologic reactions to chemotherapeutic agents. AB - This chapter represents the first effort to characterize the cutaneous histologic changes that occur in response to chemotherapy. In preparing this chapter it became increasingly evident that there are significant gaps in our knowledge and that many of the known clinical reactions have not had careful histologic study. The readers of this chapter are encouraged to report adverse reactions they have encountered and include complete histologic descriptions of biopsied material. PMID- 2978954 TI - [Evidence for the existence of a positive feedback of the degree of expansion of the circulating volume, atrial distension and atrial natriuretic peptide in dialyzed uremic patients]. PMID- 2978955 TI - Possible chronological events leading to pancreatic beta cell damage in type I diabetes mellitus. PMID- 2978956 TI - Ferricrocin functions as the main intracellular iron-storage compound in mycelia of Neurospora crassa. AB - Neurospora crassa produces several structurally distinct siderophores: coprogen, ferricrocin, ferrichrome C and some minor unknown compounds. Under conditions of iron starvation, desferricoprogen is the major extracellular siderophore whereas desferriferricrocin and desferriferichrome C are predominantly found intracellularly. Mossbauer spectroscopic analyses revealed that coprogen-bound iron is rapidly released after uptake in mycelia of the wild-type N. crassa 74A. The major intracellular target of iron distribution is desferriferricrocin. No ferritin-like iron pools could be detected. Ferricrocin functions as the main intracellular iron-storage peptide in mycelia of N. crassa. After uptake of ferricrocin in both the wild-type N. crassa 74A and the siderophore-free mutant N. crassa arg-5 ota aga, surprisingly little metabolization (11%) could be observed. Since ferricrocin is the main iron-storage compound in spores of N. crassa, we suggest that ferricrocin is stored in mycelia for inclusion into conidiospores. PMID- 2978957 TI - N alpha-dimethylcoprogens. Three novel trihydroxamate siderophores from pathogenic fungi. AB - Three novel siderophores have been isolated from a highly pathogenic strain of Alternaria longipes (ATCC 26293). The compounds are N alpha-dimethylated analogs of coprogen, neocoprogen I and isoneocoprogen I. Structures of the compounds have been determined by 1H- and 13C-NMR, fast-atom-bombardment (FAB) mass spectroscopy and partial hydrolysis. One of the new compounds, N alpha-dimethylcoprogen, is also produced, as the major siderophore, in another fungus, Fusarium dimerum. PMID- 2978958 TI - Characterization of ferrioxamine E as the principal siderophore of Erwinia herbicola (Enterobacter agglomerans). AB - Several strains of the enterobacterial group Erwinia herbicola (Enterobacter agglomerans) were screened for siderophore production. After 3 days of growth in a low-iron medium, all strains studied produced hydroxamate siderophores. The retention values of the main siderophore during thin-layer chromatography on silica gel plates and on HPLC reversed-phase columns were identical with those of an authentic sample of ferrioxamine E (norcardamine). Gas-chromatographic analysis of the HI hydrolyzate yielded succinic acid and 1,5-diaminopentane in equimolar amounts; fast-atom-bombardment (FAB) mass spectroscopy showed a molecular mass of 653 Da. Iron from 55Fe-labelled ferrioxamine E was well taken up by iron-starved cells of E. herbicola (Km = 0.1 microM, Vmax = 8 pmol mg-1 min 1). However, besides ferrioxamine E (100%), several exogenous siderophores such as enterobactin (94.5%), ferric citrate (78.5%), coprogen (63.5%) and ferrichrome (17.5%) served as siderophores, suggesting the presence of multiple siderophore receptors in the outer membrane of E. herbicola. PMID- 2978959 TI - High-performance liquid chromatography of siderophores from fungi. AB - A reversed-phase HPLC separation of iron(III) chelates of 16 representative fungal siderophores including ferrichromes, coprogens and triacetylfusarinine C was established in order to investigate siderophore production of fungi. For comparison purposes, the widely used bacterial siderophore ferrioxamine B was included. Culture filtrates of the fungi Penicillium resticulosum, Fusarium dimerum, Aspergillus fumigatus and Neurospora crassa were quantitatively analyzed for the presence of known and unknown siderophores after growth in low-iron culture media and adsorption on XAD-2 columns using this HPLC separation system. Photodiode array detection allowed the distinction between siderophores and non siderophores. According to their ultraviolet/visible spectra, a further classification of the siderophores into four types due to the number of anhydromevalonic acid residues per molecule (0-3) was possible. PMID- 2978960 TI - [Abdominal "mini-lipectomy" associated with lipo-aspiration in the treatment of deformities of the abdominal wall]. PMID- 2978961 TI - Megakaryoblastic leukemia and Down's syndrome: a review. AB - Megakaryoblastic leukemia and transient leukemia in Down's syndrome have been reviewed using case reports from the literature and our own experience at the Hospital for Sick Children. The following conclusions have been reached: (1) approximately 20% of leukemia (excluding transient leukemia) in Down's syndrome is acute megakaryoblastic leukemia; (2) approximately 20% of all leukemia in Down's syndrome is transient leukemia; (3) transient leukemia in Down's syndrome is acute megakaryoblastic leukemia; (4) recurrence of acute megakaryoblastic leukemia occurs in 20% of the cases of transient leukemia; and (5) the incidence of acute megakaryoblastic leukemia in Down's syndrome is estimated to be 400 times that in normal children. These observations suggest that a specific form of leukemia, namely acute megakaryoblastic leukemia, has a remarkable association with Down's syndrome. PMID- 2978962 TI - [Down's syndrome: obstetrical aspects]. PMID- 2978963 TI - Subunit, recombinant and synthetic hepatitis B vaccines. AB - The failure to propagate hepatitis B virus in vitro has prevented the development of vaccines from virus grown in cell cultures. Alternative approaches including the use of surface antigen purified from plasma, subunit polypeptide vaccines, vaccines produced by recombinant DNA technology, expression by a recombinant vaccinia virus and the chemical synthesis of small peptides representing specific regions of surface antigens are reviewed in this article. PMID- 2978964 TI - Human cord blood monocytes: binding and ingesting capacity mediated by Fc and C3b receptors. AB - Human cord blood monocytes have been compared to monocytes from adults. Our results show that unstimulated cord blood monocytes are as effective as monocytes from adults with regard to their binding and ingesting capacity mediated by Fc and C3b receptors. Furthermore, when stimulated with PMA (phorbol-myristate acetate), the C3b receptor-mediated phagocytosis in cord blood monocytes was enhanced to the same extent as in monocytes from adults. These findings show that Fc and C3b receptor functions in human monocytes are fully developed at birth and cannot explain the increased susceptibility to infections seen in the neonate. PMID- 2978965 TI - T-lymphoblasts with erythropoietic helper function in acute T-cell leukemia. AB - A patient with acute T-lymphoblastic leukemia was found to maintain a normal hemoglobin concentration both at presentation and preterminally several months later, despite a replaced bone marrow and over 80% circulating lymphoblasts on both occasions. Cell surface marker analysis demonstrated the T-lymphoblasts both at presentation and preterminally to belong to the T-helper subpopulation. In vitro culture studies demonstrated that the patient's T-lymphoblasts, as well as conditioned medium derived from these lymphoblasts, significantly stimulated normal bone marrow erythroid colony growth (CFU-E). These findings suggest that in this patient the preservation of erythropoiesis resulted from a helper effect exerted by his T-lymphoblasts. PMID- 2978966 TI - The involvement of atriopeptins in blood pressure regulation. AB - The atriopeptins are newly discovered cardiac-derived peptides whose observed actions suggest a role in volume homeostasis and blood pressure regulation. Studies in animal models are underway to pinpoint pathogenetic mechanisms involved in the evolution of hypertension, some of which may well be shared by humans with "essential" hypertension. Preliminary observations indicate that circulating atriopeptin levels are altered in human disease. It is anticipated that exogenously administered atriopeptin may be a helpful pharmacological tool in the management of patients with volume overload and hypertension. PMID- 2978967 TI - Percutaneous transluminal angioplasty for renovascular hypertension in children. AB - Percutaneous transluminal angioplasty (PTA) has been developed over the past 8 years as an alternative to reconstructive surgery for renovascular hypertension. We report three cases and review the use of PTA in children with renal artery stenosis. At least 37 cases of PTA have been reported in patients whose ages ranged from 1.3 to 17 years (mean 10 years). Of these, 10 had fibromuscular dysplasia; 13 unspecified unilateral renal artery stenosis; 4 bilateral stenosis; 4 neurofibromatosis; 4 renal transplant; 1 atherosclerosis; and 1 postsurgical stenosis. Nine of 10 patients with fibromuscular dysplasia were cured and 3 of 4 with renal transplant arterial stenosis were cured or improved. There were 11 failures of PTA, including all 4 patients with neurofibromatosis and 1 with transplant arterial stenosis. We conclude that PTA is the treatment of choice for children with hypertension due to fibromuscular dysplasia and should be attempted for stenosis of the transplanted renal artery. Other lesions resulting in renal artery stenosis may not be as amenable to dilation and should be considered on an individual basis. PMID- 2978968 TI - The syndrome of hypertension and hyperkalaemia with normal glomerular function (Gordon's syndrome). A pathophysiological study. AB - A 14-year-old boy with the syndrome of hypertension and hyperkalaemia with normal glomerular filtration rate (Gordon's syndrome) is described. The patient's clinical symptoms consisted of periodic paralysis, slight metabolic acidosis of the proximal type and hypercalciuria. Prostaglandin excretion was normal. Infusion of atrial natriuretic peptide had no effect on electrolyte excretion or glomerular function although a normal increase in cyclic guanosine monophosphate was demonstrated in plasma and urine. This lack of sensitivity to atrial natriuretic peptide offers a new pathophysiological concept in this syndrome. Treatment with hydrochlorothiazide was successful in this case. PMID- 2978969 TI - [Postoperative necrotizing infections of the abdominal wall]. PMID- 2978970 TI - Clinical, histologic, and immunopathologic comparison of pemphigus vulgaris and pemphigus foliaceus. PMID- 2978971 TI - Uptake and destruction of 125I-CSF-1 by peritoneal exudate macrophages. AB - The binding and uptake of the colony-stimulating factor CSF-1 by peritoneal exudate macrophages (PEM) from lipopolysaccharide insensitive C3H/HeJ mice was examined at 2 degrees C, and at 37 degrees C. At 2 degrees C, 125I-CSF-1 was bound irreversibly to the cell surface. At 37 degrees C, 90% of the cell surface associated 125I-CSF-1 was rapidly internalized and subsequently degraded and the remaining 10% dissociated as intact 125I-CSF-1. Thus classical equilibrium or steady state methods could not be used to quantitatively analyze ligand-cell interactions at either temperature, and alternative approaches were developed. At 2 degrees C, the equilibrium constant (Kd less than or equal to 10(-13) M) was derived from estimates of the rate constants for the binding (kon congruent to 8 x 10(5) M-1 s-1) and dissociation (koff less than or equal to 2 x 10(-7) s-1) reactions. At 37 degrees C, the processes of dissociation and internalization of bound ligand were kinetically competitive, and the data was formally treated as a system of competing first order reactions, yielding first order rate constants for dissociation, koff = 0.7 min-1 (t1/2 = 10 min) and internalization, kin = 0.07 min-1 (t 1/2 = 1 min). Approximately 15 min after internalization, low molecular weight 125I-labeled degradation products began to appear in the medium. Release of this degraded 125I-CSF-1 was kinetically first order over three half lives (Kd = 4.3 x 10(-2) min-1, t1/2 = 16 min). Thus CSF-1 binds to a single class of receptors on PEM, is internalized with a single rate limiting step, and is rapidly destroyed without segregation into more slowly degrading intracellular compartments. PMID- 2978973 TI - [Infective endocarditis in dental practice]. PMID- 2978972 TI - Quantification of messenger ribonucleic acid for atrial natriuretic factor in atria and ventricles of Dahl salt-sensitive and salt-resistant rats. AB - The levels of atrial natriuretic factor (ANF) and the mRNA for ANF were measured in the left ventricles of Dahl salt-sensitive (S) and salt-resistant (R) rats. ANF and ANF mRNA were both much higher in ventricular tissue of newborn rats of both strains compared to young adults, which represents the normal developmental pattern. There was no strain difference between S and R when the rats were young (1.5 months of age), but in older animals (8.5 months of age), when S rats were markedly hypertensive, there was a 5- to 10-fold increase in both left ventricular ANF and left ventricular ANF mRNA in S, but not R, rats. Atrial ANF mRNA was not similarly increased in hypertensive S rats. The ANF levels present in ventricles could not be accounted for by contamination with plasma ANF. Moreover, HPLC analysis of the forms of ANF in ventricles of newborn and hypertensive S rats showed that immunoreactive ANF in ventricles was present mainly in the same precursor form found in atria and not the shorter peptide form found in plasma. Northern blot analysis showed that ANF mRNA for atria and ventricles were the same size. It is concluded that in the S rat the heart left ventricle responds to hypertension by increasing production and storage of ANF. PMID- 2978974 TI - [Exaggerated gag reflex and its management by dentist]. PMID- 2978975 TI - [Agranulocytosis and dental practice]. PMID- 2978976 TI - [Hemophilic patients. Treatment protocol in the dental office]. AB - Hemophilia is an inherited hemorrhagic disease which is due to the insufficiency of Factor VIII, or Factor IX, or Factor XI. Hemophilia patients are regarded as special patients with increased dental problems. The present paper consists of two parts. In the first part the types of hemophilia, ways of transmission, severity forms, and clinical characteristics are described. In the second part a protocol concerning the dental treatment of hemophilia patients is presented. There are four basic types of hemophilia: hemophilia A or classical hemophilia or Factor VIII deficiency, hemophilia B or Christmas disease, hemophilia C and von Willebrand's disease. Hemophilia is transmitted either as a sex-linked recessive or as an autosomal dominant trait, depending on the type of the disease. The severity of hemophilia depends on the amount of the coagulation factor present. According to this amount, there are four scales of severity. The clinical characteristics of the disease also depend on the amount of the factor present and vary, from occasional bleedings to serious and even life-threatening bleeding episodes. In the second part of the paper the special psychological and physiological problems of the hemophiliacs are discussed. In addition, there is reference to the hematologic coverage these patients need, as well as to the protection measures for the dental personnel against hepatitis and AIDS. The dental treatment plan at the office is presented in detail, including a discussion of the advantages and disadvantages of the treatment of hemophilia patients in the operating room under general anesthesia. PMID- 2978977 TI - [Hematoporphyrin derivative-laser induced changes in dehydrogenases in tumor bearing mice and patients with nasopharyngeal carcinoma]. PMID- 2978978 TI - [Visual evoked potentials in Down's syndrome]. PMID- 2978979 TI - [Desmoid tumors]. PMID- 2978980 TI - [Giant Ewing's sarcoma of the thoraco-abdominal wall]. PMID- 2978981 TI - [Oncogenes and oncogene products in urogenital cancer cells]. AB - Recent development in microbiology and genetic engineering has provided the identification and characterization of so-called 'oncogenes'. The concept of oncogenes has much stimulated intense interest in searching the cause of uncontrolled cell growth and factors responsible for formation of tumors. Because of the fact that oncogenes were first discovered in an established cell line derived from patient with bladder tumor, the association between oncogenes and genitourinary cancer has much attention. Variety of pathways of tumor development in bladder cancer can be divided in two major forms, low grade papillary tumor and high grade infiltrating tumor. Activation and a sequence of oncogenes may be relevant to the ultimate expression of these separate pathways. Concept of initiation and promotion may also be factored into these consideration. The application of these principles to the different pathways of tumor development such as in bladder, kidney and prostate cancers, supports the concept that oncogenes may be required to production of malignant tumors. The purpose of this paper is to review recent evidence that has enhanced our understanding of the genetic basis of cancer development in the genitourinary tract cancer. PMID- 2978982 TI - Investigations of the influence of mefenamic acid on histochemical reactions in the mucous duodenal membrane. PMID- 2978984 TI - A simple model of right ventricular hypertrophy. AB - Previous models of right ventricular hypertrophy (RVH) created by pulmonary artery (PA) banding in adult large animals have been associated with an unpredictable response of the right ventricle to the band and a high mortality due to the variable degree of acute stenosis. We studied the efficacy of PA banding in young pigs to produce RVH by progressive gradual stenosis during growth. Sixteen Yorkshire pigs at 6 weeks of age had nonconstricting 5-mm wide Dacron strips placed around the PA via a left minithoracotomy. The animals were returned for study in 2-3 months. There were no deaths during the growth period. Five sham-operated pigs acted as controls. Right ventricular free wall (RVFW) to total heart weight ratio was greater in the banded group (0.38 +/- 0.05 vs. 0.28 +/- 0.01, P less than .005) as was the RVFW to left ventricular free wall (LVFW) weight (1.09 +/- 0.25 versus 0.66 +/- 0.03, P less than .005). While the LVFW to total heart weight ratio decreased (0.36 +/- 0.04 vs 0.45 +/- 0.05, P less than .005), the septal ratio did not change (0.26 +/- 0.04 vs. 0.29 +/- 0.02, NS), indicating concomitant septal hypertrophy. This technique is simple, reliable, and reproducible in creating right ventricular and septal hypertrophy in the young pig with no mortality during maturation. PMID- 2978983 TI - Microvascular endothelial cell seeding of small-diameter Dacron vascular grafts. AB - One obstacle to the clinical implementation of endothelial cell seeding of vascular prostheses is the difficulty in derivation of large numbers of autologous endothelial cells from blood vessels of patients requiring vascular grafting. Capillary endothelial cells obtained from fat have been suggested as an abundant alternative to large-vessel endothelium for graft seeding. The object of this study was to evaluate the performance of 4-mm internal diameter (ID) Dacron Microvel grafts seeded with omentally derived microvascular endothelial cells. Six-cm lengths of the test grafts were implanted bilaterally into canine carotid arteries. One of each pair of grafts was seeded with endothelial cells (means = 8.4 x 10(6)) derived from collagenase digestion of autologous omental fat samples. The contralateral graft of each pair was nonseeded. At 5 weeks postoperatively, the grafts were harvested and evaluated. The mean patencies of both the seeded and nonseeded grafts were 89 percent. The mean thrombus-free surface area for seeded grafts was 95 +/- 11 percent. This value was significantly different statistically from the mean thrombus-free surface area of nonseeded grafts, which was 43 +/- 19 percent (P less than .05). Histologically, midgraft regions of seeded grafts were cellular, stained positive for collagen, and were characterized by inner capsules ranging in thickness between 35-94 microns. Luminal cells were identified as endothelial by peroxidase antiperoxidase staining techniques. Midgraft regions of nonseeded grafts demonstrated thrombus accumulation, limited cellularity, and inner capsules between 59-194 microns thick. Scanning electron microscopy of seeded grafts revealed smooth luminal surfaces with tight junctions between adjacent cells; surface cells were not present on midgraft regions of nonseeded grafts. In conclusion, endothelial cells derived from omental fat successfully surfaced on Dacron grafts and imparted characteristics to the graft that would predict long term graft success. PMID- 2978985 TI - A comparison of three techniques to quantify vascular ingrowth into Dacron velour. AB - Three methods of evaluation of vascular ingrowth into treated and untreated Dacron velour were performed to compare the methods. Vascular casting, direct measurement, and stereology were compared based on their ability to quantitate ingrowth, the type of information obtainable, and the time required for data acquisition. Results show that with the methods used in this study vascular casting was not able to produce data indicative of the actual vascular ingrowth. Direct measurement and stereological measurements of histological slides were able to give hard data for comparing ingrowth. However, stereological measurements were easier and less time-consuming to perform. In addition, stereological measurements were able to reveal additional parameters quickly. PMID- 2978986 TI - Some polymorphisms in the Salones. PMID- 2978987 TI - PGM3 polymorphism in human placenta samples from Bahia, Brazil. AB - PGM3 polymorphism was studied in 301 placenta samples from the Negroid population of Northeastern Brazil. The following gene frequencies were encountered: PGM3*1 = 0.545 and PGM3*2 = 0.455. They lie between European and African gene frequencies. The prevalent Black and White admixture of Northeastern Brazil populations fits well with the observed gene frequencies. Two samples exhibited variant electrophoretic patterns. PMID- 2978988 TI - PGM1 subtype distribution in Chinese population of the Guangzhou area. AB - PGM1 subtypes of 275 Chinese living in the Guangzhou area were determined using isoelectric focusing. The allele frequencies of PGM1 were 1+ = 0.6036, 1- = 0.1018, 2+ = 0.2200, and 2- = 0.0673. A rare variant, named PGM1*8, was found with a gene frequency of 0.0073. The phenotype distribution was consistent with Hardy-Weinberg equilibrium. PMID- 2978989 TI - Red cell enzyme markers in the Japanese of the Izena Island. AB - A genetic study was carried out on phenotype and gene frequencies of the genetic markers in four red cell enzymes, acid phosphatase (ACP1), esterase D (ESD), 6 phosphogluconate dehydrogenase (PGD) and phosphoglucomutase 1 (PGM1) among the Japanese population living on Izena Island of Okinawa Prefecture. The gene frequencies obtained were ACP1*A = 0.284, ACP1*B = 0.716, ESD*1 = 0.436, ESD*2 = 0.564, PGD*A = 0.947, PGD*C = 0.053, PGM1*1- = 0.112, PGM1*1+ = 0.602, PGM1*2- = 0.161, PGM1*2+ = 0.125. The population living on Izena Island is characterized by higher ACP1*A and PGM1*2 gene frequencies than those of Tokyo population. PMID- 2978990 TI - [The endocrine heart and gestational hypertension]. PMID- 2978991 TI - [The correlation of hysterosalpingography, Rubin tests and celiotomy with laparoscopic diagnoses in tubal permeability]. PMID- 2978992 TI - [Tubal microsurgery. The selection criterion and results]. PMID- 2978993 TI - [Laparoscopy in the Instituto Nacional de Perinatologia. An analysis of 738 cases]. PMID- 2978994 TI - Malignant melanoma in association with keratosis palmaris et plantaris (epidermolytic hyperkeratosis variant). PMID- 2978995 TI - Lentigo maligna with follicular mucinosis. PMID- 2978997 TI - Efficacy of the calcium antagonist isradipine in angina pectoris. AB - Thirty-six patients with chronic, stable angina pectoris were studied during 2 week treatment periods in which they received, in a randomized double-blind, crossover study, a new calcium entry blocking agent, isradipine, 7.5 mg three times daily or placebo. Antianginal efficacy was determined by treadmill exercise testing carried out 3 and 9 hours after drug administration on the final day of each treatment period. During placebo therapy, treadmill exercise time to the onset of angina (P1) and to the development of moderate angina (P2) was similar at 3 and 9 hours and similar to the placebo run-in period. During isradipine therapy, treadmill exercise time 3 hours after dosing was greater than with placebo therapy (P1 312 +/- 23.0 vs. 267 +/- 19.5 seconds, p less than 0.001; P2 410 +/- 20.2 vs. 355 +/- 18.8 seconds, p less than 0.002). Nine hours after drug administration, the results of exercise testing were similar to placebo. PMID- 2978996 TI - Atrial natriuretic factor: physiologic actions and implications in congestive heart failure. AB - Atrial natriuretic factor (ANF) represents a newly recognized hormone of cardiac origin. This peptide is synthesized by the myocardial cells of both atria and released by atrial stretch. The hormone promotes sodium and water excretion by the kidney, inhibits the renin-angiotensin-aldosterone system, and reduces systemic arterial pressure. Specific receptors for ANF are present in the kidney, adrenal glands, vascular smooth muscle, platelets and central nervous system. Congestive heart failure is characterized by increased circulating levels of ANF; however, there appears to be an attenuation in the renal response to the hormone. Recent investigations have reported the effect of systemic administration of synthetic ANF to normal individuals and those with congestive heart failure. The hormone may promote a significant natriuresis and diuresis in addition to reducing arterial pressure and inhibiting renin and aldosterone secretion. Substantial questions remain as to the full physiologic significance and therapeutic potential of this hormone. PMID- 2978998 TI - [Oncological laparoscopy]. PMID- 2978999 TI - Donor-specific transfusion: a report of the Japanese Renal Transplant Registry. AB - 1. The effect of donor-specific blood transfusions (DST) on renal graft survival of HLA one haplotype-mismatched living-related transplant patients was studied, and 1,292 transplant patients reported in the Japanese Renal Transplant Registry were analyzed. In the CsA groups, the graft survival rate at 4 years of the DST, DST plus unspecified blood transfusion (UTF), UTF, and no blood transfusion (TF = 0) groups, were 93.5%, 91.6%, 76.2%, and 62.7%, respectively, whereas, in the conventional groups, the graft survival rate at 4 years of the DST, DST plus UTF, UTF, and TF = 0 groups were 73.3%, 87.2%, 73.2%, and 69.0%, respectively. 2. The superior DST effect was prominent in the transplants on CsA, but not in those on conventional immunosuppression. The DST patients receiving UTF, however, showed a superior DST effect. 3. DST patients treated with conventional immunosuppression and the CsA patients receiving UTF showed an equal graft survival rate. 4. Since the CsA therapy was able to achieve a higher graft survival rate in a high MLR combination (S.I. greater than 10), it is likely that the DST patients treated with CsA showed an excellent graft survival rate. 5. The transplant patients without blood transfusion in the CsA and conventional groups achieved a moderate graft survival rate. PMID- 2979000 TI - [Second-look laparoscopy in gynecology]. PMID- 2979001 TI - Expression of the gene for the atrial natriuretic peptide in cardiac myocytes in vitro. AB - Primary cultures of neonatal cardiocytes express the gene for atrial natriuretic peptide (ANP). In general the levels of expression follow the rank order: atrial greater than ventricular much greater than nonmyocardial cells. Following the initial dispersion cardiocytes require 48 to 72 hours before ANP secretion and ANP mRNA accumulation approach a new steady-state level. In situ hybridization analysis indicates that ANP gene expression is concentrated in a subpopulation of cardiocytes in both the atrial and the ventricular cell cultures. These findings suggest that these primary cultures may be of value in defining the factors governing the expression of the ANP gene in the cardiac cell. PMID- 2979002 TI - Recruitment of the left ventricle for production of atrial natriuretic factor in Dahl salt-hypertension sensitive rats. AB - Dahl salt-sensitive (S) rats on normal salt intake develop a chronic form of hypertension with aging whereas Dahl salt-resistant (R) rats do not. In old hypertensive S rats the left ventricle is recruited for the production of ANF as evidenced by a seven- to ten-fold increase in atrial natriuretic factor (ANF) mRNA and increased ANF in ventricular tissue. Plasma ANF is also markedly increased in hypertensive S rats but there is no direct proof that ventricular production of ANF contributes to high plasma ANF. Atrial ANF is higher in S than R rats but this strain difference is not associated with strain differences in ANF mRNA in atria and atrial ANF mRNA does not increase dramatically in hypertensive S rats as does ventricular mRNA. Recruitment of the ventricle to produce ANF is clearly a secondary consequence of hypertension, but its functional significance is not established. PMID- 2979003 TI - A pharmacologic study of the sympathetic eyelid tarsal muscles. AB - The contractile state of the sympathetically innervated superior and inferior tarsal muscles, also known as Mueller's muscles, was defined in 10 young adults by stimulating and then paralyzing these muscles pharmacologically in a randomized, double-masked study, using one eye for drug and the other for control. The drugs used topically were phenylephrine hydrochloride 2.5 and 10% for muscle stimulation, and guanethidine 5% and thymoxamine hydrochloride 0.5% for muscle inhibition. A model is described for accurately examining and recording changes in the vertical palpebral fissure simultaneously bilaterally using magnified images from two high-resolution, high-sensitivity video cameras. The superior tarsal muscle averaged 50% of its total possible contraction when it was in its tonic state, and the inferior tarsal muscle averaged 67%. The drug instillations defined the mean maximum total excursion of the superior tarsal muscle as 3.0 mm, with 1.5 mm of upward displacement and 1.5 mm of downward displacement from its tonic position. The inferior tarsal muscle had a mean maximum total excursion of 0.3 mm, with 0.1 mm downward displacement and 0.2 mm upward displacement from its tonic position. PMID- 2979004 TI - Interleukin-2 effect on the inhibition of mixed lymphocyte reaction induced by cyclosporin A. PMID- 2979005 TI - Photoaffinity labeling of pyruvate kinase from rabbit muscle. AB - Several studies have been performed on the structure of muscle pyruvate kinase. X ray diffraction has provided a three-dimensional picture of the active site, and chemical modification studies have revealed essential amino acid residues for substrate binding or catalysis. We have shown that 8-azido-ADP (N3 ADP) behaves as a photoaffinity label for the enzyme. This reagent upon irradiation produces inactivation of the enzyme, and the activity loss is protected by nucleotides. The partially modified enzyme shows the same Km for ADP as the native one suggesting an "all or none" inactivation effect. The incorporation of 1 mole of 14C-N3 ADP per subunit correlates with complete inactivation. A radioactive peptide was isolated from the enzyme labeled with 14C-N3 ADP. The partial sequence of this peptide showed that it corresponds to the same peptide isolated from rabbit muscle pyruvate kinase labeled with dialdehyde-ADP and with trinitrobenzenesulfonate. This peptide is identical to a region in the cat and chicken muscle enzymes, and also a high degree of homology is found in a region of the rat liver and yeast enzymes. These studies show that N3 ADP binds to the same site as dialdehyde-ADP in rabbit muscle pyruvate kinase, and this site seems to be the nucleotide binding site. PMID- 2979006 TI - Proteolysis of mitochondrial-coded and nuclear-coded proteins found in yeast mitochondria. AB - The rate of degradation of radioactive labeled mitochondrial proteins synthesized both in vitro and in vivo by isolated yeast mitochondria and growing yeast cells respectively, has been studied. It was found that the in vitro-synthesized mitochondrial proteins are rapidly degraded by an energy-dependent proteolytic system. Under the same experimental conditions the in vivo-synthesized mitochondrial proteins are slowly degraded to a limited extent by a protease which is slightly inhibited by ATP. During this period, the mitochondria are coupled and metabolically active. It is proposed that mitochondria possess an energy-dependent proteolytic system that recognizes as substrates either "abnormal" proteins or unassembled protein subunits encoded in the mitochondrial genome. An apparently different system, which is independent of energy, seems to be responsible for the slow and limited degradation of "normal" mitochondrial proteins. PMID- 2979007 TI - [Effect of experimentally-induced stomach ulcers in rats on the histochemical reactions in the duodenal mucosa]. PMID- 2979008 TI - [Cytomorphological studies of the effect of Feldene on gastric mucosa of rats]. PMID- 2979009 TI - [Vibration-induced angiopathy: significance of urinary beta-thromboglobulin increase after cold test]. AB - Twenty subjects working with tools generating 100-400 Hz vibrations were submitted to volumetric pulse-plethysmography with cold Test (C.T.) and to the measurement of BTGU, a thromboxane urinary catabolite. After C.T. an increase of BTGU was observed, which attained statistical significance in the subgroup with the most evident plethysmographic modifications. The meaning of these findings is discussed, also in the light their apparent contrast with some literature data. The AA believe that the prostaglandin balance can actually play a role in the pathogenesis of C.T.-induces vasomotor changes, if the tools used generate a sufficiently high number of Hz and if the workers are still rather young. This last condition corresponds to a greater intactness of endothelia and thus to the ability to modulate the vasomotor response ("hunting reaction"). This would lead to the activation of more complex pathogenetic mechanism (also biohumoral ones) among which is thromboxane release. PMID- 2979010 TI - The surface phenotype of activated T lymphocytes. AB - The surface phenotype of T cells reflects both their relative maturity and their activation state. To determine the pattern of surface markers characteristic of activated T cells, purified mature T cells were stimulated in vitro for periods of 0.5-5 days with Concanavalin-A (Con-A) or phorbol myristic acetate (PMA) and ionomycin, fluorescein-labelled with monoclonal antibodies, then analysed by flow cytometry. The level of expression of the function-associated antigens CD4 (L3T4) and CD8 (Ly-2) decreased transiently early after activation with PMA/ionomycin, but not after stimulation with Con-A. Both stimuli caused a small drop in the level of CD3 and the T cell antigen receptor (TCR). At no time was CD3, CD4 or CD8 completely lost from the surface. Following activation Pgp-1, the interleukin 2 (IL-2) receptor (as detected by the monoclonal antibody 7D4) and the peanut agglutinin (PNA) receptor were gained by a proportion of cells, MEL-14 was lost by a proportion of cells, and no change was observed in the expression of heat stable antigen. Thy-1 or Ly-1. From the present data no evidence has been found for the generation of the 'immature', CD4- CD8- phenotype found in the thymus by activation of mature T cells. During T cell development, however, changes in expression of Pgp-1, MEL-14, the IL-2 receptor and the PNA receptor may be associated with activation, rather than differentiation per se. PMID- 2979011 TI - [Inhibition by ethanol and acetaldehyde the plasmin activity and plasminogen activation induced by urokinase and streptokinase]. AB - Ethanol and in a greater degree acetaldehyde inhibit activation of plasminogen evoked by urokinase and streptokinase. Ethanol does not inhibit the plasmin caseinolytic and fibrinolytic activities though the former inhibits amidolytic activity of the enzyme but only insignificantly. Acetaldehyde inhibits the plasmin activity towards casein, fibrin and H-D-Val-Leu-Lys-pNA. PMID- 2979012 TI - Crouzon syndrome. A review of literature and case report. AB - A sporadic case of Crouzon Syndrome without mental retardation is described. The patient, an 11 year old boy with grossly carious teeth and in severe pain was referred for dental treatment. A review of literature on Crouzon Syndrome is presented. PMID- 2979013 TI - STEM analysis of the dynein ATPase. PMID- 2979014 TI - The immunoregulatory disturbance in autoimmune thyroid disease. AB - There is now considerable evidence for a genetically induced antigen-specific defect in suppressor T lymphocytes as the basis for AITD, derived from several laboratories and via different types of experimental techniques. In addition, there is now evidence for additive effects on reducing generalized suppressor T lymphocyte numbers and/or function by environmental factors as well as hyperthyroidism itself, and these effects would be superimposed upon the organ specific defect. Such effects on generalized suppressor T lymphocyte numbers may act as precipitating and self-perpetuating factors, in Graves' disease at least. Presentation of the antigen by the thyroid cell via HLA-DR expression on its membrane does occur as a result of interferon gamma production by T lymphocytes. This in turn appears to be secondary to the initial specific immune assault and is not a primary inductive step. However, it may be important as an amplifying intermediate factor but cannot perpetuate the process in the absence of the underlying immune disorder. There is, however, no evidence for an underlying antigenic abnormality or stimulus in human autoimmune thyroid disease, and the initiating event would appear to be due to perturbation of the generalized immune system superimposed on the organ-specific abnormality. Variations in the serological and clinical expression of AITD would appear to depend on the severity of the original organ-specific disturbance in suppressor T lymphocyte function, plus the added factor of environmental influences playing upon generalized suppressor T lymphocyte function and numbers. Remissions in Graves' disease brought about by antithyroid drugs may well be via their effect on modulating the thyroid cell activity; this will then reduce thyrocyte-immunocyte signalling, allowing remission to occur in those patients with a partial organ specific defect in suppressor T lymphocytes. PMID- 2979015 TI - Commentaries on "The immunoregulatory disturbance in autoimmune thyroid disease". PMID- 2979017 TI - The orbital surgeon. AB - While the number of orbital surgeons is limited, it is hoped these can be recognized and patients referred to them by ophthalmologists not interested or trained in that specialty. Let the orbital surgeon determine whether he can handle the problem in 1 to 2 days, or whether a neurosurgeon should do the procedure or make it a joint effort. It may well involve other specialty team effort approaches. It is essential to have an understanding of x-rays, CT, angiography, and MRI techniques and films. Sit with these specialists to learn more and help to avoid negative, misdiagnosis reports in the interest of the patient. Use judgement in helping the patient decide on ophthalmic or the more extensive neurosurgical approach after careful study and what is in their best interest. The team approach is used in well established medical centers with the ophthalmologist and neurosurgeon (or other specialist) working together in the best interest of the patient. This is more interesting and keeps the ophthalmologist in the mainstream of medicine. PMID- 2979016 TI - Atypical cases of sinus histiocytosis (Rosai-Dorfman disease) with ophthalmological manifestations. AB - Sinus histiocytosis with massive lymphadenopathy is a non-neoplastic disease of unknown cause observed mainly in children with markedly enlarged cervical or other lymph nodes. We have reported two very atypical cases with extranodal manifestations that required ophthalmological consultation. Neither patient had significant lymphadenopathy. One patient, who had tumors of all four eyelids as well as lesions of the nose and larynx, was 68 years old when first examined, and 70 at the time of his last surgical procedure. The other patient, a 13-month-old child, had a unique corneoscleral lesion as his only clinical manifestation. The excised tissues obtained from the eyelids and larynx in case 1 and from the corneoscleral lesion in case 2 revealed characteristic histopathologic features of sinus histiocytosis (Rosai-Dorfman disease), including strongly positive immunoperoxidase staining for S-100 protein in the cytoplasm of most of the histiocytes. PMID- 2979018 TI - A re-evaluation of corneal development. AB - The corneal epithelium is initially a continuation of the surface ectoderm, but later on in development appears to arise from the superficial cells of the corneal stroma. The corneal epithelium varies in thickness depending on the status of the lids, viz either fused or open. When the lids are fused, the epithelium is only 2 to 3 layers in depth. When the lids are separated, a basement membrane is distinguishable and the epithelium is 4 to 5 layers in depth. Bowman's membrane develops from processes of the superficial mesenchymal cells of the stroma that become thickened and are arranged in the long axis of the corneal surface. The corneal stroma develops from in situ mesenchymal tissue and does not migrate from the limbal mesenchymal tissue. The attenuation of the central cornea, early in development, is due to the impingement of the lens against the developing cornea. The central constriction of the cornea has led previous observers to believe that the stroma migrates from the peripheral limbal area towards the center of the cornea. Descemet's membrane arises from processes of the deep mesenchymal cells of the corneal stroma. These processes thicken and become arranged in the long axis of the posterior surface of the cornea. The membrane is initially cellular, and well-defined septae are noted between the cells. With maturity, Descemet's membrane becomes a homogeneous structure. The endothelium is derived from the mesenchymal cells of the posterior stroma. These cells are initially cuboidal but then become flattened. During the early development of the cornea, the tunica vasculosa lentis may play an important role. It is suggested that the anterior chamber is maintained early on by a transudate from the vessels of the tunica vasculosa lentis. The vessels of the tunica vasculosa lentis are compressed by the vanguard of the optic cup against the equator of the lens. With regression of this vascular system, there is a simultaneous development of aqueous humor. The hydrodynamic force of aqueous production assists corneal molding by a vis a tergo affect. Pari passu with this hydrodynamic force, the corneal stromal fibers increase in length and width. The structures involved in aqueous humor production, viz the ciliary epithelium, and the aqueous humor drainage, viz the filtration angle, trabecular meshwork and aqueous veins, develop contemporaneously as the tunica vasculosa lentis regresses. The limiting and fixed stabilizing site is at the limbus, the site of insertion of the rectus muscles. At these sites a dimpling occurs as the cornea is enlarging. PMID- 2979019 TI - Recovery of visual acuity following the repair of pseudophakic retinal detachment. AB - Postoperative visual acuities were evaluated in a series of 100 pseudophakic eyes in which rhegmatogenous retinal detachments involved the macula and in which reattachment surgery was successful. Preoperative visual acuity and duration of macular detachment were directly and indirectly related to visual outcome, respectively. Eyes in which extracapsular surgery had been followed by posterior chamber lens implantation had significantly better postoperative visual acuities than cases in which older iris-fixation IOLs were placed following intracapsular procedures. PMID- 2979020 TI - Retinal detachment: is reparative surgery always mandatory? AB - The author reports 16 cases of rhegmatogenous retinal detachment in which, for a variety of reasons, immediate reparative surgery was not carried out. To date, one-half of these cases have not been operated, and four have been followed for 6 years or longer. No case suffered visual loss because of the delay in surgery, nor has chronic inflammation, glaucoma, or rubeosis been a problem. The clinical characteristics of this group of cases is defined. PMID- 2979021 TI - The pathology of anterior (peripheral) proliferative vitreoretinopathy. PMID- 2979022 TI - Acute exudative polymorphous vitelliform maculopathy. PMID- 2979023 TI - Progressive visual loss in adults with retinopathy of prematurity (ROP). PMID- 2979024 TI - Distant effect of peripheral branch vein occlusion on the macula. PMID- 2979025 TI - Retinal and choroidal infarction from Aspergillus: clinical diagnosis and clinicopathologic correlations. PMID- 2979026 TI - Esotropia. PMID- 2979027 TI - Glaucomatocyclitic crises and systemic disease: peptic ulcer, other gastrointestinal disorders, allergy and stress. PMID- 2979028 TI - Fine needle aspiration biopsy of suspected metastatic cancers to the posterior uvea. AB - This thesis presents the author's experience with diagnostic intraocular fine needle aspiration biopsy in 18 patients with a suspected metastatic choroidal or ciliary body tumor. The author has reviewed the literature on biopsy of intraocular tumors and has specified what he believes to be valid indications for diagnostic biopsy of posterior uveal tumors. He has evaluated the accuracy, limitations, and complications of diagnostic fine needle aspiration biopsy in this series and others, and he has suggested methods for improving the recovery of sufficient cells for cytologic diagnosis and lessening the risks of tumor cell seeding during the biopsy. The author has concluded that fine needle aspiration biopsy appears to be a relatively safe, generally reliable means of establishing the pathologic diagnosis of a choroidal or ciliary body tumor in highly selected patients suspected of having metastatic cancer. In spite of its apparent safety and reliability, however, the author has cautioned against the routine use of fine needle aspiration biopsy in patients with posterior uveal tumors since its long-term safety has not been established. The author has suggested that diagnostic fine needle aspiration biopsy of posterior uveal tumors be performed only in medical centers where there can be input from and cooperation among ophthalmologists, ophthalmic pathologists, and cytopathologists who are experienced in the diagnosis of intraocular malignancies. PMID- 2979029 TI - Reconstruction of the lids of a child with microblepharon and multiple congenital anomalies. AB - The initial stages in the rehabilitation of a male child with severe microblepharon, corneal opacities, bilateral facial clefts, bilateral complete cleft lip and palate, and unilateral syndactyly are described. Review of the literature suggests that severe microblepharon is associated with other craniofacial anomalies, and often the child is stillborn or retarded. Surviving children have been abandoned because of their appearance. The child described in this case appears to be unique because his intelligence is normal, and, to our knowledge, this is the first reported case of penetrating keratoplasty after reconstruction of functional eyelids. The principal problems after corneal grafting appear to have been chronic partial exposure due to inadequate lid length and a poor Bell's reflex and the persistence of a rim of vascularized fibrous tissue around the corneal graft. Future reconstructive surgery is outlined. PMID- 2979031 TI - Iris pigmentation and pigmented lesions: an ultrastructural study. PMID- 2979030 TI - DNA cell cycle studies in uveal melanoma. AB - We analyzed the cell cycling status of a group of irradiated and nonirradiated uveal melanomas using BrdUrd techniques. These data demonstrate that melanomas are relatively slow-growing tumors with a few cells actively cycling at a given time. Radiation has a profound effect on the number of cycling cells (P less than .0001). After treatment with either 20 Gy of pre-enucleation photon or 60 Gy or more of helium ion irradiation, virtually no cells are detected in the synthesis phase of the DNA cell cycle. It is unclear whether the absence of cycling cells after 20 Gy of photon irradiation is permanent or represents a transient cell cycle block, since these tumors were studied within 48 hours after irradiation. In contrast, all melanomas treated with helium ion had been irradiated several months prior to enucleation (mean, 2 years). In the latter group of tumors, the length of time between treatment and cell cycle analysis suggests that these cells had lost their reproductive integrity. These data were substantiated by tissue culture studies. Growth of tumor explants was significantly less (P less than .007) in irradiated than in nonirradiated melanomas. The optimum technology used for measurement of cell cycle status remains to be determined. Measurement of BrdUrd uptake using immunofluorescent microscopy on either standard sections or fine-needle biopsies can be performed. In general, flow cytometric analysis yields similar results. It is difficult with the latter technique to be certain that nontumor cells are not artifactitiously counted in the cell cycle studies. The incorporation of BrdUrd cell cycle analysis with fine-needle biopsy may be useful in the clinical management of irradiated melanomas that have questionable growth after treatment. In a few cases studied, results appeared to correlate with tumor growth activity. PMID- 2979032 TI - Upper eyelid retraction in Graves' ophthalmopathy: a new surgical technique and a study of the abnormal levator muscle. AB - A new surgical procedure, the proximal levator technique, achieves recession of the retracted upper eyelid in Graves' ophthalmopathy by sectioning the levator muscle proximal to Whitnall's ligament and fixing eyelid position with sutures that permit postoperative adjustment. This technique deserves further study. Enlargement of the proximal levator muscle in Graves' eye disease is shown on orbital CT scans and is found at surgery when the proximal levator technique is employed. Histologic and morphometric studies demonstrate increased levator muscle fiber size as well as increased extracellular volume. These findings suggest that levator muscle hypertrophy is important in the pathogenesis of upper eyelid retraction in Graves' ophthalmopathy. PMID- 2979034 TI - Strategies for the initial management of acute preseptal and orbital cellulitis. PMID- 2979033 TI - The ciliary vasculature and its perturbation with drugs and surgery. PMID- 2979035 TI - The decreased incidence of retinal detachment after cataract surgery. PMID- 2979036 TI - Epidemiology and natural history of ocular herpes simplex virus infection in Rochester, Minnesota, 1950-1982. PMID- 2979037 TI - Digestive cancers and nutrition in Belgium. PMID- 2979038 TI - Gastric and duodenal ulcer in the same patient. AB - No extensive endoscopic studies have been performed on the prevalence and the clinical outcome of association of gastric ulcer (GU) and duodenal ulcer (DU). The present investigation, partially retrospective and partially prospective, takes into account 715 patients with active ulcer demonstrated by endoscopy, followed-up for a mean period of 3.8-years; 23 of them (3.2%) were found to have synchronous or asynchronous gastric and duodenal ulcers. The following characteristics were investigated: age of onset of both diseases, ulcer family history, cigarette and alcohol consumption, nonsteroidal anti-inflammatory drugs abuse, serum pepsinogen group I, ABO and Lewis blood groups, healing and relapse rate under H2-blocker treatment. The first diagnosis (by either X-Ray or endoscopy) was DU in 500 subjects (70%), GU in 210 (29.3%) and synchronous gastric and duodenal ulcers in 5 (0.7%). After a median period of 10 years, 2.8% of DU patients developed a GU; after 2-12 yrs 1.9% of GU patients developed a DU. The clinical and biochemical findings of our GU/DU patients suggest that the two ulcers are related by chance. IN CONCLUSION: asynchronous GU/DU patients do not seem to have a distinct disease in the large spectrum of ulcer disease. Larger studies must be planned on synchronous GU/DU with the aim of assessing whether or not it represents a particular type of ulcer disease. PMID- 2979040 TI - [Inverse polypoid hamartoma of the rectum with colonic extension of the lesions and deep cystic colitis with rectal involvement. Apropos of an anatomo-clinical case report]. AB - The authors report the case of a 38 year old man, presenting with an inverted polypoid hamartoma (IPH) of the rectum, associated with similar colonic localizations and they show the resemblance of these lesions to the colitis cystica profunda (CCP) with involvement of the entire large bowel and eventually the rectum. In fact, moreover, the symptoms, the digital rectal examination and the endoscopic, radiological and pathological data are similar but, in the reported cases, the IPH always included at least one rectal localization whereas the CCP might not show rectal lesion. Advanced lesions of rectal IPH may be confused, by endoscopy and pathology with adenomas or adenocarcinomas. Consequently it is essential to perform large and deep biopsies for histological diagnosis. The IPH and the CCP are benign diseases but "recurrence" may be observed in case of incomplete removal. The removal has to be adequate but not mutilating, either by surgery or by laser endoscopic photoablation. The choice of the procedure will depend upon the size of the lesion. PMID- 2979039 TI - Campylobacter pylori-associated gastritis: attempts to eradicate the bacteria by various antibiotics and anti-ulcer regimens. AB - The efficacy of various antimicrobial and anti-ulcer agents on the eradication of Campylobacter pylori in patients with antral gastritis or duodenal ulcers was investigated by several open studies or double-blind, placebo-controlled protocols. Among the anti-ulcer agents, ranitidine, cimetidine or sucraflate had no effect on C. pylori. Colloidal bismuth subcitrate achieved clearance of C. pylori in 40% of treated patients at the end of therapy but a high relapse rate (14/16 patients) was observed after a 6-month follow-up period. The antibacterial agents doxycycline, minocycline, ofloxacin, clindamycin, paromomycin and nifuroxazide failed to eradicate C. pylori in most patients. By contrast, short term elimination of C. pylori could be achieved in more than 90% of patients treated with amoxycillin. However, relapse occurred as a rule in all amoxycillin treated patients within one month after therapy. Overall, we observed no correlation between the in-vitro activity of the different antibacterial agents and their in vivo efficacy. Development of resistance during therapy does not seem to account for this discrepancy since it occurred only with ofloxacin. On the basis of these results, we conclude that long term eradication of C. pylori from the gastric antrum cannot be achieved after monotherapy either with antibiotics or with bismuth salts. PMID- 2979041 TI - [Stereomorphological relationship between colonic adenomas and peri-adenomatous mucosa in scanning electron microscopy. Histogenetic correlations]. AB - Using scanning electron microscopy (SEM), then light microscopy, the authors have studied 25 colonic adenomas, 5 specimens of peri-adenomatous mucosa and 9 specimens of normal colonic mucosa. Comparison between SEM features and histological characteristics has shown that each of the three histological entities had an specific surface architecture. Comparative analysis of the different aspects observed at SEM has shown a close architectural relationship between the adenomas and the adjacent mucosa, and suggests the hypothesis that adenomatous polyps are formed by invagination of the surface epithelium which has a well-known capacity of proliferation. PMID- 2979042 TI - [Risk of recurrence of colorectal polyps following endoscopic resection]. AB - From 1976 to 1985, 1063 patients (614 men, 449 women, mean age: 63 years) underwent the excision of 1887 adenomatous or villous colonic or rectal polyps. Regular controls by barium enema or total colonoscopy have been proposed to these patients. A first control, performed in 715 patients (67%) after a mean of 27 months, revealed the presence of polyps in 162 of them (23%). A second control performed in 331 of the 535 controllable patients (61%), was positive (presence of polyps) in 82 of them (24%). During the follow-up period, 16 cancerous lesions were observed. Comparing the initial status of the patients with the results of the different controls, 3 risk factors for developing a new colonic or rectal polyp emerged: 1) male sex, 2) the presence of multiple lesions at the initial examination, 3) a recurrence at a previous control. PMID- 2979043 TI - [Polyps with an invasive degenerated focus]. AB - Advances in technology of flexible endoscopes have greatly changed the management of patients with adenomatous polyps of the colon and rectum. Some controversy still exists concerning the best treatment for invasive polyps. For some authors, invasive polyps need radical operation, while others think that unless cancer goes beyond the bounds of a removed polyp, endoscopic resection is an adequate procedure. We designed a study of 65 patients presenting an invasive carcinoma arising in adenomatous polyps and who underwent a colorectal resection thereafter, in order to determine which endoscopic and histological features correlated best with a curative treatment by polypectomy. When the group of "non curative polypectomies", (carcinoma in the surgical specimen: 34 patients) was compared to the group of "curative polypectomies" (carcinoma in the surgical specimen: 31 patients), there was no significant difference in the number of pedunculated or sessile polyps but a polyp's size exceeding 30 mm was significantly more frequent in the group of "non-curative polypectomies" (P less than 0.005) as well as a tubulo-villous or villous histological type (P less than 0.001) and presence of vascular neoplastic invasion (P less than 0.01). In conclusion, a surgical resection after endoscopic polypectomy of a polyp containing an invasive carcinoma is necessary for a polyp's size exceeding 30 mm, for a villous or tubulo-villous type and in the presence of vascular neoplastic invasion in the pathological analysis of the removed polyp. PMID- 2979044 TI - [Villous tumors of the rectum. Endoscopic treatment using monopolar coagulation]. AB - Many methods are used to treat villous adenomas of the rectum. The best choice between these methods is not well established. Ninety-six patients (47 men, 49 women) treated with endoscopic monopolar coagulation were studied. The mean age was 66 years (range 37-84). The lesions were located in the lower third of the rectum in 27 cases, in the middle third in 35 cases and in the upper third in 34 cases. The circumferential extent of the lesions was less than 1/3 in 70 cases (C1), greater than 1/3-less than 2/3 in 23 (C2) and greater than 2/3 in 3 cases (C3). The follow-up of 2 patients (1 C1 and 1 C2) was not sufficient. Healing was not achieved in 8 of the 94 evaluated patients. Eighty-six patients were totally healed: 68 C1, 16 C2 and 2 C3 with a follow-up of 2 to 132 months (median: 36 months). The treatment was achieved after a mean of 2.1 sessions per patient (range 1-13). CONCLUSIONS: 1) Endoscopic monopolar coagulation of villous adenoma of the rectum is a simple and efficient treatment. 2) This method is complementary to surgery for extensive lesions (C3). PMID- 2979045 TI - [Villous tumors of the rectum. Endoscopic treatment using photocoagulation]. AB - We report the results of endoscopic photocoagulation in the treatment of villous adenomas of the rectum in 49 consecutive patients (26 males, and 23 females, mean age: 74 years). Twenty-six patients were treated with a high-power Nd-Yag laser (80 W/sec). After a mean of 3.4 laser sessions, all the small-sized adenomas (C1) and 88% of the intermediate-sized adenomas (C2) were healed. No extensive villous adenoma (C3) could be eradicated. Complications occurred in 5 patients. Recurrence was observed in 3 C2 patients who are still on maintenance laser therapy. Twenty-three patients were treated with another Nd-Yag laser (maximal power output: 45 W/sec). After a mean of 6.4 laser sessions, 85% of the C1 patients were healed, while eradication of villous tissues was obtained in 60% of C2 and C3 patients. No complications were noted. A recurrence was observed in three C1 patients with secondary healing after reinstitution of laser therapy in 2 cases. Moreover, two C3 patients also relapsed and laser eradication was again obtained in 1 patient. In conclusion, these results confirm the efficacy of laser therapy in small- and intermediate-sized villous adenomas of the rectum. The number of laser sessions required for eradicating a villous tumour was lower using a high-power Nd: Yag laser, but the risk of complications was increased. PMID- 2979047 TI - Members. American Ophthalmological Society. PMID- 2979046 TI - [Surgical approach to villous tumors of the rectum]. AB - The authors report the results of the surgical treatment of 39 villous adenomas of the rectum operated on during a 10-year period. Twenty-nine tumours were treated by a local excision, and 10 by a subtotal rectal resection. The choice for the procedure was mainly determined by the level of the lower tumour margin. Thirty-four operations were performed with a curative goal, and the 5 others for debulking. There was no operative mortality, and the morbidity was very low. Four invasive carcinomas were detected in the operative specimens and the four patients were withdrawn from the follow-up. In the "curative group", local recurrence rate was 21% after a mean follow-up period of 46 months. The tumour recurrences were small, and 2/3 among them were easily treated by reoperation (1 case) or further coagulations. With some complementary coagulations, surgery yielded a full control of 100% of the C1, 83% of the C2 and 66% of the C3 lesions. Full tumour clearance was achieved in two patients of the debulking group, with some postoperative coagulations. Our series and the literature confirm the efficiency of the surgical approach, which, moreover, allows the best histological assessment in one single session. PMID- 2979048 TI - Clinical manifestations of ectopia lentis et pupillae in 16 patients. AB - The largest known series of patients with ELeP was clinically evaluated. Sixteen patients from eight families were characterized with respect to ocular anomalies. Variability between eyes of the same patient and among different patients was marked. In addition to the well-known but inconstant displacement of the lens and pupil, other characteristics of this protean syndrome included severe axial myopia (and associated fundus abnormalities), poor vision, retinal detachment, enlarged corneal diameters, cataract, abnormal iris transillumination, poor pupillary dilatation, persistent pupillary membranes, iridohyaloid adhesions, and prominent iris processes in the anterior chamber angle. PMID- 2979050 TI - Effectiveness of apraclonidine in preventing the rise in intraocular pressure after neodymium:YAG posterior capsulotomy. AB - Apraclonidine (para-aminoclonidine) is an alpha agonist that was studied for its effect on the IOP rise following YPC. In a prospective multicentered double masked study 63 eyes were pretreated with one drop of either 1% apraclonidine or placebo 1 hour prior to performing YAG and again after the laser treatment. The greatest IOP rise in the placebo-treated eyes occurred in the third hour after YPC when the mean IOP rose from a baseline pressure of 16.4 +/- 3.7 mm Hg to 20.8 +/- 6.8 mm Hg (P less than .01). In apraclonidine-treated eyes the IOP fell from a mean of 15.6 +/- 3.8 mm Hg to 12.8 +/- 6.0 mm Hg 3 hours postoperatively (P less than .001). There were five times as many eyes that had a pressure rise greater than 10 mm Hg in the placebo-treated group compared to those treated with apraclonidine. Apraclonidine proved to be highly effective in preventing the rise in IOP following YPC. PMID- 2979049 TI - Human excimer laser corneal surgery: preliminary report. AB - The first human trial utilizing the argon fluoride excimer laser at 193 nm to produce a superficial keratectomy in ten human eyes has been described with the histopathological evaluation of four eyes and the longer gross appearance of six eyes at intervals extending to 10 months post-excimer laser treatment. The process of laser superficial keratectomy has proved to be one of the promising areas of surgical intervention for reconstructive or refractive keratoplasty in the future. Intensive investigations need to be undertaken on the corneal wound healing process following laser ablation as well as the nature, and long-term stability of the corneal excisions or induced refractive corrections. It is essential that the optimal laser parameters be established for the various refractive corrections and other corneal surgical techniques, and that pathophysiologic and histopathologic changes that have been induced by the excimer laser-corneal tissue interaction in animals and humans be critically and extensively analyzed. PMID- 2979051 TI - [Dietary lipids and experimental colonic carcinogenesis. Critical review of the literature]. AB - Colon cancer is one of the most common tumours observed in the western population for which the relationship between epidemiological and laboratory findings and an overall assessment of the influence of diet on carcinogenesis is not straightforward. The aim of this review is to evaluate critically the experimental data which suggest a positive modulating effect of dietary lipids. Although a great number of data are available on the relation between the nature of fat in diet and colon cancer, no clear conclusions can be drawn from their analysis. The lack of consistent findings is due to the discrepancy of the results which may have arisen from methodological differences between the studies. Concerning the influence of the amount of dietary fat on colon cancer, it has been demonstrated that a high fat diet could increase the incidence and the number of colonic tumours in rats. Nevertheless, this positive modulating effect could be seen provided the fat amount in the diet was at least 20% (w/w). This observation is of first importance for colon cancer prevention and must be confirmed for fat of various nature. Systematic dose-effect studies are necessary. The question whether the effect of dietary fat on carcinogenesis is due to the specific action of fat or to an associated caloric effect has been raised several times. Although the previous studies concerned the role of caloric intake versus fat intake in carcinogenesis, the observed effects might have been due to a change in body weight or to changes in other dietary components such as non nutritive fibre, protein and micronutrients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979052 TI - [Histology of colonic polyps]. AB - The authors present the modern classification of colorectal polyps. They discuss the dysplastic and preneoplastic changes in the light of new data on predisposing factors. Although new techniques are developing such as immunocytochemistry, DNA cytophotometry, DNA recombination, it is clear that presently, the basic morphological data, microscopical as well as histological, retain an irreplaceable diagnostic value. From a practical point of view, the authors recall the importance of an adequate orientation of the polyps before their histological examination in order to obtain all the necessary informations permitting an accurate diagnosis: type of the polyp, possible areas of cancer formation and their depth of penetration, histology of the surrounding mucosa and tissues. PMID- 2979053 TI - [[Evaluation of various methods of studying fecal occult blood]. AB - Occult blood testing for detection of asymptomatic colorectal cancer is an old concept. The authors review the literature about the screening value of current tests. Among the chemical tests, Hemoccult is the most popular and the best studied. Sensitivity and specificity are moderate. False negative reactions are due either to the test itself (high level of sensitivity, loss of reactivity) or to the tumour (intermittent or insufficient bleeding, localization etc...). False positive reactions are due to the interference of a high peroxidase diet. The Hemoccult test is nevertheless interesting because it is simple, cheap and well accepted. Detected cancers tend to be at a relatively early stage: 60 to 80% of Dukes A or B. New methods--Hemoquant and immunochemical methods--are being developed. Their high degree of sensitivity and specificity for human hemoglobin makes them too complex and too costly. Whatever the efficacity of a test to detect occult blood may be, we must know whether the screening of an asymptomatic population could decrease the mortality rate of colorectal cancer, before using it on a large scale. Only long-term controlled trials can give us an answer in the future. PMID- 2979055 TI - Monoclonal antibody-mediated targeting of alkylating agents for the treatment of cancer. PMID- 2979054 TI - Experimental application of target-specific immunoconjugates containing daunomycin as the cytocidal component. PMID- 2979056 TI - Synthesis of antibody conjugates with cobra venom factor using heterobifunctional cross-linking reagents. PMID- 2979057 TI - Immunotoxin preparation and testing in vitro. PMID- 2979058 TI - Radioiodinated monoclonal antibodies in the imaging and treatment of human renal cell carcinoma xenografts in nude mice. PMID- 2979059 TI - The use of anthracycline-antibody complexes for specific antitumor activity. PMID- 2979060 TI - Site-specific modification of monoclonal antibodies. Studies of 111indium-labeled antibodies using nude mouse xenograft systems. PMID- 2979061 TI - The development of copper-67-labeled porphyrin-antibody conjugates. PMID- 2979063 TI - KS1/4-DAVLB, a monoclonal antibody-vinca alkaloid conjugate for site-directed therapy of epithelial malignancies. PMID- 2979064 TI - Synthesis and testing of antibody-antifolate conjugates for drug targeting. PMID- 2979062 TI - Preparation and use of DTPA-coupled antitumor antibodies radiolabeled with yttrium-90. PMID- 2979065 TI - Cardiothoracic anesthesia: the subspecialty speaks out. PMID- 2979066 TI - Induction of anesthesia with fentanyl or fentanyl plus etomidate in high-risk patients. AB - Anesthetic doses of fentanyl (46 +/- 1.3 micrograms/kg) and oxygen (group I) were compared to a moderate dose of fentanyl (3 to 4.7 micrograms/kg; mean = 3.54 +/- 0.1 micrograms/kg) + etomidate (0.4 mg/kg) intravenously (IV) (group II) during the anesthetic induction-endotracheal intubation sequence to evaluate hemodynamic changes and the incidence of side effects in 23 New York Heart Association class III and IV patients. Chest wall rigidity only occurred in group I (27%), and pain on injection (8%) and myoclonus (25%) only in group II. Patients in group I experienced transient, small increases in central venous pressure (immediately after induction) and mean pulmonary artery pressure (after tracheal intubation). Patients in group II had small, transient decreases in heart rate, mean arterial blood pressure and cardiac index after induction which returned to baseline levels immediately after tracheal intubation. The data indicate that a modest dose of fentanyl with etomidate is similar to a large (anesthetic) dose of fentanyl in terms of avoiding cardiovascular depression and preventing hemodynamic stimulation during and following the induction-tracheal intubation sequence. Our findings also demonstrate that these doses of fentanyl before etomidate decrease but do not eliminate side effects of etomidate. The results suggest that a modest dose of fentanyl followed by etomidate may be an attractive alternative to high doses of fentanyl in patients with limited cardiovascular reserve, especially when prolonged postoperative respiratory depression secondary to high doses of an opioid is undesirable. PMID- 2979067 TI - Shivering following cardiac surgery: hemodynamic changes and reversal. AB - The effects of shivering on hemodynamics and systemic oxygenation, as well as the effectiveness of therapeutic interventions in decreasing shivering and increasing mixed venous oxygen saturation, were studied. Thirty adult patients undergoing cardiopulmonary bypass with systemic hypothermia were observed for 1 1/2 to 5 hours postoperatively for signs of shivering associated with a simultaneous decrease in oxygen transport. Systemic and pulmonary hemodynamic measurements were made, blood temperature and mixed venous oxygen saturation were monitored via the pulmonary arterial catheter, and oxygen consumption and delivery were calculated. Shivering was graded by a single investigator on scale of 0 to 4, with 0 = no shivering and 4 = continuous violent muscle activity. Therapy was instituted when shivering reached grade 4 or when SvO2 decreased to less than two thirds of its value on arrival in the intensive care unit (ICU). Patients were randomly assigned to receive either morphine sulfate, 5 to 10 mg, or meperidine, 25 to 50 mg intravenously (IV), followed by the other narcotic if the initial drug failed to improve SvO2 or decrease shivering within ten minutes. The end point for successful treatment was a return of SvO2 to within 5% to 10% of its value upon arrival in the ICU or a cessation of shivering that did not recur within 45 minutes. Twenty of the thirty patients shivered sufficiently to decrease SvO2 by more than one third of its initial value, thus requiring pharmacologic therapy. As shivering increased from a score of 0.8 +/- 1.1 to 3.4 +/- 0.9, SvO2 decreased from 74 +/- 6% to 57 +/- 12%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979068 TI - Pulmonary and systemic hemodynamic effects of central venous and left atrial sympathomimetic drug administration in the dog. AB - Systemic vasopressor or inotropic therapy may exacerbate existing pulmonary hypertension; the optimal agent and route of administration in this situation are unknown. The systemic and pulmonary hemodynamic effects of four sympathomimetic agents (dopamine, epinephrine, norepinephrine, and phenylephrine) during central venous and left atrial administration were investigated in the anesthetized dog. All four drugs increased both systemic and pulmonary artery pressures. Dopamine and epinephrine increased cardiac output and reduced systemic vascular resistance. Phenylephrine decreased cardiac output and increased systemic vascular resistance and left atrial pressure. Norepinephrine did not significantly affect cardiac output, systemic vascular resistance, or left atrial pressure. None of the four drugs affected pulmonary vascular resistance. The ratio of systemic to pulmonary vascular resistance decreased with epinephrine and increased with phenylephrine. There were no hemodynamic differences related to the route of infusion for any of the four drugs. However, pulmonary arterial concentrations of the three drugs measured (dopamine, epinephrine, and norepinephrine) were markedly lower during left atrial compared to central venous drug administration; systemic drug concentrations were similar or increased during left atrial compared to central venous drug administration. It is concluded that the relative effects on the systemic and pulmonary circulations differ for the four drugs; rational choice of a vasopressor will depend upon the hemodynamic situation and the desired effect. Left atrial catecholamine administration is effective in decreasing pulmonary arterial drug concentrations and may decrease adverse pulmonary effects in clinical practice. PMID- 2979069 TI - Why publish? PMID- 2979070 TI - Hemodilution affects the pressor response to norepinephrine. AB - A hindlimb preparation of 19 male rats was used to determine the responses to norepinephrine (NE) at different levels of hemodilution at a constant flow, at increased flow and maintained pressure, and after reversal of hemodilution. An initial series demonstrated that the preparation remained stable with unchanged blood pressure, plasma electrolytes, and blood gases; and had reproducible responses to NE throughout the experiment. Two levels of isovolumic hemodilution were achieved by blood/hetastarch exchanges equivalent to 1.5% and 3.0% body weight, which reduced hematocrit to 30.8 +/- 2% and 18.7 +/- 1.5%, respectively. Hemodilution reduced the perfusion pressure from 73.6 mmHg to 58.6 and 55.7 mmHg following 1.5% and 3.0% hemodilution, respectively. A dose-response relationship to NE (0.1, 0.3, 1.0 micrograms/kg) was obtained. The responses to NE were markedly reduced following both 1.5% and 3.0% hemodilution. Retransfusion of packed red blood cells increased the hematocrit from 21.1 +/- 2.2% during hemodilution to 41.2 +/- 2.1% (P less than .01). Concomitantly, perfusion pressure increased from 46.2 +/- 3.6 to 68.3 +/- 6.5 mmHg (P less than .001), close to prehemodilution levels (65.3 +/- 5.3 mmHg). Restoration of hematocrit levels restored responses to NE. In a third series, hemodilution was continued, but the blood flow was increased from 4.0 to 8.3 +/- 0.9 mL/min, and perfusion pressure returned to baseline values. This also restored the pressor responsiveness to NE to near normal. PMID- 2979071 TI - The "cross-fertilization process". PMID- 2979072 TI - Two-dimensional color flow Doppler echocardiography for the intraoperative monitoring of cardiac shunt flows in patients with congenital heart disease. PMID- 2979073 TI - Alterations in somatosensory evoked potentials associated with inadequate venous return during cardiopulmonary bypass. PMID- 2979074 TI - The cardiac patient and noncardiac surgery: the real challenge. PMID- 2979075 TI - Pulmonary artery rupture during cardiopulmonary bypass. PMID- 2979076 TI - Catheter-induced innominate vein perforation: anatomical considerations. PMID- 2979077 TI - Newer cardiotonic agents: implications for patients with heart failure and ischemic heart disease. PMID- 2979078 TI - Pulmonary artery catheters: there are still concerns with their routine use. PMID- 2979079 TI - Case 1--1987. 57-year-old man with dyspnea, dysphagia, and a mediastinal mass. PMID- 2979080 TI - Pro: opioids are preferable to volatile anesthetic drugs for coronary artery surgery. PMID- 2979081 TI - Con: volatile anesthetic drugs are preferable to opioids for coronary artery surgery. PMID- 2979082 TI - Determination of the pulmonary capillary wedge position in patients with giant left atrial V waves. AB - Thirteen patients with giant left atrial V waves during preoperative cardiac catheterization were admitted into the study group. While awake and breathing spontaneously, simultaneous recordings of electrocardiographic leads II and V5, radial arterial traces, and pulmonary arterial or pulmonary capillary wedge traces were obtained. Measurements were made on four consecutive cardiac cycles in the unwedged and wedged positions for the following intervals: Q wave to the radial arterial upstroke (220 +/- 20 milliseconds) and peak (360 +/- 10 milliseconds), Q wave to the pulmonary arterial upstroke (170 +/- 20 milliseconds) and peak (350 +/- 20 milliseconds), Q wave to the V wave upstroke (280 +/- 20 milliseconds) and peak (570 +/- 20 milliseconds), and QT interval (420 +/- 20 milliseconds). These findings indicate that the radial arterial and pulmonary arterial upstrokes and peaks occur nearly simultaneously. Upon wedging, the V wave upstroke occurs significantly later in the cardiac cycle (P less than .05) compared with the pulmonary arterial upstroke, and the V wave peak occurs significantly later compared with both the pulmonary arterial and the radial arterial peak (P less than .05). A rapid, simple beat-to-beat method for differentiating pulmonary arterial from pulmonary capillary wedge positions in the presence of giant left atrial V waves is the superimposition of the pulmonary arterial trace on the radial arterial trace. When a wedge position is attained, there is an immediate rightward shift in the upstroke and peak of the pulmonary arterial pressure trace, which can be easily identified by observing the relationship between the pulmonary arterial and systemic arterial traces. PMID- 2979083 TI - The influence of adenosine triphosphate on left ventricular function and blood flow distribution during aortic crossclamping in dogs. AB - To evaluate the influence of adenosine triphosphate (ATP)-induced vasodilation on myocardial performance and blood flow during aortic crossclamping (XC), ten dogs were instrumented to measure left ventricular (LV) pressure and dimensions. Regional LV function was assessed from the percentage of systolic shortening, whereas the slope of the linear regression of the LV end-systolic pressure diameter relationship was used as an index of overall contractility. The regional blood flow distribution was measured from sequential injections of radioactive microspheres. Following XC, systemic arterial pressure proximal to the clamp (SAPa), LV end-diastolic pressure (LVEDP), LV end-systolic meridional wall stress (WS), and central venous pressure (CVP) increased significantly, whereas the cardiac index (CI) and heart rate did not change. After 30 minutes of ATP infusion (1 mg/kg/min) SAPa, LVEDP, WS, and CVP returned to control levels, CI increased significantly compared with XC alone, and vascular resistance fell below the control level. ATP produced a threefold increase in myocardial blood flow and shifted the intramural distribution in favor of the endocardial layer. In conclusion, our investigation of the effect of ATP on aortic XC in a canine model showed the drug to produce a smooth, predictable, and rapid reduction in left ventricular preload and afterload. This was accomplished with minimal changes in distal organ perfusion, some improvement in measured cardiac performance, and a large increase in myocardial blood flow. PMID- 2979084 TI - Right ventricular ischemia diagnosed by an esophageal electrocardiogram and right atrial pressure tracing. PMID- 2979085 TI - Perioperative right ventricular ischemia. PMID- 2979086 TI - Anesthesia for neonatal orthotopic cardiac xenograft. PMID- 2979087 TI - Deep hypothermic circulatory arrest: a review of pathophysiology and clinical experience as a basis for anesthetic management. PMID- 2979088 TI - Case 2--1987. 59-year-old man with mitral stenosis secondary to rheumatic fever presented for mitral valve replacement. PMID- 2979089 TI - Pro: pulsatile flow is preferable to nonpulsatile flow during cardiopulmonary bypass. PMID- 2979090 TI - Con: nonpulsatile flow is preferable to pulsatile flow during cardiopulmonary bypass. PMID- 2979091 TI - Monitoring for right ventricular ischemia: is it necessary? PMID- 2979092 TI - The influence of positive end-expiratory pressure on intrapericardial pressure and cardiac function after coronary artery bypass surgery. AB - The hemodynamic effects of positive end-expiratory pressure (PEEP) were studied in coronary artery bypass patients by recording intrapericardial and intracardiac pressures, measuring cardiac output by thermodilution, and determining left ventricular volumes by nuclear radiography. An elevation of PEEP to 5, 10, and 15 cm H2O led to a decrease in cardiac output (15% decrease at PEEP 15) as intrapericardial pressure increased and transmural left atrial pressure decreased. Modest volume loading (an increase in left atrial pressure of 3 mm Hg) greatly attenuated the deleterious effects of 15 cm H2O PEEP. There was an excellent correlation between pulmonary capillary wedge pressure and left atrial pressure at PEEP 0 and 5 (r = .85 and r = .83). This correlation was not nearly as reliable at PEEP 15 (r = .54). A predictable increase in intrapericardial pressure was observed as PEEP was applied in these patients. The magnitude of this increase can be estimated by multiplying the change in PEEP (in cm H2O) by 0.4 to estimate the change in intrapericardial pressure (in mm Hg). Using this estimation as a guide, modest volume loading can be used to maintain transmural filling pressures (and cardiac output) when PEEP is used after coronary artery bypass surgery. PMID- 2979093 TI - Artificial organs in the thoracic cavity. PMID- 2979094 TI - Myocardial ischemia during cardiovascular surgery as detected by an ST segment trend monitoring system. AB - The incidence of intraoperative myocardial ischemia was determined in 312 patients undergoing cardiovascular surgery using ECG recordings obtained from a prototype system that trends ST segment changes. Prior to cardiopulmonary bypass, ischemic ECG changes were observed in 11.9% of patients, the incidence being lower during coronary artery bypass grafting (CABG) procedures (8.1%) than in patients undergoing repeated CABG (23.5%), valve replacement (25.9%), or concomitant valve replacement and CABG (35.3%). The occurrence of intraoperative myocardial ischemia was statistically greater in patients with a history of hypertension, two or more previous myocardial infarctions, or kidney disease. The incidence of ischemia was also significantly greater in patients with left ventricular end-diastolic pressures of 15 mm Hg or higher at cardiac catheterization. The incidence during induction was significantly reduced by the addition of hypnotics to narcotics, and, during maintenance, by the addition of an inhalational agent. The majority of ischemic events were temporally related to some hemodynamic disturbance, and many coincided with surgical manipulation. The incidence of intraoperative myocardial ischemia was substantially less than that reported in comparable studies. Since the incidence decreased significantly following clinical acceptance of the prototype system, the authors conclude that such capability increases sensitivity to small ST segment changes. This heightened awareness and prompt remedial action reduced the incidence of ischemia during CABG to 6% during the final 2 years of the study. PMID- 2979095 TI - Hematologic effects of cardiac and noncardiac surgery. AB - The intraoperative and postoperative changes in number and type of WBCs in patients undergoing cardiac surgery were studied. These changes were then compared with those that occurred in patients undergoing four noncardiac surgical procedures (abdominal vascular reconstruction, thoracotomy, cholecystectomy, and carotid thromboendarterectomy). Both cardiac surgery and abdominal vascular surgery resulted in a marked increase in bands and decrease in lymphocytes. Thoracotomy and cholecystectomy resulted in similar but smaller changes. Carotid thromboendarterectomy did not produce hematologic changes. We conclude that the hematologic changes that occur with cardiac surgery are primarily a result of the stress and trauma of major surgery rather than a result of cardiopulmonary bypass itself. PMID- 2979096 TI - Protection from coronary air embolism by a perfluorocarbon emulsion (FC-43). AB - The protective effects of a perfluorocarbon emulsion (FC-43) against the physiological sequelae of selective coronary air embolism (CAE) were examined in dogs. Animals were randomly assigned to two groups: group 1 received 20 mL/kg of 6% hetastarch in 0.9 normal saline solution with 280 mg/L of CaCl2; group 2 received 20 mL/kg of FC-43 (Oxypherol, Alpha Therapeutic Corp, Los Angeles). Infusion time for both groups was 30 minutes. Following a ten-minute stabilization period, a 0.02-mL/kg bolus of air was injected into the left anterior descending coronary artery distal to the first branch. Surface and epicardial ECGs, systemic BP, pulmonary artery pressure, central venous pressure, left ventricular pressure, left ventricular end-diastolic pressure (LVEDP), dP/dtmax, and Vmax were recorded continuously. Cardiac output (by thermodilution technique), myocardial creatine phosphokinase levels and the lactic acid concentration were measured. The study revealed a significantly lower systemic mean arterial pressure (MAP), LVEDP, left ventricular stroke work index (LVSWI), and right ventricular stroke work index (RVSWI) in the group receiving the FC-43 prior to receiving CAE. Possible causes of the differences are a species-specific direct vasodilating effect of FC-43 or histamine release. Following CAE, myocardial function as measured by dP/dtmax, Vmax, and MAP was lower in group 1 during the first five minutes following CAE. ECG tracings showed all of the ten group 1 dogs had ST-T changes (greater than 2 mm), and six of the ten had ventricular dysrhythmias or conduction defects. Only one of the group 2 dogs showed ST-T wave changes (P = .0006, Fisher's exact test), and none had dysrhythmias (P = .005, Fisher's exact test).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979097 TI - Mechanical heart implantation as a bridge to transplantation: two case reports. PMID- 2979098 TI - Treatment of intraoperative acute right heart failure by a right ventricular assist device. PMID- 2979099 TI - Systemic vascular response to anesthetic induction in an artificial heart patient with fixed cardiac output. PMID- 2979100 TI - Treatment of low cardiac output after aortocoronary artery bypass surgery using a combination of norepinephrine and amrinone. PMID- 2979101 TI - High cardiac output measurements in a patient with congestive heart failure. PMID- 2979102 TI - Case 3--1987. 7-year-old girl with an atrial septal defect for a tonsillectomy and adenoidectomy. PMID- 2979103 TI - Pro: strong inotropes (ie, epinephrine) should be drugs of first choice during emergence from cardiopulmonary bypass. PMID- 2979104 TI - Con: epinephrine and norepinephrine are the inotropes of choice: an opposing view. PMID- 2979105 TI - To use or not to use ATP during aortic crossclamping--that is the question. PMID- 2979106 TI - Heparin: new information on an old drug. PMID- 2979107 TI - Predicting the pharmacodynamics of heparin: a clinical evaluation of the Hepcon System 4. AB - The magnitude of the anticoagulation response to heparin (heparin responsiveness) varies substantially from patient to patient. Identifying extremes of sensitivity and resistance prior to intravenous administration of heparin would facilitate anticoagulation for cardiopulmonary bypass (CPB). The performance of the Hepcon System 4 (HemoTec, Inc, Englewood, CO), an instrument designed for that purpose, was tested. Using nonheparinized blood samples from 157 patients scheduled for surgery requiring CPB, this device performed activated coagulation times (ACT) with three different concentrations of in vitro heparin. After determining each patient's in vitro heparin response, the heparin dose predicted to produce ACT values of 480 seconds (group 1, N = 77) or 600 seconds (group 2, N = 80) was administered. Five minutes later each patient's ACT was determined with the Hemochron method (International Technidyne, Inc, Edison, NJ). Simultaneously, several other variables that might predict heparin responsiveness were investigated. When compared with the observed ACT, the Hepcon System 4 inadequately predicted the response. There was considerable scatter in this comparison, but most frequently the in vitro method substantially underestimated the in vivo heparin dose requirement. Heparin responsiveness decreased significantly with high platelet counts and advanced age, but was unaffected by the initial hematocrit, ACT, partial thromboplastin time, or preoperative heparin therapy. Previous investigations have not identified a relationship between advanced age and reduced heparin responsiveness. Combining the Hepcon heparin dose-response in vitro method with the other parameters evaluated, stepwise regression could only account for 39% of the observed variability in heparin responsiveness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979108 TI - Absence of prostaglandin changes associated with protamine administration in patients with pulmonary hypertension. AB - In patients with pre-existing pulmonary hypertension, severe pulmonary vasoconstriction has been observed following protamine administration. Thromboxane A2, a potent vasoconstrictor, is capable of producing increases in pulmonary vascular resistance, and animal studies suggest that heparin-protamine complexes stimulate thromboxane A2 synthesis. This study assessed the effect of protamine administration on hemodynamics and on plasma thromboxane A2 and its biologic antagonist, prostacyclin, by serial measurement of the stable metabolites, thromboxane B2 and 6-keto-prostaglandin F1 alpha, respectively. Ten adults with pulmonary artery hypertension undergoing elective mitral valve replacement were studied. After termination of cardiopulmonary bypass, baseline hemodynamic measurements were obtained and arterial blood for prostanoid analysis was sampled. Hemodynamic and prostanoid measurements were obtained 5, 10, 15, and 30 minutes after the protamine infusion began. Prostanoid levels were performed by double antibody radio-immunoassay. No significant hemodynamic changes occurred and no significant changes in prostanoid levels were observed. It is concluded that in patients with pulmonary hypertension, heparin-protamine complexes do not consistently raise circulating thromboxane levels, and the relationships among prostanoids, pulmonary hypertension, and systemic hypotension are still not clear. PMID- 2979109 TI - Coronary artery bypass grafting within 24 hours after intracoronary streptokinase thrombolysis. AB - Little specific information currently exists describing the management of patients with an evolving acute myocardial infarction (AMI) treated with direct intracoronary infusion of streptokinase (SK) followed by emergency coronary artery bypass grafting (CABG). A total of 194 patients with an evolving AMI underwent emergency coronary artery angiography with infusion of SK. Thirty-four of these patients with partial restoration of orthograde blood flow in the infarct-related coronary artery (as determined by clinical and objective evidence of myocardial salvage) were referred for emergency CABG. Problems related to the surgical and anesthetic care of these high-risk patients involved: (1) management of resuscitation of patients with AMI, (2) SK-induced coagulopathy and ongoing thrombolysis, and (3) timely CABG to preserve myocardial salvage. To highlight comparisons of SK-CABG management, data regarding 34 consecutive patients having routine non-SK-CABG surgery were collected simultaneously during the study. Data collected retrospectively included: anesthetic drug summaries, time frame of events from admission to the emergency room until commencing bypass, use of invasive monitoring and hemodynamic assist devices, induction complications, operative complications, coagulation derangements, volume replacement, and blood loss. Results revealed no deaths up to 24 hours postoperatively in the 34 emergency SK-CABG patients, even though complications were frequent intraoperatively. Furthermore, there were no statistically significant differences in SK patients v non-SK patients in blood lost, banked blood and cell saver blood administered, or platelet transfusions. However, in comparison to the non-SK-CABG patients, the SK patients received significantly larger amounts of fresh frozen plasma, cryoprecipitate, and aminocaproic acid. PMID- 2979110 TI - Myocardial metabolism, catecholamine balance, and left ventricular function during coronary artery surgery: effects of nitroprusside and nifedipine. AB - The effects of nitroprusside and nifedipine on myocardial oxygen consumption (MVO2), catecholamine release, and left ventricular (LV) function (using 2D transesophageal echocardiography) were compared. Thirty-seven patients undergoing coronary artery surgery, anesthetized with fentanyl, 100 micrograms/kg, were studied. All had good LV function and had been receiving long-term oral beta blocking therapy. Patients were randomly allocated to one of three groups. Group C (n = 12) received no vasodilator and served as control. Group S (n = 13) received nitroprusside at an initial rate of 1 microgram/kg/min. Group N (n = 12) received nifedipine at an initial rate of 0.7 microgram/kg/min. Baseline measurements were obtained ten minutes after intubation. Vasodilator therapy was then started in groups S and N. Infusion rates were adjusted to maintain systolic blood pressure (SBP) between 80% and 120% of baseline values. Additional measurements were made ten minutes after the start of the infusion, ie, before surgery (in group C immediately before surgery), and after sternotomy when the pericardium was opened. The mean (+/- SD) total dose requirements were 1.9 +/- 0.5 micrograms/kg/min for nitroprusside and 1.1 +/- 0.2 micrograms/kg/min for nifedipine. The mean (+/- SD) total infusion time was 31 +/- 5 minutes for nitroprusside and 32 +/- 11 minutes for nifedipine. After sternotomy, heart rate increased in all groups. At this time arterial blood pressure and systemic vascular resistance (SVR) increased in group C. SVR was decreased after the first ten minutes of nitroprusside infusion and after sternotomy in group S. Coronary sinus blood flow, MVO2, and myocardial norepinephrine release increased in group N, but not in groups C or S. After sternotomy, LV percentage area reduction increased in groups S and N, but not in group C. In group N there was a significant correlation (r = 0.65; P less than .05) between the increases in MVO2 and LV percentage area reduction, an estimate of myocardial function. Lactate production occurred in two patients in group C after sternotomy. This was not associated with ECG changes, but in one patient regional wall motion abnormalities developed. No evidence of myocardial ischemia was observed in groups S and N. However, in contrast to nitroprusside, the use of nifedipine was associated with increases in MVO2, myocardial norepinephrine release, and inotropy. PMID- 2979111 TI - Continuous measurement of intracardiac and pulmonary blood flow velocities with transesophageal pulsed Doppler echocardiography: technique and initial clinical experience. AB - Pulsed Doppler techniques have become well established adjuncts to conventional echocardiography in the noninvasive diagnosis of various cardiac malfunctions. Disadvantages of the transthoracic approach, such as inaccessibility and instability of the probe position, limit the continuous application of pulsed Doppler echocardiography during surgery. This study presents a new technique using the transesophageal approach that combines pulsed Doppler measurements with two-dimensional echocardiographic imaging (TDE). The first intraoperative experience with this new enhancement to two-dimensional transesophageal echocardiography (TEE) showed that this relatively noninvasive technique is a safe method allowing constant monitoring of cardiac and pulmonary blood flow velocities. The simultaneous high-resolution two-dimensional imaging facilitates spatial orientation and placement of the sample volume, as well as continued control of the sampling location. The typical flow velocity patterns in standard TEE views are described. The mitral valve and pulmonary artery offer particularly favorable conditions for continuous high-quality TDE measurements. It is concluded that the new technique may further increase the value of TEE to clinicians in the perioperative period. PMID- 2979112 TI - Anesthetic management for cardiac transplantation in North America--1986 survey. AB - Cardiac transplantation has become an established part of the therapy of end stage heart disease. The number of cardiac transplants performed, as well as the number of centers performing them, has increased dramatically in the past 2 years. A paucity of literature on the anesthetic management of patients undergoing cardiac transplantation prompted this survey of 46 United States and Canadian institutions. The report summarizes the perioperative anesthetic management of a total of 1,273 transplant recipients in 34 institutions. Generally, similar anesthetic techniques and agents were used. One notable exception was the percentage of institutions using perioperative pulmonary artery catheter monitoring. As determined from this survey, right ventricular failure remains the leading cause of inability to terminate cardiopulmonary bypass in this patient population. Further, in surveyed institutions, cardiac transplantation expends more physician as well as hospital resources per patient than coronary artery bypass surgery. PMID- 2979113 TI - Malignant esophago-respiratory tract fistulas: anesthetic considerations for exclusion procedures using esophageal bypass. PMID- 2979114 TI - Cardiopulmonary bypass interference with dantrolene prophylaxis of malignant hyperthermia. PMID- 2979116 TI - Cardiothoracic anesthetic techniques from the Far East. PMID- 2979115 TI - Successful resuscitation of a bupivacaine-induced cardiac arrest using cardiopulmonary bypass and mitral valve replacement. PMID- 2979117 TI - Reoperation for coronary artery bypass grafting: anesthetic challenge. AB - The problem of caring for patients undergoing reoperative coronary revascularization is one that cardiac anesthesiologists will face with increasing frequency. Many thousands of CABG procedures continue to be performed annually with ever-increasing survival rates. Consequently, the population at risk for reoperative CABG is growing, while surgical intervention necessarily follows apace. As one recent long-term, retrospective study showed, patients surviving 12 years after CABG have a reoperative rate of 17.3%. Physicians caring for these patients must recognize that they are not seeing patients with routine CAD, but with a different entity: coronary graft disease (CGD). These patients with CGD are different in many ways from those with native CAD, and these differences must be taken into account when planning for their perioperative care. Cardiologists have strived to check the growth of CGD by aggressive emphasis on modification of coronary risk factors such as tobacco use, hypertension, and hyperlipidemia. In addition, recent interest has been focused on a pharmacologic approach via the platelet-prostaglandin system. Surgeons have also attempted to reduce the incidence of CGD by recognition that significantly improved long-term patency rates can be achieved by the use of the internal thoracic artery as a bypass conduit. Consequently, an expanded role for this vessel in the form of free, sequential, and bilateral ITA grafting is currently being advocated as a surgical solution to the problem of CGD. In contrast, the anesthesiologist probably has little to add to the prevention of CGD, but may be able to contribute to a favorable outcome at reoperation. The medical variables and preoperative characteristics that make reoperative CABG patients different from those presenting for primary CABG should be recognized. A firm appreciation of the nature of graft disease, as well as the surgical intricacies required for correction, can only serve to improve the care offered during these often complex operations. Aggressive, invasive hemodynamic monitoring, constant vigilance for signs of early ischemia, and preparedness for prebypass hemorrhage and postbypass ventricular dysfunction should be made. Furthermore, if anesthesiologists are to contribute to an improved outcome in these patients, strategies must be developed to attenuate cerebral and myocardial damage resulting from hemorrhage and atheroembolic catastrophies that appear to be frequent complications in these challenging surgical patients. PMID- 2979119 TI - Pro: regional anesthesia is preferable to general anesthesia for carotid artery surgery. PMID- 2979118 TI - Case 5--1987. 45-year-old woman develops acute left ventricular ischemia and dysfunction after subxiphoid drainage of a pericardial tamponade. PMID- 2979120 TI - Con: carotid endarterectomy: general is safer than regional anesthesia. PMID- 2979121 TI - Pulmonary artery rupture during cardiopulmonary bypass. PMID- 2979122 TI - Intravenous procaine is a useful addition to balanced anesthesia for thoracic surgery. PMID- 2979123 TI - Pro: proper positioning of a double-lumen endobronchial tube can only be accomplished with endoscopy. PMID- 2979125 TI - The safety of femoral artery cannulation. PMID- 2979124 TI - Con: proper positioning of a double-lumen endobronchial tube can only be accomplished with the use of endoscopy. PMID- 2979126 TI - A helpful tip for insertion of pulmonary artery catheters. PMID- 2979127 TI - The anesthetic management of adult patients undergoing noncardiac intrathoracic surgery. PMID- 2979128 TI - Alfentanil in infants and children with congenital heart defects. AB - The use of an alfentanil infusion as a supplement to a nitrous oxide-halothane anesthetic and the pharmacokinetics of alfentanil were evaluated in infants and children undergoing surgery for correction of congenital heart defects. Eleven patients, six infants and five children, were studied. Anesthesia was induced with nitrous oxide-halothane and pancuronium, 0.15 mg/kg. After intubation, anesthesia was maintained with nitrous oxide-oxygen and halothane to a maximum inspired concentration of 0.6%. After administration of atropine, 20 micrograms/kg, alfentanil, 20 micrograms/kg, was given, followed by a continuous infusion of 1 microgram/kg/min, which was stopped after closure of the sternum. Supplemental boluses of alfentanil, 5 micrograms/kg, were given when, during surgery, blood pressure and/or heart rate increased more than 20% above control values. At the end of surgery, after antagonism of residual neuromuscular blockade, the patients were extubated. Arterial blood samples were collected at regular intervals during surgery and for six hours thereafter for determination of alfentanil plasma concentrations by gas chromatography. Pharmacokinetic data were calculated using the method of residuals and noncompartmental moment analysis. Although atropine was administered, heart rate decreased significantly (2.5% to 15%) in all infants after administration of alfentanil. In the older children, blood pressure decreased 10% to 35%. In the period before bypass, three infants and four children needed supplemental boluses of alfentanil. During and after bypass, anesthesia was adequate. All patients could be extubed within 34 minutes of stopping the alfentanil infusion. Naloxone was not required in any patient, and postoperative respiratory depression did not occur. In the infants and children, total plasma clearance was 8.2 +/- 2.2 mL/kg/min and 6.3 +/- 0.8 mL/kg/min, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979129 TI - Anesthetic implications for the management of patients with acute myocardial infarction: a matched cohort study of patients undergoing emergency myocardial revascularization. AB - Emergency coronary artery bypass grafting (CABG) is advocated as a treatment of acute myocardial infarction (AMI). To attempt to define anesthetic management problems in this patient group, a retrospective study was conducted comparing the perioperative courses of 23 patients undergoing emergency CABG during AMI with 23 elective patients, individually matched for gender, operating surgeon, ejection fraction, and aortic crossclamp time. The 23 AMI patients were anesthetized 5.98 +/- 3.0 (range 1.5 to 11.0) hours after the onset of chest pain. Anesthetic agents were similar for both groups. Induction of anesthesia was well tolerated by AMI patients. Tolerance of cardioplegic arrest was impaired in the AMI group as evidenced by the sharp increase in frequency of inotropic support required to discontinue bypass in the AMI group compared to elective patients (12/23 v 3/23; P less than .005). Fifteen AMI patients who received preoperative streptokinase had greater postoperative bleeding. Three AMI patients died postoperatively. The number of patients requiring prolonged postoperative ventilation and extended ICU care was higher in the AMI group. It is concluded that patients undergoing emergency CABG during AMI represent a greater risk than elective patients. They have a higher incidence of myocardial dysfunction following cardioplegic arrest during bypass. Those who receive preoperative thrombolytic therapy exhibit greater bleeding tendencies. PMID- 2979130 TI - Oxygenation and hemodynamic changes during one-lung ventilation: effects of CPAP10, PEEP10, and CPAP10/PEEP10. AB - The effects of 10 cm H2O positive end-expiratory pressure (PEEP10), 10 cm H2O continuous positive airway pressure (CPAP10), and their combination (CPAP10/PEEP10) on oxygenation and hemodynamics were studied in 20 patients undergoing one-lung ventilation (OLV) with 50% nitrous oxide, isoflurane, and oxygen. Compared to OLV alone, CPAP10 and CPAP10/PEEP10 significantly increased PaO2 (from 80 +/- 6 to 125 +/- 11 and 137 +/- 17 mmHg, respectively); increased SaO2 (from 93.9 +/- 0.8 to 97.1 +/- 0.5 and 97.0 +/- 0.6%, respectively); and decreased Qs/Qt% (from 36.4 +/- 1.6 to 26.2 +/- 2.0 and 23.2 +/- 2.0%, respectively). Although not statistically significant, PEEP10 caused an increase in PaO2 (to 105 +/- 12 mmHg) and a decrease in Qs/Qt% (to 27.6 +/- 2.1%), which are of clinical significance. However, CPAP10/PEEP10 caused a significant decrease in cardiac output (from 4.50 +/- 0.26 to 3.83 +/- 0.22 L/min), stroke volume (58.6 +/- 3.0 to 52.8 +/- 2.9 mL/beat), and oxygen delivery (653 +/- 39 to 590 +/- 38 mL/min). Application of CPAP10, PEEP10 or their combination had no significant effect on heart rate, arterial, pulmonary arterial, mean pulmonary capillary wedge or central venous pressures, systemic or pulmonary vascular resistances, or mixed venous oxygen saturation. Overall, CPAP10 had the most beneficial effect on oxygenation and hemodynamics during OLV with 50% N2O, isoflurane and oxygen. PMID- 2979131 TI - Isoflurane elimination via a bubble oxygenator during extracorporeal circulation. AB - It has been suggested that inhalational anesthetics should be discontinued at least 15 minutes prior to termination of extracorporeal circulation (ECC) to avoid myocardial depression. However, data regarding elimination of inhalation agents via a bubble oxygenator from hypothermic, hemodiluted patients have not been previously reported. The washout of isoflurane (ISF) from ten cardiac surgical patients using mass spectrometry was studied. The mean baseline oxygenator exhaust concentration of ISF was 0.85% prior to termination of ECC. Oxygenator concentration of ISF decreased to less than 0.05% in 8.8 +/- 2.5 minutes. Eight of ten patients had ISF washout curves best characterized by a one compartment model, with a mean time constant of 1.94 minutes. Therefore, 95% washout of ISF should occur in 5.8 minutes (three time constants). It is suggested that ISF may be used closer to the termination of ECC than previously recommended without fear of significant myocardial depression. PMID- 2979132 TI - Modification of myocardial ischemic injury: a concentration response study of glucocorticoid supplementation during reperfusion. AB - Reperfusion of the ischemic myocardium is routinely done during cardiac surgery and in the catheterization laboratory after acute regional ischemia. While reperfusion of the ischemic myocardium is necessary in order to regain full functional and biochemical recovery, the reperfusion by itself may aggravate the ischemic damage. Glucocorticoids have been shown to modify the outcome from ischemic injury in various experimental and clinical situations, but the results are conflicting. This protocol was performed using normothermic ischemic (30 minutes) isolated rat hearts, and postischemic reperfusion (30 minutes) with methylprednisolone sodium succinate (MPSS; 100-500-1,000 mg/L) was studied. All indices of myocardial function and all metabolic variables were significantly reduced after ischemia. MPSS (100 mg/L) improved dP/dTmax (recovery 68 +/- 3% v control 48 +/- 7%) and some of the other indices of left ventricular performance studied. MPSS (100 mg/L) also improved the tissue concentration of adenosine triphosphate (ATP) (13.8 +/- 0.5 mumol/g dry weight v control 11.3 +/- 0.8) and the total adenosine pool (16.8 +/- 0.7 mumol/g weight v control 13.9 +/- 0.8 mumol/g dry weight). MPSS (1,000 mg/L) impaired recovery of myocardial function (dP/dTmax, dP/dtmin, rate-pressure product [RPP]), increased tissue lactate (7.2 +/- 2.0 mumol/g dry weight v control 3.6 +/- 0.8), reduced glycogen (30.6 +/- 2.5 mumol/g dry weight v control 49.8 +/- 3.3), and energy charge (0.848 +/- 0.018 v control 0.890 +/- 0.010). It is concluded from the present experiments that modification of an ischemic injury by glucocorticoids given at the onset of reperfusion is possible, and that an optimal concentration of 100 mg/L exists for MPSS supplementation. MPSS in a concentration of 1,000 mg/L aggravated the reperfusion injury, probably by interference with cellular respiration. PMID- 2979133 TI - Prostaglandin E1 for refractory right heart failure after coronary artery bypass grafting. PMID- 2979134 TI - Mitral regurgitation during withdrawal of mechanical ventilatory support. PMID- 2979136 TI - Transesophageal echocardiography: technique and clinical applications. PMID- 2979135 TI - Intratracheal insufflation combined with intermittent positive pressure ventilation for treatment of terminal respiratory failure in a child: a new technique. PMID- 2979137 TI - Case 1--1988. A 73-year-old man sustains an acute aortic disruption after aortic valve replacement. PMID- 2979138 TI - [Multiple echinococcosis with predominant cardiac localization as the cause of death in sudden deaths]. PMID- 2979139 TI - Nickel and vanadium analysis and cytochemistry of heart muscle and sera under different pathological conditions. PMID- 2979140 TI - [New aspects of forensic judgement of indications of open heart surgery]. PMID- 2979141 TI - Diagnostic value of deletion of myoglobin from myocardial fibres in acute cardiac failure. PMID- 2979142 TI - [The sudden death of a 4 1/2-year-old child caused by hypertrophic obstructive cardiomyopathy owing to an abdominal neuroblastoma]. PMID- 2979143 TI - [The binding affinity of tissue proteins depending on postmortem autolysis]. PMID- 2979144 TI - Post mortem damage of corpses by dogs and difficulties involved in diagnosis. PMID- 2979145 TI - [An original procedure for ballistic identification]. PMID- 2979146 TI - [An attempt to explain the mechanism of the appearance of nonspecific reactions in bones subject to putrefaction]. PMID- 2979147 TI - [Criminal dismemberment of the corpse]. AB - In the period from 1959 to 1984 the Institute of Legal Medicine in Hamburg had to investigate 27 cases of criminal dismemberment. With regard to all legal autopsies the average frequency was about 1:500. A distinct increase of the number of cases is noticeable within the last years; i.e., 1984 there were 6 victims. Our study enclosed the autopsy findings, the police investigations, and the available court records. 11 cases were classified as defensive and 16 cases as offensive dismemberment of bodies (among these: 2 cases of dismemberment after stealing the corpse and 2 cases of killing the victims by decapitation). One perpetrator had 3 victims, twice there was a perpetrator with 2 victims. The victims were of all ages, but mainly between 40 and 50 years old. 22 of them were female and 5 male. 16 perpetrators could be identified, 14 of them were male and 2 female; all of them were older than 30 years, mostly between the age of 40 and 50. The psychiatric experts classified 11 of the perpetrators as "normal," 3 as abnormal, and 2 as psychotic. More details concerning victims, perpetrators, used instruments for killing and dismembering, elimination and identification of the dismembered bodies and judicial aspects are given in this paper. PMID- 2979148 TI - Immuno-histochemical reactions with monoclonal antileukocyte antibodies in vital injuries. PMID- 2979150 TI - [Identification criteria in maternal infanticide]. PMID- 2979149 TI - Studies on the specificity for the grouping of extremely small amounts of bloodstains. PMID- 2979151 TI - A procedure for systematic examination on small pieces of blood-stained thread specimens. PMID- 2979152 TI - Immunohistochemical study of blood group activities in human gastrointestinal tissues in normal and pathological conditions. PMID- 2979153 TI - A study of the identification of human saliva by anti-alpha-amylase. PMID- 2979154 TI - Effect of fixatives on the detection of Y bodies reprint preparations of parenchymatous organs. PMID- 2979155 TI - Sudden death in athletes. PMID- 2979156 TI - Effect of pH values and ion composition of buffer on the detection of Y bodies. PMID- 2979157 TI - Suicide or homicide--biochemical reconstruction of complicated cases of death. PMID- 2979158 TI - How can the safety of sperm stains determination increase? PMID- 2979159 TI - Suicide or homicide? PMID- 2979160 TI - [Immunoelectrophoresis for the expertise of blood stains in forensic medicine]. PMID- 2979161 TI - Application of interference-contrast and polarizing microscopy in detection of traces of biological origin. PMID- 2979162 TI - The proof of gravidity in blood spots with the use of trophoblast-specific beta-1 glycoprotein (TSBG). PMID- 2979163 TI - Comparative investigation of human endomyocardial biopsy. PMID- 2979164 TI - Sudden and unexpected deaths in police custody. PMID- 2979165 TI - Phase transition phenomenon in bodies of persons found dead in bath-tubs. PMID- 2979166 TI - Functional and morphological investigations of the heart in acute experimental hypoxia. PMID- 2979167 TI - A modification of the demonstration of glycogen-dependent phosphorylase activity suitable for autopsy material in the diagnosis of sudden death. PMID- 2979168 TI - Microscopic investigations of right ventricle in sudden coronary death. PMID- 2979169 TI - Cardiac lesions in methamphetamine abusers. PMID- 2979170 TI - Sudden deaths in South London 1960 cf. 1985. PMID- 2979171 TI - [The delimitation of sudden deaths by anatomicopathological examinations]. PMID- 2979172 TI - Fiedler myocarditis as cause of sudden death. PMID- 2979173 TI - [Ischemic cardiopathy--the corroboration of histopathological and thanato enzymological data]. PMID- 2979174 TI - [The medicolegal value of thallium-201 myocardial scintigraphy in assessing the residual damage of closed thoracic injuries]. PMID- 2979175 TI - Epidemiology of sudden unexpected cardiac death of cardiovascular origin in young and middle aged Finns and some preliminary results to apply to post mortem serum and arterial wall lipid analyses to their studies. PMID- 2979176 TI - Sudden unexpected natural death in young persons. PMID- 2979177 TI - Comparative morphological investigations in sudden cardiac death. PMID- 2979191 TI - [Oral rehabilitation in a patient with deep anterior crossbite]. PMID- 2979192 TI - [Human cell transplantation]. AB - To summarize this review: it was written concerning human cell transplantations from various human organs based on already published historical reports. It was limited to only clinical cases for therapeutic treatment. Some of the methods in the article are commonly used, well-established methods found in everyday practice. Others are still in the experimental stages. I think we can try these, or even never methods for other cell groups in the near future. PMID- 2979193 TI - [Recent problems in cell culture systems using normal human cells]. AB - Some remarkable studies of human cell culture have contributed to many basic and applied sciences on "normal human cells"; developments of biological products or physiologically activated substances. In this reviews, some problems concerning in vitro culture systems were discussed and recent advances on the researches of normal human cells were described. PMID- 2979194 TI - [Normal skin cells in vitro]. AB - Major cells in skin are epidermal cells in epidermis and fibroblasts in dermis. These cells can be isolated as a relatively pure population from the tissues using proteases and chelating agents. In this review we describe the way of culture where these two kinds of cells express normal function as they do in vivo. 1) It is important to consider the polarity of epidermal cell membranes in the transport of nutrients and metabolites when the cells are to be cultured in a healthy state in a long period. Epidermals cells expressed their normal polarities when cultured on a porous thin film of collagen and bathed on both sides (apical and basal) in culture media. 2) It is important to consider the interactions of fibroblasts with collagen when normal morphology and physiology are expected to be expressed in the cell in culture. Collagen affected the morphology of the cell and profoundly decreased the rate of DNA synthesis. We present a hypothesis which explains the fibronectin-independent interaction of fibroblasts with collagen. 3) It is important to consider the interactions between fibroblasts and epidermal cells when normal physiology of the skin as a whole is expected to be expressed in vitro, because exchange of information between them control their metabolic activities and functions. In this review, two examples for this exchange are presented: cell growth and collagenolysis. PMID- 2979195 TI - [Culture of human blood vessel endothelial cells--a simple and inexpensive culture method]. AB - The purpose of this review is to introduce a simple and inexpensive culture method for human umbilical blood vessel endothelial cells. The medium used is MCDB-104 supplemented with 10% fetal bovine serum, 70 ng/ml endothelial cell growth factor from new-born bovine brains, 10 ng/ml murine epidermal growth factor, and 100 micrograms/ml heparin. The culture dishes are coated with gelatin. Under these conditions, endothelial cells from human vessels were grown with doubling times of 18-22 hrs and reached saturation densities of 8-12 x 10(4) cells/cm2. To determine the lifespan of the endothelial cells, the cells were serially subcultivated weekly at an inoculum size of 1,000 cells/cm2. Human endothelial cells from umbilical vein and artery were grown for 21 to 37 passages with 55 to 125 population doublings. This culture method seems to be useful for studying cell proliferation and functions of human endothelial cells. PMID- 2979196 TI - [Effects of antipyretics and anticancer drug on human cells at high temperature condition]. AB - The effects of antifebriles and anticancer drug on human vascular endothelial cells (HVE) and several cultured human cells were investigated. The HVE were isolated from umbilical cord veins by enzyme treatment and cultured successively in aerated synthetic medium, RPMI-1640, with 20% preclostrum new born calf serum. The presence of factor VIII antigen in the HVE was determined by enzyme-labeled antibody method. Cell count and protein amount were examined at regular intervals. At 3 hour-expose, sulpyrine was more toxic to the cultured cells than aspirin at 37 degrees C. The cytotoxicity of sulpyrine was markedly enhanced at 40 degrees C than at 37 degrees C. However, there was no enhancement in the cytotoxicity of aspirin at 40 degrees C. Cultured HVE and normal human fetal lung (HAIN-55) cells at 37 degrees C were sensitive to sulpyrine, and their sensitivity of the cells to the drug were markedly enhanced when they were incubated at 41 degrees C. In contrast, sensitivity of malignant human cells (HeLa cells) to sulpyrine was not found at 37 degrees C, however sensitivity of the cells to the drug was manifested at 41 degrees C of incubation. There was no effect of 5-fluorouracil (FU) on the growth of HVE and HAIN-55 cells at 41 degrees C, while HeLa cells showed high susceptibility to FU at the same temperature. The results showed the possibility that normal human cells may be sensitive to antifebrile drugs but not to anticancer drug at ordinary and high temperature, whereas malignant human cell may be susceptible to both antifebrile drugs and anticancer drug at high temperature. PMID- 2979197 TI - Growth and hormonal responsiveness of human endometrial stromal cells in culture. AB - The present review describes and discusses published results on growth and hormonal responsiveness of human endometrial stromal cells in culture. The proliferative potential of serially subcultured cells, that is, the number of cell doublings before cells enter mitotic senescence and cease to divide, was unusually high in stromal cells from several endometrial specimens, a property that may reflect the unique proliferative capacity of human endometrium when compared to other adult tissues. Fluorescent visualization of microfilaments revealed distinct age-related changes in the distribution of cytoskeletal fibers. Addition of ovarian steroids to the culture medium of stromal cells resulted in significant morphologic changes. From comparative studies using different culture media it became evident that medium components remarkably influenced cell morphology during early culture periods in an irreversible manner. Cultured stromal cells yielded interesting results in experiments designed to define the role of polyamines in growth regulation. Proliferation was greatly inhibited when polyamine levels were reduced by specific inhibition of ornithine decarboxylase, the first and rate limiting enzyme in polyamine synthesis which produces putrescine by catalytic conversion from ornithine. The antiproliferative effects were reversed by addition of putrescine to the culture medium. These results clearly establish a causal link between polyamine depletion and growth deficiencies and reveal an essential function of polyamines in stromal cell proliferation. Hormonally regulated parameters in cultured stromal cells include aromatase activity, pregnancy-associated plasma protein-A, 51K secreted protein, prolactin and laminin. The hormonally regulated production of prolactin and laminin, both considered markers of decidualization, together with morphologic changes of stromal cells to decidual-like cells, strongly suggest that human endometrial stromal cells, when subjected to appropriate hormonal stimulation, are capable of differentiating into decidual cells in culture. Cultured stromal cells therefore offer a unique opportunity to examine the complex changes in gene expression associated with decidualization. In addition, in vitro decidualization may prove to be an effective diagnostic tool in certain cases of infertility. Finally, decidualization of cultured stromal cells represents a relevant end point for testing compounds of potential clinical importance, such as synthetic progestins or antifertility drugs. PMID- 2979198 TI - [Cell cycle analysis using the monoclonal antibody Ki-67]. AB - We determined by flow cytometry the proportion of cells in cycle with the monoclonal antibody Ki-67 and also in S-phase after incorporation of bromodeoxyuridine (BrdUrd) with monoclonal antibody to BrdUrd. The monoclonal antibody Ki-67 was useful to detect a nuclear antigen present only in proliferating cells but not expressed in resting (Go) cells. Cell preparation to measure BrdUrd amount incorporated into cellular DNA was difficult but this anti BrdUrd antibody was useful for measuring the rate of DNA synthesis and for the analysis of precious cell kinetics. These antibodies may provide useful information of cell kinetics. PMID- 2979199 TI - [Application of logistic curve fitting to cell-mediated cytotoxicity tests]. AB - Blood mononuclear cells from 6 healthy adults were examined for LAK activity against 16 cell lines derived from human malignant tumors. The cytotoxicity curves obtained from a linear plot of the log of E/T ratio versus the percentage of the target cells killed were found to well fit the logistic curves. As the logistic distribution is known to approximate to the normal distribution, and the K/2 value is the function value for the representative, the K/2 value is reasonably selected as the % lysis value for the lytic unit calculation. Values of the three parameters, K as the maximum cytotoxicity, A as the scale parameter and B as the shape parameter, were analyzed using the experimental data. Eleven targets had three or more significantly fitted curves. Six among them were selected as highly sensitive cell lines to LAK activity, because the average values for K were higher than 80% (coefficients of variation were from 0.02 to 0.11) and analyzed further. The average values for B were relatively constant (range: 1.09-1.56) and the average values for A varied from 3.5 to 20.3 (conefficients of variation were from 0.26 to 0.71). The shape of the curves were nearly the same. These results indicate that the 6 cell lines are useful as the targets for the LAK activity. PMID- 2979201 TI - [Establishment and characterization of human colonic and gastric cancer cell lines]. AB - Two human cancer cell lines, DAIT-6 from a colonic cancer and IT-25 from a gastric cancer, derived from xenografts in nude mice have been established in tissue culture and maintained for over two years. In tissue culture, DAIT-6 cells grew in a monolayered sheet with a population doubling time of about 45.0 hr, and floated or piled up to form small buds above the monolayered surface in relatively confluent cultures. Chromosomal counts ranged from 40 to 108 with a modal number of 59. The cells secreted CEA (1.7 ng/1 x 10(6) cells/24 hr) and CA19-9 (540.5 u/1 x 10(6) cells/24 hr) in spent medium. The IT-25 cells grew in a monolayered sheet with a population doubling time of about 57.8 hr in tissue culture. The IT-25 cells also secreted CEA (0.5 ng/1 x 10(6) cells/24 hr) and CA19-9 (120.0 u/1 x 10(6) cells/24 hr) in spent medium. The xenografts for DAIT-6 and IT-25 in nude mice were histopathologically classified as a moderately differentiated tubular adenocarcinoma and a well differentiated tubular adenocarcinoma, respectively. PMID- 2979200 TI - [Changes in cell cycle and RNA content in cultured cancer cells treated with cis diamminedichloroplatinum (II)]. AB - Recent reports have shown that CDDP interacts with RNA and protein as well as DNA. We studied the alteration of cell cycle, cellular RNA content and the effect of nucleic acid metabolism on cultured cancer cells after treatment with CDDP by flow cytometry and 3H incorporation assay. The alteration of cell cycle was found to be accumulation of cells in after delay S phase in cytostatic concentrations, CDDP inhibited 3H-TdR uptake markedly at this time and 3H-UR uptake earlier. Increase in RNA content accompanied accumulation of cells in G2M phase. This increase was not a specific phenomenon caused by CDDP, because increase in RNA content was also induced by other inhibitors of DNA synthesis. It is more likely that the direct alteration of cell cycle and cellular RNA content due to action of DNA-combined CDDP rather than that of RNA-combined CDDP. PMID- 2979203 TI - [Immunoelectron-microscopic findings of cells concerning immunological recognition]. AB - Recent advance in immunology is evident since the development of monoclonal antibodies and recombinant DNA techniques. Nevertheless, morphological studies in immunology is necessary to understand the mechanisms of immunological reaction in vivo and immunological disease. In this article, morphological findings mainly under immunoelectron microscopy of cells concerning immunological recognition are described. In a paragraph of peripheral bloods, findings of CD3+, CD4+, CD8+, Leu7+ and HLA-DR+ cells are shown with review of recent evidence of immunological recognition and mechanisms of CML. In a paragraph of peripheral lymphoid tissue, morphological changes of B lymphocytes in B cell proliferation are described with changes of surface markers on B cells. In a paragraph of thymus, recent knowledges about immunological tolerance and MHC restriction are reviewed with immunohistochemical findings of the cells in thymus. PMID- 2979202 TI - [Establishment and characterization of human pancreatic adenocarcinoma cell line JHP-1 producing carbohydrate antigen 19-9 and carcinoembryonic antigen]. AB - A new tumor cell line derived from the ascites of a patient with adenocarcinoma of the head of pancreas was established in culture and the nude mouse. The cell line was characterized by the growth with a population doubling time of 22 hr., a high plating efficiency on the plastic surface and a modal chromosome number of 66. The tumorigenicity was proved by the growth in nude mouse and in soft agar. Morphologically the cell line grew as a confluent monolayer with tight adhesion to the plastic surface. Histologically the cell line was epithelial-like in culture and poorly differentiated adenocarcinoma in nude mouse. Ultrastructurally the cell line showed a characteristic pancreatic epithelium. Furthermore, the cell line expressed carbohydrate antigen 19-9 and carcinoembryonic antigen. This cell line, designated JHP-1, has been cultured for at least 100 passages in vitro and maintained for more than 2 years. This cell line would be used as a new model for human pancreatic carcinoma. PMID- 2979204 TI - [Lymphocytes in transplantation immunity--in the relation to HLA antigens]. AB - Allograft rejection, in kidney transplantation, is initiated by recipient's lymphocytes with the recognition of genetically determined antigens, histocompatibility antigens, which are expressed on the graft, followed by killer T cell induction and antibody production against the graft. Recent advances in our knowledge of the human major histcompatibility complex antigens-HLA antigens, which have an important role in transplantation immunity, are herein reviewed briefly and the relation to allograft rejection is discussed. PMID- 2979205 TI - [CD antigens and the differentiation of human lymphocytes]. AB - Many monoclonal antibodies directed against differentiation antigens expressed on human lymphocytes have been produced to study the development and functional difference of human lymphocytes. These monoclonal antibodies were classified to the group of "cluster of differentiation (CD)". We reviewed the functional roles of CD antigens on the activation, proliferation and differentiation of human lymphocytes. PMID- 2979206 TI - [Clinical therapeutic effect of adoptive immunotherapy using IL-2-cultured autologous lymphocytes]. AB - Our method of adoptive immunotherapy (AIT) using autologous IL-2-cultured lymphocytes differs from so-called LAK therapy in several points. We (1) obtain cultured lymphocytes from effusion lymphocytes (EL) or regional lymph-node lymphocytes (RLNL), when possible, rather than peripheral blood lymphocytes (PBL), (2) use crude IL-2 to induce T cell proliferation and to maintain killer activity, (3) use sonicated autologous tumor extract as antigen (Ag) to stimulate proliferation of cytotoxic T cells, and (4) pretreat the patients with local administration of OK-432 before AIT to induce effector cells that act synergistically with transferred killer cells. Surface marker analysis showed that OKT3, IL-2 receptor, Leu 2+15- cells were elevated while Leu 11a and Leu 3+8+ cells were decreased. Culture of RLNL augmented the expression of Leu 3+8- marker. Both of PBL and RLNL responded to Ag, and their auto-tumor killing activities were augmented in about half of the patients while rarely decrease by the addition of Ag. Response rates of patients with pleural effusion due to breast cancer and those with liver metastasis of breast cancer were 94% and 60%, respectively. Moreover, the survival was prolonged in the treated patients with pleural effusion or gastric cancer patients with peritoneal dissemination. PMID- 2979207 TI - [Cytokinetic effects of carboplatin and cisplatin on a human ovarian cancer cell line]. AB - The cytokinetic effects of carboplatin(CBDCA) on a human ovarian cancer cell line(KF-1) were examined by means of cell survival rate and flow cytometry in comparison with cisplatin(CDDP). CBDCA and CDDP exhibited dose dependent cytotoxicity on KF-1, and CBDCA showed compatible cell growth inhibition to that of 15 times concentration of CDDP in comparison with IC50 of 72 hrs after drug addition. From the analysis of cell cycle, CBDCA and CDDP inhibited cell cycle progression at G2 + M phase. CBDCA exhibited G2 + M phase block to that of 15 to 20 times the concentration of CDDP. We suggested that CBDCA had potential therapeutic activity against ovarian cancer, but should be evaluated carefully in the clinical use. PMID- 2979208 TI - [Establishment and characterization of a new SCC antigen producing cell line (HCS 2) from a carcinoma of the uterine cervix]. AB - The cell line designated HCS-2 established from a squamous cell carcinoma of the uterine cervix has been subcultivated 77 times since Nov. 8, 1983. The cultured cells appear epithelial in shape, with a pavement-like arrangement and grow without contact inhibition. In electron microscopy, the cells are characterized by desmosomal cell contacts and a few tonofilaments. The cells are transplanted subcutaneously to nude mice and produce tumor which resembles the original tumor of large cell non-keratinizing squamous cell carcinoma. The growth rate of subculture has increased gradually, and population doubling time of cells at 17th passage was about 65 hours. The chromosome studies show aneuploidy and chromosomal number was mainly from hypertriploid to hypotetraploid range. The modal number of cells at 33rd passage was 81. Specific marker chromosome is not realized. The production of SCC antigen is detected from the cells and the amount of SCC antigen in cultured media was recorded from 1.5 to 2.0 ng per 1 x 10(4) cells for 48 hours. CEA synthesis is also confirmed immunohistochemically. PMID- 2979210 TI - Cyclosporin for Crohn's disease. AB - Eight patients with active uncomplicated Crohn's disease, who were resistant to or intolerant of conventional treatment, were treated for 6 weeks with oral cyclosporin (mean dose 8.2 mg kg-1 day-1). Seven of the eight patients responded to treatment with cyclosporin by symptomatic improvement, weight gain and a return of serum C-reactive protein concentration towards normal. All patients relapsed on stopping cyclosporin. No serious side-effects were encountered. This favourable early experience justifies further trials using cyclosporin for active Crohn's disease. PMID- 2979209 TI - Intragastric pH monitoring in acute upper gastrointestinal bleeding and the effect of intravenous cimetidine and ranitidine. AB - Intragastric pH was measured continuously from 1800 to 1200 hours the following day in 22 duodenal ulcer patients and in eight gastric ulcer patients, all of whom had been admitted as emergencies with acute upper gastrointestinal haemorrhage. The effects of intravenous cimetidine or ranitidine were compared with no treatment. In patients with duodenal ulcer, median intragastric pH was 1.8 (range 1.0-4.9) in the group receiving no treatment. In the cimetidine group (400 mg, 6-hourly, n = 8) median pH was 4.7 (range 1.5-7.7) and after ranitidine (50 mg, 6-hourly, n = 10) it was 3.8 (range 1.2-7.8). The pH remained above 4.0 for 67% of the recording time with cimetidine, 47% with ranitidine and for only 3% with placebo. Intragastric pH in gastric ulcer patients without treatment was higher (median 3.4, range 1.0-6.9) than in duodenal ulcer patients with treatment. Both H2 antagonists raised intragastric pH in patients with gastric ulcer and maintained a gastric pH of greater than 4.0 for at least 50% of the time. Presently recommended i.v. doses of cimetidine and ranitidine do not consistently maintain gastric pH above 4.0 for long periods in patients with peptic ulcer bleeding. PMID- 2979211 TI - Chronic administration of the somatostatin analogue SMS 201-995 does not lead to endogenous antibody formation. AB - Plasma samples of seven patients with gut and pancreatic endocrine tumours who have been on long-term treatment with a long-acting somatostatin analogue (SMS 201-995) were investigated for endogenous antibodies to the peptide by incubation with radiolabelled SMS 201-995. The duration of treatment with the somatostatin analogue was between 9 and 26 months and the dose from 100 to 300 micrograms day 1. In none of the patients could antibodies to SMS be detected. The effect of this somatostatin analogue is unlikely to be impaired by formation of endogenous antibodies, even after long-term treatment. PMID- 2979212 TI - Replacing the acetyl linkage in aspirin with choline and magnesium moieties reduces the occurrence of gastric mucosal injury. AB - The acetyl moiety in aspirin (acetyl salicylic acid: ASA) is considered to play a major part in the pathogenesis of ASA-induced mucosal injury. At equivalent salicylate doses and pH values, the induction of acute gastric mucosal haemorrhagic erosions in rats by ASA and choline magnesium trisalicylate (CMT), a new non-acetylated salicylate, with and without the potentiating damaging effect of taurodeoxycholic acid (TDCA) were compared. Test solutions were administered by per oral intubation to five groups of fasting Sprague-Dawley rats (n = 24). Gastric mucosa were examined after 4 hours and mucosal injury assessed by a lesion-scoring system. The incidence and severity (median lesion scores with quartiles) of the lesions were 83% and 13 (7:20) respectively for ASA (128 mg kg 1) compared with 17% and 0 (0:0) for CMT (128 mg kg-1) (P less than 0.001 and P less than 0.001). TDCA increased mucosal damage to 100% and 29 (20:34) for ASA compared with 30% and 0 (0:4) for CMT (P less than 0.001) and P less than 0.001). Serum salicylate levels (median values of 1.4 for ASA and 1.5 mmol litre-1 for CMT) were not significantly different. It is concluded that replacing the acetyl moiety in ASA with choline and magnesium moieties reduces the ASA-induced mucosal injury, without affecting blood salicylate concentrations. PMID- 2979213 TI - The effects of cigarette smoking on plasma concentrations of gastric antisecretory drugs. AB - Plasma concentrations of cimetidine and ranitidine were measured after oral administration (n = 5 for cimetidine, n = 5 for ranitidine) or intravenous administration (n = 6 for cimetidine, n = 4 for ranitidine) in habitual smokers, once when cigarettes were smoked and again on a separate day when cigarettes were prohibited. After oral administration plasma concentrations of both drugs rose more rapidly and peak plasma concentrations were achieved earlier when cigarettes were smoked. However, plasma concentrations of the drugs subsequent to the peak were significantly lower when cigarettes were smoked. Cigarette smoking had no effect on plasma blood concentrations of either drug when administered intravenously. In eight healthy smokers cigarette smoking increased the gastric emptying of a liquid test meal by 28% compared with non-smoking control rates. In habitual smokers cigarette smoking alters the blood concentrations of antisecretory drugs in a manner which appears attributable to an increase in the rate of gastric emptying. The observed changes in drug disposition may contribute to the loss of gastric secretory inhibition observed in duodenal ulcer patients who are smokers. PMID- 2979214 TI - Acid secretory capacity and plasma gastrin concentration after administration of omeprazole to normal subjects. AB - In a randomized double-blind study, two groups of eight healthy volunteers received either placebo or omeprazole 40 mg o.m. for 14 days. Fasting plasma gastrin concentration and peak acid output in response to a maximal intravenous dose of pentagastrin were measured before, during and after the 14 days of treatment. Omeprazole caused a 68% (mean) decrease in the peak acid output when measured 24 hours after the last dose, with a simultaneous increase in the fasting plasma gastrin concentration. When measured 1, 2, 3 and 8 weeks after cessation of treatment, there was no significant difference in the peak acid output between the two groups. The study demonstrates that there is no increase in the acid production capacity after 2 weeks of treatment with omeprazole. Thus it would appear that the rise in the plasma gastrin concentration during short term treatment with omeprazole does not induce parietal cell hypertrophy or hyperplasia. PMID- 2979215 TI - The effects of intravenous famotidine on pentagastrin-stimulated gastric secretion in man. AB - Eight healthy male volunteers were each studied on four occasions in a 7 hour double-blind, placebo-controlled experiment. Three continuous 6 hour intravenous (i.v.) infusion rates of famotidine, calculated to achieve steady-state plasma concentrations of 10, 30 and 90 ng ml-1, were compared to placebo. Commencing at the third study hour, hourly incremental step-up i.v. infusions of pentagastrin 0.1, 0.2, 0.5, 1.0 and 2.0 micrograms kg-1. hour were administered on each of the four study days. Hourly blood samples were taken for the subsequent determination of plasma famotidine concentration. The three famotidine infusion rates inhibited basal gastric secretion. Pentagastrin-stimulated secretion was inhibited in a dose-dependent manner. The highest dose of famotidine (38.7 micrograms kg-1.hour) inhibited each infusion rate of pentagastrin-stimulated gastric secretion between 92 and 96%. The lowest dose of famotidine (4.3 micrograms kg-1.hour) inhibited pentagastrin 0.1 micrograms kg-1.hour stimulated secretion by 50% but, when stimulated with pentagastrin 2.0 micrograms kg-1.hour, gastric secretion was inhibited by only 25%. This study indicates that the intravenous preparation of famotidine is a potent inhibitor of pentagastrin-stimulated gastric acid secretion. PMID- 2979216 TI - SMS 201-995 treatment and advanced intestinal cancer: a pilot study. AB - The hypothesis that somatostatin, a compound with antitrophic effects on the gastrointestinal tract, may affect beneficially the progression of advanced intestinal cancer has been tested in a small pilot study. Ten patients, four with advanced pancreatic cancer, four with advanced colorectal cancer and two with gastric cancer, were treated with a long-acting analogue of somatostatin, SMS 201 995, 50, 100 micrograms subcutaneously twice daily. There were no clinical, radiological or biochemical indicators of a response to this treatment. There are no indications from this study that hormonal manipulation alters the rate of growth of advanced gastrointestinal cancer. PMID- 2979217 TI - Comparison of calcium channel blocking agents and an anticholinergic agent on oesophageal function. AB - The effects of oral doses of three calcium blockers and an anticholinergic drug on oesophageal function were compared. Nifedipine (20 mg) and hyoscyamine (0.25 mg) significantly reduced lower oesophageal sphincter (LES) pressure and oesophageal contractile pressure. Verapamil (120 mg) and diltiazem (60 mg) had no significant effect on any of the oesophageal variables measured. Oesophageal transit time and oesophageal contractile duration were not affected significantly by any of the agents. Only hyoscyamine significantly prolonged acid clearance time. The combination of nifedipine and hyoscyamine was no more effective in decreasing LES pressure or oesophageal contractile pressure than either agent alone. Either nifedipine or hyoscyamine would appear to be potentially effective for the treatment of oesophago-spastic and other hypertensive motor disorders, but hyoscyamine may lead to prolongation of acid clearance from the oesophagus. PMID- 2979218 TI - Twenty-four-hour intragastric acidity and clinical trial of bedtime enprostil 70 micrograms compared with ranitidine 300 mg in duodenal ulcer. AB - The effects of bedtime 70 micrograms and twice daily 35 micrograms doses of enprostil on 24-hour intragastric acidity were investigated in nine duodenal ulcer patients in remission. Median nocturnal acidity decreased significantly by 30% with 35 micrograms twice daily, and by 48% with 70 micrograms at bedtime. In a clinical trial using bedtime dosing, 102 duodenal ulcer patients randomly received either ranitidine 300 mg or enprostil 70 micrograms. More ulcers healed after 4 and 8 weeks treatment with ranitidine than with enprostil (76% ranitidine vs 52% enprostil, at 4 weeks p = 0.0065 and 94% vs 68%, respectively at 8 weeks, P = 0.0007). However, 6 months after cessation of treatment there was no material difference in overall outcome. Despite combining mucosal protection with acid inhibition enprostil treatment conferred no advantage over simple acid inhibition. PMID- 2979219 TI - Review: antisecretory drugs: cellular mechanisms of action. AB - Parietal cell secretory function may be inhibited by three mechanisms. (1) Receptors for gastrin, histamine and acetylcholine are present on the canine parietal cell, and parietal cell function may be directly inhibited by specific antagonists for each of these receptors. (2) Receptor activation of parietal cell function is mediated by cyclic AMP-dependent (histamine) and calcium-dependent (cholinergic agents and gastrin) mechanisms. The antisecretory action of prostaglandins reflect interference with histamine activation of adenylate cyclase. The current generations of calcium channel blockers have only weak antisecretory actions in vivo and are unlikely to be useful in clinical practice. (3) A third mechanism of inhibition is blockade of H+/K(+)-ATPase by substituted benzimidazoles, such as omeprazole. Each of these three mechanism provides modalities of potential clinical usefulness for treating acid-peptic disease. Gastrin and acetylcholine receptors are present on other fundic cells, in addition to the parietal cell. These other cells include the somatostatin cell in the dog fundic mucosa and the histamine-containing enterochromaffin-like (ECL) cell present in the fundic mucosa of several species. The relative impact of these receptors on different cell types on the regulation of acid secretion remains uncertain, and is probably variable among different species. One gastrin receptor of considerable importance is the gastrin receptor that exerts a trophic effect on the ECL cell in the fundic mucosa. Sustained hypergastrinaemia in response to profound hypochlorhydria is associated with hyperplasia of this cell type; the elucidation of the conditions that promote this hyperplasia and the clinical consequences of this association are pressing challenges. PMID- 2979220 TI - Review: clinical trials in peptic ulcer disease--problems of methodology and interpretation. AB - This review focuses on the methodology and interpretation of drug trials in peptic ulcer disease. The problems of planning and conduct that are discussed include the ethics of using placebo, eligibility criteria, estimations of sample size, stopping rules, randomization, blinding, and efficacy criteria, that is, ulcer healing and pain relief in the short term and prevention of relapse and complications in the long term. Statistical topics covered include confidence intervals, evaluation of survival type data, post-stratification, and sub-group analysis. The difference between clinical and statistical significance is discussed, major problems being overemphasis on P-value, type II errors, and post hoc power determinations. Explanatory and pragmatic questions are based on compliance-to-protocol and intention-to-treat cohorts, respectively, and involve problems of compliance testing, evaluation of withdrawals, and the use of fixed dose regimens. The rather slow process for clinical trials to gain acceptance is described, and it is proposed to rely on disease-related and behavioural barriers, lack of knowledge of the inherent limitations of methodology, and overemphasis on the subject of peptic ulcer healing, in addition to some concern at the relevance of assessing long-term drug intervention by repeated endoscopies rather than by studying symptoms and the incidence of complications. We foresee an increased impact of clinical trials on ulcer research and therapeutic decision making, provided physicians are able to develop the proper methodology to answer the relevant questions. PMID- 2979221 TI - Review: post-marketing surveillance of the safety of cimetidine--the problems of data interpretation. AB - Experience obtained during post-marketing surveillance of the safety of cimetidine emphasizes the difficulties in interpretation posed by the high background frequency of disease of all types in drug takers. The multiple sources of confounding factors, and their high prevalence, make it impossible to detect adverse events which mimic ordinary disease, particularly when a consistent relationship between adverse event and drug exposure is not observed. The inclusion of controls emphasizes the difficulties but does not ease interpretation. PMID- 2979222 TI - Review: loperamide--a potent antidiarrhoeal drug with actions along the alimentary tract. AB - The synthetic opioid, loperamide, reduces stool weight, frequency of bowel movements, urgency and faecal incontinence in acute and chronic diarrhoea. In man, the mechanism of action of loperamide is primarily the retardation of small intestinal transit, and the stimulation of anal sphincter pressure and of faecal continence. This mechanism increases mucosal contact time, allowing more complete absorption of electrolytes and water. Studies in animals have demonstrated inhibitory effects of opiates and opioids, including loperamide, on fluid and electrolyte secretion induced by various secretagogues. By comparison, opiates have smaller if any antisecretory or pro-absorptive actions in man. The discrepancies between the results obtained in animal and human experiments are most certainly due to the large differences between drug doses used. Besides its opiate-receptor binding and stimulating activity, loperamide also behaves as a calcium-calmodulin antagonist and as a calcium channel blocker. These two other mechanisms might contribute to loperamide's antidiarrhoeal activity. Loperamide is more effective and safer than other opiates or opioid drugs in the treatment of both infantile and adult diarrhoea of various causes, although adequate fluid and electrolyte replacement remain the prime need. PMID- 2979223 TI - Gastrointestinal transit of an enteric-coated delayed-release 5-aminosalicylic acid tablet. AB - Gastrointestinal transit of an enteric-coated, delayed-release 5-aminosalicylic acid tablet radiolabelled with indium-111 has been monitored in a preliminary study with eight healthy subjects using gamma scintigraphy. Gastric emptying of the tablet was delayed by the presence of food in the stomach. Disintegration occurred about 5 hours after the tablet left the stomach. There was close agreement between the tablet disintegration times and the initial detection of drug in the blood. The site of disintegration could be established in most instances; approximately 80% of the doses resulted in drug dispersion within the ascending colon. The coated tablets provide an effective means of drug delivery to the proximal colon. PMID- 2979224 TI - Comparison of ranitidine 300 mg twice daily, 300 mg at night and placebo on 24 hour intragastric acidity of duodenal ulcer patients. AB - Continuous measurement of 24-hour intragastric acidity was performed in 12 duodenal ulcer patients in remission during treatment with placebo, ranitidine 300 mg nocte and ranitidine 300 mg b.d. Median 24-hour acidity was 79.4 mmol litre-1 during placebo treatment; it decreased to 28.2 mmol litre-1 during treatment with ranitidine 300 mg nocte and to 3.6 mmol litre-1 during treatment with ranitidine 300 mg b.d. The two regimens decreased intragastric acidity to a similar degree during the night, but significantly greater inhibition of daytime and 24-hour acidity followed use of ranitidine 300 mg b.d. PMID- 2979225 TI - Twenty-four-hour intragastric acidity and plasma gastrin concentration in healthy subjects and patients with duodenal or gastric ulcer, or pernicious anaemia. AB - Twenty-four-hour intragastric acidity and plasma gastrin concentration were measured in healthy subjects (n = 16), and patients with duodenal (n = 12) or gastric (n = 10) ulceration, or pernicious anaemia (n = 8). Median integrated 24 hour intragastric acidity was highest in duodenal ulcer patients and lowest in pernicious anaemia patients (1148 and 0 mmol.hour litre-1, respectively). Median integrated 24-hour plasma gastrin was highest in pernicious anaemia and lowest in the healthy subjects (9886 and 238 pmol.hour litre-1, respectively). Pernicious anaemia patients have unremitting hypergastrinaemia throughout the 24 hours. The results of this study not only provide a reference range of acidity and plasma gastrin in health and disease, but also will act as a baseline for future studies using antisecretory drugs. PMID- 2979226 TI - Twenty-four-hour intragastric acidity and plasma gastrin concentration before and during treatment with either ranitidine or omeprazole. AB - Simultaneous 24-hour intragastric acidity and plasma gastrin concentrations were measured in 12 duodenal ulcer patients before and on the twenty-eighth day of treatment with either ranitidine 150 mg b.d. or omeprazole 20 mg o.m. Median integrated 24-hour intragastric acidity was decreased significantly from 1148 to 490 and 36 mmol.hour litre-1 during treatment with ranitidine and omeprazole, respectively, whilst median intragastric 24-hour plasma gastrin was raised significantly from 328 to 799 and 1519 pmol.hour litre-1 respectively. When the results of all 48 experiments were considered together, there was a significant inverse correlation between the 24-hour integrated values for intragastric acidity and plasma gastrin concentration. Both drugs caused a significant elevation of plasma gastrin throughout the 24 hours, although ranitidine had no effect on intragastric acidity from 1900 to 2200 hours. When compared with similar profiles of acidity and gastrin in pernicious-anaemia patients, the modest elevations of plasma gastrin observed in this study suggest that neither drug will be associated with clinically relevant enterochromaffin-like cell proliferation in duodenal ulcer patients. PMID- 2979227 TI - Gastric acid secretory capacity falls after repeated within-day pentagastrin testing in fed subjects. AB - Between-day pentagastrin testing yields highly reproducible stimulated gastric acid output values, but little is known of the reproducibility of repeated within day pentagastrin tests. We have performed three pentagastrin tests within the 1 day in nine healthy subjects. Within-day tests were 6 hours apart; the first followed an overnight fast and the second and third were both 4 hours after a substantial meal. A further test was performed the following morning, again after an overnight fast, which allowed comparison of within-day and between-day testing. In the second and third within-day tests there was a marked decrease of stimulated gastric acid output, with both maximal and peak acid output decreased to approximately half of the value of the first test (P less than 0.01). By contrast there were no significant differences in the acid output values obtained in between-day tests (both following an overnight fast). Possible mechanisms for the decreased output on repeated within-day testing include alterations in the sensitivity of the gastrin receptor, or some neurohumoral influence secondary to the preceding meal. Future studies of the duration of action of drugs affecting acid secretion may need to take account of these findings. PMID- 2979228 TI - Review: the treatment of primary biliary cirrhosis. AB - Considerable improvement in quality of life can be achieved by treating the complications of primary biliary cirrhosis. However, once fibrosis, cirrhosis, portal hypertension and hyperbilirubinaemia occur, it is unlikely that 'medical' treatment will cure the disorder. Trials of medical therapy should aim to delay or halt the progression of the early phase of the disease and clinicians should refer patients to centres conducting clinical trials. If the disease progresses, and the quality of life is seriously impaired, liver transplantation offers the realistic chance of a new life. PMID- 2979229 TI - Omeprazole in the treatment of Zollinger-Ellison syndrome: a 4-year international study. AB - The H+, K(+)-ATPase inhibitor omeprazole has been made available on a compassionate basis for patients with Zollinger-Ellison syndrome considered resistant to, or with side-effects on, histamine H2-receptor antagonists. By December 1985, the first 80 patients, from 46 centres in 11 countries, had been treated for periods of 2 days to 4 years. Basal acid output was decreased to the desired level of less than 10 mmol.hour-1, and symptoms were rapidly relieved. Acid secretion and laboratory variables were checked regularly and endoscopic examinations made at intervals. Dosage was adjusted primarily on the basis of basal acid output, but also if symptoms recurred. The starting dose was generally 60 mg daily and the median dose ranged between 60 and 70 mg daily over the study period. There was no evidence of tachyphylaxis. More than 90% of the patients were successfully controlled on total daily doses of 120 mg or less; one-third of patients required divided doses. Tolerance was good: there were no obvious drug related effects on laboratory variables, including fasting serum gastrin, and there were very few adverse events. Thirteen patients died (11 of the primary disease and two of other causes), four underwent successful tumour resection, six underwent total gastrectomy (though acid secretion was controlled in five), four patients' treatment was changed to other medical therapy, and one was lost to follow-up. Omeprazole thus appears to be both safe and very effective in controlling acid hypersecretion in this group of patients, and to provide a suitable alternative to total gastrectomy. PMID- 2979230 TI - Double-blind comparison of 5-aminosalicylic acid and acetyl-5-aminosalicylic acid suppositories in patients with idiopathic proctitis. AB - Suppositories containing 300 mg 5-aminosalicylic acid (1.96 mmol) or 425 mg acetyl-5-aminosalicylic acid (1.96 mmol) were used in 40 patients with idiopathic proctitis to determine the efficacy of acetyl-5-aminosalicylic acid in treating this bowel inflammation. Each patient was treated with 5-aminosalicylic acid or acetyl-5-aminosalicylic acid suppositories twice daily for 4 weeks in a double blind trial. Four patients were included twice in the trial. The second time they were treated with the alternative regimen. Six patients in the acetyl-5 aminosalicylic acid group did not complete the trial, four of them because of diarrhoea. Complete clinical remission with normal rectal mucosa on sigmoidoscopy was achieved in 10 out of 18 patients on 5-aminosalicylic acid and in only two out of 15 in the acetyl-5-aminosalicylic acid group (P = 0.03). A favourable histological improvement was demonstrated with 5-aminosalicylic acid suppositories, but the difference with acetyl-5-aminosalicylic acid was not significant (P = 0.059). Three of the four patients who received both drugs recovered with 5-aminosalicylic acid; in none of them was acetyl-5-aminosalicylic acid effective. The results from this study and from previous investigations show that acetyl-5-aminosalicylic acid is not superior to placebo. PMID- 2979231 TI - Comparison of groups when outcomes are on an ordered scale. PMID- 2979232 TI - RIDIT analysis for qualitative and subjective clinical data. PMID- 2979233 TI - Pharmacological manipulation of adynamic ileus: controlled randomized double blind study of ceruletide on intestinal motor activity after elective abdominal surgery. AB - In a double-blind placebo-controlled trial of patients undergoing elective abdominal surgery (n = 91), a single intravenous infusion of ceruletide (2.5 ng kg-1 min-1 for 1 hour) resulted in audible bowel sounds in 42/47 patients as opposed to 30/44 receiving placebo (P less than 0.025). Excessive bowel sounds were noted in 16 patients in the ceruletide group and four receiving placebo (P less than 0.01). Significantly more patients (P less than 0.01) in the ceruletide group (22/45 versus 9/44) passed flatus per rectum between the second and third post-operative day. Ceruletide infusion was accompanied by a significant increase in the incidence of nausea and vomiting (P less than 0.005, P less than 0.0025) but these side effects were short-lived. These results indicate that ceruletide is likely to be a useful therapeutic agent for acute intestinal adynamic motility disorders. PMID- 2979234 TI - A comparison of two ranitidine intravenous infusion regimens in critically ill patients. AB - The effect of two ranitidine intravenous infusion regimens on intragastric pH was studied in 134 critically ill patients admitted to 15 intensive care units. Intragastric pH was determined hourly for 30 hours. Those patients whose intragastric acidity fell below pH 4.0 for 3 or more of the first 6 hours were considered 'at risk' of developing stress-related gastric lesions and randomized to receive a 50 mg bolus of ranitidine together with a continuous intravenous infusion of either 0.125 or 0.25 mg kg-1 h-1 ranitidine for 24 hours. The maximal elevation in intragastric pH was achieved within 12 hours. The median intragastric pH for the last 20 hours of the infusion period was 5.9 for the higher dose group and 5.6 for the lower dose group. The increase in intragastric pH achieved by the two dosage regimens did not differ significantly throughout the 24 hour period. Patients having two or more of five major risk factors (head injury, major trauma, sepsis, respiratory failure/insufficiency and major surgery) had better overall control of intragastric pH on the higher dose of ranitidine than those receiving the lower dose. The majority of intensive care patients are likely to receive satisfactory treatment with the lower dosage regimen that was tested (0.125 mg kg-1 h-1). Those with multiple risk factors may, however, require treatment with higher doses of ranitidine (0.25 mg kg-1 h 1). PMID- 2979236 TI - Pharmacological response to cimetidine and healing of duodenal ulceration: effects of high-dose cimetidine and combination of cimetidine with pirenzepine. AB - Overnight gastric secretion was studied in 32 patients with acute duodenal ulcers before treatment and whilst taking cimetidine 400 mg b.d. After 6 weeks of treatment with cimetidine 400 mg b.d. 13 patients had healed ulcers, seven patients had healed ulcers but a persistent erosive duodenitis, and 12 patients had persisting ulceration. Inhibition of nocturnal gastric secretion by cimetidine 400 mg b.d. was most profound in patients who healed their ulcers completely; a less profound inhibition of nocturnal gastric secretion was seen in the non-healing and duodenitis groups. In patients with persisting ulcers and poor inhibition of nocturnal gastric secretion by cimetidine, gastric secretion could be suppressed by either cimetidine 400 mg b.d. in combination with pirenzepine 50 mg b.d., or by cimetidine 1600 mg nocte, but suppression of nocturnal gastric secretion was more effective with cimetidine 1600 mg than cimetidine with pirenzepine. PMID- 2979235 TI - Effect of enprostil on basal and meal-stimulated gastric acid and pepsin secretion, serum gastrin and gastric emptying in healthy persons. AB - Ten healthy volunteers took part in a double-blind, randomized, cross-over study of the effect of single doses of enprostil (70 micrograms) and placebo on basal and meal-stimulated gastric acid, pepsin secretion and serum gastrin. Meal stimulation was induced by modified sham feeding combined with repeated gastric instillation and withdrawal of meat soup. When studied between 1 and 2.5 hours after oral administration of the drug, enprostil decreased basal acid output by 92% (P less than 0.001) and stimulated acid output by 70% (P less than 0.001). Basal and stimulated volumes of gastric juice were decreased by 50% (P less than 0.02) and 35% (P less than 0.002), respectively. Enprostil decreased stimulated pepsin output by 34% (P less than 0.05), but had no effect on the concentration of pepsin. Neither basal nor stimulated serum gastrin concentrations were affected by enprostil. Percent recovery of the meal was measured by an unabsorbable marker, polyethylene glycol, instilled into the stomach mixed with the soup. Polyethylene glycol recovery decreased from 89% with placebo to 67% with enprostil (P less than 0.01), indicating an enhanced gastric emptying rate with enprostil. PMID- 2979237 TI - Effect of oral testosterone treatment on serum concentrations of sex steroids gonadotrophins and prolactin in alcoholic cirrhotic men. Copenhagen Study Group for Liver Diseases. AB - The aim of this study was to examine the serum concentrations of sex steroids and pituitary hormones in a randomly selected group of alcoholic cirrhotic men participating in a randomized, placebo-controlled study on the efficacy of oral testosterone treatment on the liver. Before treatment, patients (n = 25) had median serum concentrations of testosterone, oestradiol, non-protein bound oestradiol, non-sex hormone binding globulin (SHBG) bound oestradiol and oestrone sulphate which did not differ significantly from those of healthy controls (n = 16), but the patients had significantly (P less than 0.01) higher median serum concentrations of oestrone, luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin. The patients were randomized to treatment with either oral micronized testosterone (200 mg t.d.s.) or placebo for a median duration of 1 year. In the placebo group (n = 8), hormone concentrations at follow-up were not significantly different from those at entry apart from a significant (P less than 0.05) increase in FSH concentrations. Median concentrations of testosterone, oestrone, and oestrone sulphate increased significantly (P less than 0.05) in the testosterone-treated group (n = 17) when compared with concentrations at entry and concentrations in the placebo group. The testosterone-treated group had significantly (P less than 0.05) higher serum concentrations of non-protein bound and non-SHBG bound oestradiol when compared with concentrations at entry, but no significant changes were observed regarding serum oestradiol and prolactin concentrations. Both LH and FSH concentrations decreased significantly (P less than 0.05) in the testosterone-treated group when compared with concentrations at entry and concentrations in the placebo group. PMID- 2979238 TI - The effect of four narcotics on cholecystokinin octapeptide stimulated gallbladder contraction. AB - In an attempt to find the ideal analgesic to treat biliary tract pain we compared the effects of saline and equianalgesic doses of morphine, pentazocine, pethidine, and butorphanol on CCK-OP stimulated gallbladder emptying in healthy volunteers. Morphine produced a profound delay in gallbladder emptying while the other narcotics produced a significant delay in comparison to saline, but less than morphine. None of the drugs affected common bile duct diameter, bilirubin, liver enzymes or amylase. We recommend avoiding the use of morphine in the treatment of biliary and pancreatic pain, but although pethidine, pentazocine and butorphanol may be potentially detrimental we could find no definite superiority of one versus the others. PMID- 2979239 TI - Effect of a slow-release formula of trimoprostil on intragastric acidity in healthy volunteers. AB - We investigated the effect of a slow-release formula of trimoprostil, a prostaglandin E2 analogue, at a dose of 3 mg b.d. on circadian intragastric acidity in nine healthy volunteers using ambulatory pH-metry in a placebo controlled study. The effect of trimoprostil was long lasting (8 hours during the night). However, it lowered gastric pH on average only by 0.4 pH units. In four of the six women severe side-effects occurred in the form of abdominal cramping, metrorrhagia, and/or diarrhoea. These disadvantages may limit the clinical use of this drug. PMID- 2979240 TI - The effects of propranolol on hepatic encephalopathy in patients with cirrhosis and portal hypertension. AB - Beta adrenoreceptor blocking drugs have been used for the prevention of haemorrhage from oesophageal varices. However, it is possible that these agents, by virtue of their effects on hepatic blood flow, may impair liver function and precipitate hepatic encephalopathy. We have therefore studied the effect of the beta blocking drug propranolol on hepatic encephalopathy in patients with cirrhosis and portal hypertension. Twenty patients were randomly assigned to receive 4 weeks treatment with propranolol or an identical-looking placebo, the former given in a dose sufficient to reduce resting pulse rate by greater than or equal to 25%. Before and after treatment patients were assessed for the severity of liver disease and the presence of encephalopathy. EEG mean cycle frequency and fasting arterial ammonia concentrations were also measured, and in order to detect latent hepatic encephalopathy, each patient underwent a battery of psychometric tests. Patients were blinded as to their treatment, as were those assessing their responses. Neither propranolol nor placebo had any significant effect on the parameters measured. On propranolol median EEG mean cycle frequency fell from 9.08 ct s-1 (range 8.63-11.0 ct s-1) to 8.73 ct s-1 (range 8.27-11.44 ct s-1), and median fasting arterial ammonia concentration fell from 66 mumol litre-1 (range 40-329 mumol litre-1) to 49 mumol litre-1 (range 37-188 mumol litre-1). Psychometric test values, while initially abnormal and suggestive of latent hepatic encephalopathy in the majority of patients, did not change significantly during the study. PMID- 2979241 TI - A comparison of the effects of treatment with either famotidine 40 mg or cimetidine 800 mg nocte on gastric acid secretion and serum gastrin. AB - The effects on gastric acid secretion and serum gastrin of 4 weeks treatment with famotidine 40 mg nocte or cimetidine 800 mg nocte (at 2200 h) were studied in 16 patients with previous duodenal ulcer. Patients were studied before commencing therapy, on days 5, 12 and 26 of treatment, and on days 3, 10 and 24 after completion of therapy. In contrast to cimetidine, basal and pentagastrin stimulated gastric acid secretion measured 12 h after dosing was significantly inhibited during treatment with famotidine. In addition, with famotidine there was inhibition of stimulated gastric acid secretion at 3 and 24 days after completion of treatment. Fasting serum gastrin measured 12 h after dosing was not significantly altered by either drug. PMID- 2979242 TI - The effects of the 5-hydroxytryptamine (5HT3) receptor antagonist ICS 205-930 in the carcinoid syndrome. AB - Therapy for diarrhoea associated with the carcinoid syndrome is often unsatisfactory. In an open study ICS 205-930 (Sandoz Limited), a novel 5HT3 antagonist, controlled diarrhoea in five of six patients studied. This drug may be a useful advance in the symptomatic treatment of the carcinoid syndrome. PMID- 2979243 TI - The effect of nicotine on the delay of gastric emptying. AB - The effect of cigarette smoking and its active component, nicotine, on the gastric emptying of solid food was assessed in a randomized double-blind crossover design. Ten regular smokers were studied after a 6 h fast and least 18 h after their last cigarette. Subjects smoked a total of three high (1.91 mg) or low (0.17 mg) nicotine cigarettes, before and after a technetium-labelled solid meal and were scanned by gamma camera periodically over a 2-h period. All calculations of gastric emptying revealed a significant delay after smoking high versus low nicotine cigarettes in: mean per cent isotope remaining in the stomach at each measurement point from 90-120 min; amount of meal remaining in the stomach at 2 h; and mean time at which 50% of the meal had emptied (T1/2). Delay in gastric emptying was significantly correlated with increase in serum nicotine concentration on the high nicotine day. PMID- 2979244 TI - The effect of dietary fatty acids on the gastric production of prostaglandins and aspirin-induced injury. AB - We have investigated in man the effect of dietary supplementation with evening primrose oil (linoleic acid 72%, gamma-linolenic acid 9%) on gastric immunoreactive PGE2 release and aspirin-induced mucosal injury. Twenty healthy volunteers received in a randomized crossover evening primrose oil (2 g b.d.) and olive oil (control) for 2 weeks each. During the last 48 hours of each period, five doses of aspirin 900 mg were taken. In gastric washings on day 7, evening primrose oil had enhanced immunoreactive PGE2 release from a mean of 38 ng/30 min (olive oil) to 80 ng/30 min (P less than 0.05). This, however, was not able to protect against aspirin-induced gastric blood loss which rose from a mean (95% confidence limits) of 1.3 (0.7-2.1) microliters 10 min-1 (evening primrose oil day 7) to 9.4 (5.4-16.5) microliters 10 min-1 (day 14). Although not effective against aspirin, similar dietary supplementation might assist in preventing peptic ulcer recurrence. PMID- 2979245 TI - The relationship between plasma cortisol and gastric mucosa prostaglandin levels in rats with stress ulcers. AB - A correlation between the levels of plasma cortisol, gastric mucosal prostaglandins and the degree of gastric ulceration produced by stress in rats was examined. Four groups of rats were evaluated. Group A, the cold stressed group; group B, their controls; group C, those receiving intraperitoneal indomethacin; and group D, those receiving intraperitoneal saline. Group A developed stress ulcers and their gastric mucosal prostaglandin levels (prostaglandin E2 (PGE2) and prostacyclin levels, as measured by its stable metabolite 6-keto-prostaglandin F1 alpha (6-keto-PGF-1 alpha, but not thromboxane) were significantly reduced when compared with their respective non stressed controls. The plasma cortisol levels in both group A and B increased slightly in the first hour but there was no statistical difference between the two groups and there was no change at 2, 3 or 4 h of stress. Group C (indomethacin) also developed ulcers and had low gastric mucosal prostaglandin levels when compared with group D (saline). The plasma cortisol levels did not alter in either group C or D. It has been postulated that stress ulcers may involve the depletion of gastric mucosal prostaglandin levels which, in turn, may be a consequence of a higher plasma cortisol level. A decrease in gastric prostaglandins independent of any change of plasma cortisol levels was demonstrated in this study and the mechanism of production of stress gastric ulcers remains obscure. PMID- 2979246 TI - The effects of omeprazole and ranitidine on 24-hour pH in the distal oesophagus of patients with reflux oesophagitis. AB - Ambulatory 24-h pH monitoring in the distal oesophagus was performed in seven patients with erosive or ulcerative reflux oesophagitis to compare the effects of omeprazole and ranitidine in the management of gastro-oesophageal reflux disease. In a double-blind, crossover study patients were treated with either 60 mg o.m. omeprazole or 150 mg b.d. ranitidine. The pH measurements were performed before treatment and on the fourteenth day of treatment with either regimen. The total acid exposure time (percentage total time pH less than 4) was abnormal in six out of seven patients before treatment. During treatment with omeprazole the acid exposure time of five patients was normal in comparison with only two patients during ranitidine therapy. However, even with a rather high dose of omeprazole, pathological gastro-oesophageal reflux may still occur. PMID- 2979247 TI - Dose--response effect of famotidine on patterns of gastro-oesophageal reflux. AB - The present study attempts to assess the alteration in patterns of gastro oesophageal reflux as assessed by 24-h oesophageal pH monitoring by varying degrees of H2-receptor blockade with famotidine. Subjects were 12 patients with complaints of daily heartburn who demonstrated at least 6% of acid mucosal contact time by 24-h ambulatory oesophageal pH monitoring. All subjects had a positive Bernstein test, and nine of the 12 subjects had erosive oesophagitis. The study was conducted as a double-blind crossover design utilizing 40 mg nocte, 20 mg b.d., and 40 mg b.d. and placebo treatments. Results indicated that all treatments significantly reduced the 24-h percentage acid contact time (P less than 0.05) compared to placebo. The two b.d. treatment regimens also significantly (P less than 0.05) reduced the number of episodes lasting longer than 5 min. Only the b.d. regimens successfully lowered the percentage of upright acid exposure. All treatments significantly (P less than 0.01) reduced the percentage of supine acid contact time, as well as the number of episodes lasting more than 5 min. It is concluded that gastro-oesophageal reflux disease may well require a b.d. dosing regimen with famotidine in order to achieve optimal mucosal healing and day time symptom control. PMID- 2979248 TI - Prophylaxis of aspirin-induced gastric mucosal bleeding with ranitidine. AB - The ability of ranitidine to protect the human gastric mucosa against aspirin induced damage was investigated by timed measurements of blood loss collected by gastric washing. Ranitidine (150 mg) 1 h or 5 h before 900 mg aspirin (5 doses of each) over 48 h reduced subsequent mean bleeding from 7.7 microliters/10 min to 2.6 microliters/10 min or 3.4 microliters/10 min, respectively. Both regimens were antisecretory at the time of aspirin administration, as judged by a rise in the pH of the aspirated washings. The prolonged protection against aspirin induced bleeding achieved with twice daily dosing with ranitidine has clinical potential in the management of patients taking anti-inflammatory drugs. PMID- 2979249 TI - Oral rehydration therapy without bicarbonate for prevention and treatment of dehydration: a double-blind controlled trial. AB - Forty children (less than or equal to 2 years of age) were admitted to hospital with acute gastroenteritis and were randomly assigned to receive either an oral rehydration solution (ORS) containing bicarbonate (Na 35, K 20, Cl 37, HCO3 18, glucose 202 mmol litre-1) or an identical solution in which bicarbonate was replaced by chloride ions. Groups were matched for age, sex, ethnic origin, duration of diarrhoea and nutritional status. On admission, degree of dehydration, biochemical and haematological parameters were similar. The majority had minimal or no dehydration and only 30% had moderate to severe dehydration. All children were treated successfully with no complications. Oral rehydration solution intake by each group was similar. Clinical outcome, as judged by speed of rehydration or maintenance of hydration, duration of diarrhoea, stool frequency and length of hospital stay, was the same in both groups. After 24 h of ORS there was no difference between groups in venous pH, serum bicarbonate, urea and electrolytes. In hospitalized children with acute gastroenteritis in the United Kingdom an ORS without bicarbonate is a safe, effective means to prevent dehydration and maintain hydration and acid-base status where dehydration is not severe. Exclusion of bicarbonate would simplify production, increase stability and reduce the cost of ORS without apparent impairment of efficacy. PMID- 2979250 TI - Effect of low-dose omeprazole on gastric acid secretion in duodenal ulcer patients. AB - The effect of 7 days of oral dosing with 5 mg day-1 and 20 mg day-1 omeprazole on basal and pentagastrin-stimulated gastric acid output was studied in nine duodenal ulcer patients. Basal acid output measured 5-6 h post-dosing was decreased by a mean of 75% on 5 mg omeprazole and by 90% on 20 mg omeprazole (P less than 0.05 and P less than 0.01, respectively). Peak acid output measured 6-7 h post-dosing was decreased by a mean of 75% on 5 mg omeprazole and 97% on 20 mg omeprazole (P less than 0.01 for each). There was a wide interindividual variation in response to the 5 mg dose, with five of the nine patients having more than 90% inhibition of peak acid output, but two patients having less than 40% inhibition. This unpredictable response to daily low-dose omeprazole therapy makes it unsuitable for maintenance treatment. PMID- 2979251 TI - Review: new aminosalicylic acid derivatives for the treatment of inflammatory bowel disease. AB - Up to 20-30% of patients treated with sulphasalazine experience a variety of adverse effects, principally due to the carrier moiety sulphapyridine. In the last decade there has been a major drive to develop a new generation of 5 aminosalicylic acid (5-ASA) and 5-ASA-related drugs which not only have a high efficacy but are also devoid of the unwanted side-effects of sulphapyridine. Various forms of 5-ASA have been evaluated in ulcerative colitis and appear to be effective orally in preventing relapse and topically in the treatment of active distal colitis. More recently, topical 4-ASA has been found to be useful for the treatment of distal colitis with the advantage of better stability and lower cost compared with 5-ASA. In the foreseeable future it seems likely that these new aminosalicylic acid derivatives will become the drugs of choice in the treatment of inflammatory bowel disease and largely replace sulphasalazine. PMID- 2979252 TI - Review: the enterochromaffin-like cell, intragastric acidity and the trophic effect of plasma gastrin. AB - Although enterochromaffin-like (ECL) cells form the major endocrine cell population of the non-antral stomach, they have been largely overlooked in the study of gastric disease. In the human, their product and functions are unknown, but they are associated with histamine secretion in rodents. The cells are controlled by neural and hormonal factors, the most significant of the latter being gastrin. Interest in ECL cells has been stimulated by the observation that hyperplasia of these cells, sometimes leading to formation of gastric carcinoid tumours, occurs in conditions of persistent hypergastrinaemia - for example, in response to the achlorhydria of individuals with pernicious anaemia. The advent of new highly potent inhibitors of gastric acid secretion is allowing more information to be obtained on the physiology and functions of the ECL cell. However, there is clearly a great deal more to be discovered about this enigmatic endocrine cell type. PMID- 2979254 TI - Enprostil inhibits post-prandial gastrin release: a dose-response study. AB - In a double-blind placebo-controlled study in nine healthy volunteers, the effects of single doses of oral enprostil (8.75, 17.5, 35 and 70 micrograms), taken before a standard breakfast, were assessed on the post-prandial release of gastrin into the plasma. All doses of enprostil caused a significant dose-related decrease in median post-prandial plasma gastrin concentration (range from -29 to 44%). In the same subjects, two doses of 25 mg indomethacin caused a significant (38%) increase in median post-prandial plasma gastrin concentration. PMID- 2979253 TI - Involvement of M1 cholinergic receptors and enteric nerves in the spasmogenic activity of bacterial N-formyl oligopeptides on guinea-pig ileum. AB - Bacterial N-formyl-methionyl oligopeptides are spasmogenic for guinea-pig ileum in vitro but the mechanism of this effect is not understood. To investigate this phenomenon further, we have determined pA2 values (the negative logarithm of the concentration of an antagonist reducing a double-dose agonist response to a single-dose response) for a number of potential antagonists of N-formyl-met-leu phe (F-met-leu-phe) using histamine, acetylcholine, 5HT and substance P as control agonists. Atropine, pirenzepine and tetrodotoxin were potent inhibitors of F-met-leu-phe induced contraction (pA2's 8.4, 8.0 and 7.9, respectively) suggesting involvement of neural and cholinergic pathways in the response. Sulphasalazine, known to block the F-met-leu-phe receptor on neutrophil leucocytes, was also a potent inhibitor. Tachyphylaxis induced by either 5HT, or substance P, did not diminish the response to F-met-leu-phe, suggesting that these potential mediators were not involved. These studies indicate that bacterially synthesized formyl-methionyl oligopeptides bind to cells bearing receptors in guinea-pig ileum and produce muscle contraction via enteric cholinergic (M1) neural pathways. PMID- 2979256 TI - Gastro-oesophageal reflux associated with nifedipine. AB - A 57-year-old woman presented with a 3-week history of dysphagia for solids, 6 months after starting treatment with nifedipine. Endoscopy demonstrated oesophagitis and a benign oesophageal stricture. Twenty-four-hour ambulatory pH monitoring demonstrated decreased acid reflux 8 weeks after withdrawal of nifedipine, with coincidental symptomatic and endoscopic improvement. Nifedipine may induce, or aggravate, pre-existing, gastro-oesophageal reflux. PMID- 2979255 TI - Twenty-four-hour intragastric measurements in twenty healthy subjects: effect of enisoprost, a novel and potent antisecretory and antipeptic synthetic E1 prostaglandin. AB - Gastric aspiration was performed continuously overnight and at hourly intervals during the daytime in 20 healthy male volunteers. Medications used included enisoprost 100, 200 or 400 micrograms, misoprostol 200 micrograms and placebo, given at bedtime. Each dose of enisoprost markedly inhibited nocturnal mean acid output, hydrogen ion activity, pH and peptic activity. The duration of these effects was up to 10 h. Misoprostol, given at bedtime, also decreased acid secretion, but the effect was significantly less than that observed with any of the doses of enisoprost. A dose-response effect for enisoprost was found for the mean nocturnal hydrogen ion activity and pH, as well as for maximum pH attained. Although enisoprost, given at bedtime, had a marked inhibitory effect on acid and pepsin secretion for the overnight interval, this did not result in rebound hyperacidity or a rise in serum total gastrin concentration. The results of this study suggest that enisoprost should be tested by clinical trial for the treatment of peptic ulcer disease. PMID- 2979257 TI - Theophylline disposition--effects of cimetidine, mebendazole and albendazole. AB - On the basis of the report that benzimidazoles bind to and inhibit the hepatic cytochrome P-450 enzyme system, the effect of mebendazole and albendazole on theophylline disposition was studied in 12 volunteers. Mebendazole at a dose of 100 mg b.d. for 3 days did not significantly alter the theophylline half-life, volume of distribution or clearance in a group of six. In another group of six adult volunteers, albendazole (400 mg) pretreatment did not alter the same parameters. However, in this second group, pretreatment with cimetidine (400 mg t.d.s. for 5 days) significantly increased theophylline half life from 7.7 to 9.8 +/- 1.5 h (P less than 0.001) and reduced its clearance from 0.8 to 0.60 +/- 0.1 ml min-1 kg-1 (P less than 0.005). The volume of distribution was not altered significantly. It is concluded that at therapeutic doses it is unlikely that mebendazole or albendazole will induce theophylline toxicity if co-administered with the bronchodilator. Cimetidine-induced impairment of theophylline metabolism is such that toxicity will be more likely in individuals with initial high theophylline clearance. PMID- 2979258 TI - A double-blind crossover comparison of lidamidine, loperamide and placebo for the control of chronic diarrhoea. AB - Fourteen patients with chronic diarrhoea, but no evidence of active organic disease, completed a double-blind crossover comparison of the anti-diarrhoeal effects of loperamide, placebo and the clonidine analogue, lidamidine. Failure of diarrhoea control led to early withdrawals from seven placebo- and six lidamidine treatment periods, but there was only one early withdrawal during treatment with loperamide. Loperamide was found to be superior to lidamidine or placebo for the control of stool consistency in patients with chronic diarrhoea. PMID- 2979259 TI - Enprostil, a prostaglandin E2 analogue, in the treatment of duodenal ulcer; a multicentre comparison with cimetidine. AB - In a multicentre trial, 120 patients with endoscopically diagnosed duodenal ulcer were randomly allocated to treatment with either 35 micrograms enprostil b.d. or 400 mg cimetidine b.d. for up to 6 weeks on a double-blind basis. After 6 weeks, 82% (42/51) of enprostil-treated patients and 92% (44/48) of cimetidine-treated patients were healed. Corresponding healing figures on an intention-to-treat basis were 70% and 76%. No significant differences were detected between treatments with respect to healing rates or symptom control at any time. Side effects were reported by 14 patients taking enprostil and 17 patients taking cimetidine; none were serious but they resulted in withdrawal of one and two patients respectively. Enprostil was found to be similar in efficacy and tolerance to cimetidine. PMID- 2979260 TI - Haemodynamic responses to 1.25 and 2 mg of terlipressin intravenously in man. AB - The systemic and hepatic haemodynamic effects following terlipressin (Glypressin) administration were studied. Bolus intravenous injections of 1.25 and 2 mg significantly decreased wedged hepatic vein pressure gradient by 29% and 31%, respectively. With each dose there was a small increase in mean arterial blood pressure but after 1.25 mg of terlipressin, estimated hepatic blood flow fell by only 11% (not significant) compared to 24% after 2 mg, while the cardiac index fell by 12% compared to 29%. The adverse haemodynamic effects of terlipressin were diminished by a decrease in dosage, without any alteration in the beneficial effect of the drug upon portal pressure. Whether the smaller dose will be effective in arresting variceal haemorrhage remains to be determined. PMID- 2979261 TI - No correlation between indomethacin-induced gastroduodenal damage and inhibition of gastric prostanoid synthesis. AB - The effect of 1 week of treatment with indomethacin 150 mg/day on human gastric prostanoid synthesis was correlated with its effect on gastric and duodenal mucosa. Before and following 1 week of treatment, endoscopic appearance of the mucosa was evaluated and scored. Following 1 week of treatment with indomethacin, antral PGE2 and 6-keto-PGF1 alpha were significantly lower than in normal subjects, but similar in patients with significant or with no mucosal damage. Co treatment with ranitidine 150 mg b.d. or with cimetidine 400 mg b.d. reduced the mean mucosal damage score but did not affect gastric prostanoid synthesis, which was similar irrespective of the presence or absence of mucosal damage. It is therefore suggested that there is no correlation between indomethacin-induced inhibition of gastric prostanoid synthesis and its induction of mucosal damage. PMID- 2979262 TI - Review: the pharmacological control of variceal bleeding. AB - The haemodynamic effects and clinical uses of drugs used in the management of patients with oesophageal variceal bleeding are reviewed. Vasoconstrictor agents (vasopressin, teripressin) alone or in combination with nitrates continue to be used for acute bleeding episodes, while somatostatin is an alternative. Alpha- and beta-adrenergic blocking drugs and vasodilators which lead to a sustained decrease in portal pressure can be used for the prevention of bleeding episodes, but despite numerous studies the pharmacological treatment of variceal bleeding remains controversial. PMID- 2979263 TI - Six years of continuous cimetidine treatment in peptic ulcer disease: efficacy and safety. Anglo-Irish Cimetidine Long-Term Study Group. AB - The management of peptic ulcer disease with up to 6 years of continuous cimetidine treatment has been studied at four centres in the UK and Ireland. Cimetidine, 1 g daily for up to 12 weeks, was used to heal duodenal or gastric ulceration, alternating with 400 mg each night to maintain remission. After an initial healing phase, 402 patients started maintenance therapy. Classical life table analyses showed that after 5 years one-quarter of the patients had suffered a symptomatic relapse, about half during the first year of maintenance treatment. Prevalence analyses (reflecting the cyclical nature of H2-antagonist use in peptic ulcer disease) over 6 years show that, on average, over 97% remain free of symptomatic ulceration at any one time. Patients 'lost' to the study due to withdrawal or default may, however, bias the results. A model designed to account for these estimates that during the long-term management of peptic ulcer with cimetidine, on average around 95% of patients remain free of symptomatic ulceration at any one time. Safety studies, including haematological and biochemical testing over the whole period, revealed no previously unknown adverse reaction. Seven patients were withdrawn for events considered to be related to the drug. Eleven patients died; none of the deaths appears to be connected with the treatment. It is concluded that long-term use of cimetidine is effective in the management of peptic ulcer disease, and continuous treatment for up to 6 years has not revealed any previously unknown hazard. PMID- 2979264 TI - Three computer models for the calculation of prevalence of peptic ulcer disease during long-term treatment. AB - At present the effects of maintenance treatment for peptic ulcer disease are usually calculated by using 'life-table' analyses. Whilst these accurately demonstrate the speed with which an initial relapse occurs they make no allowance for the fact that, in clinical practice, a relapse often responds to a further course of full-dose treatment and the patient then returns to maintenance therapy. A further compounding factor is that, in any long-term study, patients will be lost to follow-up for a variety of reasons not all related to failure of the treatment. In this paper we describe the use of 'prevalence rates' to better reflect the outcome of peptic ulcer management. Three 'computer models', which have been developed to address the problems of patients leaving the study for any reason during such a long time-period, are also described, as are the underlying assumptions made. Using the results from a long-term study of continuous treatment with cimetidine, the 'prevalence rates' of ulcer disease over 6 years were calculated. Observed relapse rates appeared to fall with time (from 2.7% for duodenal ulcer (DU) and 2.5% for gastric ulcer (GU) to 1% and 2% respectively). However, on applying the models to the data, prevalence rates tended to rise slowly with time for the first 3 years in each of the models tested. At 6 years, two of the models suggested that the prevalence rate for DU would be about 8%; this is not very different to the reported recurrence rate after surgical treatment by truncal vagotomy and pyloroplasty. It is concluded that 'prevalence rates' should be used to assess long-term medical treatments for ulcer disease. Similar methods could also be used to examine the medical treatment of any other disease where multiple relapses, capable of responding to re-treatment, occur. The use of models proved beneficial in compensating for patients lost during the study. PMID- 2979265 TI - The anti-secretory effect and pharmacokinetics of omeprazole in chronic liver disease. AB - The anti-secretory effects and pharmacokinetics of omeprazole were investigated in ten patients with chronic liver disease. Plasma omeprazole concentrations were measured after a 10-mg intravenous dose of omeprazole and on the first and seventh days of a 7-day course of 10 mg oral omeprazole daily. Pentagastrin tests were performed on the day before oral omeprazole was commenced and 24 h after the last oral dose. The pre-treatment basal and peak gastric acid outputs were low (mean rates of 1.44 mmol/h and 9.26 mmol/h, respectively) and following 7 days of oral 10 mg omeprazole daily, were lowered by 95% and 90% respectively. Following 10 mg intravenous omeprazole, plasma clearance was reduced, and plasma half-life and area under the concentration curve were increased, in comparison with previous studies in healthy subjects. The plasma concentration curves for oral and intravenous doses were very similar. After both the first and seventh oral doses, maximum plasma concentration and area under the curve were higher than in healthy subjects. No accumulation of omeprazole was demonstrated. The pharmacokinetics of omeprazole in chronic liver disease could be influenced by low gastric acidity, poor liver function and/or portasystemic shunting. A dose of 10 mg omeprazole daily has been shown to be an effective anti-secretory agent in chronic liver disease. PMID- 2979266 TI - Comparative irritancy of oxaprozin on the gastrointestinal tract of rats and mice: relationship to drug uptake and effects in vivo on eicosanoid metabolism. AB - Studies were performed in rats and mice to investigate the mechanisms of the comparatively low irritancy in the gastrointestinal (GI) tract of oxaprozin, relative to that of other non-steroidal anti-inflammatory drugs. A single dose of oxaprozin (100 mg/kg), given orally to rats, reduced the concentration of prostaglandin E2, but did not produce any gastric lesions or mucosal irritation, either visibly or when the mucosa was examined by scanning or transmission electronmicroscopy. The low gastric irritancy was reflected by lower concentrations of the drug in the gastric mucosa, compared with those in the upper intestinal tract, following oral administration of 100 mg/kg of [14C]oxaprozin. This distribution of the drug in the gastrointestinal tract reflected the absence of gastric irritancy when the drug was given repeatedly for 5 days by the oral route to rats. It was only after intraperitoneal administration of high doses of the drug that intestinal ulcers and peritonitis became evident, and then no gastric irritancy was observed. As with other non steroidal anti-inflammatory drugs, there was some, but appreciably less, enhancement of gastric irritancy observed when the drug was given to mice treated with the cholinomimetic, bethanechol chloride (to enhance secretion of acid and pepsin). It is suggested that the inhibition of mucosal prostaglandin production by oxaprozin may be considered a 'priming' reaction, and that subsequent enhancement of irritancy occurs by stimulation of acid and pepsin. Structure- activity relationships of oxaprozin analogues were studied. The addition of electron-withdrawing substituents, which markedly enhance the gastric ulcerogenic effects of other non-steroidal anti-inflammatory drugs, only slightly enhanced the irritancy of oxaprozin, thus providing further evidence for the inherent low ulcerogenicity of this molecule. PMID- 2979267 TI - The effect of a long-acting somatostatin analogue on portal and systemic haemodynamics in cirrhosis. AB - Current interest in the pharmacological manipulation of portal pressure centres on the long-acting somatostatin analogue SMS 201-995. Nine haemodynamically stable cirrhotic patients who had previously bled from oesophageal varices had wedged and free hepatic venous pressures and cardiac index measured, using a Swan Ganz catheter, before and at 60, 120 and 180 min after beginning a 60-min infusion of 25 microgram/h of SMS 201-995. Seven clinically similar patients had the same measurements performed without SMS 201-995. In all patients cardiac index was found to decrease and systemic vascular resistance increase at 60 min, although heart rates and arterial blood pressures were unchanged. The group given SMS 201-995 was significantly different from the control group in sustaining a fall in wedged hepatic venous pressure and trans-hepatic venous gradient at 60 min. SMS 201-995 causes a fall in portal pressure without a significant systemic haemodynamic effect. PMID- 2979268 TI - A model of an 'artificial stomach' for assessing the characteristics of an antacid. AB - A model of an 'artificial stomach' has been constructed in order to take into account some of the parameters lacking in conventional in vitro antacid evaluation, namely the interaction between secretory flux and variation in emptying fluxes, the presence of proteins, and the use of human gastric juice instead of an aqueous solution of hydrochloric acid. The 'artificial stomach' has two elements, the 'stomach' and the pH recording system. The 'stomach' includes a 'gastric' reservoir receiving secretory flux and is emptied by variable fluxes. Aluminium phosphate gel has been studied in 100 ml of 0.1 N HCl, without and with 1 or 5% meat extracts and also in 100 ml of human gastric juice. The antacid effect of 1 or 5% meat extracts has also been assessed. The antacid effect of aluminium phosphate was characterized by the pH rise of the 'gastric' contents, the buffering capacity, and the dilution of gastric contents. These factors were modulated by emptying fluxes. The same characteristics were found when antacid was studied in gastric juice. Proteins exerted a neutralizing effect and modified aluminium phosphate's antacid capacity. A mechanism for buffering capacity by cation aluminium is suggested. PMID- 2979269 TI - Treatment of steatorrhoea in cystic fibrosis: a comparison of enteric-coated microspheres of pancreatin versus non-enteric-coated pancreatin and adjuvant cimetidine. AB - Enteric-coated microspheres of pancreatin were compared with non-enteric-coated pancreatin combined with cimetidine taken 40 min before meals in the treatment of patients with cystic fibrosis. Fourteen adults with steatorrhoea due to cystic fibrosis were investigated in an open, randomized crossover study, over two consecutive 28-day treatment periods. Lipase intake was adjusted to each patient's previous requirements and was the same during both months; they were instructed to continue with their normal diet. Patients collected faeces for 72 h at the end of each month and completed diary cards daily throughout. Bowel actions were less frequent on enteric-coated microspheres of pancreatin than on non-enteric-coated pancreatin/cimetidine (1.7 vs. 2.4/day; P less than 0.001) and stool character was improved (P less than 0.001). Mean daily faecal weight was similar on enteric-coated microspheres of pancreatin to that on the combination (254 g vs. 291 g; N.S.), whereas daily faecal fat excretion tended to be less on enteric-coated microspheres of pancreatin (21 g vs. 27 g; N.S.), and percentage fat absorption tended to be greater (81% vs. 73%; N.S.). Mean body weight increased by 0.3 kg on enteric-coated microspheres of pancreatin and fell by 0.1 kg on the combination (N.S.). These data indicate that enteric-coated microspheres of pancreatin are at least as effective as non-enteric-coated pancreatin with cimetidine in the treatment of steatorrhoea in cystic fibrosis. PMID- 2979270 TI - The effects of omeprazole and cimetidine on duodenal ulcer healing and the relief of symptoms. AB - In a Swedish double-blind multicentre study, omeprazole (30 mg o.m.) was compared with the H2-receptor antagonist cimetidine (400 mg b.d.) in 152 patients. Clinical assessments and laboratory investigations were carried out at 2 and 4 weeks, and again at 6 weeks in unhealed patients. Endoscopy was performed at 2 weeks, and again at 4 and 6 weeks in unhealed patients. The patients in the two groups were well-matched prior to treatment. Omeprazole was superior to cimetidine in ulcer-healing rate after 2, 4 and 6 weeks. After 2 weeks of treatment, 66% of the omeprazole- and 45% of the cimetidine-treated patients were healed (P = 0.02), after 4 weeks 97 and 84% (P = 0.01), and after 6 weeks 100 and 92% (P = 0.02), respectively. There was a more pronounced improvement in the patients' symptoms in the omeprazole group after 2 weeks (P = 0.05). Both drugs were well-tolerated, but there was a high prevalence of patients with adverse events in the cimetidine group (51%, compared to 30% of the omeprazole group; P = 0.02). A total of 125 patients were followed for 6 months after healing. The patients were investigated by endoscopy after 6 months, or whenever symptoms occurred. There was no significant difference in the rate of relapse within 6 months between the two treatment groups: 54% relapsed in the omeprazole group and 52% in the cimetidine group. In conclusion, 30 mg of omeprazole, given once daily, is superior to 400 mg of cimetidine twice daily in duodenal ulcer healing; but ulcer relapse in the two groups appears to be equivalent. PMID- 2979271 TI - The benefit of pancreatic enzyme substitution after total gastrectomy. AB - The aim of the present study was to evaluate the effect of the pancreatic enzyme preparation Kreon on abdominal symptoms, bowel habits, faecal fat excretion and oro-caecal transit time in patients after total gastrectomy for carcinoma of the stomach with Roux-en-Y anastomosis. A hydrogen breath test was carried out in each patient to detect bacterial overgrowth. In a double-blind crossover trial, 15 patients were treated with either Kreon or placebo (3.6 g/day) in two test periods each of seven days. During treatment with the active substance, the stool consistency became more solid (P less than 0.05). The number of bowel movements and the abdominal symptoms, however, remained statistically unchanged. Treatment with Kreon did not influence the oro-caecal transit time. Faecal fat excretion did not significantly decrease in the whole group of patients. However, in those patients with massive steatorrhoea (free and esterified fatty acids greater than 350 mmol/72 h; upper reference limit 60 mmol/72 h) a significant (P less than 0.05) reduction from a median excretion of 643 mmol/72 h to 501 mmol/72 h was seen. Such a decrease in faecal fat excretion did not occur in patients with steatorrhoea below this limit. Bacterial overgrowth or rapid upper intestinal transit were not over-represented in patients with a high-degree of steatorrhoea. We conclude that after total gastrectomy pancreatic enzyme substitution reduces massive steatorrhoea and improves stool consistency. PMID- 2979272 TI - A double-blind placebo-controlled trial of BRL 24924 on lower oesophageal sphincter pressure and gastro-oesophageal reflux in healthy volunteers. AB - BRL 24924 is a new gastrointestinal prokinetic agent with properties similar to metoclopramide but with increased potency and devoid of side-effects associated with blockade of dopamine receptors in the central nervous system. A double-blind placebo-controlled trial of the effect of a single oral dose of 2.2 mg BRL 24924 on lower oesophageal sphincter pressure and gastro-oesophageal reflux has been performed in 20 healthy volunteers. BRL 24924 significantly increased mean lower oesophageal sphincter pressure (21.9 cmH2O BRL; 15.9 cmH2O placebo: P less than 0.017) but failed to alter either the frequency or the duration of gastro oesophageal reflux after provocation following a test meal. BRL 24924 has significant effects on lower oesophageal sphincter pressure but no effect on provoked post-prandial reflux in healthy volunteers. Further studies in patients with gastro-oesophageal reflux and oesophagitis are needed to evaluate the clinical efficacy of this compound. PMID- 2979273 TI - The effect of food on ranitidine-induced inhibition of nocturnal gastric secretion. AB - Overnight gastric secretion of acid and pepsin was studied in nine, healthy, male volunteers on two separate occasions, in randomized order. On one day, each individual received a standard meal at 18.00 hours, followed by an oral dose of 300 mg ranitidine at 18.15 hours. During the second study, 300 mg ranitidine was again taken at 18.15 hours, but the meal was consumed at 22.00 hours. Gastric juice was sampled hourly from 19.00 to 24.00 hours and then aspirated completely and continuously until 08.00 hours the next morning. Median pH values were significantly lower and median acidity was significantly higher, when the meal was eaten at 22.00 hours. The outputs of acid and pepsin after midnight were also significantly greater following the late night meal. It seems that food interferes with the gastric inhibitory effects of the currently used H2-receptor antagonists, such as ranitidine. We conclude that H2-receptor antagonists must be taken after the last meal of the day in order to ensure maximal therapeutic efficacy. PMID- 2979274 TI - A comparative study of the effects on colon function caused by feeding ispaghula husk and polydextrose. AB - Polydextrose is a new soluble food ingredient which cannot be digested by intestinal enzymes and so may affect colonic function. Studies in healthy volunteers compared the effects of diet supplementation with 30 g/day polydextrose, a standard dose of 7 g/day ispaghula and two mixtures containing 2 g/day ispaghula with either 30 g/day polydextrose or 10 g/day polydextrose with a control period. During the 10-day periods, the mass, frequency and consistency of faeces were assessed as well as the whole-gut transit time, ease of defaecation, flatulence and palatability of the preparations. All preparations significantly increased the weekly faecal mass above control values (P less than 0.05) but there were no significant differences between the preparations. Transit time and stool frequency were not affected significantly by any of the preparations (P greater than 0.05). Both preparations supplying 30 g/day polydextrose softened stool consistency equally but the other preparations had no effect. All preparations caused flatulence and other gas-related problems but polydextrose caused more than ispaghula, even at the lowest dose of 10 g/day. More volunteers preferred taking the polydextrose drinks than the sachets of ispaghula which formed a viscous drink with water. Despite superior palatability and equally effective stool bulking, polydextrose is unlikely to be an alternative laxative to ispaghula because of the unacceptable levels of flatulence. PMID- 2979275 TI - The effects of ranitidine and cimetidine on the pharmacokinetics and pharmacodynamics of metoprolol. AB - The impact of cimetidine, ranitidine and placebo on the pharmacokinetics of metoprolol, given either as a single dose (100 mg) or for 7 days (100 mg b.d.), has been evaluated in two separate studies. The doses used were 800 mg cimetidine daily and 300 mg ranitidine daily. The subjects were all young, healthy volunteers. In the single dose study, cimetidine produced a marked increase in the peak plasma concentration of metoprolol and in the area under the plasma concentration-time curve; ranitidine had less effect, though the area under the curve was significantly greater than placebo. In the chronic dosing study, the area under the curve for metoprolol was also significantly higher on cimetidine (1796 ng h/ml; P less than 0.001) whereas the area under the curve on ranitidine (1258 ng h/ml) was comparable to that on placebo (1183 ng h/ml). Despite these drug-induced changes in plasma metoprolol concentration, neither cimetidine nor ranitidine altered the change in exercise-induced heart rate during dosing with metoprolol. PMID- 2979276 TI - Loss of predictive value of gastric ulcer symptoms in a randomized treatment trial. AB - Analysis of clinical data obtained in a double-blind randomized study, which compared liquid antacid (neutralizing capacity 120 mmol per day) with 1 g cimetidine in the treatment of 125 patients with gastric ulcer, revealed that, before starting treatment, 71% of the patients complained of epigastric pain, approximately 50% of bloating, and approximately 30% of nausea, heartburn, constipation or vomiting. Epigastric pain before treatment was significantly more frequent in patients with large ulcers (P less than 0.05) and in patients with ulcers unhealed after 4 weeks of therapy (P less than 0.05). This finding was the result of a highly significant correlation between diurnal epigastric pain and ulcer size and delayed healing (P less than 0.005). Nocturnal pain did not correlate with prognosis. In contrast to this correlation between pain before therapy and healing, the disappearance of epigastric pain with therapy did not signify ulcer healing. Only 14 (38%) of the 37 patients with healed ulcer were free from pain after the 4 weeks of therapy, whereas 25 (49%) of the 52 patients with persistent ulcers had no pain at this time. Placebo pain tablets relieved ulcer pain effectively in more than 85% of the patients, irrespective of whether the ulcer was healing or not. The other symptoms (bloating, nausea, heartburn, constipation or vomiting) were also alleviated by 4 weeks of therapy but no correlation was found with ulcer size or prognosis. The loss of the prognostic significance of ulcer pain is probably due to a complex interaction of the trial schedule on the patient's level of consciousness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979277 TI - The timing of evening meal and ranitidine administration--effects on patterns of 24 hour intragastric acidity. AB - Intragastric pH-metry was utilized to assess the effect of the time of meal ingestion and ranitidine administration on 24-h intragastric acidity. Twelve volunteers with a documented history of duodenal ulcer were studied in a four-way crossover design. Subjects randomly received ranitidine at 18.00 and 22.00 hours, with and without food. Serial blood samples were collected and analysed for ranitidine by high pressure liquid chromatography. Over the interval of 18.00 0.700 hours, the mean hydrogen-ion activity was significantly lower with the 18.00 hour dose than with the 22.00 hour dose (P less than or equal to 0.05). There were no differences between the four treatments in median pH or mean hydrogen-ion activity over the 23-h study interval. There were no differences between treatments in peak ranitidine concentrations, time to peak concentration, area under the serum-concentration time curve or elimination half-life. PMID- 2979278 TI - Should we use non-steroidal anti-inflammatory drugs? AB - Both the relative efficacy and inefficacy of non-steroidal antiinflammatory drugs (NSAIDs) contribute to their use in chronic rheumatic diseases. There are also sociological trends in patients, and in the population as a whole, increasing demand for treatment. In view of the risks of such treatment, the most rational approach to prescribing would be the use of a scientific risk-benefit analysis. Unfortunately, the data, especially those related to symptom relief, are inadequate for such an analysis. Until more meaningful figures are produced, good clinical practise concentrates on the responsibilities of physicians who both start and stop drugs, and makes it essential that strategies to minimize risk are produced. PMID- 2979279 TI - Diagnosis of gastrointestinal lesions during treatment with non-steroidal anti inflammatory drugs. AB - Probably all non-steroidal anti-inflammatory drugs (NSAIDs) increase the risk of gastrointestinal (GI) mucosal injury. The most frequent lesions are gastric erosions in the pre-pyloric region. Non-steroidal anti-inflammatory drugs also increase the incidence of peptic ulcers, although it is not yet clear whether more gastric ulcers or more duodenal ulcers are produced. Gastrointestinal symptoms are prevalent during NSAID treatment, but there is no correlation between symptoms and lesions. Clinical presentation may therefore vary considerably: many symptomatic patients will have no lesions, whereas others will present with complications but no symptoms. Blood loss is the most frequent complication of NSAID-induced GI lesions. Bleeding is usually 'silent' and occult. Overt haemorrhage, though rare, is more frequent in patients taking NSAIDs. Other complications, such as penetration and perforation, may also occur. Endoscopy is the diagnostic method of choice, as this method can detect even superficial mucosal lesions. However, because of the lack of correlation between symptoms and lesions, it is difficult to ascertain which patients are at risk and thus to avoid unnecessary diagnostic procedures. Future efforts should therefore be directed towards the prevention of NSAID-induced GI lesions. PMID- 2979280 TI - NSAIDs: a clinical approach to the problems of gastrointestinal side-effects. AB - Despite the fact that non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used drugs in medicine today, 2-10% of patients must discontinue their use primarily due to gastrointestinal (GI) side-effects. While the development of non-aspirin NSAIDs (NA-NSAIDs) has significantly reduced GI side effects, major problems persist. A practical clinical approach to these problems includes informing the patient about the risks and benefits of NSAIDs, risk management during treatment with NSAIDs and investigation of symptomatic side effects during treatment. Prophylaxis of GI side-effects is feasible in selected populations, but it has not been studied widely and may not be cost-effective. At present, costs of prophylaxis in all but selected populations with multiple risks probably outweight the benefits. PMID- 2979281 TI - Cost of treating arthritis and NSAID-related gastrointestinal side-effects. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are efficacious in the treatment of arthritis. However, side-effects particularly gastrointestinal toxicity, have been well documented with their use. Thus, in assessing total direct medical costs in the treatment of arthritis, the expenses involved in treating these side effects must be taken into account. A retrospective analysis was undertaken of all direct medical costs related to care of a group of Medicaid recipients treated for arthritis during a 2-year period. Data were obtained from the Medicaid Management Information System of Washington, D.C., USA. The actual expenditure of treating arthritis as well as the medical costs of associated NSAID-induced gastrointestinal side-effects were determined. Arthritis treatment costs per quarter were found to be $145 per patient. Approximately 25% of the population experienced NSAID-related gastrointestinal side-effects requiring further medical care. Treatment of these adverse effects costs an additional $66 per quarter per patient, thus adding 45.5% to the cost of arthritis treatment. Pharmaceutical claims comprised 42.4% of total adverse drug reaction treatment costs, while the few hospital claims accounted for 37.9% and physician charges 19.7%. It has been estimated that in 1983 direct medical costs of arthritis treatment in the USA was $8.6 billion. By extrapolating the costs in this study, it can be estimated that a further $3.9 billion was spent on treating gastrointestinal side-effects of NSAIDs, making a total of $12.5 billion. PMID- 2979282 TI - Ulcer and ulcer complications from non-steroidal anti-inflammatory drugs: what is the risk? AB - Gastroenterologists believe that non-steroidal anti-inflammatory drugs (NSAIDs) cause dyspepsia, may cause ulcers to develop de novo and cause ulcer bleeding and perforation. Regulatory authorities are aware that NSAID-associated adverse events are reported more often than for any other drug class, and that gastrointestinal events are most common and often serious. A case-control study in the UK indicates that those who use NSAIDs may be between two and four times as liable to gastrointestinal bleeding and probably perforation as non-users, particularly if elderly. It has further been suggested that the chances of dying of ulcer complications are very high in NSAID users. By contrast, studies in the USA conducted prospectively and post-marketing surveillance in the UK have appeared to show little risk. Differences may be partially, if not completely, explained by the variable methodologies employed. Dispute also exists about the rank order and significance of toxic effects among the various agents. PMID- 2979283 TI - Epidemiology of NSAID-induced gastrointestinal problems and the role of cimetidine in their prevention. AB - It is difficult to ascertain the incidence of gastrointestinal side-effects associated with intake of non-steroidal anti-inflammatory drugs (NSAIDs). In retrospective studies, some NSAIDs have been reported to be associated with a higher incidence of gastrointestinal side-effects than others. However, this has not been verified either in a prospective case-review study or in a large double blind study. Serious side-effects, such as bleeding, perforation and heart failure, occur in approximately 1% of patients using NSAIDs. One-third of all patients receiving NSAIDs will have gastrointestinal complaints. Since at least 10% of patients terminate treatment with NSAIDs as a result of side-effects, even reduction of those that are not life-threatening would be of great benefit. H2 receptor antagonists have proved effective in ulcer treatment, and their use as prophylaxis against the side-effects of NSAIDs is being widely studied. In a recent study, 63 patients who had experienced serious upper gastrointestinal side effects were given cimetidine while continuing their NSAID therapy. All but 4 of the 47 who had gastric or duodenal ulcer on first admission were healed at 8 weeks, and none of the remaining 16 with diffuse bleeding gastritis experienced further clinical episodes of bleeding or ulcer-related dyspepsia. PMID- 2979284 TI - Mechanisms of NSAID-induced gastrointestinal mucosal injury: a basis for preventing ulceration and symptoms from these agents. AB - Concepts are reviewed of the effects of anti-ulcer agents in counteracting the biochemical and cellular pathology of gastrointestinal injury by non-steroidal anti-inflammatory drugs (NSAIDs). While some measure of control of NSAID-induced mucosal damage is achieved in the upper gastrointestinal tract by the currently available anti-ulcer drugs, none can effectively counteract the major biochemical changes induced by NSAIDs. Some experimental approaches, which have been developed from recent studies in laboratory animals, are considered as a possible basis towards developing improvements in preventive procedures against NSAID induced gastrointestinal injury in man. PMID- 2979286 TI - Treatment of endoscopy-negative NSAID-induced upper gastrointestinal symptoms with cimetidine: an international multicentre collaborative study. AB - In this double-blind multicentre trial, 127 patients on non-steroidal anti inflammatory drugs (NSAIDs) who experienced gastrointestinal symptoms significant enough to warrant endoscopic evaluation, but without endoscopic evidence of mucosal damage, were randomized to receive cimetidine 400 mg b.d. or placebo for 4 weeks. Non-steroidal anti-inflammatory drug therapy was maintained at pre-trial doses for the duration of the study. After 4 weeks, 72% of cimetidine-treated patients experienced complete symptom resolution, as defined by a global symptom score of zero, compared with 49% on placebo. Complete disappearance of upper abdominal pain was achieved in 72% of the cimetidine-treated patients as opposed to 47% on placebo. Relief of heartburn was experienced by 87% of patients on cimetidine compared with 60% on placebo. Throughout treatment, the proportion of cimetidine-treated patients with nausea was consistently lower than in patients on placebo, while the two groups were similar with respect to disappearance of vomiting. At the completion of acute treatment, 27% of placebo-treated patients but only 12% of those on cimetidine were ineligible to continue maintenance due to persisting gastrointestinal symptoms. Patients who were symptom-free and required continuation of NSAID therapy were followed for 6 months during which they were re-randomized to receive cimetidine 400 mg nocte or placebo. Of the 105 patients included in this maintenance treatment phase of the study, 11% of the cimetidine-treated patients and 10% of those on placebo experienced relapse of symptoms. However, the mean time to relapse was 120 days with cimetidine compared with 35 days with placebo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979285 TI - Non-steroidal anti-inflammatory drugs and the gastric mucosa: mechanisms of damage and protection. AB - Prostaglandins have numerous mucosal protective properties, impairment of which contribute to NSAID injury. It is not clear which is the most important injurious factor affecting patients taking NSAIDs. Is it the NSAID itself, either by topical toxicity and/or production of toxic mediators or does the patient become more sensitive to NSAIDs as a result of some other luminal component, such as acid or bile acids, food (by physical or chemical mechanisms) or accompanying alcohol? Intrinsic irritancy/toxicity probably accounts for the greater mucosal toxicity of aspirin, although it is not clear whether this is due to a direct (non-prostaglandin-dependent) effect on mucus or bicarbonate secretion, membrane integrity, interference with intermediary metabolism or the production of toxic products. Similarly, the relationship of irritancy to the symptom of dyspepsia, to which patients on aspirin are prone, is as yet not well understood. While strategies for short-term protection of experimental animals and humans are well established, protection against important, but relatively rare clinical events, is more difficult to study and much less well understood. In particular, the importance of the ability of NSAIDs to exacerbate bleeding, independent of injury, has been largely neglected in the context of presentation with haematemesis and melaena. PMID- 2979287 TI - Treatment of NSAID-induced gastrointestinal lesions with cimetidine: an international multicentre collaborative study. AB - The efficacy of cimetidine 800 mg nocte in the treatment of erosions or ulcers induced by non-steroidal anti-inflammatory drugs (NSAIDs) was evaluated in an uncontrolled multicentre study of 187 patients requiring continuation of their NSAID therapy. After 4 weeks of treatment, endoscopic healing was achieved in 62% of patients. After 8 weeks of therapy, 88% of patients were lesion-free. Patients with ulcer had a healing rate of 49% at week 4, which increased to 81% at the completion of 8 weeks. The majority of patients (82%) with erosions healed in 4 weeks. By week 8, erosions were healed in 97% of patients. Following endoscopically verified lesion healing, 113 patients entered a maintenance phase of the study, which assessed the efficacy of cimetidine 400 mg nocte in preventing recurrence of erosions or ulcers while continuing NSAID therapy. During a 6-month observation period, the cumulative probability of endoscopically observed recurrence of lesions was 12% (with a mean time to recurrence of 116 days), which is similar to the incidence of relapse achieved with cimetidine 400 mg nocte in patients with uncomplicated peptic ulcer. The results of this study suggest that cimetidine 800 mg nocte is effective in healing NSAID-induced lesions despite continued NSAID use, and that maintenance treatment with cimetidine 400 mg nocte can prevent lesions and reduce lesion recurrence during chronic NSAID administration. PMID- 2979288 TI - Surgery in the management of the gastric side-effects of NSAIDs. AB - It is now accepted that the administration of non-steroidal anti-inflammatory drugs (NSAIDs) can result in peptic ulceration. Not infrequently, the first presentation of the ulcer is as a life-threatening complication such as a perforation or gastrointestinal bleed. Surgeons often become involved in management when one of these complications occurs. This paper discusses the ways in which NSAID-induced peptic ulceration may present to the surgeon and the special features that distinguish patients with NSAID-induced ulcers from the remainder of patients with peptic ulcer disease. It is pointed out how little has been written on the surgical management of this group of patients. The importance of the overall assessment of these patients, with the correction of nutritional deficiencies and the use of appropriate prophylactic therapeutic measures in their surgical management, is stressed. The criteria for determining what surgical procedure is used and results are described. PMID- 2979289 TI - Computed tomography and magnetic resonance imaging of the base of the tongue and the floor of the mouth. PMID- 2979290 TI - Correlative investigations of craniospinal anatomy and pathology with computed tomography, magnetic resonance imaging and cryomicrotomy. AB - A correlative computed tomographic-anatomic method was applied to multiplanar anatomic studies. The method was particularly valuable for comparative anatomic investigations of complex regions of the central nervous system. The description of CT and MR anatomy in this thesis is based either on direct CT-anatomic correlation of the same specimen, or on indirect MR-anatomic correlation with cryosectional images from cadavers. In sagittal partial saturation MR images with short repetition times, the pituitary fossa in 41 normal volunteers appeared inhomogeneous. A postero-inferiorly located high intensity signal correlated with an intrasellar fat pad in sagittal cryosectional images. The height of the pituitary gland in 38 normal volunteers was usually less than 8 mm and its upper surface was flat or concave. The cavernous sinus anatomy was studied in coronal and axial MR planes in seven normal volunteers and 15 patients in correlation with cryosectional images. The intracavernous cranial nerves were best shown in the coronal plane, in partial saturation and inversion recovery sequences and displayed as foci of high signals intensity. MR signs of a parasellar mass included obliteration of intracavernous venous spaces, displacement of the intracavernous portion of the internal carotid artery and bulging of the lateral wall of the cavernous sinus. The topographic anatomy of the cervical neuroforamina was investigated in axial, sagittal, coronal and oblique planes in a correlative CT-anatomic investigation in 19 specimens. In four normal volunteers, the surface coil MR images of the cervical neuroforamina were correlated with cryosectional images. Surface coil MR images in a plane perpendicular to the cervical nerve roots allowed to determine their relationship to intraforaminal structures and the boundaries of the foramen. The dorsal and ventral nerve roots were demonstrated with both CT and MRI. They were located in the lower half of the foramen at and below the intervertebral disc space. The dorsal nerve roots were in contact with the superior articular process. The ventral nerve roots abutted the uncinate process and the inferior portion of the foramen. In a biomechanical cervical spinal trauma study, experimental fractures were studied with CT in axial and sagittal planes. Sagittal anatomic images showed the fractures and soft tissue injuries. Non-displaced and horizontal fractures were generally difficult to detect on axial CT scans alone. In two pathologic spine specimens CT scans in axial, sagittal and coronal planes were compared with reformatted scans and cryosectional images. Direct CT images revealed four cervical spinal fractures whereas the reformatted images only showed one displaced pedicle fracture.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2979292 TI - Radioanatomy of the high jugular fossa and the labyrinthine portion of the facial canal. A radioanatomic and clinical investigation. PMID- 2979291 TI - Magnetic resonance imaging in malignant lymphoma. PMID- 2979293 TI - Inner ear structures and temporal bone pneumatization. A correlative radioanatomic investigation. PMID- 2979294 TI - Radiology in the diagnosis of adult respiratory distress syndrome with special reference to pulmonary oedema--its distribution and detection. PMID- 2979295 TI - Radiologic aspects of the diagnosis of supratentorial gliomas. An investigation by computed tomography, positron emission tomography and angiography. PMID- 2979296 TI - Evaluation of CT scanners for use in neuroradiology. A study with regard to radiation dose and image quality. PMID- 2979297 TI - Arachidonate cascade in the platelets of influenza virus infected mice. PMID- 2979298 TI - [A dynamic study on the values of Na, K and Cl in cadavers. A medico-legal case series]. PMID- 2979299 TI - Possible interaction between Hp and GLO systems. PMID- 2979300 TI - [Thanato-chemical considerations on death occurring at negative environmental temperatures]. PMID- 2979301 TI - Possibilities of examinations of group substances from skeletal remains of historical persons. PMID- 2979302 TI - [Thanatogenesis in acute pneumopathies (anatomo-clinical forms)]. PMID- 2979303 TI - Specification of colors in legal autopsy according to the CIE 1976 (L+ a+ b+) space. PMID- 2979304 TI - Phenotyping of esterase D in the Polish population by cellulose acetate gel isoelectrofocusing. PMID- 2979305 TI - Morphological changes in skin after laser irradiation. PMID- 2979306 TI - Ultrastructure changes in the liver after test exposure of laser irradiation. PMID- 2979307 TI - Difficulties in diagnosing diabetic coma in the process of a long-lasting diabetes in a state of intoxication. PMID- 2979308 TI - [Motivation in adolescent "gang" criminality. Analysis of group dynamics]. PMID- 2979309 TI - [Reconstruction of traffic accidents in legal medicine]. PMID- 2979310 TI - [Determination of paternity of a child born after artificial insemination]. PMID- 2979311 TI - [Legal medicine in state health system]. PMID- 2979312 TI - [Morphopathological considerations concerning inexplicable sudden death in children]. PMID- 2979313 TI - Morphological investigation of nodal arteries in sudden cardiac death. PMID- 2979314 TI - [Significance of red bone marrow in adults]. PMID- 2979315 TI - A function for inososes in mammalian tissue: inhibition of D-xylose: NADP 1 oxidoreductase and its analytical use for discrimination of inositols and inososes. PMID- 2979316 TI - Unusual circumstances of a road accident bearing on the opinion as to who was the driver. PMID- 2979317 TI - Solvent-abuse deaths--toxicological and morphological investigations. PMID- 2979318 TI - Topo-optical investigation of cardiac glycosides in the heart muscle. PMID- 2979319 TI - The usefulness of dactyloscopic foil for preserving traces of gunpowder substances in the vicinity of gunshot holes. PMID- 2979320 TI - From studies on karyolysis of cell nucleus. PMID- 2979321 TI - The first chair of forensic medicine of the University of Padua: Camillo Bonioli and Francesco Luigi Fanzago. PMID- 2979322 TI - Identification of blood stains from pregnant women by RIA determination of human chorionic gonadotropin. PMID- 2979323 TI - Comparative morphological investigation of cardiac glycoside intoxication. PMID- 2979324 TI - Cardiopathogenetic importance of detectable nickel in heart muscle after thermal injury. PMID- 2979325 TI - On diagnosis of a diabetic ketoacidosis post mortem (propane-2-on-/propane-2-ol state). PMID- 2979326 TI - Remarks on legal psychiatric aspects of the guardianship system for psychotics. PMID- 2979327 TI - Production and characterization of HLA antisera for paternity testing. PMID- 2979328 TI - Miniature ultrathin-layer isoelectric focusing in bloodstain analysis. PMID- 2979329 TI - Comparative morphological investigations to detect viruses: a clinico morphological study. PMID- 2979330 TI - [Scientific significance of dermatoglyphic examination in the framework of medico legal expert testimony in the study of filiation]. PMID- 2979331 TI - Death in police custody. PMID- 2979332 TI - Influence of legal regulations on the use of biological proofs in disputed paternity cases. PMID- 2979333 TI - [Dermatoglyphic studies in Hungary from the legal medicine viewpoint]. PMID- 2979334 TI - Potential sources of error in MNSs typing. PMID- 2979335 TI - Immunological identification of genotypes A and B blood groups. PMID- 2979336 TI - [Key enzymes of glycolysis in the rat brain. Regional differences and post mortem changes. Their application to the time of death]. PMID- 2979337 TI - [Death caused by Lyell's syndrome after prescription of pipemidic acid (Pipram): imputability and medico-legal problems]. PMID- 2979338 TI - Damage to lung parenchyma in laboratory animals drowned in various environments. PMID- 2979339 TI - [Medico-legal and social significance of the declaration of occupational disease status. Viewpoint based on concrete experience]. PMID- 2979340 TI - A follow-up study after twenty years on the relationship between air pollution and anthracosis. PMID- 2979341 TI - [Vitality diagnosis in lesions of the dermis in burns]. PMID- 2979342 TI - Unexpected difficulty in phenotyping red cell acid phosphatase (II). PMID- 2979343 TI - Sexual examinations done in the Medico-Legal Institute of Oporto (1980-1984). PMID- 2979344 TI - Sudden unexpected death as a complication of eating disorders. PMID- 2979345 TI - Intracerebral oxygen tension and carotid blood flow in asphyxial rabbits. PMID- 2979346 TI - [Ethics and medico-legal expertise]. PMID- 2979347 TI - Law and medical aspects connected with the use of pressure when taking blood for investigation purposes. PMID- 2979348 TI - [Medical ethics and expert physician: the expert encounter]. PMID- 2979349 TI - The ethics of medical expertise. PMID- 2979350 TI - Laser angioplasty-forensic aspects of a new therapeutic concept. PMID- 2979351 TI - Experiences of suicide attempts gained in the ambulance practice. PMID- 2979352 TI - An exceptional case of suicide through choking. PMID- 2979353 TI - Suicide in Portugal. PMID- 2979354 TI - A forensic pathologist in the surgery room: positive and negative issues. PMID- 2979355 TI - A suicide case of a drug addict. PMID- 2979356 TI - Suicides in Sap Vojvodina over the last 25 years. PMID- 2979357 TI - [Changes in suicidal deaths in Budapest during the 25 years from 1960 to 1984]. PMID- 2979358 TI - Suicide trends in Singapore. PMID- 2979360 TI - [Problems connected with social care provided to mentally retarded children and adolescents]. PMID- 2979359 TI - [Consent to experimentation: medical and legal duality]. PMID- 2979361 TI - The number 'one' case of 1984 in Greece. PMID- 2979362 TI - Psychiatric forensic aspects of the interaction between opiates and benzodiazepam. PMID- 2979363 TI - [The role of Regional Penitentiary Medico-Psychological Centers in the French system of aid to mental patients]. PMID- 2979364 TI - The reliability of information gained by a child's drawings. PMID- 2979365 TI - [Functional qualitative parameters of discernment]. PMID- 2979366 TI - [An interdisciplinary perspective of the medico-legal causality]. PMID- 2979367 TI - [Causal interpretation of the medico-legal reaction in the behavior]. PMID- 2979368 TI - [Methodological problems of legal-psychological expert testimony]. PMID- 2979369 TI - [Expectations of the court psychiatrist toward the psychologist in expert testimony concerning responsibility]. PMID- 2979370 TI - [Improper medical practices and expert testimony in medical responsibility]. PMID- 2979371 TI - Forensic psychiatric assessment of sexual offenders. PMID- 2979372 TI - Forensic psychiatric assessment of concussion and offences of drunken persons. PMID- 2979373 TI - Consequences for Swedish forensic psychiatry of a changed psychiatric legislation. PMID- 2979374 TI - Lack of motivation and personality alienation with respect to crimes committed in a drunken state. PMID- 2979375 TI - Some aspects of the forensic psychiatric examination in the case of young-age offenders. PMID- 2979377 TI - Essential facts of revealing the influence of manipulation of children. PMID- 2979378 TI - [Medico-legal psychiatry in Portugal]. PMID- 2979379 TI - [Micro-electronic technology, deviant behavior, bio-psycho-social considerations]. PMID- 2979376 TI - [Institutional observation at the open psychiatric ward in the criminal cases]. PMID- 2979380 TI - [Ethics and expert testimony: a specific example case, transsexualism]. PMID- 2979381 TI - [Prolegomena on discernment in deviant behavior related to consumption of alcoholic beverages]. PMID- 2979382 TI - [Discernment and dynamics of clinical configuration in current psychopathology]. PMID- 2979383 TI - [Criteria of the irresponsibility status in the psychiatric medico-legal expert testimony in Rumania]. PMID- 2979384 TI - Results and further possibilities of preventing suicides. PMID- 2979385 TI - [Ensemble of psychopathological symptoms, personality traits and circumstances in aggressive punishable offenses]. PMID- 2979386 TI - [Problem of a test method assessment of simulation and dissimulation in forensic expert testimony]. PMID- 2979387 TI - Legal and medical problems of drug addiction facing jurists and experts in forensic medicine. PMID- 2979388 TI - Lymphoma lymphociticum generalisatum and death under the clinical picture resembling PCP intoxication. PMID- 2979389 TI - Intraocular lens implantation from the medical insurance point of view. PMID- 2979390 TI - [Simulation, aggravation and dissimulation in the practice of ophthalmological medical expertise]. PMID- 2979391 TI - [Traumatology of fall from height]. PMID- 2979392 TI - Polytrauma in the praxis of medical experts. PMID- 2979393 TI - The importance of social policy in the protection of people's health in Poland. PMID- 2979394 TI - [Importance of legal medicine from the viewpoint of the attorney]. PMID- 2979395 TI - [Need to protect medical secrecy]. PMID- 2979396 TI - Fatal accidents in medical practice. PMID- 2979397 TI - Documentation and registration of the results of forensic examination as to the cause of death. PMID- 2979398 TI - Medico-legal aspects of anticoagulant therapy. PMID- 2979399 TI - A contribution to the problem of systemic fat embolism in the transplantation programme. PMID- 2979400 TI - Thinking about the problem of euthanasia. PMID- 2979401 TI - [Importance of social information in legal medicine]. PMID- 2979402 TI - [Legal aspects of the use of the computer in a hospital department]. PMID- 2979403 TI - Some ethical problems of medical practice due to the progress of science, technology and education. PMID- 2979404 TI - [Experimental medicine and ethics]. PMID- 2979405 TI - The rights of the patient and the possibility of refusing a treatment. PMID- 2979406 TI - Should the patient's rights of refusal override the doctor's Hippocratic Oath? PMID- 2979408 TI - [Legal medicine for the future]. PMID- 2979407 TI - [Autonomy of the sick at the junction of individual and social rights]. PMID- 2979409 TI - The integrity of the patient concerning medical treatment in Denmark. PMID- 2979410 TI - Public health, law, A.I.D.S. and human rights. PMID- 2979411 TI - Aggression and neglect of elderly persons. PMID- 2979412 TI - Human rights and medicine: for an ethical theory of integrity. PMID- 2979413 TI - I've always wanted to give a paper on ... ghosts. PMID- 2979415 TI - [Progress in determining the time of death]. PMID- 2979414 TI - Biochemical timing of wounds. PMID- 2979416 TI - [Alcoholism and the drivers' license]. PMID- 2979417 TI - Rules and regulations applied to organ transplantation in Belgium. PMID- 2979418 TI - [Removal of organs for therapeutic grafts. The state of legislation and its application in Italy]. PMID- 2979420 TI - Applications pratiques de la legislation sur les prelevements d'organes: les possibilites offertes sont-elles bien utilisees? [Practical applications of organ removal legislation. Are the possibilities well utilized?]. PMID- 2979419 TI - Organ removal from children and minors. Information and parents' consent. PMID- 2979421 TI - Ethical problems and legal regulations relating to the removal of tissue samples and organs from human corpses. PMID- 2979422 TI - The Spanish legislation on organ transplantation. PMID- 2979423 TI - [Legal and ethical rules on transplantation in Portugal]. PMID- 2979424 TI - [Legal, ethical and organizational aspects of organ transplantation in France]. PMID- 2979425 TI - [Organ transplantation: some legal and ethical problems on the international level]. PMID- 2979426 TI - Organ transplantation in Denmark. PMID- 2979427 TI - Determination of the time since death. III. Potassium in vitreous humour. Rise of precision by use of an "inner standard". PMID- 2979428 TI - [Body putrefaction in air-tight burials. I. Legislative aspect and synthesis of acquired scientific knowledge]. PMID- 2979429 TI - [Body putrefaction in air-tight burials. II. Attempt for a methodologic approach]. PMID- 2979430 TI - [Body putrefaction in air-tight burials. III. Macroscopic changes of cadavers and contamination flow]. PMID- 2979431 TI - [Body putrefaction in "air tight" burials. IV. Microbiologic findings and environment]. PMID- 2979432 TI - [Body putrefaction in "air-tight burials. V. Biochemistry of liquid and gaseous effusions]. PMID- 2979434 TI - Perspectives for the future of medico-legal entomology. Opening lecture of the group president. PMID- 2979433 TI - Post mortem correlation of urea and creatinine in different fluids. Special study of correlations between renin and angiotensin levels with agonal suffering and microscopical renal findings. PMID- 2979435 TI - Forensic entomology in Italy. PMID- 2979437 TI - Cadaveric fly activity in Maryland. PMID- 2979436 TI - Experimental reconstruction of Calliphora erythrocephala and Lucilia sericata life-cycles in the growth cabinet. PMID- 2979438 TI - Opiate analysis in cadaveric blowfly larvae as an indicator of narcotic intoxication. PMID- 2979439 TI - [Entomology applied to legal medicine. Its origins, its evolution]. PMID- 2979440 TI - [Insects and acari found in human cadavers in 49 medico-legal studies in Belgium]. PMID- 2979441 TI - [How to carry out an entomologic medico-legal assessment]. PMID- 2979442 TI - [Medico-legal entomologic assessment: several examples]. PMID- 2979443 TI - Medico-legal relevance of cadaver entomofauna for the determination of the time since death. PMID- 2979444 TI - [Influence of postmortem delay and duration of refrigeration on intra-ocular concentration of potassium]. PMID- 2979445 TI - [Assessment of mid-term postmortem delay by the determination of muscular creatinine]. PMID- 2979446 TI - [Evolution and current status of medico-legal entomology in Spain]. PMID- 2979447 TI - Medico-legal entomology at the meeting of Liege 1988 and its future. Synthesis and conclusions of the workshop 2. PMID- 2979448 TI - Studies on post mortem circulation of the blood. PMID- 2979450 TI - [Comparative analysis of postmortem glucose levels in blood and the vitreous humour]. PMID- 2979449 TI - Determination of glutathione and glutathione peroxidase activity after death. PMID- 2979451 TI - [Influence of postmortem time on the concentration of creatinine in the vitreous humour]. PMID- 2979453 TI - Determination of the time since death. I. Body heat loss and classical signs of death. An integrated approach. PMID- 2979452 TI - [Death dating by the study of nerve fibre demyelination of the median nerve]. PMID- 2979454 TI - Determination of the time since death. II. Electrical excitability of skeletal muscles. Current state and recent developments. PMID- 2979455 TI - [Patients' rights and the question of autonomy]. PMID- 2979456 TI - [Treatment refusal: a case study]. PMID- 2979457 TI - [From offense to sexual abuse in childhood. Legal aspects and ethical concepts]. PMID- 2979459 TI - Neuropsychiatry and medico-legal expertise. "New prospects in neuropsychiatry and the training of experts". Introductory remarks by Group President. PMID- 2979458 TI - Euthanasia and ethics. PMID- 2979460 TI - [Towards an objective methodology in psychopathology: contribution of new diagnostic systems of psychometrics and biological psychiatry]. PMID- 2979461 TI - [European community perspectives in the area of medico-legal psychiatric expert testimony]. PMID- 2979462 TI - [Introductory report to the Work Group. Patients' rights]. PMID- 2979463 TI - [Epilepsy and automobile driving]. PMID- 2979464 TI - [Mental capacity related to responsibility. A medico-legal interdisciplinary relational model]. PMID- 2979465 TI - [Mental capacity, penal capacity and capacity to use]. PMID- 2979466 TI - Training in forensic psychiatry in Sweden. PMID- 2979467 TI - [Problems of commitment of mentally ill in Spain]. PMID- 2979468 TI - Suicide in the psychiatric hospital. International trends and medico legal aspects. PMID- 2979469 TI - Impulsivity and attention-perception features in relation to juvenile delinquency. PMID- 2979470 TI - [Neuropsychiatric medico-legal expert testimony in Portugal]. PMID- 2979471 TI - On doctor's ethics. PMID- 2979472 TI - [Method of evaluating the discernment capacity according to the symptomatology]. PMID- 2979473 TI - [Reactive behavior with medico-legal implications. Interdisciplinary causal interpretation]. PMID- 2979474 TI - [Personality traits of delinquent and non-delinquent minors placed in an institution]. PMID- 2979475 TI - [Penal judiciary expertise. Harmonization of psychiatric and psychologic procedures]. PMID- 2979476 TI - Medico-legal and ethical approach to the legal interruption of pregnancy after aspiration biopsy of the chorionic villi and early amniocentesis. PMID- 2979478 TI - [Neuropsychiatric and medico-legal expert testimony as a specific mediator in the study of responsibility]. PMID- 2979477 TI - Drug delinquency and drug-related deaths in the Federal Republic of Germany and Western Europe. PMID- 2979479 TI - [Significance of the study of slow cerebral potentials in the psychiatric expert testimony in civil law cases]. PMID- 2979480 TI - [The treating physician, the psychiatrist and civil protection of incapable adults in France in 1988. What will be the evolution of the legislation after 1992?]. PMID- 2979481 TI - [AIDS and prison environment]. PMID- 2979482 TI - [Heroin addiction and HIV-1 virus. Report on consultations at the toxicologic clinic of the F. Widal Hospital]. PMID- 2979484 TI - [Bioethics and medical deontology: perspectives and contradictions]. PMID- 2979483 TI - [Diseases transmissible in prison environment with special reference to AIDS]. PMID- 2979485 TI - [Seropositivity in prison environment: deontologic and ethical aspects]. PMID- 2979486 TI - [Therapeutic abortion during the 3d trimester for fetal malformation. The position of the French law]. PMID- 2979488 TI - [Relations between the physician and the patient]. PMID- 2979487 TI - [What are the parents' rights to refuse care or prevention for their child? Medical and legal aspects]. PMID- 2979489 TI - [Medically assisted procreation and the Moslem law]. PMID- 2979490 TI - The right of the patient to refuse medical care. PMID- 2979491 TI - [Attitudes of physicians of the city of Murcia toward death and terminal-stage patients]. PMID- 2979492 TI - [Negligence toward the child and joint responsibility of the multidisciplinary team]. PMID- 2979493 TI - [Medico-legal aspect of the motto "my own master"]. PMID- 2979494 TI - [Injuries of the upper limbs. Evaluation of bodily injury]. PMID- 2979495 TI - [Psychometrics: methodology, reliability]. PMID- 2979496 TI - [Permanent absolute disability in the social security in Spain]. PMID- 2979497 TI - [Bodily injury. How is it possible to prepare harmonization of the evaluation of sequelae in the E.E.C]. PMID- 2979498 TI - [Evaluation of bodily injury of traumatized hand]. PMID- 2979499 TI - [Correlations, medical evaluation and compensation. Apropos of 57 cases of patients with severe skull injuries]. PMID- 2979500 TI - [Juvenile tort and compensation for bodily injury. The case of skull injuries]. PMID- 2979501 TI - [Bodily injury. Introductory report of the Chairman of the Work Group]. PMID- 2979502 TI - [For good use of medical certificate of the treating psychiatrist in expert testimony in common law or insurance]. PMID- 2979503 TI - [Is the independence of a medico-legal expert threatened?]. PMID- 2979504 TI - [Synthesis of the Work Group. Bodily injury]. PMID- 2979505 TI - [Compensation for personal injury in Italy: evaluation of the injury to health]. PMID- 2979506 TI - [Evaluation of the injury to the life of intercourse]. PMID- 2979507 TI - [Compensation for bodily injury in Italy]. PMID- 2979508 TI - [Bodily injury. Toward a unified European schedule]. PMID- 2979509 TI - [Search of uniform methods in the evaluation of bodily injury caused by traffic accidents]. PMID- 2979510 TI - [Medico-legal methodology in the evaluation of bodily injury in common law]. PMID- 2979511 TI - [Compensation of victims of therapeutic accidents]. PMID- 2979512 TI - [The physician-consultant of mutual insurers and bodily injury]. PMID- 2979513 TI - [Psychiatrization of expert testimony of bodily injury in Belgium]. PMID- 2979514 TI - Some comments on the compensation of bodily injury in the German Democratic Republic. PMID- 2979515 TI - Diel rhythms of tick parasitism on incubating African penguins. AB - 1. Ticks (Ornithodoros capensis Neumann) were most abundant on incubating host African penguins (Spheniscus demersus Linnaeus) at 24.00 hours and least abundant during 09.00-12.00 hours during 4 day periods in May and October at 3 h intervals. 2. Ticks were three times as abundant in May, the start of the breeding season, as in October, at its end. 3. Air temperature and humidity appear less important than light levels in determining tick activity. 4. The degree of tick parasitism of breeding seabirds is best studied at night. PMID- 2979516 TI - Differential effects of Bacillus sphaericus strain 2362 on Culex quinquefasciatus and its competitor Culex cinereus in West Africa. AB - The larval susceptibility to Bacillus sphaericus strain 2362 of the non-man biting mosquito Culex cinereus and the urban filariasis vector Culex quinquefasciatus, two competitor mosquitoes in polluted habitats, was compared. In the laboratory, both species ingested a similar amount of B. sphaericus spores when fed c. 2 x 10(5) spores per ml for 30 min. However, in the same experiment, third-instar larvae of Cx quinquefasciatus were reduced by 98% at 24 h exposure while Cx cinereus larvae were only reduced by 6% at 72 h. In the field, preimaginal populations of Cx cinereus ingested, within a week, more than 99% of the applied spores, but showed no significant decrease through 14 days in cesspools treated at 10 g/m2 of a flowable concentrate of B. sphaericus 2362, containing 2 x 10(10) spores/g. It is proposed that specific biological control of Cx quinquefasciatus could result from appropriate treatment of breeding-sites with larvicidal B. sphaericus and competitive displacement by Cx cinereus or other mosquitoes with larvae that are more tolerant of B. sphaericus. PMID- 2979517 TI - The Beffa form of Simulium soubrense of the S. damnosum complex in Togo and Benin. AB - The Beffa form of Simulium soubrense Vajime & Dunbar, a member of the S. sanctipauli sub-complex of the S. damnosum complex, was found breeding throughout rivers in the Togo-Benin Gap, as far north as 9 degrees 30'N. Its distribution changed with the season. In southern Togo there were seasonal fluctuations in the relative abundancies of the Beffa form and of S. damnosum/S.sirbanum. There was considerable temporal and regional variation in the frequencies of different colour morphs of adult flies. The flies in Benin tended to be darker. Infections with Onchocerca volvulus (Leuckart) appeared to be independent of the host's colour morph category. Larger flies harboured significantly more first stage Onchocerca larvae but no significant relations with fly size were found for second and third stage larvae. PMID- 2979518 TI - Absence of spirochaetes (Borrelia burgdorferi) and piroplasms (Babesia microti) in deer ticks (Ixodes dammini) parasitized by chalcid wasps (Hunterellus hookeri). AB - An entomophagous wasp (Hunterellus hookeri Howard) parasitizes about a third of the host-seeking nymphal Ixodes dammini Spielman et al. ticks on Naushon Island in Massachusetts (U.S.A.) where the agents of Lyme disease (Borrelia burgdorferi Johnson et al.) and human babesiosis (Babesia microti Franca) are enzootic. Following blood-feeding, wasp-parasitized ticks are destroyed by the developing wasp. The prevalence of either human pathogen in host-seeking ticks collected in wasp-infested sites is nearly 40% lower than that found in other sites. Nymphal ticks, collected early in their season of activity, are more frequently parasitized by the wasp and less frequently by the Lyme disease spirochaete than those collected later in the summer. Spirochaetes never infected wasp-infected ticks, and few wasp-infected ticks were concurrently infected by the Babesia piroplasm. Taken together, these correlations indicate that the wasp may render the tick inhospitable to both pathogens. The presence of the wasp may have reduced risk of human infection on the island by either pathogen by as much as a third. PMID- 2979519 TI - Experimental hut trials of permethrin-impregnated mosquito nets and eave curtains against malaria vectors in Tanzania. AB - Permethrin impregnated netting was tested against Tanzanian populations of Anopheles arabiensis Patton, An.gambiae Giles and An.funestus Giles in experimental huts fitted with traps to catch samples of the mosquitoes existing during the night. Treated bednets killed some mosquitoes and increased the tendency of survivors to exit during the night. Treated cotton did not perform so well as treated nylon bednets. An impregnated bednet in which holes had been cut, to simulate a torn net, reduced the number of mosquitoes which fed and survived approximately as well as an intact untreated net. Treated curtains around the eaves of experimental huts did not perform so well as bednets but caused considerable reductions in the number of mosquitoes which fed and survived. However, there was no such effect when treated netting was placed around the eaves of a dwelling house. When one child slept under a treated net and another slept outside the net in the same hut, the number of bites on the latter child was less than if neither child had been under a net. Various aspects of the applicability of permethrin impregnated nets on a community basis are discussed. PMID- 2979520 TI - Effect of host behaviour on host preference in Stomoxys calcitrans. AB - Field observations suggest that, in the U.K., cattle are the preferred host of Stomoxys calcitrans (L.), followed by horses. Differences were observed in the numbers of flies feeding on individual animals both in the field and under controlled conditions. Analysis of the behaviour of four Friesian calves under attack from S. calcitrans in controlled conditions revealed that the differences in the levels of attack between individual hosts are dependent on the reactions of the host when under attack. Those hosts which respond vigorously by tail flicks, foot stamps and head-swings suffer less from attack by S. calcitrans than their more placid contemporaries. PMID- 2979521 TI - Nectar feeding by mosquitoes in Sweden, with special reference to Culex pipiens and Cx torrentium. AB - 1. Nectar feeding by mosquitoes collected from tansy (Tanacetum vulgare) flowers was studied in July and August 1983-85 at two sites in central and south-western Sweden. Prior to fructose analysis, gonotrophic state and parity was determined. 2. A total of 1010 mosquitoes (70% males) of eighteen species was collected. Fructose was detected in 75% of the males and 78% of the females. Most (86%) mosquitoes collected belonged to Culex pipiens and Cx torrentium. 3. Among 219 Cx pipiens females, for most of which the abdomen appeared gravid (52%) or empty (42%), 81% were fructose-positive. Females of other species were also proved to have fed on plant sugars in both early and later stages of the gonotrophic cycle. 4. In Cx pipiens and males of Cx torrentium peak nectar feeding occurred between 22.00 and 04.00 hours. Data for males of both species suggested a V-shaped pattern of nectar feeding activity during the night. 5. At about 16 degrees C nearly all mosquitoes became fructose-negative in the anthrone test within 20 h after collection from tansy flowers. 6. At least 44% of Cx pipiens females that were attracted to a dove-baited trap had fed on plant sugars shortly before. PMID- 2979522 TI - Laboratory evaluation of cyromazine against insecticide-resistant field strains of Musca domestica. AB - A screening programme was undertaken to examine the possibility of cross resistance occurring between cyromazine and conventional insecticides. The responses of nine strains of Musca domestica to treatment with cyromazine, trichlorphon, methomyl and pyrethrins with piperonyl butoxide were measured. No tolerance to cyromazine was found, neither was there a direct correlation between larval responses to cyromazine and adult responses to other insecticides. Cyromazine is a potent larvicide against M. domestica and the results of these tests show that it has good potential for the control of houseflies with high levels of resistance to other insecticides. PMID- 2979523 TI - Climatological correlates of screwworm (Cochliomyia hominivorax) abundance in Texas, U.S.A. AB - The causes of screwworm (Cochliomyia hominivorax) outbreaks in North America are not well understood, but the literature suggests that climate was historically important. Screwworm case incidence in each of seven climatological divisions of Texas was examined for the years 1962-83, the period when sterile-male releases were made. Weak but statistically significant correlations were found between winter and summer cases and mean winter and summer rainfall and temperature when the independent variables were examined one at a time. Multiple regression of log case incidence on previous quarterly cases and current rainfall and temperatures showed a significant, negative effect of temperature on summer cases. Lagged screwworm cases accounted for most of the variation in quarterly cases. No climatic effects were detected in the other seasons. Rainfall was statistically unrelated to screwworm abundance in any season even in an arid region. The analysis does not support a climatic genesis of screwworm outbreaks or eradication. The sterile-male method is a credible explanation for screwworm disappearance. PMID- 2979524 TI - Two populations of Phlebotomus ariasi in the Cevennes focus of leishmaniasis in the south of France revealed by analysis of cuticular hydrocarbons. AB - Two distinct populations of Phlebotomus ariasi Tonnoir have been identified in the Cevennes focus of leishmaniasis in the south of France using gas-liquid chromatography (GLC) of cuticular hydrocarbons extracted from individual dried female flies. Results were obtained after analysis of flies collected from CDC light traps from a domestic and a sylvatic site separated by a distance of 900 m. Flies were provided for GLC analysis as six blind samples. Using cluster and discriminant analysis techniques, five of the samples were shown to form two distinct groups, while a sixth was identified as a mixture. These findings were subsequently confirmed to correspond exactly with the way the samples had been presented. Samples grouped together on the basis of the flies' cuticular hydrocarbon profiles had been taken from the same site, while the ungrouped sample had been deliberately mixed. Using a jack-knifed estimator, it is shown that specimens can be correctly allocated to the population to which they belong with a 92% success rate. These results confirm the value of the technique for the identification of populations of medical vectors. The implication of the findings for the epidemiology of leishmaniasis is discussed with special reference to the need to determine if both populations are vectors and to study differences in behaviour. In addition to adults, profiles of larval sandflies have been obtained for the first time. PMID- 2979525 TI - The larvae of some blowflies of medical and veterinary importance. AB - Diagnostic features are described as a series of couplets that enable separation of the third instar larvae of the following pairs of closely related forms of blowflies of medical and veterinary importance: Chrysomya chloropyga (Wiedemann) and Ch.putoria (Wiedemann), Chrysomya albiceps (Wiedemann) and Ch.rufifacies (Macquart), Cochliomyia hominivorax (Coquerel) and Co.macellaria (Fabricius), Lucilia sericata (Mergen) and L. cuprina (Wiedemann), Calliphora augur (Fabricius) and C. stygia (Fabricius). PMID- 2979526 TI - DNA probes for species identification of mosquitoes in the Anopheles gambiae complex. AB - Identification of species within the Anopheles gambiae Giles species complex is essential for the correct evaluation of malaria vector ecology studies and control programmes. The development of DNA probes to distinguish species of the An.gambiae complex is described. Genomic libraries were prepared for four members of the An.gambiae complex. These were screened using radiolabelled DNA from different species of An.gambiae sensu lato and a number of clones selected on the basis of their species specificity. These clones could be divided into two groups, each containing homologous sequences. Sequences homologous to group 1 inserts are highly reiterated in the genomes of Anopheles arabiensis Patton and Anopheles merus Donitz, present in low copy number in Anopheles melas Theobald, but were not detected in Anopheles gambiae sensu stricto. Studies on the organization of this sequence in the genome of An.arabiensis show that homologous sequences are male specific and interspersed within the chromatin. Sequences homologous to group 2 inserts are highly repeated in the genomes of An.merus and An.melas, but present in low copy number in An.gambiae s.s. and An.arabiensis. Group 2 homologous sequences are not sex-specific in the species tested and appear to be tandemly repeated. When used as hybridization probes, these sequences provide a sensitive means for the identification of species within the Anopheles gambiae complex. PMID- 2979527 TI - Management of insecticide resistance in control of the Simulium damnosum complex by the Onchocerciasis Control Programme, West Africa: potential use of negative correlation between organophosphate resistance and pyrethroid susceptibility. AB - 1. Resistance of some populations of the Simulium damnosum complex to temephos (100-fold at the LC50 level), with degrees of cross-resistance to chlorphoxim (14 fold) and other organophosphate insecticides, follows intensive larvicidal control of S. damnosum s.l. in West African river systems since 1975 by the WHO Onchocerciasis Control Programme. 2. Larvae of at least three sibling species of the S. damnosum complex have become organophosphate-resistant: these are the forest species S. sanctipauli Vajime & Dunbar and the savanna species S. sirbanum V. & D. and S. damnosum Theobald sensu stricto. 3. Organophosphate-resistant S. damnosum s.l. larvae show increased susceptibility to some organochlorine and pyrethroid insecticides, especially to permethrin (up to 11-fold) and OMS 3002 (up to 17-fold), as compared with organophosphate-susceptible populations. 4. This differential susceptibility is reflected by increased pyrethroid efficacy in operational use for river treatments against organophosphate-resistant field populations of S. damnosum s.l. larvae. Treatment of 100 km of the lower Bandama River in 1985 showed that permethrin at the highly selective dosage of 10 min exposure to 0.01 mg/l caused reversion towards organophosphate-susceptibility of the target population of S. sanctipauli. This effect was less pronounced when the Comoe River was treated at the lower dosage of 0.005 mg/l for 10 min. 5. To overcome temephos-resistance, it is proposed that the most rational usage of currently available larvicides would involve the following annual sequence of treatments: Bacillus thuringiensis serotype H-14 when river discharge is below 75 m3/s; chlorphoxim for about eight weekly treatment cycles after river discharge rises; permethrin (or alternative pyrethroid) for up to six treatment cycles- this should eliminate any incipient selection for chlorphoxim-resistance; resume chlorphoxim (or perhaps carbosulfan) treatments until river discharge falls below 75 m3/s permitting resumed use of B.t. H-14. PMID- 2979529 TI - Efficacy of Bacillus sphaericus 2362 against larvae of Anopheles gambiae under laboratory and field conditions in West Africa. AB - A flowable concentrate of Bacillus sphaericus (Neide) strain 2362 was applied against Anopheles gambiae Giles s.l. mosquito larve in small plot field trials in Bobo-Dioulasso area. Burkina-Faso. Third and fourth instar larvae were controlled for 10-15 days with a dosage of 10 g/m2, 3-10 days with 1 or 0.1 mg/m2, and 2 days with 0.01 g/m2. Complete elimination of larval populations required 1 x 10(2) to 2 x 10(3) viable spores/ml in the larval feeding zone. After treatment, the total numbers of viable spores decreased in the ponds, due to ingestion of spores by non-target as well as target organisms and/or loss of viability of some spores by sunlight. This formulation was less effective against An. gambiae than against Culex quinquefasciatus Say larvae, both in laboratory bioassays and under field conditions. PMID- 2979528 TI - Base-line susceptibility of Phlebotomus papatasi to insecticides. AB - Susceptibility of the leishmaniasis vector sandfly Phlebotomus papatasi (Scopoli) to three insecticides was assessed on a colony originating in the Jordan Valley, Israel. Results are considered to show susceptibility to DDT and permethrin. The results for methoxychlor indicated the possibility of heterogeneity in the population and tolerance towards this compound. PMID- 2979530 TI - Mites from house dust in Glasgow. AB - Glasgow's mild, high-rainfall climate, combined with a deteriorating quality of housing and low standards of living in many parts of the city, makes it a particularly suitable place for thriving populations of house dust mites. The acarofauna in 124 samples of house dust from beds and carpets in seventy-four homes in Glasgow, Scotland, comprised thirty-one species of which the most abundant were Dermatophagoides pteronyssinus (Trouessart) (64.3%), Glycyphagus domesticus (De Geer) (16.7%), Euroglyphus maynei (Cooreman) (11.6%), Tarsonemus sp. (1.6%), Cheyletus eruditus (Schrank) (1.5%), C. trouessarti Oudemans (0.9%), Tarsonemus fusarii Cooreman (0.8%) and Glycyphagus destructor (Schrank) (0.7%). Mites were present in all the homes surveyed and the mean population density was found to be 97/100 mg of dust (range 2-1210). Over 47% of homes visited showed signs of disrepair associated with damp, especially unmodernized flats in old tenement buildings and 1960s council housing stock, many of which contain deprived occupants. There was a high incidence of hygrophilic species such as Glycyphagus spp., Tarsonemus spp. and Euroglyphus maynei in such homes. Samples from homes of atopic asthmatics were found to contain significantly fewer mites than those from normal volunteers (chi 2 = 54.7). This was partly due to the use of house dust mite avoidance measures (e.g. regular vacuum cleaning of mattresses as well as carpets) by some of the asthmatics. PMID- 2979531 TI - Efficacy of permethrin-impregnated curtains for malaria vector control. AB - Preliminary results obtained by the use of permethrin-impregnated curtains against the Afrotropical malaria vectors Anopheles gambiae Giles s.l. and An.funestus Giles are reported and discussed. Field trials were carried out in villages near Ouagadougou, Burkina Faso. Houses were provided with curtains made from 100% cotton netting, impregnated with permethrin at the dose of 1 g a.i./m2, to cover the doorway, the window(s) and the space under the eaves. Entomological data collected during the period 1985-86 showed residual permethrin activity for about a year, and almost complete prevention of indoor-resting mosquitoes. Increased exit-rate and mortality-rate of house-entering malaria vectors were also obtained. Utilization of this malaria vector control method in primary health care programmes is advocated. PMID- 2979532 TI - Experimental hybridization between members of the Simulium damnosum complex. AB - Males of the Beffa form of Simulium soubrense Vajime and Dunbar were successfully crossed with female S.squamosum V. & D. and S.sanctipauli V. & D. Fertile eggs from both crosses were reared to larvae and, in the case of the Beffa x squamosum cross, to adults. Larval chromosomes from the Beffa x sanctipauli cross had normally paired homologues but those of the Beffa x squamosum cross showed extensive asynapsis and large heterozygous loops. The morphology of the larval and adult hybrids from the Beffa x squamosum cross were more characteristic of squamosum, but the adult males had scutal patterns typical of the Beffa form and retained this form's laboratory-mating propensity. PMID- 2979533 TI - Development of a system for sterilizing tsetse flies, Glossina spp., in the field. AB - Various autosterilizing systems were evaluated on natural populations of Glossina pallidipes Austen and G.morsitans morsitans Westwood in Zimbabwe. These involved a clear plastic tower of two or three chambers mounted at the cage position on a trap. Inside one chamber flies could be exposed to the vapour phase of the chemosterilant bisazir, P,P-bis (1-aziridinyl)-N-methylphosphinothioic amide. The system was designed to encourage flies to enter the sterilizing chamber, delaying their exit for sufficient time to expose them to a sterilizing dose. This was accomplished in the most effective system by restriction of the exit to one small hole (6 mm diameter) at the roof-side junction. The number of flies remaining in the chamber declined exponentially. The rate of exit was directly proportional to the density of flies in the sterilizing chamber and to the number of exit holes. The probability of a fly being in this chamber for at least 1 or 7 min (the times taken for female and male G.m.morsitans respectively to receive an ED50) was 0.84 and 0.67 respectively with one fly present. With sixteen flies, these probabilities were 0.76 and 0.18 respectively. Results suggest that it may be possible to develop a cheap, safe and efficient autosterilizer for use on tsetse traps. PMID- 2979534 TI - A new type of collapsible insect-surveillance light trap for sampling Diptera. AB - The newly designed Army Collapsible Insect Surveillance trap for nocturnal collection of flying Diptera that are attracted by light is described. A polyvinyl chloride construction increases durability, while the capability to collapse from a height of 92.1 cm to 24.1 cm facilitates transportation and storage. PMID- 2979535 TI - The status of Simulium oyapockense and S. limbatum as vectors of human onchocerciasis in Brazilian Amazonia. AB - In an attempt to explain the current distribution of onchocerciasis in the forests of northern Brazil (Moraes et al., 1979, 1986), and its potential for dispersal to other areas, this study compares the vector status of Simulium oyapockense Floch and Abonnenc, 1946 in both a hypoendemic and an onchocerciasis free area with that of S. limbatum Knab, 1915 in the latter area. Both species allowed the full development of Onchocerca volvulus (Leuckart) to the infective L3 stage after experimental infection with microfilariae. Their vector competence was significantly lower than for other efficient vector species in South America and Africa because of the lethal effect of the cibarial armature on ingested microfilariae. The low vector capacity of S. oyapockense, together with the low prevalence and intensity of infection of O. volvulus, probably explains why onchocerciasis has not significantly increased in intensity over the last 10 years in the hypoendemic part of the Amazonian focus. Omnipresence of both vector species in the adjacent savanna region, however, could facilitate the spread of onchocerciasis if human population movements continue from the hyperendemic part of the onchocerciasis focus. PMID- 2979537 TI - Septic tank mosquitoes: competition between species in central Nigeria. AB - Exit traps, placed over the air vents of septic tanks, were used to examine species diversity and relative abundance of mosquitoes breeding in ammonia-rich waters of septic tanks. Of the six species found, Culex decens Theobald and Culex cinereus Theobald appeared to be competing successfully with Culex quinquefasciatus Say during the wet season but not during the long dry season. The seasonal timing of their displacement by Cx quinquefasciatus was variable and did not correlate well with climatic factors. The three other species present, generally during the wet season and early dry season, were Culex tigripes G. & C., Culex horridus Edwards and Aedes aegypt (L.). Experimental bucket ovitraps were used to assess preference towards covered (dark) septic tank water in comparison with sunlit septic tank water, covered and sunlit compost water. These were colonized by Cx quinquefasciatus, Cx decens, Ae. aegypti and Ae. vittatus Bigot. The covered septic tank water was more abundantly colonized by Cx quinquefasciatus and marginally so by the two Aedes species. Cx decens appeared to colonize the exposed compost water more readily in the dry season, but changed to the covered septic tank water in the wet season. The discussion centres around competition between these mosquitos species and concludes that it would be useful to know what environmental factors, or what aspects of competition, lead to severe natural reductions in the abundance of the major pest species Cx quinquefasciatus. PMID- 2979536 TI - Trypsin and chymotrypsin-like enzymes of the sandfly Phlebotomus papatasi infected with Leishmania and their possible role in vector competence. AB - Phlebotomus papatasi (Scopoli) is susceptible to infection with Leishmania major Yakimov & Schokov and resistant to L. donovani Laveran & Mesnil. The possibility that susceptibility depends on midgut levels of trypsin and chymotrypsin-like (esterolytic) enzymes was investigated. Infection with L. major reduced the trypsin-like activity to 93.5% and 86% of the control value at 20 and 30 h post feeding and increased it to 106% at 52 h. Infection with L. donovani reduced trypsin-like activity to 64% and 73% of the control value at 30 and 52 h post feeding. The overall amount of trypsin and chymotrypsin-like enzymes in L. major infections was reduced to 50% and 34% of the control value at 20 and 30 h post feeding and increased to 184% at 52 h. Only one of the enzymes separated by gel electrophoresis was lower throughout, i.e. peak D. Overall, the midgut enzyme level with L. donovani infection was 86% of the control value at 30 h post feeding and 105% at 52 h; their relative amounts changed throughout. Soybean trypsin inhibitor enabled L. donovani to survive and multiply in P. papatasi. It is suggested that a specific component of the trypsin-like activity prevents the survival of L. donovani in P. papatasi and that modulation of this factor enables L. major to survive. PMID- 2979538 TI - Proteolytic activity in the ectoperitrophic fluid of blood-fed Culex nigripalpus. AB - In blood-fed Culex nigripalpus Theobald, proteolytic activity appeared in the ectoperitrophic fluid after 3 h, but only after 6 h in a homogenate of the blood filled midgut. The activity continued to be higher in ectoperitrophic fluid than in whole gut homogenate until about 40 h after the meal, when most of the intact clot had disappeared. Apparently, undigested blood inhibits proteolytic activity. The blood clot lacked activity and the inhibitor could not be removed by washing. The results are compatible with a hypothesis that the peritrophic membrane separates the digestion from the ingestion compartment. PMID- 2979539 TI - Tsetse ecology in a Liberian rain-forest focus of Gambian sleeping sickness. AB - Investigations on tsetse ecology were undertaken in Bong County of Liberia during the dry season, October 1981 to February 1982, around villages where the human infection rate with Trypanosoma brucei gambiense Dutton was about 2%. Most tsetse captured in biconical traps were Glossina palpalis Robineau-Desvoidy and G. pallicera Bigot, with relatively few G. fusca Walker and G. nigrofusca Newstead. Swamps and water-gathering places were predominant habitats of all four species, but tsetse were also found in coffee and cocoa plantations. Breeding-places of G. palpalis were found in the leaf axils of oilpalm trees (Elaeis guineensis Jacquin), especially beside paths where people would risk being bitten. Bloodmeals of twenty-nine wild-caught G. palpalis were identified as mostly from man (fifteen) and bushbuck (Tragelaphus scriptus (Pallas] or other wild ruminants (eleven), plus three from reptiles. It is concluded that man may be the principal host of tsetse in the area, while man or bushbuck could be the main reservoir to T.b. gambiense infection. Most of the activity of G. palpalis occurs in the early afternoon from noon to 16.00 hours. Mean life-span of G. palpalis and G. pallicera, estimated from wing-fray age-groups, was consistent with the females, and to a lesser degree the males, having vector potential. PMID- 2979540 TI - Egg-float ridge number in Anopheles stephensi: ecological variation and genetic analysis. AB - Eight Indian laboratory stocks of Anopheles stephensi Liston could be grouped into three categories with, respectively, 14-22, 12-17 and 9-15 ridges on the egg floats. The mode number of ridges among the eggs laid by individual females in these stocks was 16-19, 13-16 and 10-14, respectively. The category with the highest egg-float ridge number corresponded with the type-form and the lowest with var. mysorensis Sweet and Rao; the new egg-float category with ridge number modes of thirteen to sixteen was designated as 'intermediate'. All three forms, i.e. type-form, intermediate and myosorensis were observed in semi-urban areas while only intermediate and mysorensis were seen in rural areas. Breeding experiments indicated no post-copulatory barriers between the populations. Likelihood analysis of the results of crosses and back crosses indicated that variation in ridge number is controlled by more than one genetic factor. The stocks with different ridge numbers are best considered as 'ecological variants'. PMID- 2979541 TI - Evaluation of some adhesives for collecting Musca domestica and Chrysomya megacephala adults or mosquito larvae in sticky traps. AB - 1. Seven types of water-insoluble adhesives were evaluated in sticky traps for collecting adults of Musca domestica L. and Chrysomya megacephala (Fabricius) or mosquito larvae (Aedes aegypti (L.) and Culex quinquefasciatus Say). 2. Adhesive viscosity affected the tackiness of the glues and this determined their trapping efficiency in air or water. 3. From the 'Hyvis' range of adhesives tested, 'Hyvis 200' was most effective for trapping adult flies. 4. With 24 h exposure to fourth instar Ae.aegypti larvae in tapwater, submerged plates coated with 'Hyvis 10', 'Hyvis 30' or 'Hyvis 200' formulations trapped the majority of larvae. In polluted water the highest rates of trapping were 17.3% of Ae.aegypti and 18.7% of Cx quinquefasciatus with 'Hyvis 200'. Floating traps were consistently less productive than submerged traps under laboratory conditions. 5. In a heavily polluted natural breeding-site of Cx quinquefasciatus, floating traps were more productive than submerged sticky traps with four of seven adhesives tested, the most efficient being 'Hyvis 200' (4.2 mosquitoes per hour) and Hyvis:polyethylene 90:10 (4.5/h). Despite the relative inefficiency of aquatic traps, emergent adults, pupae and second to fourth instars of larvae were collected quickly from the habitat. PMID- 2979542 TI - Morphological aspects of the third instar larva of Haematobia irritans. AB - The main morphological features of the cephalic region of the larva of Haematobia irritans (L.) are the oral grooves, tripartite labium and the antennomaxillary protuberances that have the dorsal, terminal and ventral sensory organs. The total number of sensilla that are found on the terminal organ differs from other cyclorrhaphous-fly larvae. The fan-shaped anterior spiracles usually consist of seven bulbous digits that are unequal in length. The creeping welts consist of notched, convex plates that split into two separate plates as they approach the midline of the venter. This characteristic has not been described previously for this species or other, higher, dipterous larvae. There are two posterior spiracles with an ecdysial scar, four fan-shaped and branching spiracular hairs and irregularly-shaped spiracular openings. The longitudinal anal opening is situated in the cuticular band that is known as the anal organ. PMID- 2979543 TI - Suppressive action of Samorin on the cyclical development of pathogenic trypanosomes in Glossina morsitans centralis. AB - Male Glossina sexually sterilized by gamma-irradiation are as efficient vectors of trypanosomiasis as fertile males. An attempt was made, using isometamidium chloride (Samorin), to interfere with the cyclical development of trypanosomes in sterile males, destined for use in the sterile insect release (SIR) method of tsetse eradication. The infection rate with mature Trypanosoma congolense Broden was effectively reduced in sterile male Glossina morsitans centralis Machado, when the flies were fed on an infected goat 2 days after they were fed as tenerals on an in vitro bloodmeal containing 8 micrograms Samorin/ml blood. The infection rates with mature T.vivax Ziemann and T.brucei brucei Plimmer & Bradford were completely suppressed at this drug dose. Whensterile teneral males were fed on a bloodmeal containing 12 micrograms/ml Samorin and given the infected bloodmeal 10 days later, infections by mature T.vivax, T.congolense and T.b.brucei were completely suppressed. Hence in the management of a tsetse eradication programme utilizing the SIR method, it is recommended that the sterile teneral male tsetse should, prior to release, be given a bloodmeal containing Samorin at 12-15 micrograms/ml blood. This will effectively suppress future disease transmission. PMID- 2979544 TI - Field trial of the effectiveness of indoor-spraying with pirimiphos-methyl emulsion for malaria control in a tribal area of Phulbani district, Orissa State, India. AB - A field trial of malaria vector control was conducted in Phulbani district, Orissa, during 1984 and 1985. Indoor-spraying of pirimiphos-methyl emulsion formulation was undertaken at an application rate of 2 g/m2 in two sections (population 14,692) of Nuagaon Primary Health Centre. Houses in two adjacent sections (population 21,450) were sprayed with DDT a water dispersible powder (wdp) formulation at 1 g/m2 for comparison purposes. Operational problems in this area come from the tendency of tribal people to re-plaster over wdp applications. Pre-spray malariological indices in the trial area were 38% slide positivity rate, 37% slide falciparum rate and 12.1% annual parasite incidence. Densities of Anopheles annularis Van der Wulp, An. culicifacies Giles, An. fluviatilis Theobald and other potential malaria vectors were reduced in the pirimiphos methyl trial area 2-35-fold more than in the area sprayed with DDT. Malariological indices were reduced by 65-68% in the pirimiphos-methyl sprayed area compared with only 26-35% reduction in the DDT sprayed area. Spraymen and villagers experienced no adverse side-effects from residual house-spraying with pirimiphos-methyl emulsion and it is concluded that this organophosphate product has advantages for malaria vector control, especially in operationally difficult situations. PMID- 2979545 TI - Honeydew of aphids as a source of sugar for Phlebotomus ariasi. AB - Wild-caught Phlebotomus ariasi Tonnoir, starved until their sugar meals had been digested, were caged for 24 h with plants or plants infested with aphids and then tested for fructose, a constituent of plant sap. No evidence was found that the flies took sap directly from nine types of plants present in their habitat. About two-thirds of flies caged with oak (Quercus ilex L.) infested with an aphid [Lachnus roboris (L.)] and about a fifth kept with leaves of the french bean (Phaseolus vulgaris L.) infested with an aphid took honeydew. Experiments with three other aphid species gave negative results. Of sandflies caught in an oak tree infested with aphids, half of the females and three-quarters of the males contained fructose. Tests on females caught in a house and grouped according to state of engorgement showed that, having taken blood, the females stop taking sugar until the bloodmeal is completely digested. It is predicted that honeydews are important in the development of Leishmania infantum Nicolle in the gut of P. ariasi. PMID- 2979546 TI - Observations on the Anopheles gambiae complex in the Senegal River Basin, West Africa. AB - 1. Three sibling species of mosquitoes of the Anopheles gambiae complex are found in the Senegal River Basin: An. melas Theobald, An. gambiae Giles and An. arabiensis Patton. 2. An. melas is restricted to the river delta and environs where saltwater breeding places are present. 3. An. gambiae and An. arabiensis are sympatric in the study area; An. arabiensis predominates in coastal zones where it breeds also during the dry season; An. gambiae predominates in inland areas where breeding is mostly restricted to the rainy season (July-September). 4. An. arabiensis is chromosomally polymorphic all over the study area, with much variation of inversion frequencies, particularly for the 2Ra arrangement. 5. An. gambiae is characterized by a very high frequency of the 2Rb-2La inversion arrangement which is typical of the Savanna chromosomal form. PMID- 2979547 TI - A behavioural bioassay to identify attractive odours for Glossinidae. AB - 1. A behavioural bioassay, based on antennal movement responses, was developed using Glossina morsitans morsitans Westwood for screening chemical attractancy to tsetse. 2. Chemicals found to be attractive to male tsetse were acetone, formaldehyde, methylethylketone, methylvinylketone, 1-octen-3-ol and pentanal but not acetophenone, hexanal, lactic acid or urea. 3. Female tsetse also responded to all these chemicals in a similar fashion. Overall responses of females were, however, less than those of males. 4. These laboratory bioassay findings agree with field observations on tsetse responses to certain chemical odours. Therefore this behavioural bioassay should be a useful laboratory test procedure for screening attractants. PMID- 2979548 TI - The effect of permethrin-impregnated bednets on a population of Anopheles farauti in coastal Papua New Guinea. AB - The effect of introducing bednets impregnated with 0.4 g/m2 permethrin on local populations of the malaria vector mosquitoes Anopheles farauti Laveran and An. koliensis Owen was monitored in a coastal village of Papua New Guinea. Whole night landing collections were undertaken for 25 consecutive nights before and 21 nights after the introduction of the nets. Capture-recapture experiments and resting collections were also performed before and after the introduction of the nets. Following the introduction of treated nets, the biting population of An. farauti, the predominant vector, dropped from an average of 689 to 483 per man night and the oviposition cycle became irregular, although survival rates (determined by time series analysis of the landing catches and log regression of recapture rates) were not significantly affected. The densities of An. farauti resting in and around houses and the human blood index of the engorged females also decreased significantly after introduction of the treated bednets. The population of An. koliensis dropped prior to the introduction of the nets. However, the number of nulliparous females in the landing catches remained more or less constant which implies that, in this species, survival rates were affected by the nets but that recruitment to the population was not. PMID- 2979549 TI - The effect of Brugia pahangi infection on survival of susceptible and refractory species of the Aedes scutellaris complex. AB - Life table statistics were used to examine the survival functions of filarial susceptible and refractory species of the Aedes scutellaris (Walker) group of mosquitoes, following infection with high and moderate doses of Brugia pahangi (Buckley & Edeson). Survivorship curves and hazard function curves were generated, and the median survival times and the proportions of mosquitoes surviving beyond the extrinsic incubation period of the parasite were determined. In the susceptible populations of Aedes polynesiensis Marks, Ae. pseudoscutellaris (Theobald) and Ae.tabu Ramalingam & Belkin a dose-response relationship was detected between parasite load and mortality. This relationship was characterized by a significant reduction in the proportions of infected female mosquitoes surviving at days 1 and 9 postinfection, reduction in the median survival times and an increase in the hazard rates as the infectious dose increased. The survival of the refractory species, Ae.alcasidi Huang and Ae.katherinensis Woodhill was not significantly affected by the infection. A positive correlation between microfilaraemia in the vertebrate host and parasite load in the susceptible mosquito populations was also observed. Regression analysis of the number of parasites recovered from susceptible mosquitoes at the time of death showed that mosquitoes at highest risk of dying harboured from 11.6 to 19.4 infective larvae when fed on a gerbil with sixty-five microfilariae per 20 microliters blood; this resulted in 34.4-40.2% mortality by day 9 postinfection. A mean number of 32.6-46.9 infective larvae was observed when these populations were exposed to a gerbil with a microfilaraemia of 150 mf/20 microliters and resulted in 72.8% to 80% mortality in these populations. Viable infective larvae were recovered from infected mosquitoes up to 50 days postinfection. PMID- 2979550 TI - Leishmania infantum infection rates in Phlebotomus perniciosus fed on naturally infected dogs under antimonial treatment. AB - Dogs naturally infected with Leishmania infantum Nicolle were treated with three courses of meglumine antimoniate. Changes were observed in the dogs' clinical signs, antibody titres and in infection rates of Phlebotomus perniciosus Newstead fed on the dogs. A large reduction in the sandfly infection rate was observed for 4-5 months after the first treatment. The use of antimonial drugs is advocated for the control of canine leishmaniasis and to reduce risks of L.infantum transmission. PMID- 2979551 TI - Geographical distribution of Glossina palpalis gambiensis and G.p.palpalis in Liberia. AB - The two subspecies of Glossina palpalis (Robineau-Desvoidy) occurring in Liberia could be reliably separated morphometrically by measuring the width of the terminal dilatations of the male inferior claspers. Subspecies differentiation of female flies was less conclusive. Identification of flies from fifty-four sites revealed that most of Liberia lies in the belt of Glossina palpalis palpalis. However, pure and substantial populations of G.p.gambiensis Vanderplank occur north of 8 degrees 20'N in the north-west of Liberia. PMID- 2979552 TI - Light and suction trap catches of Culicoides midges in southern England. AB - 1. The seasonal flight periods are given for eleven species or species groups of Culicoides midges at Pirbright, Surrey, U.K., from 1979 to 1981 and in 1983. 2. Most species were relatively uniformly distributed both within the study site and between years; only C.clintoni Boorman was limited in its spatial distribution and only C.segnis Campbell and Pelham-Clinton varied greatly in numbers between years. 3. Species fall into one of three patterns of seasonal abundance: spring emergers, summer emergers, or spring to autumn emergers. 4. Only the C.obsoletus group showed a pattern of emergence that could correspond to distinct generations, of one or more species in the group. Three distinct emergences were observed, and possible explanations for these discussed. 5. For all species except C.impunctatus Goetghebuer the catches from suction/light and suction only traps are poorly correlated whereas for C.impunctatus they are highly correlated. This may suggest that this species is not greatly attracted to light. PMID- 2979553 TI - An experimental assessment of the tampan tick Ornithodoros moubata as vector of hepatitis B virus. AB - Wild-caught and colonized tampan ticks, Ornithodoros moubata (Murray), were fed on hepatitis B virus (HBV)-positive blood-means in a series of four experiments. Hepatitis B surface antigen (HBsAg) persisted in nymphal and adult ticks for up to 779 days, while the epsmark antigen (HBeAg) persisted in mature nymphs up to 13 days, in adult males up to 11 days and in adult females up to 16 days. HBsAg was transmitted trans-stadially through two moults during the life cycle but transovarial transmission did not occur. The surface antigen was transmitted by two out of fifteen single ticks into 0.4 ml aliquots of HBV-negative blood, although six groups of ticks failed to transmit into 5.5 ml aliquots of blood: this antigen was not transmitted to hamsters. HBsAg was detected in samples of the ticks' coxal and rectal fluid secretions always at the infecting feed and usually at the second feed. HBeAg was only detected in one of two samples of coxal fluid collected at the infecting feed. The results as a whole indicate that no biological multiplication of virus occurs in O.moubata but that mechanical transmission from ticks to man could occur by: (i) contamination of a person when crushing infected ticks; (ii) infection by bite; (iii) contamination with coxal fluid, especially by scratching bites. This is thought to take place among the Kavango tribe in their village huts in north-eastern Namibia where infestations of infected O.moubata occur. PMID- 2979554 TI - Fecundity and size in the housefly: investigations of some environmental sources and genetic correlates of variation. AB - Environmental sources and genetic correlates of variation in fecundity and size were studied in field and laboratory housefly populations in central Iowa, U.S.A. Electrophoresis of enzymatic proteins was used to monitor genotypes and heterozygosity. In the field, mean fecundity declined as adult densities increased. Fecundity varied greatly within breeding seasons. Fly body size varied spatially and temporally during summer and winter. Fecundity and size were positively correlated. Size was independent of genotypes at the six loci studied. No relationship was observed between heterozygosity and the magnitude or variability of body size. PMID- 2979555 TI - Leishmaniasis in Brazil. XXV. Sandfly vectors of Leishmania in Para State, Brazil. AB - Leishmania of the braziliensis complex were isolated from various members of the squamiventris series sandflies, including Psychodopygus chagasi (Costa Lima), P.s.maripaensis (Ready et al.), P.s.squamiventris (Lutz & Neiva), and also P.ayrozai (Barreto & Coutinho) and Lutzomyia umbratilis Ward & Fraiha. Three different serodemes of Le.b.guyanensis Floch were isolated from Lu.anduzei Rozeboom, Lu.umbratilis and Lu.whitmani Antunes & Coutinho, simultaneously captured in the same area. Unidentified Leishmania were isolated from P.claustrei Abonnenc et al., and trypanosomes from Lu.pinottii Damasceno & Arouck and an unidentified species of Lutzomyia. A naturally infected female P.s.maripaensis transmitted a braziliensis complex Leishmania, by bite, to a hamster. PMID- 2979557 TI - Host immunoglobulin G titre and antibody activity in haemolymph of the tick, Ornithodoros moubata. AB - Immunoglobulin G (IgG) in tick haemolymph was analysed immunochemically and biochemically for its antigenicity, antibody activity and relative concentration in a soft tick, Ornithodoros moubata (Murray) sensu Walton 1962 (Acari: Argasidae). Ouchterlony immunodiffusion tests showed that haemolymph from a tick engorged on rabbit IgG (or human IgG) through an artificial membrane, reacted with anti-rabbit IgG (anti-human IgG) but not with anti-human IgG (anti-rabbit IgG). This indicates that haemolymph of the fed tick contains IgG with a similar antigen specificity to host blood IgG. IgG from tick haemolymph was demonstrated by enzyme immunoassay to have the same antibody activity as ingested IgG. The IgG concentration in tick haemolymph was measured by a quantitative single immunodiffusion test. Changes of IgG titre after a bloodmeal were correlated with IgG activity, which was low for 5 days after a bloodmeal and then suddenly increased. The IgG titre reached a maximum 7 days post-engorgement, and remained high for over 4 months during and after oviposition. 125I-labelled IgG was injected into the tick haemocoel to determine the persistence of IgG in the haemolymph. Recovery of labelled IgG was low at 1 and 3 days, and high at 5, 8 and 16 days after engorgement. The data suggest that IgG in haemolymph disappears quickly soon after engorgement possibly by degradation and/or absorption (adhesion to tissues). PMID- 2979556 TI - The influence of permethrin-impregnated bednets and mass drug administration on the incidence of Plasmodium falciparum malaria in children in Sabah, Malaysia. AB - A small-scale trial was carried out in the Upper Kinabatangan district of Sabah, Malaysia, to determine the effect of using permethrin-impregnated bednets on malaria transmission. A total of 306 nylon bednets with cotton borders, impregnated at a dose estimated to have been 0.062 g permethrin/m2 of nylon netting, were distributed to 139 households in five villages. At the time of distributing bednets, mass drug administration with Fansidar plus primaquine was also administered to the human population to clear all parasitaemias due to Plasmodium falciparum Welch. In another village, for comparison, mass drug administration was the only intervention. After intervention measures in December 1984 and January 1985, the parasite rates in children declined in all villages during the first month, significantly more in the villages with impregnated bednets than in the control, thus proving that the nets had an impact on malaria. However, after about 2 months, parasite rates started to increase again. After 4 6 months, parasite rates in the villages with bednets approached the rate in the control village without nets. The increase in parasite rates was paralleled by a significant deterioration in the quality, physical condition and the degree of non-utilization of bednets. Entomological evaluation proved the efficacy of permethrin-impregnated nets for controlling Anopheles balabacensis Baisas and other anophelines. Bioassays (1 h exposure) of permethrin-impregnated bednets gave 100% mortality initially and 44-61% mortality after 85-106 days. Mosquito collections in treated bednets were significantly reduced for at least 217 days. The project failed to achieve prolonged suppression of malaria transmission for a combination of entomological, sociological and practical reasons which are discussed in relation to the objectives and implementation of future bednet studies. PMID- 2979558 TI - The life expectancy of phlebotomine sandflies: first field estimates from southern France. AB - 1. A field study of Phlebotomus ariasi Tonnoir, the vector of Leishmania infantum Nicolle in southern France, addressed the following questions: Is it possible to estimate reliably the life expectancy of this sandfly; can spatial or temporal variation in the life expectancy be detected, and is such variation significant for disease transmission? 2. Life expectancy was estimated by examining follicular relics in the ovaries of more than ten thousand females caught in light traps at seven sites in the Cevennes and the Garrigues, throughout their active period in 1985 and 1986. Whilst the distinction between nulliparous and parous flies was easily made, assessments of the number of times a parous fly had laid eggs were unreliable. Best estimates of life expectancy were therefore calculated from the parous rate. 3. Large samples collected from one site in the Cevennes in both years gave very similar estimates of life expectancy. 4. There was also no significant difference between estimates obtained from the Cevennes and the Garrigues, despite their distinct vegetation and climates. Therefore, large regional differences in sandfly population size and the prevalence of canine leishmaniasis cannot be explained by a difference in adult survival rate. 5. With no systematic annual or regional variation, a useful mean life expectancy can be calculated from the data collected at all sites in both years. It is 1.54 (SE 0.04) ovarian cycles. However, this estimate is sensitive to the assumption that survival rate is a discrete rather than a continuous variable. 6. Local variation in the parous rate may be associated with the proximity of traps to P.ariasi emergence sites. PMID- 2979560 TI - A record of trypanosomes from Ixodes ricinus in Britain. PMID- 2979559 TI - Honeydew sugars in wild-caught Phlebotomus ariasi detected by high performance liquid chromatography (HPLC) and gas chromatography (GC). AB - Phlebotomus ariasi Tonnoir sandflies were caught in light traps hung in oak trees and in a house in the Cevennes focus of leishmaniasis in the South of France. The flies were cryopreserved either immediately on removal from the traps, or after starvation for 6-7 days, or after 6-7 days starvation followed by exposure to oak infested with the aphid genera Lachnus or Thelaxes. After transportation to the laboratory, the sandflies were thawed and aqueous extracts of the crushed flies were analysed for their carbohydrate content using high performance liquid chromatography (HPLC) and gas chromatography (GC). Starved female sandflies lacked significant amounts of any saccharides. Four types of sugar, melezitose and its hydrolysis products turanose, glucose and fructose, were observed in flies which had been starved previously and then placed with Lachnus infested oak. The results also indicate the presence of hydrolysis products of melezitose: (a) in flies previously starved and placed with Thelaxes infested oak, (b) in P.ariasi cryopreserved direct from the light traps hung in oak trees infested with Lachnus and Thelaxes, and (c) flies caught in a house. Unidentifiably small quantities of a trisaccharide were also detected in the latter groups of flies. In previous tests, sugars were detected in P.ariasi after their exposure to aphid infested oak (Quercus ilex L.), but not when P.ariasi females were exposed to washed oak leaves without aphids. The results indicate that P.ariasi feed on melezitose and/or turanose, the main local source of which is aphid honeydew. A better understanding of sugar meal sources of sandflies using HPLC and GC techniques will assist in our understanding of sandfly/Leishmania relationships, parasite transmission and epidemiology. PMID- 2979561 TI - Sickness absenteeism as a test for the population's health status. PMID- 2979563 TI - Heat stress and voluntary dehydration in steelworkers. PMID- 2979562 TI - Some risk factors of coronary heart disease in miners of a new coal mining area. PMID- 2979564 TI - Allergy to chromium, cobalt and nickel in metal and electrical industry workers. PMID- 2979565 TI - The biochemical indices of genetic susceptibility to cancer. Acetylation phenotype and bladder cancer. PMID- 2979566 TI - Oxidative phenotype as a marker of susceptibility to larynx carcinoma. PMID- 2979567 TI - In vivo study on the frequency of sister chromatid exchange (SCE) and micronuclei (MN) induced by cyclophosphamide in bone marrow and spleen cells in rodents. PMID- 2979568 TI - Influence of a static magnetic field on the reproductive function, certain biochemical indices and behaviour of rats. PMID- 2979570 TI - Major hazard control. PMID- 2979569 TI - Radiation hazard in Polish spas--ten years' experience and studies. PMID- 2979571 TI - Entrainment of split circadian activity rhythms in hamsters. AB - Hamsters that showed splitting of their circadian rhythms of wheel-running activity following long-term exposure to constant illumination (LL) were exposed to light-dark (LD) cycles with 2-hr dark segments, and with periods of 24.00, 24.23 or 24.72 hr. For comparison, hamsters showing nonsplit rhythms were also studied. In all cases of split rhythms, at least one of the two split components entrained to the LD cycles. In some animals, the second component continued to free-run until it merged with the entrained component, while in others, the second component also entrained to the LD cycle but maintained a stable phase angle of 6-14.5 hr relative to dark onset. These results were obtained in cases where the period of the LD cycle was shorter than that of the split rhythms and in cases where it was longer, implying that split components can be phase advanced as well as phase-delayed by 2 hr of darkness. Three hamsters that showed stable entrainment of split rhythms were allowed to free-run in LL. The LD cycles were then reinstated, but instead of overlapping with the first component, as it did before, the dark segment was timed to overlap with the second. The entrainment patterns that ensued were similar to the ones obtained during the first LD exposure, indicating that the two split components respond to darkness in a qualitatively similar fashion. These results are further evidence that the pacemaker system underlying split circadian activity rhythms in hamsters is composed of two mutually coupled populations of oscillators that have similar properties, including a bidirectional phase response curve. Such a dual oscillator organization may also underlie normal, or nonsplit, activity rhythms, as suggested by Pittendrigh and Daan (1976c), but the data are also compatible with the alternative view that the circadian pacemaker consists of a large number of coupled oscillators, which only dissociate into two separate populations in some animals under conditions of moderate LL intensity. PMID- 2979572 TI - Circannual rhythms of ground squirrels: role of the hypothalamic paraventricular nucleus. AB - Female golden-mantled ground squirrels, maintained in an LD 14:10 photoperiod at 23 degrees C, sustained lesions of the paraventricular nucleus (PVN) or sham operations. Body weight and reproductive status were recorded weekly pre- and postoperatively. Bilateral lesions of the PVN did not eliminate, phase-shift, or otherwise disrupt the circannual rhythms of body mass or reproduction. Absolute levels of body weight were unaffected by PVN ablation. The PVN is not an essential component of the oscillatory system that generates circannual cycles in ground squirrels. PMID- 2979573 TI - Circadian rhythms in the electroretinogram of the cockroach. AB - Circadian regulation of the amplitude of the electroretinogram (ERG) of the cockroach Leucophaea maderae was investigated. Two components of the ERG exhibited circadian rhythms in amplitude. Interestingly, the peak amplitudes for the two rhythms were approximately 12 hr out of phase. The dominant corneal negative potential (the "sustained component") exhibited maximum amplitude during the subjective night. A second corneal negative potential (the "off-transient") was at a maximum during the subjective day. Intensity-response curves of the sustained component were measured at both the peak and trough of the rhythm. The results showed that the circadian rhythm in amplitude reflected a sensitivity change equivalent to 0.2-0.6 log unit of intensity. An effort was also made to identify the anatomical locus of the pacemaking oscillator for the ERG rhythm in a series of lesion experiments. Neural isolation of the optic lobe from the midbrain by bisection of the optic lobe proximal to the distal edge of the lobula had no effect on the circadian rhythm of ERG amplitude. Bisection of the optic lobe distal to the lobula abolished the ERG amplitude rhythm. These results suggest that the pacemaker is located in the optic lobe near the lobula; that its motion continues in the absence of neural connections with the rest of the nervous system; and that its regulation of ERG amplitude depends on neural pathways in the optic lobe. PMID- 2979575 TI - Analysis of mutual circadian pacemaker coupling between the two eyes of Bulla. AB - The eyes of Bulla, a marine snail, express a circadian rhythm in the frequency of optic nerve compound action potentials (CAPs). The two ocular pacemakers are mutually coupled, and their interaction can be observed in vitro. The evidence for mutual coupling, as demonstrated in the present experiments, was as follows: (1) When intact Bulla were placed into darkness for up to 72 days, the two pacemakers did not desynchronize. (2) The free-running period of the ocular rhythm in the intact system (24.4 hr) was longer than the free-running period of the rhythm recorded from isolated eyes (23.7 hr). (3) When the two ocular pacemakers were experimentally desynchronized in vitro, resynchronization occurred if the pacemakers were allowed to interact for 48 hr. The coupling signals are most likely the CAPs. These impulses are conducted through the central ganglia and emerge as efferent impulses in the opposite optic nerve. Ocular-derived efferent impulse activity affects spontaneous impulse production in the target eye and alters the waveform of the circadian rhythm. The coupling pathway mediating syncrhonization consists of the two optic nerves, the cerebral ganglia, and the cerebral commissure. The demonstration of coupling in vitro provides a new opportunity for studying the cellular mechanisms underlying mutual pacemaker entrainment. PMID- 2979574 TI - Circadian regulation of pineal melatonin and reproduction in the Djungarian hamster. AB - Gonadal state, pineal melatonin rhythms, and locomotor activity rhythms were examined in juvenile male Djungarian hamsters exposed to non-24-hr light cycles ("T-cycles") or to full photoperiods. At the end of 1 month, hamsters exposed to a 1-hr pulse of light every 24.33 hr (T 24.33) exhibited small testes, whereas those receiving the same amount of light every 24.78 hr (T 24.78) displayed stimulated gonads, ten-fold larger in size. Accompanying the nonstimulatory effect of the T 24.33 cycle were nocturnal peaks in both pineal melatonin content and serum melatonin concentration which were longer by approximately 4 hr than those observed on the photostimulatory T 24.78 cycle. Exposure to an intermediate length T-cycle (T 24.53) resulted in a mixed gonadal response and in pineal and serum melatonin peaks of intermediate duration. Wheel-running activity was entrained to the T-cycles such that light was present only near the beginning of the subjective night, its phase (relative to activity onset) differing only slightly among T-cycle groups. Hence the durational differences observed in the melatonin peaks were apparently not due to the acute suppressive or phase advancing effects of morning light on melatonin biosynthesis, but were rather the result of differences in the endogenous control of pineal activity by the circadian pacemaker system. While no strong correlation was detected between gonadal state and the phase of locomotor activity onset relative to the light pulse, a significant correlation was observed between gonadal state and the duration of daily locomotor activity (alpha). These data were compared to similar measures obtained from hamsters exposed to long-versus short-day full photoperiods (LD 16:8 vs. LD 10:14). In summary, the results of this study indicate involvement of the circadian pacemaker system of Djungarian hamsters in the control of pineal melatonin synthesis and secretion, and in photoperiodic time measurement. Furthermore, these data strengthen the hypothesis that it is the duration of nocturnal pineal melatonin secretion that is the critical feature of this neuroendocrine gland's photoperiodic signal. PMID- 2979576 TI - Development of hamster circadian rhythms: prenatal entrainment of the pacemaker. AB - Individual hamster pups were maintained in constant dim light from just prior to birth, and their circadian wheel-running activity rhythms were recorded beginning at 18 days of age. Records of the postweaning free-running activity rhythm were used to determine the phase of a pup's rhythm on the day of weaning. Two groups of pups (LD and DL) were born to mothers that had been entrained before birth to light-dark cycles 12 hr out of phase. Both groups of pups were raised in constant dim light by foster mothers that had been entrained to only one of the prenatal cycles (LD). Thus pups born to mothers from different cycles were exposed to identical rhythmic environments postnatally. Despite the similar postnatal treatment, the two groups of pups showed activity rhythms at weaning with very different phases. The LD pups, born to and raised by LD mothers, were approximately synchronous with one another and with their foster mothers. The DL pups, born to DL mothers, but raised by LD mothers, were not synchronous with one another or with their foster mothers. Half of the DL pups showed phases predicted by their prenatal treatment, but the other half showed scattered phases. The results demonstrate that phase at weaning is affected by prenatal rhythmicity, and suggest that the circadian pacemaker underlying the activity rhythm is already functional and entrained at, or before, birth. PMID- 2979577 TI - Genetic correlation between circadian eclosion rhythm and photoperiodic diapause in Drosophila littoralis. AB - Populations of Drosophila littoralis are known to be latitudinally highly variable in photoperiodic adult diapause and pupal eclosion rhythm. Phenotypic correlations between the two time-measuring systems among the strains from different latitudes are, however, weak. In the present study, two differing strains were crossed reciprocally in order to search for causal (genetic) correlations between the two traits in the strains. Segregation in the F2 generations showed that variation in each trait was based on a few variable loci only. In the F2, flies having different eclosion times also differed in their diapause. This association was not complete and could have been due to genetic linkage between the traits. For that reason, the hybrid generations were raised for eight generations more to allow recombination between the traits. In F8, selection against diapause was started in the lines by raising them in a light dark cycle of 15:9, where only females of the southern type reproduce. After eight selected generations, the lines were studied for the traits. Diapause was completely of the southern type, and the eclosion rhythm had also changed in parallel. The change in the phase of the free-running rhythm was not complete. From the present experiment, and from earlier knowledge of the geographical variation in D. littoralis, I conclude that the same pacemaker that is seen in the eclosion rhythm could also participate in daylength measurement for diapause. However, there are also noncorrelated variable parts in the measuring systems of both traits, which may mask the correlated variation. PMID- 2979578 TI - Masking of the circadian rhythms of heart rate and core temperature by the rest activity cycle in man. AB - Heart rate and core temperature are elevated by physical activity and reduced during rest and/or sleep. These masking effects may confound interpretation of rhythm waveforms, particularly in situations where the rest-activity rhythm has a different period from that of the core temperature rhythm. Such desynchronization often occurs temporarily as an individual adjusts to a new work shift or to a new time zone following rapid transmeridian travel, making it difficult to assess the impact of such schedule changes on the circadian system. The present experiments were designed to estimate the magnitude of these masking effects, by monitoring the heart rate, rectal temperature, and nondominant wrist activity (2-min samples) of 12 male subjects during 6 days of normal routine outside the lab and during 6 days of strict bedrest. Subjects also kept sleep, dietary, and exercise logs throughout the study. Average (20-min) waveforms were computed for each subject and each rhythm, at home and in bedrest. In addition, data were partitioned according to self-reported sleep and wake times and were analyzed separately for each state. Average waveform comparisons indicated that about 45% of the range of the circadian heart rate rhythm during normal routine was attributable to the masking effects of activity during wake, which also produced a 16% elevation in mean heart rate during wake and an 11% increase in mean heart rate overall. (Analysis of variance indicated that mean heart rate during sleep at home was not significantly different from the mean during sleep in bedrest.) On average, about 14% of the range of the circadian temperature rhythm during normal routine was attributable to the effects of activity masking. However, the change in range of the temperature rhythm, from home to bedrest, was very variable between subjects (-41% to +13%). This variability was not accounted for by age or by reported frequency of exercise at home. Normal activity during wake increased the mean temperature during wake by an average of 0.16 degrees C and the overall mean by about 0.12 degrees C. (Analysis of variance indicated that mean temperature during sleep at home was not significantly different from the mean during sleep in bedrest.) A 10-hr "night" (lights-off from 2200 to 0800 hr) was provided during bedrest, within which subjects could select their own sleep times. Times of sleep onset and wake onset were not significantly different between home and bedrest.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2979579 TI - Photoperiodic control of diapause induction and termination in Ostrinia nubilalis: two different clocks? AB - Both diapause induction and diapause termination are under photoperiodic control in the lepidopteran, Ostrinia nubilalis. In the present study, induction of diapause was maximal in light-dark (LD) cycles that contained 12 hr of light alternating with 12 hr of darkness (LD 12:12). Termination of diapause was maximal in LD 16:8. Diapause termination also occurred rapidly in non-24-hr LD cycles that possessed an 8-hr dark phase. In each of these cases, the period of the LD cycle was not important. Diapause termination did not, however, occur rapidly in non-24-hr LD cycles that lacked an 8-hr dark phase. Thus, the clock mechanism underlying the termination response resembles an hourglass in its behavior. This is in contrast with what is known about induction of diapause. Here it has been demonstrated that the circadian system is somehow involved. It is thus possible that two different physiological clocks underlie these responses. PMID- 2979580 TI - Photoperiodism in Ostrinia nubilalis: a new protocol for the analysis of the role of the circadian system. AB - A 12-hr dark period, at a temperature high enough to permit time measurement to occur, is necessary for maximal induction of larval diapause in the European corn borer, Ostrinia nubilalis. In the present study, induction of diapause only occurred in a periodic environment. This was in the form of certain (1) light dark (LD) cycles at a constant temperature; (2) thermoperiods in constant darkness (DD), but not constant illumination (LL); and (3) LD cycles with concurrent thermoperiods. A light-break experiment protocol, in which the pulses systematically scan the cold and warm phases of a thermoperiod in DD, is discussed as a way of helping clarify how seasonal time measurement is effected in Ostrinia. PMID- 2979581 TI - Meal timing in humans during isolation without time cues. AB - In an underground isolation unit, 46 subjects lived singly without time cues under conditions of self-selected light-dark (LDs) cycles or constant illumination (LL). They all developed free-running circadian rhythms that either remained internally synchronized (i.e., with equal periods in all functions; n = 24) or became desynchronized by a sudden lengthening (n = 14) or shortening (n = 8) of the sleep-wake cycle. Six further subjects were synchronized to 24 hr by an externally controlled LD cycle. All subjects had to prepare their own meals. Signals were given by the subjects when they woke up, had a meal, and retired. Of the 52 subjects, 43 consistently had three meals per "day," and 9 had two meals, irrespective of the length of the circadian cycle and despite the fact that, due to desynchronization, wakefulness varied from over 30 hr to less than 12 hr. The intervals between meals, between wake-up time and breakfast, and between last meal and bedtime were "stretched" or "compressed" in strong proportionality to the duration of wakefulness. PMID- 2979582 TI - Circadian activity rhythms in squirrel monkeys: entrainment by temperature cycles. AB - Eight squirrel monkeys (six male, two female), kept singly in wire-mesh cages, were housed in groups of three in a climatic chamber. Locomotor activity was recorded by means of photocell systems or mobile climbing trees. For time spans of several weeks, the animals were alternately exposed to each of three different conditions: (1) constant conditions at various levels of light intensity and ambient temperature; (2) a 24-hr light-dark (LD) cycle at constant temperature; (3) a 24-hr cycle of low (about 17 degrees C) and high (32 degrees C) temperature in constant illumination (LL). In constant conditions, the free-running activity rhythms always had periods longer than 24 hr, without systematic dependencies on ambient temperature, but with a tendency to lengthen the period when the intensity of illumination was increased. All animals were entrained by a weak LD zeitgeber (10:1 lux). Exposure to cycling ambient temperature resulted in entrainment in about 45% of all tests. Usually, the entrained animals were active during the cold fraction of the cycle, but some records suggested entrainment in a warm-active mode. In addition, temperature changes exerted strong masking effects, mainly with increases of activity during the warm fraction of the cycle. PMID- 2979583 TI - Circadian photoentrainment: parameters of phase delaying. AB - Experiments were carried out using simulated den cages to delineate specific characteristics of phase delaying in circadian photoentrainment of a nocturnal rodent, the flying squirrel. The principal experiments entailed presentation of one to five consecutive 15-min white-light pulses per activity cycle at activity onset to animals free-running in darkness, in order to determine the immediate and final phase-shifting effect. Auxiliary experiments recorded entrainment patterns on light-dark (LD) schedules in the den cages. Phase response curves (PRCs) based on 15-min white-light pulses in standard wheel cages were also constructed for these animals as background information for interpreting the phase-delaying experiments. Exposure of a den animal to light by light sampling at the time of initial arousal from the rest state at circadian time (CT) 12, either by an LD schedule or by a 15-min light pulse, resulted in a return to the nest box for a short rest period. The phase delay occurring after a single light exposure at activity onset was equal to the induced rest, thus suggesting an immediate phase shift. The maximum delay was about 1 1/2 hr/cycle, with the amount of delay related to the number of light exposures. During the photoentrained state on an LD schedule, the activity rhythm of a den-housed animal was essentially free-running on the days following a phase delay. The data are used to expand current models for photoentrainment of circadian activity rhythms in nocturnal rodents. PMID- 2979584 TI - Cellular analysis of ocular circadian pacemaker coupling in Bulla: role of efferent impulses in phase shifting. AB - The eyes of Bulla gouldiana, a marine snail, contain circadian oscillators that are coupled to each other. Obvious candidates for the coupling signals are the optic nerve compound action potentials (CAPs) that express the circadian rhythm and lead to efferent impulses in the contralateral optic nerve. In the present experiments, the role of the CAPs as coupling signals was evaluated. We found that, following desynchronization of the two ocular oscillators by phase-delaying one eye with manganese, subsequent phase shifts in the initially unshifted ocular rhythm only occurred during the time that efferent optic nerve signals were present. In addition, in the absence of ocular desynchrony, phase shifts of the ocular rhythm could still be effected by activation of the efferent pathway. The influence of efferent impulses on identified retinal cells was also evaluated. No effect of efferent signals on receptor layer cells was detected, while it was found that efferent impulses generated depolarizations in basal retinal neurons (BRNs), the putative circadian oscillator cells. Depolarization of the BRNs has been shown previously to be involved in the light entrainment pathway. Depolarization appears to be similarly involved in the coupling pathway, since membrane depolarizations that mimicked the efferent-induced postsynaptic potentials likewise generated phase shifts of the ocular rhythm. PMID- 2979585 TI - Dose-dependent entrainment of rat circadian rhythms by daily injection of melatonin. AB - Previous work in our laboratory has shown that daily injection of large doses of the pineal hormone melatonin entrains the free-running locomotor rhythms of rats held in constant darkness and synchronizes the disrupted patterns of rats maintained in constant bright light. The present experiments determined the dose response characteristics of entrainment to daily melatonin injections and made preliminary biochemical estimates of blood melatonin levels and half-lives after two critical doses of the hormone. The data indicated that the median effective dose for melatonin as an entraining agent in free-running rats was 5.45 +/- 1.33 micrograms/kg, considerably lower than doses previously employed and lower than doses employed in reproductive and metabolic studies in rats and hamsters. The data further indicated that the response to melatonin was quantal; rats either entrained to melatonin or they did not. No "partial entrainment" was evident, nor were there differences in phase angle, activity, or period among all effective doses. Biochemical estimates of blood melatonin after either 1 mg/kg or 1 microgram/kg of melatonin indicated that all effective doses resulted in supraphysiological levels of blood melatonin, although doses of 1 microgram/kg resulted in blood levels that were within one order of magnitude of normal nighttime values. Together, the data suggest that the rat circadian system is sensitive to the pineal hormone melatonin at or below doses required to effect rodent reproduction. Whether this sensitivity reflects a role for the pineal gland in rat circadian organization, however, still remains to be determined. PMID- 2979586 TI - Circadian participation in the photoregulation of testis activity and prolactin secretion in the mink, a short-day breeder. AB - It has been demonstrated that an endogenous mechanism is involved in photoperiodic time measurement in the mink, a short-day-breeding mannal. A study of testicular activity (testicular volume, plasma testosterone concentration) and plasma prolactin level was carried out in sexually resting minks (the experiment began in November). Groups of minks were kept in the natural photoperiod or subjected to different resonance light-dark (LD) cycles (LD 4:8, LD 4:20, LD 4:32, LD 4:44); an additional group of animals was reared in an ahemeral photoperiod (LD 4:16). A rapid increase of testicular activity was observed in control animals or those kept in LD 4:20 (T 24) and LD 4:44 (T 48). In the other groups of animals, those kept in LD 4:8 (T 12), LD 4:32 (T 36), and LD 4:16 (T 20), testicular function remained at rest. Prolactin secretion was, in contrast, stimulated in the groups kept in LD 4:8 (T 12). LD 4:32 (T 36), and LD 4:16 (T 20), and remained low in the groups kept in LD 4:20 (T 24) and LD 4:44 (T 48). These results show that the effects of the different photoperiodic regimens do not depend on the duration of the photophase, but rather on the period of the LD cycles. The LD cycles that allow an increase of testicular function are those that are inhibitory to reproduction in birds and long-day-breeding mammals. To explain these results, it is suggested that in the mink exposure to light during the circadian photosensitive phase induces inhibition of testicular activity and stimulation of prolactin secretion. To explain the opposite effects of a single photoperiod on testicular function and secretion of prolactin, the hypothesis has been advanced that, in the mink, long days might simultaneously inhibit hypothalamic luteinizing-hormone-releasing hormone (LH-RH) activity and prolactin inhibiting factor (PIF) activity. PMID- 2979587 TI - Insect photoperiodism: diversity of results in night-break experiments, including nonresponsiveness to light. AB - Three night-break experiment protocols were utilized in an attempt to help clarify the role of the circadian system in photoperiodic time measurement in the European corn borer, Ostrinia nubilalis. Larvae raised in a light-dark (LD) cycle consisting of 12 hr of light alternating with 12 hr of darkness (LD 12:12), at a constant temperature of 30 degrees C, enter a state of arrested growth and development known as diapause (Takeda and Skopik, 1985). In the present research (Experiment 1), the induction of diapause was prevented by 1-hr light pulses that systematically scanned the dark phase of LD 12:12. Thus, the importance of 12 hr of uninterrupted darkness for maximal induction of diapause is stressed. The same experimental protocol applied to larvae already in diapause (Experiment 2), however, resulted in a bimodal curve of diapause termination. Although this result is consistent with the proposition that a nonperiodic hourglass timer underlies this event (Skopik and Takeda, 1986), it does not rule out the circadian system. Like LD 12:12, a thermoperiod in constant darkness (12 hr at 4 degrees C alternating with 12 hr at 25 degrees C) also induces diapause. Scanning such a thermoperiod with 1-hr light pulses, however, resulted in only a small effect (reduction of diapause) when light fell in the early to middle part of the warm phase (Experiment 3). Thus, the time-measuring system, under these experimental conditions, showed only a weak response to light. This unexpected result is discussed with respect to Experiment 1 and two general models that have been proposed to account for photoperiodic time measurement in insects. PMID- 2979588 TI - Acquisition of circadian bioluminescence data in Gonyaulax and an effect of the measurement procedure on the period of the rhythm. AB - During measurements of the circadian (approximately 24-hr) rhythms of spontaneous bioluminescence in the marine dinoflagellate Gonyaulax polyedra, the individual cultures in vials were shielded from otherwise constant dim light for 1-3 min every 20-60 min by a photomultiplier housing that was moved from vial to vial. The high-frequency dark pulses caused a small but consistent shortening of the free-running circadian period, but there was no indication that the dark pulses caused entrainment. Hardware and software components of the microcomputer controlled data collection system are described. A microcomputer controlled the movement of the photomultiplier and acquired the data via an analog-to-digital converter. The algorithms distinguished and separately recorded background glow, intermittent flashes, and total light from populations ranging in number from 10(3) to 10(5) cells in volumes from 1 to 10 ml. Fast video display techniques allowed continuous on-line viewing of incoming data, together with a display of the data recorded over the preceding day or two. Detection of mechanical and software errors coupled with recovery systems maintained high reliability of data collection. PMID- 2979589 TI - Effects of feeding cycles on circadian rhythms in squirrel monkeys. AB - Squirrel monkeys (Saimiri sciureus) were housed singly in cages equipped with a tree for climbing to measure locomotor activity, and with a movable food cup that could be arrested automatically. The animals were kept in continuous dim illumination (LL), twice interrupted by several weeks of entrainment by a light dark (LD) 12:12 cycle. Apart from three control sections in which the food cups were unlocked continuously (ad libitum feeding), food was accessible for 3 hr per day only, with interfeeding intervals varying from 23 to 26 hr (periodic restricted feeding, or RF). During LD entrainment, the imposition of an RF schedule resulted in anticipatory behaviors, represented by increased tugs at the food cup and a pause in locomotor activity preceding the feeding time. In LL, the animals showed free-running circadian rhythms of locomotor and "feeding" activity that nearly always persisted when ad libitum feeding was replaced by RF. The period (tau) of the free-running rhythm was slightly modulated in relation to the varying interfeeding intervals (T), but entrainment was never achieved except in one test with an animal whose tau was very close to T. It is concluded that periodic availability of food represents an extremely weak zeitgeber, if any, for the circadian pacemaker of squirrel monkeys. PMID- 2979590 TI - Circannual rhythms of ground squirrels: a test of the frequency demultiplication hypothesis. AB - The frequency demultiplication hypothesis (FDH) posits that circannual rhythms are generated from circadian cycles by frequency transformation to the lower frequency rhythm. To test the FDH, we determined the periods of the circannual body mass and estrous cycles of golden-mantled ground squirrels with circadian locomotor activity rhythms entrained to 23-, 24-, or 25-hr days (T-cycles). Circannual period length did not differ among squirrels entrained to the different T-cycles; intergroup ranges were 298-314 days and 303-312 days, respectively, for body mass and estrus. These results are not consistent with the FDH and suggest instead that separate mechanisms generate circadian and circannual rhythms. In ground squirrels the circannual system influences circadian organization, but a reciprocal influence of circadian on circannual rhythms has yet to be demonstrated. PMID- 2979591 TI - Photoperiodic and pineal effects on food intake, food retrieval, and body weight in female Syrian hamsters. AB - Food intake, food retrieval, and body weight changes were measured in female Syrian hamsters maintained under long and short photoperiods, and in animals sustaining pinealectomy or sham surgery. Animals maintained under short photoperiods gained more weight, ingested more food, and brought more food from more distant compartments into their nest compartments than did animals maintained under long photoperiods. Pinealectomy prevented the short-day elevations in body weight and food retrieval during the period of gonadal recrudescence, but it did not produce significant changes during the period of gonadal regression; in contrast, pinealectomy prevented the short-day increases in food intake only during initial weeks of the period of gonadal regression: The effects of photoperiod or pinealectomy were not evident for any of the dependent measures during pregnancy. The role of the pineal gland in mediating short-day effects is discussed. PMID- 2979592 TI - Slow-wave sleep in daytime and nocturnal sleep: an estimate of the time course of "Process S". AB - Daan et al. (1984) have proposed that sleep and wakefulness are regulated, in part, by a "Process S" that increases during wakefulness and declines during sleep. Data derived from several studies were taken to determine the time course of Process S during both wakefulness and sleep. As required by the model, slow wave-sleep (SWS; an index of Process S) was found to increase exponentially as a function of prior wake time (equation 1) and to decline exponentially as a function of time asleep (equation 2). The equations accounted for 91% and 96% of the variance, respectively. In addition, equation 1 accurately predicted the amount the amount of SWS in the first hour of nocturnal sleep. PMID- 2979594 TI - Preventive toxicology: examining the coherence of the system in Poland. PMID- 2979595 TI - Theory and practice of establishing occupational exposure limits in Poland. PMID- 2979593 TI - Lesions of the thalamic intergeniculate leaflet alter hamster circadian rhythms. AB - We have investigated the effects of destruction of the geniculo-hypothalamic tract (GHT) on the circadian system of golden hamsters. In the first experiment, intact hamsters were housed in constant darkness, and phase shifts in running wheel activity rhythms were assessed following 15-min light pulses administered at circadian time (CT) 12 (defined as the beginning of activity), CT 14, CT 18, and CT 20. Responses to light pulses at the same CTs were then reassessed after GHT lesions. Hamsters with complete lesions showed decreases in phase advances caused by light pulses at CT 18 and CT 20. Phase delays elicited by light at CT 12 and CT 14 were not altered. In a second study, intact and GHT-ablated hamsters housed in constant light received 6-hr dark pulses at various CTs. Hamsters with complete GHT ablation showed smaller advances than controls to dark pulses centered on CT 8-10. After 110 days in constant light, 7 of 10 intact hamsters showed splitting of their activity rhythms into two components, while only 1 of the 8 similarly treated ablated hamsters displayed dissociated activity components. Ablated hamsters had significantly shorter free-running periods during the first 35 days of exposure to constant light than did the intact hamsters. These results demonstrate that destruction of the GHT in the hamster alters phase shifting in response to periods of light or dark, and they indicate a role for the GHT in mediating several photic effects on the circadian system. PMID- 2979596 TI - Metabolic interaction and neurological effect of combined exposure to acrylamide and ethanol. PMID- 2979597 TI - Protective effect of phenobarbital in liver injury by kerosene hydrocarbons in rats. PMID- 2979598 TI - The application of psychological methods for evaluation of effects of occupational exposure to neurotoxic substances. PMID- 2979599 TI - Dose-effect relationship theory based on metabolic kinetics of natural endogenous radioprotectors. PMID- 2979600 TI - Human organ specific autoimmunity: personal memories. PMID- 2979601 TI - Autoimmunity: a personal memoir. PMID- 2979602 TI - Perturbation of the autoimmune network. I. Immunization with anti-idiotypic antibodies prior to challenge with antigen induces quantitative variations in the autoantibody response. AB - The effect of immunization with anti-idiotypes on the production of autoantibodies subsequently induced with nominal antigen, thyroglobulin, was investigated in BALB/c mice with three distinct rabbit antibodies and one syngeneic monoclonal antibody (mAb) specific for an idiotype (Id62) borne on a mouse monoclonal autoantibody to thyroglobulin. In these in vivo experiments, dose, route and form of the anti-idiotypic immunization were kept constant, but the time interval between exposure to anti-idiotype and antigen challenge was varied. When the interval was short (two weeks), heterologous anti-Id62 antibodies predisposed to suppression of the autoantibody response in 2/3 instances. Suppression was also obtained in mice immunized with a syngeneic mAb directed against Id62. In each case, a prominent idiotype-positive (Id') response was measured in the serum. Adsorption/elution studies indicated that Id' molecules in suppressed mice were principally non antigen-binding. On the other hand, when the time interval between injections was long (14 weeks), mice preimmunized with one rabbit anti-Id62 antibody showed a markedly increased autoantibody response. The Id' component in enhanced mice was largely enriched for antibodies reacting with the antigen. These findings suggest that the state of activation of autoreactive clones and the type of anti-idiotypes used may be determinant factors in the overall effect of manipulation of autoimmune responses with anti-idiotypic antibodies. PMID- 2979603 TI - Long-acting thyroid stimulator: how receptor autoimmunity was discovered. PMID- 2979604 TI - Allospecific DR gene expression on the human thyroid cell. AB - Human thyroid epithelial cells, stimulated with recombinant gamma interferon (IF) (100 U/ml), were HLA-DR typed by means of antibody-dependent, complement-mediated cytotoxicity. The thyroid cell lysis reaction patterns were found to be similar to those obtained with autologous peripheral B cells in four of five separate experiments. Since the cytotoxic reactions with thyroid cells were sometimes incomplete with many antisera, a two-color fluorescence technique was developed to measure the specificity of the antibody-dependent, complement-mediated lysis. Using constitutively HLA-D antigen-positive Raji B lymphoblastoid cells as controls, we showed that non-polymorphic HLA-D and HLA-DR determinants could be detected on the thyroid cell surfaces by this cell lysis approach. In addition, gamma IF stimulated thyroid cells from a DR3 positive individual were lysed to a significantly greater than unstimulated autologous thyroid cells with the use of a specific monoclonal antibody to HLA-DR3, thus confirming the original HLA typing studies. These data demonstrate, for the first time, that the expression of HLA-DR antigen on the surface of human thyroid cells is allospecific and may, therefore, play an important role in the immunopathology of autoimmune thyroid disease. PMID- 2979605 TI - Polyclonal nature of islet cell antibodies in insulin-dependent diabetes. AB - Islet cell antibodies (ICA) are associated with insulin-dependent diabetes mellitus (IDD) and have been proposed as predictive markers for the disease. To determine whether ICA result from the activation of single autoreactive B lymphocyte clones or are the result of polyclonal B-cell activation, we assayed ICA using polyvalent antisera specific to kappa or lambda light chains as well as monoclonal antibodies to IgG1, IgG2, IgG3 and IgG4 heavy chains by indirect immunofluorescence. Sera from 38 newly diagnosed IDD patients with IgG-ICA titers greater than 1:8 by end-point dilution were studied. ICA of both kappa and lambda light chains were present in all sera. The ICA were predominantly of the IgG1 subclass (38/38), although ICA were also found to be IgG2 in 53% (20/38), IgG3 in 29% (11/38) and IgG4 in 16% (6/38). The distribution of IgG heavy chains in ICA was compared to the ICA titer, age of onset of IDD and HLA-DR phenotype of the patient. No statistical correlation could be detected at a P value less than 0.05. Our findings more likely exclude the occurrence of a single aberrant lymphocyte clone secreting ICA that may have arisen by somatic mutation in individual patients. Rather, these results are consistent with the hypothesis that ICA arise by polyclonal B-lymphocyte activation as a result of a defect of immune regulation. Since human antibodies to protein antigens are found predominantly in the IgG1 subclass, our findings support the belief that the autoantigen involved in the stimulation of ICA formation is comprised, at least in part, of protein. PMID- 2979606 TI - Comparison and standardization of measurement of anti-acetylcholine receptor antibody between laboratories. AB - The presence of autoantibodies to the acetylcholine receptor (anti-AChR) is useful in the diagnosis of myasthenia gravis, and their titre correlates with severity of the disease. Standardization of their measurement is therefore clinically important. Six laboratories world-wide were asked to determine anti AChR under local conditions in coded samples and to repeat the measurement on the same samples recorded. There was a high degree of consensus over rank order of the samples but a wide systematic variation in the titres obtained. Standardization of units is an important next step in improving the comparability of anti-AChR data between laboratories. PMID- 2979607 TI - An analysis of autoimmunity through studies of DNA antibody idiotypes. AB - The existence of idiotypic networks, first postulated over 12 years ago, is now widely recognised. Idiotypic analyses of autoantibodies have been reported among both hybridoma-derived and naturally occurring immunoglobulins. In this review the many studies of idiotypes detected on anti-DNA antibodies, notably one designated 16/6, are analysed to see what clues they offer to our understanding of autoimmunity. The links between infection and autoimmunity are emphasised by this analysis. It is also obvious that idiotypes first identified an autoantibodies are not confined to these immunoglobulins. Thus, the 16/6 idiotype originally described on a hybridoma-derived monoclonal anti-DNA antibody has also been identified on naturally occurring antibodies binding the Klebsiella polysaccharide K30. PMID- 2979608 TI - Origin of DNA from DNA/anti-DNA antibody immune complexes in systemic lupus erythematosus patients. PMID- 2979609 TI - Experimental autoimmune glomerulonephritis in chickens. II. Adoptive transfer of disease with mononuclear cells. AB - Autoimmune glomerulonephritis was induced in donor chickens by immunization with chicken, human, turkey or bovine glomeruli. Lymphocytes from the kidneys of each group and the spleen of donors immunized with chicken glomeruli were isolated and injected intravenously into six-week old syngeneic SC chickens. The disease was transferred to bursectomized/non-bursectomized recipients by cells from donors immunized with chicken and bovine glomeruli but not by cells from chickens immunized with human and turkey glomeruli. The transferred disease correlated with interstitial infiltrates in donors. No antibody was detected on the glomerular basement membrane or in the circulation of any recipients. Glomerulonephritis in this model can be transferred by specifically sensitized T lymphocytes alone without antibody. This new pathogenic process may play an important role in the development of glomerulonephritis in other animal models as well as in man, where minimal or no immune deposits are observed despite glomerulonephritis. PMID- 2979610 TI - Thymosin and the spontaneously diabetic BB rat. AB - The biological basis for autoimmunity and immunoincompetence in the BB rat has yet to be localized. In spite of normal thymic histology, thymocyte subsets and blastogenesis, thymus gland products (thymosins) have yet to be studied. In the present report, thymus gland function was studied by measuring thymosin alpha 1 levels at one time point in the BB rat compared with control rates, and BB rat responses to exogenous thymosin (Thymosin fraction 5) were observed. At five months of age, BB rats had thymosin alpha 1 levels comparable to Lewis and Wistar furth rats. Thymosin fraction 5 increased the ratio of peripheral blood W3/25 positive to OX8 positive cells, but otherwise had no effect on the BB rats' T cell immunodeficiency, or frequencies of tissue autoantibodies or insulin dependent diabetes. Although B-lymphocyte counts were normal in BB rats, splenocyte responses to B-lymphocyte mitogens were depressed. However, thymosin fraction 5 improved the BB rat B-lymphocyte blastogenesis to near normal for Mycoplasma neurolyticum. Coupled with our previous work, our results suggest that the immune derangement in the BB rat resides outside the thymus. PMID- 2979611 TI - Immunological studies of HTLV-I associated myelopathy. AB - We report the immunological studies of 50 cases with a chronic progressive myelopathy associated with elevated antibodies to human T-cell lymphotropic virus type I (HTLV-I) and adult T-cell leukemia-like cells (HAM). These are as follows: (1) T-cell activation; (2) increase of the OKT4/OKT8 ratio, due mostly to increased inducer/helper T cells and/or decreased suppressor/cytotoxic T cells; (3) decrease of natural killer cell numbers and activity; and (4) increase of immunoglobulins (IgG, IgA). Our results provide evidence that autoimmune events may participate in the pathogenesis of HAM and may be relevant in the hypothesis of common immune mechanisms between HAM and HTLV-I positive tropical spastic paraparesis. PMID- 2979612 TI - Identification of an erythrocyte autoantigen using monoclonal autoantibodies induced by immunization of mice with rat erythrocyte. AB - An erythrocyte autoantigen has been identified by means of monoclonal autoantibodies raised by immunizing mice with rat red blood cells (RBC). The autoantibodies reacted with intact rat and mouse RBC as judged by a cellular radioimmunoassay, and with a 52K band on western blots of rat and mouse RBC. They did not react with intact sheep RBC or blotted sheep erythrocyte membranes. Although anti-rat erythrocyte antibodies did react with bands in the molecular weight region of 50-55 K (on blots of rat erythrocyte membranes), these bands were susceptible to neuraminidase digestion, thus distinguishing them from the 52 K band recognized by monoclonal autoantibodies. The implications of the above results for the known autoantibody specificity of suppression is discussed, and it is suggested that they favour the existence of autoantigen-specific suppressor cells. PMID- 2979613 TI - Ageing and autoantibodies. AB - Immune responsiveness decreases with age, while the production of autoantibodies increases, indicating that these are end results of perturbations in the regulatory mechanisms of the immune system. The effects of ageing on the immune system are reviewed. PMID- 2979614 TI - T-cell induction of autoimmunity. PMID- 2979615 TI - Experimental autoimmune glomerulonephritis in chickens: I. Influence of source of antigen, dose and adjuvant. AB - Autoimmune glomerulonephritis was induced in chickens by immunization with human, bovine, turkey or chicken glomeruli in CFA. The influence of dose and of the pertussis vaccine was evaluated in two other separate experiments. The highest incidence of glomerulonephritis was observed in chickens immunized with human glomeruli, followed in descending order by bovine, chicken and turkey glomeruli. Capsular adhesions were seen in the last three groups. Epithelial crescents were seen only in the group immunized with chicken glomeruli. Interstitial lymphocytic infiltration was seen in 54% and 71% of chickens immunized with bovine and chicken glomeruli, respectively. The severity of the disease increased with time and the number of immunizations. IgG deposits on the GBM were seen in 92% (22/24), 54% (13/24), 50% (12/24) and 33% (8/24) of animals immunized with human, bovine, turkey and chicken glomeruli, respectively. Severity of disease was directly related to the amount of glomeruli injected. However, IgG deposits showed an inverse correlation with antigen dose. The disease was always less severe in chickens immunized with bovine glomeruli-CFA plus pertussis, as compared to bovine glomeruli-CFA immunized chickens. These studies demonstrate that the type of GBM antigen and immunization method play a significant role in the resultant histologic lesions in this model, and define conditions for optimal production of disease. PMID- 2979616 TI - Local autoimmune reaction and behavioral abnormalities after repeated nerve injury: an experimental study. AB - Sciatic nerves of rats were submitted to single and repeated injuries. The animals' gait was used to study motor function; autophagia was used to study sensory effects. An association with local post-traumatic autoimmune reaction was sought, after histochemical studies of the nerves. Our results indicated that a preliminary nerve lesion had a slight but significant influence on the recovery observed after a subsequent injury: after secondary damage, motor recovery was slightly delayed and sensory abnormalities were suggested by significant increase in autophagia. A causal role of autoimmunization was suggested but not proven. An interpretation of autophagia is proposed. PMID- 2979617 TI - Human monoclonal antibodies to phenolic glycolipid-1 from leprosy patients cross react with poly(ADP-ribose), polynucleotides and tissue bound antigens. AB - Antibodies which bind to poly(ADP-ribose) have been described in Systemic Lupus Erythematosus (SLE) and a variety of infectious diseases. Two IgM kappa human monoclonal antibodies (MAbs), TH3 and PR4, produced from the fusion of peripheral blood lymphocytes of leprosy patients with the GM4672 lymphoblastoid cell line, were found to bind to poly(ADP-ribose) in direct binding and inhibition ELISAs. Significant inhibition of binding of these MAbs to poly(ADP-ribose) occurred with phenolic glycolipid-1, the M. leprae specific glycolipid, ssDNA, dsDNA, poly(dT), as well as poly(ADP-ribose) itself. Up to 80% of binding of TH3, and 90% of binding of PR4, to poly(ADP-ribose) was inhibited by 10 mcg of ssDNA suggesting that there may be sharing of some conformational determinants. Although the serological binding profiles of TH3 and PR4 are similar, only PR4 was found to bind to basal keratinocytes of normal human interfollicular epidermis and astrocyte cytoplasm in normal brain tissue. These results support the concept that an antibody binding site may accommodate more than one epitope. Furthermore, small differences in antigen binding potential may distinguish relatively innocuous antibodies from those which may be more pathogenic. PMID- 2979618 TI - Inherited susceptibility to insulin-dependent diabetes is associated with HLA DR1, while DR5 is protective. AB - Of the HLA allelic associations with insulin-dependent diabetes (IDD) reported to date. DR3 and DR4 have been the most positive and DR2 the most negative. In 952 Caucasian proband patients reported here, only 57 or 6% had no DR3 or DR4 alleles. When these 57 patients were compared to 249 Caucasian controls similarly lacking DR3 and DR4 antigens, there were excesses of DR1 (P = 0.13) and DRW8 (P = 0.01) and deficiencies of DR2 (P = 0.03) and DR5 (P = 0.03) in the patient group. The most common phenotype in this group of patients was DR1/DR7 (12.3%). Only four DR-homozygous patients involving alleles other than DR3 and DR4 were found by genotyping, and all were DR1 homozygotes. Among 506 patients wuth DR3/DRX or DR4/DRX phenotypes, DR1 was more frequent (P = 0.001; Bonferronni P = 0.006), and DR2 (P = 0.001) and DR5 (P = 0.001) less frequent than 243 HLA-matched controls. Of 187 patients with a single DR3 and no DR4, DR1 was more frequent (P = 0.02), with DR2 (P = 0.001) and DR5 (P = 0.02) less frequent than 94 HLA DR-compatible controls. Among 319 patients with a single DR4 but no DR3, DR1 was again more frequent (P = 0.01) and DR2 (P = 0.001) and DR5 (P = 0.001) less frequent than 149 HLA-matched controls. We conclude that DR1 is an additional risk DR allele for IDD to that of DR3 and DR4, and DR5 an additional protective DR allele to that of DR2. PMID- 2979619 TI - Autoantibody induced by experimental ocular Onchocerca infection: identification and characterization of autoantigens. AB - Hartley guinea pigs were injected with Onchocerca lienalis microfilariae (Mf) as a model for human infection with Onchocerca volvulus. Subconjunctival injection of guinea pigs with O. lienalis Mf results in penetration of Mf to the central cornea, and formation of acute inflammatory reactions resembling those of human onchocerciasis around dead Mf. Sera from these animals contain IgG autoantibodies directed against at least two components of 3M KC1 extracts of guinea pig cornea and liver. Components with apparent molecular weights of 94-99 and 110-120 kilodaltons (KD) were recognized in both cornea and liver extracts, while 100, 74 and 34 kD components were recognized in liver extracts only. Components of 140 kD and 84 kD were recognized in cornea extracts only. The guinea pig liver extract cross-reacted with homogenate antigens from O. lienalis Mf, while the guinea pig cornea extract did not. Autoantibody-mediated inflammatory mechanisms may contribute to immunopathologic processes in onchocerciasis. PMID- 2979620 TI - Increase of class II HLA molecules on the membrane of B lymphocytes from patients with rheumatoid arthritis. AB - Using a novel cytofluorometric method of cellular antigen quantification, we examined peripheral blood mononuclear cells (PBMC) from patients suffering from rheumatoid arthritis (RA) for quantitative modification of class II human leucocyte antigen (HLA) molecules expressed on the surface. Class II HLA molecules were detected by indirect immunofluorescence with a monomorphic monoclonal antibody. No change was observed in the density of class II HLA molecules at the surface of monocytes of RA patients as compared to that of paired healthy subjects. We confirmed that the percentage of class II HLA-bearing T cells was slightly increased in RA patients versus controls, but the density of class II antigens per cell could not be determined accurately. An increase in the density of class II HLA molecules on RA B cells was shown, suggesting that a chronic activation stage of this population contributes to the disease. PMID- 2979621 TI - Complement-fixing gastric parietal cell autoantibodies. A good marker for the identification of type A chronic atrophic gastritis. AB - Using an indirect immunofluorescence (IIF) technique, gastric parietal cell autoantibodies of IgG class (GPCA-IgG) were found in 2% of a normal population, in 5-26% of organ-specific autoimmune subjects and in 100% of patients with pernicious anaemia. With the exception of subjects with alopecia, there was a significantly increased prevalence of GPCA-IgG in autoimmune patients with respect to normal controls. GPCA of IgA class were detected in 22% of GPCA-IgG positive subjects, whereas GPCA of IgM class were uncommon. One-hundred and fifteen subjects underwent gastroscopy and body mucosal biopsy. Histopathological findings of chronic atrophic gastritis (CAG) were present in 71% of GPCA-IgG positive autoimmune patients without pernicious anaemia, in 100% of GPCA-IgG positive patients with pernicious anaemia, and in 20% of GPCA negative autoimmune patients. Complement-fixation test was performed in 46 GPCA-IgG positive subjects without pernicious anaemia using the IIF method. Twenty-nine patients (63%) were found to fix complement fractions till C9 (CF-GPCA) together with properdin factor, and in 25 of them (86%) the histological examination of body gastric mucosa disclosed a CAG (P = 0.0003 versus GPCA-IgG positive/CF negative controls). No significant difference was observed for the prevalence of CAG in GPCA-IgG positive/CF negative subjects with respect to GPCA-IgG negative control group. We conclude that the presence of CF-GPCA represents a useful immunological marker in the identification of CAG, while no predictive value seems to be associated with non-complement fixing GPCA-IgG. PMID- 2979622 TI - On the relationship between islet cell antibodies and insulin autoantibodies in patients at risk from insulin dependent diabetes. AB - The frequencies of islet cell antibodies (ICA) and insulin autoantibodies (IAA) were studied in three clinically well defined groups, using an aprotinin sensitised indirect immunofluorescence assay for ICA and a direct binding solid ELISA for IAA, and the association of these two serological markers for insulin dependent diabetes analysed. Frequency of ICA was 10.7% in siblings of diabetics, 15.5% in discordant identical twins and 65.9% in newly diagnosed diabetic patients. Frequency of IAA was 7.1% in siblings, 46.7% in discordant twins and 38.6% in newly diagnosed diabetic patients. No correlation was demonstrated between the two autoantibodies in the siblings. In the newly diagnosed diabetic patients there were sera positive or negative for both, but 22 (50%) of the sera showed dissociation between the two antibodies. The studies of twins showed that IAA and ICA fluctuated independently with time, and demonstrate the inappropriateness of seeking such an association in cross-sectional surveys. An association could not be demonstrated in this group even if data from multiple samples taken at different points in time were pooled, scoring an individual as positive if at any time their sera had been positive for the corresponding antibody. Thus our data showed no correlation between ICA and IAA in any of the groups studied. PMID- 2979624 TI - Major histocompatibility complex (MHC) factors predisposing to and protecting against Graves' eye disease. AB - We investigated the influence of gene mapping within the major histocompatibility complex on the susceptibility to Graves' eye disease. We studied 133 randomly selected patients with Graves' disease, many of whom had eye disease. HLA B8 and DR3 carried the greatest risk for disease but the difference between the two patient groups was not statistically significant. An earlier finding that Hungarian patients with a subset of B8, (B8 + DR7 +) had a greatly enhanced risk for eye disease was confirmed in Newfoundland patients. HLA B8 and DR7 are probably carried on different homologous chromosomes and their interaction enhances eye disease. HLA-DR4 was negatively correlated with eye disease. In particular, a subset of DR4 (B35 + DR4 +) appears to protect against eye disease. We have also derived the haplotypes of 22 probands, half of whom had eye disease. The haplotype data emphasized the high frequency of HLA A1 B8 DR3 C4A*QO and C4B*1 in both patient groups, 15% of the haplotypes in the group with eye disease and 25% in that without eye disease. Forty-one percent of haplotypes in the eye disease group and 32% in the no eye disease group were either C4A*QO or C4B*QO. In one proband with eye disease B8 and DR7 were carried on separate chromosomes. The phenotype DR4, C4A*3 C4B*1 was found in 3/20 haplotypes of patients without eye disease but in 0/20 of patients with eye disease. This finding is in keeping with the increased frequency of the DR4 C4A*3 C4B*1 in the patient group with no eye disease when 94 patients were phenotyped.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979623 TI - Presence of insulin autoantibodies at clinical diagnosis of diabetes mellitus type I predicts loss of beta cell function. AB - Recently the spontaneous development of insulin autoantibodies (IAA) has been detected in patients at diagnosis of Type I diabetes mellitus before the beginning of insulin treatment. The present study was undertaken to investigate if the presence of IAA at clinical onset of IDDM may act as a new marker of the beta cell function. The results obtained showed that IAA were present in 44% of newly diagnosed diabetic patients before therapy. Patients without IAA displayed a higher C-peptide secretion than those with IAA, at six months (12.11 +/- 5.08 versus 5.88 +/- 3.25 ng/ml/10 min.)(X +/- SD) and at twelve months (10.45 +/- 3.05 versus 4.90 +/- 5.25 ng/ml/10 min)(X +/- SD) of the follow up period. HbA1 levels, and insulin requirements were similar in both groups (IAA+ and IAA-). We conclude that the presence of insulin autoantibodies at clinical diagnosis, before initiating insulin treatment, may well predict the loss of the beta cell function. PMID- 2979625 TI - Meeting report: methods of detection and clinical relevance of anti DNA/ENA antibodies (Middlesex Hospital Medical School February 26th 1988). PMID- 2979627 TI - Polyclonal nature of islet cell antibodies in insulin-dependent diabetes mellitus. PMID- 2979626 TI - Pyrithioxine and hypoglycemic autoimmune syndrome. PMID- 2979628 TI - Suprachiasmatic nuclear lesions eliminate circadian rhythms of drinking and activity, but not of body temperature, in male rats. AB - Male Long-Evans rats were maintained in light proof cabinets while drinking, activity, and telemetered body temperature (Tb) data were collected. After suprachiasmatic nuclear (SCN) lesions, the rats were exposed to a 12:12 light dark cycle, a 6-hr delay in the lighting cycle, and constant dark. Lesions that abolished the drinking and activity rhythms did not eliminate the Tb rhythm. However, the amplitude, phase, and free-running period of the Tb rhythm were altered. Lesions that only partially damaged the SCN had similar, though lesser effects. In some cases, Tb rhythms remained normal, activity rhythms were only temporarily disrupted, and drinking rhythms were eliminated in the same animals. These results support the conclusion that Tb can remain rhythmic after lesions that permanently or temporarily disrupt other circadian rhythms. Of the three rhythms, it appears that drinking rhythms are most easily and Tb rhythms least easily disrupted by SCN lesions. PMID- 2979629 TI - Effects upon circadian rhythmicity of an alteration to the sleep-wake cycle: problems of assessment resulting from measurement in the presence of sleep and analysis in terms of a single shifted component. AB - Experiments were performed upon groups of three or four human subjects in an isolation chamber (total n = 14). Subjects lived initially on a conventional lifestyle and then delayed their hours of sleep by 8 hr (so mimicking some aspects of nightwork) for 2 or 5 days. They also performed two constant routines- protocols designed to minimize any effects due to the environment, mealtimes, and activity. Regular samples of urine were taken when subjects were awake, and were analyzed for sodium, potassium, and chloride; rectal temperature was measured and logged at 6-min intervals throughout. Shifts in circadian rhythms produced by the change in sleep time were assessed by cosinor and cross-correlation techniques. The protocol enabled these assessments to be made on days when sleep was allowed and under constant-routine conditions, so that masking and behavioral effects could be investigated also. The results confirmed that adjustment to the change in sleep time was slow and only partial, and that assessments made on days when sleep was allowed overestimated this adjustment. Furthermore, it was concluded that, whereas cosinor and cross-correlation techniques using only one shifting component were equally useful in describing the observed changes, both were inferior to a cross-correlation technique that made use of two shifting components. Some practical and theoretical implications of these results are discussed. PMID- 2979630 TI - Precise annual changes in the numbers of "synaptic" ribbons and spherules in the rat pineal gland. AB - Although pineal "synaptic" ribbons (SR) are frequently examined by means of quantitative electron microscopy, their functional significance remains unclear. The same is true for related structures--"synaptic" spherules (SPH). In the course of such studies, it has been noted that SR counts may differ from laboratory to laboratory. Because seasonal changes may play a role, a 2-year study was performed on male rats kept under routine laboratory conditions and killed at monthly intervals during daytime or nighttime. Both structures examined showed distinct day-night differences throughout the year, with higher numbers at night than during the day. There were significant annual changes in both SR and SPH during both daytime and nighttime. The comparison of the curves from the 2 years showed that they were virtually identical both during daytime and nighttime. The numbers of SR were the highest in October and the lowest in April; the numbers of SPH had two plateaus, one with lower values from November to April, and the other with higher values from May to October. It appears from the present study that SR and SPH numbers in the rat pineal gland show statistically significant and precisely timed seasonal changes that may well account for the variations of SR numbers in the different publications. PMID- 2979631 TI - Cell cycle synchronization of Gonyaulax polyedra by filtration: quantized generation times. AB - A size filtration method to synchronize cultures of the dinoflagellate Gonyaulax polyedra to the beginning of the G1 phase has been developed. This technique selects newly born cells by two sequential filtrations, based on the fact that cell division is restricted to the beginning of the day, so that a decrease in cell volume occurs at this time. The fraction of synchronized cells immediately after the second filtration is about 90%; the procedures do not alter the free running period or phase of glow rhythm, and the selected cells divide again in a few days. Applying this method, we have found that the generation times of this species in a light-dark cycle (LD 12:12) are indeed quantized to multiples of 24 hr, but are variable from generation to generation. PMID- 2979632 TI - Axon-sparing lesions of the hypothalamic paraventricular nucleus abolish gonadal responses to photoperiod in male Syrian hamsters. AB - Intrahypothalamic injections of the excitotoxin N-methyl-D,L-aspartic acid (NMA) that destroyed the parvocellular regions of the paraventricular nucleus (PVN) prevented gonadal regression in male hamsters (Mesocricetus auratus) kept in short days (LD 6:18). After 17 weeks in short days, these NMA-injected animals had large, functional gonads and displayed all components of male sexual behavior. These findings confirm that neurons of the PVN are part of the pathway that mediates photoperiodism in hamsters. PMID- 2979633 TI - Comparative anatomy of the mammalian hypothalamic suprachiasmatic nucleus. AB - A detailed analysis of the cytoarchitecture, retinohypothalamic tract (RHT) projections, and immunohistochemical localization of major cell and fiber types within the hypothalamic suprachiasmatic nuclei (SCN) was conducted in five mammalian species: two species of opossum, the domestic cat, the guinea pig, and the house mouse. Cytoarchitectural and immunohistochemical studies were conducted in three additional species of marsupial mammals and in the domestic pig. The SCN in this diverse transect of mammalian taxonomy bear striking similarities. First, the SCN are similar in location, lying close to the third ventricle (3V) dorsal to the optic chiasm (OC), with a cytoarchitecture characterized by small, tightly packed neurons. Second, in all groups studied, the SCN receive bilateral retinal input. Third, the SCN contain immunohistochemically similar elements. These similarities suggest that the SCN developed characteristic features early in mammalian phylogeny. Some details of SCN organization vary among the species studied. In marsupials, vasopressin-like immunoreactive (VP-LI) and vasoactive intestinal polypeptide-like immunoreactive (VIP-LI) cells codistribute primarily in the dorsomedial aspects of the SCN, while in eutherians, VP-LI and VIP-LI cells are separated into SCN subnuclei. Furthermore, the marsupial RHT projects to the periventricular dorsomedial region, whereas the eutherian RHT projects more ventrally in the SCN into the zone that typically contains VIP-LI perikarya. PMID- 2979634 TI - Phototherapy for seasonal affective disorder. PMID- 2979635 TI - Winter depression and the phase-shift hypothesis for bright light's therapeutic effects: history, theory, and experimental evidence. PMID- 2979636 TI - Neurobiology of seasonal affective disorder and phototherapy. PMID- 2979637 TI - On the question of mechanism in phototherapy for seasonal affective disorder: considerations of clinical efficacy and epidemiology. PMID- 2979638 TI - The photoperiodic phenomena: seasonal modulation of the "day within". PMID- 2979639 TI - Seasonal affective disorder, hibernation, and annual cycles in animals: chipmunks in the sky. PMID- 2979640 TI - Seasonal affective disorders: animal models non fingo. PMID- 2979641 TI - Chronic clorgyline treatment of Syrian hamsters: an analysis of effects on the circadian pacemaker. AB - Clorgyline, a type A monoamine oxidase inhibitor with antidepressant properties when administered to depressed patients, is often associated with disturbances of the human sleep-wake cycle. In order to assess its effects on the mammalian circadian system, this drug was administered chronically to Syrian hamsters. It was found to affect the hamster circadian system in four specific ways. Clorgyline increased the intrinsic period of wheel-running activity, altered the phase response curve to brief light pulses, altered the reduced waveform of running activity in animals maintained in light-dark cycles or constant darkness, and increased the activity-rest ratio in animals maintained in constant darkness. Our data support the interpretation that clorgyline exhibits direct or indirect input to the circadian pacemaker and alters the processing of photic information to the pacemaker. PMID- 2979642 TI - Does a biological clock reside in the eye of quail? AB - The site (intra- vs. extraocular) of the circadian clock driving an ocular melatonin rhythm in Japanese quail was investigated by alternately covering the left and right eyes of individual quail, otherwise held in constant light (LL), for 12-hr periods. This procedure exposed each eye to a light-dark (LD) 12:12 light cycle 180 degrees (12 hr) out of phase with the LD 12:12 light cycle experienced by the other eye. This protocol entrained the melatonin rhythm in the left eye of quail 180 degrees out of phase with the rhythm expressed in the right eye. These results are compatible with the hypothesis that an independent light entrainable circadian pacemaker resides in each eye; they are incompatible with the hypothesis that a single (or functionally single) extraocular pacemaker drives the ocular melatonin rhythm in both eyes. However, the results are also compatible with a model in which two independent extraocular circadian pacemakers, each with an exclusive photic input from one eye, drive the ocular melatonin rhythm. PMID- 2979643 TI - The effects of intraventricular carbachol injections on the free-running activity rhythm of the hamster. AB - The effects of light on the circadian pacemaker in the suprachiasmatic nucleus (SCN) are mediated by the retinohypothalamic tract (RHT) and by the retinogeniculosuprachiasmatic tract (RGST). The neurotransmitter of the RGST is neuropeptide Y. The RHT may contain glutamate and aspartate. Recent evidence indicates that acetylcholine could also be involved in phase shifting by light. We determined that intraventricular injections with an acetylcholine agonist, carbachol, induces phase advances during the subjective day and phase delays during the early subjective night. No differences were observed between phase shifts induced in constant darkness and those induced in continuous light. A dose response curve for carbachol was described at circadian time 6 (CT6). Injections at CT14 with various dosages of carbachol indicated the same dose dependency for this circadian time. Finally, carbachol injections in split animals resulted in similar responses of the two components of the split activity rhythm. PMID- 2979644 TI - Pineal and photoperiodic influences on fat deposition, pelage, and testicular activity in male meadow voles. AB - Pinealectomy completely prevented gonadal regression as well as reduction in body weight and white adipose tissue content of the gonadal and retroperitoneal fat deposits in male meadow voles transferred from long to short day lengths. Pineal influences on pelage characteristics depended on which parameter was assessed. For instance, the increase in guard hair length observed in short-day control voles was blocked by pinealectomy; however, a similar increase in underhair length was unaffected by removal of the pineal gland. Photoperiod-dependent changes in fat deposition, testicular activity, and guard hair length presumably rely on altered pineal secretory activity to transduce the effects of day length on the neuroendocrine axis; however, mechanisms independent of pineal activity may be capable of mediating photoperiodic control of underhair growth. PMID- 2979645 TI - Different responses of the circadian system to GABA-active drugs in two strains of mice. AB - Recent work in our laboratory has shown that sodium pentobarbital injections can induce phase-dependent phase shifts of the circadian rhythm of locomotor activity with the maximum advance at circadian time (CT) 8 and the maximum delay at CT0 in SK/Nga mice but no phase shifts in C57BL/6 mice. In the present study, the possibility that the differences in the effects of pentobarbital on the circadian rhythm may be due to different contributions of the GABA-ergic system to circadian organization in the two strains was tested by comparing the responses of SK mice with those of C57BL mice to muscimol (2 mg/kg), a GABA receptor agonist, and triazolam (25 mg/kg), which is thought to act by potentiating the action of GABA. The hypothesis that pentobarbital-induced phase shifts of SK mice are mediated by the GABA receptor system was also tested by observing whether the phase-shifting effects of pentobarbital were blocked by bicuculline (0.5 mg/kg), a selective antagonist of GABA, injected 3 min prior to pentobarbital (30 mg/kg). The results indicated that muscimol induced phase advances at CT8 and phase delays at CT0, and triazolam induced phase advances at CT8 in SK mice. No phase shifts were induced by any treatment in C57BL mice. These results suggest that the role of GABA-ergic systems in circadian organization may be different in SK and C57BL mice. In addition, bicuculline could block the phase-shifting effects of pentobarbital in SK mice, suggesting that the GABA receptor system may mediate phase-shifting effects of pentobarbital in SK mice. PMID- 2979646 TI - Effects of constant darkness and constant light on circadian organization and reproductive responses in the ram. AB - The relationship between circadian rhythms in the blood plasma concentrations of melatonin and rhythms in locomotor activity was studied in adult male sheep (Soay rams) exposed to 16-week periods of short days (8 hr of light and 16 hr of darkness; LD 8:16) or long days (LD 16:8) followed by 16-week periods of constant darkness (dim red light; DD) or constant light (LL). Under both LD 8:16 and LD 16:8, there was a clearly defined 24-hr rhythm in plasma concentrations of melatonin, with high levels throughout the dark phase. Periodogram analysis revealed a 24-hr rhythm in locomotor activity under LD 8:16 and LD 16:8. The main bouts of activity occurred during the light phase. A change from LD 8:16 to LD 16:8 resulted in a decrease in the duration of elevated melatonin secretion (melatonin peak) and an increase in the duration of activity corresponding to the changes in the ratio of light to darkness. In all rams, a significant circadian rhythm of activity persisted over the first 2 weeks following transfer from an entraining photoperiod to DD, with a mean period of 23.77 hr. However, the activity rhythms subsequently became disorganized, as did the 24-hr melatonin rhythms. The introduction of a 1-hr light pulse every 24 hr (LD 1:23) for 2 weeks after 8 weeks under DD reinduced a rhythm in both melatonin secretion and activity: the end of the 1-hr light period acted as the dusk signal, producing a normal temporal association of the two rhythms. Under LL, the 24-hr melatonin rhythms were disrupted, though several rams still showed periods of elevated melatonin secretion. Significant activity rhythms were either absent or a weak component occurred with a period of 24 hr. The introduction of a 1-hr dark period every 24 hr for 2 weeks after 8 weeks under LL (LD 23:1) failed to induce or entrain rhythms in either of the parameters. The occurrence of 24-hr activity rhythm in some rams under LL may indicate nonphotoperiodic entrainment signals in our experimental facility. Reproductive responses to the changes in photoperiod were also monitored. After pretreatment with LD 8:16, the rams were sexually active; exposure to LD 16:8, DD, or LL resulted in a decline in all measures of reproductive function. The decline was slower under DD than LD 16:8 or LL.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2979647 TI - Circadian rhythm resetting in sparrows: early response to doublet light pulses. AB - Circadian responses were studied using the perching activity of house sparrows (Passer domesticus). The sparrows were subjected to single or double 4-hr light pulses (the single pulses or the second pulses of the doublets scanned 24 hr) in the first cycle after previous entrainment to a light-dark cycle (LD 12:12). The differences in times at which the birds commenced perch-hopping in LD 12:12 before the pulses and in the five cycles immediately following the pulses were determined (phase shifts). A 24-hr time profile for phase shifts in response to single light pulses replicated our previous study: Early-night pulses delayed the rhythm (-1.7 hr), while late-night pulses advanced the rhythm (+3.8 hr). After pretreatment with a light pulse that advanced the birds +2.7 hr, the resetting curve was advanced. There were no delays; the range of average shifts was +0.1 hr to +6.2 hr. After pretreatment with a light pulse that delayed the birds -1.7 hr, the resetting curve was delayed. Average delays as much as -1.1 hr and advances up to +2.1 hr were measured. The data for double pulses were interpreted from predictions made from single-pulse data. PMID- 2979648 TI - Diapause induction and termination: north-south strain differences in Ostrinia nubilalis. AB - Diapause induction and termination responses of a northern strain (Minnesota [MN]) of Ostrinia nubilalis were compared with those of a southern strain (Georgia [GA]). A thermoperiod in constant light (12 hr at 25 degrees C alternating with 12 hr at 4 degrees C) failed to induce diapause in GA larvae, but approximately 50% diapause induction was observed in the MN population. Moreover, the 50% of MN larvae that continued their development (i.e., underwent pupation and adult development) did so at a slower rate, as measured by days to pupation, than GA larvae. In the laboratory, diapausing MN larvae responded more slowly to the optimal light-dark (LD) cycle for terminating diapause, LD 16:8, than did GA larvae. In the field MN populations are univoltine (i.e., are characterized by one generation per year). A delayed termination response in the spring, coupled with a longer critical daylength for diapause induction as daylength decreases during late summer (earlier diapause) restricts the time during which development can occur as contrasted with GA populations. In addition, it is postulated that these two phenomena, coupled with a possibly slower growth rate in the MN insects as revealed under laboratory conditions, may collectively represent the basis for univoltinism in the field. PMID- 2979649 TI - Serotonin phase-shifts the circadian rhythm of locomotor activity in the cockroach. AB - Serotonin, a putative neurotransmitter in insects, was found to cause consistent phase shifts of the circadian rhythm of locomotor activity of the cockroach Leucophaea maderae when administered during the early subjective night as a series of 4-microliters pulses (one every 15 min) for either 3 or 6 hr. Six-hour treatments with dopamine also caused significant phase shifts during the early subjective night, but 3-hr treatments with dopamine had no phase-shifting effect. Other substances tested in early subjective night (norepinephrine, octopamine, gamma-aminobutyric acid, glutamate, carbachol, histamine, tryptophan, tryptamine, N-acetyl serotonin, or 5-hydroxyindole-3-acetic acid) did not consistently cause phase shifts. The phase-shifting effect of serotonin was found to be phase dependent. The phase response curve (PRC) for serotonin treatments was different from the PRC for light. Like light, serotonin caused phase delays in the late subjective day and early subjective night, but serotonin did not phase-shift rhythms when tested at phases where light causes phase advances. PMID- 2979650 TI - The intergeniculate leaflet partially mediates effects of light on circadian rhythms. AB - Photic signals affect circadian activity rhythms by both phasic and tonic mechanisms that modulate pacemaker phase and period. In mammals, the effects of light on circadian activity are mediated by the retina, which communicates with the suprahiasmatic nucleus (SCN) by two different anatomical routes: the retino hypothalamic tract (RHT), originating in the retina, and the geniculo hypothalamic tract (GHT), arising from a retino-recipient nucleus, the intergeniculate leaflet (IGL). We assessed the roles of these two afferent systems in mediating phasic and tonic effects of light on circadian activity in IGL-lesioned animals. Destruction of the IGL significantly affected phase shifts produced by brief light pulses (phasic effect) and modified the change in period (tau) of the free-running activity rhythm produced by changing the level of constant light (LL) (tonic effect). Phase advances produced by brief light pulses were decreased in amplitude while phase delays were increased in IGL-lesioned animals as compared to controls. The free-running period in constant dark (tau DD) of IGL-lesioned animals was greater than tau DD of controls, and the lengthening of tau normally produced by LL was not observed or was greatly reduced in IGL-lesioned animals. Entrainment to light-dark cycles was unaffected by the lesions, as were other aspects of the circadian activity rhythm that normally change in response to LL (e.g., activity-rest ratio, total activity, splitting). Our data support the interpretation that the IGL plays a significant role in relaying information regarding illumination intensity to the SCN. PMID- 2979651 TI - Circadian rhythmicity within single cells of Paramecium bursaria. AB - Cell populations of Paramecium bursaria show mating reactivity in the light period, but not in the dark period, when exposed to a light-dark cycle (LD 12:12). After they are transferred to constant-light (LL) conditions (1,000 lux), they continue to show a circadian rhythm of mating reactivity. The rhythm gradually dampens in LL so that mating reactivity in populations becomes arrhythmic in LL within 2 weeks. We wanted to know whether the arrhythmicity of this population was due to the absence of circadian rhythmicity within each individual cell, or merely due to asynchrony of a population of individually rhythmic cells. Therefore, single cells were isolated randomly from an arrhythmic population that had been in LL for a long time. Then the mating reactivity of these single cells was individually tested every 3 hr for 2 days. Each single cell showed a circadian mating rhythm in LL. This shows that the disappearance of the mating rhythm in cell populations under LL is not caused by disappearance of circadian rhythmicity within individual cells, but is due to desynchronization among cells in a population. When an arrhythmic population in LL is darkened for 9 hr, the mating reactivity rhythm of the cell population reappears. This occurs by resynchronization of the rhythms among individual cells, as can be shown by exposing single cells to pulses of 9 hr of darkness. This dark treatment causes phase shifts of single-cell rhythms, and a phase response curve is obtained for this stimulus. This phase-shifting behavior explains the efficacy of 9-hr dark pulses in restoring the population's rhythm. PMID- 2979652 TI - Further evidence that the circadian clock in Drosophila is a population of coupled ultradian oscillators. AB - We hypothesize that ultradian oscillators are coupled to yield a composite circadian clock in Drosophila. In such a system, period would be a function of the tightness of coupling of these oscillators, increasing as coupling loosens. Ultradian oscillations would become apparent under weak coupling or in the absence of coupling. A new technique for calculating signal-to-noise ratio (SNR) for biological rhythms to characterize their precision has yielded support for this hypothesis. SNR of rhythms of the allelic series of mutations at the period (per) locus of Drosophila melanogaster were compared. Per(o) was the noisiest, grading through perL, per+, and pers, the least noisy. SNR decreases significantly with increasing period in pers, per+, and perL; per(o) typically has multiple ultradian oscillations and the lowest SNR. At least 70% of perL individuals also exhibit ultradian periodicities. PMID- 2979653 TI - Differential effects of pinealectomy on circadian rhythms of feeding and perch hopping in the European starling. AB - To study the effects of pinealectomy on the circadian rhythms of locomotor activity and feeding. European starlings (Sturnus vulgaris) were held in constant light (0.2 lux and 200 lux) and under constant temperature conditions. Locomotor activity was measured by means of perches with microswitches mounted underneath, and feeding with an infrared photocell system at the feeder. Pinealectomy consistently led to disturbances in perch-hopping rhythms and often to a complete loss of rhythmicity as revealed by periodogram analysis. In some birds, perch hopping rhythms recovered following a period of initial arrhythmicity. When a perch-hopping rhythm was present, its period was usually shorter than it had been before pinealectomy. In contrast to its effects on perch hopping, pinealectomy had no effect on the persistence of feeding rhythmicity, although its period, like that of the hopping rhythm, decreased after this operation. These results support the hypothesis derived from previous studies that the circadian organization of feeding is different from that of perch hopping. Different circadian pacemakers may be involved, but other models may possibly explain the data just as well. PMID- 2979654 TI - Are protein synthesis inhibition and phase shifting of the circadian clock in Gonyaulax correlated? AB - We describe a method whereby the effect of protein synthesis inhibitors upon protein synthesis in Gonyaulax cultures may be reliably measured. Using this method, we found that protein synthesis inhibition and clock resetting were correlated, but that the correlation was not as close as has been reported in other systems. The effect of the inhibitors anisomycin and cycloheximide upon phase shifting of the circadian clock was a function of the illumination and temperature conditions to which the cells were subjected, but these factors did not appear to influence the inhibition of protein synthesis by these drugs. Cellular protein synthesis did not recover immediately from the inhibitors' effects; depending upon the previous concentration of the inhibitor, translational recovery from the drugs may require hours. This observation has important implications for the analysis of any phase response curve when the stimulus is a chemical. PMID- 2979655 TI - Rapid photoperiodic responses in Japanese quail: is daylength measurement based upon a circadian system? AB - Experimental photoperiods, presented either once only or repeatedly, were used to assess the oscillatory and hourglass properties of the photoperiodic clock in Japanese quail. Gonadectomized quail on 8-hr daylengths respond to a single skeleton photoperiod consisting of two 8-hr light pulses separated by 2 hr of darkness (i.e., LDLD 8:2:8:6) with a marked increase in secretion rate of luteinizing hormone (LH). This response suggests that the second light pulse interacts with a "photoinducible phase" (phi i) lying some 10-16 hr from "dawn" (start of the first light pulse). If, however, groups of quail maintained on 8-hr daylengths are transferred to continuous darkness (DD), and the position of the phi i is sought by a single 8-hr light pulse applied at various times on the first or third day of DD, then an increase in circulating LH is, at best, barely detectable. It would appear that a strongly responsive phi i does not recur rhythmically in DD. Instead, the light pulse apparently acts primarily as a "dawn" signal that triggers a single cycle of photoinducibility, since a second 8 hr light pulse, placed to begin 2 hr after the end of the first, induces a large increase in plasma LH. Similar results are obtained if any single 8-hr light pulse presented to animals held in darkness is preceded, 10 hr earlier, by a short "dawn" light signal. Such dawn signals can be effective when very short; a pulse of only 30 sec can cause a subsequent phi i. The dawn pulse is effective at any circadian phase and leads to a single cycle in photoinducibility. In contrast, a much longer light pulse (perhaps not less than 4 hr) is needed to interact with phi i if significant gonadotropin secretion is to be stimulated. In confirmation of the findings described above, we found that Nanda-Hammer lighting schedules have remarkably little effect in stimulating gonadotropin secretion in gonadectomized quail. There is, for example, a very marked difference between the effectiveness of "resonating" schedules such as LD 6:6, which stimulates a high LH secretion rate since each "inductive" light pulse is preceded by an appropriate "dawn" signal, and a theoretically effective schedule such as LD 6:30, which induces a very small response by comparison. Such schedules (even theoretically noninductive ones) can, however, be made very highly inductive if alternate light pulses are preceded by an appropriately positioned 15-min light pulse to act as "dawn." PMID- 2979656 TI - The influence of prior wakefulness on REM sleep. AB - Data from studies of naps and of shifted sleep were used to determine the relationship between two measures of rapid eye movement (REM) sleep (percentage of REM in the first 2 hr of sleep and REM latency) and prior wakefulness. For each sample, we calculated the difference between the observed value and that predicted by a cosine function that estimated the circadian rhythm of REM sleep propensity. The difference values were found to correlate reliably with hours and log hours of prior wakefulness. We conclude that while REM sleep is regulated in part by an endogenous circadian oscillator, it is also influenced by the duration of prior wakefulness. PMID- 2979657 TI - Dynamics of discrete entrainment of the circadian rhythm in the rat pineal N acetyltransferase activity during transient cycles. AB - To elucidate entrainment of a pacemaker controlling the N-acetyltransferase (NAT) rhythm in the rat pineal gland, we studied the phase response curves (PRCs) of this rhythm. We exposed 50- to 60-day-old male Wistar rats maintained in a light dark cycle (LD 12:12) to a 1-min light pulse at different times before midnight or at various times throughout the whole night. We then released them into constant darkness and studied the morning NAT decline during the night when rats were pulsed before midnight, as well as the evening NAT rise and the morning decline after 4 days following the pulses. The PRC for the first NAT decline and the PRCs for the NAT rise and decline after 4 days were compared with published transient PRCs (Illnerova and Vanecek, 1982b), in order to obtain a complete picture of the dynamics of the NAT rhythm entrainment during the transient cycles. Phase delays in the NAT rise due to a pulse before midnight were complete (i.e., identical to those of day 4) on day 1. Phase delays in the NAT decline were almost complete on day 1, while incomplete phase delays were observed on day 0. Phase advances in the NAT rise and decline due to a pulse past midnight had different dynamics: Advances in the decline were complete on day 1, while advances in the rise were absent on day 1 and much smaller than in the decline on day 4. The results are discussed in terms of a two-component (E-M) pacemaker controlling the NAT rhythm. The NAT rise may reflect the phase of the E component, while the decline reflects the M-component. Phase delays of the E component are accomplished within one cycle, and so are phase advances of the M component. However, although delays of E already result in delays of M one cycle after the pulse, it takes several transient cycles before advances of M begin to induce advances of E. PMID- 2979658 TI - Genetic and molecular analysis of biological rhythms. PMID- 2979659 TI - Pineal melatonin rhythms in the lizard Anolis carolinensis: I. Response to light and temperature cycles. AB - Both light and temperature can influence the pineal's synthesis of the indoleamine melatonin. An investigation of the effects of light and temperature cycles on the pineal melatonin rhythm (PMR) showed the following: (1) Both daily light cycles and daily temperature cycles could entrain the PMR; melatonin levels peaked during the dark phase of a light-dark cycle or the cool phase of a temperature cycle. (2) The PMR could be entrained by a temperature cycle as low as 2 degrees C in amplitude in lizards held in constant light or constant darkness. (3) The length of the photoperiod or thermoperiod affected the phase, amplitude, or duration of the PMR. (4) When presented together, the effects of light and temperature cycles on the PMR depended on the phase relationship between the light and temperature cycles, as well as on the strength of the entraining stimuli, such as the amplitude of the temperature cycle. (5) Exposure to a constant cold temperature (10 degrees C) eliminated the PMR, yet a rhythm could still be expressed under a 24-hr temperature cycle (32 degrees C/10 degrees C), and the rhythm peaked during the 10 degrees C phase of the cycle. (6) A 6-hr dark pulse presented during the day did not elicit a premature rise in melatonin levels. These studies show how environmental stimuli can control the pineal rhythm of melatonin synthesis and secretion. Previous studies have supported a model in which the lizard's pineal acts as a circadian pacemaker within a multioscillator circadian system, and have implicated melatonin as a hormone by which the pineal may communicate with the rest of the system. The lizard pineal, therefore, may act as a photo- and thermoendocrine transducer translating light and temperature information into an internal cue in the form of the PMR. The PMR, in turn, may control the phase and period of circadian clocks located elsewhere, insuring that the right internal events occur at the right time of day. PMID- 2979660 TI - Pineal melatonin rhythms in the lizard Anolis carolinensis: II. Photoreceptive inputs. AB - The pineal organ of the lizard Anolis carolinensis acts as a transducer of photoperiodic information, since light can affect the pineal melatonin rhythm (PMR). The synthesis and secretion of melatonin may be a major mechanism whereby a circadian pacemaker within the pineal can control circadian clocks located elsewhere. An investigation into potential routes by which light could affect the PMR showed that (1) removal of the photosensory parietal eye did not affect the PMR as compared to controls under either a light-dark (LD) 12:12 cycle and a constant temperature (32 degrees C) or an LD 12:12 cycle and a daily temperature cycle (32 degrees C/20 degrees C); (2) removal of both the parietal eye and the lateral eyes did not affect the PRM of anoles held in LD 12:12 (constant 32 degrees C); (3) the PMR of blinded anoles re-entrained to a 10-hr shift in the phase of the LD cycle as rapidly as that of sighted anoles; (4) blocking light penetration to the brains of anoles, but leaving the lateral eyes exposed, blocked the ability of anoles to re-entrain to a 10-hr shift in the phase of an LD cycle. The data support the hypothesis that light directly affects the PMR in Anolis and that other potential photic inputs (parietal eye, lateral eyes) play little or no role. This conclusion is supported by previous neurophysiological and ultrastructural studies showing that the lizard pineal possesses functional photoreceptors. PMID- 2979661 TI - EEG power density during nap sleep: reflection of an hourglass measuring the duration of prior wakefulness. AB - The relation between the duration of prior wakefulness and EEG power density during sleep in humans was assessed by means of a study of naps. The duration of prior wakefulness was varied from 2 to 20 hr by scheduling naps at 1000 hr, 1200 hr, 1400 hr, 1600 hr, 1800 hr, 2000 hr, and 0400 hr. In contrast to sleep latencies, which exhibited a minimum in the afternoon, EEG power densities in the delta and theta frequencies were a monotonic function of the duration of prior wakefulness. The data support the hypothesis that EEG power density during non rapid eye movement sleep is only determined by the prior history of sleep and wakefulness and is not determined by clock-like mechanisms. PMID- 2979662 TI - Circannual variations in plasma luteinizing hormone levels in castrated male European starlings (Sturnus vulgaris). AB - Intact and castrated male European starlings were held for about 2 years in a constant 12-hr photoperiod and constant temperature conditions. At 1- to 2-month intervals, testicular width was measured by laparotomy, and blood samples were taken for analysis of plasma luteinizing hormone (LH). Most of the control birds went through at least one circannual cycle of testicular width and plasma LH concentration. In the castrates, a similar proportion of birds went through circannual LH cycles with periods indistinguishable from those of the controls. It is concluded that the testes and their hormones are not essential components of the mechanism that generates circannual gonadal cycles in male European starlings. PMID- 2979663 TI - Entrainment of duodenal activity to periodic feeding. AB - The electrical activity of the antrum and proximal duodenum was recorded simultaneously with bar pressing for food in various feeding conditions. During restricted feeding (RF; food available from 1200 to 1600 hr; lights-on from 0800 to 2000 h), large-amplitude slow potentials of duodenal origin were observed 3-5 hr preceding food access. These potentials occurred in short trains at irregular intervals of approximately 1 min and appeared to be associated with the active phase of the migrating motility complex. At least 8-10 days on RF were required before these irregular contractions (ICs) reliably preceded food access. The distribution of ICs showed a circadian variation during food deprivation, and transients lasting 4-6 days were observed in response to 6- or 8-hr phase delays of food access. These observations suggest that ICs are modulated by a circadian mechanism. On the average, the increase in ICs occurred about 2 hr earlier than bar pressing in anticipation of food, and there was no specific relation between individual ICs and bar pressing. However, bar pressing frequently occurred 0.5 min or longer after an increase in electromyogram (EMG) amplitude. Unreinforced bar pressing was never observed during a quiescent EMG. PMID- 2979664 TI - Entrainment of the circadian activity rhythm to the light-dark cycle can be altered by a short-acting benzodiazepine, triazolam. AB - The circadian rhythm of locomotor activity in hamsters maintained in either constant darkness or constant light can be phase-shifted by a single injection of the short-acting benzodiazepine, triazolam. These results suggest that treatment with triazolam may also alter the entrainment pattern of circadian rhythms in animals that are synchronized to a light-dark (LD) cycle. To test this hypothesis, hamsters maintained on an LD 6:18 light cycle received daily injections of triazolam (or vehicle) for 10-12 days, and any subsequent effects on the phase relationship between the onset of activity and the LD cycle were determined. Daily injections of triazolam (but not vehicle) induced pronounced advances or delays in the phase relationship between the entrained activity rhythm and the LD cycle; the direction of the shift was dependent on the time of the injection. Taken together with data from previous studies, these results suggest that triazolam, and perhaps other short-acting benzodiazepines, can be used to manipulate the mammalian circadian clock under a variety of experimental conditions. PMID- 2979665 TI - Circadian organization and photoreception in an Australian dasyurid marsupial (Sminthopsis macroura). AB - Much is known about the formal properties of circadian rhythm regulation and the physiological substrates underlying rhythmicity in nocturnal rodents, but relatively few studies have addressed circadian rhythm regulation in other mammalian taxonomic groups. In this study, some formal and functional aspects of circadian organization in a nocturnal dasyurid marsupial, the stripe-faced dunnart (Sminthopsis macroura), were analyzed. To determine phasic responses to discrete pulses of light, dunnarts were placed in constant darkness (DD) and were periodically administered pulses of bright light at different times of the animals' circadian day. Analysis of phase shifts in response to light indicated a phase response curve that was similar to responses observed in nocturnal rodents. To determine the possibility of extraretinal photoreception mediating photic entrainment, dunnarts were anesthetized and orbitally enucleated while maintained in a light-dark regimen (LD 14:10). All blinded dunnarts free-ran with periods (tau) that were similar to those observed in DD, indicating that entrainment is mediated through ocular photoreception. However, the data also indicated a decrease in activity in blind dunnarts during the last 3-5 hr of the dark phase, raising the possibility of some retention of photoreceptive capacities. PMID- 2979666 TI - The Paramecium circadian behavioral rhythm: light phase response curves and entrainment. AB - The population of a ciliate protozoan, Paramecium multimicronucleatum, exhibits a circadian rhythm as measured by the number of the cells traversing an observation point ("traverse frequency," or TF). The present study examined phase shifting of the TF rhythm by administering 2-hr light pulses at different phases of the circadian cycle to cultures free-running in constant darkness (DD). The results were summarized in a phase response curve (PRC), categorized as Type 1. This PRC indicated a relatively narrow phase zone insensitive to the light pulse ("dead zone"). Entrainment of the rhythm to light pulses repeated at 24-hr intervals was also examined, and it was found that the rhythm gradually reached a steady state, following several transient cycles, with the pulses falling at a phase corresponding to the narrow dead zone. Such a steady-state rhythm, with a minimum at approximately 3 hr after the pulse and a maximum at approximately 12 hr after the pulse, was mathematically simulated by superimposing a response function to the pulse on a sinusoidal function representative of the free-running rhythm in DD. PMID- 2979667 TI - Vertebrate circadian and photoperiodic systems: role of the pineal gland and melatonin. PMID- 2979668 TI - The mathematical structure of the human sleep-wake cycle. PMID- 2979669 TI - Response to the articles "Consultation-liaison psychiatry and the teaching of ethics," by J. R. McCartney and "Consultation-liaison psychiatry and clinical ethics," by J. R. Hayes. PMID- 2979670 TI - "Ethicogenesis": Response to the articles "Consultation-liaison psychiatry and the teaching of ethics," by J. R. McCartney, and "Consultation-liaison psychiatry and clinical ethics," by J. R. Hayes. PMID- 2979671 TI - Review: lipoxygenase inhibitors and the gut. AB - Leukotriene synthesis is influenced by several drugs currently in use for the treatment of alimentary disease, including the corticosteroids, sulphasalazine and mesalazine. However, the use of selective lipoxygenase inhibitors in human gastrointestinal disease has not been investigated. The complexity of eicosanoid metabolism, and the incomplete knowledge of roles played by each metabolite in each tissue and disease condition, make rational pharmacological manipulation of arachidonate metabolism difficult. However, lipoxygenase inhibitors show promise in animal models of inflammation, including hepatitis, and studies in vitro suggest that therapeutic benefits may be achieved using inhibitors of leukotriene synthesis in other inflammatory disorders. PMID- 2979672 TI - Evaluation of an enteric-coated delayed-release 5-aminosalicylic acid tablet in patients with inflammatory bowel disease. AB - Gastrointestinal transit of an enteric coated delayed release 5-aminosalicylic acid tablet radiolabelled with 111indium has been monitored in a total of 13 patients with Crohn's disease and ulcerative colitis. More than 70% of the tablets disintegrated in the small intestine, on average 3.2 hours after emptying from the stomach. Dispersed preparation was detected in the proximal colon of all the patients, except one with an ileostomy. Mean peak plasma concentrations of 5 aminosalicylic acid and its metabolite acetyl-5-aminosalicylic acid occurred 3-4 hours after gastric emptying. The tablets provide a reliable means of drug delivery to the ileum and proximal colon. PMID- 2979673 TI - Comparison of two antimuscarinic drugs, pirenzepine and propantheline, on gastric acid secretion, serum gastrin concentration, salivary flow and heart rate in patients with duodenal ulcer disease. AB - Effects of orally-administered pirenzepine and propantheline bromide on food stimulated gastric acid secretion, serum gastrin concentration, salivary flow and heart rate were compared in 10 duodenal ulcer patients in a placebo-controlled, double-blind study. Pirenzepine inhibited acid secretion by 25, 36 and 44% at doses of 50, 100, and 150 mg, respectively, while propantheline inhibited acid secretion by 32 and 41% at doses of 15 and 45 mg, respectively. None of the doses of pirenzepine affected food-stimulated serum gastrin concentrations, whereas 45 mg propantheline increased serum gastrin concentration significantly above placebo control. Enhancement of gastrin release by propantheline was not due to its antisecretory effect since intragastric pH after the meal was held constant at 5.0 by intragastric titration in vivo. Pirenzepine had no significant effect on heart rate and little or no inhibitory effect on salivary volume, depending on the dose administered. By contrast, both doses of propantheline increased heart rate and reduced salivary volume significantly (P less than 0.05). Thus, pirenzepine and propantheline in the doses administered inhibited acid secretion to approximately the same extent but pirenzepine had fewer effects on other organs. PMID- 2979674 TI - The effect of oral cisapride on colonic transit. AB - A prospective double-blind cross-over trial of oral cisapride 10 mg and placebo was performed to determine the effects of cisapride on the transit of colonic contents in normal humans. Six male volunteers were studied twice using colonic transit scintigraphy. After passing a tube to the caecum, 50 mu Ci of 111Indium diethylene triamine pentaacetic acid were instilled into the bowel lumen. The movement of radiolabelled material was followed using a gamma camera interfaced to a digital computer. Cisapride decreased the half-emptying of the caecum and ascending colon from 1.68 +/- 0.4 hours to 0.72 +/- 0.15 hours (P less than 0.05). The total colon half-emptying time was reduced from 38.5 +/- 7.2 hours to 11.1 +/- 2.9 hours on cisapride (P less than 0.05). Cisapride accelerated transit through the transverse colon, but not the descending colon. The progression of the geometric centre was faster during cisapride administration than with placebo (P less than 0.05). The number of bowel movements 48-hours-1 increased after cisapride from 2.5 +/- 0.8 to 5.0 +/- 0.4 (P less than 0.05). This study demonstrates that cisapride has a marked prokinetic effect on colonic transit in normal subjects. Cisapride may be a useful agent in the treatment of constipation. PMID- 2979675 TI - Marked suppression of stimulated gastric acid and pepsin secretion by enisoprost, a new PGE1 analogue. AB - The gastric antisecretory effects of three different doses of enisoprost, a new synthetic PGE1 analogue, were compared with placebo and misoprostol in 20 healthy male volunteers. Enisoprost 100, 200 and 400 micrograms all significantly (P less than 0.0001; ANOVA) suppressed histamine-stimulated acid and pepsin output when compared with placebo or misoprostol 200 micrograms. Misoprostol produced a significant decrease of stimulated acid output when compared with placebo (P = 0.0012). The concentration of pepsin in gastric juice was significantly (P less than 0.0001) decreased by enisoprost at the commencement of histamine stimulation. This effect was short-lived, and was maximal with enisoprost 400 micrograms. There was a significant dose-response relationship for enisoprost for inhibition of stimulated acid output (P = 0.0065). Enisoprost was well tolerated, and no consistent drug-related adverse effects were detected. The profile of antisecretory effect of enisoprost, producing marked suppression of both acid and pepsin secretion independently, is unusual. This combination of activity along with any mucosal protective properties might be particularly effective in the treatment of peptic ulcer disease. PMID- 2979676 TI - The effect of a long-acting somatostatin analogue (SMS 201-995) on intermediary metabolism and gut hormones after a test meal in normal subjects. AB - SMS 201-995 is an octapeptide analogue of somatostatin. The effect of a single subcutaneous (s.c.) injection of 50 micrograms SMS 201-995 on post-prandial intermediary metabolism was investigated in normal subjects. In spite of a long lasting post-prandial suppression of insulin secretion, there were no significant changes in the plasma concentration of alanine, glycerol, 3-OH-butyrate or lactate. However, SMS 201-995 impairs carbohydrate tolerance, probably due to inhibition of insulin secretion. Basal and post-prandial plasma concentrations of the gut regulatory peptides pancreatic glucagon, motilin, pancreatic polypeptide, gastric inhibitory polypeptide, enteroglucagon, gastrin and peptide YY were suppressed up to 5 hours after subcutaneous administration of a single dose of SMS 201-995. PMID- 2979677 TI - Idiopathic slow-transit constipation: whole gut transit times, measured by a new simplified method, are not shortened by opioid antagonists. AB - A simplified method for measuring mean whole gut transit time has been validated which avoids stool collection and is suitable for out-patients. Radio-opaque markers are swallowed daily for 14 days and the mean whole gut transit time is derived directly from an abdominal X-ray. Using this new technique, neither nalmefene (in a double-blind study of six patients) nor naloxone (in an open study of four patients) shortened mean whole gut transit time in idiopathic slow transit constipation. PMID- 2979678 TI - Review: first-pass metabolism by the gastrointestinal mucosa. AB - The bioavailability of orally administered drugs may be reduced due to presystemic elimination. The first-pass effect can occur in the gastrointestinal tract, the liver and lung. Although the liver is the main drug metabolizing organ in the body, the gut wall can play an important role in the first-pass metabolism of certain drugs. Both phase I (preconjugation) and phase II (conjugation) reactions have been described. However, while the oxidative metabolic capacity of the intestinal mucosa is considerably smaller than that of the liver, the activity of conjugation reactions in the gut may be close to that of the liver, and in some cases may exceed it. Sulphate conjugation is particularly important for steriod hormones such as ethinyloestradiol, and for the beta-adrenoceptor stimulants isoprenaline and isoetharine. Glucuronidation has been demonstrated to occur in man for morphine, paracetamol and oestrogens. Significant drug--drug interactions have been described involving drugs undergoing sulphate conjugation. The study of intestinal metabolism in vivo is difficult in man since direct methods (for example, hepatic portal vein catheterization) is justified in only a small number of patients. Therefore, much of our present understanding has been derived from various in-vitro studies involving intestinal sheets, mucosal biopsies, isolated enterocytes and microsomal preparations. PMID- 2979679 TI - Review: gut sterilization in inflammatory bowel disease. AB - Achieving a substantial reduction in the bacterial flora of the gut is a theoretically attractive means of treating inflammatory bowel disease, particularly colonic disease. There are practical difficulties in obtaining a sustained reduction of the colonic bacterial count, and the potential role of such a treatment regimen is therefore the initiation of remission. The data in the literature supporting such a suggestion are anecdotal, and a controlled study is indicated. PMID- 2979680 TI - Ranitidine or cimetidine for ulcer healing? PMID- 2979681 TI - Cimetidine decreases aspirin-induced gastric mucosal damage in humans. AB - Aspirin induces gastric mucosal damage in animals and humans. The purpose of this study was to examine whether cimetidine protects the human gastric mucosa from acute aspirin-induced damage. Eight healthy subjects were studied on 4 separate days. Cimetidine, 400 mg, or placebo was given orally 1 hour before initial endoscopy. The stomach was isolated and atropine given to suppress basal acid secretion. Each study consisted of four 15 min periods during which an acidic test solution was instilled into the stomach. During the second period only, either aspirin (1300 mg, 36 mmol) or control for aspirin (36 mmol HCl) was added to the test solution. Ion fluxes and gastric mucosal potential difference were measured, and endoscopy performed following each test. After placebo, aspirin significantly altered hydrogen ion flux and potential difference versus basal and control. Cimetidine decreased the damaging effect of aspirin. Endoscopic scores increased after aspirin plus placebo, whereas they remained unchanged after aspirin plus cimetidine. Therefore, cimetidine decreased aspirin-induced gastric mucosal damage in humans. As gastric acidity was identical during all studies, the effect of cimetidine was independent of gastric acid secretion. PMID- 2979683 TI - A comparative study of the effect of cimetidine and ranitidine on the rate of gastric emptying of liquid and solid test meals in man. AB - Paired studies were carried out on 18 healthy male volunteers (20.9 +/- 1.9 years; mean +/- S.D.) to compare the effect of oral doses of the H2-receptor antagonists, ranitidine and cimetidine, on the rate of gastric emptying of radiolabelled solid and liquid test meals. Oral administration of ranitidine 300 mg accelerated the emptying of a liquid meal from the stomach, but it had no significant effect on the rate of emptying of a solid meal. Oral administration of either 400 or 800 mg cimetidine did not alter the rate of emptying of either the liquid or the solid meals from the stomach. PMID- 2979682 TI - Prednisolone absorption in inflammatory bowel disease: correlation with anatomical site and extent. AB - Prednisolone absorption was studied in 13 normal subjects, eight patients with ulcerative colitis and 21 patients with Crohn's disease, by measuring plasma levels after a single oral dose. Absorption of the drug was delayed in all patient groups. The peak plasma level of the drug was lower in patients with extensive small bowel Crohn's disease. Patients in this category may need higher doses of oral prednisolone than other patients with inflammatory bowel disease. PMID- 2979684 TI - Comparison of the effectiveness of midazolam and diazepam in lipid emulsion as sedation during upper gastrointestinal endoscopy. AB - In a study of 101 patients undergoing upper gastrointestinal endoscopy, 90% of patients had complete amnesia for the procedure after intravenous midazolam (average dose 10 mg), but only 61% had complete amnesia after intravenous diazepam in lipid emulsion (average dose 18.4 mg) (P = 0.0006). However, when assessed by two different tests, recovery within the first hour was significantly more rapid after diazepam (P less than 0.0001). Prolonged sedation (over 20 hours after injection) was reported occasionally by patients who had received either drug. Thus, as with patients who have been sedated with diazepam, those who have been sedated with midazolam should also be advised to avoid driving or operating machinery for at least 24 hours after injection. PMID- 2979685 TI - Effect of two potent calmodulin antagonists on calcium transport of brush border and basolateral vesicles from human duodenum. AB - In the present in-vitro study we investigated the possible role of the calmodulin antagonistic drugs loperamide and calmidazolium in the regulation of transepithelial Ca2+ transport of human duodenum. Brush border membrane vesicles and basolateral membrane vesicles were simultaneously prepared from surgically resected pieces of morphologically intact human duodenum with a modified Percoll gradient centrifugation method. Brush border and basolateral membrane vesicles were characterized using enzyme marker analysis and electron microscopy: alkaline phosphatase was enriched 20-fold in brush border membrane vesicles, whereas [Na+ + K+]-stimulated adenosine triphosphatase was enriched 15-fold in basolateral membrane vesicles. Calmodulin activity was determined by a specific radioimmunoassay after solubilizing brush border and basolateral membrane vesicles in 1% Triton X-100. In basolateral membrane vesicles, we found no calmodulin activity. In brush border membrane vesicles calmodulin activity was impaired by 50% after pre-incubation with loperamide or calmidazolium. We measured calcium, sodium, D-glucose and D-mannitol uptake with a rapid filtration technique. Before the transport experiments, brush border and basolateral membrane vesicles were pre-incubated with 5 microM loperamide or 5 microM calmidazolium for 60 min at 5 degrees C. In drug-pretreated, brush border membrane vesicles calcium uptake was significantly reduced after 1 min incubation (-25% +/- 5%, P less than 0.05); this effect was completely reversed in the presence of 5 microM calmodulin. In basolateral membrane vesicles, we found two Ca2+ transport systems: (1) Na+/Ca2+ exchange and (2) ATP-dependent Ca2+ transport. In basolateral membrane vesicles loperamide had no effect. Calmidazolium had no effect on Na+/Ca2+ exchange, but significantly inhibited ATP dependent Ca2+ transport. This effect could not be reversed by calmodulin. PMID- 2979686 TI - Review: pancreatic enzyme replacement--applied physiology and pharmacology. AB - The treatment of pancreatic steatorrhoea can often be improved by attention to the pathophysiological and pharmacological principles involved. Factors influencing the efficacy of pancreatic enzyme replacement include physiological characteristics of the individual patient and pharmaceutical characteristics of the supplement. Different patients may be best suited by different preparations and there is no overall 'best buy'. The new enteric-coated microsphere formulations are often most effective, but also more expensive than conventional preparations. Adjunctive H2-blockade can help appropriately selected patients with resistant steatorrhoea. Knowledge of the underlying cause may guide the choice of supplement and its dose, but trials of different regimens may prove necessary. Successful management, particularly of malnourished patients, involves optimization of dietary fat intake in addition to enzyme replacement. PMID- 2979687 TI - Review: acute human models of gastric mucosal injury. AB - The use of acute human models of gastric mucosal injury has been stimulated by a need to understand more fully the problems of non-steroidal anti-inflammatory drugs but such models have other applications. None is ideal and they all share certain drawbacks. For none of them has a precise relationship to clinical events been established and they have all tended to be employed on a population of young healthy subjects who are not those at greatest clinical risk. Of individual methods mucosal potential difference is an indirect measure which is too often affected by other influences to be acceptable as a measure of mucosal injury when used alone, although it has some value as an adjunct to other measurements. Assay of DNA in gastric washings is a suitable technique for quantifying desquamation of gastric epithelial cells occurring in response to acute injury; on present evidence its significance is much more difficult to assess in the context of continuing challenge over several days. By contrast, measurement of microbleeding is more suitable for quantifying injury over several days of NSAID ingestion; little bleeding is recorded with a single acute challenge. Endoscopy can demonstrate macroscopic lesions which result from mucosal injury--injury which is quantified more easily and sensitively by measurements of cellular exfoliation or bleeding. Paradoxically, endoscopy's strength has been to underline the scientific weakness of acute models because it shows that it is rare for ulcers, which are the lesions of clinical concern to develop in these studies. PMID- 2979688 TI - Contractility of human gallbladder muscle in vitro. AB - This study examined the effects of transmural nerve stimulation, acetylcholine, adrenoceptor agonists and several peptides on the contractility of strips of human gallbladder in vitro. Acetylcholine caused concentration-related contractions of the tissues and the sensitivity to acetylcholine was similar in gallbladders with mild and severe chronic cholecystitis. Noradrenaline and adrenaline relaxed gallbladder strips, probably via beta 2-adrenoceptor stimulation. Transmural nerve stimulation always caused contractions, but in the presence of atropine inhibitory responses were demonstrable and these were antagonized by propranolol. There was no evidence of non-adrenergic inhibitory neural responses. Of the peptides tested, only cholecystokinin octapeptide (CCK OP), gastrin, pentagastrin, substance P and caerulein caused contractions. Responses to CCK-OP, gastrin and pentagastrin were antagonized by dibutyryl cyclic GMP. Hormones which had no effect upon human gallbladder strips included motilin, secretin, bombesin, neurotensin, glucagon, vasopressin, VIP and somatostatin. Considerable differences therefore exist between human tissues and those from experimental animals with respect to the direct actions of neural and hormonal stimuli on gallbladder contractility. PMID- 2979689 TI - Inhibition of prostanoid synthesis by human gastric mucosa. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) damage the gastric mucosa, and an important part of this effect is probably due to inhibition of prostaglandin synthesis. We have therefore studied various drugs for their ability to reduce prostaglandin and thromboxane formation by human isolated gastric mucosa. The overall relative potencies for inhibiting the endogenous production of PGE, 6 keto-PGF1 alpha and thromboxane B2 by mucosal pieces was generally: indomethacin = naproxen greater than ibuprofen greater than piroxicam; diflunisal, the prodrug sulindac, and the analgesic paracetamol usually had small or variable effects. This rank order was mainly similar to the inhibition of gastric microsomal PGE2 formation from exogenous arachidonic acid, the relative potencies being: indomethacin greater than naproxen greater than ibuprofen = piroxicam = diflunisal; again sulindac and paracetamol had little or no effect. The relative propensity of NSAIDs to cause gastric mucosal damage is controversial, but aspirin and indomethacin may be worst, and ibuprofen seems to be among the safest. Potency as an inhibitor of prostaglandin synthesis correlates better with the reported propensity for damage than does potency x dose. For reasons that are given in the discussion, this may indicate that gastric mucosal damage by NSAIDs with short or moderate half-lives is due largely to locally absorbed drug. Whereas inhibition of prostaglandin synthesis is probably the major cause of the damage, the simultaneous reduction of thromboxane formation might be advantageous for gastric mucosal integrity. Various implications arise from our hypotheses concerning the design of anti-inflammatory drugs. PMID- 2979690 TI - Effect of omeprazole on lower oesophageal function in normal subjects. AB - In a double-blind crossover study, lower oesophageal sphincter pressure (LOSP) and distal oesophageal motility were studied in eight healthy volunteers following a single intravenous dose of omeprazole or placebo. Lower oesophageal sphincter pressure was determined before and at intervals up to 120 min after intravenous administration of 40 mg omeprazole or placebo. No effects on LOSP or distal oesophageal motility were observed. PMID- 2979691 TI - Does Caved-S decrease the gastric ulcer relapse rate during maintenance treatment with ranitidine? AB - Ninety-five patients with healed gastric ulcers were treated with ranitidine 150 mg at night plus either Caved-S two tablets twice daily, or an identical placebo preparation. Treatment was continued for 2 years: the recurrence rate after 1 year was 12.5% for single therapy and 19% for combined therapy, and 30 and 40% respectively after 2 years (differences not significant). No serious side-effects were encountered, but patients found the combination therapy difficult to take, and it conferred no clinical advantage. In a 3 month follow-up after stopping maintenance therapy, 11% of the patients had a symptomatic ulcer recurrence. It is therefore recommended that elderly patients with gastric ulceration should remain on maintenance therapy for life. PMID- 2979692 TI - The effect of calcium channel blockade with nifedipine on splanchnic and systemic haemodynamics in cirrhosis. AB - The splanchnic and systemic haemodynamic effects of a single sublingual dose of nifedipine (slow calcium channel blocker) in nine patients with cirrhosis of the liver and portal hypertension were studied. Nifedipine produced a significant reduction in the mean arterial blood pressure (98 +/- 5.3 vs. 86 +/- 5 mmHg, P less than 0.05) but did not alter the mean heart rate, portal venous pressure or total liver blood flow. The systemic antihypertensive effect of nifedipine can be achieved without altering liver blood-flow in patients with chronic liver disease and portal hypertension. PMID- 2979693 TI - Effect of nicardipine on gastric acid secretion in man. AB - The effect of nicardipine on basal and pentagastrin-stimulated gastric acid secretion in normal volunteers was investigated. When compared with saline, an intravenous infusion of nicardipine caused a significant decrease in peak acid output (from 37.8 mmol hour-1 to 28.8 mmol hour-1; P = 0.04) and a small reduction in aspirate volume. Nicardipine had no significant action on basal acid output or volume of aspirate. Proteolytic activity in both the basal and stimulated periods was unaffected by nicardipine as were serum gastrin concentrations. Although calcium channel blocking agents are theoretically antisecretory the present study suggests they are unlikely to have clinically useful therapeutic actions. PMID- 2979694 TI - Is it really more difficult to treat prepyloric ulcers? AB - Prepyloric and duodenal ulcers have some common characteristics: gastric acid secretion is increased and there is an association with blood group O. Many, therefore, have considered prepyloric ulcers to be a variety of duodenal ulcer disease. From an anatomical point of view, however, prepyloric ulcers are clearly gastric ulcers. After proximal selective vagotomy, the recurrence rate is very high, amounting to more than 30% in 5 years; this is significantly higher than the rate for duodenal ulcers. Better results are obtained in prepyloric ulcers, if vagotomy is combined with a drainage procedure. In recent years, some evidence, primarily from Scandinavia, has accumulated indicating that prepyloric ulcers are more resistant to treatment with histamine H2-receptor antagonists than duodenal ulcers or ulcers located in other parts of the stomach. In addition, the recurrence rate is particularly high in prepyloric ulcers. One must, however, consider that not only have all of these studies included relatively small numbers of patients, but also the prepyloric ulcer healing rates in other studies were similar to those observed for both duodenal ulcers and ulcers located elsewhere in the stomach. Prospective studies with large numbers of patients are, therefore, necessary before a clear-cut conclusion can be reached. There are several reasons why prepyloric ulcers could be more resistant to treatment. Impaired gastric emptying, duodeno-gastric reflux or chronic gastritis, especially in conjunction with Campylobacter pylori infection, must be considered. At present, one can only speculate on the validity of any of these hypotheses. PMID- 2979695 TI - What is an intractable duodenal ulcer and how should it be managed? AB - An intractable duodenal ulcer is an ulcer that has not healed after 8 weeks of full-dose treatment with a modern anti-ulcer drug. Such ulcers are relatively rare--perhaps five in 100 patients will have intractable duodenal ulceration. The differential diagnosis includes non-compliance as a cause of continuing ulceration, hypersecretion of gastric acid or other rare causes of ulcers in the duodenum. Investigations should include biopsy of the continuing ulceration at endoscopy, measurement of fasting plasma gastrin and routine haematology and biochemistry profiles. An intractable duodenal ulcer can be treated either by using a powerful antisecretory regimen (high doses of ranitidine or famotidine or a conventional dose of omeprazole), by changing to tripotassium dicitrato bismuthate or, in exceptional circumstances, by extremely careful surgical intervention. PMID- 2979696 TI - When and why do ulcers bleed and what can be done about it? AB - This article reviews the pathophysiology and management of bleeding peptic ulcer. Ulcers bleed when and because they erode into a blood vessel, and bleed massively when they erode into a medium- or large-sized artery. Focal pathology at the bleeding point (such as arteritis, aneurysmal dilatation or recanalized thrombus) contributes to the timing and clinical pattern of ulcer bleeding. Big bleeds are probably associated with erosion into big arteries. The identification of a visible vessel in the floor of an ulcer that has recently bled is predictive of further bleeding, while the absence of a visible vessel or stigmata of recent haemorrhage is strongly predictive that further bleeding will not occur. Unfortunately, no conventional method of managing gastrointestinal bleeding from ulcers has been convincingly shown to be better than placebo in controlled clinical trials. The value of transfusion and surgery has never been tested in controlled trials, while many small studies of drug therapy, especially of H2 receptor blocking agents, and a few small studies of early surgery afford generally negative or equivocal results. There is some evidence that new physical methods such as lasers or bipolar probes applied at endoscopy are superior to placebo though negative trials have also been reported. Studies randomizing larger numbers of patients with bleeding ulcers are required if therapeutic benefit of any aspect of management is to be demonstrated or refuted. PMID- 2979697 TI - What are the current possibilities in treating peptic ulcer disease? AB - There are two major principles of ulcer therapy. Today, the most widely accepted drugs are those which substantially reduce aggressive factors (i.c. acid and pepsin), namely histamine H2-receptor antagonists, antimuscarinics and antacids. Less frequently applied are mucoprotective agents like colloidal bismuth compounds and sucralfate. Prostaglandins both reduce acid secretion substantially and are believed to enhance mucosal resistance. Their anti-ulcer efficacy, however, is solely explicable by their antisecretory activity. Although mucosa strengthening agents and H2-receptor blockers have nearly identical healing rates, mucosa-strengthening agents have inconvenient dosage regimens (four times or twice daily) and are probably less effective in relieving pain. The same holds true for antacids. Prostaglandins, antimuscarinics and antacids have dose related side effects. In contrast, H2-receptor blockers are characterized by a clear mechanism of action, convenient dosage regimens, good tolerance and a low incidence of side-effects. H2-receptor antagonists are the most effective anti ulcer drugs presently available. PMID- 2979698 TI - The place of famotidine in anti-ulcer therapy. AB - Famotidine now presents physicians in the USA and many European countries with a third option when considering H2-antagonist therapy. A dose of 40 mg in the evening decreases nocturnal gastric acidity for 10-12 hours, leaving daytime stimulated acidity virtually unaffected. This single evening dosage regimen produces effective healing of gastric and duodenal ulceration; maintenance of healing can then be achieved satisfactorily with 20 mg in the evening. In extensive clinical studies, the adverse effect profile of famotidine is similar to placebo. Famotidine has no known drug interactions and there are no identified mechanisms by which it might be expected to produce them. Circulating plasma hormone concentrations in man are not affected by famotidine and no antiandrogenic properties have been observed in animal studies. Future potential uses of famotidine may include the treatment of haemorrhage from peptic ulcers and the healing of oesophageal ulceration. PMID- 2979699 TI - What is new in the epidemiology and pathogenesis of peptic ulcer? AB - The old dictum 'no acid--no ulcer' is no longer a sufficient explanation of the pathogenesis of ulcer disease. The real question is 'if acid--why ulcer?' Although acid remains predominant, some of the other factors influencing ulcerogenesis are nocturnal acid secretion, pepsin enzyme subspecies, the mucus layer, bicarbonate levels, prostaglandins, Campylobacter pylori infection, consumption of non-steroidal anti-inflammatory drugs, and smoking habits. Although the ulcer burden has been greatly reduced by the introduction of H2 receptor antagonists, complications such as bleeding and perforation remain a problem, especially in the elderly. Medical treatment, in the form of H2-receptor antagonists, is effective for many patients. PMID- 2979700 TI - Lessons from prolonged gastric pH monitoring. AB - Intragastric pH monitoring has shown that the distribution of acidity within the stomach is not homogeneous. Not only is it affected by meals but it also has a circadian rhythm in which nocturnal pH falls to very acid levels in normal subjects. Although results of pH monitoring are highly reproducible within individuals, considerable inter-individual variation has been shown. Duodenal ulcer patients do not appear to possess the normal buffering reaction to meals, but their night-time acidity is within the normal range. In these patients, antacids and pirenzepine have a small acid-neutralizing effect in the stomach; cimetidine is less potent than ranitidine and famotidine. Clinicians can choose between a single dose of either ranitidine or famotidine in the evening with dinner and a twice-daily regimen. PMID- 2979701 TI - Significance of Campylobacter pylori. AB - There is an explosion of interest in the role of Campylobacter pylori as a cause of active chronic gastritis. This curved spiraled microorganism can readily be detected within the mucus gel covering the stomach mucosa, especially in patients suffering from peptic ulcer disease or non-ulcer dyspepsia. To what extent this intriguing microorganism is causally related to peptic ulcer disease remains to be elucidated, but all the evidence which is available so far supports a pathogenetically important role. There appears to be a striking discordance between in-vitro sensitivity and in-vivo efficacy of antibiotic therapy. At present, the combination of colloidal bismuth subcitrate and amoxycillin or tinidazole appears most effective in temporary elimination of these microorganisms. PMID- 2979702 TI - Why are non-steroidal anti-inflammatory drugs important in peptic ulceration? AB - The pathogenetic role of non-steroidal anti-inflammatory drugs (NSAIDs) in peptic ulcer disease is reviewed, on the basis of available experimental and epidemiological knowledge. In addition, original clinical data are provided concerning the prophylactic and therapeutic role of the H2-receptor antagonists ranitidine and cimetidine, and colloidal bismuth subcitrate, in the treatment of NSAID-associated peptic lesions in rheumatic patients. PMID- 2979703 TI - When should endoscopy (or radiology) be used in dyspepsia and peptic ulcer disease? AB - It is now possible readily to investigate dyspeptic symptoms using either a double-contrast barium meal or upper gastrointestinal endoscopy. The accuracy of endoscopy makes it preferable for routine use. As oesophagogastro-duodenoscopy (OGD) is invasive, some risks (albeit very small) are involved. Moreover, this technique has some weaknesses. For these reasons, selection of patients is important. Organic disease is most likely to occur in older patients: anyone presenting with dyspepsia for the first time over the age of 40 years should be investigated automatically. In individuals under 40 years of age, organic disease is less common and some selection criteria should be applied to reduce the number of negative investigations. After age, smoking is probably the single most important adverse factor. For gastric ulcer, endoscopy with biopsy, repeated after a course of therapy, is routine, but for duodenal ulcer repeat examination need not normally be undertaken owing to the effectiveness of modern healing drugs. Methods for improving the diagnostic accuracy of patient histories and clinical examinations need to be developed in order to utilize diagnostic investigations more efficiently for the patients' benefit. PMID- 2979704 TI - Interaction of nod and exo Rhizobium meliloti in alfalfa nodulation. AB - Among the genes of Rhizobium meliloti SU47 that affect nitrogen-fixing symbiosis with alfalfa are nod genes, in which mutations block nodule induction, and exo genes, in which mutations allow nodule formation but block rhizobial exopolysaccharide production as well as nodule invasion and nitrogen fixation. To investigate whether an exo+ bacterium can "help" (that is, reverse the symbiotic defect of) an exo mutant in trans, we have coinoculated alfalfa with pairs of rhizobia of different genotypes. Coinoculant genotypes were chosen so that the exo+ helper strain was nif while the exo "indicator" strain was nif+, and thus any fixation observed was carried out by the exo coinoculant. We find that a nod exo+ coinoculant can help an exo mutant both to invade nodules and to fix nitrogen. However, a nod+ exo+ coinoculant cannot help an exo mutant: Few exo bacteria are recovered from nodules, some bacteroids differentiate into bizarre aberrant forms, and the nodules fail to fix nitrogen. In a triple coinoculation, the effect of nod+ helper supersedes that of nod helper. Implications of these results for interaction of nod and exo gene products are discussed. PMID- 2979705 TI - Site-specifically modified oligodeoxynucleotides as probes for the structural and biological effects of DNA-damaging agents. PMID- 2979706 TI - Structure of the single stable hemoglobin adduct formed by 4-aminobiphenyl in vivo. PMID- 2979707 TI - Identification of crotonaldehyde as a hepatic microsomal metabolite formed by alpha-hydroxylation of the carcinogen N-nitrosopyrrolidine. AB - Crotonaldehyde (2-butenal), which reacts with DNA and is mutagenic and carcinogenic, was identified as a hepatic microsomal metabolite of the hepatocarcinogen N-nitrosopyrrolidine. Incubation mixtures of N nitrosopyrrolidine, cofactors, and hepatic microsomes from Aroclor pretreated or control F344 rats were derivatized with (2,4-dinitrophenyl)hydrazine reagent and the resulting mixtures analyzed by high-performance liquid chromatography. Crotonaldehyde (2,4-dinitrophenyl)hydrazone was identified by its retention time in two different systems and by its ultraviolet and mass spectrum. The ratio of 4 hydroxybutyraldehyde, which has previously been identified as a metabolite of NPYR, to crotonaldehyde was 1.5-2 over a range of substrate concentrations. The approximate values of Km and nu max for crotonaldehyde were 5.8 mM and 0.6 nmol/min/mg of protein and for 4-hydroxybutyraldehyde 14.1 mM and 1.7 nmol/min/mg of protein, for substrate concentrations between 1 and 8 mM, with microsomes from Aroclor pretreated rats. The ratio of 4-hydroxybutyraldehyde to crotonaldehyde was 1.9 upon esterase-catalyzed solvolysis of alpha-acetoxy-N-nitrosopyrrolidine, a stable precursor to the initial product of N-nitrosopyrrolidine alpha hydroxylation. These results demonstrate that crotonaldehyde is formed upon metabolic alpha-hydroxylation of N-nitrosopyrrolidine and suggest that it may be involved in N-nitrosopyrrolidine-macromolecule interactions. PMID- 2979708 TI - Base-catalyzed decomposition of N-nitrosobis(2-oxopropyl)amine. AB - N-Nitrosobis(2-oxopropyl)amine, a potent carcinogen for the pancreas in Syrian hamsters, undergoes a facile, base-catalyzed, intramolecular aldol condensation to yield N-nitroso-5-hydroxy-5-methyl-3-piperidone. This cyclic nitrosamine then decomposes to yield 3-hydroxy-5-methylpyridine. PMID- 2979709 TI - Cytotoxicity and bioactivation mechanism of benzyl 2-chloro-1,1,2-trifluoroethyl sulfide and benzyl 1,2,3,4,4-pentachlorobuta-1,3-dienyl sulfide. AB - The metabolism and cytotoxicity of benzyl 1,2,3,4,4-pentachlorobuta-1,3-dienyl sulfide (1) and benzyl 2-chloro-1,1,2-trifluoroethyl sulfide (2) were studied as an alternative test of the hypothesis that the toxicity of the cysteine S conjugates S-(pentachlorobutadienyl)-L-cysteine and S-(2-chloro-1,1,2 trifluoroethyl)-L-cysteine is associated with their metabolism to unstable thiols; the expectation was that the benzyl sulfides 1 and 2 would undergo cytochrome P-450 dependent benzylic hydroxylation and that the intermediate hemimercaptals would eliminate unstable, cytotoxic thiols. This expectation was realized: 1 and 2 were cytotoxic in isolated rat hepatocytes. The cytotoxicity of 1 was greater in hepatocytes from phenobarbital-treated rats compared with control rats and in male than in female rats and was inhibited by carbon monoxide and 2-(N,N-diethylamino)ethyl 2,2-diphenylvalerate HCl (SKF 525-A). Benzyl sulfides 1 and 2 were metabolized to benzaldehyde by rat hepatic microsomal fractions and by a purified, reconstituted cytochrome P-450PB-B system. Benzaldehyde was not cytotoxic. These results provide support for the hypothesis that benzyl sulfides 1 and 2 and the corresponding cysteine S-conjugates yield unstable thiols, which may give rise to acylating agents or to stable, but toxic, terminal products that are responsible for the cytotoxic effects of the benzyl sulfides and cysteine S-conjugates. PMID- 2979710 TI - Covalent binding of N-hydroxy-N-acetyl-2-aminofluorene and N-hydroxy-N-glycolyl-2 aminofluorene to rat hepatocyte DNA: in vitro and cell-suspension studies. AB - Two 2-aminofluorene-derived hydroxamic acids that differ only in the nature of the N-acyl group were examined for their relative abilities to undergo covalent binding to nucleic acids. Studies of the bioactivation of N-hydroxy-N-acetyl-2 aminofluorene (N-OH-AAF) and N-hydroxy-N-glycolyl-2-aminofluorene (N-OH-GAF) were conducted with hepatocyte suspensions and subcellular fractions prepared from male Sprague-Dawley rats. Both hydroxamic acid substrates displayed equal binding to both DNA and RNA after incubations with hepatocyte suspensions. The extent of binding of each substrate was approximately the same for DNA and RNA. Investigations with subcellular fractions revealed some major differences between the probable mechanisms by which the two substrates were covalently bound to exogenous DNA. In agreement with the prior literature reports, N-OH-AAF was extensively bound to DNA through the action of cytosol enzymes, including both N,O-acyltransferase and sulfotransferase. The microsomal enzyme fraction also catalyzed binding to DNA, and this process was completely inhibited by paraoxon. The covalent binding of N-OH-GAF to DNA was catalyzed by cytosol enzymes to a significant extent only in the presence of 3'-phosphoadenosine-5'-phosphosulfate, which suggests the action of sulfotransferase. Covalent binding of N-OH-GAF to DNA was minimal through the action of cytosolic N,O-acyltransferase, which confirms our earlier observation that N-OH-GAF is a potent suicide inhibitor of this enzyme. The microsomal fraction catalyzed the binding of N-OH-GAF to DNA at a rate that was about twice that observed for N-OH-AAF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979711 TI - Cytochrome P-450 isozyme selectivity in the oxidation of acetaminophen. AB - Highly purified isozymes of cytochrome P-450 catalyzed the formation of 3 glutathion-S-ylacetaminophen (GS-APAP) and 3-hydroxyacetaminophen (3-OH-APAP) from acetaminophen (APAP). A major isozyme from untreated male rats (P-450UT-A) catalyzed the formation of ca. 2.0 nmol/nmol of P-450/10 min of 3-OH-APAP and approximately 7.2 nmol of GS-APAP/nmol of P-450/10 min. Antibodies specific for cytochrome P-450UT-A caused a decrease in the amounts of both metabolites produced in microsomal incubations. In contrast to these results, two other constitutive P-450 isozymes from rat liver, cytochrome P-450UT-F and the female specific isozyme P-450UT-I, produced less of both oxidative metabolites. Moreover, they produced significantly more of the catechol metabolite than the glutathione conjugate. These results are in accord with the observation that male rats are more susceptible to acetaminophen hepatotoxicity than female rats. Isozymes induced by phenobarbital also produced more of the catechol than the glutathione conjugate. Conversely, the major isozyme induced by beta naphthoflavone, cytochrome P-450 beta NF-B, produced a significantly greater amount of GS-APAP than 3-OH-APAP. When comparison was made to a major phenobarbital inducible form (cytochrome P-450PB-B) a definite isozyme specificity for the formation of the two metabolites was seen. The catechol was formed at rates of 2.21 and 0.53 nmol/nmol of P-450/10 min by cytochromes P-450PB B and P-450 beta NF-B, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979712 TI - Identification of adducts formed by reaction of guanine nucleosides with malondialdehyde and structurally related aldehydes. AB - Malondialdehyde and a series of acrolein derivatives substituted in the beta position with good leaving groups react with guanine and guanine nucleosides to form two different types of adducts. The reaction with guanosine is typical. One adduct exhibits ultraviolet absorbance maxima at 253, 319, and 348 nm and is fluorescent. Its NMR spectrum exhibits three new aromatic proton resonances derived from malondialdehyde. The mass spectrum exhibits an M + 1 at 320. The spectroscopic properties are consistent with the structure, 3-beta-D-erythro pentofuranosyl-pyrimido[1,2-alpha]purin-10(3H)-one (PyP-ribose). The second guanosine adduct is an equal mixture of diastereomers that exhibit ultraviolet maxima at 217 and 244 nm and mirror image circular dichroism spectra. The NMR spectrum and mass spectrum (M + 1 = 392) indicate the addition of two molecules of MDA to one molecule of guanosine. Two-dimensional NMR (COSY) analysis reveals the presence of propano and enal functionalities. The spectroscopic and chemical properties suggest an oxadiazabicyclo[3.3.1]nonene structure that is confirmed by X-ray crystallography. Comparison of the deoxyguanosine adducts of malondialdehyde to those of the structurally related carbonyl compounds, methyl glyoxal and acrolein, provides a structural basis to explain the unique ability of malondialdehyde to induce frameshift mutations in bacterial mutagenesis systems. PMID- 2979713 TI - Fluorescence line narrowing-nonphotochemical hole burning spectrometry: femtomole detection and high selectivity for intact DNA-PAH adducts. AB - A new fluorescence line narrowing (FLN) apparatus is described and evaluated through experiments on intact DNA-PAH (polycyclic aromatic hydrocarbon) and globin-PAH adducts, as well as polar PAH metabolites. A detection limit of approximately 3 modified bases in 10(8) for a DNA adduct formed with a diol epoxide of benzo[a]pyrene (BPDE-DNA) is reported for 20 micrograms of DNA at a spectral resolution of approximately 8 cm-1. The methodology employed avoids or minimizes spectral degradation and loss of sensitivity due to photooxidation and nonphotochemical hole burning (NPHB). A new double selection technique that employs both FLN and NPHB is described and found to lead to a significant improvement in selectivity over that obtained with conventional FLN. PMID- 2979714 TI - Carboxylesterase isoenzyme specific deacylation of diacetoxyscirpenol (anguidine). AB - The deacylation reaction of diacetoxyscirpenol (DAS), a trichothecene mycotoxin, was studied by using five carboxylesterase isoenzymes isolated from mouse liver microsomes. The simultaneous isolation of the five microsomal carboxylesterase isoenzymes was accomplished by chromatofocusing. DAS serves as a unique substrate since both acetyl groups are accessible for hydrolysis, providing an opportunity to investigate the regioselective hydrolysis of DAS with the various isoenzymes. A novel basic carboxylesterase isoenzyme, pI 8.4-8.2, was isolated. This isoenzyme exhibits strict regioselectivity toward DAS hydrolysis; only the acetyl group at C-4, but not that at C-15, is hydrolyzed. Moreover, this is the major isoenzyme responsible for the deacylation of DAS in mouse hepatic microsomes. The rate constant determined for deacylation of DAS at C-4 for this isoenzyme is 130 1000 times greater than that of the other carboxylesterases. This isoenzyme was also the most efficient for the hydrolysis of 4-monoacetoxyscirpenediol with a rate constant 30-100 times greater than that of the other isoenzymes. PMID- 2979715 TI - Metabolism of procainamide to a hydroxylamine by human neutrophils and mononuclear leukocytes. AB - The chronic use of procainamide is associated with a high incidence of drug induced lupus and also agranulocytosis. We have previously demonstrated that procainamide is metabolized in the liver to reactive hydroxylamine (PAHA) and nitroso (nitroso-PA) metabolites which covalently bind to protein and are toxic to lymphocytes. We proposed that these metabolites were responsible for the toxicities of procainamide. However, PAHA and nitroso-PA do not appear to escape the liver in significant concentrations. In this paper we describe the metabolism of procainamide to a reactive hydroxylamine by neutrophils and mononuclear leukocytes. Such metabolism only occurs if the cells have been stimulated to have a respiratory burst. These observations have obvious possible implications for the mechanism of procainamide-induced agranulocytosis (formation of a reactive metabolite by neutrophils) and procainamide-induced lupus (formation of a reactive metabolite by monocytes). The metabolism of drugs to reactive metabolites by monocytes may be a general mechanism for hypersensitivity reactions because monocytes play a key role in the processing of antigen and stimulation of antibody synthesis. PMID- 2979716 TI - Mitochondrial reduction of the carcinogen chromate: formation of chromium(V). AB - Incubation of chromate with isolated rat liver mitochondria in vitro resulted in the uptake and reduction of chromium(VI), as well as the formation of chromium(V) species. Chromate was rapidly taken up and reduced by intact mitochondria. The rate of reduction of chromate by intact mitochondria was increased upon addition of succinate or malate plus glutamate, substrates for the electron-transport chain, but was decreased upon addition of cyanide, an inhibitor of the electron transport chain. Incubation of chromate with mitochondria in the presence or absence of malate, glutamate, and succinate resulted in a steady increase in the level of chromium(V) over time. The extent of chromium(V) formation was increased upon addition of malate, glutamate, and succinate but was inhibited upon addition of the electron-transport chain inhibitors, antimycin, cyanide, or rotenone, to whole mitochondria. High levels of glutamate plus malate inhibited chromium(V) formation; however, high concentrations of succinate or sulfate had no effect. These studies suggest that the chromate-reductase activity in mitochondria is due to the electron-transport chain as well as other mitochondrial reducing systems which are insensitive to inhibitors of the electron-transport chain. Since chromium(VI) is effectively metabolized by mitochondria in vitro and chromium(V) "reactive intermediates" are formed in the process, mitochondria may play a role in chromium(VI) carcinogenesis. PMID- 2979717 TI - Identification of a novel dihydroxy metabolite of aflatoxin B1 produced in vitro and in vivo in rats and mice. AB - HPLC analysis of bile obtained from rats given aflatoxin B1 (AFB) demonstrates the presence of numerous polar metabolites. The glutathione conjugate derived from AFB 8,9-epoxide and the glucuronide conjugate of aflatoxin P1 (AFP) have previously been identified as the two major polar metabolites. The most polar peak present in bile from AFB-treated rats is converted to a less polar peak upon incubation with beta-glucuronidase, which has a parent ion m/e of 314 amu. Treatment of this aglycon with diazomethane produced a product which cochromatographs with aflatoxin M1 (AFM). From these data it is concluded that the most polar peak in bile from AFB-treated rats is the glucuronide conjugate of 4,9a-dihydroxyaflatoxin B1. This dihydroxy AFB metabolite was produced in vitro in mouse microsomal incubations, and time-course studies of its production suggest that it is largely formed by 9a-hydroxylation of AFP, although some may be formed by 4-O-demethylation of AFM. Direct incubation of AFP and AFM with mouse microsomes confirmed that this metabolite can be formed from both AFP and AFM. An HPLC method is described which is capable of base line resolution of this novel dihydroxyaflatoxin metabolite and eight other hydroxylated metabolites of AFB, as well as the glutathione conjugate of AFB 8,9-epoxide. PMID- 2979718 TI - Microsomal metabolism of 4-(N,N-diacetylamino)benzo[a]pyrene: a potent mutagenic arylamide derived from a carcinogenic polycyclic aromatic hydrocarbon. AB - 4-(N,N-Diacetylamino)benzo[a]pyrene, a potent mutagen, is derived from a carcinogenic polycyclic aromatic hydrocarbon, benzo[a]pyrene. Metabolism of this compound by rat liver microsomes was studied. Metabolites were separated by reversed-phase high-performance liquid chromatography and were identified by analysis of their UV-vis absorption, mass, and proton nuclear magnetic resonance spectral data. Seven metabolites were identified, namely, the 9-phenol, 1,9 hydroquinone, and trans-9,10-dihydrodiol of 4-(N,N-diacetylamino)benzo[a]pyrene, 4-(N-acetylamino)benzo[a]pyrene, and the 5-phenol, 5,9-hydroquinone, and trans 9,10-dihydrodiol of 4-(N-acetylamino)benzo[a]pyrene. Comparison of these results with those of metabolism of benzo[a]pyrene indicates that the N,N-diacetylamino substitutent at the 4-carbon of benzo[a]pyrene inhibits metabolism at the peri position (3-carbon) and positions (6-, 7-, and 8-carbons) remote from the substituent. The results also indicate that while 4-(N,N-diacetylamino) benzo[a]pyrene is a substrate of the rat liver microsomal deacetylase, the formed 4-(N-acetylamino)benzo[a]pyrene apparently is not a substrate. PMID- 2979719 TI - A structure-activity relationship study of organophosphorus compounds. AB - Organophosphorus compounds have been shown to exhibit toxic behavior as insecticides, pesticides, and mammalicides. Soman and 21 related compounds were studied for possible structure-activity relationships. Computer-aided methods were used to generate a linear expression relating the activity (ln [1/LD50], rabbit I.V.) of the compounds to three structure-based descriptors (R = 0.96). Principal components regression and jackknife analysis were performed to assess the stability of the model. PMID- 2979720 TI - Metabolic studies on phencyclidine: characterization of a phencyclidine iminium ion metabolite. AB - Studies on the metabolic bioactivation of the psychotomimetic amine phencyclidine have been pursued through the characterization of a new metabolite which is formed via initial cytochrome P-450 catalyzed oxidation of the parent drug to the corresponding iminium species. CI mass spectrometric and diode array UV and 1H NMR spectral analyses provided evidence for the conjugated amino enone compound, 1-(1-phenylcyclohexyl)-2,3-dihydro-4-pyridone. Confirmation of the proposed structure was achieved by comparing the 1H NMR and high-resolution EI mass spectral properties of the metabolic isolate with the corresponding spectra of an authentic synthetic sample. Possible intermediates involved in the formation of the dihydropyridone metabolite from the phencyclidine iminium ion are discussed in terms of structural analogies to reactive intermediates formed in the bioactivation of the nigrostriatal toxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). PMID- 2979721 TI - Reactivity of perhydroxyl (HOO.) with 1,4-cyclohexadiene (model for allylic groups in biomembranes). AB - The electron-transfer reduction of molecular oxygen yields superoxide ion (O2.-), which reacts with proton sources to form HO2.. In water the latter species disproportionates via reaction with O2.- (kbi, 10(8) M-1 s-1) and itself (kd, 10(6) M-1 s-1). The rate constants (kd) for the homolytic disproportionation process (HO2. + HO2.----H2O2 + O2), which have been determined from stopped-flow spectrophotometric decay data for HO2. at 25 degrees C, are (1.7 +/- 0.5) x 10(4) M-1 s-1 in dimethyl sulfoxide (Me2SO), (5.3 +/- 0.5) x 10(4) M-1 s-1 in dimethylformamide (DMF), and approximately 10(7) M-1 s-1 in acetonitrile. With limiting fluxes of protons to control the rate of formation of HO2. from O2.-, the rate of decay of HO2. is enhanced by reaction with the allylic hydrogens of excess 1,4-cyclohexadiene (RH). On the basis of such data the apparent second order process (HO2. + RH----R. + H2O2) has a rate constant (kox) of (1.6 +/- 0.6) x 10(2) M-1 s-1. The reactivity of HO2. decreases as its solvation energy increases. PMID- 2979722 TI - Mechanism of drug-induced lupus. PMID- 2979723 TI - Effect of bilayer structure on the rates of reaction of ozone with polyunsaturated fatty acids in phosphatidylcholine liposomes. PMID- 2979724 TI - The effect of DNA sequence, ionic strength, and cationic DNA affinity binders on the methylation of DNA by N-methyl-N-nitrosourea. PMID- 2979725 TI - Synthesis and conformation of a dinucleoside monophosphate modified by aniline. AB - The modified dinucleoside monophosphate, N-[deoxycytidylyl-(3'-5')-guanosin-8 yl]aniline (dCprG-An) has been prepared by the phosphotriester synthesis approach, using suitably blocked derivatives of dCp and N-guanosin-8-ylaniline (rG-An). The latter compound was synthesized by a route that featured nucleophilic displacement by antiline upon an 8-bromoguanosine derivative. A number of attempts to prepare N-(deoxyguanosin-8-yl)aniline (dG-An) by electrophilic substitution, using activated aniline derivatives, failed. Nucleophilic substitution reactions of aniline with 8-bromodeoxyguanosine derivatives afforded only the base, N-guanin-8-ylaniline. The conformation of dCprG-An has been studied by CD, proton magnetic resonance, and minimized potential energy calculations. A flexible molecule with a mixture of conformers is indicated. Base-base stacked states predominate, in contrast to the case of a dimer containing 4-aminobiphenyl bound to the 8-position of guanine, where carcinogen-base stacked states are dominant. The mutagenic and carcinogenic activities of aniline are much less than those of many polycyclic aromatic amines. The diminished stacking ability of the aniline ring, as well as the weak electrophilic reactivity of activated aniline derivatives, may be a cause of this weak biological activity. PMID- 2979726 TI - Construction of an Escherichia coli vector containing the major DNA adduct of activated benzo[a]pyrene at a defined site. AB - The mutagenic and carcinogenic substance benzo[a]pyrene reacts with DNA following activation to its corresponding 7,8-diol 9,10-epoxide (BPDE), and the major DNA adduct (BP-N2-Gua) is formed when the C(10)-position of BPDE reacts with the N2 position of guanine. It is unknown if this adduct is a premutagenic lesion in vivo. Herein, the construction and characterization of an M13mp19-based, E. coli vector that contains BP-N2-Gua located in the unique PstI restriction endonuclease recognition site at nucleotide position 6249 in the (-)-strand is described (designated, BP-N2-Gua-M13mp19). First, the oligonucleotide 5'-TGCA-3' was reacted with BPDE and a product (5'-T(BP-N2)GCA-3') was isolated by HPLC that, when enzymatically digested to deoxynucleosides, yielded an adduct that comigrated on HPLC with an authentic BP-N2-Gua deoxynucleoside standard. Second, the 5'-hydroxyl group of 5'-T-(BP-N2)GCA-3' was phosphorylated with ATP and T4 polynucleotide kinase, and the product (5'-pT(BP-N2)GCA-3') was purified by HPLC. This product is stable when heated at 80 degrees C at both neutral and alkaline pH. Third, M13mp19 was manipulated such that the sequence 5'-pTGCA-3' was selectively removed from the (-)-strand in its unique PstI recognition site, and 5'-pT(BP-N2)GCA-3' was ligated into this gap with T4 DNA ligase and ATP. The product of this reaction (BP-N2-Gua-M13mp19) was shown to be insensitive to cleavage by PstI, which suggests that a modification is located in the PstI recognition site. The most likely modification is the adduct BP-N2-Gua. PMID- 2979727 TI - Chemistry of aminoiminomethanesulfinic and -sulfonic acids related to the toxicity of thioureas. AB - The reactions of aminoiminomethanesulfonic acid, phenylaminoiminomethanesulfonic acid, and N,N'-diphenylaminoiminomethanesulfonic acid in aqueous media at pH 7.4, 10, and 13-14 were investigated. At neutral pH hydrolysis to the corresponding urea was the major pathway for all three compounds. At higher pH phenylaminoiminomethanesulfonic acid reacted to give phenylcyanamide in nearly quantitative yield, while N,N'-diphenylaminoiminomethanesulfonic acid gave diphenylcarbodiimide which reacted further to give N,N'-diphenylurea. At pH 10 aminoiminomethanesulfonic acid reacted with itself, eventually giving N cyanoguanidine, while at pH 13-14, elimination to cyanamide predominated. The reactions of glycine with the aminoiminomethanesulfonic acids gave guanylated acetic acids as products. The rates of these nucleophilic substitutions of the sulfonic acid group of the aminoiminomethanesulfonic acids by the amino group of glycine decreased in the order aminoiminomethanesulfonic acid greater than phenylaminoiminomethanesulfonic acid greater than (2 methylphenyl)aminoiminomethanesulfonic acid greater than (2,6 dimethylphenyl)aminoiminomethanesulfonic acid. Higher relative rates of substitution of the aminoiminomethanesulfonic acids appear to be related to higher relative toxicities for the corresponding thioureas. PMID- 2979728 TI - Thioacylating intermediates as metabolites of S-(1,2-dichlorovinyl)-L-cysteine and S-(1,2,2-trichlorovinyl)-L-cysteine formed by cysteine conjugate beta-lyase. AB - The bioactivation mechanism of S-(1,2-dichlorovinyl)-L-cysteine (DCVC) and S (1,2,2-trichlorovinyl)-L-cysteine (TCVC) was studied with cysteine conjugate beta lyase (beta-lyase) from Salmonella typhimurium and with the pyridoxal phosphate model N-dodecylpyridoxal bromide (PL-Br) as catalysts and with GC/MS to identify the metabolites formed. PL-Br converted S-2-benzothiazolyl-L-cysteine to 2 mercaptobenzothiazole and S-benzyl-L-cysteine to benzyl mercaptan, demonstrating the ability of PL-Br to serve as a model for beta-lyase. PL-Br and bacterial beta lyase converted DCVC to chloroacetic acid and chlorothionoacetic acid and TCVC to dichloroacetic acid. Incubations of PL-Br with the S-conjugates in the presence of diethylamine resulted in the formation of N,N-diethylchlorothioacetamide from DCVC and of N,N-diethyldichlorothioacetamide from TCVC. Attempts to trap the enethiols, which are the expected initial products formed by beta-elimination, by reaction with methyl iodide in incubations with the beta-lyase model were not successful. The formation of thioacylating agents from the enethiols may contribute to the cytotoxic and mutagenic effects of DCVC and TCVC. PMID- 2979729 TI - Pyrrole oxidation and protein cross-linking as necessary steps in the development of gamma-diketone neuropathy. AB - It has been well documented that the gamma-diketone HD1 is the ultimate toxic metabolite of n-hexane. Furthermore, it has been shown that the pathogenetic mechanism by which HD exerts its neurotoxic effects is through binding to protein lysly residues and cyclization to pyrroles. The present study sought to determine whether the presence of pyrrole residues on NF1 proteins is sufficient to cause the NF-filled axonal swellings associated with n-hexane and other gamma-diketone neuropathies or whether pyrrole oxidation and protein cross-linking also have to occur in order for neurotoxicity to develop. We synthesized the HD analogue AcHD1 and assessed its rate of pyrrole formation in vitro, the ease of oxidation of its resulting pyrroles, and its ability to cross-link proteins in vitro. The in vivo effects of AcHD on rats were examined following daily ip1 injections. AcHD was found to have a rate of pyrrole formation comparable to that of the potent HD analogue DMHD1 at 35 degrees C. The pyrrole derived from AcHD was more resistant to oxidation than that derived from the neurotoxic compound HD. AcHD did not cross-link proteins in vitro. Pyrrole derivatives were demonstrated on hemoglobin isolated from animals treated with HD, DMHD, and AcHD. Cross-linked spectrin was detected in animals treated with HD and DMHD but not with AcHD. Rats receiving 0.1 or 0.25 mmol of AcHD/kg/day did not reach the end point of hindlimb paralysis observed in the gamma-diketone neuropathies, and the NF-filled axonal swellings seen following exposure to the neurotoxic gamma-diketones were not observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979730 TI - Metabolic studies on the nigrostriatal toxin MPTP and its MAO B generated dihydropyridinium metabolite MPDP+. AB - The metabolic fates of the nigrostriatal toxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) and its two-electron oxidation product, the 1-methyl-4 phenyl-2,3-dihydropyridinium species (MPDP+), have been examined in mouse brain and liver tissue fractions. Incubations of MPTP (50 microM and 1 mM) with mouse brain preparations result in the expected MAO B catalyzed formation of MPDP+. The four-electron oxidation product, the 1-methyl-4-phenyl-pyridinium species (MPP+), was the only other metabolite detected. The oxidation of 50 microM MPDP+ to MPP+ in the same preparations appears to be mediated by unidentified components present in membrane containing structures. The behavior of MPTP in mouse liver subcellular fractions is considerably more complex. NADPH-supplemented liver microsomes convert MPTP to the desmethyl and N-oxide metabolites. At high (1 mM) initial concentrations of MPTP there is evidence that NADPH-dependent, pargyline insensitive liver microsomal enzymes also catalyze the oxidation of MPTP to MPDP+. Incubations of MPDP+ with mouse liver preparations containing the cytosolic fraction led to the rapid disappearance of the substrate and the quantitative formation of a metabolic product with mass spectral and diode array UV characteristics expected for a lactam structure. Menadione, an inhibitor of the cytosolic enzyme aldehyde oxidase, inhibits the formation of this product. Unambiguous characterization of this metabolite as 1-methyl-4-phenyl-5,6-dihydro 2-pyridone was achieved by comparison of its high-resolution 1H NMR and high resolution EI mass spectra with the corresponding spectra of a synthetic standard.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979731 TI - Metabolic activation of valproic acid and drug-mediated hepatotoxicity. Role of the terminal olefin, 2-n-propyl-4-pentenoic acid. PMID- 2979732 TI - Aliphatic diketones: influence of dicarbonyl spacing on amine reactivity and toxicity. PMID- 2979733 TI - 2,2-Dialkyl-1,2-dihydroquinolines: cytochrome P-450 catalyzed N-alkylporphyrin formation, ferrochelatase inhibition, and induction of 5-aminolevulinic acid synthase activity. AB - Incubation of 2,4-diethyl-1,2-dihydro-2-methylquinoline (DMDQ) with hepatic microsomes from rats pretreated with phenobarbital, 3-methylcholanthrene, pregnenolone-16 alpha-carbonitrile, or dexamethasone results in minor loss of the cytochrome P-450 chromophore and accumulation of a hepatic pigment. The hepatic pigment consists of the four regioisomers of N-ethylprotoporphyrin IX and minor amounts of the corresponding N-methyl regioisomers. Exposure of chick embryo liver cells to DMDQ results in inhibition of their ferrochelatase activity, induction of their 5-aminolevulinic acid synthase activity, and accumulation of protoporphyrin IX. 1,2-Dihydro-2,2,4-trimethylquinoline (TMDQ) causes negligible loss of cytochrome P-450 in rat liver microsomes but in vivo still produces the four N-methylprotoporphyrin IX regioisomers in low yield. Furthermore, it inhibits ferrochelatase activity, elevates 5-aminolevulinic acid synthase activity, and causes protoporphyrin IX accumulation in cultured chick embryo hepatocytes. One-electron oxidation of the 2,2-dialkyl-1,2-dihydroquinolines to radical cations is postulated to result in N-alkylation of the prosthetic heme group of cytochrome P-450. The N-alkylprotoporphyrins IX thus formed are potent inhibitors of ferrochelatase. Inhibition of ferrochelatase causes the induction of 5-aminolevulinic acid synthase and the accumulation of protoporphyrin IX. Heme alkylation and ferrochelatase inhibition may be generally associated with substrates that are subject to cytochrome P-450 mediated oxidative extrusion of alkyl radicals. PMID- 2979734 TI - Characterization of 7,12-dimethylbenz[a]anthracene-adenine nucleoside adducts. AB - Chromatographic comparisons were made between radioactive adducts derived from the DNA of cells treated with [3H]7,12-dimethylbenz[a]anthracene and adducts derived from calf thymus DNA or nucleotides which had been treated in vitro with the synthetic syn 3,4-dihydrodiol 1,2-epoxide of this same carcinogen. This confirmed that three of the adducts formed in cells were derived from reaction of this particular dihydrodiol epoxide with deoxyadenosine while a fourth adduct was derived from its reaction with deoxyguanosine. After reaction of the dihydrodiol epoxide with polyadenylic acid, two ribonucleoside adducts were characterized by spectroscopic methods and were shown to have arisen from the cis opening of the epoxide ring at C1 by the amino group of adenine residues. PMID- 2979735 TI - N-glycolylhydroxamic acids: an improved synthetic method and the in situ generation and intramolecular rearrangement of N-acetoxy-N-glycolyl-2 aminofluorene. AB - A new and improved method for the synthesis of glycolylhydroxamic acids is described. The two-step method involves acylation of arylhydroxylamines with acetoxyacetyl chloride, followed by saponification of the ester bond to give the desired products. The conversion of hydroxamic acids to their thallous salts followed by subsequent acetylation with acetyl chloride to give N-acyloxy esters is described. During the course of this investigation, it was observed that the N acetoxy ester of an N-glycolylhydroxamic acid was highly unstable and underwent a novel O----O acyl migration. This facile rearrangement reaction was studied for the case of N-acetoxy-N-glycolyl-2-aminofluorene (N-OAc-GAF), which gave N hydroxy-N-(acetoxyacetyl)-2-aminofluorene (N-OH-AcAAF) as the sole product of this rearrangement. HPLC was used to investigate this reaction and included the assignment of an HPLC peak to be due to N-OAc-GAF. A competition study employed the amide N-glycolyl-2-aminofluorene (GAF) and demonstrated the absence of intermolecular transfer of the N,O-acyl (acetyl) group of N-OAc-GAF. The mechanism for the probable intramolecular rearrangement reaction is presented, along with a consideration of the possible significance it might have for the toxicity of glycolylhydroxamic acids. PMID- 2979736 TI - Metabolism of [14C]MPTP in mouse and monkey implicates MPP+, and not bound metabolites, as the operative neurotoxin. AB - The distribution, identification, and binding of [phenyl-14C]- or [methyl 14C]MPTP metabolites have been determined in brains of mouse and monkey exposed to toxic doses of MPTP. The distribution of radiolabeled metabolites was heterogeneous, with levels of MPP+ 1-100 mumol/L in dissected and homogenized monkey brain tissues. MPP+ constituted greater than 98% of all tissue radioactivity remaining at 1-3 days in the monkey and was identified in both cortical and striatal tissue. The relevance of the 2% of unextractable ("bound") radiolabeled metabolite was assessed in mouse brain by using pargyline or mazindol pretreatments which block dopamine depletion. The amount of binding increased rather than decreased when [phenyl-14C]MPTP was used along with pargyline or mazindol but was unchanged when [methyl-14C]MPTP was employed. This demonstrates that bound metabolites are inversely correlated to neurotoxicity as well as being N-demethylated. Two extractable metabolites, demethylated MPTP (PTP) and 1-methyl-4-phenyl-2-pyridone, were found at 30-min survival times in mouse brain and probably derive from peripheral metabolism of MPTP. At 4 h, mouse brain profiles of extractable metabolites resembled those from monkey brain, containing MPP+ as the predominant (greater than 90%) constituent. The similarity of MPP+ concentrations in mouse and monkey brain homogenates with those concentrations of MPP+ known to produce biological effects in vitro, along with the inverse relationship between bound metabolites and neurotoxicity, supports the intermediacy of MPP+ as the operative neurotoxin. PMID- 2979737 TI - Use of the Topliss scheme for the design of more effective chelating agents for cadmium decorporation. AB - The Topliss scheme, which designates a special series of substituents for use in sequence in the search for suitably substituted aromatic compounds with biological properties superior to the parent unsubstituted aromatic compound, has been applied to the design of new chelating agents for the decorporation of cadmium from its deposits in vivo. Substituted benzylamines have been used in the syntheses of the corresponding substituted sodium N-benzyl-D-glucamine-N carbodithioates, i.e., RC6H4CH2N(CS2Na)CH2(CHOH)4CH2OH (or RBGDTC): CF3BGDTC, R = 3-CF3; MeBGDTC, R = 4-CH3; MeOBGDTC, R = 4-OCH3; ClBGDTC, R = 4-Cl. Examination of the behavior of these compounds as agents for the mobilization of cadmium from its deposits in vivo shows that such substitution can have an appreciable effect on the organ selectivity for cadmium removal, as well as the efficacy. Of the four compounds suggested by the Topliss scheme for pilot studies, two were much more effective than the parent compound (BGDTC), and of these the best one, sodium N-(4-methoxybenzyl)-D-glucamine-N-carbodithioate (MeOBGDTC) reduced kidney cadmium levels to 30% and liver cadmium levels to 50% of those obtained with the parent compound. Sodium N-[3-(trifluoromethyl)benzyl]-D-glucamine-N carbodithioate (CF3BGDTC) was also found to be superior to the parent compound in reducing the liver cadmium levels, though it was equivalent in reducing renal cadmium levels. PMID- 2979738 TI - Leurosine biotransformations: an unusual ring-fission reaction catalyzed by peroxidase. AB - The dimeric Vinca alkaloid leurosine undergoes an unusual fission of the piperidine ring of the Iboga substructure when reacted with horseradish peroxidase and hydrogen peroxide. A preparative-scale oxidation of leurosine provided 15'-hydroxycatharinine, which was identified by infrared and proton and carbon-13 NMR spectroscopies and high-resolution mass spectrometry. A proposed pathway for the formation of 15'-hydroxycatharinine involves radical and iminium intermediates and cleavages of a putative diol. The enzymatic transformation product is 3 orders of magnitude less active than leurosine or vinblastine in vitro, in inhibiting the polymerization of tubulin. PMID- 2979739 TI - Metabolism and DNA binding of 1-nitropyrene and 1-nitrosopyrene in newborn mice. AB - This study was carried out in order to evaluate possible mechanisms responsible for tumor induction by 1-nitropyrene and to provide insights on the higher tumorigenicity of 1-nitrosopyrene than 1-nitropyrene in newborn mouse liver. The "ground mouse" technique was used to follow the development of the metabolism of 1-nitropyrene and 1-nitrosopyrene in newborn and infant mice in vivo. Equimolar doses of 1-nitropyrene and 1-nitrosopyrene were used as in the bioassay reported previously. The compounds were administered by ip injection (100 nmol, day 1; 200 nmol, day 8; 400 nmol, day 15). The ethyl acetate soluble metabolites of 1 nitropyrene were identified as 1-aminopyrene, trans-4,5-dihydro-4,5-dihydroxy-1 nitropyrene, 1-nitropyren-3-ol, 1-nitropyren-6-ol, and 1-nitropyren-8-ol on the basis of cochromatography with synthetic standards in two different HPLC systems. Nitroreduction of 1-nitropyrene to 1-aminopyrene was observed only in 1 day old mice. Ethyl acetate soluble metabolites of 1-nitrosopyrene were identified as 1 aminopyrene and 1-nitropyrene. The capacity of 1 day old mice to metabolize 1 nitropyrene and 1-nitrosopyrene exceeded those of 8 and 15 day old mice. The extent of nitroreduction of 1-nitrosopyrene exceeded that of 1-nitropyrene. A major DNA adduct, N-(deoxyguanosin-8-yl)-1-aminopyrene, was identified and quantified in liver and in lung, 24 h after carcinogen treatment. The extents of formation of this adduct (pmol/mg of DNA, mean of two experiments) were as follows: 1-nitropyrene (liver, 4.1; lung, 1.2); 1-nitrosopyrene (liver, 30.4; lung, 6.3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979740 TI - Mammalian synthesis of nitrite, nitrate, nitric oxide, and N-nitrosating agents. PMID- 2979742 TI - The role of nucleophilicity in the ratio of primary to secondary solute-derived products in the decomposition of (1-acetoxypropyl)propylnitrosamine. AB - The reactions of 1-propanediazotic acid, generated in situ from the hydrolysis of (1-acetoxypropyl)propylnitrosamine (1-APPN), with inorganic halides (X- = Cl-,Br ,I-) were studied in pH 8.0 buffer at 37 degrees C with specific emphasis on the extent of structurally rearranged solute-derived product. There is a direct correlation between the nucleophilicity of the halide and increased total and individual isomeric propyl halide (PrX) products. However, the relative formation of rearranged secondary propyl halide (2-PrX) is inversely proportional to solute nucleophilicity. The extent of isomerization ranges from 28% for Cl- to 13% for I . In contrast, 2-propanol accounts for 33% of the total solvent-derived propanol yield. It is also demonstrated that the ratio of 1-PrX/2-PrX is inversely related to [X-] for Br- and I-. The data suggest that a nucleophilicity-driven association of the solute with a propanediazonium ion in the transition state leads to both primary and secondary products. The results are also discussed in terms of understanding the solvolysis products and DNA adducts previously observed from the metabolism of dipropylnitrosamine and the hydrolysis of N propyl-N-nitrosourea. PMID- 2979741 TI - Pyrrolo[1,4]benzodiazepine antitumor antibiotics: relationship of DNA alkylation and sequence specificity to the biological activity of natural and synthetic compounds. AB - The DNA alkylation and sequence specificity of a group of natural and synthetic pyrrolo-[1,4]benzodiazepines [P(1,4)Bs] were evaluated by using an exonuclease III stop assay, and the results were compared with in vitro and in vivo biological potency and antitumor activity. The P(1,4)B antibiotics are potent antitumor agents produced by various Actinomycetes, which are believed to mediate their cytotoxic effects by covalent bonding through N-2 of guanine in the minor groove of DNA. In this article we describe the results of a sensitive DNA alkylation assay using exonuclease III which permits both estimation of the extent of DNA modification as well as location of the precise guanines to which the drugs are covalently bound. Using this assay, we have evaluated a series of natural and synthetic compounds of the P(1,4)B class for their ability to bind to DNA and also determined their DNA sequence preference. The compounds included in this study are P(1,4)Bs carrying different substituents in the aromatic ring, having varying degrees of saturation in the five-membered ring, or differing in the stereochemistry at C-11a. These same compounds were evaluated for in vitro cytotoxic activity against B16 melanoma cells, for potency in vivo in B6D2F1 mice (LD50), and for antitumor activity (ILSmax) against P388 leukemia cells. A good correlation was found between extent of DNA alkylation and in vitro and in vivo potency. Furthermore, on the basis of electronic and steric considerations, it was possible to rationalize why those compounds that showed negligible biological activity were unable to bond covalently to DNA. Last, we have determined that the degree of saturation in the five-membered ring of the P(1,4)Bs has a significant effect on the DNA bonding reactivity and biological activity of this class of compounds. PMID- 2979743 TI - A phospholipase C protocol for phospholipid peroxidation analysis. AB - A new method has been developed to analyze the primary products of phospholipid peroxidation. The procedure utilizes the ability of phospholipase C to hydrolyze phospholipid hydroperoxides to their corresponding diacylglycerol derivatives. 1 Palmitoyl-2-linoleoylphosphatidylcholine (1P,2L-GPC), 1-stearoyl-2 linoleoylphosphatidylcholine (1S,2L-GPC), and 1-stearoyl-2 arachidonylphosphatidylcholine (1S,2A-GPC) were autoxidized. The diacylglycerol hydroxides derived from the phosphatidylcholine hydroperoxides were separated by reverse-phase high-pressure liquid chromatography (RP-HPLC) and normal-phase high pressure liquid chromatography (NP-HPLC). 1P,2L-diglyceride (1P,2L-DG) and 1P,2A DG products were easily separated from 1S,2L-DG and 1S,2A-DG products by RP-HPLC. The linoleate diglyceride oxidation mixture was separated into the 13-trans/cis, 13-trans/trans, 9-trans/cis, and 9-trans/trans isomers by NP-HPLC. Likewise, 1P,2A-DG and 1S,2A-DG oxidation products were resolved into the 15-trans/cis, 15 trans/trans, 12-trans/cis, 11-trans/cis, 9-trans/cis, 8-trans/cis, and 5 trans/cis isomers. In both of the above cases, the 1,2-diacylglycerol isomers could be separated from the 1,3 isomers. Moreover, the diastereomers of the 9-, 8 , and 5-hydroxides could be separated. Each of the diacylglycerol oxidation products was characterized by (1) proton nuclear magnetic resonance (proton NMR), (2) electron ionization-mass spectrometry (EI-MS), and (3) NP-HPLC of the corresponding fatty acids. The diacylglycerol analysis provided the same results for the autoxidation of 1P,2L-GPC as the fatty acid methyl ester analysis. In addition, when 1S,2A-GPC was autoxidized in the presence of 5% alpha-tocopherol, both diastereomers of the 5-hydroxide were observed in the same proportions as the other hydroxides. PMID- 2979744 TI - Intra- and intermolecular reactions of 4,4'-diisocyanatodiphenylmethane with human serum albumin. AB - Diisocyanates are highly reactive industrial chemicals that have been shown to possess toxic activity, including potential for allergic sensitization. To assist in diagnosis of sensitization, immunoassays for diisocyanate-specific antibodies are performed; such assays require preparation of diisocyanate-containing hapten protein conjugates. Conditions were investigated for formation of conjugates yielding varying degrees of hapten binding. Relative concentrations of haptens and proteins were varied as were pH, temperature, and time of reaction. Quantitation of 4,4'-diisocyanatodiphenylmethane (MDI) binding with human serum albumin (HSA) was assessed by absorbance of the isolated conjugates at 250 nm after determination of the molar extinction coefficient for MDI. At pH 7.4 and 37 degrees C, the binding reaction was found to be biphasic with binding of 5-6 mol of MDI groups/mol of HSA within the first minute, followed by incorporation at a rate of 0.16 mol/min during the next 2 h. Evaluation of reaction products using SDS-PAGE revealed extensive inter- and intramolecular cross-linking of HSA by MDI. Intramolecular cross-linking was accompanied by an increased migration of conjugates from an initial molecular mass of 66 kDa, typical of HSA, to a molecular mass of 44 kDa. The change in migration was also produced by using disuccinimidyl tartarate (DST) as hapten and was eliminated when DST was cleaved with sodium periodate. It was attributed to altered protein shape. Conditions that favored binding of MDI with HSA were a high relative concentration of MDI:HSA, a pH of 9.4, and a temperature of 37 degrees C. Under such conditions it was calculated that 53 mol of MDI were bound per mole of HSA after 24 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979745 TI - Diisocyanate antigens that detect specific antibodies in exposed workers and guinea pigs. AB - Evaluation of the immunologic contribution to the pathogenesis of isocyanate lung disease necessitates preparation of isocyanate-protein conjugates to detect anti isocyanate antibodies. The preparation and characterization of the conjugates were described in the preceding paper in this issue. Sera were obtained from two guinea pigs immunized with 4,4'-diisocyanatodiphenylmethane (MDI) and from three workers with occupational exposure to MDI. By use of Western blot analysis, guinea pig IgG antibodies were best detected by the monomeric component of MDI guinea pig serum albumin. ELISA additionally indicated that conjugates which contained a high density of hapten detected greater amounts of antibody than did conjugates containing low amounts of hapten. The same procedures were then used to assess the amount of MDI-specific IgG and IgE antibody in sera from symptomatic workers. Effective MDI-HSA antigens were those that were monomeric and had high haptenic content. Highly substituted conjugates of monoisocyanates (phenyl isocyanate and p-tolyl isocyanate) with serum albumins were also effective in detecting antibodies to MDI. These results indicate the importance of the composition of isocyanate conjugate antigens in detecting antibodies to diisocyanates and suggest that standards be developed for preparation and characterization of these diagnostic serologic reagents. PMID- 2979746 TI - Stereochemistry of the major rodent liver microsomal metabolites of thecarcinogen dibenz[a,j]acridine. AB - The major metabolites of the carcinogen dibenz[a,j]acridine formed in rodent liver microsomal preparations were trans-3,4-dihydroxy-3,4 dihydrodibenz[a,j]acridine (DBAJAC-3,4-DHD) and dibenz[a,j]acridine 5,6-oxide (DBAJAC 5,6-oxide) [Gill et al. (1987) Carcinogenesis 8, 425-431]. The enantiomers of DBAJAC-3,4-DHD were prepared from the separable diastereoisomeric esters with (+)-endo-1,4,5,6,7,7-hexachlorobicyclo[2.2.1]hept-5-ene-2-carboxyl ic acid (HCA). The absolute configuration of trans-3(R),4(R)-dihydroxy-1,2,3,4 tetrahydrodibenz[a,j]acridine was assigned by conversion to the bis[p (dimethylamino)benzoate] and examination of the exciton coupling in its circular dichroic (CD) spectrum. The 3(R),4(R)-tetrahydrodiol was converted to DBAJAC 3(R),4(R)-DHD. The enantiomers of DBAJAC 5,6-oxide were partially resolved by chiral stationary-phase chromatography, and subsequent methoxide attack afforded two enantiomerically enriched isomeric ethers from each fraction. The structures of the two ethers from each enantiomer were determined, and from their 1H NMR spin-spin coupling between the H5 and H6 signals and the CD spectra of the ethers, the absolute configuration of the ethers, and hence the 5,6-oxides, was determined. The enantiomeric composition of the 3,4-dihydrodiol and 5,6-oxide formed as microsomal metabolites of rat liver preparations was 69% 3R,4R and 81% 5R,6S, respectively. When rats were pretreated with 3-methylcholanthrene (MC), these percentages were 70% and 5%, indicating a reversed stereochemical preference for oxide formation in the MC-induced preparation. Results are also presented for phenobarbitone-induced rat liver and mouse liver preparations. PMID- 2979747 TI - Metabolism of profluralin in rats. 1. Identification of metabolites. AB - Fluorine NMR spectroscopy has been used to identify and quantitate metabolites of profluralin appearing in the urine of rats postingestion of the herbicide. The chemical shift of the trifluoromethyl group was shown to be a useful indicator of the chemical type of metabolite, and the shifts for the classes of materials previously identified as metabolites have been determined. By use of chemical shift criteria this work provides evidence for several metabolites that have not been recognized in previous work; some of these remain incompletely characterized. Though the number and types of metabolites detected in the urine were consistent from animal to animal, the time course for appearance of metabolites was found to be animal dependent. PMID- 2979748 TI - Sequence selectivity of DNA covalent modification. PMID- 2979749 TI - Diazonium ion derived products from the Ce(IV) oxidation of beta-hydroxy nitrosamines. PMID- 2979751 TI - Enantioselective synthesis and preliminary metabolic studies of the optical isomers of 2-n-propyl-4-pentenoic acid, a hepatotoxic metabolite of valproic acid. AB - The enantiomers of 2-n-propyl-4-pentenoic acid (delta 4-VPA), a known hepatotoxic metabolite of 2-n-propylpentanoic acid (valproix acid, VPA), were synthesized with the aid of the chiral auxiliaries (4S)-4-(2-propyl)-2-oxazolidone and (4R,5S)-4-methyl-5-phenyloxazolidone. Alkylation of the n-valeryl derivatives of these oxazolidones with allyl bromide, followed by reductive cleavage and chromic acid oxidation of the product, afforded the desired acids, (R)- and (S)-delta 4 VPA. Greater than 93% enantiomeric excess was achieved in the preparation of both enantiomers. Preliminary studies on the metabolic fate of (R)- and (S)-delta 4 VPA in freshly isolated rat hepatocytes revealed striking differences in the biotransformation of the two enantiomers. Quantification of two major metabolites of delta 4-VPA, viz., 4,5-diOH-VPA gamma-lactone and 2-n-propyl-2(E),4 pentadienoic acid (delta 2E,4-VPA), indicated that larger amounts of the gamma lactone were formed in incubations utilizing (R)-delta 4-VPA as substrate, whereas production of the diene was greater in incubations with (S)-delta 4-VPA. On the basis of the premise that delta 4-VPA serves as a mechanism-based irreversible inhibitor of enzymes of the fatty acid beta-oxidation complex, these differences in metabolism suggest that the two enantiomers of delta 4-VPA may differ in their hepatotoxic potential. PMID- 2979750 TI - Biochemical characterization of two hemorrhagic proteases from the venom of Lachesis muta (bushmaster). AB - Two hemorrhagic proteases, Lachesis hemorrhagic toxins a and b (LHTa and LHTb), were isolated from the venom of Lachesis muta, which is distributed in Central and South America. One protease showed strong hemorrhagic action, while the other showed weak hemorrhagic activity even though the two enzymes are very similar in their chemical properties. Neither enzyme hydrolyzed arginine esters, but both hydrolyzed casein and reduced fibrinogen. The A alpha chain of fibrinogen was hydrolyzed first, and the B beta chain was hydrolyzed later. The gamma chain of fibrinogen was resistant to hydrolysis. The molecular weights of LHTa and LHTb were very similar, 22,000 and 23,000, respectively. The amino acid composition of LHTa was also similar to that of LHTb. The secondary structure of LHTa as determined by Lippert's equation was 52% alpha helix, 17% beta sheet, and 31% random coil; that of LHTb was 47% alpha helix, 13% beta sheet, and 40% random coil. PMID- 2979752 TI - Synthesis and tumor-initiating activities of dimethylchrysenes. AB - Previous studies have shown that 5-methylchrysene (5-MeC) is more carcinogenic on mouse skin than the other methylchrysenes and that the structural requirements favoring tumorigenicity of methylated polynuclear aromatic hydrocarbons are the presence of a bay region methyl group and free peri position, both adjacent to an unsubstituted angular ring. The purpose of this study was to extend these structure-activity relationships to dimethylchrysenes. The following dimethylchrysenes were synthesized: 1,5-dimethylchrysene (1,5-diMeC), 5,6-diMeC, 5,7-diMeC, 5,12-diMeC, 1,6-diMeC, 6,7-diMeC, and 6,12-diMeC. Bioassays of these compounds for tumor-initiating activity on mouse skin demonstrated that all were significantly less tumorigenic than 5-MeC; only 5,6-diMeC had significant tumorigenic activity. Since the relatively low activities of 5,7-diMeC and 5,6 diMeC were unexpected on the basis of the structural requirements stated above, anti-1,2-dihydroxy-3,4-epoxy-1,2,3,4-tetrahydro-5,7-dimethylchrysene+ ++ (anti 5,7-diMeC-1,2-diol-3,4-epoxide) was synthesized. Its mutagenicity in Salmonella typhimurium and reactivity with calf thymus DNA were compared to those of the major ultimate carcinogen of 5-MeC, anti-5-MeC-1,2-diol-3,4-epoxide. It was strongly mutagenic (2500 revertants/nmol), although less active than anti-5-MeC 1,2-diol-3,4-epoxide (7200 revertants/nmol). Its reactivity with calf thymus DNA was similar to that of anti-5-MeC-1,2-diol-3,4-epoxide. The results of this study demonstrate that the structural requirements which favor tumorigenicity of monomethylchrysenes are not sufficient for high tumorigenicity of dimethylchrysenes. PMID- 2979753 TI - Styrene oxide as a stereochemical probe for the mechanism of aralkylation at different sites on guanosine. AB - The stereochemistry at the alpha-carbon atom of the styrene moiety in styrene oxide-guanosine products was established, and the stereochemical consequences of reaction of optically active styrene oxide with guanosine were investigated. Inversion of stereochemistry in products at the alpha-carbon of styrene oxide decreased in the sequence 7- much greater than N2- greater than O6-substituted guanosines. These findings show that an extensively ionized substrate is needed for reaction at the exocyclic N2 and O6 sites on guanosine but that the reactive intermediate is not an ideal planar trigonal carbonium ion. This follows because inversion exceeds retention in both of the exocyclic substituted products. PMID- 2979754 TI - A chemical comparison of methanesulfonyl fluoride with organofluorophosphorus ester anticholinesterase compounds. AB - The chemical reactivity and physical properties of methanesulfonyl fluoride (MSF) were quantitatively measured for direct comparison with those of isopropyl methylphosphonofluoridate (GB, sarin) and diisopropyl phosphorofluoridate (DFP). The chemistry involved reaction rate constant measurements of MSF hydrolysis and for reactions with phenolic, amine, oxime, hydroxamic acid, phenyl N hydroxycarbamate, and hydroxylamine compounds and cupric imidazole and bipyridyl complexes. Analogous chemistry with GB and DFP occurred only for the nucleophilic reactions. MSF was substantially less reactive toward interactions involving hydrogen bonding, cupric complex coordination, and quaternary amine formation. Physical characterization included water solubility, vapor pressure, heat of vaporization, solubility parameter, hydrogen bond basicity, UV, IR, and NMR measurements. MSF is a much weaker hydrogen bond acceptor than the phosphorus esters. The hydrogen bond basicity beta-scale measurement provides a key insight to the chemical reactivity divergence of MSF and DFP or GB. Its unexpectedly low value is interpreted that MSF does not possess a site with sufficient basicity to be protonated by strong acid, to enter into hydrogen bonding, or for coordination with the cupric ion. PMID- 2979755 TI - Fate of free radicals generated during one-electron reductions of 4-alkyl-1,4 peroxyquinols by cytochrome P-450. AB - Free radicals resulting from the one-electron reduction and subsequent homolytic cleavage of oxygen-oxygen bonds by heme proteins are likely to be responsible for some aspects of the toxicity of organic hydroperoxides. In the present work, effects of the 4-alkyl substituent of 2,6-di-tert-butyl-4-alkyl-4 hydroperoxycytohexa-2,5-dienones (1,4-peroxyquinols) on radical production were investigated with microsomal cytochrome P-450 from rat liver. Quinoxy radicals from homolysis of the peroxyquinols underwent beta-scission to produce a quinone and an alkyl radical, and this process occurred with increasing frequency as the stability of the alkyl radical increased. The fate of benzyl and 2-phenylethyl radicals generated from the appropriately substituted peroxyquinols was investigated also. The former was converted to benzyl alcohol, benzaldehyde, and toluene and the latter to 2-phenylethanol, phenylacetaldehyde, ethylbenzene, styrene, and benzaldehyde. Oxygen-18 labeling studies demonstrated that 80-85% of the benzyl alcohol incorporated oxygen from the hydroperoxide and the balance from molecular oxygen. This indicates that the predominant reaction pathway involved recombination between the benzyl radical and the iron-bound hydroxyl radical of the P-450 intermediate complex. By contrast, about 50% of 2 phenylethanol from the 2-phenylethyl radical incorporated oxygen from water and the balance from O2. Two alternative mechanisms are proposed to explain the formation of 2-phenylethanol that contained oxygen from water and the concurrent formation of styrene: (a) oxygen exchange of the P-450 intermediate with water, followed by hydrogen abstraction and radical recombination reactions with the P 450 complex, or (b) oxidation of the radical to the 2-phenylethyl cation followed by proton elimination and hydration. PMID- 2979756 TI - Synthesis and properties of H-ras DNA sequences containing O6-substituted 2' deoxyguanosine residues at the first, second, or both positions of codon 12. AB - Nine 16-base oligodeoxyribonucleotides having the sequence of codons 9 through the first base of codon 14 of the rodent H-ras gene, i.e., 5' d(GTGGGCGCTG*G*AGGCG)-3', have been synthesized containing either an O6-methyl- (G* = m6G), O6-ethyl- (G* = e6G), or the newly described O6-benzyl-2' deoxyguanosine residue (G* = b6G) at position 10 and/or 11 from the 5'-end. The conversion of the protected O6-substituted 2'-deoxyguanosine derivatives to the corresponding 3'-[O-(2-cyanoethyl) diisopropylphosphoramidites] and their incorporation into oligodeoxyribonucleotides were conveniently accomplished by using an "in situ" activation approach and automated phosphite triester synthetic methods. These oligomers were characterized by enzymatic digestion to their component nucleosides and were shown to be free of detectable contamination by known nucleoside impurities that can be produced during these syntheses. The melting behavior and circular dichroism spectra are described for duplexes of the nine O6-substituted 2'-deoxyguanosine containing oligomers paired with the complementary strand 5'-d(CGCCTCCAGCGCCCAC)-3', and these data have been compared with those for the "wild-type" unsubstituted duplex. PMID- 2979757 TI - Kinetic studies of the reactions of benzo[a]pyrene-7,8-diol 9,10-epoxides in aqueous solutions of human serum albumin and nonionic micelles. AB - The rates of reactions of the cis and trans (benzylic 7-hydroxyl group relative to epoxide oxygen) bay-region 7,8-diol 9,10-epoxides of benzo[a]pyrene (DE-1 and DE-2, respectively) in solutions containing human serum albumin (HSA) and a model system of micelles of the nonionic detergent Tween-80 have been determined as a function both of HSA or Tween-80 concentrations and of pH. The effect of increasing concentrations of both HSA and Tween-80 is to retard substantially the rates of reaction of DE-1 and DE-2 over the pH range 5-7. The rate data are consistent with a mechanism in which the diol epoxides physically associate with HSA or Tween-80, and the rates of reaction of the association complexes are reduced compared to those of free diol epoxides. The limiting rate constants for reaction of the diol epoxide.HSA and diol epoxide.Tween-80 complexes are dependent on pH. The rate data are best accommodated by a mechanism in which the complexes react by two competing pathways, one whose rate is proportional to hydronium ion activity, and the second whose rate is pH-independent. These reactions of the association complexes are therefore kinetically analogous to the acid-catalyzed and spontaneous reactions of free diol epoxide. The spontaneous reaction of the (DE-2).HSA complex results in approximately 10% covalent binding of diol epoxide to the protein, whereas the acid-catalyzed reaction of the complex results in significantly less covalent binding. The balance of the products consists of tetraols formed by hydrolysis of the diol epoxide. PMID- 2979758 TI - Human tumor xenografts in the nude mouse and their value as test models in anticancer drug development (review). AB - In an attempt to increase the predictability of preclinical antitumor testing, the value of human tumor lines in immune-deficient nude mice is assessed by reviewing the relevant literature. This test model is rather elaborate due to the nature of the animal as well as test and evaluation procedure. However, it represents a realistic simulation of clinical drug treatment. This is demonstrated by (a) a good correlation of drug effects in the nude mouse with clinical results in the donating patient's tumor and (b) by a good predictability of a panel of human tumor lines for clinically effective drugs. In order to avoid clinical trials with inactive drugs and no therapeutic benefit for a large number of patients, the application of human tumor xenografts in anticancer drug development is warranted. PMID- 2979759 TI - Clastogenic action of chemotherapy in humans (review). AB - Examinations of the chromosome damaging (clastogenic) action of chemotherapy not only present valuable insights into the genetic hazards for man resulting from the respective drugs, but also can be used for control of success of certain treatments. In addition, those studies may be helpful in detecting "high-risk" individuals and, in consequence, in protecting them from the detrimental action of therapeutics they are highly sensitive to. Clastogenic activity should not be discussed without considering the close correlation between induction of mutations (e.g. chromosome changes) and malignant transformation, as documented by recent evidence on the meaning of point mutations as well as of chromosomal rearrangements in oncogene activation. PMID- 2979760 TI - Inhibitory effects of some organometallic complexes of rhodium(I) and iridium(I) on carrageenin paw edema in rats. AB - A group of 4 organometallic complexes of Rhodium (I) and 1 Iridium(I) was tested for the evaluation of their anti-inflammatory activity on carrageenin paw edema in rats. All the compounds used inhibited the development of paw edema by more than 50% at different dose-levels. The activity of the pyridinalmethylimine derivative [Rh(COD)PMI]+ Cl- had better results than that of [Rh(NBD)PMI]+ Cl- and even more than the dimeric complexes tested. The higher activity of [Ir(COD)Cl]2 as compared with [Rh(COD)Cl]2 suggests that it would be of interest to examine further Iridium(I) complexes, among which [Ir(COD)PMI]+ Cl- could be a good candidate. PMID- 2979761 TI - The induction of esophageal tumors in mice: dose and time dependency. AB - Three hundred and forty eight C57Bl mice were killed after diethyl-nitrosamine (DEN) treatment at various time intervals ranging from one day to six months. No esophageal tumors occurred in the first three months after DEN treatment; They occurred, however, at four months and increased in number at six months of DEN treatment. Other groups of animals under the same initial DEN treatment were allowed to survive seven or nine months without treatment. One esophageal tumor was recorded at observations made seven months after one single day of DEN administration. A significant increase in the number of esophageal tumors occurred at seven months in mice treated with DEN for two weeks, and for one, two, three or four months; the highest tumor frequency was found in mice treated for six months and surviving three additional months on a carcinogen-free diet. These results suggest that clones of esophageal cells had been "programmed" for tumor growth at an early stage of DEN treatment. The tumors had, however, remained undetected at macroscopical and microscopical examination several months previously. It is apparent that not only the dose administered, but also the post carcinogen interval is an important factor in esophageal carcinogenesis in the mouse. PMID- 2979763 TI - Animal experiments as a model for clinical trials of a new dosage form of mitomycin C for peritoneal carcinomatosis. AB - Rabbits with peritoneal carcinomatosis induced by VX2 carcinoma were used as an experimental model to study a new dosage form of mitomycin C, MMC-CH, comprising activated carbon adsorbing mitomycin C. Rabbits revived an intraperitoneal injection of mitomycin C (1 mg/kg in the form of MMC-CH or 0.185 mg/kg in the form of mitomycin C solution). The two doses were equal in toxicity. MMC-CH extended survival time measured in days to 139% as compared to mitomycin C solution. Autopsy findings showed peritoneal carcinomatosis to be much less developed with MMC-CH than with mitomycin C solution. Hematological and blood biochemical analyses showed no remarkable abnormality. PMID- 2979762 TI - Cancer induction by the sulfate form of 4-(hydroxymethyl) benzenediazonium ion of Agaricus bisporus. AB - The sulfate form of 4-(hydroxymethyl) benzenediazonium ion (HMBD) of Agaricus bisporus was given by 26 weekly subcutaneous injections at 50 micrograms/g body weight to Swiss mice. As a result of treatment, tumors of subcutis and skin developed in 32% and 14% of the females and 40% and 4% of the males, respectively. The corresponding tumor incidences in the sodium sulfate-injected solvent control groups were 0% and 2% in females and 4% and 0% in males, respectively. Histopathologically, the tumors were fibrosarcomas, rhabdomyosarcomas and angiosarcomas of the subcutis, squamous cell papillomas and carcinomas of skin. The present finding discounts an earlier allegation that the tetrafluoroborate portion of the diazonium ion acted as a cocarcinogen. HMBD was found at a 0.6 ppm level in the mushroom of commerce Agaricus bisporus. PMID- 2979764 TI - In vivo characterization of P388 leukemia resistant to mitomycin C. AB - A line of P388 leukemia resistant to mitomycin C (MMC) was successfully developed in vivo by treating mice bearing parental P388 (P388/0) with MMC followed by serial passage of the surviving leukemic cells. From this P388/MMC line, a subline was derived by not treating the passage mice with MMC (P388/MMC-NP); resistance to MMC was stable for as many as 56 weeks of transplantation. The chemosensitivities of each P388 line to assorted anticancer drugs were compared in vivo. Both P388/MMC and P388/MMC-NP had similar patterns of drug cross resistance and collateral sensitivity. With respect to alkylating agents (e.g. cyclophosphamide, Platinol and chlorambucil), there was generally a partial degree of cross-resistance, sometimes only detectable at suboptimal dose levels. With respect to DNA binders or intercalators (e.g. actinomycin D, luzopeptin A, amsacrine, doxorubicin), the extent of cross-resistance varied from none (dihydoxyanthraquinone) to marked (doxorubicin). Antimitotic inhibitors (vinblastine and vincristine) were completely cross-resistant, as were some miscellaneous natural agents (rebeccamycin, VP-16, sesbanimide, and elsamicin, a chartreusin analog). Antimetabolites (e.g. methotrexate and 6-thioguanine) showed no cross-resistance and even demonstrated some occasional evidence of collateral effectiveness. PMID- 2979765 TI - Non-gradient regional differences in tumor growth. AB - Tumor cells inoculated subcutaneously into various regions of the trunk of dogs showed differences in growth for prolonged time periods. However, consistent antero-posterior gradient tumor growth patterns previously reported by other investigators in mouse tumor systems were not observed. The regional variance in tumor growth was not great enough to overcome the dose-response growth relationship in the dogs inoculated with doubling doses of tumor cells. PMID- 2979766 TI - First--generation human renal cell carcinoma xenografts in the Nb rat subrenal capsular assay model. AB - This laboratory has evaluated the subrenal capsular assay system previously in the Nb rat with the Nb rat prostate adenocarcinoma model. Human renal cell carcinoma xenografts are now being employed in our Nb rat SRC model, the advantage of this system being that one can determine the effect of chemotherapy in a very short period of time, i.e., 7 days. This method saves considerable time and expense and may serve as a useful indicator for effective chemotherapeutic agents in individual cancer patients. In our studies, cyclophosphamide was the only agent to produce a significant effect on tumor volume. PMID- 2979767 TI - Lymphoid tissue-associated colonic adenocarcinomas in rats. AB - Eight months after a single s.c. injection of 1,2-dimethylhydrazine (21 mg/kg b.w.) to male Sprague-Dawley rats, seven colonic tumors (i.e. 17.5%) were found at autopsy. Histological examination revealed invasive adenocarcinomas in all seven tumors. All tumors were seen to originate in pre-existing discrete lymphoid aggregates. Adenomas or adenocarcinomas arising from pre-existing adenomas were not found. Previous experiments, using the same strain of rats, demonstrated the occurrence of adenomas with or without invasive adenocarcinomas after multiple doses of DMH. The causes responsible for the dose-dependent specificity in the histogenesis of colonic tumors in our model remains to be elucidated. PMID- 2979768 TI - Autografting colonic adenocarcinomas into the colonic submucosa in rats. AB - After inducing colonic tumors in 47 rats by weekly subcutaneous injections of 1,2 dimethylhydrazine, all rats were laparotomized and tumors were autotransplanted to tumor-free areas of the transverse colon. True autotransplants were considered to be tumors occurring in the grafted areas, which were covered by normal colonic mucosa. The frequency of auto-grafted tumors was 35.5%. In a previous work, a low incidence of 1,2-dimethylhydrazine-induced tumors was found in the transverse colon of the rat. The present results suggest that the failure of tumor growth in the transverse colon may be unrelated to an intrinsic natural "resistance" of that segment of the colon to tumor growth. PMID- 2979769 TI - Effects of aminothiols in 2-acetylaminofluorene-treated rats. III. Metabolic activation of aromatic amines. AB - Six groups of Wistar rats received a standard diet supplemented with 0.05% 2 acetylaminofluorene and/or 0.1% natural (reduced glutathione) or synthetic (N acetyl-L-cysteine) aminothiols. The discontinuous feeding regimen consisted of 4 cycles, each composed of 3 weeks of treatment followed by withdrawal for 1 week. At the 3rd and 4th week of each cycle, the liver was removed from 4-5 rats within each group, and pools of S-12 fractions were assayed for the ability to activate 2-acetylaminofluorene and other aromatic amines, either structurally related (i.e. 4-acetylaminofluorene and 2-aminofluorene) or unrelated (i.e. 2 naphtylamine and benzidine) to mutagenic metabolites in strain TA98 of S. typhimurium. In untreated rats, there was a consistent and marked trend to an age dependent loss of metabolic activation of all test compounds during the 16 weeks of the experiment. Feeding of 2-acetylaminofluorene resulted in an evident autoinduction of metabolism which was continuously amplified with time, even during the withdrawal weeks. In the same animals, activation of the other amines was initially inhibited but then progressively shifted to a mild cross-induction which, in the case of the structurally related compounds, became significant at the end of the 4-cycle treatment with 2-acetylaminofluorene. The metabolic effects of the two aminothiols were broadly variable, depending on the thiol, on its co-administration with 2AAF, on the week and cycle of treatment, and on promutagens tested. PMID- 2979770 TI - Differences in DNA content and proliferation rate between adenocarcinomas of the uterine cervix and corpus. AB - The DNA content and the S-phase rate of 26 cervical adenocarcinomas and 100 endometrial adenocarcinomas, determined by flow cytometry, were compared. Statistically significant differences in DNA patterns were found, although only pure adenocarcinomas were studied in both groups. Peri-diploid values (1.8c-2.2c) were less common in cervical adenocarcinomas (38%) than in endometrial carcinomas (65%) (p less than 0.05). Six (23%) of the cervical carcinomas and 8 (8%) of the endometrial carcinomas had two detectable stem lines. The mean S-phase rate was significantly higher in adenocarcinomas of the uterine cervix than in endometrial carcinomas (p less than 0.02). The dominance of aneuploid values, the high proportion of polyclonal tumors and the high S-phase rates of the cervical adenocarcinomas have more similarities with the DNA patterns of cervical squamous cell carcinomas than with those of endometrial adenocarcinomas. PMID- 2979771 TI - Diurnal variation in the therapeutic efficacy of 5-fluorouracil against murine colon cancer. AB - Mice bearing the murine colon carcinomas Colon 38 or Colon 26 were treated with 5 fluorouracil (5 FU) (60 or 100 mg/kg, respectively) weekly for 4 weeks at 08.30 hr or at 18.30 hr. The antitumor effect of treatment at 08.30 hr was significantly better for both types of tumors, but most pronounced for Colon 38. Toxicity was evaluated after administration of 60 mg 5 FU/kg. No thrombocytopenia was observed. Leucopenia was observed after treatment at 18.30 hr. It is concluded that the therapeutic efficacy of 5 FU against murine colon cancer is higher when 5 FU is administered at 08.30 hr. PMID- 2979772 TI - Study of oncogenic potentialities of human melanoma: identification of N-ras oncogene after DNA transfer and tumour induction. AB - The oncogenic potentialities of human melanoma cells derived from two different patients were studied using DNA-mediated gene transfer into NIH 3T3 cells followed by tumor induction into athymic nude nice. 64% of the mice injected subcutaneously with selected cells which had been co-transfected with human melanoma DNA and the selective marker NeoR developed tumors within 3-4 weeks, while up to 100% of those injected with cells transfected three days before with melanoma DNA developed tumors within 4-6 weeks. Southern blots analysis of the tumors indicated that almost all of them contained human sequences. Hybridization with different oncogene probes showed the presence of an human Eco RI N-ras hybridizing fragment in the primary and secondary derived tumors, indicating that a transforming N-ras oncogene in human melanoma had been transferred to recipient cells and that transformed cells induced tumors in nude mice. PMID- 2979773 TI - Development and applications of an animal model for human tumor xenografts. AB - Wistar rats treated with cyclophosphamide (4x 10 mg/kg), total lymphoid irradiation (9.0 Gy; dose rate 0.60 Gy/min) and cyclosporin A (15 mg/kg, daily, orally) developed a state of immune suppression permitting the growth of human tumor xenografts. Immunosuppression was monitored by lymphocyte counts, serum IgG determination. PHA and Con A lymphocyte-responses, proportion of B cells and histopathological studies of the lymphoid organs. The lymphocyte counts, IgG levels, PHA and Con A stimulation values remained severely depressed, during the period of cyclosporin A administration. Repopulation of the paracortical areas of the lymph nodes and the peri-arteriolar sheaths of the spleen did not occur, neither the reconstruction of the germinal centers in these organs. The thymus underwent severe atrophy. Seven of eight different types of human tumors were successfully xenografted in the immunomodified rat. The xenografted tumors maintained their original morphologic features and the mitotic rate did not change during subsequent transplantations. PMID- 2979775 TI - Effects of aminothiols in 2-acetylaminofluorene-treated rats. II. Glutathione cycle and liver cytosolic activities. AB - Reduced glutathione, enzymes involved in its metabolism and other cytosolic activities were evaluated in liver preparations of Wistar rats fed with a diet supplemented with 2-acetylaminofluorene (0.05%) and/or with glutathione or N acetyl-L-cysteine (0.1%). The treatment lasted 4 cycles, each composed of 3 weeks of special diet followed by 1 week of standard diet. The carcinogen produced a considerable increase in gamma-glutamyl transpeptidase in liver homogenates at cycles III and IV, with an irreversible trend which was not discontinued even during the weeks of standard diet. Moreover, generally from cycle I, 2 acetylaminofluorene stimulated several enzyme activities in the liver cytosol, such as glutathione S-transferase, glutathione reductase, glucose 6-phosphate dehydrogenase, NADH- and NADPH-dependent diaphorases. Administration of the two aminothiols to untreated rats resulted in a significant enhancement of glutathione peroxidase, glucose 6-phosphate dehydrogenase and diaphorases. In 2 acetylaminofluorene-treated rats, both thiols further stimulated glutathione S transferase during the last treatment cycles and attenuated gamma-glutamyl transpeptidase activity, which however was not sufficient to thoroughly counteract the liver lesions due to the massive feeding of the carcinogen. Hepatocellular glutathione was enhanced during the last cycle of treatment with 2 acetylaminofluorene, and was further increased by co-administration of exogenous glutathione. PMID- 2979776 TI - Trace element changes in serum and skeletal muscle compared to tumour tissue in sarcoma-bearing rats. AB - Cancer cachexia is characterized by wasting of the lean tissue and profound changes in the body composition of the tumour host. These changes are partly explained by an inefficient energy production but other factors may also be important, such as deficiency of essential nutritional components. In the present study the changes of trace elements in serum and skeletal muscle were compared to those in tumour tissue during tumour progression in sarcoma--bearing rats. Trace element analysis was performed directly on serum specimens and frozen sections from skeletal muscle and tumour tissue. The samples were analysed by energy dispersive X-ray fluorescence spectrometry (EDXRF) without any other pre treatment such as homogenization and extraction. In skeletal muscle an increased content of zinc was found during tumour progression. The iron concentration was unchanged, but since muscle wasting is part of the cachexia this means that iron was transferred to other compartments. Thus the iron content of serum was doubled and tumour tissue had a high concentration of iron. Selenium was below detection limits in skeletal muscle but well detectable in tumour tissue and it increased during tumour growth. Rubidium and potassium content correlated in all tissues (R:0.98) as did bromine and sodium (R:0.98). Copper behaved differently from the other trace elements and showed large variability. This was also true when tissue copper was individually correlated to all other trace elements in the same tissue. PMID- 2979774 TI - Vertebrate lectins and their possible role in fertilization, development and tumor biology (review). AB - Agglutination of cells is not only a functional test, employed in lectin characterization: the ability of lectins to mediate intercellular contacts may have physiological relevance beyond the test tube. Selected examples of advances in research on vertebrate lectins underscore their potential physiological significance in fertilization, development and in the pathogenesis of cancer. PMID- 2979777 TI - Effects of 2-(hydroxyphenyl) indole on mammary gland growth and uterine adenomyosis in four strains of mice. AB - Subcutaneous implantation of a silastic tube containing the compound 4b, one of 2 (hydroxyphenyl) indoles, enhanced, while treatment with the compound diluted with cholesterol at the concentration of 1/40 inhibited the formation of preneoplastic mammary hyperplastic alveolar nodules in four strains of mice (SHN/Mei, SLN/Mei, GR/AMei and C3H/HeMei). However, there was an apparent strain-difference in mammary susceptibility to 4b. Furthermore, the development of uterine adenomyosis in response to 4b varied from strain to strain. These results suggest that more attention should be paid to the choice of specific mouse strains for the study of related fields. PMID- 2979778 TI - Effects of 2-allophanyl-2-allyl-4-valerolactone (valofan) on cerebral energy metabolism of rat in normoxia and hypobaric hypoxia. AB - The effects of an atypical barbiturate, valofan, and of a classical barbiturate, exobarbital, on the cerebral energy metabolism of rat have been evaluated under normoxic and hypobaric hypoxia conditions. In hypobaric hypoxia, the survival time was significantly increased by valofan 1.5 g/kg and exobarbital 0.05 g/kg in respect to the controls. In normoxia, valofan 1.5 g/kg (dose weakly sedative) significantly increased creatin-P level and reduced pyruvate and lactate concentrations; exobarbital 0.05 g/kg (anesthetic dose) showed a similar behavior except for glucose which was increased. In hypobaric hypoxia, valofan and exobarbital did not cause any change in the pool of labile phosphates, nor in the levels of pyruvate and lactate in respect to the controls. Repeated treatment (20 days) with the two drugs significantly altered the cerebral energy metabolism during normoxia conditions. The concentrations of ATP, creatin-P, glucose, pyruvate and lactate were increased, while L/P ratio was unaffected. PMID- 2979779 TI - Effect of PSK on the serum level of immunosuppressive substance in splenectomized rats. AB - Tumor-bearing rats with and without splenectomy were used for an immunological study on the effects of PSK, an immunomodulator, with respect to the survival rate and serum level of immunosuppressive (IS) substance. There was no significant difference in survival rates between the tumor-resected and tumor resected-plus-splenectomy groups. The IS substance level in the former group was higher than that in the latter group. Normalization of IS substance level and prolongation of survival were observed in the group administered PSK. PSK can be effective in combination with surgical treatment. PMID- 2979780 TI - Establishment of pharmacological doses of estrogen-dependent mammary tumors with estrogen and progesterone receptors. AB - A new mammary carcinoma (TF15) which was derived from ovary-dependent mammary carcinoma (TF4) and which was dependent on pharmacological doses of estrogen was established. TF15 grew well in ovariectomized rats given injections of 1mg 17 beta-estradiol but not in intact or ovariectomized rats with injections of 0,1, 10 or 100 micrograms 17 beta-estradiol. TF15 was found to be a secretory carcinoma and contained estrogen and progesterone receptors. In general, pharmacological doses of estrogen inhibit the growth of mammary carcinomas with estrogen receptors (ER) and progesterone receptors (PgR) in humans and animals whether they are ovary-dependent or not (1-10). Recently, we obtained mammary carcinomas which are dependent on pharmacological doses of estrogen by subcutaneous implantation of explants from ovary-dependent mammary carcinomas (TF4) in female rats after ovariectomy and injections of low doses of estrogen. In the present paper, the biological, biochemical and morphological characteristics of these carcinomas are reported. PMID- 2979781 TI - Current concept of halothane hepatitis (review). AB - Clinically, halothane is still a useful volatile anesthetic, but since many cases of liver disorders considered attributable to halothane have been reported up to date, a number of studies have been made on the etiology and mechanism of halothane hepatitis. There are at least two possible mechanisms of halothane hepatitis; the first is the direct toxic reaction associated with free radical that is related to reductive pathway enhanced by hypoxia, and the other is the immune--mediated reaction in which the antigen is associated with the oxidative and/or reductive route. However, the etiology and mechanism of halothane hepatitis have yet to be elucidated, and clinically there is no obvious evidence that halothane can induce hepatic disorders. We have concluded at present that the use of halothane should be avoided in patients with liver disorders, in patients under long-term administration of drugs that may induce enzymes involved in halothane metabolism, in patients with high allergic sensitivity, and in patients undergoing operations in which liver circulation is reduced. PMID- 2979782 TI - Dydrogesterone has no peripheral (anti)-androgenic properties. AB - Dydrogesterone, an orally effective progestational compound, frequently used in a large spectrum of endocrine and gynecologic problems was evaluated for possible peripheric androgenic or anti-androgenic activity. The drug was tested in vivo in the hamster flank organ test and in vitro in tests based on molecular mechanisms of androgen action. Dydrogesterone appeared to have neither androgenic nor anti androgenic properties. This is important in view of the fact that dydrogesterone can also be indicated during pregnancy. PMID- 2979783 TI - Effect of estradiol -17- beta (E2) on cell proliferation in the uterus and the MXT mammary tumor borne by intact, ovariectomized and/or hypophysectomized mice. AB - The aim of the present work was to study the effects of estradiol (E2) on cell proliferation in well-differentiated mammary MXT tumors borne either by intact (INT), or by castrated (OVX), or by hypophysectomized (HX), or by castrated and hypophysectomized mice) OV-X-HX). Ovariectomy and hypophysectomy were performed, respectively, 6 days and 5 days before the sacrifice of animals bearing tumors during a 28 day period. Cell proliferation was measured by using an in vivo nuclear labeling with tritiated thymidine (3H-TdR) followed by autoradiography. Uteri were used as controls for the methodology. Our data demonstrate that E2 exerts a significant and transient stimulatory effect on cell proliferation in the uterine luminal and glandular epithelia of OVX or OVX-HX adult mice. This mitogenic enhancement is significantly higher in OVX-HX mice than in the other experimental groups. In MXT tumors, a significant increase of cell proliferation is induced by E2 in INT or OVX mice; however, no enhancement occurs after HX performed either in intact or in castrated mice. In the latter situation (HX), a dramatic increase of basal cell proliferation rates is consistently observed in tumors, as compared to those levels observed in INT or in OVX animals. The paradoxical increase in basal proliferation rates of MXT mammary tumors in HX animals was not suppressed by administration of ovine prolactin. This suggests that other hypothalamo-hypophysial factors might be endowed with direct inhibitory influence on growth of the MXT tumors. PMID- 2979784 TI - The effect of cortisol or ACTH on plasma concentrations of free fatty acids. AB - The effect of infusion of cortisol (14 mumol) or ACTH (Synacthen, 250 micrograms) injection on plasma levels of free fatty acids has been examined in normal men. The increment in plasma cortisol levels after infusion of cortisol (+182%) was much less than after ACTH injection (up to 262%). Mean plasma levels of free fatty acids were only significantly increased after ACTH injection. However, some inter-subject variation occurred and a small but significant increment in plasma levels of free fatty acids was seen for one control subject and for a subject who received an infusion of cortisol. It is possible that the reported diurnal variations in plasma levels of free fatty acids may be related to changes in plasma levels of ACTH or cortisol throughout a 24h period. PMID- 2979785 TI - Comparative investigation of antipyrine half-life and induction of cytochrome P 450 dependent monooxygenases in rats treated with phenylurea herbicides. AB - The short-term effect of three substituted phenylurea herbicides has been investigated on antipyrine plasma half-life in rats. Isoproturon was ineffective, while chlortoluron and diuron decreased the half-life significantly. There was a close correlation between antipyrine half-lives and induction of hepatic cytochrome P-450 dependent benzo(a)pyrene monooxygenase, 7-ethoxycoumarin O deethylase and 7-ethoxyresorufin O-deethylase. PMID- 2979786 TI - Biological activities and antitumor mechanism of an immunopotentiating organogermanium compound, Ge-132 (review). AB - The biological activities and antitumor mechanism of an immunopotentiator, Ge 132, is reviewed herein. Ge-132 exhibited antitumor activity against certain syngeneic and allogeneic experimental tumors. It was shown that T-cells and macrophages were involved when tumor-bearing mice were protected by the compound. This protective effect could be transferred to tumor-bearing mice, not treated with the compound, by a macrophage fraction and serum specimens obtained from Ge 132-treated mice. Interferon gamma (IFN gamma) was detected in the circulation of Ge-132-treated mice and when sera obtained from Ge-132-treated mice were treated with anti-IFN gamma antiserum in vitro, the antitumor activity was abolished. On the other hand, in mice treated with anti-IFN gamma antiserum, Ge-132 did not induce serum IFN and failed to protect against death due to ascites tumor progression. The in vivo administration of monoclonal anti-Thy 1.2 antibody prevented the expression of the antitumor activity of Ge-132. However, serum specimens obtained from Ge-132-treated mice effectively inhibited the tumor growth of T-cell-depleted mice bearing ascites tumors. Since it has been reported that T-lymphocytes produce IFN gamma, this suggested that Ge-132 may first stimulate T-cells to produce IFN gamma in the expression of the observed antitumor efficacy. In addition, sera obtained from Ge-132-treated mice did not show any antitumor activity in mice depleted of macrophage functions. Additionally, passive transfer of macrophages from mice treated with these serum specimens to tumor-bearing mice also resulted in the inhibition of tumor growth. Pretreatment of these serum specimens with anti-IFN gamma antiserum effectively prevented the generation of cytotoxic macrophages. Also, tumor-bearing mice treated exogenously with this antiserum did not differ significantly in survival as compared to controls, despite the administration of Ge-132. Furthermore, the antitumor activity of Ge-132 was detected in NK cell-depleted mice. Therefore, the antitumor mechanism of Ge-132 in the murine ascites tumor system may be expressed as follows: (a) Ge-132 stimulates T-cells to induce IFN gamma when mice are treated orally with the compound, (b) IFN gamma activates macrophages to become cytotoxic, and (c) the cytotoxic macrophages eliminate tumor cells. PMID- 2979787 TI - In vivo antineoplastic activity of mouse or human IgG conjugated with melphalan. AB - IgG of normal female ICR mice was obtained and covalently conjugated with an alkylating agent, melphalan. The conjugate exhibited marked antitumor activities against Sarcoma 180 and Erhlich carcinoma cells inoculated subcutaneously into ICR mice. Based upon this experiment, melphalan was conjugated with human IgG. Antitumor activities of the conjugate, named K18, were tested against a variety of murine tumor cell lines in vivo. Additionally, K18 was tested in vivo against MX-1, MK-2 and LC-10 human tumor cell lines (of the breast, stomach, and lung respectively). The results showed antineoplastic activities on both murine and human tumor cell lines, thus indicating a possible new antitumor agent. PMID- 2979789 TI - Hypo- and hyperdiploidy in cultured amniocytes. AB - 2,340 metaphases of amniocytes from 149 karyotypically normal women were analyzed for hypo- and hyperdiploidy. Hypodiploidy was more frequent than hyperdiploidy. Chromosome loss was correlated with size. The smaller chromosomes are lost more frequently (r = 0.62 p less than 0.05) than the larger ones. Considering the inconclusive findings of lymphocyte cultures, one can conclude that the hypodiploid cells arose as technical artifacts while hyperdiploidy may be due to somatic non-disjunction owing to pseudomosaicism. PMID- 2979788 TI - Cytostatic and antitumour properties of a new series of Pt (II) complexes with cyclopentylamine. AB - Ten new Pt (II) complexes were synthesized and tested as potential antitumor drugs in vitro on KB human tumour cell line, and in vivo against four experimental tumour systems (P388, L1210, ADJ/PC6A and Yoshida sarcoma). The complexes contained two primary amine ligands (cyclopentylamine) with bidentate leaving ligands consisting of nitro-, dinitro- and sulfo-derivatives of phthalic and isophthalic anions. Various complexes showed a good cytostatic effect in vitro with ID50 from 0.29 and 0.99 mcg/ml. The Pt(cpa)2 (5-sulfo-IPA) appeared to be the most effective compound against P388 and L1210 (T/C% 310 and 250 respectively after three 50 mg/Kg i.p. injections) as well as against ADJ/PC6A (ID90 2.8 mg/kg after a single i.p. injection) and Yoshida sarcoma (T/C% 17.5 after a single 50 mg/kg i.p. injection), but the phthalic acid nitro-derivatives were also quite effective. As far as antileukemic effect was concerned, there was a fairly good correlation between the results in vitro and in vivo. Relationships between antitumour activity and pi electronic charge localization on O- atoms of leaving ligands (M.O. Huckel's Calculations) are also discussed. PMID- 2979790 TI - Relationship between mammary tumourigenesis and uterine adenomyosis in four strains of mice. AB - The development and the progression of uterine adenomyosis were compared with those of mammary tumours in four strains of mice with different mammary tumour potentials. Cumulative mammary tumour incidences were the highest and the lowest in SHN and C3H/He, respectively, and intermediate in SLN and GR/A. However, the growth rate of palpable mammary tumours was higher in SHN and SLN than in GR/A and C3H/He, between which no difference was observed. SHN was also higher than SLN in this parameter. While almost all mice bearing mammary tumours developed adenomyosis in all strains, there was an apparent strain-difference in the progression of this lesion which paralleled well that of mammary tumours. All results indicate the intimate relationship between the development and the progression of both lesions in mice. PMID- 2979791 TI - A new drug-delivery-system of anticancer agents: activated carbon particles adsorbing anticancer agents. AB - A new drug delivery system comprizing activated carbon particles adsorbing anticancer agents has been developed in order to enhance anticancer efficacy on local lesions and to reduce systemic toxicity. The system is designed to release the adsorbed anticancer agent slowly at a designated concentration level at the local site, and to allow the agent to remain for a long time at the local site, with affinity for the lymphatic system and the surface of cancer cells. Through this process, anticancer efficacy is enhanced at the local lesion and systemic toxicity decreases. Because the size of the particles influences the distribution of the agent, size is selected according to both targeted organs, i.e. lymphatic metastases, carcinomatous peritonitis, and administration methods, i.e. intramural, intracavitary, intrabronchial, or intratumoral administration. PMID- 2979792 TI - Identification of quantitative phenotypes in backcross and intercross offspring. AB - The computer program SKUMIX was used to analyze the values of quantitative traits in controlled breeding experiments using inbred rat strains. The SKUMIX program iterated parameters were used to determine the number of expected phenotypes, the values of means and their ratios in the progeny. The computer computed parameters were identical with the results calculated using the real values. The SKUMIX program may be valuable to analyze quantitative data obtained from intercross and backcross breeding experiments yielding overlapping values that cannot be individually assigned to a particular phenotype. PMID- 2979793 TI - Phenotypic alterations of 7,12-dimethylbenz(a)anthracene and N-methyl-N nitrosourea induced rat mammary tumors during repeated transplantations. AB - New transplantable rat mammary tumor lines derived from 7,12 dimethylbenz(a)anthracence (DMBA)- and N-methyl-N-nitrosourea (MNU)-induced carcinogenesis were established and characterized for biological and morphological criteria in the course of multiple tumor generations growing in female or male Wistar rats. DMBA-derived tumors RMT-1, RMT-2 and RMT-3 converted from adenocarcinoma to fibrosarcoma in the early passages. Conversion proceeded earlier in tumors with a well differentiated epithelial component compared to less differentiated ones, providing evidence for clonal spindle cell selection as the most probable mechanism responsible. On the other hand, MNU-derived tumor lines--RMT-4 and RMT-5--maintained histological patterns of parent tumors for a long time, exceeding 20 passages. Analysis of stromal cell tumor component and its interference with developmental sequences of transplantable tumor progression contributes to the explanation of some recent divergent findings (1, 2, 3, 4, 5). PMID- 2979794 TI - Pharmacokinetics and body distribution of amiodarone and desethylamiodarone in rats after oral administration. AB - The pharmacokinetics and body distribution of amiodarone and desethylamiodarone were studied in rats after single oral administration of 100 mg/kg and 200 mg/kg of amiodarone. The time-course of the concentrations of the drug and its main metabolite was determined by high performance liquid chromatography in serum and tissues up to 24 h. The mean absorption half-life of amiodarone was 1.83 h for both dosages and the mean elimination half-life was 15 h after the 100 mg/kg dosage and 105 h after the 200 mg/kg dosage. The mean bioavailability of oral amiodarone ranged from 17% to 60% with an average of 39%. Desethylamiodarone, the major metabolite of amiodarone, was present over the 24 h period of observation in relatively low levels of 30 to 60 ng/ml after the 100 mg/kg dose and 50 to 110 ng/ml after the 200 mg/kg dose respectively, which is circa 4% and 7% of the corresponding parent drug level. Amiodarone is preferentially distributed in decreasing order in lung, liver, thyroid gland, kidney, heart, adipose tissue, muscle tissue and brain. The metabolite desethylamiodarone exhibited a distribution pattern comparable to the parent drug. However, its maximum concentrations in serum and tissues were consistently lower than the corresponding amiodarone concentrations and varied from 18 to 55% (mean 27%), depending on the acute oral dose applied and on the kind of tissue. The amiodarone tissue/serum concentration ratios were high in lung tissue (60-100) and moderate to high in the other tissues except brain (3-60), and indicate an extensive distribution of the drug with the lung as an organ with specific binding sites or uptake mechanisms and adipose tissue as a reservoir with a large storage capacity. The metabolite tissue/serum concentration ratios were very high in lung tissue (500-800), high in renal, thyroid, liver and adipose tissue (80 200) and moderate in the other tissues except for brain (20-60); they indicate a very extensive distribution of desethylamiodarone with, primarily, lung and to some lesser extent kidney, liver and thyroid gland as organs with sites of metabolism and/or specific binding sites or uptake mechanisms and fat as a reservoir for the drug. A marked increase in the accumulation of amiodarone and desethylamiodarone was observed in adipose tissue after chronic oral administration, whereas the rise in kidney and brain was less pronounced and in the remaining tissues it was insignificant. Our data suggest that the rat is a good model for describing the single oral dose pharmacokinetics and body distribution of amiodarone and desethylamiodarone in man. PMID- 2979795 TI - Hormonal regulation of uterine epithelial cell proliferation. I. Effects of estradiol or progesterone administered separately. AB - Using an in vivo tritiated thymidine (3H-TdR) labeling followed by autoradiography, the effects at different times before sacrifice of single or paired injections of 17-beta-estradiol (E2) or progesterone (Pg) at various concentrations were investigated in adult B6D2FI ovariectomized mouse luminal and glandular uterine epithelium. With regard to the luminal epithelium, E2 (0.25, 2.50 or 25.00 micrograms/animal) exerts a mitogenic influence which is dose related. Pg (125, 600 or 5000 micrograms/animal) exerts a significant proliferative effect only at a high dose. With regard to glandular epithelium, E2 and Pg have an almost identical mitogenic influence which is dose-related. A two step Pg administration (2 x 125 micrograms) inhibits cell proliferation in glandular epithelium as compared to the promoting action exerted by a single injection. No refractory response was observed after a two-step E2 administration in either the luminal or the glandular epithelium. These data demonstrate that cell proliferation in the uterine luminal epithelium of an adult B6D2F1 mouse is very sensitive to E2 but not to Pg, whereas these two steroids have a similar mitogenic influence on the cell proliferation of the glandular epithelium. Moreover, a second Pg administration seems to have an inhibiting effect on cell proliferation as compared to that exerted by the first injection performed 12 or 24 hours earlier. In contrast, the repetition of E2 treatment would seem to exert an additive effect on uterine epithelial cell proliferation. In conclusion, epithelial components of the endometrium are complex steroid targets. The two major cell types (luminal, glandular) differ in their abilities to respond to steroid stimulation by entering into DNA synthesis. PMID- 2979796 TI - Hormonal regulation of uterine epithelial cell proliferation. II. Effects of estradiol and progesterone administered in combination. AB - The effects of estradiol (E2) and progesterone (Pg) administered in combination on the cell proliferation of the ovariectomized B6D2F1 mouse uterine epithelium were assessed by thymidine autoradiography. In the luminal and glandular cell populations Pg inhibits the mitogenic effect of E2 when administered simultaneously or 24 hours after E2; when Pg is given 12 hours after E2 it does not block but slows down the proliferative effect of E2, at least at the concentrations studied here. Given prior to E2, Pg rather exerts an additive (synergistic) effect on uterine epithelia. It is concluded that Pg can exert different actions on the E2- mitogenic effect induced at both luminal and glandular levels. Further investigations are needed to understand better the precise biochemical mechanisms involved in the modulating actions of these steroids on the mouse endometrium epithelial cells. PMID- 2979797 TI - Pathobiology of canine cyclic hematopoiesis (review). AB - Canine cyclic hematopoiesis (CH) was first described in Gray Collies as the lethal gray syndrome, and was subsequently shown to be a counterpart of human cyclic neutropenia (CN). The disease is characterized by a recurrent cyclic change in the levels of neutrophils and other blood elements at approximately 12 day intervals. It is caused by an autosomally recessive gene with pleiotropic effects or a CH gene which is closely linked to a gray color gene. The infectious insult on affected animals is periodic but its clinical and pathologic effects are continual and cumulative Affected dogs die after weaning and rarely survive over 6 months of age. There is evidence of immunoregulatory defects in these dogs. Reciprocal bone marrow transplantation indicates that the defect resides in the bone marrow, but the actual site and mechanism of the defect has not been established. The disease in Gray Collies represents a unique model system for studying the mechanism of cyclic hematopoiesis and hematopoietic regulation. Studies of the disease have made conceptual contributions toward understanding and treatment of human cyclic neutropenia. PMID- 2979798 TI - Enhancement of DMBA-induced mammary cancer in Wistar rats by unsaturated fat and cholestyramine. AB - Promotion of 7,12-dimethylbenzantracene (DMBA)-induced mammary cancer in Wistar rats, by diets containing unsaturated fat (USF) and unsaturated fat plus 4% cholestyramine (USF+ CHST) was compared to that in rats fed saturated fat (SF). The diets were fed for 100 or 200 days. Histologic examination of grossly normal mammary tissue, as well as of any tumor mass, showed a significant increase in the incidence and size of mammary tumors in USF and USF + CHST groups when compared to SF group. Moreover, in the USF + CHST group one tumor developed in one rat as early as 100 days. The serum total lipids, triglycerides and cholesterol esters decreased significantly at 200 days in USF and USF + CHST groups. These results suggest that a diet high in unsaturated fat alone, or in combination with 4% cholestyramine, promotes DMBA-induced mammary cancer in Wistar rats. PMID- 2979799 TI - Action of 24R, 25 dihydroxyvitamin D3 (24,25 (OH)2D3) on bone in vivo. AB - The administration of 24R,25(OH)2D3 for 2 years led to an increase in bone volume in the rat. Radiographs showed the striking increase in the density of the bones from animals treated with large amounts of 24R,25(OH)2D3. Total bone minerals measured by direct single photon absorptiometry showed a dose-dependent increase of up to approximately 150% that of controls in both the femur and coccyx of the treated animals. Histological observation revealed a thickened cortex with narrowed medullary area, and Villanueva's bone stain indicated the increase of mineralized bone. The result clearly demonstrates that 24,25(OH)2D3, if massive doses are given, has the biological action of increasing bone volume without disturbing the development of bone sizes. PMID- 2979801 TI - Cell membrane lipid fluidity and receptor expression in Moloney- and Friend virus transformed cells. AB - Normal and malignant cells show differences in cell membrane lipid fluidity (CMF) which influence the expression of membrane receptors and may interfere with cell function. Friend virus (FLV) and Moloney virus (MLV) infected hematopoietic and lymphoid cells were monitored for CMF (fluorescence polarization) and for transferrin (TFC) and thymic (Thy) receptors (FITC-labelled monoclonal antibodies). CMF was modulated with cholesterol hemisuccinate (CHS), phospholipids (PL) and DMSO. Erythropoietic stem cells exhibit an increased persistent CMF within minutes after FLV infection; transferrin receptors are expressed, yet no hemoglobin is synthesized. CHS rigidification reduces TFC expression with differentiation of cells and hemoglobin synthesis, yet transformed cell populations do not react uniformly. Thymic lymphocytes, instead, do not exhibit changes in Thy expression upon CHS treatment although cell membranes become rigidified. Separate experiments showed these cells not being "transformed" per se but blocked in differentiation because of viral destruction of thymic epithelial cells with loss of thymopoietin in vivo. Thus viral cell transformation is followed by non-rigid but persistent membrane fluidization interfering with only selective receptor expression. PMID- 2979800 TI - Multidisciplinary evaluation of rat renal cell carcinoma. AB - The rat renal cell carcinoma system as described by deVere White and Olsson in 1980 is used widely as a model for its human counterpart. The tumor arose spontaneously in a male Wistar Lewis rat and its behaviour has been shown to be stable during multiple passages. We have compared this tumor with the human renal cell carcinoma using a multidisciplinary approach. Light microscopy and electron microscopy showed a great resemblance of this rat tumor to a human renal cell carcinoma of the clear cell type with the ultrastructural presence of desmosomes. With the use of tissue specific antibodies against intermediate filament proteins, it could be shown that their expression is comparable to human renal cell carcinoma, i.e. coexpression of vimentin and different cytokeratins in the tumor cells. The cells could also be shown to contain cytokeratin 18. An aneuploid cell population in the tumor, expressing both vimentin and keratin, could be characterized by DNA flow cytometry in double labeling experiments. Comparison of normal and malignant rat kidney tissue by Northern blot analysis revealed increased levels of vimentin mRNA. In conclusion, this tumor model seems to have several histological and biological properties in common with the human renal tumor. PMID- 2979802 TI - Hypocalcemic effect of 24R, 25-dihydroxyvitamin D3 in rats. AB - The physiological significance of 24R,25-dihydroxyvitamin D3 [24R,25 (OH)2D3] is still controversial. The effects of 24R,25-(OH)2D3 on serum calcium level were examined in rats. While 24R,25-(OH)2D3 had no effect on serum calcium level in normal rats, 200 micrograms/kg of 24R,25-(OH)2D3 showed significant hypocalcemic effect in rats pretreated with vitamin D3 (changes of serum calcium level compared with control group were 1.3 and 1.5 mg/dl at 24 and 36 hours respectively) and also showed a significant antagonistic effect against the action of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) in the thyroparathyroidectomized rats (changes of serum calcium compared with control group were 0.5 and 0.8 mg/dl at 12 and 24 hours respectively). It was concluded that the effect of 24R,25-(OH)2D3 on serum calcium differs from that of 1 alpha,25-(OH)2D3. PMID- 2979803 TI - Microsomal deacetylation of isomeric acetamidobiphenyls. AB - In vitro rates of deacetylation were measured with carcinogenic and noncarcinogenic isomeric arylamides using microsomal preparations from guinea pig, hamster, rat, mouse, rabbit and dog. Following incubation at 37 degrees, the substrate and hydrolysis products were extracted into ether and the corresponding aromatic amine quantified by gas-liquid chromatography. The data show that rates of deacetylation were both species and isomer dependent. In general, the highest rate of deacetylation in microsomes from all species was recorded with 2 acetamidobiphenyl (2-AABP); however, the mouse uniquely deacetylated 4 acetamidobiphenyl (4-AABP) most rapidly. The meta isomer 3-acetamidobiphenyl (3 AABP) was deacetylated at an intermediate rate in all species investigated. The variability in the rates of deacetylation within species may reflect the heterogeneity of the deacetylase enzymes and may be an important factor in amine/amide carcinogenesis. PMID- 2979805 TI - Perspective: growth factors in normal and abnormal fetal growth. AB - Growth factors (GFs) and growth inhibitors (GIs) are important regulators of cell proliferation and differentiation in many cell types. Insulin growth factor I and II, epidermal growth factor, transforming growth factor alpha and transforming growth factor beta are produced and secreted by various fetal tissues, and their cognate membrane receptors are present in the fetal-placental unit. These findings suggest that fetal growth and development are regulated by these or other GFs and GIs in an autocrine or paracrine manner. A well-coordinated and integrated action of several GFs/GIs may be operating in promoting the growth and development of different fetal organs during the various stages of gestation. An interruption of the coordinated pathway due to an abnormal expression of GFs/GIs, a structural aberration of their receptors, or an impairment in the postreceptor signaling mechanism may contribute to abnormal fetal growth. The investigation of the unique role of a single GF/GI, the integrated action of several of them in fetal growth, and the points of abnormalities in this mechanism in abnormal fetal growth have both fundamental and clinical relevance. PMID- 2979804 TI - Functional and morphological alterations following isolated rat stomach irradiation. A model for estimation of radiation injury. AB - Exposure of the stomach to tumoricidal doses of radiation is associated with functional and morphological changes. The experimental data have been mainly obtained from studies of whole body irradiation of the stomach. We investigated the separate effect of irradiation on the surgically exposed stomach with doses of between 3 and 30 Gy. A dose-dependent decrease in free and total acidity was observed with relatively low doses (5-9 Gy). Serum pepsinogen and serum gastrin levels increased following irradiation with doses ranging from 5 to 15 Gy. Two phases of increased regenerative activity were documented (between the 1st and 3rd weeks and between the 4th and 8th weeks). No radioprotective effect was observed when sucralfate or misoprostol were utilized. Further investigations are mandatory in order to find an effective radioprotector for the irradiated stomach. PMID- 2979806 TI - Effects of multiple administration of halothane on the mixed function oxidase system in liver microsomes. Difference between guinea pigs and rats. AB - The effects of multiple administration of halothane on the mixed function oxidase system in liver microsomes were investigated and compared between guinea pigs and rats to clarify the difference in sensitivity to halothane. The mixed function oxidase system, except for NADPH-cytochrome P-450 recductase activity was increased in both animals. The inhibition in guinea pigs may have resulted from the microsomal membrane damage induced by the acceleration of lipid peroxidation. PMID- 2979807 TI - Cellular immune mechanisms in the pathogenesis of Crohn's disease. AB - Crohn's disease is a chronic relapsing inflammatory disease of the intestine of unknown cause. It has been suggested that the disease may result from an abnormality of the immunological functions of the gut. Recent advances in the study of the gastrointestinal immune system show that T cells in the intestinal mucosa are more activated, contain a higher proportion of T4 cells having the phenotypic and functional characteristics of helper-inducer cells, have greater capacity for IL-2 production, and have altered responsiveness to antigen stimulation. In the intestinal mucosa in Crohn's disease the predominance of T cells with helper-inducer function is maintained, and there is no evidence of augmented suppressor activity. Although natural killer cells are infrequent in the intestinal mucosa in Crohn's disease, lymphokine activated killer cell precursors and cytolytic T cell precursors are present and it is possible that these cells also play an important role in the disease. The failure to identify specific infections or environmental etiologies in Crohn's disease is consistent with the hypothesis that the disease is due to an inappropriate immunological hyper-responsiveness to ubiquitous components of the alimentary tract. PMID- 2979808 TI - Myasthenia gravis studied by monoclonal antibodies to the acetylcholine receptor. AB - Myasthenia Gravis (MG) is caused by auto-antibodies to the nicotinic acetylcholine receptor (AChR). Rapid progress has been made recently in the study of both the disease and the AChR. A significant part of this progress can be attributed to monoclonal antibodies (mAbs) raised against the AChR. These antibodies have been used to identify, purify and study the structure, function and biosynthesis of AChR from electric organs, muscles and neurons. In MG, these mAbs have been used as model auto-antibodies in in vivo and in vitro studies. They have also been used as tools for the structural and functional analysis of the patients' antibodies. A region on a small segment of the AChR alpha-subunit (main immunogenic region) dominates the immunogenicity and probably the pathogenicity of the molecule. PMID- 2979809 TI - The nephropathy of systemic lupus erythematosus. AB - Lupus nephritis may be considered the prototypic autoimmune disease which is initiated by immune complex deposition. Several patterns of localization of immunoreactants are observed in lupus nephritis, but few data are available to elucidate the corresponding immunopathogenetic mechanisms. Corticosteroids form the mainstay of therapy for lupus nephritis, provided that they achieve a prompt clinical remission which can be sustained with alternate day therapy. Cyclophosphamide may have fewer risks than extended high-dose prednisone and it has been shown to be more efficacious than corticosteroids in preventing end stage renal failure. PMID- 2979810 TI - Therapeutic strategies in autoimmune disease: cyclosporin A as a model agent. AB - Cyclosporin A is the derivative of the cyclic undecapeptide which is produced by 2 strains of fungi, Cylindrocapon lucidum Booth and Tolypocladium inflatum Gams. The drug has achieved widespread use in the treatment of organ transplant recipients as a vanguard against rejection, and currently is being evaluated as an immunosuppressive agent for the therapy of primary autoimmune phenomena as well. Its mechanism of action, and application to the treatment of rheumatologic disorders will be discussed. PMID- 2979811 TI - Clinical aspects of antibodies to DNA. AB - The detection of antibodies to DNA is one of the most important laboratory tests in rheumatology and immunology from both scientific and clinical points of view. The most useful methods for detection of anti-DNA are the liquid phase radioimmunoassay (Farr assay), solid phase enzyme-linked assays (ELISA) and immunofluorescence (Crithidia Luciliae). The clinical value of detection of anti DNA can be summarized as follows: (a) Antibodies to DNA (particularly those reactive primarily with double-stranded DNA determinants) are highly specific for the disease SLE; (b) Levels of anti-DNA bear a close relation to disease activity in many patients. Rapidly rising levels are frequently associated with a subsequent exacerbation and clinical improvement is often accompanied by declining levels of anti-DNA. However, a minority of patients may have persistent elevations of anti-DNA for extended periods in the absence of overt clinical disease activity; (c) Although a single determination of anti-DNA has little prognostic value, the persistent presence of high levels or the absence of anti DNA may define patient subsets with poor and good prognoses respectively; (d) Antibodies to single stranded DNA although present in some patients with discoid lupus and "ANA-negative" lupus, have little diagnostic specificity and are less valuable for disease follow-up as compared with antibodies to double-stranded DNA. PMID- 2979812 TI - Abnormalities of the immune system in the idiopathic inflammatory myopathies. AB - This brief report reviews some of the major studies presenting evidence for altered humoral and cellular immunity in the idiopathic inflammatory myopathies. Clearly, the association of these disorders with other autoimmune diseases, the myriad of myositis-associated autoantibodies, and the phenotypic and functional abnormalities of mononuclear cells from PM/DM patients, all suggest that autoimmune mechanisms are central and pervasive aspects of these diseases. Yet, the current experimental limitations in immunology, as well as the rarity and heterogeneity of the idiopathic inflammatory myopathies, combine to limit our understanding of which factors are primary, and which are only secondary epiphenomena, in the complex pathogenesis of these disorders. PMID- 2979813 TI - Mechanisms of anti-DNA production in human and murine SLE. AB - Antibodies to DNA occur prominently in systemic lupus erythematosus and play a central role in pathogenesis. Recent studies on the production of anti-DNA antibodies in both mouse and man suggest that this response can result from a variety of immunoregulatory disturbances of B and/or T cells. The immunogenic role of DNA as opposed to other self or foreign antigens remains unclear, however. Future studies will delineate the structure of anti-DNA antibodies and thereby determine the role of antigen drive in this response. PMID- 2979814 TI - Autoimmune endocrinopathies. AB - A great deal of understanding of the human autoimmune endocrinopathies has come from studies of thyroiditis in experimental animals. Three such models are available: induced thyroiditis produced by injections of thyroglobulin plus adjuvant; spontaneous thyroiditis in genetically susceptible animals; and thyroiditis resulting from manipulation of the immunological apparatus. The major lessons learned from studies of experimental animals are (i) autoimmune disease is multifactorial and polygenic; (ii) one or more genes regulating the immune response are associated with the major histocompatibility complex (Mhc) of the particular species; (iii) genetic control of tissue damage is more restricted than control of autoantibody formation; (iv) the Mhc controls T-cell proliferation to the autoantigens; (v) a distinct population of T cells prevents the development of organ-specific autoimune disease; (vi) the suppressor population emigrates from the thymus at a time and rate different from the helper populations; (vii) the suppressor population is more susceptible to low levels of irradiation and to cytotoxic effects of antiserum to Lyt-1 than is the helper population. PMID- 2979815 TI - Cellular immunity in systemic lupus erythematosus. AB - Production of pathogenic autoantibodies in patients with systemic lupus erythematosus (SLE) is the cornerstone in the pathogenesis of the disease. From the etiologic point of view, there is evidence that genetic, infectious, hormonal and environmental factors are essential to the development of SLE. The relative contribution of each group of factors to the expression of disease is individualized to each patient or group of patients with identical clinical disease. The cellular events which precede the B cell overactivity are crucial. T cells may produce excessive amounts of factors promoting B cell function, decreased amounts of suppressive factors or diminished cytotoxic ability to eliminate autoreactive T or B cells. The current understanding of lymphocyte and macrophage abnormalities are presented critically in this communication. PMID- 2979816 TI - Autoantibody production in Sjogren's syndrome: a hypothesis regarding defects in somatic diversification of germ line encoded genes. AB - Patients with primary Sjogren's syndrome (SS) have rheumatoid factor (anti-IgG Fc antibody, RF) in their sera that contains a crossreactive idiotype (CRI) defined by monoclonal antibody 17-109. This CRI is shared by SS patients and certain patients with Waldenstrom's macroglobulinemia. This CRI was not found in increased frequency in RF from rheumatoid arthritis patients lacking SS. The structural basis for this CRI is the highly conserved kappa chain of the Vk IIIb sub-subgroup. In SS patients, the CRI is present on both IgA RF and IgM RF, accounting for 5-20% of the total RF. Recent studies have shown that a germ line encoded kappa variable region gene (Humkv 325, cloned from a placental DNA library) has DNA sequence that encodes the variable region amino acid sequences of CRI positive kappa chains. Using MoAb 17-109, we have been able to determine the tissue distribution of CRI+ B-cells. We found an increased frequency in the salivary gland biopsies of SS patients and also in their intestinal mucosal tissues (i.e. Peyer's patch). In comparison, we also detected an increased frequency of CRI+ B-cells in the Peyer's patch regions of normals. Taken together, these results suggest that CRI+ B-cells may play an important role in normal mucosal immunity and these CRI+ B-cells have migrated to the salivary gland in SS patients as part of the inflammatory process directed at autoantigens or exogenous antigen(s) located at a mucosal surface.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2979817 TI - Autoimmune aspects of human infertility. AB - Both men and women make antibodies against human sperm. For a complete diagnostic evaluation of an infertile couple, we must test the two serum samples, plus her cervical mucus and his sperm cells. The antibody factor occurs in 9% of men and in 12-15% of women from infertile couples, and at various titers. The standard, or classical, methods for antibody detection in serum include several agglutination methods: Gelatin Agglutination Test (GAT), Tumbe-Slide Agglutination Test (TSAT), Tray Agglutination Test (TAT), and Capillary Tube Agglutination Test (CTAT), plus the immobilization (Isojima). There are also newer methods; passive hemagglutination, radiolabel-antiglobulin, ATR-release cytotoxicity, hemadsorption, indirect immunobead, and ELISA. The genital secretions must also be studied. Sperm cells are judged to be antibody-coated by use of the Mixed Anti-Globulin Reaction (MAR) test or the Immunobead Test (IBT). Cervical mucus is dissolved with bromelin and tested. PMID- 2979818 TI - Autoantibodies to DNA. PMID- 2979819 TI - Lymphocytotoxic antibodies. AB - LCTA are a heterogenous group of antibodies found in many disease states. The recurring observation is that they are able to modulate immune function in vitro. The exact mechanism of this modulation is unknown, however, alteration of cell surface antigens appears to be important. Recent advances in molecular biological techniques have made the surface antigen, against which LCTA is directed, more accessible. Once the surface antigen is identified, the exact role of LCTA may then be elucidated. Endothelial cell damage is a recurring pathologic finding in many of the diseases discussed. Again, identification of the antigen and its similarity to antigens on various cell types may help explain the immunopathogenesis of the different disease processes. PMID- 2979820 TI - Rheumatoid factor diversity. AB - Rheumatoid factors, antibodies directed against the Fc portion of IgG, exhibit considerable isotypic and idiotypic diversity. Differential expression of IgM and IgA rheumatoid factors suggests that isotype-specific regulatory pathways are operative in man. The physical characteristics of IgA rheumatoid factor in rheumatoid arthritis have also been investigated. Although the polymeric form of IgA rheumatoid factor generally predominates in sera and synovial fluids of rheumatoid arthritis patients, wide variations in the ratio of monomeric to polymeric IgA rheumatoid factor are observed. Both forms are elaborated by synovial plasma cells. That differences in the proportions of monomeric and polymeric IgA RF present in sera may be of pathogenetic significance is suggested by evidence that complexes of polymeric IgA rheumatoid factor and IgG are potent inducers of lysosomal release by RMN whereas monomeric IgA rheumatoid factor-IgG complexes are essentially inactive in this regard. The idiotypic diversity of rheumatoid factors has also been under intensive investigation. A monoclonal antibody designated 6B6.6 recognizes a Vk-associated idiotype present on approximately one-third of IgMk rheumatoid factor paraproteins, but only a small fraction of polyclonal IgM 6B6.6-defined idiotype is distinct from previously described rheumatoid factor cross-reactive idiotypes. Taken together, these studies underline the extensive diversity of rheumatoid factors present in various conditions. Application of molecular genetic approaches to the study of rheumatoid factors should provide explanations for the molecular basis of this diversity and serve to clarify the relationship among rheumatoid factors expressed in both physiologic and pathologic states. PMID- 2979821 TI - On the origin and control of C3NeF. AB - Nephritic factors are a relatively recent addition to the growing list of autoantibodies. C3NeF was the first of these to be described and acts by stabilizing the alternative pathway C3/C5 convertase. IgG and IgM C3NeF as well as anti-idiotypic antibody to C3NeF, may be found in normal individuals at all ages. In addition, internal image anti-idiotypic antibody can also be demonstrated in patients with glomerulonephritis. Thus, C3NeF appears to fulfill all of the criteria for being a product of normal B-cell function. The potential dysregulation of this process and the role of C3NeF in glomerulonephritis remains theoretical at best. PMID- 2979823 TI - Pathogenesis of uveitis. PMID- 2979822 TI - Insulin dependent diabetes mellitus as an autoimmune disease. AB - In this review we have summarized several lines of evidence supporting the concept that insulin dependent diabetes (IDDM) is an autoimmune disease. Initially, we have considered the association of IDDM with other autoimmune diseases, a fact that indirectly suggests involvement of the immune system. Other evidence implicating the immune system is the presence of auto-antibodies directed against pancreatic islet cells and cellular components of the immune system associated with pancreatic insulitis. Finally, we have examined the reported association of cell-surface proteins named histocompatibility antigens (HLA) with this particular disease. Structural and functional aspects of HLA have been considered along with regulatory mechanisms concerning the expression of these antigens. PMID- 2979824 TI - Autoimmune aspects of aplastic anemia. AB - Recovery of bone marrow function in aplastic anemia patients treated with immunosuppressive therapy first suggested a role for the immune system in bone marrow failure. High recovery rates in patients treated with immunosuppressive therapy suggested that an immune mechanism may be a final common pathway of marrow failure in this disease. In vitro studies have shown that aplastic peripheral blood and marrow cells and their supernatants are capable of suppressing hematopoiesis by autologous and normal marrow. Soluble factors identified in this system include gamma interferon and lymphotoxin. The interaction of these molecules with positive growth factors, the role of synergy with other negative regulators, and their role in the pathogenesis of bone marrow failure are discussed. Lymphokine and lymphocyte abnormalities in aplastic anemia may be manifestations of an underlying viral etiology. Three examples are discussed: Epstein-Barr virus-associated aplastic anemia; B19 parvovirus bone marrow failure; and HIV-induced neutropenia. PMID- 2979825 TI - The pathogenesis of rheumatoid arthritis. AB - The results of recent immunohistological studies and ex vivo and in vitro experiments in rheumatoid arthritis (RA) are presented. These findings are used as the basis for a unifying concept of the pathogenesis of RA which serves to explain many clinical and laboratory features of this disease. This hypothesis, which may be regarded for experimental purposes as being analogous to the in vitro AMLR, provides a rational basis for the further analysis of the cellular immune mechanisms operating in RA and the development of effective therapeutic regimens. The ultimate goal is the early diagnosis and cure of RA before the development of irreversible joint damage. PMID- 2979826 TI - Sex differences in adrenocortical structure and function. XXVI. Further studies on the differences in the compensatory growth of the adrenal cortex of the male and female hamster after unilateral adrenalectomy. AB - The relative adrenal weight of the remaining right adrenal gland was significantly increased 5 days after unilateral adrenalectomy in hamsters of both sexes. This effect was associated with an enlargement of the zonae glomerulosa and fasciculata in males and of the zona fasciculata in females. Unilateral adrenalectomy raised the average volume of zona fasciculata cells in male hamsters and that of zona reticularis cells in females. The number of parenchymal cells in all adrenocortical zones remained unchanged by the removal of the contralateral gland; however, when calculating per mg of adrenal or per cell, the nucleotide uptake by adrenal quarters was notably lowered. A marked drop in plasma cortisol concentration was found in unilaterally adrenalectomized male but not in female hamsters. These findings indicate that the main component of the adrenal compensatory growth in hamsters is the hypertrophy and not the hyperplasia of parenchymal cells. PMID- 2979827 TI - Flow cytometry in invasive endometrial carcinoma. Correlations between DNA content S-phase rate and clinical parameters. AB - In a prospective study including 185 women with endometrial adenocarcinoma, the DNA-content and the S-phase rate were measured by flow cytometry. The tumors of 138 (75%) patients were peridiploid and those of 47 (25%) were grossly aneuploid. Aneuploid tumors were more common in poorly differentiated tumors than in well and moderately differentiated tumors (p less than 0.0001). S-phase rates were evaluable in 148 (80%) tumors. High S-phase rates were more frequent in stages III + IV than in stages I + II (p less than 0.05) and were also more common in poorly differentiated tumors than in moderately and well differentiated tumors (p less than 0.05). Ploidy levels and S-phase rates were not significantly correlated with age or menopausal status. During the follow-up time (6 to 38 months), 19 patients died of their disease. The number of deaths registered was significantly higher among patients with aneuploid tumors than among patients with peridiploid tumors (p less than 0.002). The proportion of tumors with high S phase rates was significantly higher than that of tumors with low S-phase rates (p less than 0.0001). These preliminary results indicate that patients with aneuploid tumors or tumors with high S-phase rates are at high risk for early recurrences. PMID- 2979828 TI - Antitumor effect of 24R,25-dihydroxyvitamin D3. AB - 24R,25(OH)2D3, one of the endogenous active metabolites of vitamin D3, showed suppressive effects on the proliferation of various tumor cells in vitro and a prolonging effect on the survival of P-388 bearing mice in vivo. Lewis lung carcinoma was found to cause hypercalcemia in tumor bearing mice. K-DR (24R,25(OH)2D3 (prepared by Kureha Chemical Ind.) significantly prolonged the survival of mice with Lewis lung carcinoma. K-DR also showed a suppressive effect on the growth of human osteosarcoma transplanted in nude mice. PMID- 2979829 TI - Evaluation of retinyl palmitate and cytarabine treatment in a slowly growing rat leukemia model. AB - The slowly effect of retinyl palmitate in prolonging survival time in a slowly growing rat promyelocytic leukemia model (BNML) was tested. Retinyl palmitate was given in addition to cytarabine, which has previously been shown to prolong survival time in these rats. The effect of cytarabine on liver store and plasma level of retinol was also tested. The results show that retinyl palmitate did not prolong survival time in the leukemic rats. Similarly, cytarabine did not reduce the level of retinol in liver or plasma. PMID- 2979830 TI - DNA synthesis in the gastroduodenal mucosa during acute and chronic stress in the rat. AB - We investigated the effects of acute and chronic stress on the DNA synthesis of the gastroduodenal mucosa of the rat using two different methods of physical stress at various time intervals. Acute stress was produced in the rats being briefly plunged or swimming for two hours (water temperature 37 degrees C). "Sham - transported" rats were used as controls. The results indicate that in the stomach the DNA synthesis was substantially reduced during acute stress in both groups tested (when compared to controls). The DNA synthesis was also reduced in experimental rats after one and two weeks of stress (as compared to day one). By four and eight weeks, the rate of DNA synthesis in the gastric mucosa had significantly increased in the stressed animals. Controls demonstrated significantly lower DNA values following two to eight weeks of stress (as compared to day one). From the outset, the DNA replication values were 2.5 to 3 times higher in the duodenal mucosa than in the gastric mucosa. Following two weeks of stress, the duodenal mucosa of both test groups showed significantly lower DNA values than controls, but significantly higher values after four weeks of stress. By eight weeks, the duodenal mucosa in all rats had reached the same values as that of day one. This was considered a sign of "adaptation to stress" in the duodenal mucosa. The above results suggest that the fluctuations of DNA replication may be connected to compensatory mechanisms aimed at adjusting the gastroduodenal mucosa to protracted stress situations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979831 TI - In vivo antineoplastic activity of ascorbic acid for human mammary tumor. AB - The effect of ascorbic acid on the growth of human mammary tumor xenografts was investigated using the 6-day subrenal capsule assay method. The results showed that ascorbic acid (1 or 5 g/liter) administered in the drinking water significantly inhibited the growth of tumor fragments implanted beneath the renal capsule of immunocompetent mice. The results agree with other work carried out in animal experiments with animal tumors. Administration of ascorbic acid in the mouse diet did not affect the growth of the human mammary tumor fragments within the 6-day experimental period. Tumor growth was inhibited when mice were fed a diet containing ascorbic acid (50g/kg diet) together with cupric sulfate (18 or 90 mg/liter) in the drinking water. The results support the hypothesis that certain oxidation and degradation products of ascorbic acid are active antineoplastic agents for the human mammary carcinoma studied. PMID- 2979833 TI - Single-chain, urokinase-type plasminogen activator in a tumor model linked to metastatic potential. AB - Single-chain urokinase-like plasminogen activators have been determined by an indirect method (after activation to urokinase by plasmin and chromogenic assay with S 2444) in the cytosol of a rat tumor model. Variants of the R 3230 AC rat mammary adenocarcinoma were studied, including a subline with increased metastatic potential established by lung colony assay. The cellular content of the UK precursor was found to be significantly higher in the metastatic variant. Plasminogen activator-mediated invasion and collagenolysis of cells with the metastatic phenotype may be regulated by the release of the precursor which lacks reactivity with inhibitors, its transient activation and subsequent inhibition of the urokinase formed. PMID- 2979832 TI - Comparison of tumor growth inhibitory and toxic effects of a new fluorouracil- nitrosourea derivative (B-3839). AB - The toxic effect and anti-tumor activity of B-3839, a new molecular combination of pyrimidine antimetabolite 5-fluorouracil (5-FU) with the alkylating agent N Chloroethyl-N-nitrosourea (BCNU), was compared to that of BCNU and 5-FU given alone and in physical combination. The tumor inhibitory effect of B-3839 was similar to that of BCNU given alone or combined with a low dose of 5-FU in the i.m. Walker tumor model. Furthermore, the bone marrow toxicity of BCNU was not significantly altered by either form of combination with 5-FU. The intestinal side effects, evaluated by measuring the decrease of marker enzyme (thymidine kinase, xanthine oxidase, alkaline phosphatase, sucrase, maltase) activities in isolated enterocytes, were dose-dependent and moderate. A significant, more than 30%, decrease occurred only if BCNU and 5-FU were given simultaneously or as B 3839. The molecular combination of the two drugs does not provide any additional advantage over their physical combination. PMID- 2979834 TI - Effects of single and combined maltose tetrapalmitate immunotherapy, cyclophosphamide chemotherapy and radiotherapy on ethyl carbamate accelerated primary lung cancer in A/J mice. AB - A/J mice were given ethyl carbamate to accelerate and to raise to 100 percent the incidence of lung tumours at 34 weeks (day 237) of age. The animals were then divided into groups which received the following treatments: group 1, no treatment; group 2, MTP alone; group 3, radiotherapy alone; group 4, cyclophosphamide alone; group 5, radiotherapy + MTP; group 6, MTP + cyclophosphamide; group 7, radiotherapy followed by cyclophosphamide and group 8, MTP and radiotherapy together followed by MTP and cyclophosphamide. Except for radiotherapy, which was given for 5 consecutive days, MTP and cyclophosphamide were continued till the death of the animals. The treatment efficacies were evaluated by the number and size of tumour nodules, taking into consideration the survival time of the animal. Animals in groups receiving cyclophosphamide died earlier (between days 290 and 315) due to its toxic effects, and half of the radiotherapy-MTP were sacrificed at day 314 for comparison. Although cyclophosphamide alone and radiotherapy plus cyclophosphamide demonstrated antitumour activity, the number of tumour nodules and the nodule diameter were reduced most effectively in group 8 (receiving MTP, radiotherapy and cyclophosphamide). Among the animals in the non-cyclophosphamide group, radiotherapy alone was ineffective. MTP given before and after radiotherapy (group 5) kept tumour volume in control although this group died suddenly. The animals receiving only MTP died between day 430 and 470. The number of tumour nodules and the nodule diameter in the MTP group were, however, significantly reduced when compared to controls or radiotherapy group animals dying at or near the same time. PMID- 2979835 TI - Difference in response to mammotropic hormones of mammary growth and uterine adenomyosis among four strains of mice. AB - The effects of single pituitary grafting and treatments with progesterone or oestrogen on mammary gland growth and uterine adenomyosis were studied in four strains of female and male mice with different mammary gland growth potentials (SHN/Mei, SLN/Mei, GR/AMei and C3H/HeMei). The order of responses of different strains of mammary gland growth to hormone manipulations generally parallelled those in the intact controls, while there was some strain - related difference in response to individual hormones. The relationship between the experimental and the control groups seen in mammary gland growth was not always observed in the development and the progression of uterine adenomyosis in all strains. These results indicate that the importance of cell susceptibility to hormones in the growth is different between target tissues. PMID- 2979836 TI - Comparative histochemical study of adriamycin and adriamycinol in rat liver and heart. AB - Investigations were carried out into morphological differences between the uptake of adriamycin and its 13-OH metabolite adriamycinol into rat liver and heart. In the liver parenchyma, adriamycinol is mainly stored in the central area of the hepatic lobule, while adriamycin shows a more uniform distribution. In cardiac tissue, adriamycin accumulates within chromatin masses, whereas adriamycinol scarcely enters myocardial cells. The cardiotoxicity of both anthracyclines might be related to interactions with cytomembrane binding sites. PMID- 2979838 TI - Toxicities of hydrazines: a review. AB - The toxic properties of sixty carcinogenic hydrazines are reviewed and presented. Many of these chemicals are synthesized by man and used in industry, agriculture and medicine, while others occur in nature as ingredients of fungi and plants. The human population is, therefore, exposed to them to a considerable degree. These compounds, in addition to being carcinogens, have toxic effects in the gastrointestinal tract, respiratory, nervous, lymphoreticular, hematopoietic, cardiovascular, genitourinary, integumentary, musculoskeletal systems, etc., of 15 animal species and man. Because they caused toxic damage in practically every organ and tissue, earlier claims concerning their tissue specificity are discounted. Their hazardous properties deserve careful attention. PMID- 2979837 TI - Depression factors of neutrophil bactericidal activity with nitroblue tetrazolium reduction test in surgical patients. AB - In this study, background factors were determined in 108 surgical patients who showed depression in phagocytic and bacteriocidal activity with nitroblue tetrazolium reduction test. Control subjects consisted of 4 healthy males and 4 healthy females whose age ranged from 25 to 38 years. The background factors associated with the depressed neutrophil phagocytic and bacteriocidal activity in surgical patients were renal insufficiency, liver cirrhosis, hypoproteinemia, diabetes mellitus, long-term administration of steroids and immunosuppressants, obesity, anemia, aging and malignant tumors. These depression factors closely resembled those generally considered to be involved in increased susceptibility to infections. PMID- 2979839 TI - Residual myometrial invasion after intracavitary irradiation of endometrial adenocarcinoma stages I and II. Relations to DNA content and S-phase rate. AB - The DNA content and the S-phase rate were determined by flow cytometry in 129 women with endometrial adenocarcinoma in stages I and II. Aneuploidy and elevated S-phase rates were statistically correlated to loss of histopathologic differentiation. After preoperative intracavitary irradiation and surgery, the occurrence of residual myometrial invasion was studied. Residual myometrial invasion was found in 28 (22%) cases. In 17 (17%) of the peri-diploid (1.8c-2.2c) and in 11 (38%) of the aneuploid tumors, respectively, the myometrium was invaded (p less than 0.05). No statistical correlation was found between mean S-phase rate and myometrial invasion. The difference in residual myometrial invasion found between aneuploid and peri-diploid tumors after intracavitary treatment may reflect a primary difference in the tendency to invade the myometrium or a difference in responsiveness to radiotherapy. PMID- 2979840 TI - Antitumor activity of ascorbic acid in combination with antitumor agents against Lewis lung carcinoma. AB - Antitumor activity of ascorbic acid in combination with antitumor agents (mitomycin C and 5-fluorouracil) was examined against subcutaneously implanted Lewis lung carcinoma-bearing C57BL/6 mice by feeding them an ascorbic acid deficient diet. The mice were divided into four groups: group 1 received intraperitoneally mitomycin C (2 mg/kg) and 5-fluorouracil (50 mg/kg) once a week for four weeks beginning from the day after implantation of tumors, as well as ascorbic acid (1000 mg/kg) twice a week for the same four weeks; group 2 received only mitomycin C and 5-fluorouracil; group 3 received only ascorbic acid; group 4 received a vehicle (physiological saline). Tumor growth of group 1 compared with the other three groups, and that of group 2 compared with groups 3 and 4, was significantly inhibited by day 13 post implantation. Histological examinations of tumor tissues at 10 days after implantation of tumors already showed degenerative changes which indicated these antitumor effects. PMID- 2979841 TI - Effects of antiepileptics on the hepatic storage of retinol. AB - The effect of various antiepileptics on the retinol storage in rat liver was tested. We observed a dose- and time-dependent reduction in the hepatic storage of retinol after the administration of several of these drugs. Administration for 11 weeks of phenobarbital, primidone and carbamazepine in doses comparable to those used in humans reduced the retinol concentration in the Liver by 17-33% compared to control rats. The plasma retinol levels remained unaffected in all the rats. Plasma retinol from 31 epileptic children had plasma levels between 0.7 and 2.4 nmol/ml, which is regarded as normal. PMID- 2979842 TI - Chromosomal and molecular basis of human neoplasia (review). PMID- 2979843 TI - A review of experimental adenocarcinoma in the intact and post-operative rat glandular stomach with particular reference to mucin histochemistry. AB - The majority of experimental studies on the development of gastric adenocarcinoma have been performed in the rat. There is evidence for two pathways of carcinogenesis in the intact stomach following carcinogen administration. The first has a sequence of hyperplasia, dysplasia, carcinoma with a benign proliferating phase associated with damage and repair mechanisms. The second sequence progresses through increasing grades of dysplasia in an undisturbed mucosa leading to carcinoma formation. Early experiments concerning the effects of gastric surgery on carcinogen induced adenocarcinoma may have simply altered the effectiveness of the carcinogen. Recently it has been shown that surgery alone can induce adenocarcinomas and the number of tumours is related to the degree of duodenogastric reflux. The component of that reflux which seems to be responsible is pancreaticoduodenal secretions and not bile. Intestinal metaplasia is not an important intermediate stage in carcinogenesis in the rat. PMID- 2979844 TI - Discrepancy between in vitro and in vivo passaged U-937 human leukemic cells: tumorigenicity and sensitivity to differentiating drugs. AB - The treatment of nude mice bearing tumors of transplanted human leukemic cells with drugs known to induce differentiation of the same leukemic cells in vitro does not always affect tumor yield, tumor cell differentiation or nude mice survival. We have transplanted human monoblastic leukemic cells of the U-937 cell line into newborn Swiss nu/nu mice. Priming with cyclophosphamide, followed by subcutaneous injections of at least 10 x 10(6) cells allowed us to obtain solid tumors. The cytology, HLA phenotype and in vitro proliferation characteristics of the U-937 tumor cells were conserved. However, these tumor cells were more tumorigenic when reinjected into nude mice and showed a modified response to differentiation induction. A decreased capacity to differentiate with retinoic acid (RA) and a resistance to 1-beta-D arabinofuranosyl cytosine (Ara-C) and 1-25 dihydroxy vitamin D3 (1-25 (OH)2 D3) were noted in three tumor cell lines tested. With regard to the latter, the resistance was not due to a modification of the number of cell receptors. The study shows that though in vivo transplantation of human leukemic cells in nude mice may lead to a selection of resistant cells, systematic checking of in vitro differentiation characteristics of the tumor cells permits the nude mouse model to be maintained for the in vivo screening of new differentiating agents. PMID- 2979845 TI - The effects of ionizing irradiation on production of thromboxane and prostacyclin by the isolated perfused rat kidney. AB - Exposure to whole body radiation is associated with prompt changes in urinary excretion of prostaglandins. We investigated the separate effects of radiation on rat kidney capillary and tubular system prostaglandin synthesis. Animals were irradiated to the left kidney area with a single dose of 15 Gy. One week later the rats were anesthesized, the renal artery, vein and ureter of the left kidney cannulated, and the kidney removed and perfused with Krebs-Henseleit physiological buffer at a rate of 10-12 ml/min. Effluent fluids were collected separately from the renal vein and from the ureter of irradiated and control (operated sham-irradiated animals) and were assayed by radioimmunoassays for thromboxane A2 (TXB2) and prostacyclin (6 keto PGF1 alpha). Histological examination of the irradiated kidneys showed no significant changes, and electron microscopy revealed minimal interstitial edema. In contrast to these minimal changes, TXB2 assays showed a significant increase both in the venous and ureter effluents. Following stimulation with angiotensin II in the perfusate, a further significant increase in TXB2 production was observed both by the capillary and the tubular systems. With 6 Keto PGF1 a slightly different response was seen. The basal production was increased only in the ureter effluent of the irradiated animals, while there were no changes in the release in the venous effluents. In parallel, radiation significantly increased the angiotensin II stimulated production capacity of prostacyclin by the tubular system. The response of the capillary system following irradiation may create imbalance between these two important substances and lead to the radiation effects in the renal tissue. PMID- 2979846 TI - Polyenoic fatty acid deprivation of juvenile cats modulates 3H-dopamine release from presynaptic receptors in caudate slices. AB - A classical neurotransmitter release model was used to investigate the effect of the limitation of dietary polyenoic fatty acids on the function of presynaptic dopamine autoreceptors in slices from the caudate nucleus of kittens (juvenile domestic cats). Maximum inhibition of 3H-dopamine release was detected only in the brains of animals receiving post delta-6-desaturation fatty acids of both the omega 3 and omega 6 series. The removal of either, or both, groups of fatty acids resulted in attenuation of sensitivity of the autoreceptor to apomorphine. It would appear that omega 6 fatty acids are more important for a normal pattern of dopaminergic function to be established in the caudate nucleus of kittens. PMID- 2979847 TI - Tamoxifen tolerance and steroid receptors in a tumor model. AB - In a rat mammary adenocarcinoma model, prolonged exposure to the antiestrogen Tamoxifen results in a Tamoxifen-tolerant tumor cell line which is readily transplantable and grows under continuous oral intake of the drug. The solid tumor contains steroid receptors; however, the level of Tamoxifen needed for effective displacement of diethylstilbestrol cannot be achieved with therapeutical doses; resistance to Tamoxifen is not the result of diminished or absent estrogen receptors. PMID- 2979848 TI - Use of a specific monoclonal antibody for studying the liver metastatic invasion of a rat colon cancer. AB - A simple model for liver metastasis from colon cancer resulted from the intraportal injection of 2 x 10(7) highly tumorigenic DHD/K12/PROb cells into syngeneic BDIX rats. Early detection and development of cancer invasion were studied by conventional microscopy and immunoenzymatic staining using a specific monoclonal antibody. Metastases developed either from isolated cancer cells early disseminated in sinusoid network or from intraportal microthrombi. An intense immune reaction developed until day 15 after injection but decreased and disappeared at the latest stages of evolution. PMID- 2979849 TI - Hyperthermia or BCNU alone and combined in BD IX rats with BT4An tumours. Effects of a single or two equal fractions at different intervals. AB - BD IX rats with BT4An tumours subcutaneously on the hind leg were stratified after tumour size and treated by single dose 44 degrees C water bath hyperthermia for 90 minutes or with two 45 minute sessions at 24, 48 or 168 hour intervals. Other groups were treated by single dose BCNU at 20 mg/kg or by two divided doses at the same intervals, or by combined hyperthermia and drug as single treatment or as divided treatments at the same intervals. Intratumoural temperatures in some animals were carefully scanned for minimal temperature during two hyperthermia sessions 24 hours apart. Tumour response, normal skin damage and weight was evaluated. It was concluded that single dose hyperthermia and combined treatment yielded the best effect, but also resulted in the most severe skin reaction and weight loss. Hyperthermia was least effective at a 24 hour interval, indicating thermotolerance to be greatest at this time. The minimum intratumoural temperatures were higher at the second heat dose given at 24 hours, demonstrating a poorer ability to dissipate heat after the first treatment. The tumours were only slightly sensitive to BCNU alone, and no optimal schedule was found. BCNU increased the hyperthermia effect at all schedules, except for the 168 hour interval. The skin reaction to single dose hyperthermia was enhanced by BCNU. PMID- 2979850 TI - Fine needle biopsy as a method for following ploidy changes in the Dunning R-3327 rat prostate tumour. AB - The use of fine needle biopsies for the collection of cells for flow cytometry was investigated using the Altman Dunning R-3327 rat prostate tumour. Five rats were castrated 150 days after tumour implantation. Multiple fine needle biopsies were performed 138, 194 and 220 days post implantation and the results compared to those from intact animals. A small aneuploid peak was found at the 2.8c position in intact animals. Although tumour growth was slowing down in castrates, indicating hormone independency but sensitivity, no change in flow cytometric profile could be demonstrated. This indicates that, in this model, no correlation between aneuploidy and hormone independence exists. The fine needle biopsy method proved to be easy and reliable for tumours of above 1 cm3 in volume. PMID- 2979851 TI - Developmental activity profile of DNA polymerases delta and alpha in rat neurons suggests a coordinated in vivo function. AB - DNA polymerase delta, a fourth cellular DNA polymerase, might play an important role in cellular events. The properties of the enzyme suggested possible roles in nuclear DNA replication and DNA repair. By using specific assays that allow the determination of DNA polymerases delta and alpha in crude enzyme fractions (1), we show here that in developing rat neurons, which stop dividing before birth, both DNA polymerases delta and alpha drop sharply in an identical pattern from a high level with the approach of term and disappear at approximately three weeks of postnatal age. The results suggest that DNA polymerases delta and alpha might have a coordinated function during DNA replication in eukaryotic cells. PMID- 2979852 TI - Clinical value of immunolymphscintigraphy in patients with breast cancer. AB - Thirty four patients suspected of suffering from breast cancer were investigated by immunolymphscintigraphy (ILS) a few days before a planned operation. Patients received 500 micrograms of the BCD-F9 monoclonal antibody or its F(ab')2 fragments containing 1 mCi of Iodin-123. Each preparation was given by a subcutaneous injection into the fingerwebs between the 2nd and 3rd fingers of both hands. Three static scintigrams were taken 4, 8 and 24 hours following injection of the radiolabelled antibodies. ILS results were always compared to histopathological findings and gave for the intact antibody a sensitivity of 83%, a specificity of 93% and an accuracy of 89%, the positive predictive value was 91% and the negative predictive value was 80%. On the other hand, when the F(ab')2 fragments were used the sensitivity of the ILS technique was 75%, the specificity was 100% while the accuracy was 86%, the positive predictive value was 100% and the negative predictive value was 75%. PMID- 2979853 TI - Antitumor efficacy of two kinds of tumor vaccine modified with vaccinia virus. AB - We compared antitumor effectiveness of two types of tumor vaccine modified with vaccinia virus (VV). One type is UV-inactivated VV (UV-VV) -absorbed tumor cell vaccine (UV-VV TCV) produced by tumor cells that had been absorbed in vitro with UV-VV and subsequently X-irradiated with 10(4) rads. The other type is vaccinia oncolysate vaccine (VOV) from tumor cells that had been infected in vitro with live VV and subsequently sonicated. C3H/HeN mice were inoculated i.p. with UV-VV after whole body X-irradiation with 150 rads. After 3 weeks, the two kinds of vaccine were administered i.p. 3 times at weekly intervals. One week after the last injection, mice were challenged i.p. with syngeneic MH134 or X5563 viable tumor cells at different doses. The fifty percent tumor lethal doses (TLD50) of MH134 in mice treated with UV-VV TCV and VOV were 10(6.6) and 10(5.3), respectively, whereas the TLD50 of MH134 in non-treated mice was 10(0.6). The TLD50 of X5563 in mice treated with UV-VV TCV and VOV were 10(6.5) and 10(4.4), respectively, while the TLD50 of X5563 in non-treated mice was 10(0.5). These results show that UV-VV TCV is more effective than VOV. We suggest that the complete cell structure of the vaccine is more effective for enhancing tumor immunity. PMID- 2979854 TI - Effect of ELF pulsed magnetic fields on survival of leukaemia-prone AKR mice. AB - In a previous experiment an increase in the survival time of leukaemia-prone AKR mice was observed by exposure to a 600 mT or 800 mT static magnetic field when the mice were at least 200 days old. In this experiment 200-day-old-mice were exposed to a 6 mT pulsed magnetic field (PMF) for 30 minutes a day, twice a week until death. The frequency of the field was 12 Hz or 460 Hz. The exposed mice died from leukaemia but had an increased survival time; the average increase was 14.25% compared to the controls. Both the frequencies gave similar results. PMID- 2979855 TI - Quick freezing of mouse embryos: freezing of inbred strains and 2- and 4-cell embryos by vitrification. AB - Murine embryos of mice of four different inbred strains and one hybrid strain were evaluated for their ability to survive quick freezing by post-thaw in vitro development. The embryos were transferred to an equilibration medium [10% 1,2 propanediol and 20% glycerol in modified PBS (mPBS)] for 10 minutes and frozen in a vitrification medium (25% glycerol and 25% 1,2-prapanediol in mPBS) by direct lowering into liquid nitrogen. Following thawing at 30 degrees C, dilution in 1 M sucrose in mPBS and washing in mPBS the embryos were cultured, and development was evaluated 24-28 hours later. The number of fertilized eggs obtained by superovulation differed among the strains. The survival rates evaluated by in vitro cultivation of the post-thawed inbred embryos varied from 50-85% depending on the genotype, whereas the normal live offspring from transfer of frozen-thawed embryos to recipient females confirms that the quick freezing method is an applicable method for storage of genetically defined mouse strains and stocks. The quick freezing technique was applied on 4- and 8-cell (day-3) mouse embryos of hybrids. The in vitro development of frozen thawed 4- and 8-cell embryos (23% and 21% respectively) was found to be significantly lower than that of frozen thawed morulae (89%). Permeation in glycerol-solutions before equilibration significantly increased survival of 4- and 8-cell embryos (66% and 77% respectively). By the use of dimethylsulfoxid (DMSO) in the permeation solutions an even higher survival rate was obtained in the cryopreservation of 8-cell mouse embryos (95%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979856 TI - Xenografting of human pleural mesotheliomas to athymic rats and mice. AB - We xenografted cells from eight human pleural mesotheliomas to athymic rats and from one tumour to athymic mice. Four of the tumours grew for one or more passages in rats. Two of the tumours could be serially long-term passaged in athymic animals over a period of 33 months for 27 passages and 41 months for 18 passages, respectively. The growth rates of the two long-term transplantable tumours increased during the initial passages, after which the tumour-volume doubling time remained 3-5 days. Only one of the nine transplanted mesotheliomas exhibited growth characteristics suitable for drug-testing experiments. PMID- 2979857 TI - Bone marrow and peripheral blood modifications in C57BL mice administered with cyclophosphamide and levan. AB - The effects of cyclophosphamide (CY) and levan at low (0.1 mg) and high (10 mg) doses (previously found to inhibit and stimulate, respectively, B16 melanoma growth) were studied. in single and combined administration, on the blood and bone marrow of C57BL mice. Very marked changes in the cellular composition of blood and bone marrow were induced by the various treatments. CY caused a sharp depletion of leukocytes in blood and bone marrow followed by a restoration of granulocytes while lymphocytes remained low. Levan caused a sharp dose-dependent depletion in bone marrow lymphocytes. Blood lymphocytes were not affected, however, by the polysaccharide. With the high levan dose, after a temporary granulocytopenia and monocytopenia, prominent granulocytosis was observed. While CY affected equally the different types of cells, levan decreased the number of lymphocytes and erythroblasts to a greater degree than the number of granulocytes. In the combined CY-levan treatments, the low levan dose generally attenuated the suppressive effect of CY, while the high concentration aggravated it. PMID- 2979858 TI - Heterotransplantation of human malignant endometrial tumors into nude mice. AB - Seven of 13 human malignant endometrial tumors heterotransplanted into nude mice were accepted. The graft acceptance was significantly higher in papillary than in endometroid adenocarcinomas (p less than 0.01), whereas there was no observed relation between transplantability rate and other clinical prognostic factors. Of the carcinosarcomas and endometroid adenocarcinomas only PgR negative tumors were accepted. PMID- 2979859 TI - Prostate-derived growth factors for bone cells: implications for bone physiology and pathophysiology: review. AB - Metastatic prostatic cancer is virtually unique in its tendency to induce osteoblastic lesions. Several theories have been proposed for the explanation of this phenomenon, among them, the hypothesis of growth substances secreted locally by the metastatic prostatic cancer cells. Thus the present review summarizes today's stage of knowledge on bone cell physiology and presents the results of several studies which have tested the mitogenic capacity of extracts from prostatic cancer tissues on various isolated cell culture systems. PMID- 2979860 TI - High serum lactic acid concentration after laparotomy and peritonitis in iron stores depletion. An experimental study on the rat. AB - Total depletion of iron stores with a moderately decreased blood hemoglobin concentration (114 x 10 vs. 155 x 12 g/l in controls, n = 28) was produced in rats (n = 28) by rearing them for six weeks on an iron free diet, in iron free cages, starting at two weeks of age. The controls were treated at the same time in same conditions, except that they were given a normal supply of dietary iron. In all rats laparotomy and peritonitis by ligation and puncture of the caecum were carried out with ether anesthesia. Postoperative mortality and blood lactic acid concentration were followed for 120 minutes after the procedure. Postoperative mortality (8 of 28, 29% vs. 1 of 28, 4%) and blood lactic acid concentration (3.3 x 0.3 to 3.6 x 0.5 vs. 1.4 x 0.1 to 1.8 x 0.2 mmol/l) were higher (p less than 0.001) in the experimental group than in the control animals. The surviving rats (2.0 x 0.1 to 2.1 x 0.2 mmol/l) had a lower (p less than 0.001) blood lactic acid concentration than those who died (5.6 x 0.5 to 6.1 x 1.1). The results suggest that total lack of iron producing a moderate decrease of blood hemoglobin concentration creates a significant risk in abdominal surgery and focus the activation of the anaerobic energy yielding process. PMID- 2979861 TI - Effect of phenobarbital and 3-methylcholanthrene on biphenyl hydroxylations and fluidity of rat liver microsomal membranes. AB - The metabolism of biphenyl by rat liver microsomes after administration of phenobarbital and 3-methylcholanthrene was studied. Phenobarbital increased the activity of biphenyl-4-hydroxylase and 3- methylcholanthrene increased the activity of both biphenyl-4-hydroxylase and biphenyl-2-hydroxylase as compared to non-treated (control) rats. Phenobarbital increased the lipid fluidity while 3 methylcholanthrene increased the lipid rigidity of microsomal membranes labeled with 1,6-diphenyl-1,3,5-hexatriene (DPH), as indicated by the steady-state fluorescence anisotropy [(ro/r)-1]-1. Arrhenius plots of [ro/r)-1]-1 indicated that the lipid phase separation of the control membrane at 22.1 +/- 1.1 degrees was reduced in phenobarbital treated (14.5 +/- 0.8 degrees) and increased in 3 methylcholanthrene treated rats (32.7 +/- 2.2 degrees). Arrhenius plots of biphenyl-4-hydroxylase and biphenyl-2-hydroxylase activities exhibited a break point at 21.8 +/- 1.1 degrees and 32.1 +/- 2.1 degrees, respectively, suggesting differences in the interactions of the enzymes with their annular lipids. It is suggested that biphenyl-4-hydroxylase requires a liquid state of its lipid microenvironment to be fully active, while biphenyl-2-hydroxylase a gel state of its lipid microenvironment. These studies provide a basis for postulating that a "non-genomic" mechanism of phenobarbital and 3-methylcholanthrene induces cytochrome P-450 dependent monoxygenases. PMID- 2979862 TI - Chemotherapy and vitamin E in treatment of Nb rat prostate tumors. AB - Combination of Vitamin E in a single megadose of 20 units per 200 gram rat and chemotherapy was performed. The results revealed that Vitamin E treatment does indeed reduce Adriamycin toxicity and adds to tumor reduction and decrease metastasis. Vitamin E also enhances the efficacy of Cyclophosphamide, Adriamycin and Methotrexate. PMID- 2979863 TI - The detection of dysplasias in ulcerative colitis by a monoclonal antibody: preliminary report. AB - The reactivity of dysplasia and of adenocarcinoma tissues with the monoclonal antibody 52-2 was tested in 40 specimens from colitic and non-colitic patients. These included specimens from colitic patients having abnormal glands with nuclear stratification and hyperchromasia, surrounded by severe inflammation. While the normal colonic mucosa demonstrated a positive reaction only at the base of the crypts (germinal epithelium), dysplasia in colitic patients demonstrated reactivity in the full length of the dysplastic glands: at the apical aspect of dysplastic cells, at the glycocalix, in the entire cytoplasm and the level of the cell membrane (between two contiguous dysplastic cells and at the stromal aspect of the cells). A similar reaction was found in dysplastic glands from colorectal adenomas in non-colitic patients. Invasive adenocarcinomas in colitis and in non colitics showed strong positive reaction. In colitics, no reactivity was found in abnormal glands with nuclear stratification with hyperchromasia, surrounded by severe chronic inflammation. The possibility of differentiating dysplasias from reactive reparative cellular changes in patients with ulcerative colitis is discussed. PMID- 2979865 TI - Sequence determinants of symptoms in the genomes of plant viruses, viroids, and satellites. PMID- 2979864 TI - Compensatory adrenal growth in dexamethasone treated male and female hamsters. AB - The aim of the study was to investigate the unilateral adrenalectomy - induced compensatory adrenal growth in dexamethasone treated male and female hamsters (Mesocricetus auratus Waterhouse). Animals were treated for 5 days with 25 micrograms dexamethasone/animal/day or with 0.2 ml 0.9% NaCl. The first injection was made 0.5-1 h after monolateral adrenalectomy or sham operation. In three sets of experiments male and female hamsters received no injection. Only in one of three experiments was the relative adrenal weight of monoadrenalectomised male and female hamsters receiving no injections higher than in sham operated groups. In the case of NaCl treatment, only in one group of monoadrenalectomised males was the relative weight of the right adrenal higher than in the control group. On the contrary, in all three experiments with dexamethasone treatment the relative weight of the solitary adrenal was higher than in sham operated animals, while there was no difference among appropriate groups of females. In monoadrenalectomised dexamethasone-treated male hamsters, volume of the glomerulosa and reticularis zones was higher than in sham operated group. Neither the average cell volume in particular adrenocortical zones nor the number of parenchymal cells in the zones and in the entire cortex were changed due to adrenalectomy in dexamethasone treated males. There was no difference in all stereologic parameters studied when monoadrenalectomised and sham operated dexamethasone treated females were compared. Plasma cortisol level was lower in hemiadrenalectomised dexamethasone-administered males, while 3H-thymidine incorporation was higher in both male and female dexamethasone-treated hemiadrenalectomised hamsters. The obtained results demonstrate the evident sex dependent response of the hamster adrenal gland to monoadrenalectomy and dexamethasone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979866 TI - Morphological aspects of lithium toxicity. PMID- 2979867 TI - [The medicolegal aspects of phosphine poisoning]. PMID- 2979868 TI - Tricyclic antidepressants: clinical considerations, tissue concentrations and morphopathologic changes in the acute intoxication in man. PMID- 2979869 TI - Investigation of fatal bipyridyl poisonings. PMID- 2979870 TI - Legal medicine questions connected with toxicoses due to combustion cases. PMID- 2979871 TI - [Sudden death in epileptics treated with valproic acid]. PMID- 2979873 TI - A contribution to the application of spectral methods in examination of intoxications. PMID- 2979872 TI - [A case of poisoning by methylene chloride during the spraying of a new cargo vessel]. PMID- 2979874 TI - Methodology and interpretation of morphiate detection in urine specimens. PMID- 2979875 TI - Biochemical aspects of lithium toxicity. PMID- 2979876 TI - [Poisoning by alcohol, narcotics and psychotropic drugs. A study of alcohol-drug associations in forensic medical thanatological practice]. PMID- 2979877 TI - Quantitative determination of morphine in hair: a comparison between RIA and HPLC methods. PMID- 2979878 TI - Fatal central effects of diazepam potentiated by alcohol and haldol. PMID- 2979879 TI - Thin-layer and gas chromatography techniques for the isolation and identification of carbamate pesticides in post mortem material. PMID- 2979880 TI - The medico-legal investigation of narcotic related deaths in Belgrade: 1980-1984. PMID- 2979881 TI - [Familial thallium poisoning. A study of 4 cases]. PMID- 2979882 TI - Microcomputer-based data register in the forensic toxicological laboratory. PMID- 2979883 TI - Studies on paraquat poisoning. PMID- 2979884 TI - Investigating effects of alcohol and medicaments in traffic casualties. PMID- 2979886 TI - Some questions concerning the use of safety belts in road traffic. PMID- 2979885 TI - [The finding of clues for the reconstruction of cases of being run over by a train]. PMID- 2979887 TI - [The expert assessment of whiplash injury with reference to the accident sequence]. PMID- 2979888 TI - Blood alcohol levels in motor vehicle accident fatalities in the Republic of South Africa. PMID- 2979889 TI - Driving and alcohol. PMID- 2979890 TI - Roentgenography and tomography as techniques used in forensic medicine. PMID- 2979892 TI - Post mortem detection of tooth coloured dental restorations by ultra violet radiation. PMID- 2979891 TI - [The value and limits of using perikymata in legal medicine and epidemiology]. PMID- 2979893 TI - The use of a microcomputer system for worldwide dental charting comparison. PMID- 2979894 TI - [The importance of clinical and postmortem toxicological analyses and their legal relationships]. PMID- 2979895 TI - The contribution of dental histology to forensic medicine. PMID- 2979896 TI - Identification of unknown bodies from the point of view of forensic dentistry. PMID- 2979897 TI - Studies on sex determination from palate and canine and on palatal development by the moire topograph. PMID- 2979898 TI - [The determination of alcohol in the vitreous humor. Its correlation with blood alcohol]. PMID- 2979899 TI - [The measurement of biochemical parameters in the vitreous humor. Their relationships to the period of time passed since death]. PMID- 2979900 TI - Amitriptyline and its metabolites: blood and urine levels in overdose. PMID- 2979901 TI - Active metabolites of psychotropic drug implications in legal toxicology. PMID- 2979902 TI - Pharmacokinetic, toxicological and morphopathological considerations on some Rumanian psychotropic drugs. PMID- 2979903 TI - Trend of fatal poisonings during the past five years in the activity of the National Institute of Forensic Toxicology. PMID- 2979904 TI - [The assessment of the nontoxic methanol level in body fluids and tissues]. PMID- 2979905 TI - Energy transport at the toxic damaged heart muscle cells. PMID- 2979906 TI - [Psychotropic substances, the presence and demonstration of narcotics in Budapest]. PMID- 2979907 TI - Importance of alveolar pulmonary gas analysis in mass-disaster victims. PMID- 2979908 TI - A virus-inducible tobacco gene encoding a glycine-rich protein shares putative regulatory elements with the ribulose bisphosphate carboxylase small subunit gene. AB - cDNA to an mRNA that is strongly induced in Samsun NN tobacco after tobacco mosaic virus (TMV) infection or salicylic acid treatment was used to probe a genomic blot and to screen a genomic library. The mRNA corresponds to a family of approximately eight genes, four of which were cloned. The sequence of the genes and flanking DNA in two clones was determined. One gene was found to contain an intron of 555 bp; S1-nuclease mapping studies indicated that this gene is expressed. The other gene is interrupted by an intron of 1,954 bp and is probably not expressed after TMV infection. The genes encode a protein of 109 amino acids with a putative N-terminal signal peptide of 26 amino acids. The protein contains a high proportion of glycine (25%) and charged amino acids (29%), suggesting that it may be a cell wall component. A comparison of the upstream sequences of the genes encoding the glycine-rich protein and the pathogenesis-related protein 1a showed only limited homology, although both genes are TMV- and salicylic acid inducible. However, the upstream sequence of the glycine-rich protein gene contains a 64-bp inverted repeat that occurs in a similar position in the tobacco ribulose bisphosphate carboxylase small subunit gene. PMID- 2979909 TI - Identification and characterization of the nodD gene in Rhizobium leguminosarum strain 1001. AB - A gene library of the symbiotic 240-kb plasmid of Rhizobium leguminosarum strain 1001 was constructed in pUC18. The clones showing homology with a 6.6-kb fragment containing nodEFDABC from the Sym plasmid pRLlJI were detected by colony hybridization. Additional probes from the symbiotic region of pRLlJI were used to localize the corresponding genes on the map of pRle1001a. The relative positions of nod and nif gene clusters are different than those of pRLlJI. A comparison of the amino acid sequence for NodD from pRle1001a with NodD proteins from other Rhizobium species showed a high degree of sequence conservation at the amino terminus of the protein. PMID- 2979911 TI - Extended use of the internal mammary-coronary artery bypass. AB - Extended use of the IMA for routine coronary bypass grafting should do much to reduce the eight- to ten-year failure rate seen so frequently with the saphenous vein graft. However, considerable judgment is necessary for its proper use in a large number of situations and the performance of multiple sequential IMA anastomoses is technically more difficult than with saphenous veins. Special instruments have been devised to improve handling and suturing of the IMA and, as with the saphenous vein graft operation, performance of these anastomoses will be improved with increased experience. Extended use of the IMA is the operation of choice for younger patients with good left ventricular function and offers significant advantages to a group of patients that stands to benefit most by preventing both early and late conduit failure. PMID- 2979912 TI - The use of free-hand unstented aortic valve allografts for replacement of the aortic valve. PMID- 2979910 TI - The avirulence gene avrBs1 from Xanthomonas campestris pv. vesicatoria encodes a 50-kD protein. AB - A gene cloned from Xanthomonas campestris pv. vesicatoria race 2, avrBs1, specified avirulence on pepper cultivars containing the resistance gene Bs1. A series of exonuclease III deletions were made on a 3.2-kbp DNA fragment that determined full avirulence activity, observed as hypersensitive response (HR) induction. The deletion products were subcloned into the broad host range cloning vector pLAFR3, conjugated into a virulent X. c. pv. vesicatoria race 1 strain, 82 8, and scored for their ability to induce a HR on a pepper cultivar (ECW10R) containing the resistance gene Bs1. A span of approximately 1.8 kbp of DNA was necessary for full induction of the HR. The nucleotide sequence revealed two open reading frames (ORFs) capable of encoding proteins of 12.3 and 49.8 kD, designated ORF1 and ORF2, respectively. Deletions into ORF1 altered the HR inducing activity to give an intermediate phenotype. Deletions into ORF2 completely destroyed activity. When the ORF2 coding region was driven by the lacZ promoter on plasmid pLAFR3 (placD), full avirulence activity was restored, indicating that ORF2 alone can induce the HR. Antisera raised to a beta galactosidase-ORF2 fusion protein reacted with a 50-kD protein in X. c. pv. vesicatoria race 1 (placD) transconjugants. The deduced amino acid sequence of ORF2 had approximately 47% overall homology to the carboxyl terminus of the avirulence gene, avrA, isolated from Pseudomonas syringae pv. glycinea race 6, and 86% homology over a region of 49 amino acids. P. s. pv. glycinea, however, did not induce an HR on ECW10R plants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979913 TI - Artificial heart implantation for end-stage cardiac disease. PMID- 2979914 TI - Repair of ascending aortic aneurysms and dissections. PMID- 2979915 TI - The surgical approach to ventricular tachyarrhythmias. PMID- 2979916 TI - Operative technique and considerations influencing choice of operation to bypass the right ventricle. PMID- 2979917 TI - Surgical treatment of the Wolff-Parkinson-White syndrome and other supraventricular tachyarrhythmias. PMID- 2979918 TI - Repair of anterior mitral leaflet chordal rupture or elongation (the flip-over technique). PMID- 2979919 TI - Surgical reperfusion of acute myocardial ischemia: a clinical review. PMID- 2979920 TI - Arterial switch operation for transposition of the great arteries with intact ventricular septum. PMID- 2979921 TI - Mitral valve reconstruction for mitral regurgitation. PMID- 2979922 TI - Concomitant mitral valve repair or replacement and coronary revascularization. AB - Patients undergoing combined mitral valve replacement and coronary revascularization require surgical skill and especially judgment for optimal results. In our hands, cardioplegia has not been a pivotal event in affecting survival, and this probably relates to our previous philosophy of limiting the hypothermic ischemic episodes to 15-minute intervals. Currently, we believe that valve repair, when it can be accomplished, is preferable to valve replacement, especially in the patient with ischemic mitral valve disease. When repair cannot be satisfactorily accomplished, replacement with retention of the posterior leaflet seems clinically to be associated with less disturbance of left ventricular function. PMID- 2979923 TI - Technique and results of tricuspid annuloplasty. PMID- 2979924 TI - Surgical treatment of pulmonary atresia with intact ventricular septum. AB - A concept of management of pulmonary atresia in infancy has been presented. The basic theme employed is that pulmonary valvotomy or an outflow patch should be applied to most infants so that a biventricular heart is achieved in as many children as possible. Clearly, a functioning right ventricle cannot result in all instances and systemic to pulmonary shunts have an important adjunctive and sometimes primary therapeutic role. Our ability to judge right ventricular capability has been improved by preoperative assessment of segmental cavitary hypoplasia by noninvasive echocardiography and angiographic techniques. PMID- 2979925 TI - Repair of complete atrioventricular canal: 1975-1985. PMID- 2979927 TI - The IMA graft: current application and technique. PMID- 2979926 TI - Myocardial protection. AB - Early studies of myocardial protection were designed to minimize ischemic injury. The next class and generation of investigations will most likely be designed to accelerate recovery following known myocardial injury. Such techniques will play an important role in allowing operations on acutely injured and ischemic myocardium and will be important in the treatment of postischemic injury when such injury occurs during the course of complex cardiac operations. Surgical aspects of myocardial metabolism are still rudimentary and many empiric observations require further exploration into the mechanisms by which such applications work. PMID- 2979929 TI - Treatment of ascending aortic dissection in the Marfan syndrome. PMID- 2979928 TI - Surgical management of combined carotid and coronary occlusive disease. PMID- 2979930 TI - Techniques for management of the calcified mitral annulus. AB - Mitral annulus calcification is most often associated with myxomatous degeneration of the mitral valve. It is characterized by a horseshoe-shaped area of mitral annulus calcification that extends into the ventricular cavity to varying degrees. Standard suture techniques may be associated with fracture of the calcification and subsequent paravalvular leak. Seventeen patients aged 54 to 85 (mean 68.1) had mitral valve replacement using four techniques: suture through calcified annulus (3); decalcification (6); onlay patch (5); and drill technique (3). There were no hospital deaths, and one 81-year-old patient with associated coronary artery disease and ascending aortic calcification succumbed to a cerebrovascular accident three months postoperatively. The remaining patients are living and active with functional improvement. Follow-up has ranged from seven months to seven years with a mean of 4.0 years. One patient has hemodynamically insignificant mitral regurgitation after use of an onlay patch technique. No one technique could be singled out as clearly advantageous over the others from this series of patients. PMID- 2979931 TI - Eicosanoids (prostaglandins) and the cardiovascular system. PMID- 2979933 TI - Pulmonary artery approach to the aortic root in bacterial endocarditis. PMID- 2979932 TI - Aortic valve replacement: continuous-suture technique. PMID- 2979934 TI - The rationale for skeletal-muscle-powered counterpulsation devices: an overview. PMID- 2979935 TI - Reassessing right ventricular function and ventricular interaction: the role of global myocardial contractile mechanics. PMID- 2979936 TI - Trace metals in cement and in airborne cement dust. PMID- 2979937 TI - Proposed contingency plan and field management of hazardous spills in Egypt. PMID- 2979938 TI - Hepatotoxicity of some antimoulting compounds in white male mice. PMID- 2979939 TI - House flies (Musca domestica) as a vector of some enteric pathogens in an Egyptian village. PMID- 2979940 TI - Abnormalities in the sand fly Phlebotomus papatasi Scopoli (Diptera: Psychodidae) of Egypt. PMID- 2979942 TI - Risk factors and outcome of hospital-acquired acute renal failure (clinical epidemiologic study). PMID- 2979941 TI - Immunological detection of Plasmodium falciparum circumsporozoite antigens in Anopheles sergentii from Egypt. PMID- 2979943 TI - Orodental and ultrastructural studies of pulp and gingiva in phenylketonuria. PMID- 2979944 TI - Health education needed to improve public STD awareness in Somalia. PMID- 2979945 TI - Immunological changes in typhoid fever. PMID- 2979946 TI - An outbreak of meningococcal infection at the time of pilgrimage in Saudi Arabia. PMID- 2979947 TI - The protective effect of free radical inhibitors against catecholamine-induced toxicity. PMID- 2979948 TI - Bovine leptospirosis: bacteriologic versus serologic diagnosis of cows at slaughter. PMID- 2979949 TI - Effect of oral zinc supplementation on the cell mediated immunity in lepromatous leprosy. PMID- 2979950 TI - Observations on bionomics of mosquitoes in Ain-Ghosin village Ismailia Governorate. PMID- 2979951 TI - A study of the knowledge, attitude and practice of the Egyptian consumer towards the possible introduction of radionuclides contaminated foods to the market. PMID- 2979952 TI - Evaluation of the effectiveness of nutrition counseling programme in Kuwait. PMID- 2979953 TI - Effect of the administration of vitamin B6 at two levels of intake on xanthurenic acid excretion among oral contraceptive pill users. PMID- 2979954 TI - Determination of lethality (LD50) of common trace elements. PMID- 2979955 TI - Uptake and metabolism of estramustine in the Dunning R3327H tumour. AB - A time-dependent plasma, tumour cell and nuclear uptake was found after intraperitoneal 3H-estramustine administration, in rats bearing the Dunning H tumour. Uptake of the drug into tumour cells had not reached a maximum after 4 h, whereas no real change was seen in the nuclear fraction after 2 h. Binding to the nuclear protein matrix of Dunning H tumour cells of 3H-estramustine was also found (2.46 pmol/g protein). High performance liquid chromatography demonstrated the existence of estramustine, its oxidized metabolite estromustine, estradiol and estrone in the plasma and the tumour after intravenous administration of 100 microCi 3H-estramustine. PMID- 2979956 TI - Correction of thoracic aortic aneurysm-bronchial fistula. AB - The descending thoracic aorta is the most common site for thoracic aneurysms to occur and may present asymptomatically or manifest itself with an array of symptoms depending on the involvement of adjacent structures. Hemoptysis may occur due to erosion of the aneurysm into a bronchus or the pulmonary parenchyma. This report describes the technique in a patient with a descending thoracic aneurysm who underwent emergent resection of the aneurysm with en bloc removal of the involved left lower lobe of the lung. PMID- 2979957 TI - Clinical application of left ventricular assist devices. AB - In Japan, 32 patients have had application of monoventricular and biventricular assist devices during the past three years. Five of the 32 patients treated by the Fall of 1986 have successfully achieved long-term survival. In this paper we describe our experience with the Tomasu and Pierce VAD in a total of four and two patients, respectively. Four of the six patients could be successfully weaned from the VAD and two of them were long-term survivors. Nonsynchronizing pumping of the VADs was effective, as well as synchronizing pumping. Anticoagulant therapy is highly recommended during the use of the VAD although there was no significant incidence of thromboembolism or thrombus in the devices in this clinical series. PMID- 2979958 TI - Technical aspects of total artificial heart implantation for temporary applications. AB - Temporary use of the Jarvik-7 artificial heart in critically ill patients awaiting transplantation has prevented death in greater than 60% of attempts. Success is very dependent on meticulous surgical technique. Failures in the operating room cascade into enlarging problems postoperatively. Appropriately selected patients implanted without bleeding or fit problems recover quickly and are suitable candidates for transplantation within a few days. Even acute renal failure or pulmonary insufficiency from end-stage cardiac disease rapidly reverses upon restoration of normal hemodynamics. Anticoagulation is mandatory to prevent thromboembolism and continuous heparin plus oral dipyridamole is the most successful regimen to date. Primary intraoperative security of all suture lines or potential bleeding foci is thus essential. Bleeding has been associated with tamponade, pulmonary complications, and transfusion stimulated anti-HLA antibodies which can prevent subsequent transplantation. The length and suturing technique of the aortic conduit are critical. Compression of the mediastinal structures by the device can impede systemic or pulmonary venous return, with disastrous results. Displacement of the blood pumps away from the midline, usually into the pleural space, allows for more room when closing the sternum. The 100 cc blood pump fits best in patients greater than 85 kg, while the 70 cc model can be implanted in patients as small as 50-60 kg. PMID- 2979959 TI - Implantation of the left ventricular assist device. AB - Development of mechanical devices for support of the failing heart is a major goal in cardiac surgery. The application of left ventricular assist device (LVAD) is a promising approach in the case of severe and otherwise untreatable cardiac failure. In our experience we have used two external centrifugal pumps for the extracorporeal biventricular cardiac support in a post-transplantation patient who experienced severe rejection six months after heart transplantation. Our own series includes a total of ten implantations of LVAD's with six patients who could be weaned from the device but only one long-term survivor. The clinical results are not encouraging which suggests that the heart of the patient who needs an LVAD has been damaged beyond any chance for later recovery. Obviously timing is the most crucial aspect of the decision to implant the device. It would appear that orthotopic implantation of the transplanted heart remains the method of definitive treatment. PMID- 2979960 TI - Tetralogy of Fallot repair with minimal or no ventriculotomy. AB - Twenty-four consecutive patients with the tetralogy of Fallot underwent repair from March 25, 1985 to March 24, 1986. Correction was achieved without a ventriculotomy in 8 patients, necessitated a minimal ventriculotomy in 8, and a formal right ventriculotomy incision was required in the other 8. There were no early or late phase deaths-0% (70% C.L. 0%-7%). Post repair right ventricular to left ventricular pressure ratios were not significantly different between the three groups (p = 0.85). The 16 patients (66%) who underwent repair without a formal ventriculotomy were found to have severe hypertrophy of the infundibular septum as the primary etiology for right ventricular outflow tract obstruction. The other 8 patients (34%), who required a transventricular approach to the repair, were all found to have predominantly hypoplasia of the outflow tract and not hypertrophy. We conclude that repair of the tetralogy of Fallot can frequently be accomplished with no or a minimal ventriculotomy, and the results are satisfactory. Herein is described the operative approach to this procedure. PMID- 2979961 TI - Intraoperative angioplasty and coronary artery bypass grafting for the treatment of multisegmental coronary artery disease. AB - This report summarizes our experience with intraoperative angioplasty and coronary artery bypass graft surgery for the treatment of multisegmental coronary artery disease. The indication for intraoperative angioplasty was the presence of diffuse, symptomatic, coronary artery disease with at least one coronary artery having two or more areas of narrowing. The balloon catheter was introduced through an arteriotomy and advanced through the point of obstruction. The balloon was then inflated to 6 atmospheres for 15-20 seconds. This procedure was repeated two to three times. The time required for this technique averaged less than 5 minutes. Although there were no documented intraoperative perforations, intimal injuries were observed in two patients at the time of postoperative catheterization. Early and late postoperative angiograms showed that approximately 30% of angioplastied vessels had an improvement in their dilated lesions. Patients with discrete areas of narrowing had greater improvement in luminal diameter than those with diffuse narrowing (49% versus 17%). There was no difference in graft patency or left ventricular wall motion between vessels that did or did not undergo angioplasty. PMID- 2979962 TI - Coronary artery spasm. AB - Coronary artery spasm is a transient reduction in lumen diameter of an epicardial coronary artery of sufficient degree to produce objective evidence of myocardial ischemia in the absence of any significant increase in heart rate or blood pressure. In this article are summarized pathophysiological observations, the coronary angiographic anatomy of patients with coronary spasm, etiologic considerations, methods to provoke coronary artery artery spasm and their clinical usefulness, the role of coronary artery spasm in patients with clinical angina pectoris and myocardial infarction, and finally, the role of coronary artery spasm in patients undergoing coronary artery surgery. PMID- 2979963 TI - Cardiac valve prostheses: pathological and bioengineering considerations. AB - Cardiac valve replacement with mechanical prosthetic or bioprosthetic devices enhances patient survival and quality of life. Nevertheless, prosthesis associated complications are frequent and contribute significantly to outcome. Thromboembolic complications are the most important problems in patients with mechanical valves, necessitating chronic anticoagulation in all patients receiving them. In contrast, patients with bioprosthetic valves, composed of chemically treated animal tissues, generally do not require anticoagulants. However, bioprostheses fail frequently by degeneration, especially that involving cuspal calcification. This paper reviews the pathological and bioengineering considerations in the selection of cardiac prosthetic valves and the management of patients who have received these devices. The significance, morphology, and pathogenesis of the observed major complications and other alterations during function are described in detail. Contemporary investigative trends are summarized, including studies of inhibition of mineralization and other degenerative changes in bioprostheses, improved design rigid mechanical valves with pyrolytic carbon occluders and the development of central-flow, flexible polymeric leaflet valves. PMID- 2979965 TI - Replacement of the aortic valve with cryopreserved aortic valve allograft: considerations and techniques in children. AB - Normal structure and function of the aortic valve requires not only the basic three-cusp architecture but also the dynamic action and shape of the aortic sinuses and correct diameter relationships at the left ventricular outlet and at the aortic sinus rim. Congenital malformation of the aortic valve implies abnormality of the aortic cusps and, additionally, there may be deformity of the aortic sinuses or the subvalvular left ventricular outflow tract or the supravalvular aorta. Aortic valve replacement in children using an aortic valve allograft often requires enlargement of the left ventricular outflow tract or restoration of aortic sinus configuration. Careful consideration of the malformations of the aortic root and planning of reconstructive operation is required lest the malformation be recreated in the allograft valve replacement. Even the best execution of known aortic root reconstructive or left ventricular outflow tract enlarging operations has limitations in restoration of normal structure and compliance of the aortic root. Consequently, there remain many unknown factors that could affect late results of aortic valve allograft operations in children compared to those results observed in adult patients. PMID- 2979964 TI - Extended aortic root replacement for treatment of left ventricular outflow tract obstruction. AB - Recurrent tunnel stenosis of the left ventricular outflow tract following operation for subaortic stenosis and hypoplastic aortic annulus remain a challenge for pediatric cardiac surgeons. We have recently applied a new technique of extended aortic root replacement using an aortic allograft to treat three patients who had previously been operated upon for subaortic stenosis and three who had aortic stenosis with a hypoplastic aortic annulus. This new procedure combines the concept of aortoventriculoplasty with allograft aortic root replacement and coronary artery reimplantation. The valved aortic homograft is used in place of an aortic valve prosthesis and the attached anterior mitral leaflet augments the interventricular septum to relieve the subvalvular left ventricular outflow tract obstruction. The coronary ostia are then reimplanted into the allograft and distal graft to ascending aorta anastomosis completed. Allograft aortic tissue is then used to patch the right ventricular outflow tract. There have been no operative or late deaths. One patient developed Serratia marcescens mediastinitis but recovered uneventfully after mediastinal drainage. Two cases of transient complete heart block reversed spontaneously. A patient with type II hyperlipidemia developed postpericardiotomy syndrome early, which resolved but then required reoperation at six months for stenosis of the distal anastomosis and left main coronary stenosis, both thought to be complications of his underlying disease. Completely benign convalescence and early follow-up has occurred in the last two patients. This modified technique using aortic allograft was very helpful in treating these difficult problems, and the lack of mortality, limited morbidity, and good function results are encouraging. PMID- 2979966 TI - Basic principles of cryobiology. AB - The ultimate aim of most cryopreservation procedures is the retention of the structural and functional integrity of the frozen cells or tissue. Designing strategies for achieving high "viability" of the frozen tissue requires an appreciation of the complex physical-chemical events which occur during freezing and thawing, as well as the importance of other processing steps including tissue procurement and post-thaw processing of the tissue, all of which can adversely affect the tissue. Here we have provided a general overview of cryobiology with special emphasis on techniques for organ preservation. PMID- 2979967 TI - Measurement of postcryopreservation viability. AB - For any tissue, there is a cell viability threshold below which the ability of the tissue to maintain itself and function will eventually be compromised. During cryopreservation and subsequent thawing of tissues there are many steps involved, each with attendant potential risks for reduction of viability. To determine the effectiveness of any cryopreservation procedure it is important to select appropriate assays. In this manuscript viability assays, in general, are reviewed from a biological viewpoint prior to review of methods employed for assessment of heart valve viability. Both in situ and in vitro assays of heart valve viability indicate that valve mechanical properties and the majority of fibroblasts, which are responsible for maintenance of the valve connective tissue, are retained after cryopreservation. PMID- 2979968 TI - The viable cryopreserved allograft aortic valve. AB - One hundred and twenty-four patients underwent aortic valve replacement with a nonviable 4 degrees C refrigerated aortic allograft valve. One hundred and eighty four patients underwent aortic valve replacement with a viable cryopreserved aortic allograft valve in a later era. The longest follow-up was 16 years for the group with the nonviable valve and 11 years for the group with the viable valves. Within this time frame, reoperation was required in 23 patients with nonviable valves for leaflet perforation or rupture whereas no patients in the group with viable valves developed this complication (p less than 0.0001). The prevalence of endocarditis and thromboembolism was very low in both groups. Viability of leaflet tissue is associated with an important improvement in durability over nonviable allograft valves. Consequently, long-term follow-up results of allograft valves might be best expressed in terms of viability. The current evidence suggests that the viable cells are donor in origin. The viable cryopreserved aortic allograft valve offers significant advantages over current nonviable allograft valves, mechanical valves, and bioprostheses. PMID- 2979969 TI - Application of homografts in clinical surgery. AB - Aortic freehand homografts have been used since 1962 and are still our preferred method of treatment. They are centrally flowing and are consequently nonobstructive and give rise to neither emboli nor turbulence. They do, however, undergo a slow process of degeneration, giving plenty of time for an elective low risk second operation. An additional advantage is that they can be used in children. Various methods of valve preparation and storage have been used, but there is no clear evidence that one particular method is superior. The position of the homograft in the heart does have a bearing on its function, with the valve persisting longer in the low-pressure right heart chambers. Long-term results indicate that the unmounted homograft in the aortic area gives better results than currently used bioprostheses and in the right ventricular outflow tract there is no comparable valve. Studies of the viability of the various valve components and also their immunological significance should help to improve the long term durability of the homograft valve. PMID- 2979971 TI - A modified free-hand technique for homograft aortic valve replacement. AB - A modified technique for trimming and inserting the aortic homograft valve is described. This altered free-hand technique is designed to maintain normal spacial geometry of the valve apparatus in order to reduce the risk of early homograft valve incompetence. PMID- 2979970 TI - "Fresh" aortic allografts: long-term results with free-hand aortic valve replacement. AB - Renewed interest in the use of "fresh" and cryopreserved allograft valves for aortic valve replacement (AVR) prompted an updated analysis of the long-term results of our old experience (1964-1971) with free-hand AVR. Eighty-three patients received "fresh" (antibiotic stored at 4 degrees C for intervals between 24 hrs and 18 days), free-hand allograft valves. Current (1986) follow-up was 96% complete; cumulative follow-up included 773 patient-years (pt-yr) and averaged 9 yrs. Importantly, 37 patients were still at risk with their original allograft valve at ten yrs, and 12 patients at 17 yrs. Standard conservative criteria were used to assess valve-related complications. Thromboembolism (TE) occurred at a linearized incidence of 1.0%/pt-yr, anticoagulation-related hemorrhage (ACH) at 0.2%/pt-yr), and fatal prosthetic valve endocarditis (PVE) at 0.5%/pt-yr. In actuarial terms, the incidence of degenerative valve failure was 30 +/- 6% (+/- SEM) at ten yrs and 40 +/- 7% at 15 yrs. Valve failure due to all causes (including sudden, unexplained deaths and PVE) occurred in 38 +/- 6% of patients at ten yrs and 57 +/- 7% after 15 yrs. The incidence of fatal valve failure was 11 +/- 4% at six yrs (the time of the last event). The rate of reoperation was 33 +/- 6% at ten yrs and 52 +/- 7% at 15 yrs. Given the relatively crude methods of allograft valve preparation and storage during this remote era, we believe that these long-term results with free-hand allograft AVR are satisfactory, albeit far from optimal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2979972 TI - The future of homografts. AB - The cryopreserved aortic valve is at least as satisfactory for aortic valve replacement as any known device. The future lies in fine-tuning the preservation process and sterilization. Concerning the use of allograft aortic valves as conduits, the allograft valved ascending aorta is the best conduit for the connection of right ventricle (or left ventricle) to pulmonary arteries. Availability, tailoring to size, and addition or need for composite parts remain to be developed. The allograft aorta and aortic valve may have great usefulness for replacing the aortic valve and ascending aorta in cases of aneurysms, infections, and congenitally narrowed left ventricular outflow tract; their use in these cases is yet to be increased. The future of allograft saphenous vein will involve in-vitro studies on cryopreservation toxicity, antibiotic toxicity, biochemical function of endothelial cells, and endothelial viability. Considerable in-vitro testing is necessary in both animal and human preparations. Also considered is the future of allograft small-caliber arteries--i.e., will they behave in a similar fashion to the intact internal mammary artery?--and the heart and peripheral vascular systems--i.e., are there any more spare parts? PMID- 2979973 TI - Transplantation of aortic and pulmonary allografts, enhanced viability of endothelial cells by cryopreservation, importance of histocompatibility. AB - Fresh heart valve allografts were preserved at 4 degrees C for 14 days, cryopreserved and stored for 63 days, and studied for endothelial viability and antigenicity, in order to obtain some information on the immunobiological status of allografts before transplantation. The surgical technique described by Ross for subcoronary position is preferred and briefly outlined. Four explanted incompatible allografts were studied by light and scanning electron microscopy and immunohistochemistry to assess the immunological reactions and tissue changes that occurred between 9 days and 16 weeks postoperatively. Valve leaflet motion and opening velocities were studied by echocardiography postoperatively to establish a baseline with which to distinguish early leaflet degeneration. Distensibility of the aortic annulus was studied postoperatively by supraaortic angiography to justify one of the goals of reconstruction of the aortic root with allografts. Antibiotic preserved allografts at 4 degrees C showed no viable endothelial cells after 8 days while the cryopreserved allografts demonstrated a high rate of viable endothelial cells capable of expressing surface antigens (HLA class I and II). Although the valve explants showed focal mononuclear cell infiltrations with T-lymphocytes, the allografts healed in place. The "classic" findings of rejection could not therefore be demonstrated. In summary, cryopreserved valve allografts, like the fresh, are antigenic. It is therefore recommended to use compatible valve grafts, when possible, which might be a positive step to improve the functional longevity of valve allografts. Immune response after valve allograft transplantation does not cause acute valvular dysfunction but rather chronic tissue changes which might lead to early degeneration of the allograft. The opening velocities of preserved aortic and pulmonary allografts were normal at 3-4 years postoperatively irrespective of histocompatibility. Echocardiography might be a useful tool to detect early degenerative changes of incompatible valve leaflets. The aortic root is distensible after allograft transplantation. PMID- 2979974 TI - Intraventricular tunnel repair of double outlet right ventricle. AB - Double outlet right ventricle (DORV) may be divided into subsets according to the position and commitment of the ventricular septal defect (VSD) to the great arteries. In DORV with subaortic VSD, an intraventricular tunnel repair is the recommended operation. The current hospital mortality is 5% with an actuarial survival of 83% at 15 years. DORV with doubly committed VSD should also be repaired with an intraventricular tunnel, and the surgical results are similar to those for DORV and subaortic VSD. In DORV with subpulmonary VSD, an intraventricular tunnel repair is advisable when the distance from the tricuspid to the pulmonary valve exceeds the distance from tricuspid to aortic valve. Otherwise, a spiral intraventricular tunnel or an arterial switch procedure should be considered. In DORV with doubly committed VSD, the results of surgical treatment have been less good, and alternative forms of surgical treatment require further evaluation. The surgical treatment of DORV with pulmonary stenosis and the surgical details of the intraventricular tunnel repair are discussed. PMID- 2979975 TI - Surgical repair of hemitruncus: principles and techniques. AB - Ten patients have presented at the World Children's Hospital of Melbourne with hemitruncus, a form of truncus arteriosus. Nine of these patients underwent successful one- or two-stage hemodynamic repair. Early repair is carried out to avoid pulmonary vascular disease of the lung directly supplied from the trunk and to recruit as much of the contralateral pulmonary artery vasculature bed as possible is a secondary goal. The repair should be contemplated within the first six months of life to prevent severe pulmonary vasculature obstructive disease. Anatomy that is repairable in one stage is when the left pulmonary artery originates from the trunk and the complete right pulmonary artery supplied by pulmonary collaterals from the arch to the descending aorta. Anatomy requiring two-stage repair occurs when the RPA rises from the trunk and the left PDA rises from the descending aorta or distal aortic notch and either the right or the left lung blood supply is from multiple origins and requires unifocalization. With these anatomical principles and the general meticulous perioperative and postoperative care of the infants, a high degree of success should be obtainable with repair of this hemitruncus. PMID- 2979976 TI - Surgical techniques for treatment of bacterial endocarditis of the mitral valve. AB - Bacterial endocarditis of the mitral valve appears to be much less common than bacterial endocarditis of the aortic valve. One of the main etiologic factors is the presence of degenerative lesions of the mitral apparatus, ballooning or mitral floppy valve. The surgical anatomy of the lesions is described: vegetations, perforations, rupture of chordae tendinae, abscess of the mitral ring observed in the isolated mitral endocarditis, mitral-aortic dislocation, abscesses and aneurysms of the mitral-aortic fibrosa and jet lesions on the anterior mitral leaflet. In the isolated primitive mitral infective lesions, all the technical skills are directed toward the prevention of the perivalvular leakage of the prostheses. Special procedures are described for the management of the abscesses of the mitral ring. In patients with mitral-aortic lesions, the main problem is treatment of the dislocation of the annuli or aneurysms of the mitral-aortic fibrosa. Despite technical advances, the surgical prognosis of the mitral endocarditis remains severe. In a personal series, the authors recorded a mortality of 12% in isolated mitral cases and 42% in the combined mitral-aortic patients. Early surgical treatment remains the most significant factor in decreasing the fatality of such lesions. PMID- 2979977 TI - The total artificial heart: indications and preliminary results. AB - The development of the total artificial heart (TAH) has reached a level where it is now available for clinical applications. The TAH has demonstrated distinct advantages over other forms of mechanical circulatory assistance. As of December 1, 50 TAHs have been implanted: 5 as permanent devices, and 45 as a temporary mechanical bridge to cardiac transplantation. The use of the TAH has increased in the last several months, leading to a growing interest in defining the indications and contraindications to its use. End-stage cardiomyopathy (either idiopathic, ischemic, viral, or postpartum) has been the underlying disease in 80% of the TAH procedures to date. The TAH has also been applied in 5 cases of acute cardiac graft rejection, 2 cases of congenital heart diseases, and in one case after acute myocardial infarction. The indications for the use of the TAH in these and other potential patient groups is discussed in light of the current clinical results. PMID- 2979978 TI - Heterotopic cardiac transplantation: current status. AB - Heterotopic cardiac transplantation first introduced by Bernard in 1974 currently is rarely used as the procedure of choice when orthotopic cardiac transplantation can be considered. Specific indications for heterotopic cardiac transplantation include elevation of pulmonary vascular resistance and availability of a small or poorly functioning donor organ for a mortally ill recipient. Most cardiac transplant centers have abandoned its routine use because the recipient's diseased and poorly functioning heart remains as a potential source for embolism, infection, and continued angina, because the operative procedure is more complicated. Pulmonary complications are common due to compressive atelectasis of the right lung. Experience indicates that the heterotopic procedure is useful for those selected individuals in whom the orthotopic procedure is not appropriate and that rates of survival are nearly equal. PMID- 2979979 TI - Coronary reperfusion following experimental myocardial infarction. AB - Numerous studies have shown that early coronary reperfusion is feasible in the setting of evolving acute myocardial infarction in man. While early reperfusion reduces myocardial infarct size, there are potentially deleterious consequences of reperfusion. The concept of "reperfusion injury", oxygen-free radical damage, no reflow phenomenon, and stunned myocardium are discussed. PMID- 2979980 TI - Calcium channel blockers and cardiac surgery. AB - Calcium channel blockers have an important role in the pharmacotherapy of cardiovascular disorders. These agents act by inhibiting the slow inward current into excitable cells, exert direct negative inotropic, chronotropic, and dromotropic activity, and are potent vasodilators. These direct effects are modified by reflex autonomic stimulation and by pathologic states. Serious adverse effects of the calcium channel blockers are most frequently observed in patients with ventricular dysfunction, conduction system disease, or concomitant beta blockade. Calcium channel blockers are indicated in the treatment of angina pectoris, supraventricular arrhythmias, and hypertension. The use of these agents in patients with hypertrophic cardiomyopathy, congestive heart failure, and pulmonary hypertension is investigational. The calcium channel blockers are gaining increased importance in the management of patients undergoing cardiac surgery. Verapamil is indicated for the treatment of post-cardiac-surgical atrial flutter and fibrillation; however, the calcium antagonists are not effective as prophylaxis against postoperative supraventricular arrhythmias. Laboratory studies have shown that drug interactions exist between calcium channel blockers and inhalational anesthetics and nondepolarizing neuromuscular blocking agents; clinical studies have demonstrated that these interactions are rarely significant. Perioperative coronary spasm can be effectively treated with the calcium channel blockers. The timing of calcium antagonist withdrawal prior to surgery is controversial, but continuation of therapy until surgery is usually safe. The clinical significance of platelet function inhibition by the calcium antagonists is unknown. Protection of ischemic myocardium by calcium channel blockers has been demonstrated. Important interactions between the calcium antagonists, hypothermia, and the ionic constituents of cardioplegia require further study before the role of these agents as adjuncts to clinical cardioplegia is defined. Expanded indications and the introduction of new calcium channel blockers will result in increased use of these agents in the future. PMID- 2979981 TI - Closed-chest decannulation of transthoracically inserted aortic balloon catheter without grafting. AB - A method is presented which allows removal of a balloon assist catheter inserted directly (without a graft) through the ascending aorta in the course of heart surgery without the need for reopening the sternotomy incision. The catheter is inserted through the aortic wall under the protection of two purse string sutures which are temporarily tightened using implantable grade silastic rubber tourniquet. The end of the tourniquet is placed subcutaneously in a subxiphoid position. At the time of discontinuation of balloon assist, the balloon can be removed using local anesthesia without reopening the sternum by exposing the end of the tourniquet substernally, removing the catheter, and plugging the tourniquet. The silastic tourniquet may be left in indefinitely or removed through a similar exposure six to eight weeks after the procedure. PMID- 2979982 TI - Reoperation in patients with patent coronary bypass grafts. AB - As the population of patients having undergone myocardial revascularization increases, the number of reoperations will escalate and reoperations of patients with patent grafts will become increasingly frequent. Successful management of this new surgical entity requires a clear understanding of the probability of atherosclerotic involvement of patent vein grafts and careful attention to technical detail. Patent vein grafts more than five years old should be replaced to avoid the potential for embolization of atheromatous material and late vein graft closure. PMID- 2979983 TI - Left thoracotomy and femoro-femoral bypass for reoperative revascularization of the posterior coronary circulation. AB - Left thoracotomy and femoro-femoral cardiopulmonary bypass has been used for reoperation in five patients requiring coronary bypass graft into the inferolateral surface of the heart. Five patients had refractory angina pectoris and angiographic occlusion of lateral wall native vessels or previous occluded vein grafts and all had positive exercise test. Four of the five had patent internal mammary to the LAD. Following supine positioning and removal of the saphenous vein and isolation of the femoral artery and femoral vein, a left fifth interspace thoracotomy was made, the patient heparinized and cannulated for cardiopulmonary bypass, the pericardium opened, the heart dissected free, and either the internal mammary artery dissected off the left chest wall or saphenous vein grafts used to bypass the appropriate lesions. The proximal inflow was the descending thoracic aorta making tunnels for the vein grafts through the posterior pericardium. All of the patients did well in the postoperative period. This technique is recommended for reoperations in patients with documented inferolateral ischemia as the primary cause of symptomatology with mitigating circumstances against an anterior approach. PMID- 2979984 TI - Surgical management of difficult pacing problems in patients with congenital heart disease. AB - The child with congenital heart disease requiring permanent pacing presents a unique challenge with regard to the decision to pace, hardware, route, and unusual problems. Considerations of patient size, anatomy, insertion during and after complex intracardiac procedures, location of hardware, and unusual approaches are discussed including: placement of endocardial leads at open operation, closed transatrial endocardial technique, periprosthetic valvular endocardial placement, trans left superior vena cava placement and retromammary position via the axillary approach. Consideration of these unusual techniques may avoid frustration due to the complexities of the placement of pacing systems in these young patients. PMID- 2979985 TI - Excisional surgery for patients with ventricular tachyarrhythmias. AB - Thirty-eight patients with drug refractory ventricular tachycardia have been operated on since 1980. An attempt at preoperative localization of the site of arrhythmia was made in those patients who did not have severe ischemia or multifocal tachycardia. Operation consisted of intraoperative mapping, when feasible, subendocardial resection and cryolesions placed when sites of interest could not be resected. Patients averaged 56 +/- 44 years of age and 25 had associated coronary bypass. The operative mortality was 5/38 (13%) and two deaths were related to recurrent arrhythmia. Of 33 survivors, 21 (64%) were cured of their arrhythmia and an additional nine patients who had been drug refractory were controlled with medication alone. The best results were found in patients with anteroapical scar and unifocal tachycardia. Thus, direct operation for drug refractory ventricular tachycardia can be performed at a reasonable risk and with a high likelihood of successful arrhythmia control. PMID- 2979986 TI - Surgical treatment of partial anomalous pulmonary venous connection of the left lung (It-PAPVC). AB - Between January 1981 and December 1983, 4 patients, aged 6 to 50 years, with It PAPVC were operated upon at the Kokura Memorial Hospital, by a new technique of anastomosis between the left anomalous PV and the left auricular appendage. One patient, a 9-year-old male, had unilateral anomalous pulmonary venous connection of the entire left lung to the innominate vein. The other three patients had partial anomalous pulmonary venous connection from the left upper lobe to the innominate vein. Two patients has pulmonary valvular stenosis, and the other two had ASD. All patients were operated upon through a midsternal incision. After cardiopulmonary bypass was established, the anomalous vein was divided as high as was reachable. The present operative method involved the use of an oval pericardial gusset extending from the left auricular appendage into the split anomalous vein so as to obtain a wide anastomotic orifice. Other associated congenital anomalies were simultaneously corrected. All patients had uneventful recovery and at postoperative cardiac catheterization and angiography, the anastomosis between the pulmonary vein and the left auricular appendage was widely patent. PMID- 2979987 TI - Mechanisms of myocardial cell injury during ischemia and reperfusion. AB - Ischemia in myocardial tissue results in impaired high energy phosphate production and diminished perfusion of the interstitial space. Reduction in the supply of ATP to the sarcolemmal and sarcoplasmic reticulum Na+ and Ca2+ pumps results in a rise in intracellular (Ca2+), which can exceed normal systolic levels within 10 to 15 minutes. Elevated (Ca2+)i can cause activation of proteases and phospholipases, which can damage the sarcolemma and release toxic metabolites, such as lysophospholipids. Oxygen free radicals can be produced by breakdown of nucleosides and accumulate in the interstitial space. Accumulation of metabolites intracellularly can cause cell swelling, which in addition to rigor due to ATP depletion, can stress the weakened sarcolemma, producing cell rupture and death. With reperfusion, additional injury to the myocyte may occur. Resupply of oxygen can result in a burst of oxygen free radical production. Resynthesis of ATP may sensitize the myofilaments to Ca2+, resulting in a hypercontracture that can further promote cell rupture. Resupply of ATP and washout of H+ may cause activation of Na/Ca2+ exchange, thus intensifying Ca2+ overload. Washout of the hypertonic interstitial space fluid may aggravate cell swelling and produce sarcolemmal rupture. Prevention or alteration of ischemic and reperfusion injury in myocardial cells is important clinically. Strategies currently being explored include reducing the rise in (Ca2+)i, which occurs during ischemia and reperfusion; inhibiting the actions of phospholipase on the cell membrane; decreasing free radical effects; and reducing stress on the sarcolemmal by prevention of cell swelling and hypercontracture. PMID- 2979988 TI - The rational selection of inotropic drugs in cardiac surgery. AB - Despite improvements in surgical technique and intraoperative myocardial protection, certain patients have need for inotropic drug support after cardiac surgery. This review examines drugs that are currently in use for inotropic support of the heart, including calcium, epinephrine, dopamine, dobutamine, isoproterenol, and amrinone. Patient factors that may have an impact on the selection of appropriate drugs are also examined. Application of these data to specific patients must be guided by the particular hemodynamic derangements present. Careful analysis of the specific hemodynamic disorder and tailoring of inotropic therapy to these abnormalities are crucial. Such a rational approach to the selection of inotropic agents requires continuous hemodynamic assessment and recognition that the patient's condition and needs may change rapidly early after heart surgery dictating adjustment of subsequent therapy. PMID- 2979989 TI - Failure of intraaortic balloon counterpulsation caused by pacing or other electrical artifacts: a new method of correction. AB - A method using an interface device is presented designed to correct malfunction of counterpulsation caused by pacing signals or other electronic interference in patients undergoing simultaneous pacing and counterpulsation. PMID- 2979990 TI - The Senning procedure for repair of d-transposition of the great arteries. AB - We report our results with the Senning procedure for repair of d-transposition of the great arteries (d-TGA) including techniques that we feel facilitate the operation. When the atrial septal flap is deficient at the site of an atrial septal defect, the flap is filleted to enlarge it using living tissue. This also has the effect of thinning the septal flap which enlarges the left pulmonary vein pathway located posterior to the floor of the new caval tunnels. Another technique includes the use of the upper end of the sulcus terminalis in the construction of the roof of the new caval tunnels. The upper posterior rim of the right atriotomy is left external to this suture line and can be used in construction of the new pulmonary venous pathway. The latter suture line is inferior to the SA node and can be placed expeditiously and with less chance of damaging the SA node. Use of this technique has resulted in a reduced incidence of late arrhythmias. Following the Senning procedure in 40 patients there were no hospital deaths, but there were two late deaths. The Senning operation is currently a very satisfactory alternative for repair of d-TGA when arterial switch repair cannot be performed. PMID- 2979991 TI - Pulmonary autograft: an American experience. AB - Pulmonary valve autograft replacement of the aortic valve has been performed in nine patients in an effort to duplicate in the United States the experience of Sir Donald Ross in London. Four male and five female patients ranging in age from seven to thirty-four years have been operated on for indications of aortic stenosis in six and aortic regurgitation in three. Three patients had associated procedures performed concomitantly. All patients survived the surgery and were clinically improved, however, one patient did require reoperation for progressive aortic insufficiency. Specific indications, results, and technical considerations are discussed and illustrated. PMID- 2979992 TI - The timing of operation in valvular insufficiency. AB - The timing of operation is discussed for aortic, mitral, and combined aortic and mitral insufficiency. The effect of surgical mortality on selection is assessed on the basis of the Green Lane Hospital results for valve replacement in the current cardioplegic era. Particular attention is paid to the effect of the preoperative symptomatic status. The criteria for selection of the patient who is essentially asymptomatic are documented in detail. It is concluded that they differ, depending upon whether there is aortic incompetence, mitral incompetence, or incompetence of both valves. The effect of the type of valve used for replacement on case selection includes an up-to-date assessment of the results achieved with freehand aortic homograft valve replacement. PMID- 2979993 TI - Digitalis toxicity: mechanisms, diagnosis, and management. AB - Digitalis glycosides have a narrow margin between therapeutic and toxic levels. Although the incidence of digitalis toxicity appears to be decreasing, continuation of digoxin therapy in the face of overt toxicity carries a substantial risk of life-threatening cardiac rhythm disturbances. This review will focus primarily on toxicity produced by digoxin and discuss the mechanisms, clinical manifestations, and current management of digitalis toxicity. The appropriate methodology for measurement and interpretation of serum digoxin levels is emphasized. The varied cardiac dysrhythmias induced by digoxin are detailed and their specific management outlined. Immunological (antibody) treatment has now been established and approved for the therapy of advanced, life threatening digitalis toxicity and the multicenter trial of digitalis antibodies is described. Proper understanding of the pharmacokinetics of digitalis glycosides and careful followup of digitalis treated patients for early evidence of toxicity should help to decrease further the prevalence of toxic reactions. PMID- 2979994 TI - Coronary sinus interventions: from concept to clinics. AB - For almost 100 years the coronary venous system has attracted researchers as an access route to deprived myocardium. Different concepts have been tried and numerous experimental studies have been performed to evaluate whether coronary sinus occlusion, retroinfusion, and retroperfusion of arterial blood via the coronary sinus represent an effective treatment of myocardial ischemia. The early successful studies of Drs. Gregg, Eckstein, Beck and others led to the application of the Beck II procedure, a permanent retroperfusion technique used in the early 1950s to revascularize patients with diffuse atherosclerosis. The coronary sinus route was also used to retroperfuse blood intraoperatively during opening of the heart to keep the heart beating. Excessive mortality and severe side effects, such as myocardial edema and hemorrhage, and an insufficiently advanced technology resulted in the temporary demise of the coronary sinus approaches. Recently, however, the need to improve myocardial protection, despite enormous advances in coronary bypass surgery and interventional cardiology, has resulted in renewed interest in the coronary sinus as an access route to deprived myocardium. Advances in technology such as percutaneous catheter techniques have improved access to the coronary venous system and allowed for a physiological adaptation of coronary sinus retroperfusion techniques. Today the three major coronary sinus techniques, i.e., sychronized retroperfusion (SRP), retroinfusion of cardioplegia during cardiac arrest and retroinfusion of pharmaceutical agents in the normal working heart, and pressure-controlled intermittent coronary sinus occlusion (PICSO) have been documented as providing superior protection of jeopardized myocardium in selected subsets of patients. All of these techniques currently are under clinical consideration; the retroinfusion of cardioplegia has already found wide clinical acceptance, SRP and PICSO have only recently been tested in first clinical trials. Due to a vast resurgence in interest, it is desirable to survey the results obtained with each of these techniques, to discuss the pathophysiology and mode of action of coronary sinus interventions, and to assign them a place in the perspective of conventional therapies. Furthermore, an attempt will be made to weigh individual coronary sinus techniques against each other, discuss in which clinical settings each of them may be most effective, and define issues facing current development. PMID- 2979995 TI - A technique for selective graft perfusion during aortocoronary bypass. AB - A technique is described that provides early reperfusion of acutely ischemic myocardium during the construction of saphenous vein bypass grafts to the coronary arteries. Selective vein graft perfusion provides early and appropriate cooling of the most ischemic zones identified by either electrocardiographic changes or intramyocardial temperature probes and allows warm blood reperfusion during construction of proximal anastomoses. The technique may be applicable to patients who present with acute ischemia and evolving infarction, failed angioplasty, or left main coronary artery stenosis. PMID- 2979996 TI - The inframammary incision for median sternotomy in pediatrics. AB - Median sternotomy offers the ideal exposure for most cardiac and mediastinal procedures. One infrequently mentioned morbidity of the traditional median sternotomy is the unsightly scar, especially in pediatric and adolescent patients who are extremely sensitive about their body appearance. We report our experience using a bilateral inframammary "butterfly" incision on 31 patients in association with a median sternotomy to provide excellent exposure and an improved cosmetic result. PMID- 2979997 TI - Clinical experience with the Jahnke-Barron heart support. AB - The Jahnke-Barron heart support has proven to be a useful adjunct to coronary artery surgery by allowing an easy access to the coronary arteries while maintaining a quiet operating field. Further, the use of this device eliminates the need for a surgical assistant or a heart holder. PMID- 2979998 TI - Implantation of the automatic implantable cardioverter defibrillator. AB - Since February 1980 the automatic implantable cardioverter defibrillator has been implanted in over 1,500 patients. Sudden death rates have been reduced to 2%-4% annually. This report reviews the implantation techniques, their indications, and our clinical experience in 200 patients. PMID- 2979999 TI - Considerations in the surgical management of left atrial myxoma. AB - Myxomas are the most common tumors of the heart. They are frequently located in the left atrium and produce symptoms when they fragment and cause systemic emboli or when they interfere with cardiac valvular function and cause pulmonary congestion. Careful surgical management of these lesions should be curative with minimum early and late morbidity and mortality. Optimal operative technique emphasizes minimal manipulation of the heart before institution of cardiopulmonary bypass and aortic cross-clamping and careful examination of intracardiac chambers with meticulous removal of myxomatous debris. Using these techniques, 17 patients have been treated successfully at the Brigham and Women's Hospital. There were no operative deaths and no significant perioperative morbidity. Fifteen patients are asymptomatic, and one patient has minimal shortness of breath an average of 57 months after surgery. One patient died of leukemia 15 months postoperatively. No recurrent myxomas have been identified, and no patients have symptoms of other cardiac disease. PMID- 2980000 TI - Reconstructive techniques for rheumatic aortic valve disease. AB - Aortic valve reconstruction is still at an early stage of development. We report techniques that can be applied in the repair of rheumatic aortic valve disease in patients with concomitant mitral valve disease. The techniques described are: (1) commissurotomy, (2) cusp free edge unfolding, (3) annuloplasty, and (4) supraaortic crest enlargement. Fifty patients operated on between January 1974 and January 1986 with a hospital mortality of 6% were followed for a mean period of 7.7 years. Reoperation due to failure of mitral surgery was required in twelve patients, tricuspid regurgitation in two, and significant aortic regurgitation in four. These techniques, although applicable to a limited number of patients, have enabled us to avoid the problems of aortic valve replacement in many patients. PMID- 2980001 TI - The Ross operation: the autologous pulmonary valve in the aortic position. AB - Aortic valve replacement (AVR) with a pulmonary valve autograft (PVA) was first reported by Donald N. Ross (DNR) in 1967. The expectation of this procedure was to avoid degenerative changes seen in other biological tissue valves such as calcification, attenuation, and rupture of the leaflets. Recent reports by the original investigator's group have confirmed the lack of degenerative changes in PVA. To corroborate their conclusions, the fate of 12 patients undergoing AVR with PVA by one of us (LGL) has been ascertained. From March 1969 to June 1971, 12 patients underwent AVR with PVA. The right ventricular outflow tract (RVOT) was reconstructed with an aortic homograft valved conduit. The mean age was 42.7 years (range 21 to 52 years). The mean follow-up for 11 hospital survivors is 12.4 years. Three PVA have been replaced; one following infective endocarditis at 13 years, and two at 15 and 73 months due to technical malalignment. There was no evidence of PVA degeneration during histological examination of these explanted PVAs. Six patients are alive and retain the original PVA at 12 years (55%). This analysis corroborates the conclusions of the DNR report and strongly suggests an immunological mechanism in the process of calcification of other biological tissue valves. The Ross operation is advocated for AVR in young patients as valve durability is of paramount importance especially in this group. PMID- 2980002 TI - Fontan hemodynamics. AB - Right atrial-pulmonary artery connection places the pulmonary circulation in series with the systemic circulation rather than the single ventricular "parallel" circulatory arrangement that usually is present prior to repair. The accompanying central cardiac shunt and volume overload physiology are eliminated. Favorable changes in ventricular dimension, ventricular wall stress, cardiovascular efficiency, relative systemic perfusion, and arterial oxygen saturation should result. The ongoing myocardial injury associated with the single-ventricle volume overload is presumably arrested and repair is initiated to a variable degree. Some candidates for right atrial-pulmonary artery connection may not benefit from repair because of irreversible ventricular injury. More accurate indices of systolic and diastolic ventricular function should be applied to this difficult group of borderline patients to further define potential for myocardial recovery and, therefore, candidacy for Fontan repair. PMID- 2980003 TI - Percutaneous transluminal coronary angioplasty. AB - The relief of coronary obstruction by surgical grafting was the first effective treatment to be directed at the cause of ischemic heart disease. PTCA represents the second major step in relieving coronary stenosis. It seems timely to review where this second step has led in order to understand how percutaneous transluminal coronary angioplasty (PTCA) relates to surgery and to understand future implications of this procedure. This review will reflect many of the authors biases and prejudices derived from their experience at Mayo Clinic. It will also be a somewhat practical assessment. Such a pragmatic approach can be defended because PTCA itself is built on pragmatism. It is more of the world of craftsmen than of the world of scientists. We are still waiting for science to "catch up" to help solve important remaining problems such as the issue of acute rethrombosis and restenosis. Our ability to review the Mayo Clinic experience is made possible only by having a dedicated team of colleagues in the catheterization laboratory and a dedicated support group managing our PTCA Registry. Experience with other large registries (the NHLBI CASS Registry and the PTCA Registry) has taught us that this is a valuable method of collecting and reviewing the experience with a new procedure as it develops. This approach is likely to be more widely applied in the future. PMID- 2980004 TI - A simple technique for covering bilateral IMA. PMID- 2980005 TI - A simple technique for aortic valve replacement using freehand allografts. AB - Given the recent resurgence of interest in the use of "fresh" and cryopreserved allograft valves for aortic valve replacement, the fact that many cardiac surgeons were not exposed to the operative techniques involved in freehand implantation of allograft valves during their residency training, and the paucity of teaching materials that clearly portray such techniques, details of a simplified method of subcoronary, freehand allograft valve implantation in the aortic position are described and illustrated. PMID- 2980007 TI - Mortality data for coronary bypass surgery and the utilization of this data to compare institutions. PMID- 2980006 TI - Mycotic disruption of aortic cannulation site. AB - Two cases of successful repair of leaking mycotic false aneurysms of the ascending aorta from the aortic cannulation site, secondary to mediastinal infection following open heart surgery are described. Institution of cardiopulmonary bypass via the femoral vessels, rapid sternotomy, and fingertip control of the aortic hemorrhage permitted primary repair of the disruption. PMID- 2980008 TI - Perioperative dissection of the ascending aorta: types of repair. AB - Iatrogenic aortic injury occurring during either coronary bypass grafting or valve replacement is a well-recognized complication of cardiac surgery. We retrospectively reviewed our experience and found 11 cases occurring in a case load of 8,945 hearts (incidence of 0.12%). All 11 cases were repaired, with 10 patients surviving. The type of repair used usually was determined by when the diagnosis was made. When an intraoperative diagnosis was made, a local repair was done in four of six cases. If a postoperative diagnosis was made, then all five patients needed the ascending aorta replaced. With early diagnosis and rapid repair, good surgical results can be achieved. PMID- 2980009 TI - Surgical considerations for avoiding disc interference based on a ten-year experience with the Medtronic Hall heart valve. AB - All low profile valves are susceptible to problems related to implant technique, especially immobilization of the occluder. Because the disc-valve-ring clearance may be only a fraction of a millimeter, this complication is a small yet ever present possibility of which the surgeon must be aware. Special care must be taken when implanting low profile valves to eliminate the possibility of disc interference caused by suture ends or intracardiac structures. The surgeon can minimize the risk of disc immobilization by careful attention to precautionary surgical measures described herein. PMID- 2980011 TI - Cardiac neoplasms: current diagnosis, pathology, and therapy. AB - Cardiac neoplasms remain enigmatic because of their rarity and protean clinical manifestations. Although surgery for benign cardiac neoplasms seems straightforward, certain difficulties in diagnosis and extirpation often exist. Newer diagnostic modalities including intraoperative transesophageal echocardiography seem promising and may provide better planning for the surgeon. Moreover, cardiac transplantation has provided relief for some patients with irresectable benign neoplasms. As ventricular arrhythmias may emanate from benign tumors, electrophysiologically guided extirpation may provide helpful control. Malignant cardiac tumors remain the true challenge, whether primary or metastatic, as little surgical or medical progress has resulted. This review examines nearly all diagnostic, pathological, histologic, and therapeutic aspects of benign and malignant cardiac tumors. Moreover, the various surgical approaches necessary for removal of resectable neoplasms are detailed. PMID- 2980010 TI - Treatment of the infected cardiac suture line. AB - After combined left ventricular aneurysmectomy and myocardial revascularization, four patients developed infection at the cardiac suture line. The infected cardiac suture line causes a variety of complications including cardiocutaneous fistula, erosion of pulmonary tissue, or pseudoaneurysm. The diagnosis is made by sinogram and left ventricular angiography, which is mandatory in all patients with suspected infection to guide the surgical approach. Once the diagnosis is made, aggressive and prompt surgical treatment is necessary to remove all infected material. A left anterolateral thoracotomy in the fifth or sixth intercostal space is the preferred approach. In the presence of a pseudoaneurysm, femoro-femoral bypass is required to reopen the left ventricle without cross clamping the aorta. The septic material is removed, left ventricular wall and pericardial fibrous scar are closed, and extensive antibiotic treatment administered. All four patients survived and are free of complications two to three and a half years postoperatively. PMID- 2980012 TI - Pericardial patch repair of aortic cusp perforation caused by cardiac catheterization. AB - A case of acquired aortic insufficiency due to perforation of the aortic leaflet at the time of the diagnostic heart catheterization is presented. The situation was remedied by repairing the perforated leaflet using a pericardial patch. PMID- 2980013 TI - A technique to facilitate mitral valve replacement. AB - Replacement of the damaged mitral valve with a biological or mechanical prosthesis occasionally can be difficult. A technique is described that facilitates (a) exposure of the mitral valve, (b) safe excision of the mitral and subvalvular apparatus, (c) good access to the annulus for placement of interrupted sutures in an operative setting that provides good venous return, adequate myocardial protection, and the opportunity for complete removal of air. The procedures described are particularly helpful in reoperative mitral valve surgery when adhesions may limit exposure. The principles have been derived from the need for superb mitral valve exposure during reparative surgery. Perivalvular leak and myocardial rupture have been virtually eliminated by adoption of the principles outlined. PMID- 2980014 TI - Modified Fontan procedure: atrial baffle and systemic venous to pulmonary artery anastomotic techniques. AB - Two technical modifications to the modified Fontan procedure are presented. Systemic venous to pulmonary artery continuity is achieved by superior vena cava (SVC) division and end to side anastomosis to the right pulmonary artery (RPA), particularly following a right Blalock shunt, or by RPA division and anastomosis to the SVC, particularly in the presence of RPA stenosis. Intra-atrial partitioning is achieved by a systemic venous baffle rather than a pulmonary venous baffle. This is particularly useful in the presence of left atrioventricular valve atresia, but may be a preferable technique with double inlet single ventricle or single ventricle with common AV valve. These techniques were applied successfully to 10 of 12 children with various forms of single ventricle, including 5 with left AV valve atresia or stenosis. PMID- 2980015 TI - Correction of tetralogy of Fallot with combined transatrial and pulmonary approach: experience with 184 consecutive cases. AB - The authors present their technique and results for complete correction of Fallot's tetralogy by closure of the ventricular septal defect by the transatrial approach, and resection of the infundibular hypertrophy by a combined approach through the tricuspid and pulmonary valve orifices. Right ventricular pressures were often high at the end of operation due to a residual muscular gradient but the outcome of these postoperative gradients was similar to that of valvular pulmonary stenosis with an intact ventricular septum. PMID- 2980016 TI - Surgical treatment of ascending aortic pathology. AB - Among the first 10,200 valvular replacements performed in our unit, 288 complex repairs of the ascending aorta were done for various aortic pathology. Aneurysms of the ascending aorta were the most frequent; 53 supracoronary artery aneurysms with aortic valvular insufficiency were treated by the separate replacement of the aortic valve and the supracoronary ascending aorta; 206 annulo-aortic ectasia had total and combined replacement of the ascending aorta and the aortic valve with a personal modification of the Bentall's technique using an 8-mm diameter Dacron graft to perform the reimplantation of the coronary arteries on the composite aortic grafts. The operative mortality for the first 100 patients was 4% and for the entire 206 patients, 6%. Late mortality during a follow-up period ranging from 18 months to 8 years was 11%. The actuarial survival rate at 8 years is 75%; 25 patients restudied by angiography demonstrated satisfactory results with neither stenosis nor aneurysm on the coronary graft but a recurrent or persisting chronic distal aortic dissection in four patients. In 26 cases of aortic valvular endocarditis, large abscesses of the aortic annulus involved the aortic root. In 11, the aortic repair consisted of the insertion of a subcoronary valved conduit (two early deaths, two late deaths, one reoperation, seven good results--maximum follow-up of eight years). Twelve patients had a supracoronary valved conduit with four early deaths, one late death, and two reoperations; seven are alive and well, two to six years later. Three patients previously operated had a left ventricular abdominal aorta valved conduit; two of them are alive and well up to six years later. In three patients with iterative aortic paravalvular leak (recurring three or four times), ablation of the aortic insufficiency was obtained by interposition of a composite valved graft in the ascending aorta. PMID- 2980017 TI - Mechanical support for postcardiotomy heart failure. AB - Cardiac failure remains a life-threatening complication for certain patients undergoing intracardiac repair. Despite improvements in surgical techniques, methods of myocardial protection, and postoperative care, patients are frequently at risk to develop postoperative low output syndrome. Approximately 1% of cardiac surgical patients cannot be weaned from extracorporeal circulation in spite of adequate volume loading, the use of inotropic support, and initiation of intraaortic balloon pumping. In these cases, ventricular assist devices (VAD) can mechanically aid the failing heart and reverse the low output state. The concept of mechanical support for the failing left ventricle was first proposed by Clauss et al. in 1961. By 1968, Kantrowitz and associates had developed and refined the first intraaortic balloon pump (IABP). Through the efforts of Moulopolous and others, this device evolved into the present-day intraaortic balloon pump (IABP). Clinical evidence for the efficacy of left ventricular assist devices (LVAD) remained questionable until 1980, when the National Heart, Blood and Lung Institute evaluated short-term LVADs by comparing various types of mechanical aids. This report focused attention primarily on the failing left ventricle (LV). As the use of inotropic support, intraaortic balloon pumping, and LVADs improved, a small group of patients emerged who could not be separated from extracorporeal circulation due to a failing right ventricle. The failing right ventricle emerged as a unique clinical entity similar to postcardiotomy left ventricular failure that also benefited from mechanical cardiac assistance. Current therapy at major centers incorporating mechanical assist devices is based on the premise that the low output state will allow the failing heart to recover from a reversible injury. The frequent occurrence of postcardiotomy ischemia may be due to several factors such as poor myocardial protection, overdistension of the LV, emboli, coronary spasm or technical problems. Whatever the etiology, the end product of cardiac failure is a demand for oxygen consumption that cannot be met, thus leading to cardiac demise. PMID- 2980018 TI - Clinical experience with expanded polytetrafluoroethylene Gore-Tex surgical membrane for pericardial closure: a study of 110 cases. AB - Complete closure of the pericardium after cardiac operations has the advantage of avoiding injury of the heart and great vessels during reoperation. Between 1985 and 1987, the pericardium was closed with Gore-Tex Surgical Membrane (SM) in a selected series of 110 patients 1 month to 76 years of age. Fifty-three patients had congenital heart lesions and 57 patients had acquired heart disease. Overall hospital mortality was 3/110 cases. In no instance was there a relationship between occurrence of death and pericardial closure with SM. There was one episode of cardiac tamponade on the seventh postoperative day. One patient developed fever and leukocytosis due to a mediastinal hematoma. During a mean follow-up of 15 months, four patients had to be reoperated upon three, four, eight weeks, and eight months after primary operation. The anterior wall of the heart had no adhesion with the SM and the other parts of pericardium could be dissected easily. Scanning electron microscopic examination of the explanted SM patches showed neither cellular ingrowth nor immunocompetent cellular elements. The Gore-Tex Surgical Membrane has the advantages of easy availability and lack of reaction between its surface and the epicardium and pericardium. We believe its routine use should be encouraged in patients with high probability of reoperation after repair of complex cardiac anomalies, implantation of bioprostheses, coronary revascularization for one- or two-vessel disease, and repair of degenerative disease of the ascending aorta. PMID- 2980019 TI - The superior approach for mitral valve replacement. AB - A consecutive series of 98 patients ranging from 2 1/2 to 79 years of age underwent mitral valve replacement using the superior approach that entails an atriotomy done between the superior vena cava and the ascending aorta prolonging it into the left superior pulmonary vein. The technique opens the roof of the left atrium without dissection, frequently without the need for double cannulation of the right atrium. No mobilization of the heart is involved, which is left in its normal position. There were 62 patients undergoing replacement alone, 22 combined with coronary bypass surgery, 10 with simultaneous aortic valve replacement, 2 with coronary bypass and left ventricular aneurysm resection, and 2 others combined with ventricular septal defect (VSD) closure and placement of an extracardiac conduit. Forty-six were done for stenosis and 52 for regurgitation. No technical difficulties were encountered, and the valve can easily be replaced through an incision slightly larger than the diameter of the prosthesis being implanted. Since the heart is not mobilized, the ventricles remain immersed in the cold topical solution (4 degrees C) in addition to the administration of cardioplegia for myocardial protection. The access is simple and offers better exposure for the assisting surgeon than the usual inter-atrial groove approach. Important steps of the technique are clarified. PMID- 2980020 TI - Valvular-ventricular interaction: the importance of the mitral chordae tendineae in terms of global left ventricular systolic function. AB - While conventional mitral valve replacement (MVR) for patients with chronic mitral regurgitation has been associated with relatively high operative mortality rates and incidence of late postoperative left ventricular (LV) failure and death, chordal-sparing mitral valve operations (valve repair/reconstruction or MVR with preservation of the chordae tendineae) subjectively appear to portend lower operative morbidity and mortality rates, better functional results, and improved long-term survival rates. Such empirical clinical observations have provided the basis for the concept of valvular-ventricular interaction, namely, that the intact mitral chordae are important mediators of more efficient and forceful ventricular contraction that enhances LV performance. This paper reviews the pertinent basic physiology and dynamics of the chordae tendineae and papillary muscles and examines critically the available experimental and clinical data regarding valvular-ventricular interaction. The problems inherent in quantifying LV contractility are central to this discussion and are also examined. While earlier experimental studies have produced conflicting results, more recent experiments utilizing load-independent measures of ventricular performance (particularly in isovolumic preparations) have conclusively demonstrated the importance of chordal integrity for optimal LV systolic function in normal animal hearts. The balance of the clinical evidence is also suggestive (although by no means conclusive) regarding the importance of valvular ventricular interaction. Recent experimental evidence suggests that the mitral chordae enhance LV systolic function by means of regional afterload reduction. The mechanism(s) responsible for valvular-ventricular interaction, however, remains incompletely characterized at the present time, which underscores the urgent need for further experimental and, most importantly, clinical studies. PMID- 2980021 TI - Sternotomy closure with Parham bands. AB - The closure of a median sternotomy incision requires secure bony approximation to prevent postoperative pain, sternal click, and/or nonunion of bone. The standard technique of sternotomy closure involves the use of stainless steel wires for reapproximation of the sternum. These wires occasionally break or pull through bone, resulting in instability of either a portion of the sternum or the entire sternum. Presented here is our technique for sternotomy closure that provides secure closure with reduced postoperative morbidity. PMID- 2980022 TI - Preoperative saphenous vein mapping for coronary artery bypass. AB - Saphenous vein mapping by real-time duplex ultrasound scanning allows for the confirmation of the presence of patent vein and determination of the anatomical course of the vein prior to aortocoronary bypass procedures. This technique has proven to be of value in patients whose veins are not evident by physical examination and in those whose length of conduit may be limited by previous vein stripping or thrombophlebitis. PMID- 2980023 TI - Use of the valvular resistance in the separation of normal and stenotic Hancock mitral valves. AB - Recent data suggests that the pressure-flow relationship for normal bioprosthetic mitral valves is linear. If this is correct, the valve resistance may provide a better indicator of normal mitral function than the Gorlin valve area. We compared the Gorlin valve area to the valve resistance (calculated as flow/pressure) in order to determine which better separated normal and stenotic Hancock mitral valves. Measurements were made using left atrial and left ventricular catheters in 42 patients undergoing Hancock mitral valve replacement. Patients were studied during pacing and isoproterenol infusion for a total of 141 measurements. Stenotic Hancock mitral valve hemodynamics were obtained at cardiac catheterization from eight patients who were studied at rest and during atrial pacing and from an additional eight patients culled from the literature (a total of 23 stenotic measurements). The Gorlin valve area ranged from 1.1 to 4.4 cm2 for the normally functioning Hancock valves and from 0.4 to 1.54 cm2 for the stenotic valves. Six measurements in patients with confirmed stenotic valves yielded Gorlin areas larger than the lowest area found in the normal valves and no value of the Gorlin valve area correctly classified all of the normal and the stenotic valves.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980025 TI - Leukocytes, platelets, and surface microstructure of spontaneously degenerated porcine bioprosthetic valves. AB - The microstructure of 33 spontaneously degenerated porcine bioprosthetic valves was assessed by scanning electron microscopy in order to gather insight regarding the degenerative process. Twenty-four mitral and 9 aortic valves were removed from 32 patients. The duration of insertion was 7.7 +/- 2.4 years (mean +/- SD). All valves showed denudation of endothelial cells and exposure of the subendothelial fibrous tissue. Fibroblastlike cells were occasionally seen. Platelet deposits were observed on 22 of 33 valves (67%). Leukocytes were observed on the surface of 27 of 33 valves (82%). Mononuclear leukocytes were the most common category of cells (66%). Crystalline material was present on the surface of some leukocytes, suggesting that they may serve as a nidus for calcification. Transmission electron microscopy showed leukocytes in the process of phagocytizing collagen fibers. Macrophages, by exerting their scavenger function seem to contribute to destruction of the collagen framework of the valves. Whether the observed lymphocytes and plasma cells reflect an immunological involvement is unclear. PMID- 2980024 TI - The cryopreserved homograft valve in the pulmonary position: early results and technical considerations. AB - Since September, 1985, 20 patients have undergone implantation of a homograft valve in the pulmonary position (16 pulmonary, 4 aortic). There were 11 primary operations and 9 reoperations. In 7 of 11 primary operations the homograft valve was utilized as a composite conduit with a short Dacron extension. In four of five reoperations for a failed porcine valved conduit, a composite homograft conduit was used. Four patients underwent implantation of a free homograft in a previously repaired right ventricular outflow tract (RVOT). Age ranged from 15 days to 22 years. There was one operative death (5%), a seven-week-old infant with truncus arteriosus. Long-term follow-up ranges from 1 to 30 months. Clinical performance has been satisfactory in 18 of 19 patients. One patient undergoing free implantation of a pulmonary valve in the RVOT required replacement at 18 months with a porcine valve. In this patient, pulmonary insufficiency was caused by distortion of the annulus secondary to dilatation and pulmonary hypertension. Nine of 18 survivors do not require medication. Eleven of 18 have trivial to mild pulmonary insufficiency murmurs without symptomatology. The homograft valve is extremely useful in reconstruction of the right heart, however, early insufficiency murmurs have been noted. Distortion of the valve annulus may contribute to the early onset of a benign insufficiency murmur. Residual distal obstruction or pulmonary hypertension may be a contraindication to the use of a free homograft in the orthotopic position. PMID- 2980026 TI - Prognostic importance of viability and a study of a second set allograft valve: an experimental study. AB - An experimental study of transplantation of first and second set aortic valve allografts in heterotopic infrarenal aortic position in inbred rats with varying histocompatibility is described. On the 20th and 50th postoperative days, there was deposition of donor-specific antibodies on the allograft endothelia of the weakly allogeneic, non-MHC group, as evidenced by immunofluorescence studies, whereas the nonviable grafts showed no identifiable antibodies on their endothelia. The endothelia were mostly lost at day 100; previous focal mononuclear cellular infiltrates disappeared. The ground substances, however, were maintained until the 250th day. In the nonviable allografts, the media was completely acellular on the 50th day with collagen disintegration and changes in ground substance on the 100th day. Loss of endothelia and replacement by fibrin deposits with transient focal cellular infiltrations were the most significant early microscopic findings. Acellularity, fibrous neointima, and leaflet thickening due to varying degrees of fibrosis with changes in ground substance and focal infiltrations of macrophages around degenerated collagenous matrix of aortic valve allografts are the significant late changes. The second set allograft valves showed no difference in the rate or type of healing, but early degeneration while the second set skin grafts underwent accelerated rejection, thus confirming prior sensitization. This finding confirms the weak antigenicity of cardiac valve allografts. Therefore, the use of cardiac valve allografts for secondary valve replacement might be favorable if properly used. Limitation of antigen incompatibility by considering at least the ABO matching and reduction of a recipient's immunological reactivity might be a proper step to achieve a longer survival of viable allografts. PMID- 2980027 TI - A technique for selective graft perfusion during aortocoronary bypass. PMID- 2980028 TI - A study of the cells in the explanted viable cryopreserved allograft valve. AB - From June 1975 to December 1987, 231 patients underwent aortic valve replacement with a viable cryopreserved allograft aortic valve. Throughout this era, a uniform procurement and preservation was used to maintain leaflet fibroblast viability. The allograft valve was obtained from coroner's autopsies within 24 hours of death, and more recently from organ donors, incubated for 24 hours in low dose antibiotic solution followed immediately by cryopreservation (mean time interval 39 hours after donor death). Viability was ensured by monitoring glucose utilization of the aortic and pulmonary valves and by demonstrating fibroblast growth in tissue cultured from the pulmonary valve. A uniform protocol for valve preparation was used during the entire experience. Nine allograft aortic valves have been obtained by eight reoperations (two were for leaflet degeneration) and one autopsy. The time intervals from implantation to explantation were 2 months, 10 months, 20 months, 22 months, 2.2 years, 5 years, 8.3 years, 9.2 years, and 10.8 years. Histologic examination of the leaflet tissue disclosed a variable degree of cellularity, ranging from a highly cellular matrix (9.2 years) to minimal cellularity (20 months). Within the same valve (10 months), one leaflet was completely acellular with a moderate degree of cellularity in the other two leaflets. The competent valve recovered at autopsy (8.2 years) was essentially acellular. Fibroblasts could consistently be cultured from leaflets in which viable cells were seen histologically. Chromosomal analysis of cultured cells from a valve leaflet (9.2 years) that was implanted with a donor and recipient sex mismatch demonstrated persistence of donor cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980029 TI - Long-term results of the viable cryopreserved allograft aortic valve: continuing evidence for superior valve durability. AB - From December 1969 to May 1975, 124 patients underwent aortic valve replacement with an allograft aortic valve sterilized by incubation in a low dose antibiotic solution and stored by refrigeration at 4 degrees C (4 degrees C stored valve group). From June 1975 to December 1987, 231 patients received an allograft aortic valve, sterilized by the same low dose antibiotic solution, but stored by cryopreservation in liquid nitrogen at -196 degrees C (cryopreserved valve group). The 4 degrees C stored valves were essentially nonviable, whereas the cryopreserved valves were viable at implantation. Of the 355 aortic valve replacements, associated procedures were performed in 127 patients. The 30-day mortality was 8.9% (confidence limits [C.L.] 6.2% ... 12.3%) (4 degrees C stored) and 4.8% (C.L. 3.3% ... 6.7%) (cryopreserved). Actuarial survival was similar in both groups, being 71% and 67% at 10 years in the 4 degrees C stored and cryopreserved valve groups, respectively (P = .18). The probability of a thromboembolic event was low, but appeared higher in the 4 degrees C stored valve group (actuarial freedom at 10 years, 90%) than the cryopreserved valve group (actuarial freedom at 10 years, 98%) (P = .01) probably related to associated mitral valve surgery. The actuarial freedom from allograft valve endocarditis at 10 years was 94% and 95% for the 4 degrees C stored and cryopreserved valve groups, respectively (P = .23). Reoperation was undertaken in 34 patients in the 4 degrees C stored group and 12 patients in the cryopreserved valve group for leaflet degeneration, endocarditis, or technical reasons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980030 TI - Homograft replacement of single aortic valve cusps: 22 years follow-up. AB - Single aortic valve cusp replacement with fresh aortic homografts were performed in four patients from November 1966 to April 1970. All had preoperative aortic insufficiency due to destruction of a single cusp, with the remaining two cusps structurally and functionally intact. Replacement was performed using the noncoronary cusp from fresh aortic valve homografts. Patient one had homograft cusp replacement of the left coronary cusp at age 13 due to bacterial endocarditis. She recently underwent mitral valve replacement; study and inspection of the aortic valve showed it to be structurally and functionally intact 22 years following homograft cusp replacement. Patient two sustained a gunshot wound perforating his right coronary cusp. He underwent single cusp replacement in January 1967. Currently, he is asymptomatic and has been without evidence of significant aortic valve dysfunction for the past 21 years. Patient three had destruction of the noncoronary cusp due to endocarditis and had homograft cusp replacement in October 1967. Four and a half years later he developed recurrent endocarditis, presented with severe congestive heart failure due to aortic insufficiency, and died. Patient four had bacterial endocarditis affecting the right coronary cusp and had replacement in April 1970. In February 1980, he underwent aortic valve replacement for aortic stenosis. Inspection revealed calcification of the other two cusps with the homograft cusp structurally normal. In summary, follow-up of homograft replacement of single aortic valve cusps from 4 1/2 to 22 years, shows it to be a durable and feasible alternative to prosthetic valve replacement. PMID- 2980031 TI - Morbidity following the Ross operation. AB - Aortic valve replacement (AVR) with a pulmonary valve autograft (PVA) was first reported by Donald N. Ross in 1967. The expectation of this procedure was to avoid degenerative changes seen in other biological tissue valves such as calcification, attenuation, and rupture of the leaflets. Recent reports by the original investigator's group have confirmed the lack of degenerative changes in PVA. To corroborate their conclusions, the fate of 12 patients undergoing AVR with PVA by Dr. Gonzalez-Lavin has been ascertained. From March 1969 to June 1971, 12 patients underwent AVR with PVA. The right ventricular outflow tract (RVOT) was reconstructed with an aortic homograft valved conduit. The mean age was 42.7 years (range 21 to 52 years). The mean follow-up for 11 hospital survivors is 12.4 years. Three PVAs have been replaced; one following infective endocarditis at 13 years, and two at 15 and 73 months due to technical malalignment. There was no evidence of PVA degeneration during histologic examination of these explanted PVA. Six patients are alive and retain the original PVA at 12 years (55%). This analysis corroborates the conclusions of Dr. Ross and strongly suggests an immunological mechanism in the process of calcification of other biological tissue valves. The Ross operation is believed to be the preferred method of AVR in young patients. PMID- 2980032 TI - Homograft valve preparation and predicting viability at implantation. AB - An experimental study was performed using C14 proline uptake in order to: (1) assess the effects of current sterilization and storage methods on fibroblast viability, and (2) establish a control tissue that could be used to determine viability of each homograft valve at the time of implantation in the clinical setting. The results were expressed as disintegrations per minute per milligram of tissue (DPM/mg). Swine aortic (AV), pulmonary (PV), and tricuspid leaflets (TV), and adjacent AV and PV arterial wall were procured sterile and subjected to routine sterilization and storage. Thirty samples of AV were analyzed for incorporation of labeled proline at procurement (208 +/- 7 DPM/mg), following 48 hour antibiotic exposure (87 +/- 6 DPM/mg, P less than .0001), and following controlled rate cryopreservation and storage for 12 days at -80 degrees C (78 +/- 8 DPM/mg, P = .42). Proline uptake of the other tissues at the same intervals disclosed that only the TV resulted in the same degree of viability at implantation (AV 78 +/- 8, PV 68 +/- 3, TV 75 +/- 2, P = NS). The homograft valves were obtained under sterile conditions from brain dead, multiorgan donors (homovital). It has been postulated that these valves are sterile and ready for implantation. Of 17 homovital valves cultured at procurement, 9 had positive cultures within 48 hours (53%). We conclude that: (1) the TV can be processed as a control tissue with each homograft and then utilized to predict viability at the time of implantation in the clinical setting; (2) antibiotic exposure is an essential step in the preparation of all homografts, however, modification of the antibiotic solution is necessary. PMID- 2980033 TI - Pulmonary valve autotransplantation (the Ross operation). AB - In 1967, homografts had been in place five years and the early nonviable and natant freeze-dried valves were showing irrefutable signs of degeneration and calcification. For this reason, in 1967, we moved to reserving both viability and tissue integrity by cryopreservation. To overcome the problems of procurement, the next logical step was to transfer the patient's own pulmonic valve to the aortic area. In the past twenty years, 249 operations have been performed. Of the 249 patients, there have been 16 operative deaths (6.5%), but no operative deaths in the past ten years. There have been no details related to primary tissue failure in the aortic area. There have been 36 reoperations (14.4%). PMID- 2980034 TI - The Carpentier-Edwards standard porcine bioprosthesis: long-term evaluation of the high pressure glutaraldehyde fixed prosthesis. AB - The Carpentier-Edwards standard porcine bioprosthesis was implanted in 1,000 operations (988 patients with 1,092 valves) between 1975 and 1981. The mean age of the patients was 56.8 years (range 8 to 85 years). During the years 1975 and 1981, approximately 97% of the total valvular surgery population received the prosthesis. The early mortality was 7.8% (including patients with concomitant procedures including coronary artery bypass and ascending aortic aneurysm resection). Late mortality was 3.8% per patient-year. The total cumulative follow up was 5,937 years. Thromboembolism (TE) was 1.3% per patient-year (fatal 0.4%/patient-year) (minor 0.4%; major 0.9%); antithromboembolic therapy-related hemorrhage (ATH) 0.5% (fatal 0.1%); prosthetic valve endocarditis (PVE) 0.5% (fatal 0.2%); periprosthetic leak (PPL) 0.4% (fatal 0.2%); clinical valve dysfunction (CVD) 0.2% (fatal 0.02%); and structural valve deterioration (SVD)/primary tissue failure 1.6% per patient-year (fatal 0.2%/pt-yr). Thromboembolism and SVD occurring primarily between the sixth and tenth year of evaluation, were the significant complications. The overall patient survival was 60.5% +/- 2.4% at 10 years. The patients were classified as 93.5% NYHA functional Class III and IV, preoperatively and 93.1% Class I and II, postoperatively. The freedom at 10 years from TE was 82.9% +/- 2.7%; SVD 76.9% +/- 2.9%; and reoperation 70.8% +/- 3.1%. Freedom from all valve-related complications at 10 years was 54.3% +/- 3.1%; valve-related mortality 86.4% +/- 2.3%; mortality and reoperation (valve failure) 64.0% +/- 3.2%; mortality and residual morbidity (treatment failure) 82.3% +/- 2.6%; and mortality, residual morbidity, and reoperation (valve failure and dysfunction) 60.6% +/- 3.2%. There were 61 valve related deaths of a total 352 deaths (early 7, late 54) (TE 21; ATH 7; CVD 1; PVE 12; PPL 9; and SVD 11). Valve-related reoperations were performed in 128 patients (TE 3; CVD 4; PVE 14; PPL 17; and SVD 90). The standard Carpentier-Edwards porcine bioprosthesis has provided very satisfactory clinical performance and afforded patients excellent quality of life. Primary tissue failure is the significant long-term complication. PMID- 2980035 TI - The standard Hancock porcine bioprosthesis: overall experience at the University of Padova. AB - All patients undergoing aortic (AVR, n = 196), mitral (MVR, n = 502), and mitral aortic (MAVR, n = 71) valve replacement with a standard Hancock porcine bioprosthesis (HPB) from 1970 to 1983 were reviewed. A total of 665 patients discharged were followed for 5,099 patient-years with an actuarial survival at 15 years of 52% +/- 4.5%, for MVR, 37% +/- 14% for AVR, and at 12 years of 52 +/- 7.4% for MAVR. Embolic episodes occurred in 9 patients after AVR (0.7% +/- 0.2% pt-yr), in 61 after MVR (1.7% +/- 0.2% pt-yr), and in 6 after MAVR (1.7% +/- 0.7% pt-yr); actuarial freedom from emboli at 15 years is 91% +/- 3.5% after AVR, 79% +/- 14% after MVR, and at 12 years is 87% +/- 5% after MAVR. Reoperation because of primary tissue failure (PTF) was performed in 47 patients with AVR (3.9% +/- 0.5% pt-yr), 91 with MVR (2.6% +/- 0.3% pt-yr), and in 13 with MAVR (4.1% +/- 1.1% pt-yr); actuarial freedom from PTF at 15 years is 41% +/- 5.5% after MVR, 37% +/- 10% after AVR, and at 12 years is 49% +/- 13% after MAVR. After AVR and MVR, freedom from PTF is significantly better for patients over 50 years of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980036 TI - The INTACT bioprosthesis--early results. AB - The INTACT (previously Xenotech) valve incorporates three major departures from previously available bioprostheses. The following innovations are aimed at reducing calcification and improving durability: (1) the stent has a high profile configuration and the sinuses are deeply excavated. This should maintain normal coaption of the leaflets and prevent cusp abrasion; (2) stress-free fixation preserves the structural characteristics of the leaflets; (3) the valve is treated with toluidine blue, which occupies the calcium binding sites. During a 36-month period 185 valves were implanted in 167 patients. The procedure used most often was isolated mitral valve replacement. Twenty-six patients were under the age of 20. There were 8 hospital deaths (5% mortality) and 12 late deaths. Two of the hospital deaths were the result of ventricular perforation, and it is believed that the high profile of the stent contributed. In the group aged less than 20 years, there have been three episodes of valve calcification giving a 3 year actuarial rate for calcification of 22%. In the adult group, there have been no valve failures, and postoperative catheterization studies in this group demonstrated satisfactory hemodynamics. PMID- 2980037 TI - Mid-term results of the Liotta-Bioimplant low profile bioprostheses. AB - Low profile bioprostheses are particularly useful for certain anatomical conditions. In some patients with rheumatic aortic insufficiency, an important dilatation of the aortic annulus is present, even when the subcoronary aorta is not enlarged. In these cases, the bioimplant heart valve with its low height avoids any threat to the aortic wall or to the coronary ostia. Frequently, the left ventricular cavity is not enlarged in patients with mitral stenosis. The characteristic low profile of this valve avoids left outflow obstruction as well as traumatism of the left ventricular wall. In the tricuspid position, this design is particularly useful because it leaves the right ventricular cavity totally free. From February 1981 to December 1983, 198 bioimplant (LIOTTA) low profile bioprostheses were implanted in 184 patients. There were 63 aortic (AVR), 101 mitral (MVR), 14 mitral and aortic (MAVR), and 6 tricuspid (TVR) valve replacements. Early mortality (30 days) was 6% (AVR = 1.6%; MVR = 8.9%; MAVR = 7.1%; TVR = 0). The 181 survivors were followed over a period of 3-84 months (643 patient-years). The thromboembolic complication rate was low (0.7%/patient-year) and 96.9% +/- 1.4% of patients were free of thromboembolism (AVR = 98.2% +/- 1.7%; MVR = 97.6% +/- 1.7%). Five years after implant, 91.7% +/- 3.2% of patients were free of valve failure (AVR = 93.7% +/- 4.4%; MVR = 88.8% +/- 5.2%). Actuarial analysis shows an expected survival at 5 years (average) of 87.2% +/- 3.4% (AVR = 87.4% +/- 6%; MVR = 87.3% +/- 4%) with an actuarial rate of freedom from reoperation of 87.5% +/- 3.5%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980038 TI - Clinical comparison of mitral valve replacement using porcine, Starr, and Bjork valves. AB - The choice of a cardiac prosthesis for mitral valve replacement remains controversial, and thromboembolic complications are still a major cause of morbidity and mortality in patients with mechanical valves. Because of this, permanent anticoagulation with its risks and constraints on daily life is necessary. Bioprostheses, however, are associated with a lower rate of thromboembolic events. Therefore, the need for long-term anticoagulation is minimized. These advantages are counterbalanced by the limited durability of tissue valves. In an effort to give some perspective to this balance, we compared the long-term results of three commonly used mitral valve prostheses. Three hundred patients operated on in the same institution January 1974 to December 1978 form the basis of this evaluation. PMID- 2980039 TI - The Carpentier-Edwards bioprosthesis: a comparative study analyzing failure rates by age. AB - Bioprosthetic valve durability and the significance of patient age at implantation have received much attention recently. Indications and/or contraindications for implantation of the bioprosthesis in the very young and in the elderly have been reasonably well defined. Patients in the middle years (sixth and seventh decades) present a special problem in the choice of a prosthesis. To better elucidate the failure rate of the Carpentier-Edwards bioprosthesis in middle-aged patients, a comparative study of value failure rates was conducted using the Wilcoxon (Breslow) statistical technique. From September 1978 to December 1986, 502 patients underwent valve replacement with a Carpentier Edwards bioprosthesis. All patients were operated on by a single surgical team using precisely the same method of valve implantation and myocardial preservation. The overall 30-day mortality was 8.4%. PATIENT SURVIVAL: Follow-up was obtained on all 460 hospital survivors and extends to 109.2 months with a mean of 36.8 months. The cumulative survival is 1,410.6 patient-years. VALVE SURVIVAL: The 481 patients that left the hospital were divided into two subgroups. Group I included patients aged 55 to 69 years; group II, 70 years and older. There were 8 valve failures in group I. The percent of valves free of failure plotted by the actuarial method is 95.4% at 5 years (SEM 1.7, 81 valves at risk) and 95% at 7 years (SEM 1.7, 23 valves at risk). In group II (age 70 and above), there were only two valve failures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980040 TI - Comparison of Hancock I and Hancock II bioprostheses. AB - The Hancock II bioprosthesis was developed in order to provide the advantageous low pressure fixation, improved delrin stent design, and anticalcification treatment. These changes were made 6 years ago after 10 years of experience with the high pressure fixed rigid implantation ring and polypropylene stent used in the Hancock I valve. In 1983, based on our own experience with low pressure fixed valves in 76 patients, we began early clinical trials with the Hancock II valve. All valves were studied postoperatively by intraoperative catheterization and followed up with postoperative echocardiograms for measurement of valve gradients and areas. This series of 104 patients with Hancock II valves was then compared retrospectively with 119 patients receiving Hancock I valves from 1975 to 1983. A comparison of mortality, thromboembolism, and hemorrhage rates was not significantly different between groups and the valve failure incidence of Hancock I valves was an anticipated 2.34% per patient-year. There has been one primary tissue failure in the Hancock II series. This patient had fibrinous excrescences on the outflow surface of the valve in the aortic position. These nodules were compatible with an old thrombotic process of ill-defined nature. Further investigation resulted in reports of this phenomenon, which had resulted in early valve stenosis, from other centers implanting the Hancock II valve. In conclusion, the Hancock II bioprosthesis has theoretical advantages over the Hancock I in stent design, fixation pressure, and anticalcification potential. There is an unusual thrombotic process in aortic valve replacements that we have not observed in the Hancock I group or in our experience with other porcine xenografts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980041 TI - Bioprostheses at twelve years. AB - From January 1976 through December 1986, seven different types of bioprostheses have been implanted in our center. The following bioprostheses (total 1,414) were implanted in 1,098 patients: Carpentier-Edwards 567, Hancock 302, Liotta 268, Ionescu-Shiley 127, Angell-Shiley 72, Vascor 68, Implamedic 10. Follow-up ranged from 1 to 12 years, cumulative duration of follow-up was 6,747 patient-years and 8,637 valve-years, being 95.4% complete. Cumulative actuarial probability of being free from tissue valve failure (TVF) was 85.1% +/- 2.0% at 10 years, and 61.6% +/- 9.6% at 12 years. Actuarial probability of being free from TVF was 71.2% +/- 10.8% at 12 years for Carpentier-Edwards, 51% +/- 21.7% at 12 years for Hancock, 73.4% +/- 14.1% at 11 years for Angell-Shiley, 53% +/- 27.4% at 9 years for Liotta, 68% +/- 14.8% at 11 years for Ionescu-Shiley, 53.2% +/- 22.2% at 7 years for Vascor, 72.2% +/- 21.5% at 5 years for Implamedic bioprostheses. In this comparison of seven different bioprostheses, there is a large group of valves behaving in a very similar way. Only a few prostheses showed a constant and early negative trend. PMID- 2980042 TI - Long-term evaluation of the Ionescu-Shiley pericardial xenograft bioprosthesis in the aortic position. AB - From August 1977 through October 1984, 241 patients underwent aortic valve replacement with the Ionescu-Shiley pericardial valve. There were 130 males and 11 females in this series with an average age of 50.8 years (range 15 to 78 years). Isolated aortic valve replacement was performed in 121 patients (50.2%) and associated cardiac surgery in 120 (49.8%). Valve size was 21 mm or smaller in 107 cases (50.2%). Cumulative duration of follow-up was 1,260 patient-years with a mean follow-up of 5.16 years per patient. Five patients were lost to follow-up. Maximum follow-up was 10.5 years. There were 26 hospital deaths and 15 late deaths (1.19% per pt-yr). The expected 10.5 year actuarial survival rate is 82% +/- 2.9%. Twelve thromboembolic episodes occurred in seven patients--seven central and five peripheral events. The thromboembolic rate was 0.95% per patient year--32% for patients with isolated aortic valve replacement and 0.63% for patients with concomitant surgery. Freedom from thromboembolic episodes at 10.5 years is 73% +/- 12%. Structural valve deterioration was found in 24 patients (1.9% per pt-yr) with an actuarial freedom from primary tissue failure of 77.5% +/- 5.4% at 10.5 years. Reoperation was required in 39 cases (3.09% per pt-yr)- primary tissue failure (n = 24), paravalvular leak (n = 7), infective endocarditis (n = 6), and valve thrombosis (n = 2). PMID- 2980043 TI - Five-year follow-up of Hancock pericardial valves: management of premature failure. AB - The durability and function of bovine pericardial valves are dependent upon design, preservation, patient factors (age, sex), and site of valve implantation. In 1983, a shelf recall of all Hancock bovine pericardial valves (HPV) was instituted by the manufacturer. This report represents the results of an organized 5-year follow-up in a hospital Prosthetic Valve Registry of 129 HPV implanted in 122 patients (79 males, 43 females) between May 1982 and April 1985 using echo Doppler and careful clinical evaluation. Mean age was 56 +/- 15 years. There were 81 AVR, 33 MVR, 7 DVR, and 1 TVR. Concomitant coronary bypass was performed in 38 patients (31%). Surgery was on a redo basis in 25 patients (20%), urgent in 14 (11%), and for SBE in 8 patients (7%). There were seven hospital deaths (5.7%). Mean follow-up was 44 months (maximum 66 months) for 114 patients (99% complete), representing 417 patient years. There have been 20 late deaths (18%), of which 7 were directly valve related. Linearized frequency of major events (per pt-yr) was: thromboembolism, 1.6%; anticoagulant related hemorrhage, 0.8% (1 late death); prosthetic valve endocarditis 1.3%; primary tissue failure, 5.8%. Patient symptomatology was a more accurate predictor of bioprosthetic failure requiring reoperation than echo Doppler studies, which were completed in 74 of 97 patients examined during scheduled follow-up visits. Twenty-four of the 96 patients (25%) have required re-replacement at a mean interval of 44 months (27-59 months) from initial implantation. This was due to vertical shear starting at the top of the strut anchoring commissural attachments in every case.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980044 TI - Aortic valve replacement with the Carpentier-Edwards pericardial bioprosthesis: clinical and hemodynamic results. AB - From 1981 to 1987, 189 patients underwent isolated aortic valve replacement with the Carpentier-Edwards bovine pericardial bioprosthesis. There were 9 early deaths (30-day mortality of 4.8%) and 18 late deaths (3.3% pt-yr). After 5 years, the actuarial survival rate was 81.9%. All patients but one were followed for an average of 37 months after operation, and follow-up totaled 551 patient-years. Valve-related complications included 12 thromboembolic episodes (2.2%/pt-yr), 7 endocarditis (1.3%/pt-yr), 2 hemorrhages due to anticoagulation (0.4%/pt-yr), and 3 reoperations (0.5%/pt-yr). After five years, freedom from thromboembolic events was 91%, that from reoperation was 98%, and 87% for all valve-related complications. There was no primary tissue failure of the valve. Before operation, 55% of the patients were in functional Class III or IV, 97% of the survivors were in Class I or II after valve replacement. Hematological evaluation of 77 patients, an average of 18 months postoperatively, showed no clinically significant hemolysis. Postoperative hemodynamic studies in 28 patients showed mean aortic pressure gradients across the prosthesis at rest ranging from 24 mmHg with the smaller (19 mm) to 11 mmHg with the larger (25 mm and more) valves, and effective orifice areas ranging from 1.0 to 1.9 cm2, respectively. Thus, the Carpentier-Edwards pericardial bioprosthesis appears to be a reliable and hemodynamically effective substitute for replacement of the aortic valve. PMID- 2980045 TI - The mitral medical pericardial bioprosthesis: new generation bovine pericardial prosthesis. AB - The Mitral Medical Mitroflow pericardial bioprosthesis was implanted in 99 operations (99 patients with 104 valves) between 1984 and 1987. The mean age of the patients was 62.8 years (range 28 to 94 years). The early mortality was 6.1% (including patients with concomitant procedures--coronary artery bypass and ascending aortic aneurysm resection). Late mortality was 2.3% per patient-year. The total cumulative follow-up was 172 years. Thromboembolism (TE) was 4.1% per patient-year (fatal 1.2% per patient-year) (minor 2.3%; major 1.8%); antithromboembolic therapy-related hemorrhage (ATH) 0.6% (fatal 0.6%); prosthetic valve endocarditis (PVE) 0.6% (fatal 0%); periprosthetic leak (PPL) 0.6% (fatal 0%); clinical valve dysfunction (CVD) 1.2% (fatal 0%); and structural valve deterioration/primary tissue failure (SVD) 1.2% per patient-year (fatal 0% per patient-year). Thromboembolism was the significant complication. SVD occurred in two patients during the third year of evaluation. The overall patient survival was 89.2 +/- 3.3% at four years. The patients were classified as 91.9% NYHA functional Class III and IV preoperatively and 97.7% Class I and II postoperatively. The freedom at four years from TE was 88.5% +/- 4.9%; SVD 92.5 +/- 5.1%; and reoperation 89.8% +/- 5.3%. Freedom from all valve-related complications at four years was 76.8% +/- 6.6%; valve-related mortality 97.2 +/- 2.0%; mortality and reoperation (valve failure) 87.3% +/- 5.5%; mortality and residual morbidity (treatment failure) 95.5% +/- 2.6%; and mortality, residual morbidity, and reoperation (valve failure and dysfunction) 85.6% +/- 5.7%. There were three valve-related deaths of a total of ten deaths (early 2; late 1) (TE 2; ATH 1; CVD 0; PVE 0; PPL 0; and SVD 0). Valve-related reoperation was performed in four patients (TE 0; CVD 0; PVE 1; PPL 1; and SVD 2). The Mitral Medical Mitroflow pericardial bioprosthesis has provided very satisfactory clinical performance and afforded patients excellent quality of life. PMID- 2980046 TI - Pulsed, continuous, and color flow Doppler echocardiographic assessment of normal and abnormal Ionescu-Shiley pericardial valves. AB - We studied the value of pulsed, continuous, and color flow Doppler echocardiography for identifying normal and abnormal function in 51 Ionescu Shiley bovine pericardial valves. In 15 patients with clinically normal functioning mitral valves, the peak and mean pressure gradients as calculated by the modified Bernoulli equation were 8.5 +/- 4.9 mmHg (mean +/- SD) and 2.6 +/- 2.9 mmHg, respectively. The mitral valve area as calculated by the pressure half time method was 2.2 +/- 0.6 cm2. In 18 patients with normally functioning aortic valves, the peak and mean pressure gradients were 22.9 +/- 9.0 mmHg and 11.1 +/- 5.0 mmHg, respectively. In five patients with suspected mitral prosthetic dysfunction, two-dimensional and Doppler echocardiography showed: two had mitral stenosis with a mean gradient of 14.2 mmHg and mean mitral valve area of 1.0 cm2, two had severe mitral regurgitation, and one was thickened but nonstenotic. In 13 patients with suspected aortic prosthetic dysfunction, two-dimensional and Doppler echocardiography showed: two patients had markedly thickened leaflets and mean aortic gradient of 41.5 mmHg, two patients had mild aortic regurgitation, four patients had severe aortic regurgitation, and five patients had systolic murmurs secondary to pathology unrelated to the prosthetic aortic valve. Of the three patients with severe aortic regurgitation who had color Doppler, the site and direction of the regurgitant jet was correctly identified in all compared to intraoperative assessment. Pulsed, continuous, and color flow Doppler are useful as noninvasive techniques in assessing Ionescu-Shiley valve hemodynamics and function. PMID- 2980047 TI - Tissue and mechanical valves: mutually advantageous interplay. AB - This report is concerned with the dynamic interplay between glutaraldehyde preserved tissue valves (bioprostheses) and mechanical valves. These two classes of valve replacement devices are not competitive, but provide some nonoverlapping characteristic advantages and disadvantages. By proper selection, it may be possible to tailor the kind of device used for a particular patient, thus improving the overall results of bioprosthetic and mechanical valve replacement. Careful selection of patients according to age and the safety of anticoagulation should achieve a series of patients with mechanical and bioprosthetic valves that would be superior to a series in which all patients received a single device. Thus, these devices should be viewed as complimentary rather than competitive since the value of properly matching a prosthesis to the patient will be reflected in improved overall results with each class of prosthesis. PMID- 2980048 TI - Physiological manometric tested tricuspid valvuloplasty. AB - Tricuspid insufficiency associated with severe left-sided valvular heart disease carries a poor prognosis. Twenty-two patients with severe pulmonary hypertension and tricuspid insufficiency underwent a tricuspid valvuloplasty in addition to left-sided single or double valve replacement. The tricuspid valvuloplasty was performed after weaning the patient from cardiopulmonary bypass. The efficacy of the tricuspid valvuloplasty was gauged by continuous right atrial pressure recordings as the annuloplasty was completed. Obliteration of the peak of the V wave of the right atrial pressure recordings indicated that the tricuspid annuloplasty was secured. Twenty-one patients were long-term survivors, and 19 patients are in good condition for an average follow-up of 6.1 years (1 to 12 years postoperatively). PMID- 2980049 TI - Technical considerations in reoperation for porcine bioprosthetic valve failure. AB - Reoperation in porcine valve recipients is becoming increasingly frequent, owing to the limited durability of the valves. In reviewing our experience with first reoperation for porcine valve failure in 191 patients over a 17-year period, we found that following certain routine surgical steps can minimize complications and improve the surgical outcome. Extended dissection of the heart is useful during mitral bioprosthetic replacement to enhance visualization of the failing device while retracting the left atrium without undue tension on fragile structures such as the left innominate vein-superior vena cava junction. This maneuver can be avoided, however, when replacement of an aortic bioprosthesis is required. Excision of a porcine valve is performed by peeling off the fibrous tissue overgrown on the sewing ring, which exposes the underlying sutures, cutting each knot, and finding a plane between the stent and the native valve annulus by careful blunt dissection. Care is taken not to cut the Dacron cloth of the sewing ring to avoid the potential for embolization of the silicone sponge contained within it. The use of this technique helps to minimize postoperative complications such as paravalvular leak or atrioventricular block and to preserve the native annulus, which facilitates insertion of a new prosthesis. Our experience indicates that first reoperation in porcine valve recipients can be performed with low risk, particularly in elective cases, and with a low incidence of complications related to repeat median sternotomy. PMID- 2980050 TI - An improved technique for the internal mammary artery coronary bypass graft procedure. AB - An improved technique for internal mammary artery graft preparation is described. Following cautery dissection of the internal mammary artery (IMA) pedicle, the pedicle investing fascia is incised to the adventitial level along a single plane. This incision allows the tissue around the internal mammary artery to fall away and severs the muscular constrictions that often surround the internal mammary artery. Balloon calibration is performed to identify remaining constrictions and to relieve internal mammary artery spasm. A shear force limiting gauge insures that the exerted balloon force remains below the level demonstrated to cause intimal damage during electron microscopic studies. This technique allows full internal mammary artery distention without the devascularizing effects of full skeletonization. Distention of the distal internal mammary artery provides an enlarged hood to facilitate suture placement. Elongation of the internal mammary artery during balloon calibration aids in the performance of sequential grafts. This technique has been applied to 793 patients over the past five years. Postcalibration flow rates increased 3- to 18-fold over precalibration flow rates. Two early occlusions occurred during this series, one due to endothelial strippage prior to the development and use of the shear force limiting gauge. Follow-up showed 93.3% of patients to be asymptomatic. This combination of fascial incision and balloon calibration appears to offer safe technical and functional improvements to the performance of the internal mammary artery graft. PMID- 2980051 TI - A technique for concurrent repair of pectus excavatum and intracardiac defects. AB - Pectus excavatum deformity may present problems when median sternotomy is required for repair of intracardiac defects. We have used a partial median sternotomy extending from the manubrium to the level of the second intercostal space with extension into that space. The perichondral sheaths are disconnected from the sternum on the right side after removing the deformed costal cartilages. Exposure is thus provided for the cardiac procedure. Following this, the upper sternal fragments are sutured together and the repair is completed in a manner similar to the Ravitch technique. PMID- 2980052 TI - Sutureless repair of postinfarction cardiac rupture. AB - A 62-year-old female with a history of progressive angina experienced an acute myocardial infarction. Seven days later, cardiac rupture ensued. She underwent surgical repair without the aid of extracorporeal circulation. A Teflon patch was glued over the myocardial tear with medical adhesive. She recovered and is leading a normal life, 15 months after surgery. PMID- 2980054 TI - Aortic valve replacement with stentless porcine bioprostheses. AB - Aortic valve replacement with stentless porcine valve should provide superior hemodynamic results to stented porcine valve because the obstruction caused by the stent and the sewing ring is eliminated. In addition, the coronary sinuses of the recipient may allow for better dissipation of the mechanical stress to which the leaflets are subjected during diastole, thus enhancing durability of the heterograft. Aortic valve replacement with stentless glutaraldehyde-fixed aortic porcine bioprosthesis was carried out successfully in six young sheep. These animals were hemodynamically evaluated at 3 to 6 months after operation and found to have no resting gradients or any degree of aortic regurgitation. Explantation of the aortic heterograft revealed that it was well healed in the aortic root and had no evidence of any calcification. A clinical trial has been initiated and the results in the first five humans who underwent aortic valve replacement with a stentless porcine aortic bioprosthesis have been satisfactory. PMID- 2980053 TI - Long-term results after extracardiac valved conduits implanted for complex congenital heart disease. AB - Between August 1982 and December 1986, 56 patients survived implantation of an extracardiac valved conduit for complex congenital heart disease. The mean age at operation was 4.2 years (16 days to 24 yrs) and the mean weight was 15.9 kg (2.4 to 93.0 kg). The diagnosis was pulmonary atresia (PA) with ventricular septal defect (VSD) in 13 patients, tetralogy of Fallot in 11, transposition of the great arteries (TGA) with VSD in 8, truncus arteriosus, in 7, complex left ventricular outflow tract obstruction (LVOTO) in 6, complex left atrioventricular valve obstruction in 4, double outlet right ventricle with VSD and subaortic obstruction in 3, univentricular heart with pulmonary stenosis in 2, TGA with LVOTO in 1, and PA with intact ventricular septum in 1. In 35 patients, a preclotted conventional Dacron conduit (CDC) with bioprosthetic valve was used, in 19 patients a collagen-sealed Tascon valved conduit (TC) was implanted, and in 1 patient an aortic homograft was used. In a mean follow-up of 32.5 months (9 to 64 mo), there were two deaths (2/56, 3.6%) that were not related to the conduit. All survivors have been evaluated by two-dimensional and Doppler echocardiography, and 29/56 (51.8%) underwent cardiac catheterization. Nine patients (9/56, 16.1%) underwent successful valved conduit replacement, in seven cases with a nonvalved conduit. There was a significant difference (P = .011) with regard to the incidence of conduit replacement between the group with CDC (2/36, 5.5%) and the group with TC (7/19, 36.8%). Five patients underwent percutaneous transluminal balloon dilatation of the prosthetic conduit, with adequate relief of the gradient in four patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980055 TI - The Carpentier-Edwards supra-annular porcine bioprosthesis: new generation low pressure glutaraldehyde fixed prosthesis. AB - The Carpentier-Edwards supra-annular porcine bioprosthesis was implanted in 1,576 operations (1,536 patients with 1,704 valves) between 1981 and 1987. The mean age of the patients was 61.9 years (range 13 to 87 years). During the years 1981 and 1987, approximately 90% of the total valvular surgery population received the prosthesis. The early mortality was 7.0% (included patients with concomitant procedures including coronary artery bypass and ascending aortic aneurysm resection). Late mortality was 4.0% per patient-year. The total cumulative follow up was 4,237 years. Thromboembolism (TE) was 2.4% per patient-year (fatal 0.4% per patient-year) (minor 1.2%; major 1.3%); antithromboembolic therapy-related hemorrhage (ATH) 0.5% (fatal 0.07%); prosthetic valve endocarditis (PVE) 0.5% (fatal 0.2%); periprosthetic leak (PPL) 0.3% (fatal 0%); clinical valve dysfunction (CVD) 0.3% (fatal 0.02%); and structural valve deterioration/primary tissue failure (SVD) 0.2%/patient-year (fatal 0%). Thromboembolism and structural valve deterioration were the significant complications, SVD occurring primarily between the fourth and fifth year of evaluation. The overall patient survival was 77.1% +/- 1.4% at five years. The patients were classified as 89.6% NYHA functional Class III and IV preoperatively and 96.1% Class I and II postoperatively. The freedom at five years from TE was 90.6% +/- 1.0%; SVD 98.9% +/- 0.5%; and reoperation 95.9% +/- 0.9%. Freedom from all valve-related complications at five years was 84.1% +/- 1.3%; valve-related mortality 96.8% +/- 0.7%; mortality and reoperation (valve failure) 92.9% +/- 1.1%; mortality and residual morbidity (treatment failure) 93.7% +/- 0.9%; and mortality, residual morbidity, and reoperation (valve failure and dysfunction) 90.0% +/- 1.2%. There were 28 valve-related deaths of a total 280 deaths (early 4, late 24) (TE 17; ATH 3; CVD 1; PVE 7; PPL 0; and SVD 0). Valve-related reoperations were performed in 32 patients (TE 2; CVD 4; PVE 7; PPL 11; and SVD 8). The supra-annular Carpentier Edwards porcine bioprosthesis has provided very satisfactory clinical performance and afforded patients an excellent quality of life. The long-term durability of this low pressure glutaraldehyde fixed prosthesis will be determined by observation over the next five to seven years. PMID- 2980056 TI - The cross-linking and structure modification of the collagen matrix in the design of cardiovascular prosthesis. AB - Glutaraldehyde cross-linking of native or reconstituted collagen fibrils and tissues rich in collagen significantly reduces biodegradation. Other aldehydes are less efficient than glutaraldehyde in generating chemically, biologically, and thermally stable cross-links. Implants of collagenous materials cross-linked with glutaraldehyde are subject long-term to calcification, biodegradation, and low-grade immune reactions. We have attempted to overcome these problems by enhancing cross-linking through (a) bridging of activated carboxyl groups with diamines and (b) using glutaraldehyde to cross-link the epsilon-NH2 groups in collagen and the unreacted amines introduced by aliphatic dismines. This cross linking reduces tissue degradation and nearly eliminates humoral antibody induction. Covalent binding of diphosphonates, specifically 3-amino-1 hydroxypopane-1, 1-diphosphonic acid (3-APD), and to a lesser extent chondroitin sulfate to collagen or to the cross-ling-enhanced collagen network reduces its potential for calcification. Platelet aggregation also is reduced by glutaraldehyde cross-linking and nearly eliminated by the covalent binding of chondroitin sulfate to collagen. The cytotoxicity of residual glutaraldehyde can be minimized by chemical neutralization and thorough rough rinsing. PMID- 2980057 TI - Long-term fate of valve cusp patches for right ventricular outflow tract reconstruction. AB - Six patients with tetralogy of Fallot, pulmonary atresia, or absent pulmonary valve syndrome were operated with a RVOT patch containing a single aortic allograft cusp from an adult donor. The method was based on animal work with similar patches implanted in growing puppies and proved that the single cusp functioned as an effective RV outflow valve for several weeks after surgery. Postoperative angiograms in the animals confirmed nearly completely competent valves. Sacrifice of animals at 3 months documented that the allograft tissue was flexible without early deterioration or calcification. Clinical application of this method raised several questions regarding the procedure, its efficacy, and subsequent fate of the allograft valve in children requiring RVOT reconstruction. There were six children age 19 months to 10 years who required division of the pulmonary valve annulus to relieve pulmonary outflow obstruction or with absent pulmonary valve syndrome and compromised airways from pulmonary insufficiency. One patient had a positive culture from the valve and required immediate removal of the infected allograft. Patients had clinically competent pulmonary valves immediately after surgery with decreased right ventricular systolic and end diastolic pressures when compared with patients in whom nonvalve patches were inserted. All patients had some degree of pulmonary insufficiency, but no symptoms. Follow-up up of five patients available 4 to 6 years after surgery revealed no patients with stenosis of the valve cusps, although two patients have severe, one moderate, and two mild pulmonary insufficiency. We conclude that a patch containing a single adult allograft valve cusp should be considered for all patients requiring reconstruction of the RVOT, particularly if RV overload is expected to compromise the immediate postoperative result. PMID- 2980058 TI - Balloon dilatation of stenotic right ventricle-to-pulmonary artery conduits. AB - In seven children with obstructed right ventricle-to-pulmonary artery conduits, balloon dilatation (BD) was performed 10-58 months after insertion of the conduit. Average valve gradient was reduced from 69 to 32 mmHg. Obstruction at the conduit-branch pulmonary artery connection became apparent after dilatation of the valve; these distal stenoses also were dilated. It is concluded that the useful lifetime of a right ventricle (RV) to pulmonary artery (PA) conduit may be extended by BD of an obstructed biological valve and/or BD of a stenotic conduit pulmonary artery anastomosis. PMID- 2980059 TI - The biomechanics of lumbar disc herniation and the effect of overload and instability. AB - A multipart study has been performed to provide a mechanical explanation for the epidemiologic association between sitting in static (e.g., factory or office) or vibration (e.g., car or truck driving) environments and acute herniated lumbar discs. It was shown that a 1 h exposure to sitting environments caused significant changes in the mechanical properties of the lumbar intervertebral disc. During many of the latter tests, specimens were unstable (exhibited by a sudden, large flexion and/or lateral bend rotation response to an axially applied load). This showed that a motion segment in the lumbar spine could suddenly buckle and apply a tensile impact loading to the posterolateral region of the disc. We also demonstrated that a combined lateral bend, flexion, and axial rotation vibration loading could cause tracking tears proceeding from the nucleus through the posterolateral region of the anulus. It suggests that a mechanism for disc herniation is mechanical changes leading to instability of the motion segment. These experiments complete the argument that lumbar disc herniations can be a direct mechanical consequence of prolonged sitting in static or vibration environments. PMID- 2980060 TI - Lumbar disc herniations: the predictive value of the Health Attribution Test (HAT) and the Minnesota Multiphasic Personality Inventory (MMPI). AB - Ninety-one patients who were treated for lumbar disc herniation with chymopapain chemonucleolysis were evaluated preoperatively by means of the Health Attribution Test (HAT) and the Minnesota Multiphasic Personality Inventory (MMPI). There were 54 good, 10 fair, and 27 poor results after chemo-nucleolysis. Nineteen patients subsequently underwent lumbar laminectomy and discectomy and the ultimate outcome for the entire series including these laminectomy patients was 66 good, 10 fair, and 15 poor results. The fair/poor chemonucleolysis outcome patients scored significantly lower than did the good outcome patients on the HAT Powerful Others and significantly higher on the Chance scale. Patients with fair or poor outcomes after chemonucleolysis only scored significantly higher on the Hypochondriasis, Hysteria, Psychopathic Deviate, Paranoia, and Hypomania scales in preoperative MMPI testing. Good versus fair/poor ultimate outcome patients differed significantly on preoperative MMPI Hypochondriasis, Hysteria, Psychopathic Deviate, Paranoia, Psychasthenia, Schizophrenia, Hypomania, and Social Introversion scales. These groups also differed significantly on preoperative HAT Internal and Chance scales. Further analyses found the MMPI to be a slightly better predictor of chemonucleolysis outcome and much better predictor of ultimate outcome than the HAT. PMID- 2980061 TI - Fatigue fracture morphology in human lumbar motion segments. AB - The fatigue behavior of 17 fresh lumbar spine motion segments was examined during cyclic, axial, compressive loading. The loading frequency and magnitude were chosen to simulate vigorous physiological loading conditions. The average failure occurred after 318 such loading cycles. Two main types of failure occurred and both involved the endplate and the adjacent subchondral spongy bone of the vertebral body. A tendency was observed toward a relation between the degree of disk degeneration and fracture type. Location of the fatigue fractures corresponded to the location of earlier detected microcalluses of the spongy trabeculae in the vertebral body. The findings further demonstrated that, under repetitive physiological loading, the weakest part of the lumbar motion segment was the endplate and the adjacent spongy bone. PMID- 2980062 TI - Neurofibromatosis hyperkyphosis: a review of 33 patients with kyphosis of 80 degrees or greater. AB - Thirty-three patients with hyperkyphosis secondary to neurofibromatosis were reviewed as to the natural history of the deformity, complications of the untreated deformity, and results of treatment. The kyphoses ranged from 80 to 180 degrees, averaging 132 degrees. The natural history was not good, six patients presenting with paraparesis and six with respiratory distress. Twenty-seven patients had operative treatment. Laminectomy was useless for the treatment of paraparesis due to angulated spine deformity. Combined anterior and posterior spine fusion using abundant amounts of autogenous bone for both, coupled with prolonged rigid immobilization, gave the best results. Halo traction and halo casts were invaluable for the more severe deformities. Instrumentation was often impossible due to the severe deformity. All patients operated on by the authors eventually achieved solid union. PMID- 2980063 TI - Cervical spondylitic myelopathies: surgical treatment. AB - The surgical approach to the treatment of cervical spondylitic myelopathy has been one of major controversy, both clinically and in the literature. We present here the results of a retrospective analysis of 206 patients treated by the authors over a 7-year period by either a posterior or anterior approach. Our results suggest that if the procedure is tailored to the cause as well as extent of the problem, similarly good results can be obtained by either surgical approach. PMID- 2980064 TI - Osteomyelitis of the odontoid process. AB - Pyogenic osteomyelitis of the odontoid process is a rare condition requiring a high index of suspicion for diagnosis. The three cases presented illustrate that patients with severe neck pain, aggravated by rotation, and persistent fever without apparent source should be studied carefully to exclude infection of the C1-C2 area. The unusual anatomy of the C1-C2 articulation may make routine diagnostic studies difficult to interpret. Computed tomography, magnetic resonance imaging, and 111In-labeled white blood cell scans may improve diagnostic accuracy. Treatment includes rigid immobilization, high dose antibiotics, and surgical stabilization in selected cases. PMID- 2980065 TI - Cortical evoked potential monitoring during spinal surgery: sensitivity, specificity, reliability, and criteria for alarm. AB - Cortical somatosensory evoked potentials (EP) were used for intraoperative monitoring of spinal cord integrity during 158 consecutive cases involving surgical correction of spinal deformities. Using a monitoring technique designed to optimize recording, 88% of patients had stable EP results at the time of spinal manipulation. However, five of six patients with preexistent neurological deficits did not have stable EPs. A persistent EP amplitude attenuation of greater than or equal to 40% was a sensitive and specific criterion for alarm. Seven patients had transient attenuations of greater than or equal to 40% after distraction that recovered, so that the distraction instrumentation could be left in place. Overall, this particular cortical somatosensory EP recording technique appears to be appropriate for clinical monitoring. Other recording techniques may be necessary for patients with preexisting neurological deficits. PMID- 2980066 TI - Modification of the degenerative response in experimental intervertebral disk herniation in rabbits. AB - Experimental intervertebral disk degeneration is produced by ventral surgical herniation of the nucleus pulposus in rabbits. Posterior methylmethacrylate immobilization can modify the degenerative response but will only do so if applied by 2 days after ventral herniation. PMID- 2980067 TI - Intraoperative ultrasonography (IOUS) in thoracolumbar fractures. AB - The thoracolumbar levels are the second most common region for spinal trauma. A major surgical effort often entails removal of retropulsed bone fragments with decompression of the spinal contents or realignment of vertebral subluxations. The ability to determine intraoperatively the completeness of such a procedure could impact on the surgical approach and, ultimately, the operative result. The intraoperative use of ultrasonography has gained popularity and applicability. This comparison study of intraoperative ultrasonography versus postoperative computed tomography (CT) assessed the accuracy of intraoperative ultrasonography in determining the status of the spinal canal following surgical intervention in a group of 21 patients with thoracolumbar fractures. In all cases a patent ventral subarachnoid space or complete spinal canal decompression was deduced following intraoperative ultrasonography. The postoperative assessment by CT concurred in 20 of 21 (95%) situations. Intraoperative ultrasonography proved useful during the operative management of these fractures and gave good supportive evidence that the neural elements were decompressed by surgical procedure. PMID- 2980068 TI - The biomechanical and histomorphometric properties of anterior lumbar fusions: a canine model. AB - An in vivo model was developed to compare the biomechanical stability, incidence of radiographic union, bone formation rate, and bone graft remodeling parameters of anterior interbody fusions. Eighteen 1-year-old beagles underwent anterior and posterior spinal destabilization procedures at L5-L6 to produce a reproducible amount of spinal instability--resection of the anterior longitudinal ligament, L5 L6 intervertebral disk, L5 and L6 lamina, spinous processes, zygoopophyseal joints, and ligamentum flavum. Group I (N = 6) were surgically destabilized controls; Group II (N = 6) underwent anterior L5-L6 interbody fusion with iliac crest bone graft; and Group III (N = 6) underwent anterior stabilization with a longitudinal fibular strut graft in addition to the same operative procedure as Group II. Six months postoperatively the group with the highest incidence of successful radiographic L5-L6 arthrodesis was Group III, anterior interbody fusion and fibular stabilization (p less than .10). The rank order of biomechanical stability was the same for the three groups for both torsional and axial compressive stiffness, with Group I (destabilized controls) being the least rigid, then Group II (anterior fusion with iliac crest grafting only), and the most rigid to both torsion and axial compressive loading was Group III (anterior fusion with fibular stabilization and iliac crest bone graft). The bone formation rate [mm3/(mm3 x year) x 10(3)], which was derived from the distance between fluorochrome markers, revealed that the more stable the individual spinal construct, the lower the bone formation rate. In summary, the beagle provided a successful model for studying in vivo the response of anterior bone grafts over a 6-month interval and provided comparative biomechanical and histomorphometric data on spinal interbody fusion techniques. PMID- 2980069 TI - Magnetic resonance imaging in the diagnosis of lower thoracic disc herniation. AB - The diagnosis of thoracic disc herniation can be difficult due to the lack of a characteristic clinical presentation. In six recent cases, magnetic resonance imaging (MRI) provided excellent noninvasive definition of the pathology and its level, in spite of atypical clinical presentation, and also provided anatomic information allowing surgical planning. Two examples are presented in which myelography was not helpful, but MRI was diagnostic. MRI scanning is the radiographic procedure of choice when thoracic disc herniation is in the differential diagnosis. PMID- 2980070 TI - Epidural anesthesia for lumbar spine surgery. AB - A retrospective review of the hospital records of 80 patients undergoing elective lumbar spine surgery was performed, in order to determine the effect of anesthetic technique on various clinical parameters. Forty patients receiving epidural bupivacaine anesthesia were matched with 40 patients receiving general endotracheal anesthesia; these two groups were homogeneous based on age, sex, type of operative procedure, and number of spinal levels operated upon. Significant results included lower injectable narcotic requirements (p less than 0.001), lower incidence of postoperative urinary retention (p less than 0.01), and lower operative blood loss (p less than 0.1) for patients receiving epidural anesthesia. Epidural bupivacaine provided satisfactory anesthesia, and allowed intraoperative testing of lower extremity motor function. In a follow-up surgery, 38 of 40 patients who received epidural anesthesia were satisfied with the technique. For patients undergoing decompressive lumbar spine surgery, epidural bupivacaine anesthesia is an effective, well tolerated technique with several potential advantages, and an acceptable incidence of complications, as compared with general endotracheal anesthesia. PMID- 2980071 TI - Magnetic resonance imaging (MRI) of thoracolumbar fractures. AB - Many patients suffer neurologic deficits as a consequence of thoracolumbar fractures. A major surgical effort often entails decompression of the neural elements. Which patients would benefit from this endeavor remains to be determined. Magnetic resonance imaging (MRI) has emerged as one of the foremost neurodiagnostic procedures. This study assessed the impact of MRI on the management of these injuries in 21 patients. Five anatomical parameters were evaluated. Significant associations were noted between the neurologic morbidity from the injury and (a) the relative level of the conus medullaris and; (b) the patency of the ventral subarachnoid space. On the basis of these data, a treatment protocol is presented. MRI has gained prominence as an investigative tool in cases of thoracolumbar trauma. PMID- 2980072 TI - A comparison of Harrington rod fixation with and without segmental wires for unstable thoracolumbar injuries. AB - Between January 1, 1975 and November 1, 1986, 77 patients with acute unstable thoracic or lumbar spine fractures underwent reduction, posterior stabilization with dual Harrington distraction rods, and fusion with autogenous iliac crest bone graft at Vanderbilt University Medical Center. Beginning March 1985, in 25 patients, segmental interspinous wires were employed, in addition to the Harrington rods, to augment the surgical construct. Clinical and radiographic analysis was performed to determine if differences existed between the two groups. Average time to surgery, hospital stay, and time to brace discontinuance were lower in the segmental wire group. Pain and work status at final follow-up were similar in the two groups. There was a higher incidence of superior and inferior hook migration and reoperation for this complication in patients treated with Harrington rods alone. There was no significant difference in postoperative correction of deformity, although at follow-up, there was greater recurrence of deformity in the Harrington rod group. In summary, the use of segmental wires to supplement Harrington rod fixation appears to offer advantages over Harrington rods alone, with minimal or no increased risk. PMID- 2980073 TI - Somatosensory influence on corticomotor evoked potentials. AB - The clinical utility of corticomotor evoked potentials (CMEPs) as a method of evaluating and monitoring patients with spinal cord disorders is being intensively studied. Relatively few neuronal mechanisms responsible for waveform production are clearly known. Although CMEP components are dependent upon activity carried in descending motor pathways, somatosensory information can influence the basic waveform structure. By stimulating peripheral afferent fibers at varying frequencies, intensities, and trains, two CMEP component groups were identified based on latency. The configuration of the short-latency waveforms was influenced primarily by large-diameter afferents. Long-latency waveforms were altered primarily by small-diameter afferents. The present investigation describes both segmental and suprasegmental modification of CMEP characteristics based on afferent fiber group stimulation. If both motor and sensory systems can be accurately assessed, the clinical applications of CMEPs are considerably enhanced. PMID- 2980074 TI - Hematogenous osteomyelitis complicating a closed compression fracture of the spine. AB - A case of hematogenous osteomyelitis of a vertebral body following a closed compression fracture is presented. Staphylococcus septicemia developed 7 days after the injury. In spite of prompt antibiotic treatment, osteomyelitis of the compressed T12 was recognized 5 weeks later. The role of indium white blood cell (WBC) scanning in establishing the diagnosis is highlighted. PMID- 2980075 TI - Lumbar burst fracture. PMID- 2980076 TI - Transfusion and bone marrow transplantation. AB - Bone marrow transplantation has prolonged the lives of a significant percentage of patients with a variety of both malignant and nonmalignant disorders. However, the impact of this treatment on a transfusion service is substantial. Large numbers of often specialized blood products are required to support these patients, and the logistics of accomplishing this taxes the ingenuity and resources of even large regional blood programs. PMID- 2980077 TI - Transfusion associated AIDS. PMID- 2980078 TI - Red cell membrane changes during storage. AB - A great deal is known about the red cell membrane and its abnormalities in various pathologic states. During red cell storage there is a progressive development of spheroechniocytosis with eventual production of irreversibly nondeformable red cells. The loss of membrane function is most likely related to some abnormality in maintenance of the cytoskeleton of the red cell. These changes appear to occur independent of ATP levels. Despite the increasing knowledge of the structure and function of the red cell membrane very little as yet is known about the specific abnormality in the red cell membrane that occurs during storage in the blood bank. Recent evidence for abnormal spectrin-actin interaction and abnormal spectrin oxidation has been the most promising. Areas of interest for research include studies of the specific mechanisms by which the plasticizer DEHP interacts with the membrane, specific definition of the molecular defect in membrane proteins that occurs during storage, and means to prevent these. If such deterioration and membrane stiffening could be prevented then the quality of the red cells that are transfused would be improved both in their function and ability to survive in the microcirculation. A final need, while not of specific value to the red cell itself, is the development of media and additives that will allow for increased plasma and Factor VIII yields, one of the driving forces in the blood transfusion system. PMID- 2980079 TI - The use of therapeutic plasmapheresis for neurological disorders. National Institutes of Health Consensus Development Conference. PMID- 2980080 TI - Directed donations for pediatric patients. PMID- 2980081 TI - Rationale for surrogate testing to detect non-A, non-B hepatitis. AB - NANB hepatitis was initially recognized in 1975 and 13 years later, the exact etiology of this presumed viral disease remains uncertain. The acute illness is relatively mild with only about 25% of patients becoming icteric. Nevertheless, at least one half of the patients have evidence of chronic infection, and, as recently recognized, 10% to 20% develop severe liver disease. Because approximately 2% of patients who receive transfusions and whose underlying medical condition permits long term follow-up develop posttransfusion hepatitis, procedures for reducing this risk are considered prudent. Unfortunately specific tests for detecting NANB hepatitis are not available, and it is unlikely that such tests will be available in the near future. Hence, testing by surrogate or nonspecific tests (ALT and anti-HBc) were recommended because evidence from two studies conducted during the 1970s showed these tests identify some donors thought to transmit the infection. However, randomized, controlled prospective studies to determine whether these tests will, in fact, reduce NANB posttransfusion hepatitis were not performed. By the mid-1980s it was apparent these studies would not be performed nor were studies to determine the incidence of NANB posttransfusion hepatitis in the post-AIDS screening era likely to be initiated. Therefore, surrogate testing, as the best available method for reducing posttransfusion hepatitis, was implemented in the United States in 1986 87. PMID- 2980082 TI - Citrate toxicity during massive blood transfusion. PMID- 2980083 TI - Clinical use of blood transfusion devices. PMID- 2980084 TI - The uses of radiolabeled red cells in transfusion medicine. AB - The ability to radiolabel and follow the red cell in vivo has been of great value to the clinician and the investigator. An understanding of the advantages and disadvantages of the three nuclides used in transfusion medicine--51Cr, 99mTc, and 111In--allows the selection of the appropriate nuclide for the task at hand. For red cell life-span studies and for red cell survivals extending over one hour, 51Cr is the label of choice. The most accurate method for determining the zero-time (100% survival) point for a life span or survival study is to ascertain the RCM of the subject using autologous red cells labeled with another nuclide, preferably 99mTc. RCM studies done independently from a 51Cr survival should be performed using 111In labeled red cells. Red cell survivals are most often used to determine transfusion compatibility or to evaluate blood storage systems. There are important differences in the conduct and interpretation of red cell survivals for these two purposes. Although other promising methods for evaluating the in vivo survival of red cells are under investigation, it is likely that the radiotracers now available will continue to play a primary role in performing these studies in the future. PMID- 2980085 TI - Management of beta-thalassemia major using transfusions and iron chelation with deferoxamine. PMID- 2980087 TI - Symposium on transfusion-associated infections and immune response. PMID- 2980086 TI - Red cell polyagglutination. PMID- 2980088 TI - Transfusion-induced immunologic unresponsiveness. PMID- 2980089 TI - Transfusion-associated hepatitis B: past and present. AB - Thus, it can be concluded that: (1) Hepatitis B, even before the introduction of donor screening for HBsAg, probably never accounted for more than one-quarter of all cases of transfusion-associated hepatitis. (2) As of 1988, the frequency of this viral infection in blood recipients, now that sensitive donor screening for HBsAg is routine, is less than 10%. The exception are multiply transfused populations, such as hemophiliacs, the majority of whom have evidence of current or past exposure to hepatitis B. However, new inactivation procedures are likely to reduce the attack rate in virgin populations of hemophiliacs. (3) Interdiction of paid blood has proved far more effective than donor screening for HBsAg in reducing the overall frequency of transfusion-associated hepatitis. (4) The current requirement that all donors be screened for anti-HBc and ALT as surrogate markers of non-A, non-B hepatitis infection, and for anti-HIV, is likely to reduce the hepatitis B attack rate even more, perhaps to near zero. (5) The long term outcome of transfusion-associated hepatitis B has not been determined, a task that will remain difficult to accomplish because of the paucity of current cases. Information in this regard will need to be derived from the recall and reevaluation of pedigreed patients who participated in prospective studies that were conducted in the distant past. PMID- 2980090 TI - Biology of hepatitis delta virus. PMID- 2980091 TI - Transfusion-related delta hepatitis. PMID- 2980092 TI - An hypothesis on the cost effectiveness of homologous blood transfusions in 1988. PMID- 2980093 TI - Laboratory diagnosis of HIV infection. PMID- 2980094 TI - Molecular analysis of hepatitis B virus infection. PMID- 2980095 TI - Science and technology in mitigating AIDS. AB - The workshop emphasized contributions of genetic engineering in providing reagents and techniques for diagnosing and monitoring HIV infections. Despite a variety of tests for specific antibodies, virus antigens and nucleic acids no consistent serum profile has emerged that correlates well with virus life cycle or clinical course. HIV infections are a great deal more complex than hepatitis B infections where diagnosis and prognosis are very accurately based on serologic profiles. HIV generates at least 15 virus proteins all of which are immunogenic. The workshop emphasized studies on immunogenicity of the proteins. Full understanding of the virus life cycle and the clinical course of disease may require in-depth studies of the production and immunogenicity of the virus directed catalytic and regulatory gene products. Fortunately, all of these reagents can be produced through biotechnology. The new techniques described in the workshop will allow expanded studies to proceed rapidly. PMID- 2980096 TI - Research in progress workshop. PMID- 2980097 TI - Morphometric evaluation of capillary basement membrane thickness in the quadriceps muscle of diabetic and nondiabetic Chinese hamsters. AB - Quadriceps muscle capillaries from 19-23 month old genetically diabetic (XA and AC) and nondiabetic (M) subline Chinese hamsters were morphometrically evaluated to determine if capillary basement membrane thickening (CBMT) is a quantifiable complication of diabetes. Significant CBMT was present in the diabetic XA Chinese hamsters (49.37 nm +/- 17.81, p less than 0.007) in comparison with the nondiabetic M hamsters (34.08 nm +/- 9.98). Although there was a trend towards expansion of the muscle capillary basement membranes in the diabetic AC Chinese hamsters, the value was not statistically significant. A nested analysis of variance showed that the greatest source of variation in basement membrane thickness occurred among capillaries within each animal. In addition, a positive correlation (r = 0.62; p less than 0.002) existed between blood glucose levels and CBMT in the XA subline. These data should serve as guidelines for evaluation of antimicrovascular disease compounds which will be tested to determine if they prevent or retard microangiopathy in the diabetic Chinese hamster. PMID- 2980098 TI - Early embryonic development in the rat following in utero exposure to alcohol and caffeine. AB - The influence of both alcohol and caffeine on early embryonic development was investigated in pregnant rats. Compared to the corresponding controls, a high incidence of resorptions and abnormal embryos was induced following treatment of the animals with alcohol (0.015 ml/g body weight, 12.5% v/v, i.p.) on gestational days 6 through 12 and with caffeine (25 mg/kg body weight, i.v.) on gestational day 10. In addition, embryonic growth was severely affected. Reduction of placental blood circulation and impairment of cellular proliferation may account for the observed deleterious effects on the embryo. PMID- 2980099 TI - Postnatal development of dendritic reticulum cells and their immune complex trapping ability. AB - The postnatal development of dendritic reticulum cells in the rat popliteal lymph nodes was electron microscopically investigated in relation to the appearance of immune complex trapping capacity. The popliteal lymph nodes of neonatal rat consisted of loosely arranged fibroblastic reticulum cells. In the following stage, the peripheral cortex and paracortex became distinguishable. The former was made up of an accumulation of small lymphocytes, scattered within a framework of reticulum cells. On te 28 th day, the first primary follicle appeared in the peripheral cortex. Simultaneously the immune complex could be trapped on the cytoplasmic membrane of reticulum cells, which were located in the central portion of the primary follicles. The early image of germinal centers appeared corresponding to immune complex trapping areas. In the well-developed secondary follicles, the immune complex trapping cells were mainly localized in the cap area. Their cytoplasmic membranes formed the dendritic processes, on which the distinct ability of trapping of the immune complex was recognized. It was demonstrated that the fibroblastic reticulum cells, forming the stroma of lymph nodes, were transformed into the typical dendritic reticulum cells with labyrinth structures in the cap area. Desmosomal junctions were often found, not only between the dendritic reticulum cells themselves, but also between the dendritic reticulum cells and lymphocytes. We suggest that the desmosomal junctions play a role as the channel for a transmission of immunological information. PMID- 2980100 TI - Close association of centroacinar/ductular and insular cells in the rat pancreas. AB - Close contacts between endocrine insular cells and exocrine acinar, centroacinar and ductular cells occur frequently in the rat pancreas as seen by both light and electron microscopy. Islets of Langerhans are surrounded incompletely by a thin connective tissue capsule or mantle but numerous exocrine-endocrine cell contacts occur at the periphery, which is irregular with considerable "intermingling" of the two cell types. Centroacinar and ductular cells are seen to be in contact with all endocrine cell types but most commonly insulin-secreting B-cells. The basal surface of centroacinar cells in the region of contact may be extensive, sometimes with overlap of basal processes of these cells and their lateral extension between acinar and insular cells. The areas of contact contain no connective tissue or basal lamina and show no surface specializations. The presence of both the "open" and "closed" type of enteroendocrine cells within acini is confirmed, some also being in contact with centroacinar cells. The functional significance of these exo-endocrine cell contacts is discussed in terms of the endocrine-acinar portal system, possible direct paracrine secretion, compartmentalization within the islet, and the known effects of islet hormones on exocrine secretion. Also relevant is the developmental origin of islets from ductal tissue and the cellular origin of some tumours, e.g., insulinomas, from duct cells. PMID- 2980101 TI - Ultrastructure of pancreatic exocrine cells of the rat during starvation. AB - Ultrastructural changes of the pancreatic exocrine cells after 3, 7, 14, 21, 28, 35 and 42 days of starvation were observed in male rats aged from 16 to 18 months weighing between 600 and 700 grams. The number of zymogen granules after starvation decreased to less than about 70 per cent of that of the control. Changes in the rough endoplasmic reticulum were hardly seen up to 14 days of starvation as compared with the control, but were observed in the apical and basal cytoplasm of the cell from 21 days after starvation. Particularly in 35- and 42-day starved rats, the rough endoplasmic reticulum was frequently shortened and dilated, and changed to disorganized membranous structures. The lysosomes in the apical cytoplasm of the cell gradually increased in number after starvation, and contact or fusion between the zymogen granules and lysosomes (viz, so-called crinophagy) was often seen at 35 and 42 days of starvation. Large autolysosomes especially those containing zymogen granules and rough endoplasmic reticulum were also marked in the basal cytoplasm of the cell after 35 and 42 days of starvation. Alterations in the basal cytoplasm of the cell appeared later than those in the apical cytoplasm. It was considered that, owing to its role in protein synthesis, the basal cytoplasm of the pancreatic exocrine cells in starved rats might be protected as far as possible during long-term starvation. PMID- 2980102 TI - Encephalopathy with astrocytic residual bodies. Report of a case and review of the literature. AB - Biopsy and autopsy findings in a girl who died at 7 1/2 months after having suffered from progressive axial hypotonia, myoclonus, EEG changes and retarded psychomotor development. Inclusions consisting of lamellar profiles, situated in membrane-bound cytosomes were found mainly in astrocytes, but also in neurones and in axons of peripheral nerves. Lipofuscin bodies were also increased in number. The patient belongs in the same category as cases studied by Towfighi et al. (1975) and Martin et al. (1977). Etiology and pathogenesis of this syndrome remain unknown. It is suggested, however, that the pathological changes observed might have been caused by the administration soon after birth of anti-epileptic drugs (diphenylhydantoin, clonazepam and nitrazepam). PMID- 2980103 TI - Experimental study of WGA binding on the endothelial cell surface in cerebral ischemia. AB - The relationship between the saccharide chain on the endothelial cell surface and the permeability of intracerebral blood vessels has been studied. In the present study, wheat germ agglutinin (WGA) was perfused into capillaries in the area postrema of the normal Mongolian gerbil, where the blood brain barrier (BBB) is known to lack, and into intracerebral blood vessels, the BBB of which had been destroyed by experimentally induced brain ischemia. The light microscopic features of the sections from WGA-perfused brain tissues of the normal gerbil revealed that most of the blood vessels, including capillaries in the brain parenchyma, showed positive findings (the reaction induced a very distinct staining of the vascular wall) from which the course and structure of the fine vessels could be determined. The reaction to WGA on the diaphragma fenestra (DF) in capillaries in the area postrema was relatively weak, and DF without the reaction were occasionally revealed by electron microscopy. The gerbil, in which cerebral ischemia had been induced, also showed partial defect of the reaction with WGA on the luminal side of the endothelial cells. The results of the present experiment suggest some degree of correlation between the saccharide chains, including the specific monosaccharide of WGA, on the endothelial cell surface and permeability. It was considered that lectin can be used as an index for morphological observations, suggesting an alteration in function of the endothelial cell membrane. In addition, the perfusion method in this experiment suggested the possibility of distinguishing pinocytotic vesicles from pits of cell membranes. PMID- 2980104 TI - Oxygen toxicity in the infant rhesus monkey lung. Light microscopic and ultrastructural studies. AB - Eight monkeys were anesthetized with ketamine hydrochloride and positive pressure ventilated with greater than 95% oxygen (tests) or room air (controls) for 24 hours. Two test monkeys and one control were treated with E. coli endotoxin (500 micrograms/kg) IV at the start of the study and after 12 hours. Histopathological changes in the lung parenchyma were evaluated using light and electron microscopy. Interstitial edema was detected as early as 24 hours after the onset of hyperoxia but there was no significant increase in the alveolar-capillary distance (blood-air barrier). Morphologic signs of oxygen toxicity further included swelling and disruption of vascular endothelium, and swelling of alveolar Type II pneumocytes. There was no difference in the number of macrophages per high power field between the four groups but significant differences were observed in the number of neuroepithelial bodies (NEBs) per cm2 and mast cells per high power field at the light microscopic level. Treatment with endotoxin did not protect against oxygen toxicity and was associated with an exacerbation of the morphological alterations in the lung parenchyma and swelling of alveolar Type I pneumocytes. PMID- 2980105 TI - Peritoneal fine structure of inguinal hernia: a scanning electron microscope study. AB - Mesothelial cells of the normal human peritoneum of the anterior abdominal wall are covered with numerous surface microvilli. These cells become partially denuded inside the sacs of direct and indirect inguinal hernias and so lose the protective property the microvillar covering may impart on them. These mesothelial cells of hernial sacs also acquire an extensive surface coat of fibrin-like material, presumably due to the loss of that protective property, which may as a result subject them to adhesions. There is a considerable collagen build-up in the subserosal fibrous tissue of sacs of both direct and indirect inguinal hernias. Such a build-up is at variance with the accepted current surgical concept which suggests a defect in collagen synthesis, rather than a build-up, as the cause of direct hernia. PMID- 2980106 TI - Immunohistochemical localization of prolactin in functioning and regressing corpus luteum of pituitary autotransplanted rats. AB - In an attempt to shed light on the intimate mechanism by which prolactin (PRL) switches from supporting corpus luteum (CL) progesterone secretion (P) to promote structural regression of the CL, day 2 (metestrous) autopituitary transplanted (APTr) rats were used. In APTr rats the CL is under the only control of PRL since an almost complete absence of LH and FSH exist. The experimental group was given bromocriptine (CB-154: 0.4 mg/day) on days 12, 13 and 14 of the cycle and 0.25 ml of ethanol from day 15 to day 21. The control group was given CB-154 from day 12 to day 21. Rats were hemiovariectomized on day 12 to assess the morphological characteristics of the active CL. PRL and P were determined by RIA on days 12, 15 and 22. On day 12, both PRL and P levels were higher than 80 ng/ml (luteotrophic action of PRL). On day 15, due to treatment with CB-154, the levels of both hormones had fallen below 7 ng/ml (functional luteolysis). On day 22, PRL levels were again high (greater than 50 ng/ml) in the shortly CB-154-treated rats and low (less than 5 ng/ml) in the controls; the P levels were lower than 5 ng/ml in both groups. PRL-induced structural luteolysis in the experimental group (hyperprolactinemic) was assessed by the structural characteristics and by the CL weight loss on day 22 in comparison with that exhibited by control rats. The immunohistochemical staining of both endogenous and total PRL in the lutein cells showed that the internalization of PRL is not modified by the functional state of the CL, nevertheless the intracellular redistribution of the internalized hormone varied in relation with the PRL action on the CL (luteotrophic, day 12 vs luteolytic, day 22). These results seem to indicate that intracellular mechanisms rather than receptor content determine CL response to PRL. PMID- 2980107 TI - Ultrastructure of striated muscle fibers in the middle third of the human esophagus. AB - Striated muscle fibers and their spatial relationship to smooth muscle cells have been studied in the middle third of human esophagus. Biopsies were obtained from 3 patients during surgery. In both the circular and longitudinal layers, the muscle coat of this transition zone was composed of fascicles of uniform dimension (100-200 microns of diameter); some of these bundles were made up of striated muscle fibers, others were pure bundles of smooth muscle cells and some were of the mixed type. Striated muscle fibers represented three different types, which were considered as intermediate, with certain structural features characteristic of the fast fiber type. Of these, the most frequently-found fibers were most similar to the fast fiber type. Satellite cells were numerous; in mixed fascicles they were gradually replaced by smooth muscle cells. The gap between striated muscle fiber and smooth muscle cells was more than 200 nm wide. It contained the respective basal laminae and a delicate layer of amorphous connective tissue. No specialized junctions were formed between consecutive striated muscle fibers, or between striated muscle fibers and smooth muscle cells. Interstitial cells of Cajal were never situated as close to striated muscle fibers as to smooth muscle cells. PMID- 2980108 TI - The development of the neurons of the glossopharyngeal (IX) and vagal (X) sensory ganglia in chick embryos. AB - The timetable of cell generation, neuronal death and neuron numbers in the fused proximal glossopharyngeal (IX) and vagal (X) ganglion and distal IX and X ganglia were studied in normal and nerve growth factor (NGF) treated chick embryos. 3H thymidine was injected between the 3rd and 7th days of incubation and embryos sacrificed on the 11th day. Neurons in the distal IX and X ganglia were generated between the 2nd and 5th days of incubation, the peak mitotic activity occurring on the 4th and 3rd days, respectively. Neurons of the proximal IX and X ganglion were generated between the 4th and 7th days, with maximum neuron generation on the 5th day of incubation. Counts of neurons in the 3 ganglia between the 5th and 18th days of incubation showed a maximum of 22,000 on the 8th day in the proximal IX and X ganglion and this decreased to 12,000 by the 13th day. In the distal IX ganglion, the neuron number decreased by 44% from 4,500 on the 6th day to 2,500 by the 11th day. A similar decrease of 43% was found in the distal X ganglion, the neuron number falling from 11,500 on the 7th day to 6,500 by the 11th day of incubation. Neuronal cell death in these ganglia extended from the 5th to the 12th day of incubation, maximum cell death occurring at or after the cessation of mitotic activity. NGF administration from the 5th to the 11th day of incubation did not have a measurable effect on the neurons of proximal IX and X and distal IX ganglia, but increased neuronal survival by 30% in the distal X ganglion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980109 TI - Ultrastructure of the renal corpuscle of Testudo graeca (Chelonia). A comparison between hibernating and non-hibernating animals. AB - The renal corpuscle of hibernating and non-hibernating Testudo graeca was studied by means of light and electron microscopy. Renal corpuscles are small and have a glomerular architecture similar to that found in other vertebrates with a limited glomerular filtration rate. In hibernating animals, unlike non-hibernating, some morphological changes took place. The cells of the renal corpuscle were densely packed, podocytes and parietal cells showed a marked cytoplasmic vacuolization, there was a highly developed capillary basement membrane and the endothelial and mesangial cells showed abundant dense granules. These morphological features apparently correspond to a vacuolar degeneration. They may also be the morphological basis of the decrease in the glomerular filtration rate observed during this period. PMID- 2980110 TI - Atrial natriuretic peptide in the heart and pancreas. AB - We used antisera to pure atrial natriuretic peptide to localise this peptide by immunocytochemistry in rat and human tissue. We showed that both rat and human atrial cardiocytes gave a positive reaction while ventricular cardiocytes were consistently negative. Peripheral islet cells in rat but not in human pancreas also showed positive staining for ANP. We showed by double labelling techniques that the ANP was present in the glucagon containing cells. PMID- 2980111 TI - An autoradiographic study of the uptake of tritiated proline by osteoblasts during hibernation. AB - Twenty-four LSH and LVG strain golden hamsters, Mesocricetus auratus, were used. Experimental animals were maintained at 5 C and allowed to hibernate. Control animals were kept at 27 C. Six animals (3 experimental, 3 control) were injected subcutaneously with 1 microCi of 3H-proline/gm body wt. (Spec. act. 3 Ci/mM) after hibernation lasting 12 hours, 1 day, 3 days, or 7 days. Animals were killed 1 hour after injection and autoradiographs were prepared from 5 microns thick decalcified sections of femurs. A greater number of endosteal cells were labeled than periosteal cells and also exhibited a greater magnitude of labeling throughout the study. Differences between endosteal and periosteal cells both in percentage of cells labeled and magnitude of labeling were maximum in control animals and progressively decreased with increasing periods of hibernation. A reduction in synthesis of matrix proteins during the early period of hibernation was seen and was attributed to a significant reduction both in average cell activity and in the number of active cells during hibernation. The latter phenomenon apparently made a large contribution to the reduced matrical synthesis. 3H-proline uptake by osteoblasts probably reflects the reduced requirements of matrical synthesis during hibernation. PMID- 2980112 TI - Gastric cancer preceded by severe dysplasia. AB - For elucidation of histogenesis of gastric cancer derived from relatively flat mucosa, 77 cases of surgically resected stomachs with lesions of severe dysplasia in cancerous mucosae or with cancerous changes in severely dysplastic mucosae were detected out of 380 recently examined cases of early gastric cancer. Several examples of early gastric cancer of the superficial type, showing histological changes indicating that they had developed on the preexisted dysplastic mucosal lesions, have been presented, together with the background data. The frequency for detecting such changes was higher in the slightly elevated lesions than in the depressed or eroded ones. PMID- 2980113 TI - HBsAg, HBcAg and delta-Ag in liver tissue: simultaneous visualization in a single tissue section by triple immunostaining. AB - The distribution pattern of HBsAg, HBcAg and Delta-Ag was investigated by immunohistochemistry in a series of paraffin embedded liver tissue specimens from 45 subjects with serum HBsAg and anti-Delta antibody positivity. An indirect immunoperoxidase technique was used. Stains for HBsAg, HBcAg and Delta-Ag were alternatively carried out on serial tissue sections and, for the first time, consecutively in a single section (triple immunostaining). Simultaneous presence of all antigens occurred in 7 out of 45 cases, and of two antigens (HBsAg and Delta-Ag) in the remaining 38. Two antigens (either HBsAg and HBcAg or HBcAg and Delta-Ag) could also be shown in the same cell. A series of new observations was forwarded by the successful application of triple immunostaining in the present series: 1) high frequence of simultaneous presence of HBcAg and Delta-Ag (7 out of 45 cases = 16%); 2) cytoplasmic localization of Delta-Ag; 3) localization of HBcAg on the cell membrane of HBsAg positive Ground-Glass (G-G) hepatocytes; 4) Ground-Glass appearance of hepatocytic cytoplasm associated with exclusive content of HBcAg (HBcAg-Ground-Glass: a new variant of G-G-hepatocytes). PMID- 2980114 TI - Morphological differentiation of mitochondria in the early chick embryo: a stereological analysis. AB - The morphological evolution of mitochondria in three cell types of chick embryo in neurulation was analyzed by stereological methods. Mitochondria, showing a random distribution, were characterized by moderate electron-dense matrices and normal cristae. The numerical density of mitochondria significantly increased in the neuroectoderm and epiblastic cells while their volume density remained unchanged. The mitochondria in mesoderm cells were ellipsoidal (axial ratio 2:1) at stages 5 and 8 although they underwent an elongation in neuroectoderm and epiblastic cells (axial ratio from 2:1 to 1.6:1). The individual size of "average mitochondria" in the mesoderm cells was smaller than in other cell types. The total V/S (volume/surface) ratio of mitochondria decreased during neurulation. These morphological changes have been discussed emphasizing the possible metabolical role of mitochondria during morphogenesis. PMID- 2980115 TI - The kinetic differences between sodium nitrite, amyl nitrite and nitroglycerin oxidation of hemoglobin. AB - The effect of sodium nitrite, amyl nitrite and nitroglycerin (glyceryl trinitrate) on the hemoglobin of adult erythrocytes was examined in vitro. Both amyl nitrite and nitroglycerin reacted immediately with oxyhemoglobin to effect oxidation into methemoglobin while sodium nitrite required an inductionary period (lag phase) prior to the reaction. Kinetic studies of the biomolecular rate law for each of the preceding reaction's reactionary periods (log phases) allowed rate constant calculations to be made. The values are 1.14 x 10(4) M-1 min-1, 7.45 x 10(4) M-1 min-1, and 3.50 x 10(1) M-1 min-1 for sodium nitrite, amyl nitrite and nitroglycerin, respectively. A comparison of the amyl nitrite and nitroglycerin rate constants reveals that amyl nitrite is approximately 2000-fold more toxic to oxyhemoglobin than nitroglycerin. These oxidant's effect on in vitro hemoglobin solutions are comparable since both reactions approximate to rectangular hyperbolae. Sodium nitrite reacts about 300-fold faster with oxyhemoglobin than does nitroglycerin. However, the sodium nitrite reaction proceeds in a sigmoidal fashion which makes a strict comparison between these compounds relative toxicities less clear cut. PMID- 2980116 TI - Lectin (UEA-1) reaction of capillary endothelium with reference to permeability in autopsied cases of cerebral infarction. AB - The relationship between endothelial reactivity to Ulex europaeus agglutinin-1 (UEA-1) and the permeability of the vascular wall in human autopsied cases of cerebral infarction was studied. Sections from the cerebral cortex were reacted with horseradish peroxidase UEA-1 to demonstrate the surface membrane of endothelial cells. Albumin in the neuropil of sections was demonstrated for the estimation of increased vascular permeability. The results showed that endothelial reactivity to UEA-1 was reduced in cases where death had occurred 3 to 5 days after onset of cerebral infarction. Reactivity was also diminished in cases where death had occurred after 13 and 25 days; these cases showed fresh ischemic lesions caused by re-attacks of infarction. Albumin extravasation into the neuropil was demonstrated in these intermediate cases. Chronic cases, dying after more than 52 days, showed no reduction of endothelial reactivity to UEA-1 and no albumin extravasation was proved. It was concluded that UEA-1 can be employed as a useful morphological marker for evaluation of endothelial function and vascular permeability. PMID- 2980117 TI - Observations on cell kinetics and viability of a human melanoma cell line exposed to dicarboxylic acids in tissue culture. AB - Cultures of human melanoma cell line B0008 were exposed to the disodium salts of azelaic acid (C9 2Na), adipic acid (C6 2Na) and dodecanediaic acid (C12 2Na) at 10(-2) M and 5 x 10(-2) M for 24 hrs. None of the diacid salts had a significant effect on growth rate or viability of the cells, at 10(-2) M for 24 hrs nor had C6 2Na any effect at 5 x 10(-2) M. At 5 x 10(-2) M for 24 hrs, both C9 2Na, and C12 2Na had a significant effect in reducing both growth and viability. These effects were accompanied by morphological evidence of cell death, and swelling of mitochondria and accumulation of lipid droplets within cytoplasm of still viable cells. PMID- 2980118 TI - The ovaric lobule: a histoembryological unit. AB - Microscopic study of newborn female ovaries showed a constant and characteristic architectonic pattern on the surface area of the organ cortex. This pattern consisted of units that have been called "ovaric lobules" and that are made up of the various cellular types involved in the formation of the gonad (oocytes, cells derived from surface epithelium and connective-vascular elements). These structural units might be the site where synchronization or coordination for development and maturation of the successive groups of follicles occurs in post natal life. PMID- 2980120 TI - Electron microscopic study on amyloid fibril formation in human lymph nodes. AB - The purpose of this investigation was to clarify the mechanisms of amyloid fibril formation in human lymph nodes. In our present study, amyloid deposition was observed diffusely in all compartments of the lymph nodes. The deposition form showed extremely characteristic findings in its morphological features. Namely, amyloid deposits mainly consisted of clusters of round or oval nodules. Each amyloid nodule was frequently enclosed with long-stretched cytoplasmic processes of abutting reticulum cells and/or macrophages. Amyloid fibrils often formed parallel amyloid bundles radiating to outlying sections of the nodule from the center. The amyloid bundles closely adhered to the cytoplasmic membrane of not only the abutting reticulum cells, macrophages and sinus endothelium but also to the lymphocytes and plasma cells. In the central portion of the amyloid nodules, a concentric core was also observed. The most interesting finding was the intracellular formation of amyloid fibrils in all cells, such as macrophages, reticulum cells, foreign body giant cells and lymphocytes in the process of degeneration. Some fibrils localized in the limited area of the cytoplasm and others appeared in all parts of the cells, including the nucleus. Their cell membranes were missing in several areas and the cell organella had gradually dissolved. Finally the cell residuums were completely replaced by amyloid fibrils and transformed into a nodular structure with radiating bundles of amyloid fibrils. PMID- 2980119 TI - Prognostic factors in renal cell carcinoma. AB - We studied 569 cases of renal cell carcinoma in the files of the Department of Pathology of the Norwegian Radium Hospital from 1964 to 1974. A nephrectomy had been performed in all cases. Clinical information on sex, age, survival time and metastases was traced. The histological slides were examined and tumour growth pattern, cell type, cell shape, nuclear atypia, abnormal nucleoli, nuclear grade, vascular invasion and tumour demarcation were all evaluated. Besides well-known prognostic factors such as tumour stage, presence or absence of metastases and vascular invasion, nuclear grade was found to be a useful prognostic factor. Younger patients were found to do better than older, and women better than men. Smaller tumours carried a better prognosis than larger and clear cell tumours had a better prognosis than those composed of eosinophilic or basophilic cells. The presence of spindle cells was a bad prognostic omen. PMID- 2980121 TI - The neurotoxic effect of monosodium glutamate (MSG) on the retinal ganglion cells of the albino rat. AB - Monosodium glutamate (MSG) administered postnatally to the albino rat causes extensive destruction of the retina. This MSG effect does not result in complete blindness. Ganglion cells surviving the MSG treatment are healthy and functional. Using retrogradely transported HRP and Nissl staining in whole mounted retinas, it was found that the ganglion cells left after MSG treatment are not smaller than those in controls, that these cells do not belong to one cell size group, and that no cells size group is selectively missed. The results explain why photic entrainment of MSG treated animals is still possible. PMID- 2980122 TI - Non Hodgkin's lymphomas following chemoradiotherapy for Hodgkin's disease. Two new cases and a review of the literature. AB - Two patients developed non-Hodgkin's lymphoma (NHL) six and ten years after radiotherapy and chemotherapy for Hodgkin's disease nodular sclerosis type. The histological classification of the developing NHL for the two patients was: IgG (K) secreting lymphoplasmacytoid lymphoma of the stomach, and immunoblastic lymphoma of the cervical lymph nodes. Both patients responded well to conventional chemotherapy for NHL and are alive 22 and 5 months after the diagnosis of the secondary tumor. Forty eight cases of NHL after treatment for HD have been previously reported. We present a review of the literature of these cases, adding to this literature the first reported case of gastric lymphoplasmacytoid lymphoma under such circumstances. PMID- 2980123 TI - Effects of mellitic acid (MA) and sodium fluoride (NaF) on the histological appearance of murine fetal tibiae cultured in vitro. AB - The aim of this study was to develop a standardized image analysis method for localization and quantitative measurement of calcified structures of murine fetal tibiae cultured in vitro as a completion and verification of previous biochemical studies. The calcified structures of bone stained by von Kossa silver technique and the epiphyseal cartilages showing intensive metachromasia with toluidine-blue staining were converted with grey-value window programs and afterwards the areas of the selected structures were measured. The histomorphological investigations showed that the murine tibiae, incubated for a period of 6 days in a medium with addition of 5 mmol mellitic acid, showed both a significant reduction of calcium deposits and an increase of epiphyseal intercellular cartilage matrix. The tibiae incubated in a medium with addition of 0.5 mmol sodium fluoride significantly showed an increase of calcium deposits in the thickened lamellae of the compacta. These histomorphological results confirm previous biochemical studies. PMID- 2980124 TI - Growth hormone and prolactin immunoreactivity in the pituitary gland of postnatal little (lit) mice. AB - Homozygous little (lit/lit) mutant mice exhibit a growth lag which is manifested at approximately two weeks postnatally. Functional aspects of the development of pituitary growth hormone (GH) cells and prolactin (PRL) cells were thus analyzed by means of colloidal gold immunocytochemistry at the ultrastructural level in lit/lit mice and their normal counterparts ranging in age from 5 days postnatally to adulthood. In the adult normal and lit/lit pituitaries, secretory granules in GH cells and PRL cells showed a positive immunoreaction to their respective antisera, as did granules in both cell-types at 5 days postnatally. By 14 days some GH cells in lit/lit pituitaries appeared to be less densely populated with granules than GH cells in normal pituitaries, but a positive immunoreaction continued to occur even in sparsely granulated GH cells. PRL cells showed ultrastructural features in lit/lit pituitaries which were similar to those in normal mice, and immunoreactivity was present at all stages examined. The results indicate that since differences in granule reactivity were not evident between lit/lit and normal GH cells, despite ultrastructural morphologic differences which were present by 14 days postnatally, manifestations of the defect in lit/lit may be primarily quantitative in terms of numbers of granules and/or numbers of GH cells. With respect to PRL cells, neither morphologic nor functional aberrations could be observed; thus, a deficit in PRL hormone production might be the result of a more subtle defect than that in GH cells. PMID- 2980125 TI - The thoracic sympathetic neurons of the chick: normal development and the effects of nerve growth factor. AB - The generation and degeneration of sympathetic neurons in the third thoracic ganglion (segment 19) of the chick were studied between embryonic days (E) 7-18 using 3H-Thymidine autoradiography and routine cell counts. Cumulative radiolabelling experiments indicated that few sympathetic neurons were generated on E6-7. 10% of the sympathetic neurons were generated on E8 and a further 20% on E9. The final 70% of neurons completed the mitotic cycle between E10-12. Cell counts demonstrated that the neuronal population increased from 10,166 +/- 423 (mean +/- SEM) to 22,291 +/- 767 between E8-10 and remained stable up to E14. The population subsequently declined by 37%, to 14,157 +/- 831, by E18. Pyknotic neurons were found at all stages of development, but were most apparent between E7-15. The effects of Nerve Growth Factor (NGF) on the number of both surviving and pyknotic neurons in the ganglion were also examined. E9 embryos treated with NGF from E5-8 showed a 57% increase in the number of sympathetic neurons. This increase therefore occurred prior to the decline in neuronal number and was not accompanied by a decrease in the number of visibly pyknotic neurons. It is therefore possible that early NGF treatment increases the number of sympathetic neurons through a mechanism other than the attenuation of cell death. PMID- 2980126 TI - The association of medial collagenous tissue with atheroma formation in the aging human aorta as revealed by a special technique. AB - A new technique which brilliantly colors collagen fibers in a field of polarized light reveals that during mid-life the smooth muscle cells in the tunica media of the human aorta begin to disappear. The connective tissue is divided between two regions; one below the subintimal layer and the other under the adventitia. Fine collagen fibers extend upward from the former into the subintima and beyond into the intima and the overlying atheromatous plaques of the aging aorta. Thus, the source of fibrous thickening of the vessel is not confined solely to the intimal layer; at least, a portion of the total collagen content arises deep within the aortic wall. PMID- 2980127 TI - Diazo-reaction positive substance observed in the cortex of Chattonella antiqua. AB - In order to investigate the causative factors responsible for removal of mucous coat from the gill lamellae of young yellowtail, Seriola quinqueradiata by red tide, diazo-reactions were employed for planktons and their media. The concentration of NO2- in the medium containing the raphidophyceae, Chatonella antiqua (ca 2000 cells/ml), was 0.70 +/- 0.05 (mu g/ml +/- SEM). In addition, diazo-reaction positive substances (NOx) which may degenerate the mucous, was highly concentrated in the cortex (perikaryon) of Chattonella antiqua. Morphologically, mucocysts, and chloroplats were likewise present in the cortex. Mucocysts were packed with fine fibrous content. Histochemically, the mucocysts were stained with PAS and had an abundance of nitrogen oxides (NOx). We observed discharge of the fibrous material from the mucocysts. These results suggest that when Chattonella antiqua is passing between the gill lamellae, NOx discharged from the mucocysts may act on the mucous, leading to the degeneration and concomitant removal of the mucous coat from gill lamellae. PMID- 2980128 TI - Effects of Friend leukemia virus (FLV) inoculation in F1 mice and differentiation of FLV-induced leukemia. AB - Effects of Friend leukemia virus (FLV) inoculation in F1 specific pathogen free (SPF) mice were examined. Resistance to FLV was dominantly inherited both in F1 hybrid mice (BDF1) (FLV-resistant & FLV-sensitive with polycythemia) and F1 hybrid mice (B6C3F1) (FLV-resistant & FLV-sensitive with anemia). But the population dynamics of the nucleated cell components of F1 mice after FLV inoculation differed from those of FLV-resistant inbred mice. A small number of mature erythroblasts appeared in the peripheral blood of BDF1 mice. In B6C3F1 mice, erythroblastosis with splenomegaly and polycythemia occurred. However, all of these findings in BDF1 and B6C3F1 mice regressed spontaneously. In F1 mice, FLV induced an intermediate reactive pattern of the two patterns that had been induced in the parental strains. The results indicate that FLV may induce leukemia with various degrees of differentiation, according to the genetic difference of the host. PMID- 2980129 TI - Function of the follicular dendritic cell in the germinal center of lymphoid follicles. AB - The authors made an immunocytochemical examination of the germinal centers (GCs) of (1) lymph follicles in physiological lymph nodes and (2) extra-nodal tissues of divergent diseases including thyroid disorders, rheumatoid arthritis, Warthin's tumor and Kimura's disease (Eosinophilic lymphfolliculoid granuloma). In these germinal centers, the presence of immunoglobulins (IgG and IgM), early acting complement components (C1q, C4, C3c, C3d), receptors for C3b and C3d and dendritic reticulum cell-1 was demonstrated in lace-like network patterns which were proven electron-microscopically to coincide with the surface of follicular dendritic cells. IgE was distributed in a lace-like pattern in the GCs of proliferating follicles of Kimura's disease, in which the lysis of follicles was frequently observed. This lysis appeared to be related to the presence of complement components. In the germinal centers of extra-nodal tissues, including the thyroid tissues accompanying the lymph follicles, rheumatoid arthritis synovial tissues as well as Warthin's tumors, thyroglobulin, rheumatoid factor and salivary amylase were detected as specific antigens, occurring in lace-like patterns. It is possible that follicular dendritic cells may play a role in the genesis of GCs and be responsible for the immune response through C3 receptors. PMID- 2980130 TI - Duodenal microanatomy of the domestic cat (Felis catus). AB - Duodenal samples were taken from similar locations in six cats, processed, stained, and examined via light microscope. There were no prominent circular folds (plicae circulares) or stratum compactum (lamina subglandularis). The 1072 microns x 201 microns villi were covered by 46 microns high columnar epitheliocytes proximally which decreased in height (41 microns) distally and displayed a 1.1-1.7 microns striated border. Globular leukocytes, mononuclear cells, and twenty-eight goblet cells (exocrinocytus calciformis) per villus were seen. The intestinal gland (crypt of Lieberkuhn) epithelium was 20 microns tall and had a less distinct striated border. The 515 microns simple straight tubular intestinal gland layer displayed distal branching. Many mitotic figures, 12 goblet cells per gland, and occasional columnar to triangular cells with red cytoplasmic granules were seen. The thickness of the lamina propria mucosa (glandular portion) decreased from proximal to distal (563-465 microns). The lamina muscularis mucosa had two layers and decreased in thickness distally (71 28 microns). The proximal muscularis mucosa was penetrated by the ducts of submucosal (Brunner's, duodenal) glands. The tela submucosa decreased in thickness distally (593-192 microns) and contained submucosal glands with 11.5-75 microns lumina for the first 1.5-2.5 cm. However, submucosal glands could be found to a distance of 8 cm. The glandular epithelium ranged from 7.5-22.5 microns in height. Only one type of secretory cell was observed, with both mucous and serous properties. The tunica muscularis ranged from 190-1425 microns (median thickness of 557 microns) and had two layers. PMID- 2980131 TI - Metaplasia of the parietal layer of Bowman's capsule: a histopathological survey of the human kidney. AB - Human kidney sections taken at autopsy were examined to determine the incidence of metaplasia of the Bowman's parietal epithelium. Autopsy records were consulted to determine if there was any correlation between clinical disease, histopathological changes in organ systems and metaplasia of Bowman's capsule. The sections represented both sexes in 9 age groups from 2 to 87 years. The sections were fixed in neutral formalin, embedded in paraffin, sectioned at 6 microns, and stained with hematoxylin and eosin. A total of 129 kidney sections, representing 129 individuals, were evaluated. One hundred renal corpuscles were counted per section and the parietal layer of Bowman's capsule was classified as normal (squamous) or metaplastic (cuboidal). Of the 129 kidneys examined, 69 (53%) had metaplasia of Bowman's capsule. Of the 87 male kidneys, 51 (59%) exhibited metaplasia of Bowman's capsule. Of the 42 female kidneys examined, 18 (43%) of the kidneys had metaplasia of Bowman's capsule. On average, in kidneys with metaplasia, 4% of the renal corpuscles had metaplasia of Bowman's parietal layer. The lesion was present in both sexes in all age groups. The autopsy records revealed that there was no common clinical condition associated with the metaplastic lesion, but metaplasia of Bowman's parietal epithelium was consistently present with hepatic congestion and/or fatty changes. PMID- 2980132 TI - Ultrastructural changes in pancreatic acinar cells of rats after administration of 4-hydroxyaminoquinoline-1-oxide. AB - Ultrastructural changes in the exocrine pancreas 24, 48, 72 and 168 hours after a single intravenous injection of 4-hydroxyaminoquinoline-1-oxide (4-HAQO), at a dose of 14 mg.per kg., were observed in male rats. At 24 hours after administration, multiple focal degenerative lesion-like large vacuoles in the acini, and a decrease in zymogen granules with dilation of the rough endoplasmic reticulum cisternae in the acinar cells were marked. At 48 hours, acinar cell degeneration and necrosis progressively increased. The nucleus, especially, appeared to be disorganized and lysosome-like bodies with various sizes were frequently observed. At 72 hours, acinar cell degeneration persisted in the acini, and the interstitial space with infiltration of inflammatory cells appeared edematous. In addition, ductular-like cells, which resembled intercalated duct cells, possessing a light cytoplasm with occasional mitoses were observed around the duct lumen. At 168 hours, the exocrine pancreas was occupied with proliferated ductular-like cells. Furthermore, acinar cells and acini regenerated to the normal pattern were sometimes found. Thus, the exocrine pancreas degenerated progressively up to 48 hours after administration of 4-HAQO, gradually came to be repaired by degrees from 72 hours and then partly appeared to be regenerated at 168 hours. It is suggested that the ductular-like cell might be the precursor of the acinar cell in the regenerating process after injection of 4-HAQO. PMID- 2980133 TI - Occurrence of pancreatic ductal cell dysplasia in rats fed with a high fat diet and ethanol. AB - The effects of alcohol and diet on acute pancreatitis were studied in 192 male Wistar rats. The animals were fed with standard laboratory food up to three months of age and, after that, were divided into four groups of 48 animals, each group receiving a different diet: standard, fat-rich, protein-rich or carbohydrate-rich. In each diet group, 24 animals obtained 15% (v/v) ethanol in their drinking solution while the other 24 rats had water ad libitum. The diet period lasted for 12 weeks, after which acute experimental pancreatitis was induced under diethyl ether anesthesia by ductal injection of rat bile into the pancreatic ducts. Moderate or severe ductal cell dysplasia developed in three of the 15 survivors in the group fed with a high-fat diet and 15% ethanol in their drinking solution. Mild acute pancreatitis was histologically found in 13 rats and moderate pancreatitis in one rat in this group. One rat did not show any pancreatic parenchymal changes. Two of the rats with ductal cell dysplasia had mild pancreatitis and the pancreas of the third rat was normal in this respect. Dysplastic changes were not found in any other experimental group used in the study. The observation is statistically significant at p less than 0.025 level. The results indicate that alcohol and a high fat diet together might have a carcinogenic effect on pancreatic ductal epithelium in rats. PMID- 2980134 TI - Dying and regeneration of human tumor cells after heterotransplantation to athymic mice. AB - The histologic phenomena occurring immediately after heterotransplantation of two human colon adenocarcinomas to athymic mice have been studied. The tumors differed with respect to velocity of growth and passage age. Three phases were discernible in both cases. (1) During the first phase, most inoculated tumor cells died. (2) The second phase was characterized by removal of the necrotic tumor cells by immigrated inflammatory cells and by penetration of the connective tissue of host animals from peripheral into central areas of the implants. The first mitoses occurred within tumor cells in close proximity to these connective tissue septa. (3) During the third phase, signs of regeneration and proliferation of tumor cells resulted in the macroscopic enlargement of xenografts. Only in this phase, the typical histologic characteristics of the tumors were formed. These observations point to the host connective tissue invading into implants to be of great importance for the stimulation of tumor cell proliferation and, therefore, for the growth of xenografts. Thus, successful heterotransplantation is obviously based on mutual events between the transplanted tumor cells and host connective tissue. PMID- 2980136 TI - Bulging of lumbar intervertebral discs: non-contacting measurements of anatomical specimens. AB - A noncontacting photogrammetric method was used to measure intervertebral disc bulge (deformation perpendicular to the disc surface) at an anterior site and a posterolateral site of 17 human lumbar discs. Specimens were tested in constructs consisting of two lumbar vertebrae and the intervening disc. The posterior elements were removed to give unobstructed visibility of the posterolateral aspects. Stereophotogrammetry was used to record the motion of target points attached to the disc and vertebral body surfaces. In axial loading, the bulging increased linearly with disc compression, which in turn had a nonlinear relationship with the applied load. Axial loads and offset compression loads of physiologic magnitude caused disc bulge that was generally less than 1 mm. Axial torsion of an otherwise unloaded specimen produced minimal change in disc bulge. The disc bulging measured in this study is generally less than that previously reported, especially for angular motions between the vertebrae. This is probably because the photogrammetric method used here is independent of motion of the vertebrae, which can corrupt mechanical, contacting methods of measurement. PMID- 2980135 TI - Blastic OKT6-positive proliferation preceding malignant histiocytosis. AB - A 45-year old male presented latero-cervical lymphoadenopathy. Biopsy revealed a malignant proliferation of immature "lymphoid" cells bearing T6 antigen and HLA DR but negative for other lymphoid markers, suggesting a phenotype similar to Langerhans cells. The patient did not receive any therapy and six months later developed a histologically typical malignant histiocytosis, involving spleen and liver. Other reported cases of lymphoid malignancies (two bearing the T6 antigen on blast cells) preceding malignant histiocytosis were found and compared with ours. Most of these cases were characterized by the pediatric age of the patients and were presented as acute leukemias exhibiting, in at least some of them, reliable T-cell markers. Our case appears to represent, on the other hand, a blastic proliferation of precursors of both histiocytes and Langerhans dendritic cells which eventually progressed to malignant histiocytosis. The relevance of this observation in the debate on the origin of Langerhans cells and the relationships existing between macrophages and dendritic cells is discussed. PMID- 2980137 TI - Spina bifida occulta associated with a low-placed conus medullaris. AB - From 1977 to the present, we have managed 30 patients with spina bifida occulta associated with a low-placed conus medullaris. Twenty-six patients had surgery performed. Nineteen patients demonstrated urological and neurological improvement. The characteristics of this condition include a cleft spine revealed by plain x-ray, vesicorectal disturbances, local findings (such as a tumor, dimpling, and angioma in the lumbosacral region), and deformities of the feet. Surgery is indicated after confirmation of neurological symptoms, such as a vesicorectal disturbance, after about 1 year of age or older. Operation is not needed immediately after birth. PMID- 2980138 TI - Experimental approach--and literature review--of spinal care in adults with a neuromuscular disorder. PMID- 2980139 TI - Spinous process segmental instrumentation for scoliosis. AB - Spinous process segmental instrumentation (SPSI) for spinal fusion was devised in 1983 by Drummond et al. in an attempt to achieve the stability of segmental fixation without the known neurologic risks of passing sublaminar wires. We used SPSI in 75 scoliosis patients. Sixty-one had idiopathic scoliosis, 12 had neurogenic scoliosis, and 2 had congenital scoliosis. There were no deep infections, pseudarthroses, or neurologic complications. Two patients experienced upper hook dislodgement with 10 degrees loss of correction. We concluded that SPSI can achieve the correction of Harrington rod instrumentation and the stability of Luque rod segmental instrumentation, without the neurologic risk of sublaminar wiring. PMID- 2980140 TI - Evaluation of spinous process wire fixation with Harrington instrumentation for idiopathic scoliosis. AB - A modification of the technique of Harrington instrumentation for idiopathic scoliosis utilizes segmental spinous process wiring. The purpose of this study is to contrast and compare two populations of surgically treated patients with idiopathic scoliosis: one group with a single Harrington distraction rod and the other group with the addition of spinous process wiring. Of 252 consecutive patients between 1971 and 1987, 215 were retrievable, with an average clinical follow-up of 2.2 years. Curves were analyzed by location and patient age. Patients treated with spinous process wires were braced, those without were casted. No significant difference in terms of percent correction with time was evident between the two treatment groups. The complication rate (11.0%) and pseudarthrosis rate (4.0%) were the same in both wired and standard groups. The more rigid adult curves resulted in less correction and greater complications than adolescent curves. Spinous process wiring seemed to protect against upper hook cut-out. There were no deaths or paraplegias. Patients surgically treated with the wire modification enjoyed greater comfort with the removable brace, and curve correction was equal to that obtained in patients treated with the standard technique and post-op casting. PMID- 2980141 TI - Lower extremity fractures in paraplegics: pattern, treatment, and functional results. AB - Eighty-seven lower extremity fractures in 71 paraplegics were retrospectively evaluated to determine adequacy of treatment, functional result, and range of motion of the hip and knee. Most fractures treated closed healed without difficulty, but with a complication rate of 19%. Length of hospitalization averaged almost 3 months in the group treated closed, and residual hip and knee stiffness was the major complication. This was not a problem except in high demand patients, such as wheelchair athletes, in whom decreased range of motion was a significant limitation. Treatment with circular casts produced a high number of complications and is not recommended for patients with insensate skin. Open reduction and internal fixation is recommended for a small subgroup of paraplegics: wheelchair athletes, hip fractures, and those in whom autonomic dysreflexia and/or uncontrollable spasticity due to fracture manipulation is a significant hazard. PMID- 2980142 TI - Pyogenic osteomyelitis of the spine--a review of 16 consecutive cases. AB - Pyogenic osteomyelitis of the spine is an uncommon disease. A series of 16 cases is reported. The site of involvement includes the lumbar, cervical, and thoracic segments, in that order of frequency of occurrence. Drug addiction and diabetes mellitus are important predisposing factors. The spectrum of clinical presentation is described. Late presentation is usually masked by old age, debilitation, and previous antibiotic treatment. Rapid progression with septicemia is also encountered. The importance of differentiating this condition from tuberculosis spondylitis is emphasized. Definitive diagnosis should depend on bacteriological, histological, and, with the recent introduction of antituberculosis antibody test, serological studies. Timely surgical intervention is indicated in complicated cases. Radical excision and anterior spinal fusion yields early and complete eradication of the infected material plus arthrodesis, which prevents late collapse of the spine. PMID- 2980143 TI - Diagnostic lumbar nerve root block. AB - Lumbar nerve root anesthesia using a local anesthetic was performed in 100 cases of sciatica. The neurological state before and after nerve block was recorded and also the degree of pain alleviation caused by the block. Three indications for the block were used in the study: unilateral sciatic pain and normal findings on myelography and/or CT or MR (n = 51), minor myelographic findings that possibly but not necessarily explained the patient's symptom (n = 40), and multiple pathological findings on myelography (n = 9). In 87% of the patients, the nerve block inflicted temporary sensory and/or motor disturbance within the dermatome anesthesized. In total, 51% experienced total, 31 partial, and 18 no pain relief after injection. No side effects of the procedure were noted. Patients with pain alleviation after anesthesia were offered an operative nerve root decompression and short-term results of this decompression seem to be comparable to conventional disc surgery. Anesthetizing the lumbar nerve root outside the intervertebral foramen may be considered in the preoperative evaluation of patients with sciatic pain and minor or no radiographic findings. PMID- 2980144 TI - Disc space infection. PMID- 2980145 TI - Formula of specificity in evoked potential monitoring. PMID- 2980146 TI - Occupational health sciences in the 1990's. PMID- 2980147 TI - Molecular mechanisms of chemical toxicity. PMID- 2980148 TI - Genetic screening--an absolute necessity or a misconception? PMID- 2980150 TI - Programme of occupational health research in Poland (1986-1990). PMID- 2980149 TI - Toxic effects of combined exposure to toluene and xylene in animals. I. Acute inhalation study. PMID- 2980151 TI - Teratogenicity, fetal and placental toxicity of 1,3,5-trioxane administered to pregnant female rats. PMID- 2980152 TI - The effects of physical effort on pulmonary uptake of selected organic compound vapours. PMID- 2980153 TI - Evaluation of SCEs in bone marrow cells of mice after in vivo exposure to asbestos. PMID- 2980154 TI - A mortality study among male workers occupationally exposed to asbestos dust in Poland. PMID- 2980155 TI - Radiation exposure of miners and systems of radiation protection in Polish nonuranium mines. PMID- 2980156 TI - The role of some cyclorrhaphan flies as carriers of human helminths in Malaysia. AB - The role of some adult flies (Diptera: Cyclorrhapha) as carriers of helminth parasites of man was studied at four sites in Malaysia: a refuse dump, where no helminth-positive flies were detected, and in three peri-domestic situations where four species of flies carried up to three types of nematodes. The dominant fly species Chrysomya megacephala (Fabricius) carried eggs of the roundworm Ascaris lumbricoides L., the pinworm Trichuris trichiura (L.) and hookworm on the adult external body surface and in the gut lumen, in association with Bukit Lanjan aborigines. Chrysomya rufifacies (Macquart) and Sarcophaga spp. also had Ascaris lumbricoides and Trichurus trichiura eggs in their gut contents. Human helminths were not recovered from Lispe leucospila (Wiedemann), Lucilia cuprina (Wiedemann) or the housefly Musca domestica L. In an urban slum area of Kuala Lumpur city, filariform larvae identified as the hookworm Necator americanus (Stiles) occurred in the intestines of the face-fly Musca sorbens Wiedemann (22 larvae per 100 flies) and of Chrysomya megacephala (4.5 larvae per 100 flies). This concentration of apparently infective N. americanus in M. sorbens, a fly which often breeds in faeces and browses on human skin, could have transmission potential. PMID- 2980157 TI - Juvenile hormone mimics as effective sterilants for the tsetse fly Glossina morsitans morsitans. AB - The development of puparia of Glossina morsitans morsitans Westwood was disrupted by topical applications of the juvenile hormone mimics S-methoprene (the resolved enantiomer of 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoic acid 1-methyl ester) (Zoecon), S21149 (propionaldoxime-0-4-phenoxyphenoxyethylether) (Sumitomo), or S31183 (2-[1-methyl-2-(4-phenoxyphenoxy)ethoxy]pyridine) (Sumitomo) dissolved in acetone. Puparia so treated during the first 4 days of life suffered developmental abnormalities, the severity of which were dose-dependent. Similarly, puparia produced by adult females treated with these compounds were abnormal. Dose-response data showed that effects were greatest with S31183 and least with S-methoprene. Abnormalities in the form of abdominal lesions and wing crumpling were typical of flies emerging from puparia produced by S-methoprene treated females. However, arrested development at the red eye and pigmented seta stage within the puparium were typical of offspring of females treated with S21149 and S31183. A dose of 2 micrograms per female of S31183 was sufficient to prevent emergence of offspring produced for the rest of the life of the fly. The same dose resulted in partial recovery of females treated with S21149 some 18 days following treatment. Treatment with 2 micrograms S-methoprene did not suppress completely the production of normal offspring and recovery was complete some 27-35 days after treatment. Exposure of males to 20 micrograms S31183 did not impair their ability to inseminate females; transfer of material during copulation was sufficient to prevent the production of viable offspring by their mates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980158 TI - Reduced susceptibility to permethrin and its relationship to DDT resistance in larvae of Anopheles stephensi. AB - Permethrin selection of DDT resistant Anopheles stephensi Liston mosquito larvae produced a reduction in susceptibility to knockdown (2 h exposure) of 17-fold, but only 1.6-fold to kill (24 h exposure). Genetic analysis, incorporating visible mutant markers, was interpreted as indicating that, through multigenic inheritance, several interacting genetic factors were collectively responsible for reduced larval susceptibility to knockdown. These were maintained together only under selection pressure, as the effect was lost quickly in the absence of selection or with outcrossing. The 30-40-fold DDT-resistance found in the parental strain was barely altered by permethrin selection, suggesting no relationship with the major source of DDT resistance. This was confirmed in single family studies. Some evidence for an additional tolerance to DDT was found to be associated with reduced larval susceptibility to permethrin. PMID- 2980159 TI - The phlebotomine sandflies of Venezuelan Amazonia. AB - Phlebotomine sandflies were surveyed in two ecologically contrasted areas, the northern moist and southern wet tropical forests, of the Territorio Federal Amazonas, Venezuela. Three new taxa and twenty-one new records were added to the previously known species list for Venezuelan sandflies, which now totals eighty species. Both sexes of Lutzomyia (Psychodopygus) killicki sp.n., L. (Trichophoryomyia) bettinii sp.n., L. (Nyssomyia) olmeca reducta subsp.n. and and the females of L. bernalei Osorno et al., Brumptomyia pintoi Costa Lima and L. begonae (Ortiz & Torres) are described and illustrated. PMID- 2980160 TI - Leishmaniasis in Sardinia. III. Soil analysis of a breeding site of three species of sandflies. AB - Soil analysis was carried out on samples obtained from a breeding-site of the sandflies Phlebotomus perniciosus Newstead, P. perfiliewi perfiliewi Parrot and Sergentomyia minuta (Rondani) in Sardinia. Soil texture, pH, CaCo3, organic matter and water content showed no correlation with the number of sandflies that emerged from the places where soil samples were taken. It appears that pre imaginal stages of these phlebotomine sandflies are associated with a relatively stable, cool and humid environment protected from sunshine and rain, rich in clay and in organic nitrogen. PMID- 2980161 TI - In vitro haemagglutination and attenuation of microfilarial motility by haemolymph from individual blackflies (Simulium ornatum) infected with Onchocerca lienalis. AB - Two in vitro tests were used to investigate the effect of Onchocerca lienalis Stiles infection on the haemolymph of Simulium ornatum Meigen. The first of these examined the effect of infected haemolymph on the motility of fresh O. lienalis or Brugia pahangi Buckley & Edeson microfilariae. Incubation of haemolymph from individual flies with fresh microfilariae was performed in the wells of Terasaki micro-tissue culture plates. Motility of both species of parasite was found to be significantly attenuated when compared to worms incubated in control haemolymph groups. The second assay was that of agglutination of cat erythrocytes in the presence of haemolymph from individual flies, also performed in Terasaki plates. This test demonstrated significant increases in the rates of haemagglutination in the haemolymph of O. lienalis infected blackflies. The titre appeared to increase during the initial 5 days of infection up to a level of 1/32+, but then fell between day 5 and 7 to a maximum level of 1/2. The proportion of flies exhibiting haemagglutination also rose following infection. Despite the apparent absence of melanization and encapsulation, simuliids may have at least two humoral haemolymph components available to them for parasite regulation; a fast-acting factor responsible for rapid parasite death, and more specific agglutinins, possibly lectins. The role of the latter in defence is as yet unclear. PMID- 2980162 TI - Selective attractiveness of rodent-baited traps for female blowflies. AB - Five species of Calliphoridae, mostly females, were collected using fly-traps baited with freshly killed rodent carcasses. Female blowflies were examined to determine the stages of ovarian development. The traps were selective for certain ovarian stages which varied according to the species. Phaenicia eximia (Wiedemann) was attracted to the traps mainly to oviposit, whereas females of Chrysomya albiceps (Wiedemann), C. putoria (Wiedemann) and C. megacephala (Fabricius) were captured in intermediate stages of oogenesis but seldom with eggs mature for oviposition. It is concluded that this kind of baited trap is unsuitable for the collection of male blowflies and that samples of female blowflies are specifically biased in favour of certain ovarian conditions. PMID- 2980163 TI - Use of a male-specific DNA probe to distinguish female mosquitoes of the Anopheles gambiae species complex. AB - A method has been developed to distinguish between mated female individuals of the sibling species Anopheles gambiae Giles sensu stricto and Anopheles arabiensis Patton. The DNA probe pAna1, reported by Gale & Crampton (1987a) to be useful for the specific identification of An. arabiensis males, is here shown to be sufficiently sensitive to deduce the species identity of inseminated females from the identity of the sperm contained within the spermatheca. PMID- 2980164 TI - Anti-mosquito antibodies decrease the reproductive capacity of Aedes aegypti. AB - Aedes aegypti (L.) fed on rabbits immunized with mosquito antigens showed a reduction in fecundity in the first oviposition and decreased viability of the progeny. Feeding behaviour of mosquitoes was not affected and no significant mortality was observed due to the presence of anti-mosquito antibodies in the bloodmeal. Antibodies were detected in the oocytes of mosquitoes 48 h after the bloodmeal. The role of specific antibodies in influencing fecundity is discussed. PMID- 2980165 TI - The morphology of the egg of the bird-parasite Protocalliphora azurea (Diptera: Calliphoridae). PMID- 2980166 TI - Evidence for a 'memorized' home range in Anopheles farauti females from Papua New Guinea. AB - Evidence is presented that female Anopheles farauti Laveran released in an alien village dispersed more, in their first oviposition cycle after release, than females released in their village of capture. In a subsidiary experiment transporting the mosquitoes did not affect dispersal; wind speed or direction was not sufficient to account for the phenomenon. It is hypothesized that these permanent-pool breeding mosquitoes make appetitive long-range flights to oviposition sites. Mosquitoes blood-fed before midnight had a shorter oviposition cycle than those fed just before dawn. Nulliparous females, and those with well defined ovariolar dilatations, predominated in the early evening whereas females that had oviposited recently were largely collected in the middle and later parts of the night. PMID- 2980167 TI - Allergens of non-biting midges (Diptera: Chironomidae): a systematic survey of chironomid haemoglobins. AB - Various genera of non-biting midges (Diptera: Chironomidae) possess haemoglobins in larvae and adults. For certain species, these haemoglobins have been implicated in human allergic disease in several countries. The present study confirms and extends observations that haemoglobin is present in many species of Chironomidae, establishes that it is retained from the larval to the adult stage and shows that haemoglobin persists both in live and in dead dry flies. Previous suggestions that Chironomidae should be seen as important environmental and occupational allergens are clearly substantiated. PMID- 2980168 TI - Equine Culicoides hypersensitivity in Florida: biting midges collected in light traps near horses. AB - Twenty-three species of Culicoides were trapped near pruritic horses during a 2 year survey in Florida. Nearly 99% of the biting midges collected were represented by Culicoides insignis Lutz, C. edeni Wirth and Blandon, C. stellifer (Coquillett), C. niger Root and Hoffman, C. haematopotus Malloch and C. venustus Hoffman. The relative contribution to the total catch by each of these species varied among collection sites. Seasonally, different species attain their largest population sizes at different times. Association of species collected in light traps with the seasonality of lesion development on the horses suggests that more than one species must be involved in causing Culicoides hypersensitivity in horses from Florida. Nulliparous females were present in most months that females of the same species were active; in some cases, species were trapped in each month of the year, particularly at the more southerly sites. The species of biting midge most likely to be involved in Culicoides hypersensitivity in horses in Florida are C. insignis, C. stellifer and C. venustus based upon their mammalophilic behaviour and seasonality. PMID- 2980169 TI - Early season dispersal of Muscidifurax zaraptor (Hymenoptera: Pteromalidae) utilizing freeze-killed housefly pupae as hosts. AB - The pteromalid wasp, Muscidifurax zaraptor Kogan and Legner, was released at three locations at a dairy in May before housefly and stable fly breeding had begun. Freeze-killed housefly pupae were placed adjacent to the emerging parasites at biweekly intervals for a 6-week period. Hosts placed out weeks 0 and 2 were heavily parasitized. Decreased parasitism in hosts placed out at week 4 suggested that many of the M. zaraptor had dispersed or died. High parasitism of hosts placed in the field at week 6 was the result of second generation parasites emerging from pupae placed out at week 0. Parasitism of freeze-killed housefly pupae placed 6 m and in the four cardinal directions from the release points was similar but lower than for hosts placed adjacent to the emerging parasites. The study demonstrated that emerging M. zaraptor readily utilized nearby freeze killed housefly pupae but the availability of these hosts did not deter the parasites from searching for additional hosts. PMID- 2980170 TI - Morphology and identification of Chrysops larvae from Nigeria. AB - Mature larvae of four Afrotropical species of Chrysops (Diptera: Tabanidae): C. centurionis Austen, C. distinctipennis Austen, C. longicornis Macquart and C. silaceus Austen, are described and illustrated from specimens collected in Nigeria, identified by rearing associated adult flies. The descriptions pay special attention to the value of ultrastructural characters, such as cuticular striations, observed by scanning electron microscopy (SEM). PMID- 2980171 TI - Physiological characteristics of Culex pipiens populations in the Middle East. AB - We determined whether exposure to a short day-light regime of 14 h 15 min induces ovarian diapause in Culex pipiens L. mosquitoes from Israel and whether differences occur in certain morphological, physiological and behavioural traits. Samples from nineteen localities in Israel, from 33 degrees 05' N to 29 degrees 35' N latitude, conformed to the morphological criteria of Cx pipiens, sensu stricto (i.e. wider spread of dorsal than of ventral arms of the aedeagus) as determined by negative DV/D ratios of the male. Autogenous females occurred at frequencies of 4-55% in all areas of Israel throughout the breeding season. No mating barriers were detected between individuals of autogenous and anautogenous genotypes. Autogenous females were no more prevalent from polluted or enclosed breeding sites than from others with various degrees of openness. In general, Cx pipiens females fed equally well on human (35% engorgement) and avian (22% engorgement) hosts (P less than 0.01). Although females from the southern part of the study region appeared to be incapable of ovarian diapause, at least some of those from the north experienced diel-mediated diapause. We conclude that, in the Middle East, the 33rd parallel provides a southern limit to the range of Cx pipiens with capability for diel-mediated ovarian diapause, but that non diapausing Cx pipiens s.l. are present at least as far south as Elat (20 degrees 25' N) on the coast of the Gulf of Aquaba. PMID- 2980172 TI - Use of fluorescent pigments for the automatic marking of tsetse flies in traps. AB - A means of contaminating tsetse flies in the field with fluorescent pigment powders has been developed, using pigment in open-ended plastic chambers at the cage position on traps. Glossina pallidipes Austen and G.morsitans morsitans Westwood passed rapidly through the chambers, and on exit were contaminated with consistent doses of powder: about 90 micrograms/fly when powder was presented on the chamber roof and about 28 micrograms/fly when powder was presented on the chamber floor. The technique automatically marks tsetse flies with pigment, cheaply, simply and with the minimum imposition of stress and is expected to be particularly useful in ecological studies. Its potential for marking other biting flies is discussed. PMID- 2980173 TI - Landing periodicity of the mosquito Aedes aegypti in Trinidad in relation to the timing of insecticidal space-spraying. AB - Diel landing periodicity (biting cycle) of domestic Aedes aegypti (L.) in Trinidad, West Indies, was monitored using human bait during January-August 1980. The periodicity of females was predominantly diurnal (95.2% arriving during daylight or twilight) and bimodal, with consistent peaks at 06.00-07.00 and 17.00 18.00 hours. The diel periodicities at indoor and outdoor sites were virtually identical. Larger numbers of adults were collected outside than inside houses. It is recommended that the time of insecticidal ULV adulticiding should coincide with peaks in landing periodicity of the Ae.aegypti adults. PMID- 2980174 TI - Effects of 'closed' houses on the Aedes aegypti eradication programme in Trinidad. AB - The effects of closed (i.e. locked) houses on the efficiency of control measures against Aedes aegypti (L.) were monitored weekly for 1 year (March 1981 to February 1982) using ovitraps in the towns of Felicity and Trincity, Trinidad. Regular anti-aegypti activities comprised source reduction, focal treatment with larvicidal 1% temephos sand granules and perifocal treatment with 40% wp fenthion. In Felicity, only 6% of houses were closed and after each treatment cycle ovitrap indices showed 100% control. In Trincity, 32% of houses were closed and after each treatment cycle reductions in ovitrap indices were less than 50%. Ae.aegypti eggs sampled in ovitraps totalled 493 (1-70 per positive paddle) in Felicity and 2310 (1-75 per positive paddle) in Trincity; twenty-three ovitraps were found to be positive in Felicity and 247 in Trincity. Ae.aegypti was successfully eliminated from Felicity during weeks 40-52 using focal/perifocal treatment monitored by ovitraps. Control failure in Trincity was attributed to the survival of untreated Ae.aegypti populations in closed houses. Recommendations to improve gaining access for inspection and treatment of closed houses include having control/surveillance workers commence activities at 06.00 hours on weekdays, to work at weekends or during school holidays, or as a last resort to invoke court orders against householders harbouring Ae.aegypti. PMID- 2980175 TI - Trials of compounds to enhance trap catches of Glossina palpalis palpalis in Liberia. PMID- 2980176 TI - Large-scale rearing of Glossina brevipalpis in the laboratory. PMID- 2980177 TI - Tsetse: the limits to population growth. AB - Growth rates of tsetse populations were estimated by calculating the dominant eigenvalues of appropriate Leslie matrices. The individual effects of four variables (pre-adult and adult survival probability, interlarval period and pupal duration), have been investigated by varying each one over a wide range of values, while the other three are held constant. R, the log of the growth rate, was found to vary approximately linearly with adult and pre-adult death rate; a 1% change in the adult death rate causes approximately a 10-fold change in R. R varies linearly with the log of fecundity and of the pupal duration. An increase in the pupal duration results in a decrease in the growth rate for populations which have a positive growth rate, but an increase for populations which have a negative growth rate. For a population at equilibrium, a change in the pupal duration has no effect. Small changes in fecundity have less effect on the growth rate than small changes in the death rate; this fact is advanced as an important contributor to the generally very cautious nature of female tsetse, and their aversion to man, particularly as a potential host. A simple linear model is described which relates R to all four variables and their first order interactions. The model is used to produce a set of graphs which encapsulate the relationship between the growth rate and the vital parameters over a wide range of values. It is also used to draw the loci on one side of which tsetse populations grow, and on the other of which they decline. Population resilience is discussed in relation to the problem of tsetse eradication; it is concluded that if one can impose and sustain an added mortality of 4% per day on any female tsetse population then it must go extinct, regardless of the strength of the density dependent processes; and it seems likely that in most field conditions only an added 2-3% is required. It is pointed out that ground and aerial spraying techniques produce much higher daily mortalities than this, but they may often not be sustained for sufficiently long to achieve eradication. When odour-baited targets are used the increased death rate is much smaller, but it can be sustained as required; recent work in Zimbabwe shows that there is a good correspondence between the calculated imposed death rate and the observed rate of decline of tsetse populations. PMID- 2980178 TI - Maggot therapy: a review of the therapeutic applications of fly larvae in human medicine, especially for treating osteomyelitis. AB - In traditional medical practice, the larvae of some Diptera: Calliphoridae, notably Lucilia illustris (Meigen), L.sericata (Meigen) and Phormia regina (Meigen), have been employed for maggot therapy, i.e. to help clean lesions antiseptically, especially for treatment of chronic osteomyelitis. This mode of treatment remains appropriate for cases where antibiotics are ineffective and surgery impracticable. PMID- 2980179 TI - Panveld oviposition sites of floodwater Aedes mosquitoes and attempts to detect transovarial transmission of Rift Valley fever virus in South Africa. AB - Floodwater aedine mosquito eggs were recovered from soil samples taken from grassland depressions, called pans, in the Orange Free State Province of South Africa. A sedge, Mariscus congestus (Vahl) C.B.Cl., was a useful indicator of Aedes (Ochlerotatus) juppi McIntosh oviposition areas. No transovarial transmission of virus was demonstrated by Ae.juppi females reared from the eggs and allowed to feed shortly after eclosion on hamsters. No virus was recovered from 557 pools of 5425 adult Ae.juppi that were collected as eggs and reared to the adult stage in the laboratory. Rift Valley fever virus replicated to high titres in experimentally infected Ae.juppi females, but horizontal transmission experiments proved inconclusive. PMID- 2980180 TI - The distribution of phlebotomine sandflies in the principal ecological zones of Jordan. AB - A survey of the sandflies (Diptera: Psychodidae) of Jordan increased the number of species known from the country from three to thirteen. Composition of the phlebotomine fauna and the relative abundance of sandfly species differ among the six climatic zones, with a general distinction between the dry desert zones and the more humid Mediterranean zones. Species of Phlebotomus predominate in the former and Sergentomyia in the latter. Within the desert zone some species are restricted to the eastern desert and others to the Jordan Valley in the west. PMID- 2980181 TI - Comparative study on the infection rates of different laboratory strains of Glossina species by Trypanosoma congolense. AB - Teneral Glossina morsitans centralis Machado, G.austeni Newstead, G.palpalis palpalis Robineau-Desvoidy, G.p.gambiensis Vanderplank, G.fuscipes fuscipes Newstead, G.tachinoides Westwood and G.brevipalpis Newstead, from laboratory-bred colonies, were fed at the same time on the flanks of ten goats infected with Trypanosoma congolense Broden isolated in Tanzania or in Nigeria. The seven tsetse species were infected over the range 0.3-49.2%. Survival of both T.congolense isolates was best in G.m.centralis, poorest in G.austeni and the four palpalis group tsetse, with G.brevipalpis intermediate. It is suggested that there are differences in the gut of different laboratory-bred cultures of Glossina Westwood species and subspecies such that T.congolense parasites can survive better in the gut of some than in others and undergo cyclical development to metacyclics in the hypopharynx. PMID- 2980182 TI - ELISA absorbance cut-off method affects malaria sporozoite rate determination in wild Afrotropical Anopheles. AB - Malaria sporozoite infection rates in a mixed species group of 244 Anopheles gambiae Giles sensu lato and 115 An.funestus Giles wild female mosquitoes were compared using three methods to determine cut-off absorbance values for positivity of a Plasmodium falciparum Welch enzyme-linked immunosorbent assay (ELISA). Positive controls were based on P.falciparum circumsporozoite protein. As negative controls, four wild male Anopheles were included on each microtitre plate; tests were repeated on four consecutive days for each plate. Infection rates were estimated at 13.1-22.8% using the mean absorbance value of negative controls plus three standard deviations, 11.7-12.8% using double the mean and 12.5-13.6% using the fixed cut-off value of 0.20 (allowing for 20% variation in negative control absorbance values). Observed agreement for positivity or negativity among samples tested four times was 98.6% for the 2 x mean method, 97.2% for the fixed cut-off 0.20 value, but only 82.7% for the mean + 3 SD method. It was concluded that the 2x mean cut-off method is most reliable for field studies. P.falciparum sporozoite rates of 12.2% in An.funestus and 11.9% in An.gambiae s.l. were thus determined on the basis of the 2x mean cut-off method. This comparative evaluation demonstrates that vector infectivity rates can be seriously over-estimated from sporozoite ELISA tests, by as much as 87% in one case considered here, depending on the absorbance cut-off method applied for negative controls. PMID- 2980183 TI - Feeding strategies of the horseflies Hybomitra expollicata and Tabanus bromius in southern France. AB - Blood-feeding success of female horseflies, Hybomitra expollicata Pandelle and Tabanus bromius L. (Diptera: Tabanidae), was studied. Bloodmeal weights of flies permitted different periods of residence on a horse's belly were estimated, and related to subsequent ovarian development and the number of mature eggs produced. Fitness gain per visit to a host (as estimated by the number of mature eggs produced) increased more rapidly with residence time on the host for H.expollicata than for T.bromius. H.expollicata inhibed blood more rapidly and, above a threshold bloodmeal size, produced a consistently higher number of eggs per unit weight of blood ingested. The feeding strategies of the two species are compared. PMID- 2980184 TI - The tropical African latrine blowfly, Chrysomya putoria (Wiedemann). AB - The status of the tropical African latrine blowfly Chrysomya putoria (Wiedemann, 1830) is recognized as distinct from the sub-tropical African blowfly C.chloropyga (Wiedemann, 1818) (Diptera: Calliphoridae). The biology of C.putoria in laboratory culture is described briefly with emphasis on the differences in development found in three strains of this species from Tanzania, Liberia and Brazil. All three strains were reproductively compatible and there was no evidence of F1 hybrid sterility or hybrid breakdown in subsequent generations. Larval development was significantly faster in the Brazilian strain than in the strain from Tanzania. Sexual maturity was significantly faster in adults of the Liberian and Brazilian strains compared to that found in the strain from Tanzania. This latter strain was affected by the source of protein available as adult food. These differences between strains did not appear to be due to selection in laboratory culture, but rather evidence of the evolution of physiological divergence within a species occupying different geographical regions. PMID- 2980185 TI - A field study of mating between two species of bedbug in northern KwaZulu, South Africa. AB - The interspecific cross-mating of female Cimex lectularius Linnaeus with male Cimex hemipterus (Fabricius) results in a visible mark developing in the female ectospermalege. This mark has been used to record the frequency of mating between female C.lectularius and male C.hemipterus in sympatric wild populations in a DDT sprayed area of northern KwaZulu, South Africa. A high percentage (11/16 = 69%) of C.lectularius females were marked, showing that these two species mate in nature. PMID- 2980186 TI - Production of a hybrid between the bedbugs Cimex hemipterus and Cimex lectularius. AB - Of 479 eggs laid by female Cimex lectularius Linnaeus and C.hemipterus (Fabricius) mated by males of the other species, one was fertile and gave rise to a first stage nymph. The egg showed sculpturing typical of C.hemipterus, the female parent, and the nymph conformed to the narrow pronotum and abdomen of this species, being significantly different from C.lectularius in the width of the abdomen. Because the orientation and lengths of the bristles on the sides of the abdomen were distinctly different from C.hemipterus and closely resembled C.lectularius, the single nymph obtained from the cross C.hemipterus x lectularius was interpreted as being a hybrid rather than a product of parthenogenesis. PMID- 2980187 TI - Morphogenetic diapause in Amblyomma variegatum (Acari: Ixodidae). AB - In southern Africa, Amblyomma variegatum Fabricius is characterized by a strict seasonal activity. Experiments were carried out to determine whether a diapause mechanism regulates this seasonality. Engorged A.variegatum females were exposed to controlled laboratory conditions or natural field conditions at different times of the year. Females exposed in a natural environment in September-October (short day) had significantly longer pre-oviposition periods than females exposed from November to March. The season in which the previous instar fed had no apparent effect on the engorgement or pre-oviposition periods of the females. Furthermore, artificial changes in photoperiod during and after female engorgement had no significant effects on pre-oviposition periods. It is tentatively concluded that the unfed female is the responsive stage to photoperiodic changes which induce diapause. Diapause could be terminated and oviposition induced by exposing females to a short period of chilling (18 degrees C for 48 h). It is concluded that a morphogenetic diapause mechanism exists in A.variegatum, which is probably induced by short day responses and terminated following rainfall and a concomitant decrease in soil temperature. The diapause, which occurs in females which fed early in the season, causes a delay in oviposition and therefore effectively synchronizes the life-cycle to ensure that eggs and larvae occur at a climatically favourable period. PMID- 2980188 TI - Pathological effects of Blastocrithidia triatomae (Trypanosomatidae) on the reduviid bugs Triatoma sordida, T. pallidipennis and Dipetalogaster maxima after coprophagic infection. AB - Developmental time and mortality in nymphs of the reduviid bugs Triatoma sordida (Stal), Triatoma pallidipennis (Stal) and Dipetalogaster maxima (Uhler) were studied in uninfected groups and in those infected with Blastocrithidia triatomae Cerisola et al. (Trypanosomatidae). In T.sordida and T.pallidipennis, major vectors of Chagas' disease in Brazil and Mexico respectively, infection with B.triatomae was associated with slight developmental retardations in the final instars, and increased mortality in the pre-adult instar. These effects were less evident in T.pallidipennis, but in this species a total infection rate of only 5 15% was achieved, compared with about 45% in T.sordida. In contrast, D.maxima was strongly affected by B.triatomae: nymphal development was retarded and complete mortality occurred in some groups exposed to infection, although not all bugs were infected. PMID- 2980189 TI - Ovitrap surveys of dengue vector mosquitoes in Chiang Mai, northern Thailand: seasonal shifts in relative abundance of Aedes albopictus and Ae. aegypti. AB - Aedes aegypti (L.) and Aedes albopictus (Skuse) were surveyed using ovitraps in residential areas in Chiang Mai, northern Thailand. Egg populations (both species inclusive) remained low in the dry season, but increased/decreased exponentially during the first/latter half of the rainy season, respectively. This seasonal pattern was similar to the seasonal distribution of dengue haemorrhagic fever cases in the area. During the dry season (November-March) Ae.aegypti was dominant in urban and indoor ovitraps. With onset of the rainy season in April, relative abundance of Ae.albopictus increased in rural and outdoor ovitraps. Ae.albopictus displaced Ae.aegypti in the latter half of the rainy season in the rural area. Possible mechanisms to account for this seasonal decline of Ae.aegypti and reciprocal fluctuations in relative abundance of Ae.albopictus are discussed in relation to food availability for larvae in container habitats. PMID- 2980190 TI - Observations on the susceptibility of Ctenocephalides felis (Siphonaptera: Pulicidae) to malathion and permethrin in Tanzania. AB - Laboratory-reared Ctenocephalides felis (Bouche) adults were tested with 0.5% malathion and 0.5% permethrin, using the standard WHO methods. After 24 h exposure to malathion (3.6 mg/cm2), 92% of the fleas died. The LT50 for malathion was approximately 8 h. Permethrin (0.45 mg/cm2) produced 100% mortality of exposed insects after 24 h while with a higher dose (0.9 mg/cm2) all fleas died after 8 h exposure. LT50 for the two doses of permethrin were 7.7 and 1.05 h, respectively. The failure of the diagnostic dose of malathion to kill 100% of the population was attributed to resistance. Permethrin is a suitable pesticide for controlling fleas of domestic animals in Tanzania. PMID- 2980191 TI - Anti-mosquito antibodies and their effects on feeding, fecundity and mortality of Aedes aegypti. AB - Anti-mosquito antibodies, produced in mice inoculated with mosquito homogenates or exposed to mosquito bites, reacted with a variety of mosquito antigens including muscle, gut, fat body and nervous tissue; those in anti-mosquito bite sera reacted solely with salivary glands. Mosquitoes fed on restrained immunized mice showed a significant increase in mortality correlated to both the titre and specificity of the anti-mosquito antibodies ingested. No changes in their fecundity or feeding success were noted. Mosquitoes exposed to unrestrained immunized mice or mosquito-bitten mice, however, showed a significant reduction in feeding success, possibly reflecting enhanced host grooming. PMID- 2980192 TI - Detection and localization of antibody ingested with a mosquito bloodmeal. AB - Mouse immunoglobulins were found to persist in Aedes aegypti (L.) mosquito bloodmeals for 2-3 days after ingestion. Immunoenzyme labelling revealed mouse antibody specifically bound to the mosquito midgut epithelium after ingestion; immunogold labelling of thin sections revealed mouse antibody within the cytoplasm of the microvilli on the midgut epithelium. Ingested mouse antibody was not conclusively demonstrated bound to tissues outside the gut, though antibody was detected in mosquito haemolymph using a sensitive ELISA test. Possible mechanisms by which antibody may cross mosquito gut barriers and actions that in vivo antibody:antigen reactions may have on these bloodfed insects are discussed. PMID- 2980193 TI - Survival and reproductive performance of female Glossina morsitans morsitans when maintained on the blood of different species of wild mammals. AB - A study was carried out to determine the effect on the reproductive performance of female Glossina morsitans morsitans Westwood when allowed to feed, in vitro, for 63 days on fresh defibrinated blood of buffalo, bushbuck, cattle, eland, oryx, warthog, waterbuck or wildebeest. There were marginal differences in the survival and reproductive performance between eight different groups of tsetse, 200 per group, when fed on the blood of these mammalian species. When allowed to feed for 14 consecutive days on the blood of buffalo, wildebeest or warthog, the mean number of feeds were 6.2 +/- 0.3, 6.5 +/- 0.3 and 6.3 +/- 0.3, respectively. The mean weight of the bloodmeal taken also did not differ significantly between these three groups. Whereas the protein patterns of the blood plasma of the above eight host animals were different, the protein patterns of the haemolymph from tsetse fed on the blood of these hosts were identical. It is thus concluded that the preference shown by tsetse for some mammalian species investigated here may not be based on any aspect of the nutritional value of their blood. PMID- 2980194 TI - Serologic diagnosis of Oestrus ovis (Diptera:Oestridae) in naturally infested sheep. AB - Sera from eighteen control sheep supposed to be free from parasitism by Oestrus ovis Linnaeus, 1761, and from 100 sheep raised in an enzootic area of O.ovis infestations were tested to detect anti-Oestrus antibodies by double immunodiffusion (DD) and indirect haemagglutination (IH) tests with somatic crude antigens from first (L1), second (L2) and third (L3) instar of O.ovis larvae. At necropsy, eighty-eight out of 100 sheep from the O.ovis infested area were found to be parasitized while the eighteen control ovines did not show Oestrus larvae. Examination of the sera from the parasitized sheep by DD showed positive results of 42% for L1, 59% for L2 and 18% for L3. Screening the sera with IH gave sensitivities of 100% for L1, 100% for L2 and 97.7% for L3. Sheep, naturally parasitized by gastrointestinal nematodes, presented no cross immune reactions in DD tests with the three larval stages of O.ovis or with L2 larvae in IH tests. PMID- 2980195 TI - Onchocerciasis transmission levels and Simulium damnosum complex biting activity at riverside and rice field sites in Sierra Leone. AB - Biting densities of the Simulium damnosum Theobald complex of blackflies and their levels of parasitism by Onchocerca volvulus (Leuckart) were compared at two ecologically contrasted sites in the Southern Province of Sierre Leone, West Africa: by the Tabe riverside close to Gbaiima village where onchocerciasis is hyperendemic (63.1% positive skin-snips) and in open rice fields 2 km from the mesoendemic (54.9%) village of Senehun. Using a combination of morphometric and enzyme electrophoresis techniques, the primary vector was found to belong to the S.sanctipauli subcomplex and was most probably S.soubrense 'B' (sensu Post, 1986). In the wet season biting activity took place throughout the day at the village, but had morning and evening peaks in the rice fields. Transmission of O.volvulus was detected at the village in all months except February-April, while in the rice fields it was detected only during September-November and May at one tenth the level of the village. There was a significant difference between the two sites in the proportion of parous flies carrying third stage larvae in the head, but not of other larval stages. PMID- 2980196 TI - Factors influencing silhouette trap captures of the blackfly Austrosimulium bancrofti in Queensland, Australia. AB - Three silhouette traps of novel design were used to investigate the trap finding ability of female Austrosimulium bancrofti (Taylor) (Diptera: Simuliidae) in southeast Queensland. The traps were 'upright', 'elongate' or 'square' in shape. Five highly significant factors: hour after sunrise, temperature, relative humidity, trap shape and distance between traps, were included in a model which generated fitted estimates of flies captured under defined conditions. The rate of increase and the level of maximum capture rate indicated that the elongate trap captured the highest numbers of blackflies. An interaction between hour after sunrise and temperature suggests a strong endogenous modification of trap finding activity. Numbers captured increased with increasing distance between traps and decreasing relative humidity. The importance of these results in understanding behaviour of A. bancrofti is discussed. PMID- 2980197 TI - Effects of salinity on the larvae of some Afrotropical anopheline mosquitoes. AB - Laboratory experiments showed that the larvae of some freshwater-breeding Afrotropical anopheline species exhibit various degrees of tolerance to 25% sea water. At 24 degrees C, survival rates from egg to fourth instar larva were significantly less in salt-water than in freshwater for four species: 39.5% versus 55.4% for Anopheles tenebrosus Donitz, 31.9% versus 86.2% for An. mousinhoi De Meillon & Pereira, 3.5% versus 67.9% for An. pharoensis Theobald and 3.9% versus 41.9% for An. quadriannulatus (Theobald). Conversely, An. merus Donitz showed a significantly better survival rate of 46.4% in 25% sea-water compared with 15.5% in freshwater. Simulated winter conditions did not significantly change these differential survival rates of An. merus. Effects of salinity on the anal papillae of third instar larvae are discussed. PMID- 2980198 TI - Leishmaniasis in the Jordan Valley. V. Dispersal characteristics of the sandfly Phlebotomus papatasi. AB - Several characteristics of dispersing and non-dispersing Phlebotomus papatasi (Scopoli) were quantified and compared. The majority of dispersing sandflies, trapped crossing fallow fields, were females (68.5% v. 51.1%); of the dispersing females, 55.4% were parous, 48.1% were inseminated and 11.6% were gravid. In the population of sandflies sampled exiting from burrows of the sand rat Psammomys obesus Cretschmar, these categories, respectively, represented 39%, 90% and 26% of the females examined. Leishmania promastigotes were found in 9% of females exiting from P. obesus burrows, and in 2.7% of the dispersing females. The anthrone test established that the reason for activity of gravid females is sugar feeding. These females do not disperse and are suitable targets for future control measures. PMID- 2980199 TI - Natural hybridization between Simulium sanctipauli and S. sirbanum, two sibling species of the S. damnosum complex. PMID- 2980200 TI - Dispersal flight by Triatoma infestans in an arid area of Argentina. PMID- 2980201 TI - Identification of a nodD-dependent locus in the Rhizobium strain NGR234 activated by phenolic factors secreted by soybeans and other legumes. AB - Transfer of the strain NGR234nodD 1 gene into the narrow host range R. trifolii strain ANU843 on either a 6.7-kb HindIII or 17-kb XhoI fragment broadens the host range of this bacterium to include the tropical legumes Vigna unguiculata, Glycine ussuriensis, Leucaena leucocephala, and siratro (Macroptilium atropurpureum). Contrary to previous data (Bassam et al. 1986), mutagenesis of the 17-kb XhoI fragment with a mini-Mu lac transposon (Mu dII1734) showed that a functional nodD 1 gene was essential for extended host range. Gene expression studies using both Mu dII1734 fusions and a promoter-cloning vector indicated that several loci, including the nodD 1 gene, are constitutively expressed. No evidence was found for regulation of the strain NGR234 nodD 1 gene by its product. Another locus nod-81, was induced only in the presence of exudates from various plant species, including soybean (Glycine max). Whereas the expression of nod-81 was dependent on the presence of a functional nodD 1 gene product, a regulatory nod-box DNA sequence was not detected 5' to this gene by using available oligonucleotide hybridization probes. The nod-81 locus was induced by genistein, daidzein, naringenin, and coumestrol from both cotyledon and root tissue of freshly germinated soybean seedlings. A broad spectrum of commercially available phenolic compounds stimulated induction of the nod-81 locus, including some that antagonize nod gene induction in other Rhizobium species. The nodD 1 gene product from strain NGR234 was shown to determine the spectrum of compounds that induce nod-81 expression. PMID- 2980202 TI - Pathogenicity regulation by satellite RNAs of cucumber mosaic virus: minor nucleotide sequence changes alter host responses. AB - Six satellite RNAs of cucumber mosaic virus (CMV) could be differentiated on the basis of symptom expression they elicit in tomato and tobacco, and all but two could be differentiated by gel electrophoretic migration. Three of the satellite RNAs (B2-sat, G-sat, and WL1-sat RNA) ameliorated the symptoms induced by CMV on tomato, whereas three others (B1-sat, B3-sat, and WL2-sat RNA) induced chlorosis on tomato, the extent and nature of which was CMV-strain dependent. By contrast, B2-sat RNA induced chlorosis in tobacco, whereas WL1-sat and G-sat RNAs did not. Thus, the symptoms observed were dependent on the host species, the particular satellite RNA, and also the strain of helper virus, suggesting that a complex association of at least three factors is involved in symptom elicitation. Comparisons of the nucleotide sequences of pairs of satellite RNAs inducing the various chlorotic responses suggest that only a few nucleotide changes in specific domains are required for the elicitation of different host responses. PMID- 2980203 TI - Ultrastructural computerized morphometry of platelets in chronic myelogenous leukemia making use of ultra-thin sections and freeze-fracture procedure. A preliminary approach. AB - Biochemical and morphological data have shown that the circulating platelets are deeply altered in patients with chronic myelogenous leukemia. In this report we describe the results of ultrastructural morphometry performed by means of a computerized device of the platelet shape (ultra-thin sectioned platelets) and of the platelet plasma-membrane (freeze-fractured platelets). Platelets appeared deeply modified: reduced mean platelet area and perimeter, increased surface density of the openings of the surface connected canalicular system, abnormal features of the platelet cytoplasm and some aspects of platelets joined together even if with heterogeneity in these findings did appear from patient to patient. PMID- 2980204 TI - A light and electron microscopic study of the preoptic nucleus of the grass frog (Rana pipiens), under normal conditions and following transection of the preoptico-neurohypophysial tract. AB - Degenerative and regenerative processes occur in the preoptic neurons following transection of the preoptico-neurohypophysial tract. Three types of responses after transection were observed: affected, recovered, and degenerated neurons. However, transection of the tract did not stop the synthesis of neurosecretory granulated vesicles. The affected neurosecretory neurons showed nuclear changes, increased number of Golgi complexes, and dilated cisternae of rER, as well as, an increased number of dense bodies. The recovered neurosecretory neurons contained long non-dilated cisternae of rER which were organized in a concentric manner. Also seen were large nuclei with evenly distributed chromatin, less active Golgi complexes, and vesicles. The degenerated neurosecretory neurons exhibit pyknotic nuclei, a net of dilated cisternae of rER, dense bodies, and electron dense cytoplasm. PMID- 2980205 TI - Retinal epithelial fine structure in the vervet monkey (Cercopithecus aethiops). AB - The morphology of the retinal pigment epithelium (RPE) and closely associated Bruch's membrane (complexus basalis) and choriocapillaris have been investigated by electron microscopy in the vervet monkey (Cercopithecus aethiops). The RPE is composed of a single layer of cuboidal cells joined laterally by apically-located junctional complexes. Basally (sclerally) these cells display numerous infoldings while apically (vitreally) abundant processes enclose and interdigitate with rod outer segments. Internally the large vesicular nucleus is centrally located and smooth endoplasmic reticulum, mitochondria and lysosome-like bodies, are plentiful. Rough endoplasmic reticulum, polysomes and melanosomes while present are not abundant. Phagosomes of outer segment discs are noted in various stages of uptake and degradation. The choriocapillaris is highly fenestrated over large areas. Bruch's membrane shows the typical pentalaminate structure noted in other mammalian species without a tapetum lucidum. PMID- 2980206 TI - Effects of orthodontic tooth movement on the Alcian blue staining patterns of rat alveolar bone: an histochemical study. AB - There is little information available concerning the effects of orthodontic forces on glycosaminoglycans (GAG) of alveolar bone. The present study identifies changes in Alcian blue staining intensity in rat alveolar bone undergoing resorption resulting from a heavy (25g) tipping force applied to the adjacent teeth by a separating spring. One day after force application, bone from treated animals (internal control and experimental sides) demonstrated more intense staining with Alcian blue, pH 1.0 (p less than 0.005) and pH 2.5 (p less than 0.05) than external controls (untreated animals). By day 3, the intensity of Alcian blue staining of treated alveolar bone was similar to untreated. Chondroitinase AC, ABC and testicular hyaluronidase predigestion did not completely block the staining reaction, suggesting that both GAG and noncollagenous proteins were demonstrated. Mean cross-sectional areas of the interdental septum of the experimental side were nearly 44% less than that of the internal control side after 3 days and nearly 62% less after 5 days. The study suggested that alterations in bone GAG levels occurred prior to tooth movement as histochemical changes occurred after force application but before initiation of significant septal resorption. A precise appraisal of the types of macromolecules effected awaits future biochemical analysis. The results of the present work strongly suggest the use of an external control group for future studies, as Alcian blue staining reactions of the internal control side of treated animals were not similar to those of external controls. PMID- 2980207 TI - Ultrastructural pathologic changes of rat extraembryonic visceral endodermal cells exposed to teratogenic antibodies in vivo. AB - It has been well established that certain heterologous tissue antibodies may induce abnormal embryonic development when injected into pregnant rodents during the organogenetic period. It has been postulated that these antibodies indirectly cause embryopathy by interfering with the normal functions of the yolk-sac placenta. The exact mechanism whereby these antibodies may induce placental pathology is not known. Specific teratogenic antibodies against a homogeneous rat kidney glycoprotein or a visceral yolk-sac glycoprotein antigen were injected intraperitoneally into 9th day pregnant rats. Electron microscopic examinations of the extraembryonic visceral endodermal cells of the egg cylinder were performed at 4, 6, 9, and 24 hours after the administration of the teratogenic antibodies. Control animals were injected with normal rabbit serum proteins. Extraembryonic visceral endodermal cells were similarly processed and examined as the experimental groups. The results seemed to indicate that the teratogenic antibodies induced increased autophagocytosis and morphologic changes associated with the phagolysosomes (secondary lysosomes) within the extraembryonic visceral endodermal cells at 9 hours following antibody administration. After 24 hours there was an apparent reduction or a complete disappearance of the supranuclear phagolysosome-like and lysosome-like structures, and the appearance of many large and small electron lucent vacuoles containing finely granular materials. Similar ultrastructural pathology was not observed in the 4 and 6 hour experimental and all of the control groups of animals. No other obvious intracellular or intercellular changes were observed in all of the experimental groups. Although the exact mechanism whereby the teratogenic antibodies may induce pathologic changes in the extraembryonic visceral endodermal cells remains to be determined, the present ultrastructural study demonstrated, for the first time, that teratogenic antibodies induced abnormal pathology in the extraembryonic visceral endodermal cells during the critical period of organogenesis. PMID- 2980208 TI - Hepatitis B core and surface antigens and delta agent in chronic liver disease in Kuwait. AB - Hepatitis B surface antigen was detected immunohistochemically in 25 out of 85 liver biopsies (29.4%) of chronic liver disease. Core antigen was also demonstrated in 9 of the 25 Hepatitis HBs Ag positive biopsies (36%). Delta agent however, was found in only one case of HBs positive chronic active hepatitis. The number of hepatocytes staining positively for HBc antigen was greater in those biopsies with the strongest staining for HBs antigen. The only case of chronic active hepatitis positive for delta agent showed that the positive staining was confined to the nuclei of few hepatocytes. The routine histology showed chronic active hepatitis with a moderate degree of inflammation. The present results confirm our previous reports that almost one third of chronic liver disease in Kuwait is associated with hepatitis B infection. Previous serological studies suggest that delta agent infection is also common; however, the present study suggest that delta agent may be a transient, and not a major, contributing factor in the progression of liver disease. PMID- 2980209 TI - Squamous and glandular differentiation in urothelial bladder carcinomas. Histopathology, histochemistry and immunohistochemical expression of carcinoembryonic antigen. AB - This paper reports the immunochemical expression of carcinoembryonic antigen in the squamous, glandular and mixed differentiation areas observed in 190 urothelial urinary bladder carcinomas. The antigen was found to occur in 75% and 81% of all papillary and solid infiltrating carcinomas, respectively. The use of the immunohistochemical determination of CEA in improving the morphological examination of these differentiation areas and their scant presence in the prognosis of urothelial carcinomas involving squamous and/or glandular focal differentiation are discussed. PMID- 2980210 TI - Immunophenotypic characterization of primary and secondary lymphoid follicles. AB - The need for an immunophenotypical referential framework relative to lymphoid follicle has led us to apply a panel of monoclonal and polyclonal antibodies, by means of a sensitive immunostaining method. Lymphoid follicle is an immunophenotypically complex structure made up of three lymphoid populations (B, being its bulk, and a few T and NK cells), dendritic reticulum cells (DRCs) and Flemming's macrophages. Follicular B population is To 15 +, B1+, OKB 7 +, HLA-DR + and C3bR +. In secondary follicles there are differential characteristic reactivities for each topographic compartment: Mantle zone is positive for OKB 2 and surface IgM (sIgM) and IgD (sIgD); germinal center (GC) clear zone (with centrocytic predominance) for OKT 9, sIgM and weakly for OKB 2; and GC dark zone (with centroblastic predominance) only for OKT 9. In sections, OKT 10 allows one to see immunoblasts and plasma cells, the latter being with lymphoplasmacytoid cells the only intracytoplasmic immunoglobulin holders. 10% of GC lymphocytes are T cells, almost exclusively T-helper (Leu 3a +). Another 10% to 15% of lymphoid cells are Leu 7 (HNK-1) +. In histological sections, DRCs are specifically marked with R4/23 and Flemming's macrophages with anti-alpha1-antitrypsin and anti alpha1-antichymotrypsin antibodies, both populations being negative to OKM 1 and OKM 5. PMID- 2980211 TI - Distribution of ferric iron in larval lampreys, Petromyzon marinus L. AB - The distribution and abundance of ferric iron in larval lampreys (Petromyzon marinus L.) were investigated using light microscopy and the Prussian blue stain. Animals from various watersheds contained different concentrations of iron, although the sites of deposition were the same for all animals. A major portion of iron is within adipose tissue, while the liver, and cartilage contain predominantly low to trace amounts of iron, respectively. Iron is associated with fibrous connective tissue in several places in the body, and this association may have particular significance in the inner ear. Iron is also located in cells of the meninges. The presence of iron in the epithelial cells of the posterior intestine may reflect elimination of the metal through the extrusion of iron loaded cells into the intestinal lumen. Iron within mucous cells of the epidermis, suggest elimination of iron during mucous secretion. Iron-loaded cells of bipolar shape are also present in the epidermis, but are particularly prominent around the branchiopore. Low concentrations of iron are observed within in melanin-containing macrophages (melano-macrophages) in regions of iron absorption, erythrophagocytosis, and haemopoiesis. High levels of iron in the epithelia and lumina of pronephric tubules are concomitant with degeneration of this organ. These data are evidence of the wide spread distribution of iron in lamprey tissues and additional evidence for the potential value of lampreys for the study of iron metabolism in vertebrates. PMID- 2980213 TI - Immunocytochemical localization of parathyroid hormone in rabbit parathyroid glands. AB - Immunocytochemical localization of parathyroid hormone was examined in the rabbit parathyroid gland by means of protein A-gold technique. Protein A-gold particles were observed on the secretory granules and the large secretory granules thought to be storage granules. No protein A-gold particles were observed on cisternae of the endoplasmic reticulum and the Golgi apparatus. PMID- 2980212 TI - Immunohistochemical study of Warthin's tumour with special regard to the germinal centre. AB - In order to analyze the role of the lymphoid stroma of Warthin's tumour, the author made an immunohistochemical study especially focussing on the germinal centre (GC). In the GCs, IgM and IgE were much more clearly observed in a lace like network than other heavy chains, and also could be detected electron microscopically on the surface of GC cells and follicular dendritic cells (FDCs). Plasma cells scattered beneath the epithelial component were mainly positive for IgG and IgA. IgG-positive plasma cells were more predominant than IgA plasma cells. Among the cases examined, there was a significant difference in the number of IgE-positive GC and plasma cells. In the GCs five complement components (C1g, C4, C3c, C3d, C5) and complement receptors (C3bR and C3dR) were positive, displaying a lace-like pattern, which were proved, electron microscopically, to coincide with the surface of FDCs and GC cells. IgE-positive GCs showed the same result. DRC1, which specifically reacts to FDC-membrane, was located only in GCs, and electron microscope also revealed positive findings on the surface of FDCs. The above mentioned findings in the GC of Warthin's tumour were similar to those of lymph nodes except for the frequent distribution of IgE. Salivary amylase was seen in the GC on rare occasions, but was not positive on the surface of GC cells and FDCs. It is suggested that FDCs play an important role in immunological behaviour with complement and complement receptors in the GC of Warthin's tumour. PMID- 2980214 TI - Neuron specific enolase-positive breast carcinomas. AB - Ninety-eight patients treated for breast carcinomas were followed from 54 to 75 months after primary diagnosis. All had undergone a modified radical mastectomy with removal of axillary lymph nodes. 36 breast carcinomas were NSE-positive and 62 were negative. NSE-positive tumours were significantly more frequently estrogen receptor-positive than the NSE-negative tumours, and the estrogen receptor values were higher in the NSE-positive groups. Patients with NSE positive tumours and patients with NSE-negative tumours did not differ with regard to presence of lymph node metastases at the time of primary surgery. However, the study showed that patients with NSE-positive tumours had a tendency towards more lymph node metastases after primary surgical intervention, but a better outcome than patients with NSE-negative tumours and metastases. This study, with a 5-year follow up, failed to demonstrate any major prognostic significance of immunostaining for NSE. PMID- 2980215 TI - Fine structure of the receptors at the myotendinous junction of human extraocular muscles. AB - The myotendinous junction of the human extraocular muscles was studied by electron microscopy. Some peculiar receptorial structures have been found in the majority of the samples examined. These structures are very small and consist of 1) the terminal portion of one muscle fibre, 2) the tendon into which it inserts and 3), within the tendon, a rich nerve arborization, whose branches are always very close to the muscle component. Only one discontinuous layer, made up of flat cells, which lack a basal lamina and often show pinocytotic vesicles, encapsules every musculo-tendinous complex. The tendinous component consists of amorphous ground substance of different electron density, of collagen and elastic fibres and is divided in compartments by ramified cells, which make an inner capsular like covering to the nerve fibres. Three types of afferent nerve endings can be identified. One type is usually more frequent than the others, possesses a large number of neurotubules and neurofilaments and few mitochondria and is always surrounded by a Schwann cell which forms finger-like processes penetrating into the axoplasm. The second type is only partially enveloped by the Schwann cell. The axoplasm is devoid of neurotubules and contains few neurofilaments, several mitochondria and groups of small clear vesicles placed in the areas uncovered by the glial sheath. The third one is completely surrounded by the Schwann cell, but is devoid of neurotubules and neurofilaments and full of mitochondria. These morphological features correspond well with the probable role of these receptorial structures, which is to ensure very exact and precise ocular movements. PMID- 2980216 TI - Intrinsic and extrinsic innervation of the amphibians esophagic myenteric plexuses. AB - The innervation of Rana ridibunda esophagus myenteric plexuses has been studied by the following methods: demonstration of cholinesterase activity; FIF method for catecholamines; immunohistochemistry for VIP, SP and SOM, and conventional electron microscopy. The cholinergic innervation is important in the esophagus wall where, in addition to the well known extrinsic component, there is a rich intrinsic plexus with cells and fibres widely distributed. The esophagus, together with the intestine, are the Rana gut portions where the adrenergic component is more broadly expressed. The adrenergic innervation seems to be almost entirely of extrinsic origin. We have shown that, for the tested peptides, there is an intrinsic innervation represented by VIP, SP and SOM like plexuses. We do not discard nonetheless an extrinsic component. The ultrastructure reveals the morphological characteristics of the enteric neurons as well as the fine inter-relationships between the nervous elements and the functional components of the esophagic wall. PMID- 2980217 TI - Pulmonary changes in rats following administration of 3-methylindole in cremophore EL. AB - 3-methylindole (3-MI) dissolved in the lipophilic carrier Cremophore EL was administered intraperitoneally to male, twelve-week-old Sprague-Dawley rats. Gross and histopathologic changes in the lungs were studied using light microscopy at three time-periods following administration: 16, 24, and 46 hours. Both 3-MI and Cremophore caused changes in bronchiolar epithelium at 16 hours. By 46 hours, Cremophore-injected rats showed no effects of the carrier; whereas, 3 MI rats showed severe lung changes characterized by airway epithelial and pulmonary vascular endothelial necrosis and sloughing, cellular infiltration by lymphocytes and macrophages, perivascular edema, alveolar edema, and lymph stasis. Grossly, the controls showed no effect of the carrier and none died during the studies. In contrast, 3-MI injected rats quickly became lethargic and displayed tachypnea, anorexia, and progressive respiratory distress. Two of five 3-MI rats in the final group died just prior to 46 hours. All of this group had grossly congested lungs and marked pleural effusion. The lesions and time course showed similarities to those observed in ruminants and mice. We conclude that 3 MI in Cremophore causes an acute progressive pneumonitis in rats and suggest that the rats may be a suitable model for 3-MI-induced and similar toxic lung diseases in domestic animals and people. PMID- 2980218 TI - Ultrastructural changes of the renal cortex after septic shock in rats. AB - The purpose of this study was to observe the ultrastructural changes of the renal cortex that occur in septic shock. Twenty male Wistar rats were used. They were divided into 4 groups of five animals each one. A septic shock was caused by intraperitoneal injection of 5 x 10(6) living E. Coli, in a dose of 2ml/100gr B. W., in four rats of each group. The fifth rat of each group was used as control, and was injected by an equal volume of normal saline. A small segment from the right kidney was excised from every rat of each group, respectively 1, 2, 4 and 6 hours after the onset of the shock. Morphologically, alterations of the endothelium of the peritubular and glomerular capillaries were found. The endothelial layer was thick with many cytoplasmic folds. In the glomeruli, the basal lamina as well as the overlying epithelial podocytes and their foot processes were swollen. The urinary space contained erythrocytes and cell debris. Lesions of the proximal and distal tubule epithelium were also observed. The basal infoldings of the cell membrane disappeared in many epithelial cells of the proximal tubule and the brush border showed varying degrees of disruption. Within the tubular lumen much cellular debris was found. The epithelium of the distal tubule was thinner in some portions and in others it was swollen. The lesions were focal during the first hour after the onset of shock, while at the fourth and sixth hours they were most extensive. In conclusion, the ultrastructural findings after septic shock were non-specific and indicated cellular injury. PMID- 2980219 TI - Morphological study of the rat pituitary follicle stimulating hormone cells after alternate day treatment with luteinizing hormone-releasing hormone (LHRH). AB - The action of the luteinizing hormone-releasing hormone (LHRH) on the hypophyseal gonadotrophins has either an activatory or inhibitory effect depending on the doses administered or on the treatment followed. Both factors can induce a different response in the two hormones. In this work, the effect after the administration of 8 doses (40 micrograms/day) of LHRH at intervals of 48 hours, on the serum levels of follicle stimulating hormone (FSH), as well as the numerical density, distribution, intensity of staining and morphometrical parameters of the cells which react against the anti-FSH serum, are assessed. It has been found that with the treatment an increase of FSH serum levels, without modification in the number of immunoreactive cells, but a clear increase in the lightly stained cells, is produced. The distribution of the reactive cells, uniform in normal animals, shows a large numerical density in the dorsal and posterior hypophyseal areas in the treated animals. No change was observed in the nuclear and cellular areas between the different groups. PMID- 2980220 TI - Ultrastructural observation on the rat supraoptic neurons following acute operative stress. AB - The response of the SON to various forms of stress is well documented. However, the effect of operative stress, which is a common and important clinical event requiring the mediation of vasopressin, has largely escaped attention. The present report describes the ultrastructural changes in the neurons of the caudal (retrochiasmatic) part of the SON following a deepseated linear incision on the dorsum of the rat. The observations were confirmed to the first forty-eight hours after trauma. At 24-hours post-operatively, a marked depletion of the neurosecretory granules was observed. This was associated with a proliferation of the granular and smooth endoplasmic reticulum, Golgi cisternae and ribosomes. A few of the neurosecretory granules were seen to lie in the close vicinity of the Golgi complexes. At 48-hours after trauma, these features persisted. In addition, an accumulation of neurosecretory granules was conspicuous in some axon pre terminals. From the above findings, it is suggested that an increased demand for vasopressin during the early postoperative period is met by the supraoptic neurons by a liberation of their neurosecretory contents. An attempt at replenishment of the latter is evidenced by a proliferation in the membrane components and ribosomes. The pooling of neurosecretory granules in occasional axon pre-terminals may indicate an imbalance in the synthesis-secretion coupling of vasopressin. PMID- 2980221 TI - The origin and differentiation of adrenocortical cells in rats with portacaval shunt. A structural and ultrastructural study. AB - The formation of adrenocortical cells in the rat was studied by light and electron microscopy in an experimental model, namely portacaval shunt (P.C.S.), in which strong hyperplasia of the cells of the capsular region occurs. The results of this study indicate that in physiological conditions at the level of the adrenal gland capsule some epithelial cells, morphologically distinguishable as dark and clear cells, are found which can be interpreted as precursors of adrenocortical cells. From observations of intermediate forms between capsular precursors and mature adrenocortical cells, which are found in high numbers following P.C.S., it seems that the dark precursors give rise to cells of the zona glomerulosa and the clear precursors evolve into cells of the zona intermedia, which are to be considered as the starting point for the formation of cells of the zona fasciculata. PMID- 2980222 TI - Alkaline phosphatase activity of the IVth ventricular choroidal epithelium of rats during embryonic and neonatal development. AB - The cytochemical localization of alkaline phosphatase (AlPase) activity in the developing IVth ventricular choroidal epithelium was investigated in embryonic and neonatal rats. During the initial development of the choroidal primodium the flattened and/or cuboidal epithelial cells of the ventricular roof were changed to columnar cells with well-developed microvilli and apical tight junctions. When compared to AlPase activity on the lateral plasma membranes of the surrounding ependymal cells, these columnar cells of the choroidal primodium revealed activity on the lateral and luminal plasma membranes, but no activity was found on the basal surface of these cells. On the other hand, the epithelial cells in the neonatal choroid plexus showed a continuous morphological alteration from columnar cells with short microvilli to mature cuboidal cells with numerous long microvilli. AlPase activity in immature columnar cells was observed on all plasma membranes, except for the apical junctional area of the lateral surface. With maturing of the choroidal epithelial cells, the activity appeared to be eliminated from the lateral and luminal plasma membranes of the cuboidal cells, and mature choroidal epithelial cells showed activity on the basal surface only. These findings suggest that AlPase may play an important role in the membrane activity of epithelial cells differentiating between the primitive epithelial cells of the ventricular roof and the mature choroidal epithelial cells. PMID- 2980223 TI - Gap-like junctions between neurons and glia in the superior colliculus of mammals. AB - The neuron cell bodies and glia cells are separated by a narrow intercellular cleft of 200 A. We present here evidence that perikarya neuronal and perineuronal glia of the superior colliculus of the Oryctolagus Cuniculus communicate through gap-like junctions according to ultrastructural parameters. These junctions would mean some degree of electronic coupling between neuron cell bodies and perineuronal glia cells. PMID- 2980224 TI - Occurrence of lipofuscin pigment granules and "dense microspheres" in the spinal cord of young cats treated with beta,beta'-iminodipropionitrile (IDPN). AB - Spinal cords of cats treated with the neurotoxic compound beta,beta' iminodipropionitrile (IDPN) were observed to contain rounded homogeneous bodies, 1-12 microns in diameter, termed "dense microspheres" (DMS). These bodies, absent in control animals, were consistently found only in the ventral horns. No relationship with blood vessels was evident. When stained with PAS and a modified von Kossa's silver nitrate technique, DMS remained negative, showing only very slight metachromasia in some toluidine blue-stained sections. They were consistently acidophilic as evidenced by destaining and differentiation investigations. DMS were observed more frequently in the proximity of nerve cell bodies or closely adjacent to dendrites and their location was mainly extracytoplasmic; with the electron microscope, however, some DMS were also found in glial processes. Rounded osmiophilic bodies, 0.1-0.8 microns in diameter, were noticed in mitochondria of both neurons and glial cells; however, whether they were special forms of DMS or different inclusions was not assessed. Both intra- and extracytoplasmic DMS were similar in ultrastructure, appearing as single membrane-bound spherical or pear-shaped bodies containing a cottony or finely granular matrix. Additionally, both perikaryon and processes of large motoneurons were found to contain pigment granules identified as lipofuscin, which seemed to increase in number and to spread centrifugally in the processes in correlation with duration of the intoxication and size of axonal swellings induced by IDPN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980225 TI - Colchicine causes intrasomatic neurofilament bundles in the mesencephalic trigeminal nucleus and intradendritic bundles in other brain regions. AB - The effect of a single intracerebroventricular injection of colchicine on the distribution of organelles in neurons of the mesencephalic nucleus of the trigeminal nerve, the inferior colliculus and the deep cerebellar nuclei was studied. In the mesencephalic nucleus of the trigeminal nerve colchicine produced a dramatic accumulation of neurofilament bundles in the soma of these neurons and did not produce a reduction in the number of lysosomes. In other neuronal populations studied, colchicine produced neurofilament bundles in the dendrites and a reduction of lysosomes from the soma of neurons. PMID- 2980226 TI - Blood-brain barrier (BBB). Review from morphological aspect. PMID- 2980227 TI - Maternal smoking during pregnancy affects neuroendocrine cells in the neonate hamster lung. AB - Primigravid Syrian golden hamsters were exposed in a Walton smoking machine to the smoke from either weak or strong cigarettes for 10 minute periods, 4 times a day from the 3rd to 14th (2nd last) day of pregnancy. Control hamsters were either similarly restrained in a Walton machine equipped with an unlit cigarette, or were not placed in the machine or restrained. Examination of the progeny in the first 6 days of life showed changes in density indices of grouped pulmonary neuroendocrine (NE) cells (neuroepithelial bodies, NEB) that were related to in utero exposure to maternal smoking. Argyrophil NEB were more numerous, larger, and contained more cells at birth among neonates whose mothers smoked the strong cigarette (2.45 mg nicotine and 36.8 mg tar) during pregnancy. This suggests a dose-related effect as the weak cigarette (0.37 mg nicotine and 33.8 mg tar) group did not show such changes. However, some of the changes described did not last through 3 or 6 days of age. The stress resulting from restraint alone also appeared to increase argyrophil NEB indices. Lung tissue volume fraction was increased in the weak cigarette group over all other groups at birth and 3 days; this suggests that low nicotine has the strongest pharmacological effect on lung tissue growth. The medial thickness of pulmonary arterioles was unchanged by either treatment; this provides morphometric evidence that chronic pulmonary hypertension was not present. We could not determine whether the increased NEB indices were caused by increased stainability, by activation of resident reserve cells, or by actual mitosis. PMID- 2980228 TI - Effects of isoproterenol on the fine structure of the hamster parathyroid gland. AB - Ultrastructural changes of the parathyroid glands of isoproterenol-treated golden hamsters were investigated. Many chief cells in the parathyroid glands after 5 and 10 minutes of administration of isoproterenol contain well-developed Golgi complexes and granular endoplasmic reticulum, numerous prosecretory granules, and many secretory granules in the peripheral cytoplasm as compared with the control animals. Many chief cells in the parathyroid glands after 1, 3, 6 and 12 hours of administration have poorly-developed Golgi complexes, granular endoplasmic reticulum, many secretory granules and numerous lipid droplets as compared with the control animals. The morphology of the parathyroid gland after 30 minutes and 24 hours of administration resembles that of the control animals. It is considered that isoproterenol affects the secretory activity of the parathyroid gland. PMID- 2980229 TI - Metaplasia of the parietal layer of Bowman's capsule in the human kidney: incidence in alcoholic liver disease and hypertension. AB - This report is the second of two surveys to determine the incidence of metaplasia of Bowman's parietal epithelium in the human kidney. Human kidney sections obtained at autopsy at the Department of Pathology, University of Texas Medical Branch, Galveston, Texas, were examined with the light microscope. The kidneys were fixed in neutral formalin, sectioned at 6 microns and stained with hematoxylin and eosin. Autopsy records were consulted after kidney section examination to determine if there was any correlation between clinical disease, histopathological changes in organ systems and metaplasia of Bowman's capsule. The kidney sections represented both sexes in 8 age groups, from less than one year to 80 years. A total of 174 kidneys, representing 174 individuals, were evaluated. One hundred renal corpuscles were counted per section and the parietal layer of Bowman's capsule was classified as normal (squamous) or metaplastic (cuboidal). Of the 174 kidneys examined, 137 (79%)--79 male and 58 female--had metaplasia of Bowman's capsule. On the average, in the kidneys with the lesion, 6% of the renal corpuscles had metaplasia of Bowman's parietal layer. The lesion was present in both sexes in all age groups. The autopsy records revealed that metaplasia of Bowman's parietal epithelium was usually present with hepatic fatty changes and/or congestion. Alcoholic liver disease and hypertension represented the most frequent clinical diseases in the sample; these conditions had the highest incidence of metaplasia. Twenty-six of the 174 kidney samples were from individuals with alcoholic liver disease, all of whom had metaplasia of Bowman's capsule.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980230 TI - An ultrastructural stereologic study of aldosterone secreting adrenal adenomas and of adjacent zona glomerulosa. AB - The ultrastructure of four aldosterone secreting adenomas and of the adjacent zona glomerulosa has been described by the use of stereological techniques. Adenomatous cells (about 2800 microns 3 in volume) invariably displayed a striking abundance of lipid droplets, which occupied about 30% of the cytoplasm. Mitochondria prevalently contained tubulo-lamellar or lamellar cristae, but some cells exhibited organelles with vesicular cristae. Smooth endoplasmic reticulum (SER) was not very abundant. Small lipofuscin-pigment granules were frequently seen and in a few cells they were exceedingly numerous. Zona glomerulosa cells were smaller (about 950 microns 3 in volume) and possessed mitochondria with typical tubulo-lamellar cristae, a plentiful SER and few lipid droplets. They showed the ultrastructural features of elements actively engaged in steroid synthesis. The possible origin of aldosteronoma cells from the zona glomerulosa is discussed. PMID- 2980231 TI - The influence of matrix components on the morphological differentiation of a proliferating hepatocyte line from liver of newborn mice. AB - The differentiation behaviour of a liver epithelial cell line of the newborn mouse cultured on various matrix components (Thermanox pure, Thermanox coated with ECM, dried collagen type I and type II, wet collagen type I and type III and on floating collagen) was investigated by electron microscopy. Only during the last few days of pregnancy and up to day 9 p.p. could these cells be isolated using a very delicate method. The cells were smaller than differentiated hepatocytes and proliferated spontaneously. They resembled the so-called oval liver cells. On Thermanox pure or Thermanox coated with ECM, dried collagen type I or type II a confluent monolayer developed after about 6 days that consisted of rather flat extended cells which were characterized by short contacts and the absence of any morphological indications of differentiation. On wet collagen the extension area was smaller and the cells were taller. The length of the contact area and the number and size of gap junctions and cell organelles increased. On floating collagen multi-layered aggregates of polygonal cells developed that were characterized by extended cell contacts, bile capillary-like structures and highly developed cell organelles, especially rough endoplasmic reticulum. Since differentiation processes can be demonstrated ultrastructurally only on wet collagen, especially on floating collagen, the chemical composition of the substrate and a specific matrix-cell interaction cannot be the only triggering factor. It is assumed that mechanical properties of the substrate, e.g. plasticity, are involved. The change in the shape of the cell, the prolongation or intensification of the cell contact and the adaptation of the cytoskeleton might play a decisive role in this connection. PMID- 2980232 TI - Pituitary growth hormone secretory granules in streptozotocin-induced diabetic rats. AB - The ultrastructural features of growth hormone (GH) secretory granules were observed by immuno-gold procedure in the anterior pituitary glands of streptozotocin (STZ)-induced diabetic rats. The diabetic state of rats induced by one injection of STZ was severely advanced at 10 months after the injection. The cytoplasmic organella were revealed to be poorly developed in GH-cells of the diabetic rats. The size of immunoreactive GH-granules of 10-months-old diabetic rats was smaller than that of sham-controls. The insulin therapy in the diabetic rats resulted in a considerable improvement in relation to the changes seen in the GH-cells, but the therapy did not return the profile of the granule size distribution to that of the control group. PMID- 2980233 TI - An immunocytochemical method for assaying oestrogen receptors in breast cancers. A comparison with the steroid binding assay. AB - The presence of oestrogen receptors was studied in 105 human breast carcinomas using monoclonal antibodies (Abbott ER-ICA kit). The oestrogen receptors of neoplastic cells were semiquantitatively measured and correlations were made to receptor values determined by a dextran-coated charcoal (DCC) steroid binding assay and to histological grade. Immunoreactive cells were found in about 2/3 of the tumours. Usually only a fraction of the cells within each tumour were immunoreactive, and the staining intensity varied among different cells. In general, well differentiated tumours had a greater proportion of immunoreactive cells than poorly differentiated ones. In most cases (65/98) a good agreement was found between the ER-ICA method and the DCC assay. However, in 33 cases discrepancies were demonstrated. PMID- 2980234 TI - Previously unreported "hemidesmosomal junctions" between folliculo-stellate cells and pituitary adenoma cells. AB - Thirty-eight non-functioning pituitary adenomas were ultrastructurally investigated with particular attention to the Folliculo-Stellate (FS) cells. A large number of FS cells were found in four cases, one of which disclosed a new type of intercellular junction between FS cells and surrounding adenoma cells. These junctions were characterized by 1) the presence of plasmalemmal attachment plaques only in FS cells, 2) the cytoplasmic filaments assembling in parallel to the attachment plaques, 3) the parallel plasma membranes being separated by the intercellular amorphous material and 4) the intercellular space of approximately 25 nm width. They were similar to hemidesmosomes, but were quite different from hemidesmosome-like intercellular specializations which have been described in the normal meninges and human meningiomas. Accordingly, we designated these new junctions as "hemidesmosomal junctions" which appeared to be one of the ultrastructural features characterizing FS cells. PMID- 2980235 TI - Structural and ultrastructural modifications following transurethral injection of teflon paste. AB - Structural and ultrastructural tissue modifications caused by polytetrafluoroethylene (teflon) paste injection in incontinent male patients have been studied. The most considerable observations are: a diffuse interstitial splitting of teflon paste in particles of about 3-100 microns. Given that the tissue reaction is limited to a modest infiltration of lymphocytes and monocytes and a slight increase in the collagen fibres, it is concluded that the teflon paste injection is well tolerated by the tissues. PMID- 2980236 TI - Proteoglycans of alveolar bone of diabetic and non-diabetic mice: a histochemical study. AB - The effect of diabetes mellitus on the interdental alveolar bone has been long debated. The present study reported the distribution of glycosaminoglycans (GAG) in normal and diabetic alveolar bone using histochemical techniques. Animals were rendered diabetic and killed at 2, 4, 6 and 9 weeks after injections. Tissues were stained with Alcian blue 8GX dye (pH 2.5) to demonstrate GAG and the intensity of the staining reactions compared with age-matched controls. During the experiment, weights of control animals did not change significantly; weights of diabetic animals were significantly less than initial weights from 0-6 weeks (p less than 0.001), but became nearly equal by 9 weeks. Staining intensity of diabetic bone demonstrated initial decrease (0-4 weeks) followed by a marked increase (4-9 weeks) suggesting an early decline in bone GAG levels followed by increased bone GAG levels as compared to age-matched control and initial levels. Bone GAG levels were significantly different between diabetics and age-matched controls at 2 (p less than 0.005) 4 (p less than 0.001), 6 (p less than 0.001) and 9 (p less than 0.001) weeks after streptozotocin injections. Digestion with chondroitinase AC, ABC and streptomyces hyaluronidase suggested significant differences between control and diabetic bone matrix in the levels of chondroitin 4 and 6 sulfates (p less than 0.05) and hyaluronic acid (p less than 0.001) but not dermatan sulfate. In control and diabetic bone, chondroitin sulfates were located within the bone matrix, dermatan sulfate within bone matrix and Sharpey fiber bundles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980237 TI - The fibrogenic response of adult rat lung to continuous propranolol treatment. AB - Fibrogenesis is a common pulmonary response to injury, which is usually preceded by other severe reactions, including inflammation, fluid exudation, and alveolar epithelial damage and proliferation. The purpose of this study was to examine the morphologic effects on the distal lung of a continuous propranolol treatment. Adult male rats were treated, via a subcutaneous osmotic pump, with a continuous (approximately 0.5 mg/hour) dose of propranolol HCl, a potent wide range beta adrenergic blocking agent, in saline, or saline alone. The animals were killed after one week or three weeks. Electron microscopy of the lungs of the propranolol-treated animals revealed a dramatic increase in the prominence of interstitial cells and fibers of the alveolar septa, along with focal thickening of endothelial cells and some morphologic changes in type II alveolar epithelial cells. In some animals an analysis of total protein content, as well as 3H proline incorporation into total protein and collagen was undertaken. The results of this study indicated a significant increase in total protein content and proline incorporation into collagen in the lungs of animals treated for seven days with continuous propranolol. There was no evidence of stimulated blood cells, macrophages, edema or severe epithelial damage. This study provides morphologic evidence that continuous treatment with moderate levels of propranolol results in a fibrogenic response in the peripheral lung, in the absence of typical hallmarks of severe pulmonary damage. PMID- 2980238 TI - Cytotoxic effects of 3-methylindole on alveolar epithelial cells and macrophages: with special reference to microtubular and filamentous assemblies in alveolar type I cells of bovine lung. AB - The alveolar type I cell is a major permeability barrier between the pulmonary interstitium and alveolar spaces and its thin cytoplasmic processes are greatly susceptible to injury. These cells are often observed to undergo progressive vesiculation, vacuolization and desquamation during 3-methylindole (3MI)-induced acute pulmonary edema after oral administration in goats and cattle. The present study describes proliferation of SER and the presence of polymerized tubulin in the form of microtubules arranged in large bundles shown at ultrastructural level as well as with immunofluorescence staining for tubulin in alveolar type I cells 72 hours after 3MI treatment. Such changes were not seen in pulmonary endothelial cells, alveolar type II cells, alveolar macrophages and neutrophils. The possible role of microtubules in alveolar type I cells as a mechanistic support to resist disruption against the forces of interstitial and alveolar edema is compared with alveolar type II cells, alveolar macrophages and neutrophils. The latter cells undergo dynamic movements in response to inflammatory stimuli and therefore did not show microtubules in their cytoplasm. PMID- 2980239 TI - Production and immunohistochemical characterization of monoclonal antibodies directed against renal basement membranes of rats. AB - Basement membranes were separated from rat glomeruli and purified by mild procedures, which led to a highly enriched basement membrane fraction. Here, the production and characterization of five monoclonal antibodies against tubular and glomerular basement membranes are described. These antibodies were analyzed immunohistochemically on frozen sections of rat, bovine, and human kidneys as well as on rat embryos. One monoclonal antibody (BM O II) exclusively recognized the glomerular basement membranes, another one (BM O VII) bound to tubular basement membranes and to Bowman's capsule. Three antibodies (BM O IV, BM M II, BM M III) recognized their antigens in both glomerular and tubular basement membranes as well as in mesangial cells. The BM O II antibody showed a stringent species specificity and bound only to glomerular basement membranes of the rat. The other four antibodies cross-reacted with human and bovine glomerular basement membrane and mesangial antigens; they also bound to other tissues in the developing rat embryo. Antibody binding to specific purified components of the basement membranes such as collagen type IV, laminin, heparan sulphate proteoglycan, and fibronectin was investigated by enzyme-linked immunosorbent assay (ELISA). None of these antibodies reacted with any of these known basement membrane components, indicating that the antibodies may serve as useful tools in future investigations of so far unidentified components of basement membranes. PMID- 2980240 TI - Structural and ultrastructural hepatic changes in experimental canine leishmaniasis. AB - Six 4 month-old beagles were inoculated with Leishmania donovani infantum, three of them intraperitoneally (Group A) and the other three intravenously (Group B). The animals from Group A were killed 109, 433 and 592 days after inoculation and animals from Group B 109, 171 and 334 after inoculation. The liver of each of them was examined by means of light and electron microscopy. The lesions observed in both groups were very similar, but developed more rapidly in Group B. A chronic hepatitis appeared due to infection, characterized by the presence of multiple intralobular granulomas and portal inflammatory infiltrates consisting of lymphocytes, plasmocytes and macrophages with a variable number of amastigotes. The Kupffer cells were hyperplastic and contained parasites in their cytoplasm. Gradually the hepatocytes developed a progressive cellular swelling, which during the end-stages of the process showed itself with severe nuclear degeneration, disintegration of cytoplasmic organelles, enlargement of the cytoplasmic matrix and disruption of the plasma membranes, leading to cytolysis. PMID- 2980241 TI - Changes in bone mineralization pattern: a response to local stimulus in maxilla and mandible of dogs. AB - A pilot study was carried out in order to verify the pattern of changes in mineralization of bone in the maxillas and mandibles of dogs which had a tooth extraction or luxation. Bone mineral content was determined using computerized microdensitometry. Significant changes in patterns of mineralization were found for alveolar bone, cortical bone and trabecular bone at the sites adjacent to the area of operation. These findings suggest that the three envelopes of jaw bones of the dogs are influenced by Regional activation phenomenon (RAP). These results have important implications for the design of clinical studies of periodontium. A more detailed study should elucidate the cellular mechanisms by which these changes occur. PMID- 2980242 TI - Effects of relaxin on the microvasculature of mouse mammary gland. AB - The effects of RLX on the microvasculature of the mouse mammary gland are reported. RLX (pure porcine standard NIH-RXN-P1) at a dose of 3 GPU was administered subcutaneously to virgin adult mice ovariectomized 12 days before. The mammary glands were removed 18-20 h after RLX injection and their examination by light microscopy did not reveal any substantial growth-response to the hormone. Histology and morphometry indicated striking dilation of microvessels, especially capillaries, and electron microscopy revealed an increase in the micropinocytotic vesicles, thus suggesting enhanced transendothelial transport of substances. Such phenomena, which were independent of a release of granules by mast cells, may represent an important component of the mammotrophic action of RLX. PMID- 2980243 TI - Electron microscopic observations on the formation of elastic fibres in cultures of aortic medial cells and adventitial cells. AB - The formation of elastic fibres was observed in the cultured cells derived from the tunica media and the tunica adventitia of mouse aorta. Bundles of myofilaments with dense bodies were abundantly observed in the cytoplasm of the cultured medial cells, and numerous bundles of microfibrillar components were present in the intercellular spaces. Fine granules of approximately 50 nm in diameter were observed in the bundles of microfibrillar components. It was supposed that these fine granules of elastin fused with each other and formed elastic aggregates and then formed large elastic clumps. Numerous bundles of microfibrillar components were also present in the intercellular spaces of the cultured adventitial cells. Elastic aggregates were scarcely observed in the bundles of microfibrillar components. However, large elastic clumps as observed in the medial cell culture could not be found in the adventitial cell culture. It is suggested that the formation of large elastic clumps might be related to the sheet structures or lamellae of elastic fibres in the tunica media. PMID- 2980244 TI - Effects of long-term hypothyroidism in the morphology and synaptic organization of cerebellar ectopic granule cells. AB - Abundant ectopic granule cells scattered in the cerebellar molecular layer have been observed in 30-day-old hypothyroid rats. Their morphological features indicate that they must be regarded as mature heterotopic cells arrested during their migration towards the granular layer. As their impoverished dendritic trees are identical to those seen in controls, it is unlikely that the lack of thyroid hormones played a major role in the deficient dendritic outgrowth. The study of 180-day-old hypothyroid rats revealed that although ectopic granule cells remained quite numerous, their number per unit surface was lesser than in the 30 day-old hypothyroid group. This finding may be related to the capacity displayed by heterotopic neurons to establish synaptic contacts with the components of the molecular layer. This was inferred by the presence of a peculiar synaptic cell investment formed by axosomatic and somatodendritic contacts in 180-day-old hypothyroid rats which shows that the surviving ectopic granule cells manage to adapt to an adverse milieu. PMID- 2980245 TI - Dense cored vesicles in presynaptic profiles of the rabbit dorsal lateral geniculate nucleus. AB - We are carrying out a study about the synaptic relations between identified synaptic profiles in the dorsal lateral geniculate nucleus (dLGN) of the rabbit. Here, the types of synaptic vesicle containing profiles of the dLGN are described. There are presynaptic large profiles containing round vesicles and pale mitochondria (RLP terminals) and small profiles that contain round vesicles and dark mitochondria (RSD terminals) which respectively arise from the retina and the visual cortex. Another type of presynaptic profile contains elliptical vesicles (F-boutons) which can be subdivided according to their cytoplasmic content. These F-boutons arise from dLGN interneurons. We have found different sized vesicles that have a dense core within RLP, and F terminals and a possible RSD terminal. The significance of the coexistance of pale and dense cored vesicles in the presynaptic profiles of the rabbit dLGN is discussed. PMID- 2980246 TI - Neuroendocrine lung structures and tumours: immunohistochemical study by specific markers. AB - Out of 360 lungs or lobes surgically removed, 13 non neoplastic specimens and 16 neuroendocrine (NE) tumours are investigated with immunohistochemical methods, in order to evaluate the presence of NE structures in normal and pathological human lungs. The markers used are neuron specific enolase (NSE), chromogranin (CG) and the 80 kd antigen (80 kdAg) of NE secretory granules detected by the new monoclonal Phe-5 antibody. In non-neoplastic lung specimens, clearcut immunoreactivity for all three markers appears in NE cells, neuroepithelial bodies (NEB), NE cell-hyperplasias and dysplasias. In the same specimens 4 tumourlets with analogous clearcut immunoreactivities were also observed. The NE tumours show distinct immunoreactivity for all three antisera in the 8 well differentiated cases. The 8 poorly differentiated tumours are variably immunoreactive for NSE and present low to nil staining with antisera to CG and 80 kdAg. The immunohistochemical data are interpreted according to current views about a possible relationship between NE tumours and parent normal NE lung structures. PMID- 2980247 TI - Influence of nicotine and caffeine on rat embryonic development. AB - The influence on embryonic development of nicotine and caffeine at dose levels approximating human consumption was investigated in Sprague-Dawley rats. One group of animals received nicotine administered subcutaneously by an Alzet mini osmotic pump from gestational day 6 through 12 (25 mg over 7 days; rate 149 micrograms/hr). Control animals received physiological saline in a similar manner. A second group received a single intravenous injection of caffeine (25 mg/kg) on gestational day 6. Control animals were treated with physiological saline. A further group received both nicotine and caffeine on gestational day 6 as described for the two previous groups. There were no significant differences among any of the groups with respect to maternal weight gain, litter size, embryolethality, fetal weight, or crown-rump length. The offspring of nicotine treated animals showed a significantly higher incidence of hydrocephaly when compared to the controls, but in the combined treatment group no malformed fetuses were observed. Light microscopic examination of maternal liver, kidney and placentas revealed changes in the hepatic sinusoids, glomeruli and intervillous spaces after nicotine and combined treatment. In addition, the decidua basalis was poorly developed compared to the controls. Chorionic villi and fetal kidney appeared normal in all groups. A coteratogenic effect is not evident from these findings. PMID- 2980248 TI - Leiomyosarcomas: three cases with desmin positive tumour cells, lacking ultrastructural features of smooth muscle cells. AB - A combined study of light and electron microscopy and of immunolabelling of three pleomorphic spindle cell sarcomas is presented. The light and electron microscopic features of these sarcomas were most compatible with those described for malignant fibrous histiocytoma (MFH, pleomorphic-storiform subtype). Electronmicroscopically undifferentiated and fibroblast-like cells, fibrohistiocytes and multinucleated histiocytes were observed. Characteristics belonging to smooth muscle cells were absent. By immunostaining, vimentin and desmin could be observed in tumour cells of all three cases, at least on frozen sections. Other markers such as alpha 1-antichymotrypsin, S-100 proteins, laminin, collagen IV and markers specific for skeletal muscle cells (myoglobin, actin and myosin specific for skeletal muscle) could not be demonstrated. These findings indicate that three MFH's are, in fact, poorly differentiated variants of smooth muscle tumours. It is concluded that immunophenotyping is very useful for this type of neoplasm. PMID- 2980249 TI - Abnormal development of the notochord and perinotochordal sheath in duplicitas posterior, patch and tail-short mice. AB - Interest in developmental interactions involving the notochord and perinotochordal sheath led to a comparative investigation of these structures in three mouse mutants. Alcian blue or periodic acid-Schiff staining of 9 1/2-13 days' gestational age embryos revealed a supernumerary notochordal-like mass of cells or a deflected notochord in association with duplication of the neural tube in mice of the duplicitas posterior stock. The perinotochordal sheath and basement membrane of the accessory notochordal masses were frequently defective. Patch and Tail-short embryos were also utilized for study by means of light microscopy using Alcian blue staining. In Patch embryos, although the notochord was sometimes compressed dorso-ventrally, it had an intact perinotochordal sheath and a defined, but undulated, basement membrane. Mesenchymal cells between the notochord and neural tube were occasionally replaced by cell-free space. In contrast, in Tail-short embryos a poorly formed, lightly staining or totally absent notochordal sheath was revealed. Indeed, it was sometimes difficult to distinguish the notochord from surrounding mesenchymal cells. In both the Patch and Tail-short embryos the notochord was also deflected from its medial position. In the three mutants studied, the direct or indirect effect of gene action appeared to be on the notochord and perinotochordal sheath, and the important role of these structures in abnormal axial development was established. PMID- 2980250 TI - Influence of age on the production of rat spermatozoa, on their concentration in the cauda epididymidis, and on FSH, LH and testosterone plasma levels. AB - Testis samples were taken from young (3 months), middle-aged (12 months) and aged (24 months) male rats, processed, stained and examined via a light microscope. There were no prominent abnormal germinal epithelium and interstitial tissue. However, the aging process promoted a significant decrease in the mean amount of spermatids 19 per cross tubular section, and in the amount of Sertoli cells per cross tubular section in 24-month-old rats. The concentration of spermatozoa in the cauda epididymidis showed a gradual decrease from 3 to 12 and 24 months. After hCG injection all groups of animals exhibited an increase in plasma testosterone level, although the response was smaller in 12- and 24-month animals compared to the young mature (3 months) ones. PMID- 2980251 TI - The lectin binding pattern of normal and pathologically altered synovial tissue. AB - Light-microscopical lectin-binding studies were carried out in healthy and pathologically altered synovial tissue (osteoarthrosis, rheumatoid arthritis (RA)). Seven lectins were studied: Con A, DBA, PNA, RCA, SBA, UEA-I, and WGA. Con A and WGA mark all lining cells and the majority of subintimal synovial cells. RCA and SBA stain only a portion of lining cells, regardless of the basic pathology. The lectin PNA reacts only with RA and arthrotic material, and is thus suitable for the diagnosis of inflammatory changes in synovial tissue. UEA-1 is a consistent marker for capillary endothelium and large vessels. PMID- 2980252 TI - Degenerative spondylolisthesis and degenerative scoliosis treated with a combination segmental rod-plate and transpedicular screw instrumentation system: a preliminary report. AB - Fourteen patients with degenerative spondylolisthesis and three patients with degenerative scoliosis, all of whom experienced low-back pain, lumbar radiculopathy, and/or intermittent claudication were treated with posterolateral fusion and correction of deformities using a new instrumentation system. This new spinal fixation system combines the advantages of a rod for scoliotic deformities and a plate for sagittal plane disorders. The combination rod-plates can apply multiple forces to facilitate correction of complex deformities as well as enable indirect neurologic decompression. The system allows segmental rigid fixation via transpedicular screws that is limited only to the abnormal vertebral levels to preserve the maximum number of uninvolved lumbar motion segments. The physiologic lumbar lordotic curvature is also preserved. The minimum follow-up period was 1 year. Satisfactory results were obtained in 15 patients (88%). No intraoperative complications occurred. Screw fatigue occurred in two patients at 1-year follow up examination with no sequelae. The scoliotic and spondylolisthetic deformities were reduced significantly in all patients. This method appears to assist in reducing pathologic motion and deformities that contribute to low-back pain. Compression on neural structures is relieved by thorough decompression and distraction; spinal canal anatomy is also restored. PMID- 2980253 TI - A comparative biomechanical study of spinal fixation using the combination spinal rod-plate and transpedicular screw fixation system. AB - A biomechanical study was performed comparing the stiffness and stability of the three-level combination spinal rod-plate and transpedicular screw (CSRP-TPS) fixation system with those of three anterior stabilization constructs that spanned three vertebral levels: iliac strut grafting, polymethylmethacrylate and anterior Harrington rod instrumentation (technique of Siegal et al.), and the Kaneda anterior device. The CSRP-TPS fixation system was also compared with five posterior instrumentation systems that spanned five vertebral levels: Harrington distraction rod instrumentation, segmentally wired Luque rectangular instrumentation, Cotrel-Dubousset transpedicular instrumentation. Steffee transpedicular screws and plates, and R. Roy-Camille plates under conditions of single-level instability. The relative stability of each instrumentation system was compared by mounting the fixation systems on calf spine segments containing five motion segments destabilized by complete L3 anterior corpectomies and L2-L3 and L3-L4 anterior diskectomies to simulate the two-column instability found clinically in spine fractures. Mechanical nondestructive cyclical testing in rotation, axial compression, and flexion was performed on 12 spines. All biomechanical tests were performed on a biaxial servo-controlled MTS 858 Bionix hydraulic materials testing device with a biaxial load cell. Intervertebral displacements between L2 and L4 were continuously recorded utilizing an extensometer with the knife edges placed directly adjacent to the L3 corpectomy defect during testing. This biomechanical study showed that CSRP-TPS instrumentation spanning three vertebral levels could restore the torsional, compressive, and flexural rigidity of the destabilized calf spines to that of the intact calf spines and provided more in vitro stability than either the traditional five-level Harrington distraction rod or the segmentally wired Luque rectangular instrumentation. The greatest torsional rigidity occurred with the five-level Cotrel-Dubousset instrumentation, the five-level Steffee plate and screw system, and the three-level Kaneda anterior device. In axial compression and flexural testing, the three-level CSRP-TPS system provided fixation comparable with the five-level Cotrel-Dubousset instrumentation, the five-level Steffee transpedicular screw and plate system, the five-level R. Roy-Camille plate and screw system, and the three-level Kaneda anterior device. Satisfactory levels of rigidity can be restored by three-level CSRP-TPS instrumentation under conditions of single-level instability in unstable thoracolumbar and lumbar spine fractures. PMID- 2980254 TI - In vivo creep behavior of the normal and degenerated porcine intervertebral disk: a preliminary report. AB - The viscoelastic properties of normal and surgically altered lumbar intervertebral disks from eight immature swine were examined in vivo. Rheological models were used to mathematically characterize the compressive creep-relation behavior of the disks before and after alteration by either chemonucleolysis [chymopapain (Chymodyactin) injection] or denucleation. In the normal disks, a significant modulating effect of respiration was observed that tended to increase the creep deformation response in comparison to that observed during similar in vitro and in situ studies. These results suggest that, in terms of assessing the absolute magnitude of the viscoelastic properties of lumbar disks, the influence of normal physiological function on adjacent vertebrae and surrounding tissues cannot be neglected. Preliminary results obtained from the experimentally altered disks indicated that partial denucleation primarily affected the initial stiffness behavior of the disk (24% decrease in elastic modulus), whereas chemonucleolysis caused changes in both the time-dependent (15% increase in creep rate) and instantaneous (23% decrease in elastic modulus) properties of the disk. Both partial denucleation and acute chemonucleolysis produced biomechanical changes that were comparable to grade II or slightly degenerative, age-related changes found in human lumbar intervertebral disks. PMID- 2980255 TI - Three-dimensional spinal motion measurements. Part 1: A technique for examining posture and functional spinal motion. AB - This study examines the application of a new noninvasive technology capable of accurately, reproducibly, and reliably measuring spinal motion, in real time, with 3 df. The mean values and SD of range of motion in the lumbar, thoracic, and cervical regions compares favorably with results reported by other authors. Pearson correlations yielded a statistically significant weak inverse relationship between age and range of motion in the sagittal and frontal planes (p less than .02). No correlation between sex, height, or weight and range of motion was found. There was no statistically significant evidence of organized coupling. However, a relationship between rotation and lateral motion in the thoracic region was noted. PMID- 2980257 TI - Depth of insertion of transpedicular vertebral screws into human vertebrae: effect upon screw-vertebra interface strength. AB - Improvement in the strength of the transpedicular screw-vertebra interface by increasing the depth of screw insertion may provide improved performance of spinal implants using such screws. Within human cadaveric vertebrae, we measured the failure strength of Vermont Spinal Fixator (VSF) screws under flexion or torsion loads and of Schanz screws under pull-out loads (along the screw axis). Comparisons between opposite pedicles of vertebral specimens were made at 50 vs. 80% and 80 vs. 100% of maximum available insertion depth. Mean failure strength of VSF screws at 50% depth was 75-77% (depending upon load type) of that at 80% depth; strength for screws at 100% ("to-cortex") depth was 124-154% of that at 80%. Reanalysis of the data from Lavaste shows, contrary to his conclusion, a 26% increase in strength from a 5-mm increase in screw depth of insertion. All these differences were significant (p less than 0.05) by the matched-pairs t test. Benefit from the increased strength of deeper screw placement must be balanced against possible increased operative risk. A "near-approach" x-ray view is suggested here to decrease that presumed operative risk. PMID- 2980256 TI - Three-dimensional spinal motion measurements. Part 2: A noninvasive assessment of lumbar brace immobilization of the spine. AB - The purpose of this study was to evaluate the limitation of motion as well as comfort provided by four different types of lumbar braces. The four braces were the Raney jacket, the Camp lace-up corset, a molded-polypropylene thoracolumbar sacral orthosis (TLSO), and a common elastic corset. The data revealed that all braces significantly restrict free lumbar and thoracic motion in the sagittal and frontal planes. All braces restricted lumbar motion more in the frontal than in the sagittal plane. The rigid TLSO and Raney jackets were most restrictive when compared with the Camp corset and the elastic corset. Axial rotation in the lumbar spine is normally minimal and further limitation by a brace would be negligible. All braces restricted thoracic motion despite the fact that lumbar braces were used. The elastic corset was rated the most comfortable and the Raney jacket the least comfortable. This verifies that there is an inverse relationship between a brace's ability to restrict motion and comfort. PMID- 2980258 TI - Cranial morphometry related to placement of tongs in the temporoparietal area for cervical traction. AB - The optimal placement of cervical traction tong pins to avoid cranial penetration or tong pull-off is not well established. Cranial thickness and transverse diameter in the temporoparietal area we believe to be important variables: The former was measured on 10 embalmed cadaveric specimens, the latter on 20 cleaned skeletal specimens. For each specimen, measurements were obtained at 29 points arranged within a 5 x 9-cm rectangular region above the external auditory canal. Within this region, no significant thickness variations occurred, except for one point each at the extreme posterocephalad and posterocaudad corners. Transverse diameters were constant in the area just above the pinna and gradually decreased in the anterior, posterior, and cephalad directions. No outward-flaring temporal ridge was found. We recommend pin placement just above the pinna. Anteroposterior position should probably be determined by the amount, if any, of flexion or extension desired. PMID- 2980259 TI - Dysphagia associated with cervical spine disorders: pathologic relationship? AB - Three cases of dysphagia in patients with large anterior cervical osteophytes are discussed. Although all three showed mechanical obstruction thought to be secondary to the bone growth, only one patient reported significant relief of the dysphagia after resection of the growths. The spinal etiologies of dysphagia and the operative results are discussed. PMID- 2980260 TI - Spinal subdural hematoma presenting as an epidural hematoma following gunshot wound: report of a case. AB - A man with a gunshot wound to the lumbar spine presented with leg weakness and paresthesias. Although the preoperative myelogram suggested an epidural compressive mass, a subdural hematoma was found at surgery. In addition to the case presentation, the literature was also reviewed. PMID- 2980261 TI - Continuing controversy of disc space infection. PMID- 2980262 TI - Transfusion medicine--the coming of age of a new specialty. PMID- 2980264 TI - The role of membrane glycoproteins in platelet function. PMID- 2980263 TI - The immunogenicity of platelet membrane glycoproteins. PMID- 2980265 TI - Recent developments in the Kell blood group system. PMID- 2980266 TI - The significance of IgG on the red cell surface. PMID- 2980267 TI - The significance of complement on the red cell surface. PMID- 2980269 TI - Management of patients with factor VIII inhibitors. AB - In summary, while a great deal of information has accumulated concerning the properties and natural history of F VIII inhibitors, management remains frustrating and controversial. While bleeding episodes in those who are low responders can be treated with F VIII concentrates, treatment of bleeding in high responders is often much more difficult. Current therapeutic options include F VIII concentrates of human or porcine origin in high dosage, and PCC or APCC. The choice of treatment depends on the patient's current inhibitor concentration, the type and severity of bleeding, product availability, and the preference of the medical personnel involved. However, none of the available therapeutic modalities work as well as F VIII in a hemophiliac without an inhibitor. Perhaps more promising are the immune tolerance regimens that have been developed and are now being modified and fine tuned by a number of investigators. Such regimens have reportedly eradicated F VIII inhibitors in some hemophiliacs, and have converted others from high responders to low responders, in whom bleeding episodes can be effectively treated with conventional doses of F VIII. In contrast to the F VIII inhibitors developing in hemophiliacs, those developing in nonhemophiliacs can often be eradicated with corticosteroids or immunosuppressive drugs, either alone or in combination with F VIII. Not all respond to such approaches and serious hemorrhage may still occur. Treatment of bleeding episodes has included the use of human or porcine F VIII, APCC and, in two instances, DDAVP. PMID- 2980268 TI - Factor VIII: new perspectives. PMID- 2980270 TI - By what mechanisms could prothrombin complex concentrates promote factor VIII bypassing activity in vivo? PMID- 2980271 TI - Similarities among the events occurring during the storage of red cells and platelets for transfusion. PMID- 2980273 TI - Immunohematologic problems associated with bone marrow transplantation. PMID- 2980272 TI - Platelet and red cell clearance is determined by the interaction of the IgG and complement on the cells and the activity of the reticuloendothelial system. PMID- 2980275 TI - The molecular genetics of hemophilia B. PMID- 2980274 TI - Granulocyte antigens and antibodies. AB - Granulocyte-specific antigens defined by human alloantisera are clinically important in neonatal neutropenia, autoimmune neutropenia, transfusion reactions, drug-induced immune neutropenia, and poor response to granulocyte transfusions. Many different antigens have been defined by human alloantisera using different assays. Only antigens of the N system of Lalezari are commonly studied today. The composition and location of these antigens within the granulocyte membrane are not known, but work is now proceeding on those issues. It also appears that these antigens may have some structural or functional role in the granulocyte. Although many myeloid MoAbs have been developed, there are very few reports to establish whether these identify structures that are related to the N series of granulocyte antigens. However, these MoAbs can serve as excellent reagents for better understanding the granulocyte membrane. It appears that studies during the next several years will provide exciting information about the structure-function relationships in the granulocyte membrane. PMID- 2980276 TI - Neutrophil function studies in clinical medicine. AB - A complete evaluation of neutrophil function including: chemotaxis; adhesion; aggregation; phagocytosis; granule content and degranulation; respiratory burst activity; and bacterial killing; is expensive and requires the services of a specialized laboratory. However, preliminary screening of a patient with a predisposition toward infection, can be carried out using simple and inexpensive methods. These include examination of blood films, which may prove helpful in the diagnosis of Chediak-Higashi syndrome and specific granule deficiency; the Rebuck skin window test, which estimates chemotactic defects; the NBT test, which screens for chronic granulomatous disease patients; and peroxidase staining of the blood film in order to estimate the content of myeloperoxidase, when myeloperoxidase deficiency is suspected. For final diagnosis and determination of genetic transmission and radical treatment, ie, bone marrow transplantation, specific tests are indicated. Neutrophil function studies have also proved useful in detecting diseases in which defects in neutrophil function are secondary to the primary disorder. Indeed, increased neutrophil chemotaxis has been reported in the active phase of diseases such as: familial Mediterranean fever; psoriasis vulgaris, Behcet's syndrome and Sweet's syndrome. In these disorders the neutrophil chemotaxis assay has aided in the diagnosis and follow-up, particularly in evaluating the response to antiinflammatory agents, such as colchicine. PMID- 2980277 TI - The role of the hematocrit in bleeding. AB - Low hematocrit is an often neglected cause in the pathogenesis of a prolonged bleeding time in an anemic patient. There has been ample evidence in the literature, indicating a relationship between hematocrit and the bleeding time; and that the transfusion of RBCs may correct the prolonged bleeding time often observed in anemic patients. It is unclear how a low hematocrit causes a prolongation in the bleeding time; however, two hypotheses have been put forward. First, in small blood vessels, blood flow is such that the RBCs cause the physical dispersion of platelets towards the subendothelial surface, thus promoting interaction with the blood vessel wall. Secondly, following injury to a small blood vessel, RBCs activate platelets by releasing small amounts of adenosine diphosphate (ADP) into the microvasculature following the hemolysis that often occurs during hemostasis. The fact that the hematocrit influences the bleeding time may be of clinical importance in the treatment of anemic patients, particularly those presenting a bleeding tendency. PMID- 2980278 TI - Side effects of hemapheresis. PMID- 2980279 TI - Platelet transfusion therapy. National Institutes of Health Consensus Conference. PMID- 2980280 TI - Fresh frozen plasma: indications and risks. National Institutes of Health Consensus Conference. PMID- 2980281 TI - If it changes it must be a process: study of emotion and coping during three stages of a college examination. AB - This natural experiment provides substantial evidence for the following major themes, which are based on a cognitively oriented, process-centered theory of stress and coping: First, a stressful encounter should be viewed as a dynamic, unfolding process, not as a static, unitary event. Emotion and coping (including the use of social support) were assessed at three stages of a midterm examination: the anticipation stage before the exam, the waiting stage after the exam and before grades were announced, and after grades were posted. For the group as a whole there were significant changes in emotions and coping (including the use of social support) across the three stages. Second, people experience seemingly contradictory emotions and states of mind during every stage of an encounter. In this study, for example, subjects experienced both threat emotions and challege emotions. The complexity of emotions and their cognitive appraisals reflects ambiguity regarding the multifaceted nature of the exam and its meanings, especially during the anticipation stage. Third, coping is a complex process. On the average, subjects used combinations of most of the available forms of problem-focused coping and emotion-focused coping at every stage of the exam. Different forms of coping were salient during the anticipation and waiting stages. Problem-focused coping and emphasizing the positive were more prominent during the former, and distancing more prominent during the latter. Finally, despite normatively shared emotional reactions at each stage, substantial individual differences remained. Using selected appraisal and coping variables, and taking grade point averages (GPA) into account, approximately 48% of the variances in threat and challenge emotions at the anticipation stage was explained. Controlling for variance due to the grade received, appraisal, and coping variables accounted for 28% of the variance in positive and negative emotions at the outcome stage. Including grade, 57% of the variance in positive emotions at outcome and 61% of the negative emotions at outcome were explained. PMID- 2980282 TI - Inducible expression of cytokinin biosynthesis in Agrobacterium tumefaciens by plant phenolics. AB - Nopaline strains of Agrobacterium tumefaciens contain a gene, tzs, that encodes a cytokinin biosynthetic prenyl transferase. The gene is located adjacent to the Ti plasmid virulence region and is constitutively expressed at low levels. As a result, bacteria containing tzs secrete low levels of zeatin into the medium. We find zeatin secretion to be induced more than 100-fold by acetosyringone, one of a number of naturally occurring phenolics produced by plants in response to wounding. Induction was very sensitive to the pH of the medium (optimum pH 5.5) and was due to massive overexpression of tzs-encoded cytokinin prenyl transferase activity. The relative ability of members of a set of phenols to induce tzs expression was examined and found to be parallel to that reported for activation of other virulence genes. A series of molecular cloning experiments established that virA and virG, two genes known to be essential to the virulence induction process, were necessary and sufficient for phenolic-induced tzs expression. Sequences present in the promoter region of tzs were found to be similar to those present in genes regulated by bacterial two-component positive regulatory systems. PMID- 2980283 TI - Potentiation of the contractile effects of neuropeptides in human bronchus by an enkephalinase inhibitor. AB - Many neuropeptides have been identified in human airways and these are susceptible to breakdown by endogenous enkephalinases. This study investigated the effect of enkephalinase inhibition with phosphoramidon (10(-5) M) on contractile responses to neurokinin A, eledoisin, physalaemin and substance P in human isolated bronchial smooth muscle. Contractile responses to the maximal doses of neuropeptides given were potentiated by phosphoramidon, whereas those to carbachol were unaffected. In addition, neuropeptide response curves were significantly (P less than 0.05) shifted to the left as measured by geometric mean dose ratios (95% confidence intervals) of 20.9 (5.4-81.3) n = 7 for neurokinin A; 63 (21.8-181.9) n = 6 for eledoisin, 44.7 (3.2-616.6) n = 5 for physalaemin and 6.9 (2.4-20) n = 6 for substance P. We conclude that enkephalinase inhibition in vitro significantly potentiates the contractile response to neuropeptides in human airway smooth muscle. Absence of, or decreased activity of enkephalinase could lead to enhanced effects of endogenous neuropeptides in human airways in vivo. PMID- 2980284 TI - Responsiveness of bronchial smooth muscle from asthmatic patients to relaxant and contractile agonists. AB - The mechanism underlying airway hyperresponsiveness in asthma is unknown although an abnormality in the airway smooth muscle resulting in decreased relaxation or increased contractile response has been proposed. The present study was designed to demonstrate any differences in the in vitro sensitivity of airway smooth muscle between asthmatic patients and non-asthmatic patients. Using bronchial tissue obtained by resection from mild to moderate asthmatic patients and from non-asthmatic patients, we have shown that the altered airway responsiveness seen in asthmatic patients is not reflected in airway smooth muscle sensitivity in vitro. Sensitivity of the bronchial smooth muscle to isoprenaline and aminophylline or theophylline did not differ between asthmatic patients and nonasthmatic patients, while sensitivity to carbachol and histamine was significantly reduced in tissue from asthmatic patients. These results suggest that the abnormality in asthma may not lie at the level of the airway smooth muscle. PMID- 2980285 TI - Elevated plasma concentration of neuropeptide Y and low level of circulating adrenaline in elderly asthmatics during rest and acute severe asthma. AB - Twenty-five elderly asthmatic patients attending the internal medicine emergency ward because of an acute exacerbation of asthma were sampled, prior to acute treatment, for determination of systemic venous plasma levels of noradrenaline (NA), adrenaline (A) and neuropeptide Y-like immunoreactivity (NPY-LI). Whereas NA and NPY-LI were about two-fold higher than control values, plasma A levels were not significantly increased. Twelve of the asthmatic patients were also tested at resting stable conditions and were essentially asymptomatic. All values were then similar to those of control subjects (n = 28) except for a significantly higher NPY-LI plasma level in asthmatics. In seven of these patients a near maximal physical exercise test caused significantly increased NA, A and NPY-LI plasma levels. It is concluded that the acute asthma attack is associated with elevated NA and NPY-LI plasma levels, but an impaired A response. Furthermore, that circulating NPY under these conditions has a nervous rather than adrenal origin. PMID- 2980286 TI - Cough and bronchoconstriction mediated by capsaicin-sensitive sensory neurons in the guinea-pig. AB - Neural pathways involved in cough and reflex bronchoconstriction and the effects of drugs on these airway reflexes have been studied in unanaesthetised guinea pigs exposed to aerosols of citric acid (0.13-0.78 M), capsaicin (30 microM), nicotine (9.2 mM) and histamine (0.9 mM). The number of coughs was counted during the first 3 min of exposure and the time to onset of signs of dyspnea, as an indication of bronchoconstriction, was measured. Citric acid produced bronchoconstriction and dose-dependently increased the number of coughs. Capsaicin produced both cough and bronchoconstriction. Nicotine mainly produced cough and histamine bronchoconstriction. Pretreatment of adult guinea-pigs with capsaicin (50 mg kg-1 s.c.) produced a long-lasting (greater than or equal to 10 weeks) depletion of substance P- and calcitonin gene related peptide-like immunoreactivities in the sensory nerves of the larynx, tracheobronchial tree and lung. In capsaicin-treated animals, citric acid (0.39 M) and capsaicin (30 microM) caused neither cough nor bronchoconstriction. Nicotine (9.2 mM) and mechanical stimulation still produced cough, and histamine (0.9 mM) bronchoconstriction. It is concluded that in guinea-pigs both capsaicin-sensitive (probably C-fibre endings) and capsaicin-resistant (probably rapidly adapting stretch receptors) afferent neurons may be involved in cough and reflex bronchoconstriction. PMID- 2980287 TI - The relationship between bronchial responsiveness to methacholine and bronchial responsiveness to histamine in asthmatic subjects. AB - There is widespread belief that methacholine responsiveness in asthmatic subjects is closely related to histamine responsiveness, and that the two agents may be used interchangeably in the measurement of non-specific bronchial responsiveness (NSBR). Because this view has been challenged, we have examined the repeatability of measurements of bronchial responsiveness to methacholine and histamine and the relationship between them, in groups of 20 adult asthmatic subjects. Bronchial responsiveness was expressed as the cumulative dose (in both micrograms and mmols) provoking a 20% fall in FEV1 (PD20). The predicted 95% confidence limits for the second PD20 measurement of a further pair were within the range 0.5-2.0 x first PD20 for both agents. When the two agonists were compared in the same subjects, a significant difference in potency was noted (ratio of geometric means PD20.methacholine to PD20.histamine 2.19 [micrograms], 3.43 [mmols]; p = 0.0003). Furthermore, the variance of the differences of the pairs of log PD20.methacholine and PD20.histamine measurements was found to be significantly greater than that of either the paired methacholine measurements or the paired histamine measurements (p less than 0.01). We conclude firstly that methacholine is a less potent bronchoconstrictor than histamine and secondly that, while inhalation tests with either agent broadly reflect the degree of NSBR, they measure different phenomena and cannot be used interchangeably even after allowance is made for the difference in potency. PMID- 2980288 TI - Hydroxychloroquine in steroid dependent asthma. AB - A recent case report suggested that hydroxychloroquine had a steroid sparing effect in a patient with severe chronic asthma. We have studied the effect of hydroxychloroquine in a group of nine steroid dependent adult asthmatic patients using a randomised double blind crossover comparison of hydroxychloroquine and placebo. Each patient received hydroxychloroquine (400 mg/day) or placebo for 2 month periods. The effect of hydroxychloroquine or placebo on asthma control was assessed by change in steroid dosage, visual analogue symptom scores, response to beta 2 agonist and peak expiratory flow rate (PFR) measurement. The dose of prednisolone required during hydroxychloroquine treatment did not differ from that during placebo treatment or in pre-trial period. There was no significant change in symptom scores of PFR measurement. In this study an 8 week treatment with hydroxychloroquine was of no benefit to patients with chronic steroid dependent asthma. PMID- 2980289 TI - The dissociation constant of verapamil estimated from its effect on Ca concentration-tension curves in guinea-pig tracheal muscle. AB - Concentration-tension curves for calcium ions (Ca2+) were studied in indomethacin treated guinea-pig tracheal muscle in the presence of different concentrations of carbachol in media containing 5.9 mM K+ or 40 mM K+. The effect of verapamil was investigated taking into account the steepness (the Hill coefficient) of the Ca2+ curve. When carbachol (1 microM) was added to 40 mM K+ solution, the Ca2+ concentration to produce half maximum tension (EC50) was reduced from 0.2 mM to 0.08 mM and the Hill coefficient was increased from 1.4 to 2.0, respectively. In the presence of carbachol (1 microM), the Ca2+ concentration-tension curve was not much influenced by increasing the K+ concentration from 5.9 to 40 mM K+. Verapamil (0.5 microM) shifted the Ca2+ concentration-tension curve to the right in a parallel manner under all experimental conditions, the shift being greater with curves having a smaller Hill coefficient. The dissociation constant of verapamil was not altered by carbachol when estimated from the shift of the curve if the Hill coefficient is taken into consideration. It is concluded that the relatively low susceptibility of carbachol-induced contractions to verapamil in the presence of 40 mM K+, compared with these produced by K+ alone, is not due to a decreased verapamil affinity but to improved Ca(2+)-response coupling. PMID- 2980290 TI - Corticosteroids attenuate sensitization-induced membrane changes in airway smooth muscle cells. AB - Serum from immunized guinea pigs or highly purified IgG1 induces specific changes in electrical and contractile characteristics of airway smooth muscle (ASM) cells. In this study, the effect of hydrocortisone and prednisolone pre-treatment on the events caused by a passive in vitro sensitization of ASM cells was investigated. ASM preparations isolated from male guinea pigs were exposed to immunized serum (1:10 dilution) together with different concentrations of hydrocortisone or prednisolone administered simultaneously or 30, 120 or 240 min before addition of immunized serum. During sensitization as well as during consecutive specific antigen challenges (1 mg/ml ovalbumin), both the resting membrane potential (Em) and isometric force developed by airway smooth muscle preparations were determined. Em was determined with glass microelectrodes and isometric force was simultaneously measured with a copper-berylium strain gauge. Pretreatment with both hydrocortisone and prednisolone (in the range of 10(-8) 10(-4) M) attenuated sensitization-induced transient depolarization and steady state hyperpolarization of ASM cells and decreased the sensitization-induced development of the isometric force. Furthermore, pretreatment with either steroid significantly attenuated electrical and contractile responses to a specific antigen challenge. Inhibition of the events observed during sensitization occurred at steroid concentrations as low as 10(-8) M. We have also found that the duration of exposure of ASM cells to either of the tested steroids is critical in determining the magnitude of this inhibitory effect and seems to be more important than concentration of tested steroids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980291 TI - Comparison between steady state pharmacokinetics and effects of two once-daily, slow-release theophylline formulations in nocturnal asthma. AB - The effects on bronchoconstriction, on non-specific bronchial hyperreactivity (nBH), and the pharmacokinetics at steady state of two theophylline preparations, controlled release (CR) capsules (Riker Pharmaceuticals) and UnixanR tablets (Pharmacia) were compared in patients with nocturnal asthma. Doses were individualised with the intention of achieving plasma trough concentrations greater than 8 micrograms/ml with CR-capsules. The same dose of the two formulations was taken in the morning during two 2-week periods in a randomised, double-blind, cross-over design. During a 24 h hospital study day at the end of the two periods, pulmonary function tests, blood samples, and bronchial histamine challenges were performed. Eleven patients completed the study. Doses were median 15 mg/kg (range 10-20). FEV1 and PEF were statistically significantly better (p less than 0.05) 24 h after dosing (08:00) with CR-capsules. No statistically significant changes in nBH were demonstrated. Despite no significant differences in extents of absorption, the plasma theophylline concentration fluctuations were significantly less (p less than 0.001) during treatment with CR-capsules. We found CR-capsules superior to Unixan in pharmacodynamics 24 h after dosing and in pharmacokinetics with equal bioavailability. In nocturnal asthma the pharmacodynamic differences may be eliminated with evening dosing. No statistically significant changes in nBH were observed. PMID- 2980292 TI - Adenosine-induced bronchoconstriction in the guinea-pig isolated lung: interaction with theophylline and enprofylline. AB - Effects of purine nucleosides and asthma mediators on airway tone have been examined in the guinea-pig isolated perfused lung preparation. Acetylcholine (10 pmol-0.3 nmol), histamine (1-10 nmol), adenosine (10 nmol-0.3 mumol), ATP (10 nmol-0.3 mumol) and inosine (10 mumol-0.1 mmol) all produced a dose dependent increase in lung resistance (RL) and a decrease in dynamic compliance (CDYN). ATP was equipotent with adenosine whereas inosine was about 500 times less potent. The adenosine-induced bronchoconstriction was affected neither by disodium cromoglycate (150 microM) nor by the histamine H1-receptor antagonist, mepyramine (1 microM) suggesting that histamine is not involved in this response. Furthermore, it was studied whether the xanthines theophylline and enprofylline specifically interacted with the adenosine induced bronchoconstriction. Theophylline significantly (P less than 0.01-0.001) and concentration dependently prevented both acetylcholine and adenosine-induced increase in RL. The response to 0.1 nmol acetylcholine was reduced by 32.8 +/- 8.4% (mean +/- SEM) and 58.1 +/ 4.0%, respectively, by 75 and 150 microM theophylline. Theophylline, 75 and 150 microM, also inhibited the increase in RL caused by 0.1 mumol of adenosine by 61.4 +/- 9.6% and 83.4 +/- 5.2%, respectively. Theophylline, was significantly (P less than 0.05-0.01) more potent in preventing the RL increase produced by adenosine than that by acetylcholine. Enprofylline, 30 microM, equally well as 75 microM theophylline reduced the acetylcholine-induced bronchoconstriction by 41.8 +/- 7.6% (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980293 TI - Serotonin-induced pulmonary responses in the perfused guinea pig lung: evidence for 5HT2 receptor-mediated pulmonary vascular and airway smooth muscle constriction. AB - The isolated perfused guinea pig lung was used to investigate pharmacologic characteristics of serotonin receptors in pulmonary vascular and airway smooth muscle. Serotonin caused a marked dose-related increase in pulmonary arterial pressure and peak intratracheal pressure when injected into the pulmonary artery. In contrast to serotonin, 5-carboxamidotryptamine, a 5HT1A and 5HT1B receptor agonist, and 2-methylserotonin, a 5HT3 receptor agonist produced only weak vascular and airway responses. Furthermore, vasoconstriction and bronchoconstriction elicited by serotonin were antagonized by the potent 5HT2 receptor antagonists, LY53857, ketanserin, and ritanserin. Antagonist specificity for 5HT2 receptors was demonstrated in the perfused guinea pig lung since similar responses induced by histamine were not blocked. High concentrations of serotonin were tachyphylactic on vascular but not airway constriction. Tachyphylaxis of vascular responses was not observed to an equipotent concentration of leukotriene D4 (LTD4). Thus, both the pulmonary vascular and airway constriction to serotonin were predominantly due to 5HT2 receptor activation. Furthermore, tachyphylaxis of vascular responses to serotonin might prove useful to differentiate 5HT receptor systems in pulmonary blood vessels from those in airways. PMID- 2980294 TI - Comparative distribution of cholinomimetic and parasympathetic contractile responses in the major diameter airways of dogs. AB - The distribution of cholinomimetic and parasympathetic contractile responses in the major diameter airways and the simultaneous change in lung resistance was examined in 10 mongrel dogs in vivo. The airways of animals anesthetized with chloralose-urethane were insufflated with tantalum, and responses in airway generations 0 (trachea) through 5 were assessed as the change in airway diameter (dDaw) at functional residual capacity (FRC). All animals had beta-adrenergic blockade with propranolol to block reflex sympathetic effects resulting from hypotension. Sequential dose-response curves to 10(-10) to 10(-7) mol/kg iv methacholine (MCh; n = 5) were elicited; in separate studies, stimulus-response curves were generated (1-20 Hz; n = 5) at 7 min intervals. Tantalum bronchograms were obtained at each point on the stimulus/dose-response curve for each animal. Maximal parasympathetic stimulation caused a 28.8 +/- 4.35% dDaw in trachea, a 37.2 +/- 2.33% dDaw in generation 1 airways and a 42.4 +/- 3.63% dDaw in generation 5 airways. In contrast, cholinomimetic stimulation caused substantially less narrowing of trachea (dDaw = 8.04 +/- 2.60%; P less than 0.001 vs cholinomimetic stimulation) and generation 1 airways (21.6 +/- 2.06%; P less than 0.001) and substantially less change in airway resistance (981 +/- 107% for MCh vs 1634 +/- 173% P less than 0.01) when diameter changes in generation 5 airways were comparable (45.6 +/- 7.38% vs 42.4 +/- 3.63%; P = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980295 TI - Regional differences in the reactivity of guinea-pig airways. AB - Regional differences in the sensitivity of guinea-pig airways to cholinergic and beta-adrenoceptor stimulation as well as Schultz-Dale responses were studied in vitro. Carbachol and isoproterenol produced a 27% and 39% greater response, respectively, in the trachea as compared to the bronchus. Contractions to histamine in tracheal and bronchial tissues were significantly less than those evoked by carbachol (P less than 0.05); however, lung parenchymal strips did not show any difference in maximal contractility to these agonists. Responses to antigen in airways from sensitized guinea-pigs or responses to anaphylactic mediators, LTD4 and PGD2, in normal airways did not show any differences in maximum tissue contractility (trachea and bronchus). Among the airway tissues examined, lung parenchymal strips were more responsive to these mediators as compared to tracheal and bronchial tissues. Threshold and sub-threshold concentrations of LTD4 had no effect on tracheal and bronchial airway contractions to PGD2; however, PGD2-induced contractions of the lung parenchymal strip were significantly inhibited in the presence of LTD4 (P less than 0.0001). Our data suggest that: (1) considerable regional differences exist in the reactivity of guinea-pig airways; (2) the maximum responses of bronchial airways to cholinergic and beta-adrenergic stimuli appear less than that observed in the trachea; (3) small peripheral airways appear more responsive to some mediators of anaphylaxis than are the larger central airways. Therefore, it would appear that the trachea and bronchus are more responsive to cholinergic stimuli, while the lung parenchyma is more responsive to PGD2, LTD4 and also to in vitro anaphylaxis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980297 TI - Effect of elastase instilled into the trachea on airways mechanics in guinea pigs. AB - Instilled elastase caused an inflammatory response in the lungs of guinea pigs which was observed at 6 h, 24 h, and 48 h post-treatment. The inflammation was most marked at 24 h and was characterised by a loss of epithelial cilia and detachment of epithelial cells from the basement membrane, a marked increase in polymorphonuclear leukocytes (PMNs) in blood vessels of the tracheal submucosa and an infiltration of macrophages into the parenchyma. Compared with controls, isolated tracheal preparations from 24 h and 48 h elastase pretreated animals were hyperreactive (Emax) to histamine and carbachol. This hyperreactivity persisted in tracheas from 48 h elastase pretreated animals after removal of the epithelial layer. Parenchymal strips were hyperreactive to histamine only. Tissue sensitivity (EC50) was little affected by elastase. Tracheal preparations incubated in 0.01% elastase for 3 h responded normally. In vivo responses of Raw and Cdyn to histamine were unaffected by elastase at 24 h and 48 h. However, the slope of the dose-response curve to acetylcholine was steepened 24 h after elastase instillation, but not at 48 h. In contrast to other models of inflammation elastase evokes in vitro but not in vivo hyperresponsiveness. PMID- 2980296 TI - Anaphylatoxin C3a peptide contracts human pulmonary vasculature. AB - Complement anaphylatoxin C3a C-terminus octapeptide Ala-Ala-Ala-Leu-Gly-Leu-Ala Arg (C3apep) contracted both pulmonary artery (PA) and pulmonary vein (PV) in a dose-dependent manner over the concentration range of 0.1 micrograms/ml to 100 micrograms/ml with the latter having maximal contractile activity. Removal of the terminal arginine caused complete loss of activity of C3apep. Contractions consisted of an early and a sustained component with the latter component being much greater in PV than PA. The early component was inhibited by pretreatment of tissues with the cyclooxygenase inhibitor indomethacin, or the thromboxane synthase inhibitors dazoxiben or U63,557A. The sustained contractile component was inhibited by the leukotriene antagonist FPL55712 or the 5-lipoxygenase inhibitors U60,257 (Piriprost) or nordihydroguaiiararetic (NDGA). C3apep challenge of both PA and PV resulted in the generation of leukotriene C4. These results suggest that C3apep causes contraction of human pulmonary arteries and veins by the production of thromboxane A2 and leukotrienes. PMID- 2980299 TI - National agenda for educating gifted students: statement of priorities. PMID- 2980301 TI - Pharmacokinetics of iopentol in the rat. AB - The pharmacokinetic properties and potential metabolism of iopentol have been investigated after intravenous injection in rats. The elimination was rapid, with more than 95 per cent of the injected dose recovered in urine and faeces within the first 24 h. The biologic half-life of iopentol in rat blood was 24 min. There was no significant biotransformation and only trace amounts were detected in tissues 24 hr after the injection of 200 mg I/kg or 7 days after the injection of 1 or 10 g I/kg. PMID- 2980302 TI - Metabolism and excretion of iopentol in the pig. AB - The metabolism of iopentol was examined in pigs. The search was focused on the urine and bile excreted during the first day after intravenous injection of a high dose. Chromatographic methods (both TLC and HPLC) indicated that the maximum amount of metabolites was less than 1 per cent of the given dose in urine and less than 0.3 per cent in the bile. PMID- 2980303 TI - Pharmacologic effects of iopentol after intravenous injection in healthy volunteers. Preliminary report. AB - The effects of the first injections in humans of iopentol, a new non-ionic contrast medium, are briefly reported. Detailed descriptions will be published elsewhere. Iopentol was well tolerated when injected intravenously into 24 healthy male volunteers in doses of 300 to 1,200 mg I/kg body weight and was excreted almost entirely in the urine in unchanged form. The results indicate that iopentol may be used in clinical trials in patients. PMID- 2980304 TI - Pharmacokinetics of iopentol in healthy volunteers. AB - Iopentol is a new non-ionic contrast medium. In order to study the pharmacokinetic properties, iopentol 350 mg I/ml was administered intravenously to healthy male volunteers at 3 dose levels (0.3, 0.6 and 1.2 g I/kg body weight). To evaluate renal excretion all subjects were given chromium-51 ethylenediaminetetraacetic acid (51Cr-EDTA) additionally to iopentol. The highest dose level group also received saline and 51Cr-EDTA about 3 weeks prior to iopentol. Serum, urine and faeces were sampled at different time intervals. Unmetabolized iopentol was excreted almost entirely in urine during the first 24 hours by means of glomerular filtration. A small fraction (2%) of the dose was excreted in faeces. No dose dependent pharmacokinetics was observed up to doses of 1.2 g I/kg body weight. The pharmacokinetic properties of iopentol make it a well suited contrast medium for vascular use. PMID- 2980305 TI - Synthesis, isomerism and characterization of iopentol. AB - The synthesis of iopentol is described, and the IR, NMR and FAB MS data of the compound are discussed. Iopentol exists as a mixture of several diastereomers and conformers. This topic is discussed in some detail. The kinetics of the isomerization from pure exo-iopentol to an equilibrium mixture of exo- and endo iopentol is measured by HPLC and 1H NMR spectroscopy. At 40 degrees C the endo:exo ratio is calculated from the HPLC experiments to be 0.26. The time it takes endo-iopentol to reach half of its equilibrium concentration, and the equilibration time are calculated to be 0.8 h and 5.0 h, respectively. Iopentol may exist as exo/endo pairs of four racemates. Neither GC, HPLC, IR nor high field NMR spectroscopy were suited to reveal the diastereomeric composition of the product. PMID- 2980306 TI - Determination of iopentol and other non-ionic contrast agents in biologic fluids by high-performance liquid chromatography. AB - A simple, accurate and precise high-performance liquid chromatographic method for determination of iopentol in urine and serum is presented. The modifications of the method necessary for the determination of some other non-ionic contrast agents and in other biologic fluids are also presented. PMID- 2980307 TI - Synthesis, analysis and toxicity of some compounds which may be formed during the synthesis of iopentol. AB - The process developed for the synthesis of iopentol consists of several steps. Nine possible impurities from the synthesis of iopentol have been synthesized and one has been isolated from iopentol mother liquor. These substances have been analysed by HPLC methods developed for routine control of iopentol bulk substance. Capacity factors have been calculated for each compound which relate its position in the chromatogram to iopentol. Some of the synthesized compounds have never been found in iopentol and their toxicities have therefore not been determined. Approximate intravenous LD50 values in mice have been determined for the remaining compounds. Taking into consideration these results and the small quantities in which the impurities are present, it is unlikely that they will affect the safety of the final product. PMID- 2980308 TI - Physicochemical properties and degree of protein binding of iopentol. AB - Osmolality, viscosity, partition coefficients and degree of protein binding of iopentol are presented. The properties of iopentol are compared with those of iohexol and iopamidol. PMID- 2980309 TI - Formulation and stability of iopentol. AB - The autoclaving studies which were performed to develop a formulation of iopentol are presented. The same formulation as used for iohexol was found to be suitable for iopentol. It was shown that it is essential to use a buffer such as trometamol to lower the pH during autoclaving in order to achieve a low level of deiodination after autoclaving. Results from the on-going stability studies indicate a good stability of iopentol finished product. PMID- 2980310 TI - Acute toxicity of iopentol in rodents. AB - Testing of the intravenous toxicity (LD50) of iopentol in mice and rats showed results comparable to the toxicity of non-ionic monomeric contrast media already in clinical use. Toxicity data suggest that iopentol may be a useful compound for use as an intravascular radiographic contrast medium. PMID- 2980312 TI - Experimental urography with new non-ionic dimeric and monomeric contrast agents. AB - The potential usefulness in urography of two new non-ionic dimeric ratio 6 contrast media (iodixanol and cpd. 2-5410-4A) and a new non-ionic monomeric ratio 3 contrast medium, iopentol, was investigated in a rabbit model and compared with iohexol. Urinary iodine concentrations, urine flow, estimated attenuation capacity and radiographic scoring by visual observation was determined at three dose levels in normally hydrated and at one dose level in dehydrated animals. At the high dose (500 mg I/kg) and in dehydrated animals at 200 mg I/kg the dimers yielded higher urinary iodine concentrations, an increased estimated attenuation capacity and a higher radiographic score than the monomers. Iopentol was as effective as iohexol in all the recorded parameters. PMID- 2980311 TI - Summary of the toxicologic profile of iopentol. AB - Repeat dose toxicity studies, reproductive studies and mutagenicity studies were performed to assess the safety of iopentol. It is concluded that iopentol, a non ionic contrast medium, is free from significant toxic effects. PMID- 2980313 TI - Albuminuria following nephroangiography with low osmolar contrast media in the rat. AB - The degree of albuminuria after selective nephroangiography was investigated in anaesthetized rats. Iopentol, iopromide, iodixanol and compound 2-5410-4A all caused a slight but not statistically significant increase in albuminuria when compared with the control group. After ioxaglate injection a high degree of albuminuria was observed which was statistically significant (p less than 0.05) compared with all other contrast media and control groups. PMID- 2980314 TI - Effects of iopentol, iohexol and metrizoate on the contractility of the isolated rabbit heart. AB - Fifteen isolated rabbit hearts were perfused with Krebs' solution according to the Langendorff technique. Before, during, and after perfusion of the heart with iopentol, iohexol and metrizoate at doses 1 ml, 2 ml and 4 ml (350 mg I/ml), ECG and left ventricular contractile force were registered. The contractile force was measured with a strain gauge sutured to the wall of the left ventricle. At each dose 12 perfusions were made. At each dose iohexol and iopentol produced a smaller decrease in contractile force than metrizoate (p less than 0.001). There was no difference in effect on contractile force between iohexol and iopentol. PMID- 2980315 TI - Selective injection of iopentol, iohexol and metrizoate into the left coronary artery of the dog. Induction of ventricular fibrillation and decrease of aortic pressure. AB - In twenty beagle dogs selective injections were made into the left coronary artery with iopentol, iohexol and metrizoate in doses of 4 ml, 8 ml and 16 ml. Thirty-six iopentol injections, 35 iohexol injections and 37 metrizoate injections were made. Frequencies of ventricular fibrillation were significantly lower (p less than 0.05) after iopentol (0%) and iohexol (3%) than after metrizoate (22%). Iopentol and iohexol also produced significantly less decrease in aortic blood pressure than metrizoate at the different doses. PMID- 2980316 TI - Effects of iodixanol, iopentol, iohexol and metrizoate on femoral blood flow after injection into the femoral artery of the dog. AB - The femoral blood flow was measured before, during and three min after random injection of iodixanol, iopentol, iohexol and metrizoate into the femoral arteries of 12 beagle dogs. The doses were 0.15 and 0.30 ml/kg body weight. Concentrations were 320 mg I/ml. The maximal increases in blood flow were (median values are presented and the lower dose is given first): iodixanol 9% and 32%, iopentol 28% and 66%, iohexol 43% and 77%, and metrizoate 161% and 215%. At both doses iodixanol produced significantly smaller flow increase than any other medium. At both doses iopentol and iohexol produced significantly smaller flow increase than metrizoate (p less than 0.005). There was no significant difference between iopentol and iohexol (p greater than 0.1). PMID- 2980317 TI - The influence of non-ionic contrast media on the endothelium of small arteries. A comparison of metrizamide, iohexol and iopentol. AB - In 36 rabbits, small catheters were introduced into the central artery of one ear and into the saphenous arteries of both legs. Saline, metrizamide 170 mg I/ml, iohexol 137 mg I/ml and iopentol 137 mg I/ml, all solutions isotonic with blood, were tested. Perfusion with the test-solutions was performed with 2 ml x 3 at intervals of 5 min. In the case of iopentol, a dose-response study was performed, 1 ml x 3 and 4 ml x 3 being also tested. The solutions were administered under gentle pressure and at room temperature. Blood reflow was always observed between injections. All perfusates caused endothelial damage. Minor trauma led to endothelial cell contraction. More severe trauma increased the degree and numbers of contracted endothelial cells, frequently resulting in patches of denudation, sometimes the location for thrombus formation. In the control group only few effects on the endothelium were noted. Metrizamide caused more intimal damage at all times studied than either iohexol or iopentol. The immediate effects of saline and iopentol were quite identical, but at 2 h and 24 h iopentol caused much less intimal damage than saline. PMID- 2980318 TI - Effect of four non-ionic contrast media on red blood cells in vitro. I. Morphology. AB - The influence of four non-ionic contrast media on red blood cell morphology was analyzed. The newly synthesized monomeric iopentol and the dimeric iodixanol were compared with the monomeric iopamidol and iohexol in both hypertonic and isotonic solutions at constant hematocrit of 10%. All contrast media solutions induced morphologic changes of the red blood cells and with differences between the different solutions. In isotonic solutions and with increasing concentration of contrast medium, echinocytes were produced in increasing frequency. In isotonic solution and with a high concentration of contrast medium, iopamidol, iopentol and iohexol produced 50% echinocytes and 50% stomatocytes while iodixanol produced 90-100% stomatocytes. In original solution the hypertonic iopamidol, iopentol and iohexol solutions induced desiccocyte deformation, while the nearly isotonic iodixanol still produced stomatocytes. In all the 90% volume ratio solutions (contrast medium/blood) rouleaux formation was inhibited regardless of type of red blood cell. PMID- 2980319 TI - Effect of four non-ionic contrast media on red blood cells in vitro. II. Aggregation. AB - The newly synthesized non-ionic monomeric iopentol and dimeric iodixanol were compared with the non-ionic monomers iopamidol and iohexol in both original hypertonic and isotonic solutions at a constant hematocrit of 35%. All contrast media at all volume ratios (contrast medium/blood) decreased red blood cell aggregation with no difference between the various contrast media. The degree of disaggregation of red blood cells increased with increasing concentration of contrast medium in solution. At the lowest volume ratio (2%) red blood cell aggregation was near that of normal blood. At the volume ratio 50% the disaggregatory effects following both isotonic and hypertonic solutions were pronounced. At a high molar concentration both the isotonic iodixanol and the hypertonic solutions of iohexol, iopentol and iopamidol induced an almost total inhibition of red blood cell aggregation. This indicates that the concentration of contrast medium (chemotoxic effect) for non-ionic contrast media is more important for the disaggregatory effect than the osmolality. PMID- 2980321 TI - Contrast media research. An investment for the future. PMID- 2980320 TI - Effect of four non-ionic contrast media on red blood cells in vitro. III. Deformability. AB - The influence of four non-ionic contrast media and one ionic contrast medium was analyzed on red blood cell deformability in vitro. The newly synthesized non ionic monomeric iopentol and dimeric iodixanol were compared with the currently used non-ionic monomers iopamidol and iohexol and the ionic monomeric diatrizoate at a constant hematocrit of 8%. The effect on red blood cell deformability was analyzed in 50% volume ratio solutions of both original hypertonic and isotonic contrast media solutions. The samples were studied in a red blood cell filtrometer through new Mynipore microsieves using an improved procedure of data processing. Diatrizoate induced significantly more red blood cell rigidification compared with all non-ionic contrast media solutions and normal blood. There was no difference between the investigated non-ionic contrast media, nor between the non-ionic contrast media and normal blood without additive. PMID- 2980322 TI - Effect of iopentol on coagulation and platelet function in vitro and in vivo. AB - The effect on hemostatic parameters of two non-ionic contrast media, iopentol and iohexol in concentrations of 350 mg I/ml, were tested in vitro. The new medium iopentol had similar effects to those of iohexol, both with respect to the intrinsic coagulation system and platelet aggregation. As part of a human pharmacologic study the effects of iopentol on hemostatic parameters were tested in 8 volunteers in vivo. Iopentol induced a slightly decreased coagulability and aggregability in venous blood. It was concluded that the effects were of very little consequence for hemostasis as a whole. PMID- 2980323 TI - Effect of iopentol on acetylcholinesterase activity and thrombin time. Comparative studies. AB - Iopentol is a new non-ionic monomeric intravascular radiologic contrast medium which is undergoing clinical trials. Studies on other iodinated contrast media of various chemical structures, both closely similar to and widely different from iopentol, have shown that some in vitro tests of chemotoxicity correlate very well with clinical systemic toxicity. Some of these tests have been carried out on iopentol and are reported here. Comparison has been made in each case both with a conventional ionic contrast medium, sodium iothalamate, and with another well established medium of low toxicity already in use, iohexol. The results indicate that iopentol will have low clinical toxicity. PMID- 2980324 TI - Dispersion versus absorption (DISPA) method for automatic phasing of Fourier transform ion cyclotron resonance mass spectra. PMID- 2980325 TI - Effect of time-domain dynamic range on stored waveform excitation for Fourier transform ion cyclotron resonance mass spectrometry. PMID- 2980326 TI - Fast atom bombardment mass spectrometry of polar metabolites of organophosphorus pesticides. PMID- 2980327 TI - Translational energy release and stereochemistry of steroids. X. The effect of the 5,6-double bond on the elimination of angular methyls and water from metastable molecular ions of epimeric steroid alcohols. PMID- 2980328 TI - Analysis of nonderivatised peptides by thermospray using a magnetic sector mass spectrometer. PMID- 2980329 TI - Analyses of carcinogen-modified oligonucleotides by fast atom bombardment/tandem mass spectrometry. PMID- 2980330 TI - Trace level measurement of enkephalin peptides at the attomole/femtomole level by FAB-MS. Optimization of FAB matrix conditions. PMID- 2980331 TI - The analysis of small proteins in the molecular weight range 10-24 kDa by magnetic sector mass spectrometry. PMID- 2980332 TI - The structural elucidation of bacterial capsular polysaccharides using laser desorption ionisation Fourier transform ion cyclotron resonance spectroscopy. PMID- 2980333 TI - Thermospray mass spectrometry of aldicarb and its metabolites. PMID- 2980334 TI - Temperature dependence of neburon mass spectra using thermospray or a moving-belt interface. PMID- 2980336 TI - Diagnostic daughter ions for steroidal 4-en-3-ones. PMID- 2980335 TI - Assessment of the effect of growth temperature on the lipid composition of Serratia marcescens using laser desorption mass spectrometry. PMID- 2980337 TI - Translational energy release and stereochemistry of steroids. XII. The differences in the origin of m/z 124 and 163 key ions in epimeric 11-hydroxy steroids of the delta 4-3-keto series. PMID- 2980338 TI - Mass spectral evidence for the electrophilicity of N-ethylmaleimide and thiobenzoyl groups. PMID- 2980339 TI - Reproductive hazards of the workplace. PMID- 2980340 TI - DBCP: eleven years later. PMID- 2980341 TI - Monitoring of ovulation in the assessment of reproductive hazards in the workplace. AB - If women are exposed to reproductive hazards in the workplace, some disturbances of ovulation are expected. Assessment of ovulation requires monitoring to distinguish normal and abnormal ovarian cycles. Menstrual interval and basal body temperature charts are inadequate for identifying many abnormal cycles. A late luteal endometrial biopsy or a single mid-luteal morning progesterone level each appears to be only 80% accurate in distinguishing normal and abnormal cycles. The complete "cycle profile" of daily ovarian ultrasonic scans and daily serum levels of reproductive hormones appears to be a definitive approach to characterizing ovarian cycles, but this demanding regimen would not be applicable to monitoring large populations in the workplace. An ovarian cycle monitoring protocol consisting of daily measurement of salivary or vaginal electrical resistance, mid cycle urine testing for the luteinizing hormone surge, and daily luteal phase salivary progesterone levels would provide a practical comprehensive corroborative assessment of ovarian cycles in such populations of women. This monitoring of ovarian cycles would aid in the early detection of reproductive hazards and medical conditions that might present as an ovulatory disturbance. PMID- 2980342 TI - Assessment of menstrual variability in working populations. AB - Though the relationship between exposure to chemicals and damage to ovarian tissue has been demonstrated, there have been few attempts to assess the effect of occupational exposures on menstrual cycles. Previous research has reported the mean cycle lengths and duration of flow for large populations. However, additional research is needed to analyze the variability of cycles of individual working women. Cycle length is the most assessable indicator of ovarian damage and subsequent effects on menstrual patterns, but the reliability of self-reports of cycle length has been shown to be poor. Improved methods to accurately assess individual menstrual variability are needed. Educational programs should be provided for workers to increase their knowledge of the effects of chemicals on reproductive functioning and ultimately to increase their sensitivity to recording accurate data on menstrual patterns. Numerous factors can affect the hypothalamic-pituitary-gonadal balance. The simultaneous effects of these factors and exposures to chemicals in the workplace need further investigation. PMID- 2980343 TI - Sperm evaluation in human genetic monitoring. AB - Health hazards, including reproductive hazards, resulting from exposure to mutagenic chemical or physical agents are a major concern. Human genetic monitoring is a relatively new approach to the evaluation of human population exposure to mutagens that is based on the use of short-term tests with human cells (including sperm), tissues, and body fluids for genetic damage. Evaluation of sperm for count, percentage of morphologically abnormal forms, and frequency of heads containing zero, one, or two fluorescent bodies have been incorporated in genetic monitoring studies. In this paper, the application of human genetic monitoring to the evaluation of germ cell mutagen exposure is discussed. The purposes and limitations of human genetic monitoring studies are presented. Some of the practical issues involved in the use of sperm studies in genetic monitoring are described through the examples of two studies in different settings that were conducted by the author's laboratory. The first was a study of Autopsy Service workers within the resident institution of the laboratory. The second was a study of workers exposed to coal tar pitch volatiles at an industrial plant approximately 1,000 miles from the laboratory. PMID- 2980344 TI - Longitudinal study of semen quality of unexposed workers. I. Study overview. AB - A longitudinal study of 45 men was conducted evaluating the semen quality of monthly samples collected over 9 months. The statistical variation of sperm count, semen volume, percentage of motile sperm, sperm velocity, sperm morphology, and sperm viability, assessed by both the vital stain and the hypoosmotic swelling (HOS) assay, were each evaluated using intraclass correlations and coefficients of variation. Sperm count and semen volume had large intraclass correlations (62% and 60%, respectively), indicating that if a subject has a high count or volume he will tend to continue to have high counts or volumes. On the other hand, sperm velocity had an intraclass correlation of only 16% indicating that fluctuations within a subject were nearly as large as fluctuations from subject to subject. The remaining parameters had intraclass correlations ranging from 42% to 47%. Sperm count, percent motile sperm, and semen volume each had large coefficients of variation (both between and within subjects). These variables, especially count, had relatively poor precision. Sperm velocity, percent motile sperm, percent normal morphology, the HOS assay, and the vital stain assay had lower coefficients of variation, indicating greater precision. PMID- 2980345 TI - The effects of ethylene dibromide on semen quality: a comparison of short-term and chronic exposure. AB - Two occupational field studies were conducted to determine the effects of ethylene dibromide (EDB) exposure on male reproductive potential. The first study was a longitudinal study of 10 EDB-exposed forestry employees and 6 unexposed men conducted in Colorado in the summer of 1983. The exposure time was approximately 6 weeks. The second study was a cross-sectional study of 46 EDB-exposed papaya workers and 43 unexposed men conducted in Hawaii in December 1983 in which the average term of employment was about 5 years. In the longitudinal study, sperm velocity decreased in all 10 exposed men and in only two unexposed men. Semen volume was also decreased in 9 of the 10 exposed men (there was no change in the other man); only two unexposed men had a decrease in their semen volume. The longer term EDB exposure resulted in decreases in sperm motility and viability, suggesting that the short term exposure may slow sperm velocity, but longer exposures cause immotility and cell death. An apparent decrease in semen volume that was observed in the longitudinal study was not statistically significant in the cross sectional study of workers having chronic exposure. However, a significantly higher semen pH was observed in the exposed men compared to the unexposed men in the cross-sectional study. The results from both studies suggest that the accessory sex glands may be affected by EDB exposure. PMID- 2980346 TI - Measuring human chorionic gonadotropin for detection of early pregnancy loss. AB - Elucidation of the primary molecular structure of hCG, coupled with monoclonal antibody technology, has permitted the construction of a partial map of hCG surfaces. Based on this information, two-site immunoradiometric assays have been developed which permit the measurement of intact hCG and its subunit molecular forms with unprecedented sensitivity and specificity. These assays have been employed in a determination of the incidence of early pregnancy loss in a normal population with the finding that 22% of all conceptions producing measurable hCG terminate before becoming clinically evident. PMID- 2980347 TI - Using time-to-pregnancy data to study occupational exposures: methodology. AB - Occupational and environmental influences on fertility are for the most part unstudied, partly because sensitive methods for studying them have not been developed. We are developing a measure of fecundability, the monthly probability of pregnancy, by studying time to pregnancy, the number of noncontracepting menstrual cycles each couple requires to conceive. The relationship of this measure of reproductive impairment with others, such as spontaneous abortion, is not known. Preliminary data from two sources suggest that reduced fertility is not highly correlated with increased risk of spontaneous abortion, despite predictions to the contrary from the toxicology literature. A current study of occupational exposures of dental assistants will address questions of data quality by providing comparisons of brief responses from mail questionnaires and telephone interviews with very detailed data from telephone interviews. This study also will allow estimation of the magnitude of two potential selection biases: selection of only planned pregnancies (time-to-pregnancy data for accidental pregnancies are not meaningful), and selection against highly infertile and sterile couples (when studying currently or previously pregnant women, sterile couples are not represented at all and highly infertile couples are underrepresented). PMID- 2980348 TI - Semen quality in workers producing reinforced plastic. AB - A comparison of semen parameters from 25 workers at a reinforced plastic production plant and from 46 age-matched male fertility patients was carried out. For a period of more than three months before the semen analyses, the 25 workers had been exposed to styrene and acetone in concentrations exceeding the Danish threshold limit values. Serum concentrations of follicle stimulating and luteinizing hormones, semen sample volume, and concentration of sperm showed no differences between the two groups. Significant differences between the two groups were found in the percentage of live sperm, immotile sperm, and normal sperm. The reinforced plastic plant workers showed, in terms of semen quality, better values in the percentages of live sperm and immotile sperm, whereas they had a reduced percentage of normally shaped sperm. It was found that the reinforced-plastic workers had increased percentages of the amorphous and the pyriform sperm head shapes as compared to the fertility clientele. In sperm midpiece and sperm tail defects, the two groups showed similar values. From the data it could be concluded that work with production of reinforced plastic might result in an increase in the number of abnormal sperm heads. PMID- 2980349 TI - Stainless steel welding and semen quality. AB - Questionnaire studies of patients from fertility clinics suggest that welders may have an increased risk of reduced semen quality. In this study, welders and nonwelders from the same plants were asked to provide blood, urine, and semen samples. Urine was analyzed for chromium and nickel, and for mutagenic activity and metal concentration; blood for metal concentrations, immunoglobulin G, total protein, and measures of genotoxicity in lymphocytes; and semen was evaluated by standard semen analysis. Results of the semen evaluation, presented here, showed no difference in semen quality between welders and nonwelders. Because the metal dust exposure of nonwelders in the plant may be higher than that in the general population, welders were also compared to referents not working in the metal industry. Again, no decrease in semen quality associated with welding was demonstrated. PMID- 2980350 TI - Endocrine evaluation in the assessment of male reproductive hazards. AB - Male reproductive function requires the integrated functioning of the hypothalamus, pituitary, and testis. The disturbance of endocrine function at any of these levels may result in hypogonadism and infertility. The clinical and laboratory evaluation of these disorders is reviewed here. PMID- 2980351 TI - Monitoring fertility to detect toxicity to the male reproductive system. AB - This paper describes measures of fertility for detecting toxicity to the male reproductive system. The limitations of fertility monitoring are noted as well as the advantages and disadvantages of various fertility measures. Time to pregnancy and standardized fertility ratios, two fertility measures about which more is known, are discussed in greater depth. PMID- 2980352 TI - Lessons from a surveillance program of semen quality. AB - The discovery in 1977 that dibromochloropropane (DBCP) could induce male sterility led to an interest in the surveillance of semen quality in men working in a pesticide manufacturing plant. The program consisted of an initial questionnaire, physical examination, and serial semen analyses. The success of the surveillance was limited by economic considerations and protocol compliance problems. Based on the experience with this program, recommendations for future surveillance efforts are presented. PMID- 2980353 TI - Use of the SPA in assessing toxic effects on male fertilizing potential. AB - The sperm penetration assay (SPA), which involves the use of zona-free hamster eggs to assess fertilizing potential of human spermatozoa, has the capability of evaluating the functional capacity of sperm. If men are exposed to a toxic insult that could result in impaired fertility, this has been monitored historically by determining the routine semen parameters of concentration, motility, and morphology. The lack of availability of the SPA for evaluating toxic exposure in field studies has hampered its utilization. The purpose of this article is to illustrate the use of the SPA in a specific field study and to describe two recent innovations in the SPA that make its use more accessible for field studies. These two innovations are shipping semen samples in TEST-yolk buffer by overnight express to a central laboratory for testing and using frozen hamster eggs in the test. The report is divided into the following three major sections: I. Kunia study; II. TEST-yolk buffer modifications of SPA; and III. Use of frozen eggs in SPA. PMID- 2980354 TI - Assessing female fertility. AB - A concern exists that infertility is on the rise among American couples. While secular trends do not support this, there is reason to believe that deferral of child bearing is associated with a delay in conception. This paper reviews the data on factors that interfere with fertility and fecundity, such as maternal age and occupational exposures. Also discussed are epidemiological tools and strategies for evaluating the various components of female fertility and for identifying adverse influences. PMID- 2980355 TI - NIOSH epidemiologic studies of pregnancy outcomes. AB - The conduct of pregnancy outcome studies among occupationally exposed populations is a relatively new research area with many methodologic and pragmatic difficulties. This paper reviews the experience of conducting reproductive epidemiologic research at the National Institute for Occupational Safety and Health (NIOSH). The advantages and disadvantages of the three epidemiologic study types most often conducted at NIOSH are discussed: 1) surveillance studies, 2) descriptive studies, and 3) industrywide studies. Some of the problems inherent in the conduct of industrywide investigations are illustrated in a discussion of a reproductive outcome study of video display operators currently underway. These problems include sample size, referent availability, exposure and outcome validation, tracing, analytic methods, and cost. PMID- 2980356 TI - Computer modeling of human fertility: the impact of reproductive heterogeneity on measures of fertility. AB - As a result of the paucity of biological markers for both reproductive events and exposure to reproductive toxicants, it is likely that individual members of populations will be misclassified with respect to reproductive performance and xenobiotic exposure. A four-parameter computerized model of fertility (frequency of intercourse, male fecundity, female fecundity, and spontaneous abortion) was developed to explore the effect of misclassification of populations on several measures of fertility. The measures of fertility explored include cumulative percent pregnant, time to pregnancy, cycle specific fertility, and fecundability ratio. The cumulative percent pregnant and time to pregnancy appeared to be insensitive to small changes in reproductive competence. The fecundability ratio decreased in proportion to the size of population exposed to a reproductive toxicant. The three reproductive parameters, time to pregnancy, cumulative percent pregnant, and fecundability, do not appear useful as measures of reproductive heterogeneity in populations. The reproductive heterogeneity of a population was best defined by the change in cycle-specific fertility rate over the period of observation. These simulations suggest that the change in the cycle specific fertility rate should be evaluated and comparable in all population groups over the period of observation to assure reproductive homogeneity. PMID- 2980357 TI - The National Toxicology Program chemical nomination selection and testing process. AB - The NTP is an interagency program of the Federal Government which coordinates toxicological programs at the NIH (NIEHS), FDA (NCTR), and CDC (NIOSH) with input from NCI, NIH, OSHA, CPSC, EPA, and ATSDR. The NTP has the capability to completely characterize the toxicologic profile of a chemical, including studies of chemical disposition, genetic toxicity, immunotoxicity, teratology, reproductive toxicity, carcinogenicity, neurotoxicity, and specific organ toxicity. The NTP encourages nominations of chemicals of human health concern from all sectors of the public, including industry, labor, and the general public. The specific process of nomination, evaluation, and selection of chemicals for testing by the NTP is described. It is a multicomponent system with several evaluations and a public peer review step to assure adequate consideration of all nominated chemicals. The results of NTP studies are all peer reviewed and available to the general public as well as to the scientific community. PMID- 2980358 TI - NIOSH selection of chemicals and study publications: setting priorities for reproductive research. AB - The National Institute for Occupational Safety and Health (NIOSH) prioritizes topics for reproductive research in order to allocate limited resources to the most important issues. In the past, we have relied on an informal network including our own project officers, toxicologists, and the public to nominate chemicals to study. Setting priorities has consisted of the evaluation of research ideas by criteria such as the chemical's probable toxicity and the extent of exposure to the chemical in the workplace. NIOSH researchers are developing a system for evaluating large databases of toxicologic and occupational exposure information to identify potential reproductive toxins. When this system is operational it will provide a systematic way to nominate chemicals for study. PMID- 2980359 TI - Food and Drug Administration guidelines for reproductive toxicity testing. AB - The Food and Drug Administration generally requires reproductive toxicity testing of all new drugs to be used by pregnant women or women or men of reproductive potential. These requirements may vary among the centers within the FDA. Reproductive and developmental toxicity is usually tested in one or two animal species and is divided into three segments to represent treatment throughout the reproductive process. The FDA monitors adverse drug effects on human reproduction through postmarketing surveillance. PMID- 2980360 TI - Reproductive effects of chlorpromazine exposure to female mice: cell proliferation disadvantage revealed by the Chimera Embryo Assay. AB - Administration of chlorpromazine-HCl at 5 to 15 mg/kg bodyweight to pregnant CD-1 mice at 24 h after human chorionic gonadotropin (hCG) (20-23 h after mating) inhibited blastocyst formation and reduced the cell number of embryos recovered at 95 h after hCG. When embryos are recovered at the two- to four-cell stage (48 50 h after hCG) and cultured for an additional 47 h (to 95 h after hCG) or 72 h (to 120 h after hCG), blastocyst formation and embryo cell number were similarly reduced. When the dose range was reduced to 0.5 to 2 mg/kg bodyweight, no significant effect of the drug was observed on blastocyst formation or on embryo cell number. However, when aggregation chimeras were formed between embryos recovered from drug-exposed females and from untreated females, a decrease in cell proliferation rate of the embryo from the drug-exposed female was observed at a dose of 2 mg/kg bodyweight. This result indicates that exposing pregnant mice to chlorpromazine-HCl at doses as low as 2 mg/kg bodyweight can induce a potential for decreased cleavage rate in their pre-implantation embryos that can be revealed by challenging those embryos by direct contact with embryos from nonexposed females. Finally, when four-cell stage embryos recovered from untreated females cultured in the presence of chlorpromazine (0.1-25 mM), blastocyst formation and embryo cell number were significantly reduced in a dose dependent manner. This last result suggests that in vivo the drug may act directly on the embryo from the pronuclear stage to the early morula stage of development. PMID- 2980361 TI - Role of testicular versus epididymal toxicity in the induction of cytotoxic damage in Fischer-344 rat sperm by methyl chloride. AB - Exposure of male Fischer-344 (F-344) rats to methyl chloride (MeCl) results in testicular and epididymal toxicity and the induction of both pre- and postimplantation embryonic loss; the preimplantation loss is caused by cytotoxic damage to sperm that leads to failure of fertilization (Toxicol Appl Pharmacol 1986; 86:124-130). The present study examined whether the cytotoxicity of MeCl to sperm is due to the testicular or epididymal toxicity of MeCl. Groups of 18 males were exposed to 3000 ppm MeCl 6 h/day for 5 days, with and without concurrent treatment with the anti-inflammatory agent 3-amino-1-[m-(trifluoromethyl)phenyl] 2-pyrazoline (BW755C; 10 mg/kg, i.p. 1 h pre- and postexposure); BW755C was used to inhibit the epididymal toxicity of MeCl. Control groups were untreated or injected as described above with BW755C. Six males from each group were killed weekly for 3 weeks. Toxic effects of MeCl on the testis were demonstrated by decreased relative organ weight (week 3), testicular histopathology (weeks 1-3) and decreased daily sperm production (weeks 1-3); these effects were not prevented by BW755C. In both the MeCl and the MeCl + BW755C treatment groups, tubules devoid of sperm were observed in regions 4 and 5 of the epididymis at week 2, and in regions 6A and 6B at week 3. Sperm were present in the vas deferens of both groups at week 3 in decreased numbers and had decreased motility and more frequent morphologic abnormalities compared to untreated controls. In conjunction with known epididymal transit times for F-344 rat sperm, these data indicate that the induction of preimplantation loss by MeCl at weeks 2 and 3 postexposure is likely to result from cytotoxic effects on sperm located in the testes at the time of exposure. PMID- 2980362 TI - Ionizing radiation and human reproduction. PMID- 2980363 TI - Effect of oocyte number and rate of atresia on the age of menopause. AB - Menopause occurs when oocyte number falls below the threshold required for ovarian function. The age of menopause is decreased by some drugs, some occupational exposures, and cigarette smoke (both active and passive). Using data collected by Block (Acta Anat (Basel) 1954; 14:208), we have constructed four, two parameter models to explore the effect of oocyte number and rate of atresia on age at menopause. The models used are: linear [O(A) = OB + R * A]; natural log [O(A) = R * In (A)]; exponential [O(A) = OB * exp(R * A)], and power [O(A) = OB * Age(R)]. OB = oocyte number at birth, ATR = rate of atresia, and O(A) = number of oocytes at age A. The parameters for the four models are: (formula; see text) Each model has different behavior with respect to alterations in oocyte number and rate of atresia on age at menopause. Animal studies suggest that ovarian failure is only weakly dependent on oocyte number, consistent with the power and exponential models. These suggest that the decrease in age at menopause following xenobiotic exposures results from an increase in the rate of atresia. PMID- 2980364 TI - Female reproductive risk assessment. PMID- 2980365 TI - Possible animal models of follicular development relevant to reproductive toxicology. AB - Models of follicular development in rodents that may be applicable to reproductive toxicology are considered. The value of evaluating changes in follicular numbers during the estrous cycle is stressed. Most of the methods involve in vivo manipulations. However, the use of enzymes to dissociate intact follicles from the ovary and their subsequent in vitro development in the presence or absence of xenobiotics offers an alternative, attractive approach. PMID- 2980366 TI - Corpus luteum: animal models of possible relevance to reproductive toxicology. AB - The presence of a normally functioning corpus luteum is an essential requirement for the maintenance of gestation in mammals. The chief function of the corpus luteum in all species is to synthesize the steroid hormone progesterone that is necessary for implantation and for the subsequent development of the fetoplacental unit. Activation and maintenance of luteal function involve pituitary, placental, and ovarian hormones. Perturbation in the secretion and/or action of any of these luteotropins by exogenous compounds can profoundly affect the steroidogenic capacity of the corpus luteum. Abnormal luteal function causes failure of implantation and embryonic wastage. Both in vitro and in vivo models for assessing luteal function are available; while the former are more convenient, the model of choice for toxicological studies is one in which in vitro findings can be easily confirmed in whole animals in vivo. This article is concerned primarily with the techniques used in basic research that might be useful for the evaluation of luteal function in reproductive toxicology. PMID- 2980367 TI - Histological assessment of follicular development and its applicability to risk assessment. AB - The objective of this article is to describe histological methods currently used to study ovarian physiology that may have applicability to reproductive toxicology. Histologic techniques are, for the most part, inexpensive and easy to perform. Many of these techniques require little equipment and can be readily implemented in small laboratories. The resulting histological preparations are permanent, can be used over again for several different types of analyses, and provide objective, quantifiable endpoints. These techniques have been extremely useful for studying the ovary. PMID- 2980368 TI - A method to obtain maternal-fetal plasma samples using a microsampling technique in the rat: transplacental passage of cefoxitin. AB - A microsampling technique that allows taking blood samples from the umbilical vein of the pregnant rat is described. Such techniques are needed in order to allow pharmacokinetic and embryo exposure to be correlated with teratogenic endpoints. Cefoxitin was administered intravenously (300 mg/kg) into tracheotomized, pentobarbital anesthetized dams on day 21 in gestation. Blood samples were collected via the carotid artery from the dam and the umbilical vein of the fetus at designated times. Up to three samples of 20 to 30 microliters each, were taken from individual fetuses at 20-min intervals. With few exceptions, fetal cefoxitin concentrations were homogeneous at each sampling period. Fetal concentrations were low compared to maternal concentrations as seen by the small fetal/maternal area under the curve ratio (0.053 +/- 0.006). PMID- 2980370 TI - Lack of syringe toxicity to amniotic fluid cells in culture. AB - Toxicity to amniotic fluid from many brands of syringes has been reported in the past few years. Thirty amniocentesis samples were exposed to Becton-Dickinson syringes for varying lengths of time and evaluated for growth rate. Six amniocentesis samples were exposed to the syringes for prolonged periods at varying temperatures and also evaluated for growth rate. No evidence of syringe toxicity was found for exposure periods up to 24 h, regardless of storage temperature. PMID- 2980369 TI - Congenital malformations and intrauterine growth retardation in streptozotocin induced diabetes during gestation in the rat. AB - Diabetes was induced in Wistar rats by single intraperitoneal injections of streptozotocin (STZ) (50 mg/kg, in citrate buffer pH 4.7) at both preimplantation and organogenetic stages. The controls were either buffer treated or, following STZ administration, injected intraperitoneally with 2 to 6 IU of insulin daily until term. Fetuses collected on day 20 were found to have several malformations. Major abnormalities included gastroschisis and evisceration, maxillary hypoplasia and interatrial, and interventricular septal defects. Intrauterine growth retardation was present in most of the diabetic groups. Heavier placentae and shorter umbilical cords were also observed. Malformations and intrauterine growth retardation were numerous and severe in the preimplantation treatment groups. Insulin administration significantly improved the fetal body weight but did not change the malformation rate in the group made diabetic during organogenesis. It appears that the duration of hyperglycemic state plays a crucial role in malformation and growth retardation, and meticulous control of blood sugar from early stages has significant beneficial effects on fetal development. PMID- 2980371 TI - Neuroendocrine mechanisms controlling the onset of female puberty. AB - This article describes the major neuroendocrine developments that are associated with the normal onset of puberty in the female rats. Although differences exist between rats and humans with regard to development and function of the reproductive system, there are major basic similarities. This makes the rat a valuable laboratory model in which to evaluate the effects of potentially hazardous substances on normal sexual maturation. A systematic experimental approach is presented to show how the study of reproductive development in the female rat can aid our understanding of the processes by which these substances could disturb normal puberty in the human female. PMID- 2980372 TI - Neuroendocrinology of female reproduction: review, models, and potential approaches for risk assessment. AB - The central role of the mammalian hypothalamic-pituitary axis in regulating female reproductive cycles is reviewed. A variety of animal models and techniques that offer increased sensitivity, speed, and flexibility over traditional reproductive toxicologic approaches for short term testing or screening are discussed, including the pivotal analysis of gonadotrophin releasing hormone (GnRH) pulse generator activity in vivo and in vitro. Other neuroendocrine techniques that require further development, but provide potential approaches to demonstrate specific sites or mechanisms of action for toxic effects on the hypothalamic-pituitary axis are suggested. PMID- 2980373 TI - Critical evaluation of methods for detection and assessment of estrogenic compounds in mammals: strengths and limitations for application to risk assessment. AB - Methods for assessing proestrogenic and estrogenic activity of xenobiotics (environmental pollutants and therapeutic agents) are critically evaluated. The uterotropic assay in vivo and the binding affinity to the uterine estrogen receptor in vitro are still the preferred methods by most investigators. Because metabolism could markedly alter the activity of estrogens and proestrogens, in vitro methods that do not consider such possibilities have only limited utility. An in vitro method, which attempts to satisfy that requirement by combining metabolic capability with binding to the estrogen receptor, has become available. However this method still needs considerable refinement and the assessment of its potential and limitations. In addition, there is a need for a facile method for determining whether a compound is a frank estrogen or a partial agonist/antiestrogen. These methods are considered in respect to their application for risk assessment. PMID- 2980374 TI - Update on caffeine. PMID- 2980375 TI - Nitrous oxide and male fertility. PMID- 2980376 TI - Mutagenesis in murine spermatogonia by MOPP therapy. AB - Male mice were given mechlorethamine (2.0 mg/kg) followed immediately or 1, 2, 4, or 8 h later by procarbazine (100 mg/kg) and vincristine (0.67 mg/kg) in a protocol that stimulated the drugs and dosages used in a single cycle of clinical MOPP therapy. Seven weeks later and continuing for the next five weeks, the mice were mated to females and the dominant lethal mutant frequencies determined in their offspring by an in vitro assay. Significant (Student's t-test, p less than 0.05) mutant frequencies were detected when the interval between drug administration was between 2 and 8 h. In another series of experiments, mice were injected with these drugs using a 4-h interval between mechlorethamine and the vincristine and procarbazine (MOPP), two cycles of MOPP separated by one week, one cycle of MOPP and 2.0 Gy x-radiation separated by one week, or 2.0 Gy x radiation alone. When each group was assayed for its dominant lethal mutant frequency, all were found to have similar, highly significant (0.001 less than p less than 0.005) induction. Together these data suggest that stable mutagenic lesions are induced in spermatagonial stem cells by both x-radiation and chemotherapeutic drugs and the frequency with which they are produced can be approximated by a single treatment. Clinical strategies to minimize genetic effects should concentrate on the sequence and timing of drug administration on those days when the drugs are administered in combination. PMID- 2980377 TI - Looking at trees until you see the forest. PMID- 2980378 TI - Is stress a developmental toxin? PMID- 2980379 TI - Nicotine and cotinine effects on development of two-cell mouse embryos in vitro. AB - The independent and interactive effects of nicotine and cotinine on the development of cultured two-cell embryos were investigated. Cultures were maintained for 120 h and developmental stages of embryos were scored after 72 h and at the termination of culture. Concentrations of nicotine at or below 0.5 mM, and concentrations of cotinine at or below 0.008 mM, did not adversely affect development. In addition, neither nicotine nor cotinine produced synergistic effects at higher concentrations at which both independently impaired development. These data show, therefore, that nicotine and its major metabolite, cotinine, significantly interfere with preimplantation development of mouse embryos only at concentrations far in excess of those anticipated to be present in the blood of an "average" smoker. Thus, we conclude that the well documented adverse effects of smoking during pregnancy are unlikely to be attributable to a direct effect of nicotine or cotinine on the preimplantation embryo. PMID- 2980380 TI - Effects of phytoestrogens on GnRH-induced luteinizing hormone secretion in ovariectomized rats. AB - Ovariectomized rats were pretreated with intravenous estradiol-17 beta (E2), coumestrol or genistein and were challenged 2 h later with GnRH (50 ng/kg BW, i.v.). Blood samples, drawn at the time of GnRH administration and 15 min afterwards, were assayed for luteinizing hormone (LH). While low-dose E2 pretreatment (10 ng/kg BW) significantly enhanced GnRH-induced LH release, high dose E2 pretreatment (1000 ng/kg BW) blocked the GnRH-induced rise. Intermediate dose E2 pretreatment (100 ng/kg BW) yielded a GnRH response that did not differ from that in the vehicle pretreatment group. At all doses studied, coumestrol pretreatment (10 ng/kg BW, 100 ng/kg BW, and 1000 ng/kg BW) appeared to diminish but not abrogate GnRH-induced LH release. While intermediate-dose genistein pretreatment (100 ng/kg BW) significantly enhanced GnRH-induced LH release, high dose genistein pretreatment (1000 ng/kg BW) yielded a GnRH response that did not differ from that in the vehicle pretreatment group. Both low-dose (10 ng/kg BW) and very high-dose (10,000 ng/kg BW) genistein pretreatment appeared to inhibit GnRH-induced LH release. The results demonstrate that acute exposure to phytoestrogens alters GnRH-induced LH release in ovariectomized rats. The dose response pattern of enhanced GnRH-induced LH release at lower pretreatment doses but inhibited GnRH-induced LH release at higher pretreatment doses was observed for E2 and genistein. The potency of genistein appears to be approximately 1/10 that of E2 in this system. Phytoestrogens acutely perturb reproductive neuroendocrine function. PMID- 2980381 TI - Lack of evidence of teratogenicity of benzodiazepine drugs in Hungary. AB - In order to investigate possible teratogenic effects of commonly used benzodiazepines (diazepam, chlordiazepoxide, nitrazepam) in Hungary, four approaches were used: 1. A retrospective case-control study of 630 cases with isolated cleft lip +/- cleft palate, 179 cases with isolated cleft palate, 392 cases of multiple congenital anomalies including cleft lip and/or cleft palate, and their matched control cases; 2. The Case-Control Surveillance System of Congenital Anomalies in Hungary, 1980 to 1984, involving 355 cases with isolated cleft palate, 417 cases with multiple congenital anomalies, and 186 cases with Down's syndrome (as positive controls). Benzodiazepines were taken by 14.9% of 11,073 control pregnant women studied; 3. A prospective study of 33 pregnant women attending the Counselling Clinic following ingestion of benzodiazepines during the first trimester of pregnancy; 4. An observational study involving 12 pregnant women who attempted suicide and one with accidental overdosage with benzodiazepines during pregnancy. None of these four approaches gave any indication of an association between facial clefting and in utero exposure to these substances. PMID- 2980382 TI - Effects of in-phase and out-of-phase ovulation induction on early mouse embryos. AB - The effects of ovulation induction on pre-implantation embryos (day 4 of gestation) were studied in mature cycling mice (the Jcl:ICR strain) using relatively small doses of pregnant mare's serum (PMS)/human chorionic gonadotropin (HCG) (2.5 or 5 IU each) given at two different stages (in-phase or out-of-phase to the estrous cycle). The proportion of degenerated embryos was increased and the development of the embryos was retarded in the treated groups, especially in the out-of-phase subgroups. Therefore, we propose that the use of PMS/HCG for ovulation induction should be more restricted in mouse reproductive experiments when mature females are used, and that, even when ovulation induction is used, a treatment synchronized to the spontaneous estrous cycle should be elected. PMID- 2980383 TI - Genital tract abnormalities in female rats exposed to diethylstilbestrol in utero. AB - Genital tract morphology in 14-month old female rats exposed prenatally to diethylstilbestrol (DES) was analyzed as part of an examination of the effects of transplacental exposure to DES on estrogen sensitive tissues. Pregnant Sprague Dawley rats were injected with sesame oil alone or with DES in sesame oil on days 10 and 13 of gestation (total dose 1.2 micrograms DES) or on days 15 and 18 (total dose 1.2 micrograms or 120 micrograms DES). Female offspring (9-15 per group) were sacrificed at 14 months of age. Effects of DES exposure varied with the dose given and with the stage of differentiation of the fetal tissues. In the ovaries of rats exposed to 120 micrograms of DES on days 15 and 18 of gestation, follicular elements were reduced and replaced by dense sheets of stromal cells; oophoritis was noted in five of nine rats. Hypercellularity of oviductal stroma was another common feature, as was suppurative salpingitis. Ovaries of rats exposed to 1.2 micrograms DES on days 10 and 13 of gestation were more likely to contain numerous corpora lutea than the other DES-exposed groups of controls. An increased incidence of benign uterine abnormalities was observed in DES-exposed offspring, including squamous metaplasia and suppurative endometritis. In the cervices of all nine rats exposed to 120 micrograms DES on days 15 and 18 of gestation, the epithelial surface showed a convoluted pattern, lined by stratified squamous and stratified cuboidal cells. Thus, prenatal exposure to DES, especially at the higher dose used, has long-term consequences on reproductive tract morphology in Sprague-Dawley rats. PMID- 2980384 TI - Placental abruption associated with cocaine use: case report. AB - The medical problems associated with cocaine ingestion, most notably cardiovascular side effects, have become evident as its use has become epidemic in the United States. Its effects on pregnancy and the developing fetus are similarly being recognized as more women of childbearing age are abusing cocaine. A case of placental abruption at 33 weeks' gestation following cocaine use is reported. A review of the pharmacodynamics, medical complications, teratogenic potential and effects in pregnancy is included. PMID- 2980385 TI - Use of primate folliculogenesis models in understanding human reproductive biology and applicability to toxicology. AB - The nonhuman primate reproductive system provides an excellent model for studying basic physiological processes applicable to humans. This article reviews hormonal observations and experimental manipulations useful in the evaluation of ovarian events in various stages of the reproductive life. As the need arises, primate reproductive toxicological studies may clarify questions relevant to human risk evaluations. Evaluation of reproductive toxicological observations may reveal biological parameters defining premature reproductive failure. PMID- 2980386 TI - Aging and responses to toxins in female reproductive functions. AB - Reproductive senescence in laboratory rodents has been well characterized using vaginal smear analysis, a relatively simple and inexpensive technique to monitor changes in estrous cyclicity. Although some differences exist, laboratory animal models are available to study most aspects of age-related changes in human reproductive physiology (4,13). Thus presumptive reproductive toxins can be tested on laboratory rodents using the qualitative and quantitative parameters for estrous cyclicity described here to assess damage. Before implementing such tests, however, certain limitations must be considered. Quantitative changes in estrous cycles, for example, cycle length distribution, which indicate more subtle impairments in reproductive function, require more refined data analysis. In particular, baseline data for at least one month is required before exposure to the presumptive toxin. In addition, the period of maximum cycling regularity is fairly short (three to four months duration) so long-term exposures to presumptive toxins would not be applicable. Longer term exposure (greater than or equal to six months) would be permissible if qualitative changes in cyclicity (i.e., acyclic vs. cyclic) were the dependent variable used to assess toxicity. Finally, if other parameters of reproductive function (e.g., fertility, litter size) are used to assess reproductive toxicity, we urge that cycling characteristics of the offspring be carefully monitored to assess possible cryptic damage to later neuroendocrine functions that occurred to the fetuses in utero. PMID- 2980387 TI - Studies of reproductive cyclicity: evaluation of computer modeling as a tool. AB - Assessment of toxic effects on human reproductive function using fetal health or teratogenic criteria presumes the conditions of parental fertility. Toxins that compromise fecundity through derangement of the menstrual cycle may require quite different models and criteria. Nonlinear dynamic interactions of the hormonal and morphological components of the menstrual cycle add to the difficulty of such studies. With appropriate mathematical models, computer simulation can provide a useful guide to the design and conduct of in vivo experiment. PMID- 2980388 TI - Old teratogens: are they still important? PMID- 2980389 TI - Nonprescription drugs as a source of aluminum, bismuth, and iodine during pregnancy. PMID- 2980390 TI - Long-term evaluation of the diethylstilbestrol (DES) syndrome in adult female rhesus monkeys (Macaca mulatta). AB - This study was designed to examine the transplacental effects of diethylstilbestrol (DES) in rhesus monkeys (Macaca mulatta). Nineteen pregnant females received an approximate human therapeutic dose of 1 mg/day DES beginning on gestational day 21 (Group I), day 100 (Group II), or day 130 (Group III) until term. Colposcopic examination and vaginal biopsies were performed in eight female offspring at 6- and 12-month intervals beginning at 3.5 years of age until death between 5 and 12 years of age. Gross and histological evidence of vaginal adenosis was observed in five of eight (62.5%) females. Adenosis, which occurred in all treatment groups, did not develop into neoplasia; rather, it underwent metaplasia and reversion to squamous epithelium, a frequent observation in human DES cases. The vaginal ridging and/or cervical hooding characteristic of the human syndrome was observed in all exposed females. Although menstrual cyclicity was not impaired, there appeared to be a lower pregnancy rate in treated animals compared to age-matched controls. This long-term evaluation of reproductive morphology and function in the rhesus monkey has provided a useful model for studying the history of benign vaginocervical abnormalities induced by prenatal DES treatment. PMID- 2980391 TI - The long-term effect of clomiphene citrate on vaginal epithelial differentiation in the mouse. AB - Clomiphene citrate, administered to the neonatal mouse during cervicovaginal epithelial differentiation, is associated with subsequent adenosis. Similar lack of neonatal transformation of columnar to squamous epithelium occurs after diethylstilbestrol. Mouse neonatally exposed to diethylstilbestrol have a high incidence of reproductive tract tumors in later life. In this study Balb/c neonates were exposed to clomiphene citrate, maintained with controls, and killed at 52, 56, and 60 weeks. Cervicovaginal histology was studied by light microscopy. In all control animals only mature squamous epithelium was seen (44/44). After clomiphene citrate, at 52 weeks, 3/10 animals had other than mature squamous epithelium identified. One of these genital tracts contained adenosis. At 56 weeks only mature squamous epithelium was seen (11/11), and at 60 weeks immature squamous epithelium was identified in 1 of 10 animals. In no study animal (0/31) was there any evidence of tumor development. This differs from the high incidence of reproductive tract tumor seen in the mouse after neonatal diethylstilbestrol exposure. PMID- 2980392 TI - Methylmercury effects on reproduction and offspring size at birth. AB - This article describes a study of the toxic, reproductive, and developmental effects of chronic methylmercury (MeHg) exposure in Macaca fascicularis monkeys. Adult and infant monkeys were studied using procedures to assess maternal and newborn blood Hg concentrations, menstrual cyclicity, conception rate, reproductive outcome, maternal toxicity, and offspring size at birth. Maternal intakes of 0, 50, 70, or 90 micrograms/kg/d MeHg hydroxide were studied. Maternal blood Hg concentrations reached equilibrium by 10 weeks of exposure. The half life of blood Hg for adult females ranged from 15 to 40 days (mean = d) and did not vary with dose. Maternal MeHg exposure did not affect the length of the menstrual cycle or the conception rate. Maternal MeHg exposure did significantly reduce the number of viable deliveries at blood Hg concentrations above 1.5 ppm. Maternal blood Hg concentrations at delivery were significantly lower than newborn concentrations. No effect of maternal MeHg exposure on offspring size at birth was observed. Maternal toxicity was related to blood Hg concentrations above 2.0 ppm following approximately one year of exposure. Results indicate that MeHg exposure can affect reproductive outcome at levels that do not cause overt toxicity. PMID- 2980394 TI - Effects of acetaminophen and nonsteroidal anti-inflammatory drugs on progesterone production by porcine granulosa cells in vitro. AB - We investigated the effects of a group of pharmaceutical agents commonly ingested by reproductive-aged women, acetaminophen and the nonsteroidal anti-inflammatory drugs (NSAID), on progesterone (P) production by cultures of highly differentiated porcine granulosa cells. These compounds were added to cultures over a dose range of 10(-8) to 10(-5) M and P, and cell protein was measured after 24 hours. P production was suppressed by acetaminophen, fenoprofen, and sulindac to a maximum of 81%, 76%, and 71% of control, respectively. P production was enhanced by butazolidin at all doses tested to a maximum of 140% of control. Granulosa cell protein was suppressed by butazolidin and salicylic acid to a maximum of 81% of controls. These data imply that acetaminophen and several NSAID have the potential for clinical reproductive toxicity with differing individual effects on reproductive tract tissues, suggesting further selective testing in vivo. PMID- 2980393 TI - Enhancement of methylmercury toxicity by L-cystine in cultured mouse blastocysts. AB - Expanded mouse blastocysts incubated with 1 to 2 microM methylmercury (MeHg) in modified Eagle's basal medium (BME + AA), which contains amino acids, collapsed and degenerated within 24 h. In contrast, blastocysts incubated with the same concentration of MeHg in egg culture medium (ECM), which does not contain amino acids, survived and remained expanded as control embryos did. By systematically omitting each BME amino acid from BME + AA and adding each BME amino acid to egg culture medium, we determined that L-cystine (0.5 mM in BME + AA) was the component of BME + AA that was responsible for the enhancement of the toxicity of MeHg. The shortest incubation time during which the cystine-enhanced MeHg toxicity became irreversible was 2 h, and the addition of any of the neutral BME amino acids (except threonine) or non-BME neutral amino acids (alanine, glycine, or serine) during the 2 h incubation eliminated or reduced the cystine-enhanced MeHg toxicity. Basic amino acids (except histidine) were less effective in protecting embryos: Glutamine and lysine reduced the toxic effect only slightly, and arginine had no effect. DL-buthionine sulfoximine (7.5 mM), a specific inhibitor of glutathione, also reduced cystine-enhanced MeHg toxicity. It therefore appears that cystine enhances MeHg toxicity indirectly, at least in part, by stimulating the synthesis of cellular glutathione, which may in turn enhance MeHg transport. In the absence of cystine, 10 microM MeHg (2 h incubation) was necessary to cause the collapse and degeneration of all blastocysts treated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980395 TI - Dose-response relationship of cadmium embryotoxicity in cultured mouse embryos. AB - Mouse embryos were exposed in vitro to 1.2 to 2.2 microM cadmium, and effects on embryotoxicity were examined after 39 h of culture. Teratogenic responses similar to in vivo were obtained at 1.2 to 2.2 microM with concomitant reduction in embryonic protein, while embryo deaths were increased from 13.8 to 93.3% at 2.0 to 2.2 microM. The response data of both teratogenicity and growth parameters, including embryonic protein, head length, crown-rump length, somite number, and protein and diameter of yolk sac, were acceptably fitted to a linear log-probit regression. These results suggest that (a) In chronic exposure conditions, the concentration of cadmium is a critical parameter in the manifestation of teratogenic potential, (b) as an estimation of interference in the growth of embryos, embryonic protein is one of the most sensitive endpoints while somite number is an insensitive criterion, and (c) a linear log-probit regression is applicable to the analyses of embryotoxicity data, including growth parameters in whole-embryo culture systems. PMID- 2980396 TI - How safe is safe for hormones? PMID- 2980397 TI - Case reports of individuals with oligospermia and methylene chloride exposures. AB - Between December 7, 1984 and June, 1986, 34 men with exposure to methylene chloride were evaluated at the Greater Cincinnati Occupational Health Center (GCOHC). Their primary complaint involved problems associated with central nervous system dysfunction. However, 8 of the 34 men complained of testicular, epididymal or lower abdominal pain (found on exam to be prostatic in origin) and had clinical histories consistent with infertility. Semen specimens from four volunteers were found to be in the subfertile or infertile range with regard to motility, morphology and sperm density. PMID- 2980398 TI - Developmental effects of progesterone and its derivatives. PMID- 2980399 TI - In vitro culture of postimplantation hamster embryos. AB - In vitro culture of intact rat and mouse embryos has been described extensively, but information on the culture of other species is sparse. The present study examined some culture requirements of early somite stage hamster embryos and assessed the embryotoxic effects of sodium salicylate (SS), a direct acting chemical and cyclophosphamide (CP), a proteratogen, on these embryos. Hamster embryos explanted on gestation days (GD) 8 and 9 were cultured in Waymouth's embryo-hepatocyte co-cultivation medium (WEHC), 70% McCoy's 5A medium-30% male rat serum (MMRS) or 100% male rat serum (MRS) for 24 hours under various oxygen concentrations. Embryos cultured GD 8 to 9 in the various media grew and differentiated much as they did in vivo, while embryos cultured GD 9 to 10 grew best in MMRS as compared to embryos at the same stage in vivo. Embryos exposed to SS in MMRS at concentrations of 250, 300, or 400 micrograms/ml showed dose related embryotoxicity which included CNS defects, absence of hind limb bud formation, and lack of axial rotation. Hamster embryos co-cultivated with pregnant hamster hepatocytes and treated with 2.5, 6.25 and 12.5 micrograms/ml of CP, showed dose-dependent toxicity when compared to co-cultivated controls. Hamster embryos develop extensively in culture over a 24 hour period. This system may therefore provide a valuable tool for evaluating the species differences of a variety of potential teratogens and embryotoxins and allow the comparison of these embryotoxic effects between rat, mouse and hamster during similar stages of organogenesis. PMID- 2980400 TI - Transport and metabolism of dexamethasone in the dually perfused human placenta. AB - Dexamethasone (DEX), a synthetic glucocorticoid, has been suggested to accelerate fetal lung maturation and is sometimes used clinically during premature labor for this purpose. These experiments were initiated to measure the rate of transfer and metabolism of DEX by the human placenta. Lobules from full term (38-41 weeks) human placentas, obtained immediately after delivery, were perfused on both maternal and fetal sides. Radiolabeled (14C) DEX and tritiated water were infused into the maternal artery. Samples were taken at timed intervals throughout the perfusion period from the maternal artery (MA), maternal vein (MV), fetal artery (FA) and fetal vein (FV). Transport was determined by liquid scintillation spectroscopy and metabolism of DEX was determined by reversed phase HPLC. Tritiated water reached equilibrium between the maternal and fetal circulations within 75 min of perfusion, while total 14C-labeled radioactivity attained equilibrium after 180 min. HPLC revealed one metabolite of DEX which co-migrated with an 11-ketobetamethasone standard. The concentration of metabolite at 10, 30, 120, and 240 min in the FV were 9, 22, 40, and 41 ng/ml, respectively, and in the MV were 15, 28, 46, and 48 ng/ml, respectively. However, the ratio of metabolite/DEX was greater at 10 min in the FV (0.39) than in the MV (0.08). These data suggest that DEX crosses the human term placenta, that an enzyme in the placenta metabolizes DEX into an 11-keto-metabolite, and that a greater metabolite:DEX ratio is found at earlier times in the fetal circulation than in the maternal circulation. PMID- 2980401 TI - Effects of acute and chronic exposure to cobalt on male reproduction in mice. AB - Chronic exposure of male mice to cobaltous chloride dramatically affected their reproductive potential, while acute administration had minimal effects. Acute exposure, followed by evaluation weekly over a 7-week period, revealed no significant changes in epididymal sperm concentration or testicular weight. However, small but significant decreases in fertility at weeks 2 and 3 of the study were observed. Sperm motility was depressed only during the first week of the study. In chronic studies, cobalt affected fertility in a time- and dose dependent manner. There was a decrease in testicular weight, epididymal sperm concentration, and fertility. Sperm motility was also depressed. Serum testosterone levels were dramatically increased in cobalt treated animals, while FSH and LH serum levels were normal. It appears that cobalt is directly or indirectly interfering with spermatogenesis and with local regulatory mechanisms in testosterone synthesis. PMID- 2980402 TI - Maternal toxicity and teratogenicity of tellurium dioxide in the Wistar rat: relationship to pair-feeding. AB - Tellurium dioxide (TeO2) induces hydrocephalus, edema, exophthalmia, ocular hemorrhage, umbilical hernia, undescended testes and small kidneys in day 20 Wistar rat fetuses when administered s.c. to pregnant dams from gestational day 15 to 19. At doses of 500 mumole/kg or greater, a 100% incidence of these findings and a reduction in maternal weight gain were observed. A pair-fed study at the dose of 500 mumole/kg of TeO2 was conducted to establish if the effects of tellurium were a result of a reduction in food intake or other maternal toxic responses. Two additional control groups of rats receiving tellurium or vehicle were fed ad libitum. After a comparable maternal weight gain from day 0 to 15, weight gain was significantly reduced in the treated groups and the pair-fed control. There was a reduction of fetal weight in the treated groups (p less than 0.01) and in the pair-fed control (p less than 0.02). There was a 100% incidence of the above anomalies in the litters of the two treated groups, but none in the pair-fed and control groups. No histological alterations other than a mild centrolobular fatty change in the liver were detected in the other organs from the tellurium exposed dams. Thus, tellurium induces both maternal toxicity and teratogenic effects in the rat where the teratogenicity is not mediated by alterations in the diet. PMID- 2980403 TI - Effect of "activated" cyclophosphamide on mouse oocyte in vitro fertilization and cleavage. AB - To investigate the involvement of oocytes in the mechanism of chemotherapy induced ovarian failure, ovulation was induced in mice using pregnant mares' serum gonadotropin and human chorionic gonadotropin. Oocytes (with cumulus) were incubated for 4 hours with "activated" cyclophosphamide, 4 hydroperoxycyclophosphamide (PCTX) at 0, 1, 10, 100, and 500 micrograms/mL. Oocytes were then washed, fertilized with sperm obtained from nontreated male proven breeders, and incubated for 4 days. "Activated" cyclophosphamide inhibited dissolution of the cumulus and reduced fertilization and early cleavage rates in a dose-related manner. The data demonstrate that exposure of oocytes to cyclophosphamide metabolites in vitro adversely affects oocyte function. Oocytes may be involved in the mechanism of cyclophosphamide-induced ovarian failure. PMID- 2980404 TI - An evaluation of the serum FSH as a biomarker for ovarian toxicity in the rat. AB - There is a need for a biomarker to evaluate numbers of oocytes in an ovary exposed to various exogenous toxicants. Experiments that had been previously reported were reviewed to assess the serum FSH as such a biomarker. Rats that had been exposed to direct ovarian radiation and direct ovarian radiation with cyclophosphamide were reviewed to ascertain the relationship of follicle numbers to the serum FSH. Under the conditions of reduced serum estrogen, linear regression analysis indicated that the serum FSH correlated best with the calculated volume of antral follicles. Receiver operating characteristic curves were calculated to determine that an elevation of the serum FSH two standard deviations above the mean had the greatest diagnostic power in predicting a significant loss in both the number of healthy follicles and the calculated volume of healthy antral follicles. The serum FSH as a biomarker appears to have utility as a measure of the ovarian content of healthy antral follicles rather than as a measure of the total number of oocytes alone. PMID- 2980405 TI - Delayed effects of doxorubicin on spermatogenesis and endocrine function in rats. AB - Doxorubicin was administered to adult male Wistar rats (1 mg/kg body weight, three times per week, for one, two, three, or four weeks) in order to examine testicular and reproductive endocrine toxicity 56 days after treatment. Doxorubicin treatment produced persistent dose-related reductions in testis, epididymis, and seminal vesicle weights, but did not alter ventral prostate weight. Testis and serum testosterone levels were not significantly affected by treatment, but serum LH was increased after treatment, and binding of iodinated hCG to testicular LH receptors was reduced. Serum FSH was elevated by the two lower total administered doses, but was not different from controls after treatment with the two higher total doses. There was clear histologic evidence of dose-dependent damage to the seminiferous tubules, which was reflected by decreased testicular and epididymal sperm content and by reductions in the stem cell survival index. These results indicate that doxorubicin produces significant and persistent damage to the endocrine and spermatogenic compartments of the testis. PMID- 2980406 TI - Permanent chondrification in the pelvis and occurrence of hernias in mice treated neonatally with tamoxifen. AB - Male and female C57BL/Tw mice were given 5 daily subcutaneous injections of 100 micrograms tamoxifen (Tx), starting on the day of birth (Tx mice). In untreated fetal mice on day 18 of gestation, the greater part of the pubic and ischial bones were cartilaginous. At more than 30 days of age, however, untreated mice showed completely calcified pelvic bone, whereas in age-matched Tx mice the greater part of the junctional regions in the pelvis remained cartilaginous. Treatment with Tx starting within 5 days of age caused bladder hernia with or without cecum hernia. The pubic ligament in Tx mice at ages of 30-540 days was markedly expanded as compared with that in age-matched controls. The permanent chondrification in the pelvis was found in all mice given Tx starting within 10 days of age. By contrast, neonatal treatments of mice with other antiestrogens, clomiphene and nafoxidine (100 micrograms/day), induced neither permanent chondrification in the pelvis nor expansion of the pubic ligament nor hernia. These findings suggest that Tx has a specific effect on the symphysis pubis and some junctional regions of the developing pelvis in mice when given neonatally. PMID- 2980407 TI - Evaluation of human serum as a culture medium for studies of rat embryonic development in vitro. AB - The normal growth of rat embryos cultured in human serum supplemented with glucose and 10% rat serum is of potential use in the assessment of teratogenic risk in man. Before this technique can be widely applied, it is necessary to know whether the stages of the menstrual cycle at which a serum sample is tested materially affect the results obtained. Moreover, in order to achieve reproducible conditions we have found it necessary to add a minimal amount of rat serum to the human serum used for culture, but unnecessarily high levels of rat serum supplementation could over-compensate for any growth factor deficiency in human serum. Here we report that culture of rat conceptuses gives similar results irrespective of whether human first, second, or third trimester pregnancy serum, postnatal serum, or serum at various stages of the menstrual cycle is used. We also report that addition of 2% rat serum supplement is sufficient to achieve reproducible rat embryonic growth and differentiation in human serum. PMID- 2980408 TI - Analysis of protective activity of N-acetylcysteine against teratogenicity of heavy metals. AB - N-acetylcysteine (NAC) is known to enhance the renal excretion of heavy metals. Therefore, we investigated whether the teratogenicity of metals (Hg, Cr, and Cd) can be ameliorated by NAC in mice. Contrary to our expectation, the incidence of congenital malformations produced by these metals was two to three times higher in the mice that were fed NAC (0.2% in the diet). The underlying mechanism or mechanisms are unknown and should be investigated. PMID- 2980409 TI - Clinical tests and statistics. PMID- 2980410 TI - Polychlorinated biphenyls and reproduction. PMID- 2980412 TI - The effect of "activated" cyclophosphamide on rat granulosa cells in vitro. AB - We investigated the mechanism of cyclophosphamide (CTX)-induced ovarian toxicity by studying the effect of an activated form, 4-hydroperoxycyclophosphamide (PCTX), on rat granulosa cells in vitro. Cells were obtained from PMSG-primed immature rats and incubated with PCTX at concentrations of 1, 10, 100, and 500 micrograms/mL. Ovine LH (10 ng/mL) was added in selected tubes. Cell viability before and after seven hours incubation was determined. Progesterone and prostaglandin E accumulation were measured by radioimmunoassay. Granulosa cell viability was significantly decreased at PCTX concentrations of 10 micrograms/mL or higher in a dose-related manner. PCTX at concentrations of 100 micrograms/mL and 500 micrograms/mL significantly decreased basal and LH-induced progesterone and prostaglandin E accumulation. The above findings demonstrate that cyclophosphamide metabolites decrease granulosa cell survival and function in vitro. These direct effects suggest a possible mechanism for CTX-induced premature ovarian failure. PMID- 2980411 TI - Teratogenicity of benzoic acid derivatives of retinoic acid in cultured mouse embryos. AB - Isotretinoin (13-cis-RA) is a human teratogen and mouse embryos exposed to 13-cis RA in vivo exhibit many of the same defects as humans. Early postimplantation mouse embryos exposed to 13-cis-RA in culture exhibit developmental alterations of the visceral arches, similar to those seen after in vivo exposure. Certain benzoic acid derivatives of retinoic acid have been shown to possess activity equal to or greater than retinoic acid in several in vitro systems. This study examines the teratogenic effects of some of these retinoids on mouse embryos in vitro. Day 8 CD-1 mouse embryos were cultured for 48 hours in the presence of these benzoic acid derivatives. With the exception of Ro-15-0778, all compounds produced visceral arch malformations similar to those seen in embryos exposed to 13-cis-RA, but at dramatically different effective concentrations. Extremely low concentrations of the retinoic acid-related compounds tested appear to have detrimental effects on embryonic development and these compounds may be poor candidates for therapeutic use. PMID- 2980414 TI - Dental education of expectant parents. PMID- 2980413 TI - Prevention from infancy through adolescence. PMID- 2980415 TI - Assessment of a 2% buffered glutaraldehyde solution in pulpotomized primary teeth of school children: a preliminary report. PMID- 2980416 TI - Formocresol pulpotomies in primary molars: a radiographic study in a pediatric dentistry practice. PMID- 2980417 TI - Frequency of crossbite in surgically treated cleft lip and/or palate children. PMID- 2980418 TI - Odontoma of a maxillary central incisor in a 10-year old black male. PMID- 2980419 TI - Papillon-Lefevre syndrome: pediatric dental management. PMID- 2980420 TI - Chlorhexidine: notes and comments. PMID- 2980421 TI - Dental sounding board ... excessive overjet and impending impaction of mandibular second bicuspids. PMID- 2980422 TI - Transcranial Doppler monitoring of blood flow in the middle cerebral artery during angiography and drug perfusion. AB - With the new technique of transcranial Doppler sonography it is possible to record transient changes of blood flow within the major basal cerebral arteries. The injection of saline and contrast medium into the internal carotid artery is followed immediately by a marked turbulence of flow within the middle cerebral artery depending on the amount and pressure of the injected material. In addition, the flow in the middle cerebral artery is increased by the injection of saline, but reduced by the injection of contrast medium for 3 to 4 seconds and finally increased for 8 to 10 seconds. No changes are seen on the contralateral side. The recordings of blood flow within the middle cerebral artery during 1.3 bis(2-chlorethyl)nitrosurea (BCNU) perfusion of glioblastoma recurrences via a flow-directed balloon catheter showed no obvious alterations during injections into the upper part of the carotid siphon, but a marked reduction of diastolic flow during injections into the middle cerebral artery. The reduction of flow may explain some of the complications of angiography. PMID- 2980423 TI - Magnetic resonance imaging of cerebral aneurysm. AB - Twenty patients with congenital cerebral aneurysms, studied on a 1.5 tesla MR unit, were also evaluated by angiography and/or computed tomography (CT) in order to establish the relative sensitivity of magnetic resonance imaging (MRI) in the demonstration of such aneurysms. Of the 27 aneurysms found on MRI, all but 4 were seen without the aid of the angiographic study. The four seen in retrospect included 3 out of 4 in the smallest size category (less than 5 x 5 mm) and one in next larger (5 x 5 mm to 10 x 10 mm). When more than one aneurysm was present or when an aneurysm and an arteriovenous malformation was present, MRI proved to be useful in demonstrating which bled. The presence of both subacute and acute clot identified the lesions that had bled. PMID- 2980424 TI - Non-invasive tests as an indicator of carotid stenosis. AB - Between 1979 and 1985, non-invasive testing was done on 896 carotid arteries prior to angiography. The non-invasive studies included B-scan ultrasound, Doppler, and oculoplethysmography. Estimates of per cent stenosis arrived at by the non-invasive reader was correlated with the degree of stenosis found at angiography. The Pearson correlation coefficient for agreement between the non invasive studies and angiography was 0.86. The mean absolute difference was 11 per cent. In 50 per cent of cases the results of the two examinations were identical. In 73 per cent of instances the non-invasive result was within 10 per cent of that of angiography. These results indicate that non-invasive testing can be highly accurate in estimating carotid stenosis and can be sustained over time. PMID- 2980425 TI - Magnetic resonance imaging of dural venous sinus invasion, occlusion and thrombosis. AB - Seventeen patients were found to have involvement of the dural venous sinuses on magnetic resonance imaging (MRI). In 7, there was tumor invasion through the dural coverings and in 10 thrombosis occurred as a result of various causes. MRI shows tumor in the sinus as a signal producing mass replacing the normal hypointensity of rapidly flowing blood. Disruption of the hypointense leaves of the enveloping dura is also well shown by MRI. Intravascular clotting, whether in the chemical state of deoxyhemoglobin or intra- or extracellular methemoglobin can be characterized by MRI operating at high field (1.5 tesla). MRI is now the non-invasive diagnostic modality of choice in evaluation of the dural venous sinuses. PMID- 2980426 TI - Focal recurrent hemorrhage on magnetic resonance at 0.5 tesla. An aid to the diagnosis of cryptic cerebral vascular malformations. AB - The MR appearance of focal recurrent hemorrhage is characteristic and highly suggestive of an underlying cryptic vascular malformation. We reviewed the MR findings in 14 cases (3 biopsy proven), 9 in the brainstem and 5 supratentorial. CT in all cases demonstrated the lesion to be hyperdense before contrast with minimal to moderate enhancement post contrast. The angiograms were negative in all 14 cases demonstrating mass effect in some but no blush or neovascularity. MR showed focal hyperintensities on both short (T1) and long (T2) TR sequences surrounded by a well-circumscribed hypointense rim particularly on the long TR sequence. Focal central hyperintensity is felt to be secondary to the presence of methemoglobin and indicates relatively recent hemorrhage. In our experience hyperintensity in a solitary hemorrhage usually resolves in 4-6 weeks. In 8 cases serial MR studies over 2-3 years demonstrated evidence of repeated hemorrhage as manifested by increases in size and persistent or new hyperintensity. The intense rim due to a short T2 did not correlate with vessels angiographically nor with calcification on the pre-contrast CT and, therefore, does not represent the vascular malformation itself but rather some portion of the hematoma, possibly the hemosiderin laden capsule. Using CT and angiography alone it is difficult to differentiate cryptic arteriovenous malformations from a neoplasm particularly in the brainstem. MR's specific configuration of focal recurrent hemorrhage can make a more definitive diagnosis at 0.5 T as well as at 1.5 T. PMID- 2980427 TI - Regional cerebral blood flow assessment prior to balloon detachment in the treatment of intracranial giant aneurysms. AB - Three patients with giant aneurysms (2 internal carotid and 1 anterior communicating) were treated by internal carotid occlusion with a detachable balloon. 133Xe regional cerebral blood flow (rCBF) was performed on each patient on admission. Due to low CBF, one patient received a superficial temporal artery- middle cerebral artery (STA-MCA) bypass. The rCBF was repeated when the balloon was inflated in the internal carotid prior to detachment of the balloon. All three patients were discharged within one week with no neurologic deficit. The rCBF assessment appears useful to decide which patient will tolerate acute balloon occlusion of the internal carotid and to help select patients who will need an extra-cranial-intracranial (EC-IC) bypass to avoid ischemic complication. PMID- 2980428 TI - Common carotid artery occlusion. Role of non-invasive and angiographic analysis. AB - Doppler and B-scan imaging, periorbital directional Doppler, and occuloplethysmography correctly identified common carotid occlusion in three of four consecutive patients with atherosclerosis. High-grade common carotid stenosis in the fourth patient could not be distinguished from occlusion on the non-invasive tests. Occlusion of the internal carotid artery in one patient and patency of that artery in the other three were evident non-invasively. Modified angiography agreed with non-invasive tests in each of three patients studied. Intra-arterial digital subtraction angiography in the fourth patient suggested but could not confirm patency of the internal carotid artery indicated by non invasive criteria. PMID- 2980429 TI - Aneurysm of the circle of Willis. Comparison between computed tomography and angiography. AB - The authors report their experience in 13 patients with unruptured non-giant aneurysms diagnosed at computed tomography (CT), who were without clinical deficits but who complained of chronic headache or cranial nerve palsy and had other members in their families with a cerebral vascular malformation. Screening with contrast-enhanced CT is discussed with respect to certain social groups. Surgical management before rupture of aneurysms could be envisaged in view of the bad prognosis when rupture has occurred. PMID- 2980430 TI - 'Open angioplasty' in carotid artery obstructions. AB - A new method has been introduced for the treatment of highly located or long segment stenoses or obstructions of the internal carotid artery not accessible by conventional carotid-endarterectomy. Following conventional carotid endarterectomy a balloon catheter is passed through the incised carotid artery as far as the stenosis and then inflated several times. Possibly dislodged plaque material can be washed out by the retrograde blood stream. The results in 30 cases, with angiopraphic, Doppler sonographic and neurologic follow-up examinations, are reported. It was possible to achieve a distinct additional cerebral blood supply through the recanalized internal carotid artery by this method, without a severe complication rate. PMID- 2980431 TI - Persisting abnormal embryonic vessels in intracranial arteriovenous malformations. AB - The study of arterial and venous anatomy of hemispheric cerebral arteriovenous malformations (AVMs) shows that, beside occlusion, rerouting of the flow, and collateral supply and drainage, the arterial and venous patterns are normal. This can be explained by the fact that the cerebral hemispheres develop late relative to other neural structures, and their fetal vessels become in fact the permanent vessels. This normality indicates that the AVMs develop after the vascular pattern has been determined. Vein of Galen aneurysms, in contrast, demonstrate occasionally an abnormal arterial pattern over the hemisphere, and retention of an embryonic vein, the median prosencephalic vein. This indicates that the fistula occurred at a much earlier stage, actually when the choroid plexuses first developed, about the end of the second month. The occurrence of choroid AVMs similar to the vein of Galen aneurysms, but drained by a normal internal cerebral vein, indicates that the time when the choroid drainage is taken over the internal cerebral vein, represents another critical period. PMID- 2980432 TI - Encephalic microvascular abnormalities in hypertension. A light and X-ray microscopic study. AB - Brain and cervical spinal cord removed at autopsy from hypertensive humans have been subjected to 4 levels of microscopic study, 3 being first histochemically prepared so as to stain the arterial and capillary system. Thick (100 microns) sections for light microscopy and thicker (500-1000 microns) for x-ray microscopy facilitate the study of long, complex anatomic units. Arterial and arteriolar microaneurysms have not been found. In addition to the usual age related vascular changes in normal people we have found: moniliform capillary dilatations, focal segments of arteriolar narrowing, etat pre-crible, and lacunar infarcts. Few changes have been encountered in the corpus callosum. While the segmental arteriolar narrowing occurs in the cortex as well as deep white matter, the effects of resultant poor perfusion might be expected to be more profound in the deep white matter. PMID- 2980433 TI - Detection of carotid thrombi with indium-111 platelet scintigraphy. AB - In order to prove that platelet scintigraphy (PSC) is able to detect carotid thrombi formations, we performed PSC in 15 patients directly before or immediately after a percutaneous carotid angiography. PSC was successful in demonstrating iatrogenic fresh carotid thrombi in 13 out of 15 cases. Out of 53 patients with cerebrovascular disease and carotid stenosis a pathologic platelet accumulation was seen in 39 carotid arteries, 31 on the symptomatic side; more often in slight- and middle-sized stenoses than in high degree stenoses. We assume that in these cases PSC detected carotid thrombi which caused arterio arterial emboli. PMID- 2980434 TI - Is computed tomography really effective in the early detection of brain tumors? A cooperative study. AB - A cooperative study based on 1,112 patients was undertaken in order to ascertain whether computed tomography (CT) allows earlier diagnosis of brain tumors or not. The patients were divided into 2 groups. In the first group (pre-CT period; 552 patients) diagnosis was made without CT, in the second (CT period; 560 patients) with CT. Duration of symptoms, assumed as the interval between onset and definite diagnosis, was the main parameter considered. The most relevant results were: a) during the CT period a significant decrease in duration of symptoms before diagnosis was observed for meningiomas (from 21 to 13 months) and low-grade gliomas (from 16 to 9 months), while no significant change was recorded for high grade tumors (from 4.6 to 3.5 months); b) during the CT period the figure for benign tumors operated upon rose to 44 per cent (pre-CT: 25%) balanced by a marked decrease in surgery for malignancies (from 43% to 23%); c) better indications for surgery were associated with a significant reduction in cases operated upon during the CT period (72% versus 79%, p less than 0.01). PMID- 2980435 TI - Primary intracranial lymphoma. An important differential diagnosis in magnetic resonance imaging of the brain. AB - The magnetic resonance (MR) appearances are described in 6 pathologically verified cases of primary intracranial lymphoma, and comparison is made with the appearances on computed tomography (CT). In 5 cases, atypical CT appearances were seen and in these cases more extensive and multiple abnormalities were demonstrated by MRI. The importance of differentiating this condition from other pathologies affecting the white matter is discussed. PMID- 2980436 TI - Magnetic resonance imaging in intracranial gliomas. Comparison with computed tomography and serial stereotactic biopsy. AB - Findings at magnetic resonance imaging (MRI) and computed tomography (CT) where reviewed in 53 surgically confirmed cases with glioma. Astrocytomas showed a constant MR signal pattern in 87 per cent and glioblastomas in 80 per cent. This pattern was found useful when it was correlated with CT. MRI was also found to be of value in planning stereotactic biopsy and in confirming the trajectory. PMID- 2980437 TI - Magnetic resonance imaging does not define tumor boundaries. AB - To define tumor boundaries prior to radiation therapy, 13 patients with intracranial glial neoplasms had tumor biopsies using magnetic resonance imaging (MRI)--computed tomography (CT) stereotactic technique. Precise histologic sampling of the lesion volume was correlated with the MRI and CT-defined 'lesion'. Tumor boundaries extended beyond the CT or MRI margin in 4 of 4 cases of glioblastoma, 1 of 3 cases of anaplastic astrocytoma, and 1 of 6 cases of well differentiated astrocytoma. While stereotactic MRI is superior to stereotactic CT in the visualization of glial neoplasms, accurate tumor biopsy is mandatory to define both tumor type and margins before pre-operative radiation therapy. PMID- 2980438 TI - Positron emission tomography with 11C-methionine of intracranial tumours compared with histology of multiple biopsies. AB - Thirty-eight patients with suspected supratentorial gliomas have been investigated with positron emission tomography (PET) using 11C-methionine and computed tomography (CT) before and after intravenous injection of contrast medium. Every examination was performed with the head fixed in a plastic helmet and a baseplate to guarantee that the slices were corresponding in the different examinations and from time to time. After the examinations multiple stereotactic biopsies were performed with the biopsy instrument mounted on the baseplate. Accumulation of methionine was compared with normal brain in 32 cases of tumours and ordinary or decreased accumulation in 4 cases of tumours. In 22 cases with increased accumulation of methionine the extension of the tumour judged by PET corresponded with that of histology. In 5 cases tumour cells were found outside the area with pathologic methionine uptake. In 5 patients there were areas with increased methionine accumulation where no tumour cells were found. In 24 cases PET using methionine was more accurate than CT. PMID- 2980439 TI - Tumour delineation with magnetic resonance imaging in gliomas. A comparison with positron emission tomography and computed tomography. AB - In ten patients with histopathologically confirmed low-grade (n = 4) and high grade (n = 6) cerebral gliomas, the ability of magnetic resonance imaging (MRI) to delineate these tumours was examined. Spin echo (SE) technique with dual and multiple echoes and subtraction of images recorded with multiple echo sequences was applied. Computed tomography (CT) and positron emission tomography (PET) with 11C-L-methionine were used as reference methods. In high-grade gliomas multiple echo sequences and subtractions delineated the tumours better than ordinary SE sequences and equally well as CT, but slightly less well than PET. In low-grade tumours, the tumour delineation was better than with CT and comparable to that obtained with the present PET method. Multiple echo sequences and subtraction increase the reliability of MRI in delineating cerebral gliomas as compared with standard SE sequences. PMID- 2980440 TI - Intraoperative ultrasound as guide for neurosurgical endoscopic procedures. AB - A high-tec ultrasound-guided device for neurosurgical endoscopy is described. Intraoperative ultrasound is used to bring a 6 mm rigid endoscope tube to a wanted target area. Repeated ultrasound imaging control provides a survey of intracerebral structures surrounding the endoscopically approached operation cavity. The endoscope harbours a suction irrigation system for aspiration of lobar intracerebral, intraventricular and cerebellar hematomas. Moreover, a 0.6 mm Neodym YAG laser tube and various microinstruments allow the biopsy and resection as well as laser irradiation of ventricular and cystic intracerebral tumors. A miniaturised TV-camera attached to the endoscope's ocular allows continuous visual control over surgical procedures. This procedure has been experienced in 105 patients as a low risk intervention with minimal surgical trauma. PMID- 2980441 TI - Magnetic resonance imaging of clival chordomas. AB - Chordoma of the clivus is one of the skull base tumors which originate from the remains of the notochord. The magnetic resonance findings in six patients are reported. The tumors showed slightly low intensity on T1 weighted images, but high intensity on T2 weighted images. The localization of the tumor was easily recognized by the disappearance of the normal signal in the clivus. Compression of the pons and medulla oblongata was readily diagnosed on sagittal images. A normal pituitary gland was demonstrated in all cases. Magnetic resonance imaging was useful for the diagnosis and for the surgical treatment of these chordomas. PMID- 2980442 TI - Magnetic resonance angiography in the head and neck. AB - Magnetic resonance angiography (MRA) provides a means to visualize vessel structure without the use of contrast agents and consequently without the risks and discomfort associated with contrast media. We have developed a flow encoding procedure which exploits the spatial dependence in the phase of spin magnetization generated by a gradient pulse. This technique yields image information for only those spins having macroscopic motion (i.e. flowing blood). Excellent suppression of non-moving spins can be obtained despite the overwhelming abundance of stationary spins. Data are presented in a projection format similar to that of traditional subtraction angiography. PMID- 2980443 TI - The relative contribution of magnetic resonance imaging to the assessment and differential diagnosis of cerebellopontine angle mass lesions. AB - Forty-nine patients with a space occupying lesion in the cerebellopontine angle cistern were examined by computed tomography (CT), magnetic imaging (MRI) and, where appropriate, conventional neuroradiologic techniques. The presence of an intracanalicular extension and a high signal on T2 weighted sequences were typical of acoustic neuroma and allowed separation from meningioma. Contrast enhancement with gadolinium DTPA was particularly valuable in recurrent acoustic neuroma. Typically epidermoids had long T1 and T2 values. Sequences which highlight flow allow vascular lesions to be identified without the use of contrast media. In separating the several pathologies MRI was superior to CT and also gave information on tissue composition in acoustic neuroma. PMID- 2980444 TI - High field magnetic resonance evaluation of acoustic neurinomas. AB - Eight acoustic neurinomas were studied with magnetic resonance imaging (MRI) at 1.5 tesla utilizing spin-echo T1 and T2 weighted images (WI). T1 WI identified the tumors best and provided more exacting anatomic relationships particularly for small lesions than did T2 WI. Intracanalicular tumors are well defined. All tumors were hypointense on T1 WI but were usually heterogeneous and had variable intensity on T2 WI. Several lesions became obscured on T2 WI because of isointensity with the adjacent cerebellum or CSF. High signal to noise ratio at 1.5 tesla permits thin section imaging (3 mm) at relatively short imaging times. T1 MRI was considerably superior to CT for the detection and anatomic definition of acoustic neurinomas. PMID- 2980445 TI - Correlation between MR signal intensity and histologic findings in neurinomas and meningiomas of the brain. AB - The signal intensity of magnetic resonance imaging (MRI) was compared with the histologic findings in 20 cases of neurinoma and 18 cases of meningioma. The apparatus was a Hitachi G-10 unit (0.15 T). The pulse sequences were SR (500/32), SE (1200/60), IR (1400/400). The intensity of tumor signal, as compared with that of normal white matter signal, was divided into 9 grades. In neurinomas, the signal intensity appeared markedly low with IR and markedly high with SE sequence in the Antoni B type, while it was slightly low with IR and minimally to markedly high with SE sequence in the Antoni A type, and intermediate between Antoni A type and B type in the mixed type. In meningiomas, it appeared moderately to slightly low with IR and slightly to moderately high with SE sequence in the meningotheliomatous type, minimally low with IR and minimally to slightly high with SE sequence in the fibrous type, and intermediate as compared with the above two in the transitional type. The angiomatous type was characterized by a low intensity on SR and IR images and an iso to high intensity on SE images, while the psammomatous type showed a low intensity with all of the sequences used. We clarified that the signal intensity on MRI varied among different histologic types of neurinoma or meningioma, and correlated differences in the intensity of signals on MRI to that of histologic characteristics, such as size and shape of cells, intercellular material, intracellular biologic polymers, area of membrane surface, blood flow and calcified matter. PMID- 2980446 TI - Radiologic assessment of acoustic neuroma in the elderly. Is no treatment good treatment? AB - Twenty-three elderly patients with acoustic neuroma have been followed conservatively (no treatment) for a mean of 3.8 years. Five patients subsequently required ventricular shunting. Two patients required partial excision of their tumors (one previously treated by shunting). Eleven patients who have required no treatment have no significant complaints. There was significant variation in tumor growth rates making predictability for surgical intervention hazardous. A mean growth rate of 0.22 cm/year was noted. Our statistics suggest tumor removal in healthy patients with larger tumors at time of presentation; in other patients, conservative follow-up may be more appropriate. PMID- 2980447 TI - Magnetic resonance and computed tomographic imaging of mass lesions of the cerebral ventricles. A prospective comparative study. AB - This study was undertaken to investigate the relative merits of magnetic resonance (MR) imaging and computed tomography (CT) in the evaluation of primary lesions of the cerebral ventricles. Since 1982, 14 patients, 7 males and 7 females, aged 8 to 68 years were diagnosed as harboring ventricular mass lesions. Eight masses were located in the lateral, 5 in the third, and one in the fourth ventricle. The diagnostic contribution and relative merits of two imaging modalities were compared in each case. In 57 per cent of cases the CT scan was of superior diagnostic value, and in 21 per cent of the patients MR imaging proved to be either more sensitive or specific than CT scanning. The availability of intravenous contrast agents for CT and not MR, as well as the capability of CT to definitely identify lesion calcification, accounted for the better performance of CT scanning in this series of patients. PMID- 2980448 TI - Accuracy of computed tomography in the diagnosis of herpes simplex encephalitis. AB - The accuracy of computed tomography (CT) in the diagnosis of herpes simplex encephalitis (HSE) was assessed in 121 patients who during a 2 1/2 year period entered a prospective Swedish joint study with participation of six University Centres. The patients presented with symptoms and signs of febrile focal encephalopathy. The age ranged from 1 month to 76 years (mean 37.3 years). Only 6 were infants less than one year old. HSE was diagnosed in 50 patients by the demonstration of intrathecal HSV antibody production and/or by HSV isolation or antigen detection in brain tissue specimens. A total of 308 CT examinations--the majority performed during the first 5 days after onset of CNS symptoms--were evaluated under blind conditions. The correct diagnosis of HSE was usually suggested within 5 days after onset of neurologic symptoms yielding a sensitivity of 0.73 and a specificity of 0.89. Predominant location of the HSE lesions was the temporal lobes (88%), which rarely were involved in the non-HSE group (11%). Haemorrhage was a rare finding (12% ) and enhancement after intravenous contrast administration was insignificant. Repeat examinations further increased accuracy. It is concluded that the good reliability of high resolution CT, further improvement of immunologic techniques and the advent of new atoxic antiviral drugs all are factors which may in the future obviate the need for brain biopsy. PMID- 2980449 TI - Imaging studies in neonatal herpes simplex encephalitis. AB - Herpes simplex encephalitis (HSE) is the most common form of sporadic encephalitis in man and usually results in death or permanent neurologic deficit. Neonatal HSE does not show the characteristic anatomic localization in the temporal lobe observed in adults and in older children which suggests that the disease process is different in this age group. The abnormalities observed with radionuclide brain imaging and computed tomography (CT) are more diffuse and bilateral. The clinical outcome of the disease is highly dependent on early institution of therapy. Therefore, early diagnosis is critical. Knowledge of neonatal HSE imaging patterns may alert the clinician to the proper diagnosis. PMID- 2980451 TI - Cortical asymmetry on magnetic resonance imaging related to unilateral hemispheric function. A preliminary investigation. AB - Cortical brain R/L asymmetry was assessed by spin echo lateral sagittal sections on magnetic resonance imaging (MRI) classified by visual comparison and by measuring the length of the parietal operculum and the inclination of the sylvian fissure. The results were related to handedness, selected as a lateralized function. The results indicate that assessment of the cortical surface by sagittal MRI is feasible. Comparison between images from two imaging systems (40 hemispheres: 0.15 T/0.75 and 1 cm section thickness versus 4 hemispheres: 0.5 T/0.5 cm section thickness) indicates a minimum standard for this approach on a large scale to be set by the more powerful system, mainly dependent on image noise and volume averaging factors. PMID- 2980450 TI - The hippocampal formation in coronal plane. PMID- 2980452 TI - Left-right asymmetry in the temporal and parietal region based on the medullary pattern of cerebral white matter. AB - Left-to-right differences in cerebral structures of the posterior temporal and inferior parietal region were investigated in CT examinations from 500 cases, by anatomic analysis based on the medullary branches of cerebral white matter. On the left side, the planum temporale and middle temporal gyrus were positioned more posteriorly, the intraparietal sulcus was deeper and lower, and the folding of the inferior parietal lobule was more extensive. The lateralization of the planum temporale diminished gradually from 19 years of age downwards. An anatomic analysis of the medullary branches succeeded in revealing the cerebral asymmetry and is expected to be useful in magnetic resonance imaging. PMID- 2980453 TI - Percutaneous transluminal angioscopy. A new imaging technique for diagnosis of extracranial arterial disease. AB - Angiography and ultrasound techniques are well established methods for the detection of extracranial vascular disease, but they only allow indirect demonstration of changes in the vessel wall. The development of an ultra-thin fiberscope allows direct demonstration of the pathogenic process in the vessel lumina by angioscopy. The percutaneous transfemoral approach for carotid angiography enables the combination of both methods. After development of an appropriate catheter system we investigated the value of this method producing definite lesions by vascular surgery in adult dogs followed by repeated examinations using angioscopy. Vessel wall alterations such as stalagmite-shaped lesions, subintimal bleeding and thrombotic layers, plaques, ulcerations and postoperative intimal bridging or ablation, which could not be identified by angiography have been analysed with high resolution of details using angioscopy with a video monitoring technique. PMID- 2980454 TI - Magnetic resonance imaging of the facial nerve. Normal anatomy and pathology. AB - Magnetic resonance imaging with surface coils permits visualization of the facial nerve from within the brainstem and continuing through a major portion of the parotid gland. Diagnostic capabilities have been expanded to include white matter disease that affects the tracts of the facial nerve within the brainstem and tumors of the parotid gland that involve the facial nerve trunks after the nerve exists from the stylomastoid foramen. PMID- 2980455 TI - Multivolumetric analysis of CT scans on patients with glioma. AB - Brain tumor volumes in patients having multimodal therapy for cerebral gliomas were calculated using graphic methods and Simpsonian integration. Volume assessment calculations differed from the clinical assessment of the patient significantly in some cases. These calculations may be useful in on-going tumor therapy and should be used as a true scientific end point in brain tumor treatment protocols. PMID- 2980456 TI - Intra-arterial chemotherapy with ACNU for the treatment of glioblastoma. Preliminary experience. AB - In November 1985, we started a study of intra-arterial (i-a) chemotherapy with ACNU for the treatment of glioblastomas of the central nervous system: 19 patients with histologically proved glioblastoma and recurrent, progressive or newly diagnosed disease were entered. Five patients were treated three times. We observed reduction of mass effect, of neovascularization, and of contrast enhancement. As to the time of survival, or follow-up is too short to allow definite conclusions. The quality of life in those patients who received several courses of chemotherapy, did not deteriorate as evidenced by a constant Karnofsky performance rating score. Systemic complications of i-a chemotherapy were negligible. However, some severe side-effects were seen: cerebral ischemia in two cases, and amaurosis in one. Direct ACNU related neurotoxicity was not seen to the present. According to these preliminary results, we feel encouraged to treat further patients with i-a chemotherapy, particularly in view of the disappointing results of several other treatment modalities in the management of glioblastomas of the brain. PMID- 2980458 TI - Radiation necrosis of the brain and radiation-induced cerebrovasculopathy. AB - Thirteen patients with radiation necrosis and 6 patients with occlusive radiation induced cerebrovasculopathy were reviewed. Computed tomography (CT) demonstrated low attenuation with a mass effect and irregular contrast enhancement in the radiation necrosis. The mass effect and the contrast enhancement decreased gradually. Angiography showed an avascular mass, generalized narrowing of the ipsilateral arteries and slow blood circulation in 3 cases. Radiation-induced vasculopathy appeared as localized stenosis and irregular walls of the arteries within the irradiated area. In cases with radiation vasculopathy accompanied by sufficient collateral blood flow brain damage was rarely demonstrated by CT. In patients with radiation-induced vasculopathy the lesion was not associated with signs of radiation necrosis. PMID- 2980457 TI - Blood brain barrier modification and chemotherapy. Interventional neuroradiology in the treatment of malignant gliomas. AB - Postoperative interventional neuroradiology was performed in patients with malignant gliomas to increase target efficacy of chemotherapy. In 8 glioma patients the blood brain or blood tumor barrier was reversibly opened by intraarterial injection of hyperosmolar fluid (Mannitol 25%). One additional patient had primary lymphoma of the central nervous system. During barrier modification chemotherapeutic agents were applied intraarterially and intravenously. A total of 22 blood brain barrier modification procedures have been carried out until now, ranging from one to five per patient. A presently continuing tumor regression or tumor progression free intervals have been noted in 5 patients. Therapeutic effects are being evaluated from repeated computed tomography and single photon emission computed tomography examinations. PMID- 2980459 TI - Radiologic manifestations of delayed radiation necrosis of the brain. AB - One of the most effective treatment modalities for tumors of the head and neck has been radiation therapy. In a small number of cases, delayed radiation necrosis of the brain is a dreaded and devastating complication of radiation therapy. The imaging studies (radionuclide brain scan, cerebral angiography, computed tomography and magnetic resonance imaging) in 12 cases of histologically proven, delayed radiation necrosis of the brain are analysed. While both computed tomography and magnetic resonance imaging can depict radiation necrosis with great sensitivity, however, the ability of these imaging techniques to differentiate between radiation necrosis and recurrent or residual brain tumor is low. PMID- 2980460 TI - Soft cervical disc herniations. AB - The authors report their findings in a series of 51 patients suffering from typical cervical radiculopathy. With the aid of the CT scanner and intravenous injection of a contrast medium, the quality and resolution of the resulting images allowed neurosurgical intervention in 42 patients in whom the symptomatology alone was not sufficiently informative to allow successful treatment. In most cases the CT images allowed an accurate diagnosis and revealed a free fragment that had torn the common posterior vertebra ligament. PMID- 2980461 TI - Magnetic resonance imaging in syringomyelia. AB - The aim of the study was to assess the reliability of magnetic resonance imaging (MRI) in the detection of syrinx cavities and to determine the proportion of cavities associated with Chiari malformation. MRI was performed in over 120 patients with neurologic deficits referable to the cranio-cervical junction. Forty-one patients were also evaluated by CT myelography (CTM). CTM demonstrated appearances consistent with intramedullary cavitation in 14 patients. MRI detected 11 of these but in 3 the appearances, including cord size, were normal. In the entire study 27 syrinx cavities were demonstrated by MRI, 8 associated with trauma, posterior fossa or spinal tumours, or spinal dysraphism. Of the remaining 19, tonsillar ectopia was present in 10 (53%). It is concluded that a lack of concordance between MRI and CTM exists in the detection of small cavities in the normal sized cord, and that syringomyelia without evidence of tonsillar ectopia is identifiable by MRI more frequently than might be anticipated. PMID- 2980462 TI - Carotid bifurcation imaging model for more accurately comparing imaging techniques. AB - For comparing current and presubably forthcoming imaging modalities, a carotid bifurcation model was made from cadaveric specimens. Perfusing and pulsing the immersed common carotid artery and its proximal branches via a Harvard pump simulated clinical imaging conditions. Film-screen (F-S) and digital subtraction (DS) angiography, computed tomography (CT), ultrasound and magnetic resonance imaging (MRI) were compared. For CT and MRI, scanning parameters such as slice thickness, degree of overlapping, amount of contrast medium needed, scanning mode and multiplanar and three-dimensional techniques enabled enhancing the capacity for CT in the clinical setting. Direct axial CT proved to be most accurate for assessing the contours and magnitude of carotid narrowing. Nonetheless, these serial segments were not readily compared with F-S and DS angiographic full length images. The use of multiplanar reconstruction (MPR) and three-dimensional (3-D) CT achieved this and furthermore showed the external contour of the diseased segment. Concerning carotid ulceration, our carotid model study showed CT to be equally accurate with DS and superior to F-S angiography. However, in our clinical study of 34 carotid arteries in 17 patients ulcerations were equally well identified by CT and angiography but DS angiography proved superior in identifying ulcers not seen with CT. Perhaps this discrepancy is explained by the clinical routine of attaining multiple fluoroscopically positioned views of the common carotid bifurcations in DS catheter angiography unlike the complexity of attaining optimal views of tortuous vessels on CT. PMID- 2980463 TI - Magnetic resonance imaging in the differentiation of spinal cord tumours and hydromyelia. AB - Twenty-two cases of intramedullary spinal tumours with cystic parts and 22 cases of hydrodynamic syringomyelia were classified according to criteria such as age at onset, duration of symptoms, nature of neurologic symptoms and a number of imaging parameters. The criteria used proved to be helpful in making the correct diagnosis. PMID- 2980464 TI - Radiologic demonstration of syringobulbia. Report of 8 cases. AB - Out of 75 syringomyelia cases, 8 had a syringobulbic extension. The cavity was demonstrated by magnetic resonance imaging (3 cases), syringography (3 cases), ventriculography (2 cases), and myelo-CT (2 cases). The cavities could be separated into 2 groups. In the first (5 cases) the cavity extended from the floor of the fourth ventricle, anterolaterally (3 cases) or anteriorly in the midline (2 cases). The communication with the fourth ventricle was usually wide. In the second group (3 cases), the syringobulbic cavity was located anteriorly; 2 of the 3 had no communication with the fourth ventricle and had an exceptional cranial extension (syringocephalia). In selected cases, invasive studies may help to recognize the connections of the cavities, and thus be useful in planning the surgical treatment. PMID- 2980465 TI - Subtle cord magnetic resonance changes in cervical myelomalacia. AB - After spinal cord injury cystic lesions of different types are known to develop. However, in a large group of patients radiologic examinations have not revealed abnormalities in spite of neurologic deficit symptoms. Magnetic resonance imaging in 6 such patients demonstrated subtle changes in the cervical spinal cord, confirming the diagnosis of myelomalacia. PMID- 2980466 TI - Radiologic studies of cerebrospinal fluid pathways in experimental hydrocephalus hydrosyringomyelia. AB - The progress of experimental hydrocephalus-hydromyelia in cats is analyzed in the present study. The animals were rendered hydrocephalic by intracisternal injections of kaolin or by operatively occluding the lateral apertures. Changes in the cerebrospinal fluid (CSF) spaces were monitored by ventriculography or computed tomography. The progress of hydrocephalus in both groups can be divided into two stages: 1) A phase of raised intracranial pressure precedes the dilatation of the central canal, and 2) a progressive phase follows the dilatation of the central canal. The canal acts as a kind of natural bypass between the ventricles and the subarachnoid spaces. Although the spontaneous shunt, the central canal, becomes dilated and pressures drop to near normal values, the disease is apparently not brought to a complete halt. Our observations suggest that the disturbance in CSF flow dynamics is an important factor causing hydrocephalus to remain progressive. PMID- 2980467 TI - Examination of epidural tumors of the spinal canal. Comparative study with magnetic resonance imaging, computed tomography and tomo-myelography. AB - To delimit the vertical extension of epidural tumors of the spinal canal, tomo myelography and magnetic resonance imaging (MRI) are the most accurate examinations, MRI appearing to us to be even superior to tomo-myelography. To evaluate precisely the bone extension of tumor growth as compared with the epidural lesions, computed tomography (CT) is more accurate than the other methods mentioned. The choice between these three techniques is of great importance as an aid in selection the best therapeutic method in each particular case. PMID- 2980468 TI - Magnetic resonance imaging compared with computed tomography and myelography in the diagnosis of spinal masses. AB - Magnetic resonance imaging (MRI) is superior to computed tomography (CT) and myelography in the demonstration and delineation of the extent of syringomyelia. In the detection of intramedullary tumors MRI is more sensitive than CT and myelography. MRI provides additional information on the sagittal and frontal planes regarding the extent of tumors. In the diagnosis of disc prolapse MRI seems to be as accurate as CT or myelography. Additional information is available with MRI in the diagnosis of degenerated disc tissue. Spinal stenosis is easily recognizable. CT was superior in the differentiation of bony and disc protrusion. The results show that MR has opened up new possibilities in the diagnosis of spinal diseases and will lead to a reorientation of the diagnostic approach. PMID- 2980469 TI - Reciprocity of the neural growth in the Arnold-Chiari malformation. AB - It has been suggested (1) that nerves grow as plants grow. Extensive development of one nerve tends to retard its neighbours, lack of development tends to excite them to more active growth. This is a widespread neurobiologic phenomenon which may be traced up also in the pathomechanism of the Arnold-Chiari malformation: Impairment of the neural growth at the level of a lumbosacral meningomyelocele results in overgrowth of the cranio-cervical nerves associated with shortening of the cervical cord segments. A similar reciprocity, normal and pathologic, seems to work also between the nerve roots of the cauda equina and the lumbosacral segments of the spinal cord. PMID- 2980470 TI - Principles of the diagnostic system 'Orfevre' for computerized analysis in radiology. PMID- 2980471 TI - Magnetic resonance microscopy in neurologic models. AB - Magnetic resonance imaging techniques have been developed to permit imaging with slice thickness less than 1 mm and pixels of 50 x 50 microns. Special purpose gradient and radiofrequency coils and three-dimensional imaging techniques enable acquisition of images with sufficient signal to noise to utilize these microscopic picture elements. Live 200 g rats were imaged enabling clear definition of gray and white matter structures. Examples include the Sylvian aqueduct and the substantia nigra. Three-dimensional microscopic images of live chick embryos enabled definition of ventricles and brain parenchyma as well as measurement of T1 over the set of 16 contiguous 1.2 mm slices. PMID- 2980472 TI - Diagnostic pitfalls and artifacts in high field magnetic resonance imaging. AB - Similar to all other medical imaging modalities, magnetic resonance imaging (MRI) has various artifacts. Some artifacts are easily recognizable such as motion, while some are not very obvious and even sometimes mimic pathologic conditions. These artifacts, if not differentiated, could easily become diagnostic pitfalls for radiologists and lead to erroneous conclusions. We have studied some of the imaging artifacts associated with a 1.5 tesla MRI scanner and report some cases who could become potential diagnostic pitfalls. Causes for these artifacts, although some not proven, are also presented. PMID- 2980473 TI - Evaluation of high resolution digital subtraction angiography in neuroradiology. AB - A digital subtraction angiography (DSA) system has been modified for capabilities of a 1024 x 1024 matrix with 10 bit depth. Spatial resolution was increased with expansion of the matrix, but contrast resolution was not increased as determined by phantom experiments. Smaller focal spots and magnification factors were also important for improvement of spatial resolution. Clinical evaluation using carotid, vertebral, and cervical DSA with intra-arterial contrast injections demonstrated that there was slight improvement in the image quality, but the final diagnosis and management of patients were not changed with the use of high resolution DSA. Further improvements in image quality of DSA will be accomplished by improvements of image intensifiers and use of smaller focal spots. PMID- 2980474 TI - Automatic reregistration for correction of localized misregistration artifacts in digital subtraction angiography of the head and neck. AB - A new automatic computer program, to remove misregistration artifacts secondary to motions such as rotation, contraction and relaxation of the body, has been developed. This program is called automatic reregistration and consists of determination of distortion vectors of corresponding segments of the mask and live images by detection of the peak value of two-dimensional cross correlation, followed by obtaining distortion vectors of other pixels through interpolation. Shift of the coordinate of the distorted live image is subtracted. Clinical evaluation of this new program, in comparison with already developed remasking and manual reregistration, have proved that some misregistration artifacts, uncorrectable by the latter two methods, have been removed to some extent. The new program has proven to be clinically useful. PMID- 2980475 TI - Fifty-five thousand cerebral angiographies. AB - A large series of patients submitted to cerebral angiography at the Bellaria Hospital in Bologna are presented in a preliminary report. Experience gained is discussed from the standpoints of success versus failure and of the complications. It is concluded that direct percutaneous cerebral angiography, if used selectively, still has a role to fill in a modern neuroradiology department, because of its safety. It is especially well suited for the examination of older patients. For angiography of the external carotid and the vertebral artery femoral catheterization is more suitable. PMID- 2980476 TI - Clinical events following neuroangiography. A prospective study. AB - The authors prospectively evaluated the clinical event rate over a 72 hour period following femorocerebral angiography in 1002 procedures. The neurologic event rate was 1.3 per cent in the first 24 hours (0.1% permanent) and 1.8 per cent between 24 and 72 hours (0.3% permanent). The non-neurologic event rate (mostly hematomas) was 7.2 per cent. There were no deaths. One hundred and nine pieces of data per procedure were analysed statistically. Delayed cerebral ischemia does occur and could be secondary to the procedure. The low risk of judiciously performed cerebral angiography was confirmed. Carefully cataloguing all events for up to 72 hours neither verifies nor excludes angiography as the cause of deterioration. PMID- 2980477 TI - Simultaneous isotope and CT cisternography in the diagnosis and evaluation of cerebral fluid rhinorrhea. AB - A prospective study was carried out in 29 patients to test the validity of the combined and simultaneous use of isotope cisternography and CT cisternography in the diagnosis of cerebrospinal fluid rhinorrhea. One of the conclusions reached was that a combination of the two procedures possibly gives the easiest and most effective diagnosis. PMID- 2980478 TI - Investigation of the cerebrospinal fluid circulation by CT contrast cisternography in patients with psychiatric diseases. AB - In 52 out of 4,400 patients with psychiatric diagnoses referred for a computed tomography (CT) examination a communicating hydrocephalus was suggested on the basis of the conventional CT scan. These 52 cases were investigated with CT contrast cisternography. A disturbed cerebrospinal fluid circulation leading to a reflux of contrast medium into the lateral ventricles, visible for up to 24 hours, and no uptake of contrast medium in the sulci of the convexity of the hemispheres, was detected in 45 of the 52 patients. The clinical effects of shunting in 26 cases are demonstrated. The value of cerebral blood flow studies in such cases is also discussed. PMID- 2980479 TI - Gated magnetic resonance imaging of the intracranial cerebrospinal fluid spaces. AB - The pulsatile nature CSF flow in the cerebral aqueduct has been demonstrated by du Boulay using air cineventriculography, a technique which disturbs normal CSF dynamics. To investigate this phenomenon non-invasively, we studied 35 normal volunteers using high-resolution, cardiac-gated magnetic resonance imaging (MRI). Specifically, we wished to document changes in size, configuration and signal intensity of the CSF spaces as they related to time in the cardiac cycle. Results show that changes in size and configuration were measurable in the third ventricle only (size increased during systole in 7 of the 35 volunteers). Except for the lateral ventricles, some loss in signal intensity was seen in all CSF spaces at least during systole, in all 35 volunteers-findings consistent with du Boulay's. However, contrary to du Boulay's observations, asymmetric loss of signal, consistent with pulsatile CSF flow, was demonstrated in the foramen of Monro in 15 of the 35 volunteers. Based on the pattern of signal void at the level of the foramen of Monro and on the expansion of the third ventricle during systole we propose a theory of synchronous CSF flow at the foramen of Monro and aqueduct, which unifies our MRI findings with du Boulay's cineventriculographic observations. PMID- 2980480 TI - Changes in cerebral ventricular dimensions related to the cardiac cycle as seen with gated magnetic resonance imaging. AB - The dynamic changes in ventricular dimensions related to the cardiac cycle were evaluated by analyzing gated magnetic resonance images in normal volunteers and in several patients with ventriculomegaly. Small variations in dimensions of the lateral and third ventricles were observed shortly after cardiac systole. PMID- 2980481 TI - Mapping of normal and abnormal cerebrospinal fluid flow/motion patterns using steady state free precession imaging. AB - In a comparative study more than 35 brain magnetic resonance imaging examinations were analyzed by mapping the CSF flow/motion pattern using the steady state free precession technique with two different flow sensitivity directions. Significant deviations from the normal pattern were seen in ventricular enlargements due to obstruction with no evidence of CSF flow/motion and in normal pressure hydrocephalus with complex flow pattern in lateral ventricles, suggesting a diagnostic potential for this fast imaging technique with sensitivity to very slow flow. PMID- 2980482 TI - Volumes of pituitary adenomas related to hormone production, duration of symptoms and postoperative outcome. AB - In the period from December 1978 to February 1986, 292 patients with pituitary adenomas were operated transsphenoidally. We measured tumor volumes from CT scans and pneumoencephalograms. Tumor volumes varied from 0.5 to 160 cm3. The duration of the clinical history varied from 6 months to 24 years. All pituitary hormones were recorded pre- and postoperatively in each patient. We found a positive correlation between tumor size and duration of clinical symptoms, both in hormone producing (active) and non-producing (inactive) adenomas. Among the prolactin and growth-hormone producing adenomas there was also positive correlation between tumor size and hormone levels: small and medium sized hormone producing adenomas (volume 6 cm3 or less), showed normalization of the preoperatively increased hormone levels after surgery. In patients with hormone producing pituitary adenomas larger than 6 cm3, we still found elevated serum prolactin or serum growth-hormone levels postoperatively. This indicates that the largest tumors could not be completely removed surgically. Pituitary tumors should be operated before they reach a volume of 6 cm3. If non-surgical treatment is initiated, the patient has to be closely followed with computed tomography. PMID- 2980484 TI - Radiologic diagnosis of lateral parasellar masses and their classifications. AB - Computed tomography (CT) and cerebral angiography of lateral parasellar masses were reviewed and classified into 6 categories according mainly to operative findings, as: 1) intradural or dural masses, 2) pituitary fossa masses, 3) intracranial extradural masses, 4) bony or cartilaginous masses, 5) masses of sphenoid sinus origin, and 6) carcinomas. Each of the 6 categories shows characteristic combinations of 3 cardinal radiologic features. These are: a) transverse displacement of the ganglial and cavernous carotid arteries on antero posterior carotid angiography, b) erosion or destruction of bone in the lateral parasellar region, and c) sphenoid sinus involvement on CT and conventional radiography. Use of this classification scheme based on the 3 cardinal radiologic features described should be of considerable aid in the diagnosis of lateral parasellar masses. PMID- 2980483 TI - Magnetic resonance imaging of tumours of the sellar region. Evaluation of treatment with bromocriptine retard. AB - Magnetic resonance imaging (MRI) with a superconductive 0.5 T magnet has been used in 10 patients with pituitary adenomas to evaluate the effect of bromocriptine treatment. Seven patients had prolactinomas, one a growth hormone (GH) producing adenoma and two patients null cell adenomas with slight hyperprolactinaemia. A single dose of 50 mg bromocriptine was given intramuscularly in a slow release preparation, followed after 6 weeks by 5 mg/day orally. Significant reduction of serum prolactin was noticed in all patients. Visual field defects disappeared rapidly. In all prolactinoma patients and in the patient with the GH producing adenoma MRI showed a decrease in tumour size. In 5 of these 8 patients MR signal analysis, aided by the use of a subtraction technique, clearly visualized the development of large intratumour cysts. In one further patient with a null cell adenoma there was also a rapid development of an intratumour cyst and an increase in tumour size. This patient was treated with transsphenoidal needle aspiration. In all tumour cases an increase in intensity quotients based on T1 and T2 relaxation times was calculated. PMID- 2980485 TI - Magnetic resonance imaging of the opto-chiasmatic region. Report on 276 cases. AB - A total of 276 lesions in the sellar and parasellar region were studied with magnetic resonance imaging using two different magnets of 0.15 and 0.5 T, respectively. Examination in the sagittal plane is recommended. T1 weighted images will give anatomic details, T2 weighted images the tissue characteristics. No marked differences with regard to diagnostic results were noted with the two different magnets, although the signals appeared to be more intense when using a 0.5 T magnet. PMID- 2980486 TI - Nasal dermal sinuses and cysts. AB - Nasal dermal sinuses and cysts are best detected by careful clinical inspection of the face for tiny ostia or widening of the nasal bridge and by detailed radiologic evaluation of the nasal bones, nasal septum, crista galli and falx. The sinus tracts may remain superficial to bone, extend partway into the nasal septum or pass far posterosuperiorly beneath the nasal bones, through the foramen cecum, and anterior to or through a bifid crista galli to lie between the leaves of the anterior falx. Glabellar sinuses may pass through the midline frontonasal suture to the foramen cecum, crista galli and falx. These courses correspond to embryonic midline diverticula of dura (with or without arachnoid and brain tissue) that protrude into a) the fonticulus nasofrontalis and b) the prenasal space situated behind the nasal bones and in front of the nasal capsule, occasionally reaching the surface ectoderm. Persistence of variable portions of these diverticula results in the clinical spectrum of nasal sinus tracts, fibrous cords, interposed (epi)dermoids, meningoceles, encephaloceles and 'gliomas'. PMID- 2980487 TI - Magnetic resonance imaging of the nasopharynx and skull base. AB - Ten patients with a normal nasopharynx and 36 patients with pathology of the nasopharynx or skull base were studied with magnetic resonance imaging (MRI). Four millimeter thick axial, coronal, and often sagittal scans were obtained using SE 500/28 and SE 2000/56 pulse sequences. The normal scans were compared with cadaver cryosections. Correlation was made with computed tomography (CT) scans as well as with clinical history and physical examination whenever possible. MRI clearly provided superior information compared with CT and other imaging studies. Advantages of MRI over CT include 1) the delineation of tumor infiltrations of the longus colli muscles, rectus capitus muscles, and muscles of mastication, 2) demonstration of tumor extensions into the skull base or parapharyngeal spaces, and 3) demonstration of both normal and pathologic cranial nerves. For all these reasons, MRI is currently the imaging study of choice for pathology of the nasopharynx and skull base. PMID- 2980488 TI - Magnetic resonance imaging and computed tomography of choroidal detachment. AB - The magnetic resonance imaging (MRI) and computed tomography (CT) findings in 9 patients with hemorrhagic choroidal detachment, serous choroidal detachment and/or ocular hypotony, choroidal effusion and 6 patients with exudative retinal detachment are described. Subacute hemorrhagic choroidal detachment appeared as moderately hyperintense image in T1 weighted MR scans and as hypointense in T2 weighted scans. Chronic hemorrhagic choroidal detachment was seen as hyperintense image in T1 and T2 weighted scans. Serous choroidal detachment and choroidal effusion appeared as moderately hyperintense image in T1 and T2 weighted scans. The exudative retinal detachment characteristically appeared on MR scans as a dependent hyperintense object in T1 and T2 weighted scans. PMID- 2980489 TI - Papilledema. Neuroradiologic evaluation of optic disc protrusion. AB - Patients presenting with elevated intracranial pressure (ICP) and papilledema in reality harbor a large degree of simple optic disc protrusion. Evaluations with dynamic CT demonstrate that there is depressed perfusion of the optic papilla in severe increased ICP in subjects with papilledema/protrusion as opposed to normal patients. This supports the theory that increased ICP leads to blindness by the mechanism of an increased pressure gradient at the optic nerve head resulting in ischemia and irreversible optic nerve atrophy. PMID- 2980490 TI - Computed tomography of lymphoproliferative disease of the orbit. Report of 50 patients. AB - Orbital lymphoproliferative disease is one of the most common causes of exophthalmos and palpable masses in the anterior part of the orbit. Axial and coronal computed tomography (CT) is the method of choice for localizing the lesion and determining the extent of involvement. The CT features are non specific but are highly suggestive in the proper clinical setting. The most important CT findings consist of: round, oval or elongated lobular masses, commonly in the extraconal space, with intraconal extension in large tumors. In some lesions, linear infiltrates radiate from the bulk of the mass into the adjacent fascial compartments. Tumors in proximity to the globe, wrap around the scleral-uveal coat causing no indentation but displacement of the globe from the mass effect. Similar molding occurs along the orbital walls, especially laterally from extraconal lesions. Bone destruction or remodeling of bone is rarely seen in orbital lymphoma. No diagnostic enhancement patterns were observed following contrast infusion. The majority of lymphoid tumors were situated anteriorly and superiorly. PMID- 2980491 TI - Combined magnetic resonance imaging and spectroscopy in experimental regional injury of the brain. Ischemia and impact trauma. AB - Combined magnetic resonance imaging (MRI) and spectroscopy (MRS) allows unique experimental insights into the central nervous system (CNS) disorders. Clinical applications are forthcoming. In order to address the potential clinical uses of MRI/MRS, experimentally induced focal brain lesions were evaluated with this modality. Focal lesions more closely mirror clinical situations than do global insults, but have rarely been the focus of MRS studies. Fluid-percussion trauma in 48 rats, focal ischemia in 18 rats and in 16 mongrel dogs were produced, with various degrees of severity. A discrepancy between temporal evolution data of MRI and MRS was found: MRI always showed an increase in lesion extension over time while 31P and 1H MRS almost always showed improvement. The severity and evolution of these MRS findings was surprising, and differed from the results reported for global brain injuries. Besides possibly reflecting real improvement in the metabolic state, other explanations for the phenomena exist. Diffusion of inorganic phosphate out of the regional site of injury and its reincorporation for more prominent lactate build-up in regional injury are possible. Therefore the use of MRS to predict metabolic and clinical outcomes of regional brain injury requires methodologic caution, but its combination to MRI offers an unprecedented tool for studies of focal CNS pathology. PMID- 2980492 TI - Young stroke patients. An angiographic study. AB - In 59 young stroke patients without predisposing conditions (thromboembolic mechanisms, diabetes, cardiac disease, coagulation disturbances), the angiographic findings and the clinical observations in combination make arterial dissection the most probable diagnosis in 45 patients. In an additional 7, this diagnosis remains as a possible diagnosis. It is suggest that in patients under 40 years of age with no predisposing disease and with sudden onset of neurologic deficits, preceded or accompanied by headache and/or neck pain and often associated with physical strain, arterial dissection should be regarded as the most likely diagnosis. PMID- 2980493 TI - Magnetic resonance imaging of brain tissue displacement and brain tissue water contents during progressive brain compression. An experimental study in dogs. AB - Continuous recording of vital physiologic parameters and magnetic resonance imaging (MRI) was performed during continuous expansion of extradural rubber balloons in spontaneously ventilated anesthetized dogs. Initial physiologic changes appeared when the pontine cistern was compressed while respiratory arrest was regularly paralleled by marked tentorial and tonsillar herniation. Local increase in brain tissue water progressed during brain compression and was most marked in white matter of the brain. This was related to the regional decrease in cerebral perfusion pressure. The correlation between the morphologic and physiologic course of events during brain compression has a predictive value. PMID- 2980494 TI - Magnetic resonance imaging of water intoxication. An experimental study in dogs. AB - Sequential magnetic resonance imaging (MRI) was performed to correlate signs of herniation and increase in local brain tissue water content with continuous changes in vital physiologic parameters during progressive water intoxication in anaesthetized dogs. The intracranial pressure increase ultimately resulted in respiratory and cardiac arrest. MRI concomitantly showed an increase in local brain water content starting and dominating in the cerebral cortex but progressing to all parts of the central nervous system. The late appearance of transtentorial pressure gradients and of brain herniation suggests that development of cerebral edema occurs in at least two stages, an intracellular osmotic edema appearing first, being followed by an ischemic edema related to a progressive decrease in local perfusion pressure. PMID- 2980495 TI - Magnetic resonance imaging of cerebral compression and local brain tissue water content during continuous extradural bleeding. An experimental study in dogs. AB - Continuous recording of vital physiologic parameters and magnetic resonance imaging were performed during progressive extradural bleeding in spontaneously ventilated anesthetized dogs. The volume of the bleeding could exceed the intracranial volume by 500 per cent due to the formation of an extradural arteriovenous shunt. The rate of hematoma growth was dependent on the area of stripped dura. Midline shift, herniation and lateral ventricle compression occurred early, corresponding to changes in vital parameters. Local brain water increased non-linearly with white matter preceding gray matter. Water increase started first in the ipsilateral hemisphere followed by the contralateral hemisphere, brain stem and cerebellum. The correlation between local tissue water increase and local perfusion pressure supports the notion of an ischemic brain edema being formed during an extradural bleeding. PMID- 2980496 TI - Evaluation of intracranial pressure rebound after evacuation of intracranial expanding lesions. An experimental study in dogs. AB - Sequential magnetic resonance imaging was used to follow brain displacement, signs of herniation and increase in local brain tissue water content during expansion and after evacuation of an extradural balloon in anesthetized, artificially ventilated dogs. A fatal intracranial pressure (ICP) rebound occurred if the cerebral perfusion pressure (CPP) was critically reduced to 20 mm Hg for more than half an hour. Despite reduction of brain displacement after balloon evacuation brain water content continued to increase. Compression of CSF outflow pathways and signs of herniation remained. CPP continuously fell to zero. ICP rebound is a grave situation significantly influenced by a large increase in brain tissue water. PMID- 2980497 TI - Cephalic magnetic resonance imaging of boxers. Preliminary results. AB - A prospective study of 52 boxers, 20-60 years, 13 of them amateurs and 39 professionals, has been made using magnetic resonance imaging (MRI) at 0.15 T. Twelve of the boxers were still active, while the other 40 had retired after about 10 years of boxing. Multi-slices in two planes without gap were performed using T1 and T2 weighted sequences. The findings were correlated to the number of matches, the number of knock outs, the severity of head trauma, the style of the boxer, and to the clinical findings and the findings at EEG. There was a good correspondence between the presence of lesions and the style of the boxer. The changes noted--mainly atrophic--occurred among those with capacity to 'stand punishment'. MRI offers an opportunity to evaluate the effect of trauma and the information obtained may be used to prevent disabilities among boxers. PMID- 2980498 TI - Magnetic resonance imaging of traumatic sinus and mastoid bleeding. AB - Magnetic resonance imaging (MRI) characterized intrasinus hemorrhage in 8 out of 10 patients with computed tomography (CT) evidence of post-traumatic sinus opacification. Fractures of sinus walls were demonstrated as disruptions in the hypointense line of the bony wall, as displacement of this line, or by increased signal from blood or sinus effusion within the fracture. Seven patients with fractures of the temporal bone were more difficult to evaluate because of the hypointensity of air trapped within the fracture line mimicing the bony wall and that within the middle ear mimicing the ossicles; 1.5 tesla MR proved superior to CT in demonstrating associated extra- and intracerebral hemorrhage. PMID- 2980499 TI - Xenon/CT cerebral blood flow determination following cranial trauma. AB - Xenon/CT cerebral blood flow (CBF) analysis has been utilized in the evaluation of 35 patients sustaining severe cranial trauma. The patient population included closed head injury, focal hematoma, and the sequelae of trauma including coma, persistent alteration of consciousness, and brain death. Xe/CT CBF analysis has proven helpful in the determination of appropriate degrees of hyperventilation therapy for the head injured patient. The technique is helpful for determining both local and remote changes of CBF as an aid in deciding upon surgical intervention. Severely decreased flows, compatible with brain death, can be demonstrated. The technique shows promise in the evaluation of physiologic changes accompanying therapy for the injured brain. PMID- 2980500 TI - Low field magnetic resonance imaging and computed tomography in the demonstration of posttraumatic brain abnormalities. AB - Thirty patients with a history of head trauma and who had neurologic or neuropsychologic disturbances were examined both with computed tomography (CT) and magnetic resonance imaging (MRI) (0.02 T). The examinations were performed 2 months to 30 years after the trauma. Low field MRI seems to be more sensitive than CT in the demonstration of intraparenchymal post-traumatic abnormalities. In the evaluation of central or cortical brain atrophy MRI and CT were equivalent. Even some patients with the primary diagnosis of concussion had demonstrable organic brain lesions. PMID- 2980501 TI - The spectrum of multiple sclerosis lesions using a multiple spin echo pulse sequence and a high field strength. AB - Seventy-one patients having clinical laboratory findings consistent with multiple sclerosis (MS) were imaged with a 1.5 tesla MR instrument using multiple spin echo sequences (TR = 2,000 ms and TE = 30, 60, 90, 120 ms). Multiple spin echo is a sensitive method for detecting MS lesions. Sixty-seven patients (94%) demonstrated lesions consistent with MS. With the exception of those located in the cortex, optic nerve and chiasm the lesions detected correlated with pathologic data. Lesions of the cerebral hemispheres, corpus callosum and cerebellum were generally multiple while lesions of the brain stem and optic tracts were generally singular. The majority of the lesions were associated with the white matter tracts of the cerebral hemispheres and brain stem. PMID- 2980502 TI - Acute vertebral-basilar thrombosis. Angiologic-clinical comparison and therapeutic implications. AB - The clinical-angiologic data and the clinical outcome in 66 patients with acute vertebro-basilar thrombosis treated with antiplatelet agents, anticoagulants, or fibrinolytic agents are presented. Forty-three patients were treated with local intraarterial infusion of streptokinase or urokinase proximal to the thrombotic occlusion; 14 patients in this group (33%) survived, only one of whom was in deep coma when therapy was started. Twenty-three patients did not receive fibrinolytic therapy; 4 of these patients (17%) who exhibited mild brainstem-related symptoms survived. This study implies that early diagnosis and treatment in vertebrobasilar stroke in progression may achieve improved survival. PMID- 2980503 TI - Relaxation times of apparently normal cerebral white matter in multiple sclerosis. AB - The magnetic resonance imaging (MRI) characteristics of the apparently normal cerebral white matter were studied in a group of 35 patients with clinically definite multiple sclerosis (MS). The proton relaxation times were compared with those in normal control subjects and patients with clinically isolated neurologic lesions of a type common in MS. Significant differences were found between all the patient groups and the control subjects. Changes in the cerebral white matter may be an early feature of MS. PMID- 2980504 TI - Lesions of high signal intensity at magnetic resonance imaging in vascular dementia. AB - Twenty-five patients with vascular dementia, 18 patients with senile dementia of Alzheimer type (SDAT) and 17 normal persons were examined by magnetic resonance imaging. Lesions of high signal intensity were always recognizable and were frequently very marked in patients with vascular dementia, while they were absent or mild in patients with pure SDAT and in non-demented patients without vascular disease. The possible nature of this disease as well as the role played in determining the type of dementia are discussed. PMID- 2980505 TI - Periventricular high signal lesions and signal void on magnetic resonance imaging in hydrocephalus. Diagnostic and prognostic significance. AB - The presence of periventricular high signal lesions and ventricular signal void was tabulated and compared in three groups: patients with unshunted hydrocephalus (n = 24), cerebral atrophy (n = 14), and a subgroup of treated hydrocephalus subjects studied both pre- and post-shunting (n = 6). We found that T2 high signal lesions and ventricular signal void are common in both hydrocephalus and atrophy and are of no value in distinguishing between the two. The pre-treatment presence or absence of periventricular high signal lesions and signal void were of no use in predicting the clinical outcome of shunting in hydrocephalus. PMID- 2980506 TI - Hemispheric white matter lesions in the elderly prone to falling. AB - Brain computed tomography studies of two groups of elderly (mean age 83.3 years, SE 1.7) with no neurologic diagnoses were compared. Sixteen of the subjects were non-fallers and the remaining 17 had frequent unexplained falls. The group of fallers had a significantly greater degree of white-matter hypodensity of the elderly (Student's t-test = 2.7, p less than 0.01). PMID- 2980507 TI - Computed tomography in chronic alcoholism. AB - A series of 327 chronic alcoholics was studied by computed tomography (CT) and compared with 419 age-matched controls. The inner and outer CSF spaces of the brain were measured in the two groups. The values obtained were tested by a stepwise discriminant analysis. On this basis the following six parameters proved to be the best signs for distinguishing chronic alcoholics from non-alcoholics: the widths of the third ventricle, of the cistern of the quadrigeminal plate, of the interhemispheric fissure, and of the parietal and temporal sulci, the Huckman number, and the presence or absence of cerebellar atrophy. PMID- 2980508 TI - Cerebellar atrophy diagnosed by computed tomography and clinical data. AB - The diagnostic relevance of computed tomography (CT) in the classification of cerebellar atrophy or degeneration is unclear. Twenty-one patients with cerebellar atrophy at CT were studied and the findings were correlated to clinical data. Based upon such data the material was divided into two groups. In the first group (12 patients), with signs of cerebellar deficiency, 6 cases presented familial hereditary ataxia, and olivopontocerebellar atrophy of the Menzl type, 3 ataxia telangiectasia (Louis-Bar syndrome), 2 olivopontocerebellar atrophy of the sporadic type, and 1 adrenoleukodystrophy. In the second group (9 patients), without cerebellar deficit, cerebellar atrophy was found only occasionally. In all of them, there was cerebral atrophy. Clinically manifest cerebellar deficiency and cerebellar atrophy as evident at CT was mainly found in patients with familial genetic disorders. Cerebellar and/or vermian atrophy without clinical signs of cerebellar deficiency were observed only occasionally and were not specific. PMID- 2980509 TI - The role of digital subtraction angiography in neuroradiology. An evaluation of intravenous and intra-arterial digital subtraction angiography. AB - A few years after the introduction of intravenous digital subtraction angiography (DSA) in clinical practice it became clear that, although the method had been developed as an alternative to arterial angiography, especially for the analysis of cerebral ischemia, its application was not as successful as the earlier reports had indicated. Based on the results of a study comparing intravenous DSA, intra-arterial DSA, and the Doppler examination, in 168 consecutive patients, a diagnostic algorithm for the management of these patients is proposed. A second and more important application of digital subtraction is its use with arterial angiography, especially with a high resolution system configuration. PMID- 2980510 TI - The radiologic investigation of temporal lobe epilepsy. Value of high resolution CT with temporal orientated sections. AB - The value of high resolution computed tomography of the temporal lobes using temporal lobe orientated sections and intrathecal contrast is described. The results of this technique are correlated with the pathologic findings in 13 patients subsequently proven to have temporal lobe gliomas. These patients all presented with drug resistant temporal lobe epilepsy and underwent temporal lobectomy. This technique was successful in detecting the tumours in 11 of 13 patients. The average size of the tumours was just over 2 cm. The importance of detecting these small tumours is emphasised because of the profound effect this has on management. PMID- 2980511 TI - In vivo measurement of dopamine receptors in pituitary adenomas using positron emission tomography. AB - Patients with pituitary adenomas were examined with positron emission tomography (PET) with the administration of the 11C-labelled dopamine-D2 antagonists N methylspiperone and raclopride. The studies were repeated after protection of the D2-receptors with Haloperidol to enable a separation of specific and unspecific receptor binding. The receptor binding was evaluated by visual inspection and with the application of a kinetic model. The results showed marked specific dopamine-D2 receptor binding in the prolactinomas and minimal or no such binding in the hormonally inactive adenomas. The two tracers 11C-raclopride and 11C-N methylspiperone showed qualitatively the same result although raclopride resulted in a higher tumor to normal brain ratio. In conclusion, PET is a valuable complement to other radiologic techniques like computed tomography and magnetic resonance imaging in the evaluation of pituitary adenomas. An assessment of the dopamine-D2 receptors in the adenomas has a direct influence on the choice of treatment because adenomas with high amounts of receptors are in most cases effectively treated with dopamine agonists like bromocriptine. PMID- 2980512 TI - Amino acid metabolism in pituitary adenomas. AB - Patients with pituitary adenomas were examined with positron emission tomography with the administration of 11C-labelled methionine. In a few patients comparative studies using the two stereo-isomers L- and D-methionine were also performed. The results showed a rapid distribution of D-methionine within the tumor tissue without signs of irreversible trapping of the tracer. L-methionine on the other hand was rapidly irreversibly trapped indicating the metabolic fate. It is concluded that 11C-L-methionine can be used to quantify amino acid metabolism in the tumors. The amino acid metabolism was evaluated in a group of adenomas before and after treatment with bromocriptine. The results indicate a marked reduction of the amino acid metabolism in prolactinomas amounting to 60%. These tumors at a later time responded with tumor shrinkage. Hormonally inactive tumors showed no effect of treatment. PMID- 2980513 TI - Relation between tissue nature and (18F) fluorodeoxyglucose kinetics evaluated by dynamic positron emission tomography in human brain tumors. AB - Using dynamic positron emission tomographic (PET) scans, kinetic rate constants in the (18F) fluorodeoxyglucose (18FDG) model and glucose consumption rate were measured in human brain tumors. In the tumor tissue, all rate constants were higher on the average in the glioma grade IV and the non-glial tumor groups. The k*3 showed wide deviation in the glioma grade IV group. Discrepancy was found between the k*3 and the glucose consumption rate in several cases. The k*1 correlated with the neovascularity in tumors confirmed at angiography. Analysis of the rate constants might be useful for interpreting tumor nature and predicting its malignancy. PMID- 2980514 TI - Single photon emission computed tomography of brain tumors with 201TIC1. AB - Single photon emission computed tomography (SPECT) with 201TIC1 was performed in 27 patients with brain tumors. Findings on SPECT were compared with those on planar images, computed tomography (CT) and, in 4 cases, magnetic resonance imaging (MRI). At SPECT the tumors were shown as areas with higher accumulation of the tracers than the surrounding brain tissue in 24 cases and a better discrimination was obtained than in planar images. Deep-seated lesions were especially visualized. In some cases SPECT defined lesions that were not clearly delineated on CT or MRI. SPECT with 201TlCl can be of clinical use in the diagnosis of brain tumors. PMID- 2980515 TI - Cerebral glucose metabolic change after high-dose methotrexate treatment in patients with acute lymphocytic leukemia. AB - An investigation was carried out to study the changes in the cerebral metabolic rate for glucose (CMRGlc) after high-dose methotrexate infusion (HD-MTX) in leukemic children, compared with the rate before such treatment. This is the first report on the relation between HD-MTX and the CMRGlc. An infusion of HD-MTX significantly reduced the glucose metabolism. It is suggested that this depression of the CMRGlc may have been associated with acute neurotoxicity of MTX. Measurement of the CMRGlc may be of value in detecting neurotoxicity of MTX. The reasons for these metabolic alterations need further investigation. PMID- 2980516 TI - Positron emission tomography in patients with a primary intracerebral hematoma. AB - Using positron emission tomography, the regional cerebral blood flow (CBF), oxygen extraction fraction (OEF) and oxygen consumption (CMRO2) were studied in 21 patients with hypertensive intracerebral hematomas. The study showed a very localized decrease of CBF and CMRO2 around a hematoma, and revealed in two cases (9.1%) distinct luxury perfusion around it. There was diffuse reduction of CBF with increased OEF in hematomas larger than 4.5 cm in diameter. The results suggested that surgical evacuation should be considered for hematomas with a diameter exceeding 4.5 cm. PMID- 2980517 TI - Regional cerebral blood flow and metabolism in patients with chronic obstructive carotid disease. AB - Regional cerebral blood flow (CBF), oxygen extraction fraction (OEF) and oxygen consumption (CMRO2) were studied by PET in 26 patients with chronic occlusion(s) or severe stenosis of the internal carotid or middle cerebral artery(ies) without a large (greater than 2 cm) infarction. Most of them showed a coupled decrease of CBF and CMRO2 in the internal carotid distribution. In 5 patients (19%), the CBF in the least perfused but structurally normal tissue was reduced to critically low values while maintaining CMRO2, considerably increasing OEF. Since detection of a lesion with hemodynamic insufficiency was important for the indication for an EC/IC bypass operation, only 19 per cent of the present series would have had an indication for surgery. PMID- 2980518 TI - Edema and atrophy following cerebral stroke. A prospective and consecutive study. AB - Ninety-six consecutive stroke patients were examined by sequential CT scans. Edema in the acute/subacute state and atrophy in the chronic state were evaluated by measuring a frontal horn index, septum-caudate distance and width of cortical sulci. In the acute state the degree of edema increased with increasing size of the infarct. Cerebral swelling was more marked and persisted for a longer period in patients having intracerebral hematomas than in patients with ischemic infarcts. Surprisingly, edema was present in the unaffected hemisphere as well. Development of atrophy was seen in the affected hemisphere in patients having large infarcts and intracerebral hematomas. Atrophy did not develop in the unaffected hemisphere. PMID- 2980519 TI - Assessment of regional cerebral blood flow with 123I amphetamine single photon emission computed tomography in cerebrovascular disease. Semiquantitative analysis comparing IMP SPECT with the 133Xe inhalation method. AB - To compare the capacity of three-dimensional IMP single photon emission computed tomography (SPECT) with that of the two-dimensional 133Xe method in detecting focal perfusion abnormalities in cerebrovascular disease, flow measurements were performed with both methods in twenty patients. In order to enable correlation with 133Xe regional cerebral blood flow (rCBF) IMP SPECT images were subdivided into areas extending from the outer to the inner part corresponding to 32 regions of the 133Xe Cerebrograph. Between IMP counts/pixel and 133Xe rCBF, inter hemispheric ratios correlated better in severe ischemia. Absolute mean values correlated in the affected hemisphere but not in the unaffected hemisphere. Regional evaluations revealed tendencies of IMP SPECT to be sensitive for low flow areas but to underestimate high flow areas, i. e. the 'luxury perfusion syndrome', as compared with the 133Xe method. PMID- 2980520 TI - Positron emission tomography of hydrocephalus. Metabolic effects of shunt procedures. AB - Five patients with hydrocephalus were studied with carbon-11-2-deoxyglucose or 2 deoxy-2-(18F) fluoro-D-glucose and positron emission tomography both prior to and following ventricular shunting. Four subjects had communicating hydrocephalus; the fifth had aqueductal stenosis, two patients had hydrocephalus for three months or less. The three chronic patients were felt to have hydrocephalus for three years or more. After shunting ventricular size decreased in all patients, and all patients showed clinical improvement. The glucose cerebral metabolic rates increased after shunt in the two subjects with recent onset hydrocephalus but paradoxially decreased in the three chronic patients despite clinical improvement. These findings suggest that the cerebrum was metabolically hyperactive prior to shunt due to an unknown mechanism and presumably in response to the presence of hydrocephalus. A dissociation may also exist in the post-shunt period between cerebral metabolism and cerebral blood flow. PMID- 2980521 TI - Regional cerebral blood flow changes in aging. AB - Regional cerebral blood flow (rCBF) changes during the aging process were studied, using 131Xe inhalation and emission computed tomography in 38 healthy, non-hypertensive, alert and active volunteers (23-65 years of age). The rCBF showed a slight but statistically non-significant negative correlation. As regards cerebrovascular resistance (CVR), significant correlation to aging was observed, and there was an increase of CVR after the forties compared with the twenties. This appeared to be due to the positive correlation of blood pressure to aging. PMID- 2980522 TI - Regional cerebral glucose metabolism in anxiety disorders studied with positron emission tomography before and after psychosurgical intervention. A preliminary report. AB - Regional cerebral metabolic rate of glucose (rCMRGl) was studied with positron emission tomography in patients suffering from severe anxiety disorders undergoing capsulotomy, an anti-anxiety psychosurgical intervention in which fronto-limbic connections are interrupted. Preliminary observations on five patients show a statistically significant reduction in rCMRGl postoperatively in the orbitomedial frontal cortex, a region knonw to be of relevance for the capsulotomy effects. Clinically, 4 of 5 patients improved after surgery. The study may provide new information on the pathophysiology of anxiety. PMID- 2980524 TI - Regional cerebral blood flow determined with single photon emission computed tomography and positron emission tomography. A comparative study. AB - The regional uptake of 99Tcm-labelled HMPAO has been measured with single photon emission computed tomography (SPECT). The potential value of 99Tcm-HMPAO as a flow tracer was evaluated. The uptake of 99Tcm-HMPAO as a function of time indicated that the tracer was trapped in the brain. The uptake images were compared with results from quantitative regional cerebral blood flow (rCBF) measurements using positron emission tomography (PET) and with 11C-fluoromethane used as a tracer. There was a strong correlation between the uptake of 99Tcm HMPAO and rCBF. The conclusion was that the uptake of 99Tcm-HMPAO was dependent on rCBF. PMID- 2980523 TI - Applications of a computerized adjustable brain atlas in positron emission tomography. AB - A computerized brain atlas, adjustable to the patient's anatomy, has been developed. It is primarily intended for use in positron emission tomography (PET), but may also be employed in other fields utilizing neuro-imaging, such as stereotactic surgery. The atlas is based on anatomic information obtained from digitized cryosectioned cadaver brains. It can be adjusted to fit a wide range of individual brains with reasonable accuracy. The corresponding transformation is chosen so that the modified atlas agrees with a set of CT or MR images of the patient. The computerized atlas can be used to facilitate and improve the quantification and evaluation of PET data by: enabling the merging and comparison of results from different individuals or groups of individuals; serving as a vehicle in the comparison of different examinations of the same patient, thus reducing the need of reproducible fixation systems; supplying external information to be used in the image reconstruction, such as proper three dimensional regions of interest; improving the attenuation and scatter corrections; helping to select suitable patient orientation during the PET study. By applying the inverse atlas transformation to the PET data volume it is possible to relate the PET information to the anatomy of the reference atlas. Reformatted PET data from different patients can thus be averaged, and averages from different categories of patients can be compared. The method will facilitate the identification of statistically significant differences in the PET information from different groups of patients. PMID- 2980525 TI - Evaluation of a 99Tcm bound brain scanning agent for single photon emission computed tomography. AB - D,L HM-PAO-99Tcm (PAO) is a lipophilic tracer complex which is avidly taken up by the brain. We have compared the regional distribution of PAO with regional cerebral blood flow (CBF). CBF was measured by single photon emission computed tomography (SPECT) by Tomomatic 64 after 133Xe inhalation in 41 patients. With the same SPECT the distribution of PAO was measured after intravenous injection. High resolution (HR) and low resolution (LR) studies were performed yielding a resolution of 6-10 mm (HR) and 15-20 mm (LR). PAO images showed close resemblance to 133Xe CBF tomograms. Only 20 per cent of the (decay corrected) brain counts were lost during 24 hours. PMID- 2980526 TI - Magnetic resonance imaging of hemorrhagic cerebral infarction. AB - A comparative study between magnetic resonance (MR) and computed tomography (CT) showed MR to be more sensitive than CT for detecting subacute and chronic hemorrhage into cerebral infarction. The hemorrhage appeared as either high signal on T1 weighted images or low signal on T2 weighted images, depending on the age of the hemorrhage. Cortical hemorrhagic infarction showed a characteristic gyral pattern. Methemoglobin and hemosiderin served as markers of previous hemorrhage into cerebral infarcts. PMID- 2980527 TI - 123I-N-isopropyl-amphetamine single photon emission computed tomography as a brain imaging technique in dementia. AB - Because of its binding to specific cerebral amine receptor sites 123I-N-isopropyl amphetamine (IMP) tracer activity can be used as a measure for the regional cerebral blood flow (rCBF) in the human brain. In 21 psychiatric in-patients an IMP perfusion study was performed using a rotating gamma camera system. The findings at single photon emission computed tomography (SPECT) were compared with different clinical parameters including conventional transmission computed tomography. In SPECT, two different patterns of decrease in IMP uptake could be identified. In patients with a history of cerebrovascular disease SPECT showed asymmetrical, multifocal microcirculatory defects without preference for either hemisphere. In patients with suggested Alzheimer type dementia the perfusion deficits involved the gray and white matter of the parieto-occipital lobes in a bilaterally symmetrical mode with variable extension. Thus, by using IMP-SPECT, it was possible to define the underlying pathologic condition in dementia. PMID- 2980528 TI - Minisymposium on technical aspects of magnetic resonance imaging. PMID- 2980529 TI - Magnetic resonance image synthesis. Clinical implementation. AB - Magnetic resonance (MR) image synthesis is a technique enabling the retrospective optimization of scanning parameters. This paper describes the methods used to enable the clinical implementation of this technique. It is shown that effective synthetic images can be generated from only three acquired images per slice. Data for 16 slices and TR times of 500 and 2000 ms can be efficiently acquired in a multi-slice multi-echo dual-TR pulse sequence. Computation of T1, T2, and density images and the subsequent synthesis of images for arbitrary TR and TE times can be performed at high speed with dedicated hardware. The method is seen as one of standardizing acquisition protocols and thereby improving patient throughput. Specific clinical applications are discussed. PMID- 2980530 TI - Spin echo and inversion recovery sequences for gadolinium-DTPA enhanced magnetic resonance imaging of intracranial tumors. AB - In 36 patients with intracranial tumors, gadolinium (Gd)-DTPA-enhanced SE 400/35, SE 800/35 and IR 1500/400/35 sequences were intraindividually compared. Contrast enhancement usually was shown to better advantage with the IR sequence. Post contrast SI of tumor tissue was isointense with white matter on IR images in many cases whereas SE sequences displayed enhancing tumor with high contrast versus brain. Post-contrast tumor delineation, therefore, was better with the SE sequences especially in white matter lesions without perifocal edema. The SE 400/35 sequence which allows for contrast-enhanced MRI within a relatively short time is suggested for Gd-DTPA studies of brain tumors. PMID- 2980531 TI - High accuracy and contrast resolution in ultra-low field magnetic resonance imaging. AB - Magnetic resonance imaging (MRI) at low magnetic field strengths is known to have a comparatively high contrast resolution. Current techniques increasing the signal to noise ratio 3 to 4 times allow for an improvement in the spatial resolution still maintaining the high contrast resolution. The contributing features to this advancement are improvements in software, development of new radiofrequency coils and image processing computors. The subsequent improvement in image evaluation and the access to added information make for a high accuracy in clinical diagnosis and research. The present potential of imaging at ultra-low (0.02 tesla) magnetic field strengths is illustrated with examples from clinical practice and research. PMID- 2980532 TI - Magnetic resonance imaging of the thoracical spinal cord and spine with surface coils. AB - We report the results obtained at 0.6 T with the use of surface coils in 77 patients who presented with obvious or suspected thoracic myelopathy. There were 16 patients with primary tumors diagnosed for the first time, 9 patients with residual tumors, 17 with metastatic neoplasms, 13 with hydrosyringomyelia, 5 with vertebral fracture and secondary pathology involving the cord, 5 with thoracic disc protrusions, 2 with multiple sclerosis, 2 with arteriovenous malformations, 2 with cystic arachnoiditis, and 6 with a variety of congenital abnormalities (low tethered cord, intradural lipoma, lipomyelomeningocele). The significantly improved signal to noise ratios provided by surface coils allow high quality thin section imagning which, combined with the high tissue contrast resolution obtainable with suitable pulse sequences, makes MR the preferred initial imaging technique over myelography and CT myelography. In an interesting spectrum of lesions, MR can obviate the need for traditional invasive procedures. PMID- 2980533 TI - Mathematical morphology for automatic detection of brain lesions at magnetic resonance imaging. AB - An original method of image analysis has been developed, using 'mathematical morphology', in order to detect brain tumors at magnetic resonance imaging of the head following injection of gadolinium-DPTA. The main steps of the analysis are as follows: 1) Spatial reduction by automatic determination of a region of interest, 2) elimination of noise by morphologic filtering (increasing alternate closing-opening sequence), 3) localization of tumor by searching for r-h maxima, and 4) extraction of tumor contours using three-dimensional structuring elements. The preliminary results of this automatic and robust method encourage further studies of the potential of tissue analysis as an aid to tumor diagnosis. PMID- 2980534 TI - Subtraction in magnetic resonance imaging. AB - Subtraction of images recorded with multiple echo, spin echo (SE) techniques was performed in phantom and clinical studies of patients with pituitary adenomas and gliomas. The tissue differentiation was enhanced in the subtracted images. PMID- 2980535 TI - Method for quantification of low flow velocities by magnetic resonance phase imaging. AB - The aim of this study was to compare the influence of flow in the velocity range 0 to 25 mm/s on modulus, phase, real and imaginary images obtained with a standard magnetic resonance scanner (Siemens Magnetom, 0.5 T), and to develop a simple method for determination of flow velocities in vivo from this information. Using a flow phantom, the flow dependent magnetic resonance imaging (MRI) signal has been studied as a function of flow perpendicular to the image slice with non doped water (simulating moving cerebrospinal fluid) as well as with water doped with Mn2+ (simulating moving blood) for each of the four mentioned image types. The results show a marked flow dependence on all types of images studied. The variation of the signal with flow in the modulus images is relaxation-time dependent in the studied velocity range and it is non-monotone for non-doped water. In the phase images, however, the variations are monotone and not dependent on relaxation times. In modulus images the curve shape is relatively independent on flow direction, while phase images are clearly dependent on flow direction in the studied velocity range. The signal versus velocity curves for the real and imaginary images show resemblance to those for the modulus and the phase images, respectively. It is concluded that the phase information can be used to generate a signal versus velocity calibration curve, which can be used to quantify low flow velocities in vivo. PMID- 2980536 TI - Magnetic resonance imaging in the evaluation of nimodipine-treated acute experimental focal cerebral ischemia. AB - We evaluated the sensitivity of magnetic resonance (MR) imaging in documenting effects of nimodipine in experimental focal cerebral ischemia. Twenty-five Sprague Dawley rats underwent unilateral occlusion of the middle cerebral artery and were imaged at different intervals thereafter. Neuropathologic and neurologic data were correlated with MR imaging results. Compared with controls, nimodipine treated rats showed a significantly smaller infarct size (p less than 0.001), as documented by MR imaging and confirmed by neuropathologic evaluations. A less intense signal on the T2 weighted sequence was found in nimodipine-treated rats in basal ganglia (p less than 0.001) and cortex (p less than 0.05). MR imaging may afford unprecedented diagnostic sensitivity in assessing pharmacologic efficacy in cerebral ischemia. PMID- 2980537 TI - Cerebrospinal fluid flow studied with gated magnetic resonance imaging during the various parts of the cardiac cycle. AB - The pulsatile movement of cerebrospinal fluid (CSF) through the Sylvian aqueduct has been studied in two normal volunteers. A standard magnetic resonance scanner was used as well as a routine spin echo sequence. Series of ECG-gated axial images were obtained perpendicular to the long axis of the Sylvian aqueduct. Previously it has been demonstrated by flow phantom experiments that the phase information can be used to obtain a linear relation between phase and flow velocity. By multiplying the CSF flow velocity by the cross-sectional area of the aqueduct of Sylvius obtained in each image, the CSF flow variation during the cardiac cycle could be demonstrated and measured. PMID- 2980538 TI - Quantitative analysis of intervertebral disc structure. AB - A reduction in signal intensity from the nucleus can be a feature of intervertebral disc disease. It has been established that T1 and T2 relaxation times of the nucleus decrease with age and that evidence of disc degeneration can be determined before classical clinical features are apparent. A robust multiple point algorithm has been developed which now enables proton density and T1 and T2 values to be computed for each pixel, providing images which are effectively spatial maps of these parameters. Such high resolution maps have provided quantitative data from volunteers, patients and cadavers. These studies have been carried out with specially constructed coils and the cadaveric information compared with cut sections. The spatial maps revealed detailed anatomic structures including the laminae of the annulus and relative levels of hydration. These levels, which are known to be related to the ability of the disc to withstand compressive loads, can now be measured in vivo. Analysis of the proton density and relaxation times in vivo has demonstrated that both water content and the chemical environment in the nucleus change during aging. The results are consistent with changes in the glycosaminoglycan content and fixed charged density measured by other chemical and physical techniques. Such detailed methods can be used to investigate the effects of aging and disease on disc structure and have enabled observations to be made of the effects of stress on the normal disc. PMID- 2980539 TI - Measurement of NMR relaxation times using the minimum number of scans. AB - NMR relaxation time measurements derived from a set of three multi-slice sequences (which can examine the whole brain in 30 min) have been investigated. T1 and T2 values are 5% and 20% too short, respectively. Noise in the measurements was less than 2%. Temporal instability was negligible. Spatial non uniformity was 1% (T1) and 3% (T2) within a single slice but 4% (T1) and 14% (T2) from slice to slice. Variations in cerebral tissue measurements have also been investigated: T1 varies by 6% and T2 by 12% over a population of normal controls. There can be significant patient-patient and topographical variations. PMID- 2980540 TI - Anesthetics change tissue proton NMR relaxation. AB - Previous studies have shown the treatment of experimental animals with therapeutic doses of pharmaceuticals in acute and chronic regimens has resulted in extensive (up to 33%) change in tissue proton NMR relaxation times. In this study, the effects of injectable anesthetics (Innovar, pentobarbital, ketamine and alpha-chloralose) were compared with inhalational anesthetics (halothane, enflurane and isoflurane). Proton NMR T1 and T2 relaxation times were obtained from excised tissues of anesthetized rats (6 each treatment). Sodium pentobarbital and alpha-chloralose showed no alterations, whereas Innovar produced increases in brain and kidney relaxation times and ketamine showed increases in the heart. More widespread and extensive changes were observed after 1.5 hours of inhalation anesthesia: liver, muscle and fat showed relaxation time decrease; heart, spleen and lung were increased. No change in water content, as measured by wet/dry ratios, was observed for any tissue. Therefore, intravenous anesthesia may be preferred to inhalational anesthesia for magnetic resonance imaging based on potential changes in tissue relaxation properties. In addition, the absence of a measurable effect on brain suggests that the mechanism for anesthesia does not involve major changes in brain water. PMID- 2980541 TI - The NMR signal decay characteristics of cerebral oedema. AB - Quantitative magnetic resonance (MR) imaging has been used to characterize vasogenic and triethyltin-induced cytotoxic cerebral oedema in cats, and the findings have been compared with the ultrastructural features of the lesions. Both normal and oedematous white matter yield non-linear T2 signal decay curves. The derived biexponential functions comprised a short T2 component representing intracellular water, and a long T2 component representing oedema fluid whose T2 was increased by an increase in free water of these tissue compartments. The relative sizes of the long T2 components were compatible with the sizes of the oedema spaces as determined ultrastructurally. Quantitative MR imaging can provide information which reflects the size and relaxation behaviour of the major tissue water compartments. PMID- 2980542 TI - Combined magnetic resonance imaging and bihemispheric magnetic resonance spectroscopy in acute experimental focal cerebral ischemia. AB - Combined magnetic resonance imaging (MRI) and 31P-magnetic resonance spectroscopy (MRS) were employed to evaluate acute regional cerebral ischemia in cats following middle cerebral artery occlusion (MCAO). MRI changes were first reliable 3 hours after MCAO; progressive increases in T1 and T2 relaxation times were noted for 12 hours after MCAO. Immediately after MCAO, 31P-MRS revealed decreased phosphocreatine (PCr) and increased inorganic phosphate (Pi); the PCr/Pi ratio fell from 2.23 +/- 0.40 to 0.68 +/- 0.04 (p less than 0.005). This relative decrease in high energy phosphates improved significantly by 3-5 hours after MCAO (0.67 +/- 0.04 versus 1.00 +/- 0.03, p less than 0.001). By 12 hours, the ratio had increased by 27 per cent. The dissociation between the progressive MRI changes and the gradual improvement in 31P-MRS suggests a difference between the processes affecting proton distribution and phosphate metabolism in acute regional ischemia. PMID- 2980543 TI - Thermoregulatory responses to clinical magnetic resonance imaging of the head at 1.5 tesla. Lack of evidence for direct effects on the hypothalamus. AB - Warming temperature sensitive neurons in the hypothalamus will induce thermoregulatory heat dissipation. Application of radiofrequency (RF) radiation to the head during magnetic resonance imaging (MRI) could, conceivably, heat the brain, causing a generalized peripheral vasodilation and result in a paradoxical and unnecessary decrease in body temperature. To evaluate the thermoregulatory responses to the RF power deposition used during MRI, we measured body (sublingual pocket) and skin temperatures in 15 patients immediately before and after MRI scans of the head. Ear-skin blood flow was determined by laser-Doppler velocimetry to assess local vasomotor tone. A high-field (1.5 tesla/64 MHz) MRI device (General Electric Company) with a coil designed for head/brain imaging was used in this study. Ambient conditions were room temperature 20-24 degrees C and relative humidity 40-50%. The specific absorption rate averaged over the head ranged from 0.83 to 1.20 W/kg. There was a slight but statistically significant elevation in body temperature (36.5 +/- 0.5 to 36.7 +/- 0.4 degrees C). Skin temperatures of the ear (30.0 +/- 1.2 to 32.0 +/- 0.9 degrees C) and forehead (32.4 +/- 0.5 to 32.8 +/- 0.5 degrees C) increased significantly, while hand (29.9 +/- 1.4 to 29.8 +/- 2.1) skin temperature was unchanged. Ear-skin blood flow also increased a statistically significant amount (average change 36%). The data indicate that there was predominantly surface heating associated with MRI of the head which stimulated a local vasodilating response (i.e. significant increase in ear skin blood flow).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980544 TI - Thermal effects of high-field (1.5 tesla) magnetic resonance imaging of the spine. Clinical experience above a specific absorption rate of 0.4 W/kg. AB - Current safety guidelines recommend limiting the exposure to radiofrequency (RF) radiation used for clinical magnetic resonance imaging to a whole body average specific absorption rate (SAR) of 0.4 W/kg. Since it may be desirable to image with SARs that exceed this level during MRI of the spine, we evaluated the thermal responses associated with these procedures. Body and skin temperatures were determined in 25 patients immediately before and after MRI. Since the eye is particularly susceptible to thermal injury, corneal temperature was also measured. High-resolution thermography was performed on three subjects to evaluate the surface heating pattern and identify potential thermal 'hot spots'. A 1.5 tesla/64 MHz MRI system with quadrature transmission and reception was used iN this study. The whole body average specific absorption rate ranged from 0.5 to 1.3 W/kg. Ambient conditions were room temperature 20-24 degrees C and relative humidity between 40 and 50 per cent. There was a slight but statistically significant (p less than 0.01) increase in body temperature after MRI (36.5 +/- 0.4 to 36.7 +/- 0.4 degrees C). Temperatures of the hand (30.4 +/- 1.4 to 31.2 +/ 1.0 degrees C), positioning isocenter (32.1 +/- 0.6 to 32.9 +/- 0.5), and cornea (32.5 +/- 0.6 to 32.9 +/- 0.5 degrees C) also increased a statistically significant amount. Thermographic imaging revealed normal heating patterns with no surface 'hot spots'. We conclude that the temperature changes associated with MRI of the spine at the SARs we studied were well below known thresholds for adverse effects and do not appear to be harmful to patients. PMID- 2980545 TI - Cerebral arterial constriction due to contrast media. AB - Angiography, either with Iohexol or Hexabrix, was carried out on a series of baboons, each of which had both contrast media, but on separate occasions. A double blind trial showed that in the carotid and middle cerebral arteries: Hexabrix generally caused vasoconstriction by 1.5 min, but in spite of a subsequent injection at 6 min constriction had largely disappeared by 20 minutes. Iohexol was generally followed by less severe constriction, but it may persist for longer and even show an increase at 20 min. PMID- 2980546 TI - Iohexol versus meglumine-Ca-metrizoate in cerebral angiography. A randomized double-blind cross-over study. AB - The tolerance and suitability of iohexol in cerebral angiography was compared in a clinical trial with meglumine-Ca-metrizoate in 20 patients. A preference for iohexol after injection into the external carotid artery in a paired comparison was established in 84.2 per cent of the patients. The difference was also statistically significant (p less than 0.01) with regard to the intensity of discomfort estimated according to a four grade scale. The milder sensations occurred more frequently with iohexol (1.21), while moderate as well as severe reactions were caused by metrizoate (2.78). These reactions caused discomfort and occasionally movements, with blurring of the films. Both contrast media provided sufficient information to enable an accurate radiologic diagnosis. PMID- 2980547 TI - A prospective comparison of iotrolan, iohexol and iopamidol for lumbar myelography. AB - The provisional results are presented of a comparative blind trial of iotrolan, iohexol and iopamidol for lumbar myelography. The aim of the trial was to assess the relative safety, tolerance and radiologic efficacy of the media. From the data available to date the incidence of side effects is similar for all three substances. Iotrolan does not provide specific imaging advantages. PMID- 2980548 TI - Cervical myelography with iohexol and metrizamide. A randomized double blind clinical study. AB - A randomized double-blind study with iohexol and metrizamide in cervical myelography was performed in 100 patients. The concentration of the contrast medium was 240 mg I/ml. The image quality was equal with both contrast media. Forty-two per cent of the patients receiving iohexol had side effects (headache 22%, nausea 6%, vomiting 2% and mental reactions 6%), in contrast to 80 per cent of the patients receiving metrizamide (headache 56%, nausea 34%, vomiting 24% and mental reactions 26%). The majority of side effects appeared within 6 hours after injection of contrast medium. Two patients had a late onset of symptoms - one in each group. Once appearing, the side effects tended to be equally severe. PMID- 2980549 TI - Magnetic resonance imaging of hemorrhagic conditions. AB - There is a stereotyped progression in the appearance of hemorrhage or thrombosis in magnetic resonance imaging performed at 1.5 tesla (T). This progression begins with low intensity on T2 weighted images (WI), and progresses to high intensity on T1 and T2 WI associated eventually, in some cases, with peripheral low intensity on T2 WI. Depending upon the etiology and location of hemorrhage or thrombosis the latter hypointensity on T2 WI may or may not be present. Hemorrhages occurring in regions of the brain not having an intact blood-brain barrier do not usually display persistence of low intensity. Regardless of etiology, consistent patterns in the evolution of hemorrhage may be observed on MR. PMID- 2980551 TI - The myeloneurogram. AB - Past attempts to visualize the internal structure of the spinal cord in vivo have been impeded by many factors including: the small size of the cord, the dense bony covering of the spine, and the likeness of tissue densities from one area to another within the cord. Delayed computed tomography is presently being used as a complement to iopamidol myelography to image the deep gray matter of the cord. This visualization is achieved by a differential gray-white matter enhancement, possibly due to a shielding effect of the white matter surrounding the gray matter, a difference in absorption rate between gray and white matter, or an increased rate of re-absorption of water-soluble contrast by the more highly vascularized deep gray matter. PMID- 2980550 TI - Iopamidol for intrathecal use in pediatric neuroradiology. AB - A clinical trial using iopamidol, a non-ionic contrast medium, in various iodine concentrations (300, 200 and 150 mg I/ml), was carried out in 947 children, aged between 1 day and 14 years, who underwent various neuroradiologic procedures: computed tomography (CT) cisternography (n = 307), CT ventriculography (n = 203), CT myelography (n = 153), and conventional myelography (n = 284). No significant variation in vital signs and neurologic status was recorded during and up to 72 hours after the examination. Psychic agitation was the major effect registered, being clearly related to the type of procedure used. A significant reduction of untoward side effects (about 50%) was observed using the lower concentrations of the medium. Iopamidol 150 mg and 200 mg represented the optimal concentration for CT enhancement, these concentrations reducing artifacts; the 300 mg I/ml concentration gave best results in conventional radiology. It seems that the lower concentrations of iopamidol (200 and 150 mg I/ml) can be usefully employed in pediatric neuroradiology, with preserved contrast and dramatically reduced untoward reactions. PMID- 2980552 TI - Low dose cervical CT myelography. How acceptable are adverse effects at this juncture? AB - When cervical myelography is required, the highest incidence of adverse effects usually supervenes. These effects are particularly important in patients with metastatic disease, post-cervical trauma and out-patients. Low dose hydrosoluble CT myelography imaging (300-500 mg I, total dose) can be accomplished by injecting the contrast medium when the patient is in the CT scanner via C1-2 puncture with a small needle (e.g. 25 gauge). Our method of accomplishing this was to use C-arm fluoroscopy performed with the patient either supine or prone and to transfer the patient with the needle in situ to the scanner. This was done with the patient on a portable exchangeable CT table top. Remarkably few adverse effects (transient mild headache in 2 of 22 patients) would appear to render this technique safe and useful. PMID- 2980553 TI - Iopamidol 150. A low iodine content non-ionic water-soluble contrast medium for CT myelocisternography. AB - Results of a clinical trial in CT myelocisternography performed with a new reduced concentration of iopamidol (150 mg I/ml) are reported. This compound is characterized by very low osmolality and osmotic pressure close to those of blood and CSF. Thirty patients were examined. Before, during and after the procedure pulse rate and blood pressure were recorded, EEG was taken and neurologic examination performed. After the examination no alteration in these parameters was recorded. The demonstration of the subarachnoid space and related structures was invariably adequate for diagnosis. In no case was a repeat study using a higher concentration of contrast medium needed. Mild reactions occurred only in three patients (10%). The authors conclude that iopamidol 150, owing to its safety and good imaging capacity, is useful in CT imaging of the subarachnoid space. PMID- 2980554 TI - Iotrolan. The first dimeric non-ionic contrast medium for the subarachnoid space. AB - Iotrolan was administered to 1400 in-patients and out-patients by intrathecal injection within the framework of the clinical investigation. It was used for studies of the lumbar, thoracic, cervical and intracranial areas. It was compared with metrizamide, iopamidol and iohexol in concentrations of 190, 240 and 300 mg I/ml in controlled double-blind trials. The minimum observation periods were four days including the investigation of clinicochemical, cardiovascular and electroencephalographic parameters, and CT. The incidence and severity of all side effects after Iotrolan were much lower than those after the non-ionic monomers. No epileptogenic incidents or symptoms of a psychosyndrome were observed in any of the cases--not even after prolonged retention in the intracranial fluid spaces. PMID- 2980555 TI - The effect of subarachnoid metrizamide and iohexol on cerebral glucose metabolism in vivo. AB - We studied the effect of metrizamide and iohexol on local cerebral glucose utilization (LCGU) using autoradiography. Metrizamide or iohexol was introduced into the subarachnoid space of rabbits and positioned over the right hemisphere for 3 or 6 hours. Using the unexposed (left) hemisphere as an internal control, we calculated relative changes in LCGU. In the three metrizamide-exposed animals the LCGU was decreased over the contrast media-exposed right temporoparietal cortex (L-R difference was positive). There was no significant changes in LCGU and no positive L-R difference after treatment with iohexol (three animals). We conclude that metrizamide does affect glucose metabolism in vivo, and this effect is dependent on localization of the contrast medium. The clinically safer drug, iohexol, in similar concentration and higher osmolality, did not decrease glucose metabolism. PMID- 2980556 TI - Paramagnetic enhancement of cerebral lesions with gadolinium-DTPA. AB - Magnetic resonance imaging (MRI) was performed on 30 patients with cerebral symptoms before and after intravenous gadolinium-DTPA (Gd-DTPA) administration. T1 and T2 weighted images were obtained in all patients. Fifty abnormalities were found in 27 patients. The post-contrast images revealed lesions not seen on pre contrast scans in 6 patients. Also, the post-contrast images showed a change in morphology of the lesions in 15 that provided additional information for assessing the abnormality. The Gd-DTPA did not obscure any of the lesions seen on pre-contrast scans. Gd-DTPA improves conspicuity, helps characterize and delineate the extent of lesions and increases the sensitivity for detection of cerebral abnormalities. PMID- 2980557 TI - Importance of oxygenation in the appearance of acute subarachnoid hemorrhage on high field magnetic resonance imaging. AB - The ability to detect acute hemorrhage by magnetic resonance (MR) is related to the oxygen saturation of hemoglobin. This experiment measured the relaxation times of solutions of cerebrospinal fluid containing a 5 per cent hematocrit at various pO2's on a 1.4 tesla (T) MR imaging system. The results demonstrate that the state of oxygen saturation of hemoglobin determines the extent of T2 relaxation at this field strength. The T2 relaxation rate varies quadratically with the concentration of deoxyhemoglobin. There were no significant changes in the T1 relaxation rate with variations in pO2. These findings may, in part, explain the inability of MR to detect subarachnoid hemorrhage, and the MR appearance of blood in intratumoral hemorrhage, hemorrhagic cortical infarction and neonatal hemorrhage. PMID- 2980558 TI - Magnetic resonance imaging of intracranial tumors using gadolinium-DTPA. Initial experience with fast imaging. AB - One hundred and fifty gadolinium(Gd)-DTPA studies in patients with intracranial tumors were performed. In meningiomas (n = 37) Gd-DTPA is useful to delineate isointense tumors. In acoustic neuromas (n = 16) Gd-DTPA helps to identify intracanalicular tumor tissue. In pituitary adenomas (n = 16) para- or suprasellar tumor extension can be better appreciated after Gd-DTPA injection. In glioblastomas (n = 23) Gd-DTPA provides data on differential diagnosis through the characteristic shape of enhancement. In a variety of tumor types the high contrast between enhancing tumor tissue and adjacent structures facilitates therapy planning. Fast imaging sequences now allow for dynamic Gd-DTPA studies. The time-course of enhancement may increase specificity of magnetic resonance imaging. PMID- 2980559 TI - Demonstration of meningeal contrast enhancement on magnetic resonance imaging. AB - Fourteen cases are described in which surface enhancement of the brain or spinal cord was observed on magnetic resonance (MR) scans performed using the paramagnetic intravenous contrast agent gadolinium-DTPA. The sign was seen in cases of meningioma, extradural haematoma, neurosarcoidosis and after surgery and radiation therapy for malignant tumour. This sign is demonstrated on MR with increased sensitivity compared with computed tomography, and is useful in indicating dural attachment of meningiomas. It may also be of value in the presence of meningeal inflammatory changes from a variety of causes. PMID- 2980560 TI - Gadolinium-DTPA in the magnetic resonance evaluation of the postoperative patient. Work in progress. AB - The application of gadolinium-DTPA to magnetic resonance scanning has been demonstrated to have considerable potential. Because the evaluation of the post operative patient is particularly difficult for magnetic resonance due to the non specific appearance of surgical trauma and edema, we attempted to formulate some preliminary impressions regarding the use of gadolinium-DTPA in this setting based on our experience with 15 patients studied at varying intervals following surgery, from 2 days to 8 years. Specifically, the time course and appearance of normal post-surgical enhancement was found to exhibit certain patterns. Recognition of these patterns may allow differentiation from pathology and may result in gadolinium-DTPA adding useful information in evaluating these patients. PMID- 2980562 TI - Treatment of direct carotid cavernous sinus fistulae. Various therapeutic approaches and results in 148 cases. AB - From 1974 to 1986, 148 patients with carotid cavernous fistula (CCF) were evaluated for intravascular therapy. Four patients died from hemorrhage before treatment could be instituted and the CCF closed spontaneously in 5. Therapeutic approaches which resulted in complete occlusion in the remaining 139 cases were transarterial in 118, transvenous in 15 and external compression of the carotid artery and jugular vein in 6. The current treatment of choice of the direct CCF is intravascular embolization using detachable balloons, particulate emboli or liquid adhesive agents to occlude the CCF while attempting to preserve the carotid artery. In 15 patients it was technically too difficult to use the transarterial approach. The patients were therefore treated from a transvenous approach including access via the femoral vein, superior ophthalmic vein, intraoperatively from the inferior petrosal sinus or direct puncture of the cavernous sinus. Embolic agents used included detachable silicone balloons, steel minicoils, particulate emboli and isobutyl-2-cyanoacrylate. In 14 of these 15 patients total obliteration was achieved with marked improvement in symptoms. Complications occurred in 3 patients including perforation of the cavernous sinus resulting in subarachnoid hemorrhage, delayed pontine hemorrhage from subtotal occlusion of the fistula and transient increased proptosis. PMID- 2980561 TI - Action of gadolinium complexes on different enzyme systems. AB - Gadolinium complexes have two advantages: their toxicity is very low and the paramagnetic properties are adequate enough to give a useful signal with magnetic resonance imaging (MRI). The effects of several MRI contrast media on the activity of two brain enzymes, glutamate decarboxylase (GAD) and cholineacetyl transferase (ChAT), were investigated. The study was performed on homogenates of rat striatum with variable concentrations of gadolinium sulphate, EDTA, DTPA, DOTA, Gd-EDTA, Gd-DTPA and Gd-DOTA (from 10(-2) mol/l to 10(-9) mol/l. Gadolinium in its ionic form and the chelators of cation agents such as EDTA, DTPA and DOTA inhibit GAD, a calcium dependent enzyme, whereas they have no inhibiting effect on the activity of ChAT, a calcium independent enzyme. Gadolinium complexes inhibit the GAD activity by about 15 per cent whereas they do not modify that of ChAT. The complex form of gadolinium has the advantage of retaining its paramagnetism while drastically lowering its toxicity. PMID- 2980563 TI - Closure of carotid cavernous sinus fistulae by external compression of the carotid artery and jugular vein. AB - From 1974-1986, 152 patients with carotid cavernous sinus fistulae (CCF) have been evaluated. Progressive closure of both dural and direct types of CCF have been noted utilizing intermittent external manual compression of the cervical carotid artery and jugular vein. In a group of 71 patients in whom this treatment was attempted, we have observed that 7 of 23 patients (30%) with dural CCF, and 8 of 48 patients (17%) with direct CCF had complete closure of their fistulae with no recurrence either clinically or at angiography done one year later. Closure occurred at varying times, from several minutes to 6 months (mean 41 days) following compression therapy. In those patients with CCF without rapidly progressive visual deterioration, cerebral ischemia, or other complicating factors, we recommend this technique with serial clinical follow-up and angiography before more definitive therapy is employed. PMID- 2980564 TI - Diagnosis and endovascular therapy of vascular lesions in the cavernous sinus. AB - One hundred and twenty-eight lesions of the cavernous sinus were diagnosed and treated by endovascular embolization alone or combined with postembolization surgery. All patients presented with acute or subacute cavernous sinus syndrome. A complete angiographic evaluation included bilateral internal and external carotid and vertebral angiography. A total of 88 carotid-cavernous (c-c) fistulae (68 traumatic and 20 spontaneous) and 40 giant aneurysms were demonstrated. A complete anatomic cure or satisfactory clinical improvement was seen in 67 of 68 traumatic fistulae, in 14 of 20 spontaneous c-c fistulae and in 38 of 40 giant intracavernous aneurysms. An overall 15 per cent immediate and 2.7 per cent long term morbidity with one death was observed. The technical and clinical complications involving endovascular therapy of each specific intracavernous vascular lesion are analyzed. PMID- 2980565 TI - Intravascular laser coagulation of experimental aneurysms. AB - Rebleeding after aneurysmal subarachnoid hemorrhage has a 45 per cent mortality rate. The peak incidence is within 24 hours of the initial bleed. During the ensuing 2 weeks 20 per cent of patients with suffer a second hemorrhage. To avoid this, surgical treatment must be instituted promptly but, logistically, this is often difficult to implement. To this end, we have devised an intravascular laser catheter technique that may provide expeditious treatment for intracranial aneurysms. We have coagulated experimental aneurysms in the rabbit carotid artery using laser energy transmitted through an intra-arterial optical fiber. The special fiber tip was positioned in the aneurysm by a catheter introduced from the femoral artery. Laser power of 750 mW and an exposure of 15 seconds caused aneurysm ablation leaving the feeding artery patent. To date, 4 experimental aneurysms have been occluded by this technique and long term histologic results are pending in this ongoing study. PMID- 2980566 TI - Cervical carotid artery aneurysms and pseudoaneurysms. Treatment by balloon embolization therapy. AB - The treatment of high cervical carotid artery aneurysms and pseudoaneurysms is often difficult due to relative surgical inaccessibility. We are currently managing such lesions by detachable balloon embolization therapy. In 7 patients with acute cervical carotid pseudoaneurysms, the carotid artery was occluded by a trapping procedure above and below the lesion using detachable silicone balloons. In 3 patients with aneurysms that had well organized walls, a detachable balloon was placed directly within the aneurysm, inflated to exclude it from the circulation, and then detached. The carotid artery was therefore preserved. PMID- 2980567 TI - Intravascular detachable balloon embolization of intracranial aneurysms. Indications and techniques. AB - The treatment of intracranial aneurysms from a transvascular approach, with preservation of the parent vessel is now being performed in selected cases. From a transfemoral approach, a silicone detachable balloon is flow-directed up the carotid or vertebral-basilar artery, guided directly into the aneurysm and detached. The aneurysm is thus excluded from the circulation and the parent artery is preserved. Thus far, this technique has been successful in treating aneurysms in the cavernous carotid, carotid ophthalmic, posterior communicating, distal basilar, and posterior cerebral artery distributions. The indications and techniques of detachable balloon embolization therapy are presented. PMID- 2980568 TI - Giant carotid aneurysms. Treatment by detachable balloons. AB - Six patients with giant aneurysms of the internal carotid artery have undergone detachable balloon occlusion of the aneurysm. The aneurysm lumen was successfully obliterated in all cases by either occluding the neck of the aneurysm, trapping the aneurysm or occluding the parent vessel just proximal to the origin of the aneurysm. One patient, who was in poor clinical condition pre-operatively suffered a hemiplegia 6 hours after the procedure. No other permanent complications occurred. This procedure offers a satisfactory alternative to surgery in selected patients. PMID- 2980569 TI - Superselective catheterization using very flexible, formed catheters. AB - Superselective catheterization of even very tortuous vessels is possible, with very low risk of arterial spasm and plication of the catheter, when very flexible catheters are used. The thermoplasticity as a new additional quality of very flexible catheters offers new possibilities to guide such catheters. Thus, the catheter extremity may be curved in order to overcome difficult anatomic pathways, or a bulb-shaped dilatation of the catheter tip can be achieved in order to increase the flow-dependent guidance of the catheter. PMID- 2980570 TI - Radiosurgery using a 4MV linear accelerator. Technique and radiobiologic implications. AB - In the authors' technique, the stereotactically localized target is fixed to the isocenter of a 4 MV linear accelerator. The irradiation is carried out along 9-17 non-coplanar arcs distributed on a 160 degrees cylindrical sector. High doses (10 50 Gy) are delivered in one or two sessions. From 1982, 65 patients have been treated (shortest follow-up 6 months). The dose was chosen according to the pathology and to the volume of the lesion. Good clinical results have been obtained in low-grade gliomas, acoustic neuromas, arteriovenous malformations, and other selected types of intracranial lesions. Therapeutic effects in terms of clinical condition and size of the lesions have been plotted in relation to time elapsed and dose employed. PMID- 2980571 TI - Stereotactic radiation therapy in arteriovenous malformations and brain tumors using the Fixster system. AB - In brain tumours, stereotactic radiation therapy, performed in a single high dose (SHD) or fractionated (FSR), gives better results than conventional methods. In arteriovenous malformations (AVM), radiosurgery is a well-defined alternative to neurosurgery and embolization, if the malformation is not wider than 3 cm. In large AVMs, a successful combined technique of radiosurgery and FSR has been developed. In a series of 110 patients the experience proved that the atraumatic Greitz-Bergstrom stereotactic system is appropriate. PMID- 2980572 TI - Stereotactic radiation therapy of intracranial arteriovenous malformations. AB - Twenty-six patients with large intracranial arteriovenous malformations (AVM), not suited for open surgery or radiosurgery, have been treated by fractionated stereotactic radiation therapy using a linear accelerator. The preliminary results are based on follow-up angiography one and a half years after treatment, and in 5 cases also 5 years after treatment. In 2 cases a complete obliteration of the AVMs has occurred, and in the majority of the remaining cases a reduction of the size of the lesion has been obtained. Further development of the method is needed in order to improve the results. Today treatment with this technique is only recommended in cases which cannot be treated with other methods. PMID- 2980573 TI - Magnetic resonance imaging of stereotactic radiosurgical lesions in the internal capsule. AB - Confirmation of radiosurgical lesions in white matter has earlier been difficult to obtain, which has hampered the evaluation of clinical outcome in relation to the site and the size of the lesions. In this investigation 7 consecutive patients subjected to bilateral stereotactic gamma capsulotomy for intractable anxiety disorders were re-examined several years after treatment. The protocol included both magnetic resonance imaging (MRI) and computed tomography (CT) interpreted under blind conditions and prospective psychiatric evaluations performed by two independent evaluators who had not been involved in the selection and the treatment of the patients. The lesions were clearly visible with MRI in all patients who improved after treatment. MRI proved to be more accurate than CT both in detecting and in defining the size and the configuration of the lesions. The extent of the tissue reaction following the irradiation seemed to be best defined in T2 weighted images. A high correlation (p less than 0.01) was obtained between ratings of clinical outcome and radiosurgical precision as reflected by MRI. It is concluded that MRI may provide the clinician with more information than CT does. In the future MRI may also facilitate the determination of a radiation threshold value for white and grey matter lesions and provide a more detailed knowledge of the time course of the development of such lesions. PMID- 2980574 TI - Improved technique of bucrylate embolisation in brain arteriovenous malformation. The use of additional temporary balloon occlusion. AB - Problems currently limiting the success and safety of calibrated leak balloon embolisation include suboptimal occlusion of malformation bed due to multiple or inaccessible pedicles and neural deficits due to paradoxical embolisation of normal brain. Analysis of preliminary calibrated leak balloon angiograms supported by temporary balloon occlusion of other pedicles yields insights into the dynamics of blood flow to anterior malformations. Strategies have been devised using temporary balloon occlusion to improve filling of malformation bed and reduce paradoxical embolisation of eloquent brain. PMID- 2980575 TI - Endovascular treatment with isobutyl cyano acrylate in patients with arteriovenous malformation of the brain. Indications, results and complications. AB - Between December 1984 and May 1986 67 patients with arteriovenous malformation (AVM) of the brain have been treated with endovascular occlusion using isobutyl cyano-acrylate at the Lariboisiere Hospital. Six patients were completely cured, i.e. with complete occlusion of the malformation. In 21, reduction of the size of the AVM was obtained, permitting surgery or radiation therapy not considered possible before endovascular intervention. In 33 cases a partial cure was achieved, but additional embolization is needed. In 3 cases the treatment was interrupted. Four patients died. The overall frequency of complications was in this material around 10 per cent. PMID- 2980576 TI - Ethyl 2-cyanoacrylate as an embolic agent for cranial arteriovenous malformations. An experimental study. AB - Physical properties of ethyl 2-cyanoacrylate (Aron Alpha) were studied in comparison with isobutyl 2-cyanoacrylate (IBCA). Aron Alpha is characterized by high viscosity, a short polymerization time, low spreadability and low fragmentability. On the other hand, IBCA had low viscosity, a relatively long polymerization time, high spreadability and high fragmentability. Based on these findings, it is suggested that IBCA is suitable for transcatheter embolization, and Aron Alpha for intraoperative embolization. Mixture of IBCA with 5% Aron Alpha suppressed the spreadability and the fragmentability of IBCA, resulting in reduction of distal migration of IBCA into the venous system. PMID- 2980577 TI - Valved latex balloons and water-soluble contrast opacified polymer. Simple detachable balloon system. AB - Latex balloons of different shapes, sizes and volume, incorporating a simple valve mechanism, were produced cheaply. These balloons are ethylene oxide sterilized and are available on 'the shelf' for use at any time. A contrast soluble polymer consisting of 2 hydroxy-ethyl-methacrylate, a cross linker, Iopamidol and water-soluble free radical curing system has been developed. The polymer sets in 10 minutes and there is no 'dead volume' problem. The balloons and polymer were evaluated in dog arteries, including histologic studies and have been subsequently used in 7 patients without any complications. PMID- 2980578 TI - Magnetic resonance characterization of non-flowing intravascular blood. AB - We eliminated the effects of flow on the magnetic resonance (MR) appearance of intraluminal blood by imaging the superficial veins in the arms and legs of 3 volunteers during venous occlusion with tourniquets. T1, intermediate and T2 weighted transaxial images were obtained using a multisection spin-echo technique at 1.5 T. The images were analyzed by visual inspection and by region of interest pixel intensity measurements. The intensity of non-flowing blood was compared with muscle, fat and a tube of water taped to the skin. We determined that non flowing, unclotted blood has intermediate T1 characteristics and long T2 characteristics. On T1 weighted images, non-flowing blood is of greater signal intensity than water or muscle, but less than fat. On T2 weighted images, non flowing blood is of greater signal intensity than fat and muscle, but slightly less than water. These results may help explain the signal observed with very slow intravascular flow and hyperacute intracranial hematomas, where intermediate T1 and long T2 relaxation characteristics are reported. PMID- 2980580 TI - Minisymposium on pediatric neuroradiology. PMID- 2980579 TI - Ventriculocystostomy. A new method of percutaneous intracranial shunting. AB - A new method of treatment for arachnoid intracranial cysts is described, involving introduction of a catheter which enables fluid passage from the cyst into the ventricular system, and its resorption. Successful drainage has been achieved in 17 patients. PMID- 2980581 TI - Genetic brain malformations recapitulate phylogeny. AB - It is generally difficult to establish whether a congenital brain malformation has a genetic basis or is due to other factors. A thesis has been developed, that brain malformations with a genetic basis recapitulate phylogeny. In order to validate this thesis a retrospective CT investigation of brain dysmorphology was carried out in 50 cases of dysgenesis of the corpus callosum, 20 cases of the holoprosencephaly complex, 25 cases of Dandy-Walker malformation, and 50 cases of miscellaneous brain anomalies. This group was compared with 100 neonates with brain damage from acquired causes. The results strongly suggest that a brain malformation that recapitulates phylogeny is very likely to be based on genetic aberration. PMID- 2980582 TI - Ultrasonography and computed tomography in midline cerebral malformations. AB - Ultrasonography (US) has become an important method for the investigation of neonatal cerebral disorders. Its role in the diagnosis of cerebral malformations has been well illustrated in the recent literature. This report presents 16 children with midline malformations (of 78 neonates and children with cerebral abnormality diagnosed with US) who were also examined with computed tomography (CT). Among the 16 children there were 2 with lipoma of the corpus callosum, 1 neonate with aneurysm of the vein of Galen, 2 with agenesis of the corpus callosum, 1 newborn with a large cyst in the quadrigeminal plate cistern, 3 with a Dandy-Walker malformation, 5 with a posterior fossa cyst, and 2 with a Chiari II malformation. The diagnostic accuracy of ultrasonography in midline malformations and the role of computed tomography for further investigation is emphasized. CT was necessary after US mostly in hyperechogenic lesions such as lipomas and partly thrombosed aneurysms of the vein of Galen, while in the anechogenic cystic lesions US alone was satisfactory. PMID- 2980583 TI - Lumbo-peritoneal shunt malfunction. A new, simple and reliable CT sign. AB - Sixty CT scans in 31 patients who underwent lumboperitoneal shunting for communicating hydrocephalus showed that the size of the ventricles did not represent a good indicator of shunt malfunction. Instead, we discovered that the size of the basal cisterns around the brain stem enabled us to predict blockage earlier and more reliably. In a well functioning shunt, the basal cisterns are usually not visualized. In children with clinical shunt malfunction the cisterns dilate and become visible again. This occurs earlier and more frequently than ventricular enlargement. We therefore conclude that visible cisterns in association with persistent symptoms of malfunction are more reliable predictors of a true blockage that requires shunt revision than serial studies of ventricular size. PMID- 2980584 TI - Magnetic resonance imaging of the pediatric spinal cord and canal. AB - Of 105 pediatric patients studied on a 1.5 tesla MR unit for clinical suspicion of spinal disease 69.5 per cent of studies were positive. Magnetic resonance imaging (MRI) was successfully performed on all patients, from newborn up. MRI replaced invasive studies such as myelography and intrathecally enhanced computed tomography (CT) in all disease categories except for subarachnoid drop metastasis, spinal arteriovenous malformation and arachnoiditis. Conventional CT and radiography of the spine were complementary in disease processes where bony detail was important. PMID- 2980585 TI - Gliomas in children following radiation therapy for lymphoblastic leukemia. AB - Four children treated for acute lymphoblastic leukemia developed intracranial gliomas between 40 and 120 months following multiagent chemotherapy, prophylactic whole brain irradiation, and intrathecal Methotrexate. The diagnosis of glioma was confirmed in each case with biopsy or autopsy. These children were part of a larger series of 73 patients with acute lymphoblastic leukemia treated at Indiana University receiving 24 Gy as per the protocol guidelines of the Children's Cancer Study Group. Currently 42 patients of the original 73 children survive in continuous remission. PMID- 2980586 TI - Radiology of orbital masses in children. AB - Ninety-three children (103 orbital tumours) have been examined. Typical appearances were noted in one half of the cases. Most of the optic nerve gliomas, rhabdomyosarcomas, metastatic neuroblastomas and all the dermoids were typical. The most variable appearance was seen in vascular malformations which sometimes could be confused with typical lesions of other histologic type. PMID- 2980587 TI - Computed tomography and magnetic resonance imaging in children with rare movement disorders. AB - Both CT and MR can aid in the differentiation between benign and less favorable pediatric movement disorders. However, MR is the more sensitive imaging modality and thus preferable. This was demonstrated in 18 patients with conditions ranging from idiopathic dystonias to aminoacidopathies, Leigh's disease, and Hallervorden Spatz disease. In the latter three entities characteristic lesions were shown in the basal ganglia, although not consistently. A specific diagnosis seems possible in Leigh's disease, provided the clinical setting is appropriate. PMID- 2980588 TI - Radiologic diagnosis of periventricular leukomalacia. AB - Fifteen infants with clinical evidence of periventricular leukomalacia, i.e. spastic diplegia or quadriplegia and premature birth were studied. CT scans of the brain demonstrated: 1) reduction in quantity of periventricular white matter, 2) deep and prominent sulci which abutted the ventricles without interposed white matter and 3) ventriculomegaly with irregular outline of the lateral ventricles. The location and severity of abnormalities on CT scans correlated well with neurologic handicap and the known anatomic location of periventricular leukomalacia, thus demonstrating the diagnostic value of CT scanning for periventricular leukomalacia during later infancy. PMID- 2980589 TI - Perinatal hypoxic-ischaemic syndrome. Diagnostic and prognostic value of computed tomography. AB - Sixty-two full-term neonates with hypoxic-ischaemic encephalopathy were studied. Computed tomography (CT) was performed during the first week of life and the clinical and radiologic findings were correlated. In 57 cases (92%) CT was abnormal. Subarachnoid haemorrhage (SAH) was present in 53 cases, oedema in 23 cases and other lesions in 7 cases. Twenty-two cases had associated SAH and oedema. It was concluded that SAH and/or oedema has a good prognosis. When neurologic involvement is severe and CT shows signs of necrosis, the prognosis is less favourable. PMID- 2980590 TI - Development of the new cerebral angio-CT technique and magnetic resonance angio imaging. AB - Using dynamic scanning, multiple cerebral computed angiotomography scans were obtained after a single bolus injection of contrast medium. The following three different techniques and their clinical applications were investigated. 1) Overlapping scanning was applied to detect unruptured intracranial aneurysms. 2) Sequential scanning was mainly applied for larger lesions such as isodense chronic subdural hematomas. 3) Two-step contrast-enhanced scanning was useful for investigating the relationship between tumor and surrounding vessels. Cerebral magnetic resonance angio-imaging was recently investigated as a noninvasive procedure using spin-echo magnetic resonance imaging. PMID- 2980591 TI - Topographic cerebral blood flow mapping in children with ischemic cerebrovascular diseases using xenon-enhanced computed tomography. AB - A non-invasive technique using xenon-enhanced computed tomography (Xe-CT) for obtaining topographic regional cerebral blood flow (rCBF) maps has been developed. During the past 3 years, the Xe-CT has been performed in 22 children with ischemic cerebrovascular diseases, including 17 cases with moyamoya disease. In moyamoya disease, rCBF maps revealed low flow in the bilateral frontal and temporal cortices, and high flow in the thalamus and putamen on both sides. Other diseases showed low perfused areas. Xe-CT can be easily performed even in infants and children and it gives valuable information that may be of clinical use. PMID- 2980592 TI - Comparison of cerebral blood flow measurements by xenon computed tomography and dynamic brain scintigraphy in clinically brain dead children. AB - Because of the controversy over applying adult clinical and laboratory criteria to children suspected of brain death, xenon computed tomographic cerebral blood flow (XeCTCBF) and near simultaneous scintigraphy were studied in 10 children. Six met the clinical criteria, one of whom had three indeterminate scintigrams with only central flow on XeCTCBF. Another had considerable flow with scintigraphy and XeCTCBF. One with clinical and autopsy brainstem death but preserved cortical function also had flow on both. This preliminary study establishes that brain scintigraphy is occasionally hypersensitive to insufficient cerebral flow and promises more accurate early assessment with XeCTCBF. The expectation is that XeCTCBF can refine the criteria for the earlier establishment of brain death. PMID- 2980593 TI - Changes in frontal lobe size with age. A CT study in children. AB - A series of linear measurements on computed tomography of the brain was performed in 481 children (from newborns up to 5 years of age). Each measurement increased with age in infancy and early childhood. In addition to the commonly used linear measurements, a new index has been devised. This index equals the ratio between the sagittal diameter of the brain and the sagittal midline distance from the anterior border of the brain to a line tangential to the anterior horns. All measurements were made on the scan at the level of the foramen of Monro. This index was called the frontal force index (FFI), because it seemed to reflect the morphologic change in the frontal lobes. The functions related to the frontal lobes, i.e. cognitive functions, develop rapidly during infancy and early childhood. In normally or subnormally developed children the FFI increased during early infancy and decreased in severely retarded children. The FFI is obviously useful in the evaluation of the development of the frontal lobes. PMID- 2980594 TI - Computed tomography and dynamic cerebrospinal fluid studies in the evaluation of extracerebral fluid collections in infants with hydrocephalus. AB - Infants presenting with an increasing head circumference were subjected to computed tomography (CT). A sub-group of 15 were identified in whom the CT scan disclosed varying degrees of ventricular enlargement together with bilateral extracerebral fluid collections. A method is described and illustrated for determining in which compartment the fluid lies. In most of the infants, non ionic water-soluble contrast medium was injected intrathecally via lumbar puncture and directed intracranially by gravity. Subsequent CT scans monitored the passage of contrast through the subarachnoid spaces of the brain from which it was possible to determine whether there was any communication between them and the extracerebral fluid. In the remaining children, contrast medium was injected directly into the extracerebral fluid and CT was again used to demonstrate any communication between this fluid and the basal cisterns. PMID- 2980595 TI - The bifrontal fluid collection area and other cerebrospinal fluid spaces at computed tomography in children. Its variations with increasing age and pathologic conditions. AB - A total of 592 computed tomography examinations of the brain in children were studied from the aspect of age and development. The series was divided into 3 groups with respect to neurologic development: 1) normal without neurologic symptoms, 2) normal or subnormal with neurologic symptoms, and 3) moderately to severely retarded. For evaluation, a method of visual grading was applied for each cerebrospinal fluid space. Concerning the ventricular system, the most significant changes in size with increasing age were noted in the lateral ventricles. There were no significant changes in the third and fourth ventricles. Among the three groups, the enlargement was most prominent in group 3 and less prominent in groups 1 and 2. Concerning the extraventricular spaces, the enlarged bifrontal fluid collection area, the sylvian fissure, the interhemispheric fissure, and the cerebral sulci vanished with increasing age. Similarly to the ventricular system, the enlargement was most prominent in group 3, and less prominent in groups 1 and 2, above the age of 1 year. The relation of these results to age and developmental factors are discussed. PMID- 2980596 TI - Benign hydrocephalus in infants. A computed tomographic and clinical correlative study. AB - During the past 3 years we studied benign hydrocephalus in 7 infants, who presented symptoms of macrocrania and mild psychomotor retardation. Computed tomography (CT) revealed dilatation of CSF space resembling brain atrophy or subdural effusion. These children were observed clinically and were found gradually to restore to normal with regard to CT changes, as well as clinically. These patients were compared with another group of patients with subdural effusion or hematoma who needed tapping or shunting. PMID- 2980597 TI - Hyperfrontal distribution of regional cerebral blood flow and vascular CO2 reactivity in normal subjects and disturbances in ischemic cerebrovascular disorders. AB - The distributions of regional cerebral blood flow (rCBF) and CO2 reactivity were estimated in normal subjects and in patients with ischemic cerebrovascular disorders, using the 133Xe inhalation method. In the normals, hyperfrontal and hyper-fronto-parietal distributions of rCBF and CO2 reactivity were observed. In the patients, the differences in mean rCBF values between anterior and posterior regions decreased in the affected hemisphere. Comparison between the hemispheres revealed that the anterior regions had a decreased mean rCBF value in the affected hemisphere. The CO2 reactivity in the patients was generally decreased in both hemispheres, but not significantly. The originally existing, different distributions of rCBF and CO2 reactivity in normals must be considered in the evaluation of disturbances of vascular reactivity in patients with cerebral disorders. PMID- 2980598 TI - Pediatric surgical neuroangiography. A multicentre approach. AB - Adult patients who require sophisticated embolization procedures are usually referred to centres where such expertise is available. Because of the special circumstances in the pediatric age group, this type of referral is often not feasible. The authors describe a model in which multiple pediatric institutions have access to sophisticated interventional neuroradiologic procedures through the collaboration of a group of neuroradiologists. The preliminary results are encouraging and appear to indicate that embolization in the pediatric age group can be safe and reliable if performed by well trained teams. PMID- 2980599 TI - Computed tomography of the lumbar spine. Experience with an Expert System in computer aided diagnosis. AB - The authors report their experience in the formulation of an Expert System for computer aided diagnosis in the examination of the lumbar spine with computed tomography. A large number of steps are identified in the process of diagnostic evaluation of CT examinations; about 80 decisional rules are considered and arranged in five 'areas'. The Expert System under study is a first proposal, intended for use by radiologists with some basic training. PMID- 2980600 TI - The critical role of 3-D CT reconstructions for defining spinal disease. AB - Three dimensional processing of routine CT images has previously been applied to osseous related maxillo-facial and spinal disorders. Two groups of patients, 25 with substantial spinal trauma and 25 with 'failed back' syndrome had 2-D and 3-D like displays processed by the Cemax 1000 system. The goal was to objectify whether the adjunct of 3-D imaging was truly valuable diagnostically. All images were recorded on 35 mm slides and projected both randomly and as an organized case; intra- and interpersonal evaluations were made. 3-D imaging in 19 of the 25 (76%) trauma patients disclosed additional diagnostic information which was considerably important to both the neuroradiologist and the referring surgeon. In the 'failed back' group, the 3-D images showed supplementary information in 15 of 25 (60%) cases. 3-D displays were usually in color showing complete regional information obtained from high resolution, medium thickness (4 mm) CT slices with minor overlapping (1 mm). The displays were optimized to the plane best defining the pertinent osseous and joint morphology; this included variably rotated and sometimes hemisected views. The images presented here are static, however, when viewed rapidly or by dynamic rotation, the regional morphology results in a highly graphic 3-D presentation. PMID- 2980601 TI - Three-dimensional interactive analysis of craniofacial and spinal computed tomography. AB - Three-dimensional interactive display of CT data from 23 cases of craniofacial and spinal pathology using a solids processing computer system was compared with conventional two-dimensional CT display. Three-dimensional display gave a more complete perspective of complex displacement patterns of facial and spinal fractures, and more clearly defined surface anatomy of osseous tumors and malformations. In 7 cases however, processing algorithms for three-dimensional display caused subtle anatomic features and non-displaced fractures to be obscured. These false negatives were clinically significant in cases of petrous bone pathology and in non-displaced spinal fractures that affected stability. PMID- 2980602 TI - Magnetic resonance imaging of the lumbar spine. A comparison with computed tomography and myelography. AB - Two hundred and fifty patients were referred for magnetic resonance imaging (MRI) of the lumbar spine. Computed tomography (CT) and MRI were performed in 50 patients and MRI, CT and myelography in 20 patients. Twenty patients had surgical confirmation of the imaging studies. MRI was best for demonstrating degenerated discs. MRI was better than CT for demonstrating disc bulge without herniation, and MRI was slightly better for herniated disc demonstration than CT. Myelography did not demonstrate degenerated discs. PMID- 2980603 TI - Assessment of the relative impact of magnetic resonance imaging on the evaluation of rheumatoid disease of the cervical spine. AB - A study was made of 15 patients with rheumatoid involvement of the neck in whom neurologic symptoms were of sufficient severity that operative intervention was being planned. Magnetic resonance imaging (MRI) was performed in all patients and compared with conventional radiography, conventional tomography, myelography and computed tomography (CT) assisted myelography. MRI clearly demonstrated any deformity or compression of the cord and was able to show the effects of flexion and extension of the neck. It is concluded that MRI could replace myelography for defining cord compression. If details of atlanto-axial alignment and bony outline of the spinal canal are required then CT will provide the appropriate images. PMID- 2980604 TI - High field strength magnetic resonance of extra-axial spinal pathology. AB - High field strength (1.5 T) surface coil magnetic resonance imaging (MRI) of the spine was performed on 560 patients over a 9 month period of time. A total of 220 patients with extra-axial pathology including spinal stenosis, spondylosis, herniated disc, traumatic, inflammatory, vascular, developmental and tumorous lesions were included in this initial spine MRI experience. Results of these studies clearly indicate an expanded role of increasing importance for MRI performed at high field strength in the evaluation of spinal extra-axial pathology compared with the information that has been obtained from examinations performed at lower field strengths. Studies obtained at 1.5 T may obviate the need for myelography in many cases. PMID- 2980605 TI - CT findings in the vertebral column in the radicular lumbosacral syndrome. AB - An analysis of the CT findings in 150 patients with the radicular lumbosacral syndrome showed that 98 of them (65.3%) were suffering from intervertebral disc prolapse or protrusion. In 44 of these patients the finding of a prolapsed disc was combined with major spondylarthrosis while 20 others had marked spondylarthrosis or spondylosis with no apparent disc prolapse. Other findings included metastatic destruction of the vertebra (4 cases), fractured vertebra (2 cases), osteoid osteoma (1 case), and discitis (1 case). Slight or major asymmetry was found in 94 per cent, and a high incidence (38.8%) of the atypical rotatable type of intervertebral articulation was noted. The high percentage seems to confirm the hypothesis, according to which the rotatable type of intervertebral union is one of the predisposing factors for the development of the radicular lumbosacral syndrome. PMID- 2980606 TI - Magnetic resonance imaging of the spine in adolescents. AB - The appearances of the spine in 14 adolescents with symptomatic intervertebral disc herniations and in 20 asymptomatic controls were compared at magnetic resonance imaging. Abnormality in the signal from the nucleus pulposus of one or more discs was present in all the patients, while only 4 of the 20 controls had any abnormal discs. Ten of the 14 patients had multiple-level disc abnormality. Five had three abnormal discs and five two-level abnormality. This suggests that there was an underlying diathesis in these patients towards the development of disc herniation. PMID- 2980607 TI - Radiculopathy in spondylosis/spondylolisthesis. The laminar/ligamentum flavum process. AB - Based upon an anthropologic and clinical material a specific bone formation, the ligamentum flavum process (LLFP), is described. The process is present in up to 87 per cent at a lytic site and appears to be the cause of root compression in 68 per cent of the patients with such a skeletal abnormality. LLFP is appreciated in axial CT and should be considered in planning possible surgical intervention. PMID- 2980608 TI - Dynamic studies of cervical spinal canal and spinal cord by magnetic resonance imaging. AB - In 15 cases dynamic studies of cervical spinal cord and canal in flexion and extension were performed by magnetic resonance imaging (MRI). In addition measurements of the complete spinal cord were made in 5 cases. As compared with extension the cervical spinal canal and spinal cord lengthen 12 mm in average during flexion, whereas the spinal canal lengthens 28 mm in average, which means a difference of about 15 mm. We feel that these results indicate that adverse mechanical tension may occur in the cervical spinal cord during flexion. The impact of these results on surgical treatment for chronic cervical myelopathy is discussed. PMID- 2980609 TI - Extracranial/intracranial anastomosis in therapeutic balloon occlusion of the internal carotid artery. AB - Occlusion of the internal carotid artery with a detachable balloon was attempted or achieved in 19 patients suffering from a carotid aneurysm or dural arteriovenous malformation. An extracranial/intracranial anastomosis was performed prior to the occlusion in 11 patients, 4 of whom developed complications as did one of the 8 patients without an anastomosis. Although the morbidity of the procedure might have been greater without an arterial bypass a comparison of the two groups of patients lends no support for this view. PMID- 2980610 TI - Magnetic resonance imaging of spinal cord hemangioblastomas and arteriovenous malformations. AB - Eight patients with either spinal cord hemangioblastomas (n = 5), spinal arteriovenous malformations (AVM) (n = 2) and intramedullary hematoma (n = 1) were studied by magnetic responance (MR) imaging. All hemangioblastomas, one AVM and the intramedullary hematoma were identified by MR. Compared with other neuroradiologic procedures, MR provided important complementary information in the diagnostic management of spinal hemangioblastomas and proved to be a useful method for noninvasive follow-up in these lesions. MR is an excellent method in the evaluation of intramedullary hematomas. It may also be useful for demonstration of larger intradural AVM and of thrombus formation in AVM after surgery or embolization procedures. PMID- 2980611 TI - Magnetic resonance imaging of spinal cord hemangioblastoma. AB - We retrospectively reviewed the magnetic resonance (MR) studies in nine patients with spinal cord hemangioblastoma. Five patients were studied with a low field magnet (0.35 T/0.5 T) and four with a high field magnet (1.5 T). Six tumors were thoracic and three cervical in location. Magnetic resonance imaging (MRI) was positive in all the nine patients. MRI permitted a specific diagnosis prior to angiographic evaluation of the cord in three patients. The most characteristic findings in such tumors were: an intramedullary vascular nodule, enlarged drainage vessels, diffuse enlargement of the cord, and/or intramedullary cyst. The high field magnet showed a greater sensitivity in detecting vascularity within the nodule and draining vessels than did the low field magnet. MRI should be the first procedure in patients with myelopathy and subarachnoid bleed of spinal origin and for screening patients with von Hippel-Lindau disease. PMID- 2980613 TI - Value of magnetic resonance imaging in the evaluation of patients with complete and high degree block due to intracanal neoplasm. AB - Magnetic resonance imaging (MRI) of 26 patients with high degree or complete epidural and/or intradural myelographic block, due to primary or secondary neoplasm were analysed. The ratio of false negative MRI was 11.5 per cent. MRI findings were considered to be indicative of lesser block than indicated by myelography in almost 50 per cent of patients. Correlation between the two modalities was highest in cases associated with significant vertebral collapse, malalignment and angulation of the spine and angulation of the spinal cord. PMID- 2980612 TI - Computed tomography and magnetic resonance imaging of intramedullary spinal cord tumors. AB - Magnetic resonance imaging (MRI) was found useful in the evaluation and management of patients with intramedullary tumors. Both T1 and T2 weighted images are mandatory for depicting intramedullary pathology and sizable change of the spinal cord. Intratumoral cysts and syrinxes were often associated with tumors and shown as a low signal intensity on the T1 weighted image. Computed tomography (CT) with intravenous iodine showed enhancement of tumor nodules, in all mixed gliomas, ependymomas and hemangioblastomas. Enhanced MRI using gadolinium-DTPA is equal or superior to enhanced CT in localizing intramedullary tumors. PMID- 2980614 TI - Magnetic resonance imaging of the cranio-cervical junction. AB - One hundred and thirty-three patients with a clinically suggested lesion of the cranio-cervical junction were investigated by magnetic resonance imaging (MRI). Based on pathologic findings in 68 cases, MRI is considered the diagnostic method of choice if a lesion of the cranio-cervical junction is expected. With this method it is possible to determine space-occupying lesions, and inflammatory, demyelinizing and degenerative changes. Only rarely contrast media are necessary in order to differentiate between tumor and tumor-edema. PMID- 2980615 TI - Magnetic resonance imaging in the diagnosis of the posterior fossa and the spinal column. AB - Magnetic resonance imaging (MRI) is more advantageous for evaluation of the posterior fossa and spinal column because it can delineate the brain stem and the spinal cord in multidimensional planes without intrathecal contrast and ionizing radiation. In 18 cases with posterior fossa tumors, MRI was definitely superior to computed tomography (CT) in 11 cases (61%) and was equally diagnostic in 5 cases (28%). In investigations in 58 cases of spinal disorders examined by MRI, MRI was useful in diagnosing intramedullary cord tumors, syringomyelias, herniated discs, and infections, but CT was still more diagnostic in spondylosis, ossification of the posterior longitudinal ligament, and canal stenosis. PMID- 2980616 TI - Results of the international extracranial/intracranial bypass study. Implications for neuroradiologists. AB - A randomized trial to determine whether bypass surgery would benefit patients with appropriate atherosclerotic lesions followed 1377 patients for almost five years, and found no benefit from surgery. This negative result has a number of potential implications for neuroradiologists including the expectation for other randomized trials to be carried out, such as for carotid endarterectomy, and the need for neuroradiologists to carry out trials for embolization of cerebral arteriovenous malformations and angioplasty of stenotic cervical or cranial vessels. We are reminded that, ultimately, the clinical outcome is the real measure of procedures, not pretty pictures. PMID- 2980617 TI - Intracerebral cavernous angiomas in magnetic resonance imaging. AB - The findings obtained by magnetic resonance imaging (MRI) in 8 cases of surgically and histologically confirmed intracerebral cavernomas are reported. Six of the malformations were located temporally, one was located on the floor of the fourth ventricle, and one in the parietal lobe. All of the 8 cavernomas could be clearly demarcated, both in the T2 weighted image and in the spin density image. They are demonstrated as inhomogeneous zones with high or no signals. The tumors had irregular contours, which were clearly set off from the surrounding parenchyma. In 2 cases, the hemorrhage later detected surgically could already be assumed from the MR image. In 3 cases, zones with weak signals were found in the tumor, which were considered to be calcifications. One case of a vein with laminar flow could be established. PMID- 2980618 TI - Center differences in a study of the efficacy of B-scan ultrasound. AB - As part of a four year NIH-sponsored multi-center trial to evaluate the efficacy of B-mode ultrasound imaging of carotid atherosclerosis, multiple readings of B scan and angiographic studies of 234 vessels were obtained to permit intramodality assessment of within-reader and between-reader variation. All readers were obligated to adhere to a common protocol that was monitored both onsite and at a central coordinating facility. Among the five participating centers, the within-reader correlation coefficients for measurement of minimal residual lumen ranged from 0.85 to 0.99 for angiography and 0.62 to 0.93 for B scan. Between-reader correlation coefficients for minimal residual lumen varied from 0.50 to 0.98 for angiography and 0.59 to 0.87 for B-scan. Analysis of center differences for other measurements demonstrated similar variations. This suggests that center differences may have a significant impact upon results of multi center trials even when identical protocols are utilized. PMID- 2980619 TI - Magnetic resonance imaging of cerebral cavernous angiomas. AB - Magnetic resonance imaging (MRI) proved to be far superior to computed tomography and angiography in detecting cavernous angiomas in 55 patients, with a total of 72 lesions. A cavernous angioma usually appears as a nodular lesion with a relatively short T1 and long T2, circumscribed by a hypointense ring. The MR signal varies according to histopathologic components. Usually, no significant mass effect is detected. Sometimes a rim of edema or gliosis is present (long T2). When a massive bleeding is present, differential diagnosis from chronic hematoma of other origin is not always easy, but may be possible from the clinical presentation and some MR findings. PMID- 2980620 TI - Classification of brain arteriovenous malformation nidus by magnetic resonance imaging. AB - T1 and T2 weighted high field magnetic resonance imaging (MRI), computed tomography (CT) and intraarterial digital subtraction angiography (DSA) were performed in 30 patients with brain arteriovenous malformations. T2 weighted MR sequences proved superior to both CT and intraarterial DSA for displaying the configuration of the nidus, its intrinsic morphology and the state of the surrounding parenchyma. As was evidenced by histologic examination of the nidus in 18 cases, high-signal intensity of the intervening or surrounding parenchyma on T2 weighted images indicates gliosis. Preliminary experience with 5 cases suggests that gliosis of the intervening or surrounding brain parenchyma may make it safer to attempt embolization in these patients. Based on these parameters, we introduce a classification of brain AVM nidus. PMID- 2980621 TI - High resolution CT angiography in the direct diagnosis of cerebral aneurysms. AB - By performing a rapid series of 1.5 mm slices, with high resolution CT angiography (angio-CT), after intravenous bolus injection of 100 ml of contrast medium, an image of the basal arteries similar to that of conventional angiography may be obtained. A consecutive series of 102 patients with suggested cerebral aneurysms were examined by angio-CT and, as a control, were also studied with selective four-vessel angiography and/or at autopsy. In 33 patients angiography or autopsy revealed no aneurysm; 69 patients were shown to have 76 aneurysms. The angio-CT was correctly negative in 30 patients (91%), falsely suggested in 2 (6%), and falsely positive in 1 patient (3%). Sixty-eight aneurysms (89%) of down to 3 mm in size were correctly demonstrated by angio-CT, 6 (8%) were correctly suggested, and 2 aneurysms (3%) escaped CT demonstration. PMID- 2980622 TI - Computed tomography, dynamic cerebral scintigraphy and angiography in the diagnosis of cerebral aneurysms and ensuing blood flow disorders. A correlative study. AB - Twenty-nine consecutive patients were examined 3 to 6 days after subarachnoid hemorrhage, using conventional computed tomography (CT), computed angiotomography (angio-CT), conventional four-vessel angiography, and, in 20 cases, dynamic cerebral scintigraphy (DCS). Angio-CT indicated aneurysms in 21 cases. There was also agreement between angio-CT and angiography as to the localization of the aneurysms. DCS demonstrated blood flow disturbances in 15 cases with or without aneurysms. As opposed to these findings, changes due to blood flow disturbance were observed only in 7 cases at CT. The results suggest that angio-CT is a valuable method for detecting cerebral aneurysms and may limit the use of conventional angiography. DCS is a more sensitive method than CT, but DCS changes do not permit recognition of the type and location of the pathologic changes. PMID- 2980623 TI - A non-invasive Doppler ultrasound method for the evaluation of patients with subarachnoid hemorrhage. AB - Measurements of flow velocity in the basal cerebral arteries can be obtained non invasively through the intact skull using 2 MHz pulsed Doppler ultrasound. We investigated 47 patients with subarachnoid hemorrhage and compared flow velocity with angiography. Flow velocity in the middle and posterior cerebral arteries (MCA and PCA) was inversely related to the lumen diameter. Further clues to diagnose MCA spasm were obtained from the ratio of MCA flow velocity divided by the velocity in the distal extracranial internal carotid artery. In the individual patient, the two anterior cerebral arteries should be considered together, since this communicating system often has unique possibilities for collateral flow. Recordings from large MCA aneurysms were obtained before angiography in 4 patients. PMID- 2980624 TI - Transcranial Doppler sonography. An alternative to angiography in the evaluation of vasospasm after subarachnoid hemorrhage. AB - With transcranial Doppler sonography (TCD) the blood flow velocity (FV) in the basal cerebral arteries can be measured non-invasively. Vasospasm produces an increase of the FV in the narrowed arterial segment which can be evaluated by TCD. A clear correlation between the diameter of the artery and the increase in FV could be demonstrated in the middle cerebral artery. Vasospasm can be detected by TCD non-invasively and angiography can be avoided during the critical phase. The time course of vasospasm can be followed by TCD instead of repeated angiographies, enabling an optimal timing of aneurysm surgery. PMID- 2980626 TI - How nurses use curriculum concepts. AB - A study conducted in 1985 and 1986 attempted to discover if a curriculum based on a conceptual framework results in students using the concepts in a way that indicates they are of value. Results showed that students were able to recognize and articulate some of the concepts examined in the study, but there was no evidence that this ability was transferred to problem-solving activity in either simulation or clinical practice. The central issue to emerge was the lack of predictability of student performance on tasks requiring use of the concepts examined. Further questions were raised regarding the differential performance of students identified as belonging to minority groups. PMID- 2980625 TI - Whole or parts--a theological perspective on 'person'. AB - The influence of a theological perspective on nursing may be seen in the ethical debates concerning the beginning and end of life, the presence of theologians on ethics committees demonstrating the scope of opinion being sought. The meeting of theology and nursing to be addressed in this paper centres on the interaction of one human being with another, characterized by the terms community, relationship, response. Theology is concerned with persons reaching their potential not in isolation from, but in community with others. Fragmented care ensues from dividing a person into 'manageable' parts. Christian theology knows no such separation and offers a focus for an understanding of 'person' which has relevance for holistic nursing. The same holds true for groups as well as individuals; to focus primarily on self-interest is to ignore the possibilities for mutual exchange. Living 'in relationship' is both the point of departure and the ultimate goal of all created existence. Autonomy and mutuality are two sides of the same coin and the pursuit of one at the expense of the other is, according to Christian theology, not only illusory but a denial of the purpose of creation. PMID- 2980627 TI - Education for professional socialization in nursing. AB - The professional socialization of nursing students is seen as a developmental process occurring primarily during the period of formal education. As such, it cannot be left to chance but should be consciously considered in all generic nursing programs. PMID- 2980628 TI - Quality versus quantity of life: who should decide? AB - The issue of who should rightly decide quality versus quantity of life questions in health care contexts continues to receive widespread attention by health care professionals, academics and the laity. Unfortunately it is far from being resolved. Doctors still consider themselves the rightful, dominant decision makers. There are many cases where patients' lives have been maintained by extraordinary means without their informed consent, sometimes against their explicit will. However as a better informed public begins to assert its rights, the doctor's role as primary decision-maker in health care is being increasingly questioned. PMID- 2980629 TI - Effects of admission to a nursing unit. AB - This paper describes the results of a study of elderly patients admitted to hospital with fractured neck of femur, cerebral vascular accident or amputation of a lower limb who were, immediately post-crisis, transferred to a unit where nursing was perceived as the chief therapeutic activity. Patients' outcomes were compared with a control group who followed a normal patient pathway, either remaining in an acute hospital ward until discharge or being transferred to a community hospital for part of their stay. The study was a modified replication of a small pilot study carried out between 1983 and 1985. PMID- 2980630 TI - Advocacy and nursing: implications for women's health care. AB - A recent government report indicates that women in Victoria are dissatisfied with the health care they receive. Advocacy is presented as a core concept which could be utilized by nurses to effect change in women's health care. Some models of advocacy in the nursing literature are examined and related to the aims of the women's health movement. The model of patients' rights advocate appears to have the greatest congruence with the aims of this movement and could be adopted by nurses to improve women's experience of the health care they receive. PMID- 2980632 TI - Five days: five nursing theories. AB - Those who implement nursing theory presume that the patient will benefit, but from the patient's perspective, is any difference experienced as a result of the staff's deliberate adoption of a specific nursing theory? This paper is an imaginative attempt to look at nursing theory from the patient's viewpoint. PMID- 2980633 TI - Application of Orem's self-care model to nursing practice in developmental disability. AB - Orem's self-care model of nursing is analysed and applied to nursing practice within developmental disability services. A high degree of compatibility is found between Orem's model and current philosophies and practices in the field. Particular attention is focused on the relationship between Orem's concept of normalcy and the principle of 'normalization' which acts as the foundation of most modern services for people with developmental disabilities. A discussion of Orem's nursing systems and their interpretation in terms of the 'least restrictive alternative' is also given. PMID- 2980631 TI - A study of nurse leadership. AB - Little research has been undertaken on the leadership effectiveness of head (charge) nurses or on the preparation needed for this position. This descriptive correlational study explored the relationships between leadership effectiveness of head nurses and several educational and career variables. The study, which was undertaken in the northwest of USA, should be relevant to Australian nurses. Findings indicated that previous leadership experience (in school, college, community or professional committees) and nursing management experience prior to assuming the present head nurse position were positively related to the level of leadership effectiveness. A negative relationship was found between the length of time in the present position and the level of leadership effectiveness. The best combination of variables for predicting leadership effectiveness of head nurses, in order of the proportion of contribution, was found to be: previous nursing management experience; leadership continuing education; length of time in the present position; previous leadership experience and level of nursing education. These variables were found to positively influence leadership effectiveness, with the exception of length of time in the present position which was found to have a negative influence. PMID- 2980634 TI - Paradoxical priorities in breastfeeding research: challenges for new directions. AB - In recent years there has been a greater amount of research on infant feeding by medical and nutritional scientists than by behavioural and social scientists. The consequences of this imbalance of research focus is an extensive amount of information about breast milk with a lack of knowledge about breastfeeding. If we wish to increase the rate and duration of breastfeeding, greater attention needs to be paid to the art of nursing by resetting research priorities to take account of the mother's perspective. PMID- 2980635 TI - Nursing diagnosis: educational workshops to promote skill and inter-rater reliability. AB - Educational workshops focusing on developing skill in use of nursing diagnostic terminology in the workplace were provided over a 12 week period for a sample population of registered nurses (RNs) in an acute care facility. Inherent in the workshop format was the concept of inter-rater reliability, analysis and resolution of differences. Following participation in the workshops, pairs of RNs conducted assessments on selected stable patients in the agency and completed a nursing diagnosis data collection tool for each. Data were analyzed to determine inter-rater occurrence agreement and total agreement. Most diagnostic category disagreement was related to differences in observations or missed observations. Disagreement over etiologies and defining characteristics for agreed diagnostic categories was rare. In general there was a paucity of defining characteristics for most diagnostic categories generated. Future workshops must emphasize the necessity of a valid database prior to diagnostic formulation. PMID- 2980636 TI - Generating alternatives in nursing: turning curriculum into a living process. AB - The transfer of nursing education into higher education institutions is challenging nurses to re-think educational structures and processes. Traditionally, nursing education has used objective curriculum models which tend to reinforce bureaucratic rather than professional values. Critics suggest that these models, by their failure to incorporate social, political and cultural realities, contribute to theory-practice dichotomies. This paper shares the intentions of Deakin University's School of Nursing in developing alternative approaches to curriculum design and to learning and practising nursing. PMID- 2980637 TI - Calculating nursing turnover in NSW. AB - A study of nursing staff turnover is being carried out by the Nursing Research Team from the Riverina-Murray Institute of Higher Education for the New South Wales College of Nursing and the New South Wales Department of Health. The general aim of the study is to provide information about the annual rate of loss of registered nurses from public hospitals and licensed private hospitals in NSW. This paper provides background details about the project and recent attempts to understand the problem of nursing staff turnover. Brief mention is made of Australian and overseas literature and recent calls for methodological and conceptual precision in calculation nursing turnover. Reference is made to 'Nursing Turnover Indices' and other statistical descriptors used to guide this research. PMID- 2980638 TI - The Delphi technique. AB - The Delphi technique is an easy-to-use research instrument which has become an accepted method of achieving consensus among individuals and experts. This paper describes its techniques, parameters for its successful application, variety of uses and some advantages and disadvantages. PMID- 2980639 TI - Teleconferencing: peer support for new graduates and preceptors. AB - Teleconferencing was trialed as a means of providing peer support to preceptors and graduates participating in a graduate transition program. The project involved 35 registered nurses from eight different hospitals within New South Wales and this report summarizes participant evaluation of the first seven teleconferences. Despite some problems making telephone contact with on-duty nurses, 96 percent of respondents agreed or strongly agreed that teleconferences provide the opportunity to contact other nurses and share experiences and 100 percent agreed or strongly agreed that they would like to be included in future teleconferences. While giving valuable support to preceptors and new graduates, teleconferencing may also be an effective means of offering peer support and/or continuing education opportunities to registered nurses working in isolated settings. PMID- 2980640 TI - Occupational exposure to chemicals and the offspring: conflicts of interest? PMID- 2980641 TI - Occupational diseases in Poland. Legal status and epidemiological situation. PMID- 2980642 TI - Fibrogenic and carcinogenic effects of antigorite. PMID- 2980643 TI - Mortality among female workers in an asbestos factory in Poland. PMID- 2980644 TI - Mutagenicity testing of air pollutions in a non-ferrous metal foundry. PMID- 2980645 TI - The impact of EMULKOP on the biological aggressiveness of coal mine dusts. PMID- 2980646 TI - Sickness and accident-related absenteeism in Poland between 1984 and 1986. PMID- 2980647 TI - High-frequency (10-20 kHz) hearing threshold in 20-year-old subjects previously unexposed to noise. PMID- 2980648 TI - Magnetic field intensity and energy absorption due to short-wave induction diathermy. PMID- 2980649 TI - [Life support care in hopeless cases]. PMID- 2980650 TI - [Effect of low-protein diet on the control of progression of chronic renal failure]. PMID- 2980651 TI - [Clonal growth of erythroid progenitor cells from normal bone marrow exposed to leukocytes from the blood of patients with acute leukemia]. PMID- 2980652 TI - [Studies of stimulating and conducting activities of the heart in hyperthyroidism]. PMID- 2980653 TI - [Effect of cimetidine on radioisotope-labeled thyroxine absorption from the digestive tract]. PMID- 2980654 TI - [Late results of endoscopic treatment of large-intestinal adenoma]. PMID- 2980655 TI - [Study of disorders of the ventilatory function of the lungs based on serial spirometric examinations of patients before and after surgical treatment of mitral valve defects. I. Preoperative studies]. PMID- 2980656 TI - [Study of disorders of the respiratory function of the lungs based on serial spirometric examinations of patients before and after surgical treatment. II. Control studies]. PMID- 2980657 TI - [Plasma vasopressin changes in patients with renal failure during water immersion]. PMID- 2980658 TI - [Sodium, potassium, ATP, 2,3-diphosphoglycerate and inorganic phosphate levels in erythrocytes of patients on hemodialysis]. PMID- 2980659 TI - [Effect of selective adrenergic stimulation on metabolic reactions in obese and slender women]. PMID- 2980660 TI - [Circulating immune complexes in idiopathic and secondary thrombo- cytopenia. II. Comparison with acute phase factors]. PMID- 2980661 TI - [Aluminum in human pathology]. PMID- 2980662 TI - [Reminiscence on Prof. Zbigniew Latallo (1924-1987)]. PMID- 2980663 TI - The effects of nutrition on brain function, behavior and learning: directions for integrative research. PMID- 2980664 TI - Limbic traces and interictal behaviour: basic science and clinical approaches. PMID- 2980665 TI - Complex behavioral chains of temporo-limbic epilepsy and their relationship to emotional phylogenesis: ictal laughter. PMID- 2980667 TI - Neural substrate for emotion: relationship of feelings, sensory perception, and memory. PMID- 2980666 TI - Sexually dimorphic pattern in the hypothalamic and limbic brain. PMID- 2980668 TI - Brain mechanisms and aggressive behavior in the cat. PMID- 2980669 TI - Some neuroendocrinological effects of socalled anxiolytic music. PMID- 2980670 TI - Emotion and immunity. PMID- 2980671 TI - Symptoms of limbic dysfunction in the acute psychoses of Zimbabwe. PMID- 2980672 TI - Modification of emotional expression induced by clinical and experimental epileptic disturbances. PMID- 2980673 TI - The physiological link between epilepsy and psychosis. PMID- 2980674 TI - Development and differentiation of the sexually dimorphic nucleus in the preoptic area of the rat brain is under hormonal control. PMID- 2980675 TI - Suppression of seizures and psychosis of limbic system origin by chronic stimulation of anterior nucleus of the thalamus. PMID- 2980676 TI - History of medicine. PMID- 2980677 TI - Affective symptomatology in epilepsy. PMID- 2980678 TI - Midbrain neural mechanisms mediating emotional behaviour. PMID- 2980679 TI - Effects of temporal lobe epileptiform activity upon aggressive behavior in the cat. PMID- 2980680 TI - Emotional disturbances in limbic system dysfunction. PMID- 2980682 TI - Memory. XIIth International Symposium of the Fulton Society. San Francisco. PMID- 2980681 TI - Hypothalamic output controlling reticulospinal and vestibulospinal systems important for emotional behavior. PMID- 2980684 TI - Neurological and behavioral investigations of memory failure in aging animals. AB - Aged organisms show a decline in their ability to learn and remember in certain situations. For example, it appears that both aged humans and aged rats exhibit spatial memory deficits. It is more difficult for older organisms to learn about and to navigate accurately within a familiar environment than it is for younger organisms. Because the brain structure that is critically involved in this type of behavior (the hippocampus) is relatively well understood in the rat, a correlation of spatial behavior with hippocampal physiology has been particularly useful in the delineation of some of the potential brain changes responsible for memory changes with age. Evidence for an age-related deficit in spatial memory is presented that emphasizes the importance of the contribution of the spatial component to the learning/memory changes seen with age in rats. The contribution of changes in hippocampal synaptic plasticity and in the information processing characteristics of single hippocampal cells of old rats is also discussed in terms of the potential influence on old animals' performance on spatial tasks. PMID- 2980683 TI - Can memories be encoded across generations? AB - Studies of familial psychopathology, specifically of "imperative ideas and actions", indicate a frequent transgenerational occurrence. These affectively highly charged complexes (memories) are encoded, it is speculated, across generations by a genetic mechanism. PMID- 2980685 TI - Mechanisms of behaviorally-elicited and electrically-elicited long-term potentiation. AB - Electrically-elicited LTP in the hippocampal formation need not depend for either elicitation or its maintenance, upon any region of the brain outside the hippocampal formation. Behaviorally-elicited LTP certainly must depend for its elicitation upon input from other brain regions, specially the perforant pathway from the entorhinal cortex showa potentiation. Certainly there is variation in the evidence that spine shape change occurs in response to behavioral situations. Lee et al and Chang and Greenough have reported increased synaptic density, indicating the formation of new synapsee in subfield CA1 of the hippocampus following the induction of LTP. Synapses may form in responses to behaviorally induced neural activity. Synapse formation is also compatible with other physiological changes reported to be associated with LTP induction such as increased neurotransmitter release and increased receptor numbers. Thus it remain perhaps the most likely basis for LTP and one of the most likely candidates for involvement in long-term memory in general. PMID- 2980686 TI - Anatomical substrates of interictal memory deficits in temporal lobe epileptics. AB - Temporal lobe epileptics frequently have as their pathological substrate selective neuron loss of the hippocampus. The extent and pattern of the hippocampal cell loss vary among patients. These patients also exhibit varying degrees of memory loss. We have demonstrated a relationship between the memory loss in left temporal lobe epileptics and their hippocampal lesions. Specifically, we have identified a memory measure, delayed-recall of unrelated word-pairs, that is highly dependent upon integrity of left hippocampus. We have also shown that extensive damage throughout the hippocampus is necessary to consistently impair this measure. These findings are relevant to understanding the memory loss of temporal lobe epileptics as well as providing insight into our understanding of the anatomical basis of memory. PMID- 2980687 TI - Associative learning and long-term potentiation: cellular mechanisms compared. AB - The potential relationship of LTP (Long Term Potentiation) to behavioral learning is an important issue. An important question in whether the cellular mechanisms of LTP and naturally occurring learning, share the same "memory" formation process or whether these two phenomena are subserved by two different types of storage processes. We would suggest that at least in the case of the hippocampal region, LTP could well share common mechanisms of formation with well studied examples of both cognitive and non-cognitive, or habit memory. We have considered several lines of evidence which demonstrate striking similarities between the phenomena of learning, especially associative learning and LTP. PMID- 2980688 TI - Further evidence in support of the neurobiological bases of an attribute model of memory: role of the hippocampus. AB - With respect to the nature of information processing within the hippocampus, it can be concluded that the hippocampus codes spatial attributes, may or may not code sensory-perceptual attributes, appears to code temporal attributes based on spatial and sensory-perceptual cues, but does not code response attributes. The data provide some support for each theoretical view of hippocampal function, but appear to be most consistent with Kesner's views. With respect to how information is coded within the hippocampus, it can be concluded that there is good support for hippocampal involvement in a working or data-based memory system, but not a reference or expectancy-based memory system. These data support the Olton and Kesner views of hippocampal function. With respect to dynamic processes associated with maintaining information within the hippocampus, it appears that spatio-temporal information is processed in the hippocampus across the whole time domain including both short term and intermediate term memory. Even though some information appears to be processed only in an intermediate term memory (Rawlins, 1985), it seems that at least for spatio-temporal information the apparent residual short term memory capacity is a function of remaining hippocampal tissue. PMID- 2980689 TI - Cortical and subcortical structures involved in recognition memory: neurophysiological and anatomical studies. AB - Recordings of neuronal activity were made in cortical and subcortical structures of monkeys performing visual recognition memory tasks in which the novelty/familiarity of the stimuli was varied. A population of neurons with memory-like activity was recorded in a periventricular region at the border of the anterior thalamus and dorsal hypothalamus. These neurons responded with decreases in firing rate to novel stimuli and increases in firing rate to the same stimuli when they were familiar. The calculated memory spans of these neurons ranged from 7 to hundreds of intervening trials. The afferent connections of this region were traced by an injection of HRP into the periventricular region. Retrogradely labeled cells were observed in the ventromedial prefrontal cortex, the ventromedial temporal cortex, amygdala, medial thalamus and supramammillary region. Recordings were then made in the hippocampus and ventromedial temporal cortex. A population of neurons in the ventromedial temporal cortex responded maximally to the first presentation of a stimulus, reflecting the novelty or recency of the stimulus presentation. In contrast, such neurons were not observed in the CA fields or subiculum of the hippocampal formation. These studies suggest that a system of brain structures involving the temporal cortex and periventricular region provide one mechanism whereby monkeys are able to recognise and respond to visual stimuli differing in their novelty and familiarity. PMID- 2980690 TI - The organization and neural substrates of human memory. AB - The neurology of memory has been illuminated by parallel studies of patients with circumscribed memory impairment and animal models of human amnesia. Human amnesia can occur as an isolated cognitive deficit that impairs the ability to learn new facts and episodes. In addition, memory can be affected for material learned many years prior to the onset of amnesia. The finding that some memory abilities are intact in amnesia (e.g., skill learning, word priming, and adaptation-level effects) has suggested that memory can be divided into two or more separate processes. Declarative memory affords the ability to store information explicitly and to retrieve it later as a conscious recollection. This form of memory depends on the integrity of the structures damaged in amnesia. Other, non-declarative kinds of memory afford the ability to change as the result of experience, but the information is available only through performance. Recent studies of a favorable human case provided strong evidence that the hippocampus is a critical component of the declarative memory system. Extensive convergent and divergent projections link the hippocampus to many areas of neocortex where processing and storage of new information is likely to occur. It is perhaps by way of these connections that the hippocampus operates upon and participates in declarative representations. PMID- 2980691 TI - Memory traces: how to increase and decrease their strength. AB - In addition to the memory trace itself, the memory circuit may include a modulatory input that influences the strength of the memory trace. This modulatory input, which can make memory traces stronger as well as weaker, appears to be intimately associated with systems that convey the importance of the present event. A single exposure to a sufficiently salient event may activate the modulatory input to a level high enough to produce an enduring memory. Experimental investigation of this modulatory input indicates that pharmacological treatment can often be substituted for practice to strengthen the memory trace, suggesting that drugs or practice activate a common mechanism. Similarly, drugs and the UCS interact in such a way as to suggest they also act on a common mechanism to modulate retention. Dose-response curves for drug treatment or UCS amplitude versus performance are often U-shaped. Many drugs that modulate memory appear to do so through a primary site of action outside the blood-brain barrier. Our investigation of one such peripheral system, namely that for [leu]enkephalin, revealed an enzyme mechanism in plasma which may be regulatory with regard to escape performance. We suggest that investigation of the interface between the memory trace and its modulatory input may lead to understanding of how mammalian systems store information, particularly in regard to associative strength. PMID- 2980692 TI - The irrelevance of mammillary body lesions in the causation of the Korsakoff amnesic state. AB - This discussion has been concerned with one small corner of the anatomy of memory, namely, with the role of mammillary body lesions in the causation of the Korsakoff amnesic state. Clinical-pathologic studies, both our own (of alcoholics with the Wernicke-Korsakoff syndrome) and those of others (in patients with thalamic infarctions and neoplasms) provide unambiguous evidence that medial thalamic lesions, without additional lesions of the mammillary bodies, are quite sufficient to produce a severe and enduring amnesic state. Behavioral abnormalities in animals, insofar as they can be equated with a defect in retentive memory, tend to substantiate the observations in humans. PMID- 2980693 TI - Age-associated memory impairment: diagnostic criteria and treatment strategies. AB - In general, then, the term Age-Associated Memory Impairment (AAMI) has been adopted to describe healthy persons over 50 years of age who have experienced memory loss since early life, but who are not demented. Specific diagnostic criteria have been proposed for AAMI and these criteria are now being applied in clinical studies, including a number of pharmacologic treatment studies. Many of these latter studies are supported by evidence from preclinical studies suggesting that age-related memory deficits in animals can be pharmacologically altered. This evidence gives reason to hope that an important behavioral deficit associated with aging may be controlled or corrected through drug treatment. The development of such a treatment would be an accomplishment of immense clinical and scientific significance. PMID- 2980694 TI - Predictive regulation of associative learning in a neural network by reinforcement and attentive feedback. AB - At least four types of learning processes are relevant in the present paper: learning of conditioned reinforcement, incentive motivation, sensory expectancy, and motor command. These several types of learning processes, which operate on a slow time scale, regulate and are regulated by rapidly fluctuating limited capacity STM representations of sensory events. The theory suggest how nonlinear feedback interactions among these fast information processing mechanisms and slow learning mechanisms participate in different conditioning paradigms, and actively regulate learning and memory to generate predictive internal representations of external environmental contingencies. PMID- 2980697 TI - Phase-contrast microscopy of the primate retina. AB - The study of hematoxylin and eosin stained thick sections (15 microns) of the primate retina with the phase-contrast microscope provided a means for the selective demonstration of many cellular structures that could not be resolved with the same degree of detail which was possible when bright-field microscopy was used, or when phase-contrast microscopy was employed to examine unstained material. The H & E-stain greatly enhanced the phase-contrast image, so that cytoplasmic structure, fiber trajectories, and gross synaptic detail of the retina could be demonstrated to better advantage. PMID- 2980696 TI - The Haitian Health Foundation (formerly the Haitian Dental Foundation)--a charitable outreach to neighbors in need. PMID- 2980695 TI - Neuropeptides, environmental and social approach behaviour in the investigation of emotion. PMID- 2980698 TI - Immunofluorescent examination of the kidney post mortem. AB - 106 selected kidneys removed at autopsy were studied by direct immunofluorescence using polyvalent antisera against human immunoglobulins, light chains, complement fractions and fibrinogen. The immunofluorescence was a suitable method to solve differential diagnostic problems that arose at autopsy. The diagnostic value was the most obvious in cases of immunologically mediated renal diseases and in immunologically mediated systemic diseases involving the kidneys. Negative immunofluorescence findings were also useful to determine the pathogenesis of renal lesions, especially in vasculopathies. The immunofluorescence of postmortem material showed similar disturbances to that obtained with biopsy material. At various sites, especially in the tubulo-interstitium, additional electron microscopical study was sometimes needed to localise the immune deposits exactly. The fluorescent microscopical examination of frozen sections of kidney taken at necropsy turned out to be more adequate than the immunoperoxidase examination of formalin-fixed, paraffin-embedded sections. PMID- 2980699 TI - The ultrastructure of alcoholic liver disease: a review and analysis of 100 biopsies. AB - Liver biopsies from 100 patients with alcoholic liver disease of various grades of severity were examined by light and electron microscopy. A comprehensive account of ultrastructural morphology is presented. The organellar changes were variable both in nature and intensity. The most consistent ultrastructural changes, irrespective of disease severity, were dilatation of endoplasmic reticulum and fat accumulation. The former is the EM counterpart of hepatocyte swelling at the light microscope level. Collagen deposition was detected earlier and more accurately by electron microscopy. A physical relationship between Mallory bodies and intermediate filament was also detected. As is the case with light microscopy, if cellular, stromal and organellar changes are considered independently, some may be regarded as typical, but none as pathognomonic of alcoholic liver disease. PMID- 2980700 TI - Muscle fiber types in tetrapods. A comparative histochemical and morphometric study. AB - A comparative histochemical and morphometric study in two groups of homologous muscles from different tetrapods (rat, pigeon, lizard and frog) was performed. On the basis of their fiber diameters and oxidative enzyme activities, an initial correlation between fiber types of all animals is observed, although in the lizard and frog muscles, another fiber type does exists that could not be demonstrated in higher vertebrates. When more than one histochemical techniques are used for the identification of each tetrapod fiber types, the lack of correlation between them becomes obvious. Thus, different animals groups, each showing a characteristic muscle metabolic pattern, could be distinguished. PMID- 2980701 TI - Ultrastructure of the area postrema of the monkey, Macaca fascicularis. AB - The area postrema of the monkey, Macaca fascicularis, were a pair of oval organs at the caudal end of the floor of fourth ventricle. Their ependymal lining was covered by well-developed microvilli with occasional overlying supraependymal cells. Two types of lining cells were present: pyramidad- and flattened cells. The pyramidal cell showed a long extending basal process resting on the underlying blood vessels. In transmission electron microscopy, the organ showed numerous fenestrated sinusoids characterized by a distinct perivascular space containing mast cells, macrophages and collagen fibrils. The parenchyma of the organ was composed of neurons and glial elements. Only one type of neuron ranging from 9.5 to 15 microns could be distinguished. The neurons contained an indented nucleus surrounded by organelle rich cytoplasm. The soma of the neuron was enclosed by glial element resembling astrocyte. The glial processes terminated on the blood vessel where they were "tunnelled" by a variable number of nerve fibres some of which gained a direct access to the external basal lamina of the perivascular space. Synapses in the neuropil predominantly of the axodendritic variety were observed. Axon terminals containing round agranular vesicles were seen to make synaptic contacts with the neuronal soma. No structural changes were observed in the area postrema following bilateral cervical vagotomy. However, degenerating axon terminals were observed in the subpostremal zone 7, 14 and 21 days after vagotomy suggesting a direct afferent projection into this region. PMID- 2980703 TI - Purpura cerebri in gram-negative septicaemia. A histological and immunohistochemical study. AB - Two cases with brain purpura following Gram-negative septicaemia were examined morphologically and immunohistochemically. The brain lesions, including ball and ring haemorrhages, a few days old, with some microglial cells accumulated around the older foci, were restricted to the white matter. Immunohistochemically, scanty deposits of IgG, IgA and IgM mainly in the macrophages in brain, kidneys and lungs were found, whereas staining with antibodies directed against IgE and complement (C3, C4) remained negative. In the brain, immunoglobulin deposits were located mainly in the macrophages, furthermore, in and around the walls of a few intact (non-haemorrhagic) vessels; within the perivascular haemorrhagic foci no deposits could be demonstrated. The relevance of these observations to the pathogenesis of brain purpura is discussed. PMID- 2980702 TI - Hyaline-vascular type of Castleman's disease (angiofollicular lymph node hyperplasia) with monotypic plasma cells. An immunohistochemical study with monoclonal antibodies. AB - A case of angiofollicular lymph node hyperplasia (Castleman's disease) characterized by monotypic (IgG+, lambda+) plasmacytosis is described. Fresh tissue was available and a thorough immunohistochemical analysis of lymphoid and non-lymphoid cells was performed on cryostat sections. Although lymphoid follicles were numerous and exhibited some abnormal features they did not appear part of the monoclonal cell proliferation. Follicular lymphocytes were mixtures of Kappa+ and lambda+ cells. Vessels penetrating within these abnormal follicles expressed reduced levels of FVIII and Leu-M5 antigens and exhibited thicker layer of collagen type IV. The analysis of T-cell subsets showed a normal (3:1) T4/T8 ratio. This case extends to the mixed variant of hyaline-vascular Castleman's disease, the neoplastic potential previously associated to the plasma cell variant of the disease. PMID- 2980704 TI - Scanning electron microscopy of the aggregation of head mesoderm cells from chick embryos. AB - Head mesoderm cells from chick embryos at different stages of development were dissociated and cultured on plastic coverslips. In all cultures several cellular aggregates were described by means of scanning electron microscopy. Isolated cells present filopodia and lamellipodia. However, when mesoderm cells make contact with one another the filopodial and lamellipodial activity in the contact cellular edge disappear. Thus, the cells into cellular clusters do not present projections. The clusters were circular and bidimensional in character. The scanning electron microscopic observations showed that it is the type 1 variant of "contact inhibition of locomotion" which occurs. By means of these mechanisms the bidimensional aggregates are formed and cellular overlapping is not present. Since the behaviour of the mesoderm cells "in vitro" in some way could be comparable to their behaviour "in situ", the results here observed are discussed in relation to the conduct of mesoderm cells "in vivo". PMID- 2980705 TI - Spontaneous complete clefting of the palates in a mouse fetus: a study by scanning electron microscopy and serial section reconstruction. AB - A 15 day mouse fetus having spontaneous complete clefting of the primary and secondary palates was studied in comparison with its normal litter mates and with normal 14 day fetuses. Specimens were studied by scanning electron microscopy at various stages of microdissection, by light microscopy of thin serial sections and by serial section reconstruction of the anterior chondrocranium of the clefted specimen and one of its normal litter mates. Differentiation of tooth and bone tissue was slightly retarded in the clefted fetus but paranasal and oral landmarks, though distorted, were present. The clefted fetus had a smaller angle between cranial base and nasal capsule and a marked discontinuity between the primary and secondary palates. Cell surfaces on the medial edge of the secondary palate in the clefted fetus resembled cell surfaces of oral areas that do not normally fuse, i.e. they are polygonial, flat and bear few surface projections in contrast to the normal 14 day condition where these cells are spindle shaped, convex and have many microvilli. The observations support the concepts that clefting of the secondary palate is consequential to clefting of the primary palate, that maldevelopment of neural crest mesenchyme is not necessarily a contributing factor, that clefting of the primary and secondary palates is associated with a shorter anterior-posterior dimension of the head and that when fusion of palatal shelves fails to occur the cells of the medial edges modulate in the direction of a generalized type of surface epithelium. PMID- 2980706 TI - Electron microscopy and immunostaining of the normal breast and its benign lesions. A search for neuroendocrine cells. AB - Specimens from 7 patients with normal breast tissue 26 patients with benign breast lesions (6 fibroadenomas, and 4 intraductal papillomas, 2 mammae lactantes, 10 cases of cystic disease and 4 fibrotic lesions) were studied by immunocytochemistry and electron microscopy. Excretory epithelial cells in 2 of the 4 papillomas were immunostained for NSE. Myoepithelial cells were frequently stained as well. All the breast specimens were nonreactive to the antichromogranin antibody we used. The 2 NSE positive intraductal papillomas were tested for presence of hormone immunoreactivity, but no positively stained cells were observed. No cells with neuroendocrine features were observed by electron microscopy. The present study did not reveal neuroendocrine cells in the normal breast specimens and undisputed proof of neuroendocrine differentiation in benign breast lesions was not established. We conclude that if neuroendocrine cells are present in the normal breast, they are very rare, and probably not the cellular origin of all breast carcinomas with neuroendocrine features. PMID- 2980707 TI - The response of subpleural pulmonary capillary endothelium to hydrothorax in rats. AB - The principal focus of this study was to evaluate the hypothesis that increased interstitial fluid pressures served to stimulate de novo vesicle formation in pulmonary capillary endothelium. Direct measurements of interstitial fluid pressures within the alveolar septa pose great technical difficulty. The pleural space and subpleural capillaries are easily accessible, and thus, provide a more feasible model to test this hypothesis. After hydrostatic pressure of pleural space fluid was increased by periodic saline infusions into the pleural cavity, vesicle numerical densities were significantly increased in portions of the subpleural capillary endothelium. Those segments of the endothelium that directly apposed the interstitium of the visceral pleura displayed de novo vesicle formation. The endothelial segments located immediately adjacent to the alveolar epithelium were not affected by the elevated interstitial fluid pressures. In addition to the increased vesiculation, those same segments of the endothelium were characterized by increased attenuation of their cytoplasmic compartments. These conformational changes in the plasmalemma of portions of the subpleural capillary endothelium provide support to the tentative hypothesis, however, whether the increased numbers of vesicles contribute to a potential transendothelial transport system or expand a possible static network of membrane invaginations remains uncertain. PMID- 2980708 TI - Studies on intra-arterial cushions. IV. Perivascular nerve plexuses of ramifying arteries with intraluminal cushions at the branching points. AB - Periarterial nerve plexuses were studied at branching points of arteries with intraluminal cushions. Serial sections were prepared from mouse kidney, pancreas and tongue, and studied by means of catecholamine fluorescence and staining for acetylcholinesterase. The periarterial nerve plexuses did not show any peculiarities at the branching points. The intraluminal cushions, as well as the tunica media of both the parent trunk and the collateral branches, were found to be free of vegetative nerves. It is concluded that the shape of intra-arterial cushions is passively altered, following the alterations of vascular geometry, and not in response to direct nervous stimulation. PMID- 2980709 TI - Stereologic analysis of the in vivo alveolar type II cell response to isoproterenol or saline administration. AB - Previous studies have demonstrated enhanced secretion of pulmonary surfactant from type II alveolar epithelial cells following beta-adrenergic stimulation. The present study was undertaken in order to provide quantitative morphologic data supporting this effect in vivo. Adult male Sprague-Dawley rats were injected subcutaneously with 150 mg/kg L-isoproterenol, a wide-range beta-adrenergic agonist, and killed at times 0.25-12 hours post-injection. Other rats were similarly injected with saline, and killed at times 0.25-6 hours post-injection. A third group of animals was not injected, nor handled, prior to the time of death, and served as baseline controls. Stereologic analysis of the intracellular organelles of the type II cells in the animals treated with L-isoproterenol revealed a significant decrease in lamellar body volume density, indicating increased secretion of surfactant, at 0.5-4 hours. The rough endoplasmic reticulum volume density increased significantly at 2-6 hours, indicating increased synthetic activity. In contrast, the type II cells of saline-injected animals showed no significant evidence of increased secretion, but did demonstrate a large increase in synthetic activity, resulting in many large lamellar bodies at 2 and 4 hours post-injection. The results of this study provide quantitative morphological evidence of beta-adrenergic stimulation of the secretion and synthesis of pulmonary surfactant secretion by type II cells of the adult rat lung in vivo. In addition, they suggest an enhancement of surfactant synthesis following saline injection, which is perhaps based on endogenous catecholamine release. PMID- 2980711 TI - Endocrine cells of the gastric mucosa of Rana temporaria L. AB - The endocrine cells of the gastric mucosa of Rana temporaria have been studied according to the ultrastructure, the staining properties of the granules with Masson Fontana's and Grimelius' silver methods, silver impregnation of Davenport on deplasticised semithin sections and immunocytochemical techniques. Seven different types of endocrine cells have been described. Six were regarded as belonging to known types: G, A, EC, ECL, D and P cells. One type was considered as unclassifiable. PMID- 2980710 TI - Classification of routinely processed anaplastic large cell tumours with a small panel of antibodies. An immunohistochemical study with clinical follow-up. AB - A proportion of anaplastic large cell tumours is difficult to classify on sections of routinely processed, paraffin-embedded tissue. Differentiation into large cell lymphoma, carcinoma, melanoma or sarcoma is important in order to assess prognosis and proper treatment. Although the use of immunohistochemistry has been reported in the differentiation between some of these types of neoplasms, no antibody panel, which can directly differentiate all of them, has been described. In the present study we evaluated the value of a panel of 5 antibodies for the classification of 29 anaplastic large cell tumours, which could not be classified by experienced pathologists using conventional histological and histochemical techniques. The panel, which can be used on routinely fixed paraffin-embedded tissue, consisted of 5 different antibodies directed against keratin, vimentin, the human milk-fat globule membrane antigen MAM-6, a melanoma associated antigen and common leucocyte antigen. The use of this panel directly resulted in a definite diagnosis in 95% of the cases and provided valuable information for the diagnosis in the remaining cases. The diagnosis was confirmed by additional marker studies and electron microscopy. Moreover, clinical follow-up, including treatment data, was in accordance with the diagnosis based on the panel. PMID- 2980712 TI - Autoradiographic investigation of circadian rhythms in alveolar bone periosteum and cementum in young mice. AB - This report presents circadian rhythms in cell proliferation of alveolar bone periosteum and cementum of the maxillary first molars of male 5-week-old BNL, Swiss albino mice which were maintained on a 12 hr light/dark cycle. Mice were injected with 3H-TDR (luCi/gm. body wt.) 1 hr prior to sacrifice and killed every 3 hrs for 24 hrs starting at 9 a.m. Maxillae were decalcified, routinely processed histologically and autoradiographs prepared. Cell labeling indices of alveolar bone and cementum mesial to the first molar were determined. Alveolar bone periosteal and cemental cells show circadian rhythm in their DNA synthetic processes. Peaks in percent labeling exhibit higher values than previously reported for nontraumatized, normal dental periosteum and cementum. While the outer periosteum reveals a single 24 hr peak (6 p.m.), inner periosteum and cementum reveal two ultradian peaks 9 to 12 hrs apart involving both light and dark periods. Rodents are nocturnal, but high peaks are also evident in the light periods, consequently, not all peaks are synonymous with the period of animal activity and feeding. Although the single daylight peak of the outer periosteum may indicate growth of that surface at night to about noon, the double peaks exhibited by inner periosteum and cementum indicate light/dark, continuously active surfaces in terms of DNA synthesis and growth. PMID- 2980713 TI - Light microscopical morphometry of prolactin secreting adenomas under treatment with dopamine agonists. AB - In order to study the light microscopical alterations of pituitary tumours under dopamine agonist treatment, three groups of a total of 18 large or small cell chromophobe adenomas were analysed by light microscopical, immunohistological and morphometrical methods. They were all removed by transsphenoidal surgery. 6 of them were treated preoperatively with dopamine agonists, bromocriptine and/or lisuride, for various periods of time. 8 adenomas remained preoperatively untreated. 4 additional untreated tumors were small cell inactive adenomas for comparison. One case was excluded from the final evaluation of the data because it appeared to be a typical non-responder, clinically as well as histologically. Immunohistological positivity for prolactin was to be found in all cases in various degrees. Clinically active adenomas contained many prolactin positive cells, whereas in inactive adenomas only scattered cells were prolactin positive. The morphometric analysis revealed a reduction of the cytoplasmic area in a statistically significant degree in the group of adenomas under treatment, which explains adequately the shrinkage of the entire adenoma and the reduction of prolactin plasma levels. The morphometric data of treated adenomas resembled those of untreated inactive adenomas. PMID- 2980714 TI - Autoradiographic localization of estrogen target cells in the spinal cord of the armadillo and baboon: a comparative study. AB - The uptake and retention of radiolabeled estradiol by the spinal cord were examined in the baboon and the armadillo and compared to previous observations in the rat. Four females of each species were injected intracardially with 1.0-1.4 micrograms/kg body weight of 3H-estradiol and two females, one baboon and one armadillo, were injected with both labeled and 100-140 micrograms/kg body weight of unlabeled estradiol. One hour after the injections, the animals were killed and segments from the cervical, thoracic, lumbar and sacral cord were removed and processed for autoradiography. In the armadillo, labeling of neuronal nuclei were noted in laminae I & II and in alpha motor neurons. In addition, nuclei of the ependymal cells of the ventral portion of the central canal in the cervical cord concentrated radioactivity. In contrast, the baboon demonstrated only sporadic labeling of neurons in lamina II in all levels of the spinal cord. The comparison of our observations with that of the rat suggest that estrogen mediated sensations are probably coordinated at higher brain centers in the primate as opposed to the more primitive mammals. PMID- 2980715 TI - High-energy adhering junctional complexes or with mitochondrial coupling. AB - A variety of adhering junction is found in the ependyma of the domestic cat with a coupling of mitochondria. These are symmetrically situated (in mirror form) at both sides of the intercellular cleft, which always maintain the same separating distance, thereby leaving a limiting cellular space of a constant amplitude. The hypothesis is put forward that the energy (ATP), provided by the mitochondria over adhering junctional complexes, would produce separate fields of force which would position in a lengthwise direction the molecules which give rise to the anchoring filaments. The mitochondrial energy provided and the electrostatic forces generated would produce an adhering, intercellular junction which is functionally very strong and which could be called: high-energy adhering junctional complexes or with mitochondrial coupling. PMID- 2980716 TI - Immunocytochemical studies on prolactin cells in the adenohypophysis of the golden hamster. AB - Mammotrophs or prolactin (PRL) cells were identified in the adenohypophysis of adult golden hamsters by immunocytochemical techniques with a polyclonal anti PRL, that was proved to be specific to PRL by the dot immunoblotting test. Postembedding immunostaining was performed on Araldite thin sections by immunoperoxidase and immunogold methods. PRL cells were classified into three types according to the different size of the secretory granules. The Type A cells were usually small and angular or oval in shape, and had secretory granules ranging in diameter from 100-230 nm, and showed poorly developed organelles. The Type B and C cells were larger and round or ovoid in shape, contained larger granules, 230-280 nm and 280-570 nm, respectively, and displayed well developed organelles. Immunoreactive PRL cells in the male pituitaries were far less numerous than in the nonpregnant female glands, and were mostly of the Type A and B, whereas in the female the Type C and B cells predominated. In pregnant females, Type C cells became activated and increased in number, while the other two types decreased in proportion. In lactating females, Type A and B cells significantly increased in number at the expense of the Type C cells; meanwhile, the exocytosis of secretory granules was frequently found in all types of PRL cells. The present findings suggest that Type C and B PRL cells, especially the former, are potent in producing and releasing PRL and highly responsive to various physiological stimuli, while Type A cells are probably relatively inert in synthetic activity. PMID- 2980717 TI - The protective effects of a prostaglandin without antisecretory properties against ethanol-induced injury in the rat stomach: a histologic study. AB - This study examined the effect of 2-acetyl-2-decarboxy-15(S)-15 methyl PGF2 alpha (PGF2 alpha) on ethanol (EtOH) induced injury in the rat stomach to determine if a PG analogue devoid of antisecretory properties could confer full or partial gastric mucosal protection. Rats were orally administered saline or PGF2 alpha in a dose of 0.5 or 5.0 mg/Kg. Thirty minutes later animals received varying concentrations (i.e. 25%, 50%, and 100%) of EtOH orally. Five minutes following EtOH exposure, they were killed and samples taken from identical regions of the glandular mucosa for microscopic evaluation. All concentrations of EtOH tested damaged the gastric epithelium. The injury induced by 25% EtOH was almost exclusively confined to the surface epithelium and was not altered by either dose of PGF2 alpha pretreatment. In contrast, both 50% and 100% EtOH elicited comparable damage to the gastric mucosa involving both the deep and superficial mucosa of virtually the entire epithelium. The deep injury induced by these two EtOH concentrations was prevented by both the low and high dose of PGF2 alpha. Of particular importance the 5.0 mg dose of PGF2 alpha provided complete protection (i.e. both superficial and deep) to as much as 50% of the mucosa exposed to 50% or 100% ethanol. These findings indicate that PGF2 alpha possesses "cytoprotective" properties involving both the superficial and deep epithelium that are dose related. PMID- 2980718 TI - GNRH induces activation of Leydig-like cells in Pleurodeles waltlii. A morphometric study. AB - The ultrastructure of the interstitial cells of the glandular tissue of Pleurodeles waltlii was studied in testis of animals obtained in early breeding season (January) under gonadotropic releasing hormone (GNRH) treatments and controls. These cells (parenchymal or Leydig-like cells) displayed the structural characteristics of steroid-producing cells. GNRH administration for 24 hours induced a significant decrease of both medial volume and volume density of lipid droplets. On the other hand, cell volume, nucleus, mitochondria, mitochondrial cristae and tubules of smooth endoplasmic reticulum were increased. The surface density of mitochondrial cristae was also increased. PMID- 2980719 TI - Plasma cell granuloma of the tongue. Report of a case. AB - A case of plasma cell granuloma of the tongue in an otherwise symptomless 48-year old caucasian female is reported. The polyclonal nature of the plasmocytes was revealed by immunostaining of kappa and lambda light chains. Electron microscopic observations showed typical mature plasmocytes. A parasitic etiology of this type of lesion is suggested. PMID- 2980721 TI - Light and electron microscopic study of fetal lung following maternal exposure to methylmercuric chloride. AB - Varying dose levels of methylmercuric chloride (MMC), 1000 ppm (5 mg through 15 mg/kg of body weight), were administered via an intragastric tube to pregnant ICR Swiss/Webster mice on day 9 of gestation. The animals were killed on gestational day 18 and the fetuses removed. Fetal lung sections were processed for light and electron microscopy. A group of animals treated with physiological saline in a similar manner served as the controls. The fetal lungs from treated animals were hypoplastic and retarded in development. The severity of pulmonary changes increased with the dose-levels of MMC. Vacuolation and lysis of mitochondria were seen in fetal lungs. Mitochondrial damage increased in severity with dose-level of methylmercuric chloride. PMID- 2980720 TI - Anti-platelet agents reduce morphological changes of chronic hypoxic pulmonary hypertension. AB - The pathophysiologic mechanism by which chronic hypoxia causes pulmonary hypertension is unknown. If anti-platelet agents, or other pharmacologic interventions, altered the pulmonary vascular changes induced by hypoxia, information concerning the pathogenesis of the pulmonary hypertension or the potential therapeutic usefulness of the drugs might be obtained. In Study 1, rats exposed to chronic hypobaric hypoxia (PB = 520 mmHg) had a pulmonary arterial medial thickness of 6.7 +/- 0.6 mu compared to 4.1 +/- 0.2 mu* for control, normoxic rats (*p less than 0.05). Administration of dipyridamole (2mg/kg/day), or sulfinpyrazone (11 mg/kg/day) in the drinking water reduced the medial thickness to 5.0 +/- 0.3 mu* and 5.4 +/- 0.5 mu* respectively, thus suggesting the possible involvement of platelets in the response of the media to chronic hypoxia. In Study 2, hypoxic rats treated with the calcium blocker, flunarizine, were found to have less medial hypertrophy than a control group of hypoxic rats. This observation suggests that a decrease in transmembrane calcium flux may also reduce medial hypertrophy. PMID- 2980722 TI - The influence of bromocriptine on the ultrastructure of the biceps femoris muscle in mice. AB - Bromocriptine and other dopaminergic agonists drugs are used in Parkinson's disease. In this paper we have studied the ultrastructure of striated muscle of mice after bromocriptine treatment. There was a tremendous increase in the number and size of mitochondria, as well as a very notable increase in the cristae. Some ultrastructure changes were also noted at the neuromuscular junctions. An explanation has been attempted in the light of other investigations concerning the relationship of microtubules and bromocriptine on the one hand, and microtubules and mitochondria on the other. PMID- 2980723 TI - An ultrastructural examination of murine alveolar macrophages following intranasal administration of propionibacterium acnes. AB - Light and electron microscopic analysis of murine lungs or isolated pulmonary cells was performed three days after intranasal administration of the bacterial immunostimulant, Propionibacterium acnes (P. acnes). Our observations indicated that pulmonary alveolar and airway macrophages (PAMs) were the only cells with P. acnes bacilli in their cytoplasm. Bacilli were not observed in pulmonary interstitial macrophages, granulocytes, lymphocytes or pulmonary parenchymal cells such as type I and type II pneumocytes. Because of the morphological heterogeneity of PAMs observed in control and experimental animals, it was not possible from these studies to be certain about the relative abundance or complexity of lysosomes, endoplasmic reticulum, Golgi and other organelles in the two groups. However, we noted that it was not uncommon to observe in the same PAM, profiles of P. acnes and a well developed Golgi complex and endoplasmic reticulum. These P. acnes--associated morphological alterations occurred at a time when functional activities (e.g., phagocytosis, cytostasis) of PAMs were enhanced. PMID- 2980724 TI - Cytostatic action of methylmercuric chloride on mammalian duodenal cells. AB - Adult male mice of the ICR/Swiss Webster strain received a single intragastric administration of methylmercuric chloride 1,000 ppm, at dose levels of 5,10,15,20,25 and 30 mg/kg of body weight. The animals were killed six hours later. Tissue samples from the duodenum were fixed in 10% neutral buffered formalin for light microscopy. Chromosome clumping was observed in dividing cells at all dose levels, resembling a C-mitotic effect. It would lead to reduced mitotic cell formation on account of the subsequent lysis of the arrested metaphases. The cytostatic effect was brought about by the inactivation of the microtubule spindle fiber polymerization mechanism induced by methylmercuric chloride. There was a direct positive correlation between the varying dose levels of methylmercury and the proportion of cells arrested in metaphase in the crypts of the duodenum. PMID- 2980726 TI - Association of sinus histiocytosis with massive lymphadenopathy and idiopathic hypereosinophilic syndrome. AB - A patient was evaluated because of edema, pruritus and generalized painless lymphadenopathy. Laboratory tests showed marked eosinophilia without known etiology. CT scan of abdomen revealed multiple lymph nodes in retroperitoneal area. Lymph node biopsy was reported as sinus histiocytosis, bone marrow biopsy showed hypercellularity with marked infiltration of normal eosinophils. During his admission he developed Coombs positive hemolytic anaemia. Once he was stable, a laparotomy was performed and the patient died two days later because of septic shock. Autopsy revealed sinus histiocytosis with massive lymphadenopathy (SHML) with extranodal involvement of duodenum, spleen and prostate; septic liver and spleen, pyelonephritis, marked infiltration of eosinophils in lymph nodes, spleen, liver duodenum and lungs. To the best of our knowledge, this is the first case report of the association of SHML and Idiopathic Hypereosinophilic Syndrome (HES). PMID- 2980725 TI - Immunohistochemical localization of collagens and fibronectin in human breast neoplasms. AB - Forty four specimens from neoplastic, hyperplastic and normal human breast tissues were studied for localization of collagens and fibronectin. Affinity purified antihuman type I, III and IV collagens and antifibronectins were utilized by the indirect immunoperoxidase technique on fixed and paraffin embedded sections. 86% of the cell cytoplasm of infiltrating ductal and 83% of the lobular cancers were positively stained for collagen type I and III. Collagen type IV, however, was detected in 100% of infiltrating ductal and 83% of lobular carcinomas. Focal cytoplasmic staining is a predominant feature for all antigens in the intraduct carcinoma while a diffuse pattern is encountered in the infiltrating types. Intact basement membranes in various lesions always stained for type IV collagen and showed variable staining for type III collagen and fibronectin. Epithelia of normal, benign, hyperplastic breast and most medullary carcinoma were negative for the three collagen types. Our results are in favour of the view that infiltrating breast carcinoma cells produce inappropriately the majority of collagens and inconsistently other proteins such as fibronectin. PMID- 2980727 TI - Morphological changes in the isolated rat liver perfused in a non-recirculating system: scanning and transmission electron microscopy. AB - Isolated perfused rat livers have been used for various studies, but detailed investigation into the structural integrity of hepatocytes of this system is lacking. In this study, isolated rat livers were perfused in vitro with oxygenated Krebs-Ringer bicarbonate buffer solution, for 2 minutes and 1, 2, 3, and 4 hour(s) at 37 degrees C, using a non-recirculating perfusion system. The perfused livers were processed for semithin section light microscopy, transmission electron microscopy, and scanning electron microscopy. Sectional areas of cell deaths were measured by a camera-tracing assembly from 1.5 microns thick Araldite sections stained with toluidine blue. Progressive nuclear and cytoplasmic changes, leading to cell death, occurred in the hepatocytes of the centrilobular zone, during the 2nd, 3rd, and 4th hour of the perfusion at a rate of 9.03% +/- 1.5%, 38.7% +/- 2.7%, and 55.1% +/- 5.9% (mean +/- standard deviation) of the total sectional areas respectively. Midzonal hepatocytes showed normal basophilic staining but exhibited loss of glycogen granules, loss of microvilli, development of aqueous vacuoles and formation of blebs. The fine structures of cell organelles, glycogen granules, microvilli and plasma membrane of the cells in the periportal zone were well preserved throughout the experimental period. For further quantitative, metabolic and functional studies using isolated rat liver perfused with Krebs-Ringer solution, it is evident from the present investigation that the periportal zone represents the functional region of the hepatic lobule. Whilst progressive changes, leading to cell death, occurred in the centrilobular zone. PMID- 2980728 TI - Glomerular histopathology of the contralateral kidney in experimental unilateral hydronephrosis. AB - The aim of this study was to examine the structural, ultrastructural and morphometric alterations which take place in the contralateral kidneys of rats with experimental unilateral hydronephrosis. 20 Wistar rats weighing 250 g., affected by a process of unilateral hydronephrosis following the ligature of the ureter, were used; these rats were then killed 40, 50, 60, or 70 days after the ligature. Among the perceived alterations, were immunoglobulin G deposits shown by positive immunoperoxidase reaction and increase in the size of the glomerular and corpuscle from around the fortieth day, and structural alterations that included the pedicles, electrondense deposits in the podocytes and pseudogranular structures in the basal membrane of the capillary. PMID- 2980729 TI - Carcinosarcoma of the gallbladder. AB - The case of a 60-year-old woman with a five-year history of abdominal pain, jaundice, and weight loss is presented. On physical examination a hard mass on her right flank was evident. She died under unknown circumstances while she was waiting to be examined. At post-mortem examination the gallbladder was replaced by a neoplasm and there were gallstones within its lumen. Histologically, the tumour was constituted by a mixture of adenocarcinoma, squamous cell carcinoma and undifferentiated sarcoma. The diagnosis of carcinosarcoma of the gallbladder was established. A review of previously reported cases is presented too. PMID- 2980731 TI - Rhabdomyoma of the soft palate. Fine structural details of a highly differentiated muscle tumor. AB - An adult rhabdomyoma was light and electron microscopically studied. The lesional cells presented well-known structural details, such as abundance of mitochondria and glycogen, myofilaments and myofibrils, hypertrophied Z-bands and masses of Z band material. Triads were randomly scattered in the cytoplasm and also related to the individual sarcomeres. In sarcomeres the triads were regularly placed near to the A-I-junctions. This peculiarity of mammalian skeletal muscle fibers may yield a criterion to distinguish between cardiac and extracardiac rhabdomyomas. Circumscribed surface areas of the tumor cells were provided with elaborate infoldings of plasma membrane and basal lamina. These areas were interpreted as imitating myotendinous junctions. Satellite cells were regularly found. PMID- 2980730 TI - Mononuclear cell subpopulations and infiltrating lymphocytes in erythema dyschromicum perstans and vitiligo. AB - Erythema dyschromicum perstans (EDP) and vitiligo are two cutaneous pigmentary dermatoses of unknown etiology. In the present study, the leukocyte infiltrates in the affected skin of EDP and vitiligo patients were studied using the avidin biotin (ABC) immunoperoxidase technique and monoclonal antibodies which recognise the following mononuclear cell subgroups: T-suppressor/cytotoxic (CD8-Leu-2), T helper (CD4 = OKT4), T-suppressor + macrophages (Leu-15), Pan T (CD3 = Leu-4), macrophages (Leu-M3) and Langerhans cells (CD1 = Leu-6), and other cellular markers such as Ia antigens and the Interleukin-2 receptor (CD25 = TAC). The immunocytochemical analysis showed a selective accumulation of CD3+, CD8+, Leu-15 , T-cytotoxic cells in the epidermis of both EDP and early lesions of vitiligo. In addition, an increase in the number of epidermal Langerhans cells (CD1+) was observed in some cases of EDP and vitiligo. The CD4/CD8 ratios in affected and uninvolved skin for both disorders were not significantly different, although values lower than unity were only observed in the infiltrates of affected skin. Ia antigen positivity was observed in the dendritic cells of the dermis and epidermis, as well as in most of the lymphoid cells within the infiltrates for both diseases. Macrophages (Leu-M3) in EDP dermal infiltrates were generally found adjacent to extracellular melanin pigment. Lymphocytes expressing TAC (CD25) surface antigens were also present in the dermal infiltrates. These morphological observations suggest a possible immune cell participation in the dyschromia of such cutaneous disorders. PMID- 2980732 TI - Scanning electron microscopy of human and murine melanoma cells exposed to medium chain-length (C6-C12) dicarboxylic acids in tissue culture. AB - Human and murine (Harding-Passey and Cloudman) melanoma cells were exposed to various concentrations (1 x 10(-3) M-1 x 10(-1) M) of adipic (C6), azelaic (C9), and dodecanedioic (C12) acids for 1-6 hours in tissue culture, and the effects on shape and surface topography were examined by scanning electron microscopy. Effects, i.e., rounding up, concentration of microvilli, blebbing, and prominence of retraction fibrils were time and dose dependent, and for the same concentrations and exposure times, C12 had a greater effect than C9, and both a significantly greater effect than C6. These differential reactions to the three diacids parallel previously reported effects on cell kinetics and viability. The changes could be due to a prime effect on the cell membrane, or they might reflect phases of the cell cycle directed by action of the diacids on the nucleus; this latter seems unlikely. An effect on the cytoskeleton is possibly involved. PMID- 2980733 TI - Primary malignant melanoma of the nasal cavity: report of two cases and review of the literature. AB - Primary malignant melanomas of the nasal cavity are rare, as only 400 cases have been reported to date. The present paper describes two cases recently seen in Caucasian women. The authors point out the difficult clinical diagnosis, as the symptoms are rather aspecific. From the histopathological point of view, diagnosis is easy in the melanotic cases while can show interpretating problems in the amelanotic ones, when melanoma is almost indistinguishable from other malignant neoplasms. A correlation between histological grading and prognosis was not detected, as both cases showed local recurrences within one year after surgery although they were, respectively, of epithelioid and undifferentiated type. While surgery appears to be the choice treatment of the primary lesion, the treatment of cervical metastasis is still disputable. On the whole, most authors think that the role played by radio- and mainly chemo-therapy is still limited and that cervical adenopathies should be treated by a simple lympho-adenectomy rather than by a neck dissection. PMID- 2980734 TI - Ultrastructural study of granulomatous tissue in tonsillar malakoplakia. AB - The ultrastructural study carried out in a case of tonsillar malakoplakia confirmed that granulomatous lesions consisted mainly of macrophages containing peculiar calcified inclusions (Michaelis-Gutmann bodies) considered pathognomonic for the disease. Moreover macrophages frequently contained ingested Gram-negative bacilli and presented aspects of mitochondrial degeneration and autophagy. These latter features were probably the consequence of bacterial infection rather than the expression of primary cellular defects, as the clinical evolution of this case of malakoplakia did not support the existence of generalized macrophage alterations. PMID- 2980735 TI - Smooth muscle distribution in the capsule and trabeculae of the caprine superficial cervical lymph node. AB - This study centers around the dichotomy found in the literature concerning the presence of smooth muscle cells in the trabeculae and capsule of lymph nodes. Various superficial lymph nodes (mammary, mandibular, popliteal, subiliac, and superficial cervical) of the goat were collected and examined by light and electron microscopy. Smooth muscle cells were demonstrated in the capsule and trabeculae of lymph nodes independent of the blood and lymph vessels. PMID- 2980736 TI - Effect of N-ethyl-N-butyl-nitrosamine on the esophageal mucosa of the rat. Histometric investigation of early tumor stages. AB - 80 male Wistar rats received an oral ad libitum application of N-ethyl-N-butyl nitrosamine in a concentration of 0.18 g per litre of drinking water. The changes induced in the esophageal mucosa and determined at three intervals (up to 48 days, up to 91 days, and up to 112 days after commencement of carcinogen exposure) were compared by microscopy with the results from a control group of 10 male Wistar rats of the same age. Several histomorphometric parameters were investigated with the aid of a Leitz ocular micrometer. The earliest localized changes found were an increase in the thickness of the epithelium and the horny layer, and an elongation of the papillary bodies and a widening of the parabasal cellular layer. Later--with a substantial increase in the rate of mitosis in all layers of the epithelium, there was a significant thickening of the non papillomatous and papillomatous epithelium, an enlargement of the nuclei, especially in the middle and upper layers of the epithelium and a thickening of the horny layer, parts of the latter being parakeratotic. The papillomatous changes corresponded in some cases to moderate epithelial dysplasias. As expected, no fully-developed invasive carcinomas occurred in the early period investigated. The histometric data permit the conclusion to be drawn that the lesions described are demonstrable not only at the exophytic-papillomatous epithelium but also in multifocally localized form at the flat, non-papillomatous mucosa, and that they can definitely be regarded as the expression of an incipient field cancerification. PMID- 2980737 TI - Fat-rich diet induces inflammatory changes in the intact rat pancreas. AB - The effects of chronic ethanol ingestion combined with fat-rich, protein-rich or carbohydrate-rich diets on the histology of the intact rat pancreas were studied. 192 male Wistar rats were randomly divided into four different dietary groups. Half of each group received 15% (v/v) ethanol in their drinking solution and the rest were used as controls and given tap water. After a 12-week diet period the pancreas were removed and histological specimens were stained with hematoxylin and eosin. No significant difference was observed between the groups in occurrence of edema, but inflammatory cells were found in (9/24) rats in the fat rich group receiving water (p less than 0.01. In the fat-rich diet group receiving ethanol this finding was observed in 5 of 24 rats. In these groups slight parenchymal cell necrosis was also observed in conjunction with the inflammatory cells. All specimens in the other groups were normal. It is suggested that inflammatory changes caused by a fat-rich diet may be due to unknown toxic effects of this diet. PMID- 2980738 TI - Ultrastructural relationship of quadriceps muscle degeneration with a distant peroneal nerve conduction in human myotonia dystrophica. AB - The association of motor nerve conduction velocity (MNCV) to (1) duration of symptoms, (2) deep tendon reflex responses, (3) clinical muscle atrophy, and (4) ultrastructure of quadriceps muscle was studied in 18 patients with myotonia dystrophica of Steinert and nine normal controls. These patients had neither diabetes mellitus nor any other type of muscle dystrophy. Ultrastructural features of muscle fibers and intercellular spaces between atrophic fibers provided a basis for identifying degenerative changes and evaluating them semi quantitatively. Our study indicates presence of an association between the pattern of muscle degeneration and both MNCV (correlation coefficients, gamma = +0.60) and duration of symptoms (gamma = -0.62), but not between MNCV and duration of symptoms (gamma = +0.28). Further analysis of the association between the degeneration of quadriceps and the MNCV of a distant peroneal nerve (which does not innervate quadriceps) suggested that the systemic nerve degeneration occurred in some groups of myotonia patients. Our study indicates that while in some patients the muscle degeneration may have been associated with the impairment of neurogenic elements, in others it occurred in the absence of any MNCV abnormality. Our findings favor the role of both neuropathic and myopathic factors in the muscle degeneration seen in myotonia dystrophica. PMID- 2980739 TI - A study of myonecrosis induced by the venom of the scorpion tityus serrulatus. AB - The pathogenesis of skeletal muscle necrosis produced by Tityus Serrulatus venom was studied by means of light microscopy and electron microscopy. Wistar rats were inoculated subcutaneously, at some distance from the muscles under study, with a sublethal dose of scorpion venom. Samples were taken of the tibialis anterior muscles of both rear legs, 2, 7 and 24 hours postinoculation. Light microscopy analysis after 2 hours revealed certain changes identified as "delta lesions", and also the presence of hyperconcentrated muscle cells. Electron microscopy confirmed these lesions and also enabled us to identify a degree of discontinuity in the plasma membrane with a persistence of the basal membrane. Hyperconcentrated fibers could still be observed 7 hours postinoculation. Histochemical analysis revealed high levels of calcium within the fibers. 24 hours after inoculation with the venom, numerous phagocytic cells were found in the degenerated fibers. Muscle cells were also found to have undergone alterations indicative of an ischemic process. The most characteristic finding 7 days postinoculation was the appearance of regenerative fibers. After thirty days the muscles regained their normal appearance. It is suggested that Tityus Serrulatus venom induces myonecrosis by means of a twofold action: direct action, which gives rise in the first place to a rupture of the plasma membrane, permitting a massive entry of calcium this being a key factor in the process of cell lesion and an assumed indirect action due to ischemia. PMID- 2980740 TI - Thermotherapy for Harding-Passey melanoma: light and electron microscopic study. AB - The changes of implanted Harding-Passey melanoma in C57Bl/6J mice following treatment with wholebody hyperthermia were studied. The treated tumours showed a progressive growth delay, cellular and architectural irregularities as well as cell injury characteristics. The presence of distended and irregular blood vessels, the peripheral localization of the melanosomes and the melanosome complexes were constant. PMID- 2980741 TI - Electron microscopic studies on the pars intermedia of the rat under stress. AB - Using electron microscopy, we have studied the pars intermedia (PI) of the pituitary gland of Wistar rat under psychogenic stress, finding a considerable increase in dark cells. The cells of the PI present striking vacuoles in all their cytoplasms, increasing up to a phase of total cellular disintegration. We have also observed a cell type, not found in the animal control group, of regular size and electrondense matrix of medium contrast. PMID- 2980742 TI - Ultrastructure of the tubular nephron of Testudo graeca (Chelonia). A comparison between hibernating and non-hibernating animals. AB - The tubular nephron of hibernating and non-hibernating specimens of Testudo graeca (Chelonia) was studied by means of conventional light and electron microscopy and histochemistry. The tubular nephron was composed of proximal, intermediate, distal and collecting tubules in both hibernating and non hibernating animals. The cells of the proximal tubule showed long microvilli, cytoplasmic vacuoles, a developed endoplasmic reticulum and abundant mitochondria. Fat droplets were also observed. The intermediate segment was lined by ciliated and non-ciliated cells. The lining cells of the distal tubule presented few microvilli, abundant dense mitochondria and clear vesicles of mucous appearance in the terminal portion. Collecting ducts are composed of mucous and non-mucous cells. Mucous cells presented strong reaction to the histochemical techniques detecting sialo- and sulpho-mucins. During hibernation, a progressive vacuolar degeneration of the endoplasmic reticulum was observed in all the segments of tubular nephron, which may be caused by a massive intake of extracellular water into the cell. PMID- 2980743 TI - Interstitial collagen in alcoholic human liver. AB - The occurrence and intensity of staining for specific antibodies against the aminoterminal propeptide of type III procollagen (PIIIP), which is indicative of the synthesis and the degradation of that collagen type, was studied in sections from normal and alcoholic livers and compared with serum PIIIP levels, serum antipyrine clearance, fibronectin distribution and morphology as revealed by conventional stains and electronmicroscopy. Positive staining for PIIIP and fibronectin was observed in the perisinusoidal space of the normal liver and in portal tracts. In alcohol-induced fatty liver positive staining increased around the central veins, in alcoholic hepatitis increased staining reaction was seen to a limited extent in areas of cell injury. Extensive reticulin and PIIIP-positive areas were found in the periportal interstitium of the cirrhotic livers and in large fibrotic areas extending into the surrounding parenchyma in cases of active disease. The results show a distinct relationship between collagen type III metabolism, morphologically detectable hepatic injury and liver cell function tests, with tissue deposition occurring later in the disease process than biochemically detectable serum collagen levels and signs of altered liver cell function. PMID- 2980744 TI - Secondary amyloidosis and cystic fibrosis. A morphological and histochemical study of five cases. AB - The pathological findings of five cases of amyloidosis associated with Cystic Fibrosis are reported. Rectal biopsy led to the diagnosis of amyloidosis in four patients. In three cases the diagnosis was confirmed at autopsy, with involvement of spleen, liver, kidneys, adrenal glands, thyroid and other organs. It seems that Secondary Amyloidosis provokes a significant, although rare, complication of Cystic Fibrosis as greater numbers of these patients survive into adulthood. PMID- 2980745 TI - Immunohistochemical change of actin in experimental myocardial ischemia. Its usefulness to detect very early myocardial damages. AB - Pathomorphological diagnosis of acute myocardial infarction has many problems in human autopsy materials less than 4 to 6 hours after clinical onset and in rats 2 to 3 hours after experimental coronary occlusion. Since immunohistochemical reaction depends on the antigen determinant site of the material, changes in the reaction may reflect alterations at the molecular level in myocardial fibers. With this consideration in mind, the effectiveness of diagnosing infarction at the earliest (possible) stage, and the changes of actin filaments were investigated through experiments, using immunohistochemical methods involving anti-actin antibodies produced from chicken gizzards in our laboratory. The left coronary arteries of rats were ligated to produce ischemia. Dehydrogenases were shown to be still present by triphenyltetrazolium chloride (TTC) reaction, but the anti-actin antibody reaction had disappeared in areas corresponding to the ischemic sites. However, on electron microscopic examination of these sites, actin fibers were clearly revealed. In the case of ischemia lasting for more than 6 hours, the anti-actin antibody reaction had disappeared, corresponding to the disappearance of the TTC reaction. At this stage, myocardial actin fibers were revealed by electron microscopic examination. These results indicate that ischemia induces some type of biochemical degeneration at the molecular level of myocardial actin, most likely the change of actin polymerization. Moreover, they show that the anti-actin antibody technique is capable of detecting such very early degenerative and ischemic changes proving itself to be better suited for determining the range and degree of early infarction. PMID- 2980746 TI - Morphometric study of hepatic ultrastructure in alcoholic hepatitis. Veterans Administration Cooperative Study Group on Alcoholic Hepatitis. AB - We undertook a morphometric analysis of hepatocellular organelles in an attempt to correlate their changes with the clinical stages of patients with alcoholic hepatitis. Although hepatic ultrastructural alterations did not correlate with disease severity, we found significant differences between patient and control groups in the measured parameters of non-organelle cytoplasm, mitochondria, SER, RER, glycogen, and lipid. PMID- 2980747 TI - Photoreceptor fine structure in the vervet monkey (Cercopithecus aethiops). AB - The structure of the photoreceptors of the vervet monkey (Cercopithecus aethiops) has been investigated by light and electron microscopy. In this species the photoreceptors can be readily differentiated and adequately described by the classical terminology of rods and cones, with rods being the more numerous. Rods are long, slender cells while cones are shorter and stouter. Both rods and cones are highly differentiated cells and consist of an outer segment, a connecting cilium, and inner segment, a nuclear region and a synaptic process leading to a synaptic ending. Morphological differences are noted between rods and cones for the various regions of these cells. PMID- 2980748 TI - Alcohol cardiomyopathies. PMID- 2980749 TI - Percutaneous endoscopic gastrostomy: new management of a difficult problem. AB - Under appropriate conditions, enteral feeding is the preferred method of nutritional management. Nasoenteric tube feeding may be adequate for short-term use but a more permanent indwelling gastrostomy tube is favored when long-term nutritional support is planned. PEG has several advantages over the surgical approach and should be considered the procedure of choice. PMID- 2980750 TI - Experience with long-term indwelling central venous catheters in children. PMID- 2980751 TI - How your practice can cope with change. PMID- 2980752 TI - Challenges to private practice: the Deficit Reduction and the Civil Rights Acts of 1984. One person's opinion. PMID- 2980753 TI - Survival in the paper jungle, or how to read medical journals. PMID- 2980754 TI - University hospital in transition. PMID- 2980755 TI - Hepatitis B virus-associated membranous glomerulonephropathy. AB - A variety of extrahepatic abnormalities have been described in patients with hepatitis B viral infection, including the prodromal arthritis (serum sickness like) syndrome, polyarteritis nodosa, and glomerulonephropathy. Five patients with hepatitis B virus (HBV)-associated membranous glomerulonephropathy (MGN) were persistent carriers of HBsAg. The deposition of HBsAg in the renal lesions of these patients was proved by using monospecific antibody and immunofluorescence technique. These findings support the hypothesis of an etiological correlation of hepatitis B viral infection with some cases of MGN. HBV-associated MGN may be much more frequent in the world than previously reported, and more cases must be collected for the understanding of this unique entity. PMID- 2980757 TI - In the eye of the needle. A reappraisal of endoscopic sclerotherapy. AB - Endoscopic sclerotherapy is routinely used as treatment for bleeding esophageal varices. Recent controlled trials have evaluated the impact of sclerosis on short and long term survival after varices hemorrhage and compared sclerotherapy with shunt surgery. These studies are reviewed and the future of sclerotherapy discussed. PMID- 2980756 TI - Problems with clinical trials on Campylobacter pylori. AB - Clinical trials investigating Campylobacter pylori infection have proven difficult to conduct and to interpret. Those trials examining the relapse rates of duodenal ulcer disease among subjects in whom C. pylori infection has been eradicated have shown a significant reduction in recurrence rates. However, since the Bismuthate compounds used in these trials have cytoprotective ulcer-healing properties, reduced relapse rates cannot be attributed to antibiotic activity alone. Those trials examining infective gastritis and "nonulcer dyspepsia" are hampered by the ill-defined nature of this entity, "dyspepsia," and by the subjective end-points of these trials. PMID- 2980758 TI - The future of invasive pancreaticobiliary endoscopy. AB - The futures of invasive pancreaticobiliary endoscopy will be contigent not only on what can be technically accomplished through the endoscope, but upon competing or complementing technologies. Thus, widespread application of extracorporeal shock wave lithotripsy (ESWL) for gallstones has the potential for either dramatically increasing or decreasing the endoscopic workload. Use of miniscopes, biliary stone extraction without sphincterotomy, and endoscopic approaches to benign pancreatic disease are all possible future scenarios. PMID- 2980759 TI - Current peptic ulcer time trends. An epidemiological profile. AB - There has been speculation that peptic ulcer is a vanishing disease. To determine current time trends for peptic ulcers, we examined data from the National Center for Health Statistics and the Commission on Professional and Hospital Activities covering the period 1970 to 1985. For duodenal ulcer, hospitalizations for uncomplicated cases continue to decline, but there has been no decrease in cases with complications (such as perforations or hemorrhages). Hospitalizations for gastric ulcer are increasing, especially for cases with hemorrhage. Duodenal and gastric ulcer death rates have remained fairly constant since 1979. Site unspecified peptic ulcer death rates doubled in 1979 and have remained at this rate through 1983. This increase can largely be attributed to coding changes made in the 9th revision of the International Classification of Diseases. PMID- 2980760 TI - Effects of microencapsulated vs. enteric-coated acetylsalicylic acid on gastric and duodenal mucosa: an endoscopic study. AB - Two preparations of acetylsalicylic acid (ASA) were tested for their effects on gastroduodenal mucosa in a randomized crossover double-blind study that involved 12 healthy volunteers. Medication M (Monobeltin) consisted of 1,050 mg ASA provided with an enteric coat and medication C (Colfarit) consisted of 1,000 mg of a microencapsulated ASA preparation. Both drugs were taken for 6 consecutive days each. Upper gastrointestinal endoscopy was performed before and 2 and 6 days after each medication. An interval of 10 days elapsed between one treatment and the other. Gastric lesions occurred in 10 of 12 subjects taking medication M and in 11 of 12 subjects taking medication C after 2 days and were present in all subject after 6 days. Duodenal lesions were seen only in subjects taking medication C. The degree of gastric mucosal lesions based on two grading scales was not significantly different between the two treatments. Neither of the two pharmaceutical formulations of ASA provided sufficient protection for the gastric mucosa. PMID- 2980761 TI - Patients with pulse rate changes in irritable bowel syndrome. Further evidence of altered autonomic function. AB - Patients with the irritable bowel syndrome have significantly more rapid and significantly greater slowing of the pulse rate than do age-, sex-, and stressor matched controls when their pulse rate is measured at initial outpatient clinic attendance. These findings provide further evidence that altered autonomic function exists in patients with the irritable bowel syndrome. PMID- 2980762 TI - Narcotic dependence in inflammatory bowel disease. AB - Thirteen of 43 patients (30%) with inflammatory bowel disease referred for psychiatric consultation were found to be drug dependent, most commonly on oral narcotics. Drug dependence was more frequent in patients with Crohn's disease than ulcerative colitis and many had a borderline personality disorder. The study suggests that drug dependence is not recognized often enough in patients with inflammatory bowel disease and that patients with certain psychiatric disorders are at higher risk of developing it. Recognition of drug dependence is aided by interviewing family members. It is best prevented by seeking and treating the specific cause of pain and by having only one physician assigned to prescribe and manage narcotics. PMID- 2980763 TI - Crohn's disease and Dalziel's syndrome. A history. PMID- 2980764 TI - Historical aspects of inflammatory bowel disease. AB - Ulcerative colitis and regional enteritis (Crohn's disease), probably first appeared as isolated cases, several centuries ago; but not until the latter half of the 19th century did their clinical presentation attract medical interest as definable entities. This review of the early medical literature reveals a surprising number of reports of inflammatory bowel disease consistent with present descriptions of these conditions. As with other diseases, the steadily increasing numbers of patients with ulcerative colitis during the first third of the 20th century and of patients with regional enteritis during the latter two thirds of the century follows a familiar pattern encountered with diseases wherein environmental factors play a pathogenetic role. The early history of ulcerative colitis is lost in the complex mixture of infectious colitides, only partially unraveled in the past 100 years. The early history of regional enteritis probably is linked with the numerous European and American reports of tumor-like granulomas of the small intestine, initially regarded as neoplastic lesions requiring surgical removal. It now seems possible that physicians over a period of several centuries may have encountered these conditions but were not in a position to appreciate their uniqueness. The careful accounts of many authors, especially Morgagni (1761), Wilks (1859), Fenwick (1889), Dalziel (1913) and of Crohn, Ginzburg, and Oppenheimer (1932) of then obscure intestinal inflammatory processes thus re-emphasizes the importance of meticulous clinical and pathological descriptions in the subsequent elucidation of obscure disorders. If the course of ulcerative colitis and of Crohn's disease follow the historical pattern of other once important and later disappearing human illnesses, we may have reason to anticipate clarification of their etiology and possible relationship in the not too distant future. PMID- 2980765 TI - The rise and fall of the psychosomatic hypothesis in ulcerative colitis. AB - We review the rise, in the 1930s, and the decline, in the 1960s, of medicine's attention to psychosomatic considerations in ulcerative colitis. Both the rise and decline of interest in psychosomatics resulted from trends outside of the published work which directly contested the issue. In particular, the "rise" period coincided with the heyday of the psychosomatic movement and the "fall" with the autoimmune theory, the introduction of steroids, and other trends. Finally, we offer a general critique of the way the debate over the psychosomatic hypothesis was conducted. PMID- 2980766 TI - Quality of life in patients with inflammatory bowel disease. AB - To investigate the effect of inflammatory bowel disease (IBD) on the quality of life, we interviewed 43 patients with ulcerative colitis (UC) and 54 with Crohn's disease. Patients identified frequent and important problems in five areas. Primary bowel symptoms, systemic symptoms, and altered emotional function were common; functional and social impairment were less frequent. Systemic symptoms such as fatigue were more prevalent in patients with Crohn's disease. Apart from primary bowel complaints, patients seldom volunteered other facets of quality of life impairment; this was particularly true for impairment of emotional function. We conclude that despite troublesome intestinal and systemic symptoms, most patients with IBD avoid major disruption in work and personal lives. Physicians must inquire specifically about emotional problems relating to IBD; in particular, fear of surgery is important to address. Psychosocial interventions should be targeted to those patients with problems in this area. PMID- 2980768 TI - Choledochal cysts in the adult. AB - Although choledochal cyst is generally considered a disorder of childhood, in approximately 20% of patients a cyst is first recognized in adult life. We reviewed nine adults who underwent operation for choledochal cyst from 1974-1985 at Yale New Haven Hospital. Presenting symptoms included: crampy right upper quadrant pain, jaundice, pruritus, cholangitis, pancreatitis, hepatomegaly, and elevated liver function tests. Seven patients had undergone previous biliary surgery. The eight patients with type I cysts underwent complete excision of the cysts with Roux-Y choledochojejunostomy. One patient with a type IVa cyst underwent Roux-Y cystjejunostomy. All patients had complete resolution of biliary symptoms. There were no deaths, serious complications, or development of biliary malignancy in the 6 months to 13 years of follow-up. Optimum treatment of choledochal cyst is complete excision with reconstruction by Roux-Y choledochojejunostomy. PMID- 2980767 TI - Cancellation rates and gas scores for air contrast barium enema immediately after 65-CM flexible sigmoidoscopy. A randomized clinical trial. AB - A randomized prospective study examined the impact of bowel gas introduced by 65 cm flexible sigmoidoscopy (FS) on the ability to perform air-contrast barium enema (ACBE) on the same day. Seventy-five patients at risk for colorectal cancer were randomly assigned to two groups. Of these, 28 patients in each group completed the protocol. The study group received flexible sigmoidoscopy and air contrast barium enema on the same day, whereas the control group had their air contrast barium enema on a different day. Bowel gas observed on an abdominal scout film prior to air-contrast barium enema was quantified on a scale of 1 (excessive gas) to 5 (no gas). The cancellation rate for the air-contrast barium enema was measured in each of the groups. The study group had significantly more bowel gas compared with the control group (p = 0.000003). The air-contrast barium enema cancellation rate was also higher in the study group (36%) than in the control group (14%) (p = 0.06). Greater than 60% of the study group patients were successfully examined. Same-day scheduling would reduce the number of bowel preparations required in the evaluation of patients at risk for colorectal cancer. Although retrospective studies have suggested no impact of same-day FS on ACBE quality, radiologists in this study canceled approximately one-third of scheduled patients due to perceived excessive bowel gas. PMID- 2980769 TI - Pemphigus vulgaris of the esophagus. AB - We describe a case of pemphigus vulgaris involving the lower esophagus, in a patient with gastroesophageal reflux. As near as we can tell, this is the first reported case of esophageal involvement by pemphigus in the absence of oropharyngeal disease. Gastroesophageal reflux may have precipitated the development of esophageal pemphigus. PMID- 2980770 TI - Isolated gastric sarcoidosis. Unique remnant of disseminated disease. AB - Isolated granulomatous disease was identified in a gastrectomy specimen after a gastrointestinal hemorrhage in a 61-year-old white woman. She died postoperatively. No other areas of granulomatous tissue were identified at autopsy. Ten years previously, pulmonary sarcoidosis was proven on lung biopsy. Conceivable, other reported cases of unexplained "idiopathic" granulomatous disease of the stomach may represent residua of prior disseminated sarcoidosis. PMID- 2980771 TI - Squamous morules in gastric mucosa. AB - An elderly white man undergoing evaluation for pyrosis was found to have multiple polyps in the fundus and body of the stomach by endoscopic examination. Histologic examination of the tissue removed for biopsy over a 2-year period showed fundic gland hyperplasia and hyperplastic polyps, the latter containing nests of immature squamous cells. These nests arose within superficial gastric glands and resembled squamous morules seen in benign endometrium. Previous reports of this type of squamous epithelium in gastric mucosa were not encountered in our literature review. This most probably represents a benign process since there is no evidence to support its relationship to the occurrence of either primary squamous cell carcinoma or adenosquamous carcinoma of the stomach. PMID- 2980772 TI - Carcinoma of the gallbladder from an anomalous connection between the bile duct and the pancreatic duct. AB - Anomalous connection between the common bile duct and the pancreatic duct has recently been reported to be associated with cancerous changes of the biliary tract. A 52-year-old Chinese man had a fungating tumor in the gallbladder, and a long common channel between the bile duct and the pancreatic duct. The association of these two rare disease entities especially in a Chinese man may imply a close relationship between pancreatobiliary anomalous connection and the gallbladder cancer. It is thus prudent to monitor patients with such an anomalous connection closely by serial ultrasonic examination. More aggressively, prophylactic cholecystectomy and reconstruction of the biliary tract may be indicated. PMID- 2980773 TI - Recurrence of hepatic amebiasis after successful treatment with metronidazole. AB - A patient with an amebic liver abscess had an incomplete response to an initial course of metronidazole, but subsequently responded to a prolonged course of the agent. Recurrence of disease in the contralateral hepatic lobe responded to chloroquine along with a luminicide. We took the initial failure of metronidazole to be secondary to either the advanced nature of the disease or the development of partial resistance to metronidazole. The recurrence was the result of reinfection of the liver by persistent organisms in the bowel, emphasizing the importance of giving a lumicide in the initial treatment of this lesion. PMID- 2980774 TI - Focal segmental ischemia of the terminal ileum mimicking Crohn's disease. AB - The clinical, radiographic, and pathologic features of focal segmental ischemia are similar to Crohn's disease. We report a patient with focal segmental ischemia mimicking Crohn's disease and discuss the histologic distinction between the two entities. Ischemia must be considered when "Crohn's-like" lesions are encountered in elderly patients. PMID- 2980775 TI - Resection of hepatic metastases in a patient with Crohn's carcinoma. PMID- 2980776 TI - Crohn's disease of the esophagus: further considerations. PMID- 2980777 TI - Acute cholecystitis complicating ERCP: a case report. PMID- 2980778 TI - Nonspecific ulcer of the jejunum. PMID- 2980779 TI - Protein design using model synthetic peptides. AB - We have designed and synthesized a small, unique protein molecule with defined secondary, tertiary and quaternary structure. This 35-residue peptide, containing a cysteine residue at its N-terminal end, was oxidized to form a 70-residue disulfide-linked two-stranded alpha-helical coiled-coil with the two alpha helices parallel and in-register. The major contribution to the formation and stabilization of the alpha-helical coiled-coil is hydrophobic interactions between positions 2 and 5 of the heptapeptide repeat (Lys-Leu-Glu-Ala-Leu-Glu Gly). The protein (L-protein) contains nine leucine-leucine hydrophobic interactions between the alpha-helices of the coiled-coil. Circular dichroism studies demonstrated that this protein in its reduced ([L (r)] or oxidized (L (o)] state was essentially 100% alpha-helical ([theta]220 = -34,050 and -32,000 degrees respectively) at pH 2 (0.1% aqueous trifluoroacetic acid). Our objective was to modify systematically the structure of L to delineate the contribution that various amino acid side chains make to the formation and stabilization of its three-dimensional structure. A-protein, which contains alanine instead of leucine at positions 16 and 19 of the hydrophobic repeat in each chain of the coiled-coil, was compared to the L-protein. At pH 2, the oxidized form of the A protein [A (o)] was essentially 100% helical. However, the protein was much less stable to temperature denaturation compared to the L-protein. The replacement of two leucine-leucine interactions by two alanine-alanine interactions has a dramatic effect on the formation and stability of the two-stranded alpha-helical coiled-coil structure. The results of this study clearly demonstrate the validity of this synthetic model protein approach to understanding the molecular aspects responsible for the folding and stabilization of protein molecules. PMID- 2980780 TI - An overview of desorption ionization mass spectrometry in peptide and protein analysis. PMID- 2980781 TI - Mechanisms of aspartimide formation: the effects of protecting groups, acid, base, temperature and time. AB - Factors affecting aspartimide formation, such as protecting groups, acidity, basicity, and temperature, were studied using the model tetrapeptide, Glu-Asp-Gly Thr. The aspartyl carboxyl side chain in this tetrapeptide was either free or protected as a benzyl or cyclohexyl ester. Our results showed that the cyclohexyl ester led to far less aspartimide formation during acidic or tertiary amine treatment than the corresponding benzyl ester. The rate constants of aspartimide formation in HF-anisole (9:1, v/v) for the tetrapeptide protected as the benzyl ester were found to be 6.2 x 10(-6) and 73.6 x 10(-6) s-1 at -15 degrees and 0 degrees C respectively. These values were about three times faster than the corresponding free- or cyclohexyl ester-protected tetrapeptide. Little difference was seen when the studies were carried out at room temperature. The cyclohexyl protected tetrapeptide gave only 0.3% aspartimide in diisopropylethylamine treatment in 24 h, a 170-fold reduction of imide formation when compared with the benzyl protected tetrapeptide. Thus, using the cyclohexyl ester for aspartyl protection, our studies showed aspartimide formation could be significantly reduced to less than 2% under standard peptide synthesis conditions. Furthermore, with these model peptides, the mechanism of acid catalyzed aspartimide was studied in a range of HF concentrations. In dilute HF cleavage conditions (HF:dimethylsulfide 1:3, v/v), the mechanism was found to be of the AAC2 type, with the rate of aspartimide formation increasing very slowly with increasing acid concentration. In concentrated HF solutions (HF greater than 70% by volume), the rate of aspartimide formation increased rapidly with the increase in acid concentration. However, from model studies, the mechanism of aspartimide formation in concentrated HF was AAC2 rather than AAC1. PMID- 2980782 TI - A new tool for peptide separation. AB - A family of reagents has been developed (Pep-Seps) which provide separations of peptides based upon the presence or absence of a single type of amino acid. This separation is achieved through a reversible, covalent attachment of the peptide to the Pep-Seps reagents. Pep-Seps provide a more specific separation than most forms of HPLC and may be used to complement HPLC in peptide separations and isolations. Several specific applications will be discussed. PMID- 2980783 TI - The Magainins: sequence factors relevant to increased antimicrobial activity and decreased hemolytic activity. AB - The Magainins, two antimicrobial peptides found in the skin of the frog Xenopus laevis, and 50 Magainin analogs were synthesized by the method of simultaneous multiple peptide synthesis (SMPS). This series of peptides was prepared in order to examine the effects of omitting individual amino acids on antimicrobial activity. The series consisted of 22 Magainin 1 omission analogs having a C terminal carboxyl (M1-C) and 23 Magainin 2 omission analogs having a C-terminal amide (M2-A), as well as both the C-terminal amide and carboxyl forms of Magainin 1 and Magainin 2. These peptides were tested against E. coli (Gram negative), S. epidermis (Gram positive) and C. albicans (yeast). Amino acid omissions in the N terminal region (residues 1-14) resulted in the complete loss of antimicrobial activity in both Magainin series. These analogs also had very low hemolytic activity against human erythrocytes. However, analogs with omissions in the C terminal region, especially residues alanine-15, glycine-18 or glutamic acid-19, while having equal or increased antimicrobial activity relative to the original Magainin 1 or Magainin 2 forms, had variable hemolytic action. Thus, both Magainin 1 and Magainin 2 with the glutamic acid 19 omission had equal activity against E. coli and increased activity against S. epidermis, while having lower hemolytic activity than the original sequences. The amide form of Magainin 2 with glycine 18 omitted had equal antimicrobial activity, but significantly increased hemolytic activity. The C-terminal carboxyl form of Magainin 1, however, showed equal antimicrobial activity, but substantially decreased hemolytic action.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980784 TI - Isotope-edited NMR studies of Fab'-peptide complexes. AB - Complexes formed between a monoclonal anti-peptide Fab' and its complementary 15N labeled peptide were studied primarily by isotope-edited nuclear magnetic resonance (NMR) techniques. The monoclonal antibodies used were raised against peptides corresponding to residues 69-87 of the protein myohemerythrin. The complexes studied correspond to Fab' bound to the synthetic peptide, MHKDFLEKIGGL NH2 (residues 76-87 of myohemerythrin) labeled with 15N at various amides. The combined approach of using specifically 15N-labeled peptides and reverse detection solution NMR techniques has allowed us to monitor selectively the peptide component of the Fab'-peptide complexes studied. Through the use of these techniques, we have been able to directly observe the resonances of the bound peptide, as well as distinguish between free and bound peptide resonances in situations when excess peptide is present. NMR titrations of the Fab' with antigen have shown that there is one site to which the peptide strongly binds. The NMR parameters of the various residues examined thus far have been quite distinctive from each other, reflecting the ability of these techniques to detect differences in the local environment of the individual residues. Differences with respect to these parameters among the various residues may be related to the overall antigenic properties of the peptide. PMID- 2980786 TI - Effects of trifluoperazine and membrane-bound sialic acid on 45Ca2+ uptake into erythrocytes. AB - After removal of more than 70% of the sialic acid from the erythrocyte membrane by treatment with neuraminidase, trifluoperazine (TFP) increased the rapid and slow phase of 45Ca2+ uptake into erythrocytes, identically to Ca2+ uptake into untreated erythrocytes from patients with tetanic syndrome. Both phases of 45Ca2+ influx were inhibited by extracellular Mg2+, Ki amounted to 4 mmol/l. However, the increased 45Ca2+ influx into patient erythrocytes in the presence of TFP was not caused by a reduced sialic acid content in the patients' erythrocytes. It was concluded that erythrocytes from patients with tetanic syndrome either bind more TFP or, if the same amount of TFP is bound, it has a stronger effect on 45Ca2+ influx. PMID- 2980785 TI - Ion-exchange HPLC for peptide purification. AB - Ion exchange HPLC is a very useful but underutilized tool for peptide purification. Ion exchangers provide separations that are complementary to reversed-phase HPLC. Purification procedures utilizing both ion exchange and reversed-phase modes have greatly increased resolving power compared to procedures that use reversed-phase alone. This increased resolution is particularly useful for the purification of natural peptides from complex tissue extracts and for synthetic peptides from the many structurally similar byproducts of synthesis and HF cleavage. The high capacity of ion-exchange HPLC columns allows them to be used at the front end of a purification scheme, reducing the sample load on subsequent reversed-phase columns. In addition to improving overall resolution in a purification scheme, the charge selectivity of ion exchangers solves specific resolution problems involving charge differences between peptides. PMID- 2980787 TI - Concentration of selected metals in normal and pathological human seminal plasma. AB - The levels of zinc, ultrafilterable (UF) zinc, copper and manganese in normal and pathological (astheno-, azoo-, oligospermic) seminal plasma were measured by flameless atomic absorption. For ultrafiltrable zinc, a special method was used to treat Amikon membranes. A lower ultrafiltrable zinc level was found in azoospermic and a lower manganese level in oligospermic seminal plasma. At this time the meaning of these results is unknown. PMID- 2980788 TI - Retention of intratracheally instilled and ingested tetravalent and pentavalent vanadium in the rat. AB - The retention, subcellular distribution in the liver, kidneys and lungs and binding of vanadium in the rat were investigated at 3 h, 1 d and 12 d after intratracheal instillation of 200 ng/kg body weight of 48V-labelled pentavalent and tetravalent vanadium. The metabolic patterns of both chemical forms of vanadium were similar. The lungs, liver, kidneys, bone, testes and spleen are the target tissues at risk, vanadium being removed from them with time at different rates. Pulmonary vanadium clearance was initially rapid - 80-85% of the 48V was removed within 3 h. At 12 d about 2% of the element was still present in the lung. Intracellularly, the major part of liver, kidney and lung vanadium was present in the nuclear fraction (30-40% of the homogenate), followed by cytosol and mitochondrial fractions. Gel filtration chromatography of the lung cytosol showed two biochemical pools of vanadium: the first corresponding to protein bound vanadium, which may be involved in the long-term accumulation of the element in the lung, the second pool representing a diffusible vanadium form. The retention of the tetravalent and pentavalent vanadium forms was also investigated 1 d after oral administration. No obvious differences were observed in the distribution pattern of 48V in the tissues, the retention factor being higher by two orders of magnitude in comparison with the intratracheally instilled animals. These findings suggest that the metabolic pathways of tetravalent and pentavalent vanadium are independent of the route of vanadium exposure. PMID- 2980789 TI - Aluminum determination in whole blood, dialysis solution, and tap water samples from Maracaibo dialysis units (Venezuela) by graphite furnace atomic absorption spectrometry. AB - Patients with chronic renal failure (CRF) on periodical hemodialysis may accumulate aluminum in tissues and show typical disorders such as dialysis encephalopathy, osteodystrophy, and microcytic anemia. Aluminum contamination of the water used to prepare the dialysis solution (dialysate) is one of the metal sources that may affect people under hemodialysis, especially in units in which untreated water is used. Graphite furnace atomic absorption spectrometric methods for aluminum determination in whole blood, dialysis solution, and tap water samples from CRF patients were developed, based upon the use of the same furnace temperature program. Samples were diluted 4-fold with 0.6% triton X-100 (whole blood) or with 0.01 mol/L nitric acid (dialysis solution and tap water) and analyzed by aqueous standard (blood and tap water) or matrix-matching standard (dialysis solution) calibration curves. The characteristic masses were 33.8, 11.3, and 19.5 pg Al/0.0044 A.s for whole blood, dialysate, and tap water, respectively. In the diluted solutions, the detection limits (2 sigma) for the described methods were 0.5 microgram/L Al (whole blood), 0.4 microgram/L Al (dialysate), and 0.4 microgram/L Al (tap water). The methods were applied to samples from several CRF patients under hemodialysis at Maracaibo University hospital. The data revealed extremely high aluminum levels, which corresponded to the symptoms of dialysis encephalopathy and/or osteodystrophy showed by some of them. The proposed methods are reliable and reproducible. PMID- 2980790 TI - Concentration of nickel and chromium in serum: influence of blood sampling technique. AB - In the determination of nickel and chromium in serum, contamination is the most important source of error. In this study the influence of needle type and collection device was investigated. For each of 20 subjects the concentrations of nickel and chromium in serum obtained by the following techniques of blood collection were compared: closed collection system (acidwashed polyethylene syringes) + (A) steel needle (Terumo), (B) teflon i.v. cannula (Abbocath-T), and (C1) teflon i.v. cannula (Venflon); (C2) open collection system (acidwashed polyethylene tubes) + teflon i.v. cannula (Venflon). For each technique the first 5 ml of blood were discarded. No statistically significant differences were found between serum concentrations of nickel and chromium in specimens collected with the sampling techniques A, B, C1 and C2. This study has shown that identical results for concentrations of nickel and chromium in serum are obtained independent of whether a steel needle or a teflon i.v. cannula is used for blood sampling, provided the first 5 ml of blood are discarded. Likewise, identical results are obtained independent of whether specimens are collected in a closed or an open collection system, provided collection devices are acidwashed and sampling takes place under hospital conditions. PMID- 2980791 TI - Avoidance of contamination in element analysis of serum samples. AB - The selection of sampling needles and plastic material used for the storage of serum samples were studied. Sampling needles and plastic material were analyzed by neutron activation analysis. Leaching of elements from containers by acids and EDTA-solution was also followed. Also the long-term storage of serum samples for element analysis was investigated. Zn, Cu and Se concentration of serum was followed over the course of four years. The selection of sampling needles, as well as all plasticware which comes into contact with the serum, should be based on the knowledge of possible removal of elements from materials into the sample. Storage of serum samples at -18 degrees C does not change the copper, zinc or selenium concentration over the course of 4 years despite repeated melting and refreezing of the samples. PMID- 2980792 TI - Metallothionein gene expression in mouse tissues by D-penicillamine. AB - The effect of D-penicillamine on metallothionein mRNA accumulation was examined in mouse tissues by Northern and dot blot analysis. This drug was given as a single intraperitoneal dose of 250 mg/kg body weight and the metallothionein mRNA content of the tissues was measured 1, 4, 8 and 24 hours later. A detectable increase of mRNA was observed after 1 hour and maximal accumulation was seen after 4 hours in the liver, kidneys, lungs, brain and spleen, whereas in the heart the maximum occurred after 8 hours. In the liver metallothionein mRNA was increased 14.5-fold over the control and in the kidneys it was increased by a factor of 9.2. A significant increase was also seen in the lungs, where it was 10 fold. To determine whether the increase is due to new transcription of the metallothionein gene, animals were pretreated with actinomycin D (1.0 mg/kg body weight) before receiving D-penicillamine. Actinomycin D prevented some of the D penicillamine-induced increase in metallothionein mRNA, indicating that the drug, to some extent, acts at the transcriptional level. Regulation of metallothionein gene expression may play an important role in the molecular mechanisms involved in the clinical action of D-penicillamine in rheumatoid arthritis. PMID- 2980794 TI - Review of publications. PMID- 2980793 TI - Zinc, copper, iron and lead status in humans following sixty days of selenium supplementation. PMID- 2980795 TI - Zinc status in pregnancy and the occurrence of anencephaly in Turkey. AB - Serum and hair zinc concentrations were measured in samples taken from 29 mothers with neural tube defects (NTD) (mainly anencephaly) at delivery. The control group consisted of 20 healthy mothers with normal offspring and 40 nonpregnant women 18 to 34 years of age, from middle income backgrounds. The mean maternal serum and hair zinc concentrations in the NTD group were lower than those of control mothers and the nonpregnant women. The zinc levels in the blood (plasma, red blood cells) and hair of 8 newborn infants with NTD (4 being anencephalic) were compared with those of normal newborn infants. The mean zinc concentration in the hair of malformed babies (250.4 +/- 85.2 micrograms/g) was significantly higher than that of normal infants (193.4 +/- 39.2 micrograms/g) (p less than 0.05) while the mean plasma zinc concentration was significantly lower (59.48 +/- 9.18 micrograms/dl compared with 68.75 +/- 10.89 micrograms/dl) (p less than 0.01). No difference was found between the mean values of red blood cell zinc levels for the two groups of infants. Maternal zinc deficiency was thought to be one of the factors responsible for NTD in Turkey. PMID- 2980796 TI - Selenium, zinc, copper and cadmium concentration in livers and kidneys of people exposed to environmental cadmium. AB - Selenium (Se), Cadmium (Cd), Zinc (Zn) and Copper (Cu) concentrations have been determined in the kidney cortex, medulla and liver of 21 Cd-exposed and 15 nonexposed autopsied subjects. In the kidney cortex, all four metals were significantly lower in the Cd-exposed subjects than in the nonexposed ones. In the liver Se levels were almost the same in both groups. Significant correlation coefficients between Se and the three metals were found in the kidney cortex of all the subjects. In the liver significant correlation coefficients were only seen between Se and Cu in the Cd-exposed subjects. These results indicated that the distribution of Se in humans chronically exposed to environmental Cd is different from that in humans exposed to environmental organic mercury. PMID- 2980797 TI - An apparent phosphate selenium interaction in weaner sheep. AB - The severity of selenium dependent nutritional myopathy in weaner sheep was markedly increased by the ruminal infusion of soluble phosphate (6.0 g d-1). Liver selenium concentration was also significantly lower in the sheep infused with sodium phosphate than in sheep fed the same diet and infused with sodium chloride. There were no significant differences in the selenium concentrations of other tissues examined (kidney, heart, skeletal muscle, blood). Blood glutathione peroxidase (GSH.Px) activity in the sheep affected with nutritional myopathy was considerably higher than values generally accepted to indicate a selenium deficiency. The results indicated that there was an interaction in the animal between selenium and phosphate leading to lower liver selenium concentrations. Results obtained from sheep intraruminally dosed with 75[Se]selenate suggested that an interaction between selenium and phosphate could also occur in the gastrointestinal tract. PMID- 2980799 TI - Review of publications. PMID- 2980798 TI - Study of the antagonism between zinc and various embryonic toxics on human amnion permeability. I. Zinc and cadmium. AB - The effects of Cd and Zn have been studied on the ionic permeation of the isolated human amnion, expressed by the measure of the transamniotic conductance Gt and the flux ratio (mother-fetus/fetus-mother: F1/F2). Cd has no action on Gt on the fetal side and decreases Gt on the maternal side by a screening effect. Zn acts on the two faces by a screening-binding effect. The ratio F1/F2 remains constant with Zn concentration, but decreases with Cd and becomes less than 1 at 5 mmol/l of Cd. F1/F2 decreases with the duration of the administration of Cd. Zn has the opposite effect of Cd on the ionic transfer and has a stabilizing action on the amnion. Moreover, Zn may antagonize the noxious effect of Cd on the maternal side. PMID- 2980800 TI - Investigation of metal-species (Ca, Mg, Zn, Fe, Cu, Pb, Cd, Sn) in urine by HPLC AAS. AB - Urine is separated by reversed-phase HPLC and metal-species are detected on-line by flame-AAS (Ca, Mg, Zn, Cu and Fe) or by ETAAS in fractions (Pb, Cd, Sn). With the exception of lead, which is present in urine in two forms (bound to chloride and to sulfate), all the metals are present as chlorides or chloro-complexes. The same results are found for two control urines. It is shown that addition of some drugs to urine (Ca-EDTA, tetracycline and barbiturate) alters the distribution of metal-species by formation of coordination compounds. PMID- 2980801 TI - The effect of desferrioxamine on tissue aluminum concentration and bone histology in aluminum-loaded rats with renal failure. AB - Aluminum (Al) intoxication is a condition that reponds well to treatment with desferrioxamine (DFO) in humans with renal failure. The present study deals with the effect of DFO administration on the Al concentrations of different tissues and on bone histology in rats with renal failure, loaded with Al. After the Al loading there is an important increase of Al in serum, (1,103 +/- 355 micrograms/l) (mean +/- SD), bone (116 +/- 14 mg/kg), liver 238 +/- 78 mg/kg) and muscle (8.5 +/- 4.1 mg/kg). Urinary Al excretion averages 287 +/- 68 micrograms/d. Serum biochemistry reveals renal failure and hypercalcemia (3.5 +/- 0.3 mmol/l). The rats were subsequently treated by either placebo or 150 mg DFO per week. The placebo-treated rats show a spontaneous decrease of serum Ca and Al and of liver and muscle Al content. The DFO-treated rats had, compared to the placebo-treated animals, an increase of urinary Al excretion and lower Al concentration in bone and brain. Bone histology after Al-loading shows an increase in osteoid measurement when compared with non-Al-loaded animals with renal failure: osteoid volume 25.6 +/- 13.3% versus 5.0 +/- 2.9% and osteoid index 63.6 +/- 22.1% versus 27.3 +/- 15.3% in Al-loaded and non-Al-loaded animals, respectively. In addition, the aluminon stain covered 89.7 +/- 3.8% of the bone trabeculae of Al-loaded rats. Under placebo treatment the osteoid measurements increase, reflecting a worsening of Al-induced bone disease. DFO therapy did not result in a significant change of the different measurements of bone histology, despite the decrease of aluminon staining to 67.7 +/- 13.7%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980802 TI - Calmodulin, zinc and calcium concentration in tissues of zinc- and calcium deficient rats. AB - To demonstrate the effect of zinc and/or calcium deficiency on calmodulin concentration and also the antagonistic behavior of zinc and calcium in metabolism, four groups of eight rats each were provided the following diets in equal amounts: group I, the basic diet with 63 ppm zinc and 0.88% calcium; group II, the calcium-deficient diet (0.0043% calcium); group III, the zinc-deficient diet (1.1 ppm zinc); and group IV, the combined zinc/calcium-deficient diet. The zinc-deficient diet significantly reduced the zinc concentration in serum (72%), femur (65%), pancrease (34%), muscle (23%), testis (11%) and brain (7%), and elevated the calcium concentration in testis (19%) and brain (24%), but did not affect calcium concentration in serum, pancreas and muscle. Likewise, calcium deficiency reduced the calcium concentration in serum (56%), femur (40%), pancreas (16%), muscle (43%), testis (19%) and brain (24%), and elevated the zinc concentration in femur (22%), although the zinc concentration in serum, pancreas, muscle and testis remained unaffected. Combined zinc and calcium deficiency elevated both the zinc as well as the calcium concentration in serum, femur and muscle, as compared to zinc or calcium deficiency alone. The alkaline phosphatase activity was reduced by 52% during zinc deficiency and was elevated by a factor of 2.5 during calcium deficiency. The calmodulin concentrations in muscle, testis and brain, as determined by radioimmunoassay, were elevated during both zinc as well as calcium deficiency, although this elevation could not be confirmed in every case. PMID- 2980803 TI - Average mineral and trace element content in daily adjusted menus (DAM) of French adults. AB - This article is based on data on food consumption gathered from 10,000 households. The daily energetic intake of 2,000 to 2,500 kcal (8,360 to 10,450 kJ) (beverages excluded) provides adults with less than 80% of the recommended amount of several essential minerals and trace elements. For women, beverages contribute only slightly to the total amount of mineral and trace element intake, because the major beverage sources of minerals and trace elements are alcoholic drinks (wine, beer). For men, alcoholic drinks could provide an additional amount of these elements. Consequently, it seems the risk of deficiency appears in adults, with women particularly concerned by this problem. The present study demonstrates that the intake of the following elements might be insufficient in women: fluorine, manganese, iron, copper, zinc and magnesium. PMID- 2980804 TI - Nutritional copper intoxication in three German infants with severe liver cell damage (features of Indian childhood cirrhosis). AB - Three of the four children of two unrelated German families fell ill in the first year of life with severe hepatopathy leading to death in two of the children so far, after a progressive clinical course and severe hepatic failure. Laboratory and morphological investigations revealed a high concentration of copper in the liver and to a lesser degree in the kidneys and other organs. The liver architecture was severely altered by micronodular cirrhosis with toxic liver cell damage similar to that found in Indian childhood cirrhosis. Epidemiologically, copper intoxication of the drinking water was verified. The drinking water was obtained from wells via copper pipes. The copper content of the drinking water went as high as 3400 micrograms/l in the two families while water taken directly from the well showed normal content but had lowered pH (6.2). Both parents were clinically healthy, as was an older son who had not been exposed to copper intoxication during the first nine months of his life. Therefore, copper intoxication during the perinatal period appears to be a prerequisite for manifestation of the disease. PMID- 2980806 TI - Mercury in whole blood of persons living in a polluted region of Turkey. AB - Hg content of whole blood has been increasing due to industrial pollution. The best means for Hg analysis is the cold vapour method. An absolute sensitivity of 0.1 ng was attained with the experimental setup in our laboratory. Its applicability was checked by A-13 freeze-dried animal blood standard reference material. The method was used to measure the levels of Hg in whole blood samples taken from eight persons living in a polluted region. Hg contents of the whole blood samples were found to lie within the range 11.66-30.83 micrograms/mL. The variables affecting the analysis, such as reducing agent type and amount, reaction time, carrier gas flow rate, reaction vessel volume, and instrumental parameters were studied and optimized. PMID- 2980805 TI - Relationship between atmospheric lead concentration and blood lead level in Turin (Italy). AB - On July 1, 1981 the Italian government received and implemented an E.E.C. directive dated 1978 calling for the reduction of lead in fuels from 0.6 to 0.4 g/L. The effectiveness of this legislative measure is evaluated here by comparison of data on atmospheric lead levels in the city of Turin, and blood lead levels in A.V.I.S. (Italian Association of Blood Donors) donors in Turin and the countryside (control group), before (1977) and after (1986, 1987) the enforcement of the law. The results show that the implementation of the law has contributed to a decrease in the atmospheric lead levels. The analysis of blood lead concentration revealed an evident biological response to this environmental change: there was a decrease in blood lead level between 1977 and 1987, in both the countryside (control group) and, to a lesser extent, in the city. PMID- 2980807 TI - Is the major part of the periodic system really inessential for life? PMID- 2980808 TI - A method for determination of zinc in mononuclear leucocytes. AB - A method for the determination of the concentration of zinc in a well characterized fraction of mononuclear leucocytes from human blood is described. Leucocytes were separated from whole blood by use of the one-step sodium metrizoate/Ficoll procedure. The cell suspensions obtained by the separation procedure had the following composition (mean, range) (N = 15): 82% (70-85%) lymphocytes, 15% (10-15%) monocytes and 4% (2-10%) neutrophils. The ratio of platelets to leucocytes was 1:1 (1:1-1.5). Cell pellets were sonicated before zinc analysis by flame atomic absorption spectrophotometry. 10 healthy volunteers (5 males and 5 females) aged 21-45 years, median age 33 years, were studied. The concentration of zinc in mononuclear leucocytes was 1.14 +/- 0.14 mumol/10(10) cells or 156 +/- 8 mumol/g protein (mean +/- SD). PMID- 2980809 TI - Retention of lead in growing rats with varying dietary lead supplements. AB - 144 female rats with a beginning weight of 35 +/- 3 g were fed for 23 days a half synthetic diet with a lead content that varied over 18 concentrations (0.025 to 600 mg/kg). Absolute lead retention was calculated as total body concentration of lead in animals at the end of the experiment minus the average lead concentration in carcasses of reference animals that had been sacrificed at the beginning of the experiment. The relative retention was calculated with regard to lead intake. Absolute lead retention increased in association with a lead supply of 0.225 mg per kg diet, but decreased thereafter. Not until a lead concentration of 5 mg per kg diet was reached did lead retention rise again. Relative lead retention decreased exponentially with an increasing lead supply and finally approached a constant value. Live mass development and feed intake were not affected by an increase in lead supply. Altogether, these results demonstrate that lead metabolism is homeostatically regulated. PMID- 2980810 TI - Effect of dietary lead intake on lead retention in adult rats. AB - 132 female rats with a live weight of 177 to 224 g were provided over a period of 17 days a half-synthetic diet with a lead concentration of 0.4 mg/kg. Then 120 of the animals were divided into 15 treatment groups fed with a diet different in Pb content (0.02, 0.04, 0.08, 0.12, 0.22, 0.42, 0.62, 0.82, 1, 5, 10, 50, 100, 400 and 800 mg Pb2+ per kg) over a period of 29 days. The feed was rationed so as to provide the minimum energy maintenance requirement. Pb retention dependent on different lead supplies was determined by subtracting the Pb contents of carcasses of 12 reference animals sacrificed at the beginning of the experiment from the Pb content of carcasses of rats at the end of the experiment. Up to a Pb concentration of 0.42 mg/kg in the diet Pb retention fluctuated around zero. Then the retention was slightly positive up to 5 mg/kg Pb in the diet. Up to 10 mg/kg Pb in the diet Pb retention increased sharply. Concentrations of iron, copper, zinc, and manganese in carcasses were affected only slightly by the varying concentration of lead in the diet. PMID- 2980811 TI - Serum selenium concentration in maternal and umbilical cord blood. Relation to course and outcome of pregnancy. AB - The present knowledge of the role of selenium in human fetal and neonatal development is sparse. In this study we measured serum selenium concentrations in maternal and umbilical cord blood from 500 Danish mothers at delivery, looking for a relationship between various maternal and fetal complications and selenium values. In mothers with uncomplicated pregnancies and deliveries serum selenium concentrations were 0.84 +/- 0.19 mumol/l (mean +/- SD), whereas in cord blood from full-term babies born adequate for gestational age and with no malformations serum selenium concentrations were 0.60 +/- 0.11 mumol/l (mean +/- SD). Mothers who received mineral tablets (other than iron preparations) during pregnancy had significantly higher serum selenium values than unsupplemented mothers (0.92 versus 0.70 mumol/l) (p less than 0.001). Infants of mineral-supplemented mothers also had significantly higher serum selenium values than infants of unsupplemented mothers (0.62 versus 0.58 mumol/l) (p less than 0.001). Preterm infants had significantly lower serum selenium concentrations than full term infants (0.54 versus 0.60 mumol/l) (p = 0.01), whereas infants small for gestational age did not differ in serum selenium concentrations from infants adequate for gestational age. Serum selenium concentrations in malformed infants tended to be lower than in normal infants (0.55 versus 0.60 mumol/l) (p = 0.09), but the difference was not statistically significant, probably due to the small number. Serum selenium concentrations in mothers with various complications during pregnancy and delivery did not differ from values in mothers with uncomplicated pregnancies and deliveries. PMID- 2980812 TI - Trace element studies on whole human cerebrospinal fluid with external beam PIXE. AB - External beam PIXE analysis with a 2.4 MeV proton beam was used to determine the concentrations of K, Ca, Fe, Cu, Zn and Br in cerebrospinal fluid from patients having various disorders. The obtained total concentration ranges K 34,000 1,079,000, Ca 5300-81,300, Fe 40-1030, Cu 20-1650, Zn 15-1250 and Br 400-43,000 micrograms/kg are compared with the values given in the literature. In certain patients there were very high CSF bromine levels, but this was shown to be the result of taking medications presented as bromide salts. The possibility of using the method in clinical practice for CSF analysis is considered. The new method of preparing self-supporting films of the samples was used. This method was further optimized by investigating in detail the use of EDTA as a homogenizer. PMID- 2980813 TI - Age-related lead increase in human permanent teeth demonstrated by energy dispersive X-ray fluorescence. AB - The combined use of a systematic sampling procedure with energy dispersive X-ray fluorescence on non-erupted and erupted permanent premolars and molars in patients aged 9 to 64 years, who have lived all their lives in Strasbourg, showed a higher total lead content in dentine compared to enamel, with an approximate ratio of 2:1. In addition, a lead increase was observed in both tissues with age. The lead increase in dentine was more significant and the highest lead values were noted in the inner pulpal dentine layer of the oldest age group. When compared to similar lead studies made on permanent teeth in urban areas, it can be concluded that the lead content in the teeth of the inhabitants of Strasbourg can be considered as low. PMID- 2980814 TI - Skin selenium content measured by hydride generation and atomic absorption spectroscopy. AB - The concentration of selenium in skin was measured using hydride generation and atomic absorption spectroscopy. The method used in this study was shown to be both accurate and precise, with mean recoveries of 92%-98% of added selenium and a within run relative standard deviation of better than 7% at a selenium concentration of less than 12 nmol/g dry tissue. The method is suitable for rapid analysis of large numbers of samples. PMID- 2980815 TI - Review of publications. PMID- 2980816 TI - The in vitro effect of lithium on growth and adherence of Streptococcus mutans 6715. AB - The in vitro effect of lithium (Li) on growth and adherence of Streptococcus mutans 6715 in the presence and absence of sucrose was examined by means of optical density and transmission electron microscopy. In the non-sucrose assay, bacterial growth was depressed and adherence was elevated between concentrations of 1.44-11.52 mmol/L Li. At 7.20 mmol/L Li extracellular material was present on the bacterial wall. Using 11.52-28.08 mmol/L Li resulted in a perfect negative correlation between the increasing concentrations of Li and bacterial growth and adherence (rs = -0.9 and rs = -1, respectively) in both assays. This paper addresses the dichotomy between the low and high Li concentrations regarding the two bacterial parameters studied, as well as their possibly related cariogenic and cariostatic clinical relevance. PMID- 2980817 TI - Correlation between urinary excretion of Tamm-Horsfall protein and sodium chloride in endemic nephropathy. AB - Excretion of Tamm-Horsfall protein and of certain electrolytes was measured in subjects living in an area of endemic nephropathy, in subjects living in a control area, and in patients with other types of chronic renal disease. A significantly higher excretion of Tamm-Horsfall protein and of sodium chloride was observed in the diseased, suspect and subjects "at risk", as compared to the control subjects and patients suffering from other types of chronic renal disease. A positive linear correlation between urinary excretion of Tamm-Horsfall protein and of sodium chloride was found in all the study groups of subjects from the endemic and control areas as well as in patients with other renal diseases. PMID- 2980818 TI - A stable tracer technique for calcium metabolism studies. AB - A tracer technique using the stable isotope calcium-48 was developed to facilitate studies of human calcium metabolism. Small samples of 1 ml serum were used. Calcium levels were determined with good precision using neutron-activation analysis after pre-concentration of the serum calcium by oxalate precipitation. The method has sufficient sensitivity to detect induced dilutions of the tracer by administering other less costly isotopes of calcium. The ability to utilize small volumes of sample is of direct benefit to paediatric metabolism research. The favourable results obtained for two calcium loading tests demonstrate the applicability of the technique to intestinal absorption studies. PMID- 2980819 TI - Concentration changes of zinc, copper and iron in serum of chronic chagasic myocardiopathic patients. AB - Zinc, iron and copper content in serum of 75 myocardiopathic chagasic patients together with that in 29 healthy controls were determined. Blood samples were taken by catheterization from the superior cava vein, coronary sinus, pulmonary artery and a peripheral artery, usually the femoral. In general, serum zinc and iron levels tend to diminish in chagasic patients, while serum copper levels tend to increase. The coronary, systemic and peripheric gradients indicate that all these metal elements tend to deposit in heart tissue. Although zinc and iron tend to be deposited also in liver and other tissues, copper is released from them into the blood stream. Further studies should determine the requirement of these metal species in heart and other tissues of experimental animals in order to obtain more conclusive information about the cause of the serum metal impairment observed here. PMID- 2980820 TI - A method of lead determination in human teeth by energy dispersive X-ray fluorescence (EDXRF). AB - A systematic sampling procedure was combined with a method of energy dispersive X ray fluorescence (EDXRF) to study lead content and its variations in human teeth. On serial ground sections made on unembedded permanent teeth of inhabitants of Strasbourg with a special diamond rotating disk, 2 series of 500 microns large punch biopsies were made systematically in 5 directions from the tooth surface to the inner pulpal dentine with a micro-punching unit. In addition, pooled fragments of enamel and dentine were made for each tooth. On each punched fragment or pooled sample, lead content was determined after dissolution in ultrapure nitric acid, on a 4 microns thick polypropylene film, and irradiation with a Siemens EDXRF prototype with direct sample excitation by a high power X ray tube with a molybdenum anode. Fluorescence was detected by a Si(Li) detector and calcium was used as an internal standard. This technique allowed a rapid, automatic, multielementary and non-destructive analysis of microsamples with good detection limits. PMID- 2980821 TI - Dose-dependent distribution of injected selenium in rat blood. Effect of previous selenium intake in drinking water. AB - The effect of an administered dose of 75Se-selenite and a previously increased selenium intake in drinking water (0.1 mg/l) on the distribution of injected selenium in rat blood was studied. In a dose range of 0.01-1.6 mg Se/kg body weight the ratio of injected selenium in blood plasma and in blood cells decreased from 3.20 in rats with increased selenium intake and 4.60 in rats without this intake, to 0.13 and 0.10, respectively. After injection of high selenite doses, 85-88% of the selenium present in the blood was localized in blood cells irrespective of increased selenium intake by drinking water. Possible relationship between accumulation of selenium in blood cells and its toxic effect in the organism is discussed. Previously increased intake of selenium had no effect on the levels of selenium in blood cells but affected significantly its plasma levels. The results indicate that the previously increased selenium intake in drinking water increases the capacity of the plasma for selenium injected in the form of selenite. PMID- 2980822 TI - Metallothionein in human liver and kidney: relationship to age, sex, diseases and tobacco and alcohol use. AB - The content of metallothionein (MT) was determined in the livers (MT-L) and the kidneys (MT-K) of 145 deceased persons. The individual values showed marked variations; average levels (arithmetic means +/- S.D.) were 154.9 +/- 151.4 mg/kg liver wet wt. and 160.5 +/- 150.4 mg/kg total kidney wet wt. In contrast to MT-L, MT-K increased with age up to a maximum around mid-life and decreased at higher ages. Neither the MT-L nor the MT-K depend on sex. The MT content significantly correlates with the macroscopic and histopathologic status of the liver. As compared to normal tissue, livers with fatty degeneration, cirrhosis and brown atrophy show MT-L values of 40%, 25% and 233% respectively. Low MT-L of 40% of the control values are also observed in individuals with a history of alcohol abuse. This decline preceded visible macroscopic pathological findings. MT-L also responds to the cause of death. At suicidal overdose of drugs the MT-L is lowered to 62% and at "mechanical" suicides it is increased to 137%. Whereas MT-K is essentially independent of the kidney status, it is significantly increased to approximately 190% in male smokers. PMID- 2980823 TI - Serum and urinary magnesium, calcium and copper levels in insulin-dependent diabetic women. AB - Eighteen insulin-dependent diabetic (IDD) and 15 healthy control women participated in a study of the relationship between glycemic control or renal function measurements and the levels of magnesium, calcium and copper in the serum and urine. In the IDD women glycosylated hemoglobin averaged 9.8 +/- 0.5% and the fasting plasma glucose was 1.89 +/- 0.19 g/L. The glomerular filtration rate, approximated from creatinine clearance, averaged 125 +/- 16 ml/min in the IDD women in comparison to 82 +/- 4 ml/min in the controls. IDD women had lower serum (p less than 0.001) and higher urinary (p less than 0.01) magnesium levels than did the controls; serum and urinary copper levels and serum ceruloplasmin levels all were higher (p less than 0.01) in the IDD women. There was no difference in either serum or urinary calcium levels between the two groups. In the IDD women serum magnesium was related to urinary glucose (r = -0.758, p less than 0.01) and to glycosylated hemoglobin (r = -0.603, p less than 0.01); none of the other measurements of serum or urinary minerals was related to measures of glucose control. Urinary calcium and copper levels, but not urinary magnesium, were correlated with the glomerular filtration rate (r = 0.476, p less than 0.05 and r = 0.554, p less than 0.05, respectively). The results of this study suggest that IDD alters the utilization of magnesium, calcium, and copper, but that these three minerals are not affected in the same way by physiological changes associated with the disease. PMID- 2980824 TI - Circadian variations in plasma zinc and cortisol in man. AB - Circadian variations of zinc and cortisol concentrations in plasma were studied in six healthy adult men. Three of them were tested over two different 24-h periods. Results were analyzed by computerizing a best-fit curve for each 24-h profile. Plasma zinc displayed a morning peak between 8.00 and 9.00 a.m. followed by an almost linear decline throughout the day with lowest values observed shortly before 6.00 p.m. A transitory increase occurred between 6.00 p.m. and 8.00 p.m. followed by a slow decrease reaching its nadir around midnight. Thereafter zinc increased steadily until 8.00 a.m. A similar profile was observed in a seventh subject who was undergoing therapeutic starvation for obesity (fifth day of the starvation period). In all subjects the time course of plasma cortisol fluctuations paralleled that of zinc. Our results confirm the existence of a circadian rhythm in plasma zinc independent of zinc intake and temporally related to the circadian rhythm of cortisol. PMID- 2980825 TI - Review of publications. PMID- 2980826 TI - Topical cyclosporin A in the treatment of corneal graft reaction. PMID- 2980827 TI - Major perils of minor surgery. PMID- 2980828 TI - Penetrating keratoplasty in aphakic and pseudophakic bullous keratopathy. PMID- 2980829 TI - Importance of HLA DR matching for corneal transplantation in high-risk cases. PMID- 2980830 TI - Quantitative parameters of a high-affinity binding site for retinoic acid in corneal endothelial cells. PMID- 2980831 TI - Serotyping of ocular Pseudomonas aeruginosa clinical isolates. PMID- 2980832 TI - The pathogenesis of contact lens-associated Pseudomonas aeruginosa corneal ulceration. II. An animal model. PMID- 2980833 TI - The measurement of mucin in human tears by high-pressure liquid chromatography. PMID- 2980834 TI - Extraction of soluble proteins reduces rejection in rabbit porcine corneal xenografts. PMID- 2980835 TI - Intracanalicular collagen implants enhance topical antibiotic bioavailability. PMID- 2980836 TI - Cryopexy of epithelial downgrowth. PMID- 2980837 TI - The effect of tuftsin in the treatment of experimental Pseudomonas keratitis. PMID- 2980839 TI - Food allergy and behavior: definitions, mechanisms and a review of the evidence. PMID- 2980838 TI - Squamous cell carcinoma of the cornea and conjunctiva following a thermal burn of the eye. PMID- 2980840 TI - Optimal methods and issues in nutrition research in the correctional setting. PMID- 2980841 TI - Prison dietary standards and the research potential for evaluating the effect of diet upon behavior. PMID- 2980842 TI - Assessment of the classroom behavior of hyperactive children. PMID- 2980843 TI - Double-blind challenge studies of behavioral and cognitive effects of sucrose aspartame ingestion in normal children. PMID- 2980844 TI - Blood-brain barrier transport of nutrients. PMID- 2980845 TI - Associations between nutrition and behavior in 5-year-old children. PMID- 2980846 TI - Diet and hyperactivity. PMID- 2980848 TI - Complex interactions affecting nutrition-behavior research. PMID- 2980847 TI - Acute effects of meals on perceptual and cognitive efficiency. PMID- 2980849 TI - Nutritional control in animal experiments. PMID- 2980850 TI - Methods for assessing the adverse effects of foods and other chemicals on animal behavior. PMID- 2980851 TI - Dietary survey data: sources and limits to interpretation. PMID- 2980852 TI - Evaluating the nutritional status of individuals: a critique of approaches. PMID- 2980853 TI - Behavior as an outcome in nutrition research. PMID- 2980854 TI - Individuality in the response to dietary constituents: some lessons from drugs. PMID- 2980855 TI - The regulatory perspective of diet-behavior relationships. PMID- 2980856 TI - Dietary pharmacology. PMID- 2980857 TI - Acute and chronic effects of protein and carbohydrate ingestion on brain tryptophan levels and serotonin synthesis. PMID- 2980858 TI - The behavioral effects of food constituents: strategies used in studies of amino acids, protein, carbohydrate and caffeine. PMID- 2980859 TI - Effect of L-tryptophan and other amino acids on sleep. PMID- 2980860 TI - Nutrients and newborn behavior: neurotransmitters as mediators? PMID- 2980861 TI - Psychiatric aspects of the relationship between eating and mood. PMID- 2980862 TI - Diet, crime and delinquency: a critique. PMID- 2980863 TI - Reactive hypoglycemic tendency among habitually violent offenders. PMID- 2980864 TI - The resolution solution: longitudinal examination of New Year's change attempts. AB - This study prospectively tracked the self-change attempts of 200 New Year's resolvers over a 2-year period in order to more fully understand the coping determinants of maintenance and the natural history of lapses and relapses. Seventy-seven percent maintained their pledges for 1 week but only 19% for 2 years. Successful resolvers reported employing significantly more stimulus control, reinforcement, and willpower than the unsuccessful over the 2 years; social support and interpersonal strategies failed to predict success before 6 months but did so thereafter. Counterconditioning and fading were retrospectively nominated as the most efficacious coping strategies; paucity of willpower and failure of stimulus control were reported as the most hindering to maintenance. Fifty-three percent of the successful group experienced at least one slip, and the mean number of slips over the 2-year interval was 14. Slips were typically precipitated by a lack of personal control, excessive stress, and negative emotion. PMID- 2980865 TI - Assessing life stressors and social resources: applications to alcoholic patients. AB - A growing body of evidence points to the importance of life stressors and social resources in the development and course of alcoholism and other substance abuse disorders. This article describes the Life Stressors and Social Resources Inventory (LISRES), which provides an integrated assessment of life stressors and social resources in eight domains: physical health, home/neighborhood, financial, work, spouse/partner, children, extended family, and friends. The indices were developed on data obtained at two points in time 18 months apart from four demographically comparable groups: alcoholic patients, depressed patients, arthritic patients, and non-problem-drinking adults. As expected, alcoholic patients reported more acute and chronic stressors and fewer social resources than did non-problem-drinking adults. More important, the indices were predictively related to changes in alcohol consumption, drinking problems, depression, and self-confidence. Procedures such as the LISRES have some potential clinical and research applications and may be helpful in examining the process of recovery and relapse in substance abuse disorders. PMID- 2980866 TI - Effect of naloxone on cigarette smoking. AB - Several recent studies have tried to link endogenous opioid peptides (endorphins) with smoking reinforcement. Karras and Kane (1980) showed a one-third decrease in smoking after administration of the opiate antagonist naloxone. However, Nemeth Coslett and Griffiths (1986) failed to replicate this result with use of a wide range of doses. The current study was an attempt to replicate Karras and Kane's results using 10 male chronic smokers. Each subject received naloxone (10 mg sc) or placebo in a double-blind, cross-over design (cross-over interval 6-14 days) after overnight abstinence from tobacco use. Naloxone reduced the number of cigarettes smoked by 16% in the first 1/2 hour smoking period, with a 10% reduction in carbon monoxide level 1 hour later. Naloxone had no effect on subjective and physiological variables and did not cause any signs of a tobacco withdrawal syndrome. These results are consistent with those of Karras and Kane, and suggest a role for endorphins in smoking reinforcement. Inconsistent results across studies may be due to methodological factors, such as differences in smoking deprivation time and postnaloxone administration testing intervals because naloxone has a short half-life. PMID- 2980867 TI - Combined behavioral and pharmacological treatment of alcoholic methadone patients. AB - Alcoholism is a prevalent and often debilitating problem among methadone maintenance patients. In this study, the efficacy of a contingency management program in the treatment of three severely alcoholic methadone patients was examined using within-subject experimental designs. The contingency management program made continuation in methadone maintenance contingent upon daily disulfiram consumption. Failure to consume disulfiram resulted in the transfer of the patient to another methadone program. This treatment resulted in clinically significant decreases in alcohol consumption and improvements in clinic attendance. PMID- 2980868 TI - A study examining the psychometric properties of the SMAST-13. AB - The psychometric properties of the Short Michigan Alcoholism Screening Test (SMAST-13), a 13-question abbreviated form of the MAST, were determined on two samples of alcoholics, with their nonalcoholic family members serving as controls. The study hypothesized that the SMAST-13 has a low specificity when used to screen family members for alcohol problems and would exhibit a low internal reliability coefficient in this population. All participants were interviewed using DSM-III criteria for alcohol abuse or dependence. Only family members who failed to meet these criteria were enrolled in the study. When a weighted cut-off score of 5 or more was used as a positive score for alcohol problems, the sensitivity was .98 in Sample A and .94 in Sample B. Utilizing the scores of the family members as controls, the specificity was .58 in Sample A and .70 in Sample B. The internal reliability of the SMAST-13, using Chronbach's alpha on the total sample, produced a reliability coefficient of r = .57 in Sample A and r = .62 in Sample B. To determine if the specificity of the SMAST-13 could be improved, the weighted cut-off score for a positive response was raised from 5 to 10. In Sample A, the specificity changed from .58 to .90 and the sensitivity dropped slightly from .98 to .92. In sample B, the specificity increased from .70 to .95; the sensitivity decreased from .95 to .85. Suggestions for changing the wording of two questions which explain the majority of the false positives are discussed. Epidemiological studies which have used the SMAST-13 to estimate the prevalence of alcohol problems in family members of alcoholics may need to reconsider their methods of classifying alcoholics. PMID- 2980869 TI - Assessment of disordered eating in females: instrument validation. AB - The purpose of this study was to validate an instrument capable of assessing 10 dimensions related to disordered eating and weight control behaviors. All subjects were female and ranged in age from 14 to 40. Of these females 68 were bulimics, 12 were compulsive eaters, and 24 eating disorders. The Disordered Eating and Weight Control Instrument (DEWCI) was administered. Differences between the groups were investigated by constructing three dummy variables contrasting the groups, correlating the scales with the dummy variables, and determining which correlations were significant. Eight of the DEWCI scales significantly (p less than .005) differentiated between the three groups. This instrument may therefore have utility for use with female populations. PMID- 2980870 TI - The effect of two nutritional software programs used as adjuncts to the behavioral treatment of obesity. AB - The need to provide nutritional information in the behavioral treatment of obesity is now generally recognized. However, the comparative efficacy of various delivery modes remains to be demonstrated. Two commercial software packages (The Eating Machine and EATS) were embedded in Ferguson's (1975) prototypical behavioral program and contrasted with the Ferguson approach deployed alone. Assessments of weight, nutritional knowledge, eating behavior, and related cognitive variables were made at pretest, posttest, 1-month follow-up and 6-month follow-up occasions. No incremental effects attributable to the software programs appeared. PMID- 2980871 TI - Eating disorders and substance abuse in women: a comparative study of MMPI patterns. AB - Psychopathology was compared in women with eating disorders, women with alcohol or drug problems, and women with both an eating disorder and an alcohol or drug problem. Overall psychopathology, as measured by the Minnesota Multiphasic Personality Inventory (MMPI), was greatest in the group with both an eating disorder and substance abuse problems. In general, the women with eating disorders resembled the substance abusers in terms of rebellious and antisocial behavior but differed from the substance abusers in being less hyperactive and experiencing more psychic distress. The differences that were found between eating-disordered women with and without substance abuse problems suggest that treatment interventions may need to be modified if the eating disorder patient has substance abuse problems as well. PMID- 2980872 TI - Self-efficacy, standards, and abstinence violation: a comparison between newly sober and long-term sober alcoholics. AB - Recent theory and empirical data suggest that self-efficacy plays an important role in resistance to relapse for substance abusers. Another key in the relapse process, according to Marlatt and Gordon (1985), is the abstinence violation effect, which comprises self-attribution for failure and affective reaction to violation of self-imposed standards. The combination of unrealistically high standards and low self-efficacy for following those standards may potentiate the risk for relapse. A 25-item questionnaire designed to assess self-efficacy and standards was administered to alcoholics newly admitted to an inpatient treatment program and alcoholics who had been sober for at least 1 year. The groups did not differ with regard to having high standards, but the successfully abstinent alcoholics had significantly higher self-efficacy expectations than the newly sober alcoholics. These results suggest that treatment programs may need to include interventions which decrease unrealistic standards as well as those designed to increase self-efficacy expectations. PMID- 2980873 TI - The relationship between ethanol intake and DSM-III alcohol use disorders: a cross-perspective analysis. AB - The purpose of this investigation was to quantify the relationship between ethanol consumption and DSM-III alcohol use disorders using mathematical modeling techniques that allowed for the simultaneous control of several extraneous factors and the assessment of potential interaction. Although gender, education, ethnicity, and marital status were not identified as actual confounders in the logistic regression model, the ethanol intake-dependence association was found to be stronger among younger adults than in the later stages of life. Age was an influential confounder of the ethanol intake-abuse relationship, but the magnitude of the association was generally weaker for abuse than for dependence. Separate analyses in which the abuse and dependence criteria served as outcome measures qualified the interpretation of the overall ethanol intake-disorder associations. Implications of these results are discussed in terms of age differences in exposure and context of consumption, differential interpretation of withdrawal symptoms, and the relationship between abuse and dependence. The need for future research to refine our descriptions of risk of alcohol use disorders in relation to levels of intake is highlighted. PMID- 2980874 TI - Disease staging. Applications for utilization review and quality assurance. PMID- 2980875 TI - An objective quality assurance method for the solo surgical pathologist. PMID- 2980876 TI - ACURP--the beginning. PMID- 2980877 TI - Quality assurance in our health care system. PMID- 2980878 TI - Physicians and the Peer Review Organization review. PMID- 2980879 TI - An emerging alternate level of health care and the hospital role. PMID- 2980881 TI - What is "appropriate" utilization of diagnostic tests? Technical performance, diagnostic performance, and clinical utility. PMID- 2980880 TI - Can medical ethics survive medical economics? PMID- 2980882 TI - An objective quality assurance method for the solo cytopathologist. PMID- 2980883 TI - Peer Review Organizations: the future agenda. A report of the American Medical Peer Review Association's Task Force on Peer Review Organization Implementation. PMID- 2980884 TI - Hippocratic medicine: the ambiguous legacy of ancient Greece. PMID- 2980885 TI - Medical staff bylaws: the sensitive matter of removal of officers. PMID- 2980886 TI - Case study. Part I: The sociopsychological dynamics of implementing quality assurance in a mentally retarded and developmentally disabled setting. PMID- 2980887 TI - On the need for a physician residency in quality assurance. PMID- 2980888 TI - Applying utilization review findings in medical staff appointment and reappointment decisions. PMID- 2980889 TI - Ambulatory surgery. Evolution, organization, cost efficiency. PMID- 2980890 TI - A quality assurance program for a large health maintenance organization. PMID- 2980891 TI - Preventing disease, creating society. PMID- 2980892 TI - Case study. Part II: The analysis of the sociopsychological dynamics of implementing quality assurance in a mentally retarded and developmentally disabled setting. PMID- 2980893 TI - Ethics and the health care revolution. PMID- 2980894 TI - Medical decisions: ethical and economic dimensions. PMID- 2980895 TI - Health care ethics and health care economics. PMID- 2980896 TI - Whose life and whose death. PMID- 2980897 TI - Some legal considerations about making advance arrangements for care and treatment during a terminal illness. PMID- 2980898 TI - Case study. S. J., 76-year-old male--C.V.A. PMID- 2980899 TI - Rationing--an ethical issue? PMID- 2980900 TI - Toward a definition of roles for physicians in quality assurance. PMID- 2980901 TI - Physician practice patterns: why and how we should use them. PMID- 2980902 TI - Quality assurance concerns in joint ventures. PMID- 2980903 TI - The need for an informed credentialing program. PMID- 2980904 TI - Malpractice in the emergency room: a critical review of undiagnosed appendicitis cases and legal actions. PMID- 2980905 TI - Appropriate Utilization of Resources Program. PMID- 2980906 TI - Combined plan to reduce readmissions. PMID- 2980907 TI - Case study. The use of DRG data for discharge planning. PMID- 2980908 TI - Medication costs in different provider settings. AB - Comparison was made of the cost of medications between Shared Health Facilities (SHFs) or Medicaid Mills and a Neighborhood Health Center (NHC) for nine conditions in fields of adult medicine, pediatrics, and gynecology. A total of 10 cases from SHF reviews were matched by diagnosis, age, and length of time under care with those in the NHC. For otitis media and pharyngitis in children and questionable urinary tract infection and vaginitis in adult women, the average costs were significantly higher in the SHFs. Average costs for family planning services and vaginal bleeding were higher in the SHFs but not significantly so. Medication costs for children with asthma and adults with bronchitis and hypertension were approximately the same in both settings. The reasons for higher costs included greater use of more expensive antibiotics, concurrent use of decongestants and antihistamines for infectious conditions of childhood, and dispensing of medications on "shot-gun" basis without adequate diagnostic studies. The most striking difference was the additional average cost of $798 for hospitalization of the SHF patients with vaginal bleeding when D & C and surgery were performed. The NHC women had no hospitalizations as the recommended procedure of endocervical biopsy in the office was Center policy. Other quality inferences noted in the NHC, but not in SHFs, were routine performance of throat cultures for pharyngitis, wet smears for vaginitis, and deferral of treatment until cultures were received for urinary tract infection. The small number of cases for each condition and the large intersample variability were limiting factors in this study, but the findings do suggest that higher standards of care contribute to lower therapy costs. PMID- 2980909 TI - Cost containment, data collection, and "buying right". PMID- 2980910 TI - Health care technology and quality of care. AB - The increasing costs and complexity of technologic advances in diagnosis and treatment have been accompanied by other important issues. They are often moral or ethical in nature; they include the public's desire and determination to have access to these "high-tech" advances; and the quality and equity with which those advances are apportioned and applied must be addressed. Seven criteria that can be applied to technology assessment are identified as is a process for that assessment. Together, these procedures can provide valuable information and assistance to those who make decisions about health benefits coverage--both in the public and the private sectors. PMID- 2980911 TI - Cost containment and child health. AB - Children, as consumers of health resources, have special developmental, psychological, and medical needs different from those of adults. Thus, cost containment efforts can affect children differently. Data related to insurance benefits changes, intensified market forces, and reductions in federal funding are cited. Their analysis focuses on the importance of accountability in applying cost constraints to services that can have a significant effect upon the health and well-being of one quarter of the next generation. PMID- 2980912 TI - American Medical Association update on quality assurance. PMID- 2980913 TI - Case study. Case: L. R., 53-year-old woman--glaucoma. PMID- 2980914 TI - Quality assurance principles for the laboratory--a brief overview. AB - Under the Omnibus Budget Reconciliation Act of 1986, the United States Congress has called for a study of the quality of care of Medicare beneficiaries and a strategy to assure it. The quality of laboratory services is an important component of that assessment and assurance, and hospitals need a methodology for it. The Guidelines for Quality Assessment of the Council on Medical Service of the American Medical Association can be used as a basis for a model that links structure, process, and outcome criteria to the interplay of people, equipment, and systems that comprise the laboratory. This can then support the development of a monitoring and evaluation program for its services. PMID- 2980915 TI - Quality review in the Peer Review Organization Program. AB - Several major, national forces have created the mandate of the Peer Review Organizations (PROs) for the evaluation of the quality of care received by Medicare patients. The shift of the Medicare Program to prospective payment to hospitals has caused concern that this change in financial incentives may have adverse effects on quality. An analysis and review of the experience of the California PRO (California Medical Review, Inc.)--including the application of sanctions--suggests that corrective actions have been successful in addressing quality deficiencies. However, the relationship between those deficiencies and the prospective payment mechanism remains unclear. PMID- 2980917 TI - Case study: quality community health--Hong Kong. PMID- 2980916 TI - HMO Quality of Care Program. PMID- 2980918 TI - The nuts and bolts of utilization review. PMID- 2980920 TI - Toward an educational model for physician fellowships and residencies in quality assurance and utilization review. PMID- 2980919 TI - The cost of quality assurance: an exploratory study. PMID- 2980922 TI - Legal aspects of the medical staff peer review process. The Health Care Quality Improvement Act of 1986--boon or bane? PMID- 2980921 TI - State health data systems: the Pennsylvania story. PMID- 2980923 TI - Case study. 27-yr-old man pacemaker candidate. PMID- 2980924 TI - Medical care value purchasing: medicolegal promises, pitfalls, and progress. PMID- 2980925 TI - Ethical issues associated with peer review in Medicare. PMID- 2980926 TI - The price of miracles. PMID- 2980927 TI - Quality issues in discharge planning. PMID- 2980928 TI - Quality assurance/utilization review as a practice option: the influence of a residency program. PMID- 2980929 TI - Uses of the peer review organization Medicare database. PMID- 2980930 TI - Case study. Admission denial, 7 acute care days (LOS 7 days). PMID- 2980931 TI - Legal aspects of peer review. Patrick v Burget in the U.S. Supreme Court: its impact on peer review. PMID- 2980932 TI - Quality initiatives. The role of medical licensing and disciplinary boards. PMID- 2980934 TI - Assuring quality in proprietary managed care systems. PMID- 2980933 TI - Impact of the diagnostic-related group prospective payment system on the outcome and quality of care of elderly patients hospitalized with pneumonia. PMID- 2980935 TI - Evaluating the effectiveness of the utilization management program. PMID- 2980936 TI - Peer review and you. PMID- 2980937 TI - Drug utilization management includes physician prescribing profiles. PMID- 2980939 TI - Legal aspects of the medical staff peer review process. Medicolegal issues arising in the emerging mandated health data disclosure era. PMID- 2980938 TI - A successful approach to the start up of a mental health case management program. PMID- 2980940 TI - Utilization review case study 2: dilemmas in utilization review. PMID- 2980941 TI - Tracking quality assurance activity. A PC-based computerized system. PMID- 2980942 TI - The timeliness of discharge planning. PMID- 2980943 TI - The Inspector General. Sanctions and quality of care. PMID- 2980945 TI - Legal aspects of the medical staff peer review process. Legally permissible uses of mandated health data disclosure. PMID- 2980944 TI - Quality and cost-efficiency. The evidence for channeling. PMID- 2980946 TI - Microangiopathic retinopathy in experimental diabetic monkeys. PMID- 2980947 TI - Suppressor deletion therapy. Selective elimination of T suppressor cells using a hematoporphyrin-conjugated monoclonal antibody. PMID- 2980948 TI - Hemodynamic variables and the incidence of prebypass ischemia during sufentanil/O2/pancuronium anesthesia in patients undergoing coronary artery surgery. AB - It has been suggested that sufentanil is a superior anesthetic to fentanyl for patients undergoing myocardial revascularization. This study was performed to determine the incidence of prebypass myocardial ischemia using sufentanil, 20 micrograms/kg for patients undergoing coronary artery bypass grafting (CABG). Twenty-seven patients with normal left ventricular function, scheduled for elective CABG, were studied. Anesthesia was induced with sufentanil, 10 micrograms/kg; and pancuronium, 0.1 mg/kg, was given for muscle relaxation. Further increments of sufentanil, 5 micrograms/kg, were given before skin incision and sternotomy. All patients had ECG leads V6 and V9 recorded continuously with a Holter monitor from arrival in the operating room until the start of bypass. Hemodynamic profiles were recorded at specific intervals in the prebypass period. Seven patients (25.9%) developed prebypass myocardial ischemia, three at intubation, and three at aortic dissection. Fourteen patients developed hypertension and 14 had hypotension, defined as increases or decreases greater than 20% from baseline values, respectively. Only five patients had neither hypotension nor hypertension. Three patients (11.1%) had perioperative myocardial infarctions, two of whom had prebypass myocardial ischemia. It is concluded that the incidence of prebypass ischemia with sufentanil anesthesia approximates 26%, which is similar to other studies using sensitive ECG monitoring techniques for the detection of ST segment changes. Hemodynamic instability, in the form of bradycardia, hypertension and hypotension, but not tachycardia, may have contributed to the incidence of prebypass ischemia. PMID- 2980949 TI - A retrospective analysis of fentanyl and sufentanil for cardiac transplantation. AB - Drug requirements, hemodynamic responses, and fluid balance were retrospectively studied in 65 male patients undergoing cardiac transplantation. Twenty patients received fentanyl-oxygen (88 +/- 7 micrograms/kg), and 45 patients received sufentanil-oxygen (20 +/- 1 micrograms/kg) as the primary anesthetic technique. The left ventricular ejection fraction averaged 15% and 18%, respectively. Hemodynamic stability was maintained during induction, bypass, and postbypass periods with both drugs. Cardiopulmonary bypass (CPB) was terminated with isoproterenol support for all patients. Additional inotropic support was required for 65% of fentanyl- and 56% of sufentanil-treated patients. Pacemaker support was required less frequently for sufentanil- than fentanyl-treated patients (P less than .01). Vasodilator therapy was required after CPB for patients receiving sufentanil despite the fact that higher-than-anticipated doses of sufentanil were administered during and after CPB. Fluid requirements were similar for both groups. It is concluded that both high-dose fentanyl and sufentanil anesthesia can provide clinically satisfactory anesthesia for cardiac transplantation. PMID- 2980950 TI - Multifactorial index of cardiac risk in noncardiac surgery: ten-year status report. PMID- 2980951 TI - Plasma atrial natriuretic peptide concentrations during induction of anesthesia and acute volume loading in patients undergoing cardiac surgery. AB - Induction of anesthesia with fentanyl for coronary artery bypass grafting decreased (P less than .05) plasma atrial natriuretic peptide (ANP) concentrations from awake values in twelve patients. During a steady state of anesthesia before surgery, isotonic saline solution (10 mL/kg) was infused simultaneously with the elevation of the lower extremities in six patients, while six subjects served as controls receiving no volume loading and having no leg raising. The ANP levels returned to the awake values in the volume-loaded patients, while plasma ANP remained at anesthetized baseline levels in the control subjects (P less than .01 between the groups). Ten minutes after the end of the loading procedure, plasma ANP had begun to decrease again towards the postinduction level in the loaded group, but a significant (P less than .05) difference was still observed between the groups. These changes in ANP levels paralleled those of cardiac filling pressures. In conclusion, the results suggest that the degree of distention of the atria regulates the secretion of atrial natriuretic peptide into the circulation in patients anesthetized with fentanyl. PMID- 2980952 TI - Catecholamine responses to anesthetic induction with fentanyl and sufentanil. AB - In a randomized study, the authors examined the changes in plasma epinephrine and norepinephrine concentrations associated with induction of anesthesia and surgery in 33 patients with good ventricular function undergoing elective coronary artery surgery. After premedication with morphine and scopolamine, patients received either fentanyl, 100 micrograms/kg (n = 16), or sufentanil, 15 micrograms/kg, (n = 17), intravenously (IV), over 10 minutes to induce anesthesia. Metocurine, 0.42 mg/kg, IV, produced muscle relaxation. Arterial blood for plasma catecholamine determinations was drawn prior to induction, every two minutes throughout induction, one minute following endotracheal intubation, and one minute after sternotomy. Plasma epinephrine concentration was unchanged with either induction agent. Plasma norepinephrine concentration increased significantly after administration of either narcotic, peaked between six and ten minutes into induction, and returned to the preinduction value after intubation. Induction related changes in arterial pressure and pulmonary capillary wedge pressure were significantly correlated with changes in the logarithm of plasma norepinephrine concentration. Similar degrees of endogenous norepinephrine release appear to accompany induction with equipotent doses of fentanyl and sufentanil in patients premedicated with morphine and scopolamine. Norepinephrine release may influence the hemodynamic response to induction with narcotics. PMID- 2980953 TI - Evolution of narcotic anesthesia for cardiac surgery. PMID- 2980954 TI - Comparison of sufentanil-oxygen and fentanyl-oxygen anesthesia for mitral and aortic valvular surgery. AB - The cardiovascular responses, speed of anesthetic induction, incidence of chest wall rigidity, need for anesthetic supplements (phentolamine, N2O, and nitroprusside) to control intraoperative hypertension, and speed of postoperative recovery were measured and compared in 44 patients undergoing aortic and mitral valvular replacement with fentanyl-O2 or sufentanil-O2 anesthesia. After a lorazepamatropine premedication and pancuronium pretreatment, fentanyl was administered intravenously at a rate of 400 micrograms/min and sufentanil at 200 micrograms/min until patients were unconscious; at this time they were given succinylcholine and their tracheas were intubated. After intubation, an amount of fentanyl or sufentanil equal to the dose producing unconsciousness was infused over the next 30 minutes, at which time the operation began. Additional fentanyl or sufentanil was given whenever systolic arterial blood pressure (SBP) increased more than 15% over preanesthetic values. When three successive supplemental doses of the narcotic failed to effectively decrease SBP, phentolamine was used to control pressure before and during bypass; after bypass, N2O (25% to 50%) or, if N2O was ineffective, nitroprusside was used. Average time of induction was 3.4 +/ 0.3 for fentanyl and 1.0 +/- 0.2 min (mean +/- SD) for sufentanil. Chest wall rigidity occurred in 36% of patients in both groups. Total doses of fentanyl and sufentanil required for the entire operation were 113 +/- 11 and 9.0 +/- 0.4 micrograms/kg (mean +/- SD), respectively. Heart rate, cardiac output, and mean right atrial pressure remained unchanged throughout the study in both groups. Mean arterial blood pressure (MBP) and SBP were significantly decreased during induction and after intubation in patients receiving sufentanil, but not fentanyl.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980955 TI - Low ventilatory and anesthetic drug requirements during myocardial revascularization in a hypothyroid patient. PMID- 2980957 TI - Population genetic studies on the PGM1 isoenzyme system in Szeged and its surroundings. PMID- 2980956 TI - Meta-analysis of randomized controlled trials as a method of estimating rare complications of non-steroidal anti-inflammatory drug therapy. AB - The design of randomized controlled trials to assess the efficacy of pharmacological measures for the prevention of the gastrointestinal side-effects of anti-inflammatory drugs requires an accurate estimate of excess risk under controlled conditions. Photocopies of 952 randomized controlled trial publications were obtained after scanning titles and abstracts of a MEDLINE computer search, 427 were excluded for obvious reasons, and 525 were again photocopied after obliterating source and results. Selection criteria were: the presence of a non-anti-inflammatory drug control group; at least 4 days of therapy; at least 3 days without anti-inflammatory drugs before randomization; no complicating background drugs; mention of side-effects; and a clear differentiation of gastrointestinal complications. Observer error, with two independent readings, for inclusion suitability in the study was 19% for Methods and 9% for Results. For the 44 aspirin trials, the mean therapy duration was 357 days; the unweighted rate difference between therapy and control groups ( +/- 1 S.E.M.) for ulcer was 0.006 +/- 0.003, for gross haemorrhage 0.006 +/- 0.002 and for unspecified gastric symptoms 0.03 +/- 0.01. In 123 non-aspirin non-steroidal anti-inflammatory drug (NA-NSAID) trials, the mean duration was 67 days; the unweighted rate difference for ulcer was 0.0005 +/- 0.0003, for gross haemorrhage 0.007 +/- 0.004 and for unspecified gastric symptoms 0.02 +/- 0.005. Risk differences were also pooled using the DerSimonian and Laird method, which weights studies inversely according to variance. Using this method, only the unspecified gastric symptoms for non-aspirin non-steroidal anti-inflammatory drugs (NA-NSAIDs) and the haemorrhage for aspirin were found to be statistically significant. Longer studies have higher risk differences. Randomized control trials to determine prophylactic efficacy against haemorrhage (that is, to demonstrate a reduction of ulcer rate in the therapy group to the rate of controls) would require 190 patients in each group for NA-NSAIDs in studies of 2 6 months; 950 subjects would be needed to detect a 50% reduction. Randomized control trials to determine a reduction in ulcer rate to that of controls in patients on aspirin for more than 6 months would require 700 subjects in each group; 3346 subjects would be needed to detect a 50% reduction. Such studies are feasible. PMID- 2980958 TI - [Doctor-patient relationship in the process of expert testimony]. PMID- 2980959 TI - Sexual abuse of disabled youngsters. PMID- 2980960 TI - Post mortem stability of atrial natriuretic peptides. PMID- 2980961 TI - [Methodology of the evaluation and compensation for bodily injury and social injury in the common law]. PMID- 2980962 TI - Location of genes on chromosome arms in the Anopheles gambiae group of species and their correlation to linkage data for other anopheline mosquitoes. AB - The use of paracentric inversions as genetic markers in the Anopheles gambiae group of mosquitoes is described. The gene for dieldrin resistance is assigned to chromosome 2 which in turn is correlated to the previous assignment of the gene to linkage group II. The locus of the enzyme phosphoglucomutase 2 (Pgm 2) is similarly assigned to chromosome 2 and evidence is presented for possible linkage between Pgm 2 and dieldrin resistance. There was no linkage or correlation of chromosome 2 and loci of the enzymes superoxide dismutase (Sod) and octanol dehydrogenase (Odh). These genes are therefore assumed to be on chromosome 3 (linkage group III). Evidence that such gene linkage group/chromosome correlations may extend to other species for which chromosome maps and homologies have been worked out is discussed. PMID- 2980963 TI - Microdistribution of immature African blackflies resulting from water velocity and turbulence preferences. AB - 1. The velocity preferences of larval and pupal blackflies were studied experimentally by comparing the colonization of plastic strings placed in different velocity ranges; and also by investigating the simuliid microdistribution under natural conditions in the river. This required a novel method described for taking spot measurements of water velocity. 2. Simulium mcmahoni de Meillon, S. hirsutum Pomeroy and S. cervicornutum Pomeroy were most abundant in slow velocities (0.3-1.0 m s-1), S. colasbelcouri Grenier & Ovazza and S. hargreavesi Gibbins preferred 1.1-1.8 m s-1, S. squamosum Enderlein and S. vorax Pomeroy 1.9-2.2 m s-1. 3. The larvae of S. hargreavesi and S. cervicornutum preferred a higher velocity range compared with their pupae. 4. In moderate velocities (1.1-1.4 m s-1), the abundance of S. hargreavesi was greater under turbulent conditions than in smooth-flowing water, but this was reversed at very high velocities (2.3-2.6 m s-1). 5. Velocity had no apparent effect on substrate preference when substrates of different flexibilities were compared for three blackfly species. All avoided the most flexible substrates and preferred ones consisting of rigid articulating plates. Anisopteran predators were found to have a similar substrate choice. PMID- 2980964 TI - Phase shifting of the circadian conidiation rhythm in Neurospora crassa by calmodulin antagonists. AB - The effects of chemicals capable of antagonizing the functions of calmodulin, such as trifluoperazine, chlorpromazine, imipramine, alprenolol, W7, and W13, on the circadian conidiation rhythm of Neurospora crassa were examined. Trifluoperazine, at a 30-microM concentration, was most effective in shifting the phase of the conidiation rhythm and caused a maximum phase delay at circadian time (CT) 6 and maximum phase advance at CT 9. Chlorpromazine was less effective than trifluoperazine, and a 300-microM concentration of chlorpromazine was required for a similar phase shift. Imipramine, at a 1-mM concentration, caused only a small phase shift, while alprenolol had little effect on biological clock function. W7 and W13 caused phase delays longer than 10 hr at CT 6 and caused a phase advance of about 5 hr at CT 10 when present at a 200-microM concentration. However, W5 and W12, the dechlorinated homologues of W7 and W13, had no effects on clock function at the same concentration. Calmodulin was assayed by measurements of stimulation of cyclic nucleotide diphosphodiesterase activity. Calmodulin content remained constant in trifluoperazine-sensitive and trifluoperazine-insensitive phases for two cycles following the light-dark transition. PMID- 2980965 TI - Loss of temperature compensation of circadian period length in the frq-9 mutant of Neurospora crassa. AB - A new circadian clock mutant of Neurospora crassa has been isolated, whose most distinctive characteristic is the complete loss of temperature compensation of its period length. The Q10 of the period length was found to be equal to about 2 in the temperature range from 18 degrees to 30 degrees C. The period length was also found to be dependent on the composition of the medium, including the nature and concentration of both the carbon source and the nitrogen source. Although the rate of the clock and the growth rate were directly related when affected by varying the temperature, they were inversely related when altered by changing the composition of the medium. Therefore, the mutation has not simply coupled clock rate to growth rate in this strain. The mutation maps to the frq locus, where seven other clock mutations previously studied also map. Therefore, this mutant has been called frq-9. Since several of the other frq mutants show partial loss in temperature compensation, it is suggested that the frq gene or its product is closely related to the temperature compensation mechanism of the circadian clock of Neurospora. PMID- 2980966 TI - Expression of anti-DNA clonotypes and the role of helper T-lymphocytes during the autoimmune response in mice tolerant to alloantigens. AB - BALB/c mice neonatally injected with semiallogeneic (C57BL/6 x BALB/c) F1 spleen cells become tolerant to C57BL/6 alloantigens and exhibit chimaerism due to persistence of F1 lymphocytes. Such mouse chimaeras develop an autoimmune (lupus like) disease characterised by hypergammaglobulinaemia with production of autoantibodies against DNA, Sm antigen and other self-antigens characteristic of SLE in addition to circulating immune complexes and glomerular deposition of immunoglobulins. We have studied the autoimmune response by analysing the isoelectric focusing (IEF)+ patterns (spectrotypes) of anti-ss and anti-dsDNA antibodies produced by these animals. The results show that the anti-DNA response is remarkably restricted, only a very small number of lymphoid cell clones responding in the majority of animals. The behaviour of these clones has been followed during the development of the autoimmune response by analysis of their individual IEF patterns (clonotypes). The first appearance of clones secreting anti-DNA autoantibodies was observed in 3-4 week old mice. Changes in spectrotype occurred during the course of the response but they remained restricted to a very small number of clones in almost all the animals studied. Changes in clonotype consistent with somatic mutation in committed, anti-DNA-secreting clones were also observed. Helper T-lymphocytes of host origin are shown to be required for the development of an autoimmune response. PMID- 2980967 TI - Ranitidine and cimetidine in the healing of duodenal ulcer: meta-analysis of comparative clinical trials. AB - All available ranitidine and cimetidine comparative trials in acute duodenal ulcer disease were examined: of the 44 trials, 36 favoured ranitidine, and there was an overall difference in ulcer healing of 7%. Further stratification enabled examination of trials with common attributes: the most frequent endoscopic assessment was at 4 weeks to compare ranitidine 150 mg twice daily with cimetidine 1 g day-1 or 400 mg b.d. Twenty of these trials had sufficient data to permit pooling. Ranitidine was favoured in 18/20 trials and in three the differences achieved statistical significance. Results of the trials were combined using meta-analysis to calculate differences in ulcer healing. Most studies had sample sizes that were insufficient to detect clinically-important differences; the power to detect a 20% difference was less than 80% in 15/20 trials, and for a 10% difference was less than 80% in all but one trial. Fifteen trials compared ranitidine 150 mg b.d. with cimetidine 1 g day-1: healing after 4 weeks therapy was overall 6% greater for ranitidine. This was statistically significant (P less than 0.05) and the combined total number of patients had a power of 83% to detect this difference. In five trials the dose of cimetidine used was 400 mg b.d.: the 12% difference in healing in favour of ranitidine 150 mg b.d. was statistically significant, and the combined trials had a power of 95% to detect this difference. Ranitidine 150 mg twice daily heals significantly more duodenal ulcers after 4 weeks of therapy than either cimetidine 400 mg b.d. or cimetidine 1 g day-1. PMID- 2980968 TI - 5-Alkylresorcinols from Hakea amplexicaulis that cleave DNA. AB - A dichloromethane extract of Hakea amplexicaulis was found to cause strand scission of phi X174 replicative form DNA in the presence of Cu(II). Bioassay guided fractionation of this extract afforded five compounds capable of mediating DNA relaxation. Structure determination of the active principles indicated that they were 5-tridecylresorcinol (1), 5-pentadec-cis-8-enylresorcinol (2), 5 heptadeca-8,11-dienylresorcinol (3), 5-pentadecylresorcinol (4), and 5-heptadec cis-8-enylresorcinol (5). As noted previously for compounds in this structural series, DNA cleavage was enhanced significantly by incubation under (alkaline) conditions known to promote oxygenation of the parent compounds on the aromatic nucleus. PMID- 2980969 TI - The Konno procedure for enlargement of the aortic root. PMID- 2980970 TI - Surgical treatment of double inlet ventricle ("single ventricle"). AB - The surgical techniques described are the result of an evolution over a number of years in the performance of the septation operation and the modified Fontan Kreutzer repair for patients with double inlet ventricles. Those with associated pulmonary stenosis are best palliated by a classical Blalock-Taussig or Goretex shunt if an operation is required during the first few years of life and later, between two and four years of age, definitive repair by the modified Fontan Kreutzer operation is advised. Although controversial, we prefer the use of a large nonvalved right atrial-pulmonary artery connection. Ventricular septation remains the best definitive surgical option when pulmonary stenosis is absent or mild. It is contraindicated by severe pulmonary vascular disease and also by less than moderate ventricular enlargement. The need for concomitant AV valve replacement and the use of an extracardiac conduit are associated with increased hospital mortality in our experience. Infants identified during the first year of life who do not have pulmonary stenosis are a difficult subset to manage. If the VSD and subaortic area is large and unobstructed, pulmonary artery banding early in life will control pulmonary vascular resistance and from this standpoint, permit these patients to become ultimately suited to a modified Fontan-Kreutzer repair. Unfortunately, ventricular hypertrophy usually results from pulmonary artery banding and has been associated with higher hospital mortality at the time of definitive repair. When pulmonary artery banding is undertaken for this subset, debanding and definitive repair seems best advised at about two years of age. Pulmonary artery banding is well known to accelerate the development of subaortic stenosis by spontaneous progressive restriction of the VSD. This results in small ventricular cavity size and increased ventricular hypertrophy, which are incremental risk factors for increased hospital mortality by either definitive procedure. When the VSD or subaortic area is narrow and the patient is identified during the first year of life, isolated pulmonary artery banding is inappropriate. The surgical options for these patients include Ebert's two-stage management program consisting of the initial placement of a loose partial septation patch with concomitant pulmonary artery banding, and later debanding and complete septation. Alternatively, a trial of primary complete septation may be warranted, or the use of a procedure consisting of division of the main pulmonary artery with distal closure and anastomosis of the proximal portion to the side of the ascending aorta, coupled with a systemic-pulmonary artery shunt. PMID- 2980972 TI - Tricuspid valve repair using a flexible linear reducer. PMID- 2980971 TI - Closure of ventricular septal defect in infancy. PMID- 2980973 TI - Cryopreserved pulmonary homograft valved external conduits: early results. AB - Cryopreserved homograft pulmonary valves were used as a valved external conduit between the right ventricle and native pulmonary arteries in 21 patients with complex congenital heart disease. The technique used to construct the conduit was modified after incompetence of the valve was detected early in 3 of the first 9 conduits. The next 12 conduits remained competent. This experience suggests that early valve competency can be achieved if the valve is not distorted during construction of the conduit. PMID- 2980974 TI - Unsatisfactory clinical experience with a collagen-sealed knitted Dacron extracardiac conduit. AB - A clinical trial of a collagen-sealed knitted Dacron conduit (Tascon Medical Technologies) in 86 patients has revealed a high incidence of early reoperation for conduit stenosis. At 3 years, the actuarial incidence of freedom from conduit replacement was 67 +/- 14% for valved conduits, and 66 +/- 20% for nonvalved conduits. Seven of eight conduits that were replaced had a thick, weakly adherent pseudointima. Comparison of the current series with a previous series of patients receiving tightly woven low-porosity Dacron conduits is complicated by the young age, small size, and greater complexity of the current group. Nevertheless, the findings are consistent with the results of two laboratory studies performed at this hospital which suggest that the collagen used in the Tascon conduit undergoes particularly slow resorption, resulting in weak adhesion between the pseudointima and conduit. This allows hemorrhagic dissection to occur deep to the pseudointima. These data suggest that alternative methods of sealing knitted Dacron conduits should be used. PMID- 2980975 TI - The physiological basis of left ventricular diastolic dysfunction. AB - Overall cardiac pump function requires adequate ventricular diastolic filling as well as normal systolic ejection. Abnormalities of the rate or extent of myocardial relaxation (diastolic dysfunction) have been described in a large variety of clinical conditions, including hypertrophy, ischemia, and after cardiac surgery. Diastolic and systolic dysfunction can be readily distinguished by analysis of pressure volume loops and utilization of echocardiography or nuclear cardiology gated blood pool scans. The mechanisms by which diastolic dysfunction can occur may be structural (hypertrophy, fibrosis) or dynamic (hypoxia, ischemia, alteration of diastolic cytosolic calcium levels). Hypertrophied myocardium is particularly susceptible to diastolic dysfunction by virtue of both structural changes (increased LV mass and interstitial fibrosis) and greater susceptibility to develop impaired myocardial relaxation during hypoxia or ischemia than nonhypertrophied myocardium. PMID- 2980976 TI - Low back pain: development and five-year prospective application of a computerized quality-based diagnostic and treatment protocol. AB - A standardized protocol for low back pain was developed and computerized so that it could be used as a concurrent monitoring system for large patient populations. The software incorporated a relational database management system (RDBMS) and C language, a flexible, general-purpose programming language that is fast and portable. The protocol was then applied to a uniform group of industrial patients for 5 years. The results demonstrated that a quality-based protocol could be successfully computerized and applied to a large group of patients as a concurrent monitoring system. Quality care was insured by adherence to the computerized protocol. Associated economic results showed decreases in actual number of accidents each year (from 98/year prior to the study to 42/year in the last study year), in lost work days (from 3640/year before the study to 2118/year in the last year), and in costs (savings averaged $430,000/year). The goal of this study was to provide quality medicine; the economic benefits were a bonus. The monitoring system differed from those of the past in that it was driven by the basic medical information taken from the history, physical examination, and radiograph findings. The monitoring physicians were unbiased because they were not allowed to become involved in the patient's ongoing care. There was no rebound phenomenon (an initial drop in cases followed by a gradual return to the prestudy level); this was attributed to the constant monitoring of each case from beginning to end. These results led to the following conclusions: (a) Computerization of a standardized medical approach for low back pain is practical.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2980977 TI - Population relationships of Simulium yahense and Simulium sanctipauli in the Firestone Rubber Plantation at Harbel, Liberia. AB - Comparative genetic differences for the phosphoglucomutase and trehalase loci were surveyed in larval and adult blackfly populations of the onchocerciasis vectors Simulium yahense Vajime and Dunbar, and S.sanctipauli Vajime and Dunbar. Genotype frequencies for each stage and all populations were at Hardy-Weinberg equilibrium, indicating that S.yahense and S.sanctipauli remain genetically distinct. S.yahense populations from three different locations were found to be genotypically comparable. The larval population of S.yahense found in closest proximity to a S.sanctipauli population was found to express the phosphoglucomutase allele 1.33, characteristic of S.sanctipauli, with significantly greater frequency than other larval populations of S.yahense. This may constitute evidence of limited genetic introgression of S.sanctipauli with the S.yahense population. PMID- 2980978 TI - Solitary spinal plasmacytomas: management and outcome. AB - Nine patients with solitary plasmacytoma of the spine were reviewed. Four of these patients progressed to multiple myeloma within 9 +/- 4 months (mean +/- SD) from diagnosis, and died from their disease in 23 +/- 15 months. In contrast to this, the five remaining patients free of systemic disease or local recurrence survived 78 +/- 66 months. No correlation was found between age at diagnosis, lesion location, symptomatology, laboratory studies, surgical treatment, or radiation dosage and progression to systemic disease. Solitary plasmacytoma will progress to systemic multiple myeloma and death in approximately 50% of patients. The remaining patients exhibit long-term survival following local treatment of the solitary lesion. PMID- 2980979 TI - Complications in the first 40 cases of microdiscectomy. AB - From 1984 to 1986, a single surgeon treated 40 patients suffering from lumbar disc herniation by the technique of microdiscectomy. While the advantages of working under magnification are endorsed, the complications encountered are analysed with the aim of giving warning to surgeons keen to start the technique. The complications included intraoperative dural tears (two cases), discitis (two cases), and recurrence of back pain (two cases). The intraoperative dural damage did not require specific treatment, while the patients with discitis responded readily with antibiotics. The recurrent cases were found to be caused by adhesion bands produced by hanging tags of incompletely removed yellow ligament. It is therefore concluded that in microdiscectomy prophylactic antibiotics should be given and a complete removal of the yellow ligament on the side of surgery should be ensured to avoid infection and recurrent root impingement. PMID- 2980980 TI - Membrane properties of bovine airway smooth muscle cells: effects of maturation. AB - Airway smooth muscle cells of bovine trachealis muscle from immature (2 weeks old), developing (5 months old) and mature animals (greater than 5 years old) have resting membrane potentials of -63.5 +/- 0.4 mV, -62.2 +/- 0.8 mV and -60.3 +/- 0.7 mV, respectively. These resting potentials were not significantly different. The contribution of the electrogenic sodium pump to the resting membrane potential of airway smooth muscle cells decreases significantly with age being 38.3% +/- 0.9, 30.6% +/- 0.8 and 21.3% +/- 0.9 in tissues from immature, developing and mature cows, respectively. The contribution of the electrogenic sodium pump to resting membrane potential was not influenced by the degree of stretch (applied load) of the airway smooth muscle cells. However, resting membrane potential was reduced with increasing tone especially when loads between 5 to 10 g were applied. The densities of Na+/K+ pump sites, as measured by [3H] ouabain binding, decreased significantly as a consequence of maturation and were 4.12 +/- 5.23, 3.08 +/- 0.26 and 1.94 +/- 0.38 pmoles/mg protein ouabain binding in tissues from immature, developing and mature animals respectively. The affinity of the pump sites for the cardiac glycoside, ouabain, did not change during maturation. We conclude that maturation alters both the number of Na+/K+ pump sites and the contribution of the electrogenic sodium pump to the membrane potential of airway smooth muscle cells. These age related changes may contribute to the reduced airway reactivity to both bronchoconstrictor and bronchodilator agents previously observed both in vitro and in vivo. PMID- 2980981 TI - Bronchoprotective and bronchodilator activity of anaritide (human atrial natriuretic factor [102-126]) infusion in the anesthetized guinea pig. AB - The present study examined the bronchoprotective and bronchodilator efficacy of infused human atrial natriuretic factor [102-126] (Anaritide) in the guinea pig. Anesthetized and paralyzed male guinea pigs, ventilated via a tracheal cannula, were placed in a plethysmograph for measurement of agonist-induced changes in pulmonary mechanics. Reductions in dynamic compliance of the lung and airway conductance were produced either by 3 to 7 tidal volume breaths of leukotriene D4 (0.125 micrograms/ml) delivered through an ultrasonic nebulizer or by intravenous histamine (2.8 +/- 0.2 micrograms/kg). Infusion of Anaritide (1 microgram/kg/min), before evoking bronchoconstriction with either LTD4 or histamine, produced a significant protection against bronchoconstriction produced by aerosol LTD4, but not against histamine-induced bronchoconstriction of similar magnitude. In other experiments, Anaritide infusion (0.5-2 micrograms/kg/min) also rapidly reversed a stable LTD4-induced decrease in airway conductance, but did not produce a similar reversal of the decrease in dynamic compliance. The data provide evidence that intravenous Anaritide exerts both bronchoprotective and bronchodilator effects against LTD4-induced bronchospasm. PMID- 2980987 TI - Financial implications of half- and full-time employment for persons with disabilities. A response to Schloss, Wolf and Schloss. PMID- 2980988 TI - Transplacental pharmacokinetics of teratogenic doses of etretinate and other aromatic retinoids in mice. AB - The transplacental pharmacokinetics of single teratogenic doses of etretinate and motretinide were compared with particular emphasis on distribution and concentrations in the exposed embryos of the free acid metabolite, etretin. The three aromatic retinoids were also tested for their direct inhibitory effect on chondrogenesis in the limb bud mesenchymal cell "micromass" culture assay. After a standard dose of 100 mg/kg administered on day 11 of gestation in NMRI mice, all three compounds were teratogenic, but they differed from each other in potency. Etretinate was most active as a teratogen, equalling the potency of our standard all-trans-retinoic acid; every exposed fetus was deformed with severe shortening of all limb bones as well as cleft palate. Etretin was less potent than etretinate, and motretinide was considerably less active as a teratogen than the other two. In the in vitro assay, only etretin suppressed chondrogenesis and this activity was equivalent to that of all-trans-retinoic acid (IC50 of 12 ng/ml). Both etretinate and motretinide (which contain an ethyl ester and ethylamide terminal group, respectively) were essentially inactive in vitro, demonstrating the fact that a free carboxylic group may be a requirement for the in vitro suppression of chondrogenesis. These differences between the results obtained in vivo and in vitro could be resolved by pharmacokinetic investigations using HPLC methods. Both etretinate and motretinide were metabolized in vivo to etretin, their likely common teratogenic metabolite. The high teratogenic potency of etretinate was probably the result of high concentrations as well as AUC values of its metabolite etretin in the embryo. On the other hand, the comparatively low teratogenicity of motretinide could be related to approximately 5 x lower embryonic peak levels as well as AUC values of etretin. A comparison of these results with those previously obtained for all-trans- and 13-cis-retinoic acids confirms the correlation between embryonic exposure and teratogenic potency in the mouse. Our results indicate that pharmacokinetic studies are essential for the interpretation of relative teratogenic potencies of retinoids as well as apparent differences between in vivo and in vitro teratogenesis. A free carboxyl group at the terminal end of the tetraene chain was necessary for high activity of the retinoids studied. PMID- 2980989 TI - Percutaneous transluminal angioplasty of the subclavian and vertebral arteries. AB - Percutaneous transluminal angioplasty (PTA) for atherosclerotic lesions of the subclavian, vertebral, and brachiocephalic vessels is being performed in selected cases. Clinically patients presented with symptoms of vertebral basilar insufficiency, multiple transient ischemic attacks, subclavian steal syndrome, and motor weakness of the upper extremity. Thus far, 22 procedures have been performed with good results. This included 16 subclavian, 4 vertebral, and 2 innominate arteries. Follow-up has ranged from 8 to 26 months (mean 17 months). No permanent neurologic complications have occurred with this technique, and all patients had significant clinical and radiographic improvement following this procedure. PMID- 2980990 TI - Practical aspects of percutaneous transluminal angioplasty of the carotid artery. AB - The authors are reporting a total of 41 patients undergoing percutaneous transluminal angioplasty (PTA) at different levels of the carotid artery. The procedure for proximal carotid artery was performed on 17 patients, for mid or distal common carotid artery on 4 patients, for carotid bifurcations on 5 patients, for internal carotid artery on 6 patients, for external carotid artery on 4 patients, and for fibromuscular dysplasia of the internal carotid artery on 5 patients. We also collected research on an additional 53 patients from the literature for a total of 94 patients. To date, we have not encountered any mortalities nor neurologic complications in performing this procedure. Thus, we believe that PTA may be performed with reasonable safety and as an alternate procedure in the treatment of carotid artery stenosis. PMID- 2980991 TI - A methodologic assessment of studies comparing magnetic resonance imaging and computed tomography of the brain. AB - To evaluate the methods used in studies comparing magnetic resonance imaging (MRI) and computed tomography (CT) of the brain, we reviewed English language articles published between 1981 and early 1986. Of 83 studies comparing the accuracy of MRI and CT in 10 or more patients, few met methodologic standards for evaluations of the efficacy of new diagnostic tests. Limitations were found in how study populations were assembled, in test performance and interpretation, and in analysis of study findings. Greater attention to design and execution of comparison studies could overcome these limitations and help demonstrate MRI's true diagnostic value. PMID- 2980993 TI - Volume flow measurement of pulsatile flow by digitised cine angiography. AB - The validation, by phantom experiments, of a method of measuring volumetric, pulsatile blood flow in vessels of about 4 mm calibre is described. 'Instantaneous' as well as integrated flow is obtained from cine angiograms by digitisation of the images and densitometric analysis in an advanced computer vision analysis system. From 48 frames per second cine, pulsatile flow at any moment between 0 and about 400 ml/min was calculated within about 10 per cent of the electromagnetic flow meter reading. Further improvement is expected. It is proposed to advance to 3D and have the methodology available for clinical use in due course. PMID- 2980992 TI - Fluorocarbon emulsion as a potential contrast medium in the subarachnoid space and brain tissue. Experiments in cats. AB - Ventriculosubarachnoid perfusion with oxygenated fluorocarbon emulsion has been used in the treatment of severe cerebral ischemia in cats. The perfusion vehicle is prepared mimicking cerebrospinal fluid in terms of electrolyte content, osmolality, pH, glucose and amino acids. Fluorocarbon emulsion is prepared with the vehicle as the external phase and charged with high partial pressures of oxygen. This fluorocarbon emulsion has been well-tolerated by the central nervous system. In addition to delivering oxygen to the ischemic tissues via the ventriculosubarachnoid space, the fluorocarbon emulsion has an absorption coefficient higher than normal brain and cerebrospinal fluid. This property makes it easily detected on the CT scan. Determination of appearance, attenuation, distribution, and absorption of the fluorocarbon has been performed in cats after direct intraventricular injection of the substances and after perfusion of the emulsion. The results of these experiments stress the possibility of using a fluorocarbon emulsion as a potential contrast medium in the brain and subarachnoid space, particularly after brain anoxia. PMID- 2980994 TI - Recurrent post-discectomy pain. CT--surgical correlation. AB - Diagnostic evaluation of the patient with recurrent lumbosacral radiculopathy following lumbar laminectomy and discectomy represents a difficult task for the radiologist. Neither non-enhanced CT nor myelography have permitted reliable differentiation between postoperative extradural fibrosis (scarring) and recurrent disc herniation. However, CT following intravenous contrast infusion has been reported by several investigators to allow differentiation of these two entities. A series of 13 patients is reported with correlation between the findings on postcontrast CT and at reoperation. In 5 patients who exhibited variable contrast enhancement of an extradural soft tissue mass without a discreet lucent component on postcontrast CT, operative reexploration revealed only scar tissue. In 8 patients in whom postcontrast CT demonstrated a discreet non-enhancing extradural mass (partially surrounded by a thin enhancing rim of soft tissue in all but one), recurrent disc herniation together with an overlying band of scar was found and resected at reoperation. The strong positive correlation between CT and operative findings in this series suggests that postcontrast CT may well prove to be a reliable method for preoperative differentiation of recurrent disc herniation from extradural scarring. PMID- 2980996 TI - Dental volunteers for Israel: a children's free dental clinic in Jerusalem. PMID- 2980995 TI - [Progress in endocrinology]. PMID- 2980997 TI - Poison ivy dermatitis in Arizona. PMID- 2980998 TI - Biopsy evaluation of chronic active hepatitis. Peritoneoscopy with directed liver biopsy versus blind percutaneous liver biopsy. AB - In this prospective study of chronic active liver disease, we compared the assessment of hepatic histology in samples obtained by peritoneoscopy with directed liver biopsy and blind percutaneous liver biopsy in 23 cases (22 patients, one patient studied twice). In blinded fashion, a pathologist assessed all specimens for evidence of cirrhosis and degree of necroinflammatory change. Two clinicians independently reviewed clinical and laboratory findings in both sets of biopsies. Each committed in writing recommendations regarding immunosuppressive therapy, follow-up interval, and rebiopsy date. The final diagnosis differed from that made by percutaneous and directed biopsy in 2 of 23 (9%) and 1 of 23 (4%) cases, respectively. Six cases of cirrhosis were correctly diagnosed by both biopsy methods, but only four of the six cirrhotic cases were diagnosed by gross peritoneoscopic findings. In only 2 of 23 (9%) cases was there disagreement in the degree of necroinflammatory change between the blind and directed biopsies that affected treatment recommendations. We conclude that blind percutaneous biopsy adequately diagnoses and monitors activity in viral chronic hepatitis for treatment purposes. PMID- 2980999 TI - Compliance among heavy alcohol users in clinical drug trials. AB - A high level of patient compliance is essential in determining the efficacy and safety of treatment with new drugs. Alcohol abusers are generally considered to exhibit a low level of compliance with respect to the taking of medication. This study describes a selection strategy and compares methods of determining compliance in two clinical trials assessing the effects of zimelidine and citalopram on ethanol intake in nondepressed male heavy drinkers (greater than 4 drinks/day). Objective methods included the use of a tracer substance (riboflavin), measurement of neurochemical effects (serotonin uptake inhibition), and measurement of drug and metabolite concentrations in body fluids (blood and urine). Other methods included self-report and pill count. Mean compliance estimates for the taking of medication ranged from 78 to 97% depending on the method used. Methods to determine ethanol use included daily self-monitoring and daily urine alcohol measurement. Alcohol consumption correlated with urine alcohol measurement (r = 0.62, p less than .001). These comprehensive assessments of compliance allowed accurate evaluations of drug efficacy and toxicity. The study demonstrates that with stringent selection and reinforcement procedures, exceptionally high levels of compliance can be attained in this group of subjects. Similar procedures should be included in trials evaluating new drugs. PMID- 2981000 TI - [A method of long-term preservation of erythrocytes by freezing at moderately low (-60, -40 degrees C) temperatures using a Russian-made low-temperature cooler KLGKh]. PMID- 2981001 TI - [Effect of NaCl on the survival of Campylobacter jejuni at a temperature of 20 degrees Centigrade]. PMID- 2981002 TI - Endocarditis due to six entrapped pacemaker leads and concomitant recurrent coronary arteriosclerosis. AB - A case of endocarditis associated with six entrapped endocardial pacer leads is presented. Because of many failed attempts at removing them by conservative measures, cardiopulmonary bypass was needed; concomitant redo coronary bypass grafts were done. To our knowledge, this represents a unique case, the like of which has not been reported previously. Salient features of management are discussed. PMID- 2981003 TI - [Determination of immunocomplexes. IV. Detection of immune complexes using the C3 and C4 component of complement]. PMID- 2981004 TI - [Use of new rapid examination methods in the diagnosis and therapy of purulent meningitis]. PMID- 2981007 TI - Optimal treatment of prostate cancer. PMID- 2981006 TI - Let's test the validity of second-look laparotomy. PMID- 2981005 TI - [Effect on Haemophilus influenzae of its transforming deoxyribonucleic acid irradiated in vitro using ultraviolet light at -70 degrees]. PMID- 2981008 TI - [Surgical treatment of 180 degrees giant tears. Apropos of 5 cases]. PMID- 2981009 TI - Current opinion in contemporary orthodontics. PMID- 2981011 TI - [The prevention of dry socket]. PMID- 2981010 TI - [Effect of mechanical deboning process on the fat stability of 3 tropical fish species stored at -10 degrees C]. AB - Maximum advantage of certain fish species captured with shrimp, at present not fully utilized, constitutes a necessity in Venezuela. The purpose of this research work was to evaluate the stability of three of those fish species: bagre, cunaro and caballa, based on the changes that might occur in their fat content. This was done by comparing both the edible part of the whole fish, as well as the deboned flesh of each of them. Evaluation of the changes was performed by means of the acid and peroxide indices, with previous fat extraction, and the thiobarbituric acid (TBA) index. On the other hand, the fatty acids profile of the three species studied was determined by gas chromatography. According to our findings, the greater alteration was detected in the deboned sample, obtaining the highest TBA and acidity index values of all species, on the third month of storage; then a reduction of these values was observed. On the other hand, the peroxide index presented heterogeneous values in the three species analyzed through their storage period. The predominant fatty acids were: palmitic (16:0) among the saturated, and oleic (18:1) among the unsaturated. A greater proportion of unsaturated fatty acids was found in the three species. An increment of these in the deboned sample with respect to the whole fish, was also observed. PMID- 2981012 TI - External craniofacial fixation treatment for mid-facial fractures? Part I: Classification of mid-facial fractures, types of occlusion and treatment methods. PMID- 2981013 TI - An acid-etch fixed prosthesis. PMID- 2981014 TI - A new retrobulbar needle and injection technique. AB - A new needle and method have been developed for the administration of retrobulbar injections. The needle is curved as it extends from its hub, but it has a straight terminal portion. Use of this needle with the technique described should decrease the risk of most complications associated with conventional needles and techniques. PMID- 2981015 TI - Basal cell carcinoma with follicular differentiation. AB - A distinctive form of basal cell carcinoma with follicular differentiation was studied in 15 biopsy specimens. The neoplasms differ from other variants of basal cell carcinoma by being situated on the face only, being small and well circumscribed, and having signs of follicular differentiation, namely, numerous tiny infundibular cyst-like structures and aggregations of neoplastic cells that resemble hair follicles in telogen. The diagnostic features of basal cell carcinomas with follicular differentiation are enumerated and illustrated. PMID- 2981016 TI - Heat and moisture exchanging bacterial filters. PMID- 2981018 TI - Basal cell carcinoma with follicular differentiation. PMID- 2981017 TI - Assessing quality. PMID- 2981019 TI - Dental chairside assistants: a report on a survey. PMID- 2981020 TI - Preventive dental health behaviour of mothers as role models for their children. PMID- 2981021 TI - Conservative management of an odontogenic keratocyst: a four and a half year evaluation. PMID- 2981022 TI - General practitioner's radiology casebook XI. Lymph node calcification. PMID- 2981023 TI - Immunological abnormalities in human interleukin-2 or interleukin-2/interleukin-2 receptor L chain transgenic mice. AB - We constructed two strains of transgenic mice that carry the cDNA of either human interleukin-2 (IL-2) or the L chain of its receptor under the control of the H 2Kd promoter. The IL-2 transgenic mice expressed human IL-2 mRNA in the thymus, spleen, bone marrow, lung, muscle, and skin. Human IL-2 protein was also detected in their sera. The IL-2 transgenic mice suffered from dermatitis and pneumonia. Immune responses of their spleen cells against antigens were significantly impaired whereas their spleen cells responded well to polyclonal lymphocyte activators, suggesting that constitutive expression of IL-2 might have affected the repertoire formation of T cells in the mice. The IL-2 receptor (IL-2R) transgenic mice were healthy. We crossed the IL-2 and IL-2R transgenic mice to yield hybrid mice expressing both the ligand and the receptor constitutively. The life span of the hybrid mice was remarkably shortened. In addition to several abnormalities found in the IL-2 transgenic mice, spleen cells of the hybrid mice showed the strong natural killer activity which was ascribed to a large number (18%) of Thy-1+/CD3-cells unique to their spleen. PMID- 2981024 TI - [Diagnostic worth of panoramic X-rays for the edentulous patient]. PMID- 2981025 TI - Professor GHR von Koenigswald: his contribution to knowledge of the evolution of man and his dentition. PMID- 2981027 TI - The influence of dental practice characteristics on pit and fissure sealant use: a national survey. PMID- 2981026 TI - Influence of the "squeeze jacket" on lung function in young infants. AB - The possible influence of the "squeeze jacket" on measurements of lung function in young infants was examined in 29 infants. In 19 infants, a comparison was made between jacket off and jacket on with the arms held inside the jacket. With the jacket on (arms inside), we found a significant reduction in thoracic gas volume (TGV). Passive compliance of the respiratory system (Crs) was also significantly decreased, while the resistance of the respiratory system (Rrs) was significantly increased. An additional 10 infants were studied comparing jacket off with jacket on (arms outside) and jacket on (arms inside). TGV was significantly less with the jacket on (arms inside) compared to jacket off and jacket on (arms outside), while Rrs and mouth pressure at end-inspiratory occlusion (Pm) were significantly increased. There was no difference between jacket off and jacket on (arms outside). These observations suggest that the squeeze jacket with the arms held at the infant's side inside the jacket exerts a restricting effect on chest and abdominal wall movement and leads to errors in the measurements. To avoid these potential errors, measurements should be made without the jacket on or with the arms held outside the jacket. PMID- 2981029 TI - SPECT image noise power: effects of nonstationary projection noise and attenuation compensation. AB - The effects of nonstationary projection noise and attenuation compensation are included in a theoretic calculation of the radial noise power spectrum (NPS) of single photon emission computed tomographic (SPECT) images. The nonstationary projection noise is shown to cause a relatively large d.c. component in the NPS, especially for small objects; whereas, attenuation compensation increases the total noise variance while only changing the d.c. component slightly. The theoretic calculation agrees well with a radial NPS estimated from 1,250 SPECT images simulated from projections of random Gaussian noise, even though the effects of discrete data collection and reconstruction were not included in the theoretical model. PMID- 2981028 TI - [Modification of the process of resorption of alpha-emitting nuclides from the gastrointestinal tract depending upon the properties of administered compounds and the physiological state of rats]. AB - In experiments on rats a study was made of resorption from the gastrointestinal tract of simple salts, complex compounds and "biologically incorporated" forms of transuranium nuclides, the influence of age, pregnancy and chemically active substances (for instance, trivalent iron and ethyl alcohol) on this process. PMID- 2981030 TI - Stress and the student. PMID- 2981031 TI - Osteomalacia and coeliac disease presenting as isolated dactylitis. AB - A 30 year old Pakistani female patient with osteomalacia and coeliac disease presenting as an isolated dactylitis is reported. PMID- 2981032 TI - Childhood illness as an aetiological factor in burns. AB - A prospective study of 208 children has shown that a significant number of them have an illness before suffering a thermal injury. The authors suggest that an unwell child is more likely to suffer a burn, and that first aid programmes should stress to parents to be even more careful when their child is unwell. PMID- 2981033 TI - [Bluish pigmentation of the nails in AIDS patients. Possibly an adverse effect of Zidovudine?]. PMID- 2981034 TI - [Levels of ALAT and ASAT in a population of altruistic blood donors]. PMID- 2981035 TI - [Percutaneous endoscopic gastrostomy. Indications and follow-up in 52 patients]. AB - A nutritional support was indicated in 52 patients, most of whom had deglutition problems linked with post-therapeutic after-effects of neoplasia of the ORL sphere. Assistance was possible in all the cases, with difficulty in 9 cases. There were only three major complication (two of which necessitated the removal of the tube) and no fatalities. Benign, more numerous complications were all medically curable and did not necessitate the removal of the tube. The follow-up of the patients showed remarkable tolerance to, and nonproblematic use of, the GPE tube as a nutritional aid leading, in the majority of cases, to a weight gain, even when ambulatory. Our experience suggests that this technique deserves a prime place as a nutritional aid for cases of deglutition problems, of neurological origin, and in ORL cancers. Its relative safety must undoubtedly make it preferable to the surgical technique. PMID- 2981036 TI - [The temporal arteritis syndrome]. PMID- 2981037 TI - Assessment of hand trauma. PMID- 2981038 TI - Multiple trauma: a case study using an air fluidized support system. PMID- 2981039 TI - Management of the patient with renal osteodystrophy. PMID- 2981040 TI - Complications of Metrizamide (amipaque) myelography. PMID- 2981041 TI - The adolescent with osteogenic sarcoma. PMID- 2981042 TI - Adolescent adjustment to amputation necessitated by bone cancers. PMID- 2981043 TI - Total contact orthosis for immobilization of thoracolumbar fractures. PMID- 2981044 TI - Do you practice what you (preach) write? Legal questions and answers. PMID- 2981045 TI - Coherence treatment of postmenopausal osteoporosis with growth hormone and calcitonin. AB - Fourteen women with postmenopausal osteoporosis, all having at least one vertebral crush fracture, were randomly assigned to two treatment arms, each lasting 24 months. The coherence treatment group (7 patients) was treated in the following sequence: human growth hormone (hGH) 7 IU subcutaneously daily for 2 months, followed by 3 months of salmon calcitonin (CT), 100 MRC units every other day. After a 3 month rest period, this sequence was repeated twice. The contrast group (7 patients) was treated intermittently with salmon CT given in the same time periods and at the same dose as in the coherence treatment group. Bone mass was measured every 4 months by neutron activation analysis for total body calcium (TBCa) and by single photon absorptiometry for bone mineral content (BMC) of the distal radius. Although there were no significant differences between the two groups (two-way ANOVA), the rate of change in TBCa in the coherence treatment group was significantly different from zero (F = 3.8, P less than .05) and was +2.3%/year. The increase in bone mass appeared to be sustained throughout the 2 year study, in contrast with previous studies where a plateau effect was observed with calcitonin given alone or continuously with growth hormone. No significant change was found in bone histomorphometric values measured before and after treatment in 4 patients from each group. PMID- 2981046 TI - A plasmid of Rhizobium meliloti 41 encodes catabolism of two compounds from root exudate of Calystegium sepium. AB - Our objectives were to identify substances produced by plant roots that might act as nutritional mediators of specific plant-bacterium relationships and to delineate the bacterial genes responsible for catabolizing these substances. We discovered new compounds, which we call calystegins, that have the characteristics of nutritional mediators. They were detected in only 3 of 105 species of higher plants examined: Calystegia sepium, Convolvulus arvensis (both of the Convolvulaceae family), and Atropa belladonna. Calystegins are abundant in organs in contact with the rhizosphere and are not found, or are observed only in small quantities, in aerial plant parts. Just as the synthesis of calystegins is infrequent in the plant kingdom, their catabolism is rare among rhizosphere bacteria that associate with plants and influence their growth. Of 42 such bacteria tested, only one (Rhizobium meliloti 41) was able to catabolize calystegins and use them as a sole source of carbon and nitrogen. The calystegin catabolism gene(s) (cac) in this strain is located on a self-transmissible plasmid (pRme41a), which is not essential to nitrogen-fixing symbiosis with legumes. We suggest that under natural conditions calystegins provide an exclusive carbon and nitrogen source to rhizosphere bacteria which are able to catabolize these compounds. Calystegins (and the corresponding microbial catabolic genes) might be used to analyze and possibly modify rhizosphere ecology. PMID- 2981047 TI - High expression of an N-acetylglucosaminyltransferase III in 3'-methyl DAB induced hepatoma and ascites hepatoma. AB - Ascites hepatoma AH-66 and 3'-Me-DAB-induced hepatoma of rats contain highly active N-acetylglucosaminyltransferase III (GnT-III) which catalyzes the addition of N-acetylglucosamine through a beta 1-4 linkage (bisecting N-acetylglucosamine) to the beta-linked mannose of the trimannosyl core of asparagine-linked sugar chains, whereas normal rat liver contains very little. The high activity was also detected in fetal rat liver, newborn rat liver, hyperplastic nodules and various transplantable hepatomas. PMID- 2981049 TI - Developmental trends in vocal fundamental frequency of young children. AB - Fundamental frequency (F0) values are reported for 14 children between the ages of 11 and 25 months, an age period characterized by changes in physiological and linguistic development. Subjects were grouped into 3-month age intervals reflecting a continuum of physical development and were audiotape-recorded during spontaneous speech productions. Acoustic analysis of average F0 and F0 variability was performed. F0 variability was found to decrease as subject age increased, as did segment durations. The decrease in average F0 across the arbitrary age groups did not reach significance; however, when viewed within the overall developmental period and in comparison with data from other studies of younger and older children, average F0 during this age is consistent with a decreasing trend throughout early childhood. PMID- 2981050 TI - Ca2+-mediated generation of inositol 1,4,5-triphosphate and inositol 1,3,4,5 tetrakisphosphate in pancreatic islets. Studies with K+, glucose, and carbamylcholine. AB - The role of Ca2+ in the generation of inositol phosphates was investigated using rat pancreatic islets after steady state labeling with myo-[2-3H]inositol. Depolarizing K+ concentrations (24 mM) evoked early (2 s) increases in inositol 1,4,5-trisphosphate (Ins-1,4,5-P3) and inositol 1,3,4,5-tetrakisphosphate (Ins 1,3,4,5-P4) as measured by high performance anion-exchange chromatography. The increase in Ins-1,4,5-P3 was transient and was followed by a more pronounced rise in Ins-1,3,4-P3. These effects were dependent on the presence of extracellular Ca2+ but were not secondary to release of either neurotransmitters or metabolites of arachidonic acid. K+ also promoted the breakdown of phosphatidylinositol 4,5 bisphosphate (PtdIns-4,5-P2) and of the other phosphoinositides. Glucose (16.7 mM) was less marked in its effects but still promoted rapid increases in Ins 1,3,4,5-P4 (2 s) and Ins-1,4,5-P3 (10 s) and a slower rise in Ins-1,3,4-P3 (30 s). The levels of all three metabolites rose steadily over 10 min stimulation. These responses to glucose could be largely, although not entirely, inhibited by depletion of extracellular Ca2+ or by Ca2+ channel blockade with verapamil (20 microM). Carbamylcholine (0.5 mM) was the most potent stimulus used evoking early rises in Ins-1,4,5-P3 and Ins-1,3,4,5-P4 (2 s) followed by Ins-1,3,4-P3 (10 s), effects which were only partially dependent on extracellular Ca2+. The results suggest that a Ca2+-mediated PtdIns-4,5-P2 hydrolysis accounts for most of the Ins-1,4,5-P3 generated in response to glucose but not carbamylcholine. In addition, glucose may exert effects on inositol phosphate metabolism which are Ca2+ independent. PMID- 2981051 TI - Inositol trisphosphate isomers, but not inositol 1,3,4,5-tetrakisphosphate, induce calcium influx in Xenopus laevis oocytes. AB - To investigate the mechanisms by which inositol phosphates regulate cytosolic free Ca2+ concentration ([Ca2+]c), we injected Xenopus oocytes with inositol phosphates and measured Ca2+-activated Cl- currents as an assay of [Ca2+]c. Inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) injection (0.1-10.0 pmol) induced an initial transient Cl- current (I1) followed by a second more prolonged Cl- current (I2). Both currents were Ca2+-dependent, but the source of Ca2+ was different. Release of intracellular Ca2+ stores produced I1, whereas influx of extracellular Ca2+ produced I2; Ca2+-free bathing media and inorganic calcium channel blockers (Mn2+, Co2+) did not alter I1 but completely and reversibly inhibited I2. Injection of the Ins(1,4,5)P3 metabolite, inositol 1,3,4,5 tetrakisphosphate (Ins(1,3,4,5)P4) (0.2-10.0 pmol) generated a Ca2+-dependent Cl- current with superimposed current oscillations that resulted from release of intracellular Ca2+, not Ca2+ influx. Injection of the Ins(1,3,4,5)P4 metabolite, inositol 1,3,4-trisphosphate (10.0 pmol), or the synthetic inositol trisphosphate isomer, inositol 2,4,5-trisphosphate (1.0-10.0 pmol), mimicked the effect of Ins(1,4,5)P3, stimulating an I1 resulting from release of intracellular Ca2+ and an I2 resulting from influx of extracellular Ca2+. The results indicate that several inositol trisphosphate isomers stimulate both release of intracellular Ca2+ and influx of extracellular Ca2+. Ins(1,3,4,5)P4 also stimulated release of intracellular Ca2+, but it was neither sufficient nor required for Ca2+ influx. PMID- 2981052 TI - Cardboard chewing: cause and effect of iron-deficiency anemia. PMID- 2981053 TI - Muscle contraction and movement of cellular organelles: are there two different types of mechanisms for their generation? AB - It has been shown that myosin molecules attached to Covaspheres can "walk along" polar actin filament in vitro. The driving force for this movement seems to explain only about 1% of the isometric tension developed by a muscle fibre. Therefore, the driving force for the bead movement seems to be incompatible with that found in muscle, and the bead movement cannot be considered as a model for muscle contraction. The origin of the bead movement may be related to a "molecular jet" process, resulting from the rapid ejection of the MgATP splitting products. This "molecular jet" might also explain the movements of many cellular organelles. PMID- 2981054 TI - Metabolism of inositol-1,3,4,6-tetrakisphosphate to inositol pentakisphosphate in adrenal glomerulosa cells. AB - Angiotensin II stimulates rapid formation of inositol-1,4,5-trisphosphate (Ins 1,4,5-P3) in bovine adrenal glomerulosa cells. In addition to being rapidly metabolized to lower inositol phosphates, Ins-1,4,5-P3 is converted to Ins 1,3,4,5-tetrakisphosphate (Ins-1,3,4,5-P4) and Ins-1,3,4-P3 which is in turn phosphorylated to a further Ins-P4 isomer, namely Ins-1,3,4,6-P4. In bovine adrenocortical cytosol [3H]Ins-1,3,4,5-P4 and [3H]Ins-1,3,4-P3 were converted to Ins-1,3,4,6-P4 and inositol pentakisphosphate (Ins-P5) in a metabolic sequence suggesting that unlike Ins-1,3,4,5-P4, Ins-1,3,4,6-P4 is a direct precursor of Ins-P5. Consistent with this assumption, [3H]Ins-1,3,4,6-P4 was converted to Ins P5 in electropermeabilized adrenal glomerulosa cells. These findings demonstrate that Ins-1,3,4,6-P4 is an intermediate link between InsP3 metabolism and the higher inositol phosphates detected in several tissues. PMID- 2981055 TI - Argon laser treatment of radiation proctitis. PMID- 2981056 TI - Effect of bicarbonate feeding on intestinal absorption of calcium and vitamin D metabolism in rats. AB - The effect of bicarbonate (NaHCO3) feeding on intestinal absorption of calcium and phosphorus and vitamin D metabolism was investigated. Gut sac uptake of 45Ca and [32P]phosphate in vitro, as well as the plasma level of vitamin D metabolites, was determined in rats fed with NaHCO3 for 9 days and their pair-fed controls. Duodenal uptake of 45Ca and [32P]phosphate by alkalotic rats was less than duodenal uptake by controls. 1,25(OH)2D plasma level was much higher in alkalotic rats than in their controls, and 25(OH)D slightly higher than in controls. These data suggest that NaHCO3 feeding causes a substantial increase in 1,25(OH)2D plasma level, but interferes with calcium and phosphorus absorption in the duodenum. PMID- 2981057 TI - Eosinophils and bronchial inflammation. PMID- 2981058 TI - The precipitation of asthma by upper respiratory infections. AB - A number of important mechanisms have been identified by which viruses can provoke asthma. From the data available, there does not appear to be one single mechanism available to explain virus-induced asthma. The relationship between viral URIs and asthma is complex and involves many organ systems: airway epithelium, autonomic nervous system control, and the immediate hypersensitivity system. Identifying the effects of respiratory viruses on airway function remains an important undertaking as we try to better understand and control this precipitant of asthma. PMID- 2981059 TI - Leukotrienes and other lipid mediators of asthma. AB - It is now recognized that, in addition to the preformed mast cell granule mediators, newly generated lipid compounds are likely to be exceedingly important in the mediation of allergic asthma and other atopic diseases. That the initiating event in allergic diseases evokes a far more complex set of biochemical events than those that only lead directly to the release of histamine and other preformed mediators, and that the functional efficacies of the leukotrienes, PGD2, and PAF are significant for allergic pathobiology mandate that the latter compounds will necessarily be subject to efforts for future therapeutic intervention in allergic patient populations. PMID- 2981060 TI - Regulation of estrogen and progesterone receptor levels in mouse mammary epithelial cells grown in serum-free collagen gel cultures. AB - The effect of collagenase dissociation of virgin mouse mammary glands on the level of mammary epithelial cytosolic estrogen receptors (ER) and progesterone receptors (PR) was assessed. After cell dissociation, ER was present in mammary epithelial cells at concentrations similar to those found in the whole gland. However, PR appeared to be affected by the collagenase treatment. The regulation of ER and PR in mouse mammary epithelial cells isolated by collagenase dissociation and grown within collagen gels was then determined. After 7 days in culture under serum-free conditions inside a collagen gel, PR and, to a lesser extent ER, as characterized by high affinity binding and specificity, were present in the epithelial cells. Although at a low level, the ER were determined to be functional, since estradiol (E2) was able to promote nuclear accumulation of ER and to induce PR. PRL was able to increase cytosolic ER and PR concentrations. The combination of progesterone (P) and PRL was more effective than PRL or P alone in increasing PR. The induction of PR by P and PRL was inhibited when epidermal growth factor was present in the culture medium. Previous studies have shown that P, PRL, and epidermal growth factor, but not E2 (either alone or in combination with these factors) are able to stimulate cell proliferation in vitro. We conclude that the effects of E2 on protein synthesis and proliferation are dissociated in vitro. The difference between the effect of E2 and PRL or P on growth may be related either to the initial concentrations of their respective receptors or estrogen may stimulate growth indirectly. PMID- 2981061 TI - Characterization of adrenocorticotropin receptors that appear when 3T3-L1 cells differentiate into adipocytes. AB - The binding of an 125I-labeled analog of ACTH, [125I]Tyr23,Phe2,Nle4-ACTH-(1-38), to differentiated 3T3-L1 fat cells was characterized. Time-dependent binding, which was inhibited by saturating concentrations of unlabeled ACTH (0.44 microM), could be demonstrated in the differentiated cells. Using 0.4 nM [125I]ACTH analog and increasing concentrations of ACTH, the half-maximal concentration for inhibition by ACTH was 4.3 nM. Scatchard analysis demonstrated a single class of ACTH binding. There were approximately 3500 binding sites/cell. The binding of [125I]ACTH analog was specific in that it could be displaced by ACTH, ACTH-(1 19), ACTH-(1-17), and N-acetyl-Ser1-ACTH, but not by high concentrations of insulin, beta-endorphin, or polylysine. There was an excellent correlation between the ability of ACTH and its analogs to inhibit [125I]ACTH analog binding and the ability of ACTH and its analogs to stimulate cAMP production. In contrast, no saturable binding could be demonstrated when undifferentiated 3T3-L1 fibroblasts, which are not responsive to ACTH, were studied. Thus, differentiation of 3T3-L1 cells into the adipocyte form is accompanied by the appearance of receptors for ACTH. These receptors allow the adipocytes to respond to ACTH. PMID- 2981062 TI - Alpha-melanocyte-stimulating hormone stimulation of aldosterone secretion in hypophysectomized rats. AB - Intact and hypophysectomized male rats on low and high sodium diets were treated with sc infused ACTH and alpha MSH, and the levels of aldosterone and corticosterone were determined in truncal blood. Plasma aldosterone levels were lower in hypophysectomized animals on the low sodium diet than in intact animals on the low sodium diet. Alpha MSH (8 micrograms/day) restored plasma levels of aldosterone to normal in hypophysectomized rats on the low sodium diet. ACTH (6 micrograms/day) did not cause significant changes in plasma levels of aldosterone in hypophysectomized animals. ACTH restored plasma corticosterone to normal in hypophysectomized rats, whereas alpha MSH had no effect on corticosterone. Alpha MSH did not increase plasma aldosterone levels in intact rats. These data suggest that alpha MSH may be important in the regulation of aldosterone secretion in the rat and that zona glomerulosa responsiveness to alpha MSH in vivo is increased by hypophysectomy. The mechanisms of alpha MSH action on glomerulosa cells are different from those of ACTH. PMID- 2981063 TI - Differences in the lateral mobility of receptors for luteinizing hormone (LH) in the luteal cell plasma membrane when occupied by ovine LH versus human chorionic gonadotropin. AB - Receptors for LH are internalized by ovine luteal cells 50 times slower when occupied by hCG than when occupied by ovine LH (oLH). To determine if differences in the rate of internalization were due to differences in the lateral mobility of the hormone-receptor complexes in the cell membrane, the diffusion coefficients of oLH- and hCG-LH receptor complexes were measured using fluorescence photobleaching recovery methods. Tetramethylrhodamine isothiocyanate (TRITC) labeled oLH and hCG, which retained full ability to bind to receptor, were bound to LH receptors on enzymatically dispersed ovine luteal cells. Molecules labeled with TRITC within a 3-micron 2 region of the cell surface were bleached by a 500 msec pulse of 3 mW laser light at a wavelength of 514.5 nm. The laser beam intensity was then attenuated 20,000-fold, and fluorescence from the bleached area was measured by single photon counting as unbleached fluorescent hormone receptor complexes diffused into the region. Data were analyzed on-line by a NOVA 3/12 computer. The oLH-LH receptor complex had a diffusion coefficient of 1.9 +/- 1.0 X 10(-10) cm2/sec-1, a value comparable to that of cell surface proteins nonspecifically labeled with succinylated Concanavalin A. Fluorescence recovery after photobleaching was 35%. In contrast, hCG-LH receptor complexes were immobile on the time scale of the experiment, implying that the diffusion coefficient was substantially less than 1 X 10(-11) cm2/sec-1. Deglycosylated hCG TRITC bound to LH receptor had a diffusion coefficient (1.1 +/- 0.1 X 10(-10) cm2/sec-1) similar to that of receptors occupied by oLH. Thus, it appears that the carbohydrate portion of the hCG molecule plays a role in decreasing the mobility of the receptor for LH. These data demonstrate that the rate of lateral movement of the LH receptor in the plasma membrane of luteal cells appears to be modulated by the nature of the bound hormone. PMID- 2981064 TI - Epidermal growth factor receptors in isolated adult mouse intestinal cells: studies in vivo and in organ culture. AB - In isolated intestinal cells from adult fed mouse, the binding of [125I]epidermal growth factor (EGF) was time and temperature dependent. Maximum binding was obtained after 30 min of incubation at 20 C. For a concentration of 5 X 10(-11)M [125I]EGF (180 microCi/micrograms), specific binding for isolated cells from duodenum, jejunum, and ileum was closely similar, with means of 9.4 +/- 0.9, 13.3 +/- 0.8, and 9.3 +/- 2.0%/mg protein, respectively. The binding increases along the crypt-villus axis. Inhibition dose-response analysis indicated high affinity binding with 50% inhibition at 3 X 10(-10) M unlabeled EGF. The specific binding decreased by 19% after 48 h of fasting. In the P1 fraction (microsome and lateral membranes) from scrapings or isolated cells of the jejunal mucosa, specific binding was 2.4 +/- 0.8 and 6.5 +/- 0.7%/mg protein, respectively. In the P2 fraction (brush border), specific binding was 6.9 +/- 1.6% and 16.3 +/- 0.7%/mg protein. After 24 h of organ culture, specific binding is not modified in duodenal explants. Moreover, in the presence of EGF (500 ng/ml) in the culture medium, the binding is decreased by 72%. These results show that isolated intestinal cells from adult mice possess a high concentration of EGF receptors that exhibit kinetic properties identical to those of other EGF target cells. Unlike insulin receptors, EGF receptors are numerous on the intestinal brush border and in intestinal crypts and decrease by fasting. PMID- 2981066 TI - Effects of gonadotropin-releasing hormone receptor blockade on rat testicular gonadotropin and lactogen receptors, steroidogenesis, and responses to human chorionic gonadotropin stimulation. AB - Testicular endocrine regulation was studied in adult male rats after treatment with a potent GnRH antagonist analog [N-Ac-D-p-Cl-Phe1,2, D-Trp3, D-Lys6, D-Ala10 GnRH (Ant.)] given in doses of 1 mg/kg at 0, 12, and 24 h. One group of animals also received an injection of human CG (hCG) (600 IU/kg) at 12 h, and all animals were killed at 36 h for hormone and receptor (R) measurements. Treatment with Ant. blocked greater than 95% of the pituitary and testicular GnRH-R, and decreased serum LH concentration by greater than 90%. Testicular lactogen-R content was decreased by 60% (P less than 0.01), but there was no change in LH-R and FSH-R concentrations. Ant. decreased serum and testicular testosterone levels by 90%, and testicular capacity to produce testosterone in vitro by 50% (P less than 0.01). No decrease was observed in production rates of cAMP and progesterone. hCG alone abolished testicular LH-R, decreased lactogen-R by 55% (P less than 0.01), and GnRH-R by 65% (P less than 0.01). Desensitization of cAMP and testosterone production, and an increase in progesterone-testosterone ratio, were seen after hCG. hCG + Ant. treatment resulted in R, cAMP, and steroid responses that were indistinguishable from those seen after hCG alone. These findings indicate that: 1) Ant.-induced hypogonadotropism decreases testicular lactogen-R concentration and testosterone production; 2) testicular GnRH-R and lactogen-R are subject to heterologous down-regulation by hCG; and 3) inhibition of the putative GnRH-mediated regulation of testis by Ant. blockade of the GnRH-R does not change testicular response to hCG-treatment in vivo. Hence, the present observations still leave the physiological role of testicular GnRH-R open. PMID- 2981065 TI - Arachidonate stimulates prolactin release in vitro: a role for the fatty acid and its metabolites as intracellular regulator(s) in mammotrophs. AB - We investigated the involvement of arachidonate in the PRL secretory process using three experimental systems: hemipituitary glands incubated in vitro, cultured pituitary cells, and dispersed anterior pituitary cells perifused in columns. Arachidonate (100 microM) significantly (P less than 0.05) stimulated PRL release in the former system and stimulated PRL secretion in a dose-related manner in cultured cells. In hemipituitary glands, indomethacin, a cyclooxygenase inhibitor, potentiated the arachidonate-mediated stimulation, whereas nordihydroguaiaretic acid or BW755c abolished it. The latter two agents, but not indomethacin, abolished the effect of phospholipase A2 on PRL release in vitro. BW755c also inhibited the stimulatory effect of TRH on PRL release in both experimental systems. Conversely, the stimulation of PRL release by phorbol myristate acetate (PMA), although significantly reduced, was not abolished by either nordihydroguaiaretic acid or BW755c. Quinacrine, a phospholipase A2 inhibitor, also abolished the stimulatory effect of phospholipase A2 or TRH on PRL release. In cultured cells, quinacrine inhibits basal PRL release, but does not affect PRL release induced by arachidonate or (Bu)2 cAMP. These results more firmly establish a role for arachidonate as an intracellular mediator of PRL release and suggest the involvement of an arachidonate metabolic pathway(s) (lipoxygenase and epoxygenase) other than prostaglandin or thromboxane formation. The effect of PMA on PRL release may be attributable only in part to an increase in the production of arachidonate metabolites, and most of PMA's effect on PRL release may relate to its activation of protein kinase C. PMID- 2981067 TI - Adenosine 3',5'-monophosphate-dependent phosphoproteins in human placenta. AB - cAMP modulates estrogen, hCG, and lactate syntheses by the human placenta. The major effects of cAMP are presumably mediated through the phosphorylation of specific regulatory phosphoproteins after cAMP activation of cAMP-dependent protein kinase. cAMP-dependent phosphoproteins have not been identified in the placenta. Homogenates and cytosol from term human placenta were subjected to an endogenous protein phosphorylation assay using [gamma-32P]ATP in the presence and absence of 1.0 microM cAMP. Protein phosphorylation was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. cAMP stimulated 32P incorporation into proteins with mol wt of 25,000, 27,000, 39,000, 45,000, 52,000, 58,000, and 73,000 (P less than 0.02). Half-maximal 32P incorporation was observed with 1.0 X 10(-7) M cAMP, which was similar to the concentration required for half-maximal histone kinase activity (8.5 +/- 2.9 X 10(-8) M). cGMP induced 32P incorporation into the same placental proteins as cAMP, but 2 orders of magnitude greater cGMP concentrations were required to achieve phosphorylation levels similar to those caused by cAMP. cAMP-dependent protein kinase inhibitor completely blocked cGMP-induced phosphorylation, even when histone protein was added. Therefore, no evidence of a cGMP-dependent protein kinase or specific cGMP-dependent phosphoproteins were detected. CaCl2 (10(-8) - 10(-4) M) had no effect on cAMP-induced 32P incorporation into the seven cAMP-dependent phosphoproteins. However calcium induced 32P incorporation into four other proteins (mol wt, 97,000, 90,000, 20,000, and 19,000). Regulation of placental metabolism by catecholamines and other hormones known to mediate intracellular cAMP or calcium levels may be accomplished by phosphorylation of these phosphoproteins. PMID- 2981068 TI - Gonadotropic regulation of aromatase activity in the adult rat testis. AB - Aromatase activity during gonadotropin action in vivo and in vitro was examined in purified Leydig cells to further define the early effects of LH and for elucidation of the enzymatic processes involved in the development of late lesions of androgen biosynthetic pathway. Aromatase was measured by the tritiated water release method using [1 beta-3H]testosterone as substrate. The enzyme activity was proportional to the amount of cells (0.1-1.0 X 10(6] incubated, increased with the incubation time (2-60 min), was inhibited by androstatriendione (ED50, 5.0 microM), and showed a Km for testosterone of 1.69 microM. Aromatase activity was stimulated (10-20%; P less than 0.05) 1 h after treatment of rats with a single sc dose of 5 micrograms hCG. This activation preceded the late steroidogenic lesion at the site of 17 alpha-hydroxylase and 17,20-desmolase activity by 2-5 h. A RIA of improved sensitivity (0.5 pg) was developed to detect the very low cellular and secretory levels of estradiol. The testicular contents of testosterone and estradiol showed small increases (P less than 0.05) within 40 and 60 min after hCG treatment, respectively. Testicular testosterone levels reached a peak by 1 h after injection and preceded the peak level of estradiol formation by 2 h. After in vitro treatment of cultured Leydig cells with 100 ng hCG, the aromatase activity was significantly increased within 30 min (P less than 0.05) and then returned to control levels for up to 16 h of culture. A similar temporal pattern for enzyme activation was observed after treatment of cultures with 8-bromo-cAMP or forskolin (23-27% above control; P less than 0.05), while cholera toxin stimulated aromatase activity at 2 h. Net testosterone accumulation in the incubation medium increased 30 min after the hCG treatment and reached a plateau by 4 h. A small but significant increase in estradiol levels (P less than 0.05) was also observed at 30 min, remaining constant until 120 min, which was followed by a sharp rise parallel to that of testosterone. These results suggest that the estradiol-mediated desensitization of the Leydig cell observed after hCG administration is consistent with an early cAMP-dependent activation of aromatase and a further rise in estradiol formation due to increased substrate availability. PMID- 2981069 TI - Conversion of estrogen receptor from a state with low affinity for estradiol into a state of higher affinity does not require 4S to 5S dimerization. AB - This study was undertaken to establish whether the heat-promoted conversion of receptor-estradiol complex (RE2) from a state with fast into a state with slow E2 dissociation requires 8S/4S to 5S transformation. The calf uterine estrogen receptor labeled with [3H]E2 at 0 C (state with low affinity for E2) was immobilized on hydroxylapatite (HAP) in the absence (8S oligomer) or presence (4S monomer) of 0.4 M KCl and heated at 28 C in the presence of unlabeled diethylstilbestrol. Under these conditions, the dissociation of [3H]E2 was biphasic and occurred at rates similar to those obtained with R[3H]E2 free in cytosol. In contrast to the latter, however, the heat-promoted conversion of HAP immobilized R[3H]E2 into a state of higher affinity for E2 was not accompanied by receptor dimerization, since the HAP eluate (0.4 M phosphate buffer) contained only the 4S monomer; upon reheating or desalting of this eluate, 4S to 5S dimerization occurred. The heat-promoted formation of 4S RE2 monomers with higher affinity for E2 was not due to monomer-HAP interactions, since even after elution from HAP, the 4S RE2 remained in the state of higher affinity for E2. Addition of pyridoxal 5'-phosphate to slow dissociating, high affinity 5S R[3H]E2 dimers free in cytosol induced rapid [3H]E2 dissociation, although the receptor remained unaltered in the transformed dimerized state. The effect of PLP was readily reversed by the addition of lysine. It is proposed that the 4S receptor monomers exist in two conformational states; upon E2 binding to the low affinity state, conformational changes result in stronger interactions between the steroid and the amino acid residues of the estrogen-binding domain; thus, the rate of E2 dissociation decreases. The formation of this 4S RE2 state with higher affinity for E2 is independent from receptor dimerization. A model is presented in which 4S to 5S transformation may lead to stabilization of 4S monomers in the conformation with higher affinity for E2. PMID- 2981070 TI - Secreted mouse prolactin (PRL) and stored ovine PRL. II. Role of amides in receptor binding and immunoreactivity. AB - Native PRL and des-amido forms 1, 2, and 3 were tested for their individual immunochemical and receptor-binding abilities. The results show that deamidation of either secreted mouse PRL or stored ovine PRL alters their binding in a radioreceptor assay. For each accumulated deamidation there was a statistically significant (P less than 0.05) decrease in the binding potency of either stored ovine PRL or secreted mouse PRL to a cell membrane receptor preparation. RIAs indicated that there was a statistically significant (P less than 0.05) decrease in PRL's immunopotency toward polyclonal antisera only when select residues deamidated. This study suggests that all the asn/gln residues which deamidate in order to make des-amido forms 1, 2, and 3 constitute part of the receptor-binding domains of both secreted and stored PRLs, while only a fraction of those same residues form portions of PRL's antigenic sites. Thus PRL's receptor-binding surface is separated from its antigenic sites, with only partial overlap being indicated. Our data also indicate that there are no major structural differences in the receptor-binding domains of secreted and stored PRLs. We also see that the receptor-binding domain of PRL has been highly conserved throughout evolution. Since the binding affinity of PRL to a membrane-bound receptor can be altered through deamidation, we view the process as a plausible regulatory mechanism for controlling the quantitative action of PRL at a given target organ. PMID- 2981072 TI - Cellular localization of rat testicular aromatase activity during development. AB - The aromatase activity from purified testicular sources (Leydig and Sertoli cells) of immature (5- and 15-day-old) and adult rats (60-day-old) was investigated by the tritiated water release method in isolated Leydig and Sertoli cells that were morphologically and functionally characterized. Electron micrographs of Sertoli cell preparations from different ages showed no marked changes, except that tight junctions between Sertoli cells normally present in 60 day-old rats were not observed in 5-day-old and rarely found in 15-day-old animals. Leydig cells underwent ultrastructural changes along with development, such as the appearance of thicker nuclear heterochromatin and laminar-like mitochondria. The 15-day-old rat interstitial tissue possessed less than 10% of Leydig cells morphologically similar to those present in the adult, whereas the rest were probably transition cells, since they did not show typical Leydig cell structure but were able to bind [125I]iodo-hCG, as evaluated by autoradiography. The number of LH/hCG-binding sites increased with age in Leydig cells, but was not detectable in Sertoli cells. The highest number of FSH-binding sites in Sertoli cells was observed in the 15-day-old animals. Minor FSH binding was found in Leydig cell preparations, which was consistent with the known LH contamination of the human FSH tracer preparation. cAMP production increased significantly in Leydig cells only after hCG treatment and in Sertoli cells after FSH stimulation. Both types of cells were shown to have the capacity for aromatization. The aromatase activity increased in the Leydig cell but decreased in the Sertoli cell during testicular development. The highest aromatase activity was found in adult rat Leydig cells, and the enzyme activity was significantly higher (2-fold) in purified Leydig cells than in crude interstitial cell preparations. Estradiol production in response to hCG stimulation in vitro was not different from the basal value in 5-day-old rat Leydig cells, but increased significantly in 60-day old rat Leydig cells. In conclusion, 1) Leydig cells are the major site of estrogen synthesis in adult rat testis; and 2) the low aromatase activity observed in immature rat Leydig cells could partially explain the differential response of the mature and immature rat testis to hCG-induced desensitization. PMID- 2981071 TI - Intracellular kinetics of free catalytic units dissociated from adenosine 3',5' monophosphate-dependent protein kinase in adrenocortical tumor cells (Y-1). AB - To define the role of cAMP in the actions of ACTH, the dissociation of cAMP dependent protein kinase and the subsequent intracellular location of its free catalytic units were monitored after exposure of Y-1 cells to ACTH, FSH, or cyclic nucleotide analog. To accomplish this, a fluorescinated cytochemical probe was used that complexes specifically with free catalytic units from cAMP dependent protein kinase. Also, the effects of hormone or nucleotide on secretion of fluorogenic steroids and DNA synthesis were examined. Y-1 cells dissociated protein kinase in a dose-dependent fashion when exposed to ACTH or cAMP analog, but did not respond to FSH, which was one of the control agents used. After 30 min of treatment with 1.5 X 10(-10) M ACTH, free catalytic units were observed only in the cytoplasm of Y-1 cells, whereas a similar time of exposure to 3 X 10( 10) M ACTH led to the appearance of catalytic units in nucleolus as well as in cytoplasm. ACTH (6 X 10(-10) M) caused a rise in cytoplasmic and nucleolar protein kinase dissociation proportionally greater than that seen in cultures exposed to 3 X 10(-10) M ACTH. Upon treatment with 6 X 10(-10) M ACTH, the amount of free catalytic units in cytoplasm and nucleolus was detectably greater than that in controls within 1 min of stimulation and continued to rise with increasing time of exposure to hormone. The nuclear, mostly nucleolar, content of free catalytic unit appeared to peak after 15 min of stimulation, while cytoplasmic enzyme levels continued to rise up to 60 min. Exposure of Y-1 cells to nucleotide analog caused cAMP-dependent protein kinase dissociation with temporal kinetics and a subcellular distribution similar to that seen after ACTH stimulation. We conclude that actions of ACTH are mediated by cAMP-dependent protein kinases. Further, there appear to be two intracellular pools of protein kinase, one nucleolar, the other cytoplasmic, and these may be independently regulated, with the nucleolar enzyme requiring higher concentrations of ACTH for dissociation than those needed for cytoplasm protein kinase. These observations may be relevant to the fact that more ACTH is required to inhibit DNA synthesis than is necessary to enhance steroid production. PMID- 2981073 TI - Identification and characterization of arginine vasopressin receptors in the rat testis. AB - We have previously shown that arginine vasopressin (AVP) directly inhibits testicular steroidogenesis in vitro. In the present study, binding of neurohypophysial peptides to interstitial cells of the rat testis was studied using [3H]AVP as the ligand. Interstitial cells were obtained from adult rat testis after collagenase dispersion and were incubated with [3H]AVP in the presence or absence of unlabeled AVP. Binding equilibrium was reached by 60 min at 4 C, while incubation at higher temperatures (23 and 37 C) resulted in an apparent decrease in binding. Scatchard plot analysis of equilibrium binding data revealed the existence of one class of high affinity, low capacity binding sites (Kd = 1.0 +/- 0.3 nM; maximal binding = 8.5 fmol/10(6) cells). In addition, the rate constants of association and dissociation were calculated to be 0.024 nM-1 min-1 and 0.009 min-1, respectively. Addition of naturally occurring neurohypophysial hormones as well as their synthetic analogs inhibited [3H]AVP binding to testis cells, resulting in parallel displacement curves. The order of potencies for the native peptides was: AVP = lysine vasopressin = arginine vasotocin (IC50, 5 X 10(-10) M) greater than oxytocin = mesotocin (IC50, 4 X 10( 7) M) greater than isotocin = glumitocin (IC50 greater than 10(-6) M). Furthermore, two potent vasopressor antagonists, d(CH2)5Tyr(Me)AVP ([1-(beta mercapto-beta, beta-cyclopentamethylenepropionic acid), 2-(O-methyl)tyrosine]AVP) and dPTyr(Me)AVP ([1-deaminopenicillamine-2-(O-methyl)tyrosine]AVP) competed for [3H]AVP binding with a higher affinity (IC50, approximately 10(-11) M) than native AVP. In contrast, a selective antidiuretic agonist, dDAVP (1-deamino-8-D AVP), only competed weakly for receptor binding, while a specific oxytocic agonist, (Thr4,Gly7)oxytocin, did not affect AVP binding. These results suggested that the testis may contain the V1 receptor subtype. Studies on the intratesticular distribution of AVP receptors indicated minimal binding to cells derived from the seminiferous tubule, while most of the AVP-binding sites sediment with enriched fractions of Leydig cells after Metrizamide density gradient centrifugation. AVP-binding sites were also found in rat liver, kidney, and anterior pituitary (10.7, 2.6, and 1.7 fmol/mg protein), whereas adrenal, cerebellum, prostate, and hypothalamus were devoid of AVP-binding sites. Thus, we have demonstrated the presence of high affinity, stereospecific receptors for AVP in the interstitial cell compartment of the rat testis. These V1 receptors may mediate the direct inhibitory action of neurohypophysial hormones on testicular Leydig cell steroidogenesis. PMID- 2981074 TI - Quantification of morphological changes in pituitary corticotropes produced by in vivo corticotropin-releasing factor stimulation and adrenalectomy. AB - Physiological and morphological changes produced by corticotropin-releasing factor (CRF) stimulation in vivo were studied in pituitaries immunocytochemically stained for ACTH. After 48-h ip minipump infusions of 10 or 50 ng/min CRF, serum ACTH levels were increased significantly over values in control groups that included both intact rats and rats exposed to sham abdominal surgery. Correlative morphological changes included a striking increase in corticotrope cell area. This was coupled with an apparent decrease in the percentage of stained cells, probably due to degranulation. The cellular responses were similar to those after adrenalectomy described previously by us and others. Therefore, in a parallel study, additional groups of rats were adrenalectomized and studied 24 and 48 h after the surgery. Even greater changes in serum ACTH, corticotrope cell area, and percentages were observed in the adrenalectomized rats. The difference between the CRF-infused and adrenalectomized groups was probably due to the lack of corticosterone feedback in the latter group. Among the control groups, there were no differences between intact rats and rats exposed to sham abdominal surgery. Rats subjected to sham adrenalectomy, however, showed corticotrope responses similar to those of CRF-infused rats, except that the cells were more densely stained. The present studies thus show dramatic changes in ACTH cell area, extent of staining, and percentages after in vivo CRF stimulation. In all of the experimental groups, an excellent correlation existed between serum ACTH levels and the degree of the morphological changes in the corticotropes. PMID- 2981075 TI - EGF receptor antiserum inhibits bone resorbing activity produced by a rat Leydig cell tumor associated with the humoral hypercalcemia of malignancy. AB - The humoral hypercalcemia of malignancy (HHM) is a syndrome caused by tumor cells releasing unknown circulating factors which stimulate osteoclastic bone resorption. In the D6 variant of the rat Leydig cell tumor model of HHM, we found that tumor extracts and tumor cell conditioned medium contained a macromolecular bone resorbing factor which coeluted on column chromatography with transforming growth factor activity (TGF). This observation led to the hypothesis that the tumor-derived bone resorbing factor was a TGF which interacts with the epidermal growth factor (EGF) receptor. To test this hypothesis, we examined the effects of two classes of antisera to the EGF receptor on bone resorption stimulated by conditioned medium from Leydig D6 tumor cells using organ cultures of fetal rat long bones. The antiserum which blocks the binding of EGF to its receptor inhibited bone resorption stimulated by tumor conditioned medium and by EGF. The second antiserum to the EGF receptor which does not block EGF binding or biological activity had no effect on bone resorption stimulated by either tumor conditioned medium or EGF. Neither antiserum had any effect on bone resorption stimulated by parathyroid hormone (PTH). These results indicate that the tumor derived bone resorbing factor is dependent upon the availability of EGF receptors for its activity and are consistent with it being a TGF. PMID- 2981076 TI - Possible delta receptor mediation of the effect of beta-endorphin on luteinizing hormone (LH) release, but not on prolactin (PRL) release, in the ovariectomized rat. AB - Administration of opioid receptor antagonists was utilized to determine the opioid receptor type involved in the suppression of LH release by beta-endorphin (beta-END). Long-term (three to four weeks) ovariectomized rats with chronic third ventricular cannulae were fitted with jugular catheters and received treatment with vehicle or one of three opioid antagonists. The delta antagonist ICI 154, 129, but not the mu1 or mu antagonists naloxazone or beta funaltrexamine, respectively, blocked the suppressive effect of beta-END on plasma LH levels and transiently but significantly increased LH levels above preinfusion value. None of the antagonists significantly reduced the beta-END induced release of PRL. These results provide evidence that the inhibitory effect of beta-END on LH release may be mediated by delta receptors. PMID- 2981077 TI - Enhanced follicle-stimulating hormone activity of deglycosylated human chorionic gonadotropin in ovarian granulosa cells. AB - The receptor binding properties and biological actions of chemically deglycosylated asialo human CG (AHF-hCG) were studied in ovarian granulosa cells from diethylstilbestrol (DES)-treated immature rats. In ovarian homogenates from DES- and FSH-treated rats, the relative binding affinity of AHF-hCG was 2-fold higher than that of native hCG and 14-fold higher than that of ovine LH. When assayed for LH-like activity in granulosa cells from DES plus FSH-treated animals, the deglycosylated hormone behaved as a partial agonist in terms of cAMP formation, but fully stimulated progesterone production to the same extent as that elicited by LH. When added with LH to FSH-treated cells, AHF-hCG inhibited LH-stimulated cAMP formation by 70% but did not alter the elevated level of progesterone production. These findings are consistent with the presence of excess or spare LH receptors in the maturing granulosa cell. When added to freshly prepared granulosa cells which have minimal LH receptors, AHF-hCG decreased FSH-stimulated cAMP production by 20% and reduced progesterone production by 50% and increased cGMP formation by 100% during 48 h of culture. The ability of AHF-hCG to decrease the progesterone response to FSH suggests that no spare FSH receptors are present during granulosa cell differentiation. In contrast, native hCG did not alter FSH-induced cAMP or progesterone production but reduced the cGMP responses to FSH and choleragen. Whereas native hCG displayed negligible binding potency when compared with that of ovine FSH in competition with [125I]iodo-human FSH for ovarian receptors, AHF-hCG bound to FSH receptors with about 5% of the binding affinity of ovine FSH. In choleragen treated granulosa cells, the increases in cAMP and progesterone synthesis were enhanced by addition of both hCG and AHF-hCG, and cGMP production was increased by AHF-hCG but slightly decreased by hCG. These results indicate that the enhanced LH receptor affinity caused by removal of the sugar moieties from hCG is accompanied by a relatively greater increase in FSH receptor affinity, and that deglycosylated hCG acts as a partial agonist with the ability to modify granulosa cell responses to both LH and FSH. PMID- 2981078 TI - Putative nuclear triiodothyronine receptors in tadpole erythrocytes during metamorphic climax. AB - The characteristics of the nuclear T3 receptors present in red blood cells (RBCs) of Rana catesbeiana tadpoles undergoing metamorphic climax have been investigated with a T3 saturation technique. Because there were significant amounts of receptor-bound endogenous hormone, prolonged incubation in vitro (48 h at 21 C) was necessary to achieve binding equilibrium. Receptor number, which averaged 783 +/- 35 (+/- SE) sites/nucleus during early prometamorphosis (stages XIV-XVI), increased rapidly during the subsequent stages of this phase. By stage XIX, receptor number had reached a maximum of 2464 +/- 152. During climax, receptor number decreased steadily, and by stage XXV, it was lower than that observed during premetamorphosis (stages V-X). This decrease was attributed to the replacement of larval RBCs, which have a high receptor content, by adult RBCs, which possess significantly fewer sites per nucleus. At midclimax, adult and larval RBCs were separated on a Renografin continuous density gradient. Adult cells constituted more than 65% of the total cell population and were shown to contain only 126 +/- 46 sites/nucleus. Although the striking increase in receptor number preceded the substantial increases in plasma T4 and T3 levels that occurred during climax, it was associated with a measurable increase in plasma thyroid hormone levels. Furthermore, receptor number in stage XIV tadpoles was increased markedly after immersion for 14 days in water containing sufficient T3 to raise plasma T3 only to levels normally observed in late prometamorphosis. These findings strongly support the hypothesis that the increases in receptor number and plasma thyroid hormone levels that occur before climax in this species are causally related. PMID- 2981079 TI - Cytomegalovirus colitis. Report of the clinical, endoscopic, and pathologic findings in two patients with the acquired immune deficiency syndrome. AB - We report 2 homosexual patients with the acquired immune deficiency syndrome and histopathologic evidence for cytomegalovirus colitis. In each case, the initial endoscopic impression was Kaposi's sarcoma but the appropriate diagnosis was made by histologic demonstration of a cytomegalovirus vasculitis. Clinical manifestations may include diarrhea, abdominal pain, and hematochezia. Fulminant progression to gangrenous bowel was documented in 1 patient and was associated with histologic evidence of severe cytomegalovirus vasculitis. The pertinent clinical, endoscopic, and pathologic findings are reviewed. PMID- 2981080 TI - Hepatocellular carcinoma: an unusual course with hyperthyroidism and inappropriate thyroid-stimulating hormone production. AB - A 61-yr-old woman was found to have multicentric hepatocellular carcinoma associated with clinical hyperthyroidism. Marked elevations in serum thyroid stimulating hormone, T4, T3, and free T4 were documented. These values increased with tumor progression. It is postulated that this patient's hepatoma produced a substance stimulating thyroid-stimulating hormone synthesis and secretion with resultant inappropriate thyroid-stimulating hormone production and thyrotoxicosis. This unusual association of hepatocellular carcinoma with clinical and biochemical evidence of hyperthyroidism and inappropriate thyroid stimulating hormone production has not been previously reported. PMID- 2981081 TI - Role of the liver in the degradation of lipoproteins. PMID- 2981082 TI - Cysteamine and propionitrile inhibit the rise of duodenal mucosal alkaline secretion in response to luminal acid in rats. AB - Effects of subulcerogenic doses of cysteamine (100 mg/kg s.c.) and propionitrile (5 mg/kg) on alkaline secretion by duodenal surface epithelium and pH at the surface of this mucosa were assessed in duodenum of anesthetized rats. Alkaline secretion was titrated in situ, using segments of duodenum just distal to the Brunner's glands area and devoid of pancreatic HCO3-. Surface pH was measured by advancing pH-sensitive microelectrodes from the luminal solution to the epithelial cell surface. Proximal duodenum from bullfrogs was used to study effects of cysteamine on alkaline secretion in vitro. Cysteamine caused an increase in alkaline secretion in the rat during the first hour after administration, but rates after 5 and 20 h were the same as in controls and cysteamine (1 mg/ml) had no effect on secretion in vitro. Neither in vitro nor in vivo did cysteamine affect the rise in alkaline secretion in response to exogenous prostaglandin E2 (and dibutyryl-cyclic adenosine monophosphate). Luminal acid is a potent stimulant of duodenal mucosal alkaline secretion. By delayed (5 h) actions, both cysteamine and propionitrile inhibited the rise in alkaline secretion in response to a 5-min exposure to luminal acid with pH 2.00 in the rat. Cysteamine also depressed the ability of this mucosa to maintain a high rate of alkaline secretion during sustained exposure at pH 2.00 but had no such effect at pH 5.00. The former resulted in acidification of the pH gradient at the mucosal surface. Cysteamine is thus probably without effect on the HCO3- secretory process itself but impairs the ability of the duodenal mucosa to respond to acid. Inhibition of mechanisms mediating this response may contribute to the duodenal ulcerogenic actions of cysteamine and propionitrile. PMID- 2981083 TI - Increased adrenocorticotropin, cortisol, and arginine vasopressin secretion in primates after the antiglucocorticoid steroid RU 486: dose response relationships. AB - The antiglucocorticoid steroid, RU 486, elevated plasma ACTH concentrations when administered im at 0700 h in a dose of 1.0 mg/kg in nonhuman primates (Macaca fascicularis; P less than 0.05). The duration, but not the magnitude, of this response increased after 5.0 mg/kg RU 486 im; release of ACTH did not rise further after 10 mg/kg. Peak ACTH elevations occurred 1-2 h after RU 486 administration. Plasma cortisol concentrations peaked 4 h after RU 486 administration and the response was significant only after 5.0 mg/kg RU 486. Plasma arginine vasopressin (AVP) concentrations also increased after RU 486 but the increase occurred only after the 10.0 mg/kg dose (P less than 0.05). The AVP elevation was greatest 4 h after 10 mg/kg RU 486 and was abolished by dexamethasone pretreatment. We conclude that: 1) RU 486 elevates plasma ACTH, cortisol, and AVP concentrations in a manner which is both dose and time dependent, 2) ACTH release occurred at an order of magnitude lower dose than did AVP release, and 3) plasma AVP changes after RU 486 are glucocorticoid dependent. PMID- 2981085 TI - Oxytocin receptors in nonpregnant human uterus. AB - The concentration and distribution of specific [3H]oxytocin-binding sites in the nonpregnant human uterus were studied. Oxytocin was bound with an apparent Kd of about 1 nM in the crude membrane fractions of the fundus and corpus. A second class of sites with lower affinity and higher capacity for oxytocin also was found. Consistent with its high uterotonic potency in nonpregnant uteri, arginine vasopressin was as effective as oxytocin in inhibiting [3H]oxytocin binding to corpus and fundus membrane preparations. The concentrations of high affinity binding sites in fundus and corpus were similar and were significantly higher than those of membrane fractions from isthmus or Fallopian tubes. The lowest concentration of sites was found in the cervix. Endometrial membrane preparations contained oxytocin-binding sites in about the same concentration as that in the myometrium. The concentrations of oxytocin receptors in all parts of the nonpregnant uterus were somewhat higher in the luteal phase than in the follicular phase. Concentrations were lowest in postmenopausal uteri. The concentrations of oxytocin receptors in nonpregnant uteri were 50-100 times lower than those in uteri near the end of gestation. These differences correspond to the differences in sensitivity to oxytocin between luteal phase and follicular phase and between the nonpregnant and late term uterus. These findings add support to the evidence that the binding sites for oxytocin represent true oxytocin receptors. PMID- 2981084 TI - The effects of estrogen and progestin on endogenous opioid activity in oophorectomized women. AB - Sex steroids may modulate the secretion of beta-endorphin (beta-EP). Naloxone (Nal), an opioid antagonist, has been used as a probe of central opioid activity. Nal-evoked responses of PRL and LH were evaluated in the midluteal (ML) and late follicular (LF) phases of ovulatory women (Pre) and compared to responses of oophorectomized women before and after the administration of conjugated estrogens (CE) and again after CE and progestin administration. In the ML and LF phases, serum LH increased significantly (P less than 0.05 and P less than 0.01, respectively) during Nal infusion for 4 h, while PRL did not change. In oophorectomized women, there were no significant changes in LH or PRL during Nal infusion. After 3 weeks of CE treatment (1.25 mg daily), LH increased during Nal infusion (P less than 0.05), as did PRL (P less than 0.01). After treatment with CE and medroxyprogesterone acetate (MPA), LH and PRL both increased (P less than 0.05 and P less than 0.01, respectively). The area under the LH curve during Nal infusion after CE and MPA treatment was greater than that after CE alone. Both of these responses were comparable to those of the LF and ML phases of Pre women. During Nal infusion, LH pulse frequency increased in the ML compared to the LF phase of the cycle and, in oophorectomized women, was greater after CE and CE with MPA treatment compared to pretreatment values (P less than 0.05). LH amplitudes during Nal infusion were highest in the ML phase and after CE and MPA treatment in oophorectomized women, and these LH amplitudes were similar. No correlation was found between peripheral plasma beta-EP and Nal-evoked LH responses. No differences were evident in plasma beta-EP levels between Pre and oophorectomized women. In conclusion, 1) endogenous opioid activity is low in oophorectomized women; 2) treatment with estrogen increases opioid activity, and the addition of a progestin increases this activity further; and 3) these data support the contention that sex steroids exert a profound influence on endogenous opioid activity. PMID- 2981086 TI - Oxytocin does not influence adrenocorticotropin secretion in man. AB - Recent studies suggest that oxytocin inhibits ACTH secretion in normal men, whereas previous animal studies indicate that oxytocin is a weak secretagogue of ACTH. We studied the effect of oxytocin on plasma ACTH and cortisol in normal men under three conditions. Using four different doses of oxytocin (30, 60, 90, and 120 mU/kg), given by iv bolus injection, we found no difference in the effect of oxytocin on morning ACTH and cortisol levels compared with the effect of a placebo. A prolonged infusion of oxytocin (90 mU/kg . h) given from 0300-0600 h did not suppress basal ACTH or cortisol levels or delay or blunt their normal circadian rise. In addition, pretreatment with oxytocin (50 mU/kg, iv bolus dose, followed by an infusion of 50 mU/kg . h) did not alter the ACTH and cortisol responses to vasopressin. We conclude that oxytocin is not a major regulator of pituitary-adrenal activity in normal men. PMID- 2981087 TI - Does 1,25-dihydroxyvitamin D participate in the regulation of hormone release from endocrine glands? AB - The presence of receptors for 1,25-dihydroxyvitamin D3 in the pituitary, pancreas, testis, and ovary has raised the question of a possible direct role for 1,25-dihydroxyvitamin D (1,25(OH)2D) in the regulation of hormone synthesis and secretion. To evaluate this problem, six children with the syndrome of resistance to 1,25(OH)2D with rickets and alopecia underwent dynamic tests of insulin, TSH, PRL, GH, and testosterone secretion. Oral glucose loading resulted in normal glucose curves, subnormal peak insulin responses of 12-20 microU/ml in three hypocalcemic patients, and normal peak serum insulin values of 30-40 microU/ml in two normocalcemic patients. Basal serum, TSH, PRL, T4, and T3 concentrations were normal in all patients. Peak serum TSH values after TRH were 11-17 and 16-32 microU/ml in the hypo- and normocalcemic patients, respectively. The PRL response to TRH stimulation in either hypocalcemic or normocalcemic patients was normal [mean 26.2 +/- 5.1 (SD) ng/ml]. Peak serum GH levels were greater than 8 ng/ml in all five patients studied after one or more of the various stimuli. Serum testosterone concentrations after hCG stimulation were normal in the three patients studied (4.1-8.0 ng/ml). Thus, in children with resistance to 1,25(OH)2D, we could find no significant abnormalities in hormone secretion from the pituitary, pancreas, and testis apart from those presumably due to the hypocalcemia itself. PMID- 2981088 TI - Effects of enalapril, a new angiotensin-converting enzyme inhibitor, in a controlled trial in heart failure. AB - Angiotensin-converting enzyme inhibitors are effective vasodilators in the treatment of congestive heart failure. Enalapril, a new angiotensin-converting enzyme inhibitor, or placebo, in addition to digoxin and diuretic drugs, were given to 17 patients with chronic congestive heart failure who were followed up for 12 weeks. In random double-blind fashion, nine patients received enalapril and eight received placebo. Cardiac dimensions and function improved slightly but insignificantly in both groups. Treadmill exercise duration increased from a mean value (+/- standard deviation) of 9.1 +/- 3.2 to 12.0 +/- 3.5 minutes during enalapril administration (p less than 0.025) and was unchanged during placebo administration (10.1 +/- 3.7 versus 11.1 +/- 5.2 minutes). Maximal oxygen consumption also increased during enalapril therapy (15.8 +/- 3.4 to 18.4 +/- 4.4 ml/min per kg, p less than 0.05) and remained unchanged during placebo treatment (16.0 +/- 6.4 versus 17.0 +/- 4.6 ml/min per kg). Clinical functional class (Yale scale) improved 3.1 +/- 1.9 points (p less than 0.01) during enalapril treatment but not during placebo treatment (0.8 +/- 3.5 points, no significant difference). No significant side effects were observed. Thus, enalapril appears to be a clinically effective and useful new angiotensin-converting enzyme inhibitor for the management of chronic congestive heart failure. PMID- 2981089 TI - B cell activation. VI. Effects of exogenous diglyceride and modulators of phospholipid metabolism suggest a central role for diacylglycerol generation in transmembrane signaling by mIg. AB - Previous evidence indicates that in vitro activators of protein kinase C, such as phorbol myristate acetate (PMA), are able to induce early activation events in murine B cells, including membrane depolarization and increased I-A antigen expression. These same events are induced by specific antigen and anti-receptor antibody. This evidence suggests that protein kinase C activation may be an important intermediary event in mIg-mediated transmembrane signaling. Previously, investigators have suggested that protein kinase C activation is regulated by a novel second messenger, diacylglycerol (DG), and DG is generated by phosphatidylinositol (PI) hydrolysis after receptor-ligand interaction in many systems. In view of this concept, we examined the effects of nonspecific activators and inhibitors of DG production and DG itself on membrane potential and levels of I-A antigen expression in murine B cells. Our results indicate that exposure to DG, or induction of DG production by treatment of B cells with exogenous phospholipase C, results in depolarization and increased I-A antigen expression similar to that induced by anti-receptor antibody and specific antigen. Furthermore, we demonstrate that depolarization and increased I-A expression induced by anti-receptor antibody is blocked under conditions in which DG production is inhibited. As expected, based on its direct activation of protein kinase C, PMA stimulation is unaffected by this inhibition. These results support our earlier hypothesis that occupancy of antigen receptors on B cells is linked to subsequent activation events by PI hydrolysis, DG generation, and protein kinase C activation. PMID- 2981090 TI - Defective IFN-gamma production in the human neonate. II. Role of increased sensitivity to the suppressive effects of prostaglandin E. AB - Analysis of endogenous production and effects of exogenous addition of interleukin 2 (IL 2), leukotrienes (LT), and prostaglandin E (PGE) has been used to investigate the dysregulation responsible for impaired PHA-induced IFN-gamma secretion by cord blood leukocytes (CBL). The addition of LT or IL 2 could not reverse the IFN defect of CBL. The production of these two mediators was found to be normal in CBL cultures. CBL and control leukocytes from adult donors produced comparable amounts of PGE2. In contrast, sensitivity to the suppressive effects of PGE2 on IFN-gamma secretion was much higher with CBL than with control leukocytes. Treatment with indomethacin reversed the IFN-gamma defect with most CBL tested, and the addition of physiologic amounts of PGE2 to indomethacin treated cultures resulted in a profound impairment of IFN-gamma production similar to that of untreated CBL cultures. Preincubation of CBL for 24 hr before PHA stimulation resulted in restoration of a normal sensitivity to exogenous PGE2, in parallel with correction of the IFN-gamma defect. Our observations suggest that the impairment of IFN-gamma secretion in neonates is not due to deficient amplification circuits, but is the consequence of an exaggerated cellular sensitivity to the suppressive effects of PGE produced endogenously in normal amounts. PMID- 2981091 TI - Human immune responses to synthetic peptides from the Epstein-Barr nuclear antigen. AB - Humans infected with Epstein-Barr virus (EBV), the causative agent of infectious mononucleosis, develop antibodies against a nuclear antigen (EBNA) that is present in virally transformed B lymphocytes. The EBNA protein contains a unique glycine-alanine repeating sequence. We have synthesized peptides corresponding to various regions of the EBNA molecule within and near this sequence. Rabbit antibodies against the peptides within the sequence reacted directly with the EBNA protein, as detected by Western blotting. The sera of individuals with antibodies against Epstein-Barr virus contained abundant antibodies also reactive with one or several of the synthetic peptides within the sequence. Moreover, human antibodies against these simple peptides were induced specifically early in the course of infectious mononucleosis. When compared with normal controls, antibody levels to the glycine-alanine peptides were significantly higher in patients with rheumatoid arthritis and progressive systemic sclerosis, but not in patients with two other autoimmune diseases. These results document that i) antibodies against the peptides detect the EBNA protein, ii) humans infected with EBV produce high titers of antibodies reactive with these synthetic antigens, and iii) antibody titers against the peptides are abnormally elevated in certain autoimmune diseases. PMID- 2981092 TI - Biochemical models of gamma-interferon action: altered expression of transferrin receptors on murine peritoneal macrophages after treatment in vitro with PMA or A23187. AB - The number of transferrin receptors in thioglycollate-elicited murine peritoneal macrophages is markedly depressed after exposure to murine gamma-interferon (IFN gamma) in vitro. This change has been used as a model system to study the molecular and cellular mechanisms of IFN gamma signal transduction. We observed that the downshift of the transferrin receptor could be mimicked by exposure to the calcium ionophore (A23187) or the potent tumor promoter, phorbol 12-myristate 13-acetate (PMA). Saturation binding studies on thioglycollate (TG)-elicited peritoneal macrophages after exposure to A23187 or PMA showed the reduced expression of transferrin binding activity attributable to a decrease in the total number of cellular transferrin receptors and not an alteration in receptor ligand affinity, in agreement with previous results obtained after exposure to IFN gamma. The loss of transferrin receptors in response to A23187 or PMA was dose dependent, and the kinetics of the change were identical to those observed with IFN gamma treatment. Phorbol 12,13-dibutyrate or 4-beta-phorbol 12,13 didecanoate, both biologically active phorbol esters, also induced reduced expression of transferrin receptors, whereas nonesterified phorbol or 4-alpha phorbol 12,13-didecanoate, an inactive phorbol ester, had no effect on transferrin receptor expression. Finally, PMA and A23187, when used together, acted cooperatively to modulate transferrin receptor expression when both agents were present at subthreshold concentrations. These results, taken together, suggest that elevation of intracellular Ca++ levels and/or stimulation of protein kinase C are involved in the response of macrophages to IFN gamma. PMID- 2981093 TI - Purification and distribution of a novel macrophage-specific calmodulin-binding glycoprotein. AB - The murine macrophage-like cell line J774.16 and peritoneal exudate cells elicited with thioglycollate or starch contain a major calmodulin (CaM)-binding protein (CaMBP) which is absent in trifluoperazine-resistant variants of J774, resident peritoneal macrophages and peritoneal macrophages elicited with concanavalin A, lipopolysaccharide, proteose peptone, or Bacillus Calmette Guerin. Resident murine peritoneal cells maintained in tissue culture for 3 days begin to accumulate this protein, as do human peripheral blood monocytes after 7 days of culture. A specific competitive displacement radioimmunoassay was developed with the use of a rabbit antiserum raised to the partially purified CaM binding protein and [125I]CaM covalently cross-linked to the principal CaM binding protein in the preparation. The radioimmunoassay confirmed the unique cellular distribution of this protein, suggesting that it may be a marker for certain stages of macrophage differentiation. Monoclonal antibodies were prepared, and one of these was used to further purify the protein by immunoaffinity chromatography. A protein of Mr 50,000 to 60,000 was isolated. The protein could be selectively adsorbed to wheat germ agglutinin agarose and subsequently eluted with N-acetyl glucosamine. This property, plus the sensitivity of the protein to endoglycosidase F, led to the conclusion that it is a glycoprotein. The cellular distribution, subcellular localization, and evidence for glycosylation suggest that this protein may be a macrophage-specific receptor with a high affinity for Ca2+-CaM. PMID- 2981094 TI - Natural killer cells vs cytotoxic T cells in the peripheral blood of virus infected mice. AB - The cell-mediated immune response of mice against various enveloped RNA and DNA viruses expressed by immune lymphocytes from the spleen and the peripheral blood (PBL) were compared. PBL from mice of various strains infected with vaccinia virus, vesicular stomatitis virus (VSV), or lymphocytic choriomeningitis virus (LCMV) were tested on histocompatible or incompatible target cells infected with the homologous virus. PBL from immune mice showed clear H-2 restriction, but additionally, they expressed high natural killing (NK) activity on YAC-1 cells. The high NK-cytolytic activity of PBL on YAC-1 differed significantly from that expressed by splenic lymphocytes. In both lymphocyte populations lysis was detected as early as 1 day after infection; NK activity decreased in the spleen after day 4 post infection, whereas that of PBL persisted at high levels for up to 10 days after infection. Treatment of mice with anti-asialo GM1 in vivo abrogated NK activity in PBL effector cells tested in vitro. These results may explain some of the difficulties to observe MHC-restricted cytotoxic T cells in PBL from humans or primates during primary infections with virus. PMID- 2981095 TI - B cell subsets in spleens of BALB/c mice: identification and isolation of MMTV expressing and MMTV-responding subpopulations. AB - The DNA of BALB/c mice contains two genomic- and one subgenomic-size MMTV proviruses that appear to be preferentially expressed in their spleen cells, although intact MMTV virions cannot be detected in the tissues of these mice. This retrovirus antigen expression is restricted to a subpopulation of B lymphocytes, as determined by double label immunofluorescence studies. Nylon adherent, SIg-positive spleen lymphocytes from BALB/c mice are capable of being stimulated by purified MMTV in lymphocyte transformation assays. Two possibilities were explored: the MMTV-positive cells are the responders to MMTV in the blastogenesis assay, or there exists two B lymphocyte subsets, one expressing the MMTV antigen(s) and the other responding to the virus. Depletion of MMTV-positive, nylon-adherent cells with anti-MMTV and complement resulted in no significant change in the blastogenesis to MMTV, indicating that the MMTV negative lymphocytes are the responders in this reaction. These results were confirmed by positive selection experiments by using a fluorescence-activated cell sorter. Two populations of nylon-adherent cells, on the basis of the presence or absence of MMTV antigen in their surfaces, were obtained by a two-way sorting procedure and were used in lymphocyte transformation assays. MMTV expressing lymphocytes were found to be nonresponsive to MMTV antigens, although high levels of [3H]thymidine incorporation were observed in the MMTV-negative, nylon-adherent cell cultures exposed to MMTV. These data indicate that in normal BALB/c mice, expression of endogeneous retrovirus genetic information is restricted to a nylon-adherent spleen cell subset that is different from the one responding in blastogenesis to the viral antigens. PMID- 2981096 TI - Comparison of MHC genes among distantly related members of the genus Mus. AB - The genomic content of class I and class II MHC DNA sequences in a variety of wild mice has been analyzed. The panel of mice includes members of three subgenera of the genus Mus. By genomic hybridization with the use of a variety of DNA probes, both class I and class II DNA sequences appear to be conserved in all of the species examined. However, the number of class I DNA sequences differs among the species. Furthermore, this variation appears to result from differential increases within subsets of class I genes. These data suggest that the class I multigene family is dynamic and changing over short periods of evolutionary time. In contrast, none of the class II genes appears to vary in copy number. More extensive polymorphism was noted amongst the class II beta genes than the alpha genes. Interestingly, the genomic sequence corresponding to E beta 2 is highly conserved, leading to the prediction that it is a genetically functional sequence. PMID- 2981097 TI - Solubilization and detergent effects on interactions of some drugs and insecticides with the t-butylbicyclophosphorothionate binding site within the gamma-aminobutyric acid receptor-ionophore complex. AB - Specific binding of [35S]t-butylbicyclophosphorothionate (TBPS) to rat brain membranes (RBM) is enhanced nine-fold by EDTA/water dialysis and 1.3- to 4.2-fold by 50 nM ketosteroid R 5135, or 5 mM 3-[(3-cholamidopropyl)dimethylammonio]-1 propanesulfonate (CHAPS) or related piperazine-N-alkanesulfonate buffers, or extensive washing with NaCl/Na phosphate or Na phosphate/citrate solution. About one-fifth of the [35S]TBPS binding capacity appears in the soluble fraction whereas the rest remains in particulate form on treatment of the EDTA/water dialyzed RBM with 20 mM CHAPS. Similar KD values (64-86 nM) are obtained for the original EDTA/water-dialyzed membranes and the CHAPS-treated and/or -solubilized preparations. The Bmax of the EDTA-treated RBM is reduced five-fold on solubilization with CHAPS. The potency for displacement of [35S]TBPS changes in the presence of CHAPS or on CHAPS solubilization: gamma-aminobutyric acid (GABA) and muscimol inhibit specific [35S]TBPS binding more strongly in the absence than in the presence of CHAPS: TBPS, picrotoxinin, and photoheptachlor epoxide are almost equally active with RBM, RBM + CHAPS, and RBM solubilized with CHAPS. Levels of (1R, alpha S)-cis-cypermethrin and dimethylbutylbarbiturate which are inhibitory with RBM are moderately stimulatory after TBPS receptor solubilization. Thus CHAPS defines three regions of the GABA receptor-ionophore complex, i.e., the GABA and benzodiazepine receptors, the TBPS/picrotoxinin/polychlorocycloalkane receptor(s), and the sites at which the alpha-cyano pyrethroid and the barbiturate interact with TBPS binding. PMID- 2981098 TI - Purification of phosphatidylinositol synthetase from rat brain by CDP diacylglycerol affinity chromatography and properties of the purified enzyme. AB - A purification procedure for rat brain phosphatidylinositol synthetase (PI synthetase; CDP-1,2-diacyl-sn-glycerol:myo-inositol 3-phosphatidyltransferase; EC 2.7.8.11) is described. The enzyme was purified 200-250-fold from the homogenate by solubilization with Triton X-100 from microsomal membranes and affinity chromatography on CDP-diacylglycerol-Sepharose. Elution of enzyme activity required the presence of Triton X-100, CDP-diacylglycerol, and either phosphatidylcholine or asolectin. The product that was obtained in 5-10% yield from whole brain and in 70% yield from the microsomal fraction contained three protein bands as determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The final preparation contained levels of CDP diacylglycerol hydrolase and CDP-diacylglycerol: sn-glycero-3-phosphate 3 phosphatidyltransferase activities that were less than 1% of PI synthetase activity. The purified enzyme displayed a pH optimum of 8.5-9.0, required either Mg2+ or Mn2+ and exhibited a Km of 4.6 mM for myo-inositol. PMID- 2981099 TI - Effects of guanyl nucleotides on [3H]flunitrazepam binding to rat hippocampal synaptic membranes: equilibrium binding and dissociation kinetics. AB - The effects of guanyl nucleotides on the binding of [3H]flunitrazepam to rat hippocampal synaptic membranes were studied. In equilibrium binding studies, gamma-amino-n-butyric acid (GABA) increased and GTP decreased the binding affinity of [3H]flunitrazepam; GTP also caused a decrease in binding capacity. The effect, however, is variable. In studies of the dissociation kinetics of [3H]flunitrazepam using diazepam and the antagonist Ro 15-1788 as the displacers, there was evidence of two dissociation rate constants. GTP increased both the fast- and slow-dissociation rate constants and increased the ratio of the slow dissociation binding state. The effect of GTP was mimicked by its nonhydrolyzable analogue 5'-guanylylimidodiphosphate but not by ATP and occurred when diazepam, but not when Ro 15-1788, was used as the displacer. GABA antagonized the effect of GTP on the dissociation of [3H]flunitrazepam. The nature of the benzodiazepine receptor, its actions, and the possible role of cyclic AMP as a second messenger are discussed. PMID- 2981100 TI - Kinetics of the mechanism of action of monoamine oxidase in the regulation of Na+,K+-ATPase activity in rat brain. AB - Kinetic studies on the action of monoamine oxidase (MAO) in the regulation of Na+,K+-ATPase were performed using 3-methoxy-4-hydroxybenzaldehyde (MHB), which is an analogue of 3-methoxy-4-hydroxy-phenylacetylaldehyde (product of MAO catalysed reaction with dopamine as substrate). It was observed that at 2.6 microM MHB, the activation of Na+,K+-ATPase may be the result of the removal of the inhibitory Ca2+, thereby increasing the Vmax. Double-reciprocal plots of Pi versus MHB showed that Ca2+ counteracted the effect of the aldehyde not by changing the Km, but be decreasing the Vmax of the Na+,K+-ATPase stimulation. The removal of 3',5'-cyclic AMP-dependent protein kinase from the microsomes by sodium dodecyl sulphate treatment abolished the activation and/or inhibition of the Na+,K+-ATPase by aldehyde; it can therefore be inferred that 3',5'-cyclic AMP dependent protein kinase is involved in the regulation of Na+,K+-ATPase. PMID- 2981101 TI - Allosteric modulation of flunitrazepam binding to rat brain benzodiazepine receptors by methyl beta-carboline-3-carboxylate. AB - The inhibition of flunitrazepam (FNP) binding to rat brain benzodiazepine (BZ) receptors by methyl beta-carboline-3-carboxylate (MCC) was studied. Biphasic dissociation was observed for [3H]FNP and [3H]MCC in cerebral cortex, cerebellum, and hippocampus, although the dissociation of [3H]MCC was much faster. The dissociation rate of [3H]FNP was increased by MCC in the cerebellum, but was not altered in cerebral cortex or hippocampus. [3H]FNP binding stimulated by gamma aminobutyric acid was enhanced in the presence of MCC in all three regions examined. These results indicate that MCC exerts these effects by interacting with allosteric sites that are different from the FNP recognition sites on the BZ receptors. PMID- 2981102 TI - Relationship between hemispheric cerebral blood flow, central conduction time, and clinical grade in aneurysmal subarachnoid hemorrhage. AB - The relationship between central conduction time (CCT) and hemispheric cerebral blood flow (CBF) has been examined in 20 patients presenting with subarachnoid hemorrhage. A total of 63 combined CCT/CBF recordings were performed at various times throughout the hospital course of these patients, and the findings were correlated to clinical status. The initial-slope index of the CBF (CBF isi) was found to correlate well with clinical grade, and a gradation in flow was noted between the different neurological grades. Patients in Grades I and II (Hunt and Hess classification) had the highest flows (mean CBF isi = 47.2 +/- 8.1); Grade III patients had intermediate flows (mean CBF isi = 39.6 +/- 7.8); and Grade IV patients had the lowest flows (mean CBF isi = 32.0 +/- 6.4). While CCT tended to become increasingly prolonged with worsening grade, a significant difference could not be demonstrated between Grade I, II, and III patients. Only when Grade IV status was reached was the CCT significantly prolonged. When CBF isi and CCT were examined, a threshold relationship was noted between CBF isi and CCT prolongation. At flow values above 30, little change was noted in CCT, and CCT remained in the normal range. However, at flow values below 30, CCT became increasingly prolonged as blood flow diminished. The degree of CCT prolongation appeared to be directly proportional to the degree of blood flow diminution at flows below threshold. PMID- 2981103 TI - Results of neurosurgical treatment by a transsphenoidal approach in 10 patients with Nelson's syndrome. AB - Ten patients with Nelson's syndrome, nine women and one man, aged 22 to 61 years, were treated neurosurgically by a transsphenoidal approach. In four patients, microadenomas were found, ranging in diameter from 4 to 10 mm. Microscopically, mixed adenoma was diagnosed in six cases, basophilic adenoma in three patients, and chromophobe adenoma in one patient. The presence of argyrophilic nerve fibers in the adenoma tissue was noted in one case. The time of observation after transsphenoidal surgery ranged from 6 months to 10 years. Clinical remission was achieved in eight patients; in two of them, radiation therapy was used to complement surgical treatment. In two patients, recurrence of the pituitary neoplasm was observed; anaplasia was revealed at the second operation in one of them. Radiation therapy might be a useful adjunct to neurosurgery in Nelson's syndrome, especially in patients with anaplastic adenoma. PMID- 2981105 TI - A case of nonfunctioning paraganglioma arising from the retroperitoneum: angiographic and scintigraphic features. AB - We report a case of a nonfunctioning nonchromaffin paraganglioma arising from organ of Zuckerkandl. Angiography manifested a cartwheel-shaped image in the arterial phase and a characteristic tortoise shell-shaped lobular structure with homogeneous opacification in the venous phase. Just after administration of 99mtechnetium-E-HIDA scintigraphy disclosed a globular hot density below the left kidney. These findings may represent characteristic signs of paragangliomas. PMID- 2981104 TI - Effects of vasoactive intestinal polypeptide on isolated urethral and urinary bladder smooth muscle from rabbit and man. AB - Vasoactive intestinal polypeptide concentration-dependently inhibited the contractant responses of isolated preparations of the female rabbit bladder and urethra induced by electrical field stimulation and exogenous application of acetylcholine (bladder) and noradrenaline (urethra). The inhibition of alpha adrenoceptor and muscarinic cholinoceptor-mediated activity in the urethra and bladder amounted to 50 to 90 per cent of induced contractions. The nonadrenergic noncholinergic contraction induced by electrical field stimulation in the urethra was reduced slightly, whereas corresponding response in the bladder was more sensitive. The maximum inhibition of both the electrically induced responses and contractions induced by exogenous noradrenaline and acetylcholine was of comparable size in the urethra and the bladder. The effects of vasoactive intestinal polypeptide seemed to be exerted postjunctionally since no significant influence of the peptide was seen on the release of 3H-noradrenaline from adrenergic nerve endings in the urethra. The effects of vasoactive intestinal polypeptide in human urethral and bladder preparations were less consistent. The noradrenaline-induced contraction in urethral preparations was inhibited by 29 +/ 9 per cent (no. = 22). The effects on electrically induced contractions in the urethra, and on responses to acetylcholine and electrical field stimulation in the bladder, were small and inconsistent. It is concluded that vasoactive intestinal polypeptide may be of importance for regulation of lower urinary tract smooth muscle activity in the rabbit. It cannot be excluded that the peptide has a modulatory role in neurotransmission in human urethral muscle. However, the present results do not support the view of vasoactive intestinal polypeptide as an inhibitor of contraction in human detrusor. PMID- 2981106 TI - Risk of nosocomial infection with human T-cell lymphotropic virus III (HTLV-III). AB - Infection with human T-cell lymphotropic virus III (HTLV-III) is closely linked to the acquired immunodeficiency syndrome (AIDS). We evaluated the risk of nosocomial infection with HTLV-III by testing for antibodies to HTLV-III among hospital employees, including victims of needle-stick exposure, endoscopists, pathologists, and laboratory workers. Assays for antibody against the virus were performed by enzyme-linked immunosorbent assay and electrophoretic (Western blot) techniques. Although all 22 of our patients with AIDS and 6 of 7 with AIDS related complex were found to have antibodies to HTLV-III when both assays were employed, none of the 85 employees with nosocomial exposure to specimens from patients with AIDS were positive for HTLV-III antibody. These studies must be regarded as preliminary, but they suggest that when current hospital isolation procedures are employed, the risk of nosocomial transmission of HTLV-III is low. PMID- 2981108 TI - Preoperative lateralization of ACTH-secreting pituitary microadenomas by bilateral and simultaneous inferior petrosal venous sinus sampling. PMID- 2981107 TI - Acromegaly due to secretion of growth hormone by an ectopic pancreatic islet-cell tumor. PMID- 2981109 TI - Non-Hodgkin's lymphoma and HTLV-1 antibodies in a gorilla. PMID- 2981110 TI - Post-traumatic gastric stenosis due to perigastric adhesions. AB - A case of perigastric adhesions that were caused by a gunshot wound and that simulated scirrhous carcinoma is described. PMID- 2981111 TI - Hepatocellular carcinoma and metastatic cancer detected by iodized oil. AB - An oily contrast medium was injected into the hepatic artery in rabbits with implanted VX2 carcinoma in the liver. The contrast medium was initially detected in all of the branches of the injected artery, but 3 and 7 days after injection it was found only in the tumor tissue on plain radiographs. Taking advantage of this phenomenon, an oily contrast medium was injected into the hepatic artery in 5 patients with liver tumors. A plain radiograph of the abdomen after the injection presented a clearer picture of the tumor and detected smaller tumors, compared with a conventional selective celiac angiogram in 5 patients. It is concluded that an injection of oily contrast medium into a hepatic artery is a useful technique to detect liver tumor. PMID- 2981113 TI - Correlation of scintigraphy and angiography for ventricular volume determinations using realistic cardiac phantoms: the unimportance of self-attenuation. AB - The volumes of the left ventricle (LV) and right ventricle (RV) were determined by scintigraphy, radiography, and weight, using hollow, morphologically accurate cardiac phantoms that ranged from 99-415 ml (RV) and 72-364 ml (LV). Self attenuation within these simulated ventricles appeared negligible in view of the nearly linear correlation between scintigraphic and weight determined volumes. Scintigraphy compared favorably with radiography for estimating clinically encountered volumes of the RV and LV. PMID- 2981112 TI - Hepatocellular carcinoma detected by iodized oil. AB - This study assesses the diagnostic value of Lipiodol (iodized oil) and computed tomography (CT) in detecting hepatocellular carcinoma (HCC). Twenty-four patients who were suspected of having HCC received injections of a small amount of Lipiodol, along with an antitumor agent, in the hepatic artery following routine celiac angiography. CT scans obtained 7-10 days after Lipiodol administration demonstrated HCC in distinct contrast to the surrounding noncancerous parenchyma. In particular, the CT-Lipiodol procedure disclosed many small HCC lesions that were not shown by celiac angiography, scintigraphy, CT with and without contrast medium enhancement, and ultrasonography. Although this procedure may miss very small or highly fibrotic lesions, it is recommended for patients suspected of having HCC and for patients for whom hepatic resection is being considered. PMID- 2981114 TI - Hepatocellular carcinoma detected by iodized oil. Use of anticancer agents. AB - Transcatheter arterial embolization (TAE) was performed in 97 patients with hepatocellular carcinoma. Ethiodol (iodized oil) containing an anticancer drug was infused via the hepatic artery followed by Gelfoam particles. The Ethiodol emulsion was selectively retained in the tumor vessels and also remained in the small daughter nodules that could not be detected by angiography or computed tomography (CT) prior to TAE. Ethiodol remained in the tumor for more than 1 year. Following TAE, main tumors and small daughter nodules appeared as areas of markedly high density on CT. In most patients there was a reduction in the tumor size following TAE, and serum alpha-fetoprotein levels were reduced in all patients whose initial levels had exceeded 400 ng/ml. This method is considered to be effective not only for treatment of hepatic tumor but also useful for evaluation of post-TAE changes in the tumor and diagnosis of small daughter nodules, due to the long-term accumulation of Ethiodol in tumor vessels. PMID- 2981115 TI - Integrated imaging of hepatic tumors in childhood. Part I: Malignant lesions (primary and metastatic). AB - Both the prognosis and treatment of hepatic tumors in children depend upon the histological diagnosis and the extent of disease. Recent advances in imaging techniques permit characterization of specific tumors and differentiation from other intrahepatic processes. An integrated imaging protocol involving a combination of ultrasound, computed tomography, and scintigraphy often provides a high degree of accuracy. Patterns derived from 40 cases of hepatoblastoma, hepatocellular carcinoma, rhabdomyosarcoma, monotypic small-cell sarcoma, and metastatic tumors are discussed and an algorithm for evaluation of hepatic tumors in children is presented. PMID- 2981116 TI - A mammalian host-vector system that regulates expression and amplification of transfected genes by temperature induction. AB - SV40-transformed simian cells that permit temperature-dependent regulation of vector DNA replication were isolated and characterized. These cell lines (ts COS cells) produce high levels of thermolabile large T antigen under the transcriptional control of the Rous sarcoma virus long terminal repeat. The ts COS cell lines can complement SV40 A gene mutants and support replication of SV40 origin containing vectors at 33 degrees C but not at 40 degrees C. It should now be possible to regulate the copy number of transfected plasmid DNA's and also maintain selectable vector sequences either as integrated DNA or as autonomously replicating episomes by modulating T antigen activity in ts COS cells. PMID- 2981117 TI - Enzyme regulation in a trypanosomatid: effect of purine starvation on levels of 3'-nucleotidase activity. AB - Crithidia fasciculata, a nonpathogenic relative of the leishmanial and trypanosomal pathogens of humans and animals, showed a 3'-ribonucleotidase activity similar to that in Leishmania donovani. The level of 3'-nucleotidase activity in Crithidia was regulated by the availability of purines in the culture medium. Specifically, organisms obtained from culture medium depleted of purines contained elevated levels of enzyme activity compared to those grown in complete medium. The 3'-nucleotidase, located at the cell surface, may serve as a first step in purine salvage for these protozoa, which are unable to synthesize the purine ring de novo. PMID- 2981118 TI - Cytomegalovirus infection in patients undergoing noncardiac surgical procedures. AB - A search for records for CMV infection following transfusion related to noncardiac and nontransplant operations and trauma disclosed nine instances with CMV seroconversion. One patient had received only a single unit of blood. The most prominent feature was fever with a characteristic spiking plateau pattern, which began approximately three weeks after blood transfusion. Splenomegaly and atypical lymphocytosis were less common. The results of hepatic function tests showed slight abnormalities. Associated splenectomy did not result in a more severe manifestation of the CMV infection. The late postoperative fever in these patients led to an extensive and costly investigation before determination of antibody titers to CMV and confirmation of seroconversion. When faced with the constellation of symptoms, including a delayed (two to three weeks) spiking plateau postoperative fever, abnormal results of hepatic function test and lymphocytosis in patients having received blood transfusion, the clinician must give serious consideration to the possibility of CMV infection. PMID- 2981119 TI - Malignant chemodectoma of the carotid body causing spinal cord compression. AB - We present a case of a 19-year-old woman with a malignant chemodectoma of the carotid body invading the cervical spine. The interest in the case is due to the rarity both of the lesion and of the spine invasion causing a medullary compression. PMID- 2981120 TI - Parasellar metastases: four autopsied cases. AB - Four patients with histologically confirmed parasellar metastases are reported. The main symptoms and signs were persistent right facial pain followed by diplopia (patient 1), headache and minimal right abducens palsy (patient 2), acute, total left ophthalmoplegia (patient 3), and acute, total bilateral ophthalmoplegia (patient 4). Positive radiologic evidence was present only in patient 1: there was bony erosion of the petrous apex and computed tomography scan showed an enhanced parasellar mass. This patient underwent partial surgical removal of the tumor. Patient 3 was treated with irradiation. All patients died within 14 weeks of the onset of the initial symptoms and all were autopsied. Their primary lesions were hepatoma, stomach cancer, lung cancer, and mesenteric liposarcoma. PMID- 2981121 TI - Parkinsonism as a manifestation of brain tumor. AB - Neoplasm is an uncommon cause of the parkinsonian syndrome. We are presenting three cases of intracranial tumors whose major manifestations included parkinsonism. Surgical treatment resulted in control of the extrapyramidal symptoms in two of our patients. Because no curative treatment is available for the vast majority of parkinsonian patients, early detection of an underlying neoplasm may offer a most rewarding outcome. Unilaterality of the symptoms, overwhelming dementia, or suspicion of a mass should lead to the performance of computed tomography. PMID- 2981122 TI - Glioblastoma multiforme. PMID- 2981123 TI - Gallbladder infarction after hepatic artery embolization. AB - Nineteen patients with hepatocellular carcinoma had transcatheter hepatic arterial embolization and subsequent hepatic resection with cholecystectomy. During the angiographic procedure, inadvertent embolization of the cystic artery occurred in 10 patients (53%). Subsequent histologic study of the gallbladder demonstrated necrotizing ulcerative cholecystitis in nine of the 10. All nine patients complained of right-upper-quadrant pain of varying degree after hepatic embolization, but none required special treatment or urgent laparotomy. PMID- 2981124 TI - CT of perineural tumor extension: pterygopalatine fossa. AB - Tumors of the oral cavity and paranasal sinuses can spread along nerves to areas apparently removed from the primary tumor. In tumors of the palate, sinuses, and face, this "perineural" spread usually involves the maxillary division of the trigeminal nerve. The pterygopalatine fossa is a pathway of the maxillary nerve and becomes a key landmark in the detection of neural metastasis by computed tomography (CT). Obliteration of the fat in the fossa suggests pathology. Case material illustrating neural extension is presented and the CT findings are described. PMID- 2981125 TI - CT of the gluteal region. PMID- 2981126 TI - Hepatitis A antibodies: prevalence and persistence in a group of Mexican children. AB - The prevalence of antibodies against hepatitis A virus (anti-HAV) was evaluated in sera from 275 asymptomatic infants and children under the age of five years and in 46 maternal-cord serum pairs, followed by sera obtained thereafter each month from the first to the fifth month of life in 29, 17, 9, 9 and 7 infants (a total of 163 serum samples). Anti-HAV was found in all cord blood sera, but its prevalence declined progressively to 0% among 19 infants aged eight months. Four (25%) of 16 children aged one year had positive anti-HAV and the antibody prevalence was 89% in children five years old. Recent hepatitis A infection shown by circulating anti-HAV immunoglobulin M was detected in five infants under the age of two months and with frequencies ranging from 19-67% in one- and five-year old children. It was concluded that in children living in Mexico City, hepatitis A infections occur at an earlier age and at higher rates than in children in other cities, especially those in developed countries. PMID- 2981127 TI - Effect of protein concentration on kinetic constants (Kd and Bmax) of 3H imipramine binding in blood platelets. AB - 3H-imipramine binding was studied in the blood platelets of 80 normal controls and 50 depressed (psychotic and nonpsychotic) patients. Protein concentration in the incubation mixture, and Kd and Bmax values were correlated in normal controls and depressed patients. A small but significant correlation between protein in the incubation mixture and Bmax was observed in normal controls and depressed patients. There was also a significant correlation between protein and Kd of imipramine binding in the blood platelets of unipolar psychotic depressed patients. However, analysis of covariance to remove the effect of protein on Kd and Bmax did not change the basic finding of decreased Bmax in the blood platelets of psychotic depressed patients. PMID- 2981128 TI - Lithium incorporation in the fibroblasts of manic-depressives. AB - Lithium incorporation in cultured human skin fibroblasts was measured in a group of 10 manic-depressives and 10 controls. This system was believed to eliminate many of the sources of variability to which measurement of the lithium ratio in erythrocytes (RBCs) is subject. A fibroblast lithium ratio calculated from 1-hr lithium incorporation studies correlated highly with an in vitro RBC lithium ratio in medication-free controls. Manic-depressives and controls did not differ in lithium incorporation or fibroblast lithium ratio, leading to the conclusion that the lithium ratio is probably not a good measure for differentiating the two populations. PMID- 2981129 TI - Nontolerance to the opioid antagonism of naltrexone. AB - Controlled opiate challenges of naltrexone-pretreated human subjects have established that naltrexone is an effective opioid antagonist. However, these challenges have been conducted after relatively acute dosing with naltrexone, and tolerance to this antagonism after chronic treatment is possible. We therefore administered morphine challenges in a double-blind, placebo-controlled design to nine ex-addicts who had been taking naltrexone for a mean of 9.4 months. None of the ex-addicts experienced euphoria; instead, most of these blockaded ex-addicts had a dysphoric histaminelike response to the intravenous morphine. The only physiological change was a slight increase in heart rate. We conclude that tolerance does not develop to the opioid antagonist properties of naltrexone up to as long as 21 months of treatment. PMID- 2981131 TI - Chemistry of a human monocyte-derived cell line (U937): identification of the angiotensin I-converting activity as leukocyte cathepsin G. AB - Angiotensin-converting enzyme, a dipeptidyl carboxypeptidase, catalyzes the conversion of angiotensin I to the vasoactive peptide angiotensin II. The finding of angiotensin-converting enzyme in dexamethasone-stimulated cultured monocytes and alveolar macrophages prompted the examination of a human monocyte-like cell line (U937) for angiotensin I-converting activity. Conversion of angiotensin I (5 X 10(-5) mol/L) to angiotensin II by U937 cell extracts (10(4) - 4 X 10(6) cells) was detected, and the pH optimum for the reaction was 7.0 to 8.0. The U937 cell angiotensin I-converting activity was purified to homogeneity by carboxymethylcellulose chromatography and trasylol affinity chromatography. The purified protein performed similarly to purified human neutrophil cathepsin G on sodium dodecyl sulfate-gradient polyacrylamide gel electrophoresis (SDS-gradient PAGE), elicited a reaction of complete identity with neutrophil cathepsin G when diffused against anti-cathepsin G antiserum, and had quantitatively similar angiotensin I-converting activity as neutrophil cathepsin G. Neutrophils and U937 cells had 143 and 52 times greater angiotensin I-converting capability than cultured monocytes or peripheral blood mononuclear cells, suggesting the relative importance of mobile cells containing cathepsin G in the local generation of angiotensin II. These data identify the angiotensin I-converting activity of the U937 cell as leukocyte cathepsin G and provide evidence that the U937 cell has neutrophil-like as well as monocyte-like characteristics. PMID- 2981132 TI - Myeloperoxidase and terminal deoxynucleotidyl transferase in leukemic cells. PMID- 2981130 TI - Increased binding of fibrinogen to platelets in diabetes: the role of prostaglandins and thromboxane. AB - Previous studies suggested a role for prostaglandins or thromboxane A2, or both in the exposure of fibrinogen receptors on normal platelets in response to several aggregating agents. Platelets from diabetics are known to be more sensitive to aggregating agents and to produce more prostaglandins and thromboxane than platelets from normal subjects. We compared fibrinogen binding to platelets from diabetic subjects with binding to platelets from normal subjects and determined whether aspirin (which inhibits the formation of prostaglandins and thromboxane) would inhibit the binding of fibrinogen to platelets from diabetic subjects and whether this correlated with its effects on platelet aggregation. We found the following: Aspirin suppressed thromboxane formation and rendered the platelets less sensitive to the induction of aggregation by adenosine diphosphate (ADP) or collagen. The amount of U-46619 [( 15s]-hydroxy-11-alpha, 9-alpha [epoxy-methano]-prosta[5Z,13E]-dienoic acid, a stable analog of prostaglandin endoperoxide/thromboxane A2) necessary to induce aggregation, was similar in normal and diabetic subjects and was unchanged after ingestion of aspirin. Binding of 125I-fibrinogen following stimulation of platelets by ADP or collagen was greater in diabetic (because more binding sites were exposed) than in normal subjects. However, following stimulation by U-46619, binding was similar in diabetic and normal subjects. Aspirin caused a reduction in the exposure of binding sites on both platelets from diabetic and normal subjects, so that (in this respect) platelets from diabetic subjects became more like those from normal subjects. Effects of the monoclonal antibody B59.2, which is specific for the platelet glycoprotein IIb-IIIa complex (the presumed receptor for fibrinogen on the platelet surface) were also studied. The amount of this antibody that bound to platelets was the same for normal and diabetic subjects both before and after aspirin and with or without stimulation by ADP or collagen. In addition, B59.2 inhibited aggregation and fibrinogen binding in both platelets from diabetic and normal subjects. The combined data suggest that the glycoprotein IIb-IIIa complex of platelets from diabetic subjects is similar to that of platelets from normal subjects and that the increased fibrinogen binding and aggregation of platelets from diabetic subjects in response to ADP or collagen is mediated by increased formation of prostaglandin endoperoxide or thromboxane A2, or both. PMID- 2981133 TI - Volume-dependent human blood polymorphonuclear leukocyte heterogeneity demonstrated with counterflow centrifugal elutriation. AB - Human peripheral blood polymorphonuclear leukocytes (PMNs) have recently been recognized as a heterogeneous population of cells. Consideration has not been given to the possibility that size may be an additional physical characteristic demonstrating heterogeneity. Using counterflow centrifugal elutriation, we have demonstrated that PMNs can be isolated into at least six volume-dependent fractions. A positive correlation exists for PMN size and superoxide anion release upon stimulation with f-Met-Leu-Phe or phorbol myristate acetate. Total granule contents were also noted to be greater in larger PMN fractions, with a constant percent of release upon stimulation. The implications of these findings are discussed. PMID- 2981134 TI - Why do people seek treatment by alternative medicine? PMID- 2981136 TI - Laboratory reports of herpesvirus infections in Canada in 1983. PMID- 2981135 TI - Arrest of neuropathy and myopathy in abetalipoproteinemia with high-dose vitamin E therapy. AB - A 16-year-old girl, one of dizygotic twins, presented in 1976 complaining of a 1 year history of a lack of coordination and an inability to run. The results of biochemical tests confirmed the diagnosis of classic abetalipoproteinemia. In addition to the recognized neurologic features of this disorder, she had a reduced evoked motor unit potential and markedly elevated serum levels of muscle enzymes, which suggested myositis. The serum vitamin E level was markedly decreased. Oral therapy with vitamin E, 800 mg daily, was begun, and in 1981 the dosage was increased to 3200 mg daily. Over the 7 years of follow-up she improved clinically, there was an increase in the evoked motor unit potential, the serum levels of some of the muscle enzymes decreased to normal, and the serum and tissue vitamin E levels increased significantly. It was concluded that treatment with high doses of vitamin E was responsible for the arrest of the usually progressive neuropathy and myopathy. PMID- 2981137 TI - Minor salivary gland tumors. A histologic and immunohistochemical study. AB - Clinical, histologic, and immunohistochemical data were reviewed and evaluated on 238 oral minor salivary gland tumors. Benign neoplasms accounted for 65% of the total. Pleomorphic adenomas were the most common of all neoplasms, and mucoepidermoid carcinomas were the most frequently encountered malignancies. Pleomorphic adenomas and adenoid cystic carcinomas stained positive for S-100 protein. Immunohistochemistry was believed to be of potential assistance in diagnosis of salivary gland tumors and in prediction of histogenesis. PMID- 2981138 TI - Sclerosing hemangioma of the lung. Immunohistochemical characterization of its origin as related to surfactant apoprotein. AB - Lung tissues from 13 patients with pulmonary sclerosing hemangioma were studied with antibody against surfactant apoprotein, Factor VIII-related antigen, or lysozyme. Surfactant apoprotein was detected in the cytoplasm of the cells lining cystic spaces and papillary projections. Surfactant apoprotein was found in a small number of stromal cells with abundant eosinophilic or clear cytoplasm and round to oval nuclei, which were characteristic in pulmonary sclerosing hemangioma as the main component. Surfactant apoprotein was also found in the stromal cells with small, dark nuclei similar to the lining cells. The lining and stromal cells contained neither Factor VIII-related antigen nor lysozyme. Our demonstration of surfactant apoprotein in these cells provides further support for the idea that pulmonary sclerosing hemangioma primarily consists of epithelial cells with differentiation to type II pneumocytes, as was deduced from ultrastructural investigations. PMID- 2981139 TI - Human papillomavirus infection of the esophagus. A clinicopathologic study with demonstration of papillomavirus antigen by the immunoperoxidase technique. AB - Papillomaviruses are known to be oncogenic in animals. In humans they are associated with benign squamous tumors (verruca, condylomata acuminata, and papillomas) in a variety of body sites. Human papillomavirus (HPV) infection of the esophagus, however, has not previously been documented. Recent reports of condylomatous changes in esophageal epithelium adjacent to esophageal carcinoma and the sporadic descriptions of esophageal papillomas in the literature for many years, lend credence to the assumption that HPV may affect the squamous mucous membrane of the esophagus. In the current study 75 cases, including 2 papillomas and 73 focal epithelial hyperplasia of the esophagus, were examined for histologic evidence of HPV infection as characterized by the presence of koilocytosis, giant and multinucleated cells, dyskeratosis, hyperkeratosis, acanthosis, papillomatosis, and anisonucleosis. Thirteen of the cases--the 2 papillomas and 11 of the focal epithelial hyperplasias--contained distinctive histologic evidence of HPV infection. The presence of HPV antigens was demonstrated by immunoperoxidase (IMPO) in the 4 of the 13 cases (31%). In the remaining cases the IMPO was equivocal in two and negative in seven. PMID- 2981140 TI - Familial adult T-cell leukemia. AB - Two siblings who developed adult T-cell leukemia (ATL) are presented. The patient and 7 of 26 healthy family members examined had the serum antibodies against ATL associated antigens (ATLA). This family study shows that two main routes of transmission of human T-cell leukemia virus (HTLV) may be involved: one is the route from parents to children and the other is horizontal transmission among spouses, especially from husband to wife; the anti-ATLA-positive family is considered to be a high-risk group for ATL. PMID- 2981141 TI - Radiation-induced lung fibrosis after treatment of small cell carcinoma of the lung with very high-dose cyclophosphamide. AB - Twenty-five previously untreated patients with small cell carcinoma of the lung were treated with cyclophosphamide 160 to 200 mg/kg (with autologous bone marrow support) followed by radiotherapy (4000 cGy) to the primary site and mediastinum. No other treatment was given until relapse occurred. Nineteen patients were assessable at least 4 months after radiotherapy; of these, 15 (79%) developed radiologic evidence of fibrosis, which was symptomatic in 14 (74%). The time of onset of fibrosis was related to the volume of lung irradiated. A retrospective analysis was made of 20 consecutive patients treated with multiple-drug chemotherapy and an identical radiotherapy regimen as part of a randomized trial. Radiologic and symptomatic fibrosis was one half as frequent (35%) as in the high dose cyclophosphamide group. Very high-dose cyclophosphamide appears to sensitize the lung to radiotherapy and promotes the production of fibrosis. PMID- 2981142 TI - Chemical carcinogens and inhibitors of carcinogenesis in the human diet. AB - The induction of cancer by chemicals as presently understood involves a series of steps, some of which require the passage of time. Many substances that are potent carcinogens in experimental animals are known to exist in nature and occur as part of the human diet. In addition, many of the substances that are known to inhibit experimental carcinogenesis also exist in the human diet. Thus, in addition to industrially produced carcinogens, humans can be presumed to have evolved in an environment that contains both carcinogens and anti-carcinogens. There is also a great deal of experimental and human epidemiologic data on the influence of lipids, proteins and carbohydrates on cancer incidence rates; however, much of those data are confusing and conflicting. PMID- 2981143 TI - Changes in distribution of hepatic blood flow induced by intra-arterial infusion of angiotensin II in human hepatic cancer. AB - Changes in the distribution of the hepatic blood flow induced by intra-arterial infusion of angiotensin II (AT-II) were studied in human hepatic cancers using extremely short-lived radioisotope (RI) (krypton 81 m [81mKr]; half-life, 13 seconds). After the start of continuous infusion of AT-II, the radioactivity of the tumor showed about a two-fold increase, whereas that of the nontumor region decreased to about one half as much as the level before the infusion. Consequently, the mean ratio of the arterial blood flow in the tumor region to that in the nontumor region (T/N ratio) increased to 3.30 (P less than 0.001). The T/N ratio showed a peak before the peripheral blood pressure reached the maximum, and thereafter tended to decrease. Intra-arterial infusion of AT-II raised the T/N ratio more obviously than did intravenous infusion of the drug, with less rise in the peripheral blood pressure. It is believed that intra arterial infusion chemotherapy with local use of AT-II enables better accessibility of chemotherapeutic drugs to tumors. PMID- 2981144 TI - A malignant cystosarcoma phyllodes with positive estrogen receptor and its heterotransplantation into nude mice. AB - We detected cytosol estrogen receptors in a malignant cystosarcoma phyllodes and succeeded in serial transplantations into athymic nude mice. The tumors of this transplantable strain (MC-3-JCK) have the same histologic features as the original tumors, and retain significant amounts of cytosol estrogen receptors. This strain appears to provide a useful experimental model for the study of biologic and therapeutic aspects of human cystosarcoma phyllodes of the breast. PMID- 2981145 TI - Influence of nutritional status on circulatory ribonuclease C levels in patients with cancer. AB - The influence of a variety of clinical and biochemical parameters on the activities in serum of ribonuclease (RNAse) selective for polycytidylic acid (RNAse C) were examined in 90 adult patients with cancer. The clinical data base determined on each patient included: RNAse C level, carcinoembryonic antigen (CEA) level, age, sex, race, presence (or absence of metastases, type of cancer, site of metastasis, renal function blood urea nitrogen [BUN], creatinine), hepatic function (bilirubin, alkaline phosphatase), and nutritional status (percent ideal body weight, percent weight loss, and albumin). Common tumor types studied included: colon (21), lung (18), breast (15), and hepatocellular carcinoma (10). For comparison, 175 nonmalignant control patients were studied to establish the normal range for RNAse. In patients with cancer, RNAse levels were increased in 57% and CEA levels were above 10 ng/dl in 36%. Although patients with BUN greater than 25 mg/dl or creatinine greater than 1.5 mg/dl were not entered on the study, nonetheless, RNAse was significantly (P less than 0.05) associated with both BUN and creatinine. Nutritional status also had an important influence on RNAse levels as both percent weight loss and percent ideal body weight were significantly (P less than 0.05) associated with circulatory RNAse: weight loss resulted in higher RNAse levels. These results account in part for the increased RNAse levels seen in those malignant conditions such as pancreatic and lung cancer commonly associated with weight loss in advanced stage. The possibility that circulatory RNAse C determination will provide a sensitive means for assessing nutritional status in cancer patients will require prospective evaluation. PMID- 2981146 TI - Granular cell tumor of the proximal esophagus. A rare disease. AB - In 1926, Abrikossoff described a tumor, usually benign, which only rarely appears in the esophagus. In a 40-year-old woman, the authors found a multicentric granular cell tumor which was localized in the cervical esophagus and in the subcutaneous tissues over the right scapula; it was removed surgically. Granular cell tumors causing stenosis of the upper esophagus have been described only four times in the literature. Characteristics and treatment are discussed. PMID- 2981147 TI - Value of open-lung biopsy in 87 immunocompromised patients with pulmonary infiltrates. AB - The authors performed a retrospective analysis of 87 consecutive immunocompromised patients who underwent open-lung biopsy at the University of Utah Medical Center, Salt Lake City, Utah, from January 1971 to June 1982. A specific histologic diagnosis was obtained in 62 (71%) of the patients, 33 of whom had infections. Pneumocystis carinii was the most common microbial pathogen (16 patients), but no cases have been observed since 1980 when the routine use of prophylactic trimethoprim/sulfa began. The other specific diagnoses included malignancy or drug-induced lung disease. Specific therapy was available for 52 patients, and in 33 cases, a change in therapy was necessary to treat according to the lung biopsy diagnosis. Forty-one patients received an adequate course of therapy and 27 (66%) of these improved clinically, including 16 of 26 patients with infections, 11 of 14 with malignancies, and 1 of 2 with a vasculitis. Among the subgroup of 33 patients for whom a new, specific therapeutic option was available as a result of the biopsy diagnosis 21 (64%) responded to the treatment. Eleven significant operative complications were encountered, but no deaths were attributable to the biopsy. An open-lung biopsy in immunocompromised patients is a relatively safe, accurate diagnostic procedure which frequently facilitates appropriate therapy and clinical improvement. PMID- 2981148 TI - Small bowel cancers, B-cell lymphatic leukemia, and six primary cancers with metastases and prolonged survival in the cancer family syndrome of Lynch. AB - The Cancer Family Syndrome of Lynch (CFS) is characterized by the frequent occurrence of multiple types of cancers, often adenocarcinomas of the colon, endometrium, and breast, at early ages in a pedigree pattern suggesting an autosomal dominant mechanism of inheritance. An unusual CFS pedigree is presented in which members exhibited rare cancers, six primary cancers, and prolonged survival. PMID- 2981149 TI - Persistent Epstein-Barr virus infection and a histiocytic sarcoma in a renal transplant recipient. AB - Lymphomas occurring in renal transplant recipients are mostly large cell non Hodgkin's lymphomas (B-cell-derived). A sarcoma with all morphologic, immunohistochemical, and ultrastructural characteristics of a tumor of the mononuclear phagocytic system (MPS) developed in a 23-year-old woman 1 year after renal transplantation. Anti-Epstein-Barr-virus antibody titers proved to be exceptionally high, even in pretransplant sera. Tumor-derived cells proved to be positive for Epstein-Barr nuclear antigen (EBNA), and hybridization showed multiple copies of Epstein-Barr virus (EBV)-DNA, suggesting a relationship between this tumor and EBV. More widespread use of immunochemical and histochemical diagnostic techniques might detect more cases, which, until now, have probably been diagnosed as B-cell-derived immunoblastic lymphomas. PMID- 2981150 TI - Tn, a carcinoma-associated antigen, reacts with anti-Tn of normal human sera. AB - Tn antigen is the immediate precursor of the carcinoma (CA)-associated T antigen; both are masked in non-CA tissues. Tn antigen was detected by absorption of human anti-Tn antibody in 46 of 50 primary breast CAs and in all 6 metastases originating from Tn-positive primary CAs. Thirteen of 25 (52%) anaplastic CAs, but only 2 of 15 (13%) well differentiated CAs had more Tn than T; 1 anaplastic CA had neither antigen. Eighteen of 20 benign breast lesions had no Tn; the 2 positive lesions were premalignant. All 19 breast CAs, studied immunohistochemically, reacted strongly with human polyclonal anti-Tn; benign or normal glandular tissues had minimal or no reactivity. Among live cancer cell lines, the most malignant sublines had more Tn than T on their cell surfaces. Preliminary studies with rodent monoclonal anti-Tn and anti-T antibodies gave immunohistochemical reactivity patterns similar to those of the polyclonal antibodies, but the former were less sensitive in absorption tests. Tn is a CA marker that promises to be useful in tumor detection. PMID- 2981151 TI - Lymphomas of the mucosa-associated lymphoid tissue. Signet ring cell lymphomas presenting in mucosal lymphoid organs. AB - Four cases of lymphoma are reported which were postulate to have developed from neoplastic transformation of the lymphoid cells of the mucosa-associated lymphoid tissue (MALT). This contention is primarily based on two observations: the peculiar "homing" tendency of these lymphomas and the immunoglobulin isotype. Both properties are characteristic of the lymphoid cells of the MALT. These lymphomas can also be identified by their histologic structure (signet ring cell lymphomas) and their presentation in an MALT organ. These lymphomas are probably separable from other lymphomas proposed to arise from lymphocytes of the gut associated lymphoid system (GALT). PMID- 2981152 TI - Hypercalcemia, unusual bone lesions, and human T-cell leukemia-lymphoma virus in adult T-cell lymphoma. AB - Extensive generalized and subperiosteal bone resorption was demonstrated in a patient with adult T-cell lymphoma and marked hypercalcemia of unclear pathogenesis. Antibody to the human T-cell leukemia-lymphoma virus (HTLV) was present in the serum of the patient, consistent with the recently reported association of adult T-cell lymphoma, hypercalcemia, and HTLV. The unique feature of this case was the presence of bone radiographic and pathologic findings consistent with hyperparathyroidism, in the absence of elevated parathormone levels. These findings contrast with the few previously reported cases of adult T cell lymphoma with hypercalcemia, which showed lytic, sclerotic, or osteoporotic bone lesions. The authors suggest that the patient's malignant T-lymphocytes may have produced an osteoclast-activating-factor-like substance or a parathormone like substance, which caused the striking bone changes. The exact role of HTLV in the pathogenesis of such cases remains to be determined. PMID- 2981153 TI - Breast cancer in Nigeria. AB - Breast cancer as seen in 116 Nigerian women over a 5-year period, 1974 to 1979, at the University of Benin Teaching Hospital, Benin, Nigeria, is presented. The disease occurs one decade earlier in Nigerians and is mainly a disease of premenopausal and perimenopausal women. Reflecting their relative youthfulness, more than 10% of them are pregnant or lactating on presentation. The most common histologic finding is anaplasia with very unfavorable histological grade. Breast cancer does not seem to have a different biologic behaviour in Nigerians, but it carries a truly bleak prognosis because many of the patients present with incurable disease that is close to its end stages. PMID- 2981154 TI - The Philadelphia (Ph) chromosome in leukemia. I. A new mechanism due to interstitial deletion and insertion in chronic myelocytic leukemia. AB - The Philadelphia (Ph) chromosome in leukemia is invariably derived from chromosome #22. A break occurs in the long (q) arm of chromosome #22 and, in every case observed until now, all of the material from that breakpoint through the telomere of chromosome #22 has been reciprocally translocated to another chromosome, most often chromosome #9. With the t(9;22) translocation, the oncogene c-sis moves from chromosome #22 onto 9q and the oncogene c-abl moves reciprocally from chromosome #9 onto 22q. We report a new mechanism for the genesis of the Ph chromosome in chronic myelocytic leukemia (CML) involving interstitial deletion of chromosome #22 with insertion of the deleted material into another chromosome: 46,XX,dir ins(11;22)(q13;q11q13). The distal portion of chromosome #22, including the telomere, appeared to have been retained in the Ph chromosome. There was no visible involvement of chromosome #9. This insertional deletion is of potential importance in evaluating the roles of oncogenes such as c-abl and c-sis in the Ph rearrangement in the origin of leukemia. PMID- 2981155 TI - A cytogenetic study of Wilms' tumor. AB - Cytogenetic studies were attempted on a sample of 33 Wilms' tumors using short- and long-term tissue culture techniques; most were studied after 2-13 wk in vitro. Abnormal karyotypes were found in 11 tumors, 10 of which had been treated previously with radiotherapy and/or chemotherapy. Insufficient growth of 17 tumors prevented cytogenetic analysis, and in 5 only normal karyotypes were found. Of the 11 tumors with abnormalities, 9 had modes within the diploid range, 1 was hypertriploid, and 1 showed hypodiploid variation. Except for chromosome #5, all chromosomes were involved in structural and numeric changes. Three tumors showed variation in the short arm of chromosome #11, and abnormalities of chromosomes #1, #3, #7, #9, #10, and #14 occurred in more than one tumor. A predisposition to Wilms' tumor is associated with a constitutional deletion of 11p13. From the present study, it is apparent that a similar cytogenetic change may play a primary role in the development of Wilms' tumor in normal individuals. PMID- 2981156 TI - Elevated growth factor levels in transformed mouse embryo cells treated with N,N dimethylformamide. AB - The transformed mouse embryo fibroblast cell line AKR-MCA, produces several transforming growth factor (TGF) activities which can be identified in cell extracts and serum-free conditioned medium. Treatment of these transformed cells with 1% N,N-dimethylformamide resulted in a more normal phenotype and an increased level of TGFs in cell extracts and conditioned medium. In addition, an 11-fold increase in an epidermal growth factor receptor-competing activity was observed in cell extracts and conditioned medium compared to control untreated cells. Fractionation of the conditioned medium on Bio-Gel P-100 showed that the same size classes of TGFs were present in N,N-dimethylformamide-treated as well as untreated cells. The increased EGF-receptor-competing activity was eluted in two peaks at Mr 6500 and Mr 4000. The Mr 6500 peak did not coelute with a TGF peak in the N,N-dimethylformamide-treated cells. PMID- 2981157 TI - Aberrant cyclic adenosine 3':5'-monophosphate metabolism in cultures of tumorigenic rat urothelium. AB - The cyclic adenosine 3':5'-monophosphate (cyclic AMP) metabolism of stratified normal rat urothelium propagated in vitro on a floating collagen matrix was characterized and used as a basis for identifying potential biochemical lesions in tumorigenic cell lines. The four neoplastic urothelial cell types studied (AY 27, AY-32, AY-33, and AY-34) were derived from Fischer 344 rats fed the carcinogen N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide. Epinephrine or prostaglandin E1 caused a rise in the cyclic AMP content of normal cultures which was potentiated in the presence of the cyclic nucleotide phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine or by forskolin, a diterpene activator of adenylate cyclase in intact cells. The expected, normal profile of cyclic AMP accumulation in response to beta-adrenergic receptor agonists was epinephrine greater than norepinephrine greater than phenylephrine. By every measure, the tumorigenic AY-27 cells demonstrated an overall decrease of functional adenylate cyclase activity. This was evidenced most by the low accumulation of cyclic AMP observed in response to forskolin. While prostaglandin E1 elicited a heightened cyclic AMP level in these cells, their vanishingly low response to catecholamines also suggested a potential lack of functional beta-adrenergic receptors. Cyclic AMP phosphodiesterase activity was elevated in soluble enzyme preparations obtained from cultures of AY-27 cells. Observations of AY-32 cells were diametrically opposite to the findings with AY-27 cells. In AY-32 cells, prostaglandin E1 receptors appeared to be functionally absent. The beta adrenergic receptor agonist response profile was abnormal in AY-32 cells. Norepinephrine produced a greater accumulation of cyclic AMP than epinephrine, and phenylephrine stimulated a much greater than normal response. Forskolin stimulation indicated an average level of adenylate cyclase activity in AY-32 cultures. Soluble preparations from AY-32 cells demonstrated a normal amount of cyclic AMP phosphodiesterase activity. AY-33 cells were comparable to normal urothelial cells in all respects save one. These tumorigenic cells had elevated levels of cyclic AMP phosphodiesterase activity. AY-34 cells, like AY-32 cells, were deficient in their responsiveness to prostaglandin E1. However, unlike the other tumorigenic lines, AY-34 cells had an excess of adenylate cyclase as demonstrated by their extraordinary responsiveness to forskolin. In addition, the accumulation of cyclic AMP in AY-34 cells in response to stimulation by epinephrine and norepinephrine, but not phenylephrine, was unusually great.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981158 TI - Estrogen 2- and 4-hydroxylase activity, catechol estrogen formation, and implications for estrogen carcinogenesis in the hamster kidney. AB - Estrogen 2- and 4-hydroxylase (ESH), a microsomal enzyme which mediates the formation of catechol estrogens, has been studied in the kidneys of castrated male Syrian hamsters, a species uniquely susceptible to induction of renal carcinomas by both steroidal and stilbene estrogens. The apparent Km for estrone was 17.0 microM, and Vmax was 0.5 pmol per mg protein per min for ESH in renal microsomes derived from castrated hamsters. Different steroidal estrogen substrates exhibited decreasing catechol formation with hamster kidney microsomal preparations in the following order: estrone greater than d-equilenin greater than 17 beta-estradiol greater than equilin greater than ethynyl estradiol greater than estriol. Except for beta-dienestrol, the stilbene estrogens revealed levels of catechol formation that were similar to 17 beta-estradiol. These findings provide a rationale for the weak carcinogenic activity of ethynyl estradiol, estriol, and beta-dienestrol, since they were poor substrates for hamster renal ESH and for the relatively potent carcinogenic activity of the distal metabolite of diethylstilbestrol, indenestrol B/A, which exhibited substantial levels of o-hydroxylation when used as a substrate. Interestingly, ESH activity was significantly greater in the hamster kidney compared to corresponding rat tissue, and catechol estrogen formation was found to be 2.5- to 19-fold higher in the hamster kidney compared to the rat, using various steroidal and stilbene estrogen substrates. Moreover, the finding that a 3.5- to nearly 6 fold decrease, compared to untreated levels, in catechol formation in kidneys but not in livers of alpha-naphthoflavone-exposed hamsters, depending on the steroidal or stilbene estrogen substrate used, is consistent with the belief that the catechol estrogen pathway is pertinent to events leading to estrogen-induced renal tumorigenesis in the hamster. PMID- 2981159 TI - Altered expression of cell surface membrane antigens in a common acute lymphoblastic leukemia-associated antigen-expressing neuroblastoma cell line (SJ N-CG) with morphological differentiation. AB - The expression of common acute lymphoblastic leukemia antigen (CALLA) on a human neuroblastoma cell line, SJ-N-CG, was demonstrated by indirect membrane immunofluorescence, complement-dependent cytotoxicity, and quantitative absorption, using two monoclonal antibodies (J-5 and BA-3) directed against CALLA. Immunoprecipitation of solubilized 125I-labeled membrane proteins from SJ N-CG cells with J-5 antibody revealed a protein with a molecular weight of 100,000 as determined on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Morphological differentiation of SJ-N-CG cells could be induced in the presence of 2.0 mM dibutyryl adenosine 3'-5'-cyclic monophosphoric acid for 10 days of culture. Changes in cell surface membrane antigens associated with morphological differentiation were studied by indirect immunofluorescence and complement-dependent cytotoxicity using a panel of seven monoclonal antibodies. Increases in the antigens recognized by BA-2 (detecting leukemia-associated antigen), anti-Thy-1, and antibody 390 (Thy-1 antigen) were found in "differentiated cells," while those detected by BA-1 (B-cell-associated antigen) and J-5 (CALLA) were unchanged. In contrast, the antitransferrin receptor defined by B3/25 was inhibited, and expression of B7/21-defined la-like antigen was not induced. Kinetic studies on antigenic alterations showed that the expression of BA-2-defined antigen rose on Day 2 and remained at the same level until Day 10. The expression of CALLA was not changed from Days 2 to 10. The augmentation of Thy-1 antigen was noted on Day 4 and reached the maximum on Day 10. These results show that dibutyryl adenosine 3'-5'-cyclic monophosphoric acid is capable of inducing phenotypic changes in SJ-N-CG cells. The changes of expression of some antigens on exposure of cells to dibutyryl adenosine 3'-5'-cyclic monophosphoric acid may enable us to have a greater understanding of the differentiation of neuroblastoma to a more mature ganglioneuroblastoma phenotype. PMID- 2981160 TI - Epstein-Barr virus infections in hairy cell leukemia patients in the presence of complement-dependent neutralizing antibody. AB - Immune system status was characterized in patients with hairy cell leukemia (HCL) with respect to explaining their chronic or recurrent infections with Epstein Barr virus. Measures of cellular immune responsiveness for a group of 11 HCL patients were, in general, decreased when expressed as the proportion of tested patients with values less than 2 S.D. below mean values for a group of 17 healthy adults: T-cell enumeration, seven of 13; mitogen responsiveness of phytohemagglutinin, 10 of 11; concanavalin A, 10 of 11; pokeweed mitogen, 10 of 11; B-cell responsiveness by anti-immunoglobulin immunobead stimulation, two of six; responsiveness to streptolysin O antigen, four of seven; mixed-lymphocyte reaction, six of seven; natural killer cell activity, six of eight. Specific immunity to Epstein-Barr virus was measured by complement-independent, antibody mediated virus neutralization (mean index for HCL patients being 56% of control value) and complement-dependent virus neutralization (98% of control value). We concluded that, in spite of depressed levels of immune responses measured with general, cellular assays, functional levels of complement-dependent virus neutralizing antibody were present in these HCL patients. PMID- 2981161 TI - Distinct activities of interferon-gamma, lymphokine and cytokine differentiation inducing factors acting on the human monoblastic leukemia cell line U937. AB - The human monoblast leukemia line U937 is growth inhibited and induced to express various characteristics of mature monocytes by lymphokines (LK) and other cytokines. Previous experiments have shown that interferon-gamma (IFN-gamma) is responsible for some but not all of the differentiation-inducing effects on U937. To determine the variety and specificity of activity, the following factors were studied: phytohemagglutinin-induced LK that contained IFN-gamma (100 units/ml); purified IFN-gamma; human colony-stimulating factor 1 (CSF-1); and conditioned medium(a) (CM) from the human bladder carcinoma cell line 5637 and the hepatoma cell line SK-HEP. LK preparations contained no colony-stimulating activity, whereas CM from 5637 and SK-HEP both contained granulocyte-macrophage CSF (3000 to 4000 units/ml) but no IFN activity. IFN-gamma is the major immunoglobulin G Fc receptor-inducing species within lymphokine, since anti-interferon-gamma antibody inhibited most of this activity. Other sources of Fc receptor-inducing activity were CM from SK-HEP and 5637 cell lines. Human CSF-1 when tested up to 800 units/ml was inactive for Fc receptor induction. LK induced the chemotactic peptide receptor, but this induction was due to factors other than IFN-gamma as anti-IFN-gamma antibody did not inhibit the induction, and purified IFN-gamma at a dose equivalent to that found in LK (100 units/ml) had no activity in the assay. SK-HEP and 5637 CM had strong chemotactic peptide receptor-inducing activity, but human CSF-1 was inactive up to 800 units/ml. Peroxide production after stimulation with phorbol myristic acid could be induced by LK, LK with anti IFN-gamma antibody, 5637, and SK-HEP treatment. IFN-gamma (100 units/ml) and CSF 1 (800 units/ml) were ineffective. Peroxide production was induced by IFN-gamma at concentrations above 1000 units/ml. The inducibility of several enzymatic activities was determined as additional measures of maturation. N Acetylglucuronidase was induced, for example, by LK, IFN-gamma, 5637 CM, and phorbol myristic acid. Alkaline phosphatase was induced by LK, IFN-gamma, dexamethasone, and phorbol myristic acid. 1,25-Dihydroxycholecalciferol was also examined and could induce most of the maturational markers examined. The results demonstrate that non-IFN cytokines from several sources have profound differentiation-inducing effects on monoblast leukemia cells in a pattern different from that of IFN-gamma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981162 TI - Non-Hodgkin's lymphoma after successful therapy of small cell lung carcinoma. AB - A patient is presented who developed a diffuse mixed lymphoma after successful treatment of small cell undifferentiated carcinoma of the lung. The patient was treated with cytotoxic chemotherapy and irradiation for the lung cancer, and had no recurrence during 6 years with no treatment. He then developed stage IV lymphoma. The relationship between these two cancers and the occurrence of treatment-induced second malignancies in small cell carcinoma are discussed. PMID- 2981163 TI - Simple, rapid determination of serum guanase activity with the Hitachi 736 automated discrete analyzer. AB - In this new method for determining serum guanase activity by use of the Hitachi 736-40 automated analyzer, serum is incubated with a mixture of xanthine oxidase, superoxide dismutase, and catalase; a reagent containing KCN, guanine, nitrotetrazolium blue, and Triton X-100 is added; and the increase in absorbance at 570 and 660 nm is measured for 2.4 min. Only 20 microL of sample is required, and results are linearly related to the activity concentration of guanase up to 30 U/L. Within-run and day-to-day precision (CV) was respectively 2.6 to 4.2% and 3.5 to 5.5% over 0-30 U of guanase activity per liter. The normal reference interval, as calculated from data on 40 healthy persons, is 0.1 to 2.2 U/L. Results correlate well (r = 0.997) with those by a kinetic method (Clin Chem 27: 560, 1981). The guanase activity of 150 samples can be measured within 1 h by this method. PMID- 2981164 TI - Cross reactivity of a high-Mr alkaline phosphatase isoenzyme with a commercial antiserum to intestinal alkaline phosphatase. PMID- 2981165 TI - Activity of serum adenosine deaminase in sarcoidosis. PMID- 2981166 TI - Zinc content of cellular components of blood: methods for cell separation and analysis evaluated. AB - Platelets, mononucleated cells, polymorphonucleated cells, and erythrocytes were separated from whole blood by use of discontinuous gradients of colloidal polyvinylpyrrolidone-coated silica ("Percoll"). We measured the zinc content of these cells by flame atomic absorption spectrophotometry, using a modified technique for micro-samples that obviated matrix interferences. Thus, results obtained by conventional flame atomic absorption and by the micro-method were identical. Inter-comparisons of separation methods indicated that separation of platelets and mononucleated cells by a two-gradient system of "Ficoll-Hypaque" (a synthetic polymer of sucrose) or Percoll was relatively poor, whereas there was a good separation when a tertiary gradient system of Percoll was used. The apparent zinc content of mononucleated cells depended on the degree of separation from the platelets, with contamination by platelets resulting in artificially high values for mononucleated cells. PMID- 2981167 TI - Successful immune reconstitution in severe combined immunodeficiency despite Epstein-Barr virus and cytomegalovirus infections. AB - Cytomegalovirus (CMV) and Epstein-Barr virus (EBV), frequently found in the acquired immune deficiency syndrome (AIDS), have been suspected of contributing to the latter immunodeficiency. The ability of normal HLA-identical sibling bone marrow to reconstitute an 8-month-old infant with severe combined immunodeficiency infected with these two viral agents is of interest. After presentation with severe mucocutaneous candidiasis, cavitary pulmonary disease, nodular cutaneous lesions, and hepatic abscesses containing acid-fast organisms, immunologic studies revealed lymphopenia, 1-3% T cells, and no lymphocyte responses to mitogens. Prior to transplantation, the infant's blood B lymphocytes grew spontaneously in culture, suggesting they were infected with EBV. Indeed, an appropriate antibody response to EBV was detected at 2 months post transplantation. At 3 weeks postgrafting, neutropenia and cholestatic jaundice developed without other signs of graft versus host disease. Liver biopsy demonstrated CMV but no EBV by DNA hybridization. There was evidence of T- and B cell function by 2 weeks postgrafting, including vigorous in vivo and in vitro responses to candida. Although the blood lymphocyte T4:T8 ratio was inverted at 2 weeks, it reverted to normal by 6 weeks post-transplantation. All clinical disease resolved by 8 months and karotyping revealed all T and B lymphocytes to be XX. Thus, despite infections with both CMV and EBV, complete immunologic reconstitution was achieved in this, the most severe of all genetically determined immunodeficiency conditions, arguing against these viruses having a major role in the failure of bone marrow transplantation in AIDS. PMID- 2981168 TI - Defective monocyte function in patients with systemic lupus erythematosus. AB - Peripheral blood adherent cells from patients with systemic lupus erythematosus (SLE) were shown to have markedly reduced phagocytic activity as compared to normal adherent cells or those from non-SLE patients receiving corticosteroid therapy. Both resting and phagocytosing monocytes showed decreased hexose monophosphate shunt and glycolytic activity. Mononuclear cells from SLE patients showed grossly impaired proliferative activity after NaIO4 activation. Furthermore, addition of SLE adherent cells to normal adherent cell-depleted lymphocytes decreased [3H]thymidine incorporation of the latter cells following NaIO4 treatment. Addition of normal adherent cells to SLE lymphocytes corrected the previous defect, indicating that an adherent abnormality is responsible for the defect in SLE mononuclear cell proliferation to NaIO4 activation. PMID- 2981170 TI - ACTH therapy in status asthmaticus. PMID- 2981169 TI - Modulation of the sodium periodate (NaIO4) response in systemic lupus erythematosus (SLE): effect of interleukin-1 (IL-1), interleukin-2 (IL-2), phorbol myristate acetate (PMA), and indomethacin. AB - The proliferative activity of peripheral blood mononuclear cells (PBMN) from patients with systemic lupus erythematosus (SLE) activated by sodium periodate (NaIO4) is greatly diminished. The effect of the cytokines Interleukin-1 (IL-1) and Interleukin-2 (IL-2) on the NaIO4 reaction was investigated. Addition of IL-1 resulted in partial restoration of the reaction of SLE PBMN to NaIO4, and a similar effect was demonstrated in the presence of phorbol myristate acetate (PMA). Addition of IL-2 to NaIO4 activated SLE PBMN, however, caused a marked improvement in their proliferative activity. The presence of indomethacin resulted in only a slight increase in [3H]thymidine incorporation by the NaIO4 treated SLE cells. The results suggest that the defect in the response of SLE PBMN cells to NaIO4 is due to inadequate availability of IL-1 and IL-2. Excessive production of prostaglandin in SLE might also contribute to the defective response to NaIO4 but does not appear to play a major role. PMID- 2981171 TI - Serum neuron-specific enolase. A marker for disease extent and response to therapy for small-cell lung cancer. AB - Serum neuron-specific enolase (S-NSE) levels in 43 newly diagnosed untreated patients with small-cell lung cancer (SCLC) were compared with levels in 35 adult controls, 14 patients with non-small cell lung cancer (N-SCLC), and nine patients with noncancerous lung disease (N-CLD). The S-NSE level was raised (greater than or equal to 16 ng/ml) in 28 of 43 patients with SCLC, six of 16 patients with limited stage SCLC, and 22 of 27 of those with extensive stage SCLC. Extensive stage patients with SCLC had a significantly higher mean S-NSE level (50 ng/ml) than did limited stage patients with SCLC (16 ng/ml). Mean S-NSE levels in patients with N-SCLC and in patients with N-CLD were respectively 11 and 7 ng/ml. Serial measurements performed on 19 patients between the three-day-courses of chemotherapy showed an excellent correlation between S-NSE and clinical evolution. In addition, S-NSE was measured during the first three-day course of chemotherapy in 13 other patients; among them, seven had S-NSE levels greater than or equal to 100 ng/ml (mean: 490 ng/ml); these seven patients were responders; the remaining six had S-NSE levels less than 100 ng/ml (mean 28 ng/ml): among them, only two were responders. Such S-NSE measurements during the first cytostatic course seem to reflect the importance of tumor burden and may be valuable as early indicators of the response rate to chemotherapy. PMID- 2981172 TI - Isolation of a line of immortal chicken embryo fibroblasts after transfection with the nuclei of Rous sarcoma virus-transformed Chinese hamster cells. AB - Secondary cultures of chicken embryo fibroblasts were transfected with purified nuclei from lysed cells of a clonal line of temperature-sensitive Rous sarcoma virus (tsRSV)-transformed Chinese hamster fibroblasts. After propagation for 3 months an established cell line designated ChR32 was obtained in one chicken cell culture. The cells of this line have been propagated so far for 18 months, whereas normal chicken embryo fibroblasts died after 2 months. The established cells were heteroploid with a diploid modal number of macrochromosomes and two Z chromosomes. No Chinese hamster chromosomes could be identified. Southern blot analysis of DNA from the uncloned ChR32 cells and the clones provided evidence that these established cells were, in fact, clonal in origin and contained full length RSV proviruses and no defective proviruses. Furthermore, they contained, at the 3' end proviral-cellular junction, Bg/II, HpaI, KpnI, SacI, and XbaI fragments of the same size as the Chinese hamster donor cells, suggesting that the cellular sequence adjacent to the provirus is of Chinese hamster origin. The cells after establishment were able to grow continuously at 37 degrees or 41 degrees C and produce a large amount of ts sarcoma virus particles. A corollary finding was that these virus particles were non-leaky for the transforming function at the non-permissive temperature. PMID- 2981173 TI - Cleavage of a 135 kD cell surface glycoprotein correlates with loss of fibroblast adhesion to fibronectin. AB - We have previously described a group of three plasma membrane glycoproteins that are recognized by an adhesion-disrupting antiserum and that are involved in fibronectin-mediated BHK cell adhesion. A peculiar property of these molecules is their resistance to tryptic digestion. We have now extended this study in the attempt to identify the active component within this group of molecules. SR/BALB mouse fibroblasts, used in this work, expose at their surface only two trypsin resistant glycoproteins, gp1 (150 K) and gp2 (135 K), that are recognized by the adhesion-disrupting anti-BHK serum. Controlled proteolysis of the cell surface in the presence of a reducing agent results in the loss of cell adhesion to fibronectin-coated substratum. gp2 is selectively cleaved under these conditions. Moreover, cells treated with trypsin and reducing agent can no longer adsorb the adhesion-relevant antibodies from the anti-BHK serum. These data indicate that gp2 plays a critical role in the adhesion of SR/BALB fibroblasts to fibronectin coated substratum, and that disulfide bonds are important in the conformation and function of this molecule. PMID- 2981174 TI - Lack of correlation between loss of anchorage-independent growth and levels of transformation-specific p53 protein in retinoic acid-treated F9 embryonal carcinoma cells. AB - It has been shown that differentiated derivatives of retinoic acid (RA)-treated F9 embryonal carcinoma cells become non-malignant. In the present study it is asked whether this loss of malignancy is due to cellular differentiation. Because the ability of cells to grow in suspension correlates with in vivo tumorigenicity, we determined the time course of the loss of this property, after RA treatment, with relation to the differentiation to parietal endoderm and the acquisition of normalcy in several common transformation-specific properties of F9 cells. Our results show that pretreatment with RA for 24 h caused 80% inhibition of anchorage-independent growth in F9 cells, and this inhibition reached its highest level (98%) after pretreatment with RA for 48 h and longer. However, all other observed transformation-related properties, and the levels of plasminogen activator (marker for parietal endoderm) remained unaltered at this early post-treatment stage. These observations suggest that the loss of malignancy is a relatively early event in the biochemical pathways involved in the RA-induced differentiation of F9 cells. Furthermore, our data show that the presence of elevated levels of p53 alone may not be sufficient to maintain the anchorage-independent growth and the rapid proliferation of F9 cells. PMID- 2981175 TI - Adenylate cyclase activity in zona-free mouse oocytes. AB - Zona-free mouse oocytes, prepared by either chemical or enzymatic treatment, possess adenylate cyclase activity, since forskolin elevated cAMP to similar levels in either these oocytes or in oocytes with intact zonae. In addition, it was shown that oocyte isolation conditions can affect 'basal' cAMP levels. PMID- 2981176 TI - Stimulating the proliferation of quiescent 3T3 fibroblasts by peptide growth factors or by agents which elevate cellular cyclic AMP level has opposite effects on motility. AB - The effects of some chemically defined growth factors on the locomotion of quiescent Swiss 3T3 fibroblasts have been studied. A computer digitiser has been used to facilitate recording the paths followed by cells in time-lapse films; this method allows 500 cell-hours to be recorded in 1 h of real time. Individual cells in the same culture vary widely in speed. This variation is not associated with the positions of the cells in the cell cycle; a small deceleration which seems to occur in G2 cannot account for any significant part of the variation seen. Nor is it related to the time elapsing before the cell divides, although this is equally variable; the speed and age at division of particular cells appear to be entirely independent of one another. Nevertheless, good reproducibility is seen between the mean speeds of large numbers of cells from the same type of culture. The mean speed of quiescent cells is less than 2 microns/h. A mixture of epidermal growth factor (EGF) and vasopressin, in the presence of insulin, is known to be a potent promoter of proliferation in this system. We have found it to increase speed to 30 microns/h. Agents which stimulate the cellular level of cAMP are also known to be potent promoters of proliferation in the presence of insulin. We have found these agents to be inhibitors of locomotion; several cycles of cell division take place while the cells move at a speed no greater than that seen in the presence of cytochalasin B (CB) or colchicine. These findings therefore give further support to the idea that there may be two different classes of growth-promoting factors, with major differences in their mode of action. They show that some members of these two different classes have opposite effects on motility. PMID- 2981177 TI - Inhibition of the activity of restriction endonucleases by spermidine and spermine. AB - Physiological concentrations (0.5-2.0 mM) of spermidine and spermine were observed to inhibit the digestion in vitro of plasmid pJDB 207 by the restriction endonucleases BamHI (EC 3.1.23.6), EcoRI (EC 3.2.23.13), HindIII (EC 3.1.23.20), HpaI (EC 3.1.23.23) and PstI (EC 3.1.23.31). The polyamines protected all the tested restriction sequences of DNA, since the activity of all endonucleases used was strongly inhibited. These results show the need for caution when using polyamines as experimental tools for recombinant DNA chemistry. PMID- 2981178 TI - Biological activities of human recombinant interferon alpha/beta targeted by anti Epstein-Barr virus monoclonal antibodies. AB - The requirement of high doses of interferon (IFN) during therapy severely restrict its application. Thus a model using an Epstein-Barr virus (EBV) membrane antigen (MA) specific monoclonal antibody (MAb) was developed to assess the feasibility of coupling minimal amounts of IFN to a MAb and specifically delivering the IFN to the target cells. Coupled IFN was first shown to retain fully both its anti-viral and anti-proliferative properties when tested on human tumor cell lines QIMR-WIL (EBV-MA+) and the U-266 (EBV-MA-). A series of in vitro pulsing experiments demonstrated the specific targeting of both the anti-viral and anti-proliferative properties of IFN to the EBV-MA+ QIMR-WIL cells and not EBV-MA- cell lines. PMID- 2981179 TI - tRNA binding to programmed ribosomes increases the ribosomal affinity for tuberactinomycin O. AB - The binding of 14C-labelled tuberactinomycin O was analysed in equilibrium dialysis cells. The ionic conditions and the concentration of the labelled drug used in the binding assays allowed the binding of just one drug molecule per non programmed ribosome. Under these conditions, the occupation of the ribosomal P site by deacylated tRNAPhe in the presence of poly(U) increased the amount of [14C]tuberactinomycin O bound by a factor of two. Kanamycin, gentamicin and neomycin reduced the binding of tuberactinomycin O, whereas chloramphenicol, tetracycline, streptomycin and puromycin had no effect. A stimulation of the binding of tuberactinomycin O was found upon addition of erythromycin. PMID- 2981180 TI - SV40 chromatin structure is not essential for viral gene expression. AB - The biological activity and the fate of SV40 DNA (minichromosomes, DNA I, DNA II, DNA III) were tested in culture cells by immunofluorescence staining and blot analysis. Following microinjection of 2-4 circular SV40 molecules (minichromosomes, DNA I, DNA II) into the cytoplasm or the nuclei of monkey and rat cells, T- and V-antigen synthesis was demonstrable in nearly every recipient cell. Only linear DNA induced T-antigen synthesis with a very low efficiency after cytoplasmic injection. This low activity correlates with a rapid degradation of DNA III in the recipient cells. Further modifications observed immediately after injection are relaxation of superhelical molecules and formation of high-Mr DNA. Assembly of the injected DNA into SV40 chromatin-like structure, however, occurred only late after early viral gene expression. PMID- 2981181 TI - The antilipolytic effect of insulin does not require adenylate cyclase or phosphodiesterase action. AB - Insulin antagonized the lipolytic actions of epinephrine in rat epididymal adipocytes when the phosphodiesterase inhibitor, Ro 20-1724, was present. Adipocytes were depleted of functional cAMP by inhibiting adenylate cyclase with N6-phenylisopropyladenosine in the presence of adenosine deaminase such that Ro 20-1724 no longer stimulated lipolysis. The cAMP analogs 8-thioisopropyl-cAMP or 8-thiomethyl-cAMP, which are resistant to phosphodiesterase hydrolysis, were subsequently added to bypass adenylate cyclase and phosphodiesterase action. Under these conditions, insulin antagonized the lipolytic effects of these analogs, even in the presence of Ro 20-1724. PMID- 2981183 TI - Melphalan potently substitutes the N-terminal Tyr of D-Ala2-Leu5-enkephalin methyl ester. AB - In search of an affinity label of the opioid receptor, the nitrogen mustard melphalan, Mel, was built into the peptide chain of D-Ala2-Leu5-enkephalin (DALE) methyl ester in different positions. We report now that in contrast to the previous observations that an intact Tyr in position 1 is essential for opioid activity [(1980) Annu. Rev. Pharmacol. Toxicol. 20, 81-110], substitution of Tyr by Mel did not result in a loss of the binding affinity. Mel1, Leu5-enkephalin OMe competed for the binding sites of [3H]naloxone as potently as DALE did; IC50 values for both compounds were 50 nM. Mel substitution has led to one order potency decrease in binding to the delta-sites. 0.5-1 microM of the compound irreversibly inactivates 50% of the binding sites of [3H]naloxone, and 5-10 microM of that of [3H]DALE. These results shed new light on the structural requirements established for opioid peptides. In addition, the new derivative can be used as an affinity label of the opioid receptor. PMID- 2981182 TI - Human mitochondrial aldehyde dehydrogenase inhibition by diethyldithiocarbamic acid methanethiol mixed disulfide: a derivative of disulfiram. AB - A derivative and possible physiological metabolite of disulfiram, diethyldithiocarbamic acid methanethiol mixed disulfide, is shown here for the first time to inactivate the mitochondrial low-Km isozyme of human aldehyde dehydrogenase (EC 1.2.1.3). By comparing inactivating effects of diethyldithiocarbamic acid mixed disulfides with thiols of increasing chain length evidence is provided that steric hindrance is the reason for lack of inhibition of the mitochondrial enzyme by disulfiram in vitro. Since methanethiol is a normal metabolite [(1983) Annu. Rev. Biochem. 52, 187-222] the results also suggest a mechanism by which aldehyde dehydrogenase is inhibited by disulfiram and diethyldithiocarbamic acid in vivo. PMID- 2981184 TI - Acquisition process of differentiation competence in Dictyostelium discoideum. AB - It was previously shown [K. Okamoto, J. Gen. Microbiol. 127, 301 (1981)] that Dictyostelium discoideum cells dissociated from early aggregates, but not aggregation competent cells obtained in a suspension culture, undergo prespore differentiation, when transferred into a medium containing glucose, albumin, and cAMP. Therefore, the former, but not the latter, is considered to have been acquired "differentiation competence." In the present work, the requirements for cells to acquire the differentiation competence are investigated with D. discoideum NC4 strain. On solid substratum, the incubation above a threshold density is absolutely required for this process, while cell aggregation itself is not essential. In suspension cultures, the competence is acquired only under hypertonic conditions. Inhibition of protein synthesis or depletion of cAMP does not affect the acquisition process of the competence. The requirement of hypertonic treatment was also investigated with several other D. discoideum strains. PMID- 2981185 TI - Molecular cloning of gene sequences transcriptionally regulated by retinoic acid and dibutyryl cyclic AMP in cultured mouse teratocarcinoma cells. AB - F9 mouse teratocarcinoma stem cells differentiate into primitive endoderm cells in response to retinoic acid (RA). A cDNA library, synthesized from RA and dibutyryl cyclic AMP-treated F9 teratocarcinoma cells, has been screened for gene sequences regulated by RA. By hybridization-selection and in vitro translation, the pcJ6, pcJ31, and pcF117 homologous mRNAs encode polypeptides of 40,000; 35,000-37,000; and 24,000 Da respectively. The pcI56 and pcI5 homologous mRNAs encode laminin B and collagen IV (S-Y. Wang and L. J. Gudas, 1983, Proc. Natl. Acad. Sci. USA 80, 5880-5884). Prior to RA addition, these gene sequences correspond to low-abundance mRNAs (less than 0.05% of total cellular mRNAs). Within 24 hr after the addition of RA (5 X 10(-7) M) to F9 cells, the expression of these sequences increases dramatically, and at 72 hr after drug addition, a 5- to 30-fold induction of these genes can be attained. Addition of lower concentrations of RA results in a smaller increase in the expression of these genes. If the F9 cells are treated with both RA (5 X 10(-7) M) and dibutyryl cyclic AMP (but2cAMP), the levels of mRNA specific for these five inducible genes are further increased by approximately 30- to 110-fold over those in the stem cells; but2cAMP alone does not induce the expression of these genes. The expression of J6 and J31, but not I56 (laminin) and 15 (collagen IV), is also regulated by RA in the P19 teratocarcinoma stem cell line, which differentiates into a fibroblastic cell type in response to RA. In vitro transcription experiments demonstrate that laminin and collagen IV are transcriptionally regulated by RA; but2cAMP also enhances the rate of transcription of these genes in F9 cells. PMID- 2981186 TI - Metastatic Wilms' tumor causing acute hepatic-vein occlusion (Budd-Chiari syndrome). AB - We describe a case of Wilms' tumor in a 6-yr-old girl which extended from the right kidney and completely obstructed the inferior vena cava and hepatic veins to the level of the right atrium, producing an acute Budd-Chiari syndrome. After an initial course of chemotherapy, the primary tumor was excised and, using cardiopulmonary bypass, the tumor thrombus was removed from the inferior vena cava and hepatic veins. The Budd-Chiari syndrome resolved and after triple-drug chemotherapy and radiotherapy under the National Wilms' Tumor Study-2 protocol the child has remained disease-free for 5 yr. PMID- 2981187 TI - Pruritus ani: could it be malignant? AB - The management of extramammary perianal Paget's disease should include a careful search for an associated carcinoma in the pelvic viscera by means of proctoscopy and x-ray. Such local malignancies as Bowen's disease, which may be pigmented and multifocal in this region, and invasive squamous cell carcinoma may first manifest themselves as pruritus ani. PMID- 2981188 TI - Isolation of intestinal mononuclear cells: factors released which affect lymphocyte viability and function. AB - Whether toxic or immunomodulatory factors are released during the collagenase digestion phase of the isolation of mononuclear cells from human intestinal mucosa was investigated by assessing the effect of the collagenase supernatant on the viability and natural killer activity of normal peripheral blood mononuclear cells. Three hours' incubation in collagenase supernatant suppressed natural killer activity by 25 +/- 4% and decreased the viability of peripheral blood mononuclear cells by 11 +/- 2%. The ability of collagenase supernatants to kill 51Cr-labelled peripheral blood mononuclear cells over four hours was assessed in 16 collagenase supernatants, eight of which produced lysis of 20 +/- 4%. There was no ultrastructural evidence of early degenerative changes in the viable intestinal mononuclear cells fresh from the isolation process or in peripheral blood mononuclear cells incubated in collagenase supernatant. Because prostaglandins are known to inhibit natural killer activity, PGE was measured in 20 collagenase supernatants by radioimmunoassay and found to be high at 27.5 +/- 4.0 ng/ml. Addition of indomethacin to the collagenase medium, however, failed to abolish the inhibitory effect of collagenase supernatant on natural killer activity and did not increase the natural killer activity of isolated intestinal mononuclear cells. The release of cytotoxic and immunomodulatory factors during the isolation of intestinal mononuclear cells indicates the necessity for careful assessment of the potential effects of the isolation process on any function being examined and casts doubt upon the relationship between in vitro findings and in vivo functional capabilities. PMID- 2981189 TI - Prognostic factors in malignant mesodermal (mullerian) mixed tumors of the uterus. AB - Clinicopathologic studies of 16 homologous and 19 heterologous malignant mesodermal (Mullerian) mixed tumors of the corpora uteri showed that homologous tumors were more common in black women than in white women; in the former, tumors were diagnosed at a younger age and in more advanced stages than in the latter. Fifty-five percent of women with clinical stage I disease had a higher surgicopathologic stage. The most significant prognostic factors were pathologic extent of disease and vascular invasion in the myometrium. There was no difference in outcome between homologous and heterologous tumors. Initial surgery for staging is essential for the adequate evaluation and treatment of these patients. PMID- 2981190 TI - Gestational trophoblastic disease in women aged 50 or more. AB - Twenty cases of gestational trophoblastic disease in women aged 50 or more are reported. The lesions were 7 hydatidiform mole (35%), 8 invasive mole (40%), and 5 choriocarcinoma (25%). The most common presenting symptom was abnormal vaginal bleeding. Three choriocarcinoma patients were postmenopausal and all of them had choriocarcinoma. None of the patients with hydatidiform mole or invasive mole died of the disease, but 4 of 5 choriocarcinoma patients died of the disease. Because of the high rate (56.3%) of malignant sequelae after molar evacuation, a primary hysterectomy for the treatment of hydatidiform mole in this age group is recommended. It is important to maintain a high level of suspicion for the diagnosis of gestational trophoblastic disease in the elderly women. PMID- 2981191 TI - Colposcopy for postmenopausal women. AB - The number of postmenopausal women colposcopically diagnosed has markedly increased during the last 15 years. In some of them, a clear colposcopic impression, required for planning an appropriate therapy, was disturbed by severe senile cervicocolpitis, when examined by the conventional technique using acetic acid. With a 5-year experience of 79 paired colposcopies, it was concluded that estrogen administration is necessary as a routine precolposcopic procedure in such cases and oral administration of conjugated estrogens at 1.25 mg/day for 14 days is adequate for this purpose. PMID- 2981192 TI - Late recurrence of gestational trophoblastic disease. AB - Persistent normal human chorionic gonadotrophin (hCG) levels for a period of 1 year after treatment is considered to be a reliable criterion of complete and sustained remission in patients with gestational trophoblastic disease. This is because such patients rarely require further therapy. Two patients are presented who developed recurrent disease after being in remission for 18 and 21 months. One of the patients initially had metastatic disease in the lungs; the other had tumor in the lungs and brain. Both of the patients have remained clinically free of disease after retreatment; one patient for 31 months and the other patient for more than 4 years. The management of the patients is presented with an emphasis on the requirement for long-term surveillance. Possible mechanisms to account for the reactivation of disease after a prolonged latency period are considered. PMID- 2981193 TI - Development and characterization of a human ovarian cancer cell line which clones without agar in vitro. AB - A new human ovarian cancer cell line has been initiated which clones without agar in vitro. The cell line has been characterized by growth of a tumor resembling the primary tumor in a nude mouse, by human lactic dehydrogenase isozyme pattern, by human karyotype, and by lack of contamination by other cell lines. Initial studies have demonstrated the presence of epidermal growth factor receptors in this cell line. The utility of this line for clonogenic studies is demonstrated by dose-response studies with doxorubicin, cisplatinum, and 13-cis-retinoic acid. PMID- 2981194 TI - Cosmid cloning of Rickettsia prowazekii antigens in Escherichia coli K-12. AB - Rickettsia prowazekii DNA was partially digested with Sau3A or HindIII, ligated with the cosmid vector pHC79, packaged in vitro, and transduced into Escherichia coli HB101. Cosmid cloning of Sau3A-digested rickettsial DNA yielded 1,288 ampicillin-resistant colonies; 798 cosmid clones resulted with HindIII-digested rickettsial DNA. Chimeric cosmid DNA was extracted from the latter gene bank, digested to completion with HindIII, and compared by agarose gel electrophoresis with a HindIII digest of rickettsial genomic DNA. The two digestion profiles were quite similar in their overall banding patterns, indicating that the clone bank was significantly representative of the rickettsial genome. When both clone banks were screened for expression of rickettsial antigens by enzyme-linked immunosorbent assay with goat anti-R. prowazekii serum, ca. 20% of the clones reacted positively. Two clones were randomly selected for more detailed analysis. Each contained a large chimeric plasmid (40.2 and 38.1 kilobases) which apparently yielded smaller deletion derivatives (13.6 and 12.6 kilobases) when transformed into an E. coli minicell strain. Each recombinant plasmid directed the synthesis of new protein species not observed in control minicells. One of the clones produced a 51,000-dalton protein in minicells, which comigrated with a protein reactive with anti-R. prowazekii serum. This protein was not present in negative controls. When antibodies to this protein were incubated with a Western blot of rickettsial total protein, they bound to a 52,000-dalton polypeptide. Hence, the cloned rickettsial gene product in E. coli corresponds to a protein of similar size in R. prowazekii. This study demonstrates the feasibility of cosmid cloning of rickettsial antigens in E. coli. PMID- 2981195 TI - Murine natural resistance to Trypanosoma lewisi involves complement component C3 and radiation-resistant, silica dust-sensitive effector cells. AB - The phenomenon of natural (innate) resistance is exemplified by the solid resistance of mice to infection with the rat-specific Trypanosoma lewisi. We provide more information about the mechanism of resistance in this model system. Resistance was not diminished in aged mice or reduced by ionizing radiation or splenectomy. There was no difference in resistance of C5-deficient mice (B10.D2/oSn and AKR) compared with their normocomplementemic counterparts (B10.D2/nSn and C57BL/6). Treatment of mice with cobra venom factor resulted in greatly prolonged survival of T. lewisi in mice. Combined treatment of mice with cobra venom factor and silica dust resulted in not only prolonged survival but also considerable intravascular growth of the parasite and resultant death of the mice. T. lewisi cells from irradiated donor rats, or after surface coat removal with trypsin, were eliminated by inoculated mice more efficiently than were parasites acquired from normal rats. Incubation of trypanosomes obtained from irradiated rats in normal rat serum restored their resistance to rapid elimination. From the results of these studies and other recent investigations, we have concluded that murine resistance to T. lewisi involves activation and binding of C3b by uncoated trypanosomes and concomitant participation of radiation-resistant, silica dust-sensitive effector cells (probably macrophages and neutrophils), and rat plasma proteins, integrated into the trypanosome surface coat, stabilize the coat, make it less susceptible to removal when the trypanosome is present in the mouse bloodstream, and thus delay exposure of underlying molecules (receptors?) that activate murine C3. PMID- 2981196 TI - Pulmonary macrophage function during experimental cytomegalovirus interstitial pneumonia. AB - Since cytomegalovirus (CMV) infections may alter host defense against a variety of pathogens, phagocytosis, oxygen uptake, and H2O2 release by pulmonary macrophages obtained from guinea pigs with acute CMV interstitial pneumonia were evaluated. Experimental animals were inoculated subcutaneously on day zero with 10(7.5) 50% tissue culture infective doses of virulent guinea pig CMV. Control animals received an uninfected salivary gland suspension. The animals were sacrificed on day 7; the tissues were cocultivated for virus isolation, and the lungs were lavaged to obtain pulmonary macrophages. CMV was isolated from buffy coat cells (96%), bone marrow cells (71%), whole lungs (77%), pulmonary macrophages (60%), and pulmonary granulocytes (49%). There was no significant difference between groups at sacrifice in the total number of macrophages obtained by pulmonary lavage or in the phagocytic activity of the macrophages in vitro. However, in CMV-infected animals, the maximum rates of O2 consumption in response to the soluble stimulus, phorbol myristate acetate, and the particulate stimulus, Staphylococcus aureus, were 47 and 55%, respectively, of the rates in uninfected controls. Total macrophage O2 consumption in CMV-infected animals was 32 and 37%, respectively, of control values in response to the same stimuli. In CMV-infected animals, the maximum rates of H2O2 release were 22% of those in simultaneous controls for both stimuli, and total H2O2 release was 30 and 25%, respectively, of that in controls in response to these stimuli. Such alterations in macrophage oxidative function may contribute to superinfection during CMV pneumonia. PMID- 2981197 TI - Oxidative metabolism of neonatal and adult rabbit lung macrophages stimulated with opsonized group B streptococci. AB - We compared the oxidative metabolism of alveolar macrophages (AM) from adult and neonatal (1- and 7-day-old) rabbits before and after their in vitro exposure to type Ia group B streptococci (GBS) opsonized with immune rabbit serum. Nonstimulated AM from 1-day-old, 7-day-old, or adult rabbits consumed O2 at a rate of 17 to 20 nmol/10(6) AM per 10 min under basal conditions and released minimal amounts of superoxide (O2-) into the medium. Approximately 80% of this basal respiration was of mitochondrial origin, based on its inhibition by NaCN. Exposure to GBS opsonized with immune rabbit serum stimulated O2 consumption approximately half as effectively in the neonatal AM as in the adult AM. Little O2- was released into the medium unless the cells were pretreated with dihydrocytochalasin B. Under such conditions, 1-day-old, 7-day-old, and adult AM released 3.6, 5.3, and 13.9 nmol of O2-/10(6) AM per 10 min, respectively. The uptake of opsonized GBS by 1-day-old AM was not affected by 1 mM NaCN, whereas phagocytosis by adult AM was substantially reduced under these conditions. Overall, our findings suggest that neonatal AM have less-well-developed postphagocytic oxidative metabolic responses and release less superoxide after exposure to opsonized GBS than do adult AM. They also demonstrate that the energy for phagocytosis is derived principally from mitochondrial metabolism in adult AM but not in neonatal AM. We conclude that metabolic differences between neonatal and adult AM may contribute to neonatal pulmonary susceptibility to GBS infections and account, in part, for the ability of GBS to succeed as neonatal pulmonary pathogens. PMID- 2981198 TI - Cloning of the gene for the common pathogenic Neisseria H.8 antigen from Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae DNA that encodes the pathogenic Neisseria H.8 common antigen was cloned in the lambda phage sep6. The recombinant phage, designated s6H.8, was detected immunologically with a monoclonal antibody that binds to the H.8 antigen. The gonococcal and s6H.8 forms of the antigen yielded identical partial proteolysis epitope maps. Neisseria species that did not manifest the H.8 antigen showed little or no DNA homology with s6H.8. This clone should facilitate investigation into the clinical utility of this antigen and determination of its possible function in gonococcal pathogenesis. PMID- 2981199 TI - Construction of a conjugative plasmid with potential use in vaccines against heat labile enterotoxin. AB - A conjugative plasmid with potential usefulness for vaccine strains was constructed. In the first step, a 5.9-kilobase DNA segment containing the two loci for the A and B subunits of heat-labile enterotoxin with a mutation in the gene for the A subunit was joined to the cloning vehicle pGA22, generating the nonconjugative plasmid pPMC4 with genes for resistance to tetracycline and chloramphenicol. In the second step, a segment of pPMC4 containing the genes for the A and B subunits, the gene for chloramphenicol resistance, and the replication genes of pGA22 was ligated to the genes for conjugal transfer of the F plasmid, generating the 54.9-kb plasmid pPMC5. Eleven porcine Escherichia coli isolates were tested as recipients for pPMC4 and pPMC5. For pPMC4, transformation and mobilization with a conjugative R plasmid were used to effect plasmid transfer. Only 1 of the 11 strains acted as a recipient in transformation. Mobilization with the R plasmid occurred with two strains, but the plasmids were altered during transfer. In contrast, pPMC5 was transferred with high frequency and unaltered to 9 of the 11 E. coli strains. Transconjugants from these nine matings produced high titers of the B subunit and no active heat-labile enterotoxin. Plasmid pPMC5 was stable in three porcine E. coli strains tested; plasmid pPMC4 was somewhat less stable in these strains. The method we describe for the construction of conjugative chimeric plasmids offers an opportunity for introducing genes with potential for immunization into bacterial strains that are suitable for colonizing the appropriate host sites. PMID- 2981200 TI - Hypoglycemia associated with nonislet cell tumor in 13 dogs. AB - Hypoglycemia associated with nonislet cell tumor was found in 13 dogs. In each dog, clinical signs were related directly to adrenergic and neuroglucopenic effects of hypoglycemia and included collapsing episodes, tremors, restlessness, weakness, and grand mal seizures that were responsive to glucose administration. Eight of the dogs had hepatocellular carcinoma; surgical resection of the tumor achieved remission of clinical signs in 3 of these dogs. Other hepatic tumors associated with hypoglycemia included leiomyosarcoma and hemangiosarcoma involving solitary lobes of the liver. Nonhepatic tumors included splenic hemangiosarcoma, diffuse metastatic melanoma, and salivary gland adenocarcinoma. PMID- 2981201 TI - Activation of in vitro transcription and topology of closed DNA domains. AB - Activation of in vitro transcription of otherwise inert DNA sequences by purified yeast RNA polymerase II has been observed following the introduction in closed DNA domains of fragments of various origin. This enhancer-like effect on the in vitro transcriptional capacity is only detected in negatively supercoiled DNA domains and is characterized for each chimaeric plasmid by the superhelical density (- sigma) at which a sharp transition toward activation takes place. We have analyzed the topological state (as defined by localization and evaluation of the relative occurrence of secondary structures sensitive to S1 endonuclease) of the activated closed domains as a function of the conditions that determine the transcriptional enhancer effect, i.e. superhelical density, size, and nature of the components of the domains. We observe that variations in transcriptional capacity coincide with a defined pattern of secondary structures. These observations support a cause-effect relation between topology and regulation of transcription. PMID- 2981202 TI - The formation of paranemic and plectonemic joints between DNA molecules by the recA and single-stranded DNA-binding proteins of Escherichia coli. AB - During the initial pairing events in the transfer of a strand from a linear duplex to a homologous single-stranded circular DNA by the recA and single stranded DNA-binding proteins of Escherichia coli, two types of structure are formed that are distinguishable by their stability in the presence of protein denaturants. One type which is resistant to 5.2 M guanidinium chloride is most likely a D-loop that depends only on heteroduplex base pairing for its stability. These D-loops form rapidly when the ends of the linear duplex are homologous with the single-stranded DNA but do not form when the ends are heterologous. The second type appears to require protein, in addition to base pairing, for stability since it is rapidly dissociated by treatment with 5.2 M guanidinium chloride. These unstable structures form even when the ends of the duplex are not homologous with the circular single-stranded DNA. The stability and topological properties of the stable and unstable structures are consistent with those of plectonemic and paranemic joints, respectively (Bianchi, M., Das Gupta, C., and Radding, C. M. (1983) Cell 34, 931-939). The plectonemic joints can be generated in situ from paranemic joints by the addition of a restriction enzyme that cleaves in the region of homology, thus producing free homologous ends. Omission of single-stranded DNA-binding protein results in a large decrease in the rate of formation of both paranemic and plectonemic joints. PMID- 2981203 TI - Properties of an interphotoreceptor retinoid-binding protein from bovine retina. AB - Washes and extracts of frozen and fresh cattle retina contain a water-soluble high-molecular-weight, retinoid-binding protein that is distinct from three other retinoid-binding proteins previously isolated from this tissue. The protein can be purified to apparent homogeneity from retinal homogenates by a combination of gel filtration, lectin, and ion-exchange chromatography. Overestimation of the protein molecular weight was observed in several systems involving migration of the protein through a porous network. The approximate molecular weight obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 140,000, a value consistent with those reported by other laboratories. However, a more detailed analysis using the method of Ferguson revealed the protein to behave anomalously relative to several proteins used as sodium dodecyl sulfate-polyacrylamide gel electrophoresis standards. The apparent radius of the native protein, estimated from calibrated gel filtration, corresponded to a globular protein with a molecular weight of 240,000-280,000, suggesting that the protein was a dimer. However, when the molecular weight of native interphotoreceptor retinoid-binding protein (IRBP) was determined by a method with no shape dependence, sedimentation equilibrium, a value of 131,700 +/- 3,900 g/mol, was obtained. Sedimentation equilibrium in a dissociating solvent (6 M guanidine HCl) yielded a molecular weight of the smallest component of 120,100 +/- 2,300 g/mol. The similarity of values for the denatured and native molecular weight by sedimentation equilibrium demonstrates that the protein is a monomer. In further support of this, no evidence for a dimer was observed in cross-linking experiments with dimethyl suberimidate. The sedimentation coefficient (S0(20),w = 5.73 +/- 0.15 S) and molecular weight from sedimentation equilibrium were employed to calculate the frictional coefficient and Stokes radius of IRBP (f/f0 = 1.64, Rs = 55 A). The high value of f/f0 of the protein provides a reasonable explanation for the over estimation of the molecular weight of native IRBP on gel filtration. Approximately 2 mol of exogenous all-trans- or 11-cis-retinol were bound per mol of protein (131,000). Approximately 7% of the binding sites were saturated with endogenous ligand (11-cis-retinol, 88%; all-trans-retinol, 12%) following isolation from partially bleached cattle eyes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981204 TI - The structure of type IX collagen. AB - We present a detailed analysis both of tryptic peptides and amino-terminal sequences of the subunits of two collagenous fragments (HMW and LMW) previously isolated from pepsin extracts of chicken cartilage (Reese, C.A., and Mayne, R. (1981) Biochemistry 20, 5443-5448). This analysis and a comparison with the nucleotide sequence of the cDNApYN1738 (Ninomiya, Y., and Olsen, B.R. (1984) Proc. Natl. Acad. Sci. U. S. A. 81, 3014-3018) shows that HMW and LMW are pepsin resistant fragments of a unique collagen composed of molecules with three different polypeptide chains (alpha-chains). This collagen has been assigned the type number IX, and the alpha-chain encoded by pYN1738 has been given the designation alpha 1 (IX). Type IX collagen contains three triple-helical domains and at least two sets of interchain disulfide bridges. At the amino and carboxyl ends are noncollagenous domains which do not appear to be homologous to amino and carboxyl propeptides of interstitial collagens. PMID- 2981205 TI - Electron transfer across posterior pituitary neurosecretory vesicle membranes. AB - Secretory vesicles from the neurohypophysis have a transmembrane electron carrier very similar to that found in adrenal medullary chromaffin granules. Two different tests show that ascorbic acid contained in the vesicles will reduce an external electron acceptor. First, reduction of cytochrome c or ferricyanide in the medium by a neurosecretory vesicle suspension can be followed spectrophotometrically. Second, the membrane potential (inside positive) generated by electron transfer can be monitored using the membrane potential sensitive optical probe Oxonol VI. As in chromaffin granules, this electron transfer is probably mediated by cytochrome b561. It may function to regenerate internal ascorbic acid and to provide reducing equivalents needed by the intravesicular amidating enzyme. PMID- 2981207 TI - Deletion of the terminal sequences from cytochromes c. AB - A 12-residue helical segment was deleted from the amino terminus of human heart cytochrome c by cleavage with cyanogen bromide. The resultant hemepeptide (13 104)H is rather insoluble in aqueous solution at neutral pH. Analysis of the crystallographic model with the terminal segment removed as well as qualitative spectral analysis of a transiently soluble sample of (13-104)H suggests that the hemepeptide retains little of the conformation of the native protein. By contrast, hemepeptide (1-80)H obtained by deletion of the carboxyl terminus is quite soluble at neutral pH. Spectral and hydrodynamic measurements suggest that the heme is encapsulated in an apolar cluster stabilized in part by axial ligation of the heme iron. PMID- 2981206 TI - GTPase activity of the stimulatory GTP-binding regulatory protein of adenylate cyclase, Gs. Accumulation and turnover of enzyme-nucleotide intermediates. AB - The GTPase activity of the stimulatory guanine nucleotide-binding regulatory protein (Gs) of hormone-sensitive adenylate cyclase was investigated using purified rabbit hepatic Gs and either [alpha-32P]- or [gamma-32P] GTP as substrate. The binding of [35S]guanosine 5'-O-(thiotriphosphate) (GTP gamma S) was used to quantitate the total concentration of Gs. 1) GTPase activity was a saturable function of the concentration of GTP, with Km = 0.3 microM. MgCl2 monotonically increased the activity. The maximum observed turnover number was about 1.5 min-1. 2) During steady-state hydrolysis, 20-40% of total Gs could be trapped as a Gs-GDP complex and 1-2% could be trapped as Gs-GTP. The hydrolysis of Gs-GTP to Gs-GDP occurred with t 1/2 less than or equal to 5 s at 30 degrees C and t 1/2 approximately 1 min at 0 degrees C. Hydrolysis of Gs-GTP was inhibited by 1.0 mM EDTA in the absence of added Mg2+. 3) The rate of formation of Gs-GDP and the initial GTPase rate varied in parallel as functions of the concentrations of either GTP or MgCl2 (above 0.1 mM Mg2+). The ratio of the rate of accumulation of Gs-GDP to the GTPase rate was constant at 0.3-0.4. 4) The rate of dissociation of assayable Gs-GDP was biphasic. The initial phase accounted for 60-80% of total assayable Gs-GDP and was characterized by a t 1/2 of about 1 min. 5) Lubrol 12A9 potently inhibited the GTPase reaction and the dissociation of Gs-GDP in parallel, and inhibition of product release may account for the inhibition of steady-state hydrolysis. 6) The beta and gamma subunits of Gs markedly inhibited the dissociation of GDP from Gs in contrast to their ability to stimulate the dissociation of GTP gamma S. 7) GDP, GTP gamma S, and guanyl-5'-yl imidodiphosphate (Gpp(NH)p) competitively inhibited the accumulation of Gs-GDP. GTP gamma S and Gpp(NH)p inhibited the GTPase reaction noncompetitively, GDP displayed mixed inhibition, and Pi did not inhibit. These data are interpretable in terms of the coexistence of two specific mechanistic pathways for the overall GTPase reaction. PMID- 2981208 TI - Complete amino acid sequence of the ubiquinone binding protein (QP-C), a protein similar to the 14,000-dalton subunit of the yeast ubiquinol-cytochrome c reductase complex. AB - The amino acid sequence of the ubiquinone binding protein (QP-C) in the cytochrome bc1 region of the mitochondrial electron transfer chain was determined by analysis of peptides obtained by cyanogen bromide cleavage and staphylococcal protease digestion of succinylated derivatives. It was found to consist of 110 amino acid residues and its amino terminus to be blocked by an acetyl group, as determined by mass spectrometry of the amino-terminal peptide and a comparison with peptides chemically synthesized on high-performance liquid chromatography. The molecular weight of this ubiquinone binding protein including the acetyl group was calculated to be 13,389. The predicted secondary structure of QP-C has alpha-helical content of about 50% and QP-C was classified as an "all-alpha" or "alpha + beta" protein. This is the first report describing the amino acid sequence of the ubiquinone binding protein. A comparison of this sequence with that of the 14-kDa subunit of the yeast ubiquinol-cytochrome c reductase complex from the nucleotide sequence showed these two sequences to be quite similar. PMID- 2981209 TI - Powder and single-crystal electron paramagnetic resonance studies of yeast cytochrome c peroxidase and its peroxide and its peroxide compound, Compound ES. AB - Electron paramagnetic resonance absorption spectrum of ferric cytochrome c peroxidase exhibited a mixture of high- and low-spin compounds. The principal values and the eigenvectors of the g-tensor for the low-spin species were determined by single-crystal EPR spectroscopy at 77 K. The powder EPR spectra of the peroxide compound, Compound ES, were measured at S-, X-, and Q-band microwave frequencies. Careful examination at 77 K showed a narrow free radical-like signal at g = 2.004 with hyperfine structures accompanied by a broad signal spreading on both low- and high-field sides. Single-crystal EPR analyses of Compound ES clearly demonstrated that there exist at least two different radical species: one is isotropic with hyperfine structure at g = 2.004 and the other exhibits an axially symmetric signal at 5 K and broad signal centered at g = 2.004 at 77 K, respectively. The principal values and the eigenvectors of the g-tensor for the axially symmetric signal were determined: g(parallel) = 2.034 and g(perpendicular) = 2.006, 1.999. The orientation of the unique axis (g(parallel)) was found to be identical to that of the heme normal. A new radical signal with complicated hyperfine structures in the g = 2.004 region was observed upon illumination of Compound ES at both 5 and 77 K. The photoinduced species grew effectively by the illumination light around 500 nm. On warming to -80 degrees C, the photoinduced signal was reversibly brought back to the original radical species of Compound ES via an intermediate species. From these results, we have proposed the possible sites for the free radical centers in Compound ES. PMID- 2981211 TI - Restriction nuclease digestions driven to completion by Escherichia coli RNA polymerase and T4 gene 32 protein. AB - Restriction enzyme digestions of large scale DNA preparations often do not go to completion. This is due to product inhibition by the newly generated ends of the digested DNA. The addition of exogenous proteins that bind tightly to the free ends of DNA or to single-stranded DNA will relieve this inhibition. We show that a considerable savings on restriction nucleases can be attained by the addition of RNA polymerase or T4 gene 32 protein in stoichiometric amounts to the newly produced DNA ends. PMID- 2981210 TI - pH dependence of hormonal regulation of gluconeogenesis and urea synthesis from glutamine in suspensions of hepatocytes. AB - Hepatocytes from rats deprived of food for 48 h synthesized glucose and urea from glutamine at a rate which, at pH 7.3, was markedly stimulated (175-250%) by dibutyryl cAMP, phenylephrine, and norepinephrine, in agreement with previous investigators. These effectors also stimulated respiration, elevating ATP production by the amount required for the increase in glucose and urea synthesis. Both the basal and stimulated rates were strongly pH dependent with maxima in the region of pH 7.2-7.6 (urea synthesis) and 7.2-7.5 (glucose synthesis) and declined rapidly on either side of these pH values. The inhibitions at acid and alkaline pH were neither due to lack of energy nor to limitation in glutamine uptake. The intracellular concentrations of aspartate, glutamate, and glutamine were lower at pH 6.7 than at pH 7.3 and were differently affected by dibutyryl cAMP and phenylephrine at the two pH values investigated. When calcium was omitted from the suspending medium, the basal rates of glucose and urea production were decreased as was stimulation by the effectors, phenylephrine completely, and the others partially. The stimulations by phenylephrine and dibutyryl cAMP were additive under all conditions tested. The pattern of metabolite changes indicates that although both effectors stimulated glutaminase and increased supply of aspartate to the argininosuccinate synthetase, dibutyryl cAMP gave greater activation of glutaminase whereas the adrenergic agonists gave greater stimulation of later steps on the biosynthetic pathways. It may be physiologically important than at acid pH both ureagenesis and gluconeogenesis are severely suppressed and cannot be effectively stimulated by the major hormonal regulators of these pathways. PMID- 2981212 TI - Intron positions are conserved in the 5' end region of myosin heavy-chain genes. AB - We have determined the 5' end sequence of the rat embryonic skeletal muscle myosin heavy-chain (MHC) gene comprising the first three amino-terminal coding exons. Comparison with the corresponding regions of the rat ventricular alpha and the nematode Caenorhabditis elegans unc-54 MHC genes (Mahdavi, V., Chambers, A.P., and Nadal-Ginard, B. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 2626-2630; Karn, J., Brenner, S., and Barnett, L. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 4253-4257) shows that the degree of amino acid sequence conservation increases from the first to the third exon. Intron positions between these exons are maintained in all three genes studied, whereas size and sequence of corresponding introns are highly divergent. In contrast to the rat MHC genes where the coding region is highly split throughout its entire length, only the 5' end region is frequently interrupted by introns in the nematode gene indicating the potential importance of these introns in gene structure and expression. The occurrence of "preferential" intron positions in the MHC genes suggests the existence of a highly split ancestral MHC gene from which different evolutionary lineages removed and/or added specific sets of introns. PMID- 2981213 TI - Characterization of a membrane-associated receptor from rat sinusoidal liver cells that binds formaldehyde-treated serum albumin. AB - When treated with formaldehyde, serum albumin is known to be taken up and degraded by sinusoidal liver cells via adsorptive endocytosis. The present study aimed at characterization and identification of the membrane-associated receptor on rat sinusoidal liver cells. Kinetic studies of binding of 125I-labeled formaldehyde-treated serum albumin (125I-f-Alb) with the membranes of sinusoidal liver cells demonstrated the presence of specific, high-affinity, saturable membrane-bound receptors with an apparent Kd = 8 micrograms of f-Alb/ml and the optimal pH at around 8.0. The 125I-f-Alb binding to the membranes was not inhibited by either native albumin, asialofetuin, methylamine-treated alpha 2 macroglobulin, mannan, or immune complexes. The binding process exhibited independence of calcium and susceptibility both to heat treatment and to destruction by proteases. The binding was inhibited by concanavalin A and the inhibition was effectively reversed by the presence of alpha-methyl-D-glucoside, a haptenic inhibitor for this lectin, indicating the glycoprotein nature of the receptor. The binding protein was extracted from the membrane preparations with octyl beta-D-glucopyranoside and immunoprecipitated by anti-ligand antibody as a complex with the ligand. Sodium dodecyl sulfate-gel electrophoresis of the immunoprecipitate revealed two polypeptide chains with molecular weights of approximately 53,000 and 30,000, respectively. PMID- 2981214 TI - Purification of a receptor for formaldehyde-treated serum albumin from rat liver. AB - A membrane-associated receptor involved in a specific uptake of formaldehyde treated serum albumin (f-Alb) was purified from rat livers by Triton X-100 solubilization of a 105,000 X g membrane preparation and affinity chromatography on an f-Alb-Sepharose column. The purified receptor exhibited Mr = 125,000, consisting of two noncovalently linked glycoprotein components with Mr = 53,000 and Mr = 30,000, respectively. Incubation of 125I-receptor with f-Alb, but not with native albumin, resulted in a marked shift of pI value from 5.9 to 5.1, reflecting the presence of a specific ligand-receptor interaction. The receptor incorporated into liposomes displayed a saturable binding to 125I-f-Alb and the binding was effectively replaced by the presence of unlabeled f-Alb, with binding parameters being similar to those obtained from 125I-f-Alb binding to the sinusoidal liver cell membrane (Horiuchi, S., Takata, K., and Morino, Y. (1985) J. Biol. Chem. 260, 475-481). Reaction of anti-f-Alb receptor antibody with extracts of sinusoidal cells resulted in a specific precipitation of two proteins whose molecular weights were identical to those for the purified receptor. The anti-receptor IgG fraction effectively blocked 125I-f-Alb binding to the sinusoidal cell membranes. These results indicate that the purified protein represents the membrane-associated receptor which is presumably involved in a specific uptake of this ligand from the circulation. PMID- 2981215 TI - Molecular properties of neurotensin receptors in rat brain. Identification of subunits by covalent labeling. AB - Neurotensin binding sites in rat brain synaptic membranes were specifically and covalently labeled by two methods. In the first, a photoreactive and highly radioactive analogue of neurotensin, 125I-labeled azidobenzoyl[Trp11]neurotensin, was synthesized and used to photoaffinity label neurotensin receptors. In the second, the reversible association between neurotensin receptors and 125I labeled[Trp11]neurotensin, a radioactive but nonphotoreactive analogue of neurotensin, was made irreversible by means of disuccinimidyl suberate, a bifunctional cross-linking reagent. Analysis of synaptic membranes by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that using both methods the same two protein bands with apparent molecular weights of 49,000 and 51,000 were specifically labeled. Identical results were obtained with or without reduction of the photolabeled membranes by beta mercaptoethanol before electrophoresis. Variation of the ligand concentration did not modify the relative labeling intensities of the two bands, indicating that the high- and low-affinity neurotensin binding sites previously detected in rat brain synaptic membranes have similar molecular structures. These results indicate that neurotensin receptors in rat brain may be composed of two different protein subunits with similar molecular weight of about 50,000, that are linked together by noncovalent bonds. PMID- 2981216 TI - Scavenger function of sinusoidal liver cells. Acetylated low-density lipoprotein is endocytosed via a route distinct from formaldehyde-treated serum albumin. AB - Chemically modified proteins such as acetylated low-density lipoprotein (acetyl LDL) and formaldehyde-treated serum albumin (f-Alb) infused intravenously are known to undergo receptor-mediated endocytosis by sinusoidal liver cells, major intravascular scavenger cells in vivo. The aim of the present study was to elucidate whether the endocytic uptake of acetyl-LDL and f-Alb is mediated by the same receptor or not. Experiments on the binding of 125I-acetyl-LDL to isolated rat liver sinusoidal cells revealed the presence of a specific, high-affinity, saturable, membrane-associated receptor with an apparent Kd = 7 micrograms of the ligand at 0 degrees C. Unlabeled acetyl-LDL effectively inhibited 125I-f-Alb binding to the cells. By contrast, the binding of 125I-acetyl-LDL to the cells was affected neither by unlabeled f-Alb nor by the antibody raised against the f Alb receptor. These results indicate that the scavenger receptors for these two ligands are distinct from each other but similarly sensitive to polyanionic compounds. PMID- 2981217 TI - Coordinate regulation of the levels of type III and type I collagen mRNA in most but not all mouse fibroblasts. AB - A mouse genomic clone was isolated by cross-hybridization with a DNA fragment which codes for the NH2-propeptide of chick alpha1(III) collagen. The region of cross-hybridization within the mouse clone was localized, its sequence determined, and an exon coding for the NH2-propeptide of mouse alpha1(III) collagen was identified. This DNA fragment hybridizes to an RNA species of approximately 5300 nucleotides, slightly larger than the major alpha2(I) collagen RNA species. The mouse type III collagen probe was used to examine the effect of transformation on alpha1(III) collagen RNA levels in mouse fibroblasts. The levels of type III and type I collagen mRNA levels were compared in control and sarcoma virus-transformed murine cell lines, as well as in NIH 3T3 cells transformed by members of the human ras oncogenes. The levels of type III RNA decreased about 10-15-fold in Moloney sarcoma virus-transformed cells and in a cell line transformed with a v-mos-containing plasmid, but showed only a 50% decrease in a Kirsten murine sarcoma virus-transformed BALB 3T3 cell line, and increased 4-fold in a Rous sarcoma virus (RSV)-transformed BALB 3T3 cell line. In contrast, the levels of alpha2(I) collagen mRNA are 8- to 10-fold lower in all these cell lines when compared to untransformed cells. NIH 3T3 cells transformed with two human ras oncogenes showed decreased levels of alpha2(I) and alpha1(III) mRNAs. In contrast to the RSV-transformed mouse cell line, RSV-transformed chick embryo fibroblasts contained much smaller amounts of type III RNA than control chick embryo fibroblasts. We conclude that the levels of alpha1(III) and alpha2(I) collagen mRNA are often but not necessarily coordinately regulated by transformation in mouse cells. PMID- 2981218 TI - Purification and characterization of enzymes from Euglena gracilis that methylate methionine and arginine residues of cytochrome c. AB - Two forms of cytochrome c-specific methyltransferases from Euglena gracilis were purified approximately 100- and 50-fold, respectively, using DEAE-cellulose and gel-filtration chromatography. The methylation product of enzyme I was identified as S-methylmethionine and that of enzyme II as NG-monomethylarginine. Both enzymes were located in the cytosol and exhibit maximum activity at pH 7.0. Among the various proteins tested as substrates, the enzymes were highly specific toward cytochrome c. Various types of histones, in particular, were not modified by either enzyme. The molecular weights of enzyme I and II were 28,000 and 36,000, respectively. Various S-adenosyl-L-homocysteine analogs were tested for their inhibitory activity toward the enzymes. Only the D- and L-isomers of S adenosylhomocysteine and sinefungin were significantly inhibitory. The Ki values for S-adenosyl-L-homocysteine were 8.13 X 10(-6) and 1.17 X 10(-5) M for enzyme I and II, respectively. Two-dimensional peptide mapping revealed the methylation site of enzyme I to be the methionine residue at position 65 while that of enzyme II to be the arginine residue at position 38. The methylation of either methionine or arginine residues by enzyme I and II, respectively, lowers the isoelectric point (pI) of cytochrome c: 9.23, 9.33, and 10.06 for S methylmethionine-, NG-monomethylarginine-modified, and unmodified cytochrome c, respectively. PMID- 2981219 TI - cGMP phosphodiesterase in rod and cone outer segments of the retina. AB - Immunochemical, chromatographic, and sodium dodecyl sulfate gel electrophoresis studies suggest that immunologically related but distinct cyclic GMP phosphodiesterases are present in rod and cone outer segments of the retina. Immunocytochemical studies demonstrated that one monoclonal antibody (ROS-1) recognized a determinant present in both rod and cone outer segments, while another monoclonal antibody (ROS-2) only recognized rod outer segments. At least two peaks of phosphodiesterase activity could be separated by high-performance anion-exchange chromatography of retinal extracts. Both peaks were recognized by ROS-1. None of the first peak and only 80% of the second broad peak of activity were recognized by ROS-2. High-performance liquid chromatography profiles from human fovea and several other types of cone-enriched retina showed that most of the activity was contained in the first peak, suggesting that this activity was derived from cone outer segments. Conversely, the phosphodiesterase in rod enriched preparations migrated predominately in the second peak. Sodium dodecyl sulfate-gel electrophoresis indicated that this first peak contained a single large immunoreactive polypeptide (alpha') that migrated with the same mobility as a phosphorylase b standard and was distinct from the more rapidly migrating large immunoreactive polypeptides (alpha and beta) present in a broad second peak. The second peak could be further separated into a first part that contained a doublet of two immunoreactive polypeptides (alpha and beta) that migrated faster than phosphorylase b and a later part that contained only the most rapidly migrating polypeptide (beta). All of the peaks could be activated by histone or transducin:GTP, implying that all contained a small 11-kDa inhibitory subunit (gamma) of the enzyme. Since the larger (alpha') and smaller (beta) immunoreactive polypeptides could be completely separated from the alpha polypeptide and from each other, yet still retain the ability to be activated by histone or transducin, the data suggest that only a single species of polypeptide inhibitor complex (e.g. alpha' gamma, alpha gamma, or beta gamma) was required for histone or transducin:GTP activation. PMID- 2981220 TI - Rho-dependent transcription termination at lambda R1 requires upstream sequences. AB - In order to define the DNA sequence requirements for rho-dependent transcription termination, we have constructed and analyzed two classes of mutants affecting the cro-tR1 region of phage lambda: one class contains progressively larger deletions in the region upstream of the rho-dependent terminator tR1, and the other consists of small insertions or deletions generated by the linker scanning procedure. We show that rho-dependent termination at lambda tR1 in vitro requires contribution from sequences far upstream of the sites of termination. No single linker scanning mutant completely abolishes transcription termination, indicating that there exists no unique sequence that serves as an obligatory rho utilization site. Our results suggest that rho can interact with a large portion of the RNA transcript, including sequences that would be translated in vivo. Moreover, rho action appears to require a minimal transcript length, about 150-190 nucleotides for the tR1 termination sites. We also examine the effect of the auxiliary transcription factor NusA on transcription of these mutants, and we provide support for the proposal that specific effects of NusA on transcription termination require a sequence known as "boxA" (Olson, E.R., Flamm, E.L., and Friedman, D.I. (1982) Cell 31, 61-70). PMID- 2981221 TI - Cloning and structure of the gene for the subunits of aspartokinase II from Bacillus subtilis. AB - A library of Bacillus subtilis DNA in lambda Charon 4A (Ferrari, E., Henner, D.J., and Hoch, J.A. (1981) J. Bacteriol. 146, 430-432) was screened by an immunological procedure for DNA sequences encoding aspartokinase II of B. subtilis, an enzyme composed of two nonidentical subunits arranged in an alpha 2 beta 2 structure (Moir, D., and Paulus, H. (1977a) J. Biol. Chem. 252, 4648 4654). A recombinant bacteriophage was identified that harbored an 18-kilobase B. subtilis DNA fragment containing the coding sequences for both aspartokinase subunits. The coding sequence for aspartokinase II was subcloned into bacterial plasmids. In response to transformation with the recombinant plasmids, Escherichia coli produced two polypeptides immunologically related to B. subtilis aspartokinase II with molecular weights (43,000 and 17,000) indistinguishable from those found in enzyme produced in B. subtilis. Peptide mapping by partial proteolysis confirmed the identity of the polypeptides produced by the transformed E. coli cells with the B. subtilis aspartokinase II subunits. The size of the cloned B. subtilis DNA fragment could be reduced to 2.9 kilobases by cleavage with PstI restriction endonuclease without affecting its ability to direct the synthesis of complete aspartokinase II subunits, irrespective of its orientation in the plasmid vector. Further subdivision by cleavage with BamHI restriction endonuclease resulted in the production of truncated aspartokinase subunits, each shortened by the same extent. This suggested that a single DNA sequence encoded both aspartokinase subunits and provided an explanation for the earlier observation that the smaller beta subunit of aspartokinase II was highly homologous or identical with the carboxyl-terminal portion of the alpha subunit (Moir, D., and Paulus, H. (1977b) J. Biol. Chem. 252, 4655-4661). A map of the gene for B. subtilis aspartokinase II is proposed in which the coding sequence for the smaller beta subunit overlaps in the same reading frame the promoter distal portion of the coding sequence for the alpha subunit. PMID- 2981223 TI - Localization of tyrosine kinase-coding region in v-abl oncogene by the expression of v-abl-encoded proteins in bacteria. AB - A series of plasmids containing different segments of the v-abl oncogene have been constructed to express different portions of the v-abl protein in bacteria. The tyrosine kinase activity of these proteins was determined by an in vitro assay employing histones or angiotensin II as substrates for the v-abl-encoded tyrosine kinase. These experiments show that the 5'-1.2 kilobases of v-abl is necessary and sufficient to produce an active tyrosine kinase which is functional as a monomeric soluble protein. The kinase-coding region corresponds to the minimal region of v-abl required for the transformation of fibroblasts. The kinase-coding region also coincides with the conserved protein sequences which are found in other tyrosine kinases. A compact domain of the v-abl protein including this kinase-coding region can accumulate to high levels in bacteria. The C-terminal region of the v-abl protein is not needed for the kinase activity and is rapidly degraded in bacteria. PMID- 2981222 TI - Expression of the gene for Bacillus subtilis aspartokinase II in Escherichia coli. AB - The gene coding for the subunits of aspartokinase II from Bacillus subtilis has been identified in a B. subtilis DNA library and cloned in a bacterial plasmid (Bondaryk, R. P., and Paulus, H. (1984) J. Biol. Chem. 259, 585-591). The introduction of a plasmid carrying the aspartokinase II gene into an auxotrophic Escherichia coli strain lacking all three aspartokinases restored its ability to grow in the absence of L-lysine, L-threonine, and L-methionine. The B. subtilis aspartokinase gene could thus be functionally expressed in E. coli and substitute for the E. coli aspartokinases. Measurement of aspartokinase levels in extracts of aspartokinaseless E. coli transformed with the B. subtilis aspartokinase II gene revealed an enzyme level comparable to that in a genetically derepressed B. subtilis strain. In spite of the high level of aspartokinase, the growth of the transformed E. coli strain was severely inhibited by the addition of L-lysine but could be restored by also adding L-homoserine. This apparently paradoxical sensitivity to lysine was due to the allosteric inhibition of B. subtilis aspartokinase II by that amino acid, a property which was also observed in extracts of the transformed E. coli strain. The synthesis and degradation of the aspartokinase II subunits were measured by labeling experiments in E. coli transformed with the B. subtilis aspartokinase II gene. In contrast to exponentially growing cells of B. subtilis which contained equimolar amounts of the aspartokinase alpha and beta subunits, the transformed E. coli strain contained a 3-fold molar excess of beta subunit. Pulse-chase experiments showed that the disproportionate level of beta subunit was not due to more rapid turnover of alpha subunit, both subunits being quite stable, but presumably to a more rapid rate of synthesis. After the addition of rifampicin, the synthesis of alpha subunit declined much more rapidly than that of beta subunit, indicating that the two subunits were translated independently from mRNA species that differ in functional stability. In conjunction with the results described in the preceding paper which demonstrated that the aspartokinase subunits are encoded by a single DNA sequence, these observations imply that the alpha and beta subunits of B. subtilis aspartokinase II are the products of in-phase overlapping genes. PMID- 2981224 TI - Characterization of a cyanobacterial photosystem I complex. AB - A simple procedure is described for the preparation of photosystem I (PSI) particles from Triton X-100-solubilized thylakoid membranes of the unicellular cyanobacterium Synechococcus 6301. The purified PSI complex contained the full complement of antenna chlorophylls, 130 +/- 5/P700, displayed the electron paramagnetic resonance signals characteristic of iron-sulfur centers X, A, and B, and had a protein/chlorophyll ratio of 2.9. Determination of the polypeptide composition, utilizing a uniformly 14C-labeled complex, showed that it contained polypeptides of 70, 18, 17.7, 16, and 10 kDa, in a molar ratio of 4.0:0.7:1.0:0.5:1.6. The relative amount of the lower molecular weight polypeptides showed progressive decrease with increase in Triton X-100 concentration and time of exposure to detergent. Consequently, it is proposed that in vivo the composition of the complex is [70 kDa]4 [18 kDa]1 [17.7 kDa]1 [16 kDa]1 [10 kDa]2. Relative to 130 mol of chlorophyll a, the PSI complex contained 16 mol of carotenoids, 13.7 +/- 1.0 g atoms of Fe, and 12.2 +/- 1.1 g atoms of labile sulfide. The properties of complexes fully depleted of the low molecular weight polypeptides by treatment with sodium dodecyl sulfate or with proteinase K are also described. PMID- 2981225 TI - Molecular characteristics of nerve growth factor receptors on PC12 cells. AB - Cross-linking of 125I-nerve growth factor (NGF) to PC12 cells with the photoreactive heterobifunctional agent N-hydroxysuccinimidyl-4-azidobenzoate results in the labeling of two major bands with Mr 158,000 and 100,000 and a minor band with Mr 225,000 as determined by polyacrylamide gel electrophoresis under denaturing and reducing conditions. Binding of 125I-NGF to and cross linking into all these species is abolished in the presence of excess unlabeled NGF but not in the presence of unlabeled epidermal growth factor, insulin, or bovine pancreatic trypsin inhibitor. When PC12 cells with bound 125I-NGF are incubated in excess unlabeled NGF at 0 degree C prior to cross-linking, only the Mr 158,000 species remains. In addition, binding of 125I-NGF to the Mr 158,000 complex is trypsin-resistant, whereas binding to the Mr 100,000 complex is not. These experiments identify the Mr 158,000 species as the slow NGF-receptor complex (chase stable at 0 degree C) and the smaller Mr 100,000 species as the fast NGF-receptor complex (trypsin sensitive). Furthermore, 125I-NGF bound to the former but not to the latter species is displaced by very-low concentrations of NGF, showing that at least a significant fraction of the high-molecular-weight slow receptor is also a high-affinity receptor. This identification is supported by the finding that chick sensory neurons which possess both high- and low affinity receptors exhibit two major labeled bands with Mr 145,000 and 105,000 as a result of cross-linking with 125I-NGF, whereas a cell population enriched in non-neuronal cells, which possess only low-affinity receptors, exhibits only the Mr 105,000 component. A shift in molecular weight of both species after pretreatment with neuraminidase indicates that both complexes contain sialoglycoproteins and rules out the possibility that differences in sialic acid content are responsible for the difference in molecular weight of the two complexes. The relative amount of the labeling of these two complexes is not affected by the presence of protease inhibitors nor by a variation of 5000-fold in cross-linker concentration. These results place some limits on possible models for the NGF receptors and their interconversion. PMID- 2981226 TI - Identification of multiple sites in the promoter region of the Tetrahymena pyriformis rRNA gene which bind the Escherichia coli catabolite regulatory protein. AB - The base sequence of the Tetrahymena pyriformis rRNA gene transcription initiation region has been reported (Niles, E. G., Sutiphong, J., and Haque, S. (1981) J. Biol. Chem. 256, 12849-12856). Upstream from the transcription initiation point, a collection of three families of highly conserved repeated sequences were identified, which may play a role in the expression of this gene. One of these families, repeat II, which has 10 members repeated in tandem, was recently observed to be strikingly similar in sequence to the Escherichia coli catabolite regulatory protein recognition site. In order to determine if these sequences can serve as catabolite regulatory protein binding sites, direct binding experiments were undertaken using a combination of the gel electrophoretic mobility alteration assay (Fried, M. G., and Crothers, D. M. (1983) Nucleic Acids Res. 11, 141-158) and DNA footprinting. DNA sequence specific and cyclic AMP-dependent binding of catabolite regulatory protein to several of the apparent recognition sites has been demonstrated. PMID- 2981227 TI - Ribonucleotide reductase activity and deoxyribonucleoside triphosphate metabolism during the cell cycle of S49 wild-type and mutant mouse T-lymphoma cells. AB - We investigated deoxyribonucleoside triphosphate metabolism in S49 mouse T lymphoma cells synchronized in different phases of the cell cycle. S49 wild-type cultures enriched for G1 phase cells by exposure to dibutyryl cyclic AMP (Bt2cAMP) for 24 h had lower dCTP and dTTP pools but equivalent or increased pools of dATP and dGTP when compared with exponentially growing wild-type cells. Release from Bt2cAMP arrest resulted in a maximum enrichment of S phase occurring 24 h after removal of the Bt2cAMP, and was accompanied by an increase in dCTP and dTTP levels that persisted in colcemid-treated (G2/M phase enriched) cultures. Ribonucleotide reductase activity in permeabilized cells was low in G1 arrested cells, increased in S phase enriched cultures and further increased in G2/M enriched cultures. In cell lines heterozygous for mutations in the allosteric binding sites on the M1 subunit of ribonucleotide reductase, the deoxyribonucleotide pools in S phase enriched cultures were larger than in wild type S49 cells, suggesting that feedback inhibition of ribonucleotide reductase is an important mechanism limiting the size of deoxyribonucleoside triphosphate pools. The M1 and M2 subunits of ribonucleotide reductase from wild-type S49 cells were identified on two-dimensional polyacrylamide gels, but showed no significant change in intensity during the cell cycle. These data are consistent with allosteric inhibition of ribonucleotide reductase during the G1 phase of the cycle and release of this inhibition during S phase. They suggest that the increase in ribonucleotide reductase activity observed in permeabilized S phase enriched cultures may not be the result of increased synthesis of either the M1 or M2 subunit of the enzyme. PMID- 2981228 TI - Effects of thyroid status on membrane-bound low Km cyclic nucleotide phosphodiesterase activities in rat adipocytes. AB - Adipocyte membranes from hypothyroid rats showed increased low Km cAMP phosphodiesterase activity compared to normals, provided that the subcellular fractionations were done in isotonic, as opposed to hypotonic, buffers. The enhanced cAMP phosphodiesterase activity in hypothyroid membranes was nearly normalized by incubation with a 10-fold excess of cGMP. Preincubation of hypothyroid adipocytes with cGMP also restored to normal the blunted lipolytic response to micromolar concentrations of epinephrine. DEAE-Sephacel chromatography of detergent-solubilized membrane-bound cAMP phosphodiesterase showed a 2.5-fold enhancement in hypothyroid membranes of a form of the enzyme that was completely inhibited by cGMP; the enzymatic elution profiles of the soluble fractions showed no difference between normal and hypothyroid fat pads. The results suggest a possible regulatory role of cGMP in adipocytes in the hypothyroid state. PMID- 2981229 TI - Polarized delivery of viral glycoproteins to the apical and basolateral plasma membranes of Madin-Darby canine kidney cells infected with temperature-sensitive viruses. AB - The intracellular route followed by viral envelope glycoproteins in polarized Madin-Darby canine kidney cells was studied by using temperature-sensitive mutants of vesicular stomatitis virus (VSV) and influenza, in which, at the nonpermissive temperature (39.5 degrees C), the newly synthesized glycoproteins (G proteins) and hemagglutinin (HA), respectively, are not transported out of the endoplasmic reticulum. After infection with VSV and incubation at 39.5 degrees C for 4-5 h, synchronous transfer of G protein to the plasma membrane was initiated by shifting to the permissive temperature (32.5 degrees C). Immunoelectron microscopy showed that under these conditions the protein moved to the Golgi apparatus and from there directly to a region of the lateral plasma membrane near this organelle. G protein then seemed to diffuse progressively to basal regions of the cell surface and, only after it had accumulated in the basolateral domain, it began to appear on the apical surface near the intercellular junctions. The results of these experiments indicate that the VSV G protein must be sorted before its arrival at the cell surface, and suggest that passage to the apical domain occurs only late in infection when tight junctions are no longer an effective barrier. In complementary experiments, using the temperature-sensitive mutant of influenza, cultures were first shifted from the nonpermissive temperature (39.5 degrees C) to 18.5 degrees C, to allow entrance of the glycoprotein into the Golgi apparatus (see Matlin, K.S., and K. Simons, 1983, Cell, 34:233-243). Under these conditions HA accumulated in Golgi stacks and vesicles but did not reach the plasma membrane. When the temperature was subsequently shifted to 32.5 degrees C, HA rapidly appeared in discrete regions of the apical surface near, and often directly above, the Golgi elements, and later diffused throughout this surface. To ensure that the anti-HA antibodies had access to lateral domains, monolayers were treated with a hypertonic medium to dilate the intercellular spaces. Some labeling was then observed in the lateral plasma membranes soon after the shift, but this never increased beyond 1.0 gold particle/micron, whereas characteristic densities of labeling in apical surfaces soon became much higher (approximately 10 particles/micron). Our results suggest that the bulk of HA follows a direct pathway leading from the Golgi to regions of the apical surface close to trans-Golgi cisternae. PMID- 2981230 TI - Tissue-specific expression of distinct spectrin and ankyrin transcripts in erythroid and nonerythroid cells. AB - cDNA probes for three components of the erythroid membrane skeleton, alpha spectrin, beta spectrin, and ankyrin, were obtained by using monospecific antibodies to screen a lambda gt11 expression vector library containing cDNA prepared from chicken erythroid poly(A)+ RNA. Each cDNA appears to hybridize to one gene type in the chicken genome. Qualitatively distinct RNA species in myogenic and erythroid cells are detected for beta spectrin and ankyrin, while alpha spectrin exists as a single species of transcript in all tissues examined. This tissue-specific expression of RNAs is regulated quantitatively during myogenesis in vitro, since all three accumulate only upon myoblast fusion. Furthermore, RNAs for two of the three genes do not accumulate to detectable levels in chicken embryo fibroblasts, demonstrating that their accumulation can be noncoordinate. These observations suggest that independent gene regulation and tissue-specific production of heterogeneous transcripts from the beta spectrin and ankyrin genes underlie the formation of distinct membrane skeletons in erythroid and muscle cells. PMID- 2981231 TI - Analytical study of microsomes and isolated subcellular membranes from rat liver. IX. Nicotinamide adenine dinucleotide glycohydrolase: a plasma membrane enzyme prominently found in Kupffer cells. AB - The distribution of nicotinamide adenine dinucleotide (NAD) glycohydrolase in rat liver was investigated by subcellular fractionation and by isolation of hepatocytes and sinusoidal cells. The behavior of NAD glycohydrolase in subcellular fractionation was peculiar because, although the enzyme was mainly microsomal, plasma membrane preparations contained distinctly more NAD glycohydrolase than could be accounted for by their content in elements derived from the endoplasmic reticulum or the Golgi complex identified by glucose-6 phosphatase and galactosyltransferase, respectively. When microsomal and plasmalemmal preparations were brought to equilibrium in a linear-density gradient, NAD glycohydrolase differed from these enzymes and behaved like 5' nucleotidase and alkaline phosphodiesterase I. NAD glycohydrolase was markedly displaced towards higher densities after treatment with digitonin. This behavior in density-gradient centrifugation strongly suggests that NAD glycohydrolase is an exclusive enzyme of the plasma membrane. NAD glycohydrolase differed clearly from other plasmalemmal enzymes when the liver was fractionated into hepatocytes and sinusoidal cells; its specific activity was considerably greater in sinusoidal cell than in hepatocyte preparations. Further subfractionation of sinusoidal cell preparations into endothelial and Kupffer cells by counterflow elutriation showed that NAD glycohydrolase is more active in Kupffer cells. We estimate that the specific activity of NAD glycohydrolase activity is at least 65 fold higher at the periphery of Kupffer cells than at the periphery of hepatocytes. As the enzyme shows not structure-linked latency and is an exclusive constituent of the plasma membranes, we conclude that it is an ectoenzyme that cannot lead to a rapid turnover of the cytosolic pyridine nucleotides. PMID- 2981232 TI - Cyclic AMP effects on cell-to-cell junctional membrane permeability during adipocyte differentiation of 3T3-L1 fibroblasts. AB - Mouse 3T3-L1 fibroblast cells, also know as preadipocytes, differentiate in vitro into adipocytes when treated with promoting agents and acquire numerous properties characteristic of mature fat cells. We studied junctional cell-to-cell communication by measuring the incidence of electrical coupling and transfer of carboxy-fluorescein among these cells. When 3T3-L1 cells were induced to differentiate into adipocytes, they lost virtually all cell-cell communication. Preadipocytes that remained nondifferentiated after the treatment maintained normal communication. Loss of communication in the adipocytes invariably coincided with appearance of lipid droplets and not with other phenotypic changes. In the differentiating cells, loss of cell-to-cell communication and lipid accumulation was prevented if dibutyryl cyclic AMP and caffeine were present in the culture medium. Addition of dibutyryl cyclic AMP and caffeine to already differentiated adipocytes resulted in loss of lipid and simultaneously improved junctional permeability. The results demonstrate that in the in vitro 3T3-L1 cell system, (a) cell-to-cell communication and lipid synthesis are intimately related during the adipose conversion and (b) cAMP affects the expression of the two phenotypes. PMID- 2981234 TI - Correlation between trypsin binding to a specific receptor and prostacyclin production in cultured vascular endothelial cells. AB - The correlation between the binding and processing of trypsin and its effect on prostacyclin (PGI2) production in cultured adult bovine aortic endothelial (ABAE) cells was studied. ABAE cells demonstrated an ability to produce PGI2 in a dose response manner to trypsin at the range of 0.1-2.0 micrograms/ml with a saturation at a concentration of 1 microgram/ml. Likewise, 125I-trypsin binding to the cultured cells increased in a dose-response way and reached saturation at a concentration of about 1 microgram/ml; 125I-trypsin was bound to a specific high-affinity cell-surface receptor with a dissociation constant (Kd) of 1.5 X 10(-8) M and each of the confluent ABAE cells has about 1.2 X 10(5) such receptors sites. The cell-surface receptor for trypsin displayed specific characteristics and an excess amount of unlabeled trypsin successfully abolished 125I-trypsin binding while thrombin in excess failed to compete for 125I-trypsin binding. Only a small fraction of the cell-surface-bound 125I-trypsin was internalized and subsequently degraded by ABAE cells as compared to the process of 125I-trypsin internalization by human skin fibroblasts (HSF). This study demonstrated that the stimulatory effect of trypsin on prostacyclin production and release by ABAE cells might be mediated by a specific cell-surface receptor for trypsin on these cells distinct from the thrombin receptor. PMID- 2981235 TI - EGF induces receptor down-regulation with no receptor recycling in KB cells. AB - Several ligands, including epidermal growth factor (EGF), have been found to negatively modulate or down-regulate their specific plasma membrane receptors. Using both 125I-EGF and a monoclonal antibody against the EGF-receptor (EGF-R1), we studied the down-regulation of the EGF-receptor in the human adenocarcinoma cell line KB. The results presented here demonstrate that incubating KB cells at 37 degrees C with EGF rapidly decreases the number of plasma membrane EGF receptors. In addition, there is a concomitant rise of equal magnitude in the number of EGF molecules taken up. The latter result argues strongly that there is negligible recycling of the EGF-receptor in KB cells and that the major portion of internalized EGF-receptor complexes are transported to lysosomes and subsequently degraded. The fate of the EGF-receptor is markedly different from that of receptors not subject to down-regulation. The biochemical signals that operate to regulate such diverse receptor traffic in cells remains to be elucidated. PMID- 2981236 TI - Regulation of angiotensin I-converting enzyme activity in serially cultivated bovine endothelial cells. AB - Angiotensin I-converting enzyme (ACE) activity was measured in lysates of cloned and uncloned cultures of bovine fetal aortic endothelial cells. The expression of ACE activity in these cells was complex, and influenced by subcultivation, cell density, serum, cumulative population doublings, and clonal heterogeneity. The ACE specific activity at any point in the in vitro lifespan was determined, at least in part, by interaction of these culture variables. After subcultivation to subconfluent densities, cellular ACE specific activity decreased markedly and did not reach detectable levels until cells attained confluent densities. The use of different suppliers' lots of serum in the growth medium resulted in different cellular ACE specific activities. The ACE specific activity decreased as cultures were serially subcultivated, but remained detectable throughout the lifespan, suggesting a linkage between the proliferative history of an endothelial cell and its remaining capacity to express ACE. Increased ACE activity was observed when cells at the end of their lifespan were cultured at high densities. Cloned strains behaved similarly to the uncloned parent culture, except that they exhibited a wide range of ACE specific activities. PMID- 2981233 TI - Synthesis of the transferrin receptor by cultures of embryonic chicken spinal neurons. AB - We have purified a glycoprotein from chicken sciatic nerves, sciatin, which has pronounced trophic effects on avian skeletal muscle cells in culture. Recent studies have shown that sciatin is identical to the iron-transport protein, transferrin, in terms of its physicochemical structure, immunological reactivity, and biological activity. To determine whether transferrin is synthesized and released by neuronal tissue, we incubated cultures of dissociated chicken spinal neurons in a medium free of L-leucine containing either L-3H-amino acids or L [14C]leucine and immunoprecipitated transferrin with highly specific antibodies. The radiolabeled protein precipitated by rabbit heteroclonal, goat heteroclonal, or mouse monoclonal antitransferrin antibodies increased in specific activity in a linear manner for at least 30 min. Synthesis of this protein was abolished by the presence of puromycin (20 micrograms/ml) or cycloheximide (10(-5) M). The disappearance of the radiolabeled protein from cells was linear with a half-life (t 1/2) of 8-10 h. When immunoprecipitates were separated by SDS gel electrophoresis, a prominent band corresponding to transferrin (Mr 84,000) was visualized by staining with Coomassie Blue. However, when such gels were fluorographed, no radioactivity was apparent in the transferrin region of the gel although a prominent radioactive band was visualized at an Mr of 56,000. The protein of Mr 56,000 was not simply a degradation product of transferrin because this particular protein band was not generated by incubating radiolabeled transferrin with unlabeled neuronal homogenates. The protein of Mr 56,000 was purified from embryonic chicken brain and spinal cord by immunoabsorption chromatography on mouse monoclonal antitransferrin IgG conjugated to Sepharose 4B followed by affinity chromatography on immobilized transferrin. The purified protein bound radioiodinated transferrin and was precipitated by rabbit anti chicken transferrin-receptor antibodies. Furthermore, this receptor protein was found to be localized on the plasma membrane of dorsal root ganglion neurons by immunocytochemistry using the peroxidase-antiperoxidase technique, and by blocking experiments, which showed that antitransferrin receptor IgG could inhibit the binding of fluorescein-conjugated transferrin at 4 degrees C to cultured neurons in vitro. From these data, we conclude that transferrin is not synthesized by cultures of chicken spinal cord neurons, but that the receptor for transferrin is synthesized by these cultures and is precipitated by antitransferrin antibodies as an antigen-receptor complex. PMID- 2981237 TI - Transient and sustained effects of hormones and calcium on membrane potential in a bone cell clone. AB - Measurements were made of the electrophysiological and cAMP response to changes in extracellular [Ca2+] and to hormone application in a bone cell clone. Both transient and long-term electrophysiological responses were studied. An increase in extracellular [Ca2+] usually resulted in a transient hyperpolarization of about 60-sec duration. In addition, increases in extracellular [Ca2+] from 0.9 to 1.8 mM and from 1.8 to 3.6 mM resulted in long-term hyperpolarization and increased potential fluctuations. Increasing bathing [Ca2+] until the membrane potential reached the K+ equilibrium level resulted in a significant decrease in fluctuations. Addition to the bathing medium of quinine, a putative blocker of the Ca2+-dependent K+ channel, resulted in long-term depolarization of the mean membrane potential, and a long-term decrease in potential fluctuations. Addition of Mg2+, a mild antagonist of Ca2+ entry into the cell, produced transient depolarization and reduction of potential fluctuations. These effects suggest that the potential fluctuations reflect cytoplasmic [Ca2+] fluctuations via Ca2+ dependent K+ membrane channels. Under an extracellular [Ca2+] of 1.8 mM, the application of prostaglandin E2 (PGE2), isoproterenol, and parathyroid hormone produced no significant effect on mean membrane potential or on the sustained potential fluctuations, but PGE2 did significantly raise intracellular cAMP. Under an increased bathing [Ca2+], significant changes in mean potential and fluctuations did occur in response to PGE2, but not in response to the other hormones, while the PGE2 effect on cAMP was not greatly changed. Hyperpolarizing transients of about 30-sec duration occurred in response to all of the hormones, particularly at an extracellular [Ca2+] of 3.6 mM. Thus, there are both transient and long-term electrophysiological responses to hormone application, with only the long-term response correlated with the production of cAMP. These electrophysiological responses may represent separate transient and long-term calcium transport responses to hormone application. PMID- 2981238 TI - Cultured mouse marrow stromal cell lines. II. Distinct subtypes differing in morphology, collagen types, myelopoietic factors, and leukemic cell growth modulating activities. AB - A series of stromal cell lines were studied for their growth properties, electron microscopic morphology, cytochemical profile, collagen types, production of myelopoietic factors, and modulation of leukemic cell growth. Three cell types were identified in addition to the previously described macrophages (14M and 14M1) and preadipocytes (14F). MBA-1 cells were found to be fibroblasts by their ability to synthesize collagen types I and III, while the cell line MBA-13 shared properties in common with both fibroblasts and endothelial cells (collagen types I, III, IV, V). The third cell type, represented by the stromal cell line MBA-2, produced mainly collagen types IV and V and exhibited junctional complexes between adjacent cells. All of the cell lines tested produced and secreted a macrophage-colony-stimulating factor, CSF-1. MBA-2 and to a lesser extent, MBA 13, produced an additional activity resistant to anti-CSF-1 antiserum. Trypsin extraction of outer surface components from two clones of the MBA-2 cell line (MBA-2.1 and MBA-2.4) yielded high molecular weight factor(s) that specifically inhibited the growth of a plasmacytoma cell line (MPC-11). Such inhibitory activity was not detected in other stromal cell lines. It is possible that this variability in the nature of stromal cell lines represents corresponding diversity of cell types comprising the hematopoietic microenvironment in vivo. PMID- 2981239 TI - A comatose alcoholic. PMID- 2981240 TI - The constant plasma 18-hydroxycorticosterone to aldosterone ratio: an expression of the efficacy of corticosterone methyloxidase type II activity in disorders with variable aldosterone production. AB - Aldosterone and 18-hydroxycorticosterone (18-OHB) are produced by the adrenocortical zona glomerulosa. Under normal conditions, plasma 18-OHB levels parallel and are influenced by the same trophic factors that regulate aldosterone production. To evaluate corticosterone-methyl-oxidase II activity, the final step of aldosterone biosynthesis, in conditions associated with chronic derangements of the pituitary-adrenal and/or renal-adrenal axis, we measured the plasma 18-OHB to aldosterone ratio, cortisol, PRA or plasma renin concentration, and potassium (K) in 104 such patients and 15 normal subjects. The 18-OHB to aldosterone ratios in the pituitary-adrenal group were not significantly different from normal regardless of elevated or reduced ACTH and/or cortisol levels [patients with Cushing's syndrome, 3.55 +/- 0.68 (+/-SE); ACTH deficiency, 2.03 +/- 0.34; 21 hydroxylase deficiency, 3.09 +/- 0.23; normal subjects, 2.50 +/- 0.15]. The renal adrenal group also had normal ratios regardless of plasma renin concentration and K levels [patients with aldosterone-producing adenomas, 2.85 +/- 0.15; idiopathic hyperaldosteronism, 2.14 +/- 0.19; salt-losing nephropathy, 3.06 +/- 0.54; Bartter's syndrome, 2.89 +/- 0.20; isolated (hyporeninemic) hypoaldosteronism, 3.20 +/- 0.39]. Only in patients with 17 alpha-hydroxylase deficiency (230.1 +/- 118.6) was the ratio abnormally high. Chronic perturbations of aldosterone production by ACTH, the renin-angiotensin system, and/or K did not alter this last step of aldosterone biosynthesis (corticosterone-methyloxidase II), as defined by their plasma concentrations. Any influence of these trophic factors must be proximal to the site of 18-OHB production. PMID- 2981241 TI - Glucocorticoid receptors and responsiveness of normal and neoplastic human adrenal cortex. AB - Glucocorticoids have been postulated to directly inhibit adrenocortical steroid production in laboratory animals. To investigate this in the human, we measured specific [3H]dexamethasone-binding sites in cytosol samples from normal and neoplastic human adrenal tissues. All nine normal adrenocortical samples, six adenomas (four cortisol-producing and two aldosterone-producing), and two hyperplastic adrenocortical samples studied were devoid of measurable specific glucocorticoid-binding activity. In contrast, steroid binding with characteristics of the glucocorticoid receptor (concentration of binding sites, 32-146 fmol/mg cytosol protein; Kd, 1.7-3.1 X 10(-9) M) was readily detectable in cytosol of all three adrenocortical carcinomas and all three pheochromocytomas examined. To elucidate the in vivo role of glucocorticoids as direct regulators of adrenocortical function, five patients with hypopituitarism receiving varying oral maintenance doses of dexamethasone were given ACTH iv. Increasing the orally administered dexamethasone dose from 1 to 8 mg/day did not alter the plasma cortisol response to a 4-h infusion of 250 micrograms synthetic ACTH in these patients. Collectively, these data cast doubt on the proposal that synthetic glucocorticoids directly suppress adrenocortical function in the human. Whether glucocorticoid receptors in tumor tissue could mediate the dexamethasone-induced suppression of hypercortisolism occasionally reported in patients with adrenocortical neoplasia remains to be investigated. PMID- 2981242 TI - Opioid effects on plasma concentrations of luteinizing hormone and prolactin in the adult male rhesus monkey. AB - The role of endogenous opioid peptides (EOP) in the neuroendocrine control of primate gonadotropin and PRL secretion was studied in nonrestrained adult male rhesus monkeys. Morphine (0.5-1.0 mg/kg) was used as the prototype opiate, beta endorphin (beta-END; 10-20 micrograms/kg) and [D-Ala2,D-Leu5] enkephalin (DADLE; 5-20 micrograms/kg) were used as representatives of EOP, and naloxone (0.5-2.0 mg/kg) was used as an opiate receptor blocker. Drugs were administered and blood was collected (at 20-min intervals for 4 h) through an indwelling jugular catheter. LH and PRL levels were measured in plasma by RIA. Intravenous administration of morphine (1.0 mg/kg) and DADLE (10 micrograms/kg) produced decreases in LH levels of 64% and 40%, respectively. These decreases occurred within 1 h after drug injections and lasted for approximately 3 h. beta-END had no effect on LH levels. Naloxone, at all doses studied, significantly increased LH levels (5- to 8-fold). The LH rises occurred within 20 min and lasted for up to 2 h. Both morphine and beta-END produced immediate increases in PRL, which remained elevated for 3 h. DADLE did not alter PRL levels. Naloxone (1.0 and 2.0 mg/kg) decreased PRL concentrations (45% and 60%, respectively). Pretreatment with morphine or DADLE did not alter the LH response to GnRH (100 micrograms) stimulation, indicating a hypothalamic site of action for the opioid inhibition of LH release. Naloxone administration reversed the inhibitory effects of morphine and DADLE on LH. The stimulatory effect of morphine on PRL levels was also reversed by naloxone. These studies further define the postulated physiological role of EOP in primate reproductive neuroendocrinology. Based on receptor selectivities of these opioid agonists, the inhibition of LH may be mediated by delta-receptors, whereas PRL release appears to be mu-mediated. PMID- 2981243 TI - Plasma beta-endorphin levels in primary aldosteronism. AB - Excessive production of an as yet unidentified aldosterone-stimulating factor may cause idiopathic hyperaldosteronism (IHA). This putative factor may be related to proopiomelanocortin-derived peptides, some of which have aldosterone-stimulating properties. The present study evaluated plasma beta-endorphin, ACTH, cortisol, and aldosterone levels in patients with IHA (n = 10), aldosterone-producing adenomas (n = 4), essential hypertension (n = 11), and normal subjects (n = 10). Plasma and urinary hormone measurements were obtained at timed intervals during an isocaloric, fixed electrolyte intake (Na+, 128 meq/day; K+, 80 meq/day) in a metabolic unit. Plasma for beta-endorphin assay was preincubated with sepharose bound anti-beta-lipotropin to remove beta-lipotropin that cross-reacted with the beta-endorphin RIA. Mean +/- SE plasma beta-endorphin levels at 0800 h were elevated in IHA patients (47 +/- 13 fmol/ml) compared to those in aldosterone producing adenoma (25 +/- 9), essential hypertension (16 +/- 1), and normal control (20 +/- 2; P less than 0.05) subjects. Plasma ACTH, plasma cortisol, and urinary cortisol levels were not different in these four groups. These data support the hypothesis that excess production of either beta-endorphin or related proopiomelanocortin-derived peptides may function as aldosterone secretogogue(s) in IHA. PMID- 2981244 TI - Cyclical Cushing's disease: two distinct rhythms in a patient with a basophil adenoma. AB - A 71-yr-old woman with clinical signs of Cushing's syndrome was studied continuously for an extended period after demonstration of a paradoxical response to dexamethasone. She proved to have a corticotroph cell adenoma of the pituitary which caused secretion of ACTH and cortisol in two distinct rhythms. One rhythm consisted of a period of 40 days of excess cortisol production, followed by a period of 60-70 days of normal production. During the period of excess cortisol production there was a second rhythm, consisting of peaks of cortisol production every 3-6 days with intervening troughs of normal cortisol production. Prolonged clinical remission followed transphenoidal surgery, but the pituitary still has the ability to provoke abnormal amounts of cortisol secretion, as occurred during a postoperative dexamethasone suppression test. The long duration of normal cortisol production phases in this patient demonstrates the difficulty in excluding Cushing's syndrome in patients with suggestive clinical symptoms but normal serum and urinary cortisol levels if these tests are measured for a single short phase of several days. PMID- 2981245 TI - Insulin-stimulated receptor phosphorylation appears normal in cultured Epstein Barr virus-transformed lymphocyte cell lines derived from patients with extreme insulin resistance. AB - Insulin-stimulated phosphorylation of the insulin receptor was studied in cultured B-lymphocytes transformed by Epstein-Barr virus. In studies with cell lines derived from six normal subjects, insulin (10(-7) M) caused an average increase of approximately 200% in 32P incorporation into the 95K subunit of the insulin receptor. Phosphorylation was rapid (detectable within 1-2 min) and reached a maximum level by 15 min. Dose-response curves for receptor occupancy and phosphorylation were nearly superimposable, indicating few or absent spare receptors for this response to insulin. These data suggest that insulin receptor phosphorylation is an early response to insulin in cultured lymphocytes transformed with Epstein-Barr virus. We studied insulin receptor phosphorylation in cell lines derived from nine patients with clinical syndromes associated with extreme insulin resistance, all of whom had normal [125I] insulin binding. While the magnitude of insulin's stimulation varied widely among the individual cell lines, no significant differences were found between cell lines from normal subjects and those from patients with extreme insulin resistance. PMID- 2981246 TI - Characterization of insulin-like growth factor II binding to human fibroblast monolayer cultures. AB - Two major somatomedin peptides have been isolated from human plasma, somatomedin C/insulin-like growth factor I (SMC/IGF-I) and insulin-like growth factor II (IGF II). Also, two types of SM/IGF receptors have been defined. Type I receptors have a higher affinity for SMC/IGF-I than IGF-II, and insulin binds to this receptor at high concentrations. Type II receptors have a higher affinity for IGF-II than SMC/IGF-I, and insulin does not bind to this receptor site. In this study, we characterized the binding of IGF-II to human monolayer fibroblast cultures, and the affinity and specificity of this binding. We also compared the binding of IGF II to the binding of insulin and SMC/IGF-I to these cells. The receptors for IGF II on normal human fibroblast monolayers fit the criteria for type II SM/IGF receptors, and there were more type II receptors on these cells than either insulin receptors or type I SM/IGF receptors. The type II receptors on human fibroblasts did not demonstrate autoregulation by homologous hormone, unlike the type I SM/IGF and insulin receptors. In addition, they were not changed by acute or chronic exposure to insulin. It is so far unclear what biological function IGF II plays in vivo. This human fibroblast system will be a valuable experimental model for the study of IGF-II receptors and their relationship to the biological actions of IGF-II. PMID- 2981247 TI - Macrophage response to herpes simplex encephalitis in immune competent and T cell deficient mice. AB - Corneal inoculation of Nude (athymic) mice and Balb/c mice with herpes simplex virus Type I produces a brainstem encephalitis with demyelination of the trigeminal root entry zone. The extent of CNS demyelination is less in the immune deficient athymic mice 7 days after infection compared to the immune-competent Balb/c mice. Both groups demonstrate a macrophage response and beginning myelin disruption approximately 3 days after corneal infection when herpes viral particles are first observed within central nervous system cells. Five to seven days after infection when differences in the extent of demyelination between the immune-competent and immune-deficient animals become evident, the Balb/c mice demonstrate T cells and increasing numbers of macrophages at the trigeminal root entry zones. These findings suggest an interaction between macrophages and T cells which leads to an extension of the demyelination in the immune competent Balb/c mice and that lack of T cells is important in limiting demyelination in Nude (athymic) mice. PMID- 2981248 TI - Feline glucose-6-phosphate dehydrogenase cellular mosaicism. Application to the study of retrovirus-induced pure red cell aplasia. AB - Neoplasms result from the uncontrolled proliferation of abnormal or transformed cells. The early stages of this process are difficult to study because of the lack of sensitive and specific markers of clonal evolution in an experimental system. We have developed a cat model using cellular mosaicism for glucose-6 phosphate dehydrogenase (G-6-PD). Our findings confirm that the structural locus for feline G-6-PD is on the X-chromosome and demonstrate that it is randomly inactivated in somatic cells. Heterozygous cats have balanced ratios of G-6-PD enzyme types in peripheral blood cells and hematopoietic progenitors that remain stable over time. In our initial studies, we used the model to analyze the events surrounding marrow failure experimentally induced by selected strains of feline leukemia virus (FeLV). Two G-6-PD heterozygous cats, one F1 male hybrid and one domestic cat were infected with FeLV (C or KT) and developed pure red cell aplasia (PRCA). Colonies arising from the more mature erythroid colony-forming cell were not detected in marrow culture of anemic animals although erythroid bursts persisted, suggesting that the differentiation of early erythroid progenitors (BFU-E) was inhibited in vivo. The ratio of G-6-PD types in hematopoietic progenitors and peripheral blood cells from the heterozygous cats did not change when the animals developed PRCA. Thus, the anemia did not result from the clonal expansion of a transformed myeloid stem cell. With this experimental approach, one may prospectively assess clonal evolution and cellular interactions in other FeLV-induced diseases. PMID- 2981249 TI - Asialo von Willebrand factor interactions with platelets. Interdependence of glycoproteins Ib and IIb/IIIa for binding and aggregation. AB - Asialo von Willebrand factor (AS-vWf) binds to and aggregates normal human platelets in the absence of ristocetin. Maximal specific binding of AS-vWf is 1-2 micrograms vWf protein/10(8) platelets. Despite the specificity of the binding, only 60% of the bound AS-vWf can be dissociated after equilibrium has been reached. We investigated the site of binding and the mechanism of aggregation of platelets by AS-vWf by (a) pre-incubating platelets with either of two monoclonal antibodies, one against glycoprotein Ib (GPIb) or a second against the glycoprotein IIb/IIIa complex (GPIIb/IIIa), and (b) varying the concentration of fibrinogen in the medium. The results of our studies indicate that AS-vWf binds initially to GPIb. This binding then results in the exposure of receptors for AS vWf on GPIIb/IIIa. In the presence of plasma fibrinogen, both AS-vWf and fibrinogen bind to GPIIb/IIIa. In the presence of plasma fibrinogen, 50% more AS vWf binds to the platelet, and this additional AS-vWf binds almost exclusively to GPIIb/IIIa. Despite this enhanced binding of AS-vWf in the absence of fibrinogen, platelet aggregation is much less than that which occurs in the presence of plasma fibrinogen. Comparative studies of AS-vWf binding to normal platelets and the platelets of patients with Glanzmann's thrombasthenia reveal decreased binding to the thrombasthenic platelets and a marked decrease in the extent of platelet aggregation. These studies indicate that AS-vWf binding to, and ensuing aggregation of, platelets is different from that observed with intact vWf protein when platelets are stimulated with either ristocetin or thrombin. The AS-vWf binds to GPIb which, in turn, makes additional AS-vWf receptors available on GPIIb/IIIa. If plasma fibrinogen is present, it competes with the AS-vWf for binding to GPIIb/IIIa and causes aggregation of platelets. In the presence of plasma fibrinogen, more of the AS-vWf binds to GPIIb/IIIa, but this AS-vWf is much less effective than fibrinogen in supporting platelet aggregation. PMID- 2981250 TI - Hydraulic water permeability and transepithelial voltage in the isolated perfused rabbit cortical collecting tubule following acute unilateral ureteral obstruction. AB - Ureteral obstruction affects the kidney's ability to conserve water and sodium. Using the isolated perfused tubule technique, we studied cortical collecting tubules (CCT) taken from rabbits subjected to a sham operation or to 4 h of unilateral ureteral obstruction (UUO). Tubules were perfused in the presence of an osmotic gradient directed to promote water movement from lumen to bath, and volume flux (Jv), hydraulic water permeability (Lp), and transepithelial voltage (V1) were determined. In tubules from sham-operated and UUO animals, basal (before exposure to vasopressin) J, and Lp were not different from zero. After addition of 200 microU . ml-1 of arginine vasopressin (aVP) to the bath, Jv and Lp increased to 1.64 +/- 0.23 nl . mm-1 . min-1 and 127.9 +/- 19.8 cm . s-1 . atm 1 x 10(7), respectively, in tubules from sham-operated animals, but not only 0.27 +/- 0.09 nl . mm-1 . min-1 an 18.8 +/- 6.2 cm . s-1 . atm-1 . 10(7) in tubules from UUO animals. Pretreatment with desoxycorticosterone acetate (DOCA) or indomethacin in vivo did not prevent the blunted vasopressin response seen in tubules taken from UUO animals. The Jv and Lp responses to the cyclic AMP (cAMP) analogue, 8-Br-cAMP, were also diminished in tubules taken from UUO animals compared with shams. V1, measured during the basal period, was diminished in tubules from UUO kidneys (-5.0 +/- 2.1 mV) compared with shams (-21.9 +/- 4.1 mV), and pretreatment with DOCA did no prevent the effects of UUO on V1. In contrast, tubules taken from animals that received indomethacin prior to UUO developed voltages not different from voltages in tubules taken from sham operated animals (-17.3 +/- 1.7 mV). We conclude that, although CCT from UUO animals can maintain osmotic gradients, their ability to respond to vasopressin by increasing Lp is impaired by an intrinsic defect located at a step beyond the generation of cAMP, and that prostaglandin inhibition or DOCA pretreatment do not reverse the decreased responsiveness of Lp to aVP. UUO also diminished V1, and this abnormality was prevented by previous treatment with indomethacin, suggesting that prostaglandins may mediate the effect of UUO on V1. PMID- 2981251 TI - Gastrin releasing peptide is a selective mitogen for small cell lung carcinoma in vitro. AB - Human small cell lung carcinoma (SCLC) cells have been shown to contain significant levels of a bombesin-immunoreactive peptide. The 27-amino acid peptide, gastrin releasing peptide (GRP), has recently been shown to be responsible for the bombesin-like immunoreactivity found in SCLC cells. Among four lung cancer cell lines examined in vitro, GRP exhibited mitogenic activity for two SCLC subtypes, but not for a squamous carcinoma or adenocarcinoma lung cell line. The mitogenicity of the GRP molecule has been isolated to the carboxyterminal fragment, designated GRP 14-27, which is in part homologous to bombesin. The aminoterminal fragment, GRP 1-16, is no homologous to bombesin and exhibits no mitogenic activity. Thus, GRP may be an important growth regulating or autocrine factor in human SCLC. PMID- 2981252 TI - Locus assignment of alpha-globin structural mutations by hybrid-selected translation. AB - The two human alpha-globin genes, alpha 1 and alpha 2 located 3.4 kilobases apart on chromosome 16, encode identical alpha-globin proteins. A mutation in either gene could result in a structural hemoglobinopathy. It has only recently become possible to assign an alpha-chain mutant to one of these two loci by using recombinant DNA technology. While definitive, this approach has necessitated the cloning and sequencing of the specific gene in question. We present an alternative approach which results in rapid and definitive assignment of an alpha globin mutation to its encoding genetic locus. This approach uses the technique of hybrid-selected translation. Reticulocyte RNA from individuals with alpha globin mutations can be fractionated into beta-, alpha 9 (total)-, alpha 1-, and alpha 2-globin mRNA by selective hybridization of each mRNA species to its respective complementary DNA (cDNA) immobilized on nitrocellulose paper. Each mRNA purified in this way can be translated in vitro, and the mRNA species (and hence gene locus) encoding the globin mutant can then be directly identified by gel analysis of the radiolabeled translation products. This procedure can be used to identify globin mutants as alpha or beta and to localize alpha-globin mutants to the alpha 1 or alpha 2 gene. We have used this technique to localize the two alpha-globin mutants, alpha 125Pro (Hb Quong Sze) and alpha 47HIS (Hb Hasharon), to the alpha 2 locus. This approach could potentially be expanded to serve as an alternative to peptide analysis for the initial characterization of all globin structural mutants. PMID- 2981254 TI - Origin of leukemic relapse after bone marrow transplantation detected by restriction fragment length polymorphism. AB - Bone marrow transplantation has become an accepted modality in the treatment of acute leukemia. With this therapy, it is possible to obtain long-term disease free survival. However, leukemia recurs occasionally. In most cases, leukemic relapse is of recipient origin. There have been several reports, though, of leukemia developing in donor cells. These cases have been limited to instances in which there is an easily identifiable chromosome difference or abnormality, usually a sex chromosome. In this paper we describe the use of restriction fragment-length polymorphism analysis to determine the origin of recurrent leukemia cells in which no identifying chromosome was present. We found that the leukemia had recurred in recipient cells. We also were able to demonstrate the presence of normal hemopoietic cells of donor origin. PMID- 2981253 TI - Identification of carriers of a variant plasma prealbumin (transthyretin) associated with familial amyloidotic polyneuropathy type I. AB - A method is described for detecting carriers of a variant plasma prealbumin that is associated with familial amyloidotic polyneuropathy (FAP) type I. It is based on the finding of an extra methionine in the variant prealbumin, at position 30 from the amino terminals. Since normal prealbumin has only one methionine (position 13), treatment with cyanogen bromide (CNBr), which cleaves only at methionines, results in two peptides. CNBr treatment of the variant prealbumin gives three peptides. The extra can then be detected in two ways: by HPLC using a reverse phase C18 column, and by sequential Edman degradation. Each method can detect as little as 1% variant prealbumin in isolated plasma prealbumin, and therefore, can identify carriers of the gene for the variant protein. Since FAP type I usually is not manifest until after the childbearing years, this method to identify carriers of the gene offers a new approach for genetic counseling of families with this disease. To date, kindreds with hereditary amyloidosis that could benefit from these studies include those with FAP type I of Swedish, Japanese, and Portuguese origins. PMID- 2981255 TI - Radionuclide angiocardiography in children. AB - Clinical use of radiotracer techniques in pediatric cardiology is increasing. Three basic methods being employed are first pass radionuclide angiocardiography, gated blood pool scanning and myocardial perfusion scintigraphy. These methods are rapid, safe and accurate. They are minimally invasive and result in a low radiation dose to the patient. The development of ultrashort-lived radionuclides (such as iridium-191m with a half-life of only 5 seconds), improvements in nuclear imaging instrumentation and numerical analysis should contribute to a further utilization of these methods. PMID- 2981256 TI - A functional approach to stroke management in elderly patients. PMID- 2981257 TI - Adrenal insufficiency (Addison's disease) in the elderly. PMID- 2981258 TI - Effect of lipoxygenase metabolites of arachidonic acid on proliferation of human T cells and T cell subsets. AB - The lipoxygenase products LTB4 and 15 HPETE have been reported to stimulate T suppressor cell function and also to inhibit [3H]thymidine incorporation into mitogen-stimulated T cells. This present report documents that although these compounds do indeed inhibit [3H]thymidine incorporation into unfractionated T cells, they significantly enhance [3H]thymidine incorporation into T cell preparation enriched for cells bearing the cytotoxic suppressor cell phenotype identified by the OKT8 monoclonal antibody. The mitogen response of T cells enriched for OKT4+ helper-inducer cells is inhibited in manner similar to the response of unfractionated T cells. Thus, LTB4 and 15 HPETE stimulate both the function and the proliferation of the cytotoxic-suppressor T cell subset. PMID- 2981259 TI - Inhibition of cytotoxic T lymphocyte-mediated lysis by ETYA: effect independent of arachidonic acid metabolism. AB - Cytotoxic T lymphocyte (CTL)-mediated lysis of target cells was inhibited by 5,8,11,14-eicosatetraynoic acid (ETYA) and other inhibitors of the lipoxygenase pathway at concentrations that inhibited arachidonic acid metabolism in mixed lymphocyte cultures. Inhibition was reversible and selective for the "lethal hit" stage in the CTL-target interaction. Studies to define CTL-specific arachidonic acid metabolites demonstrated that cloned CTL populations have little or no capacity to metabolize arachidonic acid. Therefore, inhibitor actions appear to be independent of the effects on CTL arachidonic acid metabolism. Alternative explanations for inhibitory effects are discussed. PMID- 2981260 TI - Stimulation of human neutrophilic granulocyte chemotaxis by monoclonal antibodies. AB - Two mouse monoclonal antibodies, L12.2 and S5.22, were developed that are specific for human neutrophilic granulocytes and produce a twofold to threefold stimulation of n-formyl-methionine-leucyl-phenylalanine (FMLP)-induced chemotaxis. Stimulation of chemotaxis by the antibodies is specific for FMLP and is concentration dependent. L12.2 appears to be more potent in stimulating chemotaxis and is isotypically distinct from S5.22. In addition, although L12.2 reacts only with mature peripheral blood granulocytes, S5.22 reacts with leukemic cells of both myeloid and monocytic origin and with immature granulocyte precursor cells. This suggests that L12.2 interacts with an antigen that appears late in the differentiation pathway, whereas S5.22 binds to an antigen that is present throughout the myeloid lineage. By means of the under-agarose and Boyden chamber techniques, L12.2, but not S5.22, by itself was also found to be a potent granulocyte chemoattractant. Cells in a gradient of L12.2 display polarized and oriented morphology. L12.2 alone, but not S5.22, also stimulates granulocyte phagocytosis and induces superoxide anion production. Neither L12.2 nor S5.22 affected the release of myeloperoxidase or lysozyme from granulocytes either alone or in combination with FMLP, C5a, or the tumor promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). These results suggest that L12.2 interacts with a single antigenic determinant on granulocytes that is involved in chemotaxis, phagocytosis, and superoxide anion release. PMID- 2981261 TI - Immunoradiometric assay to measure the in vitro penetration of sporozoites of malaria parasites into hepatoma cells. AB - We describe here an immunoradiometric assay to quantitate the in vitro invasion of hepatoma cells by sporozoites. The assay measures levels of circumsporozoite (CS) antigen that remain associated with the hepatoma cells after their incubation with the parasites. Several observations show that these measurements reflect internalized rather than extracellular antigen. For example, when incubations were performed with nonviable parasites (sonicated or heated), or in the presence of metabolic inhibitors, such as sodium azide and deoxyglucose, the amounts of CS antigen found in hepatoma cell extracts were greatly diminished. Moreover, Western blotting experiments revealed a striking difference in the pattern of CS proteins of infected cell extracts as compared with those of free parasites. The assay was used to measure the amounts of intracellular CS antigen for several days after infection of the hepatoma cells. The results confirmed previous microscopic observations, made by using immunofluorescence techniques, showing that the CS antigen in the host's liver cells diminishes progressively while the parasite develops into the exoerythrocytic stage. The immunoradiometric assay should facilitate the evaluation of the effects of drugs on sporozoites and also on studies aimed at the identification of a sporozoite receptor on the hepatocyte. PMID- 2981262 TI - Dissociation between aggregation and superoxide production in human granulocytes. AB - Aggregation and the activation of the granulocyte (PMN) superoxide (O2-) generating system occur when certain stimuli are added to resting cells. It had previously been postulated that PMN aggregation is essential for maximal O2- production. This study was undertaken to test the hypothesis that PMN aggregation is required for full expression of PMN O2- production. We examined aggregation and O2- production induced by four stimuli; concanavalin A (Con A), phorbol myristate acetate (PMA), N-formylmethionyl-leucyl-phenylalanine (FMLP), and ionophore A23187. Cytochalasin B enhanced aggregation by all four stimuli but only enhanced the rate of O2- production by Con A; 2-deoxyglucose inhibited aggregation by all stimuli. Dissociation of PMN aggregation and O2- production was achieved by using NEM, TPCK, and divalent cations. NEM and TPCK prevent Con A induced O2- production but have no effect on Con A-induced aggregation. PMA stimulated PMN generate O2- in the presence or absence of Ca++ and Mg++. In contrast, PMA stimulated maximum PMN aggregation only in the presence of both Ca++ and Mg++. Thus PMN can generate O2- without aggregating, and PMN can aggregate without producing O2-. PMN from patients with chronic granulomatous disease do not generate O2- or undergo membrane potential depolarization in response to PMA. These PMN aggregated when stimulated with PMA, providing evidence that depolarization is not required for PMN aggregation. We conclude that aggregation and the activation of the O2- generating system, though temporally related, are not necessarily causally related. PMID- 2981264 TI - Induction of neutralizing antibody in mice against poliovirus type II with monoclonal anti-idiotypic antibody. AB - Syngeneic monoclonal anti-idiotope antibody Ab2,2-17C3SCC was raised against an idiotope on a protective monoclonal antibody with specificity for poliovirus type II. Ab2,2-17C3SCC detects a paratope-related interspecies IdX. Ab2,2-17C3SCC purified from supernatant fluids of hybridoma cells by protein A-Sepharose was injected into 4- to 6-wk-old BALB/c mice. The sera of the mice were screened for the expression of antibodies bearing the corresponding idiotope. Immunization of mice with Ab2,2-17C3SCC induced antibodies of complementary specificity. Furthermore, micro VN tests suggest that Ab2,2-17C3SCC can substitute for antigen in the induction of anti-polio neutralizing antibodies, and hence can function as a monoclonal anti-idiotypic vaccine. PMID- 2981263 TI - Hemagglutination assay of polypeptide coded by the pre-S region of hepatitis B virus DNA with monoclonal antibody: correlation of pre-S polypeptide with the receptor for polymerized human serum albumin in serums containing hepatitis B antigens. AB - The receptor for polymerized human as well as chimpanzee serum albumins has been identified on the 55-amino acid polypeptide coded by the pre-S region of hepatitis B virus DNA. Monoclonal antibodies were raised against a synthetic polypeptide of 19 amino acid residues representing a hydrophilic region of the pre-S amino acid sequence deduced from hepatitis B virus DNA. Sheep erythrocytes fixed with glutaraldehyde were coated with monoclonal antibody against the synthetic polypeptide to develop a hemagglutination assay for pre-S polypeptide. The pre-S polypeptide was detected in the serum containing hepatitis B surface antigen particles along with hepatitis B e antigen, with titers in parallel with those of the receptor for polymerized human serum albumin. PMID- 2981265 TI - Protection and recovery in influenza virus-infected mice immunosuppressed with anti-IgM. AB - BALB/c mice, immunosuppressed from birth with goat anti-mouse IgM, were able to recover from influenza virus infection in the absence of detectable serum and nasal antibody. Recovery was delayed a few days when compared with control animals. Antibody-deficient mice, that had recovered from an initial influenza virus infection, i.e., convalescent mice, were subsequently rechallenged with homologous influenza virus in order to study the importance of nasal and serum antibody in prevention of infection. Convalescent mice were susceptible to reinfection when nasal and serum antibody were not detectable. The mice were resistant to reinfection when serum and/or nasal antibody was detectable by radioimmunoassay. Normal mice that were passively immunized with high titer mouse anti-influenza virus serum were susceptible to challenge with homologous influenza virus. The serum antibody levels in these mice were higher than most of those found in the immune convalescent mice suppressed with anti-IgM, thereby suggesting that the serum antibody, found in convalescent suppressed mice, is not protective. We conclude that 1) mice can recover from influenza virus infection in the absence of detectable levels of nasal and serum antibody, thus indirectly confirming the role of cell-mediated immunity in recovery; 2) serum IgM, IgG2A, IgG2B, IgG3, and probably IgG1 antibody levels are not responsible for protection against influenza virus infection of the upper respiratory tract; and 3) nasal IgA antibody correlates best with protection against reinfection of the upper respiratory tract, but some other locally protective agent cannot be excluded. PMID- 2981266 TI - Organization and polymorphism of rabbit immunoglobulin heavy chain genes. AB - Germline genes encoding C mu, C gamma, C alpha, and C epsilon heavy chains of rabbit immunoglobulins have been isolated from recombinant phage and cosmid libraries. The JH, C mu, C gamma, and C epsilon are found in a 5'-JH-C mu-C gamma C epsilon-3' orientation on a 90kb stretch of DNA. Four C alpha genes have been cloned and presumably reside 3' to the other CH genes. Southern blot analysis of rabbit sperm DNA indicates that the rabbit genome contains a single C gamma gene, one C mu gene, and as many as 10 C alpha genes. Restriction site polymorphism is found for C mu, C gamma, and C alpha genes of rabbits of various heavy chain haplotypes. The organization of the rabbit CH genes differs from that of mouse and human CH genes in that the rabbit has multiple C alpha genes, whereas mouse and human have one or two C alpha genes, respectively. In addition, mouse and human have four C gamma genes, whereas rabbit has only one C gamma gene. The presence of a single C gamma gene indicates that at least in the rabbits examined, no germline gene encoding latent or unexpected, C gamma allotypes is present. The genetic control of the expression of latent C gamma allotypes is discussed. PMID- 2981267 TI - Analysis of the rat major histocompatibility system by Southern blot hybridization. AB - The organization of the rat major histocompatibility complex, RT1, was studied at the DNA level by Southern blot hybridization. Genomic DNA from eight different RT1 congenic rat strains was digested by various restriction enzymes and was hybridized under stringent conditions with probes of mouse class I and class II H 2 genes. Few cross-hybridizing DNA fragments, showing no polymorphism, were seen with class II A alpha and A beta probes. The class I probes allowed for the distinction of about 8 to 19 cross-hybridizing bands, which exhibited extensive polymorphism. With the use of five RT1 recombinants, about 20% of the DNA fragments could be mapped to the RT1.A region, which codes for the ubiquitously expressed class I antigens, and about 80% to the RT1.C region-determining class I like antigens, which are different from RT1.A antigens with respect to tissue distribution, restriction function in immune responses, and allograft rejection. The number of class I genes present in the rat genome and the possible relationship of RT1.C to H-2Qa, Tla of the mouse are discussed. PMID- 2981268 TI - Specific effect of anti-transferrin antibodies on natural killer cells directed against tumor cells. Evidence for the transferrin receptor being one of the target structures recognized by NK cells. AB - Treatment of PBL or Percoll-isolated LGL with anti-transferrin antibodies plus complement reduced their natural killing activity against K-562 cells between 30 and 70%. The same antibodies inhibited natural cytotoxicity when added directly to the assay. Similar depletion or inhibition of NK cytotoxicity was observed when using HeLa cells as targets. The decrease or inhibition by transferrin antibodies was less marked when IFN-treated PBL or LGL as effector cells were used. The inhibition of anti-transferrin antibodies seems to be located at the level of the effector cell population. When PBL but not target K-562 cells were pretreated with anti-transferrin antibodies and were washed before use in the assay, cytotoxicity was decreased by 50%. In addition, about 80% of the LGL positively selected on anti-transferrin plates stained with Leu-11. Furthermore, no reduction by anti-transferrin antibodies plus complement treatment of PBL or LGL, or inhibition by antibodies alone, was observed when the cells were tested against HSV-1-infected cells. Membrane extracts from LGL inhibited NK cytotoxicity against K-562 or HeLa cells. Moreover, the inhibitory component of this extract was removed by anti-transferrin IgG but not by control IgG. These results are in agreement with the recent hypothesis that NK cells recognize the transferrin receptor in tumor target cells, because both the transferrin receptor and anti-transferrin antibodies may share a similar structure that interacts with the NK cells. PMID- 2981269 TI - Tac antigen-positive T cells activated in autologous mixed lymphocyte reaction regulate the generation of killer T cells against hapten-modified autologous cells. AB - T cells that proliferate in the autologous mixed lymphocyte reaction (auto-MLR) have been shown to acquire some suppressor or regulatory activities. In the present study, we examined the suppressive effects of T cells activated in the auto-MLR on the induction of hapten-specific cytotoxic T cells. NRFT (depletion of ARFT from UT) were used as the responder cells of TNP-MLR. After primary and secondary TNP-MLR, the cells were harvested and tested for their cytotoxic activities against TNP-modified autologous cells by 51Cr-release assay. When UT cells cultured for 1 wk in auto-MLR were added to primary TNP-MLR at the beginning of culture, the cytotoxic activity tested at the end of the culture was suppressed from 15.6% +/- 2.7 to 5.8% +/- 1.1 (percent cytotoxicity, mean +/- SE). However, these auto-MLR-activated UT cells had little suppressive activity against cytotoxic T cells when they were added to the final assay of TNP-CTR. Suppressive activities of these cells on the generation of cytotoxic T cells during secondary TNP-MLR were also tested. The addition of auto-MLR-activated UT cells to the secondary TNP-MLR at the beginning of the culture reduced the cytotoxic activities of NRFT from 23.8% +/- 2.3 to 9.7% +/- 1.7 after secondary TNP-MLR. Allo-activated T cells, PHA blasts, and fresh autologous T cells were used as the controls, but none of the cells had suppressive effects on the generation of CTL. Characteristics of these suppressor cells were examined. Auto MLR-activated cells from ARFT fractions exhibit very powerful suppressor activity. Treatment of the auto-MLR-activated T cells with mitomycin C eliminated their suppressive effects on the generation of CTL; 21.2% +/- 6.3 of UT cells became anti-Tac positive after 1 wk of auto-MLR. Treatment of auto-MLR-activated UT cells with anti-Tac antibody plus complement eliminated their suppressive activities on the induction of CTL. Thus, T cells stimulated in auto-MLR were shown to have suppressive effects on the induction of cytotoxic T cells against TNP-modified autologous cells. These cells were mitomycin C sensitive. Because anti-Tac antibody is reactive to activated T cells, activation of T cells during auto-MLR was thought to be necessary for the acquisition of the suppressive activity. PMID- 2981270 TI - Augmentation of immunity to herpes simplex virus by in vivo administration of interleukin 2. AB - The immune mechanisms responsible for recovery from herpesvirus infections are multiple and include a principle role for aspects of T cell immunity. Our investigations add further support for this notion. We show that the ability of immune lymphocytes from animals infected i.p. 6 wk previously with herpes simplex virus type one (HSV-1) clear virus more effectively when interleukin 2 (IL 2) is injected into recipients of the adoptive transfers. Mice were treated on two consecutive days with cyclophosphamide and infected in the pinnae with 4 X 10(6) plaque-forming units of HSV-1. Three hours post-infection lymphocyte populations were injected i.v., and after a further 3 days the pinnae were removed, homogenized, and the content of infectious virus assayed. Purified IL 2 obtained from EL-4 cells either was given i.v. 2 hr before and 24 and 48 hr after cell injection or was given subcutaneously 2 hr before and 3, 24, and 48 hr after cell injection. The latter three injections were given i.p. and suspended in 15% gelatin. The immune lymphocyte cell populations were splenocytes and were either injected immediately after preparation of cell suspensions or after 5 days in vitro secondary stimulation with HSV-1. This latter cell population showed greater viral clearance activity, a function shown previously to be a property of Lyt-2+ cells. The clearance activity of cells was markedly enhanced in animals given IL 2 but only with a regimen that included injections in gelatin, a procedure that enhances in vivo circulation time of IL 2. The cell involved in clearance was a T cell and principally the Lyt-2+ subset. Treatment of recipient mice with anti-asialo GM-1 did not affect the clearance efficiency, indicating that NK cells were not responsible for the observed effect. Our experiments indicate that IL 2 may provide an important regulator of immune function in vivo and may warrant its investigation as a therapeutic agent to enhance antiviral immunity in certain circumstances. PMID- 2981271 TI - Regulation by interleukin 2 of interleukin 2 receptors and gamma-interferon synthesis by human thymocytes: augmentation of interleukin 2 receptors by interleukin 2. AB - The role of interleukin 2 (IL 2) on the expression of IL 2 receptors and on the synthesis of gamma-interferon (gamma-IFN) by human thymocytes was investigated. Human thymocytes isolated from specimens obtained during cardiac surgery of infants and children were induced with one or all of the following agents: IL 2, concanavalin A (Con A), and 12-O-tetradecanoylphorbol 13-acetate (TPA). The expression of IL 2 receptors and gamma-IFN titers were determined. The results indicate that thymocytes cultured in complete medium do not express receptors for IL 2, nor did IL 2 by itself induce the expression of IL 2 receptors. Con A induced the expression of IL 2 receptors by a moderate number of the thymocyte population and induced the synthesis of low amounts of gamma-IFN. Preincubation of thymocytes with TPA increased the response to Con A; both the number of thymocytes expressing receptors and the synthesis of gamma-IFN were increased. Addition of IL 2 to these cultures further augmented the expression of IL 2 receptors and gamma-IFN synthesis and resulted in the optimal expression of IL 2 receptors and maximal gamma-IFN synthesis. The expression of IL 2 receptors could be detected within 24 hr and preceded the induction of proliferation; it was therefore probably not due to the clonal expansion of a population of receptor bearing thymocytes. Conversely, inhibition of IL 2 synthesis with dexamethasone (Dex) by thymocytes activated with Con A, or inhibition of the function of IL 2 receptors by anti-Tac, resulted in a decrease in the number of IL 2 receptor bearing thymocytes activated with Con A, or inhibition of the function of IL 2 receptors by anti-Tac, resulted in a decrease in the number of IL 2 receptor bearing thymocytes and of gamma-IFN synthesis. Thymocytes activated with TPA and Con A were more resistant to the inhibitory effects of Dex on the expression of IL 2 receptors than thymocytes activated with Con A alone. Maximal inhibition of the expression of IL 2 receptors and of gamma-IFN synthesis was achieved as a result of the synergistic effect of anti-Tac with Dex. Therefore, when IL 2 was prevented from binding to the receptors, and IL 2 synthesis was inhibited, the number of thymocytes expressing IL 2 receptors was sharply reduced and gamma-IFN synthesis was markedly inhibited.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981272 TI - Effect of recombinant IL 2 and gamma-IFN on proliferation and differentiation of human B cells. AB - The effects of IL 2 and gamma-IFN on the activation of human B cells was studied with recombinant IL 2 and gamma-IFN. BCDF-responsive B lymphoblastoid cell lines and highly purified human B cells were employed as target B cells. IL 2 or gamma IFN did not induce any IgG or IgM secretion in the B cell lines CESS and SKW6 CL4, in which IgG and IgM were inducible with conventional T cell factor(s). IL 2 alone did not induce the optimum production of Ig, but did induce proliferation in the SAC-stimulated B cell population. No Leu-1-, Leu-4-, or Leu-7-positive cells were detected in B cell populations that had been stimulated with SAC for 3 days. FACS analysis showed that a portion of the SAC-stimulated B cells (30%) were in the G2 or M stages by IL 2 stimulation. The addition of gamma-IFN together with IL 2 induced IgM and IgG secretion in SAC-stimulated B cells that was comparable with that induced by a conventional T cell factor(s). IL 2 induced proliferation not only in SAC-stimulated B cells but also in an anti-mu stimulated B cell population. Stimulation of T cell populations with anti-mu and IL 2 did not induce significant proliferation, suggesting the direct effect of IL 2 on B cells. Double staining of anti-mu-stimulated B cells with anti-Ig and anti Tac antibodies demonstrated that anti-mu stimulation induced an increased expression of Tac antigen on surface Ig-positive B cells. All of these results strongly supported the notion that IL 2 was one of the growth factors for B cells, and gamma-IFN was one of the differentiation factors for B cells. PMID- 2981273 TI - Collagen biosynthesis in systemic scleroderma: regulation of posttranslational modifications and synthesis of procollagen in cultured fibroblasts. AB - Activities of prolyl hydroxylase (PH), lysyl hydroxylase (LH), and the collagen glycosyltransferases and the extent of the posttranslational modification of lysine residues in newly synthesized collagen were studied in fibroblast cultures obtained from 9 scleroderma patients. The rate of procollagen synthesis had increased more than 3-fold in 3 scleroderma fibroblast lines, but had not changed to the same extent in the others, even though these did not differ from the "high producers" histologically, clinically, or immunohistologically. The activities of PH and LH correlated significantly with the rate of procollagen synthesis in the same cell lines (p less than 0.001), but the glycosyltransferase activities were not elevated in the scleroderma fibroblasts. Further studies nevertheless indicated that the extent of the posttranslational modification of lysine residues had not significantly changed in the procollagen synthesized by any of the scleroderma fibroblasts investigated. PMID- 2981274 TI - Varicella-zoster virus infection of strain 2 guinea pigs. AB - Weanling strain 2 guinea pigs are susceptible to infection with varicella-zoster virus cultured in embryonic guinea pig tissue. Animals inoculated by an intramuscular route develop mononuclear cell viremia that may persist for as long as three weeks. During the period of viremia, virus may be recovered from the nasopharynx and a variety of tissues. In addition, virus may be recovered from neural tissues in the absence of viremia, although infectious virus has not been cultivated from neural tissues after the 23rd day. The strain 2 guinea pig should provide an animal model to study the pathophysiology of infections caused by varicella-zoster virus. PMID- 2981275 TI - Influence of drugs that block calcium channels on the microbicidal function of human neutrophils. AB - The central role of calcium ions in cell physiology prompted us to examine the hypothesis that pharmacological concentrations of calcium channel-blocking drugs might affect human neutrophil (PMN) functions. The capacity of PMNs suspended in verapamil hydrochloride for killing Pseudomonas aeruginosa during two hour incubations was significantly impaired (P less than .05). Several observations suggested that this drug effect was the result of altered calcium metabolism: exposure to verapamil decreased the uptake of 45Ca++ by PMNs subsequently exposed to the calcium ionophore A23187; verapamil did not impair PMN function in the absence of extracellular calcium; and the addition of A23187 concomitantly with (but not following) verapamil prevented PMN dysfunction. In addition, nifedipine, a structurally dissimilar calcium channel-blocking drug, also impaired the bactericidal activity of PMNs against Pseudomonas aeruginosa (P less than .02). Further studies revealed that treatment with verapamil did not affect PMN phagocytosis, but significantly impaired the PMN respiratory burst (as shown by superoxide anion generation assay; P less than .05). We conclude that PMNs exposed to pharmacological concentrations of calcium channel-blocking drugs exhibit a reduced capacity to kill bacteria. PMID- 2981277 TI - Defective triggering of polymorphonuclear leukocyte oxidative metabolism by Legionella pneumophila toxin. AB - Preincubation of human polymorphonuclear leukocytes (PMNLs) with Legionella pneumophila toxin impaired activation of the superoxide-generating complex induced by latex particles and by the Ca++ ionophore A23187. The toxin had no effect, however, on activation of the complex induced by phorbol myristate acetate (PMA), concanavalin A, valinomycin, or bromolasalocid. The toxin prevented PMNL plasma membrane depolarization induced by A23187 but failed to influence the membrane depolarization induced by PMA. These observations indicate that Legionella pneumophila toxin selectively impairs activation of the phagocyte superoxide-generating complex without affecting the functional integrity of components of the complex. PMID- 2981278 TI - Evidence of immunity induced by naturally acquired rotavirus and Norwalk virus infection on two remote Panamanian islands. AB - For better understanding of the role of humoral immunity in ameliorating infections with rotavirus (RV) and Norwalk virus (NW), 305 Cuna Indians living on two isolated islands located off Panama's Carribean coast were surveyed daily for diarrhea over a seven-month period. Nine (8%) of 108 persons with a baseline RV antibody titer of greater than 1:4 developed RV infection compared with 70 (46%) of 151 persons with a baseline RV antibody titer of less than 1:4 (P less than .001). Thirty-eight (25%) of 151 persons of all ages with baseline RV antibody titer of less than 1:4 had at least one episode of RV diarrhea compared with 6 (6%) of 108 persons who had baseline RV antibody titers of greater than 1:4 (P less than .001). Thirty-two (47%) of 68 persons of all ages who had a baseline NW antibody titer of less than 1:100 developed NW infection compared with 30 (13%) of 237 persons with a baseline NW titer of greater than 1:100 (P less than .001). The high NW and RV infection rates and the excellent levels of protection provided by specific preexisting humoral antibody to these agents should promote activities aimed at developing vaccines for preventing these infections. PMID- 2981276 TI - Response of children of patients with primary hepatocellular carcinoma to hepatitis B virus vaccine. PMID- 2981279 TI - The modification of the oxidative metabolism of cells derived both locally and at distance from the site of an acute inflammatory reaction. AB - During an acute nonspecific inflammatory reaction initiated in the pleural cavity by a nondiffusible stimulus (calcium pyrophosphate crystals), the oxidative metabolism, as measured by chemiluminescence and superoxide release, of cells harvested from both the inflammatory site and at points distant from it was studied. The oxidative metabolism of peritoneal macrophages, obtained from rats undergoing an inflammatory reaction (pleurisy), demonstrated a transient decrease in activity compared with the resident population when using both zymosan and phorbol myristate acetate as stimulants. This metabolic unresponsiveness induced by inflammation may be related to the concomitant changes in the levels of prostacyclin in the peritoneal cavity. It should be emphasized that the peritoneal cellular composition or number did not change during these events. On the other hand alveolar macrophages from inflamed animals showed no significant changes in their superoxide production or chemiluminescence compared to controls. The precise reason for these inflammation-induced changes is unknown; however the acute nonspecific inflammatory reaction was able to modulate the oxidative metabolism of cells not only at the site of inflammation, but at points distant from it. PMID- 2981280 TI - Inhibition of the electrically evoked release of [3H]acetylcholine in rat striatal slices: an experimental model for drugs that enhance dopaminergic neurotransmission. AB - The activation by endogenous dopamine of the inhibitory 3,4 dihydroxyphenylethylamine (dopamine) receptors modulating the electrically evoked release of [3H]acetylcholine [( 3H]ACh) and [3H]dopamine in rat striatal slices is a function of the concentration of dopamine accumulated in the synaptic cleft during electrical stimulation. When the release of 3H-neurotransmitters was elicited with a 2-min period of stimulation at a frequency of 1 Hz, neither dopamine autoreceptors nor dopamine receptors modulating [3H]ACh were activated by endogenously released dopamine. On the other hand, exposure to (S)-sulpiride facilitated the release of [3H]dopamine and [3H]ACh elicited when the 2-min stimulation was carried out at a frequency of 3 Hz but this effect was not observed at a lower frequency of stimulation (1 Hz). In the presence of amphetamine the dopamine receptors modulating the electrically evoked release of [3H]ACh can be activated by endogenous dopamine even at the lower frequency of stimulation (1 Hz). Similar effects can be obtained if the neuronal uptake of dopamine is inhibited by cocaine or nomifensine. The inhibition by amphetamine of the release of [3H]ACh elicited by electrical stimulation at 1 Hz involves dopamine receptors and can be fully antagonized by clozapine, haloperidol, chlorpromazine, or pimozide. The stereoselectivity of this antagonism can be demonstrated with the optical enantiomers of sulpiride and butaclamol. This inhibitory effect of amphetamine on cholinergic neurotransmission appears to be the result of the stimulation of dopamine receptors of the D2 subtype, as they were resistant to blockade by the preferential D1 receptor antagonist SCH 23390.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981281 TI - Decrease of clonidine binding affinity to alpha 2-adrenoceptor by ADP ribosylation of 41,000-dalton proteins in rat cerebral cortical membranes by islet-activating protein. AB - The IC50 value for inhibition of specific [3H]yohimbine binding to rat cerebral cortical membranes by clonidine was increased, and the Hill coefficient (nH) approached unity in the presence of 150 microM GTP. Pretreatment of membranes with islet-activating protein (IAP) in the presence of NAD caused an increase in IC50 and nH values for clonidine compared with control membranes in the absence of GTP, the addition of which was without effect. Scatchard analysis showed that the Bmax value of the high-affinity component in [3H]clonidine binding was decreased by pretreatment with IAP/NAD. GTP in a concentration range of 0.1 microM-1 mM caused a significant elevation of [3H]yohimbine binding. In IAP/NAD pretreated membranes, however, [3H]yohimbine binding was no longer affected by GTP, although IAP/NAD significantly (p less than 0.01) increased [3H]yohimbine binding compared to control. IAP ADP-ribosylated 41,000 dalton proteins of cerebral cortical membranes. From these results, it can be suggested that inhibitory guanine nucleotide regulatory protein with Mr 41,000 couples to alpha 2-adrenoceptors to regulate binding affinity of agonists and antagonists in membranes of the rat cerebral cortex. PMID- 2981282 TI - Muscarinic receptors and hydrolysis of inositol phospholipids in rat cerebral cortex and parotid gland. AB - Exposure of rat brain or parotid gland slices to muscarinic receptor agonists stimulates a phospholipase C that degrades inositol phospholipids. When tissue slices were labelled in vitro with [3H]inositol, this response could be monitored by measuring the formation of [3H]inositol phosphates. Accumulation of inositol 1,4-biphosphate in stimulated brain slices suggests that polyphosphonositides are the primary targets for phospholipase C activity. Li+ (10 mM) in the medium completely blocked the hydrolysis of inositol 1-phosphate, partially inhibited inositol 1,4-bisphosphate hydrolysis, but had no effect on the hydrolysis of inositol 1,4,5-trisphosphate by endogenous phosphatases. Muscarinic receptor pharmacology was studied by measuring the accumulation of [3H]inositol 1 phosphate in the presence of 10 mM Li+. In experiments on brain slices, the response to carbachol was antagonised by atropine with an affinity constant of approximately 8.79 +/- 0.12. Dose-response curves to several muscarinic agonists were constructed using brain and parotid gland slices. The results are consistent with relatively direct coupling of low-affinity muscarinic receptors to inositol phospholipid breakdown in brain slices; full agonists were relatively more potent in the parotid gland compared with the brain. Explanations for these differences are suggested. PMID- 2981283 TI - Dissociation of [35S]t-butylbicyclophosphorothionate binding differentiates convulsant and depressant drugs that modulate GABAergic transmission. AB - The dissociation of [35S]t-butylbicyclophosphorothionate ([35S]TBPT) from binding sites on membranes from rat cerebral cortex, after addition of saturating concentrations of convulsant and depressant drugs, was studied. The addition of unlabeled TBPT, picrotoxinin, or pentamethylenetetrazol resulted in dissociation patterns that were monophasic and not distinguishable, suggesting that these convulsants bind competitively to the same (convulsant) sites. In contrast, gamma aminobutyric acid (GABA) greatly facilitated [35S]TBPT dissociation by binding allosterically to the GABA recognition site of the receptor-ionophore complex. TBPT dissociation was similarly accelerated by the depressants etazolate, (+) etomidate, and barbiturates. The convulsant and depressant S(+) and R(-) stereoisomers of N-methyl-5-phenyl-5-propyl-barbituric acid displayed large stereoselectivity in the acceleration of TBPT dissociation. These results suggest that depressants bind to sites different from the convulsant sites of the allosteric GABA receptor complex, or the binding of depressants to the same population of sites elicits negative cooperativity and dissociates the convulsants. PMID- 2981284 TI - Regulation of dopamine release from PC12 pheochromocytoma cell cultures during stimulation with elevated potassium or carbachol. AB - To examine the role of cyclic AMP in the process of catecholamine release experiments have been performed with cultures of PC12 pheochromocytoma cells. Elevated potassium (56 mM) and carbamylcholine (carbachol, 10(-4) M) cause rapid increases in cyclic AMP levels in the cultures that show a time course similar to that of evoked dopamine release. These secretogogue-induced increases in cyclic AMP levels are well correlated with release in terms of relative magnitude and calcium dependence. Forskolin (a direct activator of adenylate cyclase) causes dose-related increases in cyclic AMP levels in PC12 cell cultures that are synergistic with those caused by either elevated potassium or carbachol. At low concentrations forskolin significantly increases evoked release, whereas at higher concentrations it increases both spontaneous and evoked release. These results suggest that cyclic AMP may be involved in the process of dopamine release from PC12 cells in culture. PMID- 2981285 TI - An immunoblot study of neurofilament degradation in situ and during calcium activated proteolysis. AB - The degradation of neurofilament (NF) proteins was examined by immunoblot methods to identify, characterize, and monitor the appearance of immunoreactive breakdown products during the loss of NF triplet proteins. Individual NF proteins and their breakdown products were identified using polyclonal and monoclonal antibodies to NF proteins. NF degradation was compared during calcium-activated proteolysis of isolated rat NF, during an experimental influx of calcium into excised rat spinal nerve roots, and during NF breakdown in transected rat peripheral nerve. These different experimental conditions produced similar patterns of NF fragmentation, including the transient appearance of NF immunobands between Mr 150,000-200,000 and 110,000-120,000 as well as the appearance and accumulation of NF immunobands between Mr 45,000 and 65,000. Most immunoreactive NF fragments remained Triton insoluble. Low levels of the same immunoreactive fragments were present in control neural tissues, suggesting that calcium-activated proteolysis may be operative in the turnover and/or processing of NF proteins in vivo. Very similar patterns of NF degradation during experimental calcium influxes into different CNS and PNS tissues are indicative of the widespread distribution of calcium activated NF protease in neural tissues. PMID- 2981286 TI - Inhibition of a low Km GTPase activity in rat striatum by calmodulin. AB - In rat striatum, the activation of adenylate cyclase by the endogenous Ca2+ binding protein, calmodulin, is additive with that of GTP but is not additive with that of the nonhydrolyzable GTP analog, guanosine-5'-(beta, gamma imido)triphosphate (GppNHp). One possible mechanism for this difference could be an effect of calmodulin on GTPase activity which has been demonstrated to "turn off" adenylate cyclase activity. We examined the effects of Ca2+ and calmodulin on GTPase activity in EGTA-washed rat striatal particulate fractions depleted of Ca2+ and calmodulin. Calmodulin inhibited GTP hydrolysis at concentrations of 10( 9)-10(-6) M but had no effect on the hydrolysis of 10(-5) and 10(-6) M GTP, suggesting that calmodulin inhibited a low Km GTPase activity. The inhibition of GTPase activity by calmodulin was Ca2+-dependent and was maximal at 0.12 microM free Ca2+. Maximal inhibition by calmodulin was 40% in the presence of 10(-7) M GTP. The IC50 for calmodulin was 100 nM. In five tissues tested, calmodulin inhibited GTP hydrolysis only in those tissues where it could also activate adenylate cyclase. Calmodulin could affect the activation of adenylate cyclase by GTP in the presence of 3,4-dihydroxyphenylethylamine (DA, dopamine). Calmodulin decreased by nearly 10-fold the concentration of GTP required to provide maximal stimulation of adenylate cyclase activity by DA in the striatal membranes. The characteristics of the effect of calmodulin on GTPase activity with respect to Ca2+ and calmodulin dependence and tissue specificity parallel those of the activation of adenylate cyclase by calmodulin, suggesting that the two activities are closely related.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981287 TI - Calcium-dependent calmodulin binding to chromaffin granule membranes: presence of a 65-kilodalton calmodulin-binding protein. AB - The presence of calmodulin-binding sites on chromaffin granule membranes has been investigated. Saturable, high-affinity 125I-calmodulin-binding sites (KD = 9.8 nM; Bmax = 25 pmol/mg protein) were observed in the presence of 10(-4) M free calcium. A second, nonsaturable, calmodulin-binding activity could also be detected at 10(-7) M free calcium. No binding occurred at lower calcium levels. When chromaffin granule membranes were delipidated by solvent extraction, calmodulin binding was observed at 10(-4) M free calcium. However no binding was detected at lower calcium concentrations. Thus it appears that a calcium concentration of 10(-7) M promotes the binding of calmodulin to some solvent soluble components of the chromaffin granule membrane. Calmodulin-binding proteins associated with the granule membrane identified by photoaffinity cross linking. A calmodulin-binding protein complex, of molecular weight 82K, was formed in the presence of 10(-4) M free calcium. This cross-linked product was specific because it was not detected either in the absence of calcium, in the presence of nonlabeled calmodulin, or in the absence of cross-linker activation. When solvent-treated membranes were used, a second, specific, calmodulin-binding protein complex (70K) was formed. Since the apparent molecular weight of calmodulin in our electrophoresis system was 17K, these experiments suggested the presence of two calmodulin-binding proteins, of molecular weights 65K and 53K, in the chromaffin granule membrane. This result was confirmed by the use of calmodulin-affinity chromatography. When detergent-solubilized membranes were applied on the column in the presence of calcium, two polypeptides of apparent molecular weights of 65K and 53K were specifically eluted by EGTA buffers. Since detergent treatments or solvent extractions are necessary to detect the 53K calmodulin-binding protein, it is concluded that only the 65K calmodulin-binding polypeptide may play a role in the interaction between calmodulin and secretory granules in chromaffin cells. PMID- 2981288 TI - Tetraenoic species are conserved in muscarinically enhanced inositide turnover. AB - Carbamylcholine enhances the labeling of phosphatidate and phosphatidylinositol from 32Pi in nerve endings. Approximately 74% of labeled phosphatidate and 85% of labeled phosphatidylinositol produced on muscarinic stimulation are accounted for by tetraenoic species, as detected by argentation TLC. Incubation of membranes derived from nerve endings with [gamma-32P]ATP under conditions of phosphodiesteratic degradation of endogenous polyphosphoinositides resulted in increased labeling of phosphatidate. Approximately 78% of the newly formed phosphatidate was in a tetraenoic fraction. It is concluded that in muscarinically stimulated nerve endings, the diacylglycerol moiety is conserved following diacylglycerol release from polyphosphoinositides through its resynthesis to inositol lipid via phosphatidate. PMID- 2981289 TI - Regulation of phosphate incorporation into four brain phosphoproteins that are affected by experience. AB - Various regulators of protein kinase activities were tested for their effects on the in vitro transfer of phosphate from [gamma-32P]ATP to four proteins of rat brain synaptic particulate preparations. One protein, of apparent molecular weight 44,000, accepted 32P in the presence of 8 mM EDTA and no added Mg2+. It was the major phosphoprotein of brain mitochondria. Its phosphorylation was inhibited by pyruvate and stimulated by K+, and it comigrated in electrophoretic gels with authentic alpha-subunit of pyruvate: lipoamide oxidoreductase (decarboxylating) (EC 1.2.4.1) from bovine heart. The major kinase acting on three proteins of apparent molecular weights 24,000, 21,000, and 19,000 was stimulated by Ca2+, by preincubation with phospholipase C, and by 12 tetradecanoyl 4-beta-phorbol 13-acetate. Phosphorylation of these lower-molecular weight proteins was inhibited by ACTH1-24, by cyclic 3',5'-adenosine monophosphate, and by 50 microM trifluoperazine. The stimulatory effect of Ca2+ was antagonized by calmodulin. The kinase in question appears to be B-50 protein kinase or protein kinase C. PMID- 2981290 TI - Correlation between 2',3'-cyclic nucleotide 3'-phosphohydrolase activity and demyelination in vitro using a syngeneic system. AB - Cultures of myelinated SJL/J fetal mouse spinal cord were incubated with serum and lymphoid cells from syngeneic animals with experimental allergic encephalomyelitis (EAE) induced by syngeneic spinal cord homogenate (SSCH) in complete Freund's adjuvant or others injected with complete Freund's adjuvant alone. After 24 or 48 h of exposure, demyelination was determined by light microscopic examination and quantification of 2',3'-cyclic nucleotide 3' phosphohydrolase activity. Cultures exposed to spleen or lymph node cells from SSCH-sensitized animals showed the greatest alterations in myelin and decreases in 2',3'-cyclic nucleotide 3'-phosphohydrolase activity whereas serum from these animals had less effect. Cells and serum from complete Freund's adjuvant-injected control animals also induced structural changes in myelin that were significantly less than changes induced by cells and serum from animals with EAE. These experiments show that lymphoid cells and serum obtained from SJL/J mice with acute EAE affected myelin biochemistry and morphology in syngeneic CNS cultures. PMID- 2981291 TI - Cerebrospinal fluid vasopressin as a marker of central nervous system metastases from small-cell bronchogenic carcinoma. AB - Vasopressin (ADH) was measured in CSF and plasma in 75 evaluable patients with known or suspected CNS metastases from small-cell bronchogenic carcinoma (SCBC), and in 66 control patients having neither malignant disease nor organic CNS disease. The presence of CNS metastases was confirmed or excluded on the basis of computed tomographic scans, neurologic examination, and autopsy. Twenty-four of the 75 patients had no CNS metastases. Ten of the 51 patients with CNS metastases had leptomeningeal carcinomatosis (MC). CSF-ADH was significantly increased in patients with MC (P less than .05), but not in patients having exclusively parenchymatous CNS metastases. Taking 2 pg/mL (95th percentile of control patients) as the upper limit of normal, 15 SCBC patients had elevated CSF-ADH, including 12 patients with CNS metastases and six patients with MC. The CSF-ADH to plasma ADH ratio was significantly increased in patients with CNS metastases (P less than .05). Patients without CNS metastases had a ratio less than or equal to 0.8 whereas the ratio was greater than 0.8, in 21 of the 51 patients with CNS metastases. The positive and negative predictive values with 95% confidence limits were 84% to 100% and 31% to 59%, respectively. Patients with inappropriate secretion of ADH (SIADH) constituted a significantly greater proportion of patients with elevated CSF-ADH than of patients with normal CSF-ADH levels (P less than .05). In addition, patients with SIADH constituted a significantly greater proportion of patients with MC than of patients with parenchymatous metastases (P less than .05). The diagnostic application of these findings is limited because of the large number of false-negative results, but it may prove to be of value in conjunction with the measurement of other tumor markers. PMID- 2981293 TI - Etoposide (VP-16) and cisplatin: an effective treatment for relapse in small-cell lung cancer. AB - Seventy-eight patients with evaluable small-cell lung cancer (SCLC) were treated with etoposide (VP-16) and cisplatin after their disease failed to respond to, or relapsed after, induction combination chemotherapy, consisting primarily of cyclophosphamide, doxorubicin (Adriamycin), and vincristine (CAV). Twenty-four patients had limited disease (LD) and 54 had extensive disease (ED). In six (8%) patients, a complete response (CR) was achieved and in 37 (47%), there was a partial response (PR). The median duration of response for responding patients was 22 weeks (range, 4 to 50 weeks) for patients with LD and 18 weeks (range, 4 to 49 weeks) for those with ED. Twelve percent of patients demonstrated stable disease, and 33% of patients had progressive disease on treatment. The median survival times of LD patients achieving a CR or PR were 59 and 34 weeks, respectively, whereas the comparable figures for ED patients were 45 and 23 weeks, respectively. Gastrointestinal toxicity was mild, but myelosuppression, predominantly leukopenia and thrombocytopenia, was common. Mild to moderate nephrotoxicity occurred in 11 patients, but was reversible in all cases. Two febrile episodes occurred during periods of drug-induced neutropenia, but no other significant toxicities were identified. These results provide further evidence that VP-16 and cisplatin is an effective and tolerable combination chemotherapy regimen for SCLC resistant to CAV. PMID- 2981292 TI - Co-trimoxazole prophylaxis during high-dose chemotherapy of small-cell lung cancer. AB - In 103 patients with small-cell lung cancer, we compared four courses of standard doses of Adriamycin (A) (Adria Laboratories, Columbus, Ohio), vincristine (V), and cyclophosphamide (C) with a regimen of increased doses of cyclophosphamide and to a lesser extent, Adriamycin. We found no significant difference in rate (22% v 21%) or median duration (seven v nine months) of complete remission. Patients not in complete remission after the four cycles of AVC received two courses of VP-16 (etoposide) and cisplatin: the complete remission rate increased to 49% and 48% respectively. Patients on the high-dose arm received co trimoxazole prophylaxis; those on the standard arm did not. Patients on the high dose arm had a higher incidence of neutropenia (nadir less than 500 cells/microL) but a lower incidence of infection for similar degrees of neutropenia. However, they also suffered more severe side effects of a different kind. Cotrimoxazole thus allowed for the administration of higher doses of chemotherapy to outpatients by protecting them from infection. However, the higher doses of cyclophosphamide and Adriamycin, did not improve treatment results, produced more severe side effects, and is not recommended. PMID- 2981294 TI - Long-term survival in small-cell carcinoma of the lung: a population experience. AB - Small-cell lung carcinoma (SCLC) is a rapidly progressive and fatal disease. Historically, surgical resection or radiotherapy of the primary tumor has done little to prolong survival, although the use of combination chemotherapy is more effective. Reported here is the survival experience of 1,538 incident cases of SCLC identified through the Surveillance, Epidemiology and End Results Program in western Washington State from 1974 to 1982. The survival experience of this population series is similar to that reported from specialized referral centers. For 71 of 78 persons surviving at least 24 months, the original diagnostic slides were independently reviewed, 47 cases being confirmed as SCLC. No differences were found in actuarial survival estimates between those confirmed and those not confirmed as SCLC. Multivariate survival analysis was conducted to estimate the effects on survival of stage, therapy, age, sex, primary site, and histologic type. All factors except primary site and histologic type significantly influence initial survival rates. However, the only factor related to post--two-year (ie, long-term) survival, once stage is accounted for, is whether surgery was received as a first course of therapy. Those not receiving surgery were at four times the risk of death as those who did. These results indicate that long-term survival can be achieved in patients with SCLC treated in the community, and that the chance of surviving an additional two years for such patients is approximately 40%. PMID- 2981295 TI - Postsynaptic block of frog neuromuscular transmission by conotoxin GI. AB - Conotoxin GI, a peptide neurotoxin contained in the venom of the marine snail Conus geographus, was applied to the cutaneous pectoris muscle of the frog, and the effects on the postsynaptic response to acetylcholine were examined. Conotoxin GI reversibly blocked nerve-evoked muscle contractions at concentrations greater than or equal to 3 to 4 microM. Micromolar concentrations of conotoxin GI significantly reduced the amplitude of miniature endplate potentials and membrane depolarizations produced by ionophoretic application of acetylcholine, suggesting that the toxin reduced the postsynaptic sensitivity to acetylcholine. The reduction in the sensitivity of the muscle to acetylcholine was not due to changes in muscle fiber resting membrane potential or input resistance. Conotoxin GI reduced the amplitudes but did not affect the rates of decay of focal, extracellularly recorded endplate currents or miniature endplate currents, suggesting that the toxin did not affect the lifetime of ion channels opened by acetylcholine. Miniature endplate currents decay five to six times more slowly than normal when acetylcholinesterase is blocked with neostigmine methyl sulfate due to repeated binding of acetylcholine to receptors as it diffuses from the synaptic cleft. Conotoxin GI reduced the amplitude and increased the rate of decay of miniature endplate currents recorded in the presence of neostigmine methyl sulfate, suggesting that the toxin reduced the binding of acetylcholine to endplate receptors. These results are consistent with the hypothesis that conotoxin GI blocks neuromuscular transmission at the frog endplate by reducing the binding of acetylcholine to receptors. PMID- 2981296 TI - Calcium- and cyclic nucleotide-dependent protein kinases and their substrates in the Aplysia nervous system. AB - Homogenates of the Aplysia nervous system contain protein kinase activities sensitive to cAMP, cGMP, and Ca2+/calmodulin. The cAMP- and cGMP-dependent activities are either soluble enzymes or are only loosely bound to membranes, since they can be detected only in crude but not in washed membrane fractions, and are present in 20,000 or 100,000 X g supernatants prepared from homogenates. In contrast there are both soluble and tightly membrane-bound Ca2+/calmodulin dependent protein kinase activities. The three activities present in supernatant fractions can be separated by chromatography on DE-cellulose, indicating that they are due to distinct enzyme species. Substrates for these enzymes were analyzed by two-dimensional gel electrophoresis. Protein phosphorylation within the identified Aplysia neuron R15 in vivo was measured by the intracellular injection of [gamma-32P]ATP. cAMP stimulates the phosphorylation of nine proteins and decreases phosphorylation of two proteins in this cell. This in vivo pattern was compared with in vitro phosphorylation measured in homogenates of whole ganglion. Most of the phosphoproteins affected by cAMP in neuron R15 in vivo are also substrates for cAMP-dependent protein kinase in vitro. Thus, the in vitro system will be a useful tool for detailed biochemical analysis of phosphoproteins which have been identified as being physiologically relevant in vivo. PMID- 2981297 TI - Autoradiographic localization of thyrotropin-releasing hormone receptors in the rat central nervous system. AB - We employed quantitative autoradiography to examine the distribution of thyrotropin-releasing hormone (TRH) receptors in the rat CNS. The binding of [3H]3-methyl-histidine-TRH [( 3H]MeTRH) to TRH receptors in frozen rat brain sections was saturable, of a high affinity (Kd = 5 nM), and specific for TRH analogs. Autoradiograms of [3H]MeTRH binding showed highest concentrations of TRH receptors in the rhinencephalon, including accessory olfactory bulb, nuclei of the amygdala, and the ventral dentate gyrus and subiculum of the hippocampus. Moderate TRH receptor concentrations were found within the thalamus and hypothalamus, in most regions of the rhombencephalon, such as the cranial nerve nuclei, and in the substantia gelatinosa of the spinal cord. Neocortex and basal ganglia contained low densities of TRH receptors. This distribution correlates well with the sensitivity of brain regions to the known effects of TRH, and suggests that TRH receptors may mediate the actions of TRH in the rat CNS. PMID- 2981298 TI - Characterization of an extended glutamate receptor of the on bipolar neuron in the vertebrate retina. AB - The synaptic receptors of ON bipolar neurons are selectively activated by 2-amino 4-phosphonobutyrate, a glutamate analogue. This agent uniquely distinguishes these receptors from other types of excitatory amino acid receptors found in the retina. Various glutamate and aspartate analogues were used to assess the structure-activity characteristics of this receptor. The results suggest that it represents one class of glutamate receptor which can be distinguished by its preferential activation by acidic amino acid analogues that match the extended conformation of glutamate. PMID- 2981299 TI - Corticotropin-releasing factor desensitization of adrenocorticotropic hormone release is augmented by arginine vasopressin. AB - The desensitization of corticotropin-releasing factor (CRF)-stimulated ACTH release from and intracellular cyclic AMP accumulation in anterior pituitary cells was investigated in primary cultures of rat pituitary cells and in a mouse tumor cell line (AtT 20/D16-16). CRF potently stimulated ACTH secretion and cyclic AMP accumulation in both preparations. When primary cultures were preincubated with 100 nM CRF, 50% desensitization of ACTH release occurred within 4 hr while similar desensitization of cyclic AMP accumulation occurred by 1 hr. This desensitization was manifested as a reduced maximal CRF response. Concentrations of CRF as low as 1 to 10 pM induced desensitization. Pretreatment did not affect ACTH content nor did it alter the ability of epinephrine or forskolin to stimulate ACTH release. Pretreatment of AtT 20 cells, which are a homogeneous population of corticotrophs, with 100 nM CRF reduced the subsequent ability of CRF, but not of isoproterenol or of forskolin, to stimulate ACTH release and cyclic AMP formation. Preincubation of AtT 20 cells with isoproterenol did not affect the cyclic AMP or ACTH release responses induced by CRF, but did desensitize beta-adrenergic receptors. Arginine vasopressin (AVP) was a weak ACTH-releasing factor, but AVP enhanced CRF-directed ACTH release from primary cultures. When these cells were preincubated with both CRF and AVP, CRF desensitization occurred with lower concentrations of CRF than when CRF desensitization was induced by preincubating the cells with CRF alone. The ED50 for CRF-induced desensitization was 700 +/- 150 pM when the cells were exposed to CRF alone, and 20 +/- 15 pM when 1 microM AVP was added during the preincubation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981300 TI - Lung tumor imaging by positron emission tomography using C-11 L-methionine. AB - This paper described the first clinical study of lung tumor scanning by positron emission tomography (PET) using C-11-labeled L-methionine (11C-L-Met). Tumor images were clearly visualized by high contrast in eight lung cancer patients and also in a patient with a silicotic nodule. Quantitative evaluations of methionine uptake in tumor tissue and normal tissue by comparing differential uptake ratios suggested that the extent to which 11C-L-Met accumulates in a tumor is closely correlated to the tumor's viability such as benign or malignant, viable or necrotic. 11C-L-Met is considered to be an effective tumor marker for PET diagnosis which represents increased amino acid metabolism of tumors in the mediastinum and lung. PMID- 2981301 TI - Limited significance of asymmetric adrenal visualization on dexamethasone suppression scintigraphy. AB - To assess whether a single measurement of the adrenal uptake of 6 beta-[131I] iodomethylnorcholesterol (NP-59) on constant dexamethasone suppression would allow discrimination of adenoma from normal and bilateral hyperplasia, the adrenal uptake of 6 beta-[131I]iodomethylnorcholesterol (NP-59) was determined in 50 patients with primary aldosteronism (30 adenoma, 20 hyperplasia) and in 13 with hyperandrogenism (six adenoma, seven hyperplasia). Bilateral adrenal NP-59 activity at 5 days was seen in 14 of 36 patients with adenoma (normal to adenoma ratio of greater than or equal to 0.5), whereas marked asymmetric uptake of NP-59 was seen in six of 27 patients with hyperplasia (uptake ratio of less than or equal to 0.5). Thus the level of adrenal NP-59 uptake does not alone serve to distinguish either adenoma from the normal, contralateral adrenal or the adrenal glands in bilateral hyperplasia in all cases. It appears that the pattern of adrenal imaging, early unilateral or early bilateral NP-59 activity (less than 5 days after NP-59 on 4 mg dexamethasone), best serves to separate adrenal adenoma from bilateral hyperplasia. PMID- 2981302 TI - Differential renal function in unilateral renal injury: possible effects of radiopharmaceutical choice. AB - An abnormal filtration fraction or a significant divergence between a kidney's ability to extract Tc-99m dimercaptosuccinic acid (DMSA) and other function parameters, such as the glomerular filtration rate (GFR) or the effective renal plasma flow (ERPF), could lead to different estimates of relative or absolute renal function, depending on the radiopharmaceutical administered. To evaluate this possible divergence, we measured the relative GFR (I-125 iothalamate), ERPF (I-131 hippurate), and Tc-99m DMSA accumulation in adult male Sprague-Dawley rats with unilateral ureteral obstruction or unilateral ischemia at various times after renal injury. The relative ERPF of the obstructed kidney was significantly greater than the relative GFR at all time periods studied; significant but less dramatic differences were noted comparing DMSA with GFR in obstruction and DMSA and ERPF with GFR in ischemia. In evaluating renal disease, it is important to consider the functional parameter reflected by the administered radiopharmaceutical as well as the underlying disease state. PMID- 2981303 TI - Stimulation of absorption of volatile fatty acids and minerals in the cecum of rats adapted to a very high fiber diet. AB - The effects of a high fiber diet containing about 50% pectin, gum, crude potato starch and the fiber components of wheat bran and soya seed cake on the cecal absorption of substrates were studied by parallel measurements of cecal arteriovenous differences and blood flows. Rats fed the high fiber diet had heavier cecae and higher cecal wall weight and blood flow than rats fed a fiber free diet. Very high arteriovenous differences in volatile fatty acids (VFA) were observed and VFA absorption in cecal vein reached 17.5 mumol/min in the high fiber diet group. This process was concomitant to a moderate absorption of Na+ and partly offset by a secretion of Cl-. In contrast, K+, Mg2+ and Ca2+ were absorbed in large amounts along a favorable concentration gradient. About 10% of arterial urea was removed, and there was a substantial reabsorption of ammonia, despite the lower cecal ammonia and the acidic pH in the cecum when the high fiber diet was fed. The present study suggests that rats may tolerate large amounts of diversified fibers or related compounds in the diet. Such a model could contribute to the assessment of the role of VFA in the effect of fiber. PMID- 2981305 TI - Healing of mandibular ridge augmentations using hydroxylapatite with and without autogenous bone in dogs. AB - Dog mandibles were augmented with either hydroxylapatite (HA) alone or HA combined with autogenous bone. Within the limited time frame of this experiment (16 weeks) the group which received HA alone did not demonstrate the induction of bony ingrowth further than 1-2 mm into the augmented ridge. However, the animals augmented with HA combined with autogenous bone demonstrated an active, lamellar type bony ingrowth, maturing bony haversian systems, and vascular elements forming throughout the entire area, extending to the overlying soft tissue interface. The clinical assessment of both types of augmented ridges (HA and HA and bone) were similar; both formed hard, firm ridges at approximately the same rate. This observation is consistent with human clinical experience with HA or HA mixed with bone. However,for patients who have severe mandibular atrophy and for whom increased mandibular bony bulk is crucial, a combination of HA and bone should be advantageous because of the increased extent of bony ingrowth. PMID- 2981304 TI - A modified centripetal approach to parotidectomy. AB - A modified centripetal approach to parotidectomy is reported. A total of 422 benign parotid tumors, 383 of which were primary and 39 of which were recurrent, were treated by this technique. The pleomorphic adenoma was the most common type (94.79%) in the series. Superficial parotidectomy was done in 178 cases, and total parotidectomy, in 244 cases. No permanent facial paralysis occurred after parotidectomy in the primary tumor group. Seven recurrences were observed after surgery in the recurrent tumor group. PMID- 2981306 TI - A method for augmenting the severely atrophic maxilla using hydroxylapatite. PMID- 2981307 TI - Berger disease: Henoch-Schonlein syndrome without the rash. AB - Identical 7-year-old twin boys each had a proved adenovirus infection at the same time. A few days later one developed florid Henoch-Schonlein purpura, severe alimentary tract symptoms, and transient joint symptoms. He had an acute nephritic syndrome, which progressed to nephrotic syndrome and renal insufficiency. Biopsy showed severe proliferative glomerulonephritis with crescents and marked deposition of IgA, IgG, C3, and fibrin. The second twin had hematuria and abdominal pain but no rash. Biopsy showed mesangial proliferative glomerulonephritis with mesangial deposits of IgA and, to a lesser extent, IgG and C3. The appearance was characteristic of Berger disease, and the subsequent clinical course has been that of symptomless microscopic hematuria and recurrent macroscopic hematuria with normal renal function. Immunologic studies have not revealed why these identical twins responded differently to the same provocation. Perhaps Berger disease may be considered a variant of Henoch-Schonlein nephritis. PMID- 2981308 TI - Diabetes insipidus in children. III. Anterior pituitary dysfunction in idiopathic types. AB - Seventeen patients with idiopathic diabetes insipidus occurring in childhood were observed from 4 to 26 years (mean duration 15 1/2 years). The diagnosis of idiopathic diabetes insipidus was based on routine clinical examination and careful, repeated neuroradiologic investigations. Anterior pituitary dysfunction was present in some of these patients. Growth hormone deficiency was present in six children, insufficient thyroid stimulating hormone secretion after thyrotropin-releasing hormone stimulation was demonstrated in one, and abnormal response to a metyrapone test in two. Elevated prolactin and TSH values were present in three and two patients, respectively. Some of these abnormalities were transitory. The presence of anterior pituitary dysfunction in idiopathic diabetes insipidus indicates that the destructive process is not localized to vasopressin synthesizing cells but may also involve other parts of the hypothalamus. PMID- 2981309 TI - Viremia is present in incubation period in nonimmunocompromised children with varicella. PMID- 2981310 TI - Human rotavirus-like particles in a hepatic abscess. PMID- 2981311 TI - Comparison of the effects of diphenylbarbituric acid, phenobarbital, pentobarbital and secobarbital on GABA-mediated inhibition and benzodiazepine binding. AB - We utilized biochemical and electrophysiological methods to compare the effects of anesthetic and anticonvulsant barbiturates on gamma-aminobutyric acid (GABA) receptor-coupled responses. Biochemical analysis involved barbiturate regulation of the affinity of a radiolabeled benzodiazepine for benzodiazepine receptors. The electrophysiological method involved regulation of somatic recurrent inhibition of CA1 pyramidal cells in the hippocampal slice preparation. We found that anesthetic (secobarbital and pentobarbital) but not anticonvulsant (phenobarbital and diphenylbarbituric acid) barbiturates significantly enhanced GABA receptor-coupled responses in both the biochemical and electrophysiological preparations. The results indicate that the anticonvulsant actions of barbiturates may not depend on their ability to enhance GABAergic neuronal activity. PMID- 2981312 TI - Separation of opioid analgesia from respiratory depression: evidence for different receptor mechanisms. AB - Pretreating rats 24 hr earlier with naloxonazine (10 mg/kg i.v.) virtually eliminates the analgesic response observed with morphine at 3.5 mg/kg (i.v.) and significantly reduces the elevation in tail-flick latencies seen with higher morphine doses. Full dose-response curves show a 4-fold shift to the right (P less than .001) following naloxonazine treatment. At 3.5 mg/kg (i.v.), morphine depresses respiratory function, as determined by arterial blood gas (pO2, pCO2 and pH) measurements. Unlike analgesia, prior treatment of rats with naloxonazine does not alter the respiratory depressant actions of morphine. This inability of naloxonazine to antagonize the respiratory depressant actions of morphine is supported by full dose-response curves. Thus, prior treatment of rats with the mu 1-selective antagonist naloxonazine selectively antagonizes analgesia without affecting respiratory depression, implying different receptor mechanisms for the analgesic and respiratory depressant effects of morphine. Further comparisons of the analgesic and respiratory depressant effects of morphine and two opioid peptides, metkephamid and D-Ala2-D-Leu5-enkephalin, strongly suggest the involvement of mu-2 rather than delta mechanisms in opioid respiratory depression. PMID- 2981313 TI - Identification of multiple opiate receptors through neuroendocrine responses. I. Effects of agonists. AB - The effects of the systemic administration of three prototypic multiple opiate receptor agonists, morphine sulfate (MS), ethylketocyclazocine methanesulfonate (EKC) and N-allylnormetazocine hydrochloride (NANMT), on the release of anterior pituitary hormones were studied in the rat. The serum levels of corticosterone, growth hormone, prolactin and luteinizing hormone were measured by radioimmunoassay 30 min after s.c. injection of the drugs. The three opiate compounds elicited different patterns of release of the four hormones. MS, EKC and NANMT elicited rises in the serum levels of corticosterone whereas only MS and EKC induced elevations in growth hormone. MS stimulated but NANMT inhibited the release of prolactin. The administration of the lowest dose of EKC stimulated the release of prolactin whereas higher doses were without effect suggesting biphasic dose-response relationship. The administration of MS, EKC and the highest dose of NANMT elicited a fall in serum luteinizing hormone levels suggesting that NANMT possesses agonist activity at the receptor mediating luteinizing hormone release. The data support the hypothesis that multiple opiate receptors are involved in the mechanism of action of opiate-induced changes in anterior pituitary hormone release. PMID- 2981314 TI - Identification of multiple opiate receptors through neuroendocrine responses. II. Antagonism of mu, kappa and sigma agonists by naloxone and WIN 44,441-3. AB - The effects of the administration of naloxone hydrochloride (NX) or WIN 44,441-3 (WIN), administered either alone or prior to the subsequent administration of the three prototypic multiple opiate receptor agonists morphine sulfate (MS), ethylketocyclazocine methanesulfonate (EKC) and N-allylnormetazocine hydrochloride (NANMT), on the release of anterior pituitary hormones were studied in the rat. Serum levels of corticosterone, growth hormone, prolactin and luteinizing hormone were measured by radioimmunoassay. The administration of WIN alone produced a pattern of hormone release similar to that seen following the administration of NX. Pretreatment with either NX or WIN blocked the MS-induced rise in serum levels of corticosterone whereas neither the EKC nor the NANMT induced rises were blocked. Pretreatment with WIN blocked the EKC-induced rise in serum growth hormone but failed to block the MS-induced increase. Both antagonists blocked the rise in serum levels of prolactin induced by either MS or EKC but also blocked the NANMT-induced fall. The administration of either NX or WIN blocked the inhibition of luteinizing hormone release induced by either MS or NANMT and partially blocked the EKC-induced fall. The data indicate that multiple opiate receptors are involved in opiate-induced changes in anterior pituitary hormone release and suggest that the patterns of hormone release induced by various opiate agonists as well as their interaction with antagonists may be useful in classifying drugs with respect to their activity toward specific receptors. PMID- 2981315 TI - Concentration dependency of the renal vascular and renin secretory responses to adenosine receptor agonists. AB - Exogenous adenosine inhibits renin secretion and can either vasoconstrict or vasodilate the renal vasculature in vivo. In previous experiments, we found that micromolar concentrations of N6-cyclohexyl adenosine and 5'-N-ethylcarboxamide adenosine, analogs which are relatively selective for A1 and A2 adenosine receptors, respectively, tended to have opposite effects on both afferent arteriolar resistance and renin secretory rate in isolated rat kidneys perfused at constant pressure. Because the affinities of A1 and A2 receptors differ considerably, the purpose of the present experiments was to determine the concentration dependencies of the renal hemodynamic (perfusion pressure or perfusate flow, glomerular filtration rate, filtration fraction) and the renin secretory effects of adenosine, N6-cyclohexyl adenosine and 5'-N-ethylcarboxamide adenosine. Both the renovascular and the renin secretory effects of CHA were biphasic; at submicromolar concentrations, total and afferent arteriolar resistances were increased and renin secretion was inhibited. At higher concentrations, these effects were reversed or abolished. 5'-N-Ethylcarboxamide adenosine and adenosine decreased total and segmental (afferent and efferent) resistances and stimulated renin secretion. Taken together, the results are consistent with the suggestions that A1 and A2 receptors mediate inhibition and stimulation of renin secretion, that afferent arteriolar A1 and A2 receptors mediate constriction and dilation and that efferent arteriolar A2 receptors mediate dilation. PMID- 2981316 TI - Cellular mechanisms of drug-induced papillary necrosis. AB - 2-Bromoethylamine hydrobromide (BEA) causes complete papillary necrosis within 24 hr of i.v. administration. The mechanism of this effect is unknown. To characterize further the effect of BEA in transporting epithelia, the urinary bladders of toads and turtles were exposed to varying concentrations of BEA in vitro. In the toad bladder, both cyclic AMP- and vasopressin-stimulated water flow were significantly inhibited after 3 hr of exposure to BEA at a concentration as low as 2.5 X 10(-4) g/ml; after 1 and 2 hr no effect on water transport was observed. Serosal administration of BEA to both toad and turtle bladders significantly inhibited sodium transport to 54% of control at the end of 3 hr. The effect on sodium transport was seen as early as 10 min. The threshold for the effect on sodium transport occurred at a concentration less than that observed for water transport. By contrast, BEA had no effect on hydrogen ion secretion in the isolated turtle bladder over a wide range of concentrations. In fact, after 1 hr, BEA significantly stimulated hydrogen ion secretion. In homogenates of stripped turtle bladder mucosa, BEA significantly inhibited total Na-K adenosine triphosphatase and ouabain sensitive Na-K adenosine triphosphatase. Thus, in anuran membranes, BEA inhibits water and sodium transport but has no effect on acidification. These results suggest that its action in vivo may be related to alterations in cell volume regulation resulting from inhibition of sodium transport rather than a nonspecific toxic effect on the inner medullary epithelium. PMID- 2981317 TI - Muscarinic cholinergic receptors in mouse pituitary tumor cells: prolonged agonist pretreatment decreases receptor content and increases forskolin- and hormone-stimulated cyclic AMP synthesis and adrenocorticotropin secretion. AB - Muscarinic receptor activation on the AtT-20 clonal line of mouse pituitary corticotrophs, inhibits forskolin-stimulated cyclic AMP formation and adrenocorticotropin secretion. In this study, the effect of prolonged receptor stimulation with the muscarinic agonist oxotremorine was found to reduce, in a time-dependent manner, the ability of oxotremorine to inhibit the AtT-20 cell response to forskolin. Pretreatment with oxotremorine also reduced the density of muscarinic receptors without affecting the affinity of these sites for [3H]quinuclidinyl benzilate. In addition to desensitizing the muscarinic receptor, oxotremorine pretreatment also enhanced the ability of forskolin to stimulate cyclic AMP formation and adrenocorticotropin secretion. The apparent sensitizing effect on cyclic AMP synthesis, extended to other muscarinic agents as well as other secretory agonists, was dependent on the oxotremorine concentration used during pretreatment and required at least 2 hr of pretreatment. Enhancement of forskolin-stimulated cyclic AMP accumulation by oxotremorine pretreatment was blocked by cycloheximide and reversed by the muscarinic antagonist, (-)-scopolamine, or by a 5-hr recovery period after pretreatment. The data suggest that prolonged muscarinic receptor activation (rather than simple occupancy) leads to an enhancement of adenylate cyclase activity in AtT-20 cells; whether this effect is coupled to the progressive loss of the inhibitory function of the muscarinic receptor and the receptor down regulation is unknown. PMID- 2981318 TI - Postjunctional alpha-1 and alpha-2 adrenoceptors in the coronaries of the perfused guinea-pig heart. AB - Prazosin and yohimbine were used to differentiate postjunctional alpha adrenoceptors in the coronaries of the perfused guinea-pig heart. Two postjunctional alpha adrenoceptor subtypes were distinguished by the affinities of the receptor for yohimbine and prazosin. The pA2 for yohimbine were 8.74 against alpha-methylnorepinephrine and 8.98 against BHT-920, and the pA2 for prazosin was 9.84 against phenylephrine. Yohimbine was not very active against the alpha-1 selective agonist as was prazosin against the alpha-2 selective agonists. Alpha-1 and alpha-2 postjunctional adrenoceptors mediate vasoconstriction in the whole coronary bed of the perfused guinea-pig heart. PMID- 2981319 TI - Evidence suggesting an Ra-type adenosine receptor in bovine coronary arteries. AB - It has been reported that adenosine has two extracellular membrane receptors depending upon the activation (Ra) or inhibition (Ri) of the adenylate cyclase system. At the Ra site, 5'-N-ethylcarboxamideadenosine is more potent than N6-L phenylisopropyladenosine (L-PIA) and the reverse is true for the Ri site. Therefore, the aim of this investigation was to characterize the subtype of adenosine receptor in bovine coronary arteries with the use of several adenosine analogs. The order of potency for several of these analogs was found to be: 5'-N ethylcarboxamideadenosine greater than 5'-N-cyclopropylcarboxamideadenosine greater than L-PIA greater than N6-cyclohexyladenosine greater than 2-Cl adenosine greater than adenosine greater than D-PIA. The concentration-response curves were parallel to each other, which suggests the same receptor site. L Adenosine, L-5'-N-ethylcarboxamideadenosine, and methyl-N6-cyclohexyladenosine and 2'-5'-dideoxyadenosine (P site analog) were found to be ineffective in relaxing the bovine coronary arteries. The difference between L- and D-PIA was less than 100-fold. This hierarchy suggested the existence of Ra subtype adenosine receptor in bovine coronary arteries. Both theophylline and 8 phenyltheophylline antagonized the relaxing effect of various analogs and shifted the concentration-response curve to the right in parallel. 8-Phenyltheophylline was severalfold more potent in its antagonistic effect than theophylline. In addition, inosine caused the dilation of both large and small coronary arteries at 1 X 10(-4)M and 1 X 10(-3)M concentrations (adenosine would cause a similar relaxation at 500-fold less concentration).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981320 TI - Co-localized adenosine A1 and gamma-aminobutyric acid B (GABAB) receptors of cerebellum may share a common adenylate cyclase catalytic unit. AB - In synaptosomal membranes from rat cerebellum, additive responses to adenylate cyclase activity are observed between the beta adrenergic receptors present on the Purkinje cells and the adenosine A-1 receptors or gamma-aminobutyric acid B (GABAB) receptors, which are both associated with the granule cells. In contrast, nonadditive responses are found with the activation of the adenosine A-1 and the GABAB receptors. Because both receptors are mainly associated with the same cell type, the nonadditive response indicates an interaction between the adenosine A 1, GABAB receptors and the adenylate cyclase. The present study suggests that the nonadditivity results from a limited number of adenylate cyclase catalytic units, which both receptor systems share. This conclusion was derived indirectly by showing that 1) a GABAB agonist did not affect the adenosine A-1 recognition site; 2) both receptors additively activated the high-affinity guanosine 5' triphosphatase, which is believed to reflect the activation of the inhibitory guanine nucleotide unit; and 3) the nonadditivity was still observed after stimulation of adenylate cyclase activity with forskolin. PMID- 2981321 TI - Tissue distribution and functional correlation of [3H]leukotriene C4 and [3H]leukotriene D4 binding sites in guinea-pig uterus and lung preparations. AB - To determine the distribution of leukotriene (LT) C4 and LTD4 receptors and functional significance of each receptor, we used [3H]LTC4 and [3H]LTD4 to measure the binding activity in various tissue homogenates and to correlate the relative ability of the LT agonists to inhibit binding with their contractile responses in guinea-pig uterine or lung parenchymal preparations. Guinea-pig brain contained the highest binding activity of 1 to 2 nM [3H] LTC4 followed by small intestine, heart, lung, kidney, uterus, etc., whereas guinea-pig lung had at least 2.7-fold greater [3H] LTD4 binding activity than any other tissues tested. In the brain and uterine homogenates, the rank order of potency for the compounds in competing with [3H]LTC4 for binding sites was LTC4 much greater than LTD4 greater than LTE4 greater than FPL-55712 = arachidonic acid. The in vitro functional study showed that the ability of the LT agonists to produce uterine contraction was in the order of LTC4 greater than LTD4 greater than LTE4, which is compatible with their relative effect for inhibition of uterine or brain [3H]LTC4 binding. In the lung homogenate, either LTC4, LTD4 or LTE4 inhibited effectively [3H]LTD4 binding and the potency order for the [3H]LTD4 competition study was LTD4 greater than LTE4 greater than LTC4 much greater than FPL-55712 greater than arachidonic acid. These LT agonists also produced lung contraction effectively and the difference among their contractile ability was not significant. We conclude that 1) there are distinct functional LTC4 and LTD4 receptors, 2) activation of the LTC4 receptor could account for the uterine contraction due to the LT agonists and 3) the lung contraction induced by LTC4, LTD4 and LTE4 is at least mediated partly by the LTD4 receptor. PMID- 2981322 TI - Evaluation of the alpha-1 and alpha-2 adrenoceptor-mediated effects of a series of dimethoxy-substituted tolazoline derivatives in the cardiovascular system of the pithed rat. AB - The alpha-1 and alpha-2 adrenoceptor-mediated effects of a series of dimethoxy substituted tolazoline derivatives were investigated in the cardiovascular system of the pithed rat. The 2,5- and 3,5-dimethoxy-substituted tolazoline derivatives produced vasopressor responses that were inhibited by the alpha-1 adrenoceptor antagonist, prazosin (0.1 mg/kg i.v.), and were not affected by the alpha-2 adrenoceptor antagonist, yohimbine (1 mg/kg i.v.), suggesting that these derivatives selectively activate postsynaptic vascular alpha-1 adrenoceptors. The 2,5- and 3,5-dimethoxy-substituted derivatives of tolazoline did not produce an alpha-2 adrenoceptor-mediated inhibition of neurogenic tachycardia in cord stimulated pithed rats and were therefore presumed to be devoid of alpha-2 adrenoceptor agonist activity. In contrast, 2,3-dimethoxytolazoline produced a vasopressor effect that was inhibited by yohimbine but not by prazosin, suggesting selective activation of postsynaptic vascular alpha-2 adrenoceptors. Consistent with this observation is the fact that 2,3-dimethoxytolazoline elicited a dose-dependent, alpha-2 adrenoceptor-mediated inhibition of neurogenic tachycardia in cord-stimulated pithed rat. 3,4-Dimethoxytolazoline was a weak alpha-1 adrenoceptor agonist in the vasculature of the pithed rat and was devoid of agonist activity at alpha-2 adrenoceptors. However, 3,4-dimethoxytolazoline was found to be an alpha-2 adrenoceptor antagonist of similar potency as yohimbine. The results of the present study indicate that dimethoxy-substituted derivatives of tolazoline possess different activities and selectivities at alpha 1 and alpha-2 adrenoceptors depending upon the positions of substitution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981324 TI - Angiotensin-converting enzyme inhibitors. New orally active antihypertensive (mercaptoalkanoyl)- and [(acylthio)alkanoyl]glycine derivatives. AB - A variety of N-substituted (mercaptoalkanoyl)- and [(acylthio)alkanoyl]glycine derivatives was synthesized and their ability in inhibiting the activity of angiotensin-converting enzyme (ACE) was examined in vitro and in vivo. The acylthio derivatives prepared are assumed to act as prodrugs since they are much less active than the corresponding free SH compounds in vitro and can be expected to act in vivo only after conversion to the free sulfhydryl compounds. A number of these compounds are potent ACE inhibitors that lowered blood pressure in Na deficient, conscious spontaneously hypertensive rats (SHR), a high renin model. One of the most active members of the series was (S)-N-cyclopentyl-N-[3-[(2,2 dimethyl-1-oxopropyl)thio]-2-methyl-1 -oxopropyl]glycine (REV 3659-(S), pivopril). Structure-activity relationships are discussed. PMID- 2981323 TI - Uridine 5'-diphosphate glucose analogues. Inhibitors of protein glycosylation that show antiviral activity. AB - A series of analogues of uridine 5'-diphosphate glucose and uridine 5' diphosphate glucosamine have been synthesized by reaction of 2,3,4,6-tetra-O benzyl-, 2,3,4,6-tetra-O-benzoyl-, 2,3,4,6-tetra-O-acetyl-, and 2,3,4,6-tetra-O palmitoyl-alpha-D-glucopyranose and 2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-alpha D-glucopyranose with chlorosulfonyl isocyanate and 2',3'-O-isopropylideneuridine. Isopropylidene and acetyl groups of the resulting 5'-O-[[[[(alpha-D glucopyranosyl)oxy]carbonyl]amino]sulfonyl] -2',3'-O-isopropylideneuridine derivatives were removed by reaction with a TFA/water (5:1) mixture and methanolic ammonia, respectively. The 5'-O-[[[[(2",3",4",6"-tetra-O-benzyl-and 2",3",4",6"-tetra-O-benzoyl-alpha-D-glucopyranosyl)oxy]carbonyl] amino]sulfonyl] 2',3'-O-isopropylideneuridine (13 and 19) and the corresponding deisopropylidenated derivatives showed antiviral activity as determined by the inhibition of the cytopathic effect induced by HSV-1 replication and by the plaque assay method. Compound 13 inhibited glycosylation of proteins in HSV-1 infected HeLa cells. PMID- 2981325 TI - Herpes simplex virus and cytomegalovirus infections in burned patients. AB - Herpesvirus infections are commonly seen in immunosuppressed patients and may account for considerable morbidity and some mortality. We prospectively studied 52 patients with severe burn injuries in order to determine the prevalence of viral infections in this group of patients. Serologic testing was done each week to diagnose primary and reactivation infections. Twenty-seven of 52 patients (52%) became infected with either herpes simplex virus (HSV) or cytomegalovirus (CMV) or both. HSV infection was associated with older age, tracheal intubation, facial burn, inhalation injury, length of hospitalization, and the presence of full-thickness burn. CMV infection was associated with duration of hospitalization and full-thickness burn. Transfusion of blood products was not correlated with an increased incidence of primary or reactivation CMV infections. There was a significant correlation between the presence of these viral infections and bacterial sepsis (p less than 0.05). There was no significant association of HSV or CMV infections with mortality. PMID- 2981326 TI - Differential stability of host mRNAs in Friend erythroleukemia cells infected with herpes simplex virus type 1. AB - The consequences of herpes simplex virus type 1 infection on cellular macromolecules were investigated in Friend erythroleukemia cells. The patterns of protein synthesis, examined by polyacrylamide gel electrophoresis, demonstrated that by 4 h postinfection the synthesis of many host proteins, with the exception of histones, was inhibited. Examination of the steady-state level of histone H3 mRNA by molecular hybridization of total RNA to a cloned mouse histone H3 complementary DNA probe demonstrated that the ratio of histone H3 mRNA to total RNA remained unchanged for the first 4 h postinfection. In contrast, the steady state levels of globin and actin mRNAs decreased progressively at early intervals postinfection. Studies on RNA synthesis in isolated nuclei demonstrated that the transcription of the histone H3 gene was inhibited to approximately the same extent as that of actin gene. We concluded that the stabilization of preexisting histone H3 mRNA was responsible for the persistence of H3 mRNA and histone protein synthesis in herpes simplex virus type 1-infected Friend erythroleukemia cells. The possible mechanisms influencing the differential stability of host mRNAs during the course of productive infection with herpes simplex virus type 1 are discussed. PMID- 2981327 TI - Envelope and long terminal repeat sequences of a cloned infectious NZB xenotropic murine leukemia virus. AB - An infectious NZB xenotropic murine leukemia virus (MuLV) provirus (NZB was molecularly cloned from the Hirt supernatant of NZB-IU-6-infected mink cells, and the nucleotide sequence of its env gene and long terminal repeat (LTR) was determined. The partial nucleotide sequence previously reported for the env gene of NFS-Th-1 xenotropic proviral DNA (Repaske, et al., J. Virol. 46:204-211, 1983) is identical to that of the infectious NZB xenotropic MuLV DNA reported here. Alignment of nucleotide or deduced amino acid sequences, or both, of xenotropic, mink cell focus-forming, and ecotropic MuLV proviral DNAs in the env region identified sequence differences among the three host range classes of C-type MuLVs. Major differences were confined to the 5' half of env; a high degree of homology was found among the three classes of MuLVs in the 3' half of env. Alignment of the nucleotide sequence of the LTR of NZB xenotropic MuLV with those of the LTRs of NFS-Th-1 xenotropic, mink cell focus-forming, and ecotropic MuLVs revealed extensive homology between the LTRs of xenotropic and MCF247 MuLVs. An inserted 6-base-pair repeat 5' to the TATA box was a unique feature of both NZB and NFS-Th-1 xenotropic LTRs. PMID- 2981328 TI - Characterization of a major protein with a molecular weight of 160,000 associated with the viral capsid of Epstein-Barr virus. AB - A monoclonal antibody designated V3 was produced against a late protein associated with the Epstein-Barr virus-induced viral capsid antigen complex. The antibody reacted with discrete patches in the nuclei of infected cells as well as with virus particles, as shown by immunofluorescence and ultrastructural immunoperoxidase staining. The molecular weight of the protein precipitated by this monoclonal antibody was ca. 160,000. All anti-viral capsid antigen antibody positive sera tested in an enzyme-linked immunosorbent assay reacted with this purified protein. The synthesis of the antigen was inhibited by phosphonoacetic acid but was not affected by tunicamycin, indicating that this was a late nonglycosylated viral protein. No differences were noted between the protein isolated from the P3HR-1 and B-95-8 cell lines as determined by immunoprecipitation and peptide mapping. By isoelectric focusing, this protein had a pI on the basic side ranging from 7.5 to 9.0. PMID- 2981329 TI - Middle tumor antigen of polyomavirus transformation-defective mutant NG59 is associated with pp60c-src. AB - We have found that lysis of mouse embryo cells infected with the polyomavirus host range transformation-defective (hr-t) mutant NG59 under gentle conditions that avoid ionic detergents results in detectable NG59-encoded middle tumor antigen (MTAg) associated with pp60c-src. This MTAg-pp60c-src complex could be immunoprecipitated from NG59-infected cell lysates by either sera from animals bearing polyomavirus-induced tumors or by monoclonal antibodies directed against MTAg. Immune complex kinase assays revealed that, whereas the pp60c-src associated with NG59 MTAg possessed tyrosyl kinase activity, the NG59 MTAg in this complex was not phosphorylated in these in vitro reactions. These results demonstrate that the point insertion mutation found in this transformation deficient strain of polyomavirus encodes MTAg molecules capable of associating with pp60c-src and defines a limited region within MTAg which appears to be critical for stable MTAg-pp60c-src interactions. PMID- 2981330 TI - Genetic and biochemical analysis of transformation-competent, replication defective simian virus 40 large T antigen mutants. AB - To study the role of the biochemical and physiological activities of simian virus 40 (SV40) large T antigen in the lytic and transformation processes, we have analyzed DNA replication-defective, transformation-competent T-antigen mutants. Here we describe two such mutants, C8/SV40 and T22/SV40, and also summarize the properties of all of the mutants in this collection. C8/SV40 and T22/SV40 were isolated from C8 and T22 cells (simian cell lines transformed with UV-irradiated SV40). Early regions encoding the defective T antigens were cloned into a plasmid vector to generate pC8 and pT22. The mutations responsible for the defects in viral DNA replication were localized by marker rescue, and subsequent DNA sequencing revealed missense and one nonsense mutation. The T22 mutation predicts a change of histidine to glutamine at residue 203. C8 has two mutations, one predicts lysine224 to glutamamic acid and the other changes the codon for glutamic acid660 to a stop codon; therefore, C8 T antigen lacks the 49 carboxy terminal amino acids. pC8A and pC8B were constructed to contain the C8 mutations separately. Plasmids pT22, pC8, pC8A, and pC8B were able to transform primary rodent cell cultures. T22 T antigen is defective in binding to the SV40 origin. C8B (49-amino-acid truncation) is a host-range mutant defective in a late function in CV-1 but not BSC cells. Analysis of T antigens in mutant SV40 transformed mouse cells suggests that the replicative function of T antigen is important in generating SV40 DNA rearrangements that allow the expression of "100K" variant T antigens in the transformants. PMID- 2981331 TI - Genome organization of herpesvirus aotus type 2. AB - Herpesvirus aotus type 2, a virus commonly found in owl monkeys without overt disease, has a similar genome structure to the oncogenic herpesviruses of nonhuman primates (herpesvirus saimiri, herpesvirus ateles). Virion DNA of herpesvirus aotus type 2 (M-DNA) has an unique 110-kilobase-pair region of low G + C content (40.2%, L-DNA), inserted between stretches of repetitive H-DNA (68.7% G + C, about 41 kilobase pairs per molecule) that are variable in length. A minority of virions contain defective genomes that consist of repetitive H-DNA only. The H-DNA is composed of various types of repeat units that are related in sequence with each other. The two dominant types of repeats (2.3 and 2.7 kilobase pairs) were cloned and compared by restriction enzyme cleavages and partial nucleotide sequencing. They are homologous in at least 1.3 kilobase pairs. The two forms of repeat units are randomly arranged and oriented in tandem. Reassociation kinetics did not allow detection of sequence homologies between H- and L-DNA of herpesvirus aotus type 2 and the respective sequences of oncogenic primate herpesviruses. PMID- 2981332 TI - Evidence for at least two dominant neutralization antigens on human rhinovirus 14. AB - A collection of 28 mutants of human rhinovirus 14, selected for resistance to 10 individual neutralizing monoclonal antibodies, was used to identify two major neutralization antigens, N-Ag I and N-Ag II. Isoelectric analysis showed that all 16 of the N-Ag I mutants analyzed were charge altered in VP1;8 of 12 N-Ag II mutants were altered in VP3. These results suggest that N-Ag I resides on VP1, whereas N-Ag II lies on VP3. The frequency of charge alterations was much higher than predicted by the genetic code, suggesting that charged amino acids on the antigenic sites play an important role in interaction with neutralizing antibody. Antibodies against N-Ag I and N-Ag II neutralize with widely different efficiencies. PMID- 2981333 TI - Accumulation of herpes simplex virus type 1 RNAs of different kinetic classes in the cytoplasm of infected cells. AB - We have analyzed the accumulation of herpes simplex virus type 1 RNA of the immediate early (IE; infected cell polypeptide types 4 and 0 [ICP-4 and ICP-0]), early (thymidine kinase), and early late (ICP-5) kinetic classes in the cytoplasm of infected cells in the presence of anisomycin, canavanine, or phosphonoacetic acid and in the course of a normal infection. IE RNAs were overproduced and were the only class of transcript detected in anisomycin-blocked cells. Phosphonoacetic acid treatment resulted in overaccumulation of early RNAs and underaccumulation of early late RNAs. Although low-stringency canavanine treatment resulted in accumulation of RNA from all kinetic classes, high stringency conditions restricted accumulation of herpes simplex virus type 1 RNAs to the IE class. More importantly, the IE RNAs for ICP-4 and ICP-0 accumulated to a lesser extent under high-stringency canavanine conditions compared with their accumulation in anisomycin-treated cells. Therefore, the absence of newly synthesized viral proteins (anisomysin treatment) and the presence of analog proteins (stringent canavanine treatment) have different consequences with regard to the accumulation of these two IE RNAs. The kinetics of cytoplasmic accumulation for these RNAs was different for each class of RNA. The IE RNAs were detectable at 1 h postinfection and reached a maximum accumulation at ca. 3 h postinfection. The IE RNAs for both ICP-4 and ICP-0 persisted at late times of infection; however, they differed in that the RNA for ICP-4 remained at relatively low levels and the RNA for ICP-0 remained at relatively high levels as compared with their peak levels of accumulation. The 1.4-kilobase RNA for the herpes simplex virus type 1 thymidine kinase was detected by 2 h, with maximum accumulation occurring at ca. 5 h postinfection. After the peak of accumulation, the amount of thymidine kinase RNA declined rapidly from 8 to 14 h postinfection. The early late RNA for ICP-5 was detected between 2 and 3 h, after which accumulation increased to a peak between 8 and 10 h postinfection. The level of ICP-5 RNA remained at close to the peak level until 14 h postinfection. We also compared the accumulation of viral mRNAs in the cytoplasm with the rates of synthesis of their respective polypeptides. Our results suggest that translational controls may be involved in the regulation of IE genes but not early or late genes. PMID- 2981334 TI - Construction of recombinants between molecular clones of murine retrovirus MCF 247 and Akv: determinant of an in vitro host range property that maps in the long terminal repeat. AB - The leukemogenic mink cell focus-forming (MCF) retroviruses such as MCF 247 have biological properties distinct from those of their ecotropic progenitors. Nucleotide sequences encoding portions of gp70, Prp15E, and the long terminal repeat differ between the two types of viruses. To investigate the role of each of these genetic elements in determining the biological properties of MCF viruses, we prepared infectious molecular clones of MCF 247 and generated a set of recombinants between these clones and a molecular clone of Akv, the ecotropic parent of MCF 247. Each molecular clone of MCF 247 was distinct. All the recombinants between Akv and MCF 247 yielded infectious virus upon transfection. Most interestingly, recombinants which contain the long terminal repeat of MCF 247 were found to have an in vitro host range property that has been correlated with high oncogenic activity and thymotropism of certain MCF isolates; namely, they plated with higher efficiency on SC-1 cells than on NFS mouse embryo cells. Nononcogenic MCF isolates showed a slight preference for NFS cells, whereas Akv virus plated with approximately equal efficiency on the two cell types. PMID- 2981335 TI - At least four viral genes contribute to the leukemogenicity of murine retrovirus MCF 247 in AKR mice. AB - Nucleotide sequences encoding gp70, Prp15E, and the U3 region of the long terminal repeat (LTR) distinguish mink cell focus-forming (MCF) retroviruses that can induce leukemia in AKR mice from closely related MCF and ecotropic murine retroviruses that are nonleukemogenic in all inbred mouse strains tested (Lung et al., Cold Spring Harbor Symp. Quant. Biol. 44:1269-1274, 1979; Lung et al., J. Virol. 45:275-290, 1983). We used a set of recombinants constructed in vitro from molecular clones of leukemogenic MCF 247 and nonleukemogenic ecotropic Akv to separate and thereby directly test the role of these genetic elements in disease induction. Leukemogenicity tests of recombinants in AKR mice show that introduction of fragments containing either an MCF LTR or MCF gp70 coding sequences can confer only a very low incidence of disease induction on Akv virus, whereas an MCF type Prp15E alone is completely ineffective. Recombinants with an MCF 247 LTR in combination with MCF Prp15E are moderately oncogenic, whereas those with an MCF 247 LTR plus MCF gp70 coding segment are quite highly leukemogenic. Mice infected with the latter virus show a substantial increase in latent period of disease induction relative to MCF 247; this delay can be reduced when Prp15E, and hence the entire 3' half of the genome, is from MCF 247. Surprisingly, sequences in the 5' half of the genome can also contribute to disease induction. We found a good correlation between oncogenicity and recovery of MCF viruses from thymocytes of injected mice, with early recovery and high titers of MCF in the thymus being correlated with high oncogenicity. This correlation held for recombinants with either an MCF or ecotropic type gp70. Together, these results (i) demonstrate that at least four genes contribute to the oncogenicity of MCF viruses in AKR mice and (ii) suggest that recombinants with only some of the necessary MCF type genes induce leukemia because they recombine to generate complete MCF genomes. Although neither Akv nor MCF 247 is leukemogenic in NFS mice, recombinant viruses whose gp70 gene was derived from Akv but whose LTRs were derived from MCF 247 induced a low incidence of leukemia in this mouse strain. PMID- 2981336 TI - Isolation and structural mapping of a human c-src gene homologous to the transforming gene (v-src) of Rous sarcoma virus. AB - We have utilized a lambda Charon 4A human genomic library to isolate recombinant clones harboring a highly conserved c-src locus containing nucleotide sequences homologous to the transforming gene of Rous sarcoma virus (v-src). Four overlapping clones spanning 24 kilobases of cellular DNA were analyzed by restriction endonuclease mapping. Human c-src sequences homologous to the entire v-src region are present in a 20-kilobase region that contains 11 exons as determined by restriction mapping studies utilizing hybridization to labeled DNA probes representing various subregions of the v-src gene and by preliminary DNA sequencing analyses. A considerable degree of similarity exists between the organization of the human c-src gene and that of the corresponding chicken c-src gene with respect to exon size and number. However, the human c-src locus is larger than the corresponding chicken c-src locus, because many human c-src introns are larger than those of chicken c-src. alu family repetitive sequences are present within several human c-src introns. This locus represents a highly conserved human c-src locus that is detectable in human cellular DNAs from various sources including placenta, HeLa cells, and WI-38 cells. PMID- 2981337 TI - Autoregulation of the bacteriophage P22 scaffolding protein gene. AB - During the formation of each bacteriophage P22 head, about 250 molecules of the product of gene 8, scaffolding protein, coassemble with and dictate correct assembly of the coat protein into a proper shell structure. At approximately the time that DNA is inserted inside the coat protein shell, all of the scaffolding protein molecules leave the structure. They remain active and participate in several subsequent rounds of shell assembly. Previous work has shown that scaffolding protein gene expression is affected by the head assembly process and has generated the hypothesis that unassembled scaffolding protein negatively modulates the expression of its own gene but that it lacks this activity when complexed with coat protein in proheads. To test this model, a P22 restriction fragment containing the scaffolding and coat protein genes was cloned under control of the lac promoter. These cloned genes were then expressed in an in vitro DNA-dependent transcription-translation reaction. The addition of purified scaffolding protein to this reaction resulted in reduced scaffolding protein synthesis relative to coat and tail protein synthesis to an extent and at a protein concentration that was consistent with the observed reduction in vivo. We conclude that scaffolding protein synthesis is autoregulated and that scaffolding protein is the only phage-coded protein required for this process. In addition, these experiments provide additional evidence that this autoregulation is posttranscriptional. PMID- 2981338 TI - A soluble transcription system derived from purified vaccinia virions. AB - A soluble extract from purified vaccinia virus particles has been developed which displays site-specific initiation of transcription on exogenous DNA templates that carry cloned vaccinia virus early gene sequences. Bacterial plasmid vectors with segments of a strongly expressed early region of the vaccinia virus genome were active templates, whether in supercoiled or linear, truncated forms. Correct initiation, corresponding to that found in vivo, was observed for all early genes tested. The involvement of other factors besides the viral RNA polymerase was demonstrated by the loss of specific initiation upon partial purification of the enzyme. Initiation activity was restored by reconstitution of the system with factors lacking polymerase activity. The soluble system retained properties of transcription characteristic of intact viral cores, including (i) similar relative rates of initiation of various genes, (ii) multiple requirement for ATP, (iii) methylation and polyadenylation of transcripts, and (iv) inhibition by a topoisomerase antagonist. PMID- 2981339 TI - A monoclonal antibody specific for a 52,000-molecular-weight human T-cell leukemia virus-associated glycoprotein expressed by infected cells. AB - A monoclonal antibody, designated HT462, is described which is specific for an antigen expressed in human T-cell leukemia/lymphoma virus (HTLV) preparations and by HTLV-infected cells. In indirect immunofluorescence assays, the antigen was detected on the surface of both HTLV-transformed producer and nonproducer cells, including cells infected in vitro with either HTLV subgroup I (HTLV-I) or HTLV II. Normal human peripheral blood lymphocytes stimulated with phytohemagglutinin, cord blood T cells cultured with T-cell growth factor, and a variety of HTLV negative T- and B-cell lines all lacked HT462 antigen expression. The HT462 antigen is a 52,000-molecular-weight glycoprotein, as shown by Western blotting procedures and treatment of viral preparations with neuraminidase, endoglycosidase F, and trypsin. The unglycosylated molecule is approximately 42,000 daltons. That the antigen is virus associated was demonstrated by its banding at the density of HTLV in gradients of metrizamide and by its concomitant synthesis with HTLV gag proteins after short-term culture of primary HTLV positive leukemic cells. Differential expression of the HT462 antigen and HTLV gag-pol gene products was observed. In one case, low HT462 expression was correlated with the known inability of the particular cell line to produce syncytia in vitro. The properties of the HT462 antigen are most consistent with it being a gene product of the HTLV px region or else a cellular antigen specifically induced after viral infection. We cannot rule out, however, that the antigen is a variant cleavage product of the env gene. The monoclonal HT462 will be useful in further definition of the proteins and functions encoded by the env px genetic sequence and in studying the biological properties of HTLV-transformed cells. Furthermore, the monoclonal, by recognizing HTLV-transformed nonproducers, will allow a greater spectrum of virus-infected cells to be detected. PMID- 2981340 TI - Injection of mice with antibody to mouse interferon alpha/beta decreases the level of 2'-5' oligoadenylate synthetase in peritoneal macrophages. AB - Injection of conventional or axenic weanling mice with potent sheep or goat antibody to mouse interferon alpha/beta resulted in a decrease in the basal level of 2-5A synthetase in resting peritoneal macrophages and rendered these cells permissive for vesicular stomatitis virus. There was a good inverse correlation between the level of 2-5A synthetase in peritoneal macrophages and the permissivity of these cells for vesicular stomatitis virus. The peritoneal macrophages of 1- and 2-week-old mice had low levels of 2-5A synthetase and were permissive for vesicular stomatitis virus, whereas at 3 weeks (and after) there was a marked increase in the level of 2-5A synthetase in peritoneal macrophages, and these cells were no longer permissive for vesicular stomatitis virus. We suggest that low levels of interferon alpha or beta or both are produced in normal mice, and that this interferon contributes to host defense by inducing and maintaining an antiviral state in some cells. PMID- 2981341 TI - Protection of mice from wild-type Sendai virus infection by a trypsin-resistant mutant, TR-2. AB - A trypsin-resistant mutant of Sendai virus, TR-2, which could be activated by chymotrypsin but not by trypsin or the protease present in mouse lung, was inoculated intranasally into mice after being activated in vitro. TR-2 hardly brought about clinical illness or lung lesions in mice; the protease present in the lung could not activate the progeny virus, and the infection terminated after one-step replication. Nevertheless, the immunoglobulin A antibody against wild type Sendai virus was produced in the respiratory tracts as well as the serum immunoglobulin G antibody, and the mice were protected from the challenge of the wild-type Sendai virus. On the basis of these results, TR-2 may provide a new model of live vaccine for paramyxoviruses; its availability as a live vaccine is also discussed. PMID- 2981342 TI - In vitro transcription of herpes simplex virus genes: identification of a new initiation site and second intervening sequence in the immediate-early RNA-5 gene. AB - We have used partially purified preparations of RNA polymerase II from mock infected and herpes simplex virus type 1-infected HEp-2 cells to transcribe the herpes simplex virus type 1 strain F BamHI Z DNA fragment containing the promoter for immediate-early RNA-5 (alpha 47), a 1.8-kilobase, spliced RNA. In agreement with the in vivo transcriptional regulation of herpes simplex virus type 1 immediate-early genes, electrophoretic analyses of runoff and truncated transcripts from this template showed that RNA polymerase II from mock-infected cells initiates transcription more selectively than does that from the herpes virus-infected cells at the immediate-early RNA-5 promoter. S1 nuclease mapping of the 5' ends of in vivo- and in vitro-synthesized mRNA placed the initiation site ca. 110 base pairs upstream from the previously published site and also demonstrated the presence of a second, smaller intervening sequence between this new cap site and the previously characterized intervening sequence. S1 analyses also suggested that some splicing of the larger but not the smaller intervening sequence occurred in vitro. PMID- 2981343 TI - Anatomy of the herpes simplex virus 1 strain F glycoprotein B gene: primary sequence and predicted protein structure of the wild type and of monoclonal antibody-resistant mutants. AB - In this paper we report the nucleotide sequence and predicted amino acid sequence of glycoprotein B of herpes simplex virus 1 strain F and the amino acid substitutions in the domains of the glycoprotein B gene of three mutants selected for resistance to monoclonal antibody H126-5 or H233 but not to both. Analyses of the amino acid sequence with respect to hydropathicity and secondary structure yielded a two-dimensional model of the protein. The model predicts an N-terminal, 29-amino-acid cleavable signal sequence, a 696-amino-acid hydrophilic surface domain containing six potential sites for N-linked glycosylation, a 69-amino-acid hydrophobic domain containing three segments traversing the membrane, and a charged 109-amino-acid domain projecting into the cytoplasm and previously shown to marker rescue glycoprotein B syn mutations. The nucleotide sequence of the mutant glycoprotein B DNA fragments previously shown to marker transfer or rescue the mutations revealed that the amino acid substitutions cluster in the hydrophilic surface domain between amino acids 273 and 305. Analyses of the secondary structure of these regions, coupled with the experimentally derived observation that the H126-5- and H233-antibody cognitive sites do not overlap, indicate the approximate locations of the epitopes of these neutralizing, surface reacting, and immune-precipitating monoclonal antibodies. The predicted perturbations in the secondary structure introduced by the amino acid substitutions correlate with the extent of loss of reactivity with monoclonal antibodies in various immunoassays. PMID- 2981344 TI - Constitutive expression of Epstein-Barr virus-encoded RNAs and nuclear antigen during latency and after induction of Epstein-Barr virus replication. AB - We examined the fate of two major products of latency as Epstein-Barr virus was induced to replicate. We studied a superinducible clone of HR-1 cells in the presence and absence of induction by phorbol ester, and we analyzed the X50-7 line with and without superinfection by an HR-1 viral variant which disrupts latency. The two methods of induction yielded qualitatively similar results. After induction, there was abundant synthesis of viral transcripts, amplification of viral DNA, and the appearance of many new viral polypeptides. Nonetheless, there were no changes in the cytoplasmic abundance of Epstein-Barr virus-encoded RNAs and no alteration in the level of Epstein-Barr virus nuclear antigen mRNA or polypeptide. Thus, under conditions in which numerous other Epstein-Barr virus gene products are activated, the two major latent gene products are expressed at a constitutive level. Expression of Epstein-Barr virus-encoded RNAs and nuclear antigen must therefore be regulated in a manner completely different from expression of replicative functions. PMID- 2981345 TI - Analysis of viral and somatic activations of the cHa-ras gene. AB - The activation of the cHa-ras oncogene in the EJ/T24 bladder carcinoma cell line was compared with the activation of the same gene in the rat-derived Harvey murine sarcoma virus. The results indicate that, like the human oncogene, the Harvey murine sarcoma virus-borne ras gene owes its oncogenic capacity to point mutations in coding sequences rather than to the alteration in transcriptional control that occurred when the formerly cellular ras sequences were acquired by the virus. The viral gene retained its transforming ability when its transcription was removed from the influence of the retroviral long terminal repeat promoter and was placed under the regulation of the cHa-ras promoter. Conversely, the viral long terminal repeat was insufficient to activate the normal cHa-ras allele when a single copy of such a construct was delivered to a cell by viral infection. In addition to their mode of activation, the biological properties of the EJ/T24 and Harvey murine sarcoma virus oncogenes were compared by infecting newborn mice with chimeric retroviruses bearing each form. The two alleles acted equivalently, causing erythroleukemias and sarcomas with similar kinetics. PMID- 2981346 TI - Purification of a soluble template-dependent rhinovirus RNA polymerase and its dependence on a host cell protein for viral RNA synthesis. AB - The soluble phase of the cytoplasm of human rhinovirus type 2-infected cells contains an enzymatic activity able to copy rhinovirion RNA without an added primer. This RNA-dependent RNA polymerase (replicase) makes a specific copy of the added rhinovirion RNA, as shown by hybridization of the product to its template RNA but not to other RNAs. The same replicase preparation also contains a virus-specific polyuridylic acid [poly(U)] polymerase activity which is dependent on added polyadenylic acid-oligouridylic acid template-primer. Both activities purify together until a step at which poly(U) polymerase but no replicase activity is recovered. Addition of a purified HeLa cell protein (host factor) to this poly(U) polymerase completely reconstitutes rhinovirus replicase activity. Host factor activity can be supplied by adding oligouridylic acid, suggesting that the host cell protein acts at the initiation step of rhinovirus RNA replication. A virus-specific 64,000-dalton protein purifies with both poly(U) polymerase and replicase activities. PMID- 2981347 TI - Spontaneous expression of C-type virus in DBA/2 mice is associated with an increased rate of mortality, independent of neoplastic disease. AB - Ecotropic C-type retroviruses isolated from both normal and dimethylbenzanthracene-treated DBA/2 mice could be classified into three major groups, Ea, Eb, and Ec, that differed in structure and biology. Weanling DBA/2 mice were generally free of viruses, but a fraction of adult individuals became virus positive and were apparently selectively associated with a high expression of the Eb viruses. Some of the ecotropic viruses from DBA/2 mice acted as exogenous pathogens. They caused viremia and a moderate incidence of leukemia when injected into C3H and ST/a mice. In addition, they caused an appreciable number of early deaths without signs of malignancy. To evaluate the natural role of the viruses, we studied the survival of spontaneously viremic and nonviremic DBA/2 mice. The viremic animals as a group were characterized by a significantly reduced life-span that was not related to neoplasia. These observations indicated that endogenous C-type retroviruses can be pathogenic without preselection of the host for disease. They also emphasize that endogenous viruses, like their exogenous counterparts, can have a broader pathogenic spectrum than normally appreciated. PMID- 2981349 TI - Integration of spleen focus-forming virus proviruses in Friend tumor cells. AB - Using the Southern blot procedure, we studied the presumed spleen focus-forming virus (SFFV) provirus integration sites in the genome of the premalignant and the malignant cells isolated during the course of Friend erythroleukemia. Two restriction endonucleases, PstI and BamHI, discriminated the presumed integrated SFFV proviruses from the endogenous xenotropic-mink cell focus-forming viral sequences. No SFFV integration sites were detectable in the premalignant cells, suggesting a random integration of SFFV proviruses in the genome of these cells. In contrast, SFFV proviruses were detected at a single or very few sites in the genome of all malignant cells we analyzed. These results indicate that the event leading to the malignant transformation in acute Friend leukemia is clonal. In two of the six animals examined, tumors cells isolated from the spleens and the livers of individual mice showed identical SFFV integration patterns. This last result suggests that in some cases different tumors in a same leukemic animal could be derived from a unique clonal event. PMID- 2981350 TI - Involvement of DNA gyrase in bacteriophage T7 growth. AB - We have found that the burst size of bacteriophage T7 was decreased in two Escherichia coli temperature-sensitive gyrase mutants incubated at the restrictive temperature. This reduction in burst size indicates that gyrase may be required for T7 growth. PMID- 2981348 TI - Immunoglobulin subclasses of antibodies to human T-cell leukemia/lymphoma virus I associated antigens in acquired immune deficiency syndrome and lymphadenopathy syndrome. AB - Immunoglobulin G (IgG) and IgM antibodies to human T-cell leukemia/lymphoma virus I (HTLV-I)-associated membrane antigens (HTLV-I-MA) were assayed by indirect cytospin immunofluorescence, and IgG and IgM antibodies to purified HTLV-I were assayed by enzyme-linked immunosorbent assay in sera from 119 immunologically well-characterized promiscuous male homosexuals in The Netherlands, of whom 9 suffered from acquired immune deficiency syndrome (AIDS), 18 suffered from lymphadenopathy syndrome (LAS), and 5 suffered from gay bowel syndrome. Antibodies to HTLV-I-MA were present in four of nine AIDS patients, including one patient with antibodies to purified HTLV-I. Antibodies to HTLV-I-MA were present in 6 of 18 LAS patients, including 3 patients with antibodies to purified HTLV-I. Of five patients with gay bowel syndrome, one had IgG and IgM antibodies to HTLV I-MA. Of the four HTLV-I seropositive AIDS patients, two had IgG and IgM antibodies to HTLV-I or HTLV-I-MA, one had only IgG antibodies, and one had only IgM antibodies. Of the six HTLV-I seropositive LAS patients, four had IgG and IgM antibodies to HTLV-I or HTLV-I-MA, and two had only IgM antibodies. In the sera from 27 healthy homosexuals with and 60 without T-cell subset imbalances, no antibodies to HTLV-I or HTLV-I-MA were detected. PMID- 2981351 TI - Epstein-Barr virus transformation of human B lymphocytes despite inhibition of viral polymerase. AB - Epstein-Barr virus transformed human lymphocytes despite the presence of up to 500 microM acyclovir [9-(2-hydroxyethoxymethyl)guanine], a viral DNA polymerase inhibitor. The transformed cells contained multiple Epstein-Barr virus genome copy numbers. Functional viral DNA polymerase is probably not required for cell transformation and the initial amplification of the viral genome. PMID- 2981352 TI - Naturally occurring BK virus variants (JL and Dik) with deletions in the putative early enhancer-promoter sequences. AB - The genomes of two independently isolated BK virus (BKV) variants (JL and Dik) were compared with prototype BKV DNA by restriction endonuclease mapping and sequence analysis. Differences were mainly detected in two regions: the BKV (JL) and BKV (Dik) putative early enhancer-promoter regions and the middle of the T antigen-coding regions. Base sequence analysis of these two regions showed the following. (i) The putative enhancer-promoter regions of BKV (Dik) and BKV (JL) contained only one 68-base-pair (bp) unit of the 68-bp triplication (the central copy of which is missing 18 bp) present in prototype BKV. (ii) In the same region, BKV (JL) and BKV (Dik) contained unique stretches of DNA 33 and 63 bp long, respectively. In these 63 bp, a sequence which was very similar to the proposed simian virus 40 enhancer core sequence (GGAGTGGAAAG) was present. (iii) The altered restriction endonuclease recognition sites in the sequenced part of the T-antigen-coding region of BKV (JL) and BKV (Dik) were due to base sequence changes, leaving the amino acid sequence unchanged. PMID- 2981354 TI - Transcriptional mapping of the vaccinia virus DNA polymerase gene. AB - Transcriptional analysis of the vaccinia virus DNA polymerase gene revealed the presence of overlapping RNAs. Cloned DNA fragments, previously shown to lie within the DNA polymerase gene (E. V. Jones and B. Moss, J. Virol. 49:72-77, 1984) hybridized to early RNA species of ca. 3,400 and 3,700 nucleotides. Nuclease S1 analysis was used to determine the direction of transcription and more precisely map the mRNAs. A single 5' end and two major and one minor 3' ends were detected. PMID- 2981353 TI - Differences in regulatory sequences of naturally occurring JC virus variants. AB - The regulatory region was sequenced for DNAs representative of seven independent isolates of JC virus, the probable agent of progressive multifocal leukoencephalopathy. The isolates included an oncogenic variant (MAD-4), an antigenic variant (MAD-11), and two different isolates derived from the urine (MAD-7) and from the brain (MAD-8) of the same patient. The representative DNAs were molecularly cloned directly from diseased brain tissue and from human fetal glial cells infected with the corresponding isolated viruses. The regulatory sequences of these DNAs were compared with those of the prototype isolate, MAD-1, sequenced previously (R. J. Frisque, J. Virol. 46:170-176, 1983). We found that the regulatory region of JC viral DNA is highly variable due to complex alterations of the previously described 98-base-pair repeat of MAD-1 DNA. On the basis of these alterations, there are two general types of JC virus. There were no consistent alterations in regulatory sequences which could distinguish brain tissue DNAs from tissue culture DNAs. Furthermore, for each isolate except MAD-1 (R. J. Frisque, J. Virol. 46:170-176, 1983), the regulatory regions of brain tissue and tissue culture DNAs were not identical. The arrangement, sequence, or both of potential regulatory elements (TATA sequence, GGGXGGPuPu, tandem repeats) of JC viral DNAs are sufficiently different from those in other viral and eucaryotic systems that they may effect the unique properties of this slow virus. PMID- 2981355 TI - Mapping of a vaccinia host range sequence by insertion into the viral thymidine kinase gene. AB - A vaccinia virus mutant deleted of ca. 18 kilobase pairs at the left-hand end of the genome is unable to multiply on many human cell lines. To determine whether all or some of the deleted sequences were responsible for the host range property, the corresponding region from wild-type DNA was cloned in three pieces into a vaccinia transplacement vector containing the thymidine kinase gene on the HindIII J fragment. The next step was to transfer these pieces to the genome of the host range deletion mutant by in vivo homologous recombination around the thymidine kinase locus. Transfer of one 5.2-kilobase-pair EcoRI fragment was found to restore a wild-type phenotype on the host range mutant, thus demonstrating that only a small portion of the 18-kilobase-pair deletion contains the host range function(s). This result also illustrates that the method initially devised for inserting foreign genes into vaccinia virus DNA is useful for studies of the vaccinia genome. PMID- 2981356 TI - Biologically active peptides of the vesicular stomatitis virus glycoprotein. AB - A peptide corresponding to the amino-terminal 25 amino acids of the mature vesicular stomatitis virus glycoprotein has recently been shown to be a pH dependent hemolysin. In the present study, we analyzed smaller constituent peptides and found that the hemolytic domain resides within the six amino terminal amino acids. Synthesis of variant peptides indicates that the amino terminal lysine can be replaced by another positively charged amino acid (arginine) but that substitution with glutamic acid results in the total loss of the hemolytic function. Peptide-induced hemolysis was dependent upon buffer conditions and was inhibited when isotonicity was maintained with mannitol, sucrose, or raffinose. In sucrose, all hemolytic peptides were also observed to mediate hemagglutination. The large 25-amino acid peptide is also a pH-dependent cytotoxin for mammalian cells and appears to effect gross changes in cell permeability. Conservation of the amino terminus of vesicular stomatitis virus and rabies virus suggests that the membrane-destabilizing properties of this domain may be important for glycoprotein function. PMID- 2981357 TI - Molecular basis of host range variation in avian retroviruses. AB - Previous genetic analysis has localized the region of the Rous sarcoma virus (RSV) env gene responsible for host range specificity to that encoding the middle one-third of gp85. To better understand the host range determinants, the relevant regions of the genomes of infectious molecular clones of the transformation defective Prague strain of RSV, subgroup B (Pr-RSV-B) and Rous-associated virus 0 (RAV-0) (subgroup E) were sequenced and compared with the sequence of Pr-RSV-C. This comparative analysis identified two variable regions of low amino acid sequence homology flanked by highly conserved amino acid sequences. The first variable region (hr1) begins at base 5654 in the Pr-RSV-C sequence and encodes 32 amino acids. The second variable region (hr2) begins at base 5846 and encodes 27 amino acids. To test the role of the variable regions in host range specificity, we determined the sequence of this region of the env gene of NTRE-4, a recombinant virus between Pr-RSV-B and RAV-0 which exhibits an extended host range. This analysis revealed that the recombinant subgroup-encoding region of NTRE-4 is composed of 200 bases of RAV-0 sequence, including hr2, flanked by sequences which are otherwise of Pr-RSV-B origin. This study indicates that hr1 and hr2 are the domains of gp85 responsible for host range determination in avian retroviruses. PMID- 2981358 TI - Generation of a uniform 3' end RNA of murine leukemia virus. AB - Using the S1 nuclease mapping technique, we demonstrated that the majority of Moloney murine leukemia RNA molecules, isolated either from the nucleus or cytoplasm of infected mouse cells, share a uniform 3' end located at the border of the R and U-5 regions of the long terminal repeat. When the long terminal repeat sequences were inserted in the pSV plasmid downstream of the simian virus 40 late promoter, the 3' end of the viral RNA was also generated close to the R region of the long terminal repeat. These results demonstrate that the long terminal repeat signals the generation of an authentic 3' end when situated downstream of an actively transcribed region. PMID- 2981359 TI - A determinant of polyomavirus virulence enhances virus growth in cells of renal origin. AB - We have identified a strain of polyomavirus, Py(L), which is unusual in causing acute morbidity and early death after inoculation of newborn mice. We determined that these animals died of kidney failure associated with extensive, virus mediated destruction of renal tissue. Interestingly, the Py(L) strain infects baby mouse kidney cell cultures more efficiently than do other strains. PMID- 2981360 TI - Host genetic determinants of neurological disease induced by Cas-Br-M murine leukemia virus. AB - Cas-Br-M is an ecotropic murine leukemia virus (MuLV) of wild-mouse origin that causes neurogenic hind-limb paralysis. By virtue of its N-tropism, the virus replicates well in tissues of mice bearing the n but not the b allele at the Fv-1 locus. To determine if different Fv-1n strains of mice were equally susceptible to virus-induced neurological disease, we inoculated NFS, C3H, DBA/2, CBA, AKR, C58, and NZB mice at birth with Cas-Br-M murine leukemia virus and observed them for the development of tremor and hind-limb paralysis. Three patterns of disease were observed: NFS and C3H mice developed disease within 3 months postinoculation; DBA/2 and CBA mice became affected between 8 and 15 months postinoculation; and no disease was observed in AKR, C58, or NZB mice up to 15 months after infection with Cas-Br-M murine leukemia virus. Studies of genetic crosses between intermediate-latency (DBA/2) or long-latency (AKR) strains with short-latency (NFS) strains showed that intermediate latency and long latency were semidominant traits determined by two or more interacting but independently assorting loci. These genes appear to determine the rate at which the virus replicates and at which viral gene products accumulate in the central nervous system. PMID- 2981361 TI - Assignment of the temperature-sensitive lesion in the replication mutant A1 of vesicular stomatitis virus to the N gene. AB - The replication defect in the temperature-sensitive mutant A1 of the New Jersey serotype (Hazelhurst subtype) of vesicular stomatitis virus was confirmed by the absence of intracellular nucleocapsids in infected cells incubated at the restrictive temperature. After preamplification, the relative yield of the A1 N protein accumulated intracellularly after 1 h of incubation at the restrictive temperature was decreased by 50% that of the wild-type or revertant A1 N protein. This difference was not as apparent in pulse-chase experiments. The functional lesion in A1 was correlated with a structural alteration in the N protein on the basis of the thermolability of the template activity of the A1 N protein-RNA complex in in vitro transcription reactions and the covariance of this phenotype with the temperature-sensitive phenotype in a spontaneous A1 revertant. This correlation was consistent with a direct role of the N protein in replication and allowed the assignment of the N gene to complementation group A. PMID- 2981362 TI - Mapping of the structural gene of pseudorabies virus glycoprotein A and identification of two non-glycosylated precursor polypeptides. AB - Cell-free translation of pseudorabies virus RNA isolated during the late phase of the infectious cycle yielded a variety of polypeptides. A monoclonal antibody directed against one of the major viral glycoproteins, gA, immunoprecipitated two polypeptides ranging in molecular weight from 78K to 83K. To localize the structural gene for gA, we used cloned BamHI fragments of the viral DNA to select specific mRNA species and immunoprecipitated their in vitro translation products with the anti-gA monoclonal antibody. This allowed us to map the genomic region encoding the mRNA for the gA within the short unique region of the viral genome on BamHI fragments 7 and 12. Additional polypeptides encoded by this region were characterized by their electrophoretic mobility. In three virus strains tested a similar, but strain-specific, pattern of the two gA precursors was found which was not dependent on the host cell or the state of infection after reaching the late phase. PMID- 2981363 TI - Myristic acid, a rare fatty acid, is the lipid attached to the transforming protein of Rous sarcoma virus and its cellular homolog. AB - The lipid bound to p60src, the transforming protein of Rous sarcoma virus, has been identified by gas and thin-layer chromatography as the 14-carbon saturated fatty acid, myristic acid. The protein can be labeled biosynthetically with either [3H]myristic acid or [3H]palmitic acid. Incorporation of [3H]myristic acid was noticeably greater than incorporation of [3H]palmitic acid. All of the [3H]myristic acid-derived label in p60src was present as myristic acid. In contrast, none of the radioactivity derived from [3H]palmitic acid was recovered as palmitic acid. Instead, all 3H incorporated into p60src from [3H]palmitic acid arose by metabolism to myristic acid. The cellular tyrosine kinase, p60c-src also contains myristic acid. By comparison of the extent of myristylation of p60v-src with that of the Moloney murine leukemia virus structural protein precursor, Pr65gag, we estimate that greater than 80% of the molecules of p60v-src contain one molecule of this fatty acid. Myristylation is a rare form of protein modification. p60v-src contains 10 to 40% of the myristic acid bound to protein in cells transformed by Rous sarcoma virus and is easily identified in total cell lysates when [3H]myristic acid-labeled proteins are separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparison of the amount of [3H]myristic acid-labeled p60src in total cell lysates and in immunoprecipitates suggests that immunoprecipitation with rabbit anti-Rous sarcoma virus tumor sera detects ca. 25% of the p60src present in cells. PMID- 2981364 TI - Effect of immunosuppression on experimental Argentine hemorrhagic fever in guinea pigs. AB - Immunosuppression with cyclosporin A or cyclophosphamide had no apparent effect on the disease course of guinea pigs infected with a virulent strain of Junin virus. Immunosuppression of guinea pigs infected with an attenuated strain of Junin virus led to fulminating Argentine hemorrhagic fever. All immunosuppressed infected animals died. Virus distribution patterns in target organs, as determined by plaque assay and fluorescent antibody procedures, were similar to those from non-immunosuppressed animals infected with a virulent strain. Histopathological lesions in immunosuppressed guinea pigs infected with an attenuated strain of virus were similar to those in non-immunosuppressed guinea pigs infected with a virulent strain. Histological changes attributable to the immunosuppressive drug(s) were regularly observed. Immunosuppressed animals infected with attenuated Junin virus and non-immunosuppressed animals infected with virulent virus failed to develop antibody or responded at a minimal level. Virus-specific cytotoxic spleen cell activity, previously shown to be antibody dependent, failed to develop in the same animals. The presence of a competent immune response, probably serum antibody, determined whether Argentine hemorrhagic fever infection of the guinea pig was lethal or whether recovery ensued; no evidence for harmful effects of the immune response was obtained. PMID- 2981365 TI - Use of a bacterial expression vector to map the varicella-zoster virus major glycoprotein gene, gC. AB - The genome of varicella-zoster virus (VZV) encodes at least three major glycoprotein genes. Among viral gene products, the gC gene products are the most abundant glycoproteins and induce a substantial humoral immune response (Keller et al., J. Virol. 52:293-297, 1984). We utilized two independent approaches to map the gC gene. Small fragments of randomly digested VZV DNA were inserted into a bacterial expression vector. Bacterial colonies transformed by this vector library were screened serologically for antigen expression with monoclonal antibodies to gC. Hybridization of the plasmid DNA from a gC antigen-positive clone revealed homology to the 3' end of the VZV Us segment. In addition, mRNA from VZV-infected cells was hybrid selected by a set of VZV DNA recombinant plasmids and translated in vitro, and polypeptide products were immunoprecipitated by convalescent zoster serum or by monoclonal antibodies to gC. This analysis revealed that the mRNA encoding a 70,000-dalton polypeptide precipitable by anti-gC antibodies mapped to the HindIII C fragment, which circumscribes the entire Us region. We conclude that the VZV gC glycoprotein gene maps to the 3' end of the Us region and is expressed as a 70,000-dalton primary translational product. These results are consistent with the recently reported DNA sequence of Us (A.J. Davison, EMBO J. 2:2203-2209, 1983). Furthermore, glycosylation appears not to be required for a predominant portion of the antigenicity of gC glycoproteins. We also report the tentative map assignments for eight other VZV primary translational products. PMID- 2981366 TI - Presence of markers for virulence in the unique short region or repeat region or both of pseudorabies hybrid viruses. AB - The unique short region and part of the repeat region of virulent pseudorabies virus strain NIA-3 was replaced by the corresponding region of the avirulent NIA 4 strain by transfection with subgenomic DNA fragments. The resulting hybrid virus showed a reduced virulence in both mice and pigs. Therefore, important markers for virulence are located in the unique short or repeat region or both of pseudorabies virus. We provide evidence that the terminally located repeat is not required for the generation of progeny with intact pseudorabies virus genomes. Apparently, the terminal repeat is regenerated from the internal repeat. PMID- 2981367 TI - Comparison of endogenous murine leukemia virus proviral organization and RNA expression in 3-methylcholanthrene-induced and spontaneous thymic lymphomas in RF and AKR mice. AB - 3-Methylcholanthrene-induced T-cell thymic lymphomas in RF mice were examined for involvement of murine leukemia virus (MuLV)-related sequences in leukemogenesis. Both the expression of MuLV-related RNA species and the organization of endogenous MuLV proviral DNA were analyzed. Of 27 primary tumors examined, only 5 exhibited elevated MuLV-related RNA species homologous to xenotropic specific env DNA. None of these RNA species hybridized with ecotropic p15E DNA sequences. Only two of these five tumors contained MuLV-like RNA species that hybridized with ecotropic MuLV long terminal repeat sequences, despite the probe's ability to detect both ecotropic MuLV and mink cell focus-inducing viral RNA. No muLV resembling mink cell focus-inducing virus whose expression could be correlated with lymphomagenesis was detected in either preleukemic thymocytes, primary 3 methylcholanthrene-induced thymic tumors, tumors passaged in vivo, or cell lines derived from tumors. Restriction endonuclease analysis of DNA from both primary tumors and cell lines failed to reveal either proviral DNA with recombinant env genes or rearrangement of endogenous MuLV proviruses. Therefore, chemically induced lymphomagenesis in RF mice appears different from the spontaneous lymphomagenic process in AKR mice with respect to the involvement of endogenous MuLV sequences. PMID- 2981368 TI - HTLV-III testing of donor blood imminent; complex issues remain. PMID- 2981369 TI - Screening test for HTLV-III (AIDS agent) antibodies. Specificity, sensitivity, and applications. AB - The third member of the human T-cell leukemia (lymphotropic) retrovirus family (HTLV-III) is a newly discovered retrovirus that has been closely associated with the acquired immunodeficiency syndrome (AIDS). In our application of an enzyme linked immunosorbent assay (ELISA) for HTLV-III antibodies, 72 (82%) of 88 patients with AIDS were positive, 14 (16%) were borderline, and two (2%) were negative. In contrast, only 1% of 297 volunteer blood donors were positive, 6% were borderline, and 93% were negative, demonstrating that this ELISA for HTLV III antibodies is highly specific and sensitive for AIDS (excluding borderline results, 98.6% and 97.3%, respectively). Among persons at high risk for AIDS, 8% had borderline results, with positive and negative results readily distinguished as bimodal distributions that paralleled the temporal and geographic trends in AIDS. None of the 188 laboratory and health care employees working with patients with AIDS or their specimens were positive for HTLV-III antibodies, indicating that current precautions for health care workers appear adequate. This ELISA for HTLV-III antibodies will be a useful screening test among blood donors and populations at risk for AIDS, will aid in the diagnosis of suspected AIDS, and will help in defining the spectrum of diseases that are etiologically related to HTLV-III. PMID- 2981370 TI - Perspectives on the future of AIDS. PMID- 2981371 TI - Intracranial lesions in the acquired immunodeficiency syndrome. Radiological (computed tomographic) features. PMID- 2981372 TI - Coexpression of neuroendocrine markers and epithelial cytoskeletal proteins in bronchopulmonary neuroendocrine neoplasms. AB - Neuroendocrine (NE) neoplasms of the human bronchopulmonary tract were examined by electron microscopy, immunocytochemistry, and gel electrophoresis of cytoskeletal proteins from microdissected tissue samples. All samples (carcinoids, well-differentiated NE carcinoma, NE carcinomas of intermediate type, NE carcinomas of the small cell type) contained significant numbers of cells that immunostained for one or more of the following neuroendocrine markers tested: bombesin, calcitonin, ACTH, leu-enkephalin, gastrin, serotonin, somatostatin, alpha-melanocyte-stimulating hormone, vasoactive intestinal peptide, glucagon, insulin, substance P, and neuron-specific enolase. Electron microscopy revealed typical NE cell features, including variable abundant and frequently heterogeneous neurosecretory granules. Tumor cells contained filaments specifically stained with different conventional and monoclonal antibodies to cytokeratins and displayed punctate plasma membrane staining with antibodies to desmoplakins, in agreement with the electron microscopic demonstration of tonofilament bundles and desmosomes. Immunocytochemistry for NE markers and cytoskeletal proteins on consecutive sections revealed both cytokeratins and neuroendocrine substances in single cells. Using gel electrophoresis of cytoskeletal proteins of tissue regions extracted with high salt buffer and detergent, we could detect, in the tumors tested, appreciable amounts of cytokeratin polypeptides 8, 18, and 19, i.e., major cytokeratins also found in certain other lung carcinomas such as adenocarcinomas. Tumor cells were not significantly stained with antibodies to other intermediate filament proteins such as vimentin, desmin, glial filament protein, and neurofilament protein. The results show that NE substances can be synthesized in cells containing a typical epithelial cytoskeleton, i.e., cytokeratin filaments and desmosomes. These findings support the notion of an epithelial character of these tumors and appear in contrast with recent reports that neurofilaments are the only type of intermediate filaments present in carcinoids and other pulmonary NE tumors. These observations may have important implications for the histogenesis of NE carcinomas and for diagnostic pathology. PMID- 2981373 TI - Bronchial carcinoids. Review of 124 cases. AB - A total of 124 patients with bronchial carcinoid were seen at Memorial Sloan Kettering Cancer Center between 1949 and 1983. Of these, 68 were female and 56 were male. The age range was 12 to 82 years (median 55 years). Eleven of the tumors were incidental pathological findings at autopsy or operation and were excluded from survival data determinations. At the time of diagnosis, 82 patients had disease confined to the lung or bronchus, 19 had regional lymph node metastases, and 12 had distant metastases. Patients with distant metastases were more commonly male and smokers, and their tumors were mainly atypical carcinoids histologically, compared with those of patients with localized disease. Patients with distant disease were treated with external radiation and/or chemotherapy, and their median survival was 8 months. Of the 101 patients with disease localized to one hemithorax, endobronchial resection was performed in six and pulmonary resection in 95 (pneumonectomy 14, bilobectomy nine, lobectomy 52, sleeve resection five, segmentectomy 15). Recurrence following endobronchial resection was observed in four of six patients. Disease-free actuarial survival (calculated by the Kaplan-Meier method) following pulmonary resection was 92% at 5 years and 77% at 10 years. Factors predisposing to recurrence were tumor size greater than 3 cm (p less than 0.004), an atypical carcinoid on histologic study (p less than 0.001), and regional lymph node metastases (p = 0.01). Disease-free survival at 5 and 10 years in 19 patients who had regional lymphatic metastases was 74% and 53%, compared with 96% and 84% in those without lymphatic metastases. We conclude that (1) carcinoid tumors are malignant and 10% of patients present with metastases and (2) for patients with clinically localized tumors, the prognosis is determined by the size and histologic features of the tumor and the status of the regional lymph nodes, which must be assessed at thoracotomy. PMID- 2981374 TI - Malignant germ cell tumors of the mediastinum. AB - A review of 56 cases of primary malignant germ cell tumors of the mediastinum revealed that, as with benign teratomas, the tumors occurred in young adults (mean age 29 years) but that the sex distribution differed (86% male and 14% female). A single germ cell element was found in 37 (66%) of the tumors, and various combinations were present in the remaining 19 (34%). The tumors were classified among five recognized types of germ cell tissues. There were 24 seminomas (22 pure and two with mature teratomas), 17 embryonal carcinomas (nine pure and eight with mixtures), five teratomas, seven choriocarcinomas (three pure and four with mixtures), and three pure yolk sac tumors. Most (86%) of the patients were symptomatic at the initial examination, with chest pain, cough, and loss of weight being the most frequent presenting symptoms. The standard posteroanterior and lateral roentgenograms were the most helpful diagnostic tool, showing evidence of an anterior mediastinal mass in 53 patients. The diagnosis was established by surgical exploration of the mediastinum or by biopsy of a lymph node in 55 patients. Of the 55, 24 (43.6%) had complete resection of the tumor and 31 (56.4%) had incomplete resection or biopsy alone. The overall prognosis for mediastinal germ cell tumors is poor, partly because the tumors are far advanced at the time of diagnosis but also because some of the tumors that contain embryonal cell carcinoma, choriocarcinoma, and yolk sac elements are very aggressive. Factors that were prognostic in patients with seminoma--such as age, presence of the superior vena caval syndrome, lymphadenopathy, evidence of hilar disease on the chest roentgenogram, and resectability--were not predictive in patients with other types of malignant germ cell tumors. Although aggressive combination chemotherapy may represent a significant treatment modality for nonseminomatous mediastinal tumors, the present study spanned many years in which no chemotherapy was available. Patients in the later years of the study received combination chemotherapy with various treatment regimens. No conclusions concerning specific chemotherapy, therefore, can be derived from this study. PMID- 2981375 TI - Acrylamide neuropathy in the rat: effects on energy metabolism in sciatic nerve. AB - Energy metabolism was examined in rat sciatic nerve before and after induction of neuropathy by treatment with acrylamide monomer. The in vivo activities of two glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase and nerve-specific enolase, were resistant to acrylamide. The levels of adenosine 5'-triphosphate and creatine phosphate were also unaffected by acrylamide after either short-term or long-term treatment. The turnover of high-energy phosphate was somewhat reduced in the nerves of severely intoxicated animals. These findings cast doubt on the hypothesis that acrylamide neuropathy begins with an attack on the means of generating metabolic energy, although the eventual failure of one or more energy-consuming processes in peripheral nerve remains likely. PMID- 2981376 TI - Toxic neuropathy. PMID- 2981377 TI - Familial euthyroid hyperthyroxinemia secondary to pituitary and peripheral resistance to thyroid hormones. AB - Among 18 family members, representing four generations studied, a familial form of euthyroid hyperthyroxinemia was found in 6. The increased serum total thyroxine value in all hyperthyroxinemic subjects could not be explained by abnormalities in binding proteins. Five of the six patients had a goiter, and all had increased concentrations of triiodothyronine and free thyroxine without symptoms or signs of hyperthyroidism. Basal serum levels of thyroid-stimulating hormone (thyrotropin) were normal in all six; in the four who were tested, these levels responded normally to thyrotropin-releasing hormone (TRH). The normal suppression of basal and TRH-stimulated thyrotropin increase after administration of triiodothyronine did not occur. These patients seem to have resistance of peripheral and pituitary tissues to the actions of thyroid hormones. Family studies revealed that the disorder appeared as a new sporadic mutation and was consistent with an autosomal-dominant mode of inheritance. This disorder, an important example of euthyroid hyperthyroxinemia, should not be confused with Graves' disease. PMID- 2981378 TI - Peripheral receptor populations involved in the regulation of gastrointestinal motility and the pharmacological actions of metoclopramide-like drugs. AB - This minireview is concerned with a re-examination of the locus of action and the possible peripheral mechanisms involved in the gastrointestinal (GI) stimulant effects of metoclopramide. Such a re-evaluation is opportune given the increasing use of this drug in the therapy of certain GI tract disorders. To provide an orientation on this subject the location in the GI tract and function of several relevant receptor types have been reviewed. In the past metoclopramide has been reported to enhance contractions of a variety of GI preparations to electrical stimulation, acetylcholine, carbachol and ganglion stimulants, to inhibit responses to alpha 2-adrenoreceptor agonists and 5-hydroxytryptamine, as well as blocking those to dopamine. Also in such preparations metoclopramide facilitates the release of acetylcholine to transmural stimulation. One important question is whether this effect is mediated via a specific prejunctional receptor. In this respect 2 suggestions have been made. Firstly that the drug may act as a preferential, prejunctional muscarinic antagonist thus inhibiting the negative feedback inhibition of acetylcholine release and secondly that metoclopramide may be a prejunctional agonist (partial) at 5-hydroxy-tryptamine receptors. Although the latter possibility appears most tenable at present, the involvement of a specific receptor remains to be confirmed. The important finding that dopamine receptors are probably not involved in the local stimulant effects of metoclopramide has important implications for future research orientated towards the discovery of a new generation of GI drugs lacking the side effects associated with central dopamine receptor blockade. Several compounds (cinitapride, BRL 20627A and cisapride) are now in the early stages of clinical evaluation. PMID- 2981379 TI - Discriminative stimulus properties of methyl 6,7-dimethoxy-4-ethyl-beta-carboline 3-carboxylate (DMCM), an inverse agonist at benzodiazepine receptors. AB - Rats (N = 8) were trained to discriminate the stimulus properties of the potent benzodiazepine (BZ) receptor inverse agonist methyl 6,7-dimethoxy-4-ethyl-beta carboline-3-carboxylate (DMCM) from saline in a two-lever operant task. The initial training dose of DMCM was 0.4 mg/kg at which the discrimination developed slowly; increasing the dose to 0.8 mg/kg resulted in rapid acquisition. However, since convulsions eventually developed during further training (sensitization), the training dose was finally individualized below the convulsive threshold (0.4 0.7 mg/kg). The DMCM cue was mimicked by FG 7142 (10 mg/kg), a non-convulsant anxiogenic beta-carboline, by pentylenetrazol (20-30 mg/kg), and by the GABA antagonist bicuculline (2 mg/kg). The DMCM cue was not, or marginally, blocked by diazepam (2.5 mg/kg) or pentobarbital (10-15 mg/kg). Furthermore, the BZ receptor antagonists CGS 8216 (2.5 mg/kg), ZK 93426 (20 mg/kg), and Ro 15-1788 (20-80 mg/kg) also did not, or only marginally, block the DMCM cue. However, the receptor antagonists (alone) substituted for DMCM although Ro 15-1788 was less effective. The partial BZ receptor agonist ZK 91296 (25 mg/kg), which is structurally similar to DMCM, blocked completely the DMCM stimulus effect. THIP (4 mg/kg) did not block the DMCM cue. To explain these results, we suggest that the repeated DMCM treatment, necessary for maintaining the discrimination, shifts the balancing point ("set-point") for positive (i.e., BZ-like) agonist efficacy versus inverse agonist efficacy, towards inverse action. This hypothesis was supported by the finding of an enhanced ability of GABA to reduce 3H-DMCM binding to cortical neuronal membranes of animals treated chronically with DMCM in a regimen similar to that used to maintain the DMCM discrimination. Furthermore, this treatment did not affect baseline 3H-DMCM binding, baseline or GABA stimulated 3H-diazepam binding, or 35S-TBPS binding (to chloride channels). PMID- 2981380 TI - Renal and systemic effects of synthetic atrial natriuretic factor. AB - A synthetic peptide corresponding to a sequence of 26 amino acids contained in endogenous rat atrial natriuretic factor (ANF), was infused into one renal artery of anesthetized dogs for a comprehensive in vivo evaluation of the renal and systemic effects of pure ANF. The results proved conclusively that ANF acted directly on the kidney since urine volume and fractional excretion of sodium, potassium, chloride and calcium were elevated in a dose-related manner in the ANF treated kidney, but were not significantly affected in the contralateral saline infused organ. The maximum effects achieved with the synthetic ANF were higher than any reported following intravenous administration of crude extracts of rat atria and were similar to those produced by thiazide diuretics. In four of the five dogs studied, renal vascular resistance fell progressively as doses of ANF were increased. Glomerular filtration rate was not significantly elevated during ANF infusion, but was correlated with sodium excretion rates. Even though mean arterial pressure was progressively reduced, there was no significant change in heart rate and no stimulation of renin secretion. Arterial cyclic GMP concentration was higher in the basal state and rose more rapidly than did renal venous levels, indicating that increases in circulating concentrations of arterial cyclic GMP originated from an extrarenal source. Dose-related elevations in urinary cyclic GMP excretion could be explained by increased cyclic GMP filtration, by enhanced production in tubular cells, or by renal tubular secretion. Especially in the saline-infused kidney, there was a clear dissociation between excretion of cyclic GMP and fractional sodium excretion. We conclude that the synthetic ANF increased electrolyte excretion via a direct renal action which was not solely dependent upon changes in renal vasculature, renin secretion or cyclic GMP levels. PMID- 2981381 TI - Stereospecific opiate receptors in the actions of thyrotropin releasing hormone and morphine on gastrointestinal transit. AB - Studies in these laboratories have shown that morphine and thyrotropin releasing hormone (TRH) inhibit gastrointestinal transit in the mouse. Administration of morphine sulfate (5 mg/kg, s.c.) or TRH (10 microgram i.c.v.) to mice inhibited gastrointestinal transit as measured by the charcoal meal test. In order to determine whether the effects of TRH and morphine were mediated via stereospecific opiate receptors, the effects of two stereoisomers of an antagonist, (-) alpha -5,9-diethyl-2'-hydroxy-2-(3-furylmethyl)6,7-benzomorphan (MR2266), the active isomer and (+) alpha-5,9-diethyl-2'-hydroxy-2-(3 furylmethyl)6,7-benzomorphan (MR 2267), the inactive isomer, on morphine and TRH induced changes in gastrointestinal transit were determined. Morphine and THR induced inhibition of gastrointestinal transit was antagonized by MR 2266 (1 and 3 mg/kg, s.c.) but was unaffected by MR 2267. These studies provide evidence for the involvement of opiate receptors in the actions of morphine and TRH on gastrointestinal transit, and further suggest that the receptors are stereospecific in nature. PMID- 2981382 TI - Mirex-induced liver enlargement in rats is dependent upon an intact pituitary adrenalcortical axis. AB - The effect of prior hypophysectomy upon mirex-induced liver hypertrophy in male Sprague-Dawley rats was examined. Mirex had no effect upon adrenal weight, liver weight, plasma glucose or plasma corticosterone in hypophysectomized rats. However, daily corticosterone supplements (20 mg/kg body weight, sc) given to mirex-treated hypophysectomized animals yielded a 52% increase in liver weight to body weight ratios over those observed in mirex-treated hypophysectomized animals not receiving supplement. In intact rats, both liver weight to body weight ratios and plasma ACTH were significantly increased over controls 2 days after mirex treatment. These results indicate that mirex-induced liver enlargement not only requires corticosterone, but that the response is dependent upon an intact pituitary-adrenalcortical axis. PMID- 2981383 TI - Characterization of angiotensin II binding sites in African Green monkey uterus. AB - The observation that there are significant differences in the concentration, affinity, and specificity of both central nervous system (CNS) and peripheral angiotensin receptors among several different mammalian species, including the African Green monkey, led to the detailed analysis of 125I-angiotensin II binding in the uterus of the African Green monkey. The Bmax for angiotensin receptors in uterine tissue from this species is 56.6 +/- 8.7 fmole per mg protein. The Kd for angiotensin II is .601 +/- .108 nM. The specificity of the receptor is similar to that reported for the uterus of the rat and dog. These results indicate that the angiotensin II receptors, although nearly absent from the CNS of the African Green monkey, are found in the uterus and are very similar to uterine receptors previously characterized in the rat and dog and support the use of these species as appropriate models for studying the biochemistry of angiotensin binding in the uterus. PMID- 2981384 TI - Amygdala kindled seizure stage is related to altered benzodiazepine binding site density. AB - Benzodiazepine receptor binding was examined in rats at 3 stages of amygdaloid kindling (i.e., initial afterdischarge, Stage 3 and Stage 5) immediately or 24 hr after seizure. 3H-diazepam binding site density (Bmax) was significantly increased 24 hr after Stage 3 and Stage 5 kindled seizures in the hippocampus but not in the amygdala. There were no significant differences in the dissociation constants (KD) between kindled and control rats at any time point examined for either brain region. These results demonstrate that changes in benzodiazepine binding are observed with partial kindled seizures (i.e., Stage 3), indicating that generalized seizures are not prerequisite to increased benzodiazepine receptor site density. PMID- 2981385 TI - Characterization of the alpha adrenergic receptor population in hippocampus up regulated by serotonergic raphe deafferentiation. AB - Serotonergic raphe deafferentiation elicits an up regulation of a nM (3H)WB-4101 binding site in rat hippocampus for which norepinephrine displays high affinity and prazosin displays low affinity. Guanine nucleotide affects the nM binding to hippocampal alpha-1 adrenergic receptors. Firstly, Gpp(NH)p, a nonhydrolyzable analog of GTP, inhibits (3H)WB-4101 binding at 3 nM concentration of the radioligand, the ligand concentration labelling the lower affinity, nM, binding site. Secondly, the addition of Gpp(NH)p causes recovery of the heterogeneity of binding sites lost upon preincubation of the membranes with 100 microM epinephrine, apparently by decreasing the affinity of the nM (3H)WB-4101 binding site for the adrenergic receptors. The phenomenon was still observed in the presence of saturating concentrations of the alpha-2 antagonist, yohimbine, and the beta antagonist, propranolol. The results imply that Gpp(NH)p regulates ligand binding to hippocampal alpha-1 agonist sites. It is likely that agonist and antagonist binding sites for the alpha-1 receptor exist in hippocampus with the agonist site being modulated by serotonin. PMID- 2981386 TI - Inhibition of hamster sperm Na+, K+-ATPase activity by taurine and hypotaurine. AB - Taurine, hypotaurine and the structural analogue, beta-alanine, were tested for their effects on Na+, K+-ATPase activity of crude homogenates prepared from washed cauda epididymal hamster sperm. Preincubation with 0.1-10 mM taurine or hypotaurine inhibited Na+, K+-ATPase in a dose-dependent manner, while beta alanine had an inhibitory effect only at 10 mM. The results of this study are the first evidence to demonstrate inhibition of Na+, K+-ATPase activity by taurine and hypotaurine and are discussed in relation to the ability of these compounds to sustain hamster sperm motility and fertility. PMID- 2981387 TI - Quinacrine antagonizes the effects of Na,K-ATPase inhibitors on renal prostaglandin E2 release but not their effects on renin secretion. AB - Previous results have demonstrated that two inhibitors of Na-and-K-activated adenosine triphosphatase (ouabain, vanadate) lead to stimulated prostaglandin E2 release and to inhibited renin secretion in the rat renal cortical slice preparation. It was speculated that stimulation of phospholipase A2 activity accounted for the effect on prostaglandin E2 release. We used the same preparation in the present experiments, and showed that another inhibitor of Na and-K-activated adenosine triphosphatase (K-free incubation medium) stimulates prostaglandin E2 release and inhibits renin secretion. Quinacrine antagonized the stimulatory effects of ouabain, vanadate, and K-free medium on prostaglandin E2 release (consistent with phospholipase A2 involvement), but did not antagonize their inhibitory effects on renin secretion. Collectively, these observations lend further weight to the argument against a mediatory role of prostaglandin synthesis in the renin secretory process. PMID- 2981388 TI - ESR measurement of dissolved oxygen in 4-day chick embryo blood. AB - The magnetic interactions between dissolved oxygen molecules and nitroxide radical spin probes lead to broadening of electron spin resonance (ESR) lines. Based on this property we described an ESR methodology to measure PO2 values in 4 day chick embryo and adult human blood. The total blood volume required for the measurement was only 9 microliter. Using this method PO2 for adult human mixed venous blood was found to be 36.7 mmHg. This is within the established clinical range of 25-40 mmHg for mixed venous blood PO2. The range of mean PO2 values for 4-day chick embryo blood was 27.3- 35.0 mmHg. This is the first time that PO2 values have been reported for individual chick embryos at such an early stage. PMID- 2981389 TI - Changes in the characteristics of low affinity GABA binding sites elicited by Ro15-1788. AB - 3H-GABA binding was studied in cortical membranes from cerebral cortex of handling-habituated and naive rats after the in vitro addition of Ro15-1788. At low concentrations (10(-8), 10(-9) M) Ro15-1788 increased the total number of low affinity 3H-GABA binding sites in brain tissue from naive rats but failed to modify 3H-GABA binding in tissue from handling-habituated ones. On the contrary, Ro15-1788 at higher concentrations (10(-5), 10(-6)M) decreased the total number of low affinity 3H-GABA binding sites in tissue from handling-habituated rats but failed to modify 3H-GABA binding in tissue from naive animals. Ro15-1788 (10( 7)M) failed to modify significantly low affinity 3H-GABA binding in membranes from both naive and handling-habituated rats. However, this concentration abolished the effect of beta-carbolines and diazepam on 3H-GABA binding in membranes from naive and handling-habituated rats, respectively. The changes in the affinity of 3H-GABA binding were inversely related to the changes in the number. The results suggest that: a) the action "in vitro" of Ro15-1788 on low affinity 3H-GABA binding depends from its concentration at the benzodiazepine recognition sites; b) the benzodiazepine recognition site has a modulatory role in the control of the function of GABA-ergic receptor. Our data might explain the conflicting results obtained with this compound "in vivo". PMID- 2981390 TI - Inhibition of growth of C6 astrocytoma cells by inhibitors of calmodulin. AB - We evaluated the effect of several classes of calmodulin inhibitors on the activity of calmodulin prepared from C6 astrocytoma cells and studied the activity of these drugs as inhibitors of the growth of C6 cells in tissue culture. There was a good correlation between the activity of the drugs as inhibitors of calmodulin and their activity as inhibitors of cell growth. The most potent compounds were calmidazolium and melittin as compared to the phenothiazines, trifluoperazine, chlorpromazine, chlorpromazine-sulfoxide or the diphenylbutylpiperidine, pimozide. The mechanism by which the inhibition of calmodulin leads to the death of cells could not be attributed entirely to inhibition of the calmodulin-sensitive cyclic nucleotide phosphodiesterase. Calmodulin is a heat stable, calcium-binding protein involved in numerous biological processes. Recent evidence indicates that calcium and calmodulin may be important for cellular proliferation. For example, this protein changes in concentration during the cell cycle; is involved in the disassembly of the mitotic apparatus; is increased in concentration in rapidly growing hepatomas and in transformed fibroblasts. Weiss and co-workers demonstrated that phenothiazines and structurally similar drugs are capable of binding to and inhibiting the activity of calmodulin. It has been recently observed that certain drugs that inhibit the activity of calmodulin also inhibit the growth of malignant cells in vitro and in vivo. In these studies, however, there was no direct correlation of the effect of the drugs on the calmodulin from the cell type under investigation with cytotoxicity. To learn more about the relationship between a drug's ability to inhibit calmodulin and its antiproliferative activity, we correlated the effect of drugs on the activity of calmodulin prepared from the C6 astrocytoma cell line with their effect on cellular proliferation. Since many inhibitors of calmodulin readily cross the blood-brain barrier and since no acceptable treatment for malignancies of the central nervous system exist, we chose this cell line as a model for elucidating the potential antineoplastic effects of calmodulin inhibitors. PMID- 2981391 TI - Arginine and cysteine residues in the ligand binding site of alpha 2-adrenergic receptors. AB - alpha 2-Adrenergic receptors were identified in calf brain, human platelet and human uterus membranes by [3H]-rauwolscine binding. The reagents phenylglyoxal (selective for guanidino groups), p- hydroxy mercuribenzoate and N-ethylmaleimide (selective for sulfhydryl groups) caused a time- and dose- dependent decrease in the number of receptor sites. alpha 2-Adrenergic agonists and antagonists mediated efficient protection of the receptors against these reagents. These data suggest that essential arginine and cysteine residues are present at or near the alpha 2-adrenergic binding site. PMID- 2981392 TI - Further evidence for the presence of specific binding sites for prolactin in the rabbit brain. Preferential distribution in the hypothalamus and substantia nigra. AB - In the present research we have extended our work on the presence of binding sites for prolactin in the rabbit brain focusing our attention on the brain areas with high dopamine cell bodies density. Among these areas the hypothalamus showed the highest specific binding of labeled ovine prolactin (oPRL). Clearly detectible specific binding was observed also in substantia nigra, whereas in other brain regions the specific binding was very small, except for the striatum where a low but not negligible binding was found in female rabbits. The binding of 125I-oPRL showed a hormonal specificity and Scatchard analysis of the binding showed no clear difference in dissociation constant (Kd) between hypothalamus, nigra and striatum. PMID- 2981394 TI - Provisional Public Health Service inter-agency recommendations for screening donated blood and plasma for antibody to the virus causing acquired immunodeficiency syndrome. PMID- 2981393 TI - Problems in the contemporary treatment of parkinsonism. PMID- 2981395 TI - Respiratory virus surveillance--United States, January 1985. PMID- 2981396 TI - Adverse events following immunization. PMID- 2981397 TI - Alpha 2-adrenergic receptor-mediated regulation of adenylate cyclase in the intact human platelet. Evidence for a receptor reserve. AB - The alpha 2-adrenergic receptor on the human platelet is known to mediate the inhibition of adenylate cyclase activity. A comparison of the binding and response properties of intact cells revealed that the full agonists norepinephrine and epinephrine inhibit cyclic AMP accumulation with apparently higher affinity than they exhibit in inhibiting the binding of [3H]yohimbine. Additionally, Hill coefficients of the occupancy curves of the agonists were less than unity, suggesting the presence of a heterogeneous receptor population in intact platelets under conditions that permit robust inhibition of cyclic AMP accumulation. The partial agonist clonidine was found to possess the same affinity in the binding assay as in the response assay. These data are consistent with the presence of a receptor reserve in this system, a suggestion that was confirmed in experiments utilizing the irreversible alpha 2 antagonist phenoxybenzamine. The IC50 (100 nM) derived from the blockade of [3H]yohimbine binding by phenoxybenzamine was significantly less than the IC50 (550 nM) for the corresponding reversal by phenoxybenzamine of the effects of norepinephrine on cyclic AMP accumulation. Further studies demonstrated a rightward shift in the dose-response curves for the inhibition by norepinephrine of cyclic AMP accumulation following pretreatment with increasing phenoxybenzamine concentration. These data consistently indicated that occupancy of approximately 10% of the alpha 2-adrenergic receptors by norepinephrine elicits a half-maximal adenylate cyclase response. The relationship of these findings to current models of receptor-effector coupling is discussed. PMID- 2981398 TI - Receptors for beta-adrenergic agonists in cultured chick ventricular cells. Relationship between agonist binding and physiologic effect. AB - To determine if the two-state, guanine nucleotide-modulated beta-adrenergic receptor model elucidated in erythrocyte membranes accurately describes hormone binding to intact heart cells, and to determine the relationship of agonist binding to physiologic contractile response, we studied beta-adrenergic antagonist and agonist binding to intact cultured heart cells and homogenates of these cells from embryonic chick ventricle and related the binding observations to alterations in amplitude of contraction of intact cells under identical conditions. The levo isomer of the beta-adrenergic antagonist pindolol was radioiodinated, purified, and utilized to characterize the beta-adrenergic receptor in intact, beating heart cells under physiologic conditions. Computer analysis of iodopindolol-binding isotherms revealed a KD = 22 +/- 3 pM with Bmax = 10.3 fmol/mg of protein in intact cells; in homogenates of cells, the KD was 39 +/- 12 pM in the absence of exogenous guanine nucleotides and 19 +/- 7 pM in their presence. Estimation of the dissociation constant for iodopindolol binding to intact cells by kinetic methods yielded KD = 64 pM. Binding was stereospecific, saturable, and identified a beta 1-adrenergic receptor. Computer modeling of agonist competition curves indicated a single receptor state in intact cells with KD = 0.28 microM for isoproterenol. However, in cell homogenates, two receptor states for agonists were identified with the high affinity state of the receptor having a KD = 3 nM. Addition of guanine nucleotides to the cell homogenate reverted the receptor to a single state similar to that in intact cells. Under nonequilibrium binding conditions, a high affinity state for agonist was detected in intact cells with IC50 = 1.1 nM. The EC50 for isoproterenol-enhanced contractility was 6 nM and EC50 for cAMP response was 4 nM. At the isoproterenol concentration causing 50% maximal inotropic response, 67% occupancy of high affinity receptors occurs. Thus, there is a close relationship between high affinity receptor occupancy and augmentation of contractility in intact cells. These findings support the view that agonist interaction with the guanine nucleotide-sensitive, high affinity receptor state initiates the physiologic response of myocardial tissue to beta-adrenergic agonists. PMID- 2981399 TI - A new approach to determine rates of receptor appearance and disappearance in vivo. Application to agonist-mediated down-regulation of rat renal cortical beta 1- and beta 2-adrenergic receptors. AB - We have developed a method for the assessment of agonist-induced down-regulation of receptors in vivo in terms of rates of receptor appearance and disappearance. This method involves computer-assisted analysis of the kinetics of receptor loss during agonist infusion and of the recovery of receptor number upon the removal of the agonist. These kinetics are analyzed in terms of a steady state model that allows estimation of the rate constants for receptor appearance, kappa ap, and receptor disappearance, kappa dp. Several tests establish that the model can fit experimentally derived data very well. In testing this model, we examined the in vivo down-regulation and recovery of rat renal cortical membrane beta 1- and beta 2-adrenergic receptors in response to infusion of the agonist isoproterenol from subcutaneously implanted osmotic minipumps. During recovery from down-regulation, the beta 1-receptors have a t 1/2 of 45 hr and a kappa ap of 1.6%/hr, and the beta 2-adrenergic receptors a t 1/2 of 18 hr and a kappa ap of 3.9%/hr. During down-regulation, the t 1/2 for both receptors is 12 hr, while kappa ap for beta 1 receptors and beta 2-receptors are 3 and 2.3%/hr, respectively. To the extent that the kinetics of recovery from down-regulation reflect "basal" receptor metabolism, the data indicate that enhanced receptor clearance of both receptor subtypes from the plasma membrane contribute to down-regulation, but changes in rates of receptor appearance may occur as well. The use of this computer modeling technique for defining kinetics of changes in receptor number from one steady state level to another should provide a generally useful means to assess hormone and neurotransmitter receptor metabolism in vivo. PMID- 2981400 TI - Guanine nucleotide-sensitive, high affinity binding of carbachol to muscarinic cholinergic receptors of 1321N1 astrocytoma cells is insensitive to pertussis toxin. AB - Activation of muscarinic cholinergic receptors of 1321N1 human astrocytoma cells attenuates cyclic AMP accumulation. This effect results from an activation of phosphodiesterase with no direct inhibition of adenylate cyclase activity. In spite of this lack of coupling of muscarinic receptors to adenylate cyclase, guanine nucleotides reduce the apparent binding affinity of the agonist carbachol in a washed membrane preparation of 1321N1 cells. The order of potency for this effect is guanosine 5'-O-(3-thiotriphosphate) greater than 5'-guanylyl imidodiphosphate = GTP = GDP; ATP has no effect. The occurrence of a Mr = 41,000 protein labeled in the presence of [32P]NAD and pertussis toxin as well as the occurrence of guanine nucleotide-mediated inhibition of forskolin-stimulated adenylate cyclase activity indicate that the functional inhibitory guanine nucleotide regulatory component of adenylate cyclase (Ni) is present in 1321N1 cells. Pertussis toxin pretreatment of NG108-15 neuroblastoma X glioma cells, which express muscarinic receptors that link through Ni to inhibit adenylate cyclase, blocked the GTP-sensitive, high affinity binding of carbachol. In contrast, pretreatment of 1321N1 cells with a concentration of pertussis toxin that blocked [32P]ADP ribosylation of the Mr = 41,000 substrate and GTP-mediated inhibition of forskolin-stimulated adenylate cyclase activity had no effect on GTP-sensitive high affinity binding of carbachol. These results suggest that muscarinic cholinergic receptors of 1321N1 cells couple to a guanine nucleotide regulatory protein that is distinct from Ni. PMID- 2981401 TI - Characterization of the relationship between gamma-aminobutyric acid B agonists and transmitter-coupled cyclic nucleotide-generating systems in rat brain. AB - Baclofen and other gamma-aminobutyric acid B (GABAB) agonists potentiate the cyclic AMP response in rat brain slices that occurs during exposure to norepinephrine, isoproterenol, adenosine, vasoactive intestinal peptide, and histamine. By themselves the GABAB agonists have only a slight effect on basal cyclic AMP levels. Dose-response and time-course studies revealed that baclofen has little influence on neurotransmitter recognition site affinity, but rather enhances the synthesis or accumulation of second messenger that occurs in response to these agents. Baclofen appears to be neither an inhibitor of phosphodiesterases nor does it require adenosine to promote the response to other transmitters. The synergistic interaction between baclofen and catecholamines is a calcium-dependent process and is evident only in the rat brain cerebral cortex, hippocampus and corpus striatum, being undetectable in the pons-midbrain, cerebellum, and spinal cord. In contrast to the findings with neurotransmitter receptor stimulants, GABAB agonists inhibited the cyclic AMP response to forskolin. It remains unclear whether this action is related to the neurotransmitter potentiating effect of baclofen. These data suggest that GABAB agonists may modulate neurotransmitter receptor function by influencing a component of the cyclic nucleotide-generating system beyond the level of the hormone recognition site. PMID- 2981402 TI - Export of cyclic AMP from avian red cells. Independence from major membrane transporters and specific inhibition by prostaglandin A. AB - Prostaglandin A1 (PGA1) inhibits energy-dependent cyclic AMP export by pigeon red cells [Brunton and Mayer, J. Biol. Chem. 254:9714 (1979)]. To assess the specificity of this action, we observed the effect of 10 microM PGA1 (a concentration that inhibits cyclic AMP efflux greater than 95%) on a variety of membrane-protein-mediated processes that we could readily characterize and quantify in the pigeon red cell. Included in this study were isoproterenol sensitive cyclic AMP production, ouabain-inhibitable 86Rb+ influx, furosemide sensitive NaCl-KCl symport, 4-acetamido-4'-isothiocyano-2, 2'-disulfonic stilbene (SITS)-sensitive sulfate exchange, Na+-dependent alpha-aminoisobutyrate influx, and glucose and adenosine uptake. Remarkably, none of these membrane activities is significantly affected by PGA1. Furthermore, SITS, nitrobenzylthioinosine, cytochalasin B, and Na+-free extracellular medium (inhibitors of band 3, the nucleoside transporter, the hexose transporter, and amino acid uptake, respectively), failed to inhibit cyclic AMP export by pigeon red cells. On the basis of this data, we conclude that PGA1 does not act via a generalized alteration of a basic property at the plasma membrane, such as its fluidity; rather, PGA1 acts specifically to inhibit cyclic AMP extrusion. The data also imply that a transporter not relying on the Na+ gradient and distinct from transporters of cations, anions, amino acids, sugars, and nucleosides mediates cyclic AMP export. PMID- 2981403 TI - Characterization of formylmethionyl-leucyl-phenylalanine stimulation of inositol trisphosphate accumulation in rabbit neutrophils. AB - Inositol trisphosphate (IP3) formed by phospholipase C-mediated breakdown of triphosphoinositide (PIP2) may be a ubiquitous second messenger for a number of Ca2+-mobilizing receptor agonists. Using [3H]inositol-labeled rabbit peritoneal neutrophils, we report that radiolabeled inositol phosphates are generated in response to the chemotactic peptide, formylmethionyl-leucyl-phenylalanine (fMet Leu-Phe). fMet-Leu-Phe-stimulated formation of [3H]IP3 occurs with a rapid time course and a concentration dependence which closely parallels that of stimulated lysosomal enzyme secretion. The synthetic peptide methionyl-leucyl-phenylalanine, which is unable to promote secretion, failed to elevate [3H]IP3 accumulation, and the competitive antagonist t-butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe depressed the stimulant action of fMet-Leu-Phe on [3H]IP3 levels and secretion. The Ca2+ ionophore ionomycin, which promotes secretion, was unable to enhance IP3 levels, confirming that polyphosphoinositide hydrolysis is a specific receptor-mediated event that precedes calcium mobilization during neutrophil activation. The ability of leukotriene B4 to also promote a rapid accumulation of [3H]IP3 suggests that there exists in the neutrophil an interaction between phospholipase A2 and C-mediated events. These findings support the hypothesis that IP3 may be a pivotal messenger for signal transfer by Ca2+-mobilizing receptor agonists. PMID- 2981404 TI - Oxygen-derived free radicals in postischemic tissue injury. AB - It is now clear that oxygen-derived free radicals play an important part in several models of experimentally induced reperfusion injury. Although there are certainly multiple components to clinical ischemic and reperfusion injury, it appears likely that free-radical production may make a major contribution at certain stages in the progression of the injury. The primary source of superoxide in reperfused reoxygenated tissues appears to be the enzyme xanthine oxidase, released during ischemia by a calcium-triggered proteolytic attack on xanthine dehydrogenase. Reperfused tissues are protected in a variety of laboratory models by scavengers of superoxide radicals or hydroxyl radicals or by allopurinol or other inhibitors of xanthine oxidase. Dysfunction induced by free radicals may thus be a major component of ischemic diseases of the heart, bowel, liver, kidney, and brain. PMID- 2981405 TI - Hypotension after quinidine plus verapamil. Possible additive competition at alpha-adrenergic receptors. PMID- 2981406 TI - Circulating activated suppressor T lymphocytes in aplastic anemia. AB - We studied the mechanism of hematopoietic suppression in aplastic anemia by means of two-color flow microfluorometric analysis of lymphocyte subpopulations and correlated the results with the occurrence in vitro of hematopoietic suppression and interferon production. In 12 patients with aplastic anemia a striking increase was observed in a population of "activated" suppressor T lymphocytes, which were defined by binding of both anti-Leu-2 and anti-HLA-DR monoclonal antibodies (patients with aplastic anemia, 6.8 +/- 3.2 per cent [mean +/- S.D.]; normal subjects, 1.7 +/- 1.3; patients given multiple transfusions, 2.5 +/- 1.7). Tac antigen expression, another surface marker of lymphocyte activation, was increased on suppressor lymphocytes in all five patients examined (patients with aplastic anemia, 31 +/- 17 per cent; normal subjects, 0.7 +/- 0.24; patients given multiple transfusions, 2.3 +/- 1.2). When Tac+ and Tac- cells were separated in a cell sorter, only Tac+ cells produced interferon. When lymphocytes of patients with aplastic anemia were cocultured with normal bone marrow, only the Tac+ cell fraction showed hematopoietic suppressor activity. In one patient, in vitro elimination of suppressor lymphocytes by use of OKT8 antibody abolished spontaneous interferon production by bone-marrow cells. These results suggest that activated suppressor lymphocytes producing interferon have a role in the pathogenesis of bone-marrow failure, and indicate the usefulness of defined lymphokine and phenotypic markers in the study of aplastic anemia. PMID- 2981407 TI - Antibodies to HTLV-III in Swiss patients with AIDS and pre-AIDS and in groups at risk for AIDS. AB - We tested serum samples from Swiss subjects by three different assays based on enzyme-linked immunosorbent assay (ELISA) and Western blot techniques for antibodies to proteins associated with the recently discovered human T-cell leukemia/lymphoma virus HTLV-III, the putative etiologic agent for the acquired immunodeficiency syndrome (AIDS). Of 10 patients with AIDS and 10 with pre-AIDS, all were antibody-positive. Furthermore, 37 of 103 intravenous-drug addicts (36 per cent), 4 of 40 healthy homosexual men (10 per cent), 7 of 83 patients with various types of hepatitis (8.4 per cent), but none of 83 healthy blood donors or 10 other controls were antibody-positive. Antibodies to the major viral protein p24 were found consistently and at high titers in the seropositive members of the groups at risk and in those with pre-AIDS but were dramatically reduced in patients with AIDS. In contrast, antibodies to another virus-associated protein, p41, were present in all cases of AIDS and pre-AIDS but were absent in nearly 10 per cent of seropositive persons at risk. Whereas p41 and p24 thus appear to be the targets of choice for future screening tests, the ELISA test that is currently available is a useful screening tool. PMID- 2981408 TI - Hepatitis B virus DNA in patients with chronic liver disease and negative tests for hepatitis B surface antigen. AB - We assessed the presence of hepatitis B virus (HBV) DNA in liver or serum samples from 134 patients with hepatitis B surface antigen (HBsAg)-negative chronic liver disease, including 20 with hepatocellular carcinoma. HBV DNA sequences were detected in 52 of the 88 liver samples (59 per cent), including 17 of the 20 samples from patients with hepatocellular carcinoma. Presumably "replicative forms" of HBV DNA were detected in only 5 of the 88 liver samples, 3 of which were from patients with no serologic marker for HBV. In most of the liver samples the DNA patterns were consistent with the presence of HBV or a closely related virus. Of the 105 serum samples tested, HBV DNA sequences were identified in 10 (9.5 per cent), 6 of which had no HBV serologic marker. Moreover, HBsAg associated determinants were detected in 5 of 17 patients who were positive for HBV DNA and in none of 14 patients who were negative. This study demonstrates the high frequency of HBsAg-negative HBV DNA-positive viral infection of the liver and suggests that multiplication of HBV may occur in the absence of any conventional serologic marker for HBV. PMID- 2981409 TI - Paleolithic nutrition. A consideration of its nature and current implications. PMID- 2981411 TI - Treatment of lymphoproliferation caused by Epstein-Barr virus. PMID- 2981410 TI - Immunologic dysfunction in infants infected through transfusion with HTLV-III. PMID- 2981412 TI - Construction of linkage maps with DNA markers for human chromosomes. AB - DNA markers and sampling of three-generation families can be used to construct complete linkage maps of human chromosomes. This is important in mapping disease loci and in determining the genetic or environmental component of a disease. PMID- 2981413 TI - Amplification, enhanced expression and possible rearrangement of EGF receptor gene in primary human brain tumours of glial origin. AB - Epidermal growth factor (EGF), through interaction with specific cell surface receptors, generates a pleiotropic response that, by a poorly defined mechanism, can induce proliferation of target cells. Subversion of the EGF mitogenic signal through expression of a truncated receptor may be involved in transformation by the avian erythroblastosis virus (AEV) oncogene v-erb-B, suggesting that similar EGF receptor defects may be found in human neoplasias. Overexpression of EGF receptors has been reported on the epidermoid carcinoma cell line A431, in various primary brain tumours and in squamous carcinomas. In A431 cells the receptor gene is amplified. Here we show that 4 of 10 primary brain tumours of glial origin which express levels of EGF receptors that are higher than normal also have amplified EGF receptor genes. Amplified receptor genes were not detected in the other brain tumours examined. Further analysis of EGF receptor defects may show that such altered expression and amplification is a particular feature of certain human tumours. PMID- 2981414 TI - Site-directed mutagenesis of cytochrome c shows that an invariant Phe is not essential for function. AB - Phenylalanine 87 of yeast iso-1-cytochrome c (Phe 82 in horse heart and bonito) is phylogenetically conserved and occurs near the surface of the protein. It has been suggested that this residue is directly involved in electron transfer between cytochrome c and cytochrome c peroxidase (CCP) and may also control the polarity of the haem environment. Because Phe residues are not susceptible to chemical modification, no direct means of studying the functional role of Phe 87 has been available, so we have chosen Phe 87 as our initial target here to test the feasibility of using site-directed mutagenesis as a means of studying structure-function relationships in cytochrome c. We have changed the codon for Phe 87 to that of either a Ser, a Tyr or a Gly. The mutated genes have been introduced into a yeast strain lacking both isozymes of cytochrome c. Unlike the recipient strain, transformants grow on a non-fermentable carbon source, indicating that the mutant proteins can reduce cytochrome oxidase. The purified mutant proteins are similar to wild type with respect to their visible spectra, 20-70% as active as wild-type protein in the CCP assay, and their reduction potentials are lowered by as much as 50 mV. Thus Phe 87 is not essential for cytochrome c to transfer electrons but is involved in determining the reduction potential. PMID- 2981415 TI - DNA bending induced by cruciform formation. AB - Cruciform structures in DNA are of considerable interest, both as extreme examples of sequence-dependent structural heterogeneity and as models for four way junctions such as the Holliday junction of homologous genetic recombination. Cruciforms are of lower thermodynamic stability than regular duplex DNA, and have been observed only in negatively supercoiled molecules, where the unfavourable free energy of formation is offset by the topological relaxation of the torsionally stressed molecule. From an experimental viewpoint this can be a disadvantage, as cruciform structures can be studied only in relatively large supercoiled DNA circles, and are destabilized when a break is introduced at any point. We therefore set out to construct a pseudo-cruciform junction--by generating hereroduplex formation between two inverted repeat sequences. Stereochemically, this should closely resemble a true cruciform but remain stable in a linear DNA fragment. We have now created such a junction and find that it has the expected sensitivities to endonucleases. These DNA fragments exhibit extremely anomalous gel electrophoretic mobility, the extent of which depends on the relative position of the pseudo-cruciform along the length of the molecule. Our results are very similar to those obtained by Wu and Crothers using kinetoplast DNA, and we conclude that the pseudo-cruciform junction introduces a bend in the linear DNA molecule. PMID- 2981416 TI - Differential diagnosis between radiation and tumor plexopathy of the pelvis. AB - We studied 20 patients with lumbosacral radiculoplexopathy from radiation treatment and 30 patients with plexus damage from pelvic malignancy. Indolent leg weakness occurred early in radiation disease, whereas pain marked the onset of tumor plexopathy. Eventually, all radiation cases had weakness, which was bilateral in most of them and painless in one-half of them. Tumor patients typically had unilateral weakness, which was painful in all of them. Radiation disease often resulted in serious neurologic disability. Of the tumor patients, 86% were dead within 3 1/2 years after onset of neurologic symptoms. PMID- 2981417 TI - The natural history of lumbosacral plexopathy in cancer. AB - We studied 85 cancer patients with lumbosacral plexopathy and documented pelvic tumor by CT or biopsy. Three clinical syndromes were delineated: lower (L4-S1), 51%; upper (L1-L4), 31%; and pan-plexopathy (L1-S3), 18%. Seventy percent of patients had the insidious onset of pelvic or radicular leg pain, followed weeks to months later by sensory symptoms and weakness. The quintet of leg pain, weakness, edema, rectal mass, and hydronephrosis suggests plexopathy due to cancer. CT showed pelvic tumor in 96%. On myelography, epidural extension, usually below the conus medullaris, was seen in 45%. With treatment, only 28% of patients had objective responses on CT and 17% on examination. PMID- 2981419 TI - Epithelial ovarian carcinoma of low malignant potential. AB - The records of 94 patients with epithelial ovarian carcinoma of low malignant potential were examined. These records were contributed by Walter Reed Army Medical Center, Naval Hospital, Bethesda, MD, and Naval Hospital, Portsmouth, VA. A review of microscopic sections from each of the 94 tumors confirmed that these were lesions of low malignant potential. The tumors occurred in patients of a younger age than that generally described for invasive epithelial ovarian carcinoma. Forty-seven of 94 patients had stage I disease. The corrected five- and ten-year survival rates were 95 and 87%, respectively. Adjunctive postoperative therapy may not influence survival. PMID- 2981418 TI - Molecular basis of viral neurotropism: experimental reovirus infection. PMID- 2981420 TI - Frostbite. Assessment of tissue viability by scintigraphy. PMID- 2981421 TI - Prejunctional alpha 2-adrenoceptor mediated inhibitory action of clonidine and B HT 920, but not urapidil in guinea pig ileum. AB - Isolated guinea pig ileal longitudinal muscle was stimulated transmurally with a frequency of 0.1 Hz, duration of 0.5 msec, and supramaximal voltage (80-100 V). Transmural stimulation induces ileal contractions via activation of cholinergic neurons. alpha 2-Adrenergic agonists block the response to transmural stimulation via activation of prejunctional alpha 2 receptors which inhibit release of acetylcholine from cholinergic nerve terminals. Urapidil has been reported to have alpha 2-agonistic actions, and therefore was compared to the prototypic alpha 2 agonists, clonidine and B-HT 920. Clonidine and B-HT 920 depressed responses to transmural stimulation in the guinea pig ileum. Clonidine was the most potent inhibitor of the contractions, followed closely by B-HT 920. Very high concentrations of urapidil were necessary to suppress nerve-induced contractions of the ileum. The effects of clonidine and B-HT 920, but not urapidil, were antagonized by the selective alpha 2 antagonist, yohimbine. In unstimulated preparations, in which exogenous acetylcholine was used to elicit contractions of the ileum, urapidil depressed the response while clonidine and B HT 920 had no effect. When PGF1 alpha was used to contract the ileum, no inhibitory effects were noted for urapidil, clonidine, or B-HT 920. Therefore urapidil, only in high concentrations, inhibits the contraction to transmural stimulation by depressing the response at a postjunctional cholinergic site. No evidence was found that urapidil can act as an agonist at a prejunctional alpha 2 receptor site. PMID- 2981422 TI - Phosphatase-mediated modulation of actin-myosin interaction in bovine aortic actomyosin and skinned porcine carotid artery. AB - Since the Ca2+-regulatory mechanism for actin-myosin interaction in smooth muscle involves phosphorylation of the 20,000-Da myosin light chains, it was hypothesized that such interaction should be influenced by myosin phosphatase. Accordingly, we studied the effects of an aortic myosin light-chain phosphatase on Ca1+-dependent actin-myosin interaction in detergent-skinned porcine carotid artery and bovine aortic native actomyosin. In skinned preparations, the aortic phosphatase (16 U/ml) markedly inhibited the rate of isometric contraction in low Ca2+ (6.8 X 10(-7) M) and responsiveness to Ca2+ (force attained with 6.8 X 10( 7) Ca2+/force attained with 1.6 X 10(-6) M Ca2+), whereas relaxation was accelerated. Ca2+-dependent actomyosin ATPase activity and phosphorylation of the light chains were significantly and progressively depressed in the presence of increasing concentrations of phosphatase (0.1-0.9 U/ml). The concentration of Ca2+ (1.1 X 10(-6) M) required for half-maximal activation of either ATPase activity or light-chain phosphorylation increased by 70% in the presence of 0.1 U phosphatase/ml. Neither the maximal rate of Ca2+-sensitive ATP hydrolysis (39 +/- 0.8 nmole/min/mg actomyosin) nor the extent of phosphorylation (0.68 +/- 0.05 mole PO4/mole light chain) was altered at greater than 5 X 10(-6) M Ca2+. ATPase activity was correlated to light-chain phosphorylation under diverse conditions including the presence or absence of 1 microM calmodulin, different concentrations of phosphatase (0-0.9 U/ml), and different concentrations of Ca2+ (10(-8) to 1.25 X 10(-5) M). However, significant phosphorylation was present (20 25% of maximum) in the absence of Ca2+-dependent ATPase activity and only 15% of the maximal rate of ATP hydrolysis was expressed until phosphorylation attained 50% of its maximal value. These findings are consistent with the ordered model of myosin phosphorylation suggested by A. Persechini and D. J. Hartshorne [Science (Washington, DC), 213:1383-285, 1961] (36). They also suggest that myosin phosphatase may participate in modulating actin-myosin interactions in vascular smooth muscle. PMID- 2981423 TI - Effect of malnutrition on rotavirus infection in suckling mice: kinetics of early infection. AB - The effects of malnutrition on the viral replication pattern and severity of clinical disease were examined in suckling mice infected with mouse rotavirus (MRV). The infection in malnourished animals was characterized by a significant decrease in the minimal infectious dose and in the incubation period of the onset of diarrhea, when compared to well-nourished controls. Viral replication in the dispersed enterocytes was observed 6-12 hr earlier, fecal viral shedding peaked significantly earlier, and the clinical disease appeared to be more severe in the malnourished animals than in the controls. These observations provide strong evidence for malnutrition-induced alterations in the pathogenesis of rotaviral infection in vivo. PMID- 2981424 TI - Extracranial tumor vascularity: determination by dynamic CT scanning. Part II: The unit approach. AB - Twenty-eight patients had combined conventional drip infusion CT scans. The information about the anatomic location of the lesion, its configuration, its cross-sectional appearance, its vascularity (as determined by dynamic signature curves), and its clinical presentation were considered as a single overall unit. This diagnostic approach allowed a diagnosis to be made on virtually all of these enhancing lesions without resorting to either a digital venous imaging study or angiographic procedure. In 17 of these cases, such an invasive second procedure was performed either to confirm the CT impression as part of this study or as part of a therapeutic embolization procedure. PMID- 2981425 TI - Pulsed Doppler flow as a criterion of portal venous velocity: comparison with cineangiographic measurements. AB - An ultrasonic sector scanner and pulsed Doppler flowmeter were combined to measure portal venous velocity in 19 patients with portal hypertension and compared with cineangiographic mapping of a droplet of oil released into the portal vein. In 12 patients, measurements were made before and after pitressin [7] or ranitidine [5]. With the Doppler method, maximum basal portal venous velocity was 17.0 +/- 3.9 cm/sec., while average cineangiographic velocity was 8.5 +/- 2.7 cm/sec., a significant difference (p less than 0.001). After pitressin, velocity decreased to 8.3 +/- 2.7 cm/sec. with the Doppler method and 3.6 +/- 1.0 cm/sec. with cineangiography. Ranitidine did not produce an appreciable change. Doppler and cineangiographic velocity measurements exhibited significant correlation over a wide range of values (r = 0.960). Thus the pulsed Doppler method may give accurate values of portal venous velocity if they are corrected to cineangiographic values. Flow can be calculated from velocity and the cross-sectional area of the portal vein as measured on the sonogram. The Doppler method is simple and noninvasive and is particularly useful in studying changes in portal hemodynamics. PMID- 2981426 TI - More progress on the HTLV family. PMID- 2981427 TI - Trans-acting transcriptional regulation of human T-cell leukemia virus type III long terminal repeat. AB - Human T-cell leukemia virus type III (HTLV-III) was recently identified as the probable etiologic agent of the acquired immune deficiency syndrome (AIDS). Here it is shown that, in human T-cell lines infected with HTLV-III, gene expression directed by the long terminal repeat sequence of this virus is stimulated by more than two orders of magnitude compared to matched uninfected cells. The rate of transcription of the HTLV-III long terminal repeat is more than 1000 times that of the SV40 early promoter in one infected cell line. Thus, HTLV-III, like HTLV I, HTLV-II, and the bovine leukemia virus, is characterized by trans-activation of transcription in infected cells. The efficiency of trans-activation in the case of HTLV-III may account, at least in part, for the virulent nature of HTLV III infection. PMID- 2981429 TI - HTLV-III infection in brains of children and adults with AIDS encephalopathy. AB - Unexplained debilitating dementia or encephalopathy occurs frequently in adults and children with the acquired immune deficiency syndrome (AIDS). Brains from 15 individuals with AIDS and encephalopathy were examined by Southern analysis and in situ hybridization for the presence of human T-cell leukemia (lymphotropic) virus type III (HTLV-III), the virus believed to be the causative agent of AIDS. HTLV-III DNA was detected in the brains of five patients, and viral-specific RNA was detected in four of these. In view of these findings and the recent demonstration of morphologic and genetic relatedness between HTLV-III and visna virus, a lentivirus that causes a chronic degenerative neurologic disease in sheep, HTLV-III should be evaluated further as a possible cause of AIDS encephalopathy. PMID- 2981428 TI - Sequence homology and morphologic similarity of HTLV-III and visna virus, a pathogenic lentivirus. AB - A study was conducted of the genetic relation between human T-cell lymphotropic retroviruses and visna virus. The human T-cell lymphotropic viruses include those associated with T-cell malignancies (HTLV-I and HTLV-II) as well as the etiologic agent of the acquired immune deficiency syndrome (HTLV-III). Visna virus, a slowly replicating and pathogenic but nononcogenic retrovirus of sheep, is a member of the subfamily Lentivirinae. Results obtained by molecular hybridization and heteroduplex analysis indicated that a greater extent of nucleotide sequence homology exists between HTLV-III and visna virus than between HTLV-III and any of the other viruses. The homology observed under conditions of low stringency spanned the entire genome, but was strongest in the gag/pol region. The morphogenesis and fine structure of HTLV-III and visna virus also demonstrated striking similarities. The data provide strong evidence for a close taxonomic and thus evolutionary relation between HTLV-III and the Lentivirinae subfamily. PMID- 2981430 TI - Detection of two viral genomes in single cells by double-label hybridization in situ and color microradioautography. AB - Double labeling and color microradioautography were used in a new method of hybridization in situ to identify different genes in individual cells. The method is based on the unequal penetration of 3H and 35S into two layers of nuclear track emulsion separated by a thin barrier film. Hybridization of a 35S-labeled probe specific for one kind of gene results in silver grains over cells in both layers of emulsion; a 3H-labeled probe for a second gene provides grains only in the first layer of emulsion. Silver grains are converted to magenta-colored grains in the first layer and to cyan-colored grains in the second to facilitate enumeration of grains in each layer. This technique should be widely applicable in analyses of differential gene expression in single cells or in discrete populations of cells. PMID- 2981431 TI - Bovine leukemia virus long terminal repeat: a cell type-specific promoter. AB - The functional activity of the promoter unit contained within the long terminal repeat (LTR) of bovine leukemia virus (BLV) was examined by monitoring transient expression of a heterologous gene placed under its control. Various cell lines were transfected with recombinant plasmids carrying the bacterial chloramphenicol acetyltransferase (CAT) gene coupled to the BLV LTR (pBL-cat). Transient expression of CAT activity directed by the BLV LTR was observed only in the established BLV-producer cell lines derived from fetal lamb kidney (FLK) cells and bat lung cells. The amount of CAT activity transiently expressed in FLK-BLV cells was decreased approximately tenfold by deletion of LTR sequences located within a region 100 to 170 nucleotides upstream of the RNA start site. Surprisingly, removal of the region 50 base pairs downstream of the RNA initiation site to the 3'-end of the LTR reduced the expression of CAT activity by 87 percent. The BLV LTR thus appears to be an unusual promoter unit, functioning in a cell type-specific manner and possessing sequences on both the 5' and 3' sides of the RNA start site that influence gene expression. PMID- 2981432 TI - Trans activation of the bovine leukemia virus long terminal repeat in BLV infected cells. AB - The transcription initiation signals for retroviruses lie within the long terminal repeat (LTR) sequences that flank the integrated provirus. This study shows that factors present in cells infected with bovine leukemia virus (BLV) mediate transcriptional trans activation of the BLV LTR. This phenomenon is similar to that reported for the human T-cell leukemia virus LTR and establishes both structural and functional criteria for inclusion of BLV and human T-cell leukemia viruses in the same family of transforming retroviruses. PMID- 2981433 TI - Prooxidant states and tumor promotion. AB - There is convincing evidence that cellular prooxidant states--that is, increased concentrations of active oxygen and organic peroxides and radicals--can promote initiated cells to neoplastic growth. Prooxidant states can be caused by different classes of agents, including hyperbaric oxygen, radiation, xenobiotic metabolites and Fenton-type reagents, modulators of the cytochrome P-450 electron transport chain, peroxisome proliferators, inhibitors of the antioxidant defense, and membrane-active agents. Many of these agents are promoters or complete carcinogens. They cause chromosomal damage by indirect action, but the role of this damage in carcinogenesis remains unclear. Prooxidant states can be prevented or suppressed by the enzymes of the cellular antioxidant defense and low molecular weight scavenger molecules, and many antioxidants are antipromoters and anticarcinogens. Finally, prooxidant states may modulate the expression of a family of prooxidant genes, which are related to cell growth and differentiation, by inducing alterations in DNA structure or by epigenetic mechanisms, for example, by polyadenosine diphosphate-ribosylation of chromosomal proteins. PMID- 2981434 TI - Antibodies to peptides detect new hepatitis B antigen: serological correlation with hepatocellular carcinoma. AB - The expression of a previously unidentified gene product, encoded by the hepatitis B virus (HBV) genome, has been achieved with a recombinant SV40 expression vector. Antibodies against synthetic peptides representing defined regions of this protein were used to screen cells infected with recombinant virus as well as tissues naturally infected with HBV. A 24,000-dalton protein (p24) was detected in cells infected with recombinant virus and a 28,000-dalton protein (p28) was detected in tissues infected with HBV. The peptides or recombinant derived protein were used as antigens to screen sera from individuals infected with HBV. Specific antibodies were detected predominantly in sera from patients with hepatocellular carcinoma. The presence of p28 in tissues infected with HBV and the appearance of specific antibodies in infectious sera establish the existence of an additional marker for HBV infection. PMID- 2981436 TI - AIDS virus genomes. PMID- 2981435 TI - Vaccinia virus recombinants: expression of VSV genes and protective immunization of mice and cattle. AB - Vesicular stomatitis virus (VSV) causes a contagious disease of horses, cattle, and pigs. When DNA copies of messenger RNA's for the G or N proteins of VSV were linked to a vaccinia virus promoter and inserted into the vaccinia genome, the recombinants retained infectivity and synthesized VSV polypeptides. After intradermal vaccination with live recombinant virus expressing the G protein, mice produced VSV-neutralizing antibodies and were protected against lethal encephalitis upon intravenous challenge with VSV. In cattle, the degree of protection against intradermalingually injected VSV was correlated with the level of neutralizing antibody produced following vaccination. PMID- 2981437 TI - Magnetic resonance imaging of biological specimens by electron paramagnetic resonance of nitroxide spin labels. AB - Electron paramagnetic resonance imaging was demonstrated on two plant species, Apium graveolens and Coleus blumei. This was accomplished by soaking stems of these plants in the paramagnetic nitroxide imaging agent 4-hydroxy-2,2,6,6 tetramethylpiperidine-1-oxyl. The experiments were accomplished at L-band frequency (1.4 to 1.9 gigahertz) with single-turn, flat-loop surface coils. One dimensional imaging spectra were diagnostic of capillary structure and long-term stability. PMID- 2981438 TI - Characterization of long terminal repeat sequences of HTLV-III. AB - The nucleotide sequence of the long terminal repeat sequence (LTR) of the human T cell leukemia (lymphotropic) virus type III (HTLV-III) was determined. This virus is associated etiologically with the acquired immune deficiency syndrome. The LTR was found to be 634 base pairs in length with U3, R, and U5 regions of 453, 98, and 83 bp, respectively. The proviral DNA is flanked by a 7-base-pair direct repeat. The promoter and polyadenylation signals are situated 27 and 24 base pairs upstream from the respective transcriptional initiation and polyadenylation sites. The primer binding site is complementary to transfer RNA-lysine. The LTR of HTLV-III, like that of HTLV-I, showed a limited homology to enhancer-like sequences within two genes expressed specifically in T lymphocytes, T-cell growth factor, and gamma-interferon. Structural comparisons revealed that the LTR of HTLV-III is distantly related to those of HTLV-I, HTLV-II, and bovine leukemia virus. PMID- 2981439 TI - Hypothalamic reward mechanism: two first-stage fiber populations with a cholinergic component. AB - Refractory periods were estimated for fibers of the hypothalamic substrate of brain stimulation reward. Two nonoverlapping populations were evident: a homogeneous fast population and a more heterogeneous slow population. Cholinergic blockade eliminated the contribution of the fast but not the slow fibers, while dopaminergic blockade reduced responding without significantly influencing either directly activated fiber population. These data indicate that the hypothalamic reward substrate is more complex than has been widely appreciated; it contains two or more parallel subsystems, and one of these subsystems has a cholinergic link. PMID- 2981440 TI - Intraoperative hepatic ultrasonography--an indispensable procedure in resection of small hepatocellular carcinomas. AB - Hepatic ultrasonography (US) was performed during laparotomy in 47 patients with hepatocellular carcinomas (HCC) less than 5 cm in size. It detected more tumors than did preoperative US and other imaging modalities. In this series, 45.9% of HCCs smaller than 3 cm and 14.2% of those between 3 and 5 cm were invisible and impalpable during laparotomy, and another 15.5% of the total of HCCs were only partially visible or equivocally palpable; thus they needed intraoperative US to make a three-dimensional localization of the tumors. We concluded that, in the resection of small HCCs, intraoperative US should serve as a routine procedure. PMID- 2981442 TI - Cyclosporine-associated central nervous system toxicity after renal transplantation. PMID- 2981441 TI - Alterations in alpha adrenergic receptors in human cerebral arteries after subarachnoid hemorrhage. AB - The nature of alpha adrenergic receptors in human cerebral arteries was characterized and alteration of these receptors after subarachnoid hemorrhage (SAH) was examined using a radioligand binding assay. The specific 3H-prazosin binding to human cerebral arteries was saturable and of high affinity (KD = 4.1 nM) with a Bmax of 92 fmol/mg protein. Specific 3H-yohimbine binding to these tissues was also saturable and of high affinity (KD = 23 nM) with a Bmax 250 fmol/mg protein. IC50 values of adrenergic agents for 3H-prazosin binding were as follows: prazosin, 1.2 X 10(-10) M; phentolamine, 1.3 x 10(-6) M; yohimbine, 1.2 x 10(-5); norepinephrine, 4.9 x 10(-4) M; epinephrine greater than 1 x 10(-3) M. IC50 values of adrenergic agents for 3H-yohimbine binding were as follows: phentolamine, 1.7 x 10(-7) M; yohimbine, 4.2 x 10(-7) M; prazosin, 1.9 x 10(-5) M; epinephrine, 4.4 x 10(-5) M; norepinephrine, 7.9 x 10(-4) M. KD and Bmax of 3H prazosin and 3H-yohimbine binding after SAH were compared with findings in the non-SAH group. KD and Bmax of 3H-prazosin binding of SAH group were 6 +/- 3 nM and 90 +/- 10 fmol/mg protein, respectively (N = 3). KD and Bmax of 3H-yohimbine binding of SAH group were 42 +/- 6 nM and 460 +/- 30 fmol/mg protein, respectively (N = 5). On the other hand, KD and Bmax of 3H-prazosin binding in the non-SAH group were 4 +/- 1 nM and 90 +/- 20 fmol/mg protein, respectively (N = 5), KD and Bmax of 3H-yohimbine binding of non-SAH group were 20 +/- 5 nM and 260 +/- 30 fmol/mg protein, respectively (N = 6).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981444 TI - Oat cell carcinoma of lung masquerading as prostatic carcinoma. AB - We believe this is the second case of oat cell carcinoma metastatic to the prostate to be reported. The patient was treated for prostatic carcinoma diagnosed by rectal examination. Subsequent biopsy confirmed oat cell carcinoma and led to the discovery of pulmonary carcinoma. Tumors metastatic to the prostate are rare and are identified when they result in urologic symptoms. A review of tumors metastatic to the prostate is presented. PMID- 2981443 TI - Late-onset interstitial pneumonia following allogeneic bone marrow transplantation. AB - Although most episodes of interstitial pneumonia (IP) following allogeneic bone marrow transplantation occur during the second or third month, occasional cases occur later. The records of 139 patients transplanted for leukemia over 6 years were reviewed in this study to examine cases of IP occurring beyond 100 days following marrow transplantation. Ten episodes of IP occurred among the 67 patients who survived 100 days (an actuarial probability of 18%). All cases occurred within the first year. An infectious cause was established in 80%. The case fatality rate was 60%. Although most infectious cases of IP occurring before 100 days are caused by cytomegalovirus, the late-onset cases in this study were caused by a heterogeneous group of pathogens, some of which were amenable to specific therapies. Thus, an aggressive approach to establishing a specific diagnosis should be made in cases of IP occurring more than 100 days after marrow transplantation. PMID- 2981446 TI - Tumorigenic poxviruses: genomic organization of malignant rabbit virus, a recombinant between Shope fibroma virus and myxoma virus. AB - The genome of malignant rabbit virus (MRV), a newly discovered tumorigenic poxvirus of rabbits, has been analyzed using cloned DNA probes from Shope fibroma virus (SFV) and myxoma virus. Under high stringency conditions for Southern blotting such that SFV probes do not cross-hybridize with myxoma virus DNA, it is demonstrated that greater than 90% of the MRV genome has been derived from myxoma virus, and that approximately 10 kb of SFV-derived sequences have substituted for a similar amount of myxoma sequences. Mapping of the MRV genome indicates that the SFV sequences are present in two regions of the genome, one in each copy of the MRV terminal inverted repeat sequence. Furthermore, fine mapping studies of the integration sites for SFV into the myxoma background show that these SFV sequences are not symmetrical with respect to the left and right genomic termini. At the left end, 4 kb of SFV-derived DNA maps between 6 and 10 kb from the terminus, while at the right end about 5.5 kb of SFV sequences are found to extend at least 1 kb further toward the unique internal sequences. Based on this asymmetrical bipartite distribution of SFV sequences in MRV, a two-stage model to rationalize the origin of MRV is proposed. This model postulates an initial recombination event similar to gene conversion between myxoma and SFV at the right terminus of myxoma, followed by an incomplete transposition of only part of these SFV sequences to the left terminus. PMID- 2981445 TI - A comparative analysis of radiation- and virus-induced leukemias in BALB/c mice. AB - Endogenous murine leukemia virus (MuLV) proviral copies were analyzed in thymomas induced in normal BALB/c (Fv-1b) and in Fv-1n congenic mice by X-irradiation. Both strains of mice developed leukemia with similar kinetics, indicating that N tropism of endogenous MuLV was not a rate-limiting factor in development of disease. Southern blot analysis, using a probe specific for ecotropic virus and for ecotropic-specific sequences retained in pathogenic, env-recombinant viruses, showed that the majority of radiation leukemias lacked newly acquired, clonally integrated, proviruses. This was in contrast to virus-induced leukemias, which routinely exhibited several new proviral integration sites. When an internal proviral DNA restriction fragment was monitored, some radiation leukemias showed evidence of nonclonal infection, accounting for more frequent isolation of infectious virus from such leukemias. Differences in expression of T-cell surface antigens were found in X-ray-induced and virus-induced leukemias. All radiation leukemias were TL positive, whereas virus-induced leukemias were primarily negative for TL. Some differences were also found in Lyt-1 and Lyt-2 expression. The data as a whole suggest that, in the majority of cases, radiation leukemogenesis is not initiated by a viral route--that is, the sort of viral mechanism for which exogenous infection by known pathogenic MuLV is the paradigm. PMID- 2981447 TI - Antigenic conservation and divergence between the viral-specific proteins of poliovirus type 1 and various picornaviruses. AB - Immuneprecipitation analyses of various picornavirus-infected cell lysates were performed using antisera to poliovirus type 1-specific structural and nonstructural proteins. The results showed differing patterns of antigenic conservation and divergence. However, the VP3 and 2C polypeptides were strongly antigenically conserved among the large majority of these viruses. This conservation was especially notable given the degree of divergence exhibited by the other viral proteins and may be due to environmental pressure exerted by interaction with the host cell. The results, furthermore, allowed for an analysis of the evolutionary relationship of the tested viruses. This analysis showed a particularly strong antigenic relationship between the proteins of the poliovirus group and coxsackievirus A21 as well as a weaker, but significant, relationship with coxsackieviruses B1 and B3. PMID- 2981448 TI - Lytic viruses infecting a Chlorella-like alga. AB - A number of viruses which form plaques on the unicellular, eukaryotic, Chlorella like green alga, strain NC64A, were isolated from fresh water ponds and rivers in Illinois, North Carolina, and South Carolina. The viruses were large polyhedrons (160 to 190 nm in diameter) and contained dsDNA genomes of ca. 300 kbp. All of the viral DNAs hybridized with DNA from the previously described PBCV-1 virus. However, the viruses, even many of those isolated from the same water sample, could be distinguished from one another by DNA restriction endonuclease digestion. The viruses, including PBCV-1, were grouped into five classes by their resistance to certain DNA restriction endonucleases. Presumably the DNAs in the five classes contain different types or amounts of modified bases. PMID- 2981449 TI - Pseudotyping of dual-tropic recombinant viruses generated by infection of mice with different ecotropic murine leukemia viruses. AB - Using a new focal immunofluorescence assay (FIA) with monoclonal antibodies, dual tropic recombinant mink cell focus-inducing (MCF) retroviruses were specifically detected directly in mouse cells. With the FIA, MCF and ecotropic murine leukemia virus (MuLV) infectious centers were quantitated in noninoculated AKR/J mice and in mice inoculated with either ecotropic Friend- or Moloney-MuLV. Comparison of the titers obtained in mouse and mink cells suggested that pseudotyping of MCF viruses with ecotropic MuLV envelopes occurred at different levels in these three mouse models. Adult and newborn IRW mice inoculated with Friend-MuLV produced MCF viruses which were mostly pseudotyped with ecotropic MuLV envelopes. Newborn IRW mice inoculated with Moloney-MuLV produced MCF viruses in both spleen and thymus. Most Moloney-MCF viruses from spleens were pseudotyped with ecotropic MuLV envelopes, whereas thymic Moloney-MCF viruses were not. Noninoculated AKR/J mice spontaneously produced MCF virus at least 2 months before the onset of organ enlargement, and the majority of MCF viruses in both spleens and thymuses of these mice did not appear to be pseudotyped. PMID- 2981450 TI - Characterization of west nile virus persistent infections in genetically resistant and susceptible mouse cells. II. Generation of temperature-sensitive mutants. AB - Long-term persistent infections were established with the flavivirus, West Nile virus (WNV), strain E101, in embryofibroblast cultures derived from susceptible C3H/HE and congenic-resistant C3H/RV mice. Cultures were initially maintained by weekly subculture at 37 degrees, but at passage 6 sister cultures were shifted to 32 degrees. Virus progeny titers were observed to increase after the shift to 32 degrees indicating the possible presence of temperature-sensitive mutants. Temperature-sensitive mutants were found to arise in cultures of both susceptible and resistant cells. However, only in the resistant cultures did temperature sensitive virus become the majority population. Temperature-sensitive mutants did not appear to be essential for either initiation or maintenance of WNV-persistant infections. The resistant cells appear to provide an environment which is advantageous for the amplification of temperature-sensitive mutants. PMID- 2981451 TI - The involvement of a type-B retrovirus in the induction of thymic lymphomas. AB - A highly leukemogenic virus (DMBA-LV) (in vivo leukemogenic titer 1-5 X 10(6) IU/ml, and 35-40 days to thymic lymphoma detection) is produced by a chemical carcinogen-induced transplanted thymic lymphoma. The virus preparation is a mixture of a type-B retrovirus highly related to exogenous type-B retroviral isolates and a biologically defective type-C retrovirus. The DNA of DMBA-LV induced-tumors contains new type-B proviruses but no additional type-C proviruses could be detected. The leukemogenicity of DMBA-LV was completely neutralized by a monoclonal antibody against MMTV envelope glycoprotein, but was not affected by a broadly reacting Friend MuLV anti-gp70 serum which effectively neutralizes type-C ecotropic, xenotropic, and recombinant retroviruses and which completely abolishes the leukemogenic activity of Moloney leukemia virus. Three type-B mammary tumor-inducing retroviral isolates, while containing type-C retroviral sequences, were not leukemogenic. A further characterization of the type-C retroviral sequences present in DMBA-LV indicated that sequences characteristic of endogenous, nonxenotropic proviruses are present. In addition, using a variety of type-C-specific retroviral DNA probes, no evidence was obtained for the presence of a type-B-C-recombinant genome in DMBA-LV. Leukemogenesis was absolutely dependent upon the presence of a functional type-B retroviral envelope gp 52 and DMBA-LV does not appear to contain a leukemogenic retroviral type-C genome. PMID- 2981452 TI - The four C-terminal amino acids of the v-erbA polypeptide are encoded by an intronic sequence of the v-erbB oncogene. AB - The genome of avian erythroblastosis virus (AEV), a defective acute leukemia retrovirus, carries two distinct cell-derived oncogenes in the structure 5' delta gag-erbA-erbB-delta env3'. The nucleotide sequence of the v-erbA gene was recently reported. In order to determine the boundary between the two adjacent oncogenes, the sequence of the v-erbA/v-erbB junction of AEV was compared to that of a recombinant lambda phage containing a chicken cellular sequence representing the 5' part of c-erbB. The four C-terminal amino acids of v-erbA are in fact encoded by a c-erbB intron-derived sequence thus demonstrating that the virus acquired a truncated c-erbA gene. Furthermore the 7 to 10 amino acid residues upstream from the 4 C-terminal amino acids mentioned above appeared to be derived from env-related sequences. The splice acceptor site at the beginning of the only open reading frame for v-erbB is also present and functional in c-erbB when expressed to generate a truncated EGF (epidermal growth factor) receptor. Thus AEV joins a truncated erbA gene to a truncated erbB gene through env-derived sequences and intronic sequences from c-erbB. PMID- 2981453 TI - Indomethacin inhibits viral RNA and protein synthesis in cells infected with vesicular stomatitis virus. AB - Treatment of HEp-2 cells with 10(-3) M indomethacin prior to infection with vesicular stomatitis virus (VSV) blocked synthesis of viral RNA species and proteins. Host cell macromolecular synthesis was also inhibited by this antagonist of prostaglandin metabolism. The reversibility of this drug-mediated inhibition was a time-dependent process that could be prevented after removal of indomethacin if cells were exposed to low concentrations of cycloheximide which do not normally interfere with VSV replication. That indomethacin exerts its inhibitory effect at an early stage of infection is suggested by the finding that this drug can compromise viral transcription either in vitro or in vivo. PMID- 2981454 TI - Physicochemical properties of the DNAs of the fastidious adenovirus species 40 and 41. AB - The DNAs of the two fastidious adenovirus species 40 and 41 (Ad40 and Ad41) have been studied by restriction endonuclease cleavage, CsCl density gradient centrifugation, and liquid-phase hybridization. For Ad40 DNA, cleavage maps were constructed for the restriction endonucleases EcoRI, SalI, ClaI, BstEII, BclII, and PvuI. The EcoRI, SalI, ClaI, and XhoI maps could be established for Ad41 DNA. The buoyant density in CsCl of DNAs of the fastidious species is 1.711 g/cm3 corresponding to a G + C content of 52%. The homology of the DNAs from Ad40 and Ad41 proved to be 62-69%. These results are consistent with previous reports on antigenic and DNA restriction enzyme analysis in which the classification into two species was proposed (J. C. De Jong, R. Wigand, A. H. Kidd, G. Wadell, J. G. Kapsenberg, C. J. Muzerie, A. G. Wermenbol, and R.-G. Fritzlaff, J. Med. Virol. 11, 215-231 (1983), and I. Uhnoo, G. Wadell, L. Svensson, and M. Johansson, Dev. Biol. Stand. 53, 311-318 (1983]. PMID- 2981455 TI - Combined macroscopic and microscopic detection of viral genes in tissues. AB - A hybridization technique has been devised for detecting and quantitating viral genes in tissues that combines macroscopic and microscopic analyses in the same section. The method is based on dual labeling virus-specific probes with 125I and 35S to generate signals that can be detected both with X-ray films and nuclear track emulsions. The regions of increased hybridization evident in the X-ray film serve as a guide to the portion of the section that warrants microscopic examination. Detection of viral RNA in tissues with visna virus and viral DNA with hepatitis B virus are illustrated, and potential applications of this technique in virology and other disciplines are discussed. PMID- 2981456 TI - DNA sequence of the early E3 transcription unit of adenovirus 5. AB - The DNA sequence of the early E3 transcription unit of adenovirus 5 (Ad5) has been determined and it has been compared to Ad2, as published previously [J. Herisse, G. Courtois, and F. Galibert (1980), Nucl. Acids Res. 8, 2173-2192; J. Herisse and F. Galibert (1981), Nucl. Acids Res. 9, 1229-1240]. The E3 regions of Ad5 and Ad2 are quite homologous despite being nonessential for Ad growth in cultured cells. The major differences are "gaps" that exist either in Ad5 or Ad2 in intergenic regions. The conservation of sequences suggests that E3 plays a beneficial role in natural infection of humans. E3 appears to encode about seven to nine proteins; based on sequence, seven of these may be membrane proteins. Thus, E3 may be a transcription unit devoted to the synthesis of membrane proteins. The E3 genes lie essentially one after the other along the genome, and which gene is expressed from a given primary transcript is determined by the choice of the 3' end site and the 5' and 3' splice sites. Almost all E3 mRNAs contain nonfunctional AUGs that are 5' to the initiation codon. Codon usage is nonrandom. Although the CG dinucleotide frequency is low, CG clusters exist in the promoter and other regions. PMID- 2981457 TI - Analysis of the terminal sequences of the genome segments of four orbiviruses. AB - The dsRNA genome segments of bluetongue virus (BTV) types 1 and 20 and Ibaraki virus (a member of the epizootic haemorrhagic disease (EHD) serogroup) have conserved sequences of six bases at both of their 3' termini. One strand of all the genome segments analysed ends in 3'CAUUCA ... 5' while the other strand ends in 3'CAAUUU ... 5'. These conserved sequences are identical to those previously reported for BTV types 10 and 11 (A. Kiuchi, C. D. Rao, and P. Roy (1983), "Double-Stranded RNA Viruses" (R. W. Compans and D. H. L. Bishop, eds.), pp. 55 64. Elsevier, New York; C. D. Rao, A. Kiuchi, and P. Roy (1983), J. Virol. 46, 378-383). The 3' terminal sequences of segments 3 and 10 of the BTV type 1 genome were confirmed by the detection of exactly complementary sequences at the 5' termini of the ssRNA strands of opposite polarity. This also confirmed for these dsRNA segments (and by analogy for all the genome segments of these viruses) that the dsRNA molecules are fully base paired end to end. Using in vitro synthesised mRNA of BTV type 1 in annealing experiments with the two ssRNAs separated from each of the individual genome segments, it was shown that in each case the strand ending in 3'CAUUCA ... 5' is of the same polarity as the mRNA (+ve), while the strand ending in 3'CAAUUU ... 5' is of the opposite (-ve) polarity. The fourth virus analysed (Tilligerry virus, a member of the Eubenangee serogroup) only had five conserved bases at the 3' termini of one strand of its genome segments (3'CAU-CA ... 5') and three conserved bases at the 3' termini of the other strand (3'CA--U ... 5'). Considerable sequence homology was found in the near-terminal nonconserved regions of comparable genome segments from the different viruses, particularly between the different BTV types. There was little evidence, however, for absolute conservation of "segment specific" sequences in these regions of the RNA. PMID- 2981458 TI - Value of heparinized fine-needle aspiration biopsy in liver malignancy. AB - The accuracy of diagnosis of hepatic malignancy by percutaneous aspiration biopsy was compared using heparinized and nonheparinized 22-gauge needles. When a heparinized instrument was used, it was able to accurately diagnose malignancy and cytologic types of exfoliative cells. Heparinization also made it possible to recover small tissue fragments and make ultrathin sections for histologic examination. In this way, material suitable for cytologic and histologic examination was obtained from 100% and 95%, respectively, of 59 patients suspected of having hepatic malignancy. A correct diagnosis was made cytologically in 92.5% of patients with hepatic malignancy and in 89.5% of those from whom histologic material was obtained. From a combination of histologic and cytologic results, the overall diagnostic rate for hepatic malignancy was increased to 95%. This procedure proved to be a reliable method for diagnosis of hepatic malignancy. PMID- 2981459 TI - Left ventricular volume determination from single-photon emission computed tomography. AB - To compare the accuracy of single-photon emission computed tomography (SPECT) with that of contrast cineangiography in measuring left ventricular end-diastolic volume, 25 consecutive patients undergoing catheterization for coronary artery or valvular heart disease were first evaluated scintigraphically. A simple, count corrected voxel counting technique was used to derive left ventricular end diastolic volume from transaxial SPECT slices. SPECT volume values showed a high degree of correlation with those determined by angiography (r = 0.969), with a standard error of the estimate of 23 ml. SPECT offers a highly accurate and essentially noninvasive method for measuring chamber volumes that is independent of geometric assumptions about ventricular configuration and chest wall attenuation and does not require blood sample counting. PMID- 2981460 TI - Thoracic duct opacification for CT scanning. PMID- 2981461 TI - Preservation of ischemic myocardium by a new converting enzyme inhibitor, enalaprilic acid, in acute myocardial infarction. AB - Enalaprilic acid (MK-422), the biologically active diacid of the converting enzyme inhibitor enalapril, was studied in myocardial ischemia (MI). Acute left coronary artery ligation was produced in 62 male Sprague-Dawley rats, and infarct size was determined by left ventricular free wall (LVFW) creatine kinase (CK) activity. Administration of enalaprilic acid (2 mg/kg) 2 minutes and 24 hours after MI significantly blunted the reduction in LVFW CK activity at 48 hours after ligation, when compared to the MI rats given vehicle (6.4 +/- 0.5 vs 4.7 +/ 0.2 IU/mg protein, respectively; p less than 0.01). The percentage of LVFW spared was significantly (p less than 0.01) increased from 28 +/- 2% to 45 +/- 5% by MK-422. MK-422 also significantly blunted the loss of LVFW CK activity 48 hours after a coronary ligation (10 minutes) followed by reperfusion, when compared to vehicle (10.1 +/- 0.6 vs 8.3 +/- 0.6 IU/mg protein, respectively; p less than 0.05). This represents a significant increase in the percentage of LVFW spared, 65 +/- 5% vs 85 +/- 6% (p less than 0.05). These data indicate a significant protective action afforded by MK-422 in two different protocols of ischemic damage to the myocardium and suggest a role for the renin-angiotensin system in the extension of ischemic damage. PMID- 2981462 TI - Immediate converting-enzyme inhibition with intravenous enalapril in chronic congestive heart failure. AB - To test the hypothesis that intravenous enalapril is a useful pharmacologic probe of the renin angiotensin system, intravenous enalapril was administered to 9 patients with severe congestive heart failure (CHF). This produced abrupt and complete blockade of converting enzyme, with peak effect occurring at 30 minutes, as reflected by increases of plasma renin activity (from 16.8 +/- 6 to 86.6 +/- 23 ng/ml/hour) and decreases of plasma aldosterone levels (from 46 +/- 14 to 25 +/- 6 ng%) (both p less than 0.05). With reduction of angiotensin II--mediated vasoconstriction, systemic vascular resistance decreased markedly (from 1,974 +/- 233 to 1,400 +/- 136 dyne s cm-5) and cardiac index was improved (from 1.88 +/- 0.9 to 2.20 +/- 0.21 liters/min/m2) (both p less than 0.05). The time course of angiotensin II levels suggested that the lack of a cumulative effect from additive doses of intravenous enalapril was a reflection of complete inhibition of converting enzyme. One patient did not respond to enalapril; despite comparable hemodynamic severity of CHF, the renin-angiotensin system was not activated in this patient. Thus, intravenous enalapril is capable of rapid and complete inhibition of converting enzyme for the accurate assessment of angiotensin II--mediated vasoconstriction in patients with severe CHF. PMID- 2981463 TI - Role of vasodilators in the treatment of congestive heart failure. AB - Appreciation of the important role played by peripheral vasoconstriction in the pathophysiology of congestive heart failure (CHF) has led to the widespread use of vasodilators as treatment. Short-term studies show that, regardless of the vasodilator used, the arterial and venous dilatation produced invariably results in improvement in the hemodynamic status of patients. This short-term response, however, does not automatically translate to long-term clinical improvement. The reasons for this are not well understood but such factors as differing mechanisms of action, development of tolerance and unique patterns of regional redistribution of blood flow may all play a modifying role in differentiating one vasodilator from another. Nevertheless, a number of controlled trials have demonstrated sustained symptomatic and functional improvement when vasodilators such as the converting enzyme inhibitors or nitrates are given to patients with CHF. PMID- 2981464 TI - Clinical and hemodynamic experience with enalapril in congestive heart failure. AB - The renin-angiotension system is activated in many patients with congestive heart failure (CHF), resulting in angiotensin-mediated vasoconstriction and aldosterone mediated sodium and water retention. To evaluate the effectiveness of enalapril, a new converting enzyme inhibitor, enalapril was administered to patients either orally or intravenously in a single dose, and hemodynamic and hormonal responses were measured. Patients were then placed on oral enalapril therapy for 1 month, and treadmill exercise duration and invasive hemodynamics were compared with baseline pretreatment data. With single-dose administration, both oral and intravenous enalapril reduced systemic vascular resistance and increased cardiac output. However, the effects of oral enalapril were not manifest for 3 to 4 hours, because oral enalapril is a pro-drug form that requires hepatic deesterification. In contrast, intravenous enalapril resulted in significant hemodynamic and hormonal changes 15 to 30 minutes after administration. During long-term therapy, enalapril was associated with improved symptomatology, increase of treadmill exercise duration and sustained hemodynamic improvement. Enalapril was effective therapy for chronic CHF. Optimal short-term response may require coadministration of both intravenous and oral preparations of enalapril; however, the magnitude of the short-term response was comparable for both preparations. Long-term therapy is most effective when the drug is administered as a twice-daily regimen. PMID- 2981465 TI - New positive inotropic drugs for the treatment of congestive heart failure. AB - Positive inotropic agents can stimulate the severely depressed myocardium in late stages of heart failure. However, symptomatic benefits are only gained by improvement in the deranged peripheral circulation, which produces symptoms and limitations. In augmenting cardiac output and reducing filling pressures, the effects of positive inotropic agents and vasodilators are similar and additive, and the "contractile reserve of the heart" in response to inotropic stimulation may limit efficacy of these agents. Although symptomatic benefits occur in patients with severe heart failure after improvement in peripheral blood flow distribution, survival may not be altered, because this appears to be determined more by the amount of myocardial damage and its progression, and neither of these is affected by either inotropic agents or vasodilators. Indeed, in early stages of heart failure, therapy must be redirected toward preventing further myocardial cell loss rather than stimulating pump function. PMID- 2981466 TI - Nonendemic adult T-cell leukemia/lymphoma in the United States: report of two cases and review of the literature. AB - Adult T-cell leukemia/lymphoma (ATLL) is a recently described distinct clinicopathologic entity characterized by a leukemic or lymphomatous proliferation of hyperlobulated peripheral T-cells, which is usually widespread at presentation and is associated with infection by a type C retrovirus. ATLL rarely is described outside of endemic regions, which include southwestern Japan, the Caribbean region, and the southeastern United States. The authors report the clinical, pathologic, and immunologic features of two cases of nonendemic ATLL that occurred in patients from the midwest United States. One patient was a 16 year-old white girl from rural Iowa, and the other was a 46-year-old white man from rural Minnesota. The features of 13 other probable nonendemic ATLL cases from the United States were compiled and reviewed. In the United States, nonendemic ATLL occurred in widespread geographic locations, affected mostly white people, and was characterized by an aggressive course with generalized adenopathy, blood and bone marrow involvement, and hepatosplenomegaly at presentation. Skin involvement was present in one-fourth of the patients. Hypercalcemia was rare. Although antibodies to type C retrovirus were detected in three of the five patients tested, the available data is not sufficient to establish a conclusive association between nonendemic ATLL and type C retrovirus infection. PMID- 2981467 TI - Reproducibility of the French-American-British classification of acute leukemia: the Southwest Oncology Group Experience. AB - After initial French-American-British (FAB) diagnosis by a multiinstitutional Southwest Oncology Group panel, slides of acute leukemia cases were recirculated to panel members for second review. The reproducibility of the FAB classification is analyzed. The classification is reproducible in the 70% range in panel reviewer hands and allows remarkable reproducibility in the morphologic and cytochemical distinction of acute lymphoid leukemia (ALL) from acute myeloid leukemia (AML). The limitations of a morphologic and cytochemical classification of acute leukemia are discussed. A simplification of the FAB system, merging M1, M2, and M4 as M7, is proposed; this simplification improves the system's reproducibility. PMID- 2981468 TI - Lymphoma of host origin in a marrow transplant recipient in remission of acute myeloid leukemia and receiving cyclosporin A. AB - Lymphomas are an uncommon complication of solid organ transplantation and rarely occur after marrow transplantation. When post-marrow transplant lymphomas have occurred, they have been of donor cell origin and when sought, Epstein-Barr virus DNA has been found in the tumor. A 21-year-old woman developed a poorly differentiated lymphocytic lymphoma 6 months after bone marrow transplantation for acute myeloid leukemia in remission. Cyclosporin A had been used as an immunosuppressant. A chromosomal polymorphism demonstrated that the tumor was of host origin and contained a monoclonal tumor marker, 46,XX INV 4 (p16q12). The tumor did not contain the DNA of the Epstein-Barr virus. PMID- 2981469 TI - Small noncleaved B cell Burkitt-like lymphoma with chromosome t(8;14) translocation and Epstein-Barr virus nuclear-associated antigen in a homosexual man with acquired immune deficiency syndrome. AB - This case report describes new manifestations of the acquired immune deficiency syndrome (AIDS) in a promiscuous homosexual man. Investigation of upper gastrointestinal bleeding in the patient lead to discovery of a high-grade, small, noncleaved cell (Burkitt-like) gastroduodenal lymphoma with visceral and extralymphatic extension. Specific phenotyping of the lymphoma revealed that it was a monoclonal B cell lymphoma of mu kappa isotype. An in vitro cell line was established that was Epstein-Barr virus nuclear-associated antigen-positive. The lymphoma cells displayed a t(8;14) translocation similar to endemic African Burkitt lymphoma. Epstein-Barr virus genomes were identified in the lymphoma and an axillary lymph node biopsy specimen by molecular hybridization. These data strongly suggest that Epstein-Barr virus actively infected this patient. However, he showed normal Epstein-Barr virus-specific serologic responses, indicating an immune defect against the virus. PMID- 2981470 TI - Localization of transferrin receptor in the chorionic villi of complete molar pregnancy. AB - Transferrin receptors have been identified on the villous trophoblast of normal chorionic villi by immunohistochemical techniques. The current study investigated the expression of transferrin receptors on molar chorionic villi by using a monoclonal antibody in immunofluorescence assays. The villous trophoblast of molar chorionic villi was brightly positive for transferrin receptor in the immunofluorescence assay. The presence of transferrin receptors on molar villous trophoblast may influence the immunologic relationship between molar and host tissues. PMID- 2981471 TI - Functional defects in phagocytic cells following thermal injury. Application of flow cytometric analysis. AB - Defective phagocytic cell function may partially account for the morbidity and mortality associated with thermal injury. In experimental thermal injury in the rat, small circulating blood volumes increase the difficulty in obtaining significant data. Furthermore, purification and or elicitation procedures have the potential for altering the cell surface characteristics and/or the functional response of the cell in question. We have examined the circulating neutrophils and pulmonary alveolar macrophages of anesthetized rats following a 16-20% body surface area scald injury to the shaved back. The circulating neutrophils of thermally injured rats were examined by flow cytometry following stimulation with phorbol myristate acetate (PMA) (100 ng/ml) in terms of the change in fluorescence intensity of the potentiometric cyanine dye, dipentyloxocarboxyanine and the formation of the oxidized product of 2',7'-dichlorofluorescin diacetate loaded cells. The alveolar macrophages were examined after stimulation with PMA (100 ng/ml) in terms of the change in fluorescence intensity of the potentiometric dye, dipropylthiodicarbocyanine and the generation of superoxide production, as assessed by the superoxide dismutase inhibitable reduction of cytochrome c. Both cells exhibited a profound inhibition of cell function 4 hours after the insult, with partial return toward control values at later time points. Furthermore, the plasma of thermally injured rats, 4 hours after the burn was inhibitory to normal rat neutrophils. Fluorescent compounds suggestive of in vivo lipid peroxidation were maximally detectable at this time point. Further research is needed to establish the role of these products in the induction of phagocytic cell dysfunction. PMID- 2981472 TI - Interaction between enteroviruses and human endothelial cells in vitro. Alterations in the physical properties of endothelial cell plasma membrane and adhesion of human granulocytes. AB - Fluorescent molecular probes were used for study of the interaction between enteroviruses (Echo 9, Echo 12, and Coxsackie B3 virus) and human endothelial cells in monolayer culture. With the use of the monomer-excimer method with pyrene decanoic acid it was shown that a marked dose-dependent restructuring of the plasma membrane occurred following addition of virus to the endothelial cells. This took the form of an increase in the lipid surface available to the lipophil reporter molecules, probably due to an alteration in the domain structure of the plasma membrane caused by insertion of virus capsid proteins. Experiments with diphenylhexatriene indicated that the enteroviruses had only a slight tendency to make the plasma membrane of the endothelial cell more fluid. Concomitant with these alterations in the biophysical properties of the membrane, a virus-induced increase in granulocyte adherence to the endothelial cells was observed for all three enteroviruses studied. Possible mechanisms for this elevated adherence are discussed, as well as the significance of the results for the phenomenon of virus-induced granulocytopenia. PMID- 2981475 TI - Lack of fructose-1,6-bisphosphatase activity in LLC-PK1 cells. PMID- 2981473 TI - Extracellular matrix and epithelial differentiation of Wilms' tumor. AB - Different histologic types of 82 Wilms' tumors were graded on the basis of the histologic pattern. Representative tumors of each group were analyzed by the immunoperoxidase method for evaluation of the histogenesis of Wilms' tumor and the value of antibodies against extracellular matrix (ECM) components (collagens I and III, laminin, fibronectin) in differential diagnosis of different types of Wilms' tumors. The tubules of classic Wilms' tumor expressed laminin, which could be seen also in and around some blastemal cells. Blastema and tubules were negative for interstitial collagens, but Type I and III collagen were prominent in the fibrovascular stroma. The monomorphous tubular, psammomatous and rosetting tumors expressed laminin, but no interstitial collagens. In sarcomas, only the blastemal variant of spindle-cell sarcomas was negative for interstitial collagens, which were abundantly seen in all other sarcomas. While spindle-cell sarcomas were devoid of laminin, the highly malignant rhabdoid and clear-cell sarcomas expressed laminin in a characteristic dotted fashion. Staining for fibronectin gave varying results and had therefore only a limited value in distinguishing different types of Wilms' tumors. However, the antibodies against interstitial collagens and against the basement membrane glycoprotein laminin turned out to be a useful adjunct in differential diagnosis and classification, especially of sarcomatoid Wilms' tumors. The basement membrane of normal nephrons is similar to that in tubules of triphasic Wilms' tumor, but the ECM of blastemas is different. This transformed phenotype might represent a maturation arrest of the blastemal cell when compared with the expression of proteins during normal nephrogenesis. PMID- 2981474 TI - Receptor occupancy vs. induction of Na+-K+-ATPase and Na+ transport by aldosterone. AB - In the urinary bladder of the toad Bufo marinus aldosterone (between 0.8 and 100 nM) stimulates Na+ transport [half-maximal induction concentration (K1/2) = 6.5 nM]. At low hormone concentrations (0.8-8 nM), the increase of Na+ transport between 0.75 and 2.5 h is accompanied by a fall in transepithelial resistance (R). Higher hormone concentrations (30-800 nM) induce an additional resistance independent fraction of Na+ transport within 2.5-8 h. From 6 h on, aldosterone (between 0.2 and 20 nM) stimulates in the same tissue the biosynthesis rate of the alpha- and beta-subunits of Na+-K+-ATPase (K1/2 = 3 and 1.5 nM, respectively). New pump synthesis is thus not a prerequisite for the early mineralocorticoid response but might be linked to the late transport event. The mineralocorticoid response is usually ascribed to interaction with the higher affinity type 1 receptor. In the present study we show, however, that at least 55% of the overall Na+ transport response is linked to nuclear occupation of the lower affinity type 2 receptors [dissociation constant (Kd) = 50 nM, maximum number of binding sites (Nmax) = 315 fmol/mg protein]. Distinct aldosterone effects, such as the fall in R and the increase in Na+-K+-ATPase synthesis, are more closely related to occupation of type 1 receptors (Kd = 0.3 nM, Nmax = 23 fmol/mg protein). At maximal induction of these latter parameters, only about 20% of type 2 receptors are occupied. These results suggest that both types of aldosterone receptors are involved in the mediation of the full mineralocorticoid response: type 1 in the early and late and type 2 particularly in the late tissue response. PMID- 2981476 TI - Electrophysiological effects of propionate and bicarbonate on gallbladder epithelium. AB - The effects of propionate and HCO3- on cell membrane potentials in Necturus gallbladder epithelium were investigated using microelectrode techniques in vitro. Bilateral exposure to either propionate or HCO3- (25 mM each) hyperpolarized both cell membranes by about 12 mV. Mucosal addition of either substance produced cyclic changes in voltage of both cell membranes, which attenuated spontaneously, whereas serosal addition caused sustained hyperpolarization. By intraepithelial cable analysis it was shown that both cell membrane conductances rose during the hyperpolarization. Experiments using substitution of mucosal K+ for Na+ revealed that the relative K+ permeability (PK) of the apical membrane was enhanced during the hyperpolarization induced by mucosal (or serosal) propionate (or HCO3-). These effects are mediated by increases in PK at both membranes, with a larger basolateral effect. We suggest that this mechanism accounts for the higher cell membrane potential values measured in epithelia bathed in HCO3--or propionate-containing solutions. Inasmuch as both propionate and HCO3- stimulate fluid absorption in gallbladder epithelium, the increase in cell membrane PK may represent an adaptive response of the cells to regulate their solute content. PMID- 2981477 TI - Specific binding sites for ovine prolactin in three amphibian cell lines. AB - Established cell lines (TB-6c and TB-M) obtained by continuous culture of epithelial cells from toad Bufo marinus urinary bladder, which, in culture, maintained a high degree of functional differentiation, exhibited a significant number of high-affinity (KA = 1-2 X 10(10) M-1) binding sites detected both with radioiodinated (125I) ovine prolactin (oPRL) and human growth hormone (hGH). Binding capacity was higher in the case of TB-6c cells (7,573 +/- 581 sites/cell) than with the TB-M cells (1,160 +/- 87). Similarly, binding sites for oPRL were characterized on Xenopus laevis kidney-derived cell line A6. With oPRL used both as tracer and standard, significant cross-reaction was observed with hGH, less with human or rat prolactin (PRL), and none with human chorionic somatomammotropin, bovine growth hormone, and rat luteinizing hormone or follicle stimulating hormones. B. marinus pituitary extracts completely displaced the binding of 125I-oPRL to toad bladder binding sites. This finding of specific sites for PRL on amphibian bladder and kidney cells confirms that PRL exerts specific biological actions for the control of electrolyte and water metabolism in the amphibians. PMID- 2981478 TI - Coenzyme A metabolism. AB - The metabolism of coenzyme A and control of its synthesis are reviewed. Pantothenate kinase is an important rate-controlling enzyme in the synthetic pathway of all tissues studied and appears to catalyze the flux-generating reaction of the pathway in cardiac muscle. This enzyme is strongly inhibited by coenzyme A and all of its acyl esters. The cytosolic concentrations of coenzyme A and acetyl coenzyme A in both liver and heart are high enough to totally inhibit pantothenate kinase under all conditions. Free carnitine, but not acetyl carnitine, deinhibits the coenzyme A-inhibited enzyme. Carnitine alone does not increase enzyme activity. Thus changes in the acetyl carnitine-to-carnitine ratio that occur with nutritional states provides a mechanism for regulation of coenzyme A synthetic rates. Changes in the rate of coenzyme A synthesis in liver and heart occurs with fasting, refeeding, and diabetes and in heart muscle with hypertrophy. The pathway and regulation of coenzyme A degradation are not understood. PMID- 2981479 TI - Reduction of pituitary GnRH receptors in immature rats treated with monosodium glutamate. AB - The number of pituitary gonadotropin-releasing hormone (GnRH) receptors increases during sexual maturation in the rat and probably reflects changes in hypothalamic GnRH secretion. As GnRH is synthesized in various hypothalamic nuclei, including the arcuate nucleas (ARC), we investigated the effects of monosodium glutamate (MSG)-induced lesions of the ARC in the rat. In males and females treated with MSG during the first 10 days of life, GnRH receptor content (GnRH-RC) was unchanged from controls at 10 days but was decreased at 20 and 30 days of age (P less than 0.01). Serum concentrations of luteinizing hormone (LH) were similar in MSG-treated and control males but were significantly lower in 10-day-old females (P less than 0.01). Injections of GnRH (3 micrograms every 8 h on days 18 and 19) restored GnRH-RC to control values in MSG-treated rats. Both MSG and untreated control rats showed similar LH responses to acute injections of GnRH, but responses were attenuated (P less than 0.05) after 2 days pretreatment with GnRH in rats that had received MSG. Ovarian GnRH-RC was similar in both MSG-treated and untreated controls. These data indicate that MSG-induced lesions of the ARC reduce pituitary GnRH-RC in immature rats, and the more marked effects in females suggest a more significant role in the ARC in the control of GnRH secretion during maturation in females. The lack of MSG-induced changes in ovarian GnRH-RC indicates that GnRH from the arcuate nucleus is not responsible for the increase in ovarian GnRH receptors seen during sexual maturation. PMID- 2981480 TI - Effects of benzyl alcohol on PTH receptor-adenylate cyclase system of canine kidney. AB - In many systems, perturbations of membrane architecture by changes of lipid and phospholipid composition have been shown to alter the activity of membrane-bound enzymes. The present studies examined the effect of benzyl alcohol, an agent that has been shown to increase membrane fluidity, on the parathyroid hormone (PTH) sensitive adenylate cyclase system of canine kidney. Benzyl alcohol progressively increased basal adenylate cyclase activity up to fourfold and maximal enzyme activity in the presence of PTH, GTP, guanylimidodiphosphate, and sodium fluoride by four- to sixfold. In the presence of 20 mM Mn2+ (no Mg2+), conditions under which enzyme activity is devoid of influence of guanine nucleotides or hormones, benzyl alcohol was without effect. PTH binding was increased by 25% in the presence of benzyl alcohol without a change in binding affinity. Fluorescent polarization studies using diphenylhexatriene showed a decrease in fluorescence anisotropy in the presence of benzyl alcohol. The results suggest that benzyl alcohol facilitates the interaction of the components of the adenylate cyclase system, presumably by increasing membrane fluidity. Alterations of membrane fluidity may be a potent means of regulating hormone sensitive adenylate cyclase activity. PMID- 2981481 TI - Calcium, a "third messenger" of cAMP-stimulated adrenal steroid secretion. AB - This study examines the role of extracellular calcium and calcium mobilization from intracellular stores in mediating cAMP-stimulated steroid secretion by rat adrenal glomerulosa cells (GC) and fasciculata cells (FC). When GC were incubated acutely in a calcium-deficient buffer, cAMP failed to significantly increase aldosterone secretion above base line. Aldosterone secretion, however, rose from 17 +/- 2 to 32 +/- 4 ng/10(6) cells (P less than 0.01) as calcium in the medium was increased from 0 to 3.5 mM. In contrast, cAMP-stimulated corticosterone production by FC was not influenced by changes in the external calcium concentration. Lanthanum (10(-4) M), an inhibitor of calcium influx, reduced cAMP stimulated aldosterone secretion from 69 +/- 10 to 42 +/- 5 ng/10(6) cells (P less than 0.01) but failed to alter cAMP-stimulated fasciculata steroidogenesis. Depletion of intracellular calcium stores, achieved by incubating with EGTA, markedly blunted cAMP-stimulated corticosterone secretion in GC from 666 +/- 126 to 32 +/- 6 ng/10(6) cells (P less than 0.01), and cAMP-stimulated corticosterone secretion in FC from 2,223 +/- 407 to 414 +/- 58 ng/10(6) cells (P less than 0.01). TMB-8, a putative inhibitor of intracellular calcium mobilization, markedly inhibited (P less than 0.01) cAMP-stimulated aldosterone secretion by GC from 469 +/- 31 to 48 +/- 8 ng/10(6) cells and corticosterone secretion by FC from 9,867 +/- 1,821 to 2,832 +/- 586 ng/10(6) cells. These observations suggest that cAMP activation of adrenal steroidogenesis requires the release of calcium from intracellular stores.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981482 TI - Colonic potassium and chloride secretion: role of cAMP and calcium. AB - Stripped rabbit colonic mucosa was studied in vitro in Ussing chambers to further investigate the role of Ca in regulating K and Cl secretion stimulated by the divalent cation ionophore A23187, prostaglandin E1 (PGE1), or 8-bromo-cAMP (8BrcAMP). To assess the effects of these secretagogues on the paracellular shunt permeability, we measured the Na concentration dependence of the serosal-to mucosal Na flux in the absence or presence of these stimuli. Results from these studies reveal that changes in net K and Cl secretion produced by secretory stimuli cannot be accounted for by a change in shunt permeability. The possible involvement of Ca in the secretory response of the colon to these stimuli was investigated by measuring the changes in Cl and K transport elicited by A23187, PGE1, or 8BrcAMP in the absence or presence of trifluoperazine (10(-4) M) added to the serosal bathing solution. Trifluoperazine alone did not significantly alter basal Na or Cl fluxes or short-circuit current (Isc) but did decrease transepithelial conductance (Gt) and the serosal-to-mucosal K flux. Pretreatment of the tissues with trifluoperazine significantly reduced or abolished the changes in K fluxes elicited by A23187, 8BrcAMP, or PGE1 without altering the changes in Cl transport, Isc, and Gt. These results suggest that K secretion induced by these secretagogues involves an increase in intracellular Ca concentration and may be mediated by calmodulin. PMID- 2981483 TI - Colonic glycoprotein secretion and calmodulin-acceptor proteins in the reserpine treated rat. AB - The rate of radioactive precursor-labeled colonic glycoprotein secretion in chronically reserpine-treated versus saline-injected control rats was examined. Everted colonic sacs prepared from reserpine-treated rats were found to be hypersecretory, exhibiting a basal rate of glycoprotein secretion that was threefold higher than control everted sacs. Furthermore, glycoprotein secretion in control tissue was stimulated by the secretagogue carbachol, and this stimulation was precluded by 10 microM trifluoperazine, the calmodulin antagonist. Reserpine-treated tissue, in contrast, was refractory to treatment with carbachol as well as trifluoperazine. While reserpine-treated and control colonic mucosae were demonstrated to contain equivalent levels of calmodulin via radioimmunoassay, reserpine-treated tissue was determined to lack two calmodulin acceptor proteins with molecular weights of 29 and 47 kilodaltons. The data suggest that the mechanism by which reserpine elicits this cystic fibrosislike, hypersecretory state of glycoprotein secretion within the colonic mucosa entails the loss of calmodulin function in the regulation of this secretory process. We speculate that the biochemical defect present in cystic fibrosis could also entail such a loss of calmodulin function in the regulation of glycoprotein secretion. PMID- 2981484 TI - Patients' rights: a cultural challenge to Western psychiatry. AB - To some degree, all psychiatric encounters, by constraining behavioral variation and autonomous decision making, threaten a personhood already impaired by illness. The challenge presented by this and other factors in Western psychiatry is how to resolve the ethical dilemmas of justice, autonomy, and beneficence inherent in contemporary hospitalization and treatment practices for severely mentally ill persons. The author discusses this challenge in terms of the physician-patient contract, clinical standards for limiting autonomy, and the Kantian concept of equality of mutual respect. The legal code in relation to psychiatry has evolved into a contemporary interpreting mechanism for Kantian cultural tradition. PMID- 2981485 TI - Serologic survey of rotavirus, Norwalk agent and Prototheca wickerhamii in wastewater workers. AB - Analysis of paired sera from 48 wastewater workers and controls who reported gastrointestinal illness did not reveal any excess of seroconversions to Norwalk agent or to rotavirus. Inexperienced wastewater-exposed workers had higher levels of antibody to Norwalk agent than did experienced and control workers and those with high and medium aerosol exposure had higher titres than those in the low aerosol category. Analysis for Prototheca antibody titres was essentially negative. PMID- 2981486 TI - Fibrolamellar hepatoma. AB - We found the clinical features of fibrolamellar hepatoma similar to those of nonfibrolamellar hepatoma with the exception of patient age. Although the histopathologic findings of fibrolamellar hepatoma are distinct and easily recognizable, we found that fibrolamellar hepatomas may be histologically heterogeneous. The overall length of survival of patients with fibrolamellar hepatoma was greater than that of patients with nonfibrolamellar hepatoma, but the survival resection was similar, regardless of histologic characteristics. Differences in overall survival between histologic subtypes probably reflects differences in the rate of resectability between fibrolamellar and nonfibrolamellar hepatomas. PMID- 2981487 TI - Prolonged hyporesponsiveness of vascular smooth muscle contraction after halothane anesthesia in rabbits. AB - Halothane diminishes smooth muscle contractility in vascular tissue. In order to further characterize this phenomenon we undertook a series of in vivo and ex vivo experiments. Pressor dose-response curves to the selective alpha 1-adrenergic agonist, phenylephrine, were constructed in groups of rabbits before, during and 2 hr after halothane anesthesia and the dose of phenylephrine that induced a 25 torr increase in mean arterial pressure (ED25) was derived by polynomial regression analysis. ED25 torr increased significantly during halothane anesthesia, and rabbits remained in this insensitive state when the ED25 was assessed 2 hr after anesthesia. The halothane-induced loss of responsiveness was corroborated by ex vivo experiments utilizing aortic rings from halothane anesthetized rabbits. The maximal contraction to norepinephrine (NE) was significantly lower in halothane-treated aortic rings and only slowly returned to normal by 4 hr. The EC50 (the dose causing a 50% maximal contraction) for NE was significantly greater in aortic rings from halothane-anesthetized rabbits. This loss of sensitivity, reflected by the higher EC50 was not restored by 4 hr of ex vivo incubation in a halothane-free medium. We conclude that halothane induces loss of sensitivity to adrenergic agonists that persists for several hours after termination of the halothane anesthetic. PMID- 2981488 TI - Enflurane and adenosine 3',5'-cyclic monophosphate metabolism in pancreatic islets. AB - The effects of enflurane at a concentration of 4% by volume in the gas phase upon rates of insulin secretion were correlated with islet contents of adenosine 3',5' cyclic monophosphate (cyclic AMP). At this concentration, enflurane was without effect upon the basal rate of insulin release in the presence of 2 mM glucose. By contrast, the anesthetic led to 84 and 86% inhibitions, respectively, of rates of insulin secretion stimulated by 20 mM glucose and 20 mM glucose plus 1 mM theophylline. Enflurane also led to small reductions in islet cyclic AMP content under all conditions of incubation, and a statistically significant (P less than 0.02) effect of the anesthetic in lowering cyclic AMP content was seen in islets exposed to 20 mM glucose plus 1 mM theophylline. Enflurane was without effect upon the activity of the low Km cyclic AMP phosphodiesterase of the islet under the conditions employed. By contrast, the anesthetic led to significant inhibitions of both basal and fluoride-stimulated islet adenylate cyclase activity (34%, P less than 0.01 and 23%, P less than 0.005, respectively). It is concluded that under the conditions employed, enflurane leads to inhibition of islet adenylate cyclase activity and to small resultant reductions in islet cyclic AMP content. These effects appear to be of insufficient magnitude to explain completely the observed degree of insulin secretory inhibition by the anesthetic. PMID- 2981489 TI - Anesthetic induction with fentanyl. AB - The efficacy of fentanyl, 30 micrograms/kg, was evaluated as an anesthetic induction agent in 72 ASA I-III patients scheduled for 2-4-hr operations. The effect of preinduction pretreatment with pancuronium and/or diazepam and the incidence of loss of consciousness (anesthesia), recall, rigidity, abnormal muscle movements, and hemodynamic changes were documented. Seventy-four percent of all patients became anesthetized. Diazepam pretreatment enhanced but did not ensure success of anesthetic induction. There was a significant correlation between age and the incidence of unconsciousness (P = 0.0287) and all patients over 60 yr old were anesthetized with 30 micrograms/kg of fentanyl. The incidence and severity of rigidity was reduced by pancuronium (P = 0.0002) but not by diazepam pretreatment. However, pancuronium plus diazepam produced a significant reduction in the incidence of rigidity when compared to pancuronium alone (P = 0.031). A significant positive correlation between age and the incidence of rigidity (P = 0.003) was found. Six patients had focal and one patient global tonic-clonic abnormal muscle movements. Diazepam but not pancuronium significantly decreased both heart rate (P = 0.05) and blood pressure (P = 0.04). Seventeen patients required reversal of narcotic effect to restore adequate spontaneous respiration after surgery. No patient required postoperative mechanical ventilatory assistance. The results of this study demonstrate that 30 micrograms/kg of fentanyl is not a reliable anesthetic induction dose in patients less than 60 yr old. Both age and premedication enhance the anesthetic capabilities of induction with fentanyl.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981490 TI - Halothane-induced lipid peroxidation and glucose-6-phosphatase inactivation in microsomes under hypoxic conditions. AB - Halothane-induced lipid peroxidation was studied in microsomes from phenobarbital pretreated male rats at defined steady state oxygen partial pressures (PO2). At PO2 less than 10 mmHg on addition of halothane to NADPH-reduced microsomes, significant increases in malondialdehyde (MDA) formation, oxygen uptake, and conjugated dienes were measured. At the maximum, near a PO2 of 1 mmHg, halothane induced the formation of about 0.75 nmol MDA X mg microsomal protein-1 X min-1; it also stimulated microsomal oxygen uptake twofold to threefold, and caused an almost threefold increase in conjugated diene absorption. Moreover, at this PO2 microsomal glucose-6-phosphatase lost about 70% of its activity. At PO2 greater than 10 mmHg, no significant effects of halothane on MDA formation, oxygen uptake, conjugated diene absorption, and glucose-6-phosphatase activity were observed; likewise under anaerobic conditions there was only a slight increase in conjugated dienes. The findings demonstrate that halothane induces microsomal lipid peroxidation at low PO2 and in the presence of particular cytochrome P-450 isoenzymes, and that the halothane-induced lipid peroxidation leads to severe microsomal lesions, as indicated by the loss of glucose-6-phosphatase activity. PMID- 2981491 TI - Airway and cardiovascular responses to inhaled prostacyclin in normal and asthmatic subjects. AB - Prostacyclin (PGI2) is one of several prostanoids released after antigen challenge of human lung fragments. To define its activity on human airways, we studied the effect of inhaled PGI2 in 10 normal and 8 asthmatic subjects. In random order, PGI2, 1 mg/ml, its hydrolysis product, 6-oxo-PGF1 alpha, 1 mg/ml, and glycine vehicle were given on separate occasions by nebulizer. Measurements of specific airway conductance (SGaw), blood pressure (BP), heart rate (HR), plasma 6-oxo-PGF1 alpha, and cyclic AMP levels were made at frequent intervals for as long as 45 min after nebulization. Prostacyclin and 6-oxo-PGF1 alpha caused cough and retrosternal discomfort. None of the drugs had any significant effect on SGaw in either the normal or asthmatic subjects, though 2 asthmatics showed consistent bronchodilatation with prostacyclin. Prostacyclin caused a marked fall in diastolic blood pressure (mean 20 +/- 3 mmHg) and increase in heart rate (29 +/- 3 beats X min-1) with a small late fall in systolic blood pressure (8 +/- 2 mmHg). This was associated with a 12- to 15-fold increase in plasma 6-oxo-PGF1 alpha levels maximal at 1 min, and in normal subjects only, a later twofold increase in plasma levels of cyclic AMP maximal at 5 min. Thus, inhaled PGI2 at concentrations that had pronounced cardiovascular activity produced no consistent effect on airway caliber in normal or asthmatic subjects. PMID- 2981492 TI - Characteristics of pertechnetate movement across the canine tracheal epithelium. AB - Several 99mTechnetium-labeled markers have been used to study the barrier properties of the pulmonary epithelium; among these compounds is sodium pertechnetate (TcO4-). As TcO4- has pseudohalide properties, and as the characteristics of the movement of this ion across the airway epithelium are unknown, we examined its bidirectional permeability coefficients, and compared them with those of radiolabeled Cl- and mannitol in canine tracheal epithelium in vitro. We found that this ion behaves similarly to Cl- in its translocation across the airway epithelium. PMID- 2981493 TI - Neurologic dysfunction in the idiopathic hypereosinophilic syndrome. AB - Three specific patterns of neurologic deficit were seen in a group of 52 patients with the idiopathic hypereosinophilic syndrome. Central nervous system dysfunction was seen in 7 patients (15%), 4 of whom had a distinctive encephalopathy characterized by behavioral disturbances and upper motor neuron signs. Peripheral neuropathy was found in 27 patients (52%), with a sensory polyneuropathy the commonest form (in 23 patients), although mononeuritis multiplex and radiculopathy were also seen. Central nervous system abnormalities from embolic disorders were seen in 6 patients (12%). Thus, a total of 65% of these patients had some neurologic dysfunction. Although the spectrum of neurologic disease is broad and includes very different manifestations, we conclude that distinct patterns of neurologic involvement are characteristic of this syndrome. PMID- 2981494 TI - Chronicity of Epstein-Barr virus infection. PMID- 2981495 TI - Nitrofurantoin and peripheral neuropathy. PMID- 2981496 TI - Molecular epidemiology of cytomegalovirus infections associated with bone marrow transplantation. AB - Restriction endonuclease analysis of purified viral DNA was used to study the molecular epidemiologic characteristics of cytomegalovirus infection in 18 patients having bone marrow transplantation. Four patients who had had asymptomatic excretion of cytomegalovirus in urine before transplantation subsequently developed a cytomegalovirus infection after transplantation (pneumonia in 2 patients, fever and viremia in 1 patient, and asymptomatic viruria in 1 patient). In each patient, the infection that developed after transplantation was caused by a cytomegalovirus strain genetically identical to the isolate detected in urine before transplantation. Cytomegalovirus isolates from different sites (buffy coat, lung, and urine) of the same patient were also identical, but cytomegalovirus isolates from different patients were never identical. Our results suggest that some cytomegalovirus infections after bone marrow transplantation may be caused by strains present before transplantation. The great structural and genetic variability of cytomegalovirus isolates must be considered in the development of effective diagnostic and immunoprophylactic measures for infection after marrow transplantation. PMID- 2981498 TI - Treatment of Kaposi's sarcoma and thrombocytopenia with vincristine in patients with the acquired immunodeficiency syndrome. AB - Kaposi's sarcoma occurs in about one third of patients with the acquired immunodeficiency syndrome. Although in some patients the tumor is principally a cosmetic problem, other patients have progressive disease with significant morbidity. Twenty-three patients with Kaposi's sarcoma related to the acquired immunodeficiency syndrome were treated with vincristine. Three patients had a coexisting immune thrombocytopenia. Of the 18 patients evaluable for response, 11 had a partial response and 7 had a minor response. The median duration of partial response was 4 + months. All 3 thrombocytopenic patients developed a significant increase in platelet count, which in 2 was sustained with continued treatment for 6 and 9 months, respectively. We conclude that vincristine has antitumor activity in the epidemic form of Kaposi's sarcoma and that it is also effective in the treatment of associated immune thrombocytopenia. PMID- 2981497 TI - Apparent transmission of human T-cell leukemia virus type III to a heterosexual woman with the acquired immunodeficiency syndrome. AB - A 24-year-old woman developed the acquired immunodeficiency syndrome with lymphadenopathy, oral candidiasis, and Kaposi's sarcoma. Her only known risk factor for the syndrome was sexual contact with an asymptomatic Haitian man. The woman had serologic evidence for infection with human T-cell lymphotropic virus type III, and this virus was recovered from the saliva of her sexual partner. Epidemiologic and virologic studies of the cases of such patients provide further evidence of a primary pathogenetic role for this retrovirus in the acquired immunodeficiency syndrome. PMID- 2981499 TI - Pancreatic imaging. AB - Methods now available for evaluating the pancreas include computed tomography, ultrasound, angiography, endoscopic retrograde cholangiopancreatography, and percutaneous fine-needle aspiration. Computed tomography is best for initial evaluation of the patient with suspected pancreatic disorders as it is technically adequate and provides detailed anatomic information. Ultrasound also provides detailed information and survey capabilities but examinations are often technically unsatisfactory. Angiography, endoscopic retrograde cholangiopancreatography, and fine-needle aspiration provide important diagnostic information but are more invasive and require considerable operator skill. These techniques are secondary tests not indicated for the initial evaluation of every patient. No single test will always provide all necessary diagnostic information, and the appropriate utilization of each method requires an understanding of its strengths and weaknesses. PMID- 2981500 TI - Parvovirus infection and the acquired immunodeficiency syndrome. PMID- 2981501 TI - Rectal epithelium in the acquired immunodeficiency syndrome. PMID- 2981502 TI - Influence of ammonium ions on hepatic de novo pyrimidine biosynthesis. AB - Carbamyl phosphate (CP) is synthesized in the liver by two separate enzymes, CPS I and CPS II. CPS I, an intramitochondrial enzyme involved in ureogenesis, has a relative activity of 500- to 1000-fold greater than CPS II, a cytoplasmic enzyme which initiates the sequence of reactions for pyrimidine biosynthesis. The contributions of NH4Cl (substrate for CPS I) ang glutamine (substrate for CPS II) as precursors for pyrimidine biosynthesis in isolated hepatocytes were compared by measuring their effect on uracil nucleotide pool size, the incorporation of NaH14CO3 into these pools, and the accumulation of orotic acid. Physiological concentrations of NH4Cl caused a marked stimulation of incorporation of radioactivity into uracil nucleotides (6-fold increase at 0.5 mM NH4Cl), and radioactive orotate appeared in both the cells and the medium. In contrast, glutamine (at concentrations up to 10 mM) had no effect on the incorporation of radioactivity into uracil nucleotides, and no orotic acid was detected. Uracil nucleotide pools were expanded up to 50% by low levels of NH4Cl, but there was no expansion of this pool in the presence of added glutamine. NH4Cl-driven pyrimidine de novo biosynthesis was insensitive to feedback inhibition by an expanded uracil nucleotide pool, to galactosamine treatment, and to acivicin treatment, indicating that NH+4 stimulated pyrimidine biosynthesis as a result of CP synthesis by mitochondrial CPS I. The consequence of intramitochondrially produced CP being available for pyrimidine biosynthesis is that the controlling step of this pathway (CPS II) is bypassed. The appearance of orotic acid following NH4Cl stimulation indicated that the rate-controlling step of hepatic de novo pyrimidine synthesis under these conditions was orotate phosphoribosyl transferase. These data indicate that, at physiological concentrations of NH+4, the majority of uracil nucleotides synthesized in isolated rat hepatocytes was derived from intramitochondrially generated CP. The effect of NH4Cl on the output of uridine by the isolated perfused rat liver was examined. In the presence of a single addition of 20 mM NH4Cl, the excretion of uridine was increased from 100 200 to 375 nmol h-1 g-1 liver and orotic acid was released into the circulating perfusate reaching a maximum of 2 microM (in 220 ml of perfusate) after 2 h. With 40 mM NH4Cl, uridine export was increased to 450 nmol h-1 g-1 and a maximum of 5 microM orotic acid was released into the perfusate after 2 h.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981503 TI - Effect of deoxyribopolymers and ribopolymers on the sensitivity of the cyclic-AMP receptor protein of Escherichia coli to proteolytic attack. AB - The cAMP receptor protein (CRP) is an allosteric protein in which binding of cAMP effects a conformational change with a consequent increased affinity for DNA. Unliganded CRP is relatively resistant to attack by a variety of proteases (trypsin, subtilisin, Staphylococcus aureus V8 protease, clostripain, chymotrypsin) which cleave cAMP-CRP, producing N-terminal cores which have lost DNA binding activity. Binding of double-stranded deoxyribopolynucleotides and calf thymus DNA by cAMP-CRP confers protection against attack by trypsin, subtilisin, S. aureus V8 protease, and clostripain. Such cAMP-CRP-DNA complexes remain sensitive to attack by chymotrypsin. Of the single-stranded deoxy- and ribopolynucleotides tested, only r(I)n and r(A)n gave significant protection against attack by these proteases (with the exception of chymotrypsin). Since the cutting sites for trypsin (Lys 130) and subtilisin (Leu 116) are not part of the C-terminal DNA binding domain, it would appear that binding of DNA may confer conformational changes on other regions of cAMP-CRP. PMID- 2981505 TI - The substrate specificity, kinetics, and mechanism of glycerate-3-kinase from spinach leaves. AB - Glycerate-3-kinase (EC 2.7.1.31) from spinach leaves shows absolute specificity for D-glycerate as phosphate acceptor, yielding 3-phosphoglycerate as a product. ATP complexed with either Mg2+ or Mn2+ is the preferred phosphate donor. The enzyme has Km (D-glycerate) = 0.25 mM, Km (Mg-ATP) = 0.21 mM, Vmax = 300 mumol min-1 mg protein-1, and a turnover number = 12,000 X min-1. The equilibrium constant for the reaction is approximately 300 at pH 7.8. Pyrophosphate, 3 phosphoglycerate and ribulose 1,5-bisphosphate are the strongest inhibitors among the phosphorylated and nonphosphorylated metabolites tested; however, their regulatory role in vivo is questioned. Substrate kinetics, as well as product and analog inhibition data, are consistent with a sequential random mechanism. The distinct characteristic of the glycerate kinase-catalyzed reaction is the formation of a dead-end complex between the enzyme, D-glycerate, and 3 phosphoglycerate. PMID- 2981504 TI - Ca2+-stimulated phospholipid phosphoesterase activities in rabbit erythrocyte membranes. AB - The properties of the enzymes involved in Ca2+-stimulated breakdown of phosphatidylinositol 4'-phosphate (PIP), phosphatidylinositol 4',5'-bisphosphate (PIP2), and phosphatidic acid (PA) in rabbit erythrocyte ghosts were studied. At 25 degrees C, 1 to 180 microM Ca2+ rapidly stimulated the breakdown of PIP and PIP2, and maximal breakdown occurred within 10 minutes at all Ca2+ concentrations. The rate and the total amount of breakdown of PA, PIP, and PIP2 increased with Ca2+ concentration. MgCl2 inhibited the rate of Ca2+-stimulated breakdown of PIP and PIP2 at Ca2+ concentrations less than 10 microM, but did not have any appreciable effects at higher Ca2+ concentrations. MgCl2 also protected against Ca2+-stimulated breakdown of PA. In the presence and absence of 5 mM MgCl2, Ca2+ stimulated half-maximal breakdown of PIP and PIP2 at 2-3 microM under hypotonic and isotonic conditions. In the presence of 5 mM MgCl2, Ca2+-stimulated breakdown of PIP and PIP2 was associated with the release of Pi and inositol bisphosphate. In the absence of MgCl2, Ca2+ stimulated the release of 32P-labeled Pi, inositol bisphosphate, and inositol trisphosphate from labeled PIP, PIP2, and PA. Ca2+ increased phosphatidylinositol content and decreased PIP and PIP2 content in these membranes. The results of this investigation suggest that Ca2+ stimulates the breakdown of polyphosphoinositides by stimulating polyphosphoinositide phosphomonoesterase and phosphodiesterase activities in rabbit erythrocyte ghosts. These activities were activated by less than 3 microM Ca2+ in the presence of MgCl2 under hypotonic or isotonic conditions. These Ca2+ stimulated polyphosphoinositide phosphoesterase activities could therefore be active under physiological conditions in normal rabbit erythrocytes. PMID- 2981506 TI - Lipid photooxidation in erythrocyte ghosts: sensitization of the membranes toward ascorbate- and superoxide-induced peroxidation and lysis. AB - The damaging effects of ascorbate (AH-) and superoxide (O-2) on resealed erythrocyte ghosts containing predetermined levels of lipid hydroperoxides (LOOHs) have been studied. Continuous blue light irradiation of membranes in the presence of protoporphyrin resulted in steadily increasing LOOH levels and enhanced release of a trapped marker, glucose 6-phosphate (G6P), after a 3- to 4 h lag. Neither superoxide dismutase (SOD) nor catalase inhibited these effects, ruling out O-2 and H2O2 as reactive intermediates. A 1-h light dose produced partially photoperoxidized ghosts, which, in the dark at 37 degrees C, released G6P no faster than unirradiated controls (approximately 7%/h). When xanthine oxidase plus xanthine (XO/X) was introduced as a source of O-2 and H2O2, the irradiated membranes lysed rapidly (t1/2 approximately 2 h). EDTA or SOD inhibited the reaction, whereas catalase had little or no effect. Unirradiated ghosts were not lysed by XO/X unless the system was supplemented with Fe(III), in which case total protection was afforded by SOD or catalase. In all experiments there was an excellent correlation between postirradiation lipid peroxidation (thiobarbituric acid reactivity) and G6P release. Similar observations were made with AH-. For example, dark incubation of photooxidized ghosts in the presence of 0.5 mM AH- resulted in rapid lysis (t1/2 approximately 1 h), which was stimulated approximately twofold by 50 microM Fe(III) and was inhibited by EDTA. By comparison, unirradiated ghosts showed no net peroxidation or lysis after 3 h exposure to Fe(III)/AH-. Neither SOD nor catalase protected against AH- stimulated damage. AH--promoted lipid peroxidation was inhibited by butylated hydroxytoluene, a lipophilic antioxidant, but was unaffected by 2,5-dimethylfuran or ethanol, singlet oxygen, and hydroxyl radical traps, respectively. These results suggest that a mechanism exists by which photogenerated LOOHs undergo redox metal-mediated reduction to alkoxy radicals (LO.), which trigger a burst of membrane-disrupting lipid peroxidation. PMID- 2981507 TI - Dephosphorylation of L-pyruvate kinase during rat liver hepatocyte isolation. AB - In isolated rat liver cells, the inhibition of L-pyruvate kinase (L-PK) by a cyclic AMP-dependent phosphorylation mechanism is involved in the hormonal control of glycolysis and gluconeogenesis. The aim of this study was to ascertain whether or not the in vivo phosphorylation state of the enzyme was maintained during the liver perfusion used to prepare isolated liver cells. When the L-PK phosphorylation state was studied indirectly in liver extracts by kinetic measurement, it was found that, during the perfusion, the S0.5 of phosphoenol pyruvate (PEP) for L-PK was decreased in a time-dependent manner from 1 +/- 0.08 to 0.64 +/- 0.1 mM (P less than 0.01) and 0.58 +/- 0.06 mM in liver cells. This shift was prevented only by the addition of glucagon to the perfusion medium. The extent of phosphorylation of L-PK was also estimated by incubation of the liver extract with [gamma-32P]ATP, protein kinase, and cyclic AMP, and measurement of 32Pi incorporated in L-PK by specific immunoprecipitation. In liver extracts removed at the beginning of the perfusion, 0.4 mol Pi/mol L-PK was incorporated and there was no stimulation by cyclic AMP. In contrast, in the liver extracts removed after 30 min of perfusion, cyclic AMP stimulated 32P incorporation two to threefold, and 1.6 mol Pi/mol L-PK was incorporated. These data suggest that L-PK was activated by a dephosphorylation mechanism during rat liver perfusion. This phenomenon could be involved in the classical inactivation of gluconeogenesis observed in the perfused rat liver model. PMID- 2981508 TI - Purification of plasminogen activator from Rous sarcoma virus-infected chick embryo fibroblast culture medium. AB - A new procedure for the purification of plasminogen activator secreted by cultured Rous sarcoma virus-infected chick embryo fibroblasts was described. The enzyme was isolated from culture medium containing 0.75% calf serum depleted of plasminogen by lysine-agarose affinity column chromatography and of high molecular-weight protease inhibitors by ultracentrifugation. The culture conditions allowed convenient preparation of large amounts of culture fluid with relatively high concentrations of plasminogen activator. The purification of the enzyme was accomplished by affinity chromatography on fibrin-celite and p aminobenzamidine-agarose columns, and by gel-filtration chromatography in the presence of urea. The activity was recovered in greater than 90% yield, and the enzyme was essentially homogeneous when analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Yields from 500 ml culture fluid exceeded 500 micrograms. PMID- 2981509 TI - Kinetic aspects of the interaction of horse heart cytochrome c with sodium dodecyl sulfate. AB - The sodium dodecyl sulfate (SDS) concentration dependence of spectral changes in circular dichroism (CD) and in absorbance of cytochrome c were examined in the far-ultraviolet region, aromatic region, and the Soret band. The Soret peak obtained in 0.60 mM SDS was nine times greater than that of the native state. (The critical micelle concentration, CMC, of SDS was 2.2 mM in the phosphate buffer used.) The results indicated that the drastic change at the Soret band did not accompany the corresponding large-scale change in secondary structure of the protein. In the stopped-flow measurements, two and three processes were followed at 406 nm below and above the CMC, respectively. At 289 nm only one process was observed, and this corresponded to the second process at 406 nm. Therefore, the second process at 406 nm was considered to be a change in tertiary structure around the heme group. The first process and the third process seemed to reflect a change in the heme environment; the former appeared to be due to a solvent effect and the latter due to a binding effect of a large number of dodecyl sulfate ions. PMID- 2981510 TI - The nucleotide sequence of tyrosine tRNAQ* psi A from bovine liver. AB - The nucleotide sequence of tyrosine tRNAQ* psi A from bovine liver was determined to be pC-C-U-U-C-m2G-A-U-A-m2G-C-U-C-A-G-D-D-G-G-acp3U-A-G-A -G-C-m22G-m22G -A-G G-A-C-U-Q*-psi-A-m1G-A-psi m-C-C-U-U-A-G-m7G-D-m5C-G-C-U-G-G-T-psi-C-G-m1A -U-U-C C-G-G-C-U-C-G-A-A-G-G-A-C-C-AOH. This tyrosine tRNA is 76 nucleotides in length, and contains two hypermodified nucleosides--3 -3(3-amino-3-carboxylpropyl)uridine (acp3U) and beta-D-galactosylqueuosine (Q*). The molecule also has a pseudouridine in the middle position of the anticodon, and is the first tRNA sequenced which has an adjacent pair of N2,N2-dimethylguanosine (m22G) residues. PMID- 2981512 TI - Rat skeletal muscle glycogen synthase: phosphorylation of the purified enzyme by cAMP-dependent and -independent protein kinases. AB - Glycogen synthase was purified to near homogeneity from rat skeletal muscle, and was found to resemble the rabbit skeletal muscle enzyme in several respects. An apparent molecular weight (Mapp) of 86,000 was estimated from the electrophoretic mobility of the subunit on polyacrylamide gels in the presence of sodium dodecyl sulfate. Limited proteolysis of the rat synthase with trypsin resulted in the formation of species with MappS equal to 75,000, 69,000, and 67,000. The enzyme could be phosphorylated by cAMP-dependent protein kinase, phosphorylase kinase, and the cAMP-independent protein kinases, PC0.7 and FA/GSK-3. Essentially all of the phosphorylation observed occurred on serines located in two cyanogen bromide fragments, denoted CB-1 (Mapp = 13,000) and CB-2 (Mapp = 22,000). FA/GSK-3 and cAMP-dependent protein kinase phosphorylated sites in both fragments. Phosphate introduced by phosphorylase kinase was located exclusively in CB-1, and that incorporated with PC0.7 was found in CB-2. Phosphorylation by FA/GSK-3 reduced the electrophoretic mobility of the subunit, introduced heterogeneity into CB-2, and was synergistic with phosphorylation by PC0.7. To separate phosphorylation sites more completely, samples of glycogen synthase were subjected to extensive proteolysis using trypsin, followed by reverse-phase liquid chromatography. When phosphorylated by the same kinases, the pattern of fragments obtained with rat and rabbit skeletal muscle glycogen synthase were almost identical. The results presented provide strong evidence that the subunit of rat skeletal muscle glycogen synthase has at least five phosphorylation sites that are very similar, if not identical, to sites present on the rabbit muscle enzyme. PMID- 2981511 TI - Chromatium flavocytochrome c: kinetics of reduction of the heme subunit, and the flavocytochrome c-mitochondrial cytochrome c complex. AB - The kinetics of reduction of Chromatium vinosum flavocytochrome c heme subunit by exogenous flavin neutral semiquinones generated by laser flash photolysis have been investigated. Unlike the holoprotein, the isolated heme subunit was appreciably reactive with lumiflavin neutral semiquinone. The measured rate constant for the reaction (2.7 X 10(7) M-1 S-1) was comparable to those of c-type cytochromes having similar redox potentials. The ionic strength dependence of the reaction with FMN neutral radical indicated that the heme subunit had a small negative charge at the site of reduction. Taken together, these results suggest that the active site of the heme subunit is buried on complexation with the flavin subunit in the holoprotein. Horse cytochrome c formed a strong complex with Chromatium, but not Chlorobium, flavocytochrome c. Possible physiological electron acceptors such as HiPIP, cytochrome c', and cytochrome c-555 apparently did not bind to the flavocytochromes c. The rate constant for reduction by lumiflavin radical of horse cytochrome c complexed to flavocytochrome c was about twofold smaller than for reduction of horse cytochrome c alone. Flavocytochrome c was itself unreactive with exogenous flavin semiquinones. The ionic strength dependence of the reduction of the complex by FMN radical was also smaller than for horse cytochrome c in the absence of flavocytochrome c. Sulfite, which forms an adduct with the protein-bound FAD (FAD is bound in an 8-alpha-S-cysteinyl linkage), did not affect the reduction of horse cytochrome c in its complex with flavocytochrome c. We conclude that horse cytochrome c is reduced directly by exogenous flavins in its complex with flavocytochrome c, although the kinetics are slightly modified. These results are not unlike observations made with complexes of mitochondrial cytochrome c with cytochrome oxidase or cytochrome b5. PMID- 2981513 TI - [Significance of immunotherapy as an adjunct to surgical treatment in non-small cell carcinoma of the lung]. AB - Active and adoptive immunotherapy as an adjunct to surgical treatment in non small cell lung cancers was analysed, based on the results of our TF experiences and a literature review, and its significance was discussed. PMID- 2981514 TI - [Serum neuron-specific enolase as a marker of lung cancers]. AB - Neuron-specific enolase (NSE) in sera of lung cancer patients was studied in order to evaluate its clinical significance as a tumor marker. The subjects included 15 normal volunteers, 13 cases without malignant neoplasms or neuronal diseases and 42 lung cancer cases. NSE was quantified by a double antibody radioimmunoassay. As one of the sera from normal volunteers and control patients showed an NSE content 10 ng/ml or more, values of 10 ng/ml or over were considered to be positive. Seventeen of 42 sera from lung cancer patients showed positive NSE levels. Histological evaluation revealed that the degrees of NSE positiveness for small cell carcinoma, large cell carcinoma, squamous cell carcinoma and adenocarcinoma were 73%, 50%, 33%, and 21%, respectively, and that all the positive cases except for one were confined to disease stages III or IV. The level of NSE in patients with 10 ng/ml or more before surgery decreased to within normal limits 1-2 weeks after surgery Localization of NSE could be confirmed immunohistologically in small cell carcinoma cells. In conclusion, NSE was considered to be very useful as a tumor marker of the lung, especially in small cell carcinoma for diagnosis and determination of disease extent and response to therapy, and also in non-small cell carcinoma for the evaluation of treatment effectiveness. PMID- 2981515 TI - [Clinical value of postoperative chemotherapy for non-small cell lung cancer- with special reference to long-term combined chemotherapy combined with immunotherapy]. AB - The main reason for unfavourable surgical outcome of lung cancer is latent distant metastases over looked during surgery, which ultimately cause recurrence, or death of the patients even in cases undergoing curative surgery. This fact necessitates the indispensable use of systemic adjuvant therapy in patients under going surgery for lung cancer. There have been mary reports concerning the clinical efficacy of surgical adjuvant chemotherapy for non-small cell lung cancer using various kinds of drugs in various treatment modalities, but the results have been controversial. During the past eleven years, we have used postoperative chemotherapy in three ways over three different periods: in the earliest period, short-term combined chemotherapy (STCC) was used, in the middle period, intermittent long-term combined chemotherapy (ILTCC) was used in combination with immunotherapy for a randomized group, and in the latest period, when continuous long-term combined chemotherapy (CLTCC) with immunotherapy was employed. A comparison was then made between these three kinds of treatment groups. In Comparing of the results obtained for the earliest and middle periods, ILTCC showed a significantly improved beneficial effect over STCC in terms of further increased survival rate. Furthermore, by randomized study, it was clarified that the favourable effect of ILTCC was further improved by concomitant use of immunotherapy. CLTCC with immunotherapy carried out in the latest period seemed to be prevent early recurrences in patients with stage I or II who underwent curative surgery, even though a short-term observation period of for 20 months was employed. It is conceivable that the latest treatment modality used will exerted best the most favourable beneficial effect in comparison with the two early treatments. A review of the literature was presented along with a discussion of the clinical value of chemotherapy and immunochemotherapy as a surgical adjuvant. PMID- 2981516 TI - [Surgical therapeutic planning for non-small cell lung cancer]. AB - The survival rates of 380 resected cases of lung cancer in our hospital were analyzed according to curability and histological cell type. The overall 5-year survival rate for stage I a cases was 64.5%, that for stage I b 52.3%, and that for stage II 26.7%. However, there were distinct differences in survival rates between stages I a-II with mediastinal lymph node dissection and those without mediastinal lymph node dissection. Of these 380 tumors, many were advanced (for instance, stage III tumors comprised 180 cases). T3 tumors had better prognosis (40.7% showing 5-year survival) than N2 tumors (26.7% showing 5-year survival). Among stage III tumors, squamous cell carcinoma (T3: 41%, N2: 36.7% showing 5 year survival) had a better prognosis than adenocarcinoma (T3: 16.1%, N2: 21.4%). T3N2 tumors, however, had such a poor prognosis that the value of surgery in these cases seemed questionable. Adjuvant therapy should therefore be evaluated accurately in future to improve prognosis. It was stressed that a randomized controlled study would be needed to evaluate the effectiveness of adjuvant therapy. PMID- 2981517 TI - [Correlation between tumor histology and prognostic factors in lung cancer cases]. AB - The most important prognostic factor is TNM and related stage classification in lung cancer. As there are, nevertheless, many tumor cell types and growth patterns in lung cancer cases, more precise analysis of various morphologic features affecting prognosis are needed based on tumor histology. These include tumor location in squamous cell carcinoma, grades of scarring associated with a tumor, nuclear atypia of tumor cells, frequency of mitoses, immunologic responses of regional lymph nodes and degrees of infiltration of T-zone histiocytes in adenocarcinoma, and tumor cell subtypes in small cell and large cell carcinomas. Evaluating these prognostic factors according to histology at the time of surgery, it is easier to decide whether or not adjuvant therapy is necessary. PMID- 2981518 TI - Phenytoin modulates connective tissue metabolism and cell proliferation in human skin fibroblast cultures. AB - Phenytoin has been proposed for the treatment of certain dermatologic conditions involving connective tissue abnormalities. To understand the biochemical basis of connective tissue changes, we incubated human skin fibroblasts in culture with varying concentrations of phenytoin. The results indicated that fibroblast proliferation, detected by tritiated thymidine incorporation into cells, was slightly stimulated when short incubation periods and low concentrations of phenytoin were employed. However, with longer incubation times and higher phenytoin concentrations, a significant reduction in fibroblast proliferation was observed. Further studies demonstrated that incubation of cells with phenytoin did not affect the production of procollagen, measured as synthesis of radioactive hydroxyproline in the cultures. However, assay of prolyl hydroxylase, an enzyme participating in the post-translational synthesis of hydroxyproline during collagen biosynthesis, was significantly reduced in the fibroblast cultures. The activity of collagenase, an enzyme participating in degradation of collagen, was markedly decreased in cultures treated with phenytoin. Thus, phenytoin may modulate collagen metabolism primarily by affecting the degradation of collagen. The results support previous suggestions that phenytoin may be useful for treatment of patients with increased levels of collagenase, such as in recessive dystrophic epidermolysis bullosa. PMID- 2981519 TI - Increased natural killer cell activity in patients with epidermodysplasia verruciformis. AB - Six patients with epidermodysplasia verruciformis (EV) were studied for natural killer cell (NKC) cytotoxic response of their peripheral blood mononuclear cells against K-562 target cells in an 18-hour chromium 51-release assay. Four patients displayed higher cytotoxic responses than controls did, whereas two others did not differ from controls. Patients with EV who had increased NKC activity were found to be infected with potentially oncogenic human papillomaviruses (types 5, 8, 9, 14, 17, 19, 22, 24, and others), and they have had multiple skin carcinomas and/or Bowen's precancerous dermatosis. Two patients with EV who had normal cytotoxic response were free of skin malignancies. PMID- 2981521 TI - Distal sensory nerves of the lower extremity in peripheral neuropathy: comparison of medial dorsal cutaneous and sural nerve abnormalities. AB - This study investigates the sensitivity and usefulness of medial dorsal cutaneous nerve (MDCN) conduction studies in patients with peripheral neuropathy. Two hundred twenty-three patients with clinical signs and symptoms of peripheral neuropathy in their lower extremities were evaluated. Nerve conduction velocity studies of the MDCN were compared to those of the sural and peroneal motor nerves. Fifty-eight percent of the patients had no measurable sensory nerve action potential (SNAP) from either the MDCN or sural nerve or both, indicating moderate to severe neuropathy. Of the remaining patients with mild neuropathy, 12% had a normal sural nerve and an abnormal MDCN, whereas 7% had a normal MDCN and an abnormal sural nerve. Fifty-one percent of the mild cases had normal peroneal motor nerve studies. It is concluded that the MDCN is equal to the sural nerve as a sensitive indicator in all stages of peripheral neuropathy; in mild or early neuropathy approximately 12% of patients will have normal sural nerve studies; and it is important to examine both distal sensory nerves of the leg. Addition of the MDCN study to the standard sural study increases diagnostic accuracy in mild peripheral neuropathy from 88% to nearly 100%. PMID- 2981520 TI - Reoperation for persistent and recurrent hyperparathyroidism. AB - As a consequence of an experience with 175 patients undergoing reoperation for persistent or recurrent hyperparathyroidism, the authors have evolved an approach to management. The emphasis is on the confirmation of the diagnosis; the critical evaluation of invasive and noninvasive localization tests, techniques of intraoperative assessment as to the adequacy of the reexploration; the avoidance of postoperative complications; the treatment with cryopreserved autografts of persistent hypocalcemia. With this approach, the authors have had a success rate of greater than 90% for reexploration in patients with persistent and recurrent hyperparathyroidism. PMID- 2981522 TI - Myocardial dysfunction in septic shock. AB - Myocardial depression is a major but poorly understood component of septic shock. This study investigates the morphologic and biochemical abnormalities associated with septic shock. Myocardial cells are incubated in normal and septic plasma in a nutrient-, oxygen-, pH-, electrolyte-, and temperature-controlled environment. Cells and media are tested for basal- and epinephrine-stimulated cyclic adenosine monophosphate (cAMP), lactic dehydrogenase (LDH), and creatine kinase. Electron microscopic studies are done at the end of incubation. Septic LDH and creatine kinase levels in the media are increased substantially, and septic cAMP levels are reduced significantly. Septic cells beat irregularly and arrest along with exhibiting abnormal electron microscopic structure. Septic myocardial dysfunction occurs independently of previously postulated causes that are controlled for in this experiment and therefore may be due to endogenously produced or accumulated toxic factor(s). PMID- 2981523 TI - Inhibition of beta-lactamase-induced resistance in soft-tissue infections. AB - Sulbactam ([CP45,899] penicillanic acid sulfone) inhibits many of the beta lactamases commonly found to be the cause of penicillin resistance. This agent was combined with either penicillin G potassium or ampicillin sodium in the treatment of 97 patients admitted with serious soft-tissue infections. Fifty-one of the infections were caused by at least one bacteria resistant to the antibiotic alone. Staphylococcus aureus was the most common pathogen (48 isolations) followed by the coliforms (30 isolations). Ninety percent of the isolates that were tested produced beta-lactamase. Susceptibility studies showed a high degree of resistance to the penicillin alone that was significantly lowered by the addition of sulbactam. The overall clinical results showed 81% of the infections to be either well controlled or cured. Three patients failed to show improvement. Thirteen patients showed transitory increases relatively safe and efficacious in the treatment of soft-tissue infection caused by penicillin resistant and penicillin-susceptible organisms. PMID- 2981524 TI - Regulation of neutrophil superoxide production in sepsis. AB - Neutrophil superoxide production has been recognized as an important pathway for microbicidal activity and regulation of the local inflammatory environment. To investigate neutrophil superoxide production in sepsis, we studied 22 patients with intra-abdominal infections, and correlated superoxide production with chemotactic response and granular enzyme content. Our results showed that neutrophils from infected patients had specific loss of chemotactic response to C5a, and were deficient in the granular enzymes, lysozyme, and beta glucuronidase. Superoxide production in response to opsonized zymosan was intact, but response to the chemoattractant N-formyl-methionyl-leucyl-phenylalanine was markedly depressed. This could be reversed in vitro by the addition of cytochalasin B. These results suggest that down regulation of exocytosis of superoxide to nonphagocytic stimuli occurs during sepsis, possibly protecting the host from tissue injury due to oxide radical release. Superoxide response to phagocytic stimulation was intact. PMID- 2981525 TI - Use of a malar bone graft to augment skull-base access. AB - An anterolateral transfacial approach for tumors of the retromaxillary space, the nasopharynx, and the middle cranial fossa features a temporary removal of the malar complex and resection of the coronoid process and the pterygoid plates. Two cases demonstrate use of this approach. Results and complications are observed in six patients. PMID- 2981526 TI - Dynamic computed tomography. Its use in the assessment of vascular malformations and angiofibroma. AB - Reliable diagnostic methods for head and neck vascular lesions are needed. The technique of dynamic computed tomography (CT) is used to delineate two aberrant subclavian arteries, one carotid artery aneurysm, and one angiofibroma. Five or six 4.8-s CT scans were obtained sequentially, separated by a 1.2-s interscan interval. Computer-generated iodinated contrast concentration (CT number) v time curves showed rapid influx of contrast, high peak density, and rapid uninterrupted contrast runoff. Simultaneous bone and soft-tissue visualization at the time of peak contrast enhancement provides excellent delineation of the malformations and angiofibroma. The technique is noninvasive, and it eliminates the potential complications of arteriography and biopsy. We conclude that dynamic CT is an excellent method for diagnosis of head and neck vascular lesions. PMID- 2981528 TI - Receptor binding and biological activity of bivalent enkephalins. AB - Two series of dimeric enkephalin analogues were assayed for opioid activity in two isolated smooth muscle preparations: the guinea pig ileum (GPI) and the mouse vas deferens (MVD). Dimers have the general structure: X-(CH2)n-X, where X is H Tyr-D-Ala-Gly-Phe-Leu-NH-(n = 0, 2, 4, 6, 8, 10, 12), for the first series of dimeric pentapeptide enkephalins (DPEn), and H-Tyr-D-Ala-Gly-Phe-NH-(n = 2, 4, 6, 8, 12), for the series of dimeric tetrapeptide enkephalins (DTEn). Comparison of biological activities with binding affinities revealed that: (1) the DPE series with n = 2-8 showed increased potency in the MVD assay relative to monomeric [D Ala2, Leu5]enkephalinamide (DALEA); (2) there was an associated increase affinity for the delta receptor of rat brain or neuroblastoma-glioma hybrid cells. (however, the relative potencies were higher in the MVD assay then predicted on the basis of binding affinities); (3) the DTE series also showed an increase in delta receptor affinities and MVD potencies relative to DALEA, for n = 2-12; (4) for the DTE series, the increase in MVD activities was less than that expected on the basis of delta binding affinity; (5) for both the DPE and DTE series, activities in the GPI assay and mu-receptor affinities were highly correlated: as the length of the methylene bridge increased from 2 to 12, there was a progressive loss of activity in both assays, with a similar pattern for DPE and DTE. Two selected dimers and their corresponding monomers were also assayed for antinociceptive activity in vivo: results were consistent with GPI and mu-binding but not with MVD and delta-binding. Two alkylamide analogs of penta- and tetrapeptide monomers, representing the monomer with the attached spacer of the most active dimers, were also assayed in biological and binding assays. Comparison of these compounds with the corresponding dimers suggest that the changes in activities and selectivities induced by dimerization are not a spurious effect of the presence of an akylamide derivative of the carboxy terminal of enkephalin but rather may represent a specific effect due to the bivalent nature of the ligands. PMID- 2981527 TI - Platelet activation. AB - Platelets are discoidal cytoplasmic particles that respond to a variety of stimuli by developing filopodia and rounding up (shape change), developing the ability to bind fibrinogen from the medium, and, with strong stimuli such as thrombin and PAF-acether, secreting the contents of several types of granules. Arachidonic acid is cleaved from phospholipids by phospholipase A2 and converted by the platelets to endoperoxides, and then to thromboxane A2. The bound dimeric fibrinogen molecules probably cause aggregation by forming bridges between platelets. Aggregation is reinforced by secreted fibrinogen and thrombospondin, which binds the platelets, and by thromboxane A2 and endoperoxides, as well as secreted ADP, which cause additional receptor-mediated activation. The responses to these stimuli are initiated when the agonists bind to specific receptors on the plasma membrane. Subsequent steps resemble those in other types of responsive cells: breakdown of phosphatidylinositol bisphosphate into diacylglycerol, a stimulator of protein kinase C, and inositol-1,4,5-trisphosphate, recently shown to be a potent calcium ionophore. The response of shape change results from increased cytoplasmic Ca2+ which permits phosphorylation of one of the light chains of myosin by a calcium-calmodulin-dependent kinase, with resulting enhanced actin-myosin interaction. Secretion is associated with phosphorylation of a 40,000 to 47,000 dalton protein by the diacylglycerol-activated protein kinase C. These recent findings have increased our understanding of the mechanisms of platelet activation, but much remains to be learned. How do agonist receptor complexes influence PIP2 breakdown? Is this indeed the first step in activation? What mediates adhesion of platelets to the injured blood vessel wall? Does transduction of this stimulus occur by the same mechanism as transduction of commonly used soluble stimuli? What is the role of the phosphorylated 40-47 K protein in secretion? What change in GP IIb-IIIa promotes their ability to bind fibrinogen? What is the role of calcium-activated protease? Of the phosphorylation of actin-binding protein? Progress is being made rapidly, and these questions may be answered within a few years. PMID- 2981529 TI - Entrapping of the spin label tempocholine into human erythrocytes by resealing after hyposmolar stress. Comparison with haemolysis. The effects of some membrane active substances. AB - Human erythrocytes were subjected to a sudden hyposmolar stress by suspension in solutions of varying salt concentrations in the presence of the spin label tempocholine. The enlarged pores in the erythrocyte membranes produced by the influx of water, followed by stretching, allowed the passage of the spin label, so that a certain amount of tempocholine was entrapped when the erythrocytes spontaneously resealed with closing of the pores. The excess of spin label in the external solution was then reduced to a diamagnetic species by the addition of ascorbic acid. The positively charged tempocholine and the ascorbic acid did not penetrate properly resealed erythrocytes, so that the electron spin resonance (ESR) signal from the entrapped spin label constituted a measure of the effective resealing of the pores and rifts in the membrane subsequent to hyposmolar stress. Some drug substances were found to influence the entrapping curves obtained when the amount of entrapped spin label was plotted against the osmolarity. Chlorpromazine, trifluoperazine, nicardipine, amperozide and haloperidol gave rise to a dose-dependent decrease of the entrapping of tempocholine, especially at low osmolarities. The exclusion of Ca2+ and Mg2+ ions from the solutions increased the action of chlorpromazine. The protective action against haemolysis brought about by a number of membrane-active substances at low concentrations [2] had its counterpart in the entrapping curve observed with chlorpromazine at 0.1 mM. It is suggested that the substances in this series exerted their action on the resealing process by interaction with the calmodulin system. PMID- 2981530 TI - Structural changes of rat liver microsomal membranes induced by the oral administration of carbon tetrachloride. 31P-NMR and spin-label studies. AB - The acute effects of carbon tetrachloride (CCl4) on the membrane structure of rat liver microsomes were studied using 31P-NMR and spin-labeling techniques. 31P-NMR spectra of rat liver microsomes were not changed appreciably after the oral administration of CCl4, indicating that the surface structures of microsomal membranes probably are not influenced by the oral administration of CCl4. Four different spin-labeled stearic acids, 5-(N-oxyl-4',4'-dimethyloxazolidine) stearic acid (5SLS), its methyl ester (5SLSM), 12-(N-oxyl-4',4' dimethyloxazolidine)-stearic acid (12SLS) and its methyl ester (12SLSM), were used for the estimation of membrane fluidity. The apparent rotational correlation time of 12SLS decreased from 4.0 nsec to 3.0 nsec after the oral administration of CCl4, while the order parameter of 5SLS did not change. The results suggest that CCl4 or its metabolites increase the membrane fluidity of liver microsomes primarily at hydrophobic regions rather than at the surface layer. The ESR spectrum of 5SLSM in microsomal membranes comprised two different signals; one was an anisotropic signal and the other was a rather isotropic one. The ratio of the anisotropic signal to the isotropic one decreased markedly after the oral administration of CCl4 and depended on the dose of CCl4. The suitability of this ESR technique with 5SLSM for the estimation of membrane damage is discussed. PMID- 2981531 TI - Effects of cycloheximide and tunicamycin on opiate receptor activities in neuroblastoma X glioma NG108-15 hybrid cells. AB - The molecular mechanism of opiate receptor down-regulation and desensitization was investigated by studying the effects of cycloheximide and tunicamycin on opiate receptor activities in neuroblastoma X glioma NG108-15 hybrid cells. Cycloheximide inhibited [35S]methionine and [3H]-glucosamine incorporation by hybrid cells, while tunicamycin inhibited [3H]glucosamine incorporation only. Exposing hybrid cells to these two agents did not alter the viability of the cell. Treatment of NG108-15 cells with cycloheximide or tunicamycin produced a decrease in [3H]diprenorphine binding dependent on both time and concentrations of inhibitors, with no measurable modification in the ability of etorphine to regulate intracellular cyclic AMP production. Cycloheximide attenuated [3H] diprenorphine binding by decreasing both the number of sites, Bmax, and the affinity of the receptor, Kd. Tunicamycin treatment produced a decrease in Bmax with no apparent alteration in Kd values. Cycloheximide and tunicamycin did not potentiate the rate or magnitude of etorphine-induced down-regulation or desensitization of opiate receptor in NG108-15 cells. Furthermore, there was an apparent antagonism in cycloheximide action on receptor down-regulation. The reappearance of opiate binding sites after agonist removal was affected by these two inhibitors. Cycloheximide and tunicamycin eliminated the increase in [3H]diprenorphine binding in the chronic etorphine-treated cells after agonist removal. These two inhibitors did not alter the resensitization of hybrid cells to etorphine. Thus, the site of opiate agonist action to induce receptor down regulation and desensitization is not at the site of protein synthesis or protein glycosylation. These data substantiate previously reported observations that receptor down-regulation and receptor desensitization are two different cellular adaptation processes. PMID- 2981532 TI - Characterization of solubilized "peripheral type" benzodiazepine binding sites from rat adrenals by using [3H]PK 11195, an isoquinoline carboxamide derivative. AB - "Peripheral type" benzodiazepine binding sites have been solubilized with digitonin. Binding site density for the solubilized material is increased 1.7 times compared to membranes. A decrease in the affinity for [3H]-PK 11195 (a new ligand for the peripheral type benzodiazepine binding sites) was also observed. Pharmacological specificity of displacing agents was conserved during solubilization. The apparent molecular weight determined by gel filtration was 215,000 +/- 20,000. The high Bmax value of the solubilized preparation (greater than 50 pmole/mg protein) makes it advantageous as the starting point for a purification procedure. PMID- 2981533 TI - Effects of dihydropyridine calcium channel blocking drugs on rat brain muscarinic and alpha-adrenergic receptors. AB - The dihydropyridine (DHP) Ca2+ channel blocking drugs nicardipine, nitrendipine, nimodipine, felodipine, nifedipine and nisoldipine were examined for activity in inhibiting specific (-)-[3H] QNB and [3H]WB4101 binding to the muscarinic and alpha-adrenergic receptors, respectively, of rat brain. Muscarinic receptor binding was affected most by nicardipine, with felodipine having less activity; the other DHP drugs were essentially inactive at 3 X 10(-5) M. The (+) stereoisomer nicardipine (KI = 4.07 X 10(-7) M) was 27 times more potent than the (-)-isomer in inhibiting [3H]QNB binding, and this inhibition was found to be competitive. This inhibitory effect of nicardipine was not mediated via interaction with the high-affinity DHP binding site assumed to be associated with a Ca2+ channel. (+)-Nicardipine inhibited the binding of [3H]nitrendipine to this DHP binding site of brain, with a K1 of 9.01 X 10(-11) M, and was 10 times more potent than the (-)-isomer. Thus, the muscarinic receptor was 4200 times less sensitive to (+)-nicardipine than was this DHP binding site. Nicardipine was also the most potent DHP drug inhibiting [3H]WB4104 binding to the alpha-adrenergic receptor, although the other drugs were also somewhat active, in the rank order sequence listed above. This effect of nicardipine on the adrenergic receptor was also stereoselective, with (+)-nicardipine (KI = 3.46 X 10(-7) M) being about 3 times more potent than the (-)-isomer, in producing competitive inhibition of radioligand binding. These data suggest that the effects on brain receptors occur as a result of direct, stereospecific effects of DHP drugs on these receptors and are not due to Ca2+ channel blocking activity of these drugs. PMID- 2981534 TI - Relative inhibitory potency of five mineralocorticoid antagonists on aldosterone biosynthesis in vitro. AB - Spirolactones are mineralocorticoid antagonists which bind to aldosterone receptors in the distal nephron. During the last decade, several antimineralocorticoids, which are more potent than spironolactone in competing for mineralocorticoid receptors have been developed. In the present study, we have compared the direct activity of spironolactone and four related compounds: prorenone (SC 23133), SC 19886, SC 26304 and its carboxylic analog SC 27169, on aldosterone biosynthesis. Two of them (SC 26304 and its carboxylic analog SC 27169) had no effect on adrenal steroidogenesis, even at concentrations up to 10( 3)M. Spironolactone and prorenone (SC 23133) induced a marked but reversible inhibition of aldosterone biosynthesis. SC 19886 totally inhibited aldosterone production and the activity of this compound lasted for more than 7 hours. In addition, SC 19886 and prorenone (SC 23133) totally suppressed ACTH and angiotensin II-induced stimulation of aldosterone biosynthesis whereas SC 27169 was unable to block adrenal response to these corticotropic hormones. Our results suggest that compounds such as prorenone (SC 23133), SC 19886 and spironolactone, which are potent inhibitors of aldosterone biosynthesis could be more active in the treatment of primary aldosteronism than those antimineralocorticoids which are devoid of action on aldosterone biosynthesis. PMID- 2981535 TI - Enhanced oxygen consumption in adrenal medulla on stimulation with acetylcholine. AB - When perfused cortex-free ox adrenal medulla was stimulated to secrete catecholamine by infusion of 0.1 mM acetylcholine for 4 min, the oxygen consumption increased to a value which was 0.15 +/- 0.07 mumole O2/min/g wet weight (+/- S.D., N = 12) above the pre-stimulation value of 0.49 +/- 0.15 (P less than 0.001). 1.4 +/- 0.9 (+/- S.D., N = 12) mole of catecholamine was secreted per mole of enhanced O2 consumption in the 16 min following the start of the stimulation. The rate of ATP hydrolysis by the proton-translocating Mg-ATPase of the chromaffin granule may increase on fusing with the plasma membrane of the chromaffin cell during exocytosis. However, from the amount of catecholamine secreted, this was estimated to account for less than 17% of the oxygen consumption increase. The amount of catecholamine secreted by 4 min 0.1 mM acetylcholine stimulations correlated with the enhancement of oxygen consumption (r = 0.82, P less than 0.001) but, on stimulation with 60 microM veratridine for 4 min, O2 consumption enhancement was anomalously low. This dependence on mode of stimulation suggests that ATP consumption in exocytosis itself is an inadequate explanation. Ouabain-sensitive oxygen consumption rose from undetectable levels to 18 +/- 8% (+/- S.D., N = 4) of the basal respiration during prolonged 0.1 mM acetylcholine stimulation in the absence of Ca, indicating that Na,K-ATPase was not responsible for all of the oxygen consumption enhancement. PMID- 2981536 TI - Use of desferrioxamine as a 'probe' for iron-dependent formation of hydroxyl radicals. Evidence for a direct reaction between desferal and the superoxide radical. AB - The iron chelator desferal is a powerful inhibitor of lipid peroxidation and of hydroxyl radical formation dependent on the presence of iron salts. Desferal also reacts with superoxide radical with a second-order rate constant approximately equal to 3 X 10(2) M-1 s-1 at pH 10.2 and approximately 9 X 10(2) M-1 s-1 at physiological pH. It is concluded that this slow reaction of desferal with O2- is unlikely to influence the interpretation of experiments in which the chelator is used. The ability of desferal to react with hydroxyl radical (k2 approximately 10(10) M-1 s-1) is a far more likely source of error in the interpretation of results using this chelating agent. PMID- 2981537 TI - Chondrocytes that accumulate proteoglycans and inorganic pyrophosphate in the pathogenesis of chondrocalcinosis. PMID- 2981538 TI - "Concomitant immunity" in murine tumours of non-detectable immunogenicity. AB - Various immunization assays were used to demonstrate the lack of immunogenicity of three BALB/c tumours of spontaneous origin and of a fourth one resulting from foreign body tumorigenesis. All four tumours inhibited the growth of a second implant of the same tumour into the contralateral flank. In our tumour models "concomitant immunity" (1) was not mediated by macrophage or T-cell dependent immune reactions: both thymectomized BALB/c and nude mice (treated or untreated with silica) gave the same results as intact mice; (2) showed some degree of non specificity, inhibiting the growth of a different tumour in 3/4 cases; though, the existence of a specific component could not be discarded; (3) was proportional to the volume of the primary tumour at the time of the second challenge; (4) was dependent on actively growing primary tumour, not being obtained with progressively increasing daily inocula of irradiated tumour cells; (5) was detectable in an actively growing secondary tumour; recurrent growth after partial surgical excision was inhibited and (6) involved cytostasis of the secondary tumour: a syngeneic graft of the overlying skin led to tumour growth while histological studies revealed the presence of viable tumour cells. It is postulated that "concomitant immunity" or resistance can be generated without the active participation of the immune system and that tumour-related factors are, in certain cases, responsible for blocking the growth of secondary tumours. PMID- 2981539 TI - HNK-1+ cells in non-Hodgkin's lymphoma: lack of relation with transferrin receptor expression on malignant cells. AB - It has been proposed that Natural Killer (NK) cell activity is involved in host defence against neoplasia, and that NK cells react with or recognize the transferrin receptor (TrR) on target cells. HNK-1 expression has been related to NK cell function. Therefore, in 118 cases of non-Hodgkin's lymphoma (NHL) we studied the occurrence and distribution of HNK-1+ cells by immunohistochemistry, and simultaneously assessed the expression of TrR on malignant cells. In NHL of intermediate or high grade malignancy there was uniform expression of TrR on malignant cells. In low grade malignancy NHL, only lymphocytic and lymphoplasmacytoid lymphomas were TrR negative, except for faint staining of proliferation centres. In 23 cases of follicular lymphoma, 9 showed the absence of HNK-1+ cells in neoplastic follicles. In 16/23 cases HNK-1+ cells were present around follicles or in interfollicular areas: 8 of these cases revealed a higher density of HNK-1+ cells at this site than inside the follicles. In 22/26 cases with high grade malignancy NHL, HNK-1+ cells were absent or present in small density, which is different from the presence in higher density in low grade malignancy NHL. We conclude that (i) TrR expression on NHL cells is not obligatory related with histological class or malignancy grade of the tumour, and that (ii) HNK-1+ cells are not universally present in areas of malignant cells, in particular in follicular lymphoma and in NHL of high grade malignancy. PMID- 2981540 TI - Growth of human bronchial carcinomas in nude mice. AB - Two hundred and thirteen lung tumours of primary site and 42 metastases were heterotransplanted into nude mice with an overall success rate of 44%. There were differences in success between the histological types. Squamous cell and adenocarcinoma had the highest success rate (51% and 43%, respectively) whereas large cell and small cell carcinoma had a lower success rate (38% for both). The average volume doubling times in the first passage in nude mice ranged from 8.2 in large cell carcinomas to 18.9 days in adenocarcinomas. In subsequent passages an increase in growth rate was found, the overall average doubling time falling from 14.5 days in the first passage to 7.1 days in the second passage. In a study with 171 non-small cell lung carcinomas (NSCLC), the growth data in nude mice were correlated with the clinical data of the corresponding patients. A relationship between the growth parameters in nude mice and prognosis of patients could not be found. PMID- 2981541 TI - Multipotential behaviour of cloned rat mesothelioma cells with epithelial phenotype. AB - Reference cultures derived from a transplantable rat mesothelioma were obtained by cloning cells three times in soft agar. Each line, designated "CARM-Lines", was selected on the basis of their epithelial or fibroblastic phenotype, and their uniform morphology. Three epithelial lines were used for more detailed in vitro studies comparing morphological and biological criteria at early and late passages. All three lines exhibited both epithelial and fibroblastic elements after 10-14 passages in vitro, demonstrating that the dimorphic histology of these tumours could be derived from a single aberrant cell. Morphology and growth characteristics of these cells were density-dependent. Anchorage dependent and independent clonogenic assays did not correlate. Anchorage dependent colony formation was the only parameter which differed markedly from the original parent line in the assays described. In vivo evidence of chondrogenesis and attempted ossification support the concept of a multipotential cell contributing to the diverse primary tumour morphology by cellular modulation or differentiation. PMID- 2981542 TI - Percoll density gradient separation of cells from human malignant effusions. AB - A simple method is described for the separation of cells derived from effusions of patients with adenocarcinomas in discontinuous density gradients of Percoll. After separation, cells from different fractions were analyzed by morphologic, histochemical and immunologic criteria. Total cell recovery from 27 experiments was 67 +/- 4%. Macrophages (82%) were recovered in the intermediate density fraction (1.056-1.067 g ml-1) with a purity of 90%. Recovered lymphocytes (98%) were found in the high density fraction (1.067-1.077 g ml-1) with a purity of 92%. The majority of the lymphocytes recovered were T cells. Malignant adenocarcinoma cells (90%) were recovered in the lowest density fractions (up to 1.056 g ml-1) with a purity of 79%. Use of effective cell separation procedures should facilitate the analysis of the functional capacities of both normal and neoplastic cells derived from human malignant effusions. PMID- 2981543 TI - Isolation and characterization of soluble cytochromes, ferredoxins and other chromophoric proteins from the halophilic phototrophic bacterium Ectothiorhodospira halophila. AB - A cytochrome c-551 and a pair of 'high redox-potential' ferredoxins (iso-high potential iron-sulfur proteins) were found to be the major soluble electron transport proteins in Ectothiorhodospira halophila. Smaller amounts of 'bacterial' ferredoxin and cytochrome c' were also observed. With the exception of cytochrome c-551, these proteins are commonly encountered in the purple sulfur bacteria, family Chromatiaceae and less frequently in the purple bacteria, family Rhodospirillaceae. In addition to the cytochromes and ferredoxins, E. halophila synthesizes substantial amounts of a small yellow-colored protein, which has a chromophore spectrally similar to flavins having oxygen, nitrogen or sulfur substituents in place of the 8-methyl group such as roseoflavin and the methanogen cofactor F-420. A purple-colored protein was only partially purified, but it is spectrally similar to iron proteins having a tyrosine ligand, such as transferrin, catechuate dioxygenase, and especially the purple acid phosphatases. Neither the yellow protein nor the purple one has previously been observed in phototrophic bacteria, but may in some way be required for survival in extremely halophilic habitats. The only feature common to halophiles including E. halophila is the very acidic nature of their proteins. PMID- 2981544 TI - Chemical modification of the Na+/H+ exchanger of thymic lymphocytes. Inhibition by N-ethylmaleimide. AB - A Na+/H+ exchanger is involved in the regulation of cytoplasmic pH and cellular volume in a variety of cells. Little is known about the molecular nature of this exchanger. The purpose of this study was to survey a variety of group-specific covalent reagents as potential inhibitors of the exchanger. Na+/H+ countertransport activity was assayed as the amiloride-sensitive rate of Na+ induced alkalinization in acid-loaded lymphocytes, or as the rate of swelling in cells suspended in sodium propionate medium. Activity was not affected by proteinases or by carboxyl-group and amino-group specific reagents. A significant inhibition was produced by diethylpyrocarbonate, a histidine-specific reagent and by N-ethylmaleimide, a sulfhydryl group reagent. A similarly reactive but nonpermeating sulfhydryl agent, glutathione-maleimide, failed to inhibit Na+-H+ exchange. Moreover, the reaction with N-ethylmaleimide was sensitive to changes in the cytoplasmic pH. The data suggest that the chemically reactive groups of the Na+/H+ exchanger of lymphocytes have limited exposure to the extracellular medium but that an internally located sulfhydryl group is critical for the cation exchange activity. PMID- 2981545 TI - Multiequilibrium binding of a spin-labeled local anesthetic in phosphatidylcholine bilayers. AB - The equilibria among spin-labeled amine local anesthetic species in dioleoylphosphatidylcholine liposomes at an anesthetic: lipid mole ratio of 1:100 are investigated. Electron spin resonance (ESR) spectra demonstrate that anesthetic mobility within the bilayer is charge-dependent, with the uncharged species the more mobile. Partition coefficient measurements confirm ESR evidence that changes in anesthetic mobility represent anesthetic-phospholipid interaction and not changes in bilayer fluidity. Spin-exchange attenuation experiments show that anesthetics within the bilayer are accessible to the aqueous medium. Dependence of tertiary-amine anesthetic pK on dielectric constant has been used to estimate the interfacial pK. We propose a model of equilibria among species of the tertiary amine anesthetic in the aqueous medium and those intercalated in the bilayer, including a species electrostatically bound to the lipid phosphate. Using experimentally determined equilibrium constants, the model provides the binding constant between the electrostatically bound and unbound cationic anesthetics within the bilayer. The model stimulates the pH dependence of the mobile fraction of total anesthetic population determined by subtraction techniques on experimental ESR spectra. PMID- 2981546 TI - Spectroscopic characterization of vesicle formation on heated human erythrocytes and the influence of the antiviral agent amantadine. AB - EPR investigations on the vesiculation process of heated human erythrocytes were performed, using different fatty acid spin labels. Spectrin denaturation and vesiculation do not influence the fluidity of the lipid phase of the remaining membrane of human erythrocytes: Vesicles released differ in chemical composition as well as in the lipid fluidity of their membrane from the intact human erythrocyte membrane. A reduced cholesterol-to-phospholipid ratio and a depletion of spectrin was found. By changing the ionic concentration of the suspension medium an effect on membrane spectra and on vesicle release was established. The adamantane derivative amantadine causes fluidization of the human erythrocyte membrane and inhibits vesicle release. Based on these results, a model for the mechanism by which adamantane-like molecules could interact with membranes is proposed. PMID- 2981547 TI - Affinity targeting of Sendai virions to desialized human erythrocytes using hybrid antibody molecules. AB - F(ab') fragments obtained from anti-Sendai virus antibodies were chemically coupled to F(ab') fragments obtained from anti-human red blood cell antibodies (anti-hRBC-Ab). This led to the formation of hybrid antibody molecules (anti-SV anti-hRBC(F(ab')2) each of whose F(ab') fragment possessed different binding specificity. The anti-SV(F(ab'] part of the hybrid molecule interacted specifically with Sendai virus particles, while the anti-hRBC(F(ab'] part interacted with the surface of hRBC. These hybrid antibodies were able to mediate binding and fusion of SV to hRBC, from which the virus receptors were removed by treatment with neuraminidase (desialized hRBC). Neither anti-SV-anti-SV(F(ab')2) nor anti-hRBC-anti-hRBC(F(ab')2) possessed the same ability. Thus, it is shown that soluble, hybrid antibody molecules can effectively mediate functional binding of Sendai virus to virus-receptor-depleted cells. PMID- 2981549 TI - A fluorescence quenching technique for the measurement of paramagnetic ion concentrations at the membrane/water interface. Intrinsic and X537A-mediated cobalt fluxes across lipid bilayer membranes. AB - We have characterized the quenching of N-(7-nitrobenz-2-oxa-1,3-diazol-4 yl)phosphatidylethanolamine by Co2+ in egg phosphatidylcholine (PC) lipid bilayer vesicles. The quenching constant obtained is 59 M-1. We demonstrate one use of this fluorescence quenching technique by measuring intrinsic and X537A-mediated transmembrane Co2+ fluxes in large unilamellar PC vesicles. The intrinsic rate constant for Co2+ flux we measure is 3 X 10(-6) S-1. We confirm that the neutral Co approximately (X537A)2 complex is the main component of the X537A-mediated cobalt flux. Since this method measures the concentration of Co2+ at the site of the fluorophore, it is generally applicable to the measurement of paramagnetic ion concentrations in the region of the membrane/water interface. PMID- 2981548 TI - Na+-stimulated ATPase activities in basolateral plasma membranes from guinea-pig small intestinal epithelial cells. AB - Two ATPase activities, a Na+-ATPase and a (Na+ + K+)-ATPase, have been found associated with sheets of basolateral plasma membranes from guinea-pig small intestinal epithelial cells. The specific activity of the former is 10-15% of the latter. The two ATPase activities differ in their affinity for Na+, their optimal pH, their K+ requirement and particularly in their behaviour in the presence of some inhibitors and of Ca2+. Thus the Na+-ATPase is refractory to ouabain but it is strongly inhibited by ethacrynic acid and furosemide, whilst the (Na+ + K+) ATPase is totally suppressed by ouabain, partially by ethacrynic acid and refractory to furosemide. In addition, the Na+-ATPase is activated by micromolar concentrations of calcium and by resuspension of the membrane preparation at pH 7.8. The Na+-ATPase is only stimulated by sodium and to a lesser extent by lithium; however, this stimulation is independent of the anion accompanying Na+. The latter rules out the participation of an anionic ATPase. The relation between the characteristics of the sodium transport mechanism in basolateral membrane vesicles (Del Castillo, J.R. and Robinson, J.W.L. (1983) Experientia 39,631) and those of the two ATPase activities present in the same membranes, allow us to postulate the existence of two separate sodium pumps in this membranes. Each pump would derive the necessary energy for active ion transport from the hydrolysis of ATP, catalyzed by different ATPase systems. PMID- 2981550 TI - In vivo interaction between nitrogenase molybdenum-iron protein and membrane in Azotobacter vinelandii and Rhodospirillum rubrum. AB - Oriented whole cell multilayers of Azotobacter vinelandii and Rhodospirillum rubrum were analyzed by electron spin resonance (ESR) spectroscopy to detect possible structural associations between nitrogenase molybdenum-iron (MoFe) protein and cytoplasmic or intracytoplasmic membrane. Initially, protocols were designed to obtain strong molybdenum-iron protein ESR signals in whole cell samples of each organism. Then, two-dimensional orientation of whole cell membranes was demonstrated in whole cell multilayers using doxyl stearate spin label in A. vinelandii and the bacteriochlorophyll a dimer triplet signal, (BCHl a)T2, from the intracytoplasmic membrane-bound photosynthetic apparatus of R. rubrum. Subsequent analysis of the low-field signals, g = 4.3 and g = 3.6, of molybdenum-iron protein in whole cell multilayers of each organism showed orientation-dependent characteristics, although the properties of each were different. Specifically, as the normal to the membrane plane was rotated from perpendicular to parallel with the ESR magnetic field, the amplitude of the g = 3.6 signal decreased from maximum to about 37% of maximum in A. vinelandii and from maximum to about 88% of maximum in R. rubrum. The angular dependence of the g = 4.3 peak during rotation varied in A. vinelandii, but decreased from maximum to about 63% of maximum in R. rubrum. These data suggest that the molybdenum-iron protein of nitrogenase was oriented in response to the physical orientation of cellular membranes and that a structural association may exist between this nitrogenase component and membrane in these organisms. PMID- 2981551 TI - The effect of serum deprivation on the initiation of protein synthesis in mouse neuroblastoma cells. AB - Growth of mouse neuroblastoma cells becomes stationary when cultured in serum free medium. Within 60 h, the protein-synthesizing capacity of the cells declines to 25% as compared to that of exponentially growing cells. The transitional activity of the crude ribosomal salt washes from serum-deprived and control cells was compared in in vitro protein-synthesizing pH 5 systems. It appears that the ribosomal salt wash from serum-deprived cells has significantly (70%) lost its ability to support the translation of neuroblastoma poly(A)+ RNA. This activity of the ribosomal wash from serum-deprived cells can be restored to control level with rabbit reticulocyte initiation factor eIF-4B only. The ability of the ribosomal wash from serum-deprived cells to support the translation of encephalomyocarditis virus (EMC) and Semliki Forest virus (SFV) 42 S mRNA was tested. We found that EMC-mRNA is efficiently translated with the ribosomal salt wash from serum-deprived cells, whereas on the other hand the translation of SFV 42 S mRNA is severely impaired. Therefore, we conclude that in serum-deprived neuroblastoma cells protein synthesis is regulated in both a quantitative and a qualitative way. Modulation of the activity of initiation factor of protein synthesis eIF-4B is at least partly responsible for the observed (selective) blockade of protein synthesis in serum-deprived cells. PMID- 2981552 TI - Studies on mitochondrial type I topoisomerase and on its function. AB - We have reported previously that rat liver mitochondria contain a topoisomerase and have shown it to be distinct from the nuclear enzyme by its sensitivity to Berenil and ethidium bromide. We report here some additional characterization. The enzyme differs further from its nuclear counterpart in its failure to bind to ssDNA cellulose and its chromatographic behavior on Sephadex; the latter procedure yields an Mr of 44 000 for the mitochondrial and 70 000 for the nuclear enzyme. The topoisomerase is strongly associated with mitochondrial membranes; only 10% of the activity could be extracted. The pH optimum of the enzyme falls between 6.0 and 8.5, with an NaCl optimum of 0.13 M in 0.1 M Tris (pH 8.3). Dithiothreitol is required, while N-ethylmaleimide is inhibitory. Tosylphenylalanine chloromethyl ketone, a serine proteinase inhibitor, abolishes activity; another, phenylmethanesulfonyl fluoride, has no effect. Berenil, a non intercalating drug, and four of its analogues all inhibit with up to 100-fold differences in potency. No dependence on ATP, Mg2+, or both together could be shown. Neither novobiocin nor oxolinic acid shows any inhibitory effect. Nicked circles are generated in the presence of DMSO. These three observations are consistent with the topoisomerase being of the Type I class. Positively supercoiled pBR322 DNA, whose 6-8 positive turns were generated by altering solution conditions, is relaxed by the enzyme, indicating a lack of requirement for a negatively supercoiled substrate. We have also examined a partially purified preparation of the corresponding mitochondrial enzyme from mouse L cells. This enzyme is largely similar in properties to the rat liver enzyme. In isolated mitochondria, Berenil causes biphasic alterations in [3H]dATP incorporation into DNA, 10(-4) mM stimulating 2-fold, while higher concentrations inhibit. [3H]UTP incorporation into mitochondrial RNA also follows this pattern. PMID- 2981553 TI - Physical properties of a plasmid-like DNA from Euglena gracilis. AB - A small circular extrachromosomal DNA of the flagellate protozoan Euglena gracilis has been characterized as having a contour length of 11.3 kb, with a consistent restriction map. The buoyant density (rho = 1.717) and melting temperature (tm = 89 degrees C) both indicate a base content of 59% G + C. The DNA is found in both wild-type cells and those lacking plastids. The copy number is estimated to be about 1000. PMID- 2981554 TI - Effects of acidic and basic macromolecules on the activity of protein phosphatase 1. AB - The dephosphorylation of phosphorylase a by the catalytic subunit of protein phosphatase-1 obtained from rabbit skeletal muscle is inhibited by heparin in a noncompetitive manner with respect to phosphorylase a (Ki = 8 micrograms/ml). The inhibitory effect of heparin is also observed in the presence of effectors (e.g., glucose and AMP) modifying the dephosphorylation of phosphorylase a. Heat-stable protein inhibitors of protein phosphatase-1 can develop their inhibitory effect of the activity of protein phosphatase-1 even in the presence of heparin. The inhibitory effect of heparin and the heat-stable inhibitor-2 of phosphatase is additive. Polybrene, a heparin antagonist, prevented phosphatase-1 from the inhibition caused by heparin or the inhibitors. Proteins with basic character, histone fractions (H1, H3) and protamine sulfate, can counteract with the inhibitory effect of heparin, but they cannot intercept the actions of inhibitor 1 or -2. PMID- 2981555 TI - Possible role for metallothionein in protection against radiation-induced oxidative stress. Kinetics and mechanism of its reaction with superoxide and hydroxyl radicals. AB - Rabbit liver metallothionein-1 (Mr 6500), which contains zinc and/or cadmium ions, appears to scavenge free hydroxyl (.OH) and superoxide (O-.2) radicals produced by the xanthine/xanthine oxidase reaction much more effectively than bovine serum albumin (Mr 65 000) which was used as a control. Kinetic competition studies between metallothionein and either a spin trap for .OH or ferricytochrome c for O-.2 radicals, gave bimolecular rate constants of the order of kOH/MT approximately equal to 10(12) M-1 X s-1 and kO-2/MT approximately equal to 5 X 10(5) M-1 X s-1, respectively. The former value suggests that all 20 cysteine sulfur atoms are involved in this quenching process and that they all act in the diffusion control limit. The aerobic radiolysis of an aqueous solution of metallothionein, generating O-.2 and .OH radicals, induced metal ion loss and thiolate oxidation. These effects could be reversed by incubation of the irradiated protein with reduced glutathione and the appropriate bivalent metal ion. Metallothionein appears to be an extraordinarily efficient .OH radical scavenger even when compared to proteins 10-50-times its molecular weight. Moreover, hydroxyl radical damage to metallothionein appears to occur at the metal-thiolate clusters, which may be repaired in the cell by reduced glutathione. Metallothionein has the characteristics of a sacrificial but renewable cellular target for .OH-mediated cellular damage. PMID- 2981556 TI - Specific substrate for histone kinase II: a synthetic nonapeptide. AB - Based on the previously determined intrinsic substrate specificity of histone kinase II, a nonapeptide was synthesized which was a specific substrate for this enzyme. The Vmax value of phosphorylation of the peptide (Ala-Ala-Ala-Ser-Phe-Lys Ala-Lys-Lys-amide) was about the same as that for H1 histone and the apparent Km for the phosphorylation of the peptide was 0.2 mM, an order of magnitude higher than that for H1 histone. H1 histone inhibited the phosphorylation of the peptide, while the peptide did not inhibit the phosphorylation of H1 histone. In the crude extracts of calf thymus, spleen and liver, histone kinase II was the only enzyme which phosphorylated the synthetic peptide. The rate of phosphorylation of this peptide was used to determine the activity of histone kinase II in the crude extracts of several tissues obtained from different species. PMID- 2981557 TI - A kinetic study of the alkaline transitions in cytochrome c peroxidase. AB - Cytochrome c peroxidase undergoes a complex series of transitions between pH 8 and 14. Seven distinct spectral transitions occur between 4 ms and 24 h after exposure to alkaline pH. The fastest transition occurs within the mixing time of a stopped-flow instrument and converts the native high-spin ferric form of the enzyme to a low-spin form which may be the hydroxy complex of the enzyme. An apparent pKa of 9.7 +/- 0.2 relates the native and initial alkaline form of the enzyme. Three other low-spin enzyme forms are evident from the experimental data prior to denaturation of the enzyme and complete exposure of the heme to the solvent. The final denaturation process occurs with an apparent pKa of 10.3 +/- 0.3. PMID- 2981558 TI - Physicochemical characterization of the alkaline denaturation of cytochrome c peroxidase. AB - The alkaline denaturation of cytochrome c peroxidase and apocytochrome c peroxidase was investigated by analytical ultracentrifugation, gel-filtration chromatography, and circular dichroism. The results indicate that both cytochrome c peroxidase and the apoenzyme undergo extensive structural modifications upon exposure to alkaline pH, including dimer formation. The midpoint of the transition for dimer formation in the native enzyme occurs at pH 9.6 +/- 0.1, while loss of tertiary and secondary structure occurs with transition midpoints at pH 10.3 +/- 0.1 and pH 11.3 +/- 0.1, respectively. Studies performed in the presence of dithiothreitol and with carboxymethylated cytochrome c peroxidase indicate that dimer formation occurs via a disulfide crosslink involving the single cysteine residue in the enzyme. Denaturation of cytochrome c peroxidase in the presence of guanidine hydrochloride gave results similar to those obtained for the alkaline denaturation. PMID- 2981559 TI - Evidence for the existence of multiple forms of choline (ethanolamine) kinase in rat tissues. AB - In order to characterize the form of choline kinase in rat tissues, both electrophoretic and gel chromatographic patterns of choline kinase activity were compared in the liver, kidney, lung, whole intestine and carbon tetrachloride induced liver cytosols. Kinetic parameters of the reaction were also compared for the main forms of choline kinase protein from these tissues. The overall results suggested strongly that choline kinase does not exist in one particular active form but exists in multiple forms in rat tissues. In the study present here, the electrophoretic patterns of both choline kinase and ethanolamine kinase activities were compared in rat liver, kidney, lung and intestinal cytosols. The results strongly supported the view that both kinase activities are represented on the same enzyme protein(s) in each of the rat tissues examined. PMID- 2981560 TI - Binding and degradation of human high-density lipoproteins by human hepatoma cell line HepG2. AB - The catabolism of human HDL was studied in human hepatoma cell line HepG2. The binding of 125I-labeled HDL at 4 degrees C was time-dependent and reached completion within 2 h. The observed rates of binding of 125I-labeled HDL at 4 degrees C and uptake and degradation at 37 degrees C indicated the presence of both high-affinity and low-affinity binding sites for this lipoprotein density class. The specific binding of 125I-labeled HDL accounted for 55% of the total binding capacity. The lysosomal degradation of 125I-labeled HDL was inhibited 25 and 60% by chloroquine at 50 and 100 microM, respectively. Depolymerization of microtubules by colchicine (1 microM) inhibited the degradation of 125I-labeled HDL by 36%. Incubation of cells with HDL caused no significant change in the cellular cholesterol content or in the de novo sterol synthesis and cholesterol esterification. Binding and degradation of 125I-labeled HDL was not affected by prior incubation of cells with HDL. When added at the same protein concentration, unlabeled VLDL, LDL and HDL had similar inhibitory effects on the degradation of 125I-labeled HDL, irrespective of a short or prolonged incubation time. Reductive methylation of unlabeled HDL had no significant effect on its capacity to inhibit the 125I-labeled HDL degradation. The competition study indicated no correlation between the concentrations of apolipoproteins A-I, A-II, B, C-II, C-III, E and F in VLDL, LDL and HDL and the inhibitory effect of these lipoprotein density classes on the degradation of 125I-labeled HDL. There was, however, some association between the inhibitory effect and the levels of apolipoprotein D and C-I. PMID- 2981561 TI - The metabolism of leukotrienes in blood plasma studied by high-performance liquid chromatography. AB - The metabolism of leukotrienes (B4, C4, D4, and E4) within human plasma was studied and a simple sample preparation is presented. It was demonstrated that leukotriene E4 and leukotriene B4 were stable during incubation at 37 degrees C using the in vitro system. In contrast, leukotriene C4 was metabolized by gamma glutamyl transpeptidase activities into leukotriene D4 which was further metabolized by dipeptidase activities of plasma into leukotriene E4. The transition state inhibitor of gamma-glutamyl transpeptidase L-serine-borate decreased the metabolism of leukotriene C4 in plasma. Dilution of plasma demonstrated that the dipeptidase was more active compared to the gamma-glutamyl transpeptidase. The metabolizing activities of plasma were functionally characterized by fractionating the plasma proteins. PMID- 2981563 TI - Regulation and location of phosphatidylglycerol and phosphatidylinositol synthesis in type II cells isolated from fetal rat lung. AB - myo-Inositol decreases the synthesis of phosphatidylglycerol by type II cells isolated from fetal rat lung. Inositol addition also increases the synthesized amount of surfactant phosphatidylinositol. These observations indicate that at least part of the decreasing effect of inositol on phosphatidylglycerol formation is the result of competition between phosphatidylglycerol and phosphatidylinositol synthesis for a common pool of CDP diacylglycerol. Studies on the subcellular localization of enzymes measured under optimal conditions suggested that the enzymic activity required for the formation of phosphatidylglycerol is located mainly in the mitochondria, but most likely also for a small part in the endoplasmic reticulum, while the enzymic activity required for phosphatidylinositol formation is located in the endoplasmic reticulum. Inositol was found to inhibit glycerolphosphate phosphatidyltransferase in the microsomal fraction but not in the mitochondrial fraction derived from the type II cells, indicating that the competition between phosphatidylglycerol and phosphatidylinositol synthesis for CDP diacylglycerol takes place in the endoplasmic reticulum. This latter observation together with the observation of a switch-over from surfactant phosphatidylinositol to phosphatidylglycerol production around term indicate that the endoplasmic reticulum is the intracellular site of surfactant phosphatidylglycerol production. PMID- 2981562 TI - Diethylaminoethoxyhexestrol causes hypertriglyceridemia in guinea pigs. AB - Treatment of rats, monkeys and man with diethylaminoethoxyhexestrol causes phospholipid storage in liver and other tissues. However, this drug has not been reported to alter plasma lipoprotein levels. When guinea pigs were treated with diethylaminoethoxyhexestrol, the fasting plasma triacylglycerol levels increased dramatically, from 43 to 1281 mg/dl, after only five doses of 12.5 mg/kg. Diethylaminoethoxyhexestrol-treated guinea pigs had reduced postheparin lipase activity. In addition, in vitro assays showed that this agent inhibited guinea pig postheparin lipoprotein lipase. It is hypothesized that diethylaminoethoxyhexestrol causes hypertriglyceridemia in guinea pigs because these animals are known to have low levels of serum activator for lipoprotein lipase and may be unusually susceptible to agents that inhibit lipoprotein lipase activity. The ability to produce hypertriglyceridemia in guinea pigs provides an animal model in which the metabolic consequences of hypertriglyceridemia can be studied. PMID- 2981564 TI - Diacylglycerol lipase and kinase activities in rat brain microvessels. AB - Diacylglycerols can accumulate transiently in intact cells as a consequence of the degradation of phosphatidylinositol by phospholipase C, but little information is available concerning their metabolic fate in the vascular endothelium. Diacylglycerol lipase and kinase activities were measured in rat brain microvessel preparations. Lipase activity, measured by the release of free fatty acids, was much greater at pH 4.5 than at pH 7. The acid lipase was predominantly particulate and likely originated in lysosomes, whereas the neutral lipase was mainly soluble. The fatty acid at the sn-1 position of the diacylglycerol substrate was hydrolyzed faster than that at the sn-2 position at both pH 4.5 and 7. The 2-monoacylglycerol accumulated at pH 4.5 but not at 7 due to the presence of a monoacylglycerol lipase activity with a neutral pH optimum. The formation of phosphatidic acid (kinase activity) was also measured in microvessels. When lipase and kinase activities were measured simultaneously, the formation of phosphatidic acid from a 1-palmitoyl-2-[1-14C]oleoyl-sn-glycerol substrate was 4-fold greater than the release of fatty acid (oleate) from the sn 2 position. Introduction of arachidonic acid to the sn-2 position of the diacylglycerol substrate increased kinase activity but reduced lipase activity. The release of fatty acids from the sn-2 position of phosphatidic acid could not be detected. PMID- 2981565 TI - Intensity and kinetics of the respiratory burst of human neutrophils in relation to receptor occupancy and rate of occupation by formylmethionylleucylphenylalanine. AB - Studies on the relationship between the binding of fMet-Leu-Phe and the respiratory response in human neutrophils have been carried out under two different conditions of stimulus presentation, i.e., instantaneously and over a period of time. The main findings are as follows (1) Under the first condition the activation of the respiratory response reaches the maximum value very quickly, when the receptor occupancy is less than 20% that at equilibrium. After reaching this maximal value, the activated respiration progressively decreases, while the specific binding of the stimulant continues until equilibrium. (2) Under the second condition, i.e., when the stimulus to neutrophils is presented over a time of 1, 2 or 4 min, the respiratory response (and also the secretory one) is depressed or absent, and the initial rate of the binding (initial Vass) is lower, but the maximal values of the receptor occupancy at equilibrium and of the rate of receptor occupation (maximal Vass) are similar and only slightly lower than those reached under the condition of instantaneous presentation of the stimulus. (3) This form of desensitization is specific for fMet-Leu-Phe and does not consist of the inactivation of the target (NADPH oxidase), since neutrophils desensitized by the slow presentation of the peptide are able to respond to a second challenge with other stimulants. These results indicate that: (1) the efficacy of the stimulus-receptor complexes is short-lived; (2) the intensity of the respiratory response is dependent on the rate of reaching a threshold of binding; (3) when this initial rate is slow, owing to the slow presentation of the stimulus, a specific desensitization takes place, indicating the existence of a molecular mechanism, linked in some way to the initial rate of binding, that modulates the capacity of the stimulus-receptor complexes to transduce signals for cell responses. The physiological role of this type of desensitization is discussed. PMID- 2981566 TI - Mechanism of desensitization of neutrophil response to N formylmethionylleucylphenylalanine by slow rate of receptor occupancy. Studies on changes in Ca2+ concentration and phosphatidylinositol turnover. AB - Previous studies on the regulation of responses of neutrophils to fMet-Leu-Phe have demonstrated the relevance of the role of the rate of occupation of the receptors by the stimulant. When this rate is decreased by presenting the peptide to neutrophils over a period of time by means of an infusion pump, the activation of the respiratory burst and of the secretion is greatly depressed or is absent. This paper deals with further investigations on the mechanisms of this desensitization, which previous results have shown to consist of an uncoupling between the ligand-receptor complexes and the target for cell responses, caused by the deceleration of the initial rate of occupation of the receptors. The data presented here demonstrate that this desensitization is not linked to the formation of a negative intermediate such as cAMP, but is associated with: (i) a depression of the rate and magnitude of the phosphatidylinositol response (activation of phosphatidylinositol turnover measured as modification of incorporation of [32P]Pi and [3H]glycerol into phosphatidylinositol and phosphatidic acid); (ii) a deceleration of the rate of the release of bound Ca2+, without a decrease in the total quantity of Ca2+ liberated (measured as fluorescence changes of chlorotetracycline treated neutrophils); (iii) a slower rise of cytosolic free Ca2+ concentration [Ca2+]i, without a decrease in the magnitude of the final increase of [Ca2+]i (monitored with Quin 2). These findings, which are discussed in relation to the recent hypotheses on the transduction reactions of receptor-mediated stimuli for neutrophil responses, are consistent with a mechanism of desensitization involving decreased production of diacylglycerol by the hydrolysis of phosphatidylinositol and deficient activation of Ca2+-phospholipid-dependent protein kinase C. PMID- 2981567 TI - Induction of osteoblastic cell differentiation by forskolin. Stimulation of cyclic AMP production and alkaline phosphatase activity. AB - The effects of forskolin on differentiation of osteoblastic cells (clone MC3T3 E1) cultured in alpha-minimum essential medium containing 0.1% bovine serum albumin were investigated by assays of intracellular cyclic AMP level and alkaline phosphatase activity in the cells. Forskolin increased cyclic AMP production in the cells in a dose-related manner, the maximum increase being 250 fold above that of the controls. Alkaline phosphatase activity in the cells was also elevated as early as 24 h and rose to nearly its maximum at 48 h. The elevation was dose-dependent, with a maximum increase at 5 X 10(-6) M forskolin. Forskolin and prostaglandin E2 showed a supraadditive effect on cyclic AMP production in the cells and had an additive effect on alkaline phosphatase activity, whereas forskolin and dibutyryl cyclic AMP had little additive effect on either cyclic AMP production or enzyme activity. These results suggest that cyclic AMP is closely linked to the differentiation of osteoblastic cells in vivo. PMID- 2981568 TI - Prolactin uptake into liver endocytic components. Reduced sensitivity to chloroquine. AB - Subfractionation of hepatic Golgi fractions on Percoll gradients revealed two populations of lactogen-receptor enriched components with one pool of rho = 1.040 1.050 and a high-density component of rho = 1.053-1.064. 125I-labelled prolactin uptake into Percoll gradient subfractions demonstrated rapid accumulation into low-density elements and slower accumulation in high-density structures. Electron microscope radioautography demonstrated that silver grains were largely associated with lipoprotein-filled structures. Radiolabel was highly concentrated in these components especially the very pure higher-density component where enrichment over the homogenate was 184-fold at the peak time after injection (10 min). Administered chloroquine accumulated in the higher-density component but not in low-density elements, suggesting that the former are at an acid intraluminal pH. In contrast to the marked effect of chloroquine on insulin uptake into endocytic structures found in Golgi fractions (Posner, B.I., Patel, B.A., Khan, M.N. and Bergeron, J.J.M. (1982) J. Biol. Chem. 257, 5789-5799), little effect of this pH disrupting agent was observed on prolactin uptake. The difference between the effect of chloroquine on 125I-labelled prolactin and 125I labelled insulin uptake may reflect the greater stability of prolactin-receptor complexes in a low-pH environment. PMID- 2981569 TI - Soluble sphingomyelinase from human urine as antigen for obtaining anti sphingomyelinase antibodies. AB - A soluble form of lysosomal sphingomyelinase was partially purified from human urine using concanavalin A-Sepharose 4B, Sephadex G-100 and octyl-Sepharose 4B chromatography. The octyl-Sepharose 4B eluate was used to immunise a rabbit. The antiserum obtained was able to precipitate about 70% of the sphingomyelinase activity present in urine from control subjects. Both the immunoprecipitable and non-precipitable activities were found to be deficient in urine from patients with Niemann-Pick disease Type A and Type B. In contrast, both activities were present in urine from patients with Niemann-Pick disease Type C. The antiserum was able to precipitate about 80% of the sphingomyelinase activity present in an aqueous extract of placenta. PMID- 2981571 TI - Studies on structure-function relationships of human choriogonadotropins with C terminally shortened alpha-subunits. I. Receptor binding and immunologic properties. AB - Recombination products composed of the native beta-subunit and an alpha-subunit with an enzymatically shortened C-terminal region showed a diminished (less than 5 amino acids removed) or - in the case of des-(88-92)-alpha/native beta - a completely abolished ability to bind to testicular LH/hCG receptors of the rat. An antigenic determinant which is present in native hCG but not in the isolated subunits was not or incompletely expressed in the modified hormone species. Antigenic determinants which are characteristic for the isolated alpha-subunit, however, were not affected by removal of the C-terminal residues 88-92. The immunologic experiments indicate that hCG containing an alpha-subunit with a shortened C-terminal region differs from native hCG in its conformation. These conformational changes are probably responsible for the loss in receptor-binding ability. PMID- 2981570 TI - Modulation of collagen production in cultured fibroblasts by a low-frequency, pulsed magnetic field. AB - Primary cultures of chicken tendon fibroblasts have been exposed for various periods to a low-frequency, pulsed magnetic field, and the effects on protein and collagen synthesis have been examined by radioisotopic incorporation. Total protein synthesis was increased in confluent cells treated with a pulsed magnetic field for the last 24 h of culture as well as in cells treated for a total of 6 days. However, in 6 day-treated cultures, collagen accumulation was specifically enhanced as compared to total protein, whereas after short-term exposure, collagen production was increased only to the same extent as total protein. Levels of cyclic AMP were significantly decreased after 6-day pulsed magnetic field treatment, probably as a consequence of diminished adenylate cyclase activity. Exposure to pulsed magnetic field had no effect on cell proliferation or collagen phenotype. These results indicate that a pulsed magnetic field can specifically increase production of collagen, the major differentiated function of fibroblasts, possibly by altering cyclic-AMP metabolism. PMID- 2981572 TI - Studies on structure-function relationships of human choriogonadotropins with C terminally shortened alpha-subunits. II. Stimulation of adenylate cyclase activity and depletion of cytochrome P-450 in the rat testis. AB - Human choriogonadotropin (hCG) analogues, containing the native beta-subunit and alpha-subunits enzymatically shortened by 2-3 amino acid residues, were used for studying the influence of hCG on the content of microsomal progesterone-binding cytochrome P-450 in rat testis. When 2-3 residues have been removed from the alpha-subunit, the ability of the hormone analogue to stimulate adenylate cyclase of isolated rat Leydig cells was diminished by 55%. When the hCG analogue containing a des-(88-92)-alpha chain was applied, the residual activity of the adenylate cyclase was negligible. 18 h after administration to rats in vivo, the hormone species containing des-(Lys-91-Ser-92)-alpha or des-(90-92)-alpha, respectively, were found to have induced a decrease in microsomal cytochrome P 450 content with an effectiveness corresponding to their ability of stimulating the adenylate cyclase in vitro. However, when assayed 48 h after application, the desensitization of the microsomal cytochrome P-450 system had persisted in case of the hCG species containing a des-(90-92)-alpha chain but not in case of hCG consisting of des-(Lys-91-Ser-92)-alpha and a native beta-subunit. From these results, it is concluded that short-term effects of hCG on the microsomal content of progesterone-binding cytochrome P-450 are mediated by the stimulation of adenylate cyclase. In contrast, the long-lasting action of hCG on this system seems not to be exclusively mediated by the increase in intracellular cAMP. PMID- 2981574 TI - [Potentials of enzyme tests and radioisotope hepatography in detecting early functional changes in the liver]. AB - The enzymic tests and radionuclide hepatography were used to study and compare liver function after rabbits were exposed to tetrachloromethane poisoning. The activity of serum enzymes of cholinesterase, alkaline phosphatase, aldolase and leucine aminopeptidase was determined. Hepatography was made with the use of 198Au-colloid with an activity 0.74 MBC. The enzymic tests were demonstrated to be more sensitive than radionuclide hepatography in detecting the earliest parenchymatous lesions in the liver. The data obtained correlate with the data of the pathohistological examinations, which demonstrated the presence of marked vacuole parenchymatous fatty dystrophy. The authors recommend that the enzymic tests should be used for detecting early hepatic lesions induced by tetrachloromethane. PMID- 2981573 TI - Independence with respect to Ca2+ changes of the neutrophil respiratory and secretory response to exogenous phospholipase C and possible involvement of diacylglycerol and protein kinase C. AB - Exogenous phospholipase C induces in human neutrophils the activation of a respiratory burst, measured as O2 consumption and O-2 production, and of secretion of specific granules, measured as release of vitamin B-12 binding protein. The secretory response is minimal and follows the onset of the respiratory response. Studies carried out using cells prelabeled with [3H]glycerol and 32P on the molecular mechanism of the stimulations demonstrate that the effects are dependent on the formation of diacylglycerol by hydrolysis of different classes of glycerophospholipids. They are, however, independent of the activation of a 'phosphoinositide turnover' as occurs in cells stimulated with fMet-Leu-Phe. Furthermore, the respiratory and secretory responses to exogenous phospholipase C are not associated with modifications of cytosolic Ca2+ concentration, measured with the Quin-2 method, and of the release of bound Ca2+, measured with the membrane probe, chlorotetracycline. Apart from a quantitative difference, mostly regarding the ratio of the intensity of the respiratory and secretory responses, the effects caused by exogenous phospholipase C are qualitatively similar to those induced by phorbol myristate acetate and are probably linked to an involvement of protein kinase C, activated by diacylglycerol liberated in the plasma membrane. PMID- 2981575 TI - [Na, K-ATPase activity of the microsomal fraction of the rat brain exposed to insulin]. AB - The effect of insulin on the activity of Na, K-ATPase was studied in rat brain microsomes. Addition of insulin to the incubation medium in a dose of 0.18 U/ml coupled with strophanthine did not change the enzyme activity. The raising of the hormone dose to 0.36 U/ml was accompanied by inhibition of the enzyme activity. The incubation duration (10 and 30 min) did not influence the Na-pump. Preincubation of brain microsomes with insulin for 5 min significantly activated Na, K-ATPase. It has been thus demonstrated that insulin is capable of influencing the activity of Na, K-ATPase of rat brain microsomes in vitro. The effect obtained depends both on the dose of the hormone introduced into the incubation medium and the experimental conditions. PMID- 2981576 TI - [Content of phospholipid, cerebroside and cerebroside sulfate in the central nervous system of mice with acute experimental viral demyelination]. AB - A study was made of the content of phospholipids, cerebrosides and cerebroside sulfates in the central nervous system of mice with experimental acute viral encephalomyelitis. No considerable changes in phospholipid content were revealed. A significant drop in the content of cerebrosides and cerebroside sulfates was defected in the CNS, being more pronounced in the spinal cord of sick animals. The reduction in the content of glycolipids can be explained by myelin disintegration and by the effect of viruses on the olygodendrocytes in which cerebrosides and cerebroside sulfates are synthesized. PMID- 2981577 TI - [ATP-dependent proton translocation across the synaptic vesicle membrane in the brain of rats]. AB - Changes in pH in rat brain synaptic vesicles (SV) were studied with the use of the fluorescent slightly basic dye acridine orange. The pH value in isolated SV was found to be acidic, which was confirmed by the ionophore sensitive accumulation of the dye. Addition of ATP provoked further acidification of the intravesicular medium. The acidification rate reached a maximum after dissipation of the existing H+ gradient seen during preincubation in the absence of ATP. The ATP-dependent acidification was eliminated by the protonophore carbonylcyanide m chlorophenylhydrazone, H4Cl or the detergent triton X-199 (0.025%). Valinomycin inhibited the ATP-dependent translocation of H+ whatever the incubation medium (with KCl or NaCl). Dicyclohexylcarbodiimide, a known inhibitor of proton ATPases (100 microM) as well as ethylmaleimide (100 microM) completely blocked H+ translocation whereas oligomycin, a specific blocker of mitochondrial H+-ATPase, and ouabain did not influence that process. ATP induced H+ translocation only in the presence of Mn2+ or Mg2+ but not in the presence of Ca2+. The translocation of H+ was not affected by the replacement of univalent cations (KCl, NaCl or Cl), however, it was prevented completely upon replacement of the penetrating anion Cl by the non-penetrating anion O2-4 or upon replacement of the salts by sucrose. It is concluded that the ATP-dependent translocation of H+ in SV is mediated via H+-ATPase which maintains the low pH value in SV. PMID- 2981578 TI - [Role of endogenous free iron in activating lipid peroxidation in ischemia]. AB - Ischemia was simulated in rat liver perfused by physiological solution. The concentration of free iron and lipid peroxidation (LPO) products was measured 1, 2, 3, 4 and 5 hours after ischemia onset. The ESR method was used to measure free iron concentration. The LPO intensity was evaluated by the TBA test and by optical density at 232 nm. The content of free iron in cytoplasm increased in the course of ischemia with an increase in the concentration of LPO products. The content of free iron in the membranes remained unchanged. It is supposed that activation of LPO in ischemia may be caused by the appearance in the cytoplasm of a large amount of free iron. This iron can be liberated from ferritin in conditions of low oxygen concentration. PMID- 2981579 TI - [Effect of corticotropin on the dehydrogenase activity of cells and tissues]. AB - The authors studied the effect of native ACTH on dehydrogenase activity of isolated strips of rat diaphragm and suspension of E. coli cells, serotype O III:B4, grown on beef extract agar in a medium with different dehydrogenation substrates. ACTH activated dehydrogenase of rat diaphragm in a medium containing pyruvate, alpha-ketoglutarate, malate, beta-hydroxybutyrate, D-aspartic acid and did not alter it in a medium containing succinate. In contradistinction to rat diaphragm, ACTH activated dehydrogenase of E. coli cells whatever the substrates used (oxaloacetate, isocitrate, alpha-ketoglutarate, succinate, fumarate, malate, pyruvate, lactate, beta-hydroxybutyrate, glucose, D-aspartic acid. Synacthen (ACTH1-24) exerted a similar effect. It is suggested that the effects of ACTH are mediated via its influence on adenylate cyclase in the absence of receptors. PMID- 2981580 TI - [Effect of beta-neo-endorphin--an agonist of chi-opiate receptors on the neuronal activity of the cerebral cortex]. AB - Experiments on cats were made to examine the neuronal responses (the first zone of the somatosensory cortex and the 5th region of the parietal associative cortex) to microapplications of beta-neo-endorphine (an agonist of kappa-opiate receptors), morphine (an agonist of mu-opiate receptors), leuenkephalin (an agonist of delta-opiate receptors) and beta-endorphine (an agonist of mu, delta, epsilon-opiate receptors). beta-Neo-endorphine and other opioid peptides produced similar depressive changes in spontaneous activity. Naloxone in doses which block the depressive reactions of morphine, leu-enkephalin and beta-endorphine did not remove the depressive reactions of beta-neo-endorphine. Opioid peptides and morphine produced different changes in nociceptive-stimulated neuronal activity in the first zone of the somatosensory cortex and photostimulated neuronal activity in the 5th region of the parietal cortex. It is assumed that the different types of opiate receptors and their endogenous ligands (opioid peptides) play different functional roles in the preparation of the nociceptive and visual information in the cortex. PMID- 2981581 TI - [Characteristics of electrical activity in different sections of nerve endings in the frog]. AB - In experiments on frog sartorius neuromuscular preparations, the evoked electrical responses of nerve endings were recorded by extracellular microelectrodes. It was shown that in proximal parts of the nerve ending, the three-phase response (+ - +) with a high amplitude negative phase occurred due to motor nerve stimulation. With movement of the extracellular electrode in distal direction a certain increase of the initial positive phase and a significant decrease of the negative one were observed. At the end of the terminal that response transformed to the monophasic one (+). On local iontophoretic application of tetrodotoxin (TTX) to the recording site two components of the nerve ending response were revealed: TTX-insensitive and TTX-sensitive. A significant decrease of the TTX-sensitive component occurred along the course of the nerve ending. That component was absent from the distal synaptic areas. It is concluded that in frog nerve ending, the action potential propogates with decrement while depolarization of the end parts of the terminal is passive in nature. PMID- 2981582 TI - [Formation of superoxide radicals by the nuclear membranes of human brain tumors]. AB - It has been discovered that nuclear membranes isolated from human brain tumors are capable of cyanide-sensitive oxidation of adrenaline to adrenochrome in the presence of both NAD X H and NADP X H while the process of oxidation is inhibited by superoxide dismutase. At the same time nuclear membranes isolated from the brain tissue do not oxidize adrenaline to adrenochrome in the presence of both substrates. A possible relationship between the properties of the nuclear membranes and the degree of tumor malignancy is discussed. PMID- 2981583 TI - The family of human T-lymphotropic leukemia viruses: HTLV-I as the cause of adult T cell leukemia and HTLV-III as the cause of acquired immunodeficiency syndrome. PMID- 2981584 TI - Monoclonal antibody AHN-1 inhibits phagocytosis by human neutrophils. AB - The granulocyte-specific monoclonal antibody, AHN-1, immunoprecipitates two major surface-iodinated proteins of 105,000 and 145,000 to 150,000 daltons from normal human neutrophils. In this study, the effect of AHN-1 on a number of neutrophil functions was evaluated in vitro. Both complement- and antibody-mediated phagocytosis were inhibited when human neutrophils were pretreated with AHN-1 and opsonized bacteria were used as targets. The inhibition of phagocytosis was specific, in that lysosomal enzyme release and chemotaxis were not altered by treatment with AHN-1. AHN-1 did inhibit superoxide production by neutrophils in response to particulate stimuli, but not in response to the soluble stimulus, 12 O-tetradecanoylphorbol-13-acetate. The data indicate that one or both of these surface proteins may be important in the process of phagocytosis. AHN-1 should be useful in isolating and further characterizing the nature of these molecules. PMID- 2981585 TI - Cleavage of human von Willebrand factor by platelet calcium-activated protease. AB - In human platelet lysates prepared by addition of nonionic detergent (Triton X 100) or by sonication, the multimer composition and electrophoretic mobility of platelet von Willebrand factor (vWF) were consistently modified under conditions that would favor activation of the endogenous calcium-activated, sulfhydryl dependent neutral protease (CAP). By sodium dodecylsulfate-agarose gel electrophoresis, native platelet vWF contained some multimers that were larger than those characteristic of plasma vWF. Modified platelet vWF contained a multimer population equivalent to or smaller than that of plasma vWF plus an additional fast-migrating band. In crossed immunoelectrophoresis (CIE), modified platelet vWF was characterized by a more anodic distribution and the appearance of a distinct, cross-reactive, anodic component previously designated VIIIR:Ag fragment. In the presence of calcium, radiolabeled purified plasma vWF was also degraded by the protease in question, with a decrease in the apparent molecular weight of the reduced monomer from 230,000 to 205,000. The VIIR:Ag fragment isolated from the same degraded plasma vWF by preparative CIE was shown to be composed of an identical mol wt 205,000 subunit. Because cleavage of plasma or platelet vWF was inhibited by prior addition of leupeptin, EDTA, ethylene glycol bis (beta-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA), or N ethylmaleimide (agents known to inhibit platelet CAP) but was unaffected by numerous other protease inhibitors, including soybean trypsin inhibitor, benzamidine, hirudin, phenylmethylsulfonyl fluoride, aprotonin, or epsilon aminocaproic acid (none of which inhibits platelet CAP), we conclude that proteolysis of vWF observed in this study is a direct effect of CAP and is not mediated by way of secondary proteases. PMID- 2981586 TI - Neutrophils autoinactivate secretory products by myeloperoxidase-catalyzed oxidation. AB - The neutrophil response to inflammatory stimuli involves the formation of reactive oxygen species and secretion of granule enzymes. In studying secretion of vitamin B12 binding protein by human neutrophils, we noted a major decrease in total recoverable activity from the extracellular fluid plus the stimulated cells (54% of resting cells). Recovery of B12 binding protein from neutrophils exposed to phorbol myristate acetate or opsonized zymosan was significantly enhanced on addition of heme enzyme inhibitors (azide, cyanide) or catalase or when halide free medium was used. The changes in B12 binding protein recovery were attributable entirely to increases in extracellular fluid levels, and cell pellet content was unaffected. These data indicate extracellular destruction of functional B12 binding protein by the halide-dependent heme enzyme myeloperoxidase and H2O2. Kinetic studies demonstrated rapid secretion of B12 binding protein in the first two to five minutes, followed by its inactivation over the next 20 to 30 minutes. A cell-free extract of vitamin B12 binding protein was readily inactivated on exposure to purified myeloperoxidase, H2O2, and a halide. These findings document a functional interaction among products of the neutrophil specific granules (B12 binding protein), azurophil granules (myeloperoxidase), and metabolic burst (H2O2). They provide an interesting model for the modulation of the inflammatory response by oxidation of secretory products of neutrophils. PMID- 2981587 TI - Fechtner syndrome--a variant of Alport's syndrome with leukocyte inclusions and macrothrombocytopenia. AB - This study reports a family comprising four generations in whom nephritis, deafness, congenital cataracts, macrothrombocytopenia, and leukocyte inclusions were observed in varying combinations in eight of 17 members. The family differs from others reported in that their hematologic abnormalities include not only macrothrombocytopenia, but also small, pale blue cytoplasmic inclusions in the neutrophils and eosinophils. Light microscopic appearance of the inclusions resembled that of toxic Dohle bodies and inclusions of May-Hegglin anomaly, but their ultrastructural appearance was unique. The inclusions consisted of clusters of ribosomes and small segments of rough endoplasmic reticulum (RER). They lacked the parallel 10-nm filaments characteristic of May-Hegglin anomaly and the parallel strands of RER seen in toxic Dohle bodies. Platelets were large, but their light and ultrastructural appearance was not significantly different from normal platelets. Platelet aggregation in response to epinephrine, arachidonate, thrombin, adenosine diphosphate, collagen, and ristocetin was normal. Levels of nucleotides and serotonin were elevated in proportion to cell volume. The concentration of adenosine triphosphate secreted and the percentage of arachidonic acid converted to thromboxane B2 were proportional to cell number. Deafness was high-tone sensorineural. Renal disease ranged from microscopic hematuria to end-stage renal failure necessitating dialysis and kidney transplantation. All affected adults had cataracts. This family represents a variant of Alport's syndrome with cataracts and leukocyte inclusions that, because of the associated macrothrombocytopenia, may be confused with May-Hegglin anomaly. PMID- 2981588 TI - Lymphokine abnormalities in aplastic anemia: implications for the mechanism of action of antithymocyte globulin. AB - Anti-thymocyte globulin (ATG) provides effective therapy for many patients with aplastic anemia, and its mechanism of action has been presumed to be secondary to lymphocytotoxicity. However, our studies of lymphocyte function in aplastic anemia show marked abnormalities of lymphokine production, which ATG may modulate. In 12 of 17 patients with aplastic anemia, interleukin 2 (IL2) production was markedly elevated in vitro (P less than .01 by paired statistical analysis). Expression of the IL2 receptor, or Tac antigen, on peripheral lymphocytes assessed by flow microfluorometry was also increased above the normal range in 11 of 15 cases. Studies of ATG suggested that it might act to stimulate lymphocyte function. In vitro, ATG is a mitogen, as measured by incorporation of 3H-thymidine into blood mononuclear cells; the response of cells to ATG from patients with aplastic anemia was exaggerated in comparison with normals. Cell proliferation was accompanied by production of IL2 to levels that were, in some cases, similar to those obtained with lectin stimulation. Finally, supernatants from lymphocytes cultured in the presence of ATG were able to replace adherent cells in providing growth factors for the support of nonadherent cells in methylcellulose hematopoietic colony assays. These results provide a mechanism for an "immunostimulatory" action of ATG in effecting hematopoietic response in some patients with aplastic anemia. PMID- 2981589 TI - Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophils by immunogold. AB - Colloidal gold was used as a marker for immunoelectron microscopy to localize lactoferrin (LF) and myeloperoxidase (MPO) in human peripheral blood neutrophils. Cells were reacted with monospecific antibodies against LF or MPO and then with gold-labeled antiglobulin. MPO cytochemistry was also associated with immunologic detection of LF. Immunologic labeling of thin sections after embedding in glycol methacrylate gave good ultrastructural morphology and specific localization of both proteins. MPO was detected in the large azurophil granules, whereas LF was consistently localized in the matrix of another population of morphologically distinct granules, smaller and more numerous than azurophil granules. When cytochemical detection of MPO was coupled with immunologic detection of LF, LF was observed in the population of MPO-negative granules, which were identified as specific. This was confirmed on cells that were permeabilized with saponin and stained for LF and MPO before embedding. No other neutrophil organelles displayed labeling for LF; other blood cells also were unreactive for LF. In the bone marrow, myeloblast and promyelocyte granulations were not stained and LF containing granules appeared at the myelocyte stage. In conclusion, we confirm previous biochemical and light microscopic studies by ultrastructural demonstration of LF and MPO in two categories of granules, the specific and azurophil granules, respectively. The method described in this article avoids disruption caused by cell fractionation procedures. In the future, other intragranular proteins can be localized by a similar approach. PMID- 2981590 TI - Intragenic Factor IX restriction site polymorphism in hemophilia B variants. AB - This study includes 47 normal subjects and 25 hemophilia B patients without inhibitor(s), showing different factor IX coagulant activity and antigen levels. Genomic DNA, digested with various restriction endonucleases, was hybridized with two different factor IX probes, ie, the cDNA and the subgenomic probe for the intragenic TaqI polymorphic site. cDNA restriction patterns suggest absence of gross rearrangements and/or deletions in all hemophilic patients. The frequency of the X chromosome bearing the TaqI polymorphic site is 0.32 +/- 0.09 in hemophilic subjects v 0.36 +/- 0.06 in normal control subjects, the latter value being comparable to that reported for the normal British population. No association between this polymorphism and hemophilia B variants has been observed, thus indicating that a wide spectrum of mutations underlies this blood clotting disorder and particularly each of its variants. PMID- 2981591 TI - Myeloperoxidase: its structure and expression during myeloid differentiation. AB - Myeloperoxidase (MPO) is a major protein present in myeloid cells and is used by these cells to help kill microbes. The human promyelocytic HL-60 line can be induced to differentiate to granulocytes or macrophagelike cells. Poly (A) containing RNA was isolated from HL-60 granulocytes, HL-60 macrophages, HL-60 blasts, and normal human granulocytes. The mRNA was translated in a reticulocyte lysate system in the presence of 35S-methionine. The MPO was precipitated from the lysate with rabbit IgG antiserum to human MPO. The resulting precipitate from HL-60 blasts gave a major band of radioactivity of approximately 77,000 daltons and another band at approximately 46,000 daltons on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The MPO identity of the labeled bands was confirmed by cold competition. The relative mRNA activity expressed as a percentage of radioactivity incorporated into MPO (77,000-dalton band) as compared with total trichloracetic acid (TCA) precipitable radioactivity was 0.2%. Negligible mRNA activity for MPO was present in HL-60 granulocytes, HL-60 macrophages, and normal human granulocytes. Pulse-chase experiments showed that MPO was an approximate 75,000-dalton major band and 77,000-dalton minor band of radioactivity after HL-60 blasts were labeled for 1/2 hour with 35S-methionine and the cell lysate immunoprecipitated and subjected to SDS-PAGE. The chase experiments (one to 24 hours) showed that the 77,000- and 75,000-dalton bands of radioactivity were replaced with two major bands (55,000 and 15,000 daltons) and one minor band (approximately 39,000 daltons) of radioactivity. Six-hour 35S methionine labeling experiments showed that the relative rate of MPO synthesis compared with total TCA precipitable radioactivity was 0.5% in HL-60 blasts and almost negligible in HL-60 macrophages and granulocytes, normal human granulocytes, and B-lymphocytes. The KG-1 myeloblasts and KG-1a early myeloblasts synthesized a small amount of the 75,000-dalton MPO protein. Although HL-60 cells no longer synthesized MPO after differentiation, HL-60 granulocytes and HL-60 macrophages continued to contain MPO as measured by enzyme activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981592 TI - Antibodies reactive with human T cell leukemia viruses in the serum of hemophiliacs receiving factor VIII concentrate. AB - The third member of the family of T cell leukemia viruses (HTLV III) has been proposed as the primary etiologic agent of the acquired immunodeficiency syndrome (AIDS). A high risk of AIDS has been reported among patients with hemophilia, particularly those with factor VIII deficiency who receive commercial clotting factor concentrates. In a prevalence survey conducted between September 1982 and April 1984, initial serum samples from 74% of hemophiliacs who had ever been treated with commercial factor VIII concentrate, 90% of those frequently treated with factor VIII concentrate, and 50% of those treated with both factor VIII and factor IX concentrates had antibodies reactive against antigens of HTLV III, compared with none of the hemophiliacs treated only with factor IX concentrate or volunteer donor plasma or cryoprecipitate. Two of the seropositive patients have developed AIDS-related illnesses, and a third patient died of bacterial pneumonia. One initially seronegative patient developed antibodies against HTLV III during the study and is currently well. The predominant antibody specificities appear directed against p24 and p41, the presumed core and envelope antigens of HTLV III, suggesting that factor VIII concentrate may transmit the p24 and p41 antigens of HTLV III. However, the presence of infectious retroviruses in clotting factor concentrates and the effectiveness of screening and viral neutralization procedures remain to be determined. PMID- 2981593 TI - IgG-antibodies to HTLV-III in patients with AIDS, LAS, and persons at risk of AIDS in West Germany. AB - In a first seroepidemiological study on the prevalence of the human T lymphotropic retrovirus HTLV-III in West Germany, sera of 26 patients with acquired immunodeficiency syndrome (AIDS), 33 patients with lymphadenopathy syndrome (LAS) or AIDS related complex (ARC), and 113 homosexual men at risk of AIDS were screened for IgG antibodies to HTLV-III by an enzyme linked immunosorbent assay (ELISA). 22 out of 26 AIDS-patients (84.6%), 24 out of 33 LAS patients (72.7%), and 44 out of 113 healthy homosexual men with increased risk of AIDS (38.9%) were found positive for antibodies to HTLV-III. Heterosexual controls including healthy laboratory workers and medical personnel with contact to AIDS patients did not show antibodies to HTLV-III. The HTLV-III antibodies analyzed predominantly recognize a protein of molecular weight 41,000 (p41). PMID- 2981594 TI - Fibronectin plasma levels after cadaver kidney transplantation. AB - The fibronectin plasma levels of 17 patients undergoing cadaver kidney transplantation were determined serially in the postoperative course using laser nephelometry. While 9 patients retained their grafts (group A), the grafts of 8 patients had to be removed (group B), mainly due to rejection, 3 patients had severe infections. In both groups a significant drop of the plasma fibronectin occurred after surgery. There were no significant differences between the two groups in the mean fibronectin levels. After the initial drop the group A patients exhibited rising values leading to a stable level. Progressively declining fibronectin plasma concentrations were found in 2 of 3 severely infected patients. Fluctuating values were found in 3 group B patients without a clear correlation to the rejection crises or the kidney function. The data suggest that the fibronectin plasma level does not seem to be a prognostic marker for graft rejection. But it might be useful in the important and often difficult differentiation between rejection and infection. PMID- 2981595 TI - Thalassaemia and Hodgkin lymphoma. PMID- 2981596 TI - The value of ultrasound in the diagnosis and treatment of small hepatocellular carcinoma. AB - Thirty-two patients with a solitary hepatocellular carcinoma of less than 4 cm diameter were treated during the 13 years from 1971 to 1983, two-thirds of them since 1981. Serum alpha-fetoprotein levels proved to be an excellent diagnostic tool for these early stage carcinomas which are generally asymptomatic. Ultrasonography forms an invaluable test for the detection of occult liver cancer and has enhanced the ability to detect small sized lesions. The accuracy of this method increased from 25 per cent for the period 1971-1980 to 89 per cent for the period 1981-1983. The cumulative survival rates for the 32 patients treated by hepatic resection at 1, 2 and 5 years were 85, 70 and 22.2 per cent (excluded from these figures are four operative deaths giving a 12.5 per cent operative mortality). PMID- 2981597 TI - A need for new converting enzyme inhibitors? PMID- 2981598 TI - Infection with parvovirus during pregnancy. PMID- 2981599 TI - Serial estimation of serum angiotensin converting enzyme activity during and after pregnancy in a woman with sarcoidosis. AB - Serum angiotensin converting enzyme activities were estimated during pregnancy and the puerperium in a woman with sarcoidosis and a series of normal women. In the patient with sarcoidosis angiotensin converting enzyme activity was raised during pregnancy, particularly at 21 weeks' gestation, yet she remained well with no symptoms to suggest relapse of sarcoidosis. Serum angiotensin converting enzyme activity may not be of value in monitoring sarcoidosis activity during pregnancy. PMID- 2981600 TI - Loss of form in young athletes due to viral infection. PMID- 2981601 TI - Serological response of melanoma patients to vaccines prepared from VSV lysates of autologous and allogeneic cultured melanoma cells. AB - Melanoma patients were vaccinated with cell-free lysates prepared from vesicular stomatitis virus (VSV)-infected cultured autologous and allogeneic melanoma cells. Eleven patients received vaccines produced from the melanoma cell line SK MEL-13. This cell line, derived from the melanoma of Patient AH, expresses a differentiation antigen (initially defined by autologous antibody) that is restricted to melanomas and other cells of neural crest origin (an example of a Class 2 melanoma antigen). Thirteen patients received vaccines prepared from autologous melanoma cells, the only known source of autologous unique (Class 1) melanoma antigens. VSV lysates were used for vaccination because VSV infection of tumor cells has been shown to augment the immunogenicity of tumor antigens. All patients but one vaccinated with VSV lysates of autologous melanoma cells developed antibodies against VSV, and all patients vaccinated with VSV lysates of SK-MEL-13 developed antibodies against HLA-related antigens. Antibodies against a Class 1 (unique) melanoma antigen were detected in only one case, and antibodies against Class 2 (shared) melanoma antigens were not found in any of the patients. The authors conclude that VSV lysates of melanoma cells are not effective in increasing the serologic response of melanoma patients to Class 1 or 2 melanoma antigens. PMID- 2981602 TI - Changes in ultrastructure and function of hypoxic V79 fibroblast cells after treatment with misonidazole. AB - Morphologic and enzymatic changes due to exposure to the radiosensitizing chemical, misonidazole, have been identified in V79 cells grown in a system in which oxygen tensions and culture density have been controlled. Misonidazole prevented the increase in mitochondrial size normally seen during exposure of these cells to conditions of moderate hypoxia (2 X 10(3) ppm O2). Mitochondrial size was also significantly decreased in cells from exponential cultures exposed to 1 mmol/l misonidazole. Morphologic changes to the mitochondria that varied from data reported elsewhere were also noted. Misonidazole caused a significant initial decrease in cytochrome oxidase activity after 4 hours of exposure of aerobic and moderately hypoxic cultures that did not return to normal in chronically hypoxic cells during continued exposure to the drug. PMID- 2981603 TI - Diagnostic and prognostic value of serum copper and plasma fibrinogen in hepatic carcinoma. AB - To investigate the diagnostic and prognostic value of several biochemical tests in primary liver tumors, the authors studied 36 cases (4 cholangiocarcinomas and 32 hepatocellular carcinomas, 10 of which were associated with cirrhosis) and 47 cases of liver cirrhosis, all with morphologically proven diagnosis. Serum copper (SCu) and plasma fibrinogen (PF) appeared the most useful tests in differential diagnosis between tumors and cirrhosis. In liver tumors, mean SCu level was 200.50, standard deviation (SD) 47.17 micrograms/dl (121.40, SD 25.90 micrograms/dl in cirrhosis; P less than 0.001). PF level was 461.78, SD 151.25 mg/dl in tumors (275.30 SD, 124.40 mg/dl in cirrhosis; P less than 0.001). SCu had a good sensitivity (0.80) and a high specificity (0.92) at a cutoff value of 160 micrograms/dl; when the cutoff level was raised to 170 micrograms/dl, the specificity increased to 1, with a sensitivity of 0.77. The combination of SCu and PF improved the diagnostic value slightly. Moreover, with an estimated frequency of tumor in cirrhosis of 10%, SCu had a positive predictive value of 1 (cutoff, 170 micrograms/dl) and a negative predictive value of 0.97. In nine patients SCu levels decreased after surgical removal of tumor; five other patients, sequentially studied, showed an increase of SCu level that correlated with the progression of the disease. Finally, patients with longer survival had a lower SCu level. These findings suggest that SCu level may be used as a screening test for early detection of neoplastic degeneration, and it is correlated with the extension of tumor mass. PMID- 2981604 TI - Morphologic study of three cases of insulinoma. Histochemical and ultrastructural studies. AB - Ultrastructural and immunocytochemical features of three human insulinomas, one with metastases and the other two with psammoma bodies and amyloid, respectively, are described. Neoplastic cells of all patients reacted with antihuman insulin antibody using the peroxidase-antiperoxidase (PAP) method. Ultrastructurally, neoplastic cells consisted of light cells and dark cells, both of which had a large number of secretory granules, microfilaments, ceroid bodies, and phagolysosomes. Psammoma bodies seemed to originate from neoplastic cell debris in one tumor. In another case, amyloid fibrils appeared to be produced by tumor cells, and there was a close association of amyloid with secretory granules. PMID- 2981605 TI - Radiation therapy for pineal and suprasellar germ cell tumors. AB - Radiation therapy (XRT) was used in the treatment of 25 patients with tumors of the pineal and suprasellar locations. A tissue diagnosis was obtained before XRT in 5 patients, and 20 were irradiated without histologic verification. The overall survival rate is 80% (76% with no evidence of disease [NED]). Megavoltage XRT was delivered to the entire neuraxis in 22 patients, and 86% (19/22) are alive from 4 to 88 months (median, 30 months) after treatment. In two of three patients treated only to local fields, tumor recurred in the spine; both are dead of disease. Biopsy-proven germinomas and multiple midline tumors responded favorably to XRT, whereas solitary pineal tumors and teratomas with marker positivity (human chorionic gonadotropin, alpha-fetoprotein) did not respond as well. The endocrinologic presentation, tumor marker status, and early response to radiation measured on computed tomography are useful means for selecting patients for radiation therapy. PMID- 2981606 TI - Karyotypic variation of cells from murine thymic lymphomas during short-term culture. AB - The present study examined the balance in the proportion of cells with either trisomy 15 or a diploid chromosome complement during short-term culture in vitro. Neonatal CFW/D mice were given an intrathymic injection of an endogenous virus (DMBA-LV) isolated from a DMBA-induced thymic lymphoma. The thymus was removed at the terminal leukemic stage, and the cells from each enlarged thymus were cultured in quintuplet. At 0, 6, 12, 24, and 48 hr, cultures were given colcemid for 2 hr prior to harvesting for chromosome examination. The analyzed lymphomas can be divided into three groups according to the characteristics of their stem lines and sidelines, as revealed by direct chromosome preparation (0 hr). Two groups consisted of lymphomas carrying a stem line with either trisomy 15 (group 1) or a diploid karyotype (group 2). A third group (group 3) consisted of lymphomas with a mixture of an abnormal stem line and sidelines. Chromosome analysis of cultures at subsequent times showed that there were fluctuations in the proportion of cells characterized as either stem line or sideline in all tumors at different times during culture. The initial sideline in group 1 tumors had a diploid complement. Although the proportion of the diploid cells increased in 5 of the 6 tumors following either 24 or 48 hr in culture, cells with a diploid karyotype remained as a sideline throughout the culture period. Similarly, the two tumors analyzed in group 2 showed that the initial sideline that contained trisomy 15 increased temporarily to become the stem line following 6 or 12 hr in culture. Group 3 tumors showed a slight variation in the proportion of cells with an abnormal stem line or sideline during culture. Considering all the data from the 10 tumors, 7 tumors had cells with a diploid karyotype, and, although the proportion of cells with this karyotype increased (4 tumors) or persisted (3 tumors) during 48 hr in culture, it remained as a sideline and was not overgrown by trisomy 15 stem line. PMID- 2981607 TI - Abnormalities of chromosomes 1 and 11 in Wilms' tumor. AB - Cytogenetic studies were performed successfully on 14 cases of Wilms' tumor. Six cases had detectable rearrangements of 11p, and 7 cases had structural abnormalities of chromosome #1. Two cases with advanced stage disease showed multiple translocations, resulting in hypodiploidy with a normal DNA complement. PMID- 2981608 TI - Cytogenetic analysis in a case of cancer of the male breast. AB - Sequential chromosome banding of direct preparations from an infiltrating ductal carcinoma of the left breast of a male, aged 56 years, showed a diploid chromosome range with a mode at 44. Consistent monosomy of chromosomes #2, #3, #4, #6, #11, #15, and #17 and nullisomy of chromosomes #1, #8, and #12 were found. In addition, each cell contained 11-14 markers and 1-7 abnormal chromosomes. Altogether, 16 markers were characterized, and 2 of these involved the long arm of chromosome #1. This chromosome pattern is similar to that in diploid breast carcinomas of the female. PMID- 2981609 TI - Localization of the oncogene c-Ha-ras1 outside the aniridia-Wilms' tumor associated deletion of chromosome 11(del 11p13) using somatic cell hybrids. AB - Hybrid cell lines, obtained after fusion of rodent cells with leukocytes from a patient with the aniridia-Wilms' tumor syndrome and carrying a specific constitutional deletion in chromosome #11 (del.11p13), were assayed for the presence of the c-Ha-ras1 oncogene. This sequence has recently been assigned to the p-arm of chromosome #11 and, hence, has been suggested to be involved in the development of renal tumors in patients with this syndrome. Positive hybridization of a cellular Ha-ras1 probe to hybrid cell DNA was observed, irrespective of whether the normal chromosome #11 or its deleted homologue was present. The results presented here suggest that c-Ha-ras1 is located outside the region 11p12-11p14, bounded by the chromosomal breakpoints observed in the patient used. Therefore, we conclude that predisposition of aniridia patients to develop Wilms' tumors is not due to a constitutional deletion of one of the c-Ha ras1 alleles. PMID- 2981610 TI - Role of active oxygen species in the photodestruction of microsomal cytochrome P 450 and associated monooxygenases by hematoporphyrin derivative in rats. AB - The cytochrome P-450 in hepatic microsomes prepared from rats pretreated with hematoporphyrin derivative was shown to be rapidly destroyed in the presence of long-wave ultraviolet light. The photocatalytic destruction of the heme-protein was dependent on both the dose of ultraviolet light and of hematoporphyrin derivative administered to the animals. The destructive reaction was accompanied by increased formation of cytochrome P-420, loss of microsomal heme content, and diminished catalytic activity of cytochrome P-450-dependent monooxygenases such as aryl hydrocarbon hydroxylase and 7-ethoxycoumarin O-deethylase. The specificity of the effect on cytochrome P-450 was confirmed by the observation that other heme-containing moieties such as myoglobin and cytochrome c were not susceptible to photocatalytic destruction. The destruction of cytochrome P-450 was a photodynamic process requiring oxygen since quenchers of singlet oxygen, including 2,5-dimethylfuran, histidine, and beta-carotene, each substantially diminished the reaction. Scavengers of superoxide anion such as superoxide dismutase and of H2O2 such as catalase did not protect against photodestruction of cytochrome P-450, whereas inhibitors of the hydroxyl radical, including benzoate, mannitol, and ethyl alcohol, did afford protection. These results indicate that lipid-rich microsomal membranes and the heme-protein cytochrome P 450 embedded therein are potential targets of injury in cells exposed to hematoporphyrin derivative photosensitization. PMID- 2981612 TI - Evidence for multiple steps in neoplastic transformation of normal and preneoplastic Syrian hamster embryo cells following transfection with Harvey murine sarcoma virus oncogene (v-Ha-ras). AB - Neoplastic development of Syrian hamster embryo (SHE) cells in culture is a multistep process in which intermediate or preneoplastic cells can be identified and isolated. In an attempt to further characterize normal and preneoplastic cells, we have compared their susceptibilities to neoplastic transformation following transfection with cloned DNA of the oncogenic virus, Harvey murine sarcoma virus (HaMSV). Normal SHE cells, which are stably nontumorigenic when injected in nude mice, are competent to take up and express exogenous DNA as demonstrated by transfection experiments with pSV2-neo DNA and certain viral DNAs. SHE cells treated with 5 micrograms of HaMSV DNA per dish remained nontumorigenic. Colonies of SHE cells, isolated after cotransfection with HaMSV and pSV2-neo DNA and selection for G418 antibiotic resistance, expressed Harvey murine sarcoma virus oncogene (v-Ha-ras) RNA and were initially morphologically altered; however, all colonies senesced when subcultured. In contrast, transfection of the cells with polyoma virus DNA alone or HaMSV DNA plus MC29 viral DNA (pSVv-myc) and then injection of the cells into nude mice resulted in progressively growing tumors of hamster origin within 3 to 5 weeks. A preneoplastic cell line, DES-4, isolated after treatment of SHE cells with the human carcinogen diethylstilbestrol, was chosen for comparative analyses. These immortalized cells are nontumorigenic and excellent recipients for exogenous DNA. In contrast to SHE cells, DES-4 cells were highly susceptible to neoplastic transformation following transfection with HaMSV DNA. To further investigate the role of HaMSV DNA in the neoplastic transformation of DES-4 cells and to determine whether this occurred as a single step, clones of DES-4 cells cotransfected with pSV2-neo and HaMSV DNAs were selected by antibiotic resistance and characterized. There was a good correlation between tumorigenicity and expression of v-Ha-ras DNA; however, the clones were highly variable in terms of their latency periods in vivo and anchorage-independent growth. Neither of these two parameters correlated with the level of expression of v-Ha-ras RNA. All of the cell lines derived from tumors and reinoculated into nude mice had short latency periods in vivo, were highly anchorage independent, and had high levels of v-Ha-ras expression. These results suggest that, in these experiments, v-Ha ras expression was necessary, but not sufficient, for the tumorigenicity of DES-4 cells and that additional changes in the cells were acquired.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981611 TI - Receptors for plasminogen activator, urokinase, in normal and Rous sarcoma virus transformed mouse fibroblasts. AB - We have prepared a conjugate of the plasminogen activator urokinase (UK) and ferritin, which maintains fibrinolytic activity. Monolayers of BALB/c-3T3 cells and of Rous sarcoma virus-transformed highly malignant line AA12-3T3, subcultured in plasminogen-free serum, were incubated with UK-ferritin at 0 degree and processed for transmission electron microscopy. Under these conditions, both of the lines showed specific receptors on the cell surface that were distributed in singlets, in small or large clusters. In the presence of excess native UK, the binding of ferritin was reduced by 99%, indicating the interaction of UK:ferritin with a specific receptor. The ligand-receptor interaction involves the catalytic site of UK, since the binding was completely impaired by preincubation of UK:ferritin with p-aminobenzamidine, a competitive inhibitor of the catalytic site of UK. The number and density of receptors decreased about one order of magnitude on the membrane of AA12 cells when compared with normal 3T3 cells. Saturation kinetics, using 125I-labeled UK, indicate the presence of 4 X 10(4) and 2.5 X 10(3) receptors on the membrane of 3T3 and AA12 cells, respectively. At 37 degrees, UK:ferritin redistributed on the plane of the membrane, in a process which was faster in malignant than in normal cells. Ferritin particles clustered in large groups on coated areas of the surface and were internalized by adsorptive pinocytosis. After 10 min at 37 degrees, the vesicles showed a progressively deeper internalization and a fusion with lysosomes, and some were observed in the Golgi complex area. Since the experiments were planned in order to exclude the presence of protease-nexin in the incubation medium, these data suggest the existence of a plasminogen-independent novel receptor for the catalytic site of plasminogen activators, the number on the cell surface of which decreases in Rous sarcoma virus-transformed mouse fibroblasts. PMID- 2981613 TI - Anti-transferrin receptor antibody linked to Pseudomonas exotoxin as a model immunotoxin in human ovarian carcinoma cell lines. AB - The present in vitro study was performed to evaluate the potential usefulness of immunotoxins in treating human ovarian carcinomas. A monoclonal antibody against the human transferrin receptor was covalently linked to Pseudomonas exotoxin. The activity of this immunotoxin (anti-TFR-PE) was studied in five ovarian carcinoma cell lines, a breast carcinoma cell line (MCF-7), and in KB cells. The ovarian carcinoma cell lines included one previously established cell line (A1847) and four recent isolates obtained from the malignant ascites of patients with metastatic ovarian carcinoma (OVCAR cell lines). While all cell lines showed inhibition of protein synthesis by anti-TFR-PE, there were quantitative differences when the level of protein synthesis was assayed after a 12-hr incubation with the immunotoxin. These differences resulted from different kinetics of anti-TFR-PE activity in the various cell lines. Higher levels of cellular binding and internalization of anti-TFR were shown to contribute to increased toxicity of anti-TFR-PE. Verapamil increased the rate of protein synthesis inhibition and thus enhanced the toxicity of anti-TFR-PE in the OVCAR cell lines. PMID- 2981614 TI - Biochemical analysis of human ovarian cancer-associated antigens defined by murine monoclonal antibodies. AB - Two monoclonal antibodies (MOv1 and MOv2) raised against a membrane preparation of a human surgical specimen from a mucinous ovarian cystoadenocarcinoma were used to biochemically define their target antigens. The heating of peritumoral mucus-soluble extracts and the sialidase treatment of crude membrane preparations did not affect the binding capacity of MOv1 and MOv2, which, on the contrary, was significantly reduced by periodate oxidation of the same materials. Pronase digestion completely solubilized MOv1-defined antigens, whereas MOv2-defined antigens were only partially solubilized. This, however, did not affect antibody binding with digested products. These data suggest that carbohydrate residues of recognized molecules constitute the antigenic determinants and that sialic acid residues are not involved. Gel filtration on Sepharose 4B of the peritumoral mucus, solubilized either by 200 mM NaCl or Pronase, revealed that most of the antigenic activity eluted in the void-volume fractions with a high carbohydrate content and in the included fractions before the elution volume of the ferritin standard protein. When CsCl gradient equilibrium ultracentrifugation of the solubilized mucus was used, MOv1-recognized antigens sedimented with a density of 1.45 g/ml, while the MOv2-defined epitope was carried by molecules with a density of 1.52 g/ml as well as by molecules with a lower density. Using thin-layer chromatography of organic solvent extracts obtained from mucus and crude membrane preparations, only MOv2-positive molecules could be resolved as a single band of glycolipid. Altogether, these data suggest that the antigens detected by MOv1 are mainly mucins whereas the determinant recognized by MOv2 is carried by both mucins and a glycolipid. To analyze the diagnostic potential of MOv1- and MOv2 recognized molecules, we tested their presence, as soluble products, in supernatants of tumor cell lines and in peritoneal effusions from cancer patients. To this aim, we developed an immunoradiometric assay using the same monoclonal antibody in insolubilized and soluble form. Whereas MOv1 immunoradiometric assay was always negative, by MOv2-immunoradiometric assay it was possible to detect the relevant antigen in 8 of the 10 effusions from patients with well-differentiated ovarian tumors and in 5 of the 11 effusions from patients with poorly differentiated ovarian tumors, whereas the 10 control effusions from patients with various diseases were negative. PMID- 2981615 TI - A serological case-control study of primary hepatocellular carcinoma in Guangxi, China. AB - Guangxi is a very high-risk area for primary hepatocellular carcinoma (PHC); the age-standardized (world population) rates for males and females in that Chinese Autonomous Region were 32.5 and 8.5, respectively. Blood specimens from 50 PHC patients and 50 age- and sex-matched controls in Guangxi were analyzed for hepatitis B surface antigen, antibody to hepatitis B surface antigen, and antibody to hepatitis B core antigen. Eighty-six % of cases were hepatitis B surface antigen positive, compared to 22% of controls (relative risk, 17.0). We estimate from the data that persistent hepatitis B virus infection can account for at least 80% of all PHC cases occurring in Guangxi. The consistency of our findings with those from Hong Kong and Taiwan strongly suggests that hepatitis B virus infection is also an important risk factor for PHC in other parts of southern China. PMID- 2981616 TI - Antiemetic efficacy of levonantradol compared to delta-9-tetrahydrocannabinol for chemotherapy-induced nausea and vomiting. AB - The antiemetic efficacy of im levonantradol, a synthetic cannabinoid, given at a dose of 1 mg every 4 hours, was compared to oral delta-9-tetrahydrocannabinol (THC) given at a dose of 15 mg every 4 hours in a double-blind crossover study. Twenty-six patients receiving emetogenic cancer chemotherapy were evaluated. For each drug, 28% of treated patients had no nausea. The median number of emetic episodes with levonantradol was 2.0 versus 3.0 for THC (P = 0.06). Side effects occurred in 91.7% and 97.3% of levonantradol and THC patients, respectively, with drowsiness and dizziness most commonly seen. Side effects were generally well tolerated, with only 13.9% of levonantradol and 21.6% of THC patients discontinuing treatment because of side effects. Levonantradol appears to be at least as effective an antiemetic as THC and is the only cannabinoid available for parenteral use. PMID- 2981617 TI - Multicenter Phase II trial of etoposide in refractory small cell lung cancer. PMID- 2981618 TI - Pilot study of multiple-fraction daily radiotherapy alternating with chemotherapy in patients with stage IV non-oat cell lung cancer. AB - From March 1982, 31 patients with stage IV non-oat cell lung cancer have been treated. Radiotherapy was given as three 2.00-Gy fractions on Days 1 and 2, 8 and 9, 22 and 23, and 29 and 30, for a total dose of 48 Gy over a 30-32-day treatment period. A three-drug combination of cyclophosphamide (400 mg/m2), doxorubicin (17 mg/m2), and methotrexate (15 mg/m2) was given on Days 3 and 24 and repeated thereafter every 21 days. Three of 31 evaluable patients (10%) achieved objective complete response and 18 of 31 (58%) achieved partial response (ie, regression of 50%-90%), while no change or disease progression was observed in ten of 31 (32%). The overall response rate in our study was 68%, which is a response much higher than other results in extensive disease. However, controlled trials will be necessary to definitively establish the superiority of this regimen to conventional trials. There was a significant shift of performance status towards higher values after treatment: 12 of the 27 patients classified in the 70-80 Karnofsky category before treatment moved to the higher category, 13 remained in the same status, and only two shifted to the worst category, indicating that the treatment had been effective in giving patients a better quality of life during their survival. The median survival was 35 weeks for the entire group of patients and 44 and 15 weeks for the responders and nonresponders, respectively. One of the primary findings of this pilot study was the ability to give one course of 12 Gy of radiation as multiple fractions per day during each of the first 2 weeks of treatment alternated with one course of chemotherapy, with most patients having very mild or no side effects. Giving multiple daily fractions greater than or equal to 4 hours apart permits the delivery of a large amount of irradiation over a short time period (ie, 1-2 days) within the limits of normal tissue toxicity. Increasing the recovery time of radiotherapy by alternation with chemotherapy offers the possibility of increasing the total radiation dose beyond the upper limits now considered feasible by conventional radiation schedules for induction therapy.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981619 TI - Combined intravenous and intra-arterial cyclophosphamide, doxorubicin, and cisplatin (CISCA) in the management of select patients with invasive urothelial tumors. AB - Twenty-eight patients with unresectable urothelial tumors who had no evidence of distant visceral metastasis were managed with iv CISCA (cyclophosphamide, doxorubicin, and cisplatin) and intra-arterial (ia) CISCA chemotherapy. Ten patients had locally advanced disease only and 18 had locally advanced disease with nodal metastasis (ten in pelvic nodes; seven in pelvic and para-aortic nodes; and one in pelvic, para-aortic, and mesenteric nodes). Eleven patients (39%) achieved a complete remission, with a median duration of 49 weeks (range, 25-108). Seven patients (25%) achieved on objective response and ten patients (36%) failed to respond. Iv and ia CISCA chemotherapy is effective in the management of advanced urothelial tumors. Patients with locally advanced disease with or without the presence of nodal metastasis are a select population in whom frequent complete remissions can be achieved. PMID- 2981620 TI - Carboplatin: a very active new cisplatin analog in the treatment of small cell lung cancer. AB - Carboplatin, a cisplatin analog without significant clinical nephrotoxicity, has been evaluated in the treatment of 56 patients with small cell lung carcinoma at a dose of 300-400 mg/m2 iv monthly in a phase II study. Twenty-three patients (41%) achieved a response, including five (9%) complete remissions. Of 30 previously untreated patients, 18 (60%) achieved a response, including three (10%) complete remissions. Median response duration was 4.5 months (range, 2-9). No nephrotoxicity was seen and hydration was not required. Nausea or vomiting occurred in only 24 patients (43%) and was rarely severe. Myelosuppression was dose-limiting: 20 patients (36%) developed leukopenia and eight (14%) developed thrombocytopenia, but leukopenic infections occurred in only three patients. Carboplatin is a very active new agent in the treatment of small cell lung cancer, with less toxicity and better tolerance than cisplatin. It merits further investigation in combination chemotherapy and against non-small cell lung cancer. PMID- 2981621 TI - Phase I trial of 1-(2'-deoxy-2'-fluoro-1-beta-D-arabinofuranosyl)-5-methyluracil (FMAU). AB - 1-(2'-Deoxy-2'-fluoro-1-beta-D-arabinofuranosyl)-5-methyluracil (FMAU), a new pyrimidine nucleoside, is of potential clinical interest both as an anticancer and as an antiviral drug. FMAU is active in vitro and in vivo against P815 and L1210 cell lines resistant to cytarabine. Moreover, in mice inoculated ic with herpes simplex virus Type II, FMAU is 100-fold more potent than vidarabine or acyclovir. We have conducted a phase I trial of FMAU in 17 patients with advanced cancer. The dose levels studied were 2, 4, 8, 16, 32, 64, and 128 mg/m2/day iv for 5 days. The dose-limiting toxic effect was drug-induced central nervous system dysfunction. Although 32 mg/m2/day for 5 days produced only transient, mild symptoms, severe encephalopathy with extrapyramidal dysfunction occurred at 64 and 128 mg/m2/day for 5 days and contributed to two deaths. No toxicity was observed at less than 32 mg/m2. A dose of 32 mg/m2/day for 5 days is suggested for phase II study. Because of its potent and selective antiviral activity, future trials of low doses of FMAU in immunosuppressed patients with herpes virus infections are under consideration. PMID- 2981622 TI - Pharmacokinetics of etoposide in gestochoriocarcinoma. AB - Etoposide (VP16) levels were determined by high-performance liquid chromatography assay in plasma, urine, and surgical specimens of patients with choriocarcinoma undergoing surgery after a dose of 100 or 200 mg/m2 given as a 1-hour infusion. The drug disappeared from plasma biexponentially, with a terminal half-life of 4.1 +/- 0.4 hours, an apparent volume of distribution of 9 +/- 1.1 L/m2, and clearance of 21.5 +/- 3.1 ml/minute/m2. Fifty and 740 minutes after the end of VP16 infusion, the drug concentrations in myometrial carcinoma or in normal myometrium were approximately 40%-50% of those in plasma; 25-180 minutes after the end of drug infusion, the concentrations of VP16 in lung metastases amounted to 19%-43% of those in plasma, whereas in normal lung, the concentrations of VP16 were 38%-61% of those in plasma. In subcutaneous tissue, VP16 levels were much lower than in plasma (about 5%-27%). About 50% of VP16 was eliminated in the urine as unchanged drug, glucuronide, or sulfate. PMID- 2981624 TI - Transcriptional block caused by a negative supercoiling induced structural change in an alternating CG sequence. AB - Using supercoiled plasmids containing a (CG)16 sequence downstream of a promoter, it is shown that purified E. coli RNA polymerase can transcribe through the sequence when it is in the B helical form. However, the polymerase together with its nascent transcript is blocked at the boundary of the CG sequence proximal to the promoter when the template is negatively supercoiled to flip the CG sequence to the left-handed Z-form. S1 nuclease mapping of in vivo transcripts from an E. coli gyrase temperature-sensitive mutant harboring the plasmids indicates that the bulk of the transcripts at either permissive or nonpermissive temperatures can proceed through the CG sequence, suggesting that the sequence is normally in the B helical form in vivo. The almost total blockage of transcription in vitro by the (CG)16 sequence in a highly negatively supercoiled DNA is not observed for a d(CA)21 X d(TG)21 insert. PMID- 2981623 TI - Intracellular pathways of receptor-bound GnRH agonist in pituitary gonadotropes. AB - Localization of GnRH receptors in rat pituitary gonadotropes was studied by use of 125I-[azidobenzoyl-D-Lys6]GnRH which, upon photolysis, is covalently bound to the receptor molecule. Using high resolution autoradiography, it was found that, after a 90-min incubation of the analog with pituitary cells at 4 degrees C, 93% of the silver grains were associated with the plasma membrane of the gonadotropes. After 45-min incubation of the cells at 37 degrees C, clustering and internalization of the receptor-bound GnRH analog were evident. Silver grains were associated with coated pits, intracellular vesicles, Golgi complexes, lysosome-like structures and secretory granules. The data indicate that receptor bound GnRH agonist is internalized, at least in part, via coated pits and is subsequently routed to lysosomes where degradation of the hormone-receptor complex may occur. The presence of a considerable amount of silver grains associated with secretory granules may suggest that some of the internalized receptor molecules can escape degradation and be recycled to the cell membrane. PMID- 2981625 TI - Recombination site selection by Tn3 resolvase: topological tests of a tracking mechanism. AB - In vitro recombination by Tn3 resolvase of plasmids containing two directly repeated recombination (res) sites generates two singly interlinked catenated rings. This simple product catenane structure was maintained over a wide range of substrate supercoil densities and in a reaction mixture in which phage lambda Int mediated recombination generated its characteristic multiply interlinked forms. Using substrates containing four res sites, we found that resolvase recombined neighboring res sites with high preference. This position effect implies that resolvase searches systematically along the DNA for a partner site. Intervening res sites in the opposite orientation did not prevent translocation. We analyzed the geometric arrangement of the interlocked rings after multiple recombination events in a four-site substrate and the pattern of segregation of nonspecific reporter rings catenated to the standard substrate. The results of these novel topological tests imply that the translocating enzyme may not make continuous contact with the DNA. PMID- 2981626 TI - Autoregulation of the DNA replication gene dnaA in E. coli K-12. AB - The dnaA gene in E. coli K-12 is required for the initiation of DNA replication. Although the specific function of the dnaA protein is unknown, it has been suggested that it is a regulator of the frequency of initiation. In this paper we report that the expression of both a dnaA-lacZ translational fusion and a dnaA trpA-lacZ transcriptional fusion in vivo are sensitive to changes in the level of functional dnaA protein. Overproduction of the dnaA gene product leads to a reduction in expression from both fusions while introduction of dnaA- alleles results in an increased expression. Results from a deletion analysis of the dnaA promoter/regulatory region suggest that both dnaA promoters are regulated by the dnaA gene product and that a site between the two promoters is responsible for the regulation. DNAase protection experiments showed that the dnaA protein binds to DNA in the region of the two dnaA promoters. Our results indicate that the dnaA gene product regulates its own synthesis by inhibiting transcription from both of its promoters. PMID- 2981627 TI - Differential expression of photosynthesis genes in R. capsulata results from segmental differences in stability within the polycistronic rxcA transcript. AB - We report that the light-harvesting and reaction center genes in the rxcA locus of R. capsulata are contained within a single operon and that their differential expression results predominantly from marked segmental differences in stability within the polycistronic rxcA transcript. The 3' portion of this transcript is rapidly degraded to give rise to either of two slowly decaying mRNA remnants, both of which encode only the light-harvesting polypeptides. The greater stability of these remnants accounts for nearly all of the difference between the concentrations of the light-harvesting and reaction center proteins. The unstable 3' portion of the transcript is delimited by two alternative stem-and-loop structures, which apparently act as barriers to 3' exoribonucleases and thereby protect the upstream RNA segment. When a DNA fragment containing the rxcA locus was fused to a plasmid promoter and transcribed in E. coli, the long precursor transcript was processed to two short messages of greater stability, as in R. capsulata. PMID- 2981628 TI - Functional homology of mammalian and yeast RAS genes. AB - Yeast spores lacking endogenous RAS genes will not germinate. If such spores contain chimeric mammalian/yeast RAS genes or even the mammalian H-ras gene under the control of the galactose-inducible GAL10 promoter, they will germinate in the presence of galactose and produce viable haploid progeny dependent on galactose for continued growth and viability. These results indicate that the biochemical function of RAS proteins is essential for vegetative haploid yeast and that this function has been conserved in evolution since the progenitors of yeast and mammals diverged. PMID- 2981629 TI - The appearance and internalization of transferrin receptors at the margins of spreading human tumor cells. AB - Using gold complexes stabilized with a monoclonal antibody specific for the human transferrin receptor, the distribution of transferrin receptors on the surfaces of human epidermoid carcinoma A431 cells has been mapped at high resolution. On prefixed cells and cells incubated at 5 degrees C, the receptors are predominantly within and around clathrin-coated microdomains near the free cell margin. By preincubating the cells with saturating concentrations of free antibody at 5 degrees C and warming them to 37 degrees C in the presence of the gold complexes, the appearance of new receptors in the membrane has been followed. The majority first appear near the free cell margin and then move centripetally. At first, they are monodisperse, but as they move toward the site of internalization they form loose aggregates. Within the immediate vicinity of the clathrin-coated microdomains the migrating receptors form closely packed, ordered aggregates. These observations indicate recycling transferrin receptors move to their site of internalization without cross-linking. PMID- 2981631 TI - opa: a novel family of transcribed repeats shared by the Notch locus and other developmentally regulated loci in D. melanogaster. AB - The principal transcription product of Notch, a locus involved in the neurogenesis of D. melanogaster, is a developmentally regulated poly(A)+ RNA approximately 10.5 kb in length. Analysis of the structure of this RNA has revealed a 93 bp repeated sequence that is shared by many other developmentally regulated transcription units. Nucleotide sequence analysis of the repeat shows an unusual structure consisting predominantly of the triplets CAG and CAA, both of which can code for the amino acid Gln. We present evidence indicating that the Notch repeat is a member of a novel family of repetitive elements, which we term the opa family. Our data suggest that some of these elements may be not only transcribed but also translated. We compare opa with other known transcribed repeats and speculate on its functional significance. PMID- 2981630 TI - In yeast, RAS proteins are controlling elements of adenylate cyclase. AB - S. cerevisiae strains containing RAS2val19, a RAS2 gene with a missense mutation analogous to one that activates the transforming potential of mammalian ras genes, have growth and biochemical properties strikingly similar to yeast strains carrying IAC or bcy1. Yeast strains carrying the IAC mutation have elevated levels of adenylate cyclase activity. bcy1 is a mutation that suppresses the lethality in adenylate cyclase deficient yeast. Yeast strains deficient in RAS function exhibit properties similar to adenylate cyclase deficient yeast. bcy1 suppresses lethality in ras1- ras2- yeast. Compared to wild-type yeast strains, intracellular cyclic AMP levels are significantly elevated in RAS2val19 strains, significantly depressed in ras2- strains, and virtually undetectable in ras1- ras2- bcy1 strains. Membranes from ras1- ras2- bcy1 yeast lack the GTP-stimulated adenylate cyclase activity present in membranes from wild-type cells, and membranes from RAS2val19 yeast strains have elevated levels of an apparently GTP independent adenylate cyclase activity. Mixing membranes from ras1- ras2- yeast with membranes from adenylate cyclase deficient yeast reconstitutes a GTP dependent adenylate cyclase. PMID- 2981632 TI - Molecular probes define different regions of the mouse t complex. AB - Four genomic clones obtained from microdissected fragments of the proximal portion of mouse chromosome 17 have been used to identify a series of t-haplotype specific restriction fragments. Their specificity is defined by presence in eight complete t haplotypes and absence from 18 inbred strains of wild-type mice. Partial t haplotypes contain subsets of the t-specific fragments, and each can be classified according to the t-specific fragments it contains. This is the first molecular evidence that independent partial t haplotypes contain different lengths of t haplotype DNA. Recombination studies indicate that partial t haplotypes suppress recombination in proportion to the extent of t haplotype DNA they contain. Molecular analysis of partial t haplotypes shows that the t specific fragments map to and thus define different regions of the t complex. Certain regions of t haplotype DNA defined by t-specific restriction fragments can be correlated with loci involved in the control of transmission ratio distortion. PMID- 2981633 TI - Transposition of the immunoglobulin heavy chain enhancer to the myc oncogene in a murine plasmacytoma. AB - A novel mechanism of oncogene activation by transposition of a tissue-specific cellular enhancer is described. A rearranged c-myc oncogene was cloned from murine plasmacytoma ABPC17 in the expectation that it would reflect the t(6;15) chromosome translocation carried by this tumor. The rearrangement instead reflects an insertion 361 bp 5' to the c-myc gene on chromosome 15. The insert conveys a 2.3 kb segment of the immunoglobulin heavy (H) chain locus from chromosome 12. Since the insertion introduces the lymphoid-specific enhancer from the JH-Smu region and also disrupts a region implicated in normal c-myc control, it may account for the c-myc transcription observed in ABPC17. The structure of the transposed segment and a corresponding deletion in the JH-Smu region suggests that the transposition reflects a complex recombination between chromosomes 15 and 12. Since the t(6;15) breakpoint is not near c-myc, chromosome 15 must have undergone an independent exchange with chromosome 6. PMID- 2981634 TI - Intron/exon structure of the chicken pyruvate kinase gene. AB - The chicken pyruvate kinase gene is interrupted by at least ten introns, including nine introns within the coding region. We compare the structure of this gene with the three-dimensional protein structure of the homologous cat muscle enzyme. The introns are not randomly placed--they divide the coding sequence into fairly uniformly sized pieces encoding discrete elements of secondary structure. The introns tend to fall at interruptions between stretches of alpha-helix or beta-sheet residues, and each of the six exons that contribute to the barrel shaped central domain include one or two repeats of a simple unit, an alpha-helix plus a beta strand. This structure suggests that introns were not inserted into a previously uninterrupted coding sequence, but instead are products of the evolution of the first pyruvate kinase gene. We have found some sequence homology between a segment of pyruvate kinase and the structurally homologous mononucleotide binding fold of alcohol dehydrogenase. The superposition of these two regions aligns an intron from the maize alcohol dehydrogenase gene four nucleotides from an intron in the chicken pyruvate kinase gene. PMID- 2981635 TI - Nucleotide sequence of the AIDS virus, LAV. AB - The complete 9193-nucleotide sequence of the probable causative agent of AIDS, lymphadenopathy-associated virus (LAV), has been determined. The deduced genetic structure is unique: it shows, in addition to the retroviral gag, pol, and env genes, two novel open reading frames we call Q and F. Remarkably, Q is located between pol and env and F is half-encoded by the U3 element of the LTR. These data place LAV apart from the previously characterized family of human T cell leukemia/lymphoma viruses. PMID- 2981637 TI - Remote regulatory sequences of the Drosophila glue gene sgs3 as revealed by P element transformation. AB - We have examined the transcriptional activity of the Formosa sgs3 glue gene reinserted in the Drosophila genome in transposons containing various arrangements of its natural flanking sequences. The shortest transposons, retaining 127 bp of 5' sequence show no (or very rare) expression. Constructs with up to 1.4 kb of 5' sequence show tissue- and stage-specific accumulation of transcripts, but at severely reduced levels when compared with the resident sgs3 glue gene. To obtain wild-type levels of transcripts, sequences contained in the next 1.3 kb are necessary. At least one regulatory element defined by these experiments is bi-directional. Our results show that marked P-element transformation vectors can be used to analyze regulatory elements of linked genes. PMID- 2981636 TI - A fraction of the mouse genome that is derived from islands of nonmethylated, CpG rich DNA. AB - About 1% of the mouse genome is cleaved by Hpa II to give a discrete fraction on gels. The nonmethylated fraction is present in all tested tissues, including sperm, and contains Hpa II sites at about 15 times their frequency in bulk DNA. About 80% of the fraction is composed of sequences that occur once or a few times per genome; the remainder is largely rDNA. Unlike bulk DNA, the fraction is not deficient in CpG, and this may be directly due to the lack of methylation. Genomic mapping of three nonribosomal fragments showed that they are part of islands of DNA within which nonmethylated Hpa II and Hha I sites are highly concentrated. We estimate about 30,000 islands per haploid genome and discuss evidence that many may be associated with genes. PMID- 2981638 TI - Selective inhibitory effect of Hu-IFN-gamma on the agarose clonability of tumor derived lymphoid cell lines. AB - Recombinant human interferon alpha (IFN-alpha) and interferon gamma (IFN-gamma) were compared for their ability to influence the proliferative capacity of tumor derived cell lines and of normal B lymphocytes infected in vitro by Epstein-Barr virus (EBV). EBV-induced B-cell proliferation was suppressed almost completely when 10(2) U/ml IFN-alpha were added to the culture medium while the same dose of IFN-gamma had significantly lower inhibitory activity. The pure IFNs differed in their ability to influence the growth of three Burkitt lymphoma-derived cell lines, Raji, Daudi, and Namalwa, depending on whether the cells were propagated in suspension or in semisolid cultures. IFN-alpha inhibited cell proliferation under both culture conditions with thresholds of sensitivity characteristics for each cell line. In contrast, IFN-gamma had no effect on the growth in suspension but it abolished the clonogenic potential of tumor cell lines in semisolid agarose. The results suggest that the two IFN types may exert their growth inhibitory activity through different mechanisms of action. PMID- 2981639 TI - Inhibition of proliferation without affecting the generation of cytotoxicity in the human mixed lymphocyte reaction. AB - Phosphoramide mustard (PM) is considered to be the major tumoricidal metabolite of cyclophosphamide in vivo. The effects of this metabolite in vitro on several immune functions of human lymphocytes have been investigated. Very low concentrations (10(-7) to 10(-9) M) of PM added to lymphocyte cultures inhibited proliferation of the lymphocytes in response to mitogens and alloantigens. At these concentrations, inhibition of proliferation appeared to be due to a direct action of PM on the proliferative cells. Thus, concanavalin A-stimulated lymphocytes still acquired IL-2 receptors (Tac antigen) normally in the presence of PM (10(-6) to 10(-9) M). Only exceedingly high concentrations of PM (10(-5) M or greater) prevented the acquisition of Tac antigen. Similarly, the inhibition of proliferation was probably not related to endogenous IL-2 levels: addition of exogenous IL-2 to PM-containing cultures did not result in any restoration of proliferation. Further evidence that PM directly affected proliferative cells was that low concentrations of PM inhibited the proliferation of T cells continuously growing in IL-2. The exposure time to PM necessary for inhibition was essentially identical to those for lymphoproliferative responses to mitogens and alloantigens. Paradoxically, however, the generation of cytotoxic lymphocytes in mixed lymphocyte reactions (MLRs) and mixed lymphocyte tumor cell cultures (MLTCs) was very resistant to PM. In parallel MLRs and MLTCs the cytotoxic responses were resistant to approximately 1000-fold more PM than were the proliferative responses. Only at 10(-5) M PM were these inhibited. These data suggest that clonal expansion of cytotoxic lymphocytes or their precursors by proliferation is not an absolute requirement for the generation of cytolytic activity. PMID- 2981640 TI - Kinetics of activation antigen expression by in vitro-stimulated human T lymphocytes. AB - In this study a panel of monoclonal antibodies was used to investigate the kinetics of the appearance of activation-linked surface determinants as well as cytoplasmic and nuclear determinants in human T cells following lectin stimulation. Well known activation markers, such as Ia/DR, transferrin receptor, IL-2 receptor, T10, and gp24, were compared and investigated together with the T13 structure, recently found in this laboratory. T13, not demonstrable on resting T cells, could be seen within 24 hr after lectin stimulation. Kinetics of the appearance were similar to IL-2 receptor and transferrin receptor expression. Ia/DR synthesis was investigated separately for each polypeptide and the cytoplasmic invariant gamma-chain expression could be demonstrated for the first time with a gamma-chain-specific monoclonal antibody VIC-Y1. Moreover, gamma chain synthesis seems to precede alpha- and beta-chain occurrence in human T cells. In addition, data from quantitative studies on antigenic densities are presented. PMID- 2981641 TI - Suppression by cyclosporin A of murine T-cell-mediated immunity against viruses in vivo and in vitro. AB - The immunosuppressive effect of Cyclosporin A on T-cell-mediated antiviral immune responses was examined. When administered intraperitoneally CS-A abrogated anti vaccinia virus, anti-lymphocytic choriomeningitis virus (LCMV), and anti vesicular stomatitis virus (VSV) T-cell responses in a dose-dependent fashion. Usually 50-60 mg/kg were efficient in suppressing primary T-cell responses completely. In contrast, 10-20 mg/kg often enhanced T-cell responses significantly when compared with controls. Suppression was observed if CS-A treatment was started before virus injection and up to 12 hr after infection; CS A given 24 hr after the virus still suppressed T-cell activity partially. A 50 mg/kg dose of CS-A suppressed secondary anti-vaccinia virus or anti-VSV T-cell responses in vivo by a factor of about 10. This dose suppressed the primary T cell-dependent footpad swelling induced by local LCMV infection and prevented T cell-mediated immunopathological death due to LCM when LCMV was injected intracerebrally. In addition, clearance of LCMV was delayed drastically by CS-A treatment. When added to cultures of in vivo-primed antiviral T cells that were restimulated in vitro, CS-A inhibited both proliferation as well as generation of virus-specific cytotoxic T cells in a dose-dependent way. The results show that in CS-A-treated mice primary and secondary antiviral T-cell responses are strongly inhibited; acute viral infections with cytopathic viruses may therefore be more dramatic. In contrast immunopathological T-cell-mediated disease caused by noncytopathic viruses such as LCMV may be prevented or attenuated. PMID- 2981642 TI - Natural killer activity against cultured human neural tumor and fetal brain cells. AB - Ten human neural tumor lines and three established from normal human brain were analyzed for sensitivities to natural killer (NK) cytolysis. Compared to MOLT-4, fetal brain cells were sensitive, but those from adult brain and eight of ten neural tumor cell lines demonstrated marked NK resistance. The frequencies of target-binding cells (TBC) and single-cell lysis of glioma cells bound within tumor cell conjugates demonstrated that the resistance of two lines was explained either by a decrease in the frequencies of TBC or reduced ability of bound NK cells to lyse the tumor cell conjugates. A third resistant line demonstrated decreases in both TBC and tumor cell conjugate lysis. Two glioma lines with less NK resistance had greater frequencies of TBC or conjugate lysis than the resistant lines. Thus, NK resistance can result from decreased recognition of targets, diminished NK lysis of bound targets, or a combination of both. PMID- 2981643 TI - Fluoro-substituted N-nitrosamines. 7. Non-genotoxic N-nitroso-bis(2,2,2 trifluoroethyl)amine and N-nitroso-bis(2,2,3,3,4,4,4-heptafluorobutyl)amine: binding to cytochrome P-450, acidity of alpha-protons and pharmacokinetic investigations. AB - The biologically inactive fluorinated nitrosamines N-nitroso-bis(2,2,2 trifluoroethyl)amine (NDEA-F6) and N-nitroso-bis-(2,2,3,3,4,4,4 heptafluorobutyl)amine (NDBA-F14) were investigated for binding affinity to cytochrome P-450 and for ease of alkali-induced proton abstraction at the alpha-C atom, in comparison with biologically active analogues (N-nitroso-diethylamine, NDEA; N-nitroso-2,2,2-trifluoroethylethylamine, NDEA-F3; N-nitrosodibutylamine, NDBA; N-nitroso-4,4,4-trifluorobutylbutylamine, NDBA-F3; N-nitroso-bis(4,4,4 trifluorobutyl)amine, NDBA-F6). Binding to cytochrome P-450 was studied by spectroscopic measurements (optical difference spectra with microsomal fractions); base-catalyzed deuterium exchange of alpha-hydrogen atoms was followed by 1H n.m.r. measurements. Additionally the excretion of NDEA-F6 and NDBA-F14 in expired air, urine and faeces was studied after oral application to the rat. Compared with the biologically active nitrosamine analogues, NDEA-F6 and NDBA-F14 showed higher binding affinity to cytochrome P-450. N.m.r. spectroscopy showed that NDEA-F6, NDBA-F14 and NDEA-F3 (at the fluorinated alkyl chain) were rapidly deprotonated at the alpha-C-position in sodium perdeutero methylate, in contrast to the other analogues tested. In vivo, NDEA-F6 and NDBA-F14 were excreted unchanged, mainly via exhalation. The biological inactivity of NDEA-F6 and NDBA-F14, together with the observed blocking of their microsomal activation can be reconciled with the experimental findings which indicate that homolytic alpha-C-H bond fission is more likely to be involved in alpha-C-hydroxylation of dialkylnitrosamines, than alpha-proton abstraction. PMID- 2981644 TI - Two distinct effects of oxygen on vascular tone in isolated porcine coronary arteries. AB - The relation between PO2 and vessel tone was studied in isolated porcine left descending coronary artery rings. Porcine left descending coronary artery mounted isometrically and equilibrated in Krebs-bicarbonate solution (37 degrees C, pH 7.4, when gassed with 95% oxygen + 5% carbon dioxide) exhibited spontaneous basal tone. Decreasing bath PO2 to 40%, 20%, and 12% elicited sustained increases in basal tension which were reversible, ranging between 10% and 20% of the contraction induced by 40 mM potassium chloride. Further decreases in PO2 to near zero (anoxia) resulted in relaxation to baseline. Cyclooxygenase inhibitors indomethacin (5.5 X 10(-6) M), aspirin (5 X 10(-5) M), and meclofenamate (10(-5) M) decreased vascular tone and totally prevented coronary vasoconstriction induced by lowering bath PO2 to 12% or 40% but did not affect anoxic vasorelaxation. Neither basal tone nor the vasoconstriction induced by decreases in bath PO2 were influenced by the antihistaminergic drug pyribenzamine (10(5) M) or by the alpha-adrenergic blocker phentolamine (10(-6]. Isoproterenol (10(-9) to 10(-8) M) or an elevation of the bath potassium concentration from 5.9 to 11 mM significantly augmented coronary vasoconstriction induced by lowering bath PO2 from 95% to 40%. Elevation of the bath potassium chloride concentration to 40 mM further increased isometric force but inhibited the vasoconstriction in response to decreasing PO2 from 95% to 40%. Anoxia relaxed contractions induced by 40 mM potassium chloride, histamine, or ouabain. The data suggest the existence of two distinct oxygen-sensitive mechanisms in porcine coronary arteries, both of which regulate vascular tone. One is activated at relative high PO2 values (10-40%), and the vasoconstriction induced by this mechanism is mediated by vascular prostaglandin synthesis. The other is expressed at low PO2 values (near zero), and the depression of mechanical activity by this mechanism may be related to limitation of oxidative energy metabolism. The first mechanism can be augmented by beta-adrenoceptor stimulation indicative of an interaction between vascular prostaglandin synthesis and beta-adrenergic mechanisms in the coronary artery wall. PMID- 2981645 TI - Vascular adrenergic neuroeffector function does not decline in aged rats. AB - To investigate adrenergic control of blood vessels during aging, rats aged 6, 12, 20, and 27 months were studied using in vitro techniques. Accumulation of [3H]norepinephrine, one index of adrenergic nerve density, did not alter with age in the femoral or renal arteries or renal vein. In the femoral vein [3H]norepinephrine accumulation was greater at 6 and 27 months of age. Norepinephrine sensitivity was determined in both an innervated vessel, the femoral artery, and a non-innervated vessel, the carotid artery. In both cases, sensitivity to norepinephrine did not alter with age. In the renal and femoral arteries and veins, no significant changes in maximum responses to norepinephrine (10(-5) M), potassium chloride, or transmural nerve stimulation were seen with advancing age. Furthermore, frequency response curves (2-16 Hz, 200 pulses) did not differ with age for any of the four vessels studied, with one exception. The response to stimulation at 4 Hz of the femoral vein from 6-month-old rats was significantly larger than responses at other ages. During nerve stimulation, the renal vein exhibited rapid contractions superimposed upon the maintained contractile response. This type of rapid contraction occurred only rarely (1 out of 5) in the renal vein from 27-month-old rats. In summary, neither adrenergic nerve density as reflected by [3H]norepinephrine accumulation nor norepinephrine sensitivity decline with age. As the net effect of various components, the ability of vascular smooth muscle to respond to adrenergic nerve stimulation is also maintained during advancing age. PMID- 2981646 TI - Inhibitors of prostaglandin synthesis augment beta-adrenergic responsiveness in canine coronary arteries. AB - The effects of inhibition of endogenous prostaglandin synthesis on the release of norepinephrine from sympathetic nerves and on postjunctional adrenergic responsiveness were studied in isolated canine left circumflex coronary arteries. In rings, suspended for isometric tension recording and contracted with prostaglandin F2 alpha, transmural electrical stimulation caused frequency dependent relaxations, which were blocked by propranolol and augmented by indomethacin. In superfused strips, previously incubated with [3H]norepinephrine, electrical stimulation (2 Hz) increased the overflow of tritiated neurotransmitter; indomethacin did not influence basal or evoked [3H]norepinephrine overflow. Exogenous norepinephrine caused relaxations in rings contracted with prostaglandin F2 alpha, but increases in tension in potassium depolarized tissues which could be abolished by phentolamine; isoproterenol induced relaxations in both cases. Indomethacin significantly augmented the relaxation in response to exogenous norepinephrine (during contractions with prostaglandin F2 alpha) and reversed norepinephrine-induced contractions (during potassium-depolarization) into relaxation. Other cyclooxygenase inhibitors had comparable effects. In the presence of propranolol, indomethacin did not diminish contractions evoked by norepinephrine in depolarized rings. Relaxations induced by sodium nitroprusside or acetylcholine during contractions caused by prostaglandin F2 alpha or potassium chloride were not affected by indomethacin. The augmentation of beta-adrenergic responsiveness by indomethacin was abolished by exogenous prostacyclin. The prostacyclin synthetase inhibitor tranylcypromine and exogenous prostaglandin E2 depressed beta-adrenergic responsiveness. Indomethacin did not affect the facilitatory action of phosphodiesterase inhibition on beta-adrenergic relaxation. The data suggest that endogenous prostaglandins (most probably prostacyclin and prostaglandin E2) exert a "braking" effect on beta-adrenergic responsiveness in coronary arterial smooth muscle. PMID- 2981647 TI - Relative inhibition of human adenylate kinase and creatine kinase isoenzymes by adenosine 5'-monophosphate and diadenosine pentaphosphate. PMID- 2981648 TI - Precipitation of Mn2+ with HCO3- in determinations of high-density lipoprotein subfractions. PMID- 2981649 TI - Stimulus-dependent inhibition of superoxide generation by prostaglandins. AB - Infiltrating phagocytes generate superoxide anion (O2-) and prostaglandin (PG) at sites of inflammation. Thus PG-O2- interactions may be important to the initiation and control of inflammation. PGE1, PGE2, and PGD2 inhibit O2- generation (as measured by superoxide dismutase-inhibitable reduction of ferricytochrome c) in a dose-dependent manner (10(-6)-10(-9) M) when human peripheral blood polymorphonuclear leucocytes (PMN) are stimulated with 10(-7) M of the chemotactic peptide, N-formyl-methionyl-leucyl-phenylalanine (FMLP). These PG did not alter O2- generation when PMN were stimulated with 0.1 microgram/ml phorbol myristate acetate (PMA) or 1 mg/ml serum-treated zymosan (STZ). Increments of cyclic AMP (cAMP) (peak: fourfold) in PGE1, PGE2, and PGD2 treated PMN stimulated with PMA or STZ (in which O2- was not reduced) were similar to those in PG-treated PMN stimulated with FMLP (in which O2- was reduced markedly). High concentrations of theophylline and dibutyryl cAMP reduced FMLP and STZ stimulated O2- generation but had no effect on PMA stimulation, suggesting that the stimuli induce different sensitivities to the effects of cellular cAMP. PGF2 alpha had little effect on O2- generation or cAMP levels regardless of the stimulus. PGE1 did not inhibit binding of FML(3H)P to PMN and did not scavenge O2 anions. Therefore the effect of PG on O2- production is dependent on the specific stimulator and an increased concentration of cAMP in activated PMN is by itself not sufficient to limit O2- generation induced by all stimuli. PMID- 2981651 TI - Optimal conditions for measurement of Na+,K+-ATPase activity of human leucocytes. AB - The Na+,K+-ATPase activity of human leucocytes was assayed by measuring the release of inorganic phosphate (Pi) from ATP. The maximum enzyme activity was achieved under the following conditions: concentration (mmol/l), Tris/HCl 50, Na 100, K 15, ATP 5, Mg 7, EDTA 1; pH 7.2 and temperature 37 degrees C, were optimal. Ouabain showed maximal inhibition at a concentration of 10-100 mumol/l. Ethanol, the solvent for ouabain, had a dose-related inhibitory effect. Heparin or citrate used as an anticoagulant gave similar results. Leucocyte samples could be stored at -20 degrees C for up to 6 days without loss of activity. Hypotonic lysis had advantages over sonication as the technique for cell disruption. The leucocyte Na+,K+-ATPase enzyme activity in healthy subjects was 186 mumol of Pi h 1g-1 of protein (median) with a range 136-243 mumol of Pi h-1g-1 of protein. The within-batch coefficient of variation was 6.4% and the between-batch precision was 9.6%. PMID- 2981650 TI - Mast cell-dependent inflammatory mediators and their putative role in bronchial asthma. PMID- 2981653 TI - Aseptic meningitis in a neonate with an oral vesicular lesion. AB - Enteroviruses may be responsible for severe, life-threatening diseases, such as meningitis, within the neonatal period. Differentiation of an enteroviral etiology from that of herpes simplex virus may be difficult initially. We describe a case of aseptic meningitis in which the use of a viral blood culture was helpful. PMID- 2981652 TI - Cardiovascular properties of metkephamid, a delta opioid receptor agonist, in man. AB - Opioid receptors exist in at least three forms: mu, delta and kappa. Agonists at mu receptors produce orthostatic hypotension in man by a mechanism involving a reduction in baroreflex sensitivity. We describe here the cardiovascular properties of metkephamid, a relatively selective delta opioid receptor agonist. Blood pressure, heart rate and plasma noradrenaline concentration were measured over a 7 h period in eight normal young male volunteers in the supine position and after 70 degrees 5 min head-up tilt, after receiving metkephamid (50 mg intramuscularly) or placebo. Metkephamid increased heart rate in the supine position with no change in blood pressure or plasma noradrenaline concentration. This was accompanied by symptoms consistent with an anti-muscarinic anticholinergic effect. Head-up tilt resulted in substantial hypotension after metkephamid with an attenuated change in heart rate and no increase in noradrenaline concentration. We conclude that delta as well as mu opioid receptor agonists can produce orthostatic hypotension with attenuation of heart rate response. Metkephamid possesses anticholinergic properties not seen with mu receptor agonists, suggesting a possible role of delta opioid receptors in cholinergic activity. PMID- 2981654 TI - A comparison of the polypeptide composition of plasma membranes prepared from the white adipose tissue and adipocytes of the mouse, rat, rabbit, ox and chicken by a Percoll self-forming gradient procedure. AB - The Percoll self-forming gradient method has been applied for the preparation of plasma membranes from white adipose tissue and isolated adipocytes from mouse, rat, rabbit, ox and chicken. Characterization of the membranes produced by marker enzyme analysis confirmed the general applicability of the method. The polypeptide composition of the membranes was studied by polyacrylamide gel electrophoresis. The patterns seen with tissue and adipocytes of a single species source were compared and the Coomassie blue and periodic acid Schiff staining bands characteristic of each species were compared systematically. Additionally materials from lean (FA/-) and obese (fa/fa) Zucker rats were compared. The species-specific differences in polypeptide composition form a useful basis for the further development of methods whereby the adipocyte-differentiated phenotype may be detected in a wide range of systems. PMID- 2981655 TI - Characterization of gill (Na+ + K+)-ATPase in the sea bass (Dicentrarchus labrax L.). AB - Bass gill microsomal preparations contain both a Na+, K+ and Mg2+-dependent ATPase, which is completely inhibited by 10(-3)M ouabain and 10(-2)M Ca2+, and also a ouabain insensitive ATP-ase activity in the presence of both Mg2+ and Na+. Under the optimal conditions of pH 6.5, 100 mM Na+, 20 mM K+, 5 mM ATP and 5 mM Mg2+, (Na+ + K+)-ATPase activity at 30 degrees C is 15.6 mumole Pi hr/mg protein. Bass gill (Na+ + K+)-ATPase is similar to other (Na+ + K+)-ATPases with respect to the sensitivity to ionic strength, Ca2+ and ouabain and to both Na+/K+ and Mg2+/ATP optimal ratios, while pH optimum is lower than poikilotherm data. The enzyme requires Na+, whereas K+ can be replaced efficiently by NH+4 and poorly by Li+. Both Km and Vm values decrease in the series NH+4 greater than K+ greater than Li+. The break of Arrhenius plot at 17.7 degrees C is close to the adaptation temperature. Activation energies are scarcely different from each other and both lower than those generally reported. The Km for Na+ poorly decreases as the assay temperature lowers. The comparison with literature data aims at distinguishing between distinctive and common features of bass gill (Na+ + K+)-ATPase. PMID- 2981657 TI - Compared evolution of plasma fibronectin and angiotensin-converting enzyme levels in septic ARDS. AB - Serum angiotensin converting enzyme (serum ACE) levels and plasma fibronectin levels were measured daily in 46 septic patients during a ten day period. Thirty eight patients developed ARDS; 28 survived (group 1), ten died (group 2), eight patients had no features of ARDS and survived (group 3). Sequential measurements of ACE and fibronectin levels were compared and plotted against indexes of respiratory impairment: PaO2 max Qs/Qt, static compliance and VD/VA ratio. These indexes were taken as criteria of weaning from controlled ventilation. During ARDS (groups 1 and 2), serum ACE levels decreased and were closely correlated with the severity of lung injury. Persistently decreased levels after eight days were consistent with continuing injury or lack of endothelial repair. On the other hand, plasma fibronectin levels increased throughout the study in survivors (group 1 and 3) and decreased in the group with fatal ARDS only (group 2). These results indicate that serum ACE levels might be a good index of endothelial injury and repair during ARDS and fibronectin a better index for evolution of sepsis and vital prognosis. PMID- 2981656 TI - Effect of phentolamine in controlling temperature and acidosis associated with cardiopulmonary bypass. AB - This study assessed the effects of phentolamine on rewarming patterns and metabolic acidosis in 37 patients subjected to hypothermia during cardiopulmonary bypass for the performance of aortocoronary bypass grafting. An additional 16 patients undergoing the same surgery received no phentolamine and served as a control group. In all patients, sodium bicarbonate (44.6 mEq) was administered only when the negative base excess was 3.0 mEq/L or greater. Sixty-eight percent of the patients receiving phentolamine and 56% of the control patients exhibited a uniform rewarming pattern in which the rectal, hand, and foot temperatures increased in parallel. In 32% of the patients receiving phentolamine and in 44% of the control patients, rectal and hand temperatures increased more than foot temperature during rewarming. Analysis of base excess values in the subgroups of patients with similar rewarming patterns indicated that base deficits were significantly decreased in patients receiving phentolamine. Phentolamine administration was also associated with significantly lower blood lactate levels and sodium bicarbonate requirements, as well as improvements in overall appearance and mental status. These data suggest that the routine use of phentolamine in patients undergoing cardiopulmonary bypass may be associated with more uniform body cooling and rewarming and improved tissue perfusion. PMID- 2981658 TI - Pulmonary complications of bone marrow transplantation. AB - Bone marrow transplantation (BMT) for hematologic disorders is potentially curative in selected persons. These patients may be immunocompromised for months after engraftment as a consequence of chemotherapy, irradiation, acute and chronic graft-vs-host disease (GVHD), and maturing recipient marrow. Pulmonary complications commonly occur during the early and late periods after BMT and are associated with significant morbidity and mortality. The leading early-onset complication is interstitial pneumonitis, most commonly associated with cytomegalovirus infection but also related to possible toxicities from chemotherapy and irradiation. Major late-onset problems include bacterial sinopulmonary infections and obstructive airway disease thought to be associated with chronic GVHD. The exact mechanisms of lung injury are probably quite complex, and unfortunately, often cause irreversible pulmonary disease, even in the patient who has had successful transplantation. Antimicrobial prophylaxis, modified chemotherapy and irradiation dosages, and antiviral immunization have been shown to reduce the incidence of early-onset pulmonary problems. Early recognition and treatment of late-onset problems will, it is hoped, minimize respiratory limitations. PMID- 2981660 TI - CDC Diabetes Control Programs--overview of diabetes patient education. PMID- 2981659 TI - Automatic cell identification and enrichment in lung cancer: V. Adenocarcinoma and large cell undifferentiated carcinoma. AB - The aims of this study were to develop a protocol for the identification and enrichment of cancer cells from sputum obtained from patients with adenocarcinoma of the lung (n = 6) and large-cell undifferentiated carcinoma of the lung (n = 2), and to compare these findings with the results from our previous studies on other cell types from lung cancer. The hypotheses tested were: Cancer cells in sputum can be preserved following flow sorting. Enrichment for cancer cells from acridine orange (AO)-stained specimens can be achieved. Discrimination of cancer cells from noncancer cells is by AO green fluorescence and discrimination of lymphocytes from other cell types is by AO red fluorescence. Cancer cells are consistently enriched in the AO high green and red fluorescence region, although, for a given cell type, maximal enrichment is patient-dependent. Finally, cancer cell enrichment and lymphocyte exclusion can be done simultaneously. Cells from sputum were initially fixed, stained with AO, sorted on a dual parameter flow sorter, and classified into six groups corresponding to two ranges of green and three ranges of red fluorescence intensities. Cells of each region were stained by the method of Papanicolaou and differential counts were performed to determine the relative frequencies (i.e., purities) of leukocytes, macrophages, squamous cells, and cancer cells, in sorted and unsorted (i.e., control) samples. The average purity of leukocytes (81%), macrophages (6%), squamous cells (11%), and cancer cells (2%) varied markedly from sample to sample. However, the largest enrichment values (i.e., ratio of purity of a cell type in a sorted sample to its purity in the unsorted control sample) achieved for cancer cells consistently occurred for each patient sample in the region corresponding to high green and high red fluorescence intensities. Experimentally, a cancer cell average enrichment of sixteen-fold was obtained by this method. Additionally, fluorescence intensity ranges which increased the enrichment for macrophages by cell sorting typically excluded leukocytes and squamous cells, and vice versa. Finally, red fluorescence intensity was the primary discriminatory parameter for all cell types studied, although the additional use of green fluorescence intensity significantly increased cancer cell enrichment rates.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981661 TI - [Treatment with vitamin D, D hormones and synthetic analogs]. PMID- 2981662 TI - Ontogeny and characterization of basic somatomedin receptors in rat placenta. AB - We investigated the binding of [125I]basic somatomedin (B-SM) (insulin-like growth factor-I/SM-C in rat placenta. Low but specific [125I]B-SM binding to placenta was detected in rats from the end of the second week (14 days) of gestation. The binding increased until about 17 days of gestation and then reached a plateau. Binding characteristics did not alter during gestation. In contrast to previous observations [125I]B-SM binding to the tissue was competed more effectively by B-SM than multiplication stimulating activity (a rat counterpart of IGF-II). Insulin competed for the binding at a higher concentration. The Scatchard plot derived from the binding competition data was curvilinear. Autoradiograms of affinity labeled B-SM receptors in placentas at different gestational age subsequent to sodium dodecyl sulfate-gel (7.5%) electrophoresis revealed labeled constituents with approximate mol wt greater than 330 K and 140 K in the presence of reductant and greater than 330 K in the absence of reductant. Detailed analysis of the receptor in term placenta subsequent to 5-13% gradient gel containing sodium dodecyl sulfate revealed two more labeled components (mol wt, 250 K and 36 K) irrespective of the reductant. However, the intensity of the 250 K and 36K was low compared to 140 K and 330 K components. These results suggest that B-SM receptor in rat placenta, similar to human placenta, binds preferentially to type I or 140 K subunit of the 330 K oligomeric SM receptor. PMID- 2981664 TI - Parathyroid hormone modulation of 25-hydroxyvitamin D3 metabolism by cultured chick kidney cells is mimicked and enhanced by forskolin. AB - In order to determine whether cAMP mediates the effects of PTH on the metabolism of 25-hydroxyvitamin D3 (25-OH-D3) on chick kidney cells in primary culture, the effect of forskolin on the production of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] and 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3] was assessed. In 4-h incubations with [3H]25-OH-D3 and forskolin, (1-10 microM) [3H]1,25-(OH)2D3 accumulation was increased 50-100%, and that of [3H]24,25-(OH)2D3 was decreased 30-60%. PTH (1-10 ng/ml) brought about identical changes. Similar results were observed when cultures were preincubated with nonradioactive 25-OH-D3 for 4 h in the presence of PTH and forskolin, followed by a 30-min incubation with radioactive substrate. At a low concentration (0.05 microM), forskolin alone had no effect on the metabolism of [3H]25-OH-D3 but markedly enhanced that of PTH. At maximal concentrations of PTH (10 ng/ml) and forskolin (10 microM), the effects of the two on 25-OH-D3 metabolism were not additive. Both PTH and forskolin decreased the further metabolism of [3H]1,25-(OH)2D3, probably by inhibiting its 24 hydroxylation, but there are also cycloheximide-sensitive steps in the metabolism of 1,25-(OH)2D3 that are not affected by PTH and forskolin. In time course experiments, increased [3H]1,25-(OH)2D3 accumulation could be observed before the detection of 24-hydroxylase activity suggesting that the primary effect of PTH and forskolin is on the production of [3H] 1,25-(OH)2D3 rather than its catabolism. Raising the calcium concentration of the medium to 2.5 mM from the normal 1.8 mM or lowering it to 0.5 mM for 24 h in serum-free medium did not alter the response of 25-OH-D3 metabolism to these agents. The results of these studies indicate that the effects of PTH on the metabolism of 25-OH-D3 by chick kidney cells are mediated by cAMP, since they can be enhanced and mimicked by forskolin, that they are exerted at the level of both 1- and 24-hydroxylase activity, and that they are not dependent on the calcium concentration of the medium. PMID- 2981663 TI - A novel type of vasopressin receptor on anterior pituitary corticotrophs? AB - Suspensions of rat anterior pituitary cells were exposed to corticotropin releasing factor (CRF) (5 nM) and various neurohormones (0.002-1000 nM). CRF induced secretion of ACTH was doubled by 0.1 nM arginine vasopressin (AVP), 0.2 nM arginine vasotocin, 1 nM oxytocin, 10 nM angiotensin II, and 100 nM noradrenalin; vasoactive intestinal peptide had no effect at 0.2-200 nM. CRF potentiation by AVP was also observed at lower concentrations of CRF. Since AVP appeared to be the most potent modulator of CRF-induced ACTH secretion, potentiation was further tested with specific antidiuretic and oxytocic agonists. Potentiation was clearly related to pressor biological activity, less so to antidiuretic, and hardly at all to oxytocic activities. However, even at 200 nM, the antipressor antagonists dPTyr(Me)AVP and d(CH2)5Tyr(Me)AVP had no effect on potentiation by AVP. The lack of antagonism was partly due to the agonistic effects of the antagonists on the pituitary gland, an effect not observed within vascular tissue. The results thus suggest that anterior pituitary vasopressin receptors resemble, but are not identical to, V1 (pressor and hepatic), do not resemble the V2 (renal), and might be classified as V3 (pituitary) receptors. PMID- 2981665 TI - The involvement of collagenolysis in ovulation in the rat. AB - Collagenolytic activity in ovarian follicles was previously demonstrated by using synthetic peptides and reconstituted collagen fibers. However, attempts to demonstrate degradation of ovarian collagen and to correlate collagenase activity with ovulation were not successful. By administration of L-(5-3H) proline, we have labeled ovarian and follicular collagen and followed collagenolytic activity by separation of 3H-hydroxyproline (3H-Hyp) from acid hydrolyzates of ovarian tissue by HPLC. The level of ovarian and follicular 3H-Hyp decreased by about 40% on the afternoon of proestrus or after exogenous stimulation of ovulation by human CG (hCG), and this decrease was abolished by blocking the surge of gonadotropins with Nembutal. To verify that the observed reduction in 3H-Hyp was due to the action of a typical collagenase, the collagenous fraction was prepared from ovarian tissue and from preovulatory follicles before and after the ovulatory stimulus. The extracts were treated with trypsin (25 min, 25 C, 0.01 mg/ml) plasmin and p-amino-phenyl-mercuric acetate to fully activate the collagenase extracted along with collagen. Both, enzymatic and chemical activation of collagenase in vitro resulted in degradation of collagen. This degradation could be inhibited by cysteine and EDTA; both are classic inhibitors of mammalian collagenases. The activity of ovarian collagenase increased within 3 h after hCG-stimulation, peaked at 5-fold 6 h after hCG, and declined afterwards. Administration of cysteine (0.001-0.01 mmol) into the bursal cavity of proestrous rats blocked ovulation and breakdown of ovarian collagen in a dose-dependent manner. Cysteine effectively inhibited ovulation even when injected 7 h after the hCG stimulus. Inhibitors of arachidonic acid metabolism prevent ovulation. Indomethacin (inhibitor of cyclooxygenase) and nordihydroguaiaretic acid (inhibitor of lipoxygenase) blocked ovulation and inhibited hCG-induced ovarian collagenolysis. Collectively, these results corroborate the essential role of collagenolysis in follicular rupture in mammals. PMID- 2981666 TI - Biosynthesis of the epidermal growth factor receptor in cultured human cells. AB - A 160,000 mol wt precursor of the epidermal growth factor (EGF) receptor has been identified in human A-431 carcinoma cells and skin fibroblasts. The presence of one discrete precursor band indicates the presence of a slow processing step. We have determined that this slow processing step involves the conversion of high mannose N-linked oligosaccharides on the receptor precursor to primarily complex oligosaccharides on the mature form of the receptor. This is shown by 1) the presence of fucose, a characteristic terminal sugar of complex oligosaccharides, in only the mature receptor and by 2) the susceptibility of the precursor to digestion with endoglycosidase H, which cleaves high mannose N-linked oligosaccharides, but not complex oligosaccharides from glycoproteins. The precursor to mature receptor transition half-time is 1.7 h in A-431 cells. This long transition half-time causes an accumulation of approximately 7.2 X 10(5) precursor molecules per cell (approximately 12% of the total population of EGF receptors). The net quantity of mature EGF receptors, but not of receptor precursors, is reduced when EGF is added to the culture medium of A-431 cells. The presence of EGF in the growth medium also decreases electrophoretic migration (as a result of increased phosphate incorporation) of the mature receptor, but not that of the precursor. The EGF-insensitive state of the precursor is most likely due to its intracellular location. PMID- 2981668 TI - Effects of fetal hypophysectomy and the in vitro treatment by pituitary extracts on the maturation of cultured ovine fetal adrenal cells. AB - In the first series of experiments, the ability of cultured adrenal cells from 113-day-old ovine fetuses to produce both cAMP and corticosteroids in response to ACTH-(1-24) or to fetal (FPE) or newborn acidic pituitary extracts (NPE) was investigated daily. Basal cAMP output did not change during the culture period. When cells from 124-day-old ovine fetuses, or from 5-day-old lambs, were repeatedly stimulated (2 h/day for 4 days) by ACTH-(1-24) or by pituitary extracts, the cAMP responses increased with the same pattern. Outputs on day 4 were 7-fold higher than those on day 1 for ACTH-(1-24)-matured cells or for cells matured by FPE or NPE. The steroid output induced by ACTH-(1-24) or by FPE or NPE developed identically during the experiment to become, on day 4, more than 40 fold higher than on day 1. The response to ACTH-(1-24) on day 5 was also identical both in cAMP and corticosteroids whether the cells had been previously treated with ACTH-(1-24) or with FPE or NPE, In the second set of experiments, adrenal cells from 124-day-old-ovine fetuses either intact of hypophysectomized (Hx) at 118 days of gestation were cultured for 6 days in the absence or presence of ACTH-(1-24). ACTH-(1-24) treatment resulted in a development of cAMP and corticosteroid responses to the hormone which was slower for cells from Hx than cells from control fetuses during the first 3 days of culture. Likewise both cAMP and corticosteroids responses to ACTH-(1-24) of adrenal cells from Hx fetuses cultured for 1 to 3 days in the absence of ACTH were lower than those of cells from control fetuses cultured under the same conditions. These results demonstrate that the pituitary from 124-day-old ovine fetuses contains trophic (and steroidogenic) substances in a sufficient amount to allow in vitro adrenal maturation. Moreover, it appears that high mol wt forms of ACTH, which are most probably extracted by the method we used, did not prevent the in vitro development of the response to ACTH-(1-24). Finally, they show that removal of pituitary hormones in vivo resulted in a decreased potency of fetal adrenal cells to respond in vitro to ACTH-(1-24). PMID- 2981667 TI - Peripheral-type benzodiazepine receptors in endocrine organs: autoradiographic localization in rat pituitary, adrenal, and testis. AB - We have used [3H]Ro5-4864, a ligand selective for peripheral-type benzodiazepine receptors, to identify and localize peripheral-type benzodiazepine receptors in endocrine organs. Autoradiographic studies reveal an uniform distribution of [3H]Ro5-4864 binding sites within the anterior, intermediate, and posterior lobes of the pituitary gland, with highest concentrations present in the posterior pituitary. In rat adrenal gland, specific binding sites for [3H]Ro5-4864 are found only in the adrenal cortex, with highest density in the zona glomerulosa and significantly lower concentrations in the zona fasciculata and zona reticularis. [3H]Ro5-4864-associated silver grains in the testis are intensely localized over the interstitial tissue; low concentrations of silver grains are present over the epithelium of the seminiferous tubules but are absent from the tubular lumen. These studies demonstrate a differential and discrete localization of peripheral-type benzodiazepine receptors in rat pituitary, adrenal, and testis. PMID- 2981669 TI - In vitro spontaneous and adrenocorticotropin-dependent maturation of the steroidogenic pathway of ovine fetal adrenal cells. AB - Adrenal cells from intact or hypophysectomized fetuses and newborn lambs cultured for 6 days under several conditions were tested for their ability to metabolize [14C]pregnenolone (P5) (5 X 10(-5) M) during a 2-h period. Also, their ability to produce P5 was investigated. At the beginning of the culture, the metabolism of P5 by adrenal cells from hypophysectomized fetuses was lower than that of cells from intact fetuses which in turn was lower than that of cells from newborns. After 6 days in culture in the absence of ACTH-(1-24), the metabolism of P5 by cells from both intact and hypophysectomized fetuses increase, whereas that of cells from newborns slightly decreases. Continuous ACTH-(1-24) (10(-8) M) treatment produced a striking increase in the capacity to metabolize P5 by cells from both intact and hypophysectomized fetuses and the maintainance of this capacity of cells from newborns was demonstrated. At the beginning of the culture the main product of [14C]P5 was corticosterone with cells from intact and hypophysectomized fetuses, and cortisol with newborn cells. After 6 days in culture in ACTH-free medium more than 50% of P5 added appeared as progesterone in all cases. Addition of ACTH-(1-24) to the culture medium induced within 5 days, a striking increase in the activation of 3 beta-hydroxysteroid dehydrogenase isomerase, 17 alpha-, 21, and 11 beta-hydroxylases of adrenal cells from fetuses. However, the activity of 17 alpha-hydroxylase in adrenal cells from hypophysectomized fetuses was lower than that of cells from intact fetuses. ACTH (1-24) treatment maintained the enzymatic activity of cultured newborn adrenal cells, except for 11 beta-hydroxylase which declined to about 50% of its activity on day 0. The capacity of fetal adrenal cells to produce P5 after acute ACTH-(1 24) stimulation increased 30-fold when cells were cultured in the absence of ACTH and up to 110-fold when cells were treated with ACTH-(1-24) from day 1 on. These results, together with those reported previously, show that the spontaneous in vitro maturation of fetal adrenal cell steroidogenic pathway is related to an increased activity of the 3 beta-hydroxylase dehydrogenase-isomerase and an enhanced capacity to produce P5. Both steps were stimulated further by ACTH. In addition, the hormone is an absolute requirement for the development and/or the maintenance of 17 alpha-hydroxylase activity. PMID- 2981670 TI - Differences in the behavior of luteinizing hormones of various species at the rat gonadal cell receptor site. AB - The ability of different LH-like hormones, such as hCG, PMSG/equine (e) CG, ovine (o) LH, eLH, and rat (r) LH, to bind to and stimulate steroidogenesis in two types of rat gonadal cells was studied under the same experimental conditions. In both Leydig and granulosa cells, the maximal steroidogenic responses elicited by optimal doses of different LHs present during a 2-h incubation were comparable. However, if the cells were exposed to the different LHs for a brief period and then subjected to interference with hormone action by removing the unbound hormone from the medium by washing or adding specific antisera, differences were observed in the amount of steroid produced during subsequent incubation in hormone-free medium. Thus, in the case of hCG, either of these procedures carried out at 15 or 30 min of incubation had little inhibitory effect on the amount of steroid produced at 2 h, the latter being similar to that produced by cells incubated in the continued presence of hCG for 2 h. With eCG and rLH, the effect was dramatic, in that there was a total inhibition of subsequent steroidogenic response. In cells exposed to eLH and oLH, inhibition of subsequent steroidogenesis due to either removal of the free-hormone or addition of specific antisera at 15 or 30 min was only partial. Although all of the antisera used were equally effective in inhibiting the steroidogenic response to respective gonadotropins when added along with hormones at the beginning of incubation, differences were observed in the degree of inhibition of this response when the same antisera were added at later times of incubation. Thus, when antisera were added 60 min after the hormone, the inhibition of steroidogenesis was total (100%) for eCG, partial (10-40%) for eLH and oLH, and totally lacking in cells treated with hCG. From this, it appears that hCG bound to the receptor probably becomes unavailable for binding to its antibody with time, while in the case of eCG and other LHs used, the antibody can still inhibit the biological activity of the hormone. Studies with 125I-labeled hormones further supported the conclusion that hCG differs from all other LHs in being most tightly bound and, hence, least dissociable, while eCG and rLH dissociate most readily; oLH and eLH can be placed in between these hormones in the extent of their dissociability. PMID- 2981671 TI - Evidence for multifactor regulation of the adrenocorticotropin secretory response to hemodynamic stimuli. AB - We have examined the contributions of corticotropin-releasing factor (CRF), arginine vasopressin (AVP), epinephrine, and oxytocin to the ACTH secretory responses to hemorrhage. The relative significance of each of these putative ACTH regulatory factors is undefined with respect to net ACTH secretion. Initially, the effects of selective systemic pharmacological blockade of individual factors on the ACTH response were examined. Immunoneutralization of CRF reduced resting ACTH levels below the detection limits of our RIA and abolished the secretory response to hemorrhage. Ganglionic blockade or treatment with a potent AVP antagonist reduced the ACTH secretory response by 55% and 38%, respectively. Further evidence for multifactor regulation of hemodynamically evoked alterations in ACTH secretion was obtained by measurement of the concentrations of these factors in the hypophysial portal circulation during hemorrhage. Immunoreactive CRF, AVP, oxytocin and epinephrine were present in the portal plasma at concentrations within a range shown to evoke ACTH secretion from cultured pituitary cells when presented alone or in combination. The concentrations of all of these were significantly elevated during hemorrhage. During atrial pulsation, a stimulus mimicking volume loading and associated with a reduction of systemic ACTH levels, we observed a significant decline in portal concentrations of immunoreactive AVP coupled with a nonsignificant trend toward reduced portal immunoreactive CRF levels. These observations are highly suggestive of multifactor regulatory control of ACTH secretion in response to hemodynamic stimuli. PMID- 2981672 TI - Thiocyanate feeding with low iodine diet causes chronic iodine retention in thyroids of mice. AB - Effects of KSCN on thyroidal iodine metabolism were studied in weanling mice fed a low iodide diet (LID) labeled continuously with 125I as iodide. The addition of KSCN (0.3 and 0.6 mg/g diet) resulted in the accumulation of an unusual iodinated protein within the follicles of the mouse thyroids. After 60 days, total thyroidal iodine was 4 times greater than in controls without thiocyanate. The iodinated protein was essentially insoluble at pH 8.0 and was very slowly released from the thyroids; it contained more MIT than DIT and little thyroid hormone. By use of three isotopes (125I, 127I, and 131I) and auto-radiographs, there were shown different iodinated proteins synthesized during high and low iodine intakes and coexistent but segregated in the colloid. Low doses of perchlorate or iodide inhibited or prevented accumulation of the essentially insoluble iodinated protein. It is suggested that when mouse thyroids are iodine depleted, thiocyanate increases the formation of an essentially insoluble iodinated thyroglobulin within the thyroid. Only a small fraction of the iodination may have occurred by this route, but the rate of formation exceeded the rate of release, so the product continuously accumulated. PMID- 2981673 TI - Specific receptors for vasopressin in the pituitary gland: evidence for down regulation and desensitization to adrenocorticotropin-releasing factors. AB - Specificity of binding of 3H-labeled arginine vasopressin [( 3H]AVP), down regulation of receptors, and desensitization were studied in anterior pituitary glands of both Wistar and Brattleboro rats. Studies using both crude membrane fractions and isolated cells of anterior pituitaries revealed the presence of a single population of binding sites with a Kd of approximately 1 nM. The receptor recognized the following peptides, with AVP = lysine vasopressin = vasotocin greater than oxytocin = 1-deamino-(8-D-AVP) greater than d-(CH2)5-Tyr-(Me)-Val4 AVP greater than 1-deaminopenicillamine-(Val4-D-Arg8)VP. Neither corticotropin releasing factor (CRF) nor any of the neuropeptides tested, including AVP ring and tail fragments, competed for tracer binding. Increased extracellular vasopressin levels due to chronic injections or long term adrenalectomy decreased receptor density by 80%, while oxytocin was less effective than AVP. Comparing binding data in Brattleboro homozygotes and heterozygotes revealed that AVP levels within the physiological range could down-regulate pituitary receptors as well. This could not be caused by occupation of sites by endogeneous vasopressin, since injection of large doses of peptide decreased tracer binding by less than 10%. Loss of pituitary receptors reduced 1) enhancement by AVP of CRF-induced cAMP accumulation, 2) intrinsic CRF-like activity and 3) synergistic effect of AVP on ACTH secretion elicited by CRF. This study thus provides evidence for the presence of highly specific vasopressin receptors in the anterior pituitary, which may undergo homologous down-regulation and desensitization in terms of cAMP production and ACTH release. PMID- 2981674 TI - Human pancreatic growth hormone-releasing factor-44 differentially stimulates release of stored and newly synthesized rat growth hormone in vitro. AB - Effects of synthetic human pancreatic GH-releasing factor-44 (hpGRF-44) on synthesis and release of rat pituitary GH and PRL were examined in vitro in a static incubation system. A double label, specific immunoprecipitation protocol permitted simultaneous study of hormone synthesis as well as release of both stored and newly synthesized hormone. Synthetic hpGRF-44 (0.3 and 3.0 nM) stimulated the release of stored GH 240% beyond the basal level, while simultaneously stimulating the release of newly synthesized GH by 610%. Despite the stimulation of release, hpGRF-44 did not alter GH synthesis (102% of control value). A small but statistically significant increase in release of stored PRL occurred in response to hpGRF-44, while release of newly synthesized PRL and PRL synthesis were unaffected. In contrast, 1 mM (Bu)2cAMP stimulated the release of both newly synthesized and stored GH and PRL. We conclude that hpGRF-44 differentially stimulates GH release from separate intracellular compartments and that the lactotroph may also, under certain conditions, respond to this secretagogue. PMID- 2981675 TI - Involvement of plasma membrane enzymes in the proteolytic cleavage of luteinizing hormone receptor. AB - In vitro degradation of LH receptor after occupancy by hCG was studied. Rat ovarian membranes labeled with [125I]iodo-hCG were incubated at 37 C; as a result, 30-40% of the radioactivity initially bound was rendered soluble in the medium. The molecular complexes in the medium and in incubated membranes solubilized with 1% Triton X-100 were then cross-linked with glutaraldehyde and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Particulate receptor-[125I]iodo-hCG complex exhibiting an apparent mol wt of 125,000 was cleaved during incubation into two distinct components (mol wt, 96,000 and 74,000) which appeared in the medium. Using tritiated hCG (beta-subunit labeled) instead of radioiodinated hCG (alpha-subunit labeled), these same two components were also observed, indicating that they both contain intact hCG (alpha and beta) as a part of their structure. In addition to the hormone (mol wt, 48,000), these two components contain receptor fragments with mol wt of 64,000 or 38,000, demonstrated directly by labeling the particulate receptor itself with periodate-tritiated borohydride before tagging with unlabeled hCG and in vitro incubation. These receptor fragments were purified from the medium by hCG-directed immunoaffinity chromatography and detached from the hormone by pH treatment. The intact receptor extracted from the membranes with detergent and purified identically in the absence of proteolysis migrated as a 90,000 mol wt polypeptide. These results demonstrate that after hormone occupancy, proteolytic cleavage of the 90,000 mol wt receptor polypeptide occurs at two specific sites. Thiol-blocking agents selectively prevented the appearance of the larger component (hCG coupled to 64,000 mol wt receptor fragment), while metal-chelating agents markedly decreased the appearance of the smaller component (hCG coupled to 38,000 mol wt receptor fragment) in the medium. Identical observations, obtained upon incubation of plasma membranes purified by sucrose density gradient centrifugation, suggest that plasma membrane enzymes are involved. PMID- 2981676 TI - CI628 inhibits follicle-stimulating hormone (FSH)-induced increases in FSH receptors of the rat ovary: requirement of estradiol for FSH action. AB - Although estradiol (E2) alone does not increase receptors for FSH in granulosa cells, E2 priming before administration of FSH increases numbers of FSH receptors significantly compared with FSH alone. We hypothesized that if E2 is required for FSH to increase its own receptor, blocking estrogen action should prevent FSH induced increases in FSH receptors. Five groups of hypophysectomized rats were injected sc with: saline at 0 h; the antiestrogen CI628 (1 mg) at -6 h; human FSH (hFSH, 2 micrograms) at 0 h; CI628 at -6 h, then hFSH at 0 h; and CI628 plus E2 (2 mg) at -6 h, then FSH at 0 h. Animals were decapitated at 0, 6, 12, or 24 h, and granulosa membrane receptors for FSH, LH, and nuclear receptors for E2 were measured. LH receptor levels increased only after administration of E2 before hFSH. Treatment with hFSH for 6 h increased numbers of FSH receptors 3-fold (P less than 0.01) without any increase in numbers of E2 receptors. At 12 and 24 h, hFSH increased numbers of FSH and E2 receptors 6- and 7-fold (P less than 0.01) over controls. CI628 prevented the hFSH-induced increases in FSH receptors at 6, 12, and 24 h. Administration of E2 concomitant with CI628 before hFSH significantly reversed the inhibitory effects of CI628 on hFSH-induced increases in FSH receptors. There were no changes in affinity of FSH or E2 receptors from 0 to 24 h. To determine whether E2 was acting on the adenylate cyclase system, the ability of hFSH to increase the content of cAMP in granulosa cells in each treatment group was determined. After an iv injection of hFSH, cAMP levels were similar in CI628- and saline-treated rats but had increased 6-fold (P less than 0.01) in hFSH or CI628 plus hFSH-treated animals. Thus, blocking hFSH-induced increases in FSH and E2 receptor appeared to have no effect on FSH stimulation of cAMP. In conclusion E2 appears to be required for FSH action, perhaps by acting within granulosa cells distal to the cAMP-adenylate cyclase system. PMID- 2981677 TI - Inhibition of corticotropin release during hypothermia: the role of corticotropin releasing factor, vasopressin, and oxytocin. AB - To investigate the mechanism by which ACTH secretion is inhibited during hypothermia, hypophysial portal blood was collected from euthermic and hypothermic rats, and the concentrations of corticotropin-releasing factor (CRF), vasopressin (AVP), and oxytocin (OT) were measured by RIA. Whereas CRF levels in portal plasma were not different in the two groups, AVP and OT levels were significantly lower in hypothermic rats. The concentration of AVP and OT in peripheral plasma was also significantly lower in hypothermic rats compared with euthermic controls. The pituitary responsiveness to CRF during hypothermia was tested in vivo and in vitro. In pentobarbital-anesthetized male rats injected iv with 0.1 or 1.0 nmol CRF, the ACTH response was significantly smaller in hypothermic compared with euthermic animals. However, hemipituitaries superfused at 31 C released the same amount of ACTH in response to 1 nM CRF as hemipituitaries superfused at 37 C (31 C, 541 +/- 90 pg; 37 C, 563 +/- 29 pg) despite reduced baseline secretion (31 C, 77 +/- 10 pg/10 min; 37 C, 114 +/- 14 pg/10 min; P less than 0.05). The data suggest that the inhibition of ACTH secretion during hypothermia is mediated by decreased hypothalamic secretion of AVP and OT which in turn decreases the pituitary responsiveness to CRF. PMID- 2981678 TI - 12-O-tetradecanoyl phorbol-13-acetate stimulates rat growth hormone (GH) release through different pathways from that of human pancreatic GH-releasing factor. AB - The mechanism(s) of action of 12-O-tetradecanoyl phorbol-13-acetate (TPA) on rat (r) GH release was studied in primary rat pituitary cell cultures. TPA stimulated rGH release (3.2- to 4.1-fold above control value) and rTSH and rLH release (1.4- and 1.7-fold above control values, respectively), but not rPRL release. The ED50 of TPA on rGH secretion was 1.3 X 10(-9) M compared to 4.5 X 10(-11) M for human pancreatic GH-releasing factor [hpGRF-(1-44)]. If maximally effective doses of TPA or hpGRF-(1-44) were added to the cells, the magnitudes of the increase in rGH release were quite similar for both agents when the incubation period was less than 12 h. When (Bu)2cAMP was added simultaneously with various doses of TPA, (Bu)2cAMP increased rGH release beyond the maximal effect of TPA. There was an additive effect when hpGRF-(1-44) and TPA were used to stimulate rGH release. These results indicate that TPA enhances rGH release through a different pathway than hpGRF-(1-44). TPA failed to increase the formation of intra- and extracellular cAMP, whereas hpGRF-(1-44) increased both, suggesting that TPA stimulates rGH release through an cAMP-independent pathway(s). Protein kinase C has been postulated to be a receptor for TPA in human platelets. When phospholipase C, which activates protein kinase C via the formation of diacylglycerol, was added to the cells, rGH release was stimulated in a dose dependent manner. This effect was not blocked by indomethacin. These results may suggest that activation of protein kinase C leads to rGH release. The observations are consistent with the hypothesis that TPA activates protein kinase C and causes the release of rGH in normal pituitary cells in culture. These findings indicate that the mechanism(s) of action of TPA on rGH release is different from that of hpGRF-(1-44). PMID- 2981680 TI - Intestinal absorption of arsenate in the chick. AB - The intestinal absorption of arsenate(As(V)) has been investigated in the chick by means of the in situ ligated duodenal loop technique. By this procedure, it was observed that arsenate is rapidly and essentially completely absorbed (80 95%) from the lumen at As(V) concentrations up to 5 mM, declining to about 50% absorption at 50 mM. Transfer from the intestinal lumen to the mucosal cells at low As(V) concentration (0.1 mM) is rapid, while transfer from the mucosal cells to the body occurs more slowly. At stable As(V) concentrations greater than 1 mM, fractional mucosal cell accumulation of As(V) remains constant, while fractional transfer to the body declines. However, total mucosal accumulation of As(V) and that transferred to the body increase in a linear logarithmic fashion from 0.05 to 5 mm As(V). The results indicate that As(V) readily penetrates both the mucosal and serosal surfaces of the epithelial membrane. Furthermore, arsenate and phosphate do not appear to share a common transport pathway in the duodenum and no evidence was obtained for any interaction between the two at this level. Vitamin D3 administration to rachitic chicks was effective in significantly elevating duodenal arsenate absorption, acting primarily to enhance serosal transport. PMID- 2981679 TI - Daily administration of melatonin delays rat vaginal opening and disrupts the first estrous cycles: evidence that these effects are synchronized by the onset of light. AB - The effect of daily melatonin administration was investigated in the immature female rat. Starting on day 15 of age, 100 micrograms melatonin were injected sc at different times of the day in animals housed in 12 h of light, 12 h of darkness or 16 h of light, 8 h of darkness. Melatonin given 9-11 h after the onset of light in both lighting regimens resulted in a 10-day delay of vaginal opening, a dissociation of the relation between vaginal opening and first proestrus, and a disruption of the initial estrous cycles. The same dose of melatonin given at other times during the photoperiod had no effect on sexual maturation. GnRH secretion in melatonin-treated animals was decreased, as judged by 30% lower pituitary GnRH receptor number in animals killed after opening of the vagina. During the diestrous phases, plasma levels of LH, FSH, and 17 beta estradiol were similar to those in control rats, but during proestrus, the surge of FSH was higher, and the peak of estradiol was higher and of a longer duration. This hormonal pattern suggests a build-up of hormones in secreting cells, which follows the lower incidence of proestrous phases in melatonin-treated rats. This build-up of FSH was indeed present, with higher concentrations in the pituitary during diestrus after melatonin treatment, while pituitaries removed during proestrus had lower contents of FSH. These results confirm that chronic melatonin administration delays sexual maturation of female rat, probably by retarding maturation of hypothalamic GnRH-producing cells. Thus, melatonin could modify basal GnRH secretion or pulsatile release. Pituitary and ovarian responsiveness do not seem to be affected, since proestrous surges of 17 beta-estradiol, LH, and FSH occur, albeit at a reduced frequency. The results also show that there is a window of maximum sensitivity to administration of melatonin 9-11 h after the onset of light, and that this window of sensitivity is synchronized by the onset of light. This raises the possibility that the abnormal presence of endogenous melatonin during this period of the day could induce abnormal sexual development. PMID- 2981682 TI - Isolation and characterization of the Fnr protein, the transcriptional regulator of anaerobic electron transport in Escherichia coli. AB - The Fnr protein, the transcriptional regulator of the expression of anaerobic respiratory functions in Escherichia coli, was purified to homogeneity from soluble extracts of a strain harbouring the fnr gene in an expression vector. The identity of the isolated protein was confirmed by comparing its amino-terminal sequence with that predicted from nucleotide sequence of the fnr gene. It appeared that eight amino-terminal amino acids had been removed post translationally from the bulk of the isolated Fnr protein. The molecular mass of the isolated protein (Mr 28 000) was consistent with a monomeric state, but sedimentation coefficients for the cellular (4.1 S) and the isolated (2.9 S) Fnr protein suggest that it may exist as a dimer in the bacterial cells. The Fnr protein bound DNA. However, the binding activity was not specific for the regulatory regions of relevant genes and it could not be stimulated by a variety of conditions or potential effectors. Two of the four cysteine residues of the Fnr protein were alkylated by iodoacetic acid and this could have functional significance in rendering the protein redox-sensitive. PMID- 2981681 TI - DNA regions associated with the nuclear matrix of Ehrlich ascites cells expose single-stranded sites after deproteinization. AB - Ehrlich ascites cells were pulse-labeled with [3H]thymidine and subjected to prolonged labeling with [14C]thymidine. The isolated nuclei were digested with the restriction endonuclease BspRI and then processed to yield a 'matrix fraction' and a 'non-matrix fraction'. The DNA fragments purified from these fractions and from whole digested nuclei were examined for nitrocellulose-binding sites before and after digestion with single-strand-specific (S1) nuclease. Both, pulse-labeled and long-time-labeled fragments, isolated from the matrix fraction, exhibited a significantly increased content of nitrocellulose-binding sites. The major portion of these sites were rendered non-binding by digestion with single strand-specific nuclease and consisted most probably of structures exposing relatively small stretches of non-base-paired DNA. The nature of the minor portion of binding sites which was insensitive to single-strand-specific nuclease is not clear. Both types of binding sites are possible candidates for mediating the attachment of DNA to the nuclear matrix. PMID- 2981683 TI - Structure of the cytochrome c oxidase complex of rat liver. 1. Studies on nearest neighbour relationship of polypeptides with cross-linking reagents. AB - Isolated rat liver cytochrome c oxidase was cross-linked with the cleavable reagents dimethyl-3,3'-dithiobispropionimidate (DTBP), 3,3' dithiobis(succinimidyl)propionate (DSP) and cupric di(1,10-phenanthroline) (CuP). The cross-linked products were separated by high-resolving two-dimensional dodecyl sulfate gel electrophoresis, which separates all thirteen polypeptides of the mammalian enzyme. With cupric di(1,10-phenanthroline) seven polypeptides (I III, Va, Vb, VIIb and VIII) were cross-linked with each other and with itself, indicating the occurrence of free -SH groups in these polypeptides and a rearrangement of the native structure of the complex by cupric di(1,10 phenanthroline). With dimethyl-3,3'-dithiobispropionimidate or 3,3' dithiobis(succinimidyl)propionate all nuclear-coded polypeptides, with the exception of polypeptide VIIa, formed cross-linked products with the three 'catalytic' polypeptides I-III, which are coded on mitochondrial DNA. Five additional cross-linked pairs were found between nuclear coded polypeptides. The close arrangement of nuclear coded polypeptides with the catalytic polypeptides suggests a regulatory function of these polypeptides. PMID- 2981684 TI - Structure of the cytochrome c oxidase complex of rat liver. 2. Topological orientation of polypeptides in the membrane as studied by proteolytic digestion and immunoblotting. AB - The orientation of the thirteen polypeptides of rat-liver cytochrome c oxidase in the inner mitochondrial membrane was studied by proteolytic digestion of mitoplasts and sonicated particles. After separation by sodium dodecylsulfate gel electrophoresis proteins were transferred on nitrocellulose, and individual polypeptides were identified by incubation with polypeptide-specific antisera, followed by fluorescein-isothiocyanate-conjugated protein A. The three catalytic polypeptides I-III and seven nuclear coded polypeptides (IV, Vb, VIa, VIc, VIIa, VIIb and VIII) were found accessible to proteases from the cytoplasmic phase. Polypeptides II, IV, Va, Vb and VIa were accessible from the matrix phase, indicating a transmembraneous orientation of polypeptides II, IV, Vb and VIa. Together with data on cross-linking and on cytochrome-c-protected labeling of polypeptides, a model of the cytochrome c oxidase complex was developed. It is suggested that the cytochrome c binding site on polypeptide II is surrounded by several nuclear-coded polypeptides, which may modulate the affinity of the enzyme towards cytochrome c. PMID- 2981685 TI - N-Bromoacetyl-amino-cyanopindolol: a highly potent beta-adrenergic affinity label blocks irreversibly a non-protein component tightly associated with the receptor. AB - A new chemical affinity label for the beta-adrenergic receptor, based on the structure of pindolol, has been synthesized and iodinated with 125I. The compound, N-bromoacetylamino-cyanopindolol (BAM-CYP), has an apparent dissociation constant of 44 +/- 7 pM towards the turkey erythrocyte membranes. This compound blocks irreversibly both the ability of beta-adrenergic receptors to bind 125I-cyanopindolol and the ability of beta-receptors to activate adenylate cyclase in the presence of beta-agonists. Furthermore, the irreversible binding of BAM-CYP to half of the beta-receptor sites abolishes the ligand binding activity of all the sites. These findings suggest that the beta-receptor is oligomeric in its native state. Although 125I-BAM-CYP blocks irreversibly and specifically the beta-adrenergic receptor, it does so by labeling a non-protein component, most probably a water-soluble lipid. The labeling is stereospecific since it is prevented by l-propranolol and not by d-propranolol. It is suggested that this lipid is tightly associated with the receptor in close proximity to the binding site. It is also suggested that this water-soluble lipid fraction may prove crucial for the optimal interaction between the beta-adrenergic receptor and the components of adenylate cyclase. PMID- 2981686 TI - Sequential ADP-ribosylation pattern of nucleosomal histones. ADP-ribosylation of nucleosomal histones. AB - The pattern of nucleosomal histones poly(ADP-ribosyl)ation is changed under conditions which affect the poly(ADP-ribosyl)ation state of the enzyme. At low NAD concentrations the enzyme can poly(ADP-ribosyl)ate histones H1 and H1, H2A, A2A, and H2B. However at NAD concentrations above 10 microM the enzyme preferentially poly(ADP-ribosyl)ates histone H1 to a hyper ADP-ribosylated form. Furthermore we have observed hyper ADP-ribosylation of histone H2B at NAD concentrations of 10 microM suggesting that histone H2B can undergo the same type of ADP-ribosylation pattern as histone H1. Also at higher NAD concentrations an elongation of the polymer attached to the enzyme and other nuclear proteins takes place. PMID- 2981687 TI - A calcium-dependent protease associated with the neural cytoskeleton. Purification and partial characterisation. AB - Calcium-dependent protease activity was found associated with a neurofilament enriched cytoskeleton isolated from the bovine spinal cord. The protease was extracted from the cytoskeleton by 0.6 M KCl, and purified to apparent homogeneity (3300-fold) by chromatography on organomercurial-Sepharose 4B, casein Sepharose 4B, and Sepharose CL-6B. A cytosolic calcium-dependent protease was similarly purified from the bovine spinal cord, after the cytosol was fractionated on DEAE-cellulose. Both cytoskeleton-bound and cytosolic enzymes had an apparent molecular mass of 100 kDa as judged by gel filtration, and consisted of two subunits (79 kDa and 20 kDa) upon sodium dodecyl sulfate/polyacrylamide gel electrophoresis. Both enzymes exhibited caseinolytic activity with 0.5 mM Ca2+ and above, and the activity was strongly inhibited by various thiol protease inhibitors. In the presence of 0.1-0.2 mM Ca2+, the 68-kDa and 160-kDa components, and to a lesser extent the 200-kDa component, of the neurofilament triplet polypeptides were degraded by the cytosolic protease, whereas the cytoskeleton-bound protease needed two-fold higher concentration of Ca2+ to degrade the neurofilaments. Nevertheless, the cytoskeleton-bound protease in situ, i.e. before its extraction form the cytoskeleton by 0.6 M KCl, preferentially degraded the 160-kDa component in the presence of 0.1-0.2 mM Ca2+, suggesting that a proper locational relation of this enzyme to the neurofilament structure is a prerequisite to its preference for the 160-kDa component. It appears that a factor or factors involved in such an interaction between the protease and the neurofilament were eliminated during the course of enzyme purification. The glial fibrillary acidic protein was almost insensitive to the proteases purified in the present study. PMID- 2981688 TI - Direct assessment of beta-adrenergic receptors in intact rat adipocytes by binding of [3H]CGP 12177. Evidence for agonist high-affinity binding complex and for beta 1 and beta 2 receptor subtypes. AB - The characteristics of the binding of the hydrophilic beta-adrenergic antagonist [3H]CGP 12177 to intact rat adipocytes were studied at 37 degrees C and 6 degrees C. At both temperatures and at 90% saturation, the non-specific binding was less than 30% of the total binding. At 37 degrees C, specific [3H]CGP 12177 binding was rapid, reversible of high affinity (1.8 +/- 0.4 nM) and saturable. The number of specific binding sites per adipocyte increased with the fat cell size (about 35 000 and 115 000 sites per cell in adipocytes with diameters of 60 microns and 88 microns, respectively) but remained constant when expressed per unit fat cell surface area. Displacement of [3H]CGP 12177 bound to adipocytes by unselective and selective beta-antagonists was stereospecific, had the same characteristics as those found in adipocyte membranes and showed a heterogeneous specificity for beta 1 and beta 2 adrenergic subtypes. In contrast, beta agonist competition curves, which modeled to two affinity-states of binding, showed high-affinity state Kd values for agonists 10-25-times higher than those found in membranes under the same experimental conditions. At 6 degrees C, although the number and affinity of the specific binding sites for [3H]CGP 12177 were the same as those found at 37 degrees C, the Kd value for (-)-isoproterenol binding to the high affinity state of these sites (3.0 +/- 0.5 nM) was 25-times lower than at 37 degrees C and similar to the value found in membrane preparations (1.5-4 nM). These results show that the [3H]CGP 12177 specific binding sites detected on intact adipocytes represent the physiological beta-adrenergic receptors. Moreover, this study extends to the adipocyte the validity of the model recently proposed for other cell lines, according to which in intact cells, but not in membranes, agonist-binding promotes a rapid and temperature-dependent conformational change of the beta-adrenergic receptors, leading to a progressive loss of capacity of agonists to form a high-affinity complex. PMID- 2981689 TI - Effects of denaturation and methylation on the degradation of proteins in cultured hepatoma cells and in reticulocyte cell-free systems. AB - Radioiodinated, native and denatured bovine serum albumin (albumin) beta lactoglobulin and cytochrome c were introduced into hepatoma tissue culture cells by erythrocyte-ghost-mediated microinjection, and their rates of degradation were compared. Denatured albumin was degraded at 20% of the rate of undenatured albumin, denatured beta-lactoglobulin was degraded three times faster than undenatured beta-lactoglobulin, while denatured and undenatured cytochrome c were degraded at the same rate. Thus, denaturation does not affect the rates of intracellular breakdown of microinjected proteins in a simple predictable way. Exhaustive methylation did not inhibit the degradation of denatured beta lactoglobulin or albumin, indicating that, like their undenatured counterparts, they are not degraded via the ubiquitin pathway. In reticulocyte lysates, in the presence of ATP, denatured albumin and beta-lactoglobulin were broken down at slightly slower rates than the parent proteins. Exhaustive methylation of both denatured and undenatured proteins completely abolished their ATP-dependent breakdown. This inhibition is consistent with the hypothesis that free -NH2 groups are required for the attachment of ubiquitin prior to degradation in this system. Removal of an ammonium sulfate fraction from reticulocyte lysates produces a proteolytic system markedly different from the whole lysate [Speiser, S. & Etlinger, J. D. (1983) Proc. Natl Acad. Sci. USA 80, 3577-3580]. In this system both denatured and undenatured albumin and beta-lactoglobulin were degraded essentially independently of ATP. Methylation only slightly decreased the breakdown of denatured proteins, suggesting that they are not degraded via the ubiquitin pathway. A possible explanation of these results is that removal of the ammonium sulfate fraction unmasks an ATP-independent proteolytic system unrelated to the ubiquitin pathway. PMID- 2981690 TI - Regulation of thymocyte proliferative response by macrophage-derived prostaglandin E2 and interleukin 1. AB - The antagonistic effects of macrophage (M phi)-derived interleukin 1 (IL 1) and prostaglandin E2 (PGE2) on thymocyte proliferation were uncoupled and studied. Elimination of PGE2 from the culture medium of prostaglandin E2-stimulated M phi was achieved by dialysis of the media or by indomethacin treatment of the M phi. IL1 secretion appears to be PGE2 independent. Experiments using exogenous PGE2 revealed a quantitative relationship between the two monokines. PGE2 (1.25 ng/ml) reduced the proliferative effect of IL 1 produced by 1.5 X 10(5) peritoneal M phi to 50%. This PGE2 dose increased significantly the levels of intracellular cAMP. Separation of peritoneal exudate M phi on a bovine serum albumin discontinuous gradient demonstrated that the main part of PGE2 synthesis was in a fraction of lower density, large M phi, whereas IL 1 activity was detected in all tested fractions. PMID- 2981691 TI - The orientation of nucleus, nucleus-associated body and protruding nucleolus in aggregating Dictyostelium discoideum. AB - Dictyostelium discoideum growing or developing on cellulose dialysis membranes were fixed with acrolein vapour for electron microscopy. In interphase amoebae, nucleoli began to protrude from the nuclei. The percentage of cells with protruding nucleoli increased during aggregation by a value approximately twice as high in aggregation streams as in centers. Cells in pseudoplasmodia showed only a low percentage and protrusions disappeared at early culmination stage. The protrusions did not reappear when cells from dissociated pseudoplasmodia migrated toward cAMP. Thus the formation of the protrusions did not depend solely on chemotaxis; rather, it was specific to the aggregation stage. In aggregation streams, the nucleus was anterior in the cell, with the protrusion at its anterior periphery. In contrast, the nucleus associated body (NAB) was evident at the cell's mid-point. This orientation of nucleus and NAB in the aggregating slime mould amoeba is contrary to that seen in human neutrophils or cultured mouse 3T3 cells. PMID- 2981692 TI - Production of latent collagenase by human umbilical vein endothelial cells in response to angiogenic preparations. AB - Three preparations known to be angiogenic in vivo and which stimulate production of latent collagenase by cultured bovine capillary endothelial (BCE) cells were tested for their ability to stimulate production of latent collagenase by cultured human umbilical vein endothelial (HUVE) cells. Bovine retinal extract and murine adipocyte-conditioned medium had no effect on production of latent collagenase by HUVE cells at concentrations that were effective in stimulating production of latent collagenase by BCE cells. However, with higher concentrations of bovine retinal extract, production of latent collagenase by HUVE cells was stimulated. Human hepatoma cell sonicate stimulated production of latent collagenase by HUVE cells in a dose-dependent manner. The concentration of human hepatoma cell sonicate which stimulated production of latent collagenase by HUVE cells was lower than the concentration that was effective for the stimulation of production of latent collagenase by BCE cells. Plasminogen activator production by HUVE cells was unaffected by human hepatoma cell sonicate. Varying the concentration of serum in HUVE cultures did not affect the stimulation of latent collagenase production by human hepatoma cell sonicate, suggesting that serum components neither block nor stimulate the action of the collagenase-inducing factor. Although human hepatoma cell sonicate is reported to stimulate endothelial cell multiplication, purified and partially purified endothelial cell mitogens had no effect on production of latent collagenase. Thus, at least two preparations which contain angiogenic activity will stimulate production of latent collagenase by HUVE cells. PMID- 2981693 TI - Relationship of ultraviolet light-induced DNA-protein cross-linkage to chromatin structure. AB - The production of banding patterns in metaphase chromosomes by restriction enzymes is inhibited by ultraviolet (UV) irradiation. Irradiation of fixed chromatin produces a 15-fold decrease in DNA extraction by restriction enzymes in comparison with that observed by irradiation before fixation. Alcohol-acid fixation of chromatin produces two major changes, the extraction of histones and dehydration. The effect of UV light is probably the result of a net increase in the yield of DNA-protein cross-links at comparable fluences of UV light and of the stabilization of the structural changes in the fixed chromatin fibril induced by the photoadducts. The X-irradiation of cells before fixation, as well as the rehydration of fixed chromatin, increases the extraction of DNA from fixed chromatin irradiated with UV light to levels similar to or even higher than those obtained with living cells. The effect of UV light before and after fixation on the extraction of DNA by restriction enzymes and proteinase K can be related to changes in chromatin structure and DNA conformation. PMID- 2981695 TI - A method to measure receptor binding of ligands with low affinity. Application to plasma proteins binding assay with hemopoietic cells. AB - A gradient has been developed for separating free ligand from ligand bound to cells growing in suspension. This method can be used with all kinds of ligand but it is particularly useful for those ligands having the tiresome tendency to adhere to the cells non-specifically or to polymerize by themselves. This is the case of fibronectin, fibrinogen, immunoglobulins and many other plasma proteins. The gradient consists of two layers: an upper aqueous phase and a lower hydrophobic organic phase. The aqueous phase, a sucrose buffer, allows the cells to become well dispersed before they enter the hydrophobic phase which excludes the free ligand efficiently. This reduces non-specific binding and allows the accurate measurement of specific binding which could not be obtained with a gradient made of a single phase. Depending upon the size and the density of the cells, and the nature of the ligand, the assay method can be modified by changing the density or the nature of the hydrophilic and hydrophobic phases. PMID- 2981694 TI - Characterization of ouabain-resistant mutants of the preadipocyte Ob17 clonal line. Adipose conversion in vitro and in vivo. AB - After exposure to a mutagenic drug (ethylmethanesulfonate), mutant clones of Ob17 preadipose cells resistant to the cytotoxic action of ouabain have been isolated. Their ability to grow in serum-supplemented medium containing greater than or equal to 3 mM ouabain is consistent with the decreased ouabain sensitivity of K+ transport observed in one of these clones (Ob17-OR11). Ouabain-resistant mutants retain their ability to convert into adipose-like cells in vitro with a frequency similar to that of original Ob17 cells. Long-term maintenance of Ob17-OR11 cells in ouabain-enriched medium leads to a higher frequency of adipose conversion, which is linked to a rather specific and limited growth-promoting effect of the drug. Undifferentiated Ob17-OR11 cells, when injected subcutaneously into athymic mouse, give rise in vivo to a fat pad containing ouabain-resistant mature adipocytes. Therefore, Ob17-OR11 clone should be a useful tool to distinguish in vivo between intrinsic and extrinsic factors involved in the differentiation of adipose precursor cells. PMID- 2981696 TI - Neurotoxicity of Adriamycin and misonidazole in the mouse. AB - The neurotoxicity of the anticancer drug adriamycin was investigated in the peripheral nerve of the mouse. Injection of adriamycin into the sciatic nerve resulted in biochemical and morphologic signs of severe axonal degeneration. The biochemical evidence was based on marked increases in lysosomal enzyme activity. Acid protease and N-acetylglucosaminidase remained elevated for more than 70 days after injecting the drug. In contrast, local injection of the radiation sensitizing drug misonidazole into the peripheral nerve failed to increase lysosomal enzyme activity. PMID- 2981697 TI - Effect of insulin-induced hypoglycemia on blood-brain barrier permeability. AB - The effects of hypoglycemia on cerebrovascular permeability to the Evans blue albumin complex were studied in rats injected with 50 IU/kg, i.v. crystalline zinc insulin. One group of hypoglycemic animals was warmed to keep their body temperatures close to 37 degrees C, and the rats in the other group were allowed to become hypothermic by hypoglycemia. The arterial blood pressures of the hypoglycemic rats were continuously monitored during the coma and a significant rise in pressure was observed in most animals at the end of the coma. When glucose was administered i.v. to five animals of each group, this elevated pressure returned to normal values within 0.5 min and the animals slowly recovered normal behavior. At termination of the coma, most brains in the hypothermic hypoglycemic group showed an intensive and extensive staining by Evans blue; whereas only two brains in the normothermic hypoglycemic group showed any noticeable extravasation of Evans blue-albumin. Arterial PO2, PCO2, and pH were determined and no significant difference was found between values from animals in hypoglycemic coma and the controls. Four animals were surface-cooled and were used to examine the effects of hypothermia on blood-brain barrier permeability. These brains did not show any macroscopically evident Evans blue albumin extravasation. The results indicated that prolonged, severe hypoglycemia with hypothermia caused a profound blood-brain barrier dysfunction whereas normothermic hypoglycemia resulted in few cases of any noticeable increase in blood-brain barrier permeability. PMID- 2981698 TI - Excitability changes reveal decreased sodium permeability in neuropathy. AB - Nerve excitability was tested in isolated sciatic nerves of rats with acute uremia. Larger than normal stimulating pulses were required to elicit a maximal compound potential, whereas the relative refractory period was unaffected. The threshold range, relative to the absolute threshold, was a good measure of the decreased excitability and may be a useful clinical method. PMID- 2981699 TI - Evolution of axonal swellings in cats intoxicated with beta,beta' iminodipropionitrile (IDPN). An electrophysiological and morphological study. AB - beta,beta'-Iminodipropionitrile (IDPN) causes formation of axonal swellings in the proximal internodes of spinal motor axons. The swellings enlarge and demyelinate with the progression of the neuropathy. The correlation between axonal swellings and electrophysiologic function of motoneurons was examined in cats 2 to 35 days after initial administration of IDPN. Morphologic changes in intraspinal motor axons, occasionally observed 2 days after the first injection, became progressively more evident at later times, with enlargement at the first internode in some axons and appearance of fusiform or balloon-like axonal swellings. At 7 days axonal swellings were infrequently observed and the main structural feature was a reduction in myelin thickness in affected nerve fibers. Despite scant histopathologic changes, motoneuron action potential discharge at this time was significantly altered in latency to onset of spike and rate of rise. Abnormal motoneuron firing patterns were observed at this time. As the neuropathy progressed, both the frequency of occurrence and the size of axonal swellings increased markedly but at no time was there morphologic evidence of chromatolysis. At 14 and 35 days, (after two or five IDPN injections) action potential discharge became further altered in latency to spike onset, rate of rise, initial segment conduction time and somal-dendritic threshold. The incidence of repetitive firing increased and axonal conduction block was observed in several motoneuron recordings. The electrophysiologic changes closely resemble those reported in chromatolytic motoneurons after axotomy. The axonal swellings induced by IDPN may produce an axotomy-like condition which becomes more prominent as the neuropathy progresses but without morphologic evidence of chromatolysis. PMID- 2981700 TI - Structure of the porcine thyrotropin receptor: a 200 kilodalton glycoprotein heterocomplex. AB - We have determined that the porcine thyroidal TSH receptor is a glycoprotein heterotetramer composed of two Mr approximately 35,000 (epsilon) covalently linked subunits which interact noncovalently with two copies of delta (Mr 66,000) chains. PMID- 2981701 TI - Interaction of smooth muscle relaxant drugs with calmodulin and cyclic nucleotide phosphodiesterase. AB - Some smooth muscle relaxant drugs with an unknown mechanism of action have been tested for their interaction with calmodulin and with calmodulin-induced cyclic nucleotide phosphodiesterase (PDE) activity. The affinity of these drugs for calmodulin does not parallel their inhibitory effect on the calmodulin activation of PDE. The lack of parallelism could be due to a binding of the drugs to different sites on calmodulin; furthermore a binding of papaverine, octylonium bromide and felodipine to PDE molecule might also be considered to explain their inhibitory effect on PDE basal activity. The myolytic effect of octylonium bromide and pinaverium bromide may be due to their interaction with calmodulin dependent systems. PMID- 2981703 TI - Probing the three-dimensional structures of DNA and RNA oligonucleotides in solution by nuclear Overhauser enhancement measurements. AB - DNA and RNA oligonucleotides are ideally suited for high-resolution X-ray crystallographic and 1H-NMR studies. The solution structures of such oligonucleotides can potentially be solved using proton-proton nuclear Overhauser enhancement measurements to demonstrate the proximity of protons in space and to determine their separation, thereby enabling a comparison of the structure in the crystalline and solution states to be made. In this review we describe the general strategy for the sequential resonance assignments of oligonucleotide 1H NMR spectra, the essential prerequisite for further structural work, the approach to obtaining interproton distances from pre-steady state nuclear Overhauser enhancement measurements, and the use of interproton distances in structure determination. This is illustrated by several examples including double- and single-stranded DNA oligonucleotides as well as RNA stem and loop structures. PMID- 2981702 TI - On the mechanism of killing of Trypanosoma cruzi by human polymorphonuclear leukocytes. AB - The temperature-dependence of some processes involved in the killing of sensitized T. cruzi epimastigotes by human polymorphonuclear leukocytes (PMN) was determined. The rate of the reactions was related to the temperature of incubation according to the Arrhenius equation and the apparent energies of activation (Ea) were calculated. The Ea values separated these complex reactions into two groups: one with Ea of about 10 kcal/mol for the phagocytosis of the parasites and the release of lysosomal enzymes by PMN, and the other with Ea of about 22 kcal/mol for the cytotoxicity against sensitized T. cruzi, the rate of oxygen consumption by PMN, and the lysis of the parasites with added hydrogen peroxide. PMID- 2981704 TI - Spermine promotes the translocation of phosphatidate phosphohydrolase from the cytosol to the microsomal fraction of rat liver and it enhances the effects of oleate in this respect. AB - Spermine (0.5-2 mM) promoted the translocation of phosphatidate phosphohydrolase from the soluble to the microsomal fraction in a cell-free system derived from rat liver. By contrast, spermidine (1 mM) and putrescine (1 mM) had no significant effect on the translocation when added alone. Spermine, and to a lesser extent, spermidine, enhanced the translocating action of oleate and increased its effectiveness in transferring the phosphohydrolase from the soluble to the microsomal fraction. It is proposed that the phosphohydrolase becomes metabolically active when it combines with membranes and that polyamines might help to regulate this interaction. This could facilitate the action of fatty acids and enable cells to increase their capacity for triacylglycerol synthesis to match an increased availability of fatty acids. PMID- 2981705 TI - Monovalent nickel in hydrogenase from Chromatium vinosum. Light sensitivity and evidence for direct interaction with hydrogen. AB - Redox titrations with hydrogenase from Chromatium vinosum show that its nickel ion can exist in 3, possibly 4, different redox states: the 3+, 2+, 1+ and possibly a zero valent state. The 1+ state is unstable: oxidation to Ni(II) occurs unless H2 gas is present. The Ni(I) coordination, but not that of Ni(III), is highly light sensitive. A photoreaction occurs on illumination. It is irreversible below 77 K, but reversible at 200 K. The rate of this photodissociation reaction in 2H2O is nearly 6-times slower than in H2O, indicating the breakage of a nickel-hydrogen bond. This forms the first evidence for an H atom in the direct coordination sphere of Ni in hydrogenase and for the involvement of this metal in the reaction with hydrogen. PMID- 2981706 TI - The phosphate potential maintained by mitochondria in State 4 is proportional to the proton-motive force. AB - Evidence is presented for a proportional relationship between the extramitochondrial phosphate potential (delta Gexp) and the proton-motive force (delta mu H+) across the mitochondrial membrane in rat-liver mitochondria oxidising succinate in State 4, when delta mu H+ is varied by addition of uncouplers or malonate. This relationship was found when precautions were taken to minimise interference with the determination of delta Gexp and delta mu H+ by intramitochondrial nucleotides, adenylate kinase activity, the quenching method, and delta mu H+-dependent changes in matrix volume. A non-proportional delta Gexp/delta mu H+ relationship was obtained when these precautions were omitted. Our results do not support mosaic protonic coupling, but are not necessarily in conflict with other localised coupling schemes. PMID- 2981707 TI - Islet-activating protein discriminates between different inhibitors of thyroidal cyclic AMP system. AB - TSH-induced cyclic AMP accumulation in dog thyroid slices is inhibited by norepinephrine through an alpha 2-adrenergic receptor, by carbamylcholine through a muscarinic cholinergic receptor, and by iodide. The inhibitory effect of iodide bears on the adenylate cyclase, but the exact mechanism of its action is still unknown. It is known that norepinephrine acts through activation of the Ni subunit of the cyclase, and that carbamylcholine, activating a phosphodiesterase, acts independently of Ni. IAP (islet-activating protein) has been shown to inactivate the Ni subunit. We studied the effect of IAP on the inhibitory action of iodide, norepinephrine, and carbamylcholine on cyclic AMP accumulation in TSH stimulated thyroid slices. Incubations of 15 or 22 h, and relatively high concentrations of IAP (250 ng/ml) were necessary to demonstrate an effect of IAP on thyroid slices. We report here that, under those conditions, inhibition of cyclic AMP accumulation by norepinephrine, but not by carbamylcholine or iodide, was suppressed by IAP treatment. These results indicate that the cyclase inhibition by iodide, is either not mediated by Ni, or if mediated by Ni, involves a mode of regulation of this coupling protein that is different from that by which the other 'Ni-mediated' inhibitory hormones act on the enzyme. PMID- 2981709 TI - The Neurospora mitochondrial genome: the region coding for the polycistronic cytochrome oxidase subunit I transcript is preceded by a transfer RNA gene. AB - We have sequenced a 682 bp fragment of Neurospora crassa mitochondrial DNA to complete the sequence between the gene for cytochrome b and the unassigned reading frame, URF U. The sequence contains a gene for a cysteine tRNA. The 5' end of the 6 kb polycistronic transcript of cytochrome c oxidase subunit 1 is immediately downstream from this tRNA. This shows that also in fungal mitochondria tRNAs can be used as processing signals, whereas palindromic sequences containing double Pst I sites, also present in this region, are not used for processing. PMID- 2981708 TI - Inhibition of carcinogen-induced cellular transformation of human fibroblasts by drugs that interact with the poly(ADP-ribose) polymerase system. Initial evidence for the development of transformation resistance. AB - Two types of interactions of 13 drugs with human fibroblasts were determined: I50 of nuclear poly(ADP-ribose) polymerase, as assayed with isolated nuclei in vitro, and the non-toxic concentration of drugs that prevented carcinogen-induced cell transformation of intact fibroblasts (RCF1). In general, RCF1 was much lower than I50, and one antitransformer did not inhibit the enzyme in vitro, indicating that low-affinity enzyme inhibitory sites appear to play no role in the mechanism of prevention of cell transformation. Two enzyme inhibitors, caffeine and 1 methylnicotinamide, exhibited no antitransforming activity. Benzamide when applied in population doubling 1 induced resistance to cell transformation in population doubling 6 by carcinogens added at this stage. PMID- 2981710 TI - Persistent activity modification of phosphatidate phosphohydrolase and fatty acyl CoA synthetase on incubation of adipocytes with the tumour promoter 12-O tetradecanoylphorbol 13-acetate. AB - Incubation of rat adipocytes with the phorbol ester TPA caused a dose-dependent, persistent increase in phosphatidate phosphohydrolase activity and decrease in fatty acyl-CoA synthetase activity. Half-maximal effects were obtained with 1-2 X 10(-10) M TPA. TPA did not alter basal or noradrenaline-stimulated lipolysis. It is suggested that these two enzymes might be modified by protein kinase C. PMID- 2981711 TI - Retarded decline in poly-ADPR content and poly-ADPR synthetase activity in chicken dystrophic muscle. AB - The activity of poly-ADPR synthetase declines just after hatching in normal chick muscle nuclei. However, in dystrophic chick muscle nuclei it decreases 5 weeks after hatching. A delayed decrease in the amount of poly-ADPR is also observed in dystrophic chick muscle nuclei. These observations suggest that dystrophic muscle follows an abnormal developmental program. PMID- 2981712 TI - The presence and orientation of ecto-5'-nucleotidase in rat liver lysosomes. AB - Purified rat liver lysosomes contained 5'-nucleotidase activity which was 92 +/- 2% [4]latent. This latency was lost in response to a permeant sugar at a similar rate to that of the lysosomal marker enzyme beta-N-acetylglucosaminidase indicating that the 5'-nucleotidase was genuinely located in the lysosome and not a plasma membrane contaminant. Lysosomal 5'-nucleotidase exhibited the following properties characteristic of ecto-5'-nucleotidase inhibition by specific polyclonal antibodies: binding to a monoclonal antibody; inhibition by 1 mmol/1 alpha beta-methylene ADP; immunoreactive subunits of 70 and 38 kDa. Lysosomes in addition contained immunoreactive species of intermediate molecular mass. PMID- 2981713 TI - Effects of prostaglandin E2 and cholera toxin on apical sodium uptake in thyroid epithelial cells: role of cAMP. AB - When cultured in tissue culture polystyrene dishes, porcine thyroid cells formed polarized monolayers. Their apical pole was oriented towards the culture medium. Influx of sodium 22 (5 min) through the apical surface was partially inhibited by amiloride. Amiloride-sensitive Na uptake was increased about 3-fold by prostaglandin E2 (1 microM, 30 min) and by cholera toxin (0.01 micrograms/ml, 1 h). This increase, which was also obtained with forskolin and 8-(4 chlorophenylthio)adenosine-3':5'-monophosphate, cyclic (8-chloro-cAMP), is probably a consequence of the increase of the intracellular cAMP level by prostaglandin and cholera toxin. PMID- 2981714 TI - Subcellular localization and some properties of the enzymes hydrolysing inositol polyphosphates in rat liver. AB - The hydrolysis of inositol [32P]trisphosphate (IP3) and inositol [32P]bisphosphate (IP2) has been examined in subcellular fractions of rat liver. IP3 was degraded by an enzyme located in the plasma membrane which did not degrade IP2. Cytosolic fractions were found to degrade both IP2 and IP3. The IP3 phosphatase activity of liver plasma membranes displayed a neutral pH optimum, was Mg2+ dependent and was not inhibited by high concentrations of Li+. Half maximal activity of the enzymes hydrolysing IP3 and IP2 was obtained with substrate concentrations in the range 1-2 microM. The significance of these observations to the proposed Ca2+-mobilizing role of IP3 is discussed. PMID- 2981715 TI - Tumour-promoting phorbol esters inhibit agonist-induced phosphatidate formation and Ca2+ flux in human platelets. AB - Tumour-promoting phorbol esters (phorbol-12-myristate-13-acetate, PMA; phorbol 12,13-dibutyrate, PDBu) but not 4 beta-phorbol, activate protein kinase C. Using human platelets pre-labelled with quin2 or 32PO4 we examined the effects of these compounds on human platelet cytosolic free Ca2+ ([Ca2+]i) and on [32P]phosphatidic acid ([32P]PtdOH). PMA and PDBu, but not 4 beta-phorbol inhibited thrombin-, PAF- and vasopressin-induced elevation of [Ca2+]i and [32P]PtdOH formation. It is suggested that protein kinase C may act to terminate the transduction processes that link receptor occupancy to cellular activation. PMID- 2981716 TI - Conservation from rat to human of cytosolic phosphoenolpyruvate carboxykinase and the control of its gene expression. AB - Structural conservation of cytosolic phosphoenolpyruvate carboxykinase protein and mRNA sequence was found in all species examined from rodents to human. The mitochondrial isoenzyme, in all species tested, represents a distinct protein. Moreover, irrespective of the ratio of cytosolic to mitochondrial isoenzyme, cytosolic phosphoenolpyruvate carboxykinase activity in the human as in the rat is controlled at the level of gene expression and through the same multiple hormonal stimulation. This evolutionary conservation of the cytosolic phosphoenolpyruvate carboxykinase structure and mode of regulation supports the enzymes' physiological importance in mammals. PMID- 2981717 TI - Activation of protein kinase C in neutrophil cytoplasts. Localization of protein substrates and possible relationship with stimulus-response coupling. AB - Treatment of enucleated, granule-free neutrophil cytoplasts with the protein kinase C activator phorbol 12-O-myristate-13-acetate (PMA) causes an increased 32P-incorporation into a variety of polypeptides. Permeabilization of PMA stimulated, 32P-labeled cytoplasts by 0.01% digitonin fully releases the majority of these phosphorylated proteins. A statistically significant correlation is found between the extent of PMA-induced activation of generation of superoxide anion (O2-) and the phosphorylation of a cytosolic polypeptide with an apparent Mr of 46,000, whose 32P-labeling is also enhanced by the treatment of cytoplasts with 1-oleyl-2-acetylglycerol, the Ca2+ ionophore ionomycin or latex beads. Furthermore, treatment of cytoplasts with the protein kinase C inhibitor trifluoperazine markedly inhibits the 32P-labeling of proteins in the 40 000 Mr range, including the 46 kDa polypeptide, and almost totally abolishes the activation of O2- production by PMA. PMID- 2981718 TI - Vasoactive intestinal peptide receptor activity in human fetal enterocytes. AB - Functional and specific receptors for vasoactive intestinal peptide (VIP) (determined by their capacity to bind 125I-VIP and activate adenylate cyclase) and cyclic AMP-dependent phosphodiesterase activities were characterized in enterocytes of human fetal small intestine between 18 and 23 weeks of gestation. Half-maximal stimulation of the cyclase and inhibition of 125I-VIP binding in membrane preparations were respectively observed at 1.4 and 5 X 10(-10) M VIP. The peptides structurally related to VIP activated the cyclic AMP generating system at pharmacological doses (10(-7) M and above) in the following order of potency: VIP greater than PHI greater than GRF greater than secretin. Other peptides or test substances, including GIP, pancreatic glucagon, somatostatin-14, gastrin, CCK, neurotensin, pancreatic polypeptide, PYY, substance P, histamine and isoproterenol are inactive in this system, while the ubiquitous adenylate cyclase activators NaF, forskolin and prostaglandins were effective. These results, combined with the appearance of intestinal VIP in nerve fibers at 8 weeks and with the morphological and enzymatic maturation at 9-12 weeks of the intestinal mucosa, indicate that this neuropeptide may regulate either the differentiation or function of enterocytes during the early development of human intestinal mucosa. PMID- 2981719 TI - Cyclic AMP stimulates dephosphorylation of specific proteins in intact S49 mouse lymphoma cells. AB - Two-dimensional gel electrophoresis of proteins labeled with 32Pi in S49 mouse lymphoma cells revealed five phosphoproteins that were rapidly and reversibly dephosphorylated in response to elevation of cyclic AMP (cAMP). Under basal conditions, labeling of at least two of these proteins was limited by slow turnover of protein-bound phosphate. The rapid cAMP-mediated dephosphorylation of these species was attributable, therefore, to stimulation of a specific phosphoprotein phosphatase. PMID- 2981720 TI - Polyadenylation of histone mRNA in Xenopus oocytes and embryos. AB - Oogenesis of amphibians is an atypical situation in which histone mRNA is polyadenylated. The poly(A) tract on H4 mRNA has been examined by S1 nuclease analysis. Throughout oogenesis the poly(A) tract is very short, and nonexistent on some mRNA molecules. The poly(A) tract is completely removed during maturation of the oocyte, and is absent in embryos and cultured cells. PMID- 2981721 TI - Reduction of the metallochromic indicators arsenazo III and antipyrylazo III to their free radical metabolites by cytoplasmic enzymes. AB - At a concentration much lower than that usually employed for measuring cytosolic ionized Ca2+ concentrations, arsenazo III underwent a one-electron reduction by rat liver cytosolic fraction or a hypoxanthine-xanthine oxidase system to produce an azo anion radical metabolite. NADH, NADPH, N1-methylnicotinamide, hypoxanthine, and xanthine, in that order, could serve as a source of reducing equivalents for the production of this free radical by the cytosolic fraction. The steady-state concentration of the azo anion radical and the arsenazo III stimulated O2 consumption were enhanced by calcium and magnesium. Antipyrylazo III was ineffective in increasing O2 consumption by rat liver cytosolic fraction and gave a much weaker ESR signal of an azo anion radical with both the liver cytosolic fraction, in the presence of NADH, and the hypoxanthine-xanthine oxidase system. PMID- 2981722 TI - Identification of the peptide bond cleaved during activation of human C1r. AB - CNBr cleavage of unreduced proenzyme C1r yielded fragment CP2b, isolated by gel filtration and high-pressure gel permeation chromatography. This fragment (approximately Mr 55 000) comprised at least 4 disulphide-linked peptides, which were separated by gel filtration after reduction and alkylation. Peptide CP2bRA4, overlapping the A- and B-chain regions in proenzyme C1r was digested by V8 staphylococcal protease, and the digest separated by reversed-phase HPLC. N terminal sequence analysis of peptide CP2bRA4SP9 established that C1r activation involves the cleavage of a single Arg-Ile bond, located in the sequence: ... Gln Arg-Gln-Arg-Ile-Ile-Gly-Gly ... . PMID- 2981723 TI - Proteolysis of mitochondria in reticulocytes during maturation is ubiquitin dependent and is accompanied by a high rate of ATP hydrolysis. AB - The ATP-dependent breakdown of mitochondria-containing stroma proceeds via the ubiquitin-requiring pathway. The proteolysis is linked to a large ATP-cleaved consumption amounting to 1 ATP per peptide bond or more. Proteins of mitochondria containing stroma are much better substrates of ATP-ubiquitin-dependent proteolysis than heat-denatured ones. Hemin suppresses both proteolysis and ATP hydrolysis. PMID- 2981724 TI - Phosphorylation of hormone-sensitive lipase by cyclic GMP-dependent protein kinase. AB - In intact rat adipocytes hormone-sensitive lipase has been shown to be phosphorylated on serine residues in two different phosphorylation sites: a regulatory site phosphorylated by cyclic AMP-dependent protein kinase and a basal site, which does not directly affect the enzyme activity, phosphorylated by cyclic AMP-independent protein kinase(s) [(1984) Proc. Natl. Acad. Sci. USA 81, 3317-3321]. Cyclic GMP-dependent protein kinase catalyzed the phosphorylation of the same two phosphorylation sites on the isolated enzyme, at serine residues. Both sites were phosphorylated at about the same rate, with the hormone-sensitive lipase activity concomitantly enhanced. PMID- 2981725 TI - Relationship of phosphorylation to the oligomerization of SV40 T antigen and its association with p53. AB - The potential significance of the phosphorylation of SV40 large T antigen for oligomers and complexes with the cellular protein p53 was investigated. We observed that T antigen oligomers remain stable after enzymatic dephosphorylation by alkaline phosphatase up to 80%. Separate analysis of free and p53-bound T antigen revealed a considerably lower phosphorylation of the p53-bound subclass. Therefore, a simple correlation between the overall phosphorylation of T antigen and the formation of oligomers and T-p53 complexes is highly unlikely. PMID- 2981726 TI - Involvement of lysine residues in the binding of hGH and bGH to somatotropic receptors. AB - The biological activities of human (hGH) and bovine (bGH) growth hormone derivatives obtained by chemical modification of the lysine residues were studied by radioreceptor assays using rabbit liver homogenates for somatotropic activity (SA). Control treatment with BH4 had a very slight effect on the SA, whereas the methylation and ethylation drastically reduced the activity of both hormones. Guanidination of these hormones and even acetimidination at a lower rate are accompanied by a considerable loss of biological activity. These results show the involvement of lysine residues in the interaction of hGH and bGH with somatotropic receptors. The structure-function relationship of these molecules is discussed, suggesting that the lysine or arginine residues in positions 41, 64, 70 and 115 might be particularly implicated. PMID- 2981727 TI - Antiviral activity of phorbol myristate acetate and possible relationships with interferon action. AB - Signal-induced turnover of membrane phospholipids represents a fundamental transducing mechanism that induces a signal cascade resulting in mobilization of calcium, activation of protein kinase C by diacylglycerol, release of arachidonic acid and stimulation of cyclic GMP production. In this pathway tumor-promoting phorbol esters such as phorbol myristate acetate (PMA) may substitute for diacylglycerol. The interferon-like antiviral effect of PMA described here suggests that the inositol phospholipid-diacylglycerol-protein kinase C signal transducing mechanism may be involved in interferon action. PMID- 2981728 TI - Anatomy of the corticotropin-releasing factor and opiomelanocortin systems of the brain. AB - Using antiserums generated against the neuropeptides corticotropin-releasing factor (CRF), beta-endorphin, and adrenocorticotropin, we have immunocytochemically mapped the organization of these hormones throughout the brain. The distribution of CRF perikarya is widespread throughout the forebrain, brain stem, and spinal cord. The central opiomelanocortin system is represented by two separate pools of neuronal perikarya: the hypothalamic arcuate pool, and the medullary (nucleus tractus solitarius) pool. From these groups fibers distribute widely throughout forebrain and brain stem. A conspicuous codistribution pattern exists between the CRF perikarya and opiomelanocortin fibers. Analysis of these systems within the brain stem and spinal cord reveals innervation and distribution patterns within regions containing catecholamine systems, areas associated with regulation of sympathetic and parasympathetic outflow, cardiovascular and respiratory homeostatic centers, and regions associated with stress, autonomic homeostasis, and pain modulation. By virtue of these extensive brain-stem distributions, it is suggested that an anatomical substrate exists for participation of these peptide systems in the integration of certain autonomic, visceral, and homeostatic mechanisms. PMID- 2981729 TI - Arachidonic acid metabolites and endothelial injury: studies with cultures of human endothelial cells. AB - Human endothelial cells in culture can metabolize arachidonic acid to prostaglandin (PG) I2 (prostacyclin), PGE2, and PGF2 alpha, and to various nonpolar products. Similar metabolites are formed by cultured pulmonary arterial and venous cells and umbilical venous cells. In studies with umbilical venous endothelial cells, PG formation was stimulated by mechanical agitation or by treatment with histamine or melittin. Histamine stimulated prostacyclin release into cell culture media without damaging the cells, but melittin injured the cell membrane as indicated by the loss of 51Cr from labeled cells. Media collected from mechanically agitated cells inhibited the release of chromium by melittin. Heating to 56 C or acidification of the media did not reverse this protective effect. Prostacyclin added to labeled cells before challenge with melittin partially inhibited 51Cr release, but PGE2 or leukotrienes B4, C4, and D4 did not. In experiments with pulmonary endothelial cells, histamine augmented the release of hydroxyeicosatetraenoic acids (HETEs) and other nonpolar metabolites. The major nonpolar product comigrated on high-pressure liquid chromatography with di-HETEs. 11-HETE and 15-HETE were identified in media from histamine-stimulated cells by comigration with standards. Neither of these compounds nor 12-HETE protected endothelial cells from melittin-induced injury. It is not yet known if any arachidonic acid metabolites can modulate endothelial cell injury. PMID- 2981730 TI - Leukotrienes: their formation and role as inflammatory mediators. AB - The leukotrienes (LTs) are a novel group of biologically active mediators derived from arachidonic acid via lipoxygenase enzymes. LTB4 is a potent chemotactic agent for polymorphonuclear leukocytes and in vivo may mediate inflammatory reactions by inducing leukocyte recruitment by mediating indirectly vascular permeability charges and by modulating pain responses. LTC4 and LTD4 collectively account for the biological activity known as slow-reacting substance of anaphylaxis and are potent smooth muscle contracting agents. They may mediate inflammatory reactions by producing changes in blood flow and increases in vascular permeability. Evidence for LT involvement in a number of pathological conditions including diseases such as asthma, psoriasis, ulcerative colitis, and gout is now accumulating. PMID- 2981731 TI - Effect of neutrophil migration induced by leukotriene B4 on protein permeability in sheep lung. AB - Although several types of acute lung microvascular injury require active participation of circulating leukocytes (presumably neutrophils), the relationship between neutrophils and microvascular injury and edema is controversial. We attempted to answer the question, Does neutrophil migration in response to a chemical signal alter endothelial permeability? If so, is this to be regarded as a significant control mechanism for microvascular protein permeability? Because of our interest in pulmonary injury, we attempted to induce neutrophil chemotaxis into the lungs of anesthetized sheep by using leukotriene B4 (LTB4) (2 nmol/ml, 50 ml total) placed in the alveoli of one caudal lung lobe through a fiberoptic bronchoscope. Measurements of pulmonary hemodynamics, lymph dynamics, and alveolar liquid dynamics were made. Compared to chemotaxis into skin, neutrophil migration into alveoli was slow. LTB4 caused a small increase in lung lymph and protein flow, which was slightly more than that in saline controls. Likewise, excess plasma protein entering the alveolar liquid was slightly increased over saline controls. Both results indicate a small increase in liquid and protein flow across the endothelial and alveolar epithelial barriers in response either to the LTB4 or to the neutrophil chemotaxis. Using a worst case analysis, the slight increase in liquid and protein flow across the endothelium and epithelium may be related to quantal leakage of plasma during neutrophil transit, but there is no evidence for any persistent change in microvascular or epithelial permeability. PMID- 2981732 TI - Pulmonary microvascular effects of arachidonic acid metabolites and their role in lung vascular injury. AB - Arachidonic acid metabolites produced by cyclooxygenase and lipoxygenase pathways affect pulmonary transvascular fluid and protein fluxes after pulmonary microvascular injury. Some of these products may contribute to the increase microvascular endothelial permeability whereas others may increase pulmonary microvascular filtration pressure. Prostaglandin (PG) E2, PGF2 alpha and cyclic endoperoxides increase microvascular pressure and thus increase the transvascular fluid filtration rate. Thromboxanes increase microvascular pressure and in addition may promote neutrophil adherence to endothelium and platelet aggregation, whereas prostacyclin has opposing actions. The cysteine-containing leukotrienes (LTs) (LTC4, LTD4, and LTE4) increase pulmonary microvascular pressure via a thromboxane-mediated mechanism, and LTB4 may increase pulmonary vascular permeability. Arachidonic acid metabolites do not appear to alter directly pulmonary endothelial membrane permeability but may contribute to the increased permeability by their actions on blood-formed elements. The pulmonary vasoconstrictor arachidonic aid metabolites increase microvascular hydrostatic pressure and may thereby enhance the degree of pulmonary edema. PMID- 2981733 TI - Current concepts of volume receptor regulation of vasopressin release. AB - For 2 decades evidence accumulated that supported the Gauer-Henry hypothesis tying blood volume changes to the control of vasopressin (VP). By the mid 1970's this left atrial hypothesis was generally accepted even though there was a significant body of conflicting data and several definitive questions that had not been answered. In the last decade numerous investigations have addressed these questions and although there are better answers for some, others remain elusive. Pertinent work during the past 10 years is reviewed in the framework of some of these questions. The emphasis is placed on the location of the primary volume receptors, determination of the threshold for effects on VP, the degree of volume-pressure receptor and volume-osmotic receptor interactions, and species differences. PMID- 2981734 TI - The use of antagonists of vasopressin in studies of its physiological functions. AB - Antagonists of the vasopressor response to vasopressin (VP) have proved useful for probing the contributions of endogenous VP to cardiovascular regulation. Studies with such antagonists show that VP can exert substantial cardiovascular effects even when circulating levels are relatively low, as they are after moderate dehydration. Antagonists that also block antidiuretic responses to VP have recently become available. These promise to be equally useful for assessing the importance of VP in limiting water excretion under a variety of circumstances. Although many of the antagonists of the vasopressor response are relatively specific, the early antidiuretic antagonists were not. They block both vasopressor and antidiuretic receptors. Recently we have made progress in designing analogs that are more specific antagonists of the antidiuretic response. This suggests that it should be possible to produce specific antagonists of the antidiuretic response that would provide more incisive pharmacological probes and could become useful therapeutic agents. PMID- 2981735 TI - Enkephalins and endorphins as modifiers of the immune system: present and future. AB - This paper summarizes the data related to the relations between enkephalins and endorphins and the immune system. It is shown that lymphocytes have surface receptors for endorphins and enkephalins. Furthermore, endorphins and enkephalins can influence several immune functions such as antibody synthesis, lymphocyte proliferation, and natural killer cytotoxicity. It is thus possible that the receptors play a functional role. In view of these observations, several hypotheses are proposed. Endorphins and enkephalins are physiological regulators of the immune response (two-hit opioid peptide lymphocyte receptor hypothesis) and they are humoral mediators between the central nervous system and the immune system. They may play a pathogenic role in a variety of diseases with primary or secondary immunological defects. Finally, enkephalins and endorphins can be considered as immunomodulators and modifiers of the biological response and as such may become a tool in the field of immunotherapy. PMID- 2981736 TI - Mechanisms by which stressful stimuli activate the pituitary-adrenal system. AB - Stress-induced activation of the pituitary-adrenal system appears to be mediated predominantly by hypothalamic neurons containing ovine corticotropin-releasing factor (oCRF) and vasopressin in the region of the paraventricular nucleus (PVN). Other mediators such as catecholamines, serotonin, angiotensin, and "tissue CRF" may also play a role under certain conditions. Several weeks after ablation of the PVN the response to stress is largely restored while the plasma adrenocorticotropin rise normally elicited by adrenalectomy is still strongly inhibited and immunoreactive oCRF is depleted from the stalk-median eminence region. PMID- 2981737 TI - Role of epinephrine and vasopressin in the control of the pituitary-adrenal response to stress. AB - In addition to corticotropin-releasing factor (CRF) and structurally related peptides, arginine vasopressin (AVP), oxytocin, angiotensin II, vasoactive intestinal polypeptide, peptide histidine isoleucinamide, epinephrine (E), and norepinephrine induce secretion of adrenocorticotropin (ACTH) from corticotropic cells in vitro. The apparent affinity and intrinsic ACTH-releasing activity of these substances are lower than those of CRF. These substances can also act synergistically with CRF. In this paper the role of catecholamines and AVP in the control of ACTH release is discussed. Infusion i.v. of E increases plasma ACTH and corticosterone to levels that are normally found during stress. E-induced stimulation of pituitary-adrenal activity is mediated by beta adrenoceptors and involves release of CRF, because it can be prevented by beta-adrenoceptor blockers and by destruction of CRF neurons (hypothalamic lesions), blockade of CRF release (chlorpromazine, morphine, and Nembutal), or administration of CRF antiserum. Although stress can cause a vast increase in plasma E, circulating E is not essential for the acute stress-induced release of ACTH because blockade of beta (or alpha) adrenoceptors, administration of chlorisondamine, or extirpation of the adrenal medulla and sympathectomy do not prevent the pituitary-adrenal response to stress. In contrast, circulating E plays a major role in the release of intermediate-lobe peptides during emotional stress. Studies of the role of AVP in pituitary-adrenal control by the use of pressor receptor (V1) antagonists are not valuable because of the ineffectiveness of such antagonists in blocking AVP induced release of ACTH from corticotropic cells in vitro. Treatment of rats with an antiserum to AVP reduces the ACTH response to stress. We conclude that AVP has an important role in stress-induced activation of the pituitary-adrenal system, possibly by potentiating the effects of CRF. PMID- 2981738 TI - Neural interaction in control of adrenocorticotropin. AB - Multiple afferent pathways underlie the control of adrenocorticotropin (ACTH). The variety of sensory stimuli that elicit ACTH release indicate that neural interaction and convergence within the central nervous system contribute to this control system. The site and nature of neural interactions contributing to ACTH release are unknown. However, recent evidence suggests that some of the sensory convergence and temporal plasticity within the system that controls ACTH release may exist at the brain-stem level. Within the hypothalamus, different areas may preferentially activate the release of different releasing factors so that additional convergence and interaction may occur within the median eminence or the pituitary. PMID- 2981739 TI - Closed-loop feedback control of the nyctohemeral rise in adrenocortical system function. AB - We tested the hypothesis that the increase in adrenocortical system activity is controlled by an error signal generated by the difference between a central reference, or set point, value and some aspect of circulating corticosterone levels in rats maintained on a 12-h light, 12-h dark cycle. The test rats were treated with cyanoketone, an inhibitor that blocks conversion of pregnenolone to progesterone; control rats were treated with vehicle. Test and control rats were fitted with arterial cannulas and, after recovery, continuously collected blood was sampled for plasma corticosterone at 5-min intervals over 2-h periods at five times during the day. Samples were collected at the end of each period for plasma adrenocorticotropin (ACTH). Corticosterone and ACTH levels were similar in the two groups of rats at the nadir (0-2 h after lights-on) and at the peak (0-2 h after lights-off) of the diurnal rhythm. During the time of the maximal diurnal rise in corticosterone (8-10 and 10-12 h after lights-on), test rats had lower mean corticosterone and higher ACTH levels than controls. Infusion of 8 micrograms corticosterone during the 2 h before lights-out did not significantly elevate mean corticosterone levels but decreased end ACTH levels in test rats, whereas similar infusions in control rats resulted in decreased mean corticosterone levels and no change in end ACTH. We conclude that the nyctohemeral rise in ACTH is driven by an error signal resulting from the difference between a central set point value and some aspect of plasma corticosterone levels. PMID- 2981740 TI - Effects of corticotropin-releasing factor, neurohypophyseal peptides, and catecholamines on pituitary function. AB - In freely moving rats, ovine corticotropin-releasing factor (CRF) and rat CRF, which are equipotent in stimulating adrenocorticotropin (ACTH) release, can exert this effect after either i.v. or intracerebroventricular (i.c.v.) administration. Oxytocin and epinephrine also elevate plasma ACTH levels, an effect that is abolished by immunoneutralization of endogenous CRF. Inasmuch as oxytocin and epinephrine show additivity with CRF, these results suggest that these two secretagogues stimulate ACTH secretion in vivo by interacting with endogenous CRF. Apart from its effect on ACTH release, CRF injected i.c.v. markedly inhibits luteinizing hormone (LH), but not follicle-stimulating hormone, secretion in rats in the absence of circulating levels of steroids. A similar effect is observed after i.c.v. administration of sauvagine, a peptide analogous to CRF, whereas arginine vasopressin exhibits lower potency and shorter duration of action than CRF. Because these peptides do not modify LH release by cultured pituitary cells, they probably lower plasma LH levels through centrally mediated mechanisms. These results indicate that CRF can exert a broad spectrum of action to regulate pituitary function directly or indirectly. PMID- 2981741 TI - Clinical studies with synthetic ovine corticotropin-releasing factor. AB - Ovine corticotropin-releasing factor (oCRF) stimulates increased plasma immunoreactive adrenocorticotropin (IR-ACTH) and IR-cortisol at threshold, half maximal, and maximal doses of 0.01-0.03, 0.3-1, and 3-10 micrograms/kg, respectively. Side effects occur with increasing frequency, severity, and duration at doses above 1 microgram/kg. oCRF has a prolonged duration of action, at least in part because of the long circulating half-life of intact oCRF in plasma. Increasing doses of oCRF given in late afternoon progressively diminish the next morning's circadian rise in plasma IR-ACTH in normal subjects, but not in Addisonian patients or subjects receiving metyrapone, indicating that prolonged oCRF-induced hypercortisolemia is the cause. Plasma IR-lipotropins and IR-beta-endorphin rise and fall concomitantly with IR-ACTH after oCRF injection. Arginine vasopressin increases the IR-ACTH response to oCRF fourfold when given simultaneously with oCRF. Cushing's disease patients respond variably, suggesting that oCRF may not be a very useful diagnostic agent in Cushing's syndrome. However, the combination of oCRF with growth hormone-releasing factor, gonadotropin-releasing hormone, and thyrotropin-releasing hormone appears to provide a rapid and useful test of combined anterior pituitary function. PMID- 2981742 TI - Measurement of hypothalamic corticotropin-releasing factors in hypophyseal portal blood. AB - Among the putative hypothalamic hormones that can influence the release of adrenocorticotropin (ACTH) in vitro, the 41-amino-acid corticotropin-releasing factor, oxytocin, vasopressin, and epinephrine were identified in hypophyseal portal plasma. Measurement of these substances in several situations associated with changes in ACTH secretion provided data consistent with a physiological role for all four hormones and suggested that control of the hypothalamo-hypophyseal adrenal axis can be accomplished in different ways in response to different types of stimuli. PMID- 2981743 TI - Localization, colocalization, and plasticity of corticotropin-releasing factor immunoreactivity in rat brain. AB - The generation of antiserums against a peptide that has met the criteria predicted for corticotropin-releasing factor (CRF) has allowed the immunohistochemical localization of CRF immunoreactive neurons in the rat brain. Although CRF-stained cells have been found to be widely distributed in the central nervous system, attention has focused on neurons in the paraventricular nucleus of the hypothalamus (PVH), which is now acknowledged to be the principal source for delivery of CRF to the hypophyseal portal system. Some 2000 CRF stained neurons can be counted in the PVH of the colchicine-treated rat, and there is evidence that enkephalin, PHI, and neurotensin coexist with CRF in subsets of parvocellular neurons. Consistent with the established negative feedback effects of adrenal steroids on CRF production and release, adrenalectomy enhances CRF immunoreactivity in parvocellular neurosecretory neurons in the PVH. In addition, immunoreactive vasopressin can be demonstrated in a majority of CRF stained parvocellular neurons after adrenalectomy, which suggests a form of plasticity that allows for synergy of the two peptides in stimulating adrenocorticotropin secretion. The effects of adrenalectomy appear to be glucocorticoid-dependent, and specific to these peptides and this cell type. A survey of neural inputs to the hypophyseotropic zone of the PVH suggests potential substrates for the control of CRF release and/or synthesis by interoceptive stimuli, by the limbic region, and by a number of cell groups in the basal forebrain. Finally, CRF may also participate in other (nonadenohypophyseal) modes of regulation that are represented in the PVH. Thus, CRF immunoreactivity has been demonstrated in a discrete subset of oxytocinergic magnocellular neurosecretory neurons that project to the posterior pituitary, and in a small fraction of cells in the parvocellular division that project to cell groups in the brain stem and spinal cord that are associated with the control of autonomic functions. PMID- 2981744 TI - Corticotropin-releasing factor: effects on the autonomic nervous system and visceral systems. AB - Corticotropin-releasing factor (CRF) acts within the central nervous system to modify the activity of the sympathetic and parasympathetic nervous systems, cardiovascular function, endocrine-pancreatic function, and carbohydrate metabolism. These changes in animal physiology induced by CRF provide a tenable basis for hypothesizing that this peptide may be physiologically involved in integrating the neuroendocrine, autonomic, cardiovascular, and metabolic responses to stressful stimuli. This hypothesis is supported by studies demonstrating that glucocorticoids or CRF receptor antagonists alter both the neuroendocrine and autonomic responses to stress. PMID- 2981745 TI - Eicosanoids as mediators of ischemia and shock. AB - A variety of eicosanoids are produced in ischemic and circulatory shock. Many of these constrict arteries, induce platelet aggregation or adherence of other blood cells to the vasculature, and contribute to increased membrane permeability. Thromboxane A2, leukotriene C4, and leukotriene D4 fulfill all the criteria stipulated for humoral mediators of ischemia and shock. Moreover, pharmacologic modulation of these mediators by either specific inhibition of their synthesis or antagonism of their arteries at their receptor sites protects against tissue and cell damage during ischemia and shock as well as enhances survival in these life threatening states. PMID- 2981746 TI - Regulation of branched chain alpha-ketoacid dehydrogenase complex by phosphorylation-dephosphorylation. Minisymposium report. PMID- 2981748 TI - Steroidal regulation of endometriosis tissue: lack of induction of 17 beta hydroxysteroid dehydrogenase activity by progesterone, medroxyprogesterone acetate, or danazol. AB - Cytosol and nuclear estrogen receptors were detected in 73% and 89% of untreated endometriosis specimens, respectively. The corresponding figures for cytosol and nuclear progestin receptors were 94% and 100%, respectively. Compared with the endometrium, the concentrations in endometriosis tissue were low. The anatomic site, the severity of the disease, and the phase of the menstrual cycle had no significant influence on receptor concentrations in endometriosis tissue. The activities of 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) did not increase during the luteal phase. Danazol for 1, 3, 7 to 11, and 18 to 30 days, or medroxyprogesterone acetate for 5 days, had no effect on receptor concentrations or 17 beta-HSD activity in endometriosis tissue. The frequent presence of the receptors suggests that endometriosis lesions are under the control of steroid hormones. The lack of effect by progesterone and progestins on 17 beta-HSD shows that this regulation is different in endometriosis tissue and in the endometrium. PMID- 2981747 TI - The human adrenergic neurovascular mechanism. AB - The major features of adrenergic neurotransmission established in animals have been observed in one blood vessel or another from the human. However, the detailed mechanism in one particular human blood vessel or vascular sample from a particular regional bed has not yet been carefully compared with a corresponding vessel from nonhuman species. There is probably more known about cerebral blood vessels than others, and if our knowledge of this bed is at all representative of what occurs elsewhere, then any projections regarding the human from animal studies are potentially very misleading. The little evidence that is available suggests that generalization regarding human vessels is going to be difficult. Yet this very difficulty emphasizes the need for study, not only from the normal and healthy subject, but from the diseased. The human circulation represents a source of vascular tissue whose analysis will reveal much more about the circulation and its change with circumstances, including disease, of interest to the scientist and the physician than can ever be achieved by experimentation with animal tissues. PMID- 2981749 TI - Insulin gene analysis in a family with maturity-onset diabetes of the young. AB - The insulin gene locus has been studied in a large kindred with maturity-onset diabetes of the young (MODY) characterized by hypoinsulinemia. DNA was isolated from peripheral leukocytes of 42 family members and 5 spouses. A highly polymorphic region in the 5'-flanking portion of the human insulin gene provided an opportunity for linkage analysis. The presence of three different length polymorphisms of + 1600 base pairs (bp), - 50 bp, and - 150 bp different from the common size allele allowed haplotype assignment of insulin alleles. The hypothesis of linkage was tested by calculating the log of the ratio of the likelihood of the hypothesis of linkage to that of the hypothesis of nonlinkage (LOD score) at a given recombination distance between the insulin polymorphism and the diabetes locus. At a recombination frequency of 0.0, the LOD score was - 14.50 and, therefore, the hypothesis of tight linkage can be strongly rejected. This report is the third study of the relationship between the insulin locus and MODY; however, it is the first report in which a formal linkage analysis indicates with a high degree of probability no linkage between the insulin locus and hypoinsulinemia in a family. Because MODY is a heterogeneous disorder, it may be that different genotypes result in a composite phenotype. The lack of linkage between an insulin allele and MODY in a total of four families studied, however, suggests that the insulin locus is probably not a marker for the MODY phenotype. These results do not exclude the possibility that the insulin locus may be involved in the etiology of other forms of NIDDM. PMID- 2981750 TI - Cytosolic insulin-degrading activity in islet-derived tumor cell lines and in normal rat islets. AB - RIN-m cells, cultured from a rat insulinoma, not only bind and secrete but also degrade insulin (Diabetes 1982; 31:521-31). The insulin-degrading activity resides in the cytosol and is similar to the insulin-specific proteases previously described in muscle and other tissues. It has an apparent Km of 0.15 microM for porcine insulin in crude cell-free extracts, a competitive inhibition constant for proinsulin that is close to the Km, and a lower but measurable affinity for glucagon. The enzyme is inactive at pHs below 6.0, indicating that it is not lysosomal, is completely inhibited by N-ethylmaleimide, and exhibits apparent competitive inhibition constants (microM) for the following peptides: desoctapeptide insulin, 0.043; guinea pig insulin, 0.048; proinsulin, 0.64; insulin B-chain, 1.17; glucagon, 7.0; and cyclic somatostatin, 8.6. Highly active insulin-degrading activity was found using cell suspensions of 22 cloned and 8 subcloned cell lines derived from RIN-m as well as 11 other continuous cell lines derived from a variety of nonislet tissues of rat, mouse, and human origin. Homogenates of the original rat islet tumor and cytosol of normal rat islets also contained insulin-degrading activity. Although insulin protease is present in a variety of tissues, it may have an additional regulatory function in cells that are actively synthesizing, storing, and secreting insulin. PMID- 2981751 TI - Administration of silica prevents diabetes in BB-rats. AB - Administration of silica to young BB-rats almost completely prevented the development of spontaneous diabetes. Only 1 of 31 silica-treated rats developed hyperglycemia, whereas 9 of 31 in the untreated group did so. Since silica is highly specific in its action against macrophages, our observations indicate an important role of these cells in the pathogenesis of the disease. PMID- 2981752 TI - Peripheral neuropathy in Crohn's disease patients treated with metronidazole. AB - Thirteen pediatric patients with Crohn's disease, aged 12-22 yr, were studied to assess the prevalence of peripheral neuropathy due to oral metronidazole. After 4 11 mo of therapy, 11 of 13 patients (85%) had a sensory peripheral neuropathy, determined by abnormal neurologic examinations or reduced nerve conduction velocities, or both. Only 6 of the 11 patients were symptomatic. Nine of 11 patients with peripheral neuropathy had their metronidazole discontinued and 2 had the dose reduced to less than 10 mg/kg X day. Follow-up evaluations of the 9 patients whose metronidazole had been discontinued 5.5-13 mo earlier demonstrated complete resolution of the peripheral neuropathy in 5, improvement in 3, and no change in 1. In the 2 patients whose metronidazole dose was reduced, 1 showed worsening and 1 showed complete resolution of the neuropathy after 10-12 mo of continued therapy. PMID- 2981753 TI - Serum levels of estrogens and testosterone in cirrhotic men with and without hepatocellular carcinoma. AB - Serum levels of estrogens and testosterone were measured in 25 male patients with hepatocellular carcinoma and associated cirrhosis of the liver and in another 25 male patients with cirrhosis only. The two groups were statistically comparable in terms of age distribution, duration of liver disease, incidence of alcohol abuse, incidence of hepatitis B surface antigenemia, and grade of hepatic dysfunction. Estrone was significantly elevated in both groups of patients. Estradiol concentrations were above normal in 10 patients with hepatocellular carcinoma and in 11 with cirrhosis only. All patients had normal concentrations of estriol. There were no statistical differences between the two groups in either individual or total estrogen levels (estrone 0.05 less than p less than 0.1). Eight of the patients with hepatocellular carcinoma and 5 of the cirrhotics had lower testosterone levels than normal, but this difference was not significant. However, the estrone to testosterone ratios were significantly higher in the hepatocellular carcinoma group than in the cirrhosis group (p less than 0.05). The present study seems to indicate that hyperestrogenemia commonly seen in male patients with liver cirrhosis may play some role in hepatic carcinogenesis of cirrhotic livers. Further studies are needed to determine if the estrone to testosterone ratio is implicated in hepatocarcinogenesis in cirrhotic men. PMID- 2981754 TI - Posttransfusion non-A, non-B hepatitis in chimpanzees. Physicochemical evidence that the tubule-forming agent is a small, enveloped virus. AB - Posttransfusion non-A, non-B hepatitis associated with the formation of hepatocyte cytoplasmic tubules was experimentally transmitted to chimpanzees by intravenous inoculation of a proven-infectious plasma that had been pelleted and microfiltrated, or purified by a combination of pelleting and rate-zonal banding. The results of these studies indicate that a factor VIII-derived non-A, non-B tubule-forming agent will pass through an 80-nm membrane filter and that it can be recovered from infected plasma by use of a purification procedure that assumes the non-A, non-B tubule-forming agent is a small, enveloped virus. Our findings, in combination with the known sensitivity of the non-A, non-B tubule-forming agent to chloroform and its apparent lack of nucleic acid homology with hepatitis B virus, further suggest that at least one etiologic agent of human posttransfusion non-A, non-B hepatitis may be a small, enveloped RNA virus. PMID- 2981755 TI - Watery diarrhea-hypokalemia-achlorhydria syndrome and carcinoma of the esophagus. AB - The watery diarrhea-hypokalemia-achlorhydria syndrome associated with ectopic secretion of vasoactive intestinal peptide has only been conclusively documented with tumors originating in the pancreas or sympathetic chain. We report here the case of a 50-yr-old woman who developed this syndrome 3 wk after an apparently effective course of radiotherapy for an obstructing, mixed-cell carcinoma of the esophagus. High concentrations of vasoactive intestinal peptide were found in plasma (100-200 pmol/L; normal less than 20 pmol/L) and in the metastatic skin nodules (750 pmol/g) that later developed and that contained one of the two cell types from the original tumor. Stool volumes reached a plateau of 15-20 L/day, and potassium requirements were greater than 1000 mmol/day. Symptoms failed to respond to any of the regimens previously described as effective in this syndrome. After 14 wk of massive fecal fluid and electrolyte losses, symptoms resolved dramatically with the first dose of 5-fluorouracil. Plasma vasoactive intestinal peptide concentration returned to normal, where it remained despite subsequent evidence of renewed tumor spread. This case illustrates the unpredictability of the response of this syndrome to medical treatment, and suggests that vasoactive intestinal peptide secretion may occur in a wider range of tumors than has so far been described. PMID- 2981756 TI - Mismatch repair mutations of Escherichia coli K12 enhance transposon excision. AB - Excision of the prokaryotic transposon Tn10 is a host-mediated process that occurs in the absence of recA function or any transposon-encoded functions. To determine which host functions might play a role in transposon excision, we have isolated 40 mutants of E. coli K12, designated tex, which increase the frequency of Tn10 precise excision. Three of these mutations (texA) have been shown to qualitatively alter RecBC function. We show that 21 additional tex mutations with a mutator phenotype map to five genes previously identified as components of a methylation-directed pathway for repair of base pair mismatches: uvrD, mutH, mutL, mutS and dam. Previously identified alleles of these genes also have a Tex phenotype.--Several other E. coli mutations affecting related functions have been analyzed for their effects on Tn10 excision. Other mutations affecting the frequency of spontaneous mutations (mutT, polA, ung), different excision repair pathways (uvrA, uvrB) or the state of DNA methylation (dcm) have no effect on Tn10 excision. Mutations ssb-113 and mutD5, however, do increase Tn10 excision.- The products of the mismatch correction genes probably function in a coordinated way during DNA repair in vivo. Thus, mutations in these genes might also enhance transposon excision by a single general mechanism. Alternatively, since mutations in each gene have qualitatively and quantitatively different effects on transposon excision, defects in different mismatch repair genes may enhance excision by different mechanisms. PMID- 2981757 TI - The gene structures of spontaneous mutations affecting a Caenorhabditis elegans myosin heavy chain gene. AB - We have isolated spontaneous mutations affecting the unc-54 major myosin heavy chain gene of Caenorhabditis elegans (variety Bristol). Spontaneous unc-54 mutants occur in C. elegans populations at a frequency of approximately 3 X 10( 7). We have studied the gene structure of 65 independent unc-54 mutations using filter-transfer hybridization techniques. Most unc-54 mutations (50 of 65) exhibit no abnormalities detected with these techniques; these mutations are small lesions affecting less than 100 base pairs. Approximately 17% of the mutations (11 of 65) are simple deletions, ranging in size from less than 100 base pairs to greater than 17 kilobases. One isolate contains two separate deletions, each of which affects unc-54. Two mutants contain tandem genetic duplications that include a portion of unc-54 and extend 8-10 kilobases beyond the 5' terminus of the mRNA. Conspicuously absent from our collection of spontaneous unc-54 mutations are any resulting from insertion of transposable genetic elements. Such mutants, if they occur, must arise at a frequency of less than 5 X 10(-9). PMID- 2981758 TI - Dysgenesis-induced instability of rosy locus transformation in Drosophila melanogaster: analysis of excision events and the selective recovery of control element deletions. AB - Utilizing the method of P-M hybrid dysgenesis-mediated gene transfer to insert rosy locus DNA into various chromosomal locations, we recovered a transformed strain that carries an ry+ transposon inserted in or near the scalloped locus in polytene section 13F on the X chromosome. The resultant product, when stabilized, behaves as a homozygous and hemizygous viable and fertile extreme scalloped allele associated with wild-type expression of the rosy locus. We have labeled this allele, sdry+. This allele has been destabilized by subsequent P-M hybrid dysgenesis, and mutations were recovered that exhibit alterations in the rosy and/or scalloped phenotypes. Representative samples of all phenotypic classes have been characterized by Southern blot analyses of restricted DNA. The most common events are excisions of DNA wholly internal to the transposon and representing sections of rosy DNA. In addition to loss of rosy locus function, such excisions affect the scalloped locus expression.--A second dysgenesis experiment was carried out involving an ry+ transposon inserted in polytene section 16D on the X chromosome. A minimal estimate of the relative frequency of imprecise excisions, determined in this experiment is 75%.--A successful pilot experiment is described that utilizes dysgenic perturbation of the sdry+ allele to select for small deletions of the 5' noncoding region of the rosy locus. PMID- 2981759 TI - Brown adipose tissue heat production in heat acclimated and perchlorate treated rats. AB - The noradrenaline-induced energy dissipation rate was measured with a direct microcalorimeter in brown adipose tissue taken from rats acclimated to 34 degrees C (HA), perchlorate treated (PC) and heat acclimated-perchlorate treated (HAPC). The response to 10(-7) M NA was reduced by 45%, 47% and 86% in HA, PC and HAPC groups, respectively, as compared to a control group kept at 24 degrees C. In the same groups, the response to 10(-6) M NA was reduced by 34%, 7% and 64%, respectively. The specific activity of the soluble alpha-glycerophosphate dehydrogenase in brown fat from HA rats was reduced by 50%, whereas it was not altered in the PC animals. It is concluded that the sensitivity to noradrenaline of the brown adipose tissue thermogenic mechanisms is decreased in hypothyroidism, and that the acclimation temperature and the thyroid status per se each have a different influence on brown adipose tissue function. PMID- 2981760 TI - Triiodothyronine increases serum angiotensin converting enzyme. AB - To determine whether elevated thyroid hormone is responsible for increased serum angiotensin converting enzyme in hyperthyroidism, 5 to 40 micrograms of 3,5,3' triiodo-L-thyronine was administered orally and subcutaneously to female Swiss Webster mice. Serum angiotensin converting enzyme was significantly increased in all animals given triiodothyronine compared to controls. Lung and kidney enzymes were moderately reduced in specific activity but unchanged in total activity due to increase in size of these organs. The results indicate that in hyperthyroidism, elevated thyroid hormone per se rather than the disease of the thyroid is responsible for elevated serum angiotensin converting enzyme. PMID- 2981761 TI - Nuclear thyroxine and triiodothyronine binding in mononuclear cells in dependence of age. AB - Nuclear binding of thyroxine (T4) and triiodothyronine (T3) in mononuclear blood cells was investigated in 12 young (age 16-30 years) healthy subjects (group A), in 12 middle-aged (age 31-60 years) healthy subjects (group B) and in 12 elderly (61-90 years) healthy subjects. Serum free T3 was depressed in group C as compared to the younger age groups, whereas serum free T4 and TSH did not differ between the groups. Maximal specific nuclear binding capacity for both T4 and T3 decreased with increasing age, T4 group A: 1.2 fmol T4/100 micrograms DNA, group B: 1.2 fmol T4/100 micrograms DNA, group C: 0.7 fmol T4/100 micrograms DNA; T3 group A: 1.7 fmol T3/100 micrograms DNA, group B: 1.0 fmol T3/100 micrograms DNA, group C: 0.9 fmol T3/100 micrograms DNA. The equilibrium association constant (Ka) for T4 increased with age, group A: Ka = 3.3 X 10(9) l/mol, group B: Ka = 3.2 X 10(9) l/mol, group C: Ka = 6.4 X 10(9) l/mol, whereas Ka for nuclear binding of T3 decreased with age group A: Ka = 3.9 X 10(9) l/mol, group B: Ka = 5.9 X 10(9) l/mol, group C: Ka = 1.8 X 10(9) l/mol. We conclude that, whereas the opposite variations of nuclear capacity and binding affinity for T4 tend to preserve the nuclear T4 concentration, the nuclear T3 concentration definitely decreases with age. The unaltered serum levels of TSH suggest that the decrease of both serum levels of free T3 and the nuclear T3 concentration might represent physiologically changes in old age. PMID- 2981762 TI - Effect of streptozotocin-diabetes and insulin treatment on regulation of Leydig cell function in the rat. AB - The present study was performed to further clarify the possible role played by insulin deficiency on the steroidogenic capacity of the rat testis. Sprague Dawley rats weighing 250-300 g were used in all experiments. Diabetes was induced by i.p. injection (40 mg/kg b.w.) of streptozotocin and was monitored at 2-day intervals by measuring body weight and serum glucose, glucosuria and ketonuria levels. The effect of insulin therapy on pituitary LH content and plasma LH concentrations, as well as on the cyclic AMP level in interstitial cell incubation medium and plasma testosterone concentrations, was measured 30 days after the induction of diabetes by radioimmunoassay. Streptozotocin-induced diabetes resulted in significantly reduced pituitary LH (16%, P less than 0.025) and plasma LH (34%, P less than 0.02); insulin treatment completely restored these levels. Similarly, the cyclic AMP content of interstitial cell incubation medium and the plasma testosterone concentrations were dramatically decreased in the diabetic state (50%, P less than 0.005 and 63%, P less than 0.025, respectively) and combined treatment with insulin plus hCG appeared slightly more effective than treatment with either of these hormones alone, suggesting a possible synergistic action. It is concluded that decreased testicular steroidogenesis in the diabetic rat may represent, at least in part, a direct consequence of insulin deficiency at the hypothalamic and/or pituitary levels. However, our findings would also be consistent with other reports suggesting that insulin may play a direct role in the rat testis. PMID- 2981763 TI - Prevalence of HBeAg and anti-HBe in chronic active hepatitis, cirrhosis and primary hepatocellular carcinoma in Korea. AB - The prevalence of HBeAg was studied in Korean patients with chronic active hepatitis, cirrhosis and primary hepatocellular carcinoma. There is an independent association of HBeAg prevalence with age and diagnosis. These findings are discussed in terms of a developmental model for primary hepatocellular carcinoma which could explain the lower frequency of HBeAg in the cancer cases. PMID- 2981764 TI - Liver morphology in southern African blacks with hepatocellular carcinoma: a study within the urban environment. AB - Hepatocellular carcinoma has a lower prevalence and presents at a later age in urban Blacks than in rural Blacks. These differences have previously been shown not to be attributable to differences in serum hepatitis B virus markers. In the present study, the average age of patients with hepatocellular carcinoma in a developing urban Black population is shown to have risen from 38.9 to 56.5 years (p less than 0.0001) over a 20-year interval, while the prevalence of co-existing cirrhosis has declined from 66 to 44% (p less than 0.05) and tissue HBsAg positivity has fallen from 44 to 17.7% (p = 0.002). The lower prevalence of tissue HBsAg in the recent patients may be explained by their older age. Macronodular cirrhosis was present in 56% of cases in the earlier period but declined to 18.9% in the later period, with micronodular cirrhosis becoming the dominant nontumor pathology (p = 0.002). Liver damage attributable to the abuse of alcohol is now found in more than half of the cases (48/90) of hepatocellular carcinoma occurring in this population. The remainder show no changes (12 cases) or show macronodular or incomplete septal cirrhosis (30 cases), presumed to be of viral origin. The latter cases are more likely to have serum markers of current hepatitis B virus infection than those with evidence of alcohol abuse. We conclude that alcohol is increasing in importance as an etiologic association of hepatocellular carcinoma in urban South African Blacks. At the same time, the prevalence of macronodular cirrhosis (and of cirrhosis as a whole) in urban patients with this tumor has declined. The reason for this decline is not known. PMID- 2981765 TI - Radioimmunoimaging of human small cell lung carcinoma with I-131 tumor specific monoclonal antibody. AB - In this study, an IgM monoclonal antibody (MAb600D11) directed against human small cell lung cancer (NCI-H69) was radiolabeled with iodine-131, and the biodistribution and image quality of the radiolabeled antibody was evaluated. Radiolabeling was achieved in a solid-phase system consisting of 1,3,4,6 tetrachloro-3a,6a-diphenylglycoluril. Labeling efficiencies and protein purification were accomplished using gel exclusion chromatography while radioimmunoreactivity was determined using a solid-phase radioimmunoassay procedure. The biodistribution of I-131-labeled MAbs was determined in Sprague Dawley rats up to 7 days after injection. Highest organ concentrations were observed in liver (3.91 +/- 0.47 (SD) and 0.17 +/- 0.04 (SD) mean percent injected dose at 1-7 days after injections) and in thyroid (5.33 +/- 0.71 (SD) and 5.32 +/- 2.01 (SD) mean percent injected dose at 1-7 days after injection). Nude mice, bearing either a small cell lung tumor (NCI-H69) or a nonspecific tumor (adenocarcinoma), were injected with 400-800 microCi of I-131 labeled monoclonal antibody. Optimum tumor visualization was observed 2-4 days after injection with tumor concentrations as high as 10.4% of the initial injected dose. The results demonstrated that radioimmunoimaging of human small cell lung carcinoma was feasible with the tumor-specific IgM I-131-labeled MAb. PMID- 2981766 TI - Analysis of lymphopoietic stem cells with a monoclonal antibody to the rat transferrin receptor. AB - A mouse monoclonal IgG2a antibody, designated MRC OX-26, is shown to be specific for the rat transferrin receptor, but does not block transferrin binding. The antibody labelled a myeloma, three leukaemia cell lines and normal dividing cells of various types, but also bound to a number of nondividing normal tissues. No labelling of lymphopoietic stem cells could be detected, even though approximately 25% of bone marrow and over 95% of fetal liver cells were clearly labelled. PMID- 2981767 TI - Activation of mouse macrophages causes no change in expression and function of phorbol diesters' receptors, but is accompanied by alterations in the activity and kinetic parameters of NADPH oxidase. AB - Mouse peritoneal macrophages activated in vivo by the injection of Corynebacterium parvum release larger amounts of superoxide anion (O2-) than macrophages from control mice when stimulated with phorbol myristate acetate (PMA). The biochemical bases for this enhanced response of activated macrophages have been investigated by studying the expression and function of receptors for the stimulant, and the activity of the enzyme NADPH oxidase which is responsible for the production of O2- in leucocytes. Studies of binding of phorbol dibutyrate, an agent closely related to PMA, showed that the affinity constants (Kds) and the number of binding sites were the same in resident and activated peritoneal macrophages. The activity of the NADPH oxidase was, however, different in the two macrophage populations which differ in their capacity to release O2-. NADPH oxidase activity was studied in macrophage monolayers after lysis with deoxycholate. The main features of this activity were as follows: stimulation of macrophages with PMA or zymosan caused an increase in NADPH-dependent O2- production; NADPH oxidase activity in the lysates followed the same dose-response curve for different concentrations of PMA as O2- release by intact macrophages; O2- release by intact macrophages could be fully accounted for by NADPH-dependent O2- production by macrophage lysates; activity was strictly substrate-specific, in that NADH could not substitute for NADPH; after stimulation with PMA or zymosan, NADPH oxidase activity was higher in lysates of C. parvum-activated macrophages than in lysates of resident macrophages; NADPH oxidase activities of activated and resident macrophages differed markedly in their kinetic parameters. The NADPH oxidase of macrophages activated by C. parvum or trehalose dimycolate of mycobacterial origin displayed a five to seven times lower Km compared to the enzyme in resident macrophages. PMID- 2981768 TI - Characterization of the HLA-A2.2 subtype: T cell evidence for further heterogeneity. AB - Five blood donors were identified whose HLA-A2 is different from the common HLA A2. Their A2 molecule (A2.2) had a more basic isoelectric point than normal A2 (A2.1). Cytotoxic T lymphocytes (CTL) restricted by HLA-A2.1, specific for influenza A and Epstein-Barr viruses, failed to lyse virus-infected target cells with HLA-A2.2. Identical patterns were obtained with both viruses. CTL from four of the A2.2-positive donors recognized target cells prepared from others in the group that shared only the HLA-A2.2 antigen. The A2.2 antigen from one donor seemed to be different in that target cells were not recognized by CTL from donors with the normal A2.1 nor with basic A2.2. There seems, therefore, to be heterogeneity within the HLA-A2.2 subtype. PMID- 2981770 TI - The gene encoding the Ia-associated invariant chain is located on chromosome 18 in the mouse. AB - The chromosomal assignment of the gene encoding the invariant (Ii) chain associated with the mouse immune response antigens (Ia) was determined by Southern blot analysis of DNA from a panel of mouse X Chinese hamster somatic cell hybrids cleaved with Hind III or Eco RI. Using a mouse li cDNA as a hybridization probe, we localized the gene coding for the invariant chain to mouse chromosome 18. PMID- 2981771 TI - Candidacidal activity of myeloperoxidase: therapeutic influence of the enzyme in vivo. AB - Mice were injected intravenously with Candida albicans blastospores to establish chronic renal infection. Seventy-five percent of the animals inoculated with 10(6) blastospores died as a consequence of infection during the subsequent 60 days of observation. Intraperitoneal administration of 10 mU of partially purified human myeloperoxidase 1 day after injection of the pathogen increased survival of the mice from 25 to 80% over this time period. Administration of myeloperoxidase complexed with soluble C. albicans cell wall mannan abrogated the protective influence of the enzyme. These results demonstrate that exogenous myeloperoxidase has a therapeutic influence on murine renal candidiasis and suggest that exogenous myeloperoxidase may also be effective in the treatment of certain forms of candidiasis in humans. These results also demonstrate the important role of the mannan-binding function of myeloperoxidase for effective treatment of candidiasis and suggest a mechanism of inhibition of the candidacidal effect of free enzyme in vivo by mannan accumulating in tissue fluids. PMID- 2981769 TI - DNA polymorphism of the C2 and factor B genes. Detection of a restriction fragment length polymorphism which subdivides haplotypes carrying the C2C and factor B F alleles. AB - Factor B and the second component of complement (C2) in man are encoded within the major histocompatibility complex by single loci that are less than 1 kb apart. A 2.3 kb factor B-specific cDNA probe has been used to examine, by Southern blot analysis, the genomic DNA of individuals typed for C2 and factor B by protein electrophoresis. We have identified a restriction fragment length polymorphism using the endonuclease Taq I, which subdivides haplotypes carrying both the common variant of C2 (C2C) and the fast (F) variant of factor B. This DNA polymorphism has been mapped to lie in the C2 gene and represents a new genetic marker not defined by protein electrophoresis. This polymorphism may serve as a useful marker in the genetic analysis of diseases that are related to the major histocompatibility complex. PMID- 2981772 TI - A conjugative transposon (Tn919) in Streptococcus sanguis. AB - Streptococcus sanguis FC1, originally isolated from dental plaque, was found to be naturally resistant to tetracycline. Although no plasmid DNA could be detected, tetracycline resistance was transferable in filter matings to Streptococcus faecalis FA2-2. Again, no plasmid DNA was detectable in transconjugants, and the latter could donate tetracycline resistance to S. faecalis, S. sanguis, and Streptococcus lactis. The tetracycline resistance element was able to transpose to several sites on the S. faecalis hemolysin plasmid pAD1 and in each case resulted in a 15-kilobase insert. DNA filter blot hybridization studies showed that the element bears significant homology with the conjugative transposon Tn916. Designated Tn919, it was cloned into an Escherichia coli plasmid vector (pGL101) and, as has been shown for Tn916, excised readily in the absence of selective pressure. PMID- 2981773 TI - Influence of nutrient limitation and low pH on serogroup B Neisseria meningitidis capsular polysaccharide levels: correlation with virulence for mice. AB - Neisseria meningitidis strain M986, which possesses a polyanionic sialic acid capsular polysaccharide, was resistant to the bactericidal effects of normal rabbit serum, but sensitive when immune serum and complement were present. An isogenic strain PRM102, deficient in the ability to produce capsular polysaccharide, was sensitive to normal serum. Strain M986, when grown under conditions of low pH or nutrient limitation, synthesized increased levels of capsular polysaccharide. This was accompanied by an increase in cell surface hydrophilicity and virulence for mice. Cells grown in low-pH, iron-limited medium synthesized the highest concentration of polysaccharide and exhibited the highest cell surface hydrophilicity and virulence among the cases examined. The increase in capsular polysaccharide was partly explained by a decrease in the specific activity of a membrane-bound cytidine monophosphate-N acetylneuraminic acid hydrolase. The results suggest that conditions of nutrient limitation and low pH exert profound effects on the physicochemical nature of the meningococcal cell surface which, in turn, cumulate in enhanced virulence of this organism for mice. PMID- 2981775 TI - Occurrence of IgA subclasses (IgA1 and IgA2) in the human nervous system. Correlation with disease. AB - The occurrence of IgA subclasses in pathological conditions of the nervous system was studied by means of monoclonal antibodies and an indirect immunofluorescence technique. IgA1- and/or IgA2-positive lymphoid (plasma) cells were found in demyelinating diseases comprising multiple sclerosis, Guillain-Barre syndrome, and adrenoleukodystrophy, in various inflammatory diseases, and in tumors, some of which exhibited labeling of tumor cells. Demyelinating and inflammatory diseases with chronic course displayed some prevalence of IgA2-, and tumors some prevalence of IgA1-positive cells. This is the first demonstration of IgA1 and IgA2 in the nervous system. PMID- 2981774 TI - Relationship between extracellular stimulation of intracellular killing and oxygen-dependent microbicidal systems of monocytes. AB - Human monocytes require serum components immunoglobulin G, C3/C3b, and B/Bb to exert optimal microbicidal action against ingested microorganisms. The present study was performed to find out whether these factors act by enhancing oxygen dependent antimicrobial mechanisms. Serum enhanced oxygen consumption and superoxide production by monocytes before phagocytosis, but did not further increase these processes in monocytes that had recently ingested bacteria. Furthermore, serum did not boost iodination during intracellular killing by monocytes. Phorbol myristate acetate, N-formyl-methyonyl-leucyl-phenylalanine, concanavalin A, and concanavalin A-Sephadex all stimulated the conversion of O2 to H2O2 by monocytes, but only concanavalin A augmented intracellular killing. Reactive oxygen intermediates generated by cell-free enzymes (xanthine oxidase or glucose oxidase) in concentrations comparable to those accumulating extracellularly during incubation of monocytes containing bacteria with phorbol myristate acetate did not promote intracellular killing. The presence of catalase during phagocytosis inhibited killing, but had no effect on killing in the postphagocytic state. Monocytes deprived of glucose for 24 h showed markedly impaired O2 consumption, O2- generation, and bacterial killing; all of these effects were rapidly reversed by restoration of glucose. It is concluded that both an intact respiratory burst and extracellular serum factors are necessary for optimal killing of intracellular Staphylococcus aureus by human monocytes. Serum does not appear to act by enhancing the respiratory burst, but rather to have a separate, synergistic role, the biochemical basis of which is unknown. PMID- 2981776 TI - Aggregating and prostanoid-releasing effects of platelet-activating factor and leukotrienes on human polymorphonuclear leukocytes and platelets. AB - Aggregating and prostanoid-releasing properties of the inflammatory mediators, platelet-activating factor (PAF) and leukotrienes B4, C4 and D4 were studied in human polymorphonuclear leukocytes (PMNL) and in human platelet-rich plasma. Leukotriene B4 (LTB4) and PAF both induced a reversible aggregation of human PMNL with concomitant stimulation of PGE2 formation, whereas LTC4 had no effect on human PMNL. Arachidonic acid (AA) caused an irreversible aggregation of PMNL which was accompanied by formation of both PGE2 and TXB2. Inhibition of TXB2 synthesis by indomethacin or by OKY-1581, a thromboxane synthetase inhibitor, had no effect on the PMNL aggregation induced by LTB4, PAF or AA. Leukotrienes B4, C4 and D4 caused neither aggregation nor TXB2 release in human platelet-rich plasma. PAF, on the other hand, induced a dose-dependent, reversible platelet aggregation which was not accompanied by TXB2 formation nor inhibited by OKY-1581. The present study indicates that in addition to inducing PMNL aggregation, LTB4 is capable of releasing arachidonate metabolites from human PMNL but not from human platelets. Also the responses of PMNL and platelets to PAF seem to differ as the PAF-induced PMNL aggregation was accompanied by increased prostanoid formation whereas the PAF-induced reversible platelet aggregation was obviously independent from arachidonate metabolism. PMID- 2981777 TI - Effect of 24 antimicrobial drugs on polymorphonuclear leukocyte adherence. AB - The effects of 24 antimicrobial agents on the adherence of human neutrophils to nylon fibre microcolumns were studied. Neutrophil adherence was found to be remarkably resistant to antimicrobial agents. No effect was observed with penicillin G, nafcillin, cephalothin, vancomycin, bacitracin, tetracycline, minocycline, doxycycline, chloramphenicol, erythromycin, lincomycin, neomycin, streptomycin, acetylspiramycin, 5-fluorocytosine, sulfisoxazole, tinidazole and primaquine. Only 2 enhanced adherence - oxytetracycline and rifampicin - and only 4 suppressed adherence - colistin, polymyxin B, quinine, chloroquine - and then only at high concentrations. PMID- 2981778 TI - A prospective study of association of herpes simplex virus and human papillomavirus infection with cervical neoplasia in women exposed to diethylstilbestrol in utero. AB - One thousand one hundred thirty-four women, enrolled in a prospective study of the development of cervical neoplasia in women exposed to diethylstilbestrol (DES) in utero, were examined at least once a year. Twenty-three women developed cervical intraepithelial neoplasia (CIN) of different degrees during the 7-year follow-up period and were matched with women who did not develop CIN during the study period. Serum samples were obtained at the time of entry into the study and at the time of diagnosis of neoplasia. In the control group, a second serum sample was obtained close to the time when CIN was diagnosed in the other group of women. The sera were studied for antibodies to both types of herpes simplex virus (HSV) by microneutralization assay and by a radioimmunoassay using, as antigens, glycoproteins VP123 for HSV type 1 and VP119 for HSV type 2. Women who developed CIN had, in the enrollment specimen which preceded the neoplasia, a higher rate of antibodies to HSV-1 than did the matched controls (22%), while the two groups did not differ in frequency of antibodies to HSV-2 (9%). Similar differences were observed in the sample drawn at the time of diagnosis of CIN. Women who later developed CIN and had, at the time of entry into the study, cervical epithelial changes associated with DES exposure, were infected with HSV 1 at a higher rate than women who remained free of neoplasia or women with CIN but without DES-associated cervical epithelial changes. Morphologic evidence of associated papillomavirus (HPV) infection was observed in 74% of CIN cases and no such observation was made in biopsy specimens taken prior to development of CIN. Morphologic evidence of HPV infection was found in a women of the control group having squamous metaplasia. HPV structural antigens were found in 3 of 10 CIN I and CIN II cases, and none in the CIN III specimens. The possible role of the HSV and HPV infection in the pathogenesis of cervical neoplasia is discussed. PMID- 2981780 TI - A re-examination of the Epstein-Barr virus carrier state in healthy seropositive individuals. AB - A panel of 24 healthy seropositive donors have been followed prospectively over a period of 15 months and monitored (1) for the level of EB virus shedding in the throat by means of a sensitive cord-blood transformation assay; (2) for the level of virus-infected B cells in the blood via a new in vitro protocol where "spontaneous transformation" can be seen to titrate against input cell number; (3) for anti-EB viral antibody titres and (4) for the prevailing level of virus specific memory T cells in the circulation. Six donors shed easily detectable levels of EB virus into throat washings on every occasion of testing, whilst 16 other donors shed lower levels of virus detectable in throat washings on a majority (10 donors) or on a minority (6 donors) of test occasions; only 2/24 donors gave no evidence of virus shedding at any time. There was a direct relationship between the EB virus shedder status of an individual (i.e., the level of virus replication in the pharynx) and the number of infected B cells present in the circulation. These results indicate that chronic, usually low grade, replication of the virus at some permissive site in the oro- and/or naso pharynx is very often a stable accompaniment of the asymptomatic EB virus carrier state, and may indeed be essential for the long-term maintenance of that state. PMID- 2981779 TI - Establishment of the Amsalem T-cell line from a patient with acute lymphoblastic leukemia. Expression of E-receptor-associated antigens in cells incapable of forming E-rosettes. AB - A new T-cell line (Amsalem) was established from the peripheral blood of a patient with pre-T leukemia. Amsalem cells are unique in that they possess antigenic determinants associated with the E-receptor, yet fail to form rosettes with sheep red blood cells (SRBC). Amsalem cells were found to possess morphological and cytochemical features characteristic of T-lymphocytes, and were sensitive to the cytotoxic effect of rabbit antisera specific for T-cell antigens. In immunofluorescent tests with monoclonal antibodies, Amsalem cells showed a strong reactivity with the OKT-11A and A-22 antibodies, specific for the E-receptor. The cells were reactive with OKT-4 and showed a very weak reactivity with OKT-6 and OKT-8. No reactivity was found with the OKT-3, Leu 7, Leu 11, and OKM1 antibodies. Amsalem cells failed to form rosettes with SRBC; however, mouse anti-Amsalem serum inhibited the formation of E-rosettes. It is concluded that the Amsalem cell line is a line of pre-T leukemia cells characterized by a dissociation between its inability to form E-rosettes and the presence of antigenic constituents of the E-receptor. PMID- 2981781 TI - In vitro analysis of the Epstein-Barr virus: host balance in long-term renal allograft recipients. AB - Four indices of the EB virus carrier state, for which quantitative in vitro assays now exist, have been monitored in 55 renal allograft recipients under long term immunosuppression, each patient being tested on a single occasion. By comparison with parallel data from healthy control donors, the results indicate the extent to which virus replication in the throat and virus-infected B cells in the blood are increased as a result of immunosuppression; the concordance between these two independent indices of the level of EB virus infection in vivo, first noted with healthy donors, was again observed within this large group of patients. Immunosuppression also leads to an impairment of virus-specific memory T-cell responsiveness and to an increase in anti-viral antibody titres, but the results show that the level of virus infection prevailing in any one individual cannot be inferred directly from these immunological indices of the virus:host balance. In allograft patients on stable levels of immunosuppression, virus and host appear to establish a new equilibrium. Limited prospective studies suggest that the position of this new equilibrium depends critically upon the virus:host balance prevailing in the same individuals before immunosuppression began. This may be an important consideration in identifying patients for whom immunosuppression may carry a particularly high risk of developing EB virus genome-positive lymphoma. PMID- 2981782 TI - Relationship between serum and histochemical markers for hepatitis B virus and rate of viral integration in hepatocellular carcinomas in Japan. AB - The relationship between serological and histochemical markers of hepatitis B virus (HBV) and viral integration in hepatocellular carcinomas (HCCs) in Japan was investigated. Special attention was paid to the exclusion of false-positive results due to bacterial contamination of autopsy materials. Of 85 patients with HCC, 23 were positive for serum HBsAg, 18 were positive for serum HBV antibodies and the remaining 44 were negative for serum HBV markers. Among the 23 HCCs from HBsAg carriers, 19 (82.6%) had integrated HBV DNA in the tumor DNA. In contrast, of the 18 HCCs from HBV antibody-positive patients, only one was positive for HBV DNA integration; this particular HCC was from a long-term HBsAg carrier who became a non-carrier by seroconversion at a later age. Of 44 HCCs from patients negative for serum HBV markers, 3 had integrated HBV DNA. Of these 3 cases, 2 were, however, histochemically positive for HBsAg in non-cancerous portions of the liver. Thus, our present results show a high integration rate of HBV DNA in HCCs of carrier patients and an extremely low rate of viral integration in HCCs of non-carrier patients. The latter finding contrasts strongly with previous results gained from analysing materials from patients in Europe and Africa. No histological differences were apparent between HCCs with or without viral integration. PMID- 2981783 TI - Herpes simplex virus type 2 and human papillomavirus type 16 in cervicitis, dysplasia and invasive cervical carcinoma. AB - Sera and biopsies of cervical lesions from 55 women with benign or malignant disease were analyzed for evidence of infection with herpes simplex virus type 2 (HSV-2) or human papillomavirus (HPV). In addition, information regarding known risk factors for cervical cancer was obtained by interview. The sera were tested for HSV-2 antibodies and the biopsies were tested for HPV or HSV DNA sequences by Southern blot hybridization. HSV-2 sequences were detected in 2/13 (15%) invasive neoplasms and in 1/12 (7%) benign lesions. Under non-stringent conditions of hybridization, reactions with HPV DNA were detected in biopsies of 2/17 (12%) inflammatory lesions, 6/12 (50%) intraepithelial neoplasms and 13/20 (65%) invasive neoplasms. All but one of the positive biopsies of invasive cancer, but only 4/11 biopsies of non-invasive lesions, contained HPV-16 DNA as determined by stringent hybridization conditions. Women with cervical cancer possessed the risk factors associated with the disease. Cigarette smoking and the presence of HPV-16 DNA were the most prominent risk factors. No evidence of an interaction between HSV-2 and HPV-16 was found among the cases of invasive cervical cancer. PMID- 2981784 TI - Infectious transmission of human T-cell leukemia virus to rabbits. AB - A rabbit lymphoid cell line (Ra-1) was established by co-cultivation with a human T-cell line (MT-2) carrying human T-cell leukemia virus (HTLV). The Ra-1 cell line is chromosomally male and is persistently infected with HTLV. Ra-1 cells, with or without mitomycin C treatment, were inoculated intravenously (i.v.) into 3 female rabbits. All 3 animals responded with the production of antibodies to HTLV antigens. Lymphocytes from one of these seroconverters were cultured in the presence of T-cell growth factor (TCGF) and HTLV particles were detected in the TCGF-grown lymphocytes which were chromosomally female. Co-cultivation of lymphocytes from the 2 other seroconverters with lymphocytes from 2 anti-HTLV negative healthy men gave rise to the establishment of an HTLV-producing T-cell line derived from each individual. Blood transfusion from one of the HTLV infected rabbits into 2 female rabbits also resulted in the seroconversion of both recipients. An HTLV-carrying lymphoid cell line (Ra-2) was established from one of the transfusion-related seroconverters. The Ra-2 cell line was initially TCGF-dependent but later became TCGF-independent. There results indicate that HTLV can be transmitted to rabbits. These animals may provide a suitable model system for studying the mode of transmission and pathogenicity of HTLV. PMID- 2981785 TI - A study of angiogenesis factors from five different sources using a radioimmunoassay. AB - The use of a radioimmunoassay (RIA) demonstrated that angiogenic factors from human and animal tumours, normal bovine retinas, myocardial infarcts, synovial fluid from patients with joint diseases and wound fluid shared common antigenic determinants. Values for angiogenesis factors expressed as microgram Walker tumour TAF/mg protein varied from 0.8 (wound fluid) to 207 (myocardial infarct). Activated macrophages produced an angiogenic factor which did not cross-react. Normal tissue extracts which were non-angiogenic by the chicken chorioallantoic membrane assay also failed to cross-react in the RIA. The antigenic similarity of angiogenic factors from such a wide variety of sources suggests that, in order to minimize therapeutic side effects, it would be best to use angiogenic agonists and antagonists locally. PMID- 2981786 TI - HSV-1 quantitation from rabbit neural tissues after epinephrine-induced reactivation. AB - Epinephrine iontophoresis into the eye can induce ocular herpes simplex virus type-1 (HSV-1) shedding with a high frequency from latently infected rabbits. The present study was designed to qualify and quantify infectious HSV-1 from neural tissues of latently infected rabbits after ocular epinephrine iontophoresis. Epinephrine iontophoresis was performed daily for 3 consecutive days on selected days during 220-227 days postinoculation. The induced ocular shedding was detected in the tear film with a frequency of 83% (10/12) within 72 hr after the initial iontophoresis. The rabbits were killed 24 hr after the last iontophoretic treatment, and the corneas and neural tissues were homogenized immediately. The cell-free supernatants were inoculated on primary rabbit kidney cell monolayers for qualitative assays of infectious virus and later titrated on CV-1 monolayers. The frequencies of the recovery of infectious HSV-1 from the cell-free homogenates were 0% of the corneas (0/12), 83% (10/12) from the superior cervical ganglion (SCG), 100% (12/12) from the trigeminal ganglion (TG), 42% (5/12) from the ophthalmic branch of the trigeminal nerve (TN), 8% (1/12) from the root entry zone of the trigeminal nerve into the brain-stem (REZ), and 0% (0/12) from the cerebellum. The authors conclude that epinephrine iontophoresis can reactivate the latent HSV-1 in neural tissues and infectious virus can be quantified from the cell-free homogenates. To the best of our knowledge, this is the first report to quantify HSV-1 with a high frequency from neural tissues following induced reactivation. PMID- 2981788 TI - On some properties of 222Rn short-lived decay products in air. AB - Simultaneous measurements were made of such properties as the fraction of charged and uncharged atoms, the balance of radioactive equilibrium between 222Rn and its daughters, and the concentration of aerosol particles and their mean radii in tunnel air. It became clear that the behavior of 222Rn decay products in tunnel air could be expressed well by equations based on a simple model, taking the following into account: the attachment of free atoms to aerosol particles, the deposition of radioactive particles on the tunnel wall, emission of alpha recoils from aerosol particles and the surface of the tunnel wall, and radioactive decay. In addition, the effective attachment coefficient of an observed RaA-atom was found to agree well with that calculated. The results obtained should facilitate in the future estimation of the relation between 222Rn daughters and the lung dose to the population. PMID- 2981787 TI - Further studies on the influence of chemical form and dose on absorptions of Np, Pu, Am and Cm from the gastrointestinal tracts of adult and neonatal rodents. AB - Absorption of isotopes of the actinide elements Np, Pu, Am and Cm from various organic media and/or in combination with plant or animal ligands or tissue is compared with their absorption from an inorganic nitrate medium. Gastrointestinal (GI) transport of 238Pu, 239Pu, 241Am and 244Cm at high concentrations from citrate medium by adult rats and/or mice was higher than from nitric acid medium. Neptunium-237 absorption, however, was not increased by citrate; probably because its oxidation state was reduced from 237Np(V) to 237Np(IV) by the medium and by the GI content. Increasing the mass of the 237Np dose resulted in increased absorption. Neither incorporation of 238Pu in rat liver nor retention of 238Pu oxide in rat lungs enhanced absorption when those Pu-containing tissues were administered intragastrically to either adult or neonatal rats. Ranking of GI absorption of the various forms of Pu gavaged in these studies suggests that transport is in the order: Pu citrate greater than Pu phytate greater than biologically incorporated Pu greater than Pu nitrate. PMID- 2981789 TI - Practical aspects of clinical trials: a review of a randomized, placebo controlled study in patients with heart failure. PMID- 2981790 TI - Analysis of local-regional relapses in patients with early breast cancers treated by excision and radiotherapy: experience of the Institut Gustave-Roussy. AB - Between 1970 and 1981, 436 patients with T1 and small T2 breast carcinoma were treated by tumor excision followed by radiotherapy at the Institut Gustave Roussy. The mean follow-up was 5 years, with 50% of patients followed 5 years. Twenty-four patients have experienced a local-regional (LR) relapse for an actuarial LR control rate of 93% at 5 years and 90% at 10 years. Potential prognostic factors for all 24 local-regional recurrences and for the subgroup with relapses in the breast were analyzed. A high Bloom grade and low Nominal Standard Dose (NSD) were significant prognostic factors for predicting LR relapse in both groups. Disease-free survival (from initial presentation) was not adversely affected by a solitary breast recurrence, when patients with successful salvage treatment were considered disease free. However, the group of patients with nodal or dermal recurrences had a much worse prognosis. This paper describes the natural history of breast cancer following a local-regional relapse in irradiated patients without mastectomy. Most importantly, we observed that breast relapses following radiotherapy become clinically apparent more slowly than chest wall failures after mastectomy, and if detected early, that these patients may be successfully retreated. PMID- 2981791 TI - Isolation of activated ras transforming genes from two patients with Hodgkin's disease. AB - Despite impressive advances in the clinical management of Hodgkin's disease, little is known about its cellular origin or the mechanism(s) of "Hodgkinogenesis." Recent findings that certain human cellular oncogenes can cause malignant transformation suggest that aberrant activation of these genes may play a role in carcinogenesis. To determine if such genes are operative in Hodgkin's cells, we isolated DNA from splenic nodules of three patients with nodular sclerosis Hodgkin's disease and tested its ability to transform mouse NIH 3T3 cells, the standard assay for oncogene-mediated malignant transformation. Transformed cells containing human DNA were obtained from two patients. DNA from these primary transformants yielded secondary transformants of NIH 3T3 fibroblasts; one also transformed normal mouse bone marrow macrophages, a cell type probably related to the Hodgkin's cell. When analyzed by Southern blot methods for homology with closed oncogene probes, the transforming genes from both patients had homology with N-ras. The homology and size of the restriction fragments were similar to those of transforming genes isolated from patients with acute nonlymphocytic leukemias. The presence of the same activated oncogene in tumor tissue from two different patients suggests that it may play an important role in Hodgkinogenesis. PMID- 2981792 TI - Target cells and thymus microenvironment in the pathogenesis of thymic lymphomas in C57BL/Ka mice. AB - In C57BL/Ka mice, the induction of thymic lymphomas either by inoculation of radiation leukemia virus (RadLV) or by a split dose irradiation requires complex cellular events: Target cells are found among the population of thymic subcapsular blast cells, or, alternatively, of marrow or spleen prothymocytes; Progression of target cells to lymphoma growth requires a multi-step process, which occurs only within thymic microenvironment; Target cells are rapidly induced as "preleukemic" cells; After inoculation of RadLV, the initial events occur when target cells are in close association with cells of a specialized component of thymic epithelium, i.e., the so-called "nurse cells"; The leukemogenic agents induce damages to the thymic microenvironment itself; Lymphoma prevention by marrow grafting after irradiation results from mechanisms still unknown which inhibit the progression of "preleukemic" cells to neoplastic growth. PMID- 2981793 TI - Patterns of thymocyte differentiation markers on virus and radiation induced lymphomas of C57BL/Ka mice. AB - To better understand the biology of tumorigenesis in virus and radiation lymphomas of C57Bl/Ka mice, we have examined the cell surface phenotypes of a large series of primary tumors induced by both agents. Data derived using flow cytometry and recently available monoclonal antibodies to thymocyte differentiation antigens supports three major conclusions. First, tumor cell populations are unimodal for staining with most antibodies and are probably of clonal origin. Second, many, but not all, tumor cells show surface phenotypes similar to those of previously defined subpopulations of normal thymocytes. Third, at the cell surface level, no major differences between virus- and radiation-induced lymphomas can be discerned. Our data thus further define the relationship between thymomas induced by these two agents. PMID- 2981794 TI - Effect of cold exposure on liver and muscle cAMP content and cAMP phosphodiesterase activity. AB - Adenosine 3',5'-cyclic monophosphate (cAMP) concentration and 3',5'-cyclic nucleotide phosphodiesterase (PDE) activity were measured in skeletal muscle, heart, and liver of rats exposed to 1, 3, 5, and 7 days of cold. Cyclic nucleotide concentration increased in fast-twitch red muscle at the same time that PDE activity was decreasing. Nucleotide concentration and enzyme activity of slow-twitch red muscle were not altered by the cold exposure. The PDE activity of fast-twitch white muscle was elevated approximately 50% above control after 1 and 3 days of cold exposure. By the 5th day in the cold, white muscle PDE activity had returned to control levels and remained there through the 7th day of experimentation. cAMP concentration in hearts of cold-exposed rats was significantly (P less than 0.01) elevated above control at all time points measured. Myocardial PDE activity was elevated above control (P less than 0.05) at 1 and 3 days of cold exposure but returned to control levels by the 5th day in the cold. Hepatic cAMP and PDE activity were elevated above control at all time points analyzed. These data suggest that changes in cyclic nucleotide metabolism play a role in attaining homeostasis during acute cold exposure. PMID- 2981795 TI - Isolation of a versatile Serratia marcescens mutant as a host and molecular cloning of the aspartase gene. AB - An extracellular nuclease-deficient, antibiotic-sensitive, and restrictionless mutant was isolated from the wild-type strain of Serratia marcescens Sr41 by four rounds of mutagenesis. The mutant was transformed efficiently with plasmid DNAs prepared from Escherichia coli and S. marcescens, and was used as a host for the cloning of the aspartase gene (aspA+) of S. marcescens. Cells carrying the cloned aspA+ gene on a multicopy plasmid produced ca. 39-fold more aspartase than did control cells, and the level of enzyme overproduction was in proportion to the copy number of the aspA+ recombinant plasmid. Aspartase was identified as a polypeptide of molecular weight 52,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 2981797 TI - Isolation and characterization of an Escherichia coli mutant lacking the cytochrome o terminal oxidase. AB - A respiration-deficient mutant of Escherichia coli has been isolated which is unable to grow aerobically on nonfermentable substrates such as succinate and lactate. Spectroscopic and immunological studies showed that this mutant lacks the cytochrome o terminal oxidase of the high aeration branch of the aerobic electron transport chain. This strain carries a mutation in a gene designated cyo which is cotransducible with the acrA locus. Mutations in cyo were obtained by mutagenizing a strain that was cyd and, thus, was lacking the cytochrome d terminal oxidase. Strain RG99, which carries both the cyd- and cyo- alleles, grows normally under anaerobic conditions in the presence of nitrate. Introduction of the cyd+ allele into the strain restores the respiration function of the strain, indicating that the cytochrome o branch of the respiratory chain is dispensable under normal laboratory growth conditions. PMID- 2981796 TI - Spiroplasma membrane lipids. AB - Membranes of six spiroplasma strains belonging to different Spiroplasma species and subgroups were isolated by a combination of osmotic lysis and sonication in the presence of EDTA to block endogenous phospholipase activity. Analysis of membrane lipids showed that in addition to free and esterified cholesterol the spiroplasmas incorporated exogenous phospholipids from the growth medium. Sphingomyelin was preferentially incorporated from phosphatidylcholine sphingomyelin vesicles or from the serum used to supplement the growth medium. Palmitate was incorporated better than oleate into membrane lipids synthesized by the organisms during growth. The major phospholipid synthesized by the spiroplasmas was phosphatidylglycerol. The positional distribution of the fatty acids in phosphatidylglycerol of Spiroplasma floricola resembled that found in Mycoplasma species, in which the saturated fatty acids prefer position 2 in the glycerol backbone and not position 1 as found in Acholeplasma species and elsewhere in nature. Electron paramagnetic resonance analysis of spin-labeled fatty acids incorporated into S. floricola membranes exhibited homogeneous single component spectra without immobilized regions. The S. floricola membranes were more rigid than those of Acholeplasma laidlawii and less rigid than those of Mycoplasma gallisepticum. PMID- 2981798 TI - Independence of cyclic AMP and relA gene stimulation of glycogen synthesis in intact Escherichia coli cells. AB - Previous studies from our laboratory established that in Escherichia coli, glycogen synthesis is regulated by both the relA gene, which mediates the stringent response, and by cyclic AMP. However, those studies raised the question of whether this dual regulatory system functions in an independent or a dependent manner. We show here that this regulation is independent, i.e., each regulatory process can express its action in the absence of the other. Triggering the stringent response by amino acid starvation increased glycogen synthesis even in mutants lacking the ability to synthesize cyclic AMP or lacking cyclic AMP receptor protein; and cyclic AMP addition stimulated glycogen synthesis in relA mutant strains. We also show that physiological concentrations of GTP inhibit ADP glucose synthetase (glucose-1-phosphate adenylyltransferase, EC 2.7.7.27), the rate-limiting enzyme of bacterial glycogen synthesis, in vitro. Because the stringent response is known to cause an abrupt decrease in the cellular level of GTP, modulation of ADP-glucose synthetase activity by this nucleotide could account for a substantial portion of the step-up in the cellular rate of glycogen synthesis observed when the stringent response is triggered. PMID- 2981800 TI - Release of respiratory control in Escherichia coli after bacteriophage adsorption: process independent of DNA injection. AB - Adsorption of phages T4, T5, and BF23 to previously starved Escherichia coli cells triggered the immediate release of respiratory control. A similar stimulation of respiration was induced after T4 ghost attachment, showing that this process was independent of the mechanism of DNA injection. Rather, this change in the respiratory rate was related to the transient depolarization of the cytoplasmic membrane also induced after phage and ghost adsorption. Both processes were suppressed by addition of ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid in the case of T4 (phage and ghosts) but not of T5 and BF23. The increase in respiratory rate observed after phage adsorption was of a magnitude similar to that induced by protonophores. Since other treatments that depolarize the membrane without a massive proton influx did not increase the rate of respiration of starved bacteria with the same efficiency, these results suggest that phage adsorption induced an entry of protons into the cell cytoplasm. PMID- 2981799 TI - Cloning of a developmentally regulated element from alkalophilic Bacillus subtilis DNA. AB - An alkalophilic Bacillus DNA bank cloned in an expression probe plasmid, pGR71, was screened for the presence of developmentally regulated genetic elements. A 508-base pair HindIII fragment isolated from this bank in plasmid pGR71-5 expressed plasmid-encoded chloramphenicol resistance only during the sporulation phase of a Bacillus subtilis host grown on Schaeffer medium. This developmentally regulated expression was altered in spo0E and spo0H mutants which had very low levels of chloramphenicol acetyltransferase activity relative to the wild type or other spo0 mutants. We determined the nucleotide sequence of the entire 508-base pair fragment and located the site of regulated transcription initiation by high resolution S1 nuclease mapping of the in vivo transcript. The deduced promoter sequences upstream from this start site were 5'C-G-A-A-T-C-A-T-G-A3' at -10 and 5' A-G-G-A-A-T-C3' at -35. This transcript was not detected in spo0E or spo0H mutants, indicating that the products of these genes control developmentally regulated chloramphenicol acetyltransferase expression at the level of transcription. PMID- 2981801 TI - A 2.2-kilobase repeated DNA segment is associated with DNA amplification in Streptomyces fradiae. AB - We have previously identified a 10.5-kilobase DNA sequence which is highly amplified and tandemly repeated in the mutant Streptomyces fradiae JS85. A library of DNA was prepared from S. fradiae T776, which does not contain amplified DNA. The library was screened by plaque hybridization to identify phage clones containing the unamplified 10.5-kilobase DNA sequence. Four phage isolates were identified which contained DNA homology to the amplified DNA sequence. This sequence was designated the amplifiable unit of DNA. None of the clones carried an entire amplifiable unit of DNA, and so overlapping regions were aligned to create a map of the entire region. Detailed restriction mapping identified a 2.2 kilobase direct repeat at the ends of the amplifiable unit of DNA. Analysis by Southern hybridization confirmed that the direct repeats were homologous to each other. The DNA of S. fradiae contained at least two additional copies of DNA that was homologous to the repeat sequence. PMID- 2981803 TI - Generation of a large, protonophore-sensitive proton motive force and pH difference in the acidophilic bacteria Thermoplasma acidophilum and Bacillus acidocaldarius. AB - The mechanism by which acidophilic bacteria generate and maintain their cytoplasmic pH close to neutrality was investigated. For this purpose we determined the components of proton motive force in the eubacterium Bacillus acidocaldarius and the archaebacterium Thermoplasma acidophilum. After correction for probe binding, the proton motive force of untreated cells was 190 to 240 mV between external pH 2 and 4. Anoxia diminished total proton motive force and the transmembrane pH difference by 60 to 80 mV. The protonophore 2,4-dinitrophenol abolished the total proton motive force almost completely and diminished the transmembrane pH difference by at least two units. However, even after correction for probe binding, protonophore-treated cells maintained a pH difference of approximately one unit. PMID- 2981802 TI - Leucine tRNA family of Escherichia coli: nucleotide sequence of the supP(Am) suppressor gene. AB - We describe the cloning and the DNA sequence of an amber suppressor allele of the Escherichia coli leuX (supP) gene. The suppressor allele codes for a tRNA with anticodon CUA, presumably derived by a single base change from a CAA anticodon. The mature coding sequence of the leuX gene is preceded by a putative Pribnow box sequence (TATAAT) and followed by a termination signal. The sequence of the leuX coded tRNA is compared with the sequences of the four remaining tRNALeu isoacceptors of E. coli and with two tRNALeu species from bacteriophage T4 and T5. The conserved nucleotides in these seven tRNAs recognized by E. coli leucyl tRNA synthetase are located mainly in the aminoacyl stem and in the D-stem/loop region. PMID- 2981804 TI - Stable inheritance of plasmid R1 requires two different loci. AB - The largest EcoRI fragment from plasmid R1 mediates a stability phenotype which is required to ensure the stable inheritance of this low-copy-number plasmid. When covalently linked to small, unstable R1 derivatives, this fragment makes the plasmids as stable as the wild-type R1 plasmid. A genetic analysis showed that two independently acting stabilization functions are encoded by this EcoRI fragment, both of which have the potential of partial stabilization of mini-R1 plasmids. The two loci are located at opposite ends of the fragment. Stabilization was also obtained by inserting these regions in unrelated, unstable plasmids from the p15 group. One of the two functions was very efficient in stabilizing such foreign replicons. Besides the stability phenotype, these genes exert incompatibility in an allele-specific manner. The stability functions do not seem to interfere seriously with the copy number of the plasmid. PMID- 2981805 TI - Molecular cloning of the phenylalanine ammonia lyase gene from Rhodosporidium toruloides in Escherichia coli K-12. AB - A genomic library of Rhodosporidium toruloides DNA was constructed in bacteriophage lambda 1059. Recombinant phage containing phenylalanine ammonia lyase (PAL) gene sequences were identified by using 32P-labeled cDNA to partially purified PAL mRNA. The PAL gene was subcloned on an 8.5-kilobase PstI DNA restriction fragment into pUC8 to generate the recombinant plasmid pHG2. A restriction map of the PAL gene, together with its flanking regions, was constructed. Northern hybridization analysis of R. toruloides RNA with a restriction fragment encoding part of the PAL gene indicates that PAL mRNA is 2.5 kilobases in length. A single-stranded DNA hybridization probe was constructed and used to quantitate PAL mRNA levels in R. toruloides grown under different physiological conditions. PAL mRNA levels paralleled changes in functional PAL mRNA and antigen. These data are consistent with control of PAL expression being at the level of transcription. PMID- 2981806 TI - Cloning of a small, acid-soluble spore protein gene from Bacillus subtilis and determination of its complete nucleotide sequence. AB - The first Bacillus subtilis small, acid-soluble spore protein (SASP) gene has been cloned by using previously cloned B. megaterium SASP genes as DNA-DNA hybridization probes. Determination of the DNA sequence of the B. subtilis SASP gene showed that it codes for a 72-residue protein (termed SASP-1) containing a single spore protease cleavage site as well as other sequences conserved in Bacillus megaterium SASPs A, C, C-1, C-2, and C-3. The B. subtilis SASP-1 genes's coding sequence is preceded by a potential Bacillus ribosome-binding site, and is followed by a sequence that could form a stem-and-loop structure characteristic of transcription termination sites. Upstream from the coding sequence there are no obvious homologies with other B. subtilis sporulation genes, but similarities with B. megaterium SASP genes are evident. SASP-1 mRNA (290 bases long) is absent from vegetative cells, but appears midway in sporulation and then disappears. The cloned SASP-1 gene hybridizes to three bands other than the SASP-1 gene itself in EcoRI or HindIII digests of B. subtilis DNA. Presumably these other bands represent SASP genes related to the SASP-1 gene, and we have been able to detect at least three such proteins in B. subtilis spores. PMID- 2981808 TI - Delta endotoxin of Bacillus thuringiensis subsp. israelensis. AB - From Bacillus thuringiensis subsp. israelensis, a proteinase-resistant protein was purified which exhibited toxicity to larval mosquitoes and cultured mosquito cells, lysed erythrocytes, and was lethal to mice. To extract the protein, a sporulating culture of B. thuringiensis subsp. israelensis was treated with alkali, neutralized, and incubated with trypsin and proteinase K. It was then purified by gel filtration and DEAE column chromatography. Up to 240 micrograms of toxic protein was purified from 1 g (wet weight) of culture pellet. Two closely related forms of toxic protein were obtained: the 25a and 25b proteins. The two forms comigrated near 25,000 daltons in a sodium dodecyl sulfate polyacrylamide gel, were serologically related, and showed similar partial protease digestion profiles, but were distinguishable by DEAE chromatography and nondenaturing polyacrylamide gel electrophoresis. Protein sequencing data indicated the 25b protein lacked the two amino acids at the amino terminus of the 25a protein. A Western blot enzyme-linked immunosorbent assay of alkali solubilized proteins that were not treated with proteases suggested the toxic 25a and 25b proteins were proteolytically derived from a larger molecule of about 28,000 daltons. Alkali-solubilized proteins from an acrystalliferous strain of B. thuringiensis subsp. israelensis and from B. thuringiensis subsp. kurstaki failed to cross-react with antibodies to the 25a protein. PMID- 2981807 TI - Isolation and characterization of outer membrane permeability mutants in Escherichia coli K-12. AB - Escherichia coli normally requires the lamB gene for the uptake of maltodextrins. We have identified and characterized three independent mutations that allow E. coli to grow on maltodextrin in the absence of a functional lamB gene by allowing maltodextrins with a molecular weight greater than 1,000 to cross the outer membrane barrier. Two of the mutations map to the structural gene for the outer membrane porin OmpF, and the remaining mutation maps to the structural gene for the second major outer membrane porin, OmpC. These mutations increase the permeability of the outer membrane to small hydrophilic substances, antibiotics, and detergents. These mutations alter the electrophoretic mobility of the respective porin proteins. PMID- 2981809 TI - Regulation of expression of the crp gene of Escherichia coli K-12: in vivo study. AB - Expression of the crp gene was studied in vivo by use of a crp-lacZ gene fusion first constructed on a plasmid and then transferred onto the chromosome. Our in vivo data confirm the in vitro findings that crp is negatively autoregulated via the cyclic AMP-catabolite gene activator protein complex. We present evidence that gene crp is repressed by glucose. PMID- 2981810 TI - hisT is part of a multigene operon in Escherichia coli K-12. AB - The Escherichia coli K-12 hisT gene has been cloned, and its organization and expression have been analyzed on multicopy plasmids. The hisT gene, which encodes tRNA pseudouridine synthase I (PSUI), was isolated on a Clarke-Carbon plasmid known to contain the purF gene. The presence of the hisT gene on this plasmid was suggested by its ability to restore both production of PSUI enzymatic activity and suppression of amber mutations in a hisT mutant strain. A 2.3-kilobase HindIII-ClaI restriction fragment containing the hisT gene was subcloned into plasmid pBR322, and the resulting plasmid (designated psi 300) was mapped with restriction enzymes. Complementation analysis with different kinds of hisT mutations and tRNA structural analysis confirmed that plasmid psi 300 contained the hisT structural gene. Enzyme assays showed that plasmid psi 300 overproduced PSUI activity by ca. 20-fold compared with the wild-type level. Subclones containing restriction fragments from plasmid psi 300 inserted downstream from the lac promoter established that the hisT gene is oriented from the HindIII site toward the ClaI site. Other subclones and derivatives of plasmid psi 300 containing insertion or deletion mutations were constructed and assayed for production of PSUI activity and production of proteins in minicells. These experiments showed that: (i) the proximal 1.3-kilobase HindIII-BssHII restriction fragment contains a promoter for the hisT gene and encodes a 45,000-dalton polypeptide that is not PSUI; (ii) the distal 1.0-kilobase BssHII-ClaI restriction fragment encodes the 31,000-dalton PSUI polypeptide; (iii) the 45,000 dalton polypeptide is synthesized in an approximately eightfold excess compared with PSUI; and (iv) synthesis of the two polypeptides is coupled, suggesting that the two genes are part of an operon. Insertion of mini-Mu d1 (lac Km) phage into plasmid psi 300 confirmed that the hisT gene is the downstream gene in the operon. PMID- 2981811 TI - Cyclic AMP receptor protein from yeast mitochondria: submitochondrial localization and preliminary characterization. AB - We have identified and characterized a cyclic AMP receptor protein in mitochondria of the yeast Saccharomyces cerevisiae. The binding is specific for cyclic nucleotides, particularly for cyclic AMP which is bound with high affinity (Kd of 10(-9) M) at 1 to 5 pmol/mg of mitochondrial protein. The mitochondrial cyclic AMP receptor is synthesized on cytoplasmic ribosomes and has an apparent molecular weight of 45,000 as determined by photoaffinity labeling. It is localized in the inner mitochondrial membrane and faces the intermembrane space. Cross-contamination of mitochondrial inner membranes by plasma membranes or soluble cytoplasmic proteins is excluded. PMID- 2981813 TI - Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetic acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134(pJP4). AB - Plasmid pJP4 permits its host bacterium, strain JMP134, to degrade and utilize as sole sources of carbon and energy 3-chlorobenzoate and 2,4-dichlorophenoxyacetic acid (R. H. Don and J. M. Pemberton, J. Bacteriol. 145:681-686, 1981). Mutagenesis of pJP4 by transposons Tn5 and Tn1771 enabled localization of five genes for enzymes involved in these catabolic pathways. Four of the genes, tfdB, tfdC, tfdD, and tfdE, encoded 2,4-dichlorophenol hydroxylase, dichlorocatechol 1,2-dioxygenase, chloromuconate cycloisomerase, and chlorodienelactone hydrolase, respectively. No function has been assigned to the fifth gene, tfdF, although it may encode a trans-chlorodiene-lactone isomerase. Inactivation of genes tfdC, tfdD, and tfdE, which encode the transformation of dichlorocatechol to chloromaleylacetic acid, prevented host strain JMP134 from degrading both 3 chlorobenzoate and 2,4-dichlorophenoxyacetic acid, which indicates that the pathways for these two substrates utilize common enzymes for the dissimilation of chlorocatechols. Studies with cloned catabolic genes from pJP4 indicated that whereas all essential steps in the degradation of 2,4-dichlorophenoxyacetic acid are plasmid encoded, the conversion of 3-chlorobenzoate to chlorocatechol is specified by chromosomal genes. PMID- 2981812 TI - Comparison of transformation mechanisms of Haemophilus parainfluenzae and Haemophilus influenzae. AB - Transformation pathways in two closely related bacterial species, Haemophilus parainfluenzae and Haemophilus influenzae, were studied. Both organisms rapidly take up transforming DNA within minutes into specialized membranous structures on the cell surface (transformasomes). DNA within transformasomes is in a protected state, inaccessible to external DNase or internal restriction and modification enzymes. However, the subsequent processing of donor DNA differs in these two organisms. In H. influenzae, linear DNA immediately undergoes degradation from one end at a constant rate, leaving a lower-molecular-weight intermediate in the transformasome. The end undergoing degradation is searching for homologous regions of the chromosome. Once pairing is initiated, the remaining lower molecular-weight DNA exits from the transformasome, and a single strand undergoes efficient integration. In contrast, in H. parainfluenzae little degradation of donor DNA is observed, with the majority remaining intact within the transformasomes after 1 h. Thus, whereas only 10% of donor DNA molecules leave the protected state after 1 h, portions of each molecule appear to become quantitatively integrated. PMID- 2981814 TI - Genetic analysis of Escherichia coli oligopeptide transport mutants. AB - The composition of the outer membrane channels formed by the OmpF and OmpC porins is important in peptide permeation, and elimination of these proteins from the Escherichia coli outer membrane results in a cell in which the primary means for peptide permeation through this cell structure has been lost. E. coli peptide transport mutants which harbor defects in genes other than the ompF/ompC genes have been isolated on the basis of their resistance to toxic tripeptides. The genetic defects carried by these oligopeptide permease-negative (Opp-) strains were found to map in two distinct chromosomal locations. One opp locus was trp linked and mapped to the interval between att phi 80 and galU. Complementation studies with F'123 opp derivatives indicated that this peptide transport locus resembles that characterized in Salmonella typhimurium as a tetracistronic operon (B. G. Hogarth and C. F. Higgins, J. Bacteriol. 153:1548-1551, 1983). The second opp locus, which we have designated oppE, was mapped to the interval between dnaC and hsd at 98.5 min on the E. coli chromosome. The differences in peptide utilization, sensitivity and resistance to toxic peptides, and the L-[U 14C]alanyl-L-alanyl-L-alanine transport properties observed with these Opp-E. coli strains demonstrated that the transport systems encoded by the trp-linked opp genes and by the oppE gene(s) have different substrate preferences. Mutants harboring defects in both peptide transport loci defined in this study would not grow on nutritional peptides except for tri-L-methionine, were totally resistant to toxic peptides, and would not actively transport L-[U-14C]alanyl-L-alanyl-L alanine. PMID- 2981815 TI - Purification and characterization of two alpha-galactosidases associated with catabolism of guar gum and other alpha-galactosides by Bacteroides ovatus. AB - When Bacteroides ovatus is grown on guar gum, a galactomannan, it produces alpha galactosidase I which is different from alpha-galactosidase II which it produces when grown on galactose, melibiose, raffinose, or stachyose. We have purified both of these enzymes to apparent homogeneity. Both enzymes appear to be trimers and have similar pH optima (5.9 to 6.4 for alpha-galactosidase I, 6.3 to 6.5 for alpha-galactosidase II). However, alpha-galactosidase I has a pI of 5.6 and a monomeric molecular weight of 85,000, whereas alpha-galactosidase II has a pI of 6.9 and a monomeric molecular weight of 80,500. alpha-Galactosidase I has a lower affinity for melibiose, raffinose, and stachyose (Km values of 20.8, 98.1, and 8.5 mM, respectively) than does alpha-galactosidase II (Km values of 2.3, 5.9, and 0.3 mM, respectively). Neither enzyme was able to remove galactose residues from intact guar gum, but both were capable of removing galactose residues from guar gum which had been degraded into large fragments by mannanase. The increase in specific activity of alpha-galactosidase which was associated with growth on guar gum was due to an increase in the specific activity of enzyme I. Low, constitutive levels of enzyme II also were produced. By contrast, enzyme II was the only alpha-galactosidase that was detectable in bacteria which had been grown on galactose, melibiose, raffinose, or stachyose. PMID- 2981816 TI - Characterization of xylitol-utilizing mutants of Erwinia uredovora. AB - Of the four pentitols ribitol, xylitol, D-arabitol, and L-arabitol, Erwinia uredovora was able to utilize only D-arabitol as a carbon and energy source. Although attempts to isolate ribitol- or L-arabitol-utilizing mutants were unsuccessful, mutants able to grow on xylitol were isolated at a frequency of 9 X 10(-8). Xylitol-positive mutants constitutively synthesized both a novel NAD dependent xylitol-4-dehydrogenase, which oxidized xylitol to L-xylulose, and an L xylulokinase. The xylitol dehydrogenase had a Km for xylitol of 48 mM and showed best activity with xylitol and D-threitol as substrates. However, D-threitol was not a growth substrate for E. uredovora, and its presence did not induce either dehydrogenase or kinase activity. Attempts to determine the origin of the xylitol catabolic enzymes were unsuccessful; neither enzyme was induced on any growth substrate or in the presence of any polyol tested. Analysis of xylitol-negative mutants isolated after Tn5 mutagenesis suggested that the xylitol dehydrogenase and the L-xylulokinase structural genes were components of two separate operons but were under common regulatory control. PMID- 2981817 TI - Identification of the transcriptional suppressor sof-1 as an alteration in the spo0A protein. AB - The mutation sof-1 suppresses the sporulation defect of mutations in either the spo0B, spo0E, or spo0F stage 0 sporulation genes. Through the use of integrative plasmids carrying the portion of the chromosome including the spo0A locus and flanking regions, the sof-1 mutation was localized near the spo0A locus. A plasmid carrying a fragment of DNA with sof genetic activity was constructed. Nucleic acid sequence analysis of this fragment revealed a single base change that resulted in a substitution of lysine for asparagine in the 12th codon of the spo0A gene. The results indicate that certain missense mutations in the spo0A gene bypass the necessity for the spo0B, spo0E, and spo0F gene products in sporulation. Several models for the interaction of these gene products may be imagined. PMID- 2981818 TI - Uptake of phenol by Trichosporon cutaneum. AB - The soil yeast Trichosporon cutaneum, which is distinguished by having a strictly oxidative metabolism, can be induced to utilize phenol as a sole carbon source. The present paper shows that such phenol-induced cells contain a specific, energy dependent uptake system for phenol. Phenol uptake is not directly linked to its o hydroxylation inside the cell, the first step of phenol metabolism. The Km for uptake is 235 +/- 30 microM, that for hydroxylation only 4.5 +/- 0.5 microM. Further, the phenol analog 2,6-dimethylphenol, which can not be hydroxylated, competes with phenol for the uptake system. The pH dependence of uptake indicates that phenolate is an essential form during the uptake process. The energy requirement for phenol uptake is indicated by effects of various inhibitors of energy generation, including proton-conducting uncouplers. Direct monitoring of proton movements in a pH-stat during phenol uptake indicates a phenol-proton symport. One proton is cotransported with every phenol molecule. Phenol competes with the uptake of sucrose and glycerol by cells grown on these substrates. Under such conditions the uptake of phenol seems to proceed through a different system, with lower affinity for phenol than in phenol-grown cells. PMID- 2981820 TI - Proton circulation in Vibrio costicola. AB - The importance of proton movements was assessed in the moderate halophile Vibrio costicola. When anaerobic cells in acidic buffer (pH 6.5) were given an O2 pulse, protons were extruded regardless of the presence of Na+. At pH 8.5, however, V. costicola produced an acidic response to an O2 pulse in the absence of Na+ and an alkaline response when Na+ was present. An Na+/H+ antiport activity was confirmed at pH 8.5. All of these effects were prevented by protonophores or butanol treatment. Growth in complex medium at pH 8.5 was prevented by a high concentration (50 microM) of carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) or a low concentration (5 microM) of another protonophore, 3,3',4',5 tetrachlorosalicylanilide (TCS). The relative ineffectiveness of the former protonophore was caused by the proteose peptone and tryptone ingredients of the complex medium, since 5 microM completely prevented growth in their absence. The results are explained by a primary respiratory-linked proton efflux coupled to a secondary Na+/H+ antiport operating at alkaline pH. Evidence was seen for a role of Na+ in stimulating proton influx at alkaline pH, presumably via the pH homeostasis mechanism. PMID- 2981819 TI - Regulation of a cya-lac fusion by cyclic AMP in Salmonella typhimurium. AB - cya-lac and crp-lac operon fusions were isolated in Salmonella typhimurium by using the phage Mu d1(lac cts Apr). Both transduction and reversion analyses have indicated that lac expression is controlled by the appropriate promoter, e.g., either crpp or cyap. By using chromosomal mobilization techniques, we found that cya had a clockwise direction of transcription on the standard S. typhimurium map. The cya-lac fusions could be complemented by Escherichia coli F'133, which covers cya, with a resultant 17 to 38% decrease in cya expression. Cyclic AMP was found to be able to repress the expression of the cya-lac fusion ninefold when present at 25 mM. This repression was not seen in crp backgrounds, and hence is mediated by the cAMP receptor protein. Repression of cya was also found upon growth on carbon sources known to elicit high cyclic AMP levels. PMID- 2981822 TI - Immunological investigation of the distribution of cytochromes related to the two terminal oxidases of Escherichia coli in other gram-negative bacteria. AB - Monospecific antibodies were raised against the two terminal oxidase complexes of the aerobic respiratory chain of Escherichia coli. These are the cytochrome d and cytochrome o complexes. The antibodies were used to check for the occurrence of cross-reactive antigens in membrane preparations from a variety of gram-negative bacteria by rocket immunoelectrophoresis and immunoblotting techniques. With these criteria, proteins closely related to the cytochrome d complex of E. coli appeared to be widely distributed. Among the strains containing cytochrome d related material were Serratia marcescens, Photobacterium phosphoreum, Salmonella typhimurium, Klebsiella pneumoniae, and Azotobacter vinelandii. The data suggest that the d-type terminal oxidase in many of these strains is associated in a complex with b-type and a1-type cytochromes, as has been found to be the case in E. coli. K. pneumoniae and S. typhimurium were also shown to have material cross reactive to the E. coli cytochrome o complex. PMID- 2981823 TI - Maltose-binding protein does not modulate the activity of maltoporin as a general porin in Escherichia coli. AB - Maltoporin (lambda receptor) is part of the maltose transport system in Escherichia coli and is necessary for the facilitated diffusion of maltose and maltodextrins across the outer membrane. Maltoporin also allows the diffusion of nonmaltodextrin substrates, albeit with less efficiency. The preference of maltoporin for maltodextrins in vivo is thought to be the result of an interaction of maltoporin with the maltose-binding protein, the malE gene product. In a recent report Heuzenroeder and Reeves (J. Bacteriol. 144:431-435, 1980) suggested that this interaction establishes a gating mechanism which inhibits the diffusion of nonmaltodextrin substrates, such as lactose. To reinvestigate this important conclusion, we constructed ompR malTc strains carrying either the malE+ gene, the nonpolar malE444 deletion, or the malE254 allele, which specifies an interaction-deficient maltose-binding protein. Lactose uptake was measured at different concentrations below the Km of this transport system and under conditions where transport was limited by the diffusion through maltoporin. We found no difference in the kinetics of lactose uptake irrespective of the malE allele. We conclude that the maltose-binding protein does not modulate the activity of maltoporin as a general outer membrane porin. PMID- 2981821 TI - Involvement of DNA superhelicity in minichromosome maintenance in Escherichia coli. AB - Evidence is presented that Escherichia coli minichromosomes are harbored at superhelical densities which are lower than those measured for other E. coli plasmids but are comparable to that of the chromosome. When introduced into gyrB decreased-supercoiling mutants, minichromosomes were much more unstable than in strains with normal or increased supercoiling properties; in fact, certain minichromosome derivatives could not be introduced into top gyrB decreased supercoiling mutants. These observations were unique to minichromosomes, since the maintenance of plasmids which did not replicate from oriC was not altered in these mutants. Analyses of minichromosomes of identical sizes but with different restriction fragment orientations suggested that supercoiling-dependent alterations in promoter-terminator functions, as well as direct effects of supercoiling on replication, may play a role in the observed minichromosome instability. PMID- 2981824 TI - Membrane proteins correlated with expression of the polysialic acid capsule in Escherichia coli K1. AB - Growth of Escherichia coli K1 strains at 15 degrees C results in a defect in the synthesis or assembly of the K1 polysialic acid capsule. Synthesis is reactivated in cells grown at 15 degrees C after upshift to 37 degrees C, and activation requires protein synthesis (Whitfield et al., J. Bacteriol. 159:321-328, 1984). Using this temperature-induced defect, we determined the molecular weights and locations of membrane proteins correlated with the expression of K1 (polysialosyl) capsular antigen. Pulse-labeling experiments demonstrated the presence of 11 proteins whose synthesis was correlated with capsule appearance at the cell surface. Using the differential solubility of inner and outer membranes in the detergent Sarkosyl, we localized five of the proteins in the outer membrane and four in the inner membrane. The subcellular location of two of the proteins was not determined. Five proteins appeared in the membrane simultaneously with the initial expression of the K1 capsule at the cell surface. One of these proteins, a 40,000-dalton protein localized in the outer membrane, was identified as porin protein K, which previously has been shown to be present in the outer membrane of encapsulated E. coli. The possible role of these proteins in the synthesis of the polysialosyl capsule is discussed. PMID- 2981825 TI - Enzymatic detection of phospholipid biosynthetic enzymes after electroblotting. AB - The membrane-associated enzymes phosphatidylinositol synthase (CDPdiacylglycerol:myo-inositol 3-phosphatidyltransferase; EC 2.7.8.11) and phosphatidylserine synthase (CDPdiacylglycerol:L-serine O phosphatidyltransferase; EC 2.7.8.8) from Saccharomyces cerevisiae were detected enzymatically after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting. Enzyme activities were measured on nitrocellulose blots by using pure enzyme preparations as well as Triton X-100-solubilized membranes. Phosphatidylinositol synthase activity migrated to Mr 34,000, and phosphatidylserine synthase activity migrated to Mr 23,000. PMID- 2981826 TI - Salmonella typhimurium contains an anion-selective outer membrane porin induced by phosphate starvation. AB - A mutant of Salmonella typhimurium was selected that is constitutive for the pho regulon. It exhibited constitutive glycerol-3-phosphate transport activity and synthesized a new outer membrane porin. Upon measurement of porin activity in black lipid films, it exhibited anion selectivity. It therefore appears analogous to the Escherichia coli PhoE porin. PMID- 2981827 TI - Chlorpromazine vs. meperidine in the treatment of phencyclidine psychosis. AB - Patients with phencyclidine psychosis were treated with two 50 mg injections of either chlorpromazine (N = 10) or meperidine (N = 10). The chlorpromazine-treated group responded more rapidly, but the meperidine-treated group had greater overall improvement. It is suggested that meperidine may have a role in the emergency treatment of phencyclidine psychosis. These findings may provide support for the hypothesis that dopaminergic psychosis is mediated by opioids. PMID- 2981828 TI - Hypothalamic factor inhibits the (Na,K)ATPase from the extracellular surface. Mechanism of inhibition. AB - We have characterized the effect of a stable small molecule isolated from bovine hypothalamus (Haupert, G. T., and Sancho, J. M. (1979) Proc. Natl. Acad. Sci. 76, 4658-4660) on mammalian (Na,K)ATPase. This hypothalamus-derived inhibitory factor, HIF, has been shown to inhibit ATPase activity of purified dog kidney enzyme reversibly with high affinity (Haupert, G. T., Carilli, C. T., and Cantley, L. C. (1984) Am. J. Physiol. 247, F919-F924). In this report it is shown that HIF inhibits the ouabain sensitive component of 86Rb+ uptake into human red blood cells. HIF also inhibited (Na,K)ATPase activity of unsealed red cell membranes but not that of sealed inside-out vesicles, indicating that HIF is impermeant to red cell membranes and inhibits the (Na,K)ATPase from the extracellular side. In unsealed human red cell membranes, concentrations of HIF which caused 70% inhibition of the (Na,K)ATPase did not inhibit ATP hydrolysis by plasma membrane (Ca2+)ATPase or (Mg2+)ATPase. However, at a similar concentration, HIF was shown to inhibit rabbit muscle sarcoplasmic reticulum (Ca2+)ATPase. HIF also inhibited p-nitrophenylphosphatase activity of unmodified or fluorescein-5'-iso-thiocyanate labeled dog kidney (Na,K)ATPase. As judged by fluorescein fluorescence of the modified enzyme, HIF stabilized the low fluorescent "E2" conformation of the enzyme similar to that stabilized by ouabain. However, unlike ouabain, HIF blocked covalent phosphorylation of dog kidney (Na,K)ATPase by inorganic phosphate. These studies show that HIF is an inhibitor of (Na,K)ATPase which acts from the extracellular side of the membrane by a mechanism similar to but not identical to that of cardiac glycosides. PMID- 2981829 TI - Phosphoinositide interconversion in thrombin-stimulated human platelets. AB - Stimulation of platelets and other secretory cells by agonists results in the degradation of phosphoinositides by phospholipase C. Kinetic studies suggest that hydrolysis of phosphatidylinositol 4,5-diphosphate (PI-4,5-P2) is an initial event in this process. Platelets contain much larger amounts of phosphatidylinositol (PI) than PI-4,5-P2, and approximately 50% of total phosphoinositides are degraded upon stimulation. We have investigated whether degradation of PI occurs by direct phospholipase C hydrolysis or by phosphorylation to PI-4,5-P2 followed by phospholipase C action on the latter compound. When platelets are incubated for 3 min with 32Pi prior to stimulation, the phosphoinositides are labeled to different specific activities. Under these nonequilibrium conditions, the time course of change in specific activity reflects turnover. The rise in specific activity of phosphatidylinositol 4 phosphate (PI-4-P) is similar in stimulated and unstimulated cells, indicating that there is little increase in the conversion of PI to PI-4-P during thrombin stimulation. In addition, the specific activity of the 4-phosphate in PI-4-P during thrombin stimulation is less than both the 5-phosphate of PI-4,5-P2 and the phosphate group of phosphatidic acid, indicating that the 4-phosphate moiety is not labeled to equilibrium with ATP. This finding is inconsistent with a rapid flux of PI via PI-4-P to PI-4,5-P2 during thrombin stimulation, in which case the 4-phosphate would be at maximum specific activity. We, therefore, conclude that the bulk of PI breakdown that occurs in thrombin-stimulated platelets occurs via direct phospholipase C hydrolysis of PI. PMID- 2981830 TI - Degradation of monomeric and fibrillar type III collagens by human skin collagenase. Kinetic constants using different animal substrates. AB - Human skin collagenase activity was examined against type III collagens, in both soluble and fibrillar form, from different animal species. In either form, human, dog, and cat type III were degraded 10- to 30-fold faster than was that from guinea pig and nearly 100-fold more readily than chick type III. These differences in susceptibility were mirrored by essentially identical differences in the rate of trypsin cleavage of the same substrates. Human, dog, and cat type III were cleaved most rapidly by trypsin, guinea pig III more slowly, and chick III was completely resistant to the serine protease. Arrhenius plots, relating enzyme activity to temperature, revealed differences in the various type III substrates consistent with their collagenase and trypsin susceptibilities. Human, dog, and cat type III collagens yielded nonlinear plots, with accompanying activation energies which decreased at temperatures above 26 degrees C; guinea pig type III displayed a plot which deviated only slightly from linearity while the plot for chick type III was completely linear. These data strongly suggest that type III collagens display substantial variability in the stability of the helix at or near the collagenase cleavage site. The susceptibility of these type III substrates as reconstituted fibrils was also examined. The relative rates of degradation of these substrates by collagenase, and by trypsin, were the same as those observed in solution. The absolute rates of degradation of collagen in fibrillar form, however, were massively lower than predicted by extrapolation from solution values. This reduction in rate is even greater for type III than for type I collagens. Thus, whereas in solution type III substrates are cleaved much faster than type I collagens, in fibrillar form these differences are less than 2-fold. These data, together with values for activation energies and deuterium isotope effects on type III fibrillar substrates, reinforce the concept that helical integrity near the collagenase cleavage site is a major specifier of the rate of collagenase activity. Furthermore, the data suggest that the exclusion of water accompanying the tight packing of monomers into fibrils presents a major energy barrier to collagenase activity, which is particularly large for type III collagen. PMID- 2981831 TI - Identification of two proteins (actin-binding protein and P235) that are hydrolyzed by endogenous Ca2+-dependent protease during platelet aggregation. AB - Our previous research has shown that the Ca2+-dependent protease within platelets is activated when platelets aggregate, resulting in the production of three polypeptides (Mr = 200,000, 100,000, and 91,000). We have now shown that these three polypeptides arise from the hydrolysis of actin-binding protein. An antibody against actin-binding protein raised in rabbits was shown to be specific for actin-binding protein on immunoblots of total platelet proteins. This antibody reacted with additional polypeptides of Mr = 200,000, 100,000, and 91,000 on immunoblots of the proteins of thrombin-activated platelets. Actin binding protein was purified from fresh, human platelet concentrate and hydrolyzed with platelet-derived Ca2+-dependent protease; hydrolysis resulted in the appearance of three polypeptides with molecular weights and isoelectric points identical to those of the three polypeptides generated within intact, aggregating platelets. Production of these polypeptides was inhibited by leupeptin and by the chelation of Ca2+. Hydrolysis of actin-binding protein was observed at micromolar Ca2+ concentrations, demonstrating that the level of Ca2+ in aggregated platelets is sufficient to account for the hydrolysis of actin binding protein by the Ca2+-dependent protease. P235 was also purified and tested for its susceptibility to the protease. It was hydrolyzed by the Ca2+-dependent protease, and two polypeptides (Mr = 200,000 and 46,000) were produced. Antibodies against P235 raised in rabbits reacted only with P235 on immunoblots of total platelet proteins. These antibodies also reacted with polypeptides of Mr = 200,000 and 46,000 on immunoblots of thrombin-activated platelets. These data show that both actin-binding protein and P235 are cleaved during thrombin-induced platelet aggregation and suggest that the activation of the Ca2+-dependent protease may permit reorganization of the platelet cytoskeleton in aggregating platelets. PMID- 2981832 TI - Expression of an ouabain resistance gene in transfected cells. Ouabain treatment induces a K+-transport system. AB - We have investigated the expression of a cloned mouse gene which confers ouabain resistance to African green monkey kidney (CV1) cells. CV1 cells carrying the transfected ouabain resistance (ouaR) gene express an ouabain-inducible K+ transport system. This K+-transport system is not a normal (Na,K)-ATPase since plasma membranes prepared from the transfected cells have significantly reduced Na+-stimulated ATPase activity. RNA sequences homologous to the transfected gene are observed in abundance only following exposure of transfectants to ouabain. The small size of the message induced (1.2 kilobases) also argues that the gene does not code for the alpha-subunit of the (Na,K)-ATPase. Ouabain-treated transfected cells maintain an internal [K+] of 113 mM; a level close to the 139 mM of control cells. However, ouabain-treated transfectants exhibit an internal [Na+] of 61 mM, which is 3-6 times the level in untreated cells (11-21 mM). These results suggest: ouabain resistance can be conferred by a gene which codes for an ouabain-inducible K+-transport system; induction of this transport system by ouabain is due to increased levels of mRNA coded for by the ouabain resistance gene; and the ouabain resistance gene does not encode for the alpha-subunit of the (Na,K)-ATPase. PMID- 2981833 TI - Simian virus 40 maturation in cells harboring mutants deleted in the agnogene. AB - The predominant leader region of the late 16 S mRNAs of simian virus 40 encodes a histone-like, 61-amino acid, DNA-binding protein called the agnoprotein or LP1. To test the hypothesis that this protein facilitates assembly of viral minichromosomes into virions, we have studied the synthesis of virions in cells infected with mutants deleted in this region of the SV40 genome. We found that 220 S mature virions, indistinguishable from those of wild type, were produced in cells infected with these mutants. As in wild-type-infected cells, no assembly intermediates other than 75 S chromatin were observed. However, data obtained from both steady-state and pulse-chase labeling experiments indicated that cells infected with agnogene deletion mutants produced virions more slowly than cells infected with wild-type virus. Taken together with data showing that similar levels of virion proteins were present in the wild-type- and mutant-infected cells, these findings strongly suggest that LP1 plays a role in expediting virion assembly. PMID- 2981834 TI - Amiloride inhibits phorbol ester-stimulated Na+/H+ exchange and protein kinase C. An amiloride analog selectively inhibits Na+/H+ exchange. AB - The human leukemic cell line, HL-60, differentiates in response to tumor promoting phorbol esters. Recently, we have reported that one of the first events evoked by phorbol esters in HL-60 cells is the stimulation of Na+-dependent H+ efflux. In efforts to determine whether stimulation of Na+/H+ exchange by phorbol esters is coupled to induction of cellular differentiation, we found that 1) amiloride, a frequently used inhibitor of Na+/H+ exchange, rapidly inhibits phorbol ester-stimulated protein phosphorylation in vivo and protein kinase C mediated phosphorylation in vitro, both with potency similar to that with which amiloride inhibits Na+/H+ exchange; 2) an amiloride analog, dimethylamiloride, is a far more potent inhibitor of Na+/H+ exchange than is amiloride, while being no more potent than amiloride in inhibiting phorbol ester/protein kinase C-mediated phosphorylation; and 3) at concentrations sufficient to completely inhibit Na+/H+ exchange, amiloride blocked phorbol ester-induced adhesion of HL-60 cells (adhesion being a property indicative of the differentiated state), but dimethylamiloride (as well as ethylisopropylamiloride, another very potent amiloride analog) did not. Thus, dimethylamiloride represents a potential tool for distinguishing protein kinase C-coupled from Na+/H+ exchange-coupled events in phorbol ester-stimulated cells. PMID- 2981835 TI - An evolutionary switch in tissue-specific gene expression. Abundant expression of alpha 1-antitrypsin in the kidney of a wild mouse species. AB - alpha 1-Antitrypsin (AT), one of the major proteinase inhibitors in mammalian serum, is generally considered to be synthesized exclusively in the liver. We have found that a wild-derived Mus species, Mus caroli, expresses AT mRNA in kidney at levels approaching that in liver; no other mouse, inbred or wild derived, exhibits this striking property. Liver and kidney mRNAs from M. caroli encode very similar AT polypeptides that are distinct from that encoded by Mus musculus liver mRNA. In vivo, liver AT is secreted into the bloodstream, while kidney AT, which is processed differently from the liver protein, is excreted into the urine. Analysis of RNA from a hybrid between M. musculus and M. caroli indicates that a cis-acting genetic element may be responsible for the difference in AT expression. Restriction enzyme digestion patterns of AT genomic sequences in M. caroli DNA are considerably different from those in M. musculus; in addition, these sequences are undermethylated in liver DNA from M. musculus and in liver and kidney DNA from M. caroli, reflecting the respective patterns of expression. Further studies of the altered tissue specificity of AT expression that is apparent in these two related species should lead to new insights into the nature and evolution of genetic determinants of tissue-specific phenotypes. PMID- 2981836 TI - Co-existence of two different types of soluble histone complexes in nuclei of Xenopus laevis oocytes. AB - The nuclear pool of soluble histones in Xenopus laevis oocytes is organized into two major types of acidic histone complexes separable by sucrose density gradient centrifugation. One type of complex sediments at 5 S (Mr approximately 120,000), is isoelectric at pH 4.6, and contains histones H3 and/or H4 tightly bound to one polypeptide of a pair of very acidic polypeptides, designated N1 and N2 (Kleinschmidt, J. A., and Franke, W. W. (1982) Cell 29, 799-809). This complex can be selectively immunoprecipitated by guinea pig antibodies against purified protein N1/N2. In contrast, a larger complex of 7 S contains four histones and nucleoplasmin (the purified protein exists as a pentamer of a polypeptide of Mr approximately 30,000), is isoelectric over the pH range of 5-7, and can be immunoprecipitated by nucleoplasmin antibodies. Its relative molecular weight of 130,000-170,000, as determined by gel filtration, sucrose density gradient centrifugation, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the cross-linked complexes, excludes the association of a histone octamer with nucleoplasmin. In addition to histones H2A and H2B, two histones (designated H3 and H4) which are similar in their electrophoretic mobilities to histones H3 and H4 but have lower isoelectric pH values are enriched in immuno-precipitates obtained with nucleoplasmin antibodies. Cross-linking of complexes present in intact nuclei, using 1% formaldehyde at near-physiological ionic strength and pH, indicates the coexistence of these two soluble histone complexes in the living cell. In chromatin assembly experiments using SV 40 DNA, both histone fractions are able to transfer histones to DNA, resulting in an increase of DNA superhelicity and the formation of beaded nucleoprotein complexes of nucleosome like morphology. The common principle governing both types of complexes, i.e. the association of one or two histone molecules with a karyophilic large acidic histone-binding protein is emphasized. We discuss the possible role of these complexes in storing histones utilized in chromatin assembly during early amphibian embryogenesis as well as the possible existence of similar complexes, albeit at lower concentrations, in somatic cells. PMID- 2981837 TI - Identification of two molecular forms of (Na+,K+)-ATPase in rat adipocytes. Relation to insulin stimulation of the enzyme. AB - Two molecular forms of the (Na+,K+)-ATPase catalytic subunit have been identified in rat adipocyte plasma membranes using immunological techniques. The similarity between these two forms and those in brain (Sweadner, K. J. (1979) J. Biol. Chem. 254, 6060-6067) led us to use the same nomenclature: alpha and alpha(+). The K0.5 values of each form for ouabain (determined by inhibition of phosphorylation of the enzyme from [gamma-32P]ATP) were 3 X 10(-7)M for alpha(+) and 1 X 10(-5)M for alpha. These numbers correlate well with the K0.5 values for the two ouabain inhibitable components of 86Rb+/K+ pumping in intact cells (1 X 10(-7) M and 4 X 10(-5)M). Quantitation of the Na+ pumps in plasma membranes demonstrated a total of 11.5 +/- 0.2 pmol/mg of membrane protein, of which 8.5 +/- 0.3 pmol/mg, or 75%, was alpha(+). Insulin stimulation of 86Rb+/K+ uptake in rat adipocytes was abolished by ouabain at a concentration sufficient to inhibit only alpha(+)(2-5 X 10(-6)M). Immunological techniques and ouabain inhibition of catalytic labeling of the enzyme from [gamma-32P]ATP demonstrated that alpha(+) was present in skeletal muscle membranes as well as in adipocyte membranes, but was absent from liver membranes. Since insulin stimulates increased Na+ pump activity in adipose and muscle tissue but not in liver, there is a correlation between hormonal regulation of (Na+,K+)-ATPase and the presence of alpha(+). We propose that alpha(+) is the hormonally-sensitive version of the enzyme. PMID- 2981838 TI - Covalent cross-linking of vasoactive intestinal polypeptide to its receptors on intact human lymphoblasts. AB - 125I-labeled vasoactive intestinal polypeptide (125I-VIP) was covalently cross linked with its binding sites on intact cultured human lymphoblasts by each of three bifunctional reagents: disuccinimidyl suberate (DSS), ethylene glycol bis(succinimidyl succinate) (EGS), and N-succinimidyl 6-(4'-azido-2' nitrophenylamino) hexanoate (SANAH). A fourth cross-linking agent with a shorter chain length, N-hydroxysuccinimidyl 4-azidobenzoate (HSAB), was much less effective in cross-linking 125I-VIP to the site. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography demonstrated a band of Mr approximately equal to 50,000 +/- 3,000, regardless of which cross linker was used. The labeling of this band was specific in that it was prevented by 10(-6) M unlabeled VIP and was partially blocked by the homologous hormones secretin and glucagon. The relative potencies of these peptides in blocking the cross-linking of 125I-VIP to the Mr approximately equal to 50,000 band of the lymphoblasts (VIP greater than secretin greater than or equal to glucagon) were similar to those previously found for competitive inhibition of 125I-VIP binding to its putative high-affinity receptor on these cells. The covalent cross-linking required a bifunctional reagent; it was dependent on both the number of Molt cells and the concentration of 125I-VIP. The apparent molecular weight of the cross-linked species was unchanged by treatment with dithiothreitol. These observations suggest that the Mr = 50,000 species represents 125I-VIP cross linked to a specific plasma membrane receptor and that the receptor does not contain interchain disulfide bonds. PMID- 2981839 TI - Nucleolytic processing of a tRNAArg-tRNAAsp dimeric precursor by a homologous component from Saccharomyces cerevisiae. AB - A subcellular extract from Saccharomyces cerevisiae has been used to transcribe cloned yeast tRNA genes in vitro and to process the primary transcripts at the 5' and 3' termini. Chromatographic fractionation of the extract has separated the transcription components from two distinct nucleolytic activities: an endonuclease that cleaves the precursors to produce mature 5' termini; and a 3' 5' exonuclease. These fractions have been used to elaborate a processing pathway for the dimeric primary transcript of the yeast tRNAArg-tRNAAsp gene pair. Under optimal conditions in vitro this gene is expressed at a rate of 200 transcripts/gene/hour, initiating at position -10 with respect to the mature 5' terminus of tRNAArg and terminating near position +160. The primary transcripts are cleaved by an endonuclease to give tRNAAsp with a mature 5' terminus, and a pre-tRNAArg monomer with a 5' leader and 3' trailer sequences. A second endonuclease cleavage of pre-tRNAArg generates the mature 5' terminus of tRNAArg. The endonuclease cleavages are not ordered. Exonuclease activity(ies) remove the spacer sequences from the 5' mature tRNAArg, and trim the 3' trailer portion from tRNAAsp. Exonucleolytic removal of the 3' trailer does not require prior endonuclease action, but removal of the spacer sequences from pre-tRNAArg is incomplete without prior removal of the 5' leader sequences. PMID- 2981840 TI - A gene encoding rat cholecystokinin. Isolation, nucleotide sequence, and promoter activity. AB - The gene for rat cholecystokinin (CCK) was isolated from a rat genomic DNA library. The transcription unit spans 7 kilobases and is interrupted by two introns. The initiator methionine codon lies 2 bases into exon 2; therefore, exon 1 is a noncoding exon. The transcription initiation site was determined using avian myeloblastosis reverse transcriptase, a cDNA primer, and mRNA isolated from a rat medullary thyroid carcinoma. A "TATA"-like sequence precedes the transcription initiation site at position -34. The polyadenylation site for the gene was mapped by a nuclease protection assay using a cRNA generated by transcription of the exon 3 region of the CCK gene with SP6 bacteriophage RNA polymerase. The sequence AT-TAAA is found 22 bases 5' to the site determined to be the polyadenylation addition site. Two regions of simple repetitive DNA occur within the CCK lambda clone, one within intron 2 and the other 4 kilobases 3' to the gene. Sequence analysis of the repetitive element 3' distal to the gene revealed two copies of the sequence 5'-(AC)n-3', where n is 22 and 25. A 114-base pair sequence of predominantly repeating purine-pyrimidine nucleotides separates these two d(AC) repeats. Transcriptional control elements were investigated by fusing regions of the CCK gene to the structural gene encoding chloramphenicol acetyltransferase. Promoter activity was determined by transfecting COS-7 cells with plasmids containing the gene fusions, followed by determining chloramphenicol acetyltransferase activity in cellular extracts. The region necessary for expression of the CCK gene fusions in COS-7 cells is within 144 bases 5' to the initiation of transcription. PMID- 2981841 TI - Cryptic satellites rich in inverted repeats comprise 30% of the genome of a hermit crab. AB - One major very highly repeated (VHR) DNA (approximately 7 X 10(6) copies/genome; repeat unit = 156 base pairs (bp)), a family of three minor VHR DNAs (approximately 2.8 X 10(6) copies/genome; repeat units = 71-74 bp), and a number of trace components account for almost 30% of the genome of a hermit crab. The repeat units of the three minor variants are defined by identical 14-bp G + C rich inverted repeats that might form cruciforms. Two copies of the repeat unit (CCTA) of one of two patent satellites of this crab (Skinner, D. M., and Beattie, W. G. (1974) Biochemistry 13, 3922-3929; Skinner, D. M., Beattie, W. G., Blattner, F. R., Stark, B. P., and Dahlberg, J. E. (1974) Biochemistry 13, 3930 3937) occur at the center of one in seven of the G + C-rich inverted repeats; copies of the other patent satellite (Chambers, C. A., Schell, M. P., and Skinner, D. M. (1978) Cell 13, 97-110) are found in main component DNA. The sequences of both the major and minor VHR DNAs are characterized by short tracts of An and/or Tn (n = 4-7) residues whose presence would permit the formation of perfectly matched stems separated by loops of 8-16 bp. The An and/or Tn tracts are interspersed with segments of G + C-rich DNA and are arranged differently in the major and minor VHR DNAs. Although the repeat units of the major and the three minor VHR DNAs are arranged in tandem, the composition and sequence of their bases are such that they do not form distinct bands in CsCl gradients; they are cryptic satellites. PMID- 2981842 TI - Post-translational processing of the epidermal growth factor receptor. Glycosylation-dependent acquisition of ligand-binding capacity. AB - The post-translational processing of the epidermal growth factor receptor in human A431 epidermoid carcinoma cells has been investigated. By employing the affinity matrix epidermal growth factor Affi-Gel in conjunction with immunoprecipitation, it has been demonstrated that core oligosaccharide addition is essential for the acquisition of epidermal growth factor-binding activity. Furthermore, the initial 160-kDa translation product was observed to undergo a processing step by which ligand-binding activity was acquired with a half-time of approximately 30 min while exhibiting no apparent change in mobility on sodium dodecyl sulfate-polyacrylamide gels. This was shown not to involve the conversion of high-mannose chains to complex chains which have been capped with fucose and sialic acid. Possible explanations for this activation in terms of translocation of intermediates and/or formation of disulfide bonds are discussed. PMID- 2981843 TI - Multiexon deletion in an osteogenesis imperfecta variant with increased type III collagen mRNA. AB - Recently, the dermal fibroblasts (ATCC CRL 1262) of a lethal perinatal variant of osteogenesis imperfecta have been used for the first molecular characterization of a collagen gene defect (Chu, M. L., Williams, C. J., Pepe, G., Hirsch, J. L., Prockop, D. J., and Ramirez, F. (1983) Nature (Lond.) 304, 78-80). These studies revealed that the patient was heterozygous for an internal deletion of approximately 500 base pairs in the pro-alpha 1(I) collagen gene, consistent with previous investigations indicating that CRL 1262 fibroblasts equally synthesized a normal and a shortened pro-alpha 1(I) chain (Barsh, G. S., and Byers, P. H. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 5142-5146). Cloning and analysis of the affected allele of CRL 1262 has now indicated that the deletion is contained between two introns of the pro-alpha 1(I) gene and results in the elimination of three exons of the triple helical domain. Furthermore, the termini of the rearrangement are located within two short inverted repeats suggesting that the self-complementary nature of these DNA elements may have favored the formation of a DNA secondary structure intermediate which, in turn, served as substrate for the deletion. Evidence are also presented for an elevated Type III collagen mRNA content in the patient fibroblasts. PMID- 2981844 TI - Isoforms of apolipoprotein A-II in human plasma and thoracic duct lymph. Identification of proapolipoprotein A-II and sialic acid-containing isoforms. AB - Proapolipoprotein (apo-) A-II and several isoforms of apo-A-II including sialylated isoforms were identified in human plasma and thoracic duct lymph. Proapo-A-II secreted by HepG2 cells was identified by a combination of immunoblots and [14C]arginine incorporation. Proapo-A-II which contains 2 arginine residues could be readily differentiated from mature apo-A-II which contains no arginine. The pI of proapo-A-II is 6.79, whereas the pI of the major apo-A-II isoform in plasma and lymph is 4.90. Minor apo-A-II isoforms have pI values of 5.17, 4.68, 4.42, and 4.20, respectively. Sialoisoforms of apo-A-II were identified, which had a higher apparent molecular weight on sodium dodecyl sulfate-gel electrophoresis than the major isoform and disappeared following neuraminidase treatment. The relative quantity of proapo-A-II was relatively constant in lymph very low density lipoproteins, lymph high density lipoproteins, and plasma high density lipoproteins, whereas the sialoforms and the other minor isoforms of apo-A-II were greater in lymph very low density lipoproteins and the lowest in plasma high density lipoproteins. PMID- 2981845 TI - Effects of perturbants on the pink (reduced) active form of uteroferrin. Phosphate-induced anaerobic oxidation. AB - The binuclear iron cluster of uteroferrin in its reduced and enzymatically active pink form is sensitive to a variety or perturbants. Orthophosphate, in the presence or absence of oxygen, rapidly shifts the absorption maximum of pink uteroferrin from 510 to 545 nm, concurrently abolishing the protein's g'av = 1.74 EPR signal. Apparently, therefore, dioxygen is not required for phosphate-induced oxidation of the pink protein's ferrous iron. Pyrophosphate and arsenate produce changes which differ only in degree from those induced by phosphate, suggesting that all of these structurally similar competitive inhibitors bind to a common site. Molybdate, an inhibitor even more potent than phosphate, quantitatively converts the rhombic EPR signal of pink uteroferrin into an axial signal that remains invariant to subsequent additions of phosphate. Thus, there can be inhibition without oxidation, as further evidenced by the complex EPR spectrum of undiminished intensity produced by sulfate. Fluoride, too, induces an axial component in the EPR signal of pink uteroferrin, but at high concentration abolishes the signal entirely. Vanadate also drives the protein to its oxidized, EPR-silent state, serving as an electron acceptor itself to yield the characteristic g' = 2 signal of the vanadyl (VO2+) cation. Remarkably, however, the protein remains pink, demonstrating a dissociation between color and oxidation state. Guanidinium, in contrast, causes a sizeable red shift in the pink protein's absorption maximum without loss of EPR signal intensity, showing dissociation of color and oxidation state in a complementary way. PMID- 2981846 TI - A bicarbonate-dependent process inhibitable by disulfonic stilbenes and a Na+/H+ exchange mediate 22Na+ uptake into cultured bovine corneal endothelium. AB - 22Na+ uptake into confluent monolayers of cultured bovine corneal endothelial cells was studied in the presence of ouabain (10(-4)M) to inhibit active sodium extrusion. In bicarbonate saline, uptake was reduced to a similar degree either by amiloride (10(-3)M) or by 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) (10(-3)M). A further reduction was obtained with SITS-pretreated cells in the presence of amiloride. SITS-sensitive uptake was further characterized in saline containing both ouabain (10(-4)M) and amiloride (10( 3)M). It was absolutely dependent on bicarbonate, which could not be substituted by other plasma membrane permeable buffers (50 mM acetate or 25 mM glycodiazine). It was a saturable function of both bicarbonate and sodium concentration. Half maximal fluxes occurred between 3 and 7 mM HCO3 (at 151 mM Na) and between 35 and 60 mM Na (at 28 mM HCO3). Uptake into sodium-depleted cells was reduced as opposed to sodium-rich cells, and SITS-sensitive 22Na+ efflux out of 22Na+-loaded cells into sodium-free medium was less than efflux into sodium saline, indicating trans-stimulation by sodium. The amiloride-sensitive pathway was studied in the absence of bicarbonate to inhibit uptake via the SITS-sensitive pathway. 22Na+ uptake into sodium-depleted cells increased steeply with extracellular pH in the range between pH 6 and 8 and could be largely blocked by 10(-3), but not by 10( 5) M amiloride. It is concluded that bovine corneal endothelial cells possess at least two distinct pathways for sodium uptake, amiloride sensitive 22Na+ fluxes being mediated by a Na+/H+ antiport, while the SITS-sensitive process is probably identical to a bicarbonate-sodium cotransport system postulated earlier from electrophysiological studies. PMID- 2981847 TI - Proton-activated rubidium transport catalyzed by the sodium pump. AB - Although the sodium pump normally exchanges three sodium for two potassium ions, experiments with inside-out red cell membrane vesicles show that the stoichiometry is reduced when the cytoplasmic sodium concentration is decreased to less than 1 mM. The present study was designed to gain insight into the question whether other monovalent cations, particularly protons, can act as sodium congeners in effecting pump-mediated potassium transport (ATP-dependent rubidium efflux from inside-out vesicles). The results show that at low cytoplasmic sodium concentration, an increase in proton concentration effects a further reduction in sodium:rubidium stoichiometry, to a value less than the minimal expected (1Na+:3Rb+). Furthermore, when vesicles containing 86RbCl are incubated in nominally sodium-free medium. ATP-dependent net rubidium efflux (normal influx) occurs when the pH is reduced from approximately 7.0 to 6.2 or less. This efflux is inhibited by strophanthidin and vanadate. These experiments support the notion that the sodium pump can operate as an ATP-dependent proton activated rubidium (potassium) pump without obligatory countertransport of sodium ions. PMID- 2981848 TI - Effect of P-450scc inhibitors on corticosterone production by rat adrenal cells. AB - Suspensions of rat adrenocortical cells produce corticosterone as the major glucocorticoid. Cholesterol side-chain cleavage, the initial and rate-limiting step in the glucocorticoid biosynthetic pathway, is catalyzed by P-450scc. We have examined the effect of a variety of P-450scc inhibitors on corticosterone production by isolated rat adrenocortical cells. These inhibitors include reversible, noncovalently interacting inhibitors as well as mechanism-based inhibitors which irreversibly inactivate P-450scc in vitro. (20S)-22-nor-22 thiacholesterol and (22R)-22-aminocholesterol cause 50% inhibition of corticosterone production at 4 microM and 30 nM, respectively. Inhibition by these compounds was essentially not time-dependent. (20R)-20-(1-hexynyl)-pregn-5 en-3 beta, 20-diol and (20R)-20-(1,5-hexdiynyl)-pregn-5-en-3 beta, 20-diol at 10 microM inhibited corticosterone production in a time-dependent manner, resulting in 30% inhibition of corticosterone production during a 100-min incubation. (20S) 20-(2-trimethylsilyl ethyl)-pregn-5-en-3 beta, 20-diol inhibited in a strongly time-dependent manner. At 10 microM this compound irreversibly inhibited more than 90% of the side-chain cleavage capacity of the cell during a 40-min incubation. Cells treated with this steroid did not regain their capacity for side-chain cleavage after removal of free steroid. None of the inhibitors described above inhibited production of corticosterone by cells supplied with pregnenolone, the product of the P-450scc reaction. We suggest that the only significant effect of these compounds under these conditions is inhibition of the side-chain cleavage enzyme. PMID- 2981849 TI - Characterization of the ribosomal RNA gene clusters in Halobacterium cutirubrum. AB - We present a comprehensive and detailed analysis of the structure and organization of a cloned ribosomal RNA gene cluster from the archaebacterial species Halobacterium cutirubrum. With the exception of a region in the middle of the 23 S rRNA gene, the DNA sequence of the entire gene cluster has been determined. The gene organization is similar to that found in typical eubacteria with the 16, 23, and 5 S genes occupying the proximal, middle, and distal positions, respectively. There appears to be no equivalent to the eucaryotic 5.8 S gene in H. cutirubrum. The cluster also contains two putative tRNA genes, an alanine tRNA gene in the 16-23 S intergenic space, and a cysteine tRNA gene distal to the 5 S rRNA gene. The 16 and 23 S rRNA genes are surrounded by long nearly perfect inverted repeat sequences which are presumably utilized along with other structural features of the RNA for the processing of 16 and 23 S rRNA from a large precursor transcript. The 5' sequence flanking the 16 S rRNA gene contains two imperfect copies, followed by three perfect copies of a bipartite direct-repeat unit. The sequence AAGTAA, believed to be an important component of the Halobacterium promotor, is present in the highly conserved portion of the direct repeat unit. In the 3' region flanking the 5 S rRNA gene there are sequences, a short inverted repeat followed by T5, and a G/C-rich region followed by an A/T-rich region, which may function in transcription termination. Genomic southern hybridization experiments clearly indicate that the ribosomal RNA genes are unique single-copy DNA in H. cutirubrum. PMID- 2981850 TI - The structural organization of the chicken calmodulin gene. AB - The structural organization of the entire chicken calmodulin (CaM) gene was determined by analysis of overlapping genomic clones obtained from Charon 4A and cosmid DNA libraries. These clones together span 39 kilobases of chicken genomic DNA. The CaM gene is 12 kilobases long and contains 8 exons interrupted by introns of highly variable size. The first intron (A) is only 19 base pairs (bp) long and divides the 5' untranslated region. Intron B separates the ATG from the first nucleotide of the triplet which encodes the NH2-terminal amino acid of CaM (Ala) whereas intron C separates the triplets encoding amino acids 10-11. Introns D, F, and G interrupt the Ca2+ binding subdomains II, III, and IV of CaM whereas intron E is localized in the linker region between the highly homologous NH2- and COOH-terminal halves of the protein. Primer extension studies using chicken brain poly(A+) mRNA and a fragment from the 5' untranslated region of a CaM cDNA identified the presumptive transcription initiation site (cap site) of the mRNA to be 103 bp 5' from the initiation condon ATG. A consensus sequence TATTTAA was localized 29 bp 5' from this cap site while a CCAAT sequence was located further 5' at position -58 bp. The structure of the CaM gene is strikingly similar to genes that encode other Ca2+ binding proteins from sea urchin, chicken, rat, and quail. These data suggest a conservation of genome organization related to the calcium binding and regulatory domains of Ca2+ binding proteins. PMID- 2981851 TI - Increased superoxide radical production evokes inducible DNA repair in Escherichia coli. AB - Paraquat induced the SOS response in Escherichia coli. This was measured in terms of acquired resistance towards UV lethality in a wild-type strain and in terms of appearance of beta-galactosidase activity in a din::Mu d(Ap lac) fusion strain. However measured, the induction of the SOS response by paraquat was entirely dioxygen-dependent; whereas induction of the SOS response by mitomycin C was independent of the presence of dioxygen. As expected, recA(Def) and lexA(Ind-) isogenic strains did not show the SOS response. It appears likely that O-2, whose intracellular production is increased by paraquat, leads to DNA damage which in turn induces the SOS response. PMID- 2981852 TI - Characterization of a transferrin-diphtheria toxin conjugate. AB - We report here the synthesis and properties of a hybrid toxin prepared by covalently coupling diphtheria toxin to transferrin. The purified material contained two major hybrid protein species and was highly cytotoxic to mouse LMTK cells in culture, reducing protein synthesis by 50% in 24 h at a concentration of 1 ng/ml. Cytotoxic activity was completely abolished in the presence of exogenous transferrin or anti-transferrin or anti-diphtheria toxin, thus demonstrating that the hybrid toxin was intoxicating cells via their transferrin receptors and that both the diphtheria toxin and transferrin components of the conjugate were necessary for activity. NH4Cl, a drug that elevates the pH within acidic intracellular vesicles, also blocked cytotoxic activity, suggesting that a low intravesicular pH was required for activity. The inhibitory effect of NH4Cl could be abolished by exposing toxin-treated cells to acidic culture medium, further implicating an acid-dependent step in the mechanism of the hybrid toxin action. Studies on the kinetics of intoxication also implied that endocytosis and exposure to a low pH within vesicles were necessary for cytotoxicity. Altogether, the results suggest that the transferrin-diphtheria toxin conjugate binds to transferrin receptors and is internalized into acidic endocytic vesicles. The enzymatic moiety of diphtheria toxin then apparently enters the cytosol in response to the low pH and subsequently arrests protein synthesis. PMID- 2981853 TI - Inositol 1,4,5-trisphosphate releases Ca2+ from a Ca2+-transporting membrane vesicle fraction derived from human platelets. AB - Human platelet membrane vesicles that accumulated Ca2+ in the presence of ATP were isolated on an isoosmotic KCl-Percoll gradient. ATP-dependent Ca2+ uptake was stimulated by oxalate and phosphate to steady-state levels of greater than 100 nmol/mg protein, and the accumulated Ca2+ could be largely released by ionophore A23187. Inositol 1,4,5-trisphosphate, in a dose-dependent manner (0.5 5.0 microM), caused the rapid release (less than 5 s) of 40-70% of the total A23187-releasable store of accumulated Ca2+. The membrane vesicles that release accumulated Ca2+ in response to inositol 1,4,5-trisphosphate were enriched in enzymes characteristically found in smooth endoplasmic reticulum. These results support the hypothesis that inositol 1,4,5-trisphosphate, produced by the hydrolysis of phosphatidylinositol 1,4-bisphosphate in response to stimulation of cell surface receptors, is a second messenger mediating the release of Ca2+ from intracellular storage sites. PMID- 2981854 TI - Interaction and identification of ubiquinone-binding proteins in ubiquinol cytochrome c reductase by azido-ubiquinone derivatives. AB - Various azido-ubiquinone derivatives were synthesized and characterized. 3-Azido 2-methyl-5-methoxy-6-(3,7-dimethyloctyl)-1,4-benzoquinone was found to be suitable for the study of specific interaction between ubiquinone (Q) and protein. It was synthesized with high specific radioactivity and used to identify the Q-binding proteins in purified ubiquinol-cytochrome c reductase. This azido-Q derivative showed partial efficiency in restoring activity to the Q- and phospholipids-depleted ubiquinol-cytochrome c reductase in the absence of light. Azido-Q derivative treated samples, however, became completely inactivated upon photolysis, and the inactivation was not reversed by addition of Q derivatives. The redox state of the azido-Q derivative has little effect on the Q-binding affinity. Two protein subunits with Mr = 37,000 and 17,000 were found to be heavily labeled when depleted ubiquinol-cytochrome c reductase was treated with [3H] azido-Q derivative followed by photolysis and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The amount of radioactive labeling of the Mr = 17,000 protein was proportional to the degree of inactivation and affected by the presence of phospholipids. The radioactive labeling of the Mr = 37,000 protein subunit, however, showed no correlation with degree of inactivation and was not affected by phospholipids. Since the radiolabeling at the Mr = 17,000 protein subunit was affected by phospholipids and correlated with the enzymatic activity, this subunit is probably the Q-binding protein in this enzyme complex (QPc). The inhibition of enzymatic activity by n-heptyl-4-hydroxyquinoline-N oxide was easily reversed by addition of the azido-Q derivative. The distribution of radioactivity among the subunits of ubiquinol-cytochrome c reductase was not affected by the presence of antimycin A, 5-n-undecyl-6-hydroxy-4,7 dioxobenzothiazole or n-heptyl-4-hydroxyquinoline-N-oxide, suggesting that the binding site(s) of these inhibitors are not the Q-binding site. PMID- 2981855 TI - Respective role of each of the purine N-6 amino groups of 5'-O triphosphoryladenylyl(2'----5')adenylyl(2----5')adenosine in binding to and activation of RNase L. AB - We have synthesized a series of 2-5A (ppp5'-A2'p5'A2'p5'A) analogs in which each adenosine residue has been sequentially replaced by inosine: viz., ppp5'I2'p5'A2'p5'A, ppp5'A2'p5'I2'p5'A, and ppp5'A2'p5'A2'p5'I. These transformations enabled us to delineate the role of each of the three purine N-6 amino groups of 2-5A in determining oligonucleotide binding to and activation of the 2-5A-dependent endoribonuclease, RNase L. With the RNase L activity of both mouse L cells and human Daudi lymphoblastoid cells, we found that the N-6 amino group of the first adenosine nucleotide residue (from the 5'-terminus) is of crucial importance in determining binding to the endonuclease; however, removal of the N-6 amino moieties of the second or third adenosine nucleotide residues resulted in only a minimal decrease in binding to the endonuclease. On the other hand, conversion of the third adenosine residue to inosine effected a dramatic (10,000-fold compared to 2-5A) loss in ability to activate the nuclease; however, execution of the same N-6 amino group conversion at either the first or second adenosine residue did not cause a major change in nuclease activation ability when the accompanying decreased endonuclease binding was considered. These results clearly demonstrate that the N-6 amino group of the first adenosine residue of 2-5A is critical in RNase L binding whereas the N-6 amino function of the third adenosine residue of 2-5A is crucial for the activation of RNase L. PMID- 2981856 TI - Reversible acidic-alkaline transition of the carbon monoxide complex of cytochrome c peroxidase. AB - The Soret absorption band of the ferrous carbon monoxide (CO) complex of cytochrome c peroxidase exhibited a blue shift from 423.7 to 420 nm upon an increase in pH from 6.5 to 8.5. The spectral change was reversible with an isosbestic point at 422 nm. The pH dependence of this spectral change gave a sigmoidal curve fitted well to a theoretical curve of a cooperative release of two protons with a pK value of 7.5, indicating the existence of the acidic and alkaline forms of the ferrous CO enzyme. Upon irradiation of light flash (100 J of power and 30-microseconds), the heme-bound CO was readily dissociated in both acidic and alkaline forms with a quantum yield of approximately unity. On the other hand, the rate of recombination of the dissociated CO with the heme iron was significantly different between these two forms; the recombination rate constants were 1.1 X 10(3) and 3.0 X 10(4) M-1 S-1 at 25 degrees C for the acidic and alkaline forms, respectively. At intermediate pH values, kinetics of recombination were biphasic, consisting of the slow and fast processes with the appropriate rate constants mentioned above. When the fraction of the fast process was plotted against pH, the pH profile coincided with the spectrophotometric pH titration curve described above. Thus, it was concluded that the acidic and alkaline forms of the enzyme were responsible for the slow and fast processes, respectively. In infrared spectroscopy, the acidic form showed a narrow CO stretching band at 1922 cm-1 with a half-band width of 12.5 cm-1, while the alkaline form exhibited a broad CO-stretching band at 1948 cm-1 with a half-band width of 33 cm-1. Significance of these results are discussed in relation to the structure of the heme vicinity on the CO complex of cytochrome c peroxidase. PMID- 2981857 TI - Na/K/Cl cotransport in cultured human fibroblasts. AB - The transport characteristics and regulation of the Na/K/Cl cotransport system were investigated in cultured human fibroblasts (HSWP cells). The existence of the system was documented by the finding that digitoxin-insensitive K+ influx was dependent upon the presence of both Na+ and Cl- in the extracellular milieu. It was found that only Br- could partially substitute for Cl-, with SCN-, I-, acetate, and gluconate being ineffective. Li+ could partially substitute for Na+; however, choline was without effect. The shape of the titration curves for K+ influx versus extracellular Cl- concentration was dependent upon the substituted anion. Furthermore, the apparent Km for Cl- at saturating [K+]o and [Na+]o, was also dependent upon the substituted anion and ranged from 30 mM (gluconate substitution) to 100 mM (acetate substitution). The titration curves for K+ influx versus extracellular Na+ concentration displayed hyperbolic kinetics and the apparent Km = 15 mM at saturating [K+]o. The curve for K+ influx versus extracellular K+ concentration was a hyperbola and the apparent Km for K+ = 3 mM at saturating [Na+]o. The digitoxin-insensitive K+ flux was found to be sensitive to related 5-sulfamoylbenzoic acid derivatives, commonly known as "loop" diuretics and to be insensitive to both: amiloride (3,5-diamino-N (aminoiminomethyl)-6-chloropyrazinecarboxamide++ +) and 4-acetamido-4' isothiocyanostilbene-2,2'-disulfonic acid. The Na/K/Cl cotransport system was not stimulated by serum, but was slightly stimulated by two peptide mitogens. Furthermore, agents which cause an elevation in cellular cyclic AMP levels were found to be potent inhibitors of cotransport. PMID- 2981858 TI - Specificity of the functional interactions of the beta-adrenergic receptor and rhodopsin with guanine nucleotide regulatory proteins reconstituted in phospholipid vesicles. AB - We have assessed the functional interactions of two pure receptor proteins with three different pure guanine nucleotide regulatory proteins in phosphatidylcholine vesicles. The receptor proteins are the guinea pig lung beta adrenergic receptor (beta AR) and the retinal photon receptor rhodopsin. The guanine nucleotide regulatory proteins were the stimulatory (Ns) and inhibitory (Ni) proteins of the adenylate cyclase system and transducin (T), the regulatory protein from the light-activated cyclic GMP phosphodiesterase system in retinal rod outer segments. The insertion of Ns with beta AR in lipid vesicles increases the extent of binding of [35S] GTP gamma S to Ns and in parallel, the total GTPase activity. However, there is little change in the actual rate of catalytic turnover of GTPase activity (defined as mol of Pi released/min/mol of Ns-guanine nucleotide complexes). Enhancement of this turnover rate requires the beta agonist isoproterenol and is accounted for by an isoproterenol-promoted increase in the rate and extent of [35S]GTP gamma S binding to Ns. The co-insertion of the beta AR with Ni or transducin results in markedly lower stimulation by isoproterenol of both the GTPase activity and [35S]GTP gamma S binding to these nucleotide regulatory proteins indicating that their preferred order of interaction with beta AR is Ns much greater than Ni greater than T. This contrasts with the preferred order of interaction of these different nucleotide regulatory proteins with light-activated rhodopsin which we find to be T approximately equal to Ni much greater than Ns. Nonetheless the fold stimulation of GTPase activity and [35S]GTP gamma S binding in T, induced by light-activated rhodopsin, is significantly greater than the "fold" stimulation of these activities in Ni. This reflects the greater intrinsic ability of Ni to hydrolyze GTP and bind guanine nucleotides (at 10 mM MgCl2, 100-200 nM GTP or [35S] GTP gamma S) compared to T. The maximum turnover numbers for the rhodopsin-stimulated GTPase in both Ni and T are similar to those obtained for isoproterenol stimulated activity in Ns. This suggests that the different nucleotide regulatory proteins are capable of a common upper limit of catalytic efficiency which can best be attained when coupled to the appropriate receptor. PMID- 2981859 TI - Assembly of the mitochondrial membrane system. CBP6, a yeast nuclear gene necessary for synthesis of cytochrome b. AB - A new gene function involved in the expression of mitochondrial cytochrome b is described. E158 is a respiratory deficient strain of Saccharomyces cerevisiae with a recessive mutation in nuclear DNA. The block in mitochondrial respiration is a consequence of the mutant's inability to synthesize cytochrome b. This has been confirmed by analysis of the mitochondrial translation products in E158. Since the mutant has wild-type concentrations of mature cytochrome b mRNA, the absence of the cytochrome b protein cannot be due to a transcriptional or RNA processing defect. The wild-type gene (CBP6) responsible for the observed phenotype has been cloned by transformation of E158 with a genomic library of yeast nuclear DNA. The cloned gene has been sequenced and shown to code for a basic protein with a molecular weight of 18,657. Both deletion and disruption of the CBP6 coding sequence in chromosomal DNA of wild-type yeast lead to respiratory deficiency with a concomitant loss of cytochrome b. Part of the CBP6 gene has been fused to the trpE gene on a high expression Escherichia coli vector. The hybrid protein encoded by the trpE/CBP6 fusion has been purified from E. coli and used as an antigen for antibody production. Antibodies to the hybrid protein cross-react with an 18-kDa protein present in yeast mitochondria. PMID- 2981860 TI - APS kinase from Penicillium chrysogenum. Dissociation and reassociation of subunits as the basis of the reversible heat inactivation. AB - Adenosine-5'-phosphosulfate (APS) kinase from Penicillium chrysogenum, loses catalytic activity at temperatures greater than approximately 40 degrees C. When the heat-inactivated enzyme is cooled to 30 degrees C or lower, activity is regained in a time-dependent process. At an intermediary temperature (e.g. 36 degrees C) an equilibrium between active and inactive forms can be demonstrated. APS kinase from P. chrysogenum is a dimer (Mr = 57,000-60,000) composed of two apparently identical subunits. Three lines of evidence suggest that the reversible inactivation is a result of subunit dissociation and reassociation. (a) Inactivation is a first-order process. The half-time for inactivation at a given temperature is independent of the original enzyme concentration. Reactivation follows second-order kinetics. The half-time for reactivation is inversely proportional to the original enzyme concentration. (b) The equilibrium active/inactive ratio at 36 degrees C increases as the total initial enzyme concentration is increased. However, Keq,app at 5 mM MgATP and 36 degrees C calculated as [inactive sites]2/0.5 [active sites] is near-constant at about 1.7 X 10(-8) M over a 10-fold concentration range of enzyme. (c) At 46 degrees C, the inactive P. chrysogenum enzyme (assayed after reactivation) elutes from a calibrated gel filtration column at a position corresponding to Mr = 33,000. Substrates and products of the APS kinase reaction had no detectable effect on the rate of inactivation. However, MgATP and MgADP markedly stimulated the reactivation process (kapp = 3 X 10(5) M-1 X s-1 at 30 degrees C and 10 mM MgATP). The kapp for reactivation was a nearly linear function of MgATP up to about 20 mM suggesting that the monomer has a very low affinity for the nucleotide compared to that of the native dimer. Keq,app at 36 degrees C increases as the MgATP concentration is increased. The inactivation rate constant increased as the pH was decreased but no pK alpha could be determined. The reactivation rate constant increased as the pH was increased. An apparent pK alpha of 6.4 was estimated. PMID- 2981861 TI - A variant form of the 1,25-dihydroxyvitamin D3 receptor with low apparent hormone affinity in cultured monkey kidney cells (LLC-MK2). A model for tissue resistance to vitamin D. AB - Chandler et al. (Chandler, J.S., Chandler, S.K., Pike, J.W., and Haussler, M.R. (1984) J. Biol. Chem. 259, 2214-2222) previously demonstrated that 1,25 dihydroxyvitamin D3 (1,25-(OH)2D3) caused the induction of 25-hydroxyvitamin D3 24-hydroxylase (24-hydroxylase) in a rhesus monkey kidney cell line (LLC-MK2) apparently deficient in the high affinity 1,25-(OH)2D3 receptor. We have re examined this phenomenon and report here that 24-hydroxylase induction is mediated by a receptor variant in LLC-MK2 cells with low hormone affinity. Dose response analysis showed that in contrast to LLC-PK1 (a typical receptor-positive cell line), the LLC-MK2 line was less sensitive to 1,25-(OH)2D3 by 2 orders of magnitude. Employing optimal concentrations of 1,25-(OH)2D3 for 24-hydroxylase induction in each cell type, the early time courses of this bioresponse were identical in LLC-MK2 and LLC-PK1 and were consistent with a nuclear action of hormone-receptor complexes. Moreover, the rank order of potency of vitamin D3 congeners as inducers of 24-hydroxylase activity in LLC-MK2 cells agreed well with their relative affinity for the 1,25-(OH)2D3 receptor. An examination of 1,25-(OH)2D3 receptor content via DNA-cellulose chromatography in LLC-MK2 cells incubated at ligand concentrations 10-25-fold higher than the normal 2 nM revealed a minimum of 1600 receptor-like molecules/LLC-MK2 cell. These results show that LLC-MK2 cells possess a variant receptor form with apparent low affinity for 1,25-(OH)2D3. This system should serve as a model for clinical syndromes characterized by the requirement for massive doses of vitamin D to prevent rickets. PMID- 2981862 TI - sn-1,2-Dioctanoylglycerol. A cell-permeable diacylglycerol that mimics phorbol diester action on the epidermal growth factor receptor and mitogenesis. AB - The cell-permeable diacylglycerol, sn-1,2-dioctanoylglycerol (DiC8), is shown to mimic the effect of tumor promoting phorbol diesters on epidermal growth factor (EGF) binding and action in intact cells. DiC8 inhibited the binding of [3H]phorbol dibutyrate to A431 cell monolayers indicating that the diacylglycerol interacts with the phorbol diester receptor. At 0.3 microM, DiC8 half-maximally inhibited the high affinity binding of 125I-EGF to A431 human epidermoid carcinoma cells. Scatchard analysis indicated that the inhibition of 125I-EGF binding was very similar to that observed in the presence of 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA). DiC8 also mimicked the action of PMA to increase the phosphorylation state of the EGF receptor in 32P-labeled cells. Phosphoamino acid analysis demonstrated that DiC8 and PMA caused an increase in the level of EGF-receptor phosphoserine and phosphothreonine, whereas EGF caused an increase in the level of phosphoserine, phosphothreonine, and phosphotyrosine. Phosphopeptide mapping of the EGF receptor showed that DiC8 and PMA enhanced the phosphorylation of the same tryptic peptides. DiC8 inhibited the EGF-dependent tyrosine phosphorylation of the EGF receptor in A431 cells in a similar manner to that observed with PMA. In further experiments with quiescent Swiss 3T3 fibroblasts, DiC8 mimicked the ability of PMA to stimulate the incorporation of [methyl-3H]thymidine synergistically with low concentrations of EGF. This result indicates that DiC8 will mimic the long-term effects of PMA to regulate mitogenesis and raises the possibility that it may be active in two stage carcinogenesis. As both DiC8 and PMA stimulate the Ca2+- and phospholipid dependent protein kinase (C-kinase) in vitro, the results support the hypothesis that the activation of C-kinase is a critical component of phorbol diester action on EGF receptor modulation and cell proliferation. PMID- 2981863 TI - Phosphorylation of sites 3 and 4 in rabbit skeletal muscle glycogen synthase by cAMP-dependent protein kinase. AB - Rabbit skeletal muscle glycogen synthase (synthase a) can be phosphorylated by 0.2 to 2.5 microM catalytic subunit of cAMP-dependent protein kinase to a stoichiometry of 1.5 to 3 mol of 32PO4/subunit (90,000 X g). When a complete tryptic digest of this 32P-synthase was chromatographed on reverse phase high performance liquid chromatography, it was observed that, in addition to sites 1a, 1b, and 2, site 3 was phosphorylated. The peptide containing site 5 also contained 32PO4, but sequence analysis identified a new phosphorylation site, site 4 (Arg-His-Ser-Ser(PO4)-) which precedes site 5. Phosphorylation of sites 3 and 4 became significant when the total phosphorylation stoichiometry exceeded 1.5 mol/subunit. The heat-stable protein kinase inhibitor protein or the regulatory subunit of cAMP-dependent protein kinase decreased the phosphorylation of all sites, including sites 3 and 4. Phosphorylation of all sites by the holoenzyme form of cAMP-dependent kinase was highly dependent on the presence of cAMP. These results establish that phosphorylation of these sites is due to the cAMP-dependent protein kinase itself and not to a contaminating kinase(s). Synthase b from control rabbit muscle, containing 2 to 2.5 mol of phosphate/subunit, incorporated up to 2.0 mol of 32PO4 catalyzed in vitro by the cAMP-dependent protein kinase. Again, significant 32PO4 was detected in sites 3 and 4 as well as in 1a, 1b, and 2. These results suggest that the in vivo phosphorylation of sites 1a, 1b, 2, and 3 observed after injection of epinephrine in rabbits could be due solely to the known activation of the cAMP-dependent protein kinase. PMID- 2981864 TI - Functional heterogeneity of ubiquitin carrier proteins. AB - In the formation of covalent ubiquitin-protein conjugates that occurs during ATP- and ubiquitin-dependent proteolysis in reticulocyte extracts, ubiquitin (Ub) is activated to a thiol ester of the activating enzyme E1 (via the Ub carboxyl terminus), transferred to low-molecular weight "carrier proteins" (E2s) to form E2-Ub thiol esters, and then transferred by a third enzyme (E3) to amino groups on target proteins (Hershko, A., Heller, H., Elias, S., and Ciechanover, A. (1983) J. Biol. Chem. 258, 8206-8214). We report here the fractionation of Ub carrier proteins by molecular weight, and their characterization with respect to several activities. The Ub thiol ester forms of at least four of the five E2s catalyze Ub transfer to a number of small amines, in a reaction that does not require E3; only primary amines on primary carbons can serve as Ub acceptors. E3 independent Ub transfer to the small, basic proteins histones H2A and H2B, and cytochrome c, is also observed. The Ub thiol ester forms of two of the E2s were found to catalyze Ub transfer to cytochrome c. Only a single E2 functions in E3 dependent conjugate formation (with the substrates creatine phosphokinase, reduced/carboxymethylated serum albumin, and oxidized RNase) and in E3-dependent protein breakdown (with the substrate serum albumin). This E2 has a subunit molecular weight of 14,000 and migrates as a dimer on Sephacryl 200. PMID- 2981865 TI - Characterization of tyrosyl kinase activity in human serum. AB - Tyrosyl kinase activity was detected in 1.0 microliter or less of human serum with the substrates angiotensin II, polyamino acid polymer Glu-Tyr (4:1), anti pp60src IgG, and endogenous serum proteins. Most (about 84%) of the tyrosyl kinase activity was in the 100,000 X g soluble fraction from serum and a similar level of activity was found in the soluble fraction from plasma. Expression of tyrosyl kinase activity in individual serum samples differed more than 15-fold. The different levels of tyrosyl kinase activity were not likely due to phosphatases, proteases, ATPases, or kinase inhibitors and activators in serum. The normal serum and plasma tyrosyl kinase activities were not stimulated by epidermal growth factor, platelet-derived growth factor, insulin, or growth factors from fetal calf serum. Investigations of samples from patients with malignant disorders indicated that those with malignant melanoma contained the highest levels of serum tyrosyl kinase activity. PMID- 2981866 TI - Studies on the hepatic alpha 1-adrenergic receptor. Modulation of guanine nucleotide effects by calcium, temperature, and age. AB - The effects of guanine nucleotides on the hepatic alpha 1-adrenergic receptor were studied using norepinephrine (NE) displacement of [3H]prazosin binding to rat liver plasma membranes. Nonhydrolyzable GTP analogues caused large rightward shifts of norepinephrine displacement curves of [3H]prazosin binding in EGTA treated membranes, but only small shifts in membranes prepared with Ca2+. The effect of a brief Ca2+ exposure on NE displacement curves was not reversed by adding excess EGTA prior to binding experiments. Analysis of the curves showed that the EGTA membranes had an increased number of high affinity agonist sites (Kd, 42 nM) and that guanyl-5'-yl imidodiphosphate (GppNHp) converted these to low affinity sites (Kd, 1039 nM). When binding was carried out at 2 degrees C, the norepinephrine displacement curves were shifted to the left, and GppNHp was without effect. Neither EGTA, Ca2+, nor 2 degrees C treatment altered [3H]prazosin binding per se. Attempts were made to differentiate the potency order of GTP analogues which alter glucagon receptor binding (presumably mediated by the stimulatory GTP-binding protein, Na, of the adenylate cyclase system) from the potency order of GTP analogues which alter alpha 1-receptor agonist binding (presumably mediated by a yet uncharacterized GTP-binding protein which some have speculated may be distinct from Ns). However, the potency series of GTP analogues to alter norepinephrine binding was GTP gamma S greater than GppNHp greater than or equal to GTP greater than or equal to GDP greater than or equal to GppCHp greater than GMP (where GTP gamma S represents guanosine 5'-O-(thiotriphosphate) and GppCHp represents guanyl-5'-yl (beta, gamma-methylene)diphosphonate) and was identical to that for inhibition of [125I]iodoglucagon binding. The ability of GppNHp to alter norepinephrine displacement of [3H]prazosin binding increased with the age of the rat from which membranes were prepared. This was due to the fact that juvenile rats (50-75 g) had few alpha 1-receptors in the high affinity state, whereas in old rats (430-490 g) more of the receptors were in this form. Age has previously been shown to increase alpha 1-adrenergic stimulation of cAMP in isolated hepatocytes (Morgan, N.G., Blackmore, P. F., and Exton, J. H. (1983) J. Biol. Chem. 258, 5103-5109) but did not affect the dose-response curves for norepinephrine-induced Ca2+ mobilization and phosphorylase activation in these cells. These data suggest that alpha 1-adrenergic receptors can become coupled to a guanine nucleotide-responsive moiety in hepatic plasma membranes and that this may be similar to Ns.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981867 TI - Effects of estrogen on apolipoprotein secretion by the human hepatocarcinoma cell line, HepG2. AB - We have examined the effect of estrogen on the rate of accumulation of apolipoproteins secreted by the human hepatocarcinoma cell line, HepG2. Prior to exposure to hormone, we detected less than 300 high-affinity, nuclear, estrogen binding sites/cell. Within 48 h of growth in the presence of 20 nM 17 beta estradiol this number rose to 3000-3500 sites/cell. Rates of accumulation of two of the major apolipoproteins, apo-C-II and apo-A-I increased 2.5- and 2.0-fold, respectively, in response to estrogen treatment. Other major apolipoproteins were not affected at this concentration of hormone. Induction of both proteins was completely antagonized by 20 nM testosterone. The density distribution of apolipoproteins secreted by the hepatocytes was similar to that reported using perfused liver systems. The consequences of estrogen treatment were to increase the apo-C-II/apo-C-III ratio in very low density lipoproteins as well as to decrease the overall very low density lipoprotein:high density lipoprotein ratio. PMID- 2981868 TI - Apolipoprotein C-II mRNA levels in primate liver. Induction by estrogen in the human hepatocarcinoma cell line, HepG2. AB - We have demonstrated that physiological concentrations of 17 beta-estradiol increase nuclear estrogen-specific binding sites in the human hepatoma cell line HepG2 7- to 10-fold and the rate of accumulation of secreted apolipoprotein C-II (apo-C-II), 2.5-fold (Tam, S-P., Archer, T. K., and Deeley, R. G. (1985) J. Biol. Chem. 260, 1670-1675). Apo-C-II is the major activator of lipoprotein lipase, an enzyme which plays a key role in lipoprotein catabolism. In order to define more precisely the mechanism by which estrogen influences apo-C-II production, we have synthesized a triacontanucleotide DNA probe that is complementary to apo-C-II mRNA. We have used the probe both in Northern hybridization experiments and in DNA excess titrations to quantify apo-C-II mRNA in hormonally treated HepG2 cells and various primate tissues. These studies revealed that: 1) the concentration of apo-C-II mRNA in HepG2 cells is comparable with that present in human liver; 2) treatment of the cells with low levels of estrogen results in a doubling of the apo-C-II mRNA concentration; 3) the apo-C-II mRNA concentration in monkey liver is 60- to 70-fold greater than in the intestine and 2.5-fold higher than in human liver. PMID- 2981869 TI - Calcium-dependent conformational changes in the 36-kDa subunit of intestinal protein I related to the cellular 36-kDa target of Rous sarcoma virus tyrosine kinase. AB - Protein I from intestinal epithelium is biochemically and immunologically related to the fibroblast 36-kDa substrate of the Rous sarcoma virus-encoded tyrosine protein kinase (Gerke and Weber (1984) EMBO J. 3, 227-233). Protein I is a Ca2+ binding protein containing two copies each of a 36- and 10-kDa subunit. Denaturation/renaturation experiments show that the 36-kDa subunit is a monomer, whereas the 10-kDa subunit forms a dimer. Mixing of the subunits leads to reconstituted protein I. Physicochemical properties of protein I and its isolated subunits reveal a Ca2+-dependent conformational change in the 36-kDa subunit which involves the exposure of 1 or more tyrosine residues to a more aqueous environment. This change points to a Ca2+ binding constant of about 10(4) M-1 in the presence of 2 mM Mg2+ and induces the ability of protein I and the 36-kDa subunit to bind in vitro to F-actin and nonerythroid spectrin. The same high Ca2+ requirement has been reported for the in vitro tyrosine phosphorylation of a 35 kDa protein from A-431 carcinoma cells by the epidermal growth factor receptor kinase (Fava and Cohen (1984) J. Biol. Chem. 259, 2636-2645). Here we show that this 35-kDa substrate is biochemically and immunologically related to the 36-kDa subunit of protein I, which in turn corresponds to the substrate of the Rous sarcoma virus kinase. The protein of A-431 cells exists not only as a monomer but also as a dimer. The latter fraction contains a 10-kDa polypeptide immunologically related to the corresponding subunit of protein I. Given past results on the A-431 system, we speculate that the monomer rather than the dimer is the preferred in vitro substrate for the epidermal growth factor receptor kinase. Thus, the 10-kDa subunit, which induces dimerization of the phosphorylatable large subunit, may act as an inhibitor. PMID- 2981870 TI - Topography of human placental receptors for epidermal growth factor. AB - These studies were undertaken to determine whether term human placental microvillus plasma membranes, which are exposed to maternal blood, and basolateral plasma membranes, which are in close proximity to fetal blood capillaries, contain receptors for epidermal growth factor (EGF). These two highly purified membranes bound 125I-EGF with similar affinity (apparent dissociation constants, 0.07-0.12 nM, but the total number of available receptors was greater in microvillus (8.2 pmol/mg protein) compared to basolateral (4.9 pmol/mg protein) plasma membranes. Detailed characterization of 125I-EGF binding to these membranes revealed numerous similarities as well as differences. The two membranes contained two major (155 and 140 kDa) and at least three minor (115, 175, and 210 kDa) specific 125I-EGF binding proteins. The 115-kDa protein was only found in basolateral plasma membranes. The 155-kDa protein was predominantly labeled in microvillus, whereas the 140-kDa protein was labeled predominantly in basolateral plasma membranes. The addition of protease inhibitors did not alter the multiple 125I-EGF binding proteins pattern found in these membranes. EGF stimulated phosphorylation of 140- and 155-kDa proteins in both microvillus and basolateral plasma membranes. However, the 155-kDa protein was phosphorylated to a greater extent in microvillus, whereas both 140- and 155-kDa proteins were phosphorylated equally in basolateral plasma membranes. Light and electron microscope autoradiographic studies revealed that 125I-EGF preferentially associated with microvillus plasma membranes. The data demonstrates the presence of EGF receptors in outer cell membranes of syncytiotrophoblasts and suggests that maternal EGF may influence syncytiotrophoblast function by binding to receptors in microvillus plasma membranes, while fetal EGF may also influence syncytiotrophoblast function but via receptors in basolateral plasma membranes. PMID- 2981871 TI - Altered triple helical structure of type I procollagen in lethal perinatal osteogenesis imperfecta. AB - Cultured dermal fibroblasts from an infant with the lethal perinatal form of osteogenesis imperfecta (type II) synthesize normal and abnormal forms of type I procollagen. The abnormal type I procollagen molecules are excessively modified during their intracellular stay, have a lower than normal melting transition temperature, are secreted at a reduced rate, and form abnormally thin collagen fibrils in the extracellular matrix in vitro. Overmodification of the abnormal type I procollagen molecules was limited to the NH2-terminal three-fourths of the triple helical domain. Two-dimensional mapping of modified and unmodified alpha chains of type I collagen demonstrated neither charge alterations nor large insertions or deletions in the region of alpha 1(I) and alpha 2(I) in which overmodification begins. Both the structure and function of type I procollagen synthesized by cells from the parents of this infant were normal. The simplest interpretation of the results of this study is that the osteogenesis imperfecta phenotype arose from a new dominant mutation in one of the genes encoding the chains of type I procollagen. Given the requirement for glycine in every third position of the triple helical domain, the mutation may represent a single amino acid substitution for a glycine residue. These findings demonstrate further heterogeneity in the biochemical basis of osteogenesis imperfecta type II and suggest that the nature and location of mutations in type I procollagen may determine phenotypic variation. PMID- 2981872 TI - Identification and cyclic AMP-induced modification of the cyclic AMP receptor in Dictyostelium discoideum. AB - We have recently identified a cell surface cAMP-binding protein by specific photoaffinity labeling of intact Dictyostelium discoideum cells with 8-N3-[32P] cAMP. The major photolabeled protein appears as a doublet (Mr = 40,000-43,000) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiography. In this study, the doublet is shown to have the characteristics of the cAMP receptor responsible for chemotaxis and cAMP signaling. Both specific photoaffinity labeling of the doublet and binding of 8-N3-[32P]cAMP are saturable (KD = 0.3 microM), the levels of both peak at 5 h, and both are inhibited by cAMP and several cAMP analogs in the same order of potency and with K1 values similar to those measured for inhibition of [3H]cAMP binding. When cAMP-binding activity was partially purified (40-fold) and then photoaffinity labeled, the same bands (Mr = 40,000-43,000) were observed. The relative intensities of the upper and lower bands of the doublet alternated at the same frequency as the spontaneous oscillations in cAMP synthesis. When oscillations were suppressed, the lower band of the doublet predominated. Following addition of cAMP, the relative intensity gradually shifted to the upper band. When cAMP was removed, there was a gradual restoration of the lower band form. We propose that the lower band form of the receptor activates chemotaxis and cAMP signaling and that the upper band form does not. This reversible receptor modification may then be the mechanism of adaptation, the process by which the physiological responses cease to be stimulated by persistent cAMP. Several developmentally regulated genes in D. discoideum have been reported to be induced or suppressed by pulses of cAMP (adaptive regulation) and others by continuous cAMP (nonadaptive regulation). These observations may be explained by the receptor modification reported here if the two forms of the receptor, which bind cAMP with the same affinity, independently influence gene expression. PMID- 2981873 TI - Cloning of the A gene of bacteriophage Mu and purification of its product, the Mu transposase. AB - The bacteriophage Mu transposase (the Mu A gene product), which is absolutely required for both integration of Mu and replicative transposition during the lytic cycle, has been overproduced by cloning the gene on a plasmid under the control of the phage lambda PL promoter. The protein has been purified to near homogeneity from the lysate of heat-induced cells of a strain carrying the plasmid. The purified protein is active as judged by its ability to complement Mu A- cell extracts for supporting Mu transposition in a cell-free reaction. PMID- 2981874 TI - Effects of adrenocorticotropin on 17 alpha-hydroxylase activity and cytochrome P 450(17 alpha) synthesis in bovine adrenocortical cells. AB - The effects of adrenocorticotropin (ACTH) on 17 alpha-hydroxylase activity and cytochrome P-450(17 alpha) synthesis have been studied utilizing bovine adrenocortical cells in primary monolayer culture. A 20-fold stimulation of the conversion of pregnenolone to 17 alpha-hydroxypregnenolone was observed in postmitochondrial supernatant fractions from cells treated with ACTH as compared to controls. This increase in 17 alpha-hydroxylase activity was found to be due to a change in the Vmax and not a change in the Km(app). By immunoisolation of newly synthesized protein from an RNA-directed cell-free translation system we found that the level of P-450(17 alpha) was many-fold greater when RNA from ACTH stimulated cells was used, as compared to RNA from control cells. A similar pattern was obtained when the rate of P-450(17 alpha) synthesis was analyzed by immunoisolation from radiolabeled cellular protein from ACTH-stimulated cells. When the total amount of P-450(17 alpha) was measured by immunoblotting we found that the levels of enzyme present correlated with the 17 alpha-hydroxylase activity. In addition, we found that these ACTH-mediated effects could be mimicked by treatment of cells with analogs of cyclic AMP. These results indicate that the activity of P-450(17 alpha) is regulated primarily by cyclic AMP mediated changes in synthesis, probably at the transcriptional level, which in turn has a profound effect on the pattern of steroid secretion. Thus, we believe cytochrome P-450(17 alpha) to be a key regulatory enzyme in the steroidogenic pathway. PMID- 2981875 TI - Activator protein required for the enzymatic hydrolysis of cerebroside sulfate. Deficiency in urine of patients affected with cerebroside sulfatase activator deficiency and identity with activators for the enzymatic hydrolysis of GM1 ganglioside and globotriaosylceramide. AB - Urine specimens from two sibs affected with cerebroside sulfatase activator deficiency were examined to ascertain whether the deficiency of the supplementary activator protein required for the enzymatic hydrolysis of cerebroside sulfate was also evident in urine. Material from chromatographic fractionations was examined for the activator activity to avoid ambiguities resulting from protein inhibition. There were substantial deficits in all chromatographic fractions corresponding to activator-containing fractions of control urines. Since patient urines contained elevated amounts of lactosylceramide, digalactosylceramide, and globotriaosylceramide and since similarities between activators for cerebroside sulfate and GM1 ganglioside hydrolyses had been noted previously, the chromatographic fractions were also examined for activators in other glycosphingolipid hydrolase systems. There was coincidence of activators for the GM1 ganglioside/beta-galactosidase and the globotriaosylceramide/alpha galactosidase A reactions with the cerebroside sulfatase activator in control urine fractions, and the patients' urines were deficient in activator activities for the three reactions. Identity of the three activators was suggested and antiserum to purified GM1 ganglioside activator was used to test this possibility. There were depressed levels of cross-reacting material in fractions of patient urines by Ouchterlony double diffusion and in unfractionated urine by enzyme-linked immunosorbent assay. Purified activators for the cerebroside sulfate and GM1 ganglioside systems showed lines of identity with no spurring on Ouchterlony double diffusion, identical mobility on immunoelectrophoresis, and similar stimulatory activities toward hydrolysis of the three glycosphingolipid species by their respective enzymes. Finally, the three activator activities were retained by anti-GM1-activator IgG coupled to Sepharose 4B. The results suggest strongly that the same protein entity serves as activator for the enzymatic hydrolysis of cerebroside sulfate, GM1 ganglioside, and globotriaosylceramide. PMID- 2981876 TI - The human receptor for T-cell growth factor. Evidence for variable post translational processing, phosphorylation, sulfation, and the ability of precursor forms of the receptor to bind T-cell growth factor. AB - The T-cell growth factor (TCGF) receptor on phytohemagglutinin-activated normal peripheral blood T-cells is characterized as a glycoprotein with an apparent Mr = 55,000 that contains N-linked and O-linked carbohydrate with only approximately 33,000 daltons of peptide structure (p33) as evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. There are two N-linked glycosylated intermediate precursor forms (apparent Mr = 35,000 (p35) and 37,000 (p37]. This receptor differs from the TCGF receptor on HUT-102B2 cells (apparent Mr = 50,000) because of differences in post-translational processing. Experiments with the carboxylic ionophore monensin demonstrate blockade of the transition of the p35 and p37 receptor precursor forms to the mature receptor, presumably secondary to inhibition of Golgi-associated receptor processing. We identify the primary translation product of TCGF receptor mRNA as intermediate in size between the p33 and the p35/p37 forms. We further demonstrate that the p33, p35, and p37 precursor forms, but not the primary translation product, are all capable of binding TCGF. Thus, the removal of the signal peptide and/or conformational changes of the primary translation product are necessary for ligand binding; however, the extensive post-translational modifications are not. Lastly, we demonstrate that at least some TCGF receptors are phosphorylated and sulfated, and that TCGF receptors on phytohemagglutinin-activated normal T-cells are more heavily sulfated than those on HUT-102B2 cells. PMID- 2981877 TI - Properties of the nerve growth factor receptor. Relationship between receptor structure and affinity. AB - Microsomal membranes from A875 human melanoma cells contain nerve growth factor receptors (NGF-receptors) which appear to belong to a single class with homogeneous binding properties, as determined by Scatchard plots. NGF-receptors in these membrane preparations are also uniformly highly sensitive to tryptic proteolysis, and 125I-NGF bound to NGF-receptor in these membranes is rapidly dissociated in the presence of a high concentration of unlabeled NGF. However, analysis of 125I-NGF dissociation kinetics indicated that two classes of NGF receptor were present in these membranes. Thus, NGF-receptors can express either high or low affinity trypsin-sensitive states in addition to the high affinity trypsin resistant NGF-receptor state described previously (Buxser, S. E., Kelleher, D. J., Watson, L., Puma, P., and Johnson, G. L. (1983) J. Biol. Chem. 258, 3741-3749). The high affinity trypsin-sensitive and low affinity trypsin sensitive states correlate with 200- and 90-kDa 125I-NGF X NGF-receptor complexes observed in photoaffinity cross-linking experiments. The absence of differences in peptide maps generated from the two sizes of NGF-receptor proteins together with structural and binding data strongly indicates that the 200-kDa NGF-receptor protein is a complex, probably a dimer, consisting of two 80-kDa NGF-receptor proteins associated with a single beta-NGF dimeric molecule. A model is proposed which relates structural states of NGF-receptors with specific receptor binding properties. The model provides an alternative explanation for binding phenomena previously attributed to negative cooperativity. PMID- 2981878 TI - HLA-D region antigen-associated invariant polypeptides as revealed by two dimensional gel analysis. Glycosylation and structural inter-relationships. AB - Two-dimensional polyacrylamide gel analyses of immunoprecipitates of HLA-D region antigens prepared from [35S]methionine-labeled B lymphoblastoid cells revealed a number of invariant polypeptides (Ii and theta) that co-precipitate with the alpha and beta polypeptides of the class II (Ia) antigens. The invariant polypeptides comprised at least three Ii spots of Mr = 31,000 (Ii1-Ii3) and a series of six theta spots of Mr = 34,000 (theta 1-theta 6). The structural inter relationships of these polypeptides have been investigated. Tryptic peptide fingerprints showed that Ii and theta have closely related amino acid sequences. In contrast, the fingerprints of the HLA-DR alpha and beta polypeptides clearly differed from those of theta and Ii as well as from each other. Analyses of immunoprecipitates prepared from cells cultured in the presence of tunicamycin revealed the presence of two N-linked oligosaccharides on each invariant polypeptide and suggested that the more acidic theta polypeptides (theta 1 and theta 2) differed from the other invariant polypeptides by the presence of sialic acid on one or both N-linked oligosaccharides. Removal of sialic acid by neuraminidase simplified the pattern of theta spots into three distinct Ii related polypeptides. Endo-beta-N-acetylglycosaminidase H digestion indicated that the individual theta polypeptides represent stages in carbohydrate processing whereby Ii with two N-linked immature oligosaccharides are converted initially to theta 6-theta 3 with one immature and one complex, but nonsialylated, oligosaccharide and finally to theta 2-theta 1 with two complex oligosaccharides. Digestion of the theta polypeptides with N-acetylgalactosamine oligosaccharidase indicated that the theta spots are also derived by O glycosylation from the Ii polypeptides. This assignment is supported by results obtained using monensin to block glycosylation within the Golgi. At least three spots persisted after complete removal of the N- and O-linked oligosaccharides, suggesting the presence of a family of invariant polypeptides differing in amino acid sequence. PMID- 2981880 TI - Characterization of an RNA polymerase activity from HeLa cell mitochondria, which initiates transcription at the heavy strand rRNA promoter and the light strand promoter in human mitochondrial DNA. AB - An RNA polymerase activity capable of initiating transcription at both the heavy strand rRNA promoter and the light strand promoter of human mitochondrial DNA has been partially purified from HeLa cell mitochondria and characterized in its requirements and products. The ratio of the two transcription initiating activities varied considerably from preparation to preparation. The human mtRNA polymerase partially purified by DEAE-cellulose and heparin-agarose chromatography exhibits a great sensitivity to ionic strength and to Mn2+, characteristics which clearly differentiate this enzyme from bacterial and eukaryotic nuclear RNA polymerases, and in contrast resemble the behavior of the yeast mtRNA polymerase. The human mtRNA polymerase exhibits a requirement for ATP which is 15- to 20-fold higher than that for the other NTPs, a low optimum template DNA concentration, and a marked susceptibility to inhibition by non mitochondrial DNA. PMID- 2981879 TI - Synthesis of an altered type III procollagen in a patient with type IV Ehlers Danlos syndrome. A structural change in the alpha 1(III) chain which makes the protein more susceptible to proteinases. AB - The synthesis of type III procollagen was examined in cultured fibroblasts from ten patients with type IV Ehlers-Danlos syndrome, a heritable disorder of connective tissue. With fibroblasts from nine patients, a decreased amount of labeled type III procollagen was recovered in the medium after the cells were incubated with radioactive amino acids for 24 h. The results were compatible with undefined defects in type III procollagen. The culture medium from one patient contained apparently normal amounts of type III procollagen after a 24-h labeling. However, the pro-alpha 1(III) chains from the medium of the patient's fibroblasts appeared as an abnormally broad band when examined by gel electrophoresis in sodium dodecyl sulfate. Analysis of fragments generated by vertebrate collagenase and cyanogen bromide located a structural defect between amino acid residues 555 and 775 in half of the alpha 1(III) chains. Most of the patient's type III procollagen was susceptible to digestion by pepsin or a mixture of chymotrypsin and trypsin at temperatures at which normal type III procollagen resisted digestion. Cyanogen bromide digestion of samples of the patient's skin revealed that the amount of type III was reduced more than 4-fold. The results support the hypothesis that both normal and structurally altered pro alpha 1(III) chains are being incorporated into type III procollagen synthesized by the patient's fibroblasts and that type III procollagen molecules containing one, two, or three structurally altered pro-alpha 1(III) chains are rapidly degraded by proteinases in the tissues. PMID- 2981881 TI - Stretch of the femoral nerve in a dancer. A case report. PMID- 2981882 TI - Virus-like filamentous intranuclear inclusions in a giant-cell tumor, not associated with Paget's disease of bone. A case report. PMID- 2981883 TI - Fibrosarcoma of long bones. A study of the significance of areas of malignant fibrous histiocytoma. AB - A study was made of the clinical course of 102 cases of fibrosarcoma of long bones. Of these, 55 were predominantly fibroblastic or pure fibrosarcomata while 47 contained extensive areas resembling so-called malignant fibrous histiocytoma (MFH). Treatment was similar in the two groups, and the five-year survival was the same, 34%, in both groups. Lung metastases developed in 63% of the fibroblastic sarcomata and 59% of the MFH-like tumours. Our study indicates that there is no significant difference in behaviour in fibrosarcoma with or without marked MFH features. The histological grading of both groups of fibrosarcomata together was of prognostic value; five-year survival was 64% in 14 Grade I tumours, 41% in 32 Grade II tumours and 23% in 56 Grade III tumours. PMID- 2981884 TI - Dissociation of the chemotactic and mitogenic activities of platelet-derived growth factor by human neutrophil elastase. AB - Because platelet-derived growth factor (PDGF) may be released at sites where neutrophil proteinases may also be released, we examined the effects of neutrophil elastase and cathepsin G upon the chemotactic and mitogenic activities of PDGF. Elastase abolished the chemotactic activity of PDGF for fibroblasts but had no effect on its chemotactic activity for monocytes, or on its mitogenic activity for 3T3 cells or its capacity to bind to 3T3 cells. Cathepsin G had no effect upon the chemotactic or mitogenic activities of PDGF. In contrast, trypsin eliminated the chemotactic activity of PDGF for monocytes and fibroblasts and the mitogenic activity of PDGF. After reduction and alkylation, PDGF retained full chemotactic activity for fibroblasts and monocytes but exhibited no mitogenic activity and only limited binding to 3T3 cells. These results indicate separate domains on PDGF for fibroblast chemotactic and mitogenic activity and for monocyte and fibroblast chemotactic activity and raise the possibility that the biological activities of PDGF may be modified selectively in vivo. The findings further suggest that the majority of PDGF receptors on fibroblasts mediate mitogenic activity and that only a minority of the PDGF receptors on fibroblasts are responsible for chemotactic activity. PMID- 2981885 TI - Site-specific maturation of enveloped viruses in L cells treated with cytochalasin B. AB - Treatment of infected L cells with 10 micrograms/ml cytochalasin B (CB) was found to promote a rapid relocalization of viral glycoproteins on the cell surface. Whereas the vesicular stomatitis virus G protein and the influenza virus hemagglutinin were uniformly distributed on the surface of untreated cells, in CB treated cells, they were strikingly concentrated at cell extremities in the regions of clustered blebs. Glycoprotein concentration at cell extremities was accompanied by preferential maturation of virus particles from the same sites; both vesicular stomatitis and influenza viruses budded predominantly from the vicinity of clustered blebs. This effect of CB was completely reversible. Removal of CB from the cell growth medium resulted in a return of viral glycoproteins to the uniform distribution characteristic of untreated cells and to uniform virus budding. The results of this study are interpreted in terms of a model that suggests that preferential budding of viruses from the regions of bleb clusters is due to the concentration of viral glycoproteins at these sites. PMID- 2981886 TI - Highly specific antibody to Rous sarcoma virus src gene product recognizes a novel population of pp60v-src and pp60c-src molecules. AB - Antiserum to the Rous sarcoma virus (RSV)-transforming protein, pp60v-src, was produced in rabbits immunized with p60 expressed in Escherichia coli. alpha p60 serum immunoprecipitated quantitatively more pp60v-src than did tumor-bearing rabbit (TBR) sera. When RSV-transformed cell lysates were preadsorbed with TBR serum, the remaining lysate contained additional pp60v-src, which was recognized only by reimmunoprecipitation with alpha p60 serum and not by TBR serum. In subcellular fractions of RSV-infected chicken embryo fibroblasts (RSV-CEFs) and field vole cells probed with TBR serum, the majority of the pp60v-src was associated with the plasma membrane-enriched P100 fraction. However, alpha p60 serum revealed equal distribution of pp60v-src and its kinase activity between the P1 (nuclear) and P100 fractions. The same results were obtained for pp60c-src in uninfected CEFs. On discontinuous sucrose gradients nearly 50% of the P1-pp60v src sedimented with nuclei, in fractions where no plasma membrane was detected. Indirect immunofluorescence microscopy of RSV-CEFs with alpha p60 serum revealed a distinct pattern of perinuclear fluorescence, in addition to staining at the cell periphery. Thus the use of a highly specific antibody reveals that enzymatically active pp60v-src and pp60c-src molecules are present in other intracellular structures, probably juxtareticular nuclear membranes, in addition to the plasma membrane in normal, uninfected, and wild-type RSV-infected cells. PMID- 2981887 TI - Mobility of fluorescent derivatives of cytochrome c in mitochondria. AB - Motion of cytochrome c bound to giant (2-10-micron diam) mitochondria isolated from the waterbug Lethocerus indicus was examined using the technique of fluorescence recovery after photobleaching. Fluorescent cytochrome c was exchanged for native cytochrome c through partly damaged outer membrane. Recovery profiles were not statistically different when the fluorescence from iron-free cytochrome c or fluorescein-labeled cytochrome c was used and were essentially the same in the presence or absence of an uncoupler. In the presence of excess porphyrin cytochrome c, the apparent diffusion coefficient was 6 X 10(-11) cm2/s in 0.3 M sucrose-mannitol-EDTA and 3 X 10(-10) cm2/s in 0.10 M KCl/0.10 M sucrose. At concentrations of porphyrin cytochrome c that are stoichiometric with cytochrome c oxidase and for mitochondria in which excess cytochrome c was washed away, two components were observed in the recovery profile. The diffusion coefficient of the fast component was 1 X 10(-10) cm2/s. The second component showed no recovery during the time scale of measurement (D less than 10(-12) cm2/s). We speculate on the origin of the immobile fraction. PMID- 2981888 TI - Maize plastid photogenes: mapping and photoregulation of transcript levels during light-induced development. AB - Positively photoregulated regions that show increased transcript levels upon illumination of dark-grown seedlings are scattered over approximately 19% of the maize plastid chromosome. Some photogenes, i.e., genes within these regions, are transcribed individually, whereas others that are transcribed as polycistronic mRNAs appear to be functionally organized into operons. Multiple light-induced transcripts are complementary to most photogenes; these mRNAs are not present in equimolar amounts during plastid photomorphogenesis, but particular transcripts predominate at specific stages of development. Most, but not all, photogene RNA pools reach a maximum size (after either 10, 20, or 44 h of illumination) and then fall to approximately preillumination levels. These data and other considerations argue that photogene expression control is fundamentally transcriptional and that there is more than one expression class. Transcripts of the maize plastid gene for the large subunit of ribulose bisphosphate carboxylase reach a maximum by 20 h of illumination; transcripts of the nuclear gene for the small subunit of this enzyme continue to accumulate and fall considerably later. These data suggest that the level of transcription of the latter gene in the nucleus may be regulated by events in the chloroplast. PMID- 2981890 TI - Translatable mRNA for GP140 (a subunit of type VI collagen) is absent in SV40 transformed fibroblasts. AB - Production of GP140, a major component of the extracellular matrix of cultured fibroblasts, is markedly decreased in SV40 transformed cells as compared with normal cells (Carter, W. G., 1982, J. Biol. Chem., 257:13805-13815). To determine at what step the biosynthesis is inhibited, we compared the levels of functional mRNA for GP140 in normal and transformed fibroblasts. Translation of total RNA from W138 cells in a reticulocyte lysate, followed by immunoprecipitation with affinity-purified antibodies to GP140, yielded a single polypeptide with an Mr of 125,000. This polypeptide was identified as GP140 based on its immunoreactivity, collagenase sensitivity, and comigration on polyacrylamide gels with GP140 synthesized by cells in the presence of tunicamycin and 2,2'-bipyridyl. No cell free synthesis of GP140 was observed with total RNA from SV40 transformed W138 cells, indicating that these cells contain very low levels of GP140-specific mRNA. The biosynthesis of GP140 might therefore be blocked at the transcriptional level. PMID- 2981889 TI - Novel peripheral blood-derived human cell lines with properties of megakaryocytes. AB - For 18 mo, we derived 18 cell lines from 11 donors with various clinical profiles ranging from normal to leukemic. Suspension cultures were initiated with 1 X 10(6) mononuclear blood cells/ml of nutrient medium containing 10% human serum and 10% lectin-stimulated human lymphocyte conditioned medium. The cultures were monitored weekly by morphological analyses of Wright-Giemsa-stained cell preparations. All successful cultures showed a significant decline in viability during the first 3-4 wk with rate "lymphoid" cells observed in mitosis. Within the next 2 wk, the proliferating cells gave rise to a rapidly expanding population of mononuclear cells. As the cultures expanded, cell morphology became heterogeneous with respect to cell size and nuclear ploidy, with an accumulation of giant multinuclear cells that were suggestive of megakarocytes. Even though the cells did not have the classical morphology of mature platelet-forming megakaryocytes, 90% of the cells within a cell line were positive by direct or indirect immunofluorescence for the platelet membrane glycoproteins IIb and IIIa; for surface markers HLA-Dr and B2-microglobulin; for intracellular platelet derived growth factor and platelet factor IV; and for membrane affinity or binding with serum platelet-derived growth factor and platelet factor IV. These results suggest that a blood precursor cell, most likely a primitive megakaryoblast, was isolated from the peripheral blood and was provided with an optimal culture environment for sustained growth. These cells did not mature to a more differentiated stage, perhaps owing to regulatory factor deficiencies in this in vitro system. The remarkable frequency of obtaining cell lines with megakaryocyte properties from normal peripheral blood and the capacity of some normal donors to repeatedly yield these cell lines make this cell culture system indeed unique by being selective for putative megakaryocyte precursors. PMID- 2981891 TI - Neutrophil degranulation: evidence pertaining to its mediation by the combined effects of leukotriene B4, platelet-activating factor, and 5-HETE. AB - Stimulus-activated polymorphonuclear neutrophils (PMN) produce leukotriene B4 (LTB4), 5-hydroxyeicosatetraenoate (5-HETE), and platelet-activating factor (PAF). Each of these lipids promotes PMN degranulation; in combination they have additive and potentiating effects that result in prominent degranulation responses at relatively low concentrations. Thus, the combined interactions of LTB4, 5-HETE, and PAF may mediate responses in PMN activated by other stimuli. This possibility was examined by measuring the responses of PMN made insensitive to one or more of these lipids. Cells were pretreated with LTB4, 5-HETE, and/or PAF for 8 min; exposed for 2 min to cytochalasin B (which is required for lipid induced degranulation); and then challenged. PMN challenged with only buffer released minimal amounts of granule-bound enzymes. Furthermore, the lipid pretreated cells were hyporesponsive to challenge with 1) various combinations of these same lipids or 2) ionophore A23187. The relative potencies of the lipids in producing hyporesponsiveness to themselves or A23187 were: 5-HETE less than PAF less than or equal to LTB4 less than PAF + LTB4 less than PAF + LTB4 + 5-HETE. For both types of challenge, reduced responsiveness occurred in cells pretreated with greater than 0.1 nM LTB4 and/or greater than 0.2 nM PAF, persisted in cells washed after lipid pretreatment, and did not develop in cells pretreated with various combinations of bioinactive structural analogues of the lipids. Thus, PAF, LTB4, and 5-HETE interacted to desensitize PMN, and the degranulating actions of A23187 required cells that were fully responsive to each of the three lipids. This supports the concept that the lipids act together in mediating certain of the ionophore's effects. However, lipid-desensitized PMN degranulated fully when challenged with C5a, a formylated oligopeptide, or phorbol myristate acetate. Degranulation responses, therefore, may proceed through various pathways, only some of which involve the lipid products studied here. PMID- 2981892 TI - Limb development in chick embryos: cyclic AMP-dependent protein kinase activity, cyclic AMP, and prostaglandin concentrations during cytodifferentiation and morphogenesis. AB - The effects of prostaglandin E2 (PGE2) on cyclic AMP (cAMP) concentrations of chick limb bud cells obtained from limbs at various stages of development were investigated. In addition, endogenous concentrations of PGE2 were examined in whole limbs from comparable stages. Prior to either chondrogenesis or myogenesis (stages 20-23), cells were more responsive to PGE2, in terms of cAMP levels, than those of differentiated phenotypes, obtained at stages 25-28. This greater responsiveness to PGE2 of undifferentiated cells was correlated with endogenous concentrations of PGE2 which were significantly higher in undifferentiated limbs than in limbs containing differentiated cartilage and muscle. Cyclic AMP dependent protein kinase (PKA) activity was detectable in cell homogenates at each stage examined and did not appear to change in cAMP dependency at any stage. The majority (80-85%) of total enzyme activity was localized in soluble fractions of cell homogenates while the residual activity was localized to membrane enriched, particulate fractions. The results demonstrate that both responsiveness of limb mesenchyme to PGE2 and endogenous concentrations of PGE2 are maximal prior to cytodifferentiation of limb tissues. The presence of cAMP-dependent protein kinase in these undifferentiated cells supports a regulatory role for both PGE2 and a cAMP-protein phosphorylation system in the differentiation of limb tissues. PMID- 2981893 TI - Intracellular calcium redistribution and its relationship to fMet-Leu-Phe, leukotriene B4, and phorbol ester induced rabbit neutrophil degranulation. AB - The addition of low concentrations (less than 10(-7) M) of the calcium ionophore A23187 to rabbit neutrophils releases the intracellular pool of calcium previously shown in radioactive steady-state and chlortetracycline fluorescence studies to be mobilized by chemotactic factors. A23187 at these concentrations elicits no functional responses from these cells. However, A23187, added before chemotactic factors such as fMet-Leu-Phe and leukotriene B4, inhibits the ability of the latter stimuli to induce, in the presence of cytochalasin B, an exocytotic release of the neutrophil's cytoplasmic granules. These results imply that the chemotactic-factor-induced release of intracellular calcium is a necessary event for the optimal activation of the neutrophils. Phorbol ester-induced neutrophil degranulation on the other hand is unaffected by exposure to A23187, thereby completely dissociating its mechanism of action from rises in cytoplasmic free calcium. PMID- 2981894 TI - Characterization of chemically and virally transformed variants of Madin-Darby canine kidney (MDCK) epithelial cells. AB - Oncogenic derivatives of Madin-Darby canine kidney (MDCK) cells were isolated in the nude mouse, and nononcogenic anchorage-independent transformants were isolated in vitro following chemical mutagenesis in vitro. These transformed cell lines as well as a Moloney sarcoma virus (MSV) transformed line were characterized with respect to their serum and anchorage requirements, growth rates, final saturation densities, and sensitivities to contact inhibition. None of these in vitro growth characteristics were found to correlate with tumorigenicity in nude mice. One tumorigenic clone, MDCK-T1, was characterized with respect to serum-free growth requirements, cAMP production, and ornithine decarboxylase (ODC) activity. These cells exhibited a significant reduction in the PGE1 requirement for growth, they produced higher levels of cAMP, and they expressed a reduced level of ODC activity relative to the parental MDCK cells. These findings may reflect changes in growth control mechanisms which accompany kidney epithelial cell tumorigenesis and suggest that the study of transformed lines derived in this manner could lead to the identification of in vitro properties which are associated with malignancy. PMID- 2981895 TI - Lineage specific receptors used to identify a growth factor for developmentally early hemopoietic cells: assay of hemopoietin-2. AB - A new approach, based on the occurrence of receptors for the mononuclear phagocyte lineage specific hemopoietic growth factor (HGF) colony stimulating factor-1 (CSF-1) on developmentally early multipotent cells, is utilized to detect and assay rapidly another HGF, hemopoietin-2. This method is also used to determine the relative maturity of hemopoietin-2 target cells, to investigate synergism between hemopoietin-2 and CSF-1, and to measure CSF-1 receptor levels on maturing cells. While the target cell specificities of hemopoietin-2 and CSF-1 overlap, hemopoietin-2 causes the appearance of developmentally earlier 125I-CSF 1 binding cells de novo in the absence of CSF-1. Increased CSF-1 receptor densities are observed on cells incubated with either HGF, consistent with acquisition of the capacity for increased expression of the receptor by mononuclear phagocyte progenitor cells just prior to their differentiation to adherent mononuclear phagocytes. Together, both HGFs have a synergistic effect on the generation of 125I-CSF-1 binding cells with elevated CSF-1 receptor densities. Preliminary characterization of hemopoietin-2 from medium conditioned by WEHI-3 cells indicates that it is very similar to, if not identical with, interleukin-3 (IL-3) and the HGF(s) acting on multipotential cells and cells giving rise to erythroid cells, granulocytes, mononuclear phagocytes, and megakaryocytes. Purified IL-3 was shown to possess hemopoietin-2 activity. PMID- 2981896 TI - Synergism between hemopoietic growth factors (HGFs) detected by their effects on cells bearing receptors for a lineage specific HGF: assay of hemopoietin-1. AB - The preceding paper describes a new approach to the detection and assay of growth factors for developmentally early multipotent hemopoietic cells (Bartelmez et al., J. Cell. Physiol., 1985). This approach, involving measurement of the increase in the number of receptors for the mononuclear phagocyte specific hemopoietic growth factor (HGF), colony stimulating factor-1 (CSF-1), in cultures of developmentally early murine cells incubated with putative HGFs, has been used to define and assay hemopoietin-1. Hemopoietin-1 (Mr approximately 20,000) is found in the medium derived from serum-free cultures of cells of the human urinary bladder carcinoma line 5637. In contrast to both hemopoietin-2 and CSF-1, which also stimulate an increase in CSF-1 receptor numbers in cultures of developmentally early hemopoietic cells, hemopoietin-1 alone has no detectable effect. However, hemopoietin-1 exhibits dramatic synergism with CSF-1. In the presence of CSF-1, hemopoietin-1 stimulates the proliferation of developmentally earlier cells than those that respond to either CSF-1 alone or hemopoietin-2 alone or their combination. These cells proliferate for at least 3 days with no alteration of the average CSF-1 receptor density. However, by 5 days of incubation, the progeny of developmentally early hemopoietic cells that have proliferated in response to hemopoietin-1 + CSF-1 exhibit an approximately tenfold increase in the average CSF-1 receptor density per cell, which immediately precedes their differentiation to adherent mononuclear phagocytes. As hemopoietin-1 does not possess colony stimulating or burst promoting activities for murine bone marrow cells, but acts on multipotent hemopoietic cells, the analysis of the mechanism of its synergistic effects with HGFs such as CSF-1 are of special relevance to the regulation of early events in hemopoiesis. PMID- 2981897 TI - Uridine phosphorylase from Novikoff rat hepatoma cells: purification, kinetic properties, and its role in uracil anabolism. AB - Uridine phosphorylase activity was detected in sonic extracts of six different mammalian cell lines and, in conjunction with uridine kinase, provides a route for the conversion of uracil to UMP via uridine. Uracil phosphoribosyl transferase activity was not detected in any of eight different mammalian cell lines. Uridine phosphorylase was purified 5,330-fold from Novikoff rat hepatoma cells by ammonium sulfate precipitation, DEAE-Sephadex chromatography, hydroxyapatite chromatography, and Sephadex G-200 fractionation. The molecular weight of the enzyme by gel filtration was approximately 45,000. The kinetics of the purified enzyme were analyzed with respect to all four substrates at saturating cosubstrate concentration, yielding the parameters KmUra = 360 microM, KmRib-1-P = 88 microM, KmUrd = 16 micron, and KmPi = 130 microM. However, in intact cells the phosphorolysis of uridine proceeded with an apparent Km of 231 microM. Novikoff cells treated with 0.5 mM inosine exhibited an increase in uracil uptake rate which was proportional to an observed increase in intracellular ribose-1-phosphate. Nevertheless, in cells whose de novo synthesis of pyrimidines was blocked by pyrazofurin or N-(phosphonacetyl)-L-aspartate ("PALA"), the uptake of uracil was insufficient to support proliferation, even when enhanced by inosine. These observations are consistent with the kinetic characteristics of the enzyme and provide evidence that the intracellular level of ribose-1-phosphate plays a rate-limiting role in the uptake of uracil mediated by uridine phosphorylase. PMID- 2981898 TI - Regulation of K562 cell transferrin receptors by exogenous iron. AB - Single-cell analysis of K562 human erythroleukemia cells by flow cytometry was used to demonstrate the specific role of iron in regulating transferrin receptors (TfRs) and to establish that TfR expression does not necessarily correlate with growth rate. Exogenous iron concentration in culture was manipulated by supplementing the medium with sera having different iron concentrations over the range 0.6 to 5.4 micrograms/ml, by the addition of iron in the form of FeCl3, iron-saturated serum, or diferric transferrin, and by the addition of the iron chelator Desferal (desferrioxamine). TfR expression was negatively correlated with exogenous iron content: any treatment that reduced exogenous iron supply by at least 15% resulted in as much as a 1.8-fold increase in external receptors, detected as binding by both transferrin and monoclonal anti-TfR antibodies, and a 1.5-fold increase in the pool of internal receptors, as detected by anti-TfR antibody binding. None of these treatments altered growth rate, total cellular protein content, protein synthetic rate, cell cycle distribution or cell size. The rapid (12 hr) and reversible induction of internal and external receptors by Desferal was inhibited by cycloheximide and therefore may have resulted from de novo synthesis and not just mobilization of internal receptor pool to the cell surface. The correlation between growth rate and TfR expression previously observed in these and other cells must be secondary to cellular mechanisms that maintain intracellular iron pools by regulating synthesis, recycling, and cell surface expression of TfRs. PMID- 2981899 TI - Isolation and characterization of six new genome types of human adenovirus types 1 and 2. AB - Several of the 41 types of human adenovirus have been divided into genome types based on aberrant restriction endonuclease digestion patterns of viral DNA. In the process of screening a large number of clinical adenovirus isolates by restriction endonuclease digestion of viral DNA, we have identified nine isolates from eight patients which were identified by neutralization as adenovirus type 1 or 2 but had aberrant cleavage patterns. Cleavage sites for the enzymes SmaI, EcoRI, HindIII, KpnI, and HpaI were mapped. The variants could be placed in six distinct groups based on cleavage patterns. The designation genome types 1a, 1b, 1c, 2b, 2c, and 2d are proposed for these isolates. PMID- 2981900 TI - Bacterial chromosomal restriction endonuclease analysis of the homology of Bacteroides species. AB - Chromosomal DNAs of selected Bacteroides organisms whose relatedness had been previously determined by "conventional" filter-annealing studies (J. L. Johnson, Int. J. Syst. Bacteriol. 28:245, 1978) were further analyzed by restriction endonuclease analysis coupled with the Southern hybridization procedure (E. M. Southern, J. Mol. Biol. 98:503, 1975). By comparing their EcoRI restriction fragment patterns in agarose gel electrophoresis, each Bacteroides strain could be clearly differentiated. As a simple and direct means for comparison purposes this method was particularly useful for differentiating genetically similar organisms such as Bacteroides strains of the same species which shared greater than 75% homology. In contrast, bacterial chromosomal restriction endonuclease analysis in conjunction with Southern hybridizations was most effectively used to determine the significance of low levels of homology (less than 24%) as this technique provided additional information on the nature and relative distribution of that homology when the areas of homology were displayed as reproducible bands in autoradiograms. PMID- 2981901 TI - Detection and serotyping of herpes simplex virus in MRC-5 cells by use of centrifugation and monoclonal antibodies 16 h postinoculation. AB - Monoclonal antibodies (Syva Co., Palo Alto, Calif.) were used for the detection and serotyping of herpes simplex virus (HSV) isolates by immunofluorescence 16 h after inoculation of MRC-5 monolayers in 3.7-ml shell vials and after low-speed centrifugation. A total of 119 specimens were inoculated into conventional tube cell cultures and shell vials. Of 98 specimens inoculated on the same day of receipt in the laboratory (fresh specimens), all 23 (23.5%) HSV-positive specimens were identified by serotype in 16 h in shell vials by immunofluorescence, whereas only 8 of 23 HSV-positive specimens (34.8%) produced cytopathic effects in conventional tube cell cultures in this time period. Similarly, of 21 original specimen extracts previously determined to be culture positive for HSV (stored specimens), all were detected and serotyped by the immunofluorescence test with monoclonal antibodies 16 h postinoculation compared with the recognition of only 8 of these isolates (38.1%) by cytopathic effect that soon. This technique of centrifugal inoculation of HSV in shell vials containing MRC-5 cells permitted detection of this virus in all positive specimens with serotype determination within 16 h postinoculation. PMID- 2981902 TI - Comparison of enzyme-linked immunosorbent assay with enzyme-linked fluorescence assay with automated readers for detection of rubella virus antibody and herpes simplex virus. AB - The enzyme-linked immunosorbent assay (ELISA) was compared with the enzyme-linked fluorescence assay (ELFA) for the detection of rubella antibody and herpes simplex virus antigen. Test parameters, specimens, antigen or antibody, and conjugates for the two types of assays were identical except that p-nitrophenyl phosphate was used as the substrate for the ELISA and 4-methylumbelliferyl phosphate was used as the substrate for ELFA. Automated readers were used for both assays. Antibody titers and sensitivity of antigen detection were quite similar for ELISA and ELFA. ELFA for rubella antibody, however, could be conducted with less antigen or shorter substrate incubation time (5 min for ELFA versus 30 min for ELISA). For herpes simplex virus antigen detection, ELFA could also be read after a shorter substrate incubation time (15 min for ELFA versus 30 min for ELISA). Clear polystyrene microtiter plates routinely used for ELISA could be used for ELFA, but clear polyvinyl chloride plates had high background fluorescence. Black polystyrene and polyvinyl chloride plates gave lower background fluorescence than did clear plates. ELFA is of particular value as a substitute for ELISAs in which long substrate incubations are required or antigens of only low titer are available. PMID- 2981903 TI - Micromodified cytomegalovirus antibody screening test. PMID- 2981905 TI - Heart transplant pathology: the British experience. AB - An account of human heart transplantation as seen by the histopathologists involved at the two UK transplant centres is presented. Between January 1979 and July 1984 179 patients received 186 hearts and 124 are still alive up to four years after operation. Cyclosporin A based immunosuppression has been used in the last 120 patients. Four patients developed neoplastic lesions. The commonest reason for transplantation was ischaemic heart disease (63%), followed by congestive cardiomyopathy (35%). The seven retransplants were for acute or chronic rejection. The monitoring of rejection by endomyocardial biopsies is described, and the causes of death and necropsy findings are presented. PMID- 2981906 TI - Anatomy of brain alpha 1-adrenergic receptors: in vitro autoradiography with [125I]-heat. AB - Much useful information on the localization of alpha 1-adrenergic binding sites has been gained by using tritiated radioligands for in vitro autoradiography. However, the iodinated alpha 1-adrenergic antagonist HEAT [( 2-beta (4 hydroxyphenyl)-ethylaminomethyl)-tetralone], BE 2254), a radioligand with high affinity and specificity, provides autoradiographs with a higher signal to noise ratio. This has allowed us to describe the anatomy of these binding sites in much greater detail than previously possible. Regions showing the highest levels of binding include external plexiform layer of the olfactory bulb, layers Va and Vc of frontoparietal cortex, lateral and central amygdaloid nuclei, thalamus, and inferior olive. Other regions were generally less intensely labeled, with the least evidence of labeling in white matter, such as corpus callosum. Some regions (e.g., hippocampus) had only moderate labeling, but the binding appeared in a discrete pattern that reflected the functional organization of the structure. Although the [125I]-HEAT binding sites were distributed in a pattern similar to that previously reported for [3H]-WB 4101 and [3H]-prazosin, the anatomical detail seen with the iodinated ligand is greater. As a result, an association of alpha 1-adrenergic antagonist binding sites with specific layers in the cortex and with some catecholamine-containing nuclei in the brainstem, such as the locus coeruleus, have been seen for the first time. PMID- 2981904 TI - Biochemical enzyme analysis in acute leukaemia. AB - This report summarises the current knowledge regarding the clinical utility of biochemical enzyme markers for both diagnostic and therapeutic purposes in acute leukaemia. The enzymes studied most extensively in this field are terminal deoxynucleotidyl transferase, adenosine deaminase, 5'-nucleotidase, purine nucleoside phosphorylase, and acid phosphatase, esterase, hexosaminidase isoenzymes. For each enzyme, the quantitative and qualitative characteristics in various immunologically defined subclasses of acute leukaemia are described. The quantitative evaluation of enzyme activities represents an adjunctive classification technique which should be incorporated into the multivariate analysis, the "multiple marker analysis." By qualitative characterisation pronounced heterogeneity of leukaemia subsets is uncovered. The application of 2' deoxycoformycin, a specific inhibitor of adenosine deaminase, and the potential usefulness of two other enzymes as targets for treatment with selective agents is discussed. The concept that gene products expressed at certain developmental stages of normal cells can similarly be detected in leukaemic cells (which therefore seem to be "frozen" or "arrested" at this particular maturation/differentiation stage) is supported by the results obtained in enzyme studies. Besides their practical clinical importance for classification and treatment of acute leukaemias, biochemical enzyme markers constitute a valuable research tool to disclose biological properties of leukaemic cells. PMID- 2981907 TI - Variability in the terminations of GABAergic chandelier cell axons on initial segments of pyramidal cell axons in the monkey sensory-motor cortex. AB - Chandelier cell axons were studied in the sensory-motor cortex of adult monkeys. The axonal fields of Golgi-impregnated chandelier cells in layer II in motor cortex are flattened sagittally. The vertical terminal portions of the axons varied both in length and in the numbers converging to form terminations of greater or lesser complexity. Golgi-impregnated plexuses were embedded in plastic and resectioned serially at 2.5-3.0 micrograms. A single axonal field could have as many as 400 terminal rows. All lie 3-13 micrograms beneath pyramidal cell somata. These terminations are not randomly distributed but instead, form clusters. Further resectioning the plastic sections for electron microscopy revealed that all the terminations are on the initial axon segments of pyramidal cells and all form symmetric synaptic contacts. In immunocytochemical material stained for glutamic acid decarboxylase (GAD), the enzyme involved in the synthesis of GABA, GAD-positive boutons were found to form symmetric synaptic contacts with a variety of postsynaptic elements including the axon hillocks and axon initial segments of pyramidal cells. Serial reconstructions from electron micrographs revealed GAD-positive terminals synapsing with the axon initial segment of pyramidal cells joined by cytoplasmic bridges and forming vertically oriented rows identical to those of chandelier cell terminals identified positively in the resectioned Golgi material. The GAD-positive terminals forming initial segment synapses were never continuous with GAD-positive terminals forming axo hillock synapses. The latter probably arise from basket cell axons. Initial segments of pyramidal cell axons in layers II and III were contacted by more GAD-positive terminals than the initial segments of pyramidal cell axons in layer V. The largest pyramidal cells in layer III received the most synapses. Many larger pyramidal cells, identified as callosally projecting cells by the retrograde transport of horseradish peroxidase (HRP), were shown in serial electron micrographs to possess large numbers of initial segment synapses, comparable to those seen in the immunocytochemical material. Serial reconstructions of pyramidal cell axons from axon hillock to the first myelin internode in resectioned Golgi, immunocytochemical and HRP material showed that the number of synapses varied from 2 to 52 for layers II and III and from 2 to 26 for layer V. The number of synapses on the axon hillocks varied from zero to 12. The variability in these terminations may be an important factor in the shaping of the functional properties of the pyramidal cells. PMID- 2981908 TI - Metastatic disease to the cavernous sinus: clinical syndrome and CT diagnosis. AB - We analyzed the clinical constellation of signs and symptoms and the radiographic studies of 17 patients with histologic verification of cavernous sinus metastases. Although most patients presented with acute, unilateral, painful ophthalmoplegia, and with a rapidly progressive course, the clinical diagnosis of metastatic disease was often delayed. This was probably due to the fact that, in the majority of patients, cavernous sinus symptoms were either the first expression of an unknown malignancy or the first manifestation of metastatic disease in those with a known primary. Computed tomography was found to be an indispensable diagnostic aid. In 16 of the 17 patients CT established the presence of an enhancing mass in the cavernous sinus sometimes associated with bone erosion. Thin section, contrast enhanced high resolution CT in axial and coronal projections represents the imaging procedure of choice for metastatic disease to the cavernous sinus. PMID- 2981909 TI - CT evaluation of glucagonomas. AB - Seven patients with clinical features of the glucagonoma syndrome, including the characteristic rash, diabetes mellitus, and weight loss, were examined by CT. Computed tomography demonstrated a primary pancreatic tumor in all patients, and, in four, hepatic metastases were identified. The primary tumors, relatively large and solid in nature, varied in size from 2.5 to 6 cm in maximum diameter. The tumor was found in the tail of the pancreas in three patients and in the head of the pancreas in four. No obliteration of adjacent perivascular or peripancreatic fat planes was observed in any patient. Calcification was present in the primary tumor in three patients and in the hepatic metastases in one. All tumors that were studied angiographically were found to be hypervascular. In contrast to insulinomas, which are frequently quite small when clinically diagnosed, glucagonomas appear to attain considerable size prior to being clinically apparent. Thus, we conclude that CT ought to become the mainstay in the identification, localization, and staging of these tumors. PMID- 2981911 TI - Hemangiolymphangioma of the tongue treated by transfixion technique. PMID- 2981910 TI - Evaluation of 2-phosphono-butane 1,2,4 tricarboxylate as a crystal growth inhibitor in vitro and in vivo. AB - The inhibitory activity of 2-phosphono-butane 1,2,4 tricarboxylic acid (PBTA) in the formation of hydroxyapatite (HA) was studied in vitro and in vivo. PBTA, at 4 ppm, inhibited the spontaneous formation of HA in vitro from a supersaturated solution. PBTA, at 2 ppm, completely inhibited the crystal growth of HA; at lower concentrations, a direct relationship was found between the reduction of the initial precipitation rates and PBTA concentrations in the solution. The effects of PBTA on human dental enamel in vitro were also evaluated at pH 5.0 and 7.5. Equimolar levels of EDTA served as the controls. In comparison with EDTA, the dissolution of enamel induced by PBTA was negligible. One percent solution of PBTA was evaluated in a rat calculus assay. In comparison with a placebo solution, it significantly (p = 0.05) reduced calculus formation when applied topically. It was also tested against calculus formation in beagle dogs. A topical application once a day of a 1% solution at pH 7.0 reduced calculus formation by 84% for 16 weeks. Analysis of these data suggests that the agent effectively reduces calculus formation in vivo. PMID- 2981913 TI - Glomus tumor of the metacarpophalangeal joint: a case report. AB - A case of a glomus tumor of the head of the third metacarpal bone is described. The metacarpophalangeal joint had to be opened to allow excision of the tumor. PMID- 2981912 TI - Adenosine-induced bronchoconstriction in asthma: role of mast cell-mediator release. AB - Adenosine, when it is administered by inhalation to asthmatic subjects, is a potent bronchoconstrictor, although its mechanism of action is not known. Since adenosine has been demonstrated to potentiate IgE-dependent mediator release from mast cells, we have investigated the possible relationship between adenosine induced bronchoconstriction and release of mast cell mediators in 14 asthmatic subjects. In the first study the effect of the putative mast cell-stabilizing drug cromolyn sodium (SCG) was observed on the dose-related changes in SGaw and FEV1 produced by inhaled adenosine and histamine in seven subjects. Inhaled SCG (20 mg) had no effect on the airway responses to histamine. In contrast SCG significantly protected against adenosine-induced bronchoconstriction in four of the seven subjects as reflected by a decrease in the airway response to the highest concentrations of adenosine, from 65 +/- 8% to 12 +/- 3% (mean +/- SEM) for SGaw and 31 +/- 7% to 8 +/- 3% for FEV1. Those three subjects whose adenosine response was unaffected by SCG had received regular SCG until 12 hr before the studies. In a separate study on eight subjects, a single inhalation of adenosine, causing a maximum 61 +/- 4% fall in SGaw at 10 min, had no significant effect on circulating levels of histamine, neutrophil chemotactic factor, or cyclic AMP. Together these two studies suggest that bronchoconstriction produced by adenosine is not a consequence of enhanced mast cell-mediator release and that the inhibitory effects of SCG occur by a mechanism other than through mast cell stabilization. PMID- 2981914 TI - Upper extremity neuropathies after cardiac surgery. AB - Although coronary artery bypass grafting is a common procedure, there can be significant postoperative morbidity. The occasional finding of unexplained postoperative neuropathy in these patients prompted this study. Fifty-three patients who had cardiac surgery that used the standard median sternotomy were studied prospectively. Detailed sensory and motor testing and intraoperative measurement of the distance of sternal retraction and duration of cardiopulmonary bypass were recorded. Twenty patients (37.7%) exhibited postoperative motor and sensory neuropathies, all of which involved the ulnar nerve. Five patients who were studied with electromyography and nerve conduction evaluations exhibited evidence of brachial plexus injury. The average duration of symptoms was 2.3 months, but several patients have long-term unresolved symptoms. Previous neuropathies, wide retraction of the sternum, and long cardiopulmonary pump runs seem to predispose to such injury, which appears to involve the brachial plexus. Anatomic reasons for such findings are offered. PMID- 2981915 TI - Transfer of antigen-presenting capacity to Ia-negative cells upon fusion with Ia bearing liposomes. AB - The role of Ia in T cell activation was investigated by incorporating affinity purified I-Ad molecules into synthetic liposomal membranes and by using these as antigen-presenting units. IL 2 production by I-Ad-restricted, chicken ovalbumin specific T cell hybridomas was measured in a system in which antigen processing by the presenter was not required. I-Ad-bearing liposomes were found to have no antigen-presenting capacity. It was shown, however, that antigen-presenting capacity could be conferred on Ia-negative cells by fusion of these cells with liposomes bearing I-Ad molecules, together with Sendai virus envelope glycoproteins, as fusogenic agents. Both Ia-negative B lymphoma cells and mouse L cells were capable of antigen presentation of predigested ovalbumin after fusion with vesicles formed from phosphatidylserine and phosphatidylethanolamine in a 1:1 w:w ratio. The cell surface expression of the transferred Ia remained stable for at least 7 hr. These results indicate that Ia is the only additional cell surface molecule required, at least by Ia-negative B cell lymphomas and L cells, to convert them into effective antigen-presenting cells. This system should be useful in future studies of the cellular requirements for antigen processing and presentation. PMID- 2981916 TI - Inhibition of mixed lymphocyte response by a rat monoclonal antibody to a novel murine lymphocyte activation antigen (MALA-2). AB - A novel surface antigen expressed on activated and proliferating murine lymphocytes has been identified by a rat monoclonal antibody. The antigen, termed MALA-2, is also expressed on various lymphoid cell lines, but it is absent or present at very low densities on the majorities of unstimulated thymocytes and lymph node cells. Some cells in normal spleen and bone marrow seem to express the antigen at relatively high densities and they may represent proliferating cells in these tissues. MALA-2 has an apparent m.w. of 95,000 to 100,000 under both reducing and nonreducing conditions. The monoclonal antibody YN1/1.7 that reacts with this antigen partially inhibits Con A stimulation of spleen cells, but its inhibition of LPA stimulation is negligible. Furthermore, the antibody profoundly inhibits MLR. The inhibition of MLR by YN1/1.7 antibody is comparable to that caused by anti-transferrin receptor. The time course study suggests that the antibody may directly inhibit proliferating cell populations in MLR. PMID- 2981918 TI - Antigen presentation by EBV-B cells to resting and activated T cells: role of interleukin 1. AB - We have previously demonstrated that Epstein Barr virus-transformed human B lymphocytes (EBV-B cells) present antigen to activated T cells (lines and clones) in a MHC-restricted manner. In the present study, using EBV-nonimmune donors, we demonstrate that EBV-B cells are unable to trigger tetanus toxoid (TT) antigen specific proliferation in autologous highly purified resting T cells. EBV-B cells from these same individuals were able to present TT to autologous TT-specific activated T cell blasts (Tbl). The inability of EBV-B cells to present TT to resting T cells was not caused by defective antigen processing by EBV-B cells. Thus, paraformaldehyde treatment of antigen-pulsed EBV-B cells did not impair their ability to trigger proliferation of antigen-specific Tbl, and EBV-B cells pulsed with antigen in the presence of autologous TT-specific T cell blasts did not present antigen to resting T cells. Furthermore, antigen-specific proliferation of resting T cells triggered by monocytes was enhanced rather than suppressed by EBV-B cells. The addition of partially purified human IL 1 allowed EBV-B cells to present TT antigen to resting T cells, suggesting that failure to secrete IL 1 contributed to the failure of EBV-B cells to present antigen. IL 1 could not be detected in supernatants of EBV-B cells stimulated with Staphylococcus epidermidis, concanavalin A, and TT antigen in the presence or absence of up to 5% autologous T cells. The differential capacity of EBV-B cells to present antigen to resting T cells vs activated T cells correlated with the T cell requirement for IL 1, because a rabbit antibody to human IL 1 inhibited the monocyte-supported proliferation of resting T cells but not that of activated T cells. These results suggest that the inability of EBV-B cells to present antigen to resting T cells is related to their inability to secrete detectable IL 1. PMID- 2981917 TI - Suppression of interleukin 2 receptor acquisition by monoclonal antibodies recognizing the 50 KD protein associated with the sheep erythrocyte receptor on human T lymphocytes. AB - Monoclonal antibodies OKT11A, 9.6, and 35.1 recognize epitopes on a 50000 dalton surface molecule (p50) identical to or closely associated with the sheep erythrocyte receptor (E receptor) on human T lymphocytes. These three antibodies were investigated for ability to inhibit T cell proliferation and interleukin 2 (IL 2) receptor acquisition (determined with anti-Tac antibody in an immunofluorescence assay) induced by the lectin mitogen phytohemagglutinin (PHA) or by the phorbol ester 12-O-tetradecanoyl-phorbol-13 acetate (TPA). OKT11A, 9.6, and 35.1 were found to suppress [3H]thymidine incorporation and IL 2 receptor acquisition stimulated by PHA but not by TPA. This inhibition was not attributable to a lag in kinetics, but was sustained throughout 4 to 5 days of culture. Because OKT11A and 9.6 have been reported to suppress lectin mitogen induced IL 2 production, we attempted to overcome inhibition of proliferation with exogenous IL 2 (MLA144 supernatants or immunoaffinity-purified human IL 2). Adding IL 2 at the initiation of culture abrogated the suppressive effect of all three anti-p50 antibodies on proliferation and on the acquisition of IL 2 receptors, raising the possibility that IL 2 may up-regulate expression of its cellular receptor on human T lymphocytes. These data, together with previous reports, indicate that OKT11A, 9.6, and 35.1 suppress lectin mitogen-induced T cell proliferation by impairing both IL 2 elaboration and IL 2 receptor acquisition, and suggest that IL 2 may be capable, at least under some conditions, of increasing expression of IL 2 receptors on human T lymphocytes. PMID- 2981919 TI - Functional properties of the 50 kd protein associated with the E-receptor on human T lymphocytes: suppression of IL 2 production by anti-p50 monoclonal antibodies. AB - Monoclonal antibody 9.6 is specific for a 50 kd T cell surface protein (p50) associated with the sheep erythrocyte (E)-receptor on human T lymphocytes. This antibody interferes with many T cell functions. We have examined the effect of antibody 9.6 on lymphocyte proliferation and interleukin 2 (IL 2) production triggered by mitogens, soluble antigens, and alloantigens to elucidate the mechanism(s) of its immunosuppressive action. At concentrations as low as 50 ng/ml, 9.6 suppressed lymphocyte proliferation and the elaboration of IL 2 by T cells stimulated by PHA, alloantigens, or low concentrations of the phorbol ester TPA (less than or equal to ng/ml). Furthermore, in cultures stimulated by a combination of PHA plus TPA, 9.6 did not inhibit the acquisition of IL 2 receptors but inhibited proliferation and IL 2 production. Immunoaffinity purified IL 2 completely restored lymphocyte proliferation in cultures inhibited by 9.6. Studies of kinetics of inhibition by 9.6 showed that this antibody inhibited lymphocyte proliferation induced by PHA, alloantigen, and PPD even when added at 24, 48, and 72 hr, respectively, after the initiation of these cultures, suggesting that 9.6 does not block lectin binding or antigen recognition by T cells and that it can inhibit lymphocyte proliferation even after cells have undergone one or more rounds of cell division. A dose-response analysis of lymphocyte proliferation induced by PHA or by TPA demonstrated that the degree of inhibition by 9.6 decreased with increasing concentrations of these mitogens. Antibody 9.6 did not inhibit lymphocyte response induced by optimal concentrations of PHA (50 to 100 micrograms/ml; PHA-M) but inhibited proliferation of maximally induced lymphocytes by using a synergistic combination of low concentrations of PHA (5 micrograms/ml, PHA-M) plus TPA (1 ng/ml). Taken together, these findings indicate that 1) 9.6 inhibits lymphocyte proliferation by affecting IL 2 production, 2) 9.6 does not inhibit the acquisition of 9.6 receptors induced by a synergistic combination of PHA plus TPA, and 3) p50 molecules may be involved in multiple pathways of T cell activation. PMID- 2981920 TI - Immortalization of BGDF (BCGF II)- and BCDF-producing T cells by human T cell leukemia virus (HTLV) and characterization of human BGDF (BCGF II). AB - Human peripheral T cells were transformed by human T cell leukemia virus (HTLV), and T cell lines producing BGDF (BCGF II) and BCDF were established. Among these cell lines, a cell line, TCL-Na1, secreted the highest level of both BGDF and BCDF, and the amount of BCDF secreted by TCL-Na1 cells was 900-fold more than that produced by PHA-stimulated T cells. Within the limits of our examination, none of the HTLV-transformed T cell lines produced IL 2 or BSF-p1 (BCGF I). BCDF produced by TCL-Na1 cells had a m.w. of 35,000 and a pI value of 5.5, being separated from BGDF, which was eluted in the fractions corresponding to m.w. of more than 60,000 and pI values of 5 to 6. BGDF induced both proliferation and IgM secretion in a mouse leukemic B cell line, BCL1, and these activities were not separated by either isoelectric focusing or gel filtration in the presence or absence of 0.1% Triton X-100, suggesting that the molecule designated BGDF exerted both growth and differentiation activities. BGDF acted on normal mouse B cells to induce proliferation as well as IgM secretion. The target cells of BGDF were in vivo activated B blast cells. BGDF acted on DXS-activated murine B cells to induce both proliferation and IgM secretion but not anti-Ig-activated B cells, indicating that BGDF and BSF-p1 were different molecules. PMID- 2981921 TI - Oligosaccharide structure of human C4. AB - The oligosaccharide structure of human C4 was studied by using C4 purified from plasma and C4 secreted by human hepatoma-derived cell line, HepG2. The alpha- and beta-chains of human C4 are glycosylated, whereas the gamma-chain is devoid of carbohydrate. The alpha-chain has three complex fucosylated oligosaccharides of the biantennary type, one each on the alpha 2, alpha 3, and alpha 4 fragments. The beta-chain has a single high mannose oligosaccharide primarily of the Man9GlcNAc2 type. The approximately 2000 Mr difference between the alpha-chains of the two C4 gene products (C4A and C4B) was localized to the alpha 2 fragment and is not due to carbohydrate. Sulfation of the C4 alpha-chain was localized to the alpha 4 fragment of the alpha-chain. Hence, the Mr difference between the two gene products is likely to reside in amino acid differences. The oligosaccharide structure of three incompletely processed C4 molecules was also analyzed. These molecules have the oligosaccharide composition of the appropriate individual subunits. Therefore, intracellular proteolytic processing to the multi-chain form of C4 is not required for proper oligosaccharide processing. PMID- 2981922 TI - Deactivation of guinea pig pulmonary alveolar macrophage responses to N-formyl methionyl-leucyl-phenylalanine: chemotaxis, superoxide generation, and binding. AB - Incubation of pulmonary alveolar macrophages (PAM) with the synthetic chemotactic tripeptide, N-formyl-methionyl-leucyl-phenylalanine (FMLP) results in deactivation of PAM chemotaxis. The chemotactic response to 10(-8) M FMLP was inhibited 85% after 30 min of preincubation with 10(-6) M FMLP and 48% by 10(-8) M FMLP. Only the higher dose of FMLP (10(-6) M) caused deactivation of the chemotactic response to C5a (20%). Preincubation with partially purified C5a at a concentration of 100 microliter/ml produced a 32% inhibition of the PAM response to 10(-8) M FMLP. In contrast, preincubation with FMLP had no significant effect on superoxide generation, either at baseline or after stimulation. Levels of intracellular cyclic adenosine-3',5'-monophosphate (cAMP) increased in response to PGE1 in the presence of 3-isobutyl-1-methylxanthine (IBMX), a phosphodiesterase inhibitor, but FMLP failed to induce a change in cAMP levels. Studies of 3H-FMLP binding were consistent with two populations of membrane receptors with different affinities. Preincubation of PAM with FMLP did not result in a reduction of maximal binding. We conclude that FMLP induces deactivation of PAM chemotaxis, but cross-deactivation occurs only after high dose treatment. Unlike the PMN, macrophage chemotactic activation is not accompanied by an elevation in cAMP levels. These observations suggest that PAM chemotaxis is influenced by prior exposure to chemotactic stimuli, but other aspects of the PAM response diverge from that of PMN. The mechanism of deactivation of PAM does not appear to result from a shift in the dose-response curve or decreased availability of membrane receptors, but may involve uncoupling of post-receptor cellular responses. PMID- 2981923 TI - Induction of cAMP-dependent protein kinase I during human monocyte differentiation. AB - Differentiation of human peripheral blood monocytes into macrophages was accompanied by induction of the regulatory subunit of cAMP-dependent protein kinase I as determined by photoaffinity labeling of cytosol proteins with 8-N3 [32P]cAMP and DEAE-Sephacel chromatography. The appearance of cAMP-dependent protein kinase I in macrophages was not due to translocation from the particulate fraction of monocytes. The regulatory subunit of cAMP-dependent protein kinase II was present in both monocytes and in vitro-differentiated macrophages. Protein kinase I in macrophages demonstrated higher affinity for 8-N3-cAMP (KD = 0.7 nM) than did protein kinase II from either monocytes (KD = 14.5 nM) or macrophages (KD = 4.9 nM). These studies demonstrate induction of the regulatory subunit of cAMP-dependent protein kinase I during the differentiation of a normal human cell and support the hypothesis that cAMP may regulate some stages of differentiation. PMID- 2981924 TI - Human endothelial cells modulate granulocyte adherence and chemotaxis. AB - Although human endothelial cells (EC) and granulocytes interact in several ways, the factors that regulate such interactions are not well defined. In this study we found that EC and their products directly altered granulocyte adherence (GA) and chemotactic activity. The spontaneous adherence of granulocytes to human umbilical vein EC monolayers was significantly reduced at 5, 15 and 30 min if the granulocytes were preincubated with EC that were stimulated by rocking. At 30 min the spontaneous adherence of EC-preincubated granulocytes was 36% of that of control granulocytes (P = 0.004). The augmented adherence stimulated by FMLP was also decreased (54% of control) by preincubation of the granulocytes with rocked EC. The ability of stimulated EC to inhibit GA was attenuated when the EC monolayers that were used for preincubation with the granulocytes were pretreated with the cyclooxygenase inhibitor indomethacin. GA to unstimulated EC was not significantly altered by indomethacin or aspirin, suggesting that cyclooxygenase products do not influence GA under resting conditions. Preincubation of granulocytes with rocked EC monolayers or supernatant media from rocked EC monolayers diminished their chemotactic response to FMLP by 45 to 65. This inhibition was also attenuated by pretreatment of the EC with indomethacin. EC supernatant medium caused a rapid increase in granulocyte intracellular cyclic AMP, with a maximum increase to 200% of control at 1 min. These data indicate that stimulated EC release one or more arachidonic acid products that alter spontaneous and inflammatory mediator-stimulated granulocyte activity. Prostacyclin, a major cyclooxygenase product of EC arachidonate, depressed inflammatory mediator-augmented GA to EC monolayers and chemotaxis when present in nanomolar concentrations. We conclude that EC-derived prostacyclin, alone or in combination with other EC products, alters GA and chemotaxis stimulated by inflammatory mediators. This provides a mechanism by which EC may modulate granulocyte distribution as well as granulocyte responses that are influenced by adherence, such as the release of toxic oxygen metabolites and granular enzymes. PMID- 2981925 TI - Oxidant membrane injury by the neutrophil myeloperoxidase system. I. Characterization of a liposome model and injury by myeloperoxidase, hydrogen peroxide, and halides. AB - Neutrophils and other phagocytes can injure cells by means of oxygen-dependent mechanisms, particularly the myeloperoxidase (MPO)-H2O2-halide system. The extent of such damage depends in part on the antioxidant defenses of the target cell. To facilitate the study of this phenomenon, we developed a model system in which we employed liposomes as targets for the myeloperoxidase system. The most useful species of liposomes employed 51Cr as the aqueous space marker and phosphatidyl choline with or without dicetyl phosphate and cholesterol as the structural lipid. Marker entrapment was established on the basis of 1) resolution of free from lipid-associated 51Cr by gel exclusion chromatography, 2) latency of 51Cr on rechromatography of detergent-treated liposomes, and 3) a correlation between entrapment and surface charge density. Exposure of liposomes to the complete MPO system resulted in release of 50 to 75% of the entrapped 51Cr. Release was abrogated by omission of myeloperoxidase or H2O2, heating of MPO, or addition of azide, cyanide, or catalase. Reagent H2O2 could be replaced by glucose plus glucose oxidase. Kinetic studies indicated a rapid process, lysis reaching half maximal levels in less than 2 min. The addition of cyanide at various times interrupted lysis at once, indicating a requirement for ongoing myeloperoxidase dependent reactions. Liposome disruption by the MPO system was pH dependent, increasing dramatically as pH was decreased from neutrality to 6.0. In the absence of halides, no lysis was observed. Maximum lysis was found with chloride at 10 to 100 mM, although at 1 mM concentrations, iodide, bromide, and thiocyanate were more active than chloride. Fluoride was inactive. Antagonism between halide species was demonstrated in that low concentrations of iodide or bromide inhibited the effect of optimal concentrations of chloride. Using 125I, we found that exposure of liposomes to the MPO system resulted in an association between iodide and liposomes; moreover, there was a close correspondence between this phenomenon and 51Cr release, suggesting that halogenation may be one mechanism of injury. These studies establish the usefulness of the liposome as a model of oxidant injury by a physiologically relevant system. They bear a striking parallel to work being done on MPO-mediated injury to eukaryotic and prokaryotic cells. By using this simplified model system, it should be possible to explore a number of determinants of target cell injury at a biochemical and molecular level. PMID- 2981926 TI - Oxidant membrane injury by the neutrophil myeloperoxidase system. II. Injury by stimulated neutrophils and protection by lipid-soluble antioxidants. AB - The stimulated human neutrophil can damage a variety of target cells, and in some models, a mechanism involving secretion of myeloperoxidase and H2O2 has been demonstrated. We explored the characteristics of this cell-cell interaction by using neutrophils and our recently described liposome model target cell system. Exposure of 51Cr-labeled liposomes to phorbol myristate acetate-stimulated human neutrophils resulted in release of 25 to 30% of the radioactivity. 51Cr release was abrogated by omission of the neutrophils, the phorbol ester or halide (iodide), replacement of the phorbol by an inactive congener, or addition of azide, cyanide, or catalase. Neutrophils from patients with hereditary absence of myeloperoxidase (MPO) or a failure of H2O2 formation (chronic granulomatous disease) did not cause liposome lysis unless purified MPO or a source of H2O2, respectively, was added. These data indicate that 51Cr release from liposomes is a consequence of the secretion of MPO and H2O2, which combine with extracellular halides to form a membrane lytic system. The influence of liposome composition on injury was then examined, with a focus on physiologically relevant lipid soluble antioxidants. Liposomes containing either alpha-tocopherol (0.33 to 1.67% of molar fraction of lipid) or beta-carotene (1.67% of molar fraction of lipid) were markedly resistant to lysis by the cellfree MPO-H2O2-chloride system. When the major structural lipid phosphatidyl choline was replaced by dipalmitoyl phosphatidyl choline, a synthetic phospholipid with no oxidizable double bonds, the resultant liposomes were totally resistant to lysis by the MPO-H2O2-chloride system. The addition of iodide to this system (i.e., both chloride and iodide present) changed the pattern of protection dramatically in that alpha-tocopherol and beta-carotene were no longer protective and the resistance of dipalmitoyl phosphatidyl choline liposomes was partial rather than complete. In contrast to iodide, the addition of bromide or thiocyanate did not have a major effect on the protection by antioxidants. Finally, we demonstrated protection by alpha tocopherol or dipalmitoyl phosphatidyl choline against liposome lysis by phorbol activated neutrophils. These studies illustrate the use of model phospholipid membranes in the characterization of oxygen-dependent cell-mediated cytotoxicity. Activated neutrophils lyse liposome targets through a MPO-dependent mechanism. Target properties, especially the content of lipid-soluble antioxidants, have a marked influence on susceptibility to lysis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2981927 TI - Removal or oxidation of surface membrane sialic acid inhibits formyl-peptide induced polymorphonuclear leukocyte chemotaxis. AB - Polymorphonuclear leukocyte (PMN) surface membrane glycoproteins are probably involved in the phenomenon of stimulus-response coupling. Consequently, we examined the effects of either removal or oxidation of surface membrane associated sialic acid residues on some responses of human PMN to chemotactic factors. Treatment of human PMN with either neuraminidase or sodium metaperiodate did not affect the ability of these cells to migrate randomly, but did inhibit their ability to respond chemotactically to the synthetic peptide N-formyl methionyl-leucyl-phenylalanine (FMLP). Treated PMN responded normally, however, to the complement-derived peptide C5a, and to the lipoxygenase product leukotriene B4. Enzymatic removal or oxidation of membrane sialic acid residues did not affect either FMLP-induced PMN degranulation or FMLP-induced generation by PMN of superoxide anion radicals. Removal of sialic acid did not significantly alter specific binding of [3H]FMLP to its receptor(s) on the PMN membrane. These findings indicate that sialic acid residues on the PMN surface membrane play an important role in modulating PMN responses to FMLP. PMID- 2981928 TI - Impaired oxidative burst does not affect human monocyte tumoricidal activity. AB - Human peripheral blood monocytes have been shown to lyse neoplastic cells selectively. It has been suggested that the oxidative burst may mediate monocyte tumoricidal activity. Two patients with chronic granulomatous disease were studied. The first patient had no detectable secretion of hydrogen peroxide and superoxide anion after stimulation with 100 ng/ml PMA. Nevertheless, his tumoricidal activity measured against K562 targets in 4-hr 51Cr-release assays by using highly purified monocytes was reproducibly above the 95th percentile of the normal population. His monocytes lysed WEHI-164 targets pretreated with 1 microgram/ml Actinomycin D with similar efficiency. Another patient whose oxidative burst was 15% of normal nevertheless killed Daudi targets efficiently. In addition, oxygen-deprived monocytes of normal donors manifested normal tumoricidal activity, despite failure to produce any detectable oxidative burst after PMA stimulation. We conclude that reactive oxygen metabolites are not the primary mediators of tumor cytolysis by human peripheral blood monocytes. PMID- 2981929 TI - Human mononuclear phagocyte antiprotozoal mechanisms: oxygen-dependent vs oxygen independent activity against intracellular Toxoplasma gondii. AB - To determine if the oxygen-dependent and -independent antiprotozoal mechanisms with which the human mononuclear phagocyte is equipped to act against Leishmania donovani operate against other intracellular parasites, oxidatively intact and deficient cells were challenged with Toxoplasma gondii. Fresh monocytes and lymphokine- or gamma-interferon (IFN-gamma)-activated macrophages from normal individuals killed 35% and 50% of T. gondii within 6 hr, respectively, and each of these cell populations inhibited the replication of surviving parasites 20 hr after infection. This activity was associated with the capacity to release large amounts of H2O2 (572 to 971 nmol/mg) and to respond to toxoplasma ingestion with respiratory burst activity. Impairing the ability to generate oxygen intermediates by glucose deprivation or treatment with superoxide dismutase, catalase, or mannitol inhibited toxoplasmacidal activity by greater than 80% and permitted a 2.6- to 4.3-fold increase in the number of intracellular toxoplasmas. In contrast to normal cells, fresh monocytes from patients with chronic granulomatous disease (CGD) killed less than 8% of toxoplasmas and exerted 50% less toxoplasmastatic activity. However, although associated with the induction of only modest toxoplasmacidal effects (18 to 20% killing), lymphokine stimulation did induce CGD monocytes and macrophages as well as oxidatively inactive human endothelial cells to display near normal levels of toxoplasmastatic activity. Similar to oxygen-dependent mechanisms, the enhancement of oxygen-independent activity by crude lymphokines could be abolished by a monoclonal anti-IFN-gamma antibody and could be achieved by treatment with recombinant IFN-gamma alone. Unstimulated CGD monocytes, however, were found to lose all antitoxoplasma activity after two days in culture, whereas normal cells continued to effectively inhibit T. gondii replication, suggesting that oxygen-independent responses may not actually be required for the normal monocyte to act against T. gondii. Taken together with previous findings with L. donovani, these results indicate that the human mononuclear phagocyte possesses an oxygen-independent antiprotozoal mechanism and that its effects can be enhanced by lymphokines (IFN-gamma), but that nevertheless this cell's primary response to intracellular protozoa is largely oxygen dependent. PMID- 2981930 TI - Sensitive ELISA for IgG and IgM anti-albumin autoantibodies not influenced by HBsAg-associated polyalbumin receptors. AB - An enzyme immunoassay for the detection of IgG and IgM anti-polymerized albumin autoantibodies (AAA) is described. It was found that polyalbumin receptors on HBsAg particles interfere in the detection of IgG AAA when polymerized human albumin (pHSA), but not polymerized bovine albumin (pBSA), is used as coating antigen. Polyalbumin receptors do not appear to interfere in the detection of IgM AAA, with either pHSA or pBSA as coating antigen. All normal sera showed evidence of AAA, of both IgG and IgM classes. Levels of IgG and IgM AAA in sera from most type A and type B acute hepatitis patients were above the range of normal controls. ELISA detection of AAA distinct from HBsAg reactivity can help in understanding the role of these autoantibodies in HBV infection. PMID- 2981931 TI - Immunoassay for the detection and quantitation of infectious human retrovirus, lymphadenopathy-associated virus (LAV). AB - Detection of replicating human retroviruses has relied upon rather cumbersome reverse transcriptase, immunofluorescence, or electron microscopic assays. We describe a new sandwich enzyme-linked immunoassay (ELISA) for detecting the human retrovirus, lymphadenopathy-associated virus (LAV), in supernates of LAV-infected human lymphocyte cultures. This LAV capture immunoassay compares favorably with the reverse transcriptase assay, despite the fact that it is performed on 20-fold less supernate material. Because the assay can be performed on 0.1 ml of culture supernate and is done by an ELISA method, LAV inoculation of lymphocyte cultures can be monitored quite conveniently, and endpoint titrations of infectious virus (ID-50 assays) can be performed. We demonstrate the application of the capture assay and ID-50 assay to disinfectant and serum neutralization experiments. PMID- 2981932 TI - Reduction in non-specific binding in enzyme immunoassays using casein hydrolysate in serum diluents. PMID- 2981933 TI - Effects of synthetic leukotrienes on local blood flow and vascular permeability in porcine skin. AB - The local effects of synthetic leukotrienes (LT) were examined in the skin of the anesthetized pig. Blood flow was measured noninvasively with the use of a laser Doppler flow meter and changes in vascular permeability were measured using technetium-labeled human serum albumin as a marker for extravasation. LTB4 and the peptidolipid leukotrienes, LTC4, LTD4, LTE4, LTF4, induced vasodilator responses when injected intradermally at a dose of 1 ng. The vasodilator effects of LTB4 and LTF4 were comparable in magnitude to those of prostaglandin E2 (PGE2) and histamine and persisted over a wide dose range. Vascular permeability was induced by histamine, PGE2, and LTB4 but not by the other leukotrienes. The effects of LTB4 were significantly increased in the presence of PGE2. Leukotrienes appear not to produce their effects through the generation of prostaglandins as neither the vasodilator nor the permeability-enhancing effects were affected by treatment with indomethacin. The present investigation demonstrates that the pig is the first animal model to be described which reflects the potent vasodilator actions of leukotrienes in human skin. The porcine skin may thus be a useful model in the study of human skin diseases. PMID- 2981934 TI - Molecular epidemiology of transmissible gentamicin resistance among coagulase negative staphylococci in a cardiac surgery unit. AB - The prevalence of colonization of patients in a cardiac surgery unit with gentamicin-resistant, coagulase-negative staphylococci increased from 20% to 68% over a period of four years. Gentamicin resistance was found to be plasmid associated and transmissible from wild-type coagulase-negative staphylococcal donors to a Staphylococcus aureus recipient by filter mating (conjugative). These plasmids were present in isolates from 50 (74%) of 69 individuals examined. This figure included isolates from colonized patients, colonized staff, and patients with prosthetic valve endocarditis. A common restriction-endonuclease digestion pattern (pG02; 50 kilobases) was seen in only 19 (38%) of the 50 conjugative plasmids. However, filter hybridization, restriction-endonuclease mapping, and transposon insertional mutagenesis showed that representatives of the other 10 restriction-endonuclease digestion patterns were physically related to pG02 over greater than 80% of their genome, with differences largely due to deletions or insertions of DNA in three areas, and that their gentamicin-resistance genes were identical. Molecular analysis may be required to ascertain the physical similarity among phenotypically and epidemiologically related plasmids. PMID- 2981935 TI - Natural killer cell activity in infants and children excreting cytomegalovirus. AB - Natural killer cell (NKC) activity against uninfected and cytomegalovirus (CMV) infected fibroblasts and K562 cells was assessed in 39 infants and children excreting CMV and 89 controls. NKC activity in cord blood specimens, healthy donors up to six months old, and donors older than six months showed significant increases (P less than .02) with age. NKC activities of CMV excreters correlated with acute disease characterized by interstitial pneumonitis (IP) and showed differential responses to the three target cells. CMV excreters younger than six months with IP showed greater mean percentage lysis (MPL; 36.6% +/- 10.5%) of CMV infected cells than did those without pneumonitis (25.9% +/- 10%) or controls (22.1% +/- 8.0%). Excreters older than six months with IP showed activities comparable to controls; NKC activities of excreters without IP were depressed. Significantly elevated activity against uninfected fibroblasts characterized patients with IP regardless of age (MPL, 30.2% +/- 7.3% vs. 4.2% +/- 2.0% for healthy controls and 6.3% +/- 5.4% for excreters without IP). Increased cytolysis of uninfected fibroblasts was therefore associated with more active disease. PMID- 2981936 TI - Rapid diagnosis of cytomegaloviral pneumonia by tissue immunofluorescence with a murine monoclonal antibody. AB - A murine monoclonal antibody to cytomegalovirus (CMV) was used to identify virus infected cells in coded frozen tissue sections from 52 consecutive open-lung biopsies obtained from marrow transplant recipients with pneumonia. The diagnostic sensitivity of immunofluorescence (IF) using this antibody exceeded that of standard histology performed on touch imprints and frozen and permanent lung sections and was equal to viral culture and in situ CMV nucleic acid hybridization. In comparison with patients with CMV pneumonia demonstrated histologically and by IF, those with negative histology and positive IF were more likely to have seroconverted before biopsy. Despite this evidence of an immune response to CMV pneumonia, the two groups did not differ in CMV positivity by culture or by hybridization, and their long-term survival was equally poor. The findings demonstrate that this antibody can play an important role in both the rapid diagnosis of CMV infection and the elucidation of CMV pathophysiology. PMID- 2981937 TI - Cytomegalovirus infection in sex partners: evidence for sexual transmission. AB - To examine the hypothesis that cytomegalovirus (CMV) is sexually transmitted, we determined the prevalences of antibody to CMV and viral shedding in 63 male sex partners of women with or without CMV infection, and CMV isolates from infected couples were compared by DNA restriction enzyme analysis. The prevalence of seropositivity by enzyme-linked immunosorbent assay was 31 (74%) of 42 men whose female partners were seropositive compared with five (31%) of 16 men whose partners were seronegative (P = .008). CMV was isolated from the semen or urine of four (22%) of 18 men whose female partners shed CMV from the cervix or urine compared with none of 42 whose partners were culture negative (P = .013). DNA restriction enzyme typing of CMV isolates from three pairs of sex partners showed that two of the couples were infected with common strains; epidemiologically unrelated isolates gave distinct patterns. Heterosexual contact is a major mode of transmission of CMV among some young adults. PMID- 2981938 TI - Experimental model for activation of genital herpes simplex virus. AB - We have been actively studying herpesvirus latency and the effect of antiviral agents on latent infections in vivo and in vitro. To establish a model of recurrent herpetic genital disease, we treated latently infected C57Bl/6 mice with immunosuppressive agents. Mice were injected intraperitoneally with cyclophosphamide and antilymphocyte serum and then were examined for reactivated virus or recurrent lesions. When latently infected mice were subjected to immunosuppression, neither recurrent lesions nor virus was detected at the site of primary infection. However, when mice were selected for immunosuppressive treatment by the presence of postinfection scars, recurrent disease was induced. The results point to the significance of local factors in the pathogenicity of recurrent herpetic genital disease in mice. PMID- 2981940 TI - High-dose oral acyclovir for children at risk of disseminated herpesvirus infections. PMID- 2981939 TI - The origin of the HM175 strain of hepatitis A virus. PMID- 2981941 TI - Iron deficiency anemia: mitochondrial alpha-glycerophosphate dehydrogenase in guinea pig skeletal muscle. AB - Severe iron deficiency anemia in rats causes a decrease in the activities of iron containing enzymes in skeletal muscle mitochondria, and subsequent diminished respiratory activity has been linked to lowered work capacity. It was suggested that loss of mitochondrial alpha-glycerophosphate dehydrogenase activity plays a particularly important role in this process and, by inference, in the clinical manifestations of iron deficiency anemia. This view may be ill founded, inasmuch as other pathways with potentially greater activity are capable of transporting reducing equivalents from the cytosol into the mitochondria in mammalian skeletal muscle. In our experiments, iron deficiency anemia of a severity on the order of that in humans was produced in guinea pigs. Mitochondria from skeletal muscles of test animals exhibited respiration rates diminished by 24% to 36% compared with control mitochondria in the presence of several substrates. However, differences in respiration were not observed with alpha-glycerophosphate as substrate, nor were there differences in alpha-glycerophosphate dehydrogenase enzyme activity between mitochondria from iron-deficient and control animals. Although cytochrome oxidase activity and muscle mitochondrial protein content were the same in both groups of guinea pigs, cytochrome and flavoprotein concentrations were lower in mitochondria from iron-deficient animals and there was a preferential loss of cytochrome c + c1. Iron deficiency anemia in guinea pigs thus results in impaired oxygen metabolism in skeletal muscle mitochondria that is associated with a general decrease in the concentrations of iron-containing electron transport chain components as well as with an alteration in chain stoichiometry.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981942 TI - Nitrofurantoin-stimulated oxidant production in pulmonary endothelial cells. AB - Nitrofurantoin, a urinary antiseptic, is associated with significant pulmonary toxicity. Our study indicates that nitrofurantoin may produce lung injury by directly stimulating lung parenchymal cells to generate toxic oxygen species such as superoxide and hydrogen peroxide, which can overwhelm cellular antioxidant defenses and result in permanent injury to the cells. Nitrofurantoin (10(-3) mol/L) stimulates bovine pulmonary artery endothelial cells to release 3.7 +/- 0.4 mumol/L superoxide per 10(5) cells and 4.4 +/- 0.5 mumol/L hydrogen peroxide per 10(5) cells (p less than 0.001 compared with endothelial cells without nitrofurantoin). Endothelial cells treated with nitroblue tetrazolium, a yellow dye reduced by superoxide to insoluble blue formazan, can be monitored both spectrophotometrically and morphologically. Nitrofurantoin (10(-5), 10(-4), and 10(-3) mol/L) stimulated pulmonary endothelial cells to reduce nitroblue tetrazolium monitored spectrophotometrically as 0.022 +/- 0.001, 0.032 +/- 0.002, and 0.071 +/- 0.004 delta A515 per 10(5) cells per hour, respectively (p less than 0.001, comparison of cells with 10(-4) and 10(-3) mol/L nitrofurantoin vs. control cells. From the same dose-response curve, endothelial cells incubated with nitrofurantoin morphologically demonstrated formazan granules in the cytoplasm in 17% +/- 6%, 71% +/- 9%, and 92% +/- 5% of cells. Nitrofurantoin (10( 5), 10(-4), and 10(-3) mol/L) directly injures 51Cr-labeled pulmonary endothelial cells, with injury expressed as a cytotoxic index of 1 +/- 1, 20 +/- 4, and 51 +/ 3, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981943 TI - Synovial sarcomas of the neck. A report of two cases. AB - Two cases of synovial sarcomas in the hypopharynx are reported, together with a review of similar neoplasms previously reported in this unusual region. The treatment of choice seems to be aggressive, with radical surgery followed by radiation therapy. PMID- 2981944 TI - Influence of prostaglandin I2 on fibronectin-mediated phagocytosis in vivo and in vitro. AB - This study evaluated the effect of prostaglandin I2 (PGI2) on fibronectin mediated macrophage phagocytosis in vivo and in vitro. Phagocytosis measured in vivo in rats by the vascular clearance rate and hepatic localization gelatinized sheep erythrocytes was inhibited in a dose-dependent manner after intravenous administration of PGI2. Phagocytosis was assessed in vitro in terms of uptake of fibronectin-dependent gelatinized sheep erythrocytes by monolayers of casein elicited rat peritoneal macrophages. Concentrations of 1 ng/ml PGI2 or greater resulted in inhibition of particle internalization but not attachment to macrophages. This inhibitory effect was enhanced by aminophylline, a phosphodiesterase inhibitor. PGI2 increased cAMP levels and these were further increased in the presence of aminophylline. These data indicate that PGI2 inhibits macrophage uptake of gelatinized particles and support the idea that this is mediated by increased intracellular levels of cyclic AMP. PGI2 should thus be considered a potential etiologic factor in the phagocytic depression observed in association with thrombosis. PMID- 2981945 TI - The response of human peripheral blood mononuclear phagocytes to rheumatoid arthritis. AB - The maturity of peripheral blood mononuclear phagocytes (B-MPs) from patients with rheumatoid arthritis (RA), osteoarthritis (OA), and reference ("normal") subjects was compared. Mononuclear cell isolates from peripheral blood were separated on discontinuous gradients of Percoll into low density (more mature) and high density (less mature) subpopulations. Contrary to expectations, the proportion of immature B-MPs in RA patients was found to be significantly lower than that in reference subjects. In RA patients with synovial effusions the proportion of immature B-MPs approached but did not exceed those found in reference subjects, despite the fact that 31% of these patients displayed a peripheral blood monocytosis. It was concluded that the bone marrow precursor population had adapted to the long-term demand for B-MPs in the course of this chronic inflammatory disease. PMID- 2981946 TI - Expression of 5'nucleotidase activity and wheat-germ agglutinin binding sites in mononuclear phagocytes from bone marrow cultures. AB - The question as to whether the various types of mononuclear phagocyte found in bone marrow cultures and recognized by specific peroxidatic (PO) activity patterns differ in the expression of binding sites for the lectin wheat-germ agglutinin (WGA) and the activity of the ectoenzyme 5'nucleotidase (5'N) was investigated. Monoblasts, promonocytes, monocytes, and/or exudate macrophages, and exudate-resident macrophages generally showed a high level of WGA binding, and a considerably lower level was found in the PO-negative cells and in resident macrophages. 5'N activity was absent in monoblasts, promonocytes, and in the great majority of the monocytes and/or exudate macrophages, but was demonstrable in exudate-resident macrophages and resident macrophages, as well as in PO negative macrophages after 4 days of culture. On the basis of the successive occurrence of the above-mentioned phenotypes in cultures, the possibility that this diversity in WGA binding and 5'N activity is related to modulation during cell differentiation is discussed. The present findings led to the conclusion that the PO-negative macrophages, whose origin was previously not entirely certain, are precursors of resident macrophages. PMID- 2981948 TI - Changing concepts: treating preinvasive cervical neoplasia. PMID- 2981947 TI - Enzyme histochemical comparison of biomphalaria glabrata amebocytes with human granuloma macrophages. AB - In fresh water snails, amebocytes are the principal cells that react to parasitic infection. Ultrastructurally, amebocytes resemble mammalian macrophages. To clarify the relationship between amebocytes and macrophages, we compared the histochemical staining for seven enzymes in Biomphalaria glabrata snail amebocytes, both in the amebocyte-producing organ (APO) and in the encapsulation reaction formed around parasite sporocysts with the staining in macrophages from the lymph nodes of patients with sarcoid or tuberculosis. Snails were infected with Echinostoma paraensei and Schistosoma mansoni miracidia. APOs and ventricular tissue with encapsulated parasites were fixed and embedded in glycol methacrylate monomer. Hardened blocks were sectioned at 2 micron and stained for alkaline phosphatase, acid phosphatase, alpha-naphthyl acetate esterase (ANAE), ATPase, peroxidase, 5'nucleotidase, and chloroacetate esterase. The amebocyte producing organ contained cells that were positive for acid phosphatase, ANAE, and ATPase. Amebocytes in the capsules formed around echinostome sporocysts showed stronger staining for the same three enzymes. Capsules did not form around schistosome sporocysts, but the connective tissue around them contained numerous amebocytes that were also positive for these three enzymes. The amebocyte enzyme histochemistry resembled that in human granuloma macrophages, but differed from that in neutrophils. The increased expression of enzymes in amebocytes involved in the encapsulation reaction as compared to those in the APO was reminiscent of the alterations observed when human monocytes convert to tissue macrophages. These studies support the hypothesis that the amebocyte is an "invertebrate macrophage." PMID- 2981949 TI - Lactic acid and steroid production by intact mouse adrenal glands and cell suspensions: effects of nucleotide derivatives and substrates. AB - The effects of the dibutyryl derivatives of cyclic GMP and cyclic AMP on lactic acid and steroid production were compared in intact mouse adrenal glands at concentrations of 0.5-1 mmol/l and in mouse adrenal cell suspensions at concentrations of 0.01-1 mmol/l. The dibutyryl derivative of cyclic GMP had little or no effect on lactic acid production in either tissue preparation. It caused a slight stimulation of corticosteroid output in intact glands at a concentration of 1 mmol/l, amounting to one-tenth of the response observed with 1 mM-dibutyryl cyclic AMP. Dose-dependent increases in lactic acid and steroid production were obtained with dibutyryl cyclic AMP in cell suspensions. AMP and GMP increased lactic acid but not steroid production. All the substrates tested (glucose, glucose-6-phosphate, fructose, fructose-6-phosphate, fructose-1,6 diphosphate, 10 mmol/l; pyruvate and glycerol, 20 mmol/l) stimulated basal glycolysis in intact glands and cell suspensions and none affected basal steroid production significantly. By far the greatest increase in lactic acid production was noted with fructose-1,6-diphosphate. However, only glucose and, in unsectioned glands, pyruvate exerted a potentiating effect on the glycolytic response to ACTH. Glucose potentiated the steroidogenic response to ACTH also, but only in intact glands. The relative ineffectiveness of dibutyryl cyclic GMP is in accord with the species-dependent differing responses to the free form of the cyclic nucleotides noted in mouse and rat adrenal glands. The substrate requirements are in keeping with a rate-limiting role of phosphofructokinase and an action of ACTH at some site between the entry of glucose into the cell and the formation of fructose-1,6-diphosphate. PMID- 2981950 TI - Influence of growth hormone, corticosterone, corticotrophin and changes in the environmental temperature on pituitary-ovarian function in the immature rat. AB - The influence of various endocrine and environmental factors on pituitary-ovarian function was studied in peripubertal rats treated with pregnant mare serum gonadotrophin (PMSG). Pregnant mare serum gonadotrophin induced ovulation in rats aged 27 days provided they weighed over 60 g. The response was preceded by a marked hypersecretion of LH which was detectable by radioimmunological and biological assay methods. In contrast, smaller rats of the same age did not ovulate in response to PMSG apparently because of the secretion of a pleiomorphic form of LH which, although immunoreactive, appeared to be biologically inactive. Ovarian function, assessed by response to exogenous gonadotrophins and by measurement of 125I-labelled human chorionic gonadotrophin binding, was normal despite the presence of the biologically inactive pleiomorph. Exposure of the small PMSG-treated rats to a high environmental temperature (39 degrees C) or treatment with corticosterone or GH altered the nature of the LH in the blood so that it was active in both assay systems and facilitated ovulation as also did ACTH. The results suggest that the abrupt change in the nature of the LH released by the pituitary gland essential for the initiation of ovulation may be affected by GH, corticosterone or a raised environmental temperature. PMID- 2981951 TI - Human malignant T cells capable of inducing an immunoglobulin class switch. AB - Evidence is presented for the existence of a "switch" T cell derived from a patient with mycosis fungoides/Sezary's syndrome. The serum immunoglobulin profile in this patient revealed high IgG and IgA but no detectable IgM. Peripheral blood mononuclear cells from this patient secreted only IgG and IgA in the presence of pokeweed mitogen. T cells (Trac) co-cultured with normal allogeneic non-T cells and pokeweed mitogen resulted in only IgG and IgA PFC, with little or no IgM secretion. There was no evidence of active suppression of IgM. Rather, these T cells appeared to induce an Ig class switch from IgM to IgG and IgA, when co-cultured with mu+ tonsillar B cells. Further evidence was obtained using mononuclear cells derived from a patient with immunodeficiency and hyper-IgM, a syndrome characterized by a lack of IgG and IgA secretion. The addition of Trac cells to either peripheral blood mononuclear cells or non-T cells from a patient with hyper-IgM syndrome resulted in new secretion of IgG, with a concomitant decrease in IgM secretion, whereas control T cells were not effective in inducing secretion of any isotype other than IgM. Isolated Tac+ T cells from Trac appear to be responsible for this effect. PMID- 2981952 TI - Interleukin 2 receptors on human B cells. Implications for the role of interleukin 2 in human B cell function. AB - In the present study, we examined the expression of interleukin 2 (IL-2) receptors on normal human B cells as well as established B cell lines. Anti-Tac monoclonal antibody did not bind to freshly separated normal human B cells. Unexpectedly, with the appropriate activation of the normal B cells by anti-mu antibody, phorbol myristate acetate, or Staphylococcus aureus Cowan I (SAC), Tac antigen was induced on the activated B cells. Anti-Tac antibody showed consistent reactivity with two B cell lines that were infected by human T cell leukemia virus (HTLV) and some reactivity with two out of eight Epstein-Barr virus transformed B cell lines established from normal adult donors. Immunoprecipitation analysis revealed that antigens of similar size with a molecular weight of 50,000-60,000 can be precipitated with anti-Tac antibody from phytohemagglutinin-stimulated normal T cell blasts and normal activated B cells, as well as a cloned B cell line. Binding assays of IL-2 on normal activated B cells and on the cloned B cell (HS1) revealed that B cells have significantly fewer sites and lower-affinity IL-2 receptors compared with phytohemagglutinin stimulated normal T cell blasts. Finally, biological properties of the IL-2 receptor on B cells were examined by incubating B cells with recombinant IL-2. It was found that moderate concentrations of IL-2 induce significant enhancement of proliferation and differentiation in SAC-activated normal B cells. These results suggest that normal B cells may express functional IL-2 receptors or closely related proteins and thus IL-2 may play a significant role in the modulation of B cell function. PMID- 2981954 TI - Adoptive transfer studies demonstrating the antiviral effect of natural killer cells in vivo. AB - We carried out adoptive transfer studies to determine the role of natural killer (NK) cells in resistance to murine cytomegalovirus (MCMV) and lymphocytic choriomeningitis virus (LCMV). We transferred leukocytes from adult mice into suckling mice 1 d before injecting them with virus. Resistance was measured by enhancement of survival and reduction of virus multiplication in the spleens of recipient mice. The phenotype of the cell population capable of mediating resistance to MCMV was that of a nylon wool-nonadherent, asialo GM1+, NK 1.2+, Ly 5+, Thy-1-, Ia-, low density lymphocyte; this is the phenotype of an NK cell. Cloned NK cells, but not cloned T cells, provided resistance to MCMV in suckling mice. Cloned NK cells also provided resistance to MCMV in irradiated adult mice, and antibody to asialo GM1, which depletes NK cell activity in vivo, enhanced the synthesis of MCMV in athymic nude mice. Neither adult leukocytes nor cloned NK cells influenced LCMV synthesis in suckling mice. We conclude that a general property of NK cells may be to provide natural resistance to virus infections, and that NK cells can protect mice from MCMV but not from LCMV. PMID- 2981953 TI - Mechanism of neutralization of influenza virus by secretory IgA is different from that of monomeric IgA or IgG. AB - We have found that bile is a useful source of secretory IgA (scIgA) which can specifically neutralize influenza virus infectivity. Using purified scIgA, we compared the mechanism of neutralization with that mediated by IgA monomers (prepared from scIgA by differential reduction) and IgG. At 4 degrees C, scIgA prevented the attachment of neutralized virus, while neither monomeric IgA nor IgG had any affect on this process or on the subsequent stages of infection by which virion RNA accumulates in nuclei. At 25 and 37 degrees C, scIgA permitted the attachment of approximately half the neutralized virus, but the virus was not internalized. Clearly, the neutralization depends on the character of the antibody used. scIgA may act by steric hindrance (with attachment or penetration, depending on temperature), whereas IgA and IgG neutralize infectivity at a stage subsequent to accumulation of the virus genome in the nucleus. PMID- 2981955 TI - Irritable bowel syndrome. PMID- 2981956 TI - Interaction of Ross River virus with the complement system. AB - In the absence of virus-specific antibody, Ross River virus failed to activate either the classical or alternative complement pathways. Instead, it inhibited the cleavage of C3 via both pathways. The virus did not appear to act by disrupting C3bBb complexes or by preventing cleavage of factor B by factor D. Instead Ross River virus was found to interfere with the actual cleavage of C3 by activated factor B (C3bBb) of the alternative pathway and C4b2a of the classical pathway. PMID- 2981957 TI - Ultrastructural localization of the herpes simplex virus major DNA-binding protein in the nucleus of infected cells. AB - Using immunocytochemical procedures in conjunction with electron microscopy we have examined the distribution of the major DNA-binding protein (DBP) of herpes simplex virus (HSV) in infected nuclei. In embedded specimens, DBP was preferentially associated with fibrillar material of electron-translucent viral inclusions, and to a lesser extent with peripheral (marginated) host chromatin. The latter association was sensitive to a non-ionic detergent ('Photo flo'). In chromatin spread by the Miller technique, DBP was found to be a component of the 10 nm 'thick filaments' previously described in HSV infection. PMID- 2981958 TI - Nucleotide sequence of bovine papillomavirus type 2 late region. AB - The late region of bovine papillomavirus type 2 (BPV-2) DNA has been identified. The complete nucleotide sequence of the region was determined and revealed two large open reading frames. The DNA sequence results and the predicted amino acid sequence of putative polypeptides encoded by this region are presented. Comparative analysis of the BPV-2 late region and the corresponding area of BPV-1 was performed. This study demonstrates that identical genetic organization and considerable nucleotide sequence conservation exists between these two serotypes. PMID- 2981959 TI - Mutagenesis of a herpes simplex virus origin of DNA replication and its effect on viral interference. AB - Supercoiled plasmid molecules containing cloned copies of a DNA fragment which includes a functional herpes simplex virus type 1 origin of DNA replication were cleaved preferentially at two positions within the viral insert by nuclease S1. Plasmids with molecular linker insertions at these sites were constructed, and analysis of two representative plasmids demonstrated the presence of palindromic DNA sequences at the preferred cleavage positions. One of these palindromic sequences occurred within a 90 bp region in which the cis-acting sequences essential for viral origin function had previously been located. Insertion of a linker at this position abolished origin activity, demonstrating an essential role for sequences within the palindrome in the initiation of DNA synthesis. In transfection assays, plasmids containing a functional viral origin of DNA replication markedly interfered with the infectivity of non-defective viral DNA even in the absence of viral encapsidation signals. Inactivation of the origin greatly reduced this effect on DNA infectivity, suggesting that viral interference may be mediated by a mechanism involving the replicative machinery. PMID- 2981960 TI - Distribution of G + C-rich regions in varicella-zoster virus DNA. AB - The distribution of G + C-rich sequences in the genome of varicella-zoster virus (VZV) was investigated by partial denaturation, equilibrium sedimentation and Southern blot analyses. Portions of the IRS and TRS repeat sequences bounding the US region of the DNA were found to have a G + C content 10 to 20% greater than the overall 47% G + C content of the VZV genome. A stretch of DNA (approx. 1500 base pairs) at the UL-IRS junction and repeated at the terminus of the TRS sequences was found to be about 64% G + C, based on sedimentation equilibrium measurements. We also report the cloning of a novel fragment containing sequences from both the UL and TRS termini of the VZV genome. Our ability to clone this fragment suggests that unusual forms of VZV DNA including closed circular molecules and molecules with an inverted UL region can be packaged into nucleocapsids. PMID- 2981961 TI - Analysis of bovine cytomegalovirus genome structure: cloning and mapping of the monomeric polyrepetitive DNA unit, and comparison of European and American strains. AB - The genome of bovine cytomegalovirus (BCMV) strain 66-P-347 consists of double stranded, linear DNA with a size of 144 +/- 6 kb. It contains polyrepetitive DNA (prDNA) segments like five other BCMV strains. The restriction patterns of the prDNA of all six strains are very similar and indicate that monomeric prDNA units are either 1950 bp (class I and Ia), 2350 bp (class II) or 2750 bp (class III) in size. The complete unit sequence of strain 66-P-347 (class II) was cloned in bacteriophage vector M13mp7 and mapped by the restriction enzymes EcoRI, BamHI, Bg/I, NaeI, SstII and PstI. From these results the restriction maps of the prDNA of the other five strains were deduced. The 400 bp differences in size between the three prDNA classes are a consequence of the appearance of an internal 200 bp sequence being present one-, three- or fivefold in head-to-tail formation. Hybridization of 35S-labelled recombinant phage DNA to Southern blots with DNA of the different strains leads to the conclusion that prDNA units are present as multimers in tandem formation at both genomic termini in the same orientation. The number of terminal repeat units varies between individual molecules of a strain, but the actual terminal sequences are identical. PMID- 2981962 TI - Genome differences among field isolates and vaccine strains of pseudorabies virus. AB - The DNA of field isolates and vaccine strains of pseudorabies virus (PRV) was analysed by digestion with the restriction endonuclease BamHI. A number of distinct restriction profiles of the field isolates obtained from different locations within Europe were observed. As for herpes simplex virus, the variations could be classified into two types: first, alterations in the mobility of fragments due to the presence of additional sequences and/or the occurrence of deletions, a phenomenon most apparent in fragments containing part or whole of the repeat sequence of PRV DNA; second, generation of differently sized fragments due to loss and/or gain of restriction endonuclease cleavage sites. By analysis of several strains with BamHI a small number of variable cleavage sites were identified within particular regions of the unique long (UL) segment of the PRV genome. Compared to wild-type PRV, the restriction fragment patterns of vaccine strains showed characteristic alterations, including the absence of bands, which were non-variable in wild-type strains, and/or the presence of new bands some of which were submolar. Some of these characteristics could be explained by a deletion in the unique short (US) region of the genome of most vaccine strains and the occurrence of closely related variants in the uncloned vaccine virus stocks. PMID- 2981963 TI - Further characterization of the in vitro products generated by proteolytic cleavage of Gazdar murine sarcoma virus p65gag. AB - In vitro proteolytic cleavage of the Gazdar murine sarcoma virus (Gz-MuSV) p65gag polypeptide (Gz-p65gag) was facilitated by detergent-disrupted Moloney murine leukaemia virus (Mo-MuLV). Incubation of radioactively labelled Gz-p65gag in the presence of unlabelled Mo-MuLV under optimal conditions resulted in the cleavage of Gz-p65gag to proteins of 40000 (P40) and 25000 (P25) Mr. P40 and P25 appeared to be similar in both mobility and antigenicity to Mo-MuLV intermediates, Pr40gag and Pr25gag, previously found in infected cells. Additional proteins of 30000 (Gz p30), 15000 (Gz-p12), 12000 (Gz-p15) and 10000 (Gzp10) Mr were also generated upon cleavage of Gz-p65gag and contained antigenic determinants of Mo-MuLV structural proteins p30, pp12, p15 and p10, respectively. Both detergent disrupted Mo-MuLV and Rauscher murine leukaemia virus produced similar cleavage profiles. Trypsin and detergent-disrupted mouse mammary tumour virus generated cleavage patterns very different from that produced by Mo-MuLV. Both visual and quantitative time studies of the reaction indicated that P40 gave rise to Gz-p30 and Gz-p10. Tryptic peptide mapping of Gz-p65gag and its cleavage products supported the results obtained from both immunoprecipitation studies with anti gag sera and the kinetics of cleavage of Gz-p65gag. Both Mo-MuLV Pr65gag and Gz p65gag were found to be very similar in primary sequence as judged by peptide mapping. P40 produced tryptic peptides that co-migrated with Mo-MuLV p30 peptides; P25 contained tryptic peptides that were also found in Mo-MuLV p15. Gz p30 and Gz-p15 contained the tryptic peptides of Mo-MuLV p30 and p15, respectively, that were found in P40 and P25. The Gz-p10 fraction contained a tryptic peptide that was also found in P40, but not p30. These results provide good evidence that the protease packaged within Mo-MuLV can cleave, in vitro, the gag-related polyprotein of Gz-MuSV in a manner very similar to the processing of Mo-MuLV Pr65gag in infected cell culture systems. PMID- 2981964 TI - DNA sequence of the major inverted repeat in the varicella-zoster virus genome. AB - The major inverted repeat of 7319.5 base pairs is present at an internal site in the varicella-zoster virus genome and at one terminus. A DNA sequence of 7747 base pairs containing the repeat was determined and analysed. The G + C content of the repeat is not uniform, and is significantly higher than that of adjacent unique regions. The repeat contains a G + C-rich reiterated sequence, an A + T rich sequence with the potential of forming a hairpin structure which may form part of an origin of DNA replication, and three open reading frames predicted to encode primary translation products with approximate molecular weights of 140 000, 30 000 and 20 000. The possibility is discussed that the expression of other open reading frames near the genome termini may depend upon genome conformation. PMID- 2981966 TI - Antigenic and biochemical analysis of gB of herpes simplex virus type 1 and type 2 and of cross-reacting glycoproteins induced by bovine mammillitis virus and equine herpesvirus type 1. AB - An antiserum was produced to the oligomeric form of glycoprotein B (gB) induced by herpes simplex virus type 1 (HSV-1) strain 17. This antiserum gave a single common precipitin line in agar gel immunodiffusion with HSV-1, HSV-2, bovine mammillitis virus (BMV) and equine herpesvirus type 1 (EHV-1). It also neutralized HSV-1, HSV-2 and BMV but not EHV-1. Absorption of the antiserum with excess HSV-2 or BMV antigen resulted in an HSV-1-specific neutralizing antiserum. In immunoprecipitation, two proteins, gB and pgB, were precipitated from HSV-1- and HSV-2-infected cells and at least three from BMV- and EHV-1-infected cells. Glycoprotein B and pgB of three HSV-1 and three HSV-2 strains and the corresponding antigenically related glycoproteins of BMV- and EHV-1-infected cells were labelled with 125I, digested with trypsin and the resulting peptides separated by two-dimensional thin-layer chromatography or high-pressure liquid chromatography. The resulting profiles were found to be almost identical, suggesting considerable structural conservation of the peptide backbone of the antigenically related glycoproteins of these four viruses. PMID- 2981965 TI - Immunochemical characterization of pyrimidine kinase induced by varicella-zoster virus. AB - Thymidine kinase (TK) induced by varicella-zoster virus (VZV) was precipitated with ammonium sulphate and purified by Sephadex G-150, QAE-Sephadex and Blue Sepharose column chromatographies. The purified TK fraction also contained deoxycytidine kinase (dCK) activity and a 35000 mol. wt. (35K) polypeptide as a major component. The TK and dCK activities were both neutralized by anti-VZV serum. Antiserum to an extract of cells infected with a bromodeoxyuridine (BUdR) resistant mutant virus contained no antibody to the viral TK and dCK activities or to the 35K polypeptide. Antiserum to the purified viral TK fraction was prepared and absorbed with a lysate of BUdR-resistant mutant virus-infected cells. The resulting absorbed antiserum (anti-vTK serum) neutralized the viral activities of both TK and dCK, and specifically immunoprecipitated a 35K polypeptide from the lysate of parental virus-infected cells, but did not immunoprecipitate any detectable polypeptide from cells infected with BUdR resistant mutant virus. Anti-vTK serum stained mainly the nuclei of cells infected with the parental virus strain, but did not stain those infected with BUdR-resistant mutant virus by an indirect fluorescent antibody test. These results suggest that the 35K polypeptide produced in VZV-infected cells is responsible for the viral TK and dCK activities, and that the TK and dCK are mainly localized in the nuclei of infected cells. PMID- 2981967 TI - Interferon gamma production by herpes simplex virus antigen-specific T cell clones from patients with recurrent herpes labialis. AB - Nineteen herpes simplex virus (HSV) antigen-specific human T lymphocyte clones were established from three volunteers with recent recurrent herpes labialis. All produced interferon gamma (IFN-gamma) at titres of 200 to 700 units/ml when cultured in vitro with HSV antigen and irradiated peripheral blood mononuclear cells (PBMC) as filler cells. All 10 of those clones whose phenotype was determined were Leu 4+, Leu 2-, Leu 3+. Interleukin 2 alone failed to induce IFN gamma in titres greater than 10 units/ml from these clones cultured at 10(4)/0.2 ml/well. However, the effect of different accessory or filler cells on IFN-gamma production by clones was quite marked. For example, high titres were produced when irradiated PBMC or plastic-adherent cells (predominantly monocytes) were added and low titres when macrophages and irradiated Epstein-Barr virus transformed B (EBV-B) cells were added. When tested for HSV antigen-stimulated IFN production alone, the irradiated PBMC and adherent cells produced low titres, but no detectable interferon was produced by the others. However, with higher concentrations of EBV-B cells, low concentrations of IFN-alpha were occasionally produced. Irradiation strikingly reduced IFN-alpha-production by PBMC. The IFN alpha and -gamma produced by accessory cells may contribute to total IFN production by priming the production by cloned cells, and acting in synergy with IFN-gamma produced by the cloned cells. Alternatively, the effect may be due to the presence of permissive concentrations of other lymphokines such as the interleukins. Interferon production by cloned T lymphocytes in the presence of non-producing macrophages was maximal within 24 h, much faster than with a similar polyclonal system, although attaining lower titres. EBV-B cells from only one of three patients supported antigen-specific lymphocyte activation. Almost all cells of the three cell lines expressed DR antigens, while DS/DC antigens were also expressed on nearly all cells of the antigen-presenting line and, at lower densities, on two-thirds of the cells of the other two lines. PMID- 2981968 TI - Increased mutation frequency after herpes simplex virus type 2 infection in non permissive XC cells. AB - The effect of herpes simplex virus type 2 (HSV-2) infection on the frequency of mutations at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus was studied in the non-permissive XC cell line. When the cells were infected with 20 to 800 p.f.u./cell, there was initially a lag in cell growth and cell death, but after 4 days there was no difference in growth rate between infected and control cultures. However, the mutation frequency, as determined by the number of 6-thioguanine-resistant colonies, was increased in infected cultures by factors ranging from 2.5 to 10.3. This effect was found to be dependent on the multiplicity of infection. The maximum effect was obtained between 20 and 100 p.f.u./cell while further increase in the amount of virus resulted in a drop in the yield of mutants. The optimum multiplicity of infection was a reproducible characteristic but was variable between viral stocks. When a number of mutant clones were examined they were found to have HGPRT activities ranging from undetectable to 6.9% of wild-type, indicating that the mutations were in the HGPRT gene. These results show that, in a non-productive infection, HSV-2 particles can increase the mutation frequency. The possible mechanisms by which this effect is brought about in the host genome are discussed. PMID- 2981969 TI - Factors involved in interferon-induced or cholera toxin-induced steroidogenesis in Y-1 mouse adrenal tumour cells. AB - In addition to its antiviral effect, interferon, at high concentrations, stimulates steroidogenesis and provokes cell rounding in Y-1 mouse adrenal tumour cells. This stimulation was inhibited by cytochalasin B and colchicine. In contrast, dibutyryl cAMP and cholera toxin, also able to induce steroid production and cell rounding, increased steroid production even in the presence of these cytoskeleton-disrupting agents. The initial trigger for interferon or cholera toxin thus probably involves a distinct receptor organization. However, since both inducers increased cAMP synthesis in this differentiated cell line, the further metabolic steps of ketosteroid production could be the same. PMID- 2981970 TI - Neurovirulence of the intertypic poliovirus recombinant v3/a1-25: characterization of strains isolated from the spinal cord of diseased monkeys and evaluation of the contribution of the 3' half of the genome. AB - A tsRNA- intertypic recombinant, v3/a1-25, which has the 5' and 3' halves of the genome derived from the neurovirulent type 3 poliovirus strain 452/62 3D and the attenuated type 1 poliovirus strain LSc-gr3, respectively, was previously shown to cause severe paralytic poliomyelitis after intracerebral inoculation of monkeys. To ascertain whether the illness was caused by the recombinant itself or by temperature-resistant trRNA+ mutants that might have arisen in the inoculated monkeys, five independent virus strains have been isolated from the spinal cord of the diseased animals. While two of these isolates exhibited RNA+ and RNA +/- phenotypes, respectively, the other three strains retained the parental RNA- character. Except for the RNA+ strain, the RNase T1 oligonucleotide maps of the genomes of all the isolates revealed only a minimal deviation from the parental pattern. These results were interpreted to mean that v3/a1-25 is intrinsically neurovirulent despite the presence of a tsRNA- mutation(s) in the 3' half of its genome. Nevertheless, this mutation, or other peculiarities of the 3' half of the recombinant genome, may somewhat alleviate the pathogenicity of the virus. This notion was inferred from the fact that, when used in a relatively small dose (about 10(3) p.f.u.), v3/a1-25 appeared to exhibit a lower level of neurovirulence compared to either the wild-type parent 452/62 3D, or a closely related intertypic recombinant having the genome 3' half derived from a neurovirulent trRNA +/- type 1 poliovirus strain. The problem of genetic determination of poliovirus neurovirulence and attenuation is briefly discussed. PMID- 2981973 TI - Growth of defective subacute sclerosing panencephalitis viruses in human neural cells and their neurovirulence in mice. AB - A defective subacute sclerosing panencephalitis (SSPE) virus which had been passaged in human embryonic lung cells was transferred to cultures of three neural cell types: neuroblastoma, oligodendroglioma and glioblastoma. The growth characteristics of the virus in these cells were essentially similar to those in non-neural cells. On the other hand, a marked difference in neurovirulence was noticed for the virus grown in neural cells when examined by intracerebral inoculation into mice. The virus passaged in neuroblastoma and oligodendroglioma cells showed high neurovirulence, inducing an acute encephalitis, whereas the virus passaged in human embryonic lung cells and that in glioblastoma cells did not show neurovirulence. These results suggest that the virus recovered its neurovirulence after passage in certain human neural cells. PMID- 2981971 TI - Sequence determination of the Sendai virus fusion protein gene. AB - From a genomic DNA library of Sendai virus, we have identified and sequenced clones corresponding to the F glycoprotein gene. The limits of the F gene region were defined by mapping the 5' and 3' ends of the mRNA with S1 nuclease. The Sendai virus F gene is 1821 nucleotides long. The predicted primary translation product of the single long open reading frame would code for a protein of 565 amino acids, containing a putative signal peptide, three carbohydrate addition sites, a hydrophobic region corresponding to the known cleavage/activation site of FO, and a long, very hydrophobic region near the C-terminus which probably represents the transmembrane region of the protein. The signal peptide cleavage site of the mature protein was determined by mass spectrometry. Interestingly, the amino acid sequence surrounding the cleavage/activation site of the Sendai virus F protein shows significant homology to the same region of the influenza B and C virus HA proteins, suggesting that these genes may have evolved from a common ancestor. The ability of the Sendai virus F protein to fuse membranes relative to its primary structure is discussed. PMID- 2981972 TI - The reversion of temperature-sensitive mutants of encephalomyocarditis virus upon serial passage in three continuous cell lines. AB - Several temperature-sensitive mutants of encephalomyocarditis virus were serially passed numerous times in HeLa, L929 and BHK-21 cells. The reversion of most of the mutants occurred most rapidly in BHK-21 cells and least rapidly in HeLa cells. PMID- 2981974 TI - Haemadsorption and haemagglutination by feline leukaemia viruses. AB - The interaction of feline leukaemia viruses (FeLV) with erythrocytes was investigated. Haemadsorption (HAd) was observed on the surface of feline embryonic fibroblast cells infected with FeLV. HAd was detected in various degrees when cat, hamster or horse erythrocytes were incubated with cells infected with viruses of subgroup C (FeLV-C) and on cells infected with some FeLV subgroup A viruses (FeLV-A), but not on cells infected with FeLV subgroup B viruses (FeLV-B). HAd of sheep erythrocytes was detected on cells infected with some FeLV-C viruses. The HAd of hamster erythrocytes on cells infected with FeLV C/Sarma virus was inhibited by antisera against gp70 or p15(E) but not by sera to the other FeLV structural polypeptides. HAd inhibition was also exhibited by cat sera which had FeLV-neutralizing activity but not by sera of specific pathogen free cats. Haemagglutination by FeLV-C viruses was demonstrated after the virus was treated with neuraminidase and phospholipase C, or Tween-80 and ether. Contrary to expectations from the pattern observed by HAd, all FeLV-A viruses had similar haemagglutinin (HA) activity to FeLV-C viruses. FeLV-B viruses did not possess an HA. PMID- 2981976 TI - The seroepidemiology of infection due to Epstein-Barr virus in southern India. AB - We investigated the seroepidemiology of infection due to Epstein-Barr virus (EBV) in 181 south Indian subjects aged 0-25 years using the indirect immunofluorescence method to titrate antibodies to viral capsid antigen (VCA), nuclear antigen (EBNA), and early antigen (EA). The age-specific prevalence of IgG antibodies to VCA rose rapidly to 90% by the age of 5 years. The prevalence of VCA-specific IgM and the geometric mean titre of VCA-specific IgG antibodies were highest between the ages of 6 months and 2 years, the median age of primary infection being 1.4 years. Thus primary EBV infection occurs early in life. EA antibody prevalence was highest (55%) in the third year of life and remained between 30% and 40% thereafter. This pattern of EA antibody prevalence suggests that the latent EBV infection that persists lifelong after primary infection may be reactivated in many individuals. EBNA antibody prevalence was low until the age of 2 years but rose to 80% in the fourth year. Geometric mean titres of antibodies to EA and EBNA were low and stable at all ages. These results are similar to data from areas where EBV-associated Burkitt's lymphoma is endemic and indicate a high EBV infection load early in life. PMID- 2981975 TI - Isolation of felid herpesvirus I from the trigeminal ganglia of latently infected cats. AB - Felid herpesvirus I (FHV I) is a respiratory pathogen of cats. Recovered animals remain latently infected but episodes of virus shedding may occur, particularly after stress. Previous attempts to identify the site(s) of latency of FHV I using co-culture, explant or tissue homogenization methods have been unsuccessful. Using a tissue fragment culture technique, however, FHV I was isolated from the trigeminal ganglia of three of 17 cats latently infected with field virus. No virus was found in four cats previously infected with a cold-adapted vaccine strain of FHV I, or in four cats given vaccine strain and field virus. The three trigeminal ganglion isolates had similar plaque size and morphology to the vaccine strain, the plaques being significantly smaller (P less than 0.001) than those of the field viruses. PMID- 2981977 TI - Study of herpes simplex virus type 1 populations obtained from recurrences and primary infections. AB - The analysis of 23 clinical isolates of herpes simplex virus type 1 (HSV-1) showed that 15 of 15 isolates that had undergone a few passages in tissue culture (fresh isolates) and two of eight isolates that had never been passaged (new isolates) were composed of a mixed population with respect to plaque morphology in Vero cells. Cloning and characterization of 10 large plaque viruses (L variants) and nine small plaque viruses (S variants), obtained from seven different isolates, showed the following. BamHI DNA restriction patterns of the L and the S variants from a single isolate differed only with respect to the electrophoretic mobility of the fragments that contain reiteration of specific sequences; they did not differ regarding the presence or the absence of restriction endonuclease cleavage sites. The L and S variants differed with respect to the electrophoretic profiles of infected cell glycoproteins, thermosensitivity of growth and plaquing efficiency at 39 degrees C, and, at least in the case of the two couples of variants that we tested, pathogenicity for the mouse. The hypothesis that the L variants might arise from the S variant during in vivo replication is discussed. PMID- 2981978 TI - Presence of antibodies to human lymphoma-leukemia virus (HTLV-I) in Germans with symptoms of the acquired immunodeficiency syndrome (AIDS). AB - Sera from German patients exhibiting symptoms compatible with the acquired immune deficiency syndrome (AIDS) or the lymphadenopathy syndrome (LAS) were assayed for antibodies against human T-cell lymphoma/leukemia virus (HTLV-I)-related antigens by enzyme immunoassay, indirect immunofluorescence, and radioimmunoprecipitation. Antibodies against HTLV were detected in 3 out of 31 sera. PMID- 2981980 TI - Attenuated Junin virus infection in Callithrix jacchus. AB - Twenty marmosets, male Callithrix jacchus, were used during this study. Fifteen of the marmosets were inoculated with 5,000 TCID50 of the attenuated XJC13 strain of Junin virus by intramuscular route and five were left as uninoculated controls. Animals were observed for a 420-day period. In order to carry out virologic, hematologic, serologic, and histologic studies the animals were bled and/or killed at different days post infection(pi). Results obtained showed that the attenuated strain produced an infection with no mortality or signs of illness. There was only a slight loss of weight at 18-40 days pi, which was soon recovered. Viremia was present from day 6 to 22, titers peaking at 4.0 log. Viral spread was limited to the lungs, spleen, lymph nodes, and bone marrow in the animal killed on day 14. No virus was found in the organs of the animal killed on day 23, and neither hematologic alterations nor pathologic lesions were seen in these monkeys except for ganglionar hypertrophy with immunoblast proliferation. Antigen was detected by immunofluorescence (IF) in lymph nodes, spleen, adrenals, lungs and brain. Neutralizing antibodies were detected from the third week onward. Protection conferred by the XJC13 strain proved effective when XJC13 inoculated monkeys were challenged with 1,000 TCID50 of the pathogenic XJ strain at days 60 or 380 pi, while normal controls died. When viral persistence was searched for on days 370, 390, and 420 pi, no infectious virus was detected, but viral antigen was seen in certain organs, which, however, lacked tissue damage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2981979 TI - Antiviral effect of flavonoids on human viruses. AB - The effect of several naturally occurring dietary flavonoids including quercetin, naringin, hesperetin, and catechin on the infectivity and replication of herpes simplex virus type 1 (HSV-1), polio-virus type 1, parainfluenza virus type 3 (Pf 3), and respiratory syncytial virus (RSV) was studied in vitro in cell culture monolayers employing the technique of viral plaque reduction. Quercetin caused a concentration-dependent reduction in the infectivity of each virus. In addition, it reduced intracellular replication of each virus when monolayers were infected and subsequently cultured in medium containing quercetin. Preincubation of tissue culture cell monolayers with quercetin did not affect the ability of the viruses to infect or replicate in the tissue culture monolayers. Hesperetin had no effect on infectivity but it reduced intracellular replication of each of the viruses. Catechin inhibited the infectivity but not the replication of RSV and HSV-1 and had negligible effects on the other viruses. Naringin had no effect on either the infectivity or the replication of any of the viruses studied. Thus, naturally occurring flavonoids possess a variable spectrum of antiviral activity against certain RNA (RSV, Pf-3, polio) and DNA (HSV-1) viruses acting to inhibit infectivity and/or replication. PMID- 2981981 TI - Extensive-stage small-cell bronchogenic carcinoma: intensive induction chemotherapy with high-dose cyclophosphamide plus high-dose etoposide. AB - Seventeen ambulatory patients with extensive-stage small-cell lung cancer received one or two courses of high-dose induction chemotherapy consisting of cyclophosphamide (100 mg/kg) plus etopside (1,200 mg/m2) followed by four or five cycles of conventional-dose cyclophosphamide (1,000 mg/m2), doxorubicin (40 mg/m2), and vincristine (1.4 mg/m2) (CAV) given every 21 days. No additional chemotherapy was given until relapse or progression was documented. Response was assessed initially after high-dose induction therapy and again after completion of all chemotherapy. After induction therapy, 16/17 (94%) patients had achieved an objective response, including five (29%) complete responses and 11 (65%) partial responses. Two partially responding patients improved to a complete response after CAV, while one partial responder progressed and one patient died of an intercurrent illness while receiving CAV. Thus, the overall response after completing all chemotherapy was 15/16 (94%), including seven (44%) complete responses and eight (50%) partial responses. Median response duration was six months (range, 3 to 11 months), and overall median survival was ten months (range, 2 to 17 months). All 31 courses of induction therapy were associated with leukopenia (less than 1,000/microL), 81% with thrombocytopenia (less than 20,000/microL), and 77% with fever (greater than 38.5 degrees C). Seven episodes of bacteremia and one axillary abscess were documented, and there was one treatment-related death (6%). These toxicities are similar to that produced by high-dose etoposide alone. High-dose cyclophosphamide combined with high-dose etoposide can be administered to ambulatory patients with extensive-stage small cell lung cancer without requiring bone marrow transplantation to reestablish hematopoiesis. Complete response and median survival rates, however, are similar to those obtained with less intensive therapy. PMID- 2981982 TI - Randomized trial of three combinations of cisplatin with vindesine and/or VP-16 213 in the treatment of advanced non-small-cell lung cancer. AB - One hundred sixty-seven evaluable patients with non-small-cell lung cancer were randomized to receive high-dose cisplatin and vindesine (PVD), or cisplatin and VP-16-213 (etoposide epipodophyllotoxin) (PVP), or cisplatin with VP-16-213 and vindesine (PVPVD). The patient distribution and characteristics were similar in all the treatment arms. The response rate differences (35% in PVD arm, 30% in PVP arm, and 22% in PVPVD arm) were not statistically significant (P = .33). Response durations were 43 weeks in the PVD arm, 20 weeks in the PVP arm, and 27 weeks in the PVPVD arm. Median survival was 29 weeks in the PVD and PVP arms and 28 weeks in the PVPVD arm. Median survival time of responding patients was 76 weeks in the PVD arm and 65 weeks in the PVP arm; 78% of patients were alive at 22+ to 87+ weeks follow-up in the PVPVD arm. Myelosuppression was similar in all three treatment arms. Significantly more azotemia occurred in the PVD arm than in the PVP and PVPVD arms (P = .002), and significantly more neuropathy in the PVD and PVPVD arms than in the PVP arm (P = .003 and .005). All the treatment arms have similar antitumor activity in non-small-cell lung cancer, but the PVP combination is slightly less toxic than the PVD and PVPVD treatment arms. PMID- 2981984 TI - Lung cancer in Hodgkin's disease: association with previous radiotherapy. AB - Seven cases of lung cancer were observed in patients with Hodgkin's disease (HD) since 1970. The risk ratio for the development of lung cancer among HD patients was 5.6 times that expected in the general population. The pertinent clinical data from these patients are described and compared to 28 additional patients reported from other institutions. Small-cell lung cancer represented the predominant histologic type of lung cancer encountered in both smoking and nonsmoking patients with HD, accounting for 42% of cases overall and greater than 55% of cases reported in reviews of second malignancies. Tobacco use was noted in only 53% of patients. Twenty-eight (94%) of 30 patients developing metachronous lung cancer received supradiaphragmatic irradiation as primary therapy for HD. Nineteen (68%) of these patients received subsequent chemotherapy salvage. The median age at diagnosis of HD and lung cancer was 39 and 45 years, respectively. The interval between diagnosis of HD and metachronous lung cancer averaged seven years but appeared to vary inversely with age. HD patients treated with supradiaphragmatic irradiation or combined modality therapy may be at increased risk for developing lung cancer. The high frequency of in-field malignancies that we observed and the prevalence of small-cell lung cancer in both smoking and nonsmoking patients suggests that chest irradiation may influence the development of metachronous lung cancer in these patients. The finding of a mean latent interval in excess of seven years emphasizes the need for close long-term observation. PMID- 2981983 TI - Late intensification in small-cell lung cancer: a phase I study of high doses of cyclophosphamide and etoposide with autologous bone marrow transplantation. AB - We conducted a late dose intensification pilot study of small-cell lung cancer (SCLC) treated with high doses of cyclophosphamide (200 mg/kg) and etoposide (1.0 to 3.5 g/m2), administered during a period of 48 hours, as well as autologous bone marrow infusion. We have been able to administer safely 3 g/m2 of etoposide with the autologous bone marrow infusion and 1.5 g/m2 without it. Limiting extrahematopoietic toxicity appeared to take the form of irreversible cardiac failure. Complete responses have been obtained with our regimen for late dose intensification, but the duration of the responses and the survival rates of the patients were poor. This suggests that late dose intensification in incomplete responders is not superior to the usually reported results obtained with standard regimens for the treatment of SCLC. PMID- 2981985 TI - Radionuclide venography in Budd-Chiari syndrome with intrahepatic vena-caval obstruction. AB - Radionuclide imaging of the inferior vena cava (RIVC) was performed by injecting [99mTc] phytate into a dorsal pedal vein, as an initial diagnostic procedure for eight patients with clinical features of Budd-Chiari syndrome. In five of them, membranous occlusion of the inferior vena cava (IVC) was proved by contrast venography and subsequent surgery. The other three patients, with histologically verified hepatocellular carcinoma, were proved to have tumor-induced narrowing or occlusion of the IVC by contrast venography. The RIVC findings include a sharply truncated inferior vena cava with marked hand-up of activity, extensive collaterals, and delayed visualization of the heart. Our results indicate that RIVC is as accurate and specific as contrast venography, by demonstrating the occlusion of the IVC and collateral circulation from the functional aspect. This simple and noninvasive method could therefore be used as a first-line test in patients with unexplained edema, ascites, superficial abdominal venous collaterals, and even in patients with hepatocellular carcinoma, for the detection of obstruction in the inferior vena cava. PMID- 2981986 TI - Effect of vitamin D3 on imaging of experimental myocardial infarcts with technetium-99m pyrophosphate: further studies of the role of calcium. AB - We previously found that a pulse dose of vitamin D3 increased [99mTc]PPi uptake by rat myocardial necrosis. Vitamin D3 raised serum and lesion [Ca] but not, we now report, lesion [Fe]. We now also report that D3 increased [99Tc]PPi uptake by myocardial infarcts (L) in dogs from 0.345 +/- 0.007% administered (kg) dose/g in controls to 0.703 +/- 0.089 in treated (p less than 0.025). Vitamin D3 decreased uptake by dog bone (B) as measured in rib and sternum, increasing L/B from 1.10 +/- 0.23 to 2.30 +/- 0.52 (p = 0.06) X (L) was positively, (p less than 0.005) and uptake by sternum was negatively (p less than 0.05) correlated with serum [Ca] and [P], respectively. Scintigrams graded by a "blinded" observer, showed 4+, 4+, and 3+ infarcts, respectively, in three D3-treated dogs, and 2+, 2+, and 1+, respectively, in three untreated. One untreated and one treated dog were negative; the latter showed the least response to D3 in serum [Ca] and [99mTc] in tissue samples. Vitamin D3 can increase L/B in dogs, enhancing scintigraphic images. PMID- 2981987 TI - Master's thesis policies in nursing education. AB - A telephone survey of 25 schools of nursing with doctoral programs was conducted to inquire about their master's thesis policies. Six programs require a thesis. Sixteen programs provide both thesis and other options, described in the paper. Other policy issues are discussed which include: relation of thesis to faculty research, criteria for serving on a master's thesis committee, and requirements for doctoral admission. Responses to the survey indicate a variety of types and degrees of research experience in master's programs. PMID- 2981988 TI - Microcomputers enhance student health fairs. AB - To stimulate student interest in learning about health, faculty assigned students the task of planning and implementing a university health fair. Two questions were posed: (1) Could sophomore students effectively plan and implement a health fair as a learning opportunity? (2) Could microcomputers be used appropriately to influence client participation and to enhance the nurse-client interactions? Literature regarding the efficacy of the health fair as a mode of health care delivery and as a student learning opportunity and literature addressing the advantages of microcomputer use were reviewed. Two health fairs were piloted. Microcomputers were employed in three areas: nutrition, stress, and substance abuse. Students, with faculty guidance, developed educational materials using principles of teaching, learning, and health promotion. Clients rated the health fair as congruent with their health needs. Students identified learning gained from the experience. Students and clients were receptive to computer use. Future directions are discussed. PMID- 2981989 TI - Academic inbreeding in nursing: intentional or inevitable? AB - Consensus exists that academic inbreeding inhibits innovation and scholarly achievement. Nevertheless, schools of nursing have been--and continue to be--more inbred than other schools within the university. In a follow-up survey to Miller and Ehnes' 1977 study, inbreeding has increased slightly--more junior than senior faculty are inbred, and rapidly expanding schools are the most inbred. A comparison of two schools (one highly inbred and one less so) demonstrates that schools do not recruit faculty from other schools in their region. The paper concludes that economic pressures may force deans to hire their own graduates (reducing recruitment costs) for junior positions and pay a relatively larger sum for a few senior level outbred faculty. Thus, academic inbreeding in nursing may be both inevitable and intentional. PMID- 2981990 TI - An experience in narrative writing to improve public health practice by students. AB - The experimental use of the Public Health journal is of value for several reasons. It demonstrates, again, that we may create more positive environments for learning in nursing. We can use techniques from various disciplines in order to get students to use more of their faculties and senses in thinking, problem solving, and communicating. The writing project arose out of the problem students have in assimilating Public Health values, in considering a wider range of alternatives for health care, and in feeling ill at ease with narrative writing and exploration. Since skills in cognition, thought, and writing are interrelated and affect each other, it is appropriate to apply writing methods and examine the outcomes in nursing students' work. The findings support the value of the narrative journal process. Although it does not operate alone in changing student overall response to Public Health in any given semester, it does affect who the students are and what their political-social values are. The students, in general, have been exposed to only traditional, conservative ideas regarding health care delivery and the state's social responsibilities. It is in experimenting with alternative ways to deal with the problems students have in approaching the Public Health model that we further aid them in their struggle to become professional nurse practitioners, who can effectively help people achieve "physical, mental, and social well being," health as defined by the World Health Organization. This is the objective we, as nursing faculty, need to keep before ourselves as a guiding principle at all times. PMID- 2981991 TI - Using a common health problem framework in a baccalaureate nursing curriculum. PMID- 2981992 TI - A learning experience in independent study. PMID- 2981993 TI - Creativity in clinical nursing. PMID- 2981994 TI - The use of assertiveness training in the clinical environment. PMID- 2981995 TI - The accreditation process as a learning experience. PMID- 2981996 TI - Curriculum design for educating nurse practitioners in adolescent health care. AB - In light of the limitation of available health care services for adolescents and of the small numbers of personnel to provide those services, it seems an opportune time for nurse practitioners to develop their role in adolescent health care. This paper describes the curriculum of the Adolescent Subspecialty Track which is a part of the Pediatric Nurse Practitioner program at the Yale University School of Nursing. One who graduates from the PNP program with a specialty in adolescent health care is able to practice as a PNP with an emphasis upon providing primary care to adolescents, to conduct clinical research, and to begin functioning as a leader in the profession. It is hoped that more nursing schools develop curricula for preparing nurse practitioners in the area of adolescent health care. PMID- 2981997 TI - Teaching power concepts. AB - Concepts and strategies presented here provide nurses with a new perspective from which to analyze and interact with power dynamics. Understanding fundamental concepts of power will help nurses enjoy a more equal status and bargaining position within the community of health professionals and in health care delivery systems. As nurses integrate and utilize this content for enhancing professional practices and client services, our public image will also continue to be strengthened. In so doing, our power base and sphere of influence will also be broadened. PMID- 2981998 TI - Application of computer technology in two colleges of nursing. PMID- 2981999 TI - Intestinal absorption and lipoprotein transport of (omega-3) eicosapentaenoic acid. AB - Adult male rats were surgically provided with a drainage catheter in the left thoracic lymphatic channel and an indwelling gastric catheter for constant infusion of saline-glucose. After an overnight fast, animals were given a single gastric dose of an aqueous emulsion containing one of the following 1-14C-labeled fatty acids: oleic, arachidonic or eicosapentaenoic acid, and [1,2 3H]cholesterol. Absorption was estimated by the appearance of radioactivity in lymph over a 24-h collection period, and the lymph lipoprotein distributions and lipoprotein lipid distribution were determined in the 24-h samples. Although there were apparent differences in the rates of eicosapentaenoic and arachidonic acid absorption, the overall appearance (after 24 h) of these acids in lymph was quantitatively equivalent to that of oleate. Furthermore, cholesterol absorption from each fatty acid medium was quantitatively similar. The distributions of each fatty acid among major lymph lipoproteins were similar with 93-95% recovered in chylomicrons and very low density lipoprotein (VLDL) fractions. Furthermore in these lipoproteins, 85-91% of each of the three fatty acids was recovered as triglycerides. With both polyunsaturated fatty acids, there was greater incorporation into phospholipids and their precursor partial glycerides than with oleate. These studies suggest that unesterified eicosapentaenoic acid is absorbed efficiently into the lymphatic system, and has lymph lipoprotein and lipid distribution comparable to those of the more typical mono- and polyunsaturated fatty acids. PMID- 2982000 TI - Effect of insulin administration on cardiac glycogen synthase and synthase phosphatase activity in rats fed diets high in protein, fat or carbohydrate. AB - Rats were fed a 70% carbohydrate, 70% protein, 70% fat, or a standard purified diet for 7 d to determine the effect of the diet on heart glycogen synthase response to an acute insulin challenge. Rats fed the high protein or the high fat diets, i.e., the carbohydrate-free diets, exhibited insulin resistance as evidenced by higher plasma glucose levels following insulin administration when compared to rats fed high carbohydrate or standard diet. The diets had no effect on the initial proportion of synthase in the active or I form. Insulin injection resulted in an increase in the proportion of synthase in the active form in rats fed the standard, high carbohydrate or high protein diets, but not in rats fed the high fat diet. Synthase phosphatase activity was similar in rats fed one of the four diets compared to rats fed a nonpurified diet. Thus the lack of synthase response to insulin in fat-fed rats was not due to diminished synthase phosphatase activity. Neither the diets nor insulin administration had any effect on the proportion of phosphorylase in the active or a form. Cardiac glycogen was significantly lower in rats fed the high fat diet than in those fed the standard diet. The latter was a surprising observation since the high fat diet was used to simulate a starved state and cardiac glycogen concentrations increase with starvation. PMID- 2982001 TI - A 10-year follow-up study of a group of workers exposed to isocyanates. AB - The ventilatory capacity of 68 workers who were exposed to isocyanates was determined. Forty-two of the workers had been studied in 1971. The overall ventilatory capacity of the subjects fell within the predicted range except that two subjects, both of whom were smokers, had minimal obstruction. No significant shift decrement was noted in any of the subjects. There was no evidence to suggest that an excess decrement in ventilatory capacity occurred during the period of observation; however, exposure to isocyanates had been well within the then-extant time-weighted average. PMID- 2982002 TI - Notification of a cohort of workers at risk of bladder cancer. AB - Surviving members of occupational cohorts studied in retrospective analyses of mortality usually are not notified individually of positive study results. However, concern has arisen that such results may contain risk information pertinent to study subjects. To evaluate the effects and implications of individually notifying survivors of such cohorts, we conducted a pilot notification study. Members of a cohort of 1,385 chemical workers who had handled carcinogenic amines were notified by mail that they were likely to be at increased risk of bladder cancer. Also a bladder cancer screening and follow-up program was established. The study demonstrated that notification is a complex action and that much care needs to be taken when communicating information on risk. Notification requires development of (1) criteria as to what constitutes a notifiable risk and (2) programs to meet the medical and social needs of the various parties involved in notification. PMID- 2982003 TI - TDI-induced oculorhinitis and bronchial asthma. AB - A case of allergic oculorhinitis induced by toluene diisocyanate (TDI) exposure in a subject who two years later developed bronchial asthma due to TDI is described. A 55-year-old nonatopic spray painter developed symptoms of oculorhinitis two or three hours after direct occupational exposure to polyurethane varnish; at the first examination neither specific nor nonspecific bronchial hyperresponsiveness was present. Two years later the patient, who had remained in his job, developed episodic dyspnea, wheezing, and cough immediately after TDI exposure, with persistence of oculorhinitis; at this time a slight immediate-type response to a specific bronchial provocative test with polyurethane varnish and TDI was observed. Nonspecific bronchial hyperresponsiveness was mild. Specific IgE to TDI-HSA conjugate was present at both the first and second examinations. We conclude that, in some cases, TDI may cause "allergic" oculorhinitis and bronchial asthma, probably with an immunological IgE-mediated mechanism. PMID- 2982004 TI - Evaluation of hydroxylapatite root implants in baboons. AB - To determine if there is a need for close conformation of a hydroxylapatite implant to the root socket for successful prevention of alveolar ridge resorption after tooth extraction, second premolars and lateral incisors of six mature baboons were extracted bilaterally, and custom-fitted root replicas or half length, plug-shaped forms of hydroxylapatite were implanted in the fresh sockets. Results showed that precise fit, as achieved by a custom-fitted implant, was not necessary for success. All implants were completely accepted, with evidence of new bone formation continuing for up to six months, the duration of the study. No resorption of the alveolar ridge was observed by gross or microscopic examination. PMID- 2982005 TI - Splenic reticuloendothelial function in children with cancer. AB - We studied splenic function in children with cancer by quantitation of pitted, or pocked, erythrocytes (pocked RBC count), that is, the percentage of erythrocytes containing one or more membrane-bound vesicles, as determined by phase interference microscopy. The mean pocked RBC count in 93 normal children and adults was 0.49% (range 0% to 2.0%), with only 2.4% of normal subjects having values greater than 1.5%. Mean pocked RBC count in 28 children after splenectomy was 37% (range 3.2% to 81%). Among 181 children with cancer (525 specimens), the mean pocked RBC count was 1.06% (range 0% to 12.6%). Fifty-nine (32%) patients had one or more values greater than 1.5%, and 25 (13.8%) children had measurements greater than 3.0%, a level previously suggested to have clinical significance. Elevated pocked RBC counts (greater than 1.5%) occurred in more than one third of children with Wilms tumor and acute lymphoblastic leukemia, and in both patients with juvenile chronic myelogenous leukemia. Elevations in pocked RBC counts were not related to specific chemotherapy regimens or to disease activity. Mild splenic reticuloendothelial hypofunction occurs in many children with cancer and may contribute to the risk of infection in these patients. PMID- 2982006 TI - Herpes zoster following varicella vaccine in a child with acute lymphocytic leukemia. PMID- 2982007 TI - Respiratory distress in the second week of life. PMID- 2982008 TI - Acquired immune deficiency syndrome in childhood. AB - The acquired immunodeficiency syndrome has been observed with increasing frequency in children with associated hemophilia, high-risk environmental backgrounds, and blood transfusions. AIDS should be considered in the differential diagnosis of childhood immunodeficiency, and it must be distinguished from congenital disorders. We emphasize the importance of epidemiologic, clinical, and laboratory data in diagnosis and aggressive management of infectious complications. The relationship between human retrovirus infection and AIDS remains to be precisely defined, especially with regard to cofactors that may play a role in the development of severe immunodeficiency following exposure to the agent. PMID- 2982010 TI - Role of alpha adrenoceptor activation in modulating the murine primary antibody response in vitro. AB - The present study shows that positive and negative modulation of the immunoglobulin M antibody response in mouse spleen cells immunized with sheep erythrocytes can be achieved by selective activation of alpha adrenoceptor subtypes. Alpha-1 adrenoceptor activation by methoxamine produced a number of spleen cells secreting immunoglobulin M antibody which was enhanced 63% above control on day 4 after immunization and which returned to control levels on days 5 (peak day of control antibody response), 6 and 7. This response mimicked the previously reported response produced by norepinephrine in the presence of propranolol, but not by norepinephrine alone. Alpha-2 adrenoceptor activation by clonidine produced no change when compared to control on days 4, 6 or 7, but produced a 50% suppression on day 5. Activation of both adrenoceptor subtypes by phenylephrine produced a control response on day 4, a depressed response on day 5 and an elevated response on days 6 and 7 by 50 and 64% above control, respectively. All drug responses were concentration-dependent and the methoxamine and clonidine responses were antagonized by phentolamine. These results suggest that antibody responses can be modulated by alpha-1 adrenoceptor activation to produce an enhanced response 1 day sooner than the peak control response and by alpha-2 adrenoceptor activation to produce a depressed response at the time of the peak control response. PMID- 2982009 TI - Indomethacin, prostaglandin E2 and transmission at the frog neuromuscular junction. AB - The effects of indomethacin on transmission at the frog neuromuscular junction were studied using intra- and extracellular voltage recordings. The drug had two dose-dependent effects on spontaneous and evoked quantal acetylcholine release. Low doses (less than or equal to 30 microM) irreversibly decreased and high doses (greater than or equal to 300 microM) reversibly increased release. Indomethacin also increased the latencies of evoked responses by increasing synaptic delays and usually increasing nerve action potential conduction times. Low doses are thought to inhibit cyclo-oxygenase specifically. To determine whether this single action might account for the presynaptic effects of the drug, other treatments that affect prostaglandin concentrations were also studied: 1) potassium 5,8,11,14-eicosatetraynoate (3 microM) and phenylbutazone (80 micron or 1 mM), two additional, molecularly different synthesis inhibitors; 2) prostaglandin E2 (30 nM-30 microM); and 3) sodium arachidonate (30 nM-30 microM), the essential fatty acid precursor of prostaglandin E2. Each treatment altered transmission, but perturbations in endogenous prostaglandin concentrations cannot account for all of their presynaptic effects. PMID- 2982011 TI - Neurochemical and functional correlates of naltrexone-induced opiate receptor up regulation. AB - The neurochemical and functional correlates of opioid receptor up-regulation after chronic antagonist administration in vivo and of down-regulation after withdrawal of antagonist were examined. Total brain opioid receptors increased 1.9-fold by day 8 of naltrexone administration, after which no further increase was observed; the newly synthesized or unmasked receptors exhibited an enhanced sensitivity to guanyl nucleotide modulation. Withdrawal from chronic naltrexone treatment resulted in a return to nearly control levels of receptor density and guanyl nucleotide sensitivity in a period of 6 days. These results suggest that up-regulation is accompanied by an increased coupling of the receptors to the inhibitory guanyl nucleotide binding protein (Ni) and that down-regulation involves the dissociation of the receptor/Ni complex. In experiments designed to target opiate receptor subtypes, long-term treatment with naltrexone was found to produce a coordinated up-regulation of brain mu and delta receptors, but did not cause a significant change in the density or affinity of kappa or sigma receptors. These findings indicate that the kappa and sigma opiate receptor classes may be subject to independent control mechanisms. Chronic naltrexone treatment also resulted in an enhanced morphine-induced analgesia. This result indicates that a functional supersensitivity occurs as a result of the selective up-regulation of mu and delta receptors. After withdrawal from naltrexone, supersensitivity to morphine-induced analgesia decreased monotonically and, in parallel to opioid receptor density, to prenaltrexone treatment levels within 6 days. Together, these results suggest a functional significance for antagonist induced mu and delta opiate receptor up-regulation. PMID- 2982013 TI - Pantoyl-gamma-aminobutyric acid facilitates cholinergic function in the central nervous system. AB - Pantoyl-gamma-aminobutyric acid (P-GABA) administered i.p. inhibited scopolamine- and atropine-induced locomotor activities in mice, but did not inhibit methamphetamine- and apomorphine-induced locomotor activities. In radiolabeled ligand binding experiments, P-GABA did not inhibit the bindings of [3H]quinuclidinyl benzilate, [3H]spiroperidol and [3H]apomorphine to rat brain membranes, but inhibited that of [3H]muscimol. GABA and P-GABA enhanced K+ (25 mM)-induced release of [3H]acetylcholine from slices of the cerebral cortex and hippocampus dose-dependently, and their effects were antagonized by bicuculline but not by tetrodotoxin. These results suggest that P-GABA binds to GABA receptors causing enhanced cholinergic neurotransmission in the central nervous system. The results are discussed in relation to the clinical use of P-GABA in treatment of Alzheimer's disease and senile dementia of the Alzheimer type. PMID- 2982012 TI - Chronic ethanol and (Na+-K+)-adenosine triphosphate: apparent adaptation in cation binding and enzyme conformation. AB - Sensitivity to ethanol, temperature dependence of apparent K+ affinity and temperature dependence of enzyme conformation of K+-p-nitrophenylphosphatase activity associated with (Na+,K+)-adenosine triphosphatase in brain membranes from rats treated chronically with liquid diets containing ethanol or isocaloric amounts of carbohydrate were compared. Three weeks' diet resulted in behavioral tolerance to ethanol. K+-p-nitrophenylphosphatase activity from tolerant rats was less sensitive than that from controls to inhibition by added ethanol, especially under conditions most favorable to ethanol inhibition. Apparent affinity for K+ was greater at all temperatures in ethanol-treated rats, and apparent delta S and delta H for K+-activation were reduced, opposite to effects of ethanol added in vitro. The amount of enzyme in the K+-sensitive conformation was greater at all temperatures in ethanol-treated rats, again opposite to effects of added ethanol. Apparent delta S and delta H of the conformational transition from E1 to E2 were reduced by ethanol treatment. These results are consistent with an apparent adaptation to chronic ethanol exposure in which membrane fluidity, at least in the neighborhood of (Na+,K+)-adenosine triphosphatase sites, is reduced, resulting in greater K+ affinity, more favorable transition to the phosphatase form of enzyme, a smaller entropy difference between native and K+-activated enzyme and reduced sensitivity to ethanol. PMID- 2982014 TI - Involvement of epinephrine in the presynaptic beta adrenoceptor mechanism of norepinephrine release from rat hypothalamic slices. AB - Using high-performance liquid chromatography with electro-chemical detector, we measured field impulse (5 or 2 Hz)- and high K+ (20 mM)-evoked release of endogenous norepinephrine from rat hypothalamic slices. Release by impulses at 5 Hz was tetrodotoxin-sensitive and both types of release were Ca++-dependent. Isoproterenol (10(-10) to 10(-8) M) dose-dependently facilitated impulse-evoked release and l-propranolol (10(-8) M) shifted dose-effect curve of isoproterenol to the right. Atenolol (10(-8) to 10(-6) M) or butoxamine (10(-9) to 10(-8) M), beta-1 and beta-2-antagonist, respectively, dose-dependently antagonized the facilitatory effect of isoproterenol (10(-8) M). Tazolol (10(-8) to 10(-7) M), beta-1-agonist, and salbutamol (10(-10) to 10(-8) M), beta-2-agonist, dose dependently increased impulse-evoked release. Epinephrine (10(-9) M) also facilitated impulse-evoked release and the action was antagonized by l propranolol (10(-8) M). Isoproterenol (10(-8) M) also facilitated high K+-evoked release in the presence of tetrodotoxin (3 X 10(-7) M) to exclude possible involvement of axonal conduction or neuronal loops. This facilitatory effect was antagonized by l-propranolol (10(-8) M). l-Propranolol (3 X 10(-7) M) alone decreased release by impulses at 2 Hz, but the d-isomer produced no effect. When rats were pretreated with 2,3-dichloro-alpha-methylbenzylamine, an inhibitor of phenylethanolamine N-methyltransferase, the enzyme catalyzing the formation of epinephrine from norepinephrine, 80 mg/kg i.p. before decapitation, the l propranolol-induced decrease was abolished completely.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982015 TI - Presynaptic opioid receptor subtypes in the rabbit ear artery. AB - In segments of rabbit ear arteries preincubated with [3H]noradrenaline, Leu enkephalin, D-Ala2-D-Leu-enkephalin and ethylketocyclazocine concentration dependently reduced the overflow of tritium and the vasoconstriction elicited by field stimulation (120 pulses every 14 min, 1 Hz, 0.3 msec pulse duration). The effects of Leu-enkephalin and ethylketocyclazocine were antagonized by naloxone which, given alone, increased the evoked overflow of tritium at the high concentration of 10 microM. Morphine failed to produce inhibition, and at 100 microM actually increased evoked 3H-overflow. Continued exposure to Leu enkephalin desensitized the tissue to this opioid; there was no cross desensitization to ethylketocyclazocine. In arteries not preincubated with [3H]noradrenaline, normorphine, fentanyl and morphiceptin did not change the vasoconstrictor response (5 pulses every min, 5 Hz, 0.3 msec pulse duration). Among various peptide agonists, Leu-enkephalin, D-Ala2-D-Leu-enkephalin and Met enkephalin were the most potent inhibitors. In a series of peptides with C terminal extensions of the Met-enkephalin chain, the potency decreased in the order Met-enkephalin greater than Met-enkephalin-Arg-Gly-Leu greater than Met enkephalin-Arg-Phe greater than BAM-12P greater than beta-endorphin. In a series of peptides with C-terminal extensions of the Leu-enkephalin chain, the potency decreased in the order Leu-enkephalin greater than dynorphin1-13 greater than dynorphin1-9 greater than alpha-neo-endorphin greater than dynorphin1-8 greater than dynorphin1-6 greater than dynorphin1-17. The delta-selective antagonist ICI 154129 counteracted the effect of Met-enkephalin but not that of dynorphin1-13, whereas naloxone counteracted the effect of either agonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982016 TI - Pharmacological evaluation of purified component and whole-cell pertussis vaccine in the cardiovascular system of rats. AB - The present study compares the cardiovascular activity of a newly introduced and possibly safer acellular pertussis vaccine with that of a conventional whole-cell pertussis vaccine. The vasodilation that occurs after beta-2 adrenoceptor stimulation with salbutamol, as well as the negative chronotropic action induced by the muscarinic receptor stimulant arecoline, were inhibited 4 days after vaccination with two combined diphtheria-pertussis-tetanus whole-cell vaccines and their plain pertussis product. Using doses equivalent to those given to humans, detoxified acellular pertussis component-combined vaccines (diphtheria pertussis-tetanus) and the nonabsorbed (detoxified pertussis component) induced minimal or no reduction in vascular beta-2 adrenergic and cardiac cholinergic receptor responsiveness as compared to saline-treated animals. Moreover, basal blood pressure values were significantly lower in whole-cell vaccine-treated rats than in animals vaccinated 4 days previously with acellular vaccine (or its ground products). Assuming that the present model can be used to predict the toxicity of pertussis vaccines in infants, it is concluded that the absence of a strong autonomic impairment might point toward the use of a less toxic acellular component vaccine in clinical practice. PMID- 2982018 TI - Pathogenicity of mouse hepatitis virus for preimplantation mouse embryos. AB - Mouse embryos which were hatched from the zona pellucida in vitro in the presence of mouse hepatitis virus (MHV) or outgrown on coverslips and then exposed to MHV were shown by immunohistochemical staining to have virally infected trophoblast cells. Zona-intact embryos incubated with MHV for 48 h (2-cell embryos) or 1.5 h (blastocysts) were resistant to infection. Morulae and early blastocysts collected from donor mice experimentally infected with MHV were not infected, but the medium in which they were flushed from the uterine horns was contaminated with virus. No virus was detected after embryos were washed through three changes of uncontaminated medium. MHV was transmitted to foster mothers when embryos were transferred in medium contaminated with the virus. Fetal and decidual tissues were not infected. We suggest that embryo transfer is an effective and simple alternative to Caesarian rederivation of MHV-contaminated mice. PMID- 2982017 TI - Localization of beta adrenoceptor subtypes in rat kidney by light microscopic autoradiography. AB - The characteristics and localization of beta adrenoceptor subtypes in rat kidney sections have been examined using [125I]cyanopindolol and in vitro labeling combined with autoradiography. Binding was stereoselective since the (-)-isomers of propranolol and pindolol were some two orders of magnitude more effective as competitors than the (+)-isomers. Competition curves obtained using the subtype selective antagonists ICI 118,551 (beta-2) and betaxolol (beta-1) had low pseudo Hill coefficients and were resolved into two distinct components representing beta-1 (63%) and beta-2 adrenoceptors (37%). Combined autoradiographic and histochemical studies using nuclear emulsion-coated coverslips and alkaline phosphatase staining showed that the majority of receptors were in the renal cortex and in the outer band of the medulla with fewer receptors associated with the inner medulla, papilla and renal blood vessels. Delineation of beta-1 and beta-2 adrenoceptor subtypes with the selective antagonists betaxolol and ICI 118,551 indicated that the highly localized receptors were predominately of the beta-1 subtype, associated with glomeruli and with tubules that from their staining characteristics and topography represent distal and cortical collecting tubules with few if any receptors associated with proximal tubules. Beta-2 adrenoceptors were more diffusely distributed in the cortex and there were minor areas of localization in the inner medulla. Although some glomerular beta adrenoceptors probably play a role in control of renin release, their distribution throughout this structure indicates that they also control other functions. The distribution of beta adrenoceptors in tubules corresponds well with the known distribution of beta adrenoceptor-stimulated adenylate cyclase in rat kidney and indicates that these receptors subserve a physiological function. PMID- 2982019 TI - Differential patterns in luteal prolactin and LH receptors during pregnancy in sows and ewes. AB - In the sow, a dramatic increase of LH specific binding was observed during the second half of pregnancy. This was due to an increase in receptor number (41 fmol and 95 fmol/mg protein at Days 50 and 105 respectively). The apparent association constant was unchanged. The pattern of prolactin receptor content showed two peaks at Day 60 and Day 105. Prolactin receptors increased earlier during pregnancy than did LH receptors, suggesting a possible role of prolactin in the induction of LH receptors. In the ewe, the receptor content of LH and prolactin did not change very much during pregnancy. The corpus luteum showed normal luteal function until parturition although it was not necessary for maintenance of pregnancy in the ewes. PMID- 2982020 TI - N-[2-hydroxy-5-[2-(methylamino)ethyl]phenyl]methanesulfonamide. A potent agonist which releases intracellular calcium by activation of alpha 1-adrenoceptors. AB - N-[2-Hydroxy-5-[2-(methylamino)ethyl]phenyl]methanesulfonamide (SK&F 102652) has been prepared and characterized pharmacologically. It is a potent agonist with an EC50 of 25 nM at alpha 1-adrenoceptors as determined in the isolated perfused rabbit ear artery. On the presynaptic alpha 2-adrenoceptors of the guinea pig atrium it was considerably weaker, demonstrating an EC50 for inhibition of neurotransmission of 1200 nM and thus an overall alpha 1/alpha 2 selectivity ratio of greater than or equal to 48. In the vascular smooth muscle of the canine saphenous vein an EC100 concentration of this agonist in the presence of zero external Ca2+ induced 37.9 +/- 1.4% of the maximal contractile response due to this agent while the endogenous ligand norepinephrine evoked only 14.5 +/- 0.4% of the maximum. In the presence of low (1 microM) external calcium, this agent produced 78.3 +/- 5.3% of maximum while norepinephrine gave 45.3 +/- 7.4%. This agent produces alpha 1-adrenoceptor-mediated contraction primarily by release of intracellular Ca2+ and should provide a useful tool for characterizing alpha 1 receptor subtypes. PMID- 2982021 TI - Pharmacological basis of therapeutics: vascular alpha-adrenoceptors. PMID- 2982022 TI - Feline panleukopenia virus replicates in cells in which cellular DNA synthesis is blocked. AB - The components of the cell cycle for a feline embryo cell line were defined. Thymidine (6mM)-supplemented medium reversibly arrested cells 1 h into the S phase of the cell cycle and was used in a double blocking procedure to synchronize cells to the early S phase. The kinetics of feline panleukopenia virus replication in synchronized cells was studied by using (i) inclusion body formation, (ii) a plaque assay for cell-associated and cell-free virus under one step growth conditions, (iii) an enzyme immunoassay for viral protein, (iv) electron microscopy of infected cells, and (v) the detection and identification of viral replicative form DNA by restriction endonuclease analysis. Parallel studies by each of these procedures of the replication of feline panleukopenia virus in cells in which a 6 mM thymidine block was maintained indicated that parvovirus replicated with essentially similar kinetics in both unblocked, synchronized cells and in cells in which the block was maintained. Accordingly, a 6 mM thymidine-supplemented medium, although it effectively blocks cellular DNA synthesis, does not block the replication of parvovirus. PMID- 2982023 TI - A physical domain of herpes simplex virus ICP8 is expressed and active in Escherichia coli. AB - In this report, we describe a series of procedures to assay the function of fusion genes in Escherichia coli and the specific application to the carboxy terminal third of the herpes simplex virus type 1 (HSV-1) DNA-binding protein ICP8. E. coli cells containing the cloned HSV-1 BamHI G fragment with the HSV-1 BamHI-G-V site, map unit 0.388, nearest the tet promoter in pBR322 synthesized an active product containing a portion of ICP8. The new product induced phenotypic alterations in recipient hosts that were measurable and stable yet limited to the stability of the plasmid. The corresponding cloned DNA from the characterized HSV 1 DNA-binding protein mutant tsHA1 exhibited a predictable temperature-sensitive phenotype. Screening procedures based on the loss of induction of the parental plasmid-induced phenotype in E. coli cells allowed us to select additional mutations. One of these, which conferred a phenotype different from that of tsHA1, was transferred to the viral genome by marker transfer techniques. We suggest that any mutant could be isolated in any sequence, provided that the wild type coding sequences induce alterations in E. coli cells. The observed alterations should have relevance in determining the mode of action of the protein in its normal environment. PMID- 2982024 TI - Plasma membrane orientation of simian virus 40 T antigen in three transformed cell lines mapped with monoclonal antibodies. AB - Simian virus 40 large T antigen transforms cells from several species. Recent studies show that it is present on the cell surface. As in other tumor virus systems, this may be important for transformation. We have used a radioimmunoassay to map antigenic determinants on living and formaldehyde-fixed transformed cells with six different monoclonal antibodies to T antigen. Nonrelevant monoclonal antibodies of the same subclasses served as controls. With the transformed mouse line SVT2, antibody PAb 101, which reacts with the C terminal region of T antigen, and PAb 1700, which is directed against an internal region of T, reacted with both formaldehyde-fixed and living cells. Antibodies PAb 402 (C terminus) and 419 (N terminus) reacted only with living cells, their determinants being destroyed upon formaldehyde fixation. Antibodies PAb 405 (C terminus) and 100 (internal) fail to react on either fixed or living cells. Similar results were obtained on the simian virus 40-transformed human line SV80 and the fixed hamster line CHLwt23, although all antibodies failed to react with living CHLwt23 cells. The data suggest that T antigen is inserted into the plasma membrane of transformed cells in a specific, nonrandom manner, with the C and N termini exposed on the cell surface and the midportion either buried in the lipid bilayer, hidden by the tertiary structure of T antigen, or masked by a post translational modification such as fatty acid acylation. PMID- 2982025 TI - Vesicular stomatitis virus-infected cells fuse when the intracellular pool of functional M protein is reduced in the presence of G protein. AB - Five highly cytolytic strains of both Indiana and New Jersey serotypes of vesicular stomatitis virus were shown to induce cell fusion in BHK-21 and R(B77) cells. Inhibition of protein synthesis after the eclipse period of viral replication is a prerequisite for vesicular stomatitis virus-induced cell fusion. Pulse-chase experiments showed that inhibition of protein synthesis would lead to a drastic reduction in the intracellular pool of M protein as compared with other proteins. A temperature-sensitive mutant defective in M protein function (G31) was the only mutant of the five complementation groups to spontaneously induce polykaryocytes at the nonpermissive temperature. Previously, G protein has been shown to play a role in vesicular stomatitis virus-induced cell fusion. These results suggest that the combination of the presence of G protein on the virus infected cell surface and the absence of functional M protein or a reduced level of intracellular M protein promotes cell fusion. On the basis of this study, we propose that vesicular stomatitis virus infection can induce cell fusion when the functional M protein pool declines to a critical level while G protein remains on the cell surface. PMID- 2982026 TI - Identification of an enveloped phage, mycoplasma virus L172, that contains a 14 kilobase single-stranded DNA genome. AB - We have found that mycoplasma virus L172 is an enveloped globular virion containing circular, single-stranded DNA of 14.0 kilobases. L172 has been reported by other workers to have a double-stranded DNA genome of 13 to 17 kilobase pairs and has been classified as a plasmavirus, a group for which mycoplasma virus L2 is the type member. Mycoplasma viruses L172 and L2 differ in genome size and structure, DNA base composition, and protein composition, and they have no detectable DNA homology. As the only reported enveloped virion containing single-stranded DNA, L172 represents a new group of viruses. PMID- 2982027 TI - Reovirus serotype 1 intestinal infection: a novel replicative cycle with ileal disease. AB - After oral inoculation, reovirus serotype 1 strain Lang was shown to specifically infect the epithelial cells of the ileum, while sparing the epithelial cells in the duodenum, jejunum, and colon. The initial site of replication was localized in cells of the crypts of Lieberkuhn adjacent to Peyer's patches. Virus was subsequently found by immunoperoxidase staining in cells migrating up the crypt villus complex throughout the ileum. The severity of the pathological changes in the ileum was proportional to the concentration of the viral inoculum. This site specific infection of the ileum by reovirus may provide a model for diseases that are restricted to specific sites in the intestine. PMID- 2982028 TI - A cis-acting sequence promotes removal of simian virus 40 DNA from the replication pool. AB - Pulse-labeled simian virus 40 (SV40) DNA is removed from the pool of molecules available for replication (i.e., it ceases to reenter replication) a few hours after synthesis. We studied this cessation of reentry with mutants containing different deletions in the structural genes of SV40. The DNAs of two independent deletion mutants, dl-1007 (24% deletion) and dl-1003 (8% deletion), were used as templates for further DNA synthesis (i.e., they reentered replication) to a greater extent than was wild-type DNA. The alteration in reentry kinetics was not because the DNAs were smaller; other deletion mutations that were from 76 to 85% of the length of wild-type DNA (dl-BE and dl-1133 with a deletion in the late region and F8dl with a deletion in the early region) did not reenter replication to a greater extent than the wild type did. Cotransfection experiments showed that the mutant phenotypes of dl-1007 and dl-1003 were poorly complemented, if at all, by the wild type. Thus, we propose that there is a cis-acting sequence located in the HindIII E fragment of SV40, not present in either of these mutants, that promotes the efficient removal of DNA from the replication pathway. PMID- 2982030 TI - Inverse relationship between constitutive gamma interferon production and human T cell lymphoma/leukemia virus expression in cultured T lymphocytes. AB - Particular interest in human T lymphocyte lymphoma/leukemia virus (HTLV) derives from the close association of HTLV with several types of human mature T lymphocyte malignancies and the strong possibility that HTLV is the causative agent of this group of leukemias and lymphomas. This is the first report to show that HTLV expression in T lymphocytes cultured in vitro is inversely proportional to constitutive gamma interferon production. Of 16 fresh T lymphocyte cultures established from patients with mature T lymphocyte neoplasias, 3 were grown continuously for over 3 years and 13 were grown for 2 to 8 months in culture. Of the 16 cultures, 9 were HTLVp19 positive and interferon negative, whereas the remaining 7 were HTLVp19 negative or weakly positive and also interferon positive (12 to 105 U/ml). The prototype HTLV-positive T-cell line (HUT102) was examined over a long-term culture and after selective cell cloning for high virus yield. Results indicate that early-passage, low-HTLV-producing HUT102 cells constitutively produced significant levels of gamma-immune interferon. In late passage and cloned HUT102 cells, an increase in HTLV production was concordant with a decrease in constitutive interferon production and the loss of mature T lymphocyte antigens. Transformation of human umbilical cord blood lymphocytes by HTLV was possible only after cocultivation with the non-interferon, high virus producing, cloned HUT102 T lymphocytes. The inverse relationship between interferon and HTLV production was also observed when normal human umbilical cord blood and adult T lymphocytes were transformed by HTLV and maintained in culture. PMID- 2982031 TI - Novel rearrangements of herpes simplex virus DNA sequences resulting from duplication of a sequence within the unique region of the L component. AB - We constructed insertion mutants of herpes simplex virus type 1 that contained a duplication of DNA sequences from the BamHI-L fragment (map units 0.706 to 0.744), which is located in the unique region of the L component (UL) of the herpes simplex virus type 1 genome. The second copy of the BamHI-L sequence was inserted in inverted orientation into the viral thymidine kinase gene (map units 0.30 to 0.32), also located within UL. A significant fraction of the progeny produced by these insertion mutants had genomes with rearranged DNA sequences, presumably resulting from intramolecular or intermolecular recombination between the BamHI-L sequences at the two different genomic locations. The rearranged genomes either had an inversion of the DNA sequence flanked by the duplication or were recombinant molecules in which different regions of the genome had been duplicated and deleted. Genomic rearrangements similar to those described here have been reported previously but only for herpes simplex virus insertion mutants containing an extra copy of the repetitive a sequence. Such rearrangements have not been reported for insertion mutants that contain duplications of herpes simplex virus DNA sequences from largely unique regions of the genome. The implications of these results are discussed. PMID- 2982029 TI - Two separable functional domains of simian virus 40 large T antigen: carboxyl terminal region of simian virus 40 large T antigen is required for efficient capsid protein synthesis. AB - The carboxyl-terminal portion of simian virus 40 large T antigen is essential for productive infection of CV-1 and CV-1p green monkey kidney cells. Mutant dlA2459, lacking 14 base pairs at 0.193 map units, was positive for viral DNA replication, but unable to form plaques in CV-1p cells (J. Tornow and C.N. Cole, J. Virol. 47:487-494, 1983). In this report, the defect of dlA2459 is further defined. Simian virus 40 late mRNAs were transcribed, polyadenylated, spliced, and transported in dlA2459-infected cells, but the level of capsid proteins produced in infected CV-1 green monkey kidney cells was extremely low. dlA2459 large T antigen lacks those residues known to be required for adenovirus helper function, and the block to productive infection by dlA2459 occurs at the same stage of infection as the block to productive adenovirus infection of CV-1 cells. These results suggest that the adenovirus helper function is required for productive infection by simian virus 40. Mutant dlA2459 was able to grow on the Vero and BSC 1 lines of African green monkey kidney cells. Additional mutants affecting the carboxyl-terminal portion of large T were prepared. Mutant inv2408 contains an inversion of the DNA between the BamHI and BclI sites (0.144 to 0.189 map units). This inversion causes transposition of the carboxyl-terminal 26 amino acids of large T antigen and the carboxyl-terminal 18 amino acids of VP1. This mutant was viable, even though the essential information absent from dlA2459 large T antigen has been transferred to the carboxyl terminus of VP1 of inv2408. The VP1 polypeptide carrying this carboxyl-terminal portion of large T could overcome the defect of dlA2459. This indicates that the carboxyl terminus of large T antigen is a separate and separable functional domain. PMID- 2982033 TI - Physical locus of the DNA polymerase gene and genetic maps of bacteriophage T5 mutants. AB - Segments of DNA that contained the DNA polymerase gene of bacteriophage T5 were isolated. The physical locus of the gene was identified by transforming Escherichia coli with purified DNA fragments generated by restriction enzyme digestions, and the transformed cells were used to rescue amber mutants of T5 with mutations in the gene for DNA polymerase. The method is applicable to any other gene that has mutations with low reversion frequencies. We studied the following mutations of the T5 DNA polymerase gene, reading from left to right by the standard convention (D. J. McCorquodale, Crit. Rev. Microbiol. 4:101-159, 1975): D7, D8, aml, ts5E-ts53, am6, and D9. These loci were found to reside within three pieces of DNA with a total length of 3,600 base pairs. Because the structural gene for T5 DNA polymerase is estimated to be 2,600 base pairs long, the whole structural gene may reside in these segments. These are located 58.3 to 61.3% of the distance from the left end of the DNA. The left-end piece of the DNA (1,100 base pairs) containing the polymerase gene has loci D7 and D8, and the right-end piece (1,600 base pairs) has locus D9, according to the results of the transformation assay. These results are consistent with the genetic map. PMID- 2982032 TI - Sensitivity of arabinosyladenine-resistant mutants of herpes simplex virus to other antiviral drugs and mapping of drug hypersensitivity mutations to the DNA polymerase locus. AB - Seven herpes simplex virus mutants which have been previously shown to be resistant to arabinosyladenine were examined for their sensitivities to four types of antiviral drugs. These drugs were a pyrophosphate analog, four nucleoside analogs altered in their sugar moieties, two nucleoside analogs altered in their base moieties, and one altered in both. The seven mutants exhibited five distinct phenotypes based on their sensitivities to the drugs relative to wild-type strain KOS. All mutants exhibited resistance to acyclovir and arabinosylthymine, as well as marginal resistance to iododeoxyuridine, whereas all but one exhibited resistance to phosphonoformic acid. The mutants exhibited either sensitivity or hypersensitivity to other drugs tested--2'-nor deoxyguanosine, 5-methyl-2'-fluoroarauracil, 5-iodo-2'-fluoroarauracil, and bromovinyldeoxyuridine--some of which differed only slightly from drugs to which the mutants were resistant. These results suggest ways to detect and treat arabinosyladenine-resistant isolates in the clinic. Antiviral hypersensitivity was a common phenotype. Mutations conferring hypersensitivity to 2'-nor deoxyguanosine in mutant PAAr5 and to bromovinyldeoxyridine in mutant tsD9 were mapped to nonoverlapping regions of 1.1 and 0.8 kilobase pairs, respectively, within the herpes simplex virus DNA polymerase locus. Thus, viral DNA polymerase mediates sensitivity to these two drugs. However, we could not confirm reports of mutations in the DNA polymerase locus conferring resistance to these two drugs. All of the mutants exhibited altered sensitivity to two or more types of drugs, suggesting that single mutations affect recognition of the base, sugar, and triphosphate moieties of nucleoside triphosphates by viral polymerase. PMID- 2982034 TI - Transcriptional activity of avian retroviral long terminal repeats directly correlates with enhancer activity. AB - Retroviral long terminal repeats (LTRs) contain elements responsible for the control of proviral transcription and gene expression. Molecular clones of the LTR region of a number of avian retroviruses have been isolated, and DNA sequence analysis of these clones reveals the existence of a related, but heterogeneous, family of LTRs. To examine the functional significance of the observed sequence differences, we have directly tested the abilities of several different avian retrovirus LTRs to act as promoters and enhancers of mRNA transcription. Our results indicate that large differences in LTR transcriptional activity exist and that these differences in gene expression directly correlate with LTR enhancer activity. In particular, we show that the LTR of Fujinami sarcoma virus is intermediate in both transcriptional and enhancer activity when compared with the very active LTRs of the exogenous viruses RAV-2 and Schmidt-Ruppin B and the much less active LTRs of the endogenous virus RAV-0 and its provirus ev-2. These results suggest that LTR enhancer activity may be the primary determinant of avian retroviral LTR transcriptional activity and, hence, oncogenic potential. PMID- 2982035 TI - Two related but differentially expressed potential membrane proteins encoded by the EcoRI Dhet region of Epstein-Barr virus B95-8. AB - Three mRNAs in the EcoRI Dhet region of Epstein-Barr virus B95-8 were mapped. Their 3' ends are coterminal. A latent gene containing three exons is transcribed from the ED-L1 promoter and was predicted to lead to expression of a 42 kilodalton protein. An unspliced late mRNA is produced by transcription from the ED-L1A promoter within the first intron of the above gene and was predicted to lead to expression of a 28-kilodalton protein corresponding to the C-terminal two thirds of the 42-kilodalton protein. Both proteins would have hydrophobic N terminal domains and highly acidic C-terminal domains and were predicted to span the cell membrane. An early promoter (ED-L2) is located in the 3' untranslated region of the above genes and was predicted to lead to expression of a 6.5 kilodalton polypeptide containing a C-terminal hydrophobic region. PMID- 2982036 TI - Characterization of the gene encoding herpes simplex virus type 2 glycoprotein C and comparison with the type 1 counterpart. AB - The gene encoding the glycoprotein C (gC) of herpes simplex virus type 1 maps to the region of the viral genome from 0.62 to 0.64. Recently, a herpes simplex virus type 2 glycoprotein previously designated gF and now designated gC was mapped to a homologous location. Analysis of the herpes simplex virus type 2 mRNA species encoded in this region revealed a major transcript of 2.5 kilobases, a 0.73-kilobase transcript (the 5' ends of which were mapped by primer extension), and several minor species, all nearly identical to the herpes simplex virus type 1 pattern. A polypeptide of ca. 60,000 daltons was identified by in vitro translation of hybrid-selected mRNA. A smaller protein of ca. 20,000 daltons was also mapped to this region. The nucleotide sequence of a 3.4-kilobase segment of DNA encompassing gC was determined, and an open reading frame of 1,440 nucleotides specifying a 480-amino acid protein with properties consistent with that of a glycoprotein was identified. Comparative DNA sequence analysis showed regions of limited homology within the coding sequences for gC and a deletion which results in 31 fewer amino acids in the gC-2 near the amino terminus of the protein. The carboxy termini of gC-1 and gC-2 are very similar, as are the 20,000 dalton proteins. PMID- 2982037 TI - A noninverting genome of a viable herpes simplex virus 1: presence of head-to tail linkages in packaged genomes and requirements for circularization after infection. AB - The wild-type herpes simplex virus 1 genome consists of two components, L and S, which invert relative to each other, giving rise to four isomers. Previously we reported the construction of a herpes simplex virus 1 genome, HSV-1(F)I358, from which 15 kilobase pairs of DNA spanning the junction between L and S components were deleted and which no longer inverted (Poffenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2690-2694, 1983). Further studies on the structure of HSV-1(F)I358 revealed the presence of two submolar populations among packaged DNA. The first, comprising no more than 10% of total packaged DNA, consisted of defective genomes with a subunit size of 36 kilobase pairs. The results suggest that this population arose by recombination through a directly repeated sequence inserted in place of the deleted L-S junction. The second minor population consisted of HSV-1(F)I358 DNA linked head-to-tail. Analyses of the structure of HSV-1(F)I358 DNA after infection indicated that the fraction of total DNA linked head-to-tail increased to approximately 40 to 50% within 30 min after exposure of cells to virus. The formation of head-to-tail linkages did not require de novo protein synthesis. Our interpretation of the results is that the termini of full-length DNA molecules are held together during packaging, that a small fraction of the termini is covalently linked during or after packaging, and that the remainder is covalently joined after the release of viral DNA from the infecting virus by either host or viral factors introduced into the cell during infection. PMID- 2982038 TI - Isolation of novel herpes simplex virus type 1 derivatives with tandem duplications of DNA sequences encoding immediate-early mRNA-5 and an origin of replication. AB - Two naturally occurring variations of herpes simplex virus type 1 (Patton strain) with novel tandem DNA sequence duplications in the S component were isolated, and the DNA was characterized. These variants were identified among a number of plaque isolates by the appearance of new restriction enzyme fragments that hybridized with radiolabeled DNA from the BamHI Z fragment (map coordinates 0.936 to 0.949) located in the unique S region. One isolate, SP26-3, carried a 3.1 kilobase-pair duplication defined by recombination between a site in the BamHI Z fragment and a site near the origin of replication in the inverted repeat sequence of the S component carried by the EcoRI H fragment. The other isolate, SP22-4, carried a 3.5-kilobase-pair duplication defined by a recombination event between a tandem repeat array in the BamHI Z fragment and a site near the amino terminus of the Vmw175 gene in the S-region inverted repeat sequence contained in the EcoRI K fragment. Both duplicated segments contained the entire immediate early mRNA-5 coding region as well as the origin of replication located in the inverted repeat sequence of the S component. The DNA sequence of each duplication joint was determined. PMID- 2982039 TI - Temperature-sensitive viral RNA expression in Moloney murine sarcoma virus ts110 infected cells. AB - We examined the mos-specific intracellular RNA species in 6m2 cells, an NRK cell line nonproductively infected with the ts110 mutant of Moloney murine sarcoma virus. These cells present a normal phenotype at 39 degrees C and a transformed phenotype at 28 or 33 degrees C, expressing two viral proteins, termed P85gag-mos and P58gag, at 28 to 33 degrees C, whereas only P58gag is expressed at 39 degrees C. It has been previously shown that 6m2 cells contain two virus-specific RNA species, a 4.0-kilobase (kb) RNA coding for P58gag and a 3.5-kb RNA coding for P85gag-mos. Using both Northern blot and S1 nuclease analyses, we show here that the 3.5-kb RNA is the predominant viral RNA species in 6m2 cells grown at 28 degrees C, whereas only the 4.0-kb RNA is detected at 39 degrees C. During temperature shift experiments, the 3.5-kb RNA species disappears after a shift from 28 to 39 degrees C and is detected again after a shift back from 39 to 28 degrees C. By Southern blot analysis, we have detected only one ts110 proviral DNA in the 6m2 genome. This observation, as well as previously published heteroduplex and S1 nuclease analyses which showed that the 3.5-kb RNA species lacks about 430 bases found at the gag gene-mos gene junction in the 4.0-kb RNA, suggests that the 3.5-kb RNA is a splicing product of the 4.0-kb RNA. The absence of the 3.5-kb RNA when 6m2 cells are grown at 39 degrees C indicates that the splicing reaction is thermosensitive. The splicing defect of the ts110 Moloney murine sarcoma virus viral RNA in 6m2 cells cannot be complemented by acute Moloney murine leukemia virus superinfection, since no 3.5-kb ts110 RNA was detected in acutely superinfected 6m2 cells maintained at 39 degrees C. The spliced Moloney murine leukemia virus env mRNA, however, is found in acutely infected cells maintained at 39 degrees C, suggesting that the lack of ts110 viral RNA splicing at 39 degrees C is not due to an obvious host defect. In sharp contrast, however, 6m2 cells chronically superinfected with Moloney murine leukemia virus produce a 3.5-kb RNA species at 39 degrees C as well as at 28 degrees C and contain proviral DNAs corresponding to the two viral RNA species.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982040 TI - Murine sarcoma virus ts110 RNA transcripts: origin from a single proviral DNA and sequence of the gag-mos junctions in both the precursor and spliced viral RNAs. AB - Our previous studies have argued persuasively that in murine sarcoma virus ts110 (MuSVts110) the gag and mos genes are fused out of frame due to a approximately 1.5-kilobase (kb) deletion of wild-type murine sarcoma virus 349 (MuSV-349) viral information. As a consequence of this deletion, infected cells grown at 39 degrees C appear morphologically normal, producing a 4-kb viral RNA and a truncated gag gene product, P58gag. At 33 degrees C, however, MuSVts110-infected cells appear transformed, producing two viral RNAs, about 4 and 3.5 kb in length, and two viral proteins, P58gag and P85gag-mos. Recent S1 nuclease analyses (Nash et al., J. Virol. 50:478-488, 1984) suggested strongly that at 33 degrees C about 430 bases surrounding the out-of-frame gag-mos junction and bounded by consensus splice donor and acceptor sites are excised from the 4-kb RNA to form the 3.5-kb RNA. As a result of this apparent splicing event, the gag and mos genes seemed to be fused in frame and allowed the translation of P85gag-mos. In the present study, DNA primers hybridizing to the MuSVts110 4- and 3.5-kb RNAs just downstream of the gag-mos junction points were used to sequence these junctions by the primer extension method. We observed that, relative to wild-type MuSV-349 5.2-kb RNA, the MuSVts110 4-kb RNA had suffered a 1,488-base deletion as a result of the fusion of wild-type gag gene nucleotide 2404 to wild-type mos gene nucleotide 3892. This gag-mos junction is out of frame, containing both TAG and TGA termination codons in the reading frame 42 and 50 bases downstream of the gag mos junction, respectively. Thus, the MuSVts110 4-kb RNA can only be translated into a truncated gag precursor containing an additional C-terminal 14 amino acid residues derived from an alternate mos gene reading frame. Similar analyses of the MuSVts110 3.5-kb RNA showed a further loss of both gag and mos sequences over those deleted in the original 1,488-base deletion. In the MuSVts110 3.5-kb RNA, we found that gag nucleotide 2017 was fused to mos nucleotide 3936 (nucleotide 2449 in the MuSVts110 4-kb genome). This 431-base excised fragment is bounded exactly by in-frame consensus splice donor and acceptor sequences. As a consequence of this splice event, the TAG codon is excised and the restoration of the original mos gene reading frame allows the TGA codon to be bypassed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982041 TI - Adenovirus 3 fiber polypeptide gene: implications for the structure of the fiber protein. AB - The nucleotide sequence of a 1,330-base-pair-long DNA segment located between map coordinates 88.5 and 92.3 in the adenovirus type 3 (Ad3) genome was determined. Transcripts from the r-strand of the region were mapped by S1 nuclease analysis and by in vitro translation of RNA, selected by filter hybridization. The results revealed that the sequenced region encodes the Ad3 fiber polypeptide with a molecular weight of 34,800. A comparison between the predicted amino acid sequences of the Ad3 and the Ad2 fiber polypeptides revealed that they have almost identical secondary structures, consisting of a tail, a shaft, and a knob. A striking difference between Ad2 and Ad3 fibers was that the shaft of the Ad3 fiber was significantly shorter, containing only 6 repeat units compared with 22 in the Ad2 fiber. The secondary structure suggests that the fiber is a dimeric structure, as proposed earlier (N.M. Green, N.G. Wrigley, W.C. Russell, S.R. Martin, and A.D. McLachlan, EMBO J. 2:1357-1365, 1983), with the size of the polypeptide determining the length of the fiber protein. PMID- 2982042 TI - Construction of a viable simian virus 40 variant that carries a poly[d(GT) . d(CA)] insertion. AB - A 90-base-pair tract of a simple sequence composed of alternating guanosine and thymidine nucleotide residues (poly[d(GT) . d(CA)]) was inserted into the simian virus 40 genome at nucleotide 2666 (0.17 map units). The poly[d(GT) . d(CA)] insertion was stably maintained in the viral genome, but the variant virus grew more slowly than simian virus 40. PMID- 2982043 TI - Histological patterns of treatment failures in testicular neoplasms. AB - We reviewed 77 consecutive autopsies performed between 1965 and 1982 on patients who had been treated for germ cell tumors of the testis at our institute. Identifiable germ cell tumor was present at autopsy in 64 cases. On review, a single pattern was seen at autopsy in the majority of the cases (69.7 per cent) compared to the primary tumors, in which single patterns were seen in only 45 per cent. The occurrence of yolk sac tumor as the sole element in 6 of 29 autopsy specimens of nonseminomatous tumors after the introduction of the current standard 3-drug therapy and only once in 32 autopsies before 1976 appears significant. A possible explanation for this finding is that the yolk sac element was obscured by more aggressive and rapidly growing varieties of tumor in the earlier years but proved less responsive to chemotherapy. PMID- 2982045 TI - Fulminant varicella hepatitis following bone marrow transplantation. PMID- 2982044 TI - Rapidly expanding right renal mass in a 12-year-old boy. PMID- 2982046 TI - Leads from the MMWR. Prevalence of cytomegalovirus excretion from children in five day-care centers. PMID- 2982047 TI - Preventing neonatal herpes. PMID- 2982048 TI - Fulminant hepatic failure caused by genital herpes in a healthy person. PMID- 2982049 TI - Significant role of alpha 2-adrenoceptors in coronary artery spasm. AB - Ergonovine, methoxamine and clonidine were administered intravenously in order to clarify the mechanism of coronary artery spasm in canine experiments. After these administrations, 27.5% of the dogs treated with ergonovine, 59.5% of the dogs treated with methoxamine and 72.7% of the dogs treated with clonidine showed ST segment elevation. The combined application of propranolol with ergonovine or methoxamine could more easily induce ST segment elevation than a single application of ergonovine or methoxamine. Most cases of elevated ST segment returned to the normal after the administration of nitroglycerin and phentolamine. Yohimbine was completely effective for the restoration of normal ST segment. Plasma norepinephrine decreased to 20 +/- 8.0% of the control when ST segment elevation was induced. This is a significantly greater decrease than that seen without ST segment elevation (p less than 0.001). Concomitant with the restoration of normal ST segment, plasma norepinephrine increased after the administration of yohimbine. Methoxamine and clonidine increased (p less than 0.01) and yohimbine decreased (p less than 0.01) coronary vascular resistance. Coronary spasm was visualized on coronary arteriogram after the application of clonidine; also coronary vasodilation was visualized after the administration of yohimbine. These results confirm the hypothesis that coronary artery spasm is mediated by pre- and postsynaptic alpha 2-adrenoceptors. PMID- 2982050 TI - Plasma thromboxane B2, 6-keto PGF1 alpha and cyclic nucleotides levels as related to treadmill exercise test in patients with ischemic heart disease. AB - Plasma prostanoids and levels of cyclic nucleotides were studied in forty-nine outpatients with precordial pains and subjected to the treadmill exercise test. Blood samples were drawn before, immediately after and 30 minutes after exercise, from antecubital veins. Plasma TXB2, 6-keto PGF1 alpha, cyclic AMP and cyclic GMP levels were measured by radioimmunoassay. The results of exercise tests were evaluated according to the treadmill exercise score (TES) of Hollenberg et al. Patients were divided into two groups; those with TES (-) with ischemic findings in exercise and those with TES (+) with normal exercise response. There were no differences in TXB2 levels between the two groups before exercise. Immediately after exercise statistically significant differences in TXB2 levels were present between the two groups. There were increased levels in the TES (-) group and decreased levels in the TES (+) group (p less than 0.01). Although the 6-keto PGF1 alpha levels were the same between the two groups before exercise, 6-keto PGF1 alpha levels in the TES (+) group increased significantly immediately after exercise. Similar changes were noted in the case of cyclic nucleotides, and the differences increased immediately after exercise. We conclude that high responses of cyclic nucleotides and PGI2 are required to counteract increases in levels of TXA2 and diminution of these responses may be an important phenomenon involved in the physiological status of ischemic heart disease. PMID- 2982051 TI - Serum concentration and effects of a single dose of enalapril maleate in patients with essential hypertension. AB - The antihypertensive effect of a non-sulfhydryl, long acting ACE (angiotensin converting enzyme) inhibitor, MK-421, was evaluated by administering a single dose of 10 mg to 13 patients with mild to moderate essential hypertension. The pharmacokinetic profile of MK-421 and its potent active metabolite, MK-422, was also assessed, together with the effect on the various components of the renin angiotensin system. A single dose of MK-421 produced a significant fall in MBP from 2 to 24 hours post-drug. As could be expected, plasma ACE activity was suppressed up to 24 hours after MK-421. The half-life of MK-422, Cmax and [AUC]24(0) of MK-421 and MK-422 were measured. No significant change in plasma bradykinin or urinary excretion rate of kallikrein was observed, whereas a slight increase was observed in the urinary excretion rate of kinins after MK-421 in 8 patients. Significant correlations were observed between pretreatment PRA levels and the maximum fall in MBP. PMID- 2982052 TI - Immunologic studies with mouse mammary tumor viruses: comparative studies with spontaneous mammary tumor cell vaccines from five inbred mouse strains. AB - Vaccines were prepared from cells of primary spontaneous mammary tumors (MT) of RIII/Imr, GR/Imr, C3H/Imr, A/Imr, and C3HfC57BL/Imr mice. Each vaccine was used to immunize "murine mammary tumor virus (MuMTV)-free" BALB/c/Imr and C57BL/Imr mice before challenge with infectious RIII-MuMTV, GR-MuMTV, C3H-MuMTV, and A MuMTV. RIII-MT and C3H-MT cells protected mice against tumorigenesis by all four MuMTV's; GR-MT cells protected against all but the RIII-MuMTV challenge; A-MT cells were ineffective against challenge by all four MuMTV's and significantly enhanced development of A-MuMTV-induced tumors. The activity of the C3H-MT cells was not seen when C3HfC57BL cells, free of the standard C3H-MuMTV, were used. The same cell vaccines were inoculated into the five donor strains of mice in attempts to prevent spontaneous MT in these high-incidence strains, which are naturally infected with MuMTV at birth. None of the vaccines prevented tumorigenesis in these mouse strains although delays in the appearance of tumors were observed in RIII and GR mice inoculated with either isologous MT cells or C3Hf-MT cells. The role of viral antigens acting alone or in concert with cellular antigens is discussed. PMID- 2982053 TI - Collagenase secretion by human breast neoplasms: a clinicopathologic investigation. AB - Evidence is presented that biopsy specimens from fibroadenomas, benign cystic lesions, and carcinomas of the human breast can produce in organ culture a neutral protease capable of digesting type I collagen. This enzyme activity, measured with the use of a radioactive release assay, was characterized as true vertebrate collagenase and occurred in both active and latent (requiring trypsin activation) forms. For the two types of benign breast lesion studied, collagenase secretion was significantly higher from fibroadenomas than from benign cystic tissue. Breast carcinomas, however, exhibited a wide quantitative spectrum of collagenase secretion, encompassing the extremes observed for the benign lesions and showing no correlation with histologic type. These results, while providing a plausible mechanism for the marked collagen degradation seen in disseminating neoplasms, demonstrate that high collagenase secretory activity is not pathognomonic of invasive behavior. The findings, however, indicate disordered regulation of collagenase activity in malignant tumors. PMID- 2982054 TI - Isolation of transforming and early antigen-inducing Epstein-Barr virus from nasopharyngeal carcinoma hybrid cells (NPC-KT). AB - The biologic activities of Epstein-Barr (EB) virus from an epithelioid nasopharyngeal carcinoma hybrid cell line (NPC-KT) and three subclones from the NPC-KT cells were examined. Much infectious virus was released by treatment with 5-iodo-2'-deoxyuridine. All virus preparations from NPC-KT cells and the subclones were found to possess both transforming and EB virus-induced early antigen (EA)-inducing activities. The lymphoblastoid cell lines, which were established by infection of human cord-blood lymphocytes with the virus, were diploid with normal karyotypes. The cell lines were positive for EB virus associated nuclear antigen but negative for EA and EB viral capsid antigen. PMID- 2982055 TI - Comparison between murine natural antibodies and natural killer cells: recognition of separate target structures as revealed by differential in vitro expression and dependence on glycosylation. AB - The specificity of complement-fixing, cytotoxic antibodies against the YAC lymphoma in sera of normal young adult (A X C57BL)F1 mice was studied. In vivo maintained, immunoselected sublines of the YAC lymphoma expressed low amounts of the natural antibody (NAb) target structure. These cell lines were also resistant to natural killer (NK) cell-mediated lysis. After 2-3 weeks of in vitro culture the immunoselected cell lines became NK sensitive, but they remained resistant to NAb. When several independently derived variants selected for low NK sensitivity were tested for their ability to absorb NAb, the degree of absorption varied considerably among the variants. NAb could be inhibited by purified C-type virus particles and also by bacterial sonicates and various glycoprotein preparations. Treatment of target cells with tunicamycin, an inhibitor of asparagine-linked glycosylation, decreased the sensitivity to NAb lysis but had no impact on NK sensitivity. Thus the results indicated that a) NAb and NK cells recognized separate target structures and b) the target structure(s) for NAb but not for NK cells were saccharides of the N-glycosidic type. PMID- 2982056 TI - Proteolytic enzymes in tumor metastasis. II. Collagenase type IV activity in subcellular fractions of cloned tumor cell populations. AB - Collagenase type IV degradation activity was examined in metastatic and nonmetastatic clones of the Lewis lung carcinoma (3LL) and of the T10 sarcoma. Conditioned media prepared from cells of both tumors grown in vitro contained low degradation activities, whereas conditioned media from organ cultures of the same clones grown as solid tumors in animals exhibited higher degradation activities. Analysis of subcellular fractions of tumor cells showed that collagenase type IV activity was localized mainly in the cytoplasmic fraction. Crude homogenates or detergent lysates manifested low degradation activities. Little activity was associated with purified plasma membrane preparations and endoplasmic reticular fractions. However, addition of plasma membrane to conditioned media and to cytoplasmic fractions reduced the degradation activities of the cytoplasmic fractions. Possibly a factor that inhibits collagenase type IV exists in the cells in a vesicular form. No correlation between degradation activity and metastatic capacity was demonstrated in the models used in this study. Both metastatic and nonmetastatic clones of the same tumor similarly could degrade basement membrane components. PMID- 2982057 TI - Effects of ethanol on the chemical and structural properties of biologic membranes. PMID- 2982058 TI - Estrogen and thyroid-stimulating hormone (TSH) receptors in neoplastic and nonneoplastic human thyroid tissue. AB - Estrogen-binding receptors (ER) and thyroid-stimulating hormone (TSH) receptors were observed in the cytosol and in a membrane particulate fraction, respectively, in most neoplastic and nonneoplastic human thyroid tissues. Fourteen of 15 thyroid neoplasms and 6 of 15 nonneoplastic thyroid specimens had estrogen receptors (assuming the sensitivity of our estrogen receptor assay is 0.2 fmole/mg protein), and 14 of 15 thyroid neoplasms and 11 of 15 nonneoplastic thyroid specimens had a high affinity, low capacity TSH receptor. Neoplastic thyroid tissue had more ER (2.35 +/- 0.70/fmole/mg protein) than nonneoplastic thyroid tissue (0.57 +/- 0.181/fmole/mg protein) removed from the same patients (P less than 0.05). The Kd for ER did not differ in nonneoplastic (0.41 +/- 0.090 nM) and neoplastic (0.311 +/- 0.048 nM) thyroid tissue. The number of TSH receptors was comparable in neoplastic (0.609 +/- 0.191 pmole/mg protein) and in nonneoplastic (0.765 +/- 0.181 pmole/mg protein) thyroid tissue removed from the same patients who had the ER studies. The maximal adenylate cyclase response to TSH was greater in the neoplastic (147 +/- 26.9 pmole/mg protein/30 min) than in nonneoplastic thyroid tissue (32.8 +/- 6.69 pmole/mg protein/30 min) (P less than 0.001) suggesting a greater metabolic responsiveness of the neoplastic thyroid tissue to TSH. No correlation was evident, however, between the number of estrogen and TSH receptors in nonneoplastic and neoplastic thyroid tissue (r = 0.226). This study demonstrates that neoplastic human thyroid tissues have both estrogen receptors and TSH receptors. The neoplastic tissue also has a greater AC response to TSH than nonneoplastic thyroid tissue. PMID- 2982059 TI - Myoepithelioma of the head and neck: case report and review. AB - The clinical, light, and ultrastructure features of a myoepithelioma occurring on the hard palate of a 24-year-old woman are presented and compared with 41 myoepithelioma of the head and neck described in the English literature. These 42 tumors (39 benign and 3 malignant) occurred in individuals from 14 to 81 years of age, affected both sexes about equally, and most often presented as a slowly enlarging, asymptomatic mass. The parotid gland and palate were the most common sites of occurrence. The tumors are typically circumscribed and encapsulated, vary from 1 to 5 cm in the greatest dimension, and are composed of spindled and/or plasmacytoid myoepithelial cells. Prognosis correlates with histologic appearance and parallels that of the pleomorphic adenoma. Conservative excision with a margin of uninvolved tissue is curative. The differential diagnosis and histogenic relationship with other closely related neoplasms are discussed. PMID- 2982060 TI - Primary mucinous adenocarcinoma of the trachea: the case for complete surgical resection. AB - Adenocarcinomas of the trachea are rare but important because the only reported treatment in the past (radiation) has resulted in uniformly poor results. The authors recently had the opportunity to treat a young woman with a mucin producing adenocarcinoma of the trachea. A sleeve resection of the involved trachea with reanastomosis was successful, and the patient is alive and well with no evidence of tumor four years later. Thus, primary resection should be strongly considered for adenocarcinoma of the trachea whenever possible. PMID- 2982061 TI - Immunological status of trophoblastic neoplasia. AB - Six cases of choriocarcinoma, 17 of vesicular mole, 20 normal pregnant females in the first trimester of pregnancy, and 20 normal nonpregnant women wih previous normal obstetrical history were evaluated quantitatively for cell-mediated immunity by response to 2:4 dinitrocholorobenzene and recall antigens. Serum immunoglobulins were estimated in all cases. Suppression of cell-mediated immunity was marked in choriocarcinoma as compared to vesicular mole. Serum IgG levels were significantly diminished in cases of vesicular mole, indicating diminished humoral immunity. Follow-up after 3 months of treatment with surgery and chemotherapy resulted in marked improvement of cell-mediated immunity. PMID- 2982062 TI - An enzymatic method for the consistent production of monodispersed viable cell suspensions from human solid tumors. AB - An enzymatic method is described for disaggregation of viable tumor cells from human solid tumors. The enzymatic cocktail consists of 0.1% collagenase, 0.01% hyaluronidase, and 0.002% deoxyribonuclease. After mechanical mincing of the tumor tissue, tumor specimens are dissociated by incubation in the enzymatic cocktail for 12-18 hours at room temperature. In 17 cases of sarcoma, the mean yield was 5 X 10(6) viable cells per gram tumor tissue. Yield was 1 X 10(7) viable cells per gram tumor tissue in 23 cases of gastrointestinal carcinoma. The viabilities of tumor cell suspensions ranged from 50 to 98%, except for low viabilities in four specimens that were grossly composed almost entirely of necrotic tissue. The dissociation procedure is simple and the viable cell yield is sufficient for applications in studies of human cancer immunobiology. PMID- 2982063 TI - The significance of metastasis to the bones and soft tissues of the hand. AB - We have treated six patients with cancer metastatic to the hand. All patients died within 6 months after the appearance of hand metastasis. We polled the combined membership of the American Association for Surgery of the Hand and the American Society for Surgery of the Hand to determine their experience with this problem. We also reviewed the pertinent literature concerning this subject. The information obtained from the literature and the poll confirmed our experience. We have managed these patients by amputation, feeling that reconstructive procedures were not indicated if the time necessary to rehabilitate the patient after reconstruction exceeds the patient's life expectancy. PMID- 2982064 TI - Breast carcinoma at the extremes of age: a comparison of patients younger than 35 years and older than 75 years. AB - We assessed the prognosis of patients with breast carcinoma at the extremes of the age distribution of the disease. The groups examined were 166 women 35 years or less and 169 at least 75 years old. Analysis of recurrence and survival showed no significant difference between the groups as a whole or when they were stratified by nodal status. Life expectancy of elderly women with breast carcinoma was significantly reduced when compared with a "normal" age-matched population. Medullary carcinoma was more frequent in young women while a relatively higher proportion of colloid and invasive lobular carcinoma occurred in elderly women. Bilateral carcinoma was found with nearly equal frequency in both age groups. However, elderly women were more likely to have been treated previously for contralateral carcinoma while young women tended to develop asynchronous, subsequent carcinoma. PMID- 2982065 TI - Sequential internal mammary artery grafts. Expanded utilization of an ideal conduit. AB - The internal mammary artery, when used as a conduit for coronary artery bypass, offers a better long-term patency rate and survival rate than the saphenous vein; however, its utility has been limited. Among other factors, the availability of only two internal mammary arteries for anastomosis has been a major limitation. In an attempt to overcome this limitation, we constructed sequential internal mammary artery grafts in 87 patients. In 49 patients (Group I), only one internal mammary artery was used for sequential anastomosis. In another 31 patients (Group II), one internal mammary artery was used for sequential anastomosis and the other was used for single end-to-side anastomosis. Both internal mammary arteries were used in seven patients (Group III) for the construction of sequential anastomoses. Postoperatively, 64 patients were evaluated by exercise stress tests. None of these patients had a positive stress test although seven patients (11%) had electrocardiographic changes that were considered equivocal. Coronary angiography was performed in 35 of the 87 patients, with 92 vein grafts and 90 internal mammary artery anastomotic sites evaluated within 1 year of operation. A total of 83 vein grafts and 84 internal mammary artery anastomotic sites evaluated within 1 year of operation. A total of 83 vein grafts and 84 internal mammary artery anastomoses were found to be patent. Thus the patency rate for vein grafts was 90% and for internal mammary artery grafts, 93%. During the follow-up period (8 to 52 months), three patients died and one was lost to follow up. Among the remaining patients, 79 had complete relief from symptoms, three had minimal symptoms, and one patient obtained no relief from symptoms. Based on these results, we have concluded that the extended use of internal mammary artery, constructing sequential anastomoses, is technically feasible and provides adequate perfusion to the area of myocardium supplied by such grafts. PMID- 2982066 TI - Adenoid cystic carcinoma of the head and neck: a clinical study of 37 cases. AB - Thirty-seven cases of adenoid cystic carcinoma treated at the Otolaryngologic Clinic of the University of Florence between 1962 and 1979 were studied. The patients were treated with radical surgical excision (20), non-radical surgery associated with radiotherapy (13), or with palliative treatment (4). The aim of this study is to evaluate the course of the disease according to the site and type of operation. Our results show that non-radical surgical treatment associated with radiotherapy seems to give results comparable to those obtained with radical surgery alone. PMID- 2982067 TI - Modulation of acetylcholine release from rat striatal slices by the GABA/benzodiazepine receptor complex. AB - GABA, THIP and muscimol enhance spontaneous and inhibit electrically induced release of tritium labelled compounds from rat striatal slices which have been pre-labelled with 3H-choline. Baclofen is inactive in this model. Muscimol can inhibit electrically induced release of tritiated material by approximately 75% with half maximal effects at 2 microM. The response to muscimol can be blocked by the GABA antagonists bicuculline methobromide, picrotoxin, anisatin, R 5135 and CPTBO (cyclopentylbicyclophosphate). Drugs which act on the benzodiazepine receptor (BR) require the presence of muscimol to be effective and they modulate the effects of muscimol in a bidirectional manner. Thus BR agonists enhance and inverse BR agonists attenuate the inhibitory effects of muscimol on electrically induced release. Ro15-1788, a BR antagonist, does not modulate the inhibitory effects of muscimol but antagonizes the actions of clonazepam, a BR agonist, and of DMCM, an inverse BR agonist. These results demonstrate that a GABA/benzodiazepine receptor complex can modulate acetylcholine release from rat striatal slices in vitro. PMID- 2982068 TI - The effects of vasopressin and catecholamines on cyclic AMP in homogenates of rat brain. AB - Vasopressin is known to mediate its action on the kidney through increasing the concentrations of cyclic AMP. As vasopressin is widely distributed in many extra hypothalamic areas of the brain and can be shown to act centrally, we have investigated the effect of vasopressin on cyclic AMP levels in homogenates of the striatal and locus coeruleus areas. In contrast with the effect obtained on the kidney, vasopressin did not stimulate adenyl cyclase activity in rat brain homogenates in a dose-related manner. The stimulation of cyclic AMP observed with dopamine or noradrenaline in these brain areas and the hippocampus was not affected by the presence of vasopressin. These observations suggest that the action of vasopressin on the brain is not mediated through cyclic AMP. PMID- 2982069 TI - Sex differences in the response of rats to drugs affecting GABAergic transmission. AB - The administration of diazepam 1.0 mg/kg decreased the level of plasma corticosterone in female but not in male Wistar rats. Picrotoxin, another drug affecting GABAergic transmission, also brought about an increase of plasma corticosterone in both sexes. However, in order to achieve a plasma corticosterone increase of similar magnitude (more than 500%) a threefold higher dose of picrotoxin had to be given to males. When the convulsive properties of picrotoxin were tested, it became evident that the dose of picrotoxin (2.5 mg/kg) which was subconvulsive in male was almost 100% convulsive in female rats. The existing sex differences in the response of rats to drugs affecting GABAergic transmission might have possible implications in the treatment of GABA system dysfunction. PMID- 2982070 TI - Selective inhibition of cardiac cyclic nucleotide phosphodiesterases by doxorubicin and daunorubicin. AB - Doxorubicin and daunorubicin, the anthracycline antitumor agents, were evaluated for their in vitro and in vivo effect on phosphodiesterase (PDE) activity in mouse tissues. Doxorubicin at a concentration of 10(-4)M inhibited cardiac c-AMP (adenosine 3',5', monophosphate) PDE activity 50% of the control whereas in lungs and spleen, the activity was inhibited only 20%. On the contrary no effect was seen in kidney and liver. In addition, cardiac c-GMP (guanosine 3',5' monophosphate) PDE appeared less sensitive to doxorubicin than c-AMP PDE though inhibition in heart was more pronounced than in any other tissue. It appears that daunorubicin inhibits c-AMP PDE activity in heart markedly less than doxorubicin. Kinetic studies indicate that both inhibitions of c-AMP and c-GMP PDE by doxorubicin were non-competitive with substrate. Intravenous administration of 20 and 30 mg/kg of free doxorubicin to CDF1 mice resulted in 33 and 39% decreases in cardiac c-AMP PDE activity respectively by 72 hrs. In contrast, similar intravenous injections of same doses of doxorubicin entrapped in cardiolipin liposomes had no effect on c-AMP PDE activity in any tissues. These studies demonstrate the relative selectivity of the cardiac cyclic nucleotide PDE inhibitory effect of doxorubicin suggesting that this inhibition might be one aspect of the mechanism of anthracycline-induced cardiotoxicity. PMID- 2982071 TI - Analog specificity of the thyrotropin-releasing hormone receptor in the central nervous system: possible clinical implications. AB - TRH has rapid-onset (30 sec), slow-offset (1-12 days) clinical benefit in patients with amyotrophic lateral sclerosis and other motor neuron disorders. This benefit is probably receptor-mediated and may have at least 2 components. To obtain a better understanding of the various responses to TRH of the spinal lower motor neurons (LMNs) in patients, and possibly to help guide selection of additional therapeutic agents, we utilized rat CNS (spinal-cord and brain membranes) to analyze the ability of certain molecules to inhibit specific binding of [3H]methyl TRH [( 3H]MeTRH) to the TRH receptor. We found: a) lack of high-affinity binding of the TRH-analog DN-1417 by spinal-cord and brain TRH receptor, despite its known strong TRH-like action physiologically on LMNs; b) lack of high-affinity binding of the TRH-product cyclo(His-Pro) by spinal-cord and brain TRH receptor despite its having some strong TRH-like physiologic actions on the CNS; and c) lack of any identifiable high-affinity receptor for cyclo(His-Pro) in spinal cord and brain. From these data we hypothesize that the acute transmitter-like action of DN-1417, TRH, and possibly other TRH-analogs and products on LMNs is via a non-TRH receptor, such as an amine or amino acid neurotransmitter receptor, e.g. a 5-hydroxytryptamine receptor. We further postulate that the CNS TRH-receptor may modulate a trophic-like influence of TRH on LMNs. PMID- 2982072 TI - A cardiac specific analog of angiotensin I potentiated by converting enzyme inhibition. AB - [1-Sarcosine, 7-Alanine] angiotensin I [( 1-Sar, 7-Ala] AI) and closely related analogs were tested for inotropic activity in the isolated cat heart, and for pressor activity in the intact conscious sheep both before and during converting enzyme inhibition (CEI). [1-Sar, 7-Ala] AI exhibited potent inotropic activity but was only weakly pressor. [1-Sar] AI, [1-Sar, 5-Val] AI, [1-Sar, 7-alpha MeAla] AI [1-Sar, 5-Val, 7-NMeAla] AI and [1-Sar, 5-Val, 7-Sar] were all potent agonists in both preparations. The action of [1-Sar, 7-Ala] AI was potentiated by CEI in both the isolated heart and the intact sheep. The activity of the remaining analogs was either partially or completely blocked by CEI. The activity of all analogs was inhibited by AII receptor blockade. These data indicate that the nature of the substitution in position 7 determines the affinity of the analog for converting enzyme. The [7-Ala] substitution appears to decrease the effect of the analog upon vascular receptors. PMID- 2982073 TI - Rapid and convenient separation of phospholipids and non phosphorus lipids from rat heart using silica cartridges. AB - The separation of non phosphorus lipids and phospholipids of rat heart using Sep pack Silica cartridges is described. No cartridge preparation is necessary before utilization. The separation of lipid extracts is very fast. A complete partition of non phosphorus lipids and phospholipids is obtained. PMID- 2982075 TI - [Studies with 99mTc-pyrophosphate of reparative osteogenesis during treatment of closed oblique fractures of the long tubular bones]. AB - Altogether 56 patients aged 20-55 with closed oblique fractures of the upper and lower limb bones were observed. Reparative osteogenesis was studied using 99mTc pyrophosphate. An elevated RP accumulation at the ends of the bone splinters was found from the 1st day after fracture. By the end of the 1st month after fracture the RP activity increased up to 750% in the absence of splinter dislocation. In dislocation it was 50-80% less. The RP maximal activity in the place of fracture was noted on the 7-8th week, in the dislocation of splinters on the 9-10th week. By the results of radionuclide studies, the fracture consolidated right after a decrease in the 99mTc-pyrophosphate accumulation that made it possible to determine the optimum time of bone consolidation. PMID- 2982074 TI - [Radionuclide diagnosis of ovarian tumors]. AB - The authors present a method with the use of 99mTc-pyrophosphate for differential diagnosis of malignant and benign ovarian tumors. A total of 46 patients were examined. Of these, the sample hyperfixation in the zone of malignant ovarian tumors with the rate of differential accumulation of 200-260% was observed in 7. In benign tumors and inflammatory ovarian changes, radiopharmaceutical hyperfixation was either not observed or it did not exceed 130%. Diagnosis in all the patients was verified at operation with subsequent histological examination. PMID- 2982076 TI - [Electro-roentgeno-thyrolympho-scanography in the diagnosis of thyroid diseases]. AB - The results of examination of 79 patients of whom 25 were thyroid cancer patients, showed that electroroentgenothyrolymphoscans made it possible to judge with a sufficient degree of significance of the size, site and function of the intrathyroid lymph nodes of which some were "hidden" because of their small size on routine scans. This method can be successfully used for differential diagnosis of different thyroid diseases including tumors alongside with other morphological methods. PMID- 2982077 TI - [Radionuclide studies of the functional state of the bone marrow]. PMID- 2982078 TI - [Radionuclide studies on healing in closed transverse fractures of the long tubular bones]. AB - A total of 86 patients were examined by 99mTc-pyrophosphate in the course of therapy using the transosseous synthesis method. An elevated accumulation of the radiopharmaceutical was observed since the first day following a trauma. Maximum enhancement of metabolic processes in the absence of fragment displacement was noted on the 8th week and in the lack of displacement on the 9th-10th week. Then the intensity of bone formation got weaker. It was found appropriate that G. A. Ilizarov's apparatus should be removed exactly at that time. The content of mineral components in the regenerate was over 60% which demonstrated the bone ability to stand physiological loads. PMID- 2982079 TI - Phosphotyrosine and phosphoprotein phosphatase activity of alkaline phosphatase in mineralizing cartilage. AB - We used embryonic skeletal cartilage known to have high levels of alkaline phosphatase activity to determine whether growing cartilage has phosphotyrosine phosphatase activity and phosphotyrosinyl histone phosphatase activity at physiologic pH. Embryonic chick pelvic cartilage and fetal pig scapular growth plate cartilage were assayed using phosphotyrosine as substrate at pH 7.5 and the amount of tyrosine generated measured. Both cartilage models had Km for phosphotyrosine between 6 to 24 mus mol/L. Phosphotyrosine phosphatase activity correlated with alkaline phosphatase activity as assessed by (1) distribution of histologic staining for alkaline phosphatase within the cartilages, (2) hormonal stimulation of cartilage alkaline phosphatase activity in vitro, (3) comparison of alkaline phosphatase and phosphotyrosine phosphatase activities in the presence of known inhibitors (vanadate, levamisole, homoarginine, and zinc), and (4) assaying chick epiphyseal cartilage alkaline phosphatase purified to homogeneity for phosphotyrosine phosphatase activity. Areas of cartilage with elevated alkaline phosphatase activity also had raised phosphotyrosine phosphatase activity. Triiodothyronine, a known stimulator of cartilage alkaline phosphatase, increased chick cartilage alkaline phosphatase activity 88% and phosphotyrosine phosphatase activity 106%, and stimulated porcine growth-plate cartilage alkaline phosphatase activity 91% and phosphotyrosine phosphatase activity 145% after 3 days of in vitro incubation. Each of the inhibitors block alkaline phosphatase and phosphotyrosine phosphatase activities. The purified alkaline phosphatase had a Km for phosphotyrosine of 18 mus mol/L and Vmax of 5700 nmol tyrosine/mg protein/h, which is well over 1000-fold higher than the phosphotyrosine phosphatase activity found in the above preparations of pelvic and scapular cartilage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982080 TI - Cytomegalovirus: the challenge of a common virus infection. PMID- 2982081 TI - Prevalence of cytomegalovirus antibodies in obstetric nurses. A study in a specialist metropolitan teaching hospital. AB - The prevalence of antibodies to cytomegaloviruses (CMV) in applicants for nursing staff appointments was shown to be significantly lower than that reported earlier in obstetric patients of the same metropolitan area. Age and previous nursing experience with midwifery patients or neonates were found to be significant factors associated with the prevalence of antibodies to CMV in staff applicants. Student midwives younger than 25 years were the group most susceptible to primary CMV infection. The duties that seronegative pregnant staff members or those planning pregnancy at an early date should be allowed to carry out are discussed. The importance of hygiene in the prevention of nosocomial infections, and of the skin care of hands subjected to frequent washing, is stressed. PMID- 2982082 TI - Antibodies to adult T-cell leukemia virus (ATLV/HTLV-I) in AIDS patients and people at risk of AIDS in Germany. AB - A total of 2048 serum samples from Germany were examined for antibodies to adult T-cell leukemia virus (ATLV) structural polypeptides with an enzyme-linked immuno sorbent assay (ELISA) and confirmative immuno precipitation. The origin of the sera samples was: 850 samples taken for virological or protozoal diagnosis; 626 samples from male homosexuals, about 20% of whom had lymphadenopathy syndrome; 164 from hemophiliacs; 184 were from multiple transfused, mostly dialysis patients; 9 from intravenous drug abusers; 182 from suspected cases of acquired immuno deficiency syndrome (AIDS) and 33 from AIDS-patients. In none of these sera did we detect antibodies to ATLV, except in the serum of one patient who had been on hemodialysis for over 11 years. Obviously infection with ATLV or a serologically related agent is very rare in our country and an association with AIDS could not be observed. PMID- 2982083 TI - Transduction of virulence in herpes simplex virus type 1 from a pathogenic to an apathogenic strain by a cloned viral DNA fragment. AB - Herpes simplex virus (HSV) type 1 strain HFEM, the genome of which contains a deletion of about 4 kilobasepairs (kbp) between 0.7 and 0.8 viral map units, is apathogenic in the tree shrew when the animals are inoculated intravenously, intraperitoneally and/or subcutaneously. Similar results were obtained using Balb/c mice. Studies of the state of viral latency in animals infected with HSV-1 strain HFEM revealed that this strain was unable to colonize the ganglia of tree shrews. Infectious virus could be recovered only from the spleens of latently infected tree shrews. Thus, this system offers new opportunities for investigating the gene functions responsible for the virulence of HSV-1. Marker rescue experiments were performed by the contransfection technique using native DNA of HSV-1 strain HFEM and Bam H I DNA fragment B derived from the pathogenic HSV-1 strain F and cloned in a bacterial vector. A number of different intratypic recombinants were established in which the deleted region of HSV-1 strain HFEM had been repaired. The pathogenicity of these recombinants was examined in vivo. One of the recombinants (HSV-R-HFehx-C19) caused generalized and lethal herpes virus infection in juvenile and adult tupaias, indicating that the virulence of the pathogenic HSV-1 strain F can be transduced by the cloned Bam H I DNA fragment B to the apathogenic HSV-1 strain HFEM. PMID- 2982084 TI - Role of radionuclide imaging in the diagnosis of chondrosarcoma. AB - The diagnosis of chondrosarcoma may be difficult if there is an atypical radiographic appearance or an inconclusive biopsy. Radionuclide bone scans of 13 patients with chondrosarcoma were reviewed to assess if a pattern of scan features could be recognised in association with this tumour. A combination, including increased blood pool activity, moderate intensity of uptake, patchy uptake with cortical predominance of activity, minimal distortion of bony outline, and a well-defined scintigraphic margin, occurred regularly in the series. Recognition of this characteristic pattern of scintigraphic features in cases of suspected chondrosarcoma may assist in the diagnostic assessment. PMID- 2982085 TI - Partial nephrectomy for Wilms' tumor in a child with hemihypertrophy. PMID- 2982086 TI - Diagnostic and therapeutic applications of a betamimetic agent in infertility. PMID- 2982087 TI - Preventing lead poisoning in young children--United States. PMID- 2982088 TI - Age-related alterations in the catecholamine-sensitive adenylate cyclase system of the prostate. AB - Considerably reduced responses to stimulation by isoproterenol of adenylate cyclase activity of prostatic membranes were observed in 12- to 18-month-old rats, compared to 3-month-old animals. Plasma testosterone levels were significantly lower in 18-month-old rats, while 12-month-old animals showed levels similar to those present in young ones. A decrease in isoproterenol activation of adenylate cyclase was not associated with a fall in beta-adrenergic receptor sites. Guanine triphosphate and 5'-guanylylimidodiphosphate (Gpp(NH)p) were effective in potentiation of isoproterenol activation of adenylate cyclase and altering the affinity of beta-adrenergic receptors for the agonist in membranes from young rats but not from the aged. The age-induced refractoriness to isoproterenol or Gpp(NH)p was observed without a significant loss in NaF stimulated activity. Prior incubation of aged membranes with isoproterenol and GMP restored subsequent stimulation by Gpp(NH)p, presumably due to the clearance of inhibitory GDP tightly bound to the guanine nucleotide regulatory components in aged membranes. These results indicate that the dysfunction in the adenylate cyclase system of old prostates may not be related to a modification in the beta adrenergic receptor per se, but to, in part, a defect in the interaction of activating guanine nucleotides with regulatory components of the adenylate cyclase system. PMID- 2982090 TI - Characterization of guinea pig myocardial leukotriene C4 binding sites. Regulation by cations and sulfhydryl-directed reagents. AB - Using [3H]leukotriene C4 (LTC4) and radioligand-binding techniques, specific leukotriene C4 binding sites have been identified in membranes derived from guinea pig ventricular myocardium. High performance liquid chromatography analyses indicated that, in the presence of the gamma-glutamyl transpeptidase inhibitor L-serine-borate (80 mM), less than 2% of membrane-bound [3H]LTC4 was converted at 20 degrees to [3H]leukotriene D4 or [3H]leukotriene E4. The specific binding of 4 nM [3H]LTC4, in the presence of 80 mM L-serine-borate, reached a stable steady state within 15 min at 20 degrees (pH 7.5). A monophasic Scatchard plot of saturation binding data yielded a dissociation constant (Kd) of 27.5 +/- 6.0 nM and a maximum number of binding sites (Bmax) of 19.9 +/- 5.2 pmol/mg of membrane protein. Competition binding studies of [3H]LTC4 with synthetic leukotriene C4, leukotriene D4, and leukotriene E4 and the putative peptidoleukotriene antagonists FPL 55712, SKF 88046, and 4R-hydroxy-5S-1 cysteinylglycine-6Z-nonadecanoic acid revealed an order of potency of leukotriene C4 much greater than 4R-hydroxy-5S-1-cysteinylglycine-6Z-nonadecanoic acid greater than SKF 88046 greater than LTE4 greater than LTD4 greater than FPL 55712. The specific [3H]LTC4 binding was stimulated by the divalent cations Ca2+, Mg2+, and Mn2+ and to a lesser degree by the monovalent cations Na+, K+, Li+, and NH4+. CaCl2 (3 mM) and NaCl (150 mM) stimulated the LTC4 binding by increasing the Bmax to 42.6 +/- 5.9 and 35.0 +/- 2.0 pmol/mg, respectively, but had minimal effects on Kd. Pretreatment of the heart membranes with the sulfhydryl reagent N ethylmaleimide decreased the specific [3H]LTC4 binding in a concentration dependent manner. The N-ethylmaleimide-induced inactivation of [3H]LTC4 binding sites was protected by occupation of the binding site with the agonist leukotriene C4, but no protection was observed with the antagonist SKF 88046. Scatchard analyses of saturation isotherms indicated that 30 microM N ethylmaleimide pretreatment reduced the Bmax of the [3H]LTC4 binding to 8.2 +/- 3.1 pmol/mg with minimal effects on Kd. The data provide direct biochemical evidence for specific [3H]LTC4 binding sites in the guinea pig heart membranes. The [3H]LTC4 binding sites appear to be modulated by divalent and monovalent cations and free sulfhydryl group(s) may be associated with the agonist-binding site. The results suggest that the physiological effects of the leukotrienes on the guinea pig heart may be mediated through membrane-bound receptors. PMID- 2982089 TI - Muscarinic responses and binding in a murine neuroblastoma clone (N1E-115). Mediation of separate responses by high affinity and low affinity agonist receptor conformations. AB - Murine neuroblastoma cells (clone N1E-115) possess muscarinic receptors that mediate multiple responses, including the elevation of cyclic GMP levels and the inhibition of receptor-mediated increases in cyclic AMP. Evidence is presented showing that two muscarinic agonist-receptor conformations in N1E-115 cells each separately mediate a cyclic nucleotide response. Pirenzepine inhibited the [3H]cyclic GMP response to carbachol with a KD value of approximately 6 nM, whereas it inhibited the ability of carbachol to reduce prostaglandin E1-mediated elevations in [3H]cyclic AMP levels with a KD value of 93 nM, thus differentiating between two classes of receptors involved in these responses. Ten muscarinic agonists were studied for their ability to mediate the two cyclic nucleotide responses. Six were as effective as acetylcholine in the reduction of [3H]cyclic AMP levels, but only two were as effective as acetylcholine in elevating [3H]cyclic GMP levels. Four agonists (arecoline, pilocarpine, oxotremorine, and McN-A343) were ineffective in increasing [3H]cyclic GMP levels. These four agonists and bethanecol, which could increase [3H]cyclic GMP levels only 18% as well as acetylcholine, behaved as competitive antagonists in this response to carbachol. These partial agonists, in contrast to carbachol, bound to only one class of muscarinic sites in N1E-115 cells with equilibrium dissociation constants determined by competition binding assays which agreed well with their respective EC50 values for their effect on [3H]cyclic AMP levels. The equilibrium dissociation constants for the partial agonists determined by their inhibition of carbachol in the [3H] cyclic GMP response also agreed well with their respective EC50 values for mediating the [3H]cyclic AMP response. Thus, the partial agonists bound to the same receptors at which carbachol mediated [3H]cyclic GMP formation, but with KD values about the same as their respective EC50 values for inhibition of prostaglandin E1-mediated [3H]cyclic AMP increases. The full agonists acetylcholine and methacholine, like carbachol, bound to two sites in N1E-115 cells. For the six agonists able to stimulate both responses at least to some degree, the ratio of their potencies at each response correlated with their respective efficacies at each response but with much more dependence in the [3H]cyclic GMP response.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982091 TI - Characterization of the hydrophobic properties of the receptor for 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - The hydrophobic character of the receptor for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been estimated by its tendency to adsorb to hydrophobically interacting matrices. The receptor is adsorbed to uncharged pentyl-Sepharose but not to butyl-Sepharose at 1 M NaCl. It is also adsorbed to phenyl-Sepharose or Cibacron blue-Sepharose at lower ionic strengths (0-0.15 M NaCl). Elution of adsorbed receptor could not be achieved under mild conditions (decreasing salt concentration, increasing glycerol concentration). A concentration of 0.2% (w/v) of the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was required to desorb the receptor from pentyl-Sepharose with an approximate yield of 11-14% of the specific [3H]TCDD-binding activity. The CHAPS treated receptor exhibited the same physicochemical characteristics as that in crude cytosol (4-5 S, Stokes radius approximately 60 A). Furthermore, the effects of detergents other than CHAPS on hydrodynamic parameters and on [3H]TCDD binding to the receptor were studied. In conclusion the TCDD receptor showed more pronounced hydrophobic properties than those reported for steroid hormone receptors. PMID- 2982092 TI - A system to determine basepair substitutions at the molecular level, based on restriction enzyme analysis; influence of the muc genes of pKM101 on the specificity of mutation induction in E. coli. AB - A system has been developed for the analysis of basepair substitutions that are involved in the reversion of a specific missense mutation. The method is based on the ability of restriction enzymes to recognize and cut specific DNA sequences. Wild-type revertants arising from AT----GC transitions, pseudo wild-type revertants arising from AT-transversions and second site revertants can be distinguished. 4 mutagenic agents have been used, 2,6-diaminopurine, MMS, EMS and ENU, which differ in the types of damage they cause in DNA and in the susceptibility of the damage to repair. All 4 mutagens effectively enhanced the reversion of the mutation studied, trpA223, particularly by increasing the fraction of AT----GC transitions. In this system the influence of the muc genes of plasmid pKM101 was investigated. The presence of these genes reduced the fraction of AT----GC transitions and enhanced the fraction of AT-transversions as well as the fraction of second-site mutations. This change in mutation specificity is found irrespective whether mutation induction occurs mainly via SOS repair (MMS, ENU) or via mainly misreplication (2,6-diAP, EMS). These data suggest that the muc genes are involved in the induction of mutations not only during SOS repair, but also during misreplication. The change in mutation specificity may be caused by a change in the selection and insertion of nucleotides by the DNA-polymerising complex, or by interference with the repair of mismatched bases. PMID- 2982093 TI - Safety of the hepatitis B vaccine. PMID- 2982094 TI - Superoxide anion generation by alveolar macrophages in Crohn's disease. PMID- 2982096 TI - Pathogenesis of viral infections. Basic concepts derived from the reovirus model. PMID- 2982095 TI - Coincidental appearance of LAV/HTLV-III antibodies in hemophiliacs and the onset of the AIDS epidemic. PMID- 2982097 TI - The AIDS epidemic. PMID- 2982098 TI - Adenovirus infections in patients undergoing bone-marrow transplantation. AB - Viral infection is commonly observed after bone-marrow transplantation. We isolated adenovirus from 51 of 1051 patients undergoing marrow transplantation between 1976 and 1982. Of the 46 isolates available for typing, 13 (27.7 per cent) were of the closely related species 11, 34, or 35 (subgenus B). All 13 of the patients with these species had positive urine cultures. The species have previously been associated with the acquired immunodeficiency syndrome or with renal transplantation but are not commonly found in community surveys. Invasive infection was confirmed by biopsy or autopsy in 10 of 51 patients. Seven of the 10 had virus isolated from lung, and 4 died from pneumonia attributed to adenovirus. Two of the five patients with renal isolates had evidence of virally induced renal impairment, and both patients with liver isolates had adenovirus hepatitis. There was no common source that accounted for these adenovirus infections, and the most likely source of infection appeared to be endogenous viral reactivation. The only identifiable risk factor for the development of infection and for severe disease was the presence of moderate to severe graft versus host disease. PMID- 2982099 TI - Regulation of glutamate receptor binding by the cytoskeletal protein fodrin. AB - The erythrocyte cytoskeleton, which consists primarily of a meshwork of spectrin and actin, controls cell shape and the disposition of proteins within the membrane. Proteins similar to spectrin have recently been found in diverse cells and tissues, and it is possible that they mediate the capping of cell-surface receptors, although this has not been demonstrated directly. In neurones, the spectrin-like protein fodrin lines the cortical cytoplasm and may link actin filaments to the membrane. Fodrin has been hypothesized to regulate the number of receptor binding sites on neuronal membranes for the putative neurotransmitter L glutamate. Micromolar calcium concentrations activate the thiol protease calpain I, induce fodrin degradation and more than double the density of glutamate binding sites; these effects are all blocked by thiol protease inhibitors. We have now used specific antibodies to examine further the role of fodrin proteolysis in regulating glutamate receptors. We report that fodrin antibodies block the fodrin degradation and increase in glutamate binding normally induced by calcium, and so provide direct evidence for control of membrane receptors by a non-erythroid spectrin. PMID- 2982100 TI - Growth of human breast cancer cells inhibited by a luteinizing hormone-releasing hormone agonist. AB - About one-third of human breast cancers require hormones for their continued growth and endocrine ablation or anti-hormone therapy can cause regression of these tumours. As a consequence, ovariectomy in premenopausal women or administration of an anti-oestrogen (tamoxifen) in postmenopausal women represent major options for treatment of metastatic breast cancer. Alternatively, chronic administration of agonistic analogues of luteinizing hormone-releasing hormone (LHRH) causes regression of mammary tumours in experimental animals, and such treatment has shown promise in a small series of premenopausal women with advanced breast cancer. It has been assumed that these results were achieved by suppressing the pituitary-ovarian axis, as the treatment causes a reduction in circulating levels of gonadal steroids similar to that produced by castration. However, LHRH agonists can exert major effects on tissues other than the pituitary in animals and in the human. Such findings, coupled with reports of LHRH in human breast milk and immunohistochemical evidence for the presence of LHRH-like activity in some human breast tumours, prompted us to test whether LHRH agonists could have direct antitumour effects. We now report major direct effects of LHRH and its agonists on the growth of breast tumour cells in culture. PMID- 2982102 TI - Cyclic GMP and neurone death. PMID- 2982101 TI - A retrovirus-like strategy for expression of a fusion protein encoded by yeast transposon Ty1. AB - Eukaryotic transposons such as the Ty element of yeast or the copia-like sequences of Drosophila show structural and functional similarities to both prokaryotic transposons and retroviral proviruses, but the prokaryotic transposons and retroviral proviruses use markedly different expression strategies which yield products having entirely different functions. To determine the phylogenetic relationship between eukaryotic transposons, prokaryotic transposons and retroviruses, we have sought to identify and characterize the proteins encoded by the yeast Ty element and to describe the strategies used to express these proteins. We show here that the yeast transposon produces a fusion protein by a specific frameshifting event that fuses two out-of-phase open reading frames (ORFs). The process is remarkably similar to that used by retroviruses such as Rous sarcoma virus (RSV) to produce Pr180gag-pol. PMID- 2982103 TI - Where now with AIDS? PMID- 2982104 TI - Nucleic acid structure and expression of the human AIDS/lymphadenopathy retrovirus. AB - The 9,213-nucleotide structure of the AIDS/lymphadenopathy virus has been determined from molecular clones representing the integrated provirus and viral RNA. The sequence reveals that the virus is highly polymorphic and lacks significant nucleotide homology with type C retroviruses characterized previously. Together with an analysis of the two major viral subgenomic RNAs, these studies establish the coding frames for the gag, pol and env genes and predict the expression of a novel gene at the 3' end of the genome unrelated to the X genes of HTLV-1 and -II. PMID- 2982105 TI - A trans-acting factor is responsible for the simian virus 40 enhancer activity in vitro. AB - Stimulation of in vitro transcription by the simian virus 40 enhancer involves a rapid and stable binding of a trans-acting factor with both the 5'- and 3' domains of the enhancer sequence. The enhancer factor, which differs from other types of transcriptional factors, can interact with other enhancer elements. PMID- 2982106 TI - An N-methylaspartate receptor-mediated synapse in rat cerebral cortex: a site of action of ketamine? AB - It has been proposed that three major receptor subtypes subserve the putative transmitter role of glutamate and aspartate in the mammalian central nervous system. One subtype is classified by the specific agonist N-methylaspartate (NMA) and the specific antagonist 4-amino-2-phosphonovaleric acid. It has been shown recently that excitation of neurones by NMA is also selectively reduced by dissociative anaesthetics such as ketamine and phencyclidine and by sigma opiates, drugs of abuse with common psychotomimetic properties. Responses to NMA have an unusual voltage relation which may result from a voltage-dependent block of the activated channel by physiological concentrations of magnesium. No synaptic potential with properties similar to those of responses to NMA, however, has yet been reported. We describe here an excitatory postsynaptic potential (e.p.s.p.) evoked by electrical stimulation of the white matter and recorded intracellularly from pyramidal cells in slices of rat somatosensory cortex. This e.p.s.p. has the appropriate voltage relation and sensitivity to Mg2+ and ketamine to be an NMA receptor-mediated synapse and a potential central site for the psychotomimetic actions of ketamine. PMID- 2982107 TI - Binding sites for growth hormone releasing factor on rat anterior pituitary cells. AB - Growth hormone releasing factors (GRFs) have been isolated from human pancreatic tumours (hGRF) and rat hypothalamus (rhGRF). The response to GRF at the pituitary level can be modulated by other factors, including glucocorticoids, thyroid hormones, somatostatin and other neuropeptides and somatomedins. Glucocorticoids enhance GRF-induced growth hormone (GH) secretion in primary cultures of rat anterior pituitary cells, and the synthetic glucocorticoid dexamethasone has recently been shown to increase the amounts of GH released in freely moving rats in response to submaximal doses of intravenous GRF. To investigate whether somatotroph sensitivity to GRF is modulated at its receptor level, we have developed a radioreceptor assay using an iodinated analogue of hGRF as radioligand. We report here that the relative binding affinities of rGRF, hGRF and the two analogues are correlated with their in vitro biological potencies. Further, the number of GRF binding sites is drastically decreased in cells deprived of glucocorticoids either in vivo or in vitro. PMID- 2982108 TI - Cyclic GMP can increase rod outer-segment light-sensitive current 10-fold without delay of excitation. AB - To test the hypothesis that cyclic GMP is the internal messenger coupling rhodopsin activation to membrane excitation in vertebrate rod photoreceptors, we used a novel technique combining measurement of membrane currents of isolated salamander rods with a suction electrode and the introduction of cyclic GMP through a whole-cell recording patch pipette. Rupture of an attached patch was followed by a rapid (approximately 10 s), approximately 10-fold increase in outer segment membrane current, all of which was light-sensitive. There was little change in the rising phase of the response to a saturating flash, but the duration of the saturated phase of the response increased approximately 10-fold. The effects reversed completely within 3-4 min after withdrawal of the cyclic GMP containing patch pipette. A formal kinetic analysis shows that the first two observations are inconsistent with the postulate that cyclic GMP opens the light sensitive conductance by simple binding to channels, unless free cyclic GMP in the outer segment is assumed to be much lower than published estimates, and most of the outer-segment cyclic GMP is bound and inexchangeable on the timescale of 200 ms. Furthermore, our results suggest that rod cyclic GMP is not involved solely in keeping the light-sensitive conductance open, but may also affect the activity of the phosphodiesterase that mediates cyclic GMP hydrolysis. PMID- 2982109 TI - Autonomous chaotic behaviour of the slime mould Dictyostelium discoideum predicted by a model for cyclic AMP signalling. AB - How sustained oscillations lose their periodicity and thus give rise to chaos was first analysed in mathematical models, then observed in chemical systems such as the Belousov-Zhabotinsky reaction where chaos is autonomous because it originates from endogenous kinetic mechanisms. In contrast, chaos can also be obtained by periodically forcing an oscillatory system, as shown, for example, in cardiac cells and yeast glycolysis. Biochemical evidence for autonomous chaos has been obtained both in vitro for the peroxidase reaction and in enzymatic models not based directly on experimental systems. We report here the occurrence of autonomous chaos in a realistic model for the cyclic AMP signalling system of the slime mould Dictyostelium discoideum, based on receptor modification. This model is also capable of bursting, a phenomenon characteristic of some pacemaker neurones such as R15 in Aplysia. Whereas bursting has not been observed in D. discoideum, our model suggests that 'aperiodic signalling' in the mutant Fr17 provides the first example of autonomous chaos occurring spontaneously at the cellular level. PMID- 2982110 TI - Chronic morphine treatment induces hypersensitivity to testosterone-negative feedback in castrated male rats. AB - Studies were undertaken to determine the effects of chronic stimulation of opiate receptors on the negative feedback effects of testosterone (T) on luteinizing hormone (LH) secretion in the male rat. In an initial study, castrated male rats received replacement levels of T (2 ng/ml) or chronic morphine (M) treatment for 7 days. When initiated at the time of castration, both T and M treatments prevented the castration-induced hypersecretion of LH. However, when the treatments commenced 2 weeks after castration, only T restored LH secretion to the low levels seen in intact rats. In a second study, rats castrated 2 weeks previously were exposed to chronic M or placebo (control) treatment in the presence of various dosages of T. In rats receiving T alone, LH secretion was unaffected at T levels up to 600 pg/ml serum, but thereafter there was a dose dependent suppression of LH release by T. Serum T levels which reduced LH secretion by 50% were estimated to be 966 pg/ml. In contrast, in castrated rats receiving both M and T treatment, a 50% reduction in LH secretion was estimated to be at 300 pg T/ml serum and maximal inhibition of LH secretion was achieved at serum T levels of greater than 600 pg/ml. Neither T alone nor M plus T treatment altered the responsiveness of the anterior pituitary to LHRH in vitro. These findings indicate that M may enhance the sensitivity of the hypothalamus to T feedback by approximately 3-fold and raise the possibility of the existence of an opioid-sensitive neural component which may modulate the negative feedback effects of T on LH secretion. PMID- 2982111 TI - Beta-adrenergic stimulation of adenosine-3',5'-monophosphate (c-AMP) accumulation and of prolactin and growth hormone secretion in rat anterior pituitary cell cultures. AB - The beta-adrenergic agonist 1-isoproterenol (ISO) (10(-9)-10(-7) M) provoked a prompt and profound increase of intracellular c-AMP accumulation in monolayer cultures of rat anterior pituitary. In superfused reaggregate cell cultures ISO also stimulated c-AMP outflow. The effect was blocked by propranolol and the highly selective beta 2-receptor blocker ICI 118.551. Epinephrine (E), norepinephrine (NE) and the highly selective beta 2-agonist zinterol (ZIN) also stimulated c-AMP accumulation. The order of potency ZIN greater than or equal to ISO greater than E much greater than NE together with the high potency of ICI 118.551 suggests the beta-effect is mainly through the beta 2-receptor subtype. The same concentrations of ISO strongly stimulated growth hormone (GH) release and, as previously shown, prolactin (PRL) release from superfused reaggregate cell cultures, but not luteinizing hormone or thyroid stimulating hormone release. Stimulation of PRL and GH release from these cultures was also induced by the adenylyl cyclase activator forskolin. Since interference of beta adrenergic effects on contaminating fibroblasts or endothelial cells could be reasonably excluded, the present data suggest that beta-adrenergic stimulation of c-AMP accumulation in anterior pituitary cells elicits GH and PRL release. PMID- 2982112 TI - Beta-adrenergic stimulation of adenosine-3',5'-monophosphate (c-AMP) in primary cultures of rat anterior pituitary cell populations separated by unit gravity sedimentation. Relationship to growth hormone and prolactin release and to nonsecreting cells. AB - The effect of the beta-adrenergic agonist isoproterenol (ISO) on c-AMP accumulation and on growth hormone (GH) and prolactin (PRL) release was studied in primary cultures of anterior pituitary cell populations with different proportional number of somatotrophs and lactotrophs, obtained by velocity sedimentation at unit gravity. ISO stimulated c-AMP levels in highly-enriched lactotroph (approximately 70%) as well as in highly-enriched somatotroph (approximately 65%) populations. ISO-stimulated c-AMP accumulation was attenuated by dopamine (DA) in highly enriched lactotroph but not somatotroph populations. In a small-cell population consisting of a much lower proportional number of lactotrophs and somatotrophs, the proportional increase of c-AMP accumulation was several times higher than in the other populations and the effect was not suppressible by DA. In this small-cell population, GH but not PRL release was more responsive to ISO than in the other populations. The present observations are consistent with the interpretation that the changes in c-AMP levels induced by ISO originate in part in somatotrophs and lactotrophs. However, the present data also demonstrate that the most responsive cell population in terms of c-AMP accumulation consists of small cells. The majority of these cells could not be identified. The possibility that these cells may be nonsecreting folliculostellate cells is discussed. PMID- 2982113 TI - Attempts to implicate viruses in myasthenia gravis. AB - We investigated the hypothesis that a persistent viral infection of the thymus gland might trigger the autoimmune disease myasthenia gravis (MG). Thymus glands of nine patients with recent onset of MG were studied by a variety of techniques to detect the presence of occult viruses. No evidence of viral infection was found. PMID- 2982114 TI - Myeloneuropathy, neurologic toxicity, and amiodarone. PMID- 2982115 TI - Single dose sulbactam/ampicillin for uncomplicated urinary tract infections. PMID- 2982116 TI - Endometritis and acute salpingitis associated with Chlamydia trachomatis and herpes simplex virus type two. AB - A patient who had endometritis confirmed by endometrial biopsy and acute salpingitis confirmed by laparoscopy is reported. Chlamydia trachomatis and herpes simplex virus type two were isolated from the endometrial cavity and the fallopian tube. The histopathologic findings of the endometritis were similar to those frequently seen in chronic chlamydial eye disease or chlamydial cervicitis. PMID- 2982117 TI - Is porphyria cutanea tarda a risk factor in the development of hepatocellular carcinoma? A case report and review of the literature. AB - A patient with alcoholic liver cirrhosis, diabetes mellitus and porphyria cutanea tarda (PCT) is described, who subsequently died of hepatocellular carcinoma. The literature relating PCT to the incidence of primary hepatoma is reviewed, and the mechanisms underlying this possible association are discussed. PMID- 2982118 TI - Experimentally produced synovial sarcoma in mice. AB - BALB/c and C57 B1/6 mice which received weekly subcutaneous injections of a cannabinoid developed tumors at the points of injection. In the groin the tumors were fibrosarcomas, while in the interscapular region they were anaplastic sarcomas and synovial sarcomas. The latter developed faster and were larger in cannabinol - and cannabidiol-treated C57 B1/6 mice than in delta 9 tetrahydrocannabinol-treated BALB/c mice. This difference could not be attributed to strain differences. 2 cannabidiol- and 1 cannabinol-treated C57 B1/6 females with the fastest growing synovial sarcomas were pregnant and, in addition, 2 of them developed mammary adenocarcinomas, indicating that they were rich in estrogen. On the other hand, it is known that delta 9-tetrahydrocannabinol causes a decrease in the sex hormone. It is consequently thought that the cannabinoids are only initiators of synovial sarcoma, while growth or promotion of the latter is influenced partially or entirely by the sex hormone. PMID- 2982120 TI - [Pathomechanism of hyperthyroidism and hormone receptors]. PMID- 2982119 TI - [A new radioisotope method for the study of placental circulation in intrauterine growth retardation]. PMID- 2982121 TI - [Diagnostic antibody tests for the detection of central nervous system diseases caused by herpes simplex and varicella zoster viruses]. PMID- 2982122 TI - [Wilms tumor in childhood diagnosed during therapy of acute lymphoid leukemia]. PMID- 2982123 TI - Sonographic appearance of cytomegalovirus nephritis in a neonate. AB - In adults and children increased renal cortical echogenicity on ultrasound examination is now well recognized as a feature of renal parenchymal disease due to a variety of etiologies. The degree of echogenicity appears related to the severity of the renal disease, histologically. The sonographic renal pattern of the neonate and in particular, of the premature, differs generally from that of the adult in that the renal cortex of the very young may be "normally" more echogenic than in later life. Marked cortical echogenicity however, is a sign of renal parenchymal disease even in the premature, as illustrated in the case that follows with documented cytomegalovirus nephritis. PMID- 2982124 TI - New tools for the study of Niemann-Pick disease: analogues of natural substrate and Epstein-Barr virus-transformed lymphoid cell lines. AB - Acid sphingomyelinase activity was determined in Epstein-Barr virus-transformed lymphoid cell lines (LCL) established from patients affected with Niemann-Pick disease (NPD) using several substrates: sphingomyelin derivatives, radiolabeled [14C]sphingomyelin (SM), fluorescent N-(10-(1-pyrene)decanoyl)sphingomyelin (P10 SM) or colored trinitrophenylaminolauryl-sphingomyelin, and the chromogenic non natural substrate 2-N-(hexadecanoyl)amino-4-nitrophenylphosphoryl-choline. LCL from NPD Type A and Type B showed a severe deficiency of acid sphingomyelinase determined using either substrate, whereas LCL from normal subjects had an activity close to that of blood leukocytes. Sphingomyelinase in normal LCL had the same pH optimum (5.0-5.2) and molecular form (pI 5.8) as the enzyme from other sources; identical profiles and activity levels were obtained using the various analogues of sphingomyelin. However, among these derivatives, the assay using P10-SM appeared as the most useful and sensitive for enzymatic diagnosis of NPD. Electron microscopy of NPD LCL demonstrated the lysosomal storage. These results prove the validity of LCL as an experimental model system for NPD. PMID- 2982126 TI - Adenylate cyclase activity in fetal rabbit myocardium. AB - Adenylate cyclase activity [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] was determined in vitro in fetal rabbit myocardial membranes from individual fetal pups at 21 to 31 days gestation (term, 31 days). Basal and NaF-stimulated adenylate cyclase activities did not change during 21-31 days gestation. Significant stimulation of the enzyme by l-isoproterenol was observed only in the presence of guanosine triphosphate (100 microM). Under these conditions, maximal adenylate cyclase stimulation by l-isoproterenol (100 microM) was significantly higher at 25-31 than at 21 days gestation. Moreover, EC50 (Kact) for l isoproterenol at 25-31 days was significantly lower than at 21 days gestation. We conclude that, in fetal rabbit myocardial membranes, there is an increase in the sensitivity of adenylate cyclase stimulation by l-isoproterenol from 21 to 25-31 days gestation. PMID- 2982125 TI - Age-related red cell enzymes in children with transient erythroblastopenia of childhood and with hemolytic anemia. AB - Red cell enzymes of three children with transient erythroblastopenia of childhood were measured and compared with those of age-matched normal children and children with hemolytic anemia. While the activity of "age-dependent" enzyme such as hexokinase, aldolase, glucose-6-phosphate dehydrogenase, glutamic-oxaloacetic transaminase, and pyruvate kinase were greatly increased in the red cells of children with hemolytic anemia, they were not decreased in the red cells of children with erythroblastopenia of childhood. Only the activity of pyrimidine 5' nucleotidase was consistently low red cells of these children. These findings are inconsistent with the usual concept that red cell enzyme activities decline throughout red cell life span. Rather, they suggest that there may be very rapid loss in the activity of some red cell enzymes during the first few days of red cell life with little further decline in enzyme activity. PMID- 2982127 TI - Ontogeny of hepatic bile acid conjugation in the rat. AB - We studied the postnatal development of hepatic bile acid conjugation in the rat. Overall conjugating activity (homogenate assay) and the activity of the individual enzymes involved in the two step conjugation reaction (coenzyme A-bile acid thioester formation by microsomal cholyl-CoA ligase and amino acid transfer to this intermediate mediated by cytosolic cholyl-CoA:taurine N-acyltransferase were measured on days 1, 7, 14, 21, and 56 of life. Cholic acid conjugation was significantly lower in the suckling rat on days 1, 7, and 14 compared to the adult rat on day 56. At weaning (21 days) there was a marked increase in homogenate activity to a level higher than at 56 days. This peak in conjugating activity could be precociously induced on day 14 by cortisone acetate injection on days 10 through 13. Both cholyl-CoA ligase and cholyl-CoA:taurine N acyltransferase activities were significantly lower during the first 14 days of life in comparison with 56 days. At 21 days of age there was a sharp increase in cholyl-CoA ligase activity to 1.5 times the activity in the adult; however, cholyl-CoA:taurine N-acyltransferase activity remained less than half of that found in the adult. We conclude that 1) cholic acid conjugation, as assessed by determination of either total conjugating activity or by the individual enzymatic steps, is decreased in the suckling rat; 2) the peak in conjugating activity noted at weaning may be mediated by the hormonal milieu, and 3) cholyl-CoA ligase activity closely parallels the increase in total activity and appears to be rate limiting during development. PMID- 2982128 TI - Noninvasive monitoring of cerebral oxygenation in preterm infants: preliminary observations. AB - A noninvasive optical method for bedside monitoring of cerebral oxygenation in small preterm infants was evaluated. Through differential absorbance of near infrared light, changes in the oxidation-reduction level of cytochrome aa3, in the oxygenation state of hemoglobin and in tissue blood volume were assessed in the transilluminated anterior cerebral field. Overall, cerebral oxygenated hemoglobin correlated significantly with transcutaneous oxygen, r = .44 p less than .0001; however, correlation was best in the absence of cardiorespiratory disease. Hypoxia with or without bradycardia led to hemoglobin deoxygenation and a shift in cytochrome aa3 to a more reduced state. When hypoxic episodes came in series or were prolonged, aa3 reduction occurred simultaneous with hemoglobin deoxygenation but its recovery to base-line values sometimes lagged behind the return of hemoglobin oxygenation. In one infant with a large patent ductus arteriosus, even brief episodes of mild bradycardia caused precipitous reduction of cytochrome aa3 before any shift to greater hemoglobin deoxygenation. This response disappeared after ductal ligation. In general, the antecedent state of cerebral oxygenation, the severity and duration of deoxygenation, and the presence or absence of circulatory abnormalities all influenced the aa3 response to hypoxia. Continuous noninvasive near infrared monitoring of cerebral oxygenation can be performed on sick preterm infants at the bedside. PMID- 2982129 TI - Cytomegalovirus-associated hemophagocytic syndrome. PMID- 2982130 TI - Knowledge and attitudes of persons with cancer toward use of unproven treatment methods. PMID- 2982131 TI - Kawasaki disease associated with Klebsiella pneumoniae bacteremia and parainfluenza type 3 virus infection. PMID- 2982132 TI - Maternal and neonatal neutralizing antibody titers to selected enteroviruses. AB - Neutralizing antibody titers were determined at term for 98 matched mothers' serum and neonatal cord blood pairs, to coxsackieviruses B2, B3, B4 and B5 and to echovirus 11. The reciprocal of the geometric mean titers of maternal sera were, respectively, 15.3, 18.1, 53.5, 14.1 and 11.4. The geometric mean titers of cord sera were statistically lower for coxsackie B5 and echovirus 11 (P less than 0.031 and p less than 0.005, respectively) than those in maternal sera. The percentage of neonates born with antibody titers less than 1:10 to coxsackieviruses B2, B3, B4 and B5 and echovirus 11 ere 49, 37, 17, 53 and 69%. Thus a large percentage of neonates in our community are born with low titers of enterovirus-neutralizing antibody, which for some infants may be disproportionately lower than maternal titers to selected enteroviruses. Commercially available immune serum globulin contains high titers of antibody to these enteroviruses and may be a useful means of passive antibody acquisition in susceptible neonates at risk for enterovirus infections. However, a single dose of immune serum globulin intramuscularly produced little change in circulating neutralizing antibodies to coxsackie B4 and echovirus 11 in seven neonates. PMID- 2982133 TI - Gastrointestinal adenovirus: an important cause of morbidity in patients with necrotizing enterocolitis and gastrointestinal surgery. AB - Adenoviruses have recently been recognized as etiologic agents of enteric disease in hospitalized infants and young children. Patients who have undergone surgical procedures related to the complications of neonatal necrotizing enterocolitis constitute a population of infants who are often hospitalized for extended periods of time. We studied the incidence of enteric adenovirus infections in infants with necrotizing enterocolitis who had undergone intestinal surgery resulting in colostomies or ileostomies. These studies revealed that 31.7% of the hospital admissions in such patients were complicated by symptomatic infection with enteric adenoviruses. The rate of adenovirus infection in this population was significantly higher than the rate of such infections encountered in infants undergoing ostomy procedures for other conditions and in age-matched hospitalized infants without prior gastrointestinal surgery. The infections with enteric adenovirus were generally accompanied by an increase in intestinal output which was temporally associated with the excretion of adenovirus in the intestinal fluid. In addition infections with enteric adenoviruses in the infants with necrotizing enterocolitis and ostomies were associated with a marked increase in length of hospitalization as compared to uninfected infants with similar underlying gastrointestinal pathology. Epidemiologic analysis revealed that many of the cases of enteric adenovirus infection in the study population occurred during seasons in which enteric adenovirus infections were prevalent in the hospital population. These studies demonstrate that enteric adenovirus infections are major causes of morbidity in hospitalized patients who have of morbidity in hospitalized patients who have undergone ileostomy or colostomy procedures for necrotizing enterocolitis and that the prevention of enteric adenovirus infections might result in a significant improvement in the hospital care of infants with these conditions. PMID- 2982134 TI - Chloramphenicol toxicity: hemodynamic and oxygen utilization effects. PMID- 2982135 TI - Hemorrhoids and anal fissures. Common problems, current solutions. AB - Recent insights into the nature of hemorrhoids and anal fissures have led to specific strategies for treatment. Symptomatic hemorrhoids often result from prolapse of submucosal vascular cushions. Appropriate treatments include dietary change, injection therapy, rubber-band ligation, and hemorrhoidectomy. Patients with anal fissures usually have an abnormal contraction of the sphincter. Many fissures heal spontaneously. Treatment often centers on helping the patient avoid constipation, although surgery is indicated in some cases. PMID- 2982136 TI - [Non bacterial thrombotic endocarditis associated with a latent primary hepatoma]. PMID- 2982137 TI - [Quick diagnosis of eye infections caused by adenoviruses]. PMID- 2982138 TI - Reexamination of the evidence for paramagnetism in oxy- and carbonmonoxyhemoglobins. AB - Experiments have been carried out in an attempt to confirm previous reports of paramagnetism in the oxy- and carbonmonoxy derivatives of human and carp hemoglobin. When care is taken to ensure complete saturation of the hemoglobins with ligand and the diamagnetic contributions of all of the buffers are carefully evaluated, these hemoglobin derivatives are found to have the same gram susceptibilities as exhibited by a set of metal free proteins, suggesting that they are fully diamagnetic. PMID- 2982139 TI - Accumulation of 10-kilobase DNA replication intermediates in cells treated with 3 aminobenzamide. AB - During eukaryotic DNA synthesis there is formation of, in addition to Okazaki fragments, discrete 10-kilobase (kb) DNA replication intermediates. We have investigated the ligation of 10-kb DNA replication intermediates to high molecular weight DNA, using the drug 3-aminobenzamide, an inhibitor of poly(ADP ribose) synthetase. In human melanoma cells treated with this inhibitor, there is an accumulation of 10-kb DNA. In contrast, in cells treated with aphidicolin, which inhibits DNA polymerase alpha, there is continued ligation of 10-kb DNA to high molecular weight DNA. Furthermore, using sequential treatment with aphidicolin and 3-aminobenzamide, one can observe the conversion of radiolabeled Okazaki fragments into 10-kb intermediates. The 10-kb DNA pieces are, however, not ligated to high molecular weight DNA in the presence of 3-aminobenzamide. Our results imply that functioning poly(ADP-ribose) synthetase is necessary for the ligation process. PMID- 2982140 TI - Genetic analysis of beta-adrenergic receptor internalization and down-regulation. AB - We have used wild-type and variants of the T-lymphoma cell line S49 to explore internalization and down-regulation of adenylate cyclase-linked beta-adrenergic receptors. Internalization was defined by the loss in "surface receptors" detected at 4 degrees C on intact cells by the antagonists [3H]CGP-12177 or [125I]iodocyanopindolol, whereas down-regulation was defined as the loss in total cellular content of receptors [( 125I]iodocyanopindolol binding assayed at 37 degrees C). In wild-type cells, the beta-adrenergic agonist isoproterenol induced a rapid (t 1/2, approximately equal to 1 min) and reversible loss in surface receptors. The surface sites were lost at a rate similar to the rate of desensitization of beta-adrenergic receptor-mediated cyclic AMP generation of S49 cells. A series of S49 variants (cyc-, UNC, H21a) having lesions in NS (the guanine nucleotide binding protein that couples beta-receptors to adenylate cyclase) or with absent cAMP-dependent protein kinase activity (kin-), had a loss in surface sites that was equivalent to that of wild-type cells. By contrast, S49 variant cells having lesions in NS showed variable rates and extents of down regulation of beta-adrenergic receptors. In wild-type and kin- S49 cells, beta receptors down-regulated with a t 1/2 of approximately equal to 4 hr. Down regulation was blunted in the cyc- and UNC variants that have altered coupling of receptors to NS, but it was faster in the H21a variant that retains receptor-NS interaction. Recovery of receptors after down-regulation occurred at a similar rate (t 1/2, approximately equal to 6 hr) in wild-type, UNC, and H21a cells. These results demonstrate that internalization of beta-adrenergic receptors may be necessary, but is not sufficient, to explain agonist-induced receptor down regulation in S49 cells. The variable expression in the development of down regulation in S49 variants implies that receptor-NS interaction regulates the fate of receptors linked to the stimulation of adenylate cyclase. PMID- 2982141 TI - Transfer of genes into hematopoietic cells using recombinant DNA viruses. AB - The ability of recombinant DNA viruses to transfer genes into hematopoietic cells has been explored. A recombinant simian virus 40 (SV40) in which the early region had been replaced with the chloramphenicol acetyltransferase (CAT) gene driven by the promoter from Rous sarcoma virus (RSV), was constructed. This virus transferred the CAT gene more efficiently into mouse and human bone marrow cells and into the K562, MEL, and WEHI hematopoietic tissue culture cell lines, than the classical calcium phosphate DNA transfer procedure, as shown by assay for CAT activity 48 hr after infection. Recombinant SV40 virions were also shown to be capable of stably transforming Chinese hamster ovary cells by use of an early region recombinant containing the methotrexate-resistant dihydrofolate reductase (DHFR) gene driven by the RSV promoter. The entire DHFR transcriptional unit could be detected in the genome of transformed cells that were also shown to be resistant to methotrexate. A recombinant adenovirus stock containing the neomycin resistance gene driven by the SV40 early promoter was used to infect the K562 and MEL hematopoietic cell lines to resistance to the antibiotic G418. Transformation frequency was 10- to 100-fold higher than that obtained with calcium phosphate precipitated DNA. Most or all of the recombinant adenovirus genome was integrated as 1-3 copies in the transformed cells. These studies show the feasibility of using DNA viruses for introduction of new genetic material into hematopoietic cells. PMID- 2982142 TI - Mutations that affect the translation efficiency of Tn9-derived cat gene in Bacillus subtilis. AB - We have isolated two spontaneous mutations that increase the expression of the Tn9-derived cat gene in Bacillus subtilis. These mutations, which appear to affect initiation of translation of chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) consist of a tandem duplication and triplication of a 55-base-pair sequence located at the 5' end of cat. Included in the repeated sequence are the Shine-Dalgarno site, initiation codon, and a region of dyad symmetry located within the structural portion of the cat gene. A striking feature of the mutated initiation sites is their potential to form stem-loop structures at the 5' end of the cat messenger RNA. Within the single-stranded loops of these structures are the ribosome binding site and initiation codon for the cat gene. It appears that the Gram negative cat translation initiation site has mutated to permit efficient utilization in B. subtilis without directly affecting Shine-Dalgarno sequence homology. This report suggests that secondary structure in the vicinity of the Shine-Dalgarno site can exert a strong positive influence on the initiation of translation in B. subtilis. PMID- 2982144 TI - Formation of a transformed follicle is necessary but not sufficient for development of an avian leukosis virus-induced lymphoma. AB - Avian leukosis virus (ALV) infection of susceptible chickens induces bursal lymphomas after a latent period of several months. The clonal development of these B-cell tumors is believed to be a multistep process. Histopathological changes, referred to as transformed follicles, occur within the target organ soon after virus infection and may represent a proximal stage of lymphomagenesis. To establish further the significance of this lesion and its relationship to the subsequent development of lymphomas, we have compared the incidence of transformed follicles observed in animals susceptible or resistant to ALV-induced tumor development. During the 8 weeks following ALV infection, transformed follicles were detected in 82% of the susceptible animals and in 11% of the resistant animals. These results indicate that the incidence of transformed follicles in these animals correlates with their susceptibility to lymphoma development. Furthermore, each transformed follicle does not develop into a tumor. These observations suggest that the formation of a transformed follicle is necessary but not sufficient for lymphoma development. PMID- 2982143 TI - Inversely repeating integrated hepatitis B virus DNA and cellular flanking sequences in the human hepatoma-derived cell line huSP. AB - Among recombinant phages carrying integrated hepatitis B virus (HBV) DNA sequences cloned from the human hepatoma-derived cell line huSP, one clone, lambda hu-489, revealed some unusual features. The 2.25-kilobase Eco D fragment from the insert of this clone hybridized to the HBV DNA probe only and its nucleotide sequence was determined. The viral sequence, as well as a cellular flanking sequence, showed extensive rearrangement accompanied by inverted repetition. The Eco D fragment contained HBV DNA from the 5'-end region of gene S to the middle of gene X, followed by a long cellular flanking sequence. Moreover, a part of gene X was found inversely repeated at the head of the same gene S in a head-to-head configuration truncated by the same cellular sequence. Therefore, the same junction sequence of viral DNA and the cellular sequence was found at two different sites in the Eco D fragment in opposite polarities. PMID- 2982145 TI - Identification and partial characterization of a prolactin-like hormone produced by rat decidual tissue. AB - Previous studies have strongly, but indirectly, suggested that rat decidual tissue produces a prolactin-like hormone, decidual luteotropin, which markedly affects luteal cell function. However, it was also found that extracts of decidual tissue do not cross-react with antisera to either rat or ovine prolactin (PRL). The purpose of this study was to determine whether the decidual tissue contains a substance that binds to PRL receptors in rat luteal membranes and, if so, to identify, quantitate, and characterize this molecule with the use of an ovarian radioreceptor assay. Decidual tissue was induced in day 5 pseudopregnant rats by scratching the antimesometrial wall of the uterus; it was collected on day 9 and homogenized and extracted. Decidual tissue extracts bound specifically to ovarian PRL receptors. Graded dilutions of the extracts yielded curves that were parallel to the ovine PRL standard, indicating that decidual luteotropin competes for the same receptor sites on rat luteal membranes. To determine the levels of decidual luteotropin throughout pseudopregnancy, decidual tissue was obtained on each day between days 6-12. The PRL-like activity was detectable in decidual tissue as early as day 6, reached a maximum on day 9, and declined thereafter. The elution profile obtained from gel filtration of a day 9 decidual tissue extract displayed a major component of decidual luteotropin eluting at a Ve/Vo ratio of approximately equal to 2.0. Column chromatography indicated that decidual luteotropin corresponds to a protein with a molecular weight of 23,500. The hormone was heat labile, digestible by trypsin, and appears to contain disulfide linkages. In summary, this study reports the identification, quantitation, and partial characterization of a PRL-like hormone produced by the decidual tissue of the rat. PMID- 2982147 TI - On the mechanism of renaturation of complementary DNA strands by the recA protein of Escherichia coli. AB - The renaturation of complementary DNA strands by the recA protein of Escherichia coli has been found to exhibit the following features. (i) Optimal renaturation occurs at recA protein levels below that required to saturate the DNA strands; saturating amounts of recA protein significantly reduce the rate of reaction. (ii) The reaction proceeds in the absence of a nucleotide cofactor but is markedly stimulated by ATP in the presence of 10 mM Mg2+. A similar stimulation occurs in the absence of ATP when the Mg2+ concentration is increased from 10 mM to 30-40 mM. (iii) Both the ATP-stimulated and the Mg2+-stimulated reactions follow apparent first-order kinetics. These results, taken together with the known effects of ATP and Mg2+ on the state of aggregation of recA protein, suggest that the association of recA monomers may play an important role in recA protein-promoted DNA renaturation. PMID- 2982146 TI - Hybrid hepatitis B virus-host transcripts in a human hepatoma cell. AB - The human PLC/PRF/5 hepatoma cell line (the Alexander cell) contains at least seven copies of hepatitis B virus (HBV) DNA integrated in its genome; but it selectively expresses the HBV surface antigen (HBsAg) gene and perhaps low levels of the core gene. We have prepared a cDNA library from PLC/PRF/5 cell poly(A)+ RNA and isolated clones containing HBV sequences. Hybridization experiments show that the great majority of HBV-specific RNAs in this cell line contain HBsAg coding sequences and are presumably derived from the HBsAg gene. Primer extension experiments show that these HBsAg mRNAs are, however, derived from multiple initiation sites in the HBsAg gene and involve two promoters: one at the 5' end of the gene that can produce a protein of 45 kDa, and one located in the pre-S region that can produce two proteins of 31 kDa and the mature HBsAg, 25 kDa, respectively. The HBV RNAs are hybrid RNA species that contain HBV sequences at their 5' ends and host DNA sequences at the 3' ends. The great majority of these hybrid RNAs are transcribed from two closely related yet distinct HBV integrants. The viral-host sequences of these two related hybrid RNAs suggest that the related HBV sequences were generated from a parental fragment via duplication, translocation, and mutagenesis. These processes may play a role in HBV-related oncogenesis. PMID- 2982148 TI - Identification of new gene products coded from X regions of human T-cell leukemia viruses. AB - Antibodies were raised against oligopeptides deduced from the nucleotide sequence in the conserved region located between env and the 3' long terminal repeat in human T-cell leukemia virus type I (HTLV-I) and type II (HTLV-II) to detect a protein coded from this region in virus-infected cells. Two of these antibodies precipitated a protein of 41 kilo-daltons in HTLV-I-infected cell lines and a protein of 38 kilo-daltons in HTLV-II-infected cells. The protein in HTLV-I infected cells was precipitated by plasma from patients with adult T-cell leukemia but not by plasma from a normal adult. These results indicate that these proteins were translated from new coding regions (X) present in HTLV-I and HTLV II. PMID- 2982150 TI - Phosphorylation enzymes of the propionic acid bacteria and the roles of ATP inorganic pyrophosphate, and polyphosphates. AB - It is shown that polyphosphates are not generated in significant amounts in the phosphoglycerate kinase reaction; polyphosphate is more effective than ATP in the formation of glucose 6-P by glucokinase, but the rate with ATP may be adequate to meet the requirements of glucose metabolism; PPi is far more effective than ATP as a phosphate donor in the formation of fructose 1,6-P2 by phosphofructokinase; PPi rather than ATP almost certainly is used in this reaction; and, aside from glucokinase and phosphofructokinase, the enzymes of phosphorylation are specific in their requirements of phosphate donors or acceptors and are present in adequate amounts to meet the requirements of glucose metabolism by the propionic acid bacteria. PMID- 2982151 TI - Purification of the Rous sarcoma virus src kinase by casein-agarose and tyrosine agarose affinity chromatography. AB - A simple and effective purification method for the src kinase, the transforming gene product of Rous sarcoma virus, has been developed by using affinity chromatography on casein-agarose and tyrosine-agarose columns. NaDodSO4/polyacrylamide gel electrophoresis and silver staining analysis showed that the purified kinase preparation was composed of a predominant polypeptide of 60,000-Da. In most of the preparations, however, three minor proteins (54,000, 52,000, and 15,000 Da) were also detected, and they were partially characterized. As one of the exogenous substrates, calmodulin was found to be phosphorylated on tyrosine by the purified src kinase. PMID- 2982149 TI - Mechanism of inhibition of DNA gyrase by analogues of nalidixic acid: the target of the drugs is DNA. AB - Norfloxacin is a nalidixic acid analogue and one of the most potent DNA gyrase inhibitors. To study the mechanism of this important class of inhibitors, the binding of [3H]norfloxacin to gyrase and substrate DNA was measured. We found that, contrary to prior belief, norfloxacin does not bind to gyrase but instead binds to DNA. This was demonstrated by both equilibrium dialysis and membrane filtration techniques. Binding to ColE1 and pBR322 plasmids showed a primary process that is saturated at a norfloxacin concentration about equal to its supercoiling Ki (1.8 X 10(-6) M) and is followed by weaker secondary binding. The apparent Kd values are 1 X 10(-6) M for both plasmids. The molar binding ratio at this initial saturation point is extremely low: only 4 X 10(-4) norfloxacin per nucleotide for both plasmids. The binding of norfloxacin to DNA plasmids is nonintercalative, as shown by the fact that the drug binds preferentially to single-stranded DNA rather than to double-stranded DNA. The binding is reduced at high salt concentration, has a pH optimum between 4.5 and 6.5, and does not require divalent ions. The binding affinities of other nalidixic acid analogues were estimated by an indirect competition method. The calculated apparent Kd values of these analogues correlate well with their Ki values, providing strong evidence that the binding affinity of the drug to DNA determines biological potency. PMID- 2982152 TI - Direct cross-linking of 125I-labeled glucagon to its membrane receptor by UV irradiation. AB - 125I-labeled glucagon was directly crosslinked to its receptor in isolated liver plasma membranes and on the surface of intact hepatocytes, by using a UV irradiation procedure. This investigation resulted in the identification of a glucagon-receptor complex of apparent Mr 62,000. The specificity of labeling was shown by the interference of unlabeled hormone at physiological concentration with incorporation of radioactive glucagon into the 62,000 Mr species. The receptor behaved as a typical integral membrane protein: it was not released by extraction with lithium diiodosalicylate or at basic pH but was solubilized by digitonin treatment. Reduction of the receptor polypeptide with dithiothreitol resulted in a decrease in its electrophoretic mobility, suggesting the presence of intramolecular disulfide bonds. Soluble glucagon-receptor complexes adsorbed to Con A-Sepharose and could be eluted with methyl alpha-D-mannoside, indicating that the receptor molecule is a glycoprotein. Treatment of glucagon-labeled liver plasma membrane with endoglycosidase F resulted in the appearance of four intermediate species, indicating that glucagon receptor contains at least four N linked oligosaccharide chains. PMID- 2982153 TI - Priming of human mitochondrial DNA replication occurs at the light-strand promoter. AB - Individual promoters for transcription of each strand of human mtDNA are located near the origin of heavy-strand DNA replication in the displacement-loop region. Initiation of heavy-strand synthesis represents the first event in mtDNA replication. Analyses of the 5' and 3' map positions of displacement-loop nucleic acids from mitochondria of cultured human cells reveal a close correspondence between the 3' ends of RNA, whose 5' ends map at a unique site, and the 5' ends of DNA strands. The 5' ends of the RNA species all map at nucleotide position 407 in the genomic sequence, which corresponds exactly to the major 5' transcriptional start site, determined previously in vitro, that is contained within the light-strand promoter sequence. Displacement-loop heavy-strand DNAs map immediately adjacent to the 3' termini of these RNAs, and these transition points between RNA and DNA lie within short conserved sequence blocks in the template sequence. The simplest interpretation of these data is that replication is initiated at the major transcriptional promoter with subsequent precise cleavage of primary transcripts to provide the appropriate primer species. PMID- 2982154 TI - Ha-ras proteins exhibit GTPase activity: point mutations that activate Ha-ras gene products result in decreased GTPase activity. AB - Several ras genes have been expressed at high levels in Escherichia coli and the resultant ras proteins were shown to be functional with respect to their well known specific, high-affinity, GDP/GTP binding. We were able to detect a weak GTPase activity associated with the purified proteins. The normal cellular ras protein (p21N) exhibits approximately equal to 10 times higher GTPase activity than the "activated" proteins. Even though the turnover rate of the reaction is very low (0.02 mol of GTP hydrolyzed per mol of p21N protein per minute), the reaction appears to be catalytic; one molecule of p21N hydrolyzes more than one molecule of GTP. The GTPase and the GDP binding activities both have been recovered from a Mr 23,000 protein eluted following NaDodSO4/polyacrylamide gel electrophoresis, suggesting that these two activities are associated with the same protein. Mg2+ ions and dithiothreitol are required for GTPase activity and the optimal pH is between 7 and 8. Guanidine X HCl, which is required for solubilizing bacterially expressed ras protein, is strongly inhibitory to GTPase activity at concentrations higher than 0.5 M. PMID- 2982155 TI - Spin-label oximetry: kinetic study of cell respiration using a rapid-passage T1 sensitive electron spin resonance display. AB - An unusual ESR display has been developed that exhibits sensitivity to bimolecular collisions of dissolved oxygen in water with nitroxide radical spin probes at oxygen concentrations as low as 0.1 microM, requiring only 1 microliter of sample. The method involves observation of the ESR rapid-passage signal when tuned to the dispersion using a loop-gap resonator. The bimolecular collision rate determines the phase of the signal. The method has been used in a closed chamber geometry to study respiration of asynchronous populations of Chinese hamster ovary (CHO) cells. An integral of the Michaelis-Menten equation permits direct comparison with experiment and is shown to be incompatible with the data. The theory of diffusion limitation also is developed and shown to be inconsistent with experiment. The average oxygen concentration is found to decrease as Vmaxt, where t is the time after sealing the chamber, to a critical oxygen concentration of 5.2 microM. Below 5.2 microM, the concentration can be fitted to an exponential form, exp(-t/tau), where tau = 15 sec for 4000 cells per microliter. It is believed that this experimental behavior is determined by complex enzyme kinetics. PMID- 2982156 TI - Distinctive effects of the viral oncogenes myc, erb, fps, and src on the differentiation program of quail myogenic cells. AB - The relationship between susceptibility to transformation in vitro by different oncogenes and terminal differentiation was analyzed in embryonic quail myogenic cells. Infection with Rous sarcoma virus (RSV), Fujinami sarcoma virus (FSV), avian erythroblastosis virus (AEV), and the avian myelocytomatosis virus MC29 led to rapid and massive transformation. Transformed cells had distinctive morphological alterations, increased proliferation rates, and the ability to grow in agar suspension. Furthermore, homogeneously transformed cultures failed to fuse into multinucleated myotubes and to express muscle-specific genes. However, cloned populations of RSV-, FSV-, and AEV-transformed myogenic cells could, under appropriate culture conditions, partially differentiate into atypical "revertant" myotubes. In contrast, competence for terminal differentiation was completely and irreversibly suppressed on transformation by MC29. The specificity of action of a given oncogenic sequence on the inhibition of differentiation was further studied by using conditional and nonconditional transformation mutants. Myogenic cells infected with temperature-sensitive (ts) mutants of RSV and FSV exhibited a fully reversible block of differentiation after shift to restrictive temperature, while cells infected with ts34 AEV were not temperature sensitive for differentiation. Cultures infected with the partially transformation-defective mutant of MC29 td10H were morphologically transformed and acquired anchorage independence for proliferation but maintained a residual competence for terminal differentiation. PMID- 2982157 TI - Developmental variations in the splicing pattern of transcripts from the Drosophila gene encoding myosin alkali light chain result in different carboxyl terminal amino acid sequences. AB - The total sequence of the Drosophila melanogaster gene encoding the myosin light chain dissociated by alkali (MLC-ALK) has been determined. By sequence comparisons with an MLC-ALK cDNA clone and by S1 nuclease analyses, the pattern of introns and exons within the gene has been deduced. There are multiple polyadenylylation signals that can account for most of the observed heterogeneity in the lengths of mRNAs. In the 3' half of the gene, there are two alternative splicing patterns which result in mRNAs that translate to give proteins with two alternative 14 amino acid carboxyl-terminal sequences. There is developmental regulation of the selection of the above splicing sites. One splicing pattern produces an mRNA that translates into a protein used for both larval and adult musculature, whereas the other splicing pattern is used for the latter stage only. PMID- 2982158 TI - Hypoxanthine is the principal inhibitor of murine oocyte maturation in a low molecular weight fraction of porcine follicular fluid. AB - Studies were carried out to identify and quantify the porcine follicular fluid (PFF) component(s) responsible for inhibition of murine oocyte maturation. A low molecular weight fraction of PFF (less than 1000) was prepared by dialysis and used in all experiments. This PFF fraction contained an inhibitor(s) of mouse oocyte maturation that absorbed maximally in the ultraviolet (UV) range at 250 260 nm. When the PFF fraction was charcoal-extracted, significant loss of absorbance at 250, 260, and 280 nm resulted, which corresponded to loss of inhibitory activity. Four major components of PFF were separated by ion-exchange chromatography and characterized according to their UV spectral characteristics and inhibitory activity. When individual fractions demonstrating identical spectra were pooled and analyzed by high-performance liquid chromatography, the first pooled fraction (A) was found to be impure, but adenine comprised 80% of the UV-absorbing material. Fractions B, C, and D were characterized as pure uracil, hypoxanthine, and 7-methylinosine, respectively. The concentrations of these compounds in PFF were estimated to be 0.06 mM adenine, 0.44 mM uracil, 1.41 mM hypoxanthine, and 0.19 mM 7-methylinosine. Comparison of the potencies of commercial preparations of these compounds established that hypoxanthine is the major inhibitory component of the low molecular weight PFF fraction. Moreover, a commercial preparation of hypoxanthine mimicked the action of PFF on mouse oocyte maturation in that it produced a transient inhibition of oocyte maturation that was significantly potentiated by follicle-stimulating hormone and dibutyryl cyclic adenosine monophosphate. When the inhibitory efficacies of purine and pyrimidine bases and nucleosides were compared, their relative potencies in decreasing order were purine bases greater than purine nucleosides greater than pyrimidine bases = pyrimidine nucleosides. We conclude that hypoxanthine is the predominant low molecular weight component of PFF that inhibits mouse oocyte maturation but that other purines/pyrimidines may also play a role in vivo in maintaining meiotic arrest. PMID- 2982159 TI - Isolation and analysis of the 21q+ chromosome in the acute myelogenous leukemia 8;21 translocation: evidence that c-mos is not translocated. AB - Acute myelogenous leukemia (AML), subgroup M2, is associated with a nonrandom chromosomal translocation, t(8;21)(q22,q22). The oncogene c-mos also has been localized to the q22 band on chromosome 8. There is also evidence that genes on chromosome 21 may be important in the development of leukemia. To determine whether the c-mos oncogene has been translocated in AML-M2 with this translocation and to isolate DNA sequences and genes from these two chromosomes that may be important in malignancy, we constructed somatic cell hybrids between a Chinese hamster ovary cell (CHO) mutant defective in glycine metabolism and myeloblasts with an 8;21 translocation from a patient with AML. We isolated the 21q+ chromosome of this translocation in a somatic cell hybrid and showed that the c-mos oncogene had not been translocated to chromosome 21, ruling out the possibility that translocation of c-mos to chromosome 21 is necessary for development of AML-M2. In addition, there was no detectable rearrangement of the c-mos locus within a 12.4-kilobase region surrounding the gene, indicating that rearrangement of the coding region of the gene itself or alteration of proximal 5' or 3' flanking sequences is not involved. We used this hybrid to determine whether specific DNA sequences and biochemical markers from chromosomes 8 and 21 had been translocated in this case. PMID- 2982160 TI - Endonuclease III (nth) mutants of Escherichia coli. AB - Two strains that overproduce endonuclease III were found in a colony bank containing hybrid ColE1-Escherichia coli plasmids. The enzyme was identified in crude extracts by the degradation of partially depyrimidinated DNA in the presence of EDTA, by its sedimentation velocity, and by its associated thymine glycol-DNA glycosylase activity. An insertion mutation was produced by cloning the kanamycin-resistance gene of Tn5 into the plasmid copy of the nth gene. The mutation was then transferred to the chromosome in the following steps: (i) selection for chromosomal integration of the plasmid at 42 degrees C in a temperature-sensitive polA strain, (ii) curing via temperature shifts, and (iii) phage P1-mediated transduction of a new host. The insertion mutant, as well as a separately isolated deletion mutant, had no measurable glycosylase activity for DNA containing thymine glycol. Although such residues are common lesions in oxidized or irradiated DNA, the mutants were not unusually sensitive to H2O2 or gamma-rays. The insertion mutation had a mutator effect (4- to 22-fold enhancement) on one tested allele. PMID- 2982162 TI - Genetic control of chromosome length in yeast. AB - The chromosomes of the yeast Saccharomyces cerevisiae terminate with sequences that have the form poly(C1-3-A). In this paper, we show that within an individual yeast strain all chromosomes end with tracts of poly(C1-3-A) of similar lengths; however, different strains can have tracts that vary in length by a factor of two. By a genetic analysis, we demonstrate that yeast cells have a mechanism that allows them to change rapidly the length of their chromosomes by altering the length of the poly(C1-3-A) tract. PMID- 2982161 TI - A synthetic HIS4 regulatory element confers general amino acid control on the cytochrome c gene (CYC1) of yeast. AB - Hybrid promoters constructed from upstream sequences of the yeast HIS4 promoter and the downstream element of the yeast CYC1 promoter place iso-1-cytochrome c (CYC1) expression under the general amino acid control, typical of HIS4. HIS4 fragments that confer regulation contain at least one copy of the sequence T-G-A C-T-C that is repeated at HIS4 and other genes subject to the general control. A 14-base-pair synthetic oligonucleotide containing a single copy of the HIS4 repeat places CYC1 under the general control. Two copies of this oligonucleotide produce a derepressed level of expression nearly equivalent to that conferred by the largest HIS4 5' noncoding fragments we examined and direct regulated expression of a set of transcripts with 5' ends typical of the CYC1 promoter. Comparison of the expression levels conferred by the short synthetic repeat and larger HIS4 5' fragments reveals additional promoter elements required for maintaining efficient gene expression under repressing growth conditions. PMID- 2982163 TI - Immunoglobulin JH, C mu, and C gamma gene rearrangements in human B lymphocytes clonally transformed by Epstein-Barr virus. AB - Somatic rearrangements and deletions of immunoglobulin gene segments have been demonstrated in several types of murine B cells. In addition, rearrangements of the JH, C mu, and light chain immunoglobulin gene segments have been reported in human pre-B-cell leukemias and B-cell lymphomas. We have used recombinant DNA probes for the human JH, C mu, and C gamma immunoglobulin gene loci to analyze the genetic events associated with heavy chain gene expression in human B cells clonally transformed by Epstein-Barr virus. Southern hybridization analysis of BamHI-digested cell clone DNAs shows that these human B-cell clones often have bi allelic JH rearrangements and that heavy chain isotype switching is associated with bichromosomal C mu and C gamma gene rearrangements. Deletions of germ line C mu and C gamma segments were observed that were sometimes bi-allelic. Overall, the observed rearrangements and deletions of heavy chain constant region genes suggest that human heavy chain class switching proceeds in a general order consistent with the proposed order of the heavy chain gene classes along chromosome 14. PMID- 2982164 TI - Purification and characterization of the mu opiate receptor from rat brain using affinity chromatography. AB - Opiate receptors have been solubilized from rat neural membranes and purified 500 fold (relative to the crude solubilized extract) by affinity chromatography. Active receptors were solubilized by using 3-[( 3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a zwitterionic derivative of cholic acid. Affinity chromatography was carried out using Affi-Gel 401, a sulfhydryl derivative of agarose to which "hybromet," a newly synthesized opioid ligand with high affinity for the mu receptor, had been attached. Scatchard analysis of [3H]etorphine binding to the purified receptor revealed a single class of high affinity sites (Kd = 1.4 nM; Bmax = 2800 fmol/mg of protein). Half-maximal binding was achieved at approximately equal to 1 nM. Activity was markedly inhibited by protein modifying reagents, findings which suggest that the sites are proteinaceous. Opiate binding activity was also inhibited by the guanyl nucleotide GTP. Electrophoresis of the purified material under denaturing conditions revealed three subunits of molecular weights 94,000, 44,000, and 35,000. The inhibitory guanyl nucleotide binding protein (Ni) implicated in opiate action has been shown to be comprised of two subunits of molecular weights 42,000 and 35,000. Thus, the opiate receptor may be an aggregate of multiple protein components that may include a guanyl nucleotide binding protein. PMID- 2982165 TI - Characterization of octopamine-sensitive adenylate cyclase: elucidation of a class of potent and selective octopamine-2 receptor agonists with toxic effects in insects. AB - Octopamine-2 receptors, associated with activation of adenylate cyclase, mediate a number of the important hormonal and neurotransmitter functions of octopamine in invertebrates. By utilizing the highly enriched octopamine-sensitive adenylate cyclase present in the firefly light organ, it has been possible to pharmacologically characterize octopamine-2 receptors and to define a new class of highly potent and selective octopamine-2 agonists. At low concentrations, these substituted phenyliminoimidazolidines stimulate light emission when injected into fireflies. At somewhat higher concentrations, these compounds, when ingested by tobacco hornworms, cause disruption of motor and feeding behavior, leading to insect death. The effects of these compounds are markedly potentiated by phosphodiesterase inhibitors and mimicked by other activators of octopamine sensitive adenylate cyclase, including octopamine itself. Because octopamine-2 receptors appear to be present primarily in invertebrates, these findings, together with other data, raise the possibility that potent and selective octopamine agonists could be useful as insect toxins with low toxicity in vertebrates. PMID- 2982166 TI - Transmission at voltage-clamped giant synapse of the squid: evidence for cooperativity of presynaptic calcium action. AB - Synaptic transmission was studied at the squid giant synapse with voltage clamp control of both presynaptic and postsynaptic cells. Because presynaptic voltage gradients can complicate interpretation of electrophysiological data obtained from this preparation, we used local Ca application to restrict Ca influx and transmitter release to a short and relatively isopotential portion of the elongated presynaptic terminal. Under these conditions, we found that postsynaptic current varies approximately as the third power of presynaptic Ca current. This finding is consistent with the hypothesis that several Ca ions cooperate in triggering secretion of a single transmitter quantum. PMID- 2982167 TI - The effects of valproate and phenytoin on the cAMP and cGMP levels in nervous tissue. AB - Earlier studies have demonstrated that valproic acid (VPA) and phenytoin (PHT) influence the excitability properties of crayfish axons through different mechanisms. PHT was found to antagonize the electrophysiologic effects of VPA. The purpose of the present study was to determine if the electrophysiologic effects of VPA and PHT are correlated with changes in the cellular levels of either cAMP or cGMP as these substances are known to influence membrane excitability. It was found that PHT (0.1 mM) has no effect on the levels of either cAMP or cGMP within crayfish neural tissue. VPA (4.0 mM) also has no effect on cAMP levels. However, it does significantly reduce the levels of cGMP. Pretreatment of neural tissue with PHT has been shown to eliminate the effects of VPA on membrane excitability. It was found that this pretreatment has no influence on VPA's ability to reduce cGMP levels. The effect of VPA on cGMP levels is observed in the absence of spontaneous activity. Therefore, it is concluded that the observed reduction in cGMP levels does not represent the modulation of cGMP levels that is known to accompany activity. Two experiments demonstrate that the 4-mV depolarization of membranes by VPA can not account for its effect on cGMP levels. In the first, pretreatment with PHT abolished the depolarizing effect on VPA but not its effect on cGMP. In the second, a concentration of ouabain which depolarizes crayfish neural tissue by 8-10 mV without producing spike activity had no effect on either cAMP or cGMP levels. These experiments effectively dissociate the electrophysiologic response to VPA and PHT from changes in cyclic nucleotide levels. PMID- 2982168 TI - Effects of naloxone on regional blood flow distribution in canine hemorrhagic shock. AB - The opiate antagonist naloxone increases arterial pressure, maximal left ventricular dp/dt and cardiac output when administered to dogs subjected to hemorrhagic shock. The purpose of this study was to investigate regional blood flow changes associated with naloxone treatment in anesthetized hypovolemic and normovolemic dogs. Hypovolemic dogs (n = 10) were bled over 30 min (t = -30 to t = 0) to a pressure of 45 mm Hg which was maintained for 1 hr. At t = 60, five dogs received naloxone (2 mg/kg + 2 mg/kg X hr), and five received an equal volume of saline. Regional blood flows were determined at t = -30, 45, and 90 min using 15-micron microspheres. Normovolemic dogs (n = 10) were subjected to the same protocol except they were not bled. During hypovolemia, naloxone produced significant increases in myocardial, intestinal, hepatic, and adrenal blood flows whereas saline treatment did not. No significant changes in skin, muscle, fat, pancreatic, renal, or brain flows were detected. The increases in blood flow were not associated with significant changes in vascular resistance. Naloxone had no significant effects on any hemodynamic parameter during normovolemia. The beneficial effects of naloxone in hemorrhagic shock include increased blood flow to vital organs due to increased perfusion pressure which is secondary to improved cardiac performance. PMID- 2982169 TI - Temporal relationships between isometric force, phosphorylase, and protein kinase activities in vascular smooth muscle. AB - Vascular smooth muscle contractility is tightly coupled to ATP production by intermediary metabolism. To elucidate mechanisms underlying coordination of metabolism and contractility we studied the time course of isometric force, and the activation of phosphorylase and cAMP-dependent protein kinases during stimulation of bovine coronary arterial strips with KCl. Isometric force reached a maximum after 10 min of exposure to 30 mM KCl (ED90) and was sustained throughout the subsequent 20-min period of contraction. In contrast, activation of phosphorylase was biphasic: enzymic activity reached a maximum (176 +/- 10% of control) after 3 min of contraction and then, though remaining above control, activity declined to a lower level (135 +/- 7% of control). However, no change occurred in the activity ratios for cAMP-dependent protein kinase assessed in either the presence (type II isozyme) or absence (type I isozyme) of 0.5 M NaCl. These data suggest that the activation of phosphorylase during K+-induced contraction is independent of the cAMP system. The biphasic activation of phosphorylase may reflect transient changes in the intracellular concentration of Ca2+ or the activation of a phosphatase(s) during the response. PMID- 2982170 TI - Changing perspectives of the effects of infectious diseases. AB - There are a large number of infectious agents which damage or kill the developing fetus or child. In this report I have summarized the changing status of information available for just six of these diseases. For rubella there has been excellent success with the development of a vaccine for prevention of this disease. Unfortunately the vaccine is only used extensively in a few countries. Herpes simplex has become a particular problem because of the rapid increase in the frequency of genital herpes infections, the spontaneous recurrences of the lesions, and the difficulty in prevention or treating congenital infections. Cytomegalovirus infections are now known to occur in 1% of newborns, and some damage is produced in about 10% of these children. We have no methods for preventing or treating this infection. Varicella at term is also an important risk to the child but this can now be controlled by the use of VZIG. Varicella earlier in pregnancy may be teratogenic. Venezuelan equine encephalitis virus can cause severe brain damage to the fetus, and one of the vaccines for this infection was shown to be teratogenic in monkeys. Lastly, congenital toxoplasmosis has been known for many years to cause fetal damage and death. Further studies of serological detection of infection and treatment are needed. A number of other viral, bacterial, and parasitic diseases must also be considered as part of the total group of these preventable causes of physical and mental damage to the fetus and newborn. PMID- 2982171 TI - Macroglossia as a presentation of the Beckwith-Wiedemann syndrome. AB - Plastic surgeons are not infrequently required to reduce the size of a large tongue in a child. Macroglossia is one of the main presenting features of the Beckwith-Wiedemann syndrome. This comprises a spectrum of disorders that includes a high incidence of malignancy. Patients with this syndrome should be identified and carefully assessed to exclude serious complications. This paper reports a series of 30 patients presenting to the Royal Children's Hospital over an 8-year period. An incidence of malignancy of 10 percent is recorded. PMID- 2982172 TI - Diagnosis and treatment of a delayed sequel to biopsy of a hepatoma. PMID- 2982173 TI - Breast disease in the male: galactographic evaluation. AB - Seven men with unilateral nipple discharge underwent galactography. In two patients the diagnosis was carcinoma, two were benign papillomas, one was a breast abscess, and two were ductal ectasia. Galactography is useful in men and women with nipple discharge, especially when the discharge is bloody and there is no palpable tumor. The precise location of an intraductal lesion through the use of galactography guides the biopsy and makes conservative surgery easier. PMID- 2982175 TI - Immunization with oral poliovirus vaccine in China. PMID- 2982174 TI - Hepatocellular carcinoma with skeletal metastasis. AB - Three patients with symptoms related to metastases from hepatocellular carcinoma are described. The diagnosis of the primary tumor was made at autopsy in two cases and by biopsy in one. The skeletal lesions had a lytic, expansile, and hypervascular appearance. This hypervascularity may lead to bleeding either spontaneously or following biopsy. Hepatocellular carcinoma should be included in the differential diagnosis of osteolytic, expansile, hypervascular metastases, especially when such lesions are encountered in patients with liver cirrhosis. PMID- 2982176 TI - Genetically engineered herpes simplex virus vaccines. PMID- 2982177 TI - [Differential diagnosis of primary bone tumors and tumor-like lesions of the ribs]. AB - Primary bone tumours in ribs are less common than tumour-like lesions. The most common solitary primary bone tumour in ribs is the chondrosarcoma (about 35%). This is followed with about the same frequency (10 to 14%) by Ewing's sarcoma, malignant lymphoma, chondroma and osteo-chondromas. The clinical features and radiological appearances of the more common tumours and tumour-like lesions in the ribs are described, and their differential diagnosis is discussed. PMID- 2982178 TI - [Differential diagnostic value of the hand arteriogram in primary and secondary Raynaud's syndrome]. AB - Arteriograms of the hands were carried out in 348 patients. Functional and organic changes were analysed in an attempt to differentiate primary from secondary Raynaud's syndrome. The value and limitations of this technique are illustrated by a number of examples. PMID- 2982179 TI - Lobus venae azygos and congenital lobar emphysema in an infant. PMID- 2982181 TI - [Diagnosis of gallbladder carcinoma]. PMID- 2982180 TI - [Expectoration of a shell fragment]. PMID- 2982182 TI - [Late perforation after a colonic contrast medium enema]. PMID- 2982183 TI - A complete proatlas, partially assimilated. PMID- 2982184 TI - [Fistula formation after peridural anesthesia. A reliable diagnosis of a rare catheter complication by computed tomography]. PMID- 2982185 TI - [Tuberculosis of the smaller joints]. AB - Eighty-nine patients with tuberculosis of joints in the hands or feet have been examined. There has been a change in the peak incidence from adolescence to adult life. The high number of incorrect diagnoses was remarkable (45% in our own patients). A previous tuberculous infection was found in 40% of the patients. Osteoporosis, or bone atrophy, are important radiological signs for the early diagnosis and follow-up of articular tuberculosis. The later phase is characterised by narrowing of joint spaces. Later still there is destruction of subchondral lamellae and fusion of the carpal and tarsal bones, resulting in changes of shape and position. PMID- 2982186 TI - [Comparative MR and CT research in selected neuroradiological problems]. AB - Magnetic resonance (MR) is a new digital imaging method which is of particular interest in the radiological diagnosis of the central nervous system. It provides a new form for demonstrating the brain and the spinal cord. To evaluate the scope of the method, it was compared with CT. The results in patients with epilepsy of unknown origin, cerebral infarcts, sellar space-occupying lesions and spinal angiomas are described. MR provides information concerning tumours causing epilepsy and also fresh infarcts at an earlier stage than CT. MR has not, so far, provided any new insights in the diagnosis of sellar tumours or spinal angiomas. In these cases, MR can, nevertheless, compete with CT and with conventional radiological methods. Present experience, particularly with problem cases, indicates that MR is becoming an important method of examination in neuroradiology. PMID- 2982188 TI - [Peripheral venous digital subtraction angiography in the follow-up of aortocoronary venous bypass grafts]. AB - 36 patients with 68 aorto-coronary venous bypass grafts (45 of which were angiographically patent) were examined via DSA (DV12V, serial mode, Philips) and angiography. Imaging was triggered via R waves, and effected in RAO 30 degrees, LAO 60 degrees, and AP projection. 63 of the 68 grafts were correctly detected (92.7%). Two anterior interventricular branch grafts (RIVA = ramus interventricularis anterior) were not visualised via DSA (95.6% sensitivity). One graft each to the RIVA, to the right coronary artery (RCA), and to the posterior branch of the lateral cutaneous branches of intercostal arteries (PLA), were falsely found to be patent (87% specificity). Grafts were visualised in full length. In some cases morphological changes of the walls were also visible. PMID- 2982187 TI - [Indications for digital subtraction angiography of the pulmonary vessels]. AB - Different indications of pulmonary DSA are discussed based on experiences in 122 cases. The quality and accuracy of imaging is demonstrated by representative examples. Indications of DSA and conventional pulmonary angiography are principally identical. However, our experience in DSA suggests that this minimal invasive procedure may replace conventional angiography in most cases and abolish the strict indication. Supported by further critical tests the high diagnostic quality of DSA imaging in pulmonary embolism and therapeutical control may substitute radionuclide methods (perfusion-ventilation-scintigraphy) to avoid costs of a double examination. PMID- 2982189 TI - [Transvenous digital subtraction angiography of the thoracic aorta after surgical correction of isthmic stenosis]. AB - 25 patients (normotensive n = 19, hypertensive n = 6) underwent DSA control following corrective surgery of a coarctation of the thoracic aorta to exclude vascular complications. Simultaneously pressure gradients were determined between upper and lower extremities using the doppler-sonography. DSA was diagnostic in all patients. 2 cases showed a patch-dependent dilatation, in 1 case an anastomotic aneurysm could be demonstrated. In 6 patients with hypoplastic aortic arch and 2 patients with a slight reduction of the diameter in the anastomotic region a pathological pressure-gradient was measured. In these patients the ratio of aortic arch rsp. anastomosis/descending aorta was reduced in diameter (less than 0,66) and area (less than 44). Hypertension could not be related to pressure gradients or vascular disorders. PMID- 2982190 TI - [Cavography in the digital subtraction technic]. AB - Experience with 74 patients has shown that digital subtraction angiography is a suitable method for the demonstration of the vena cava. In 28 patients the examination was repeated by the conventional large film angiography procedure, but this added no significant information to help to solve the clinical problem. Electronic contrast enhancement permits lower concentrations of contrast to be used than for conventional techniques. Combined with a more selective use of films, this leads to a reduction in the cost of the examination. PMID- 2982191 TI - [Comparison of Doppler ultrasound, intravenous DSA and conventional film angiography in diagnosing stenosing changes in the area of the carotid bifurcation]. AB - The findings on Doppler ultrasound, intravenous DSA and conventional film angiography were compared in 66 examinations of the carotids with regard to their specificity, sensitivity and accuracy. Comparisons with the literature show that intravenous DSA equals Doppler ultrasound only if good or excellent results of the latter are considered. If poorer DSA results are included, diagnostic precision of this method is 5 to 10% less than that of Doppler ultrasound. Mural ulceration can be diagnosed by means of intravenous DSA in only about 50% of cases, whereas Doppler ultrasound is unable to diagnose this. Improved image quality of intravenous DSA, with better diagnostic precision, could be achieved by using ECG gating. PMID- 2982192 TI - [Efficiency of radiodiagnosis of acoustic neurinomas. Changes in the clarification procedure within the last 10 years]. AB - In a retrospective study we tried to define the diagnostic value of different x ray studies in 43 histologically proved acoustic neuromas. We divided the examinations into such which definitely confirmed a clinical suspicion i.e. formed the last preoperative studies, and into others, which only increased the clinical suspicion. We were especially interested, whether or not Stenvers and transorbital projections of the petrous bones and internal acoustic canals and petrous bone tomographies were essentially confirming clinical diagnosis. Whereas 13 out of 37 Stenvers, 2 out of 5 transorbital projections and 14 out of 18 frontal tomographies increased the suspicion of an existing Neuroma, a true diagnostic confirmation was never possible with these 3 techniques. Vice versa out of 45 computertomographies 33 were confirming the clinical diagnosis and 1 increased the clinical suspicion, with 5 false negatives and 6 nonconclusive studies. CT and Air-CT-Cisternography represent today the definitive diagnostic studies. For economical reasons they should be employed early in the diagnostic course, in those cases with high clinical suspicion or when clinical symptoms suggesting AN persist. PMID- 2982193 TI - [Intracranial venous angiomas]. AB - Until now there have only been 13 reports of clinically diagnosed intracranial venous angiomas, which have been operatively demonstrated and histologically examined. We have seen 2 further cases, both of which received operative treatment. Although this condition is pathologically commoner as arteriovenous angioma, it is rarely clinically diagnosed. The diagnostic value of the computer tomogram is stressed and the findings from 2 young patients are compared with the literature. The angiogram from these angiomas's show considerable differences depending on the method used. This could explain some of difficulties in histological diagnosis. PMID- 2982194 TI - [Bedside chest x-rays and extravascular lung water determination in intensive care patients]. AB - Radiological staging of pulmonary oedema was compared with the determination of extra-vascular lung water by means of a double indicator dilution technique. One hundred and forty-six ward chest radiographs were evaluated and compared with the results of simultaneous measurements of lung water. Seventy-seven cases could be evaluated statistically. Chest x-rays regarded as normal corresponded to extra vascular lung water of 5 to 9 ml./kg. body weight. Interstitial oedema (radiological stage I and II) corresponded to extravascular lung water levels of 8 to 12 ml./kg. Differentiation of stages I and II was not possible. During stage III, extra-vascular lung water was 15 to 21 ml./kg. A comparative analyses of these findings revealed a discrepancy of 34%. The reasons for this are discussed. PMID- 2982195 TI - [Mediastinal CT-staging of bronchial carcinomas]. AB - Forty-three patients with proven bronchogenic carcinoma were evaluated with computed tomography (CT) to search for mediastinal and broncho-pulmonary nodal metastases. Definitive staging was achieved by thoracotomy (34 patients) and mediastinoscopy or transbronchial nodal sampling (9 patients). Three parameters of lymph node appearance on CT were studied to improve the sensitivity and specificity of CT in detecting malignancy in bronchogenic carcinoma: 1) node size, 2) node location and 3) border regularity and definition. 762 clearly defined nodes were studied by CT - 391 metastatic and 371 non metastatic. The most useful CT parameters were node size over 10 mm, and node location, which results in a sensitivity of 80% and a specificity of 88%. A combination of all 3 CT parameters increased sensitivity to 82% and specificity to 90%. These results show clearly, that size is not the only relevant CT-parameter in successful preoperative identification of mediastinal nodal metastases. PMID- 2982196 TI - Percutaneous transhepatic cholecystostomy. AB - Ultrasound-guided percutaneous cholecystostomy was performed in 11 patients. Indications were acute cholecystitis in 7 patients, and obstructive biliary disease in 4 patients. A transhepatic approach was used in all patients. In one patient, bile leakage was noticed during cholecystectomy. No other complications were seen. Three of five patients with a acalculous cholecystitis had a normal post-drainage cholangiogram and the drainage catheter could be removed. A follow up of 5.5, and 26 months respectively, showed no recurrent disease. Three patients underwent surgery and the remaining five patients were drained until death by their underlying disease. Percutaneous cholecystostomy can be a life saving and curative procedure in the critically ill patient, and, in case of acalculous cholecystitis, the only one needed. PMID- 2982197 TI - [Embolization of a pseudoaneurysm of the hepatic artery with hemobilia by direct percutaneous puncture]. AB - Haemobilia is a rare cause of gastrointestinal bleeding that carries a high mortality. It occurs as a complication of percutaneous trans-hepatic biliary drainage in up to 6% cases. The cause is usually a pseudo-aneurysm of the hepatic artery, which can be treated by trans-catheter embolisation. If this fails, direct puncture and embolisation of the aneurysm is a possible alternative to surgical ligation of the hepatic artery. PMID- 2982198 TI - [Improvement in abdominal imaging in sonography by the compression and compound technics]. AB - In a clinical study with 663 patients the influence of compression and compound technique of the visualisation of the abdomen was tested with a real-time-scanner Combison 100, compound scanner Combison 202 and the Octoson-machine. In 66 patients the retroperitoneal space was not demonstrated. Compression with the real-time-transducer improves demonstration in 50% (33/66), compound-technique 20% (19/66). Compound-technique was handicapped in 30% (19/66) by velocity errors and reflection which causes a deterioration in resolution. PMID- 2982199 TI - [Selective coronary angiography via flashing tomosynthesis]. AB - To assess the value and significance of short-term tomosynthesis in judging the pathological condition of coronary vessels, the authors examined 34 patients with coronary heart disease. In comparison with 35 mm cinecoronary angiography, all angiographically determined vascular occlusions and stenoses were identified via short-term tomosynthesis. Comparison of the degrees of stenoses yielded good correlation (r = 0.85). Short-term tomosynthesis offers the advantage of employing smaller amounts of contrast medium to be injected; furthermore, the examination period is shorter, and exposure to radiation is also significantly less. PMID- 2982200 TI - [MR measurements on a perfused renal artery model with pulsatile flow]. AB - MR tomography permits visual demonstration of flow and turbulence. A model was used to compare MR-measured signal intensities and flow profiles as obtained by Doppler anemometry. PMID- 2982201 TI - [Coronary angiographic imaging of tumor vessels in myxoma of the right atrium]. PMID- 2982202 TI - In vitro phosphorylation of rat kidney proximal tubular brush border membranes. AB - The phosphorylation of rat renal brush border membrane protein was analyzed after incubation of cortical slices with 32P-orthophosphate and compared with the phosphorylation by gamma-32P-ATP of isolated brush border vesicles. Phosphate incorporation into brush border membranes isolated from slices was linearly related to the incubation time as well as to the specific activity of orthophosphate present during slice incubation. Incorporation of phosphate into proteins reached an equilibrium after about 60 min, whereas incorporation of phosphate into lipids increased continuously. In brush border membranes isolated from slices incubated with orthophosphate (32P), the addition of cAMP or theophylline produced a dephosphorylation of a 47,000-dalton protein; no increased phosphorylation was observed. In brush border membranes, phosphorylated with gamma-32P-ATP, cAMP and dibutyryl cAMP (dB-cAMP) produced an increase in phosphorylation but no dephosphorylation. Sodium-dependent phosphate transport in brush border membranes was not altered by an incubation of slices with cAMP or dB cAMP. These observations suggest that the phosphorylation machinery of isolated rat renal brush border membranes does not correspond with the mechanisms leading to phosphate incorporation into brush border membrane proteins in the intact cell. PMID- 2982203 TI - Interrelation between oxygen consumption and Na-K-ATPase activity in rat renal proximal tubule suspension. AB - The portion of Na-K-ATPase activity on oxygen consumption was determined in suspensions of rat proximal tubules by monitoring oxygen consumption (QO2) under different metabolic states: in the presence or absence of succinate or lactate (10 mmol/l) and increasing concentrations of ouabain (0.4; 0.8; 1.2 and 1.6 mmol/l). In the metabolic states tested, the ouabain induced decrease of QO2 was identical, which implies a fixed rate between Na-K-ATPase activity and QO2. On the basis of these results, Ki and maximal inhibition rate were determined by Lineweaver-Burk and Eadie-Hofstee plots Ki was 0.67 mmol/l ouabain and the maximal inhibition of QO2 was 77%. This corresponds to an absolute decrease of QO2 of 1,630 mumol O2 . h-1 . g-1 protein. Since this value represents the real portion of Na-K-ATPase on QO2, the activity of Na-K-ATPase can be calculated, yielding an activity of 163 mumol ATP . min-1 . g-1 tubule protein. PMID- 2982205 TI - The oxygen free radical system and myocardial dysfunction. AB - The pathways for the metabolism of molecular oxygen involve one electron-transfer reaction with the subsequent production of reduced-oxygen intermediates. These reduced-oxygen intermediates include the superoxide anion (.O2-), hydrogen peroxide (H2O2), and the hydroxyl radical (.OH), which are highly reactive, short lived species. Normally intracellular enzyme systems that include superoxide dismutase, catalase, and glutathione peroxidase are responsible for "scavenging" these products of oxygen metabolism. However, in many pathological states such as inflammation, ischemia, and reperfusion, there is an increased production of these reduced-oxygen intermediates, which are capable of extensive tissue damage. It is the purpose of this symposium to examine, in depth, the role of oxygen free radical systems as mediators of myocardial dysfunction and expand our knowledge of myocardial ischemia, ischemia-reperfusion injury, and the inflammatory response of the myocardium. PMID- 2982204 TI - Identification of hydrogen peroxide and hydroxyl radicals as mediators of leukocyte-induced myocardial dysfunction. Limitation of infarct size with neutrophil inhibition and depletion. AB - Neutrophil infiltration of the myocardium is an important component of such diverse disease entities as myocarditis, ischemia, and ischemia-reperfusion injury. We have hypothesized that activated neutrophils are capable of disrupting myocardial function via an oxygen free-radical mechanism. Human neutrophils activated with phorbol myristate acetate disrupted calcium transport by canine cardiac sarcoplasmic reticulum, and this process was inhibited by a combination of superoxide dismutase and catalase. In addition, the activated neutrophil system was also inhibited by the combination of cyclooxygenase inhibitors (ibuprofen and indomethacin) and catalase and accelerated by MK-447. These results incriminate both hydrogen peroxide and the hydroxyl radical as mediators of neutrophil-induced myocardial dysfunction. A test of this hypothesis in vivo was performed by neutrophil-depleting dogs with anti-canine leukocyte antisera prior to coronary artery ligation. Following 6 hr of reperfusion, there was a 43% reduction in infarct size compared to non-immune-sera-injected animals. We conclude that oxygen free radicals generated by neutrophils are capable of inducing significant myocardial injury and play an important role in the pathophysiology of ischemia reperfusion injury. PMID- 2982206 TI - Free radicals and myocardial ischemia. The role of xanthine oxidase. AB - Recent studies have established a major role for oxygen-derived free radicals in post ischemic tissue injury to the intestine. During ischemia, there appears to be a calcium-triggered, protease-dependent conversion of the native xanthine dehydrogenase to a superoxide-producing xanthine oxidase. The catabolic degradation of ATP during ischemia provides an oxidizable substrate, hypoxanthine. On reperfusion, molecular oxygen is resupplied and a burst of superoxide production ensues, resulting in extensive tissue damage. The same mechanism appears to occur in myocardial ischemia. Xanthine dehydrogenase rapidly converts to the oxidase during nonperfusion in the rat heart. In the isolated perfused working rat heart model, 40 min of anoxia followed by reoxygenation results in substantial release of creatine kinase. The release of creatine kinase is blocked almost completely by pretreatment of the rats with allopurinol, a specific inhibitor of xanthine oxidase. PMID- 2982207 TI - Oxygen radicals and tissue damage in heart hypertrophy. AB - Cyanide-resistant respiration in heart homogenates supplemented with 1 mM NADH was greater in hypertrophied homogenates (60 days banding) with respect to control homogenates, particularly when the homogenates were incubated in 100% oxygen. The intermyofibrillar mitochondria from hypertrophied hearts produced more superoxide radicals than sub-sarcolemmal mitochondria, and both values were greater than in the unbanded group. H2O2 formation was more evident in the intact mitochondria prepared from hypertrophied hearts than in those of the control hearts. Moreover, the perfusion of isolated hearts in anoxic and reoxygenated conditions caused a greater lipoperoxidative and functional damage at the mitochondrial level in hypertrophied hearts than in the control hearts. These results, correlate with the reduction in mitochondrial function found in the overloaded hearts, suggest an involvement of the reactive species of oxygen in the formation of cardiac damage induced by prolonged aortic banding. PMID- 2982208 TI - Different effects of dietary chenodeoxycholic acid and ursodeoxycholic acid on colonic adenylate cyclase in the rat. AB - The oral administration of dietary chenodeoxycholic acid (1%), but not of ursodeoxycholic acid (1%), to male Sprague Dawley rats results in a significant increase in the colonic adenylate cyclase activity without any influence on the colonic cyclic-AMP phosphodiesterase activity. No effect of chronic bile acid feeding on the response of colonic adenylate cyclase to prostaglandin E2 and vasoactive intestinal peptide is observed. These data emphasize a dependence of the cyclic-AMP adenylate cyclase activation on the chemical structure of the bile acid. This may be of pathophysiologic relevance with respect to the frequently observed diarrhea as a side effect of oral chenodeoxycholic, but not ursodeoxycholic acid therapy for cholesterol gallstone dissolution in man. PMID- 2982209 TI - Amelioration of hydroxyurea-induced suppression of phagocytosis in human granulocytes by free radical scavengers. AB - The exposure of human circulatory white cells in vitro to 0.1-1-10 mol/l hydroxyurea (HU) for 20 h induced a progressive dose-dependent suppression of the phagocytic activity of granulocytes. The suppressing effect of 20 h exposure to 1 mol/l HU was used to examine the protection afforded by free radical scavengers against HU-induced cytotoxicity. It has been found that, in the suitable concentration of the protecting agent, a substantial protective effect of sodium benzoate, acetylosalicylic acid, alpha-tocopherol, ascorbic acid, catalase, peroxidase or superoxide dismutase can be achieved. PMID- 2982210 TI - Use of restriction fragment length polymorphisms to determine the clonal origin of human tumors. AB - A novel strategy to determine the clonal origin of human tumors has been devised. The strategy involves the use of a cloned polymorphic X-chromosomal gene and two restriction endonucleases. The first endonuclease distinguishes the paternal and maternal copies of the gene through a DNA polymorphism of restriction fragment length. The second endonuclease distinguishes active from inactive copies of this gene through changes in DNA methylation. As illustrations of this strategy, three human cancers were each shown to be monoclonal. The analysis described should have a wide variety of clinical and experimental applications. PMID- 2982211 TI - Oxidative autoactivation of latent collagenase by human neutrophils. AB - The pathological destruction of collagen plays a key role in the development of inflammatory disease states affecting every organ system in the human body. Neutrophils localized at inflammatory sites can potentially degrade collagen by releasing a metalloenzyme, collagenase, which is stored in a latent inactive form. Triggered human neutrophils were shown to release and simultaneously activate their latent collagenase. The activation of the latent enzyme was coupled to an oxidative process that required the generation of a highly reactive oxygen metabolite, hypochlorous acid. Oxidative regulation of latent collagenase activity may be important in the pathogenesis of connective tissue damage in vivo. PMID- 2982212 TI - Induction of DNA synthesis in cultured rat hepatocytes through stimulation of alpha 1 adrenoreceptor by norepinephrine. AB - Addition of norepinephrine to primary cultures of adult rat hepatocytes stimulates the incorporation of [3H]thymidine in a dose-dependent manner. This effect has been observed in serum-free medium containing epidermal growth factor and insulin. Stimulation of DNA synthesis by norepinephrine was strongly antagonized by the alpha 1-adrenergic antagonist prazosin but not by an alpha 2 antagonist or by a beta-adrenergic blocker. The beta agonist isoproterenol did not stimulate significant DNA synthesis. These results indicate that catecholamines interact with the alpha 1 adrenoreceptor to stimulate DNA synthesis in hepatocytes. Since alpha 1 receptors are present in most cells, this receptor may be important in cell growth regulation. PMID- 2982213 TI - Protection against lethal hyperoxia by tracheal insufflation of erythrocytes: role of red cell glutathione. AB - Intact erythrocytes placed into the tracheobronchial tree of hyperoxic rats dramatically improved their chances for survival. Over 70 percent of the animals so treated survived more than 12 days during continuous exposure to 95 percent oxygen, whereas all of the control animals died within 96 hours. Lungs from erythrocyte-protected rats showed almost none of the morphologic damage suffered by untreated animals. Erythrocytes containing cyanomethemoglobin were as beneficial as normal erythrocytes, but cells in which glutathione was partially blocked were significantly less protective. Analogous results were obtained in vitro: 51Cr-labeled target cells released 70 to 90 percent of their label when exposed briefly to hydrogen peroxide or to toxic oxygen species generated by phorbol ester-stimulated neutrophils. Addition of intact erythrocytes decreased release by approximately 75 percent, but significantly less than this if red blood cell glutathione was partially blocked. These results suggest that insufflated erythrocytes, through their recyclable glutathione, protect rats from toxic oxygen species engendered by hyperoxia. PMID- 2982214 TI - Distinct hepatic receptors for low density lipoprotein and apolipoprotein E in humans. AB - Since the liver is a central organ for lipid and lipoprotein synthesis and catabolism, hepatic receptors for specific apolipoproteins on plasma lipoproteins would be expected to modulate lipid and lipoprotein metabolism. The role of hepatic receptors for low density lipoproteins and apolipoprotein E-containing lipoproteins was evaluated in patients with complementary disorders in lipoprotein metabolism: abetalipoproteinemia and homozygous familial hypercholesterolemia. In addition, hepatic membranes from a patient with familial hypercholesterolemia were studied and compared before and after portacaval shunt surgery. The results establish that the human liver has receptors for apolipoproteins B and E. Furthermore, in the human, hepatic receptors for low density lipoproteins and apolipoprotein E are genetically distinct and can undergo independent control. PMID- 2982215 TI - "Anxiety peptide" found in brain. PMID- 2982216 TI - Limitations of computed tomography following excisional biopsy of soft tissue sarcomas. AB - Twenty-one patients were evaluated by computed tomography (CT) following complete or incomplete excisional biopsy of soft tissue sarcomas. Since the surgical margins were inadequate, additional treatment was required, and CT was intended to identify and delineate any residual tumor. Thirteen patients had no palpable mass in the operative area. In these, eleven CTs showed no tumor, but microscopic tumor was found in seven. The other two CTs showed masses, but both proved to be hematomas. Eight patients had palpable masses and seven proved to be residual tumor. Of these, two CTs failed to show the residual tumor. Five CTs correctly identified residual tumor, but two of them failed to show the entire tumor extent. Computed tomography did not correctly predict the presence or absence of microscopic residual tumor when there was no palpable mass, and was at least partly inaccurate in delineating residual tumor in four of eight patients with palpable masses. PMID- 2982217 TI - [Study of space-occupying lesions in the liver using technetium-99m tin colloid and indium-113m chloride]. AB - Liver scanning with radiocolloids is an important method to determine the presence, the position and the size of space-occupying lesions in the liver. Unfortunately, this information is nonspecific and it is not possible to distinguish between tumours, abscesses or cysts. Thirty-six patients in whom a definite diagnosis of hepatoma, amoebic liver abscess or echinococcus cyst had been made were examined with technetium-99m tin colloid and indium-113m chloride. The amoebic liver abscesses were avascular, showed a hyperaemic area surrounding the abscess and appeared smaller on the indium than on the technetium scan. The hepatomas showed greater vascularity and absence of the hyperaemic area. Cysts were avascular, did not show a hyperaemic rim and the size was equal on both scans. The experience of the observers had an influence on the accuracy of interpretation of the scans; experienced observers made a correct diagnosis in 73% of cases. It is suggested that simultaneous 99mTc tin colloid and 113mIn chloride scans provide additional specificity in the differential diagnosis between hepatoma, amoebic liver abscess and echinococcus cysts. PMID- 2982218 TI - The management of patients with carcinomas in fibroadenomatous tumors of the breast. AB - This study focuses upon 38 patients found to have unsuspected carcinomas or lobular neoplasia in fibroadenomatous tumors. Combining the pathologic and clinical features of these patients with those of 56 comparable instances reported upon in the literature, it becomes apparent that the biologic and clinical behavior of lobular neoplasia, intraductal carcinomas and invasive carcinomas is essentially the same regardless of whether the lesions arise in or outside a fibroadenoma. In addition, they are often a part of a multifocal malignant condition in the ipsilateral or contralateral breast. These patients, therefore, should be treated as if the carcinomas or lobular neoplasia arose in the usual setting. PMID- 2982219 TI - [Human T-lymphotropic retrovirus and AIDS. HTLV-III is probably the cause of acquired immunodeficiency syndrome]. PMID- 2982220 TI - [Pain and endorphins]. AB - Various stimuli may evoke the symptom pain. These stimuli activate sensory receptors and this information is transmitted to the spinal cord and a number of cerebral centres. Various endogenous analgesic systems are present in the body, which can modulate and control the consequences of pain stimuli to some extent. Endogenous morphine-like substances (endorphins) are an essential factor in some of these systems. Endorphins may be released in the body during stress or by acupuncture and other pain-relieving procedures (e.g. the twitch in horses). The significance of endorphins is not confined to pain, but they are also involved in reward systems of the brain may therefore play a role in eating, sexual, social and addictive behaviour among others. PMID- 2982221 TI - [Radiation dose to the lung following endolymphatic radionuclide therapy of melanoma of the lower limbs]. AB - The authors present 230 patients submitted to postoperative intralymphatic radiotherapy (ELRT) with radioactive Lipiodol UF because of a malignant melanoma of the inferior extremity. The incidence and volume of a possible invasion of the contrast medium into the lung and the resulting radiation exposure is indicated. Even if very low quantities of the contrast medium (3.5 ml and 7.0 ml, respectively) are used, only 22% of the cases show no pulmonary activity at all. Those patients who underwent bipedal ELRT showed more frequently higher radiation exposure. A correlation between lymph node weight and radiation exposure of the lung could not be demonstrated. PMID- 2982222 TI - Does Cohn-fractionated Rh immune globulin transmit viral hepatitis? AB - In light of recently raised doubts about the safety of Cohn fraction II globulins, a prospective study on the risk of transmission of viral hepatitis with a Cohn-fractionated Rh immune globulin (Rhesonativ, KabiVitrum AB, Stockholm, Sweden) was performed in 47 newly delivered mothers. The women were followed regularly for 6 months after the injection of the Rh immune globulin for biochemical, serological, and clinical signs of viral hepatitis. No clinical signs of acute hepatitis were noted during the study, nor were HBsAg or anti-HBc found in any patient. A slight and transient rise in alanine aminotransferase (ALT) levels was seen in three women, but these never reached 2.5 times the upper normal limit as is the currently used lower limit for a diagnosis of non-A, non-B hepatitis. One woman had positive tests for anti-HBs at 5 and 5.5 months, respectively, after the injection, but serum samples taken before and after this period were all anti-HBs negative. Nonspecific reactions in the method used probably explained this finding. This prospective study supports the contention that Rhesonativ, a Cohn-fractionated Rh immune globulin, does not transmit viral hepatitis. PMID- 2982223 TI - Changes in human cultured cells exposed to a perfluorocarbon emulsion. AB - A perfluorocarbon emulsion (Fluosol-DA, 20%) produced persistent cytotoxic changes and growth inhibition in fibroblast-like human cells. After 18 hours of exposure to culture medium containing 4 percent of this perfluorochemical emulsion, normal embryonic lung fibroblasts (IMR 90 cells) and their SV40 virus transformed counterparts (AG 2804 cells) ceased proliferation and showed degenerative changes, even if Fluosol was washed off the cell monolayer and replaced with normal medium. The morphological manifestations of Fluosol cytotoxicity included cytoplasmic vacuolation of varying but frequently marked degree. These findings raised concerns about the use of perfluorochemicals in patients until safe dose limits can be established. PMID- 2982224 TI - Peripheral blood reconstitution. PMID- 2982225 TI - Antenatal Rh immunoglobulin--help or hindrance? PMID- 2982226 TI - [Constipation during pregnancy. Treatment with Dumovital fiber tablets]. PMID- 2982227 TI - Primary aldosteronism treated by trilostane (3 beta-hydroxysteroid dehydrogenase inhibitor). AB - The practicability and tolerability of trilostane, a competitive inhibitor of 3 beta-hydroxysteroid-delta 5-dehydrogenase, for the therapy of primary aldosteronism was assessed in 1 patient with aldosterone-producing adenoma (APA) and 3 subjects with idiopathic adrenal hyperplasia (IHA). Trilostane afforded reduction of plasma levels of aldosterone, progesterone, deoxycorticosterone, 17 OH progesterone, cortisol, delta 4-androstenedione, and urinary excretion of 17 hydroxycorticosteroid. Conversely, circulating levels of dehydroepiandrosterone, dehydroepiandrosterone sulfate, and urinary excretion of 17-ketosteroids were increased following this drug therapy. Suppression of mineralo- or glucocorticoid biosynthesis was accompanied by an increase in plasma renin activity. One patient with APA or 3 subjects with IHA showed slight or remarkable improvement of hypertension and hypokalemia. Based on these findings, efficacy and tolerability of trilostane appear to aid in the treatment of IHA. PMID- 2982229 TI - [Electron microscopy of bovine leukosis viruses (BLV) in a fetal lamb spleen cell line]. AB - The ultra-thin section method, the method of negative staining, and immunoelectron microscopy were used for detecting BLV and for determining its morphological characteristics in the FLS continual cell line used as a virus antigen producer for the ELISA test. Particles of C type, about 110 nm in size, having a structure corresponding to BLV, were detected in the FLS cells on the ultra-thin sections. The viruses were located extracellularly, in cytoplasmic vacuoles, and in different stages of maturation by budding from cell plasma membrane. BLV presence was also demonstrated by immunoelectron microscopy. PMID- 2982228 TI - [Role of reflex from the vertebral artery on the blood supply of the internal ear]. PMID- 2982230 TI - Islet cell carcinomas in dogs. AB - Seven cases of dog islet cell carcinomas were studied by conventional and immunohistochemical light- and electron-microscopy. Antisera to insulin, pancreatic polypeptide, somatostatin and glucagon were used. In 6 tumours several hormones were demonstrated. Glucagon never occurred. Insulin was the only hormone present in every tumour, thus it seems to be a good marker for these neoplasms. Liver metastases contained less immunoreactive cells than primary tumours and cell types found in primary carcinomas were sometimes not present in liver metastases. In two cases a degenerative neuropathy occurred. PMID- 2982231 TI - Characterization and physical map of choleraphage phi 149 DNA. AB - Choleraphage phi 149 DNA is a linear double-stranded molecule 69 X 10(6) Da or 104 kilobase pairs (kbp). From restriction enzyme analysis, it has been concluded that the DNA is circularly permuted. There are at least three S1 nuclease sensitive sites along the length of the molecule. These sites represent single strand interruptions repairable by T4 DNA ligase. A physical map of the DNA has been constructed using the restriction endonucleases BamH1 and BglII. PMID- 2982233 TI - Host range of mink cell focus-inducing viruses. AB - The species host range of the recombinant, mink cell focus-inducing (MCF) class of murine retroviruses was determined in vitro and compared to the host range properties of xenotropic and amphotropic murine viruses. In contrast to xenotropic and amphotropic viruses, MCF viruses were restricted in the number of mammalian species they would infect. Cell lines from mouse, rat, mink, ferret, and cat were susceptible to MCF infection and certain virus isolates could infect rabbit cells, but cells from Chinese hamster, buffalo, bat, dog, monkey, and human were resistant to infection by most MCF viruses. The resistance of some of the latter cells was abrogated by phenotypic mixing with xenotropic virus, which demonstrated that MCF species host range was mediated by virus envelope-cell surface interaction. The host range uniformity of the various MCF isolates and the unique species distribution of sensitivity are consistent with the conclusion from other evidence that the MCF viruses comprise a class distinct from xenotropic and amphotropic viruses. PMID- 2982232 TI - The human cellular abl gene product in the chronic myelogenous leukemia cell line K562 has an associated tyrosine protein kinase activity. AB - Three antisera against the mouse v-abl gene product were used to identify two potential human c-abl gene products in the chronic myelogenous leukemia cell line K562. Two antipeptide sera were generated in rabbits using the predicted amino acid sequence of the mouse v-abl gene product. One antiserum was made against a polypeptide overlapping the in vivo tyrosine phosphorylation site of murine P120gag-abl and what is believed to be a homologous tyrosine phosphorylation site of the predicted normal human c-abl gene product (v-abl 263-280). The second antipeptide serum, abl 389-403, was generated against a predicted hydrophilic peptide of the v-abl gene product. Immunoprecipitation from K562 cells metabolically labeled with [32P]orthophosphate by a mouse tumor regressor and abl 389-403 antipeptide sera detected two proteins of 190,000 and 240,000 Da. Both proteins were labeled primarily at serine and, to a much lesser extent, at tyrosine residues. Immune complex kinase assays using conditions that allow the tyrosine phosphorylation of P120gag-abl showed that in vitro phosphorylation of P190 and P240 occurs primarily at tyrosine residues. The detection of these enzymatically active human c-abl gene products is a rare observation which may be in part attributed to the c-abl gene translocation from chromosomes 9 to 22 occurring in the vast majority of chronic myelogenous leukemia patients. PMID- 2982234 TI - Base mutations in the terminal noncoding regions of the genome of vesicular stomatitis virus isolated from persistent infections of L cells. AB - The 3'-terminal regions of the genomes of vesicular stomatitis virus obtained from two long-term, independently initiated persistent infections of L cells were found to contain several sequence mutations. In contrast to the hypermutability displayed in the 5'-terminal regions of the genomes of viruses obtained from persistent infections of baby hamster kidney (BHK) cells (P. J. O'Hara, F. M. Horodyski, S. T. Nichol, and J. J. Holland, J. Virol. 49, 793-798, 1984), no 5' mutations were detected in viruses from L-cell carrier lines. The absence of detectable defective interfering (DI) particles in the L-cell carrier cultures may account for this difference. Plus-strand leader RNA made by the viruses from persistently infected L cells failed to accumulate from 5 to 8 hr postinfection unlike the accumulation noted for the leader RNA generated by wild-type VSV. Minus-strand leader RNA, on the other hand, accumulated at a similar or increased rate compared to wild type. The relationship of these observations to the processes of host shutoff, viral transcription, and replication are discussed. PMID- 2982235 TI - Transformation of erythroid cells by Rous sarcoma virus (RSV). AB - RSV transforms several nonhematopoietic cell types and as reported here also has the capacity to transform hematopoietic cells of the erythroid lineage. In vitro, the three RSV isolates tested induced erythroblast-like colonies in infected bone marrow cells that were distinguishable by size and cell arrangement from those induced by avian erythroblastosis virus (AEV). Also in contrast to AEV transformed erythroblast cultures, isolated cell colonies induced by RSV required complex growth conditions in liquid medium similar to the in vitro conditions necessary for erythroblasts transformed by the acute leukemia virus E26. Temperature-shift experiments using temperature-sensitive (ts) NY68 RSV revealed that when grown at the nonpermissive temperature (42 degrees), mutant-infected cells became benzidine positive and partially differentiated into erythrocytes. Wild-type (wt) RSV-transformed cells did not undergo similar changes. However, both wt RSV-, and to a greater extent, ts RSV-transformed cultures at the permissive temperature (37 degrees) did contain populations of spontaneously differentiating erythroid cells signifying that the transforming activity of the virus did not fully arrest erythroid maturation. In addition, the RSV-transformed cells did express tyrosine kinase activity. When injected intravenously into birds, RSV induced an erythroblastosis-like disease similar to AEV but also caused fibrosarcomas and leg paralysis. These results show that RSV can alter the pattern of erythroid differentiation in a manner similar to, but distinct from, AEV and indicate that the tyrosine-specific pp60src kinase is involved in erythroid cell transformation. Since the src and erb B proteins share a significant amino acid homology, these data suggest that both may also share a common functional homology. PMID- 2982236 TI - Purification and chemical and immunological characterization of avian reticuloendotheliosis virus gag-gene-encoded structural proteins. AB - Five gag-gene-encoded structural proteins, designated p12, pp18, pp20, p30, and p10 were purified from replication-competent avian reticuloendotheliosis associated virus (REV-A) by high-performance liquid chromatography complemented with chloroform-methanol extraction and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Based on amino acid composition and NH2- and COOH-terminal sequence analysis p12, pp18, p30, and p10 are distinct from one another, whereas pp20 is likely identical to pp18 in primary structure. The p12 was resistant to Edman degradation and was found to be myristylated at the NH2-terminal amino group. Sequence comparisons among the retrovirus family show that pp18/pp20 and p10 are, respectively, homologs of phospho-proteins and nucleic acid-binding proteins. A comparison of terminal sequences with the nucleotide sequence of spleen necrosis virus (SNV) revealed that the gag genes of SNV and REV-A are highly conserved; together with the identification of REV-A gag-precursor polyprotein, Pr60gag in immunoprecipitates of radiolabeled cell lysates, this comparison also led to the establishment of the organization of Pr60gag, viz., NH2-p12-pp18-p30-p10-OH. Sequence comparisons show that REV-A/SNV is related to mammalian type C viruses: the pp18-p30 region is most homologous to the macaque/colobus group and least to simian sarcoma virus (SSV), whereas both the 5'- and 3'-gag regions (i.e., p12 and p10) are clostest to SSV. Immunological studies using monospecific antisera and Western-blot analysis showed that antigenic determinants of REV-A p30 are conserved in most of mammalian type C and type D viruses, but those of REV-A p12 are shared only with simian sarcoma associated virus (SSAV) and endogenous viruses of macaques. PMID- 2982237 TI - New bacteriophage-like particles in Corynebacterium glutamicum. AB - Three new phage-like particles (CG1, CG2, and CGK1) were isolated from Corynebacterium glutamicum CBII. Particles CG1 and CG2 are DNA phages with long, noncontractile tails, CGK1 is a killer particle according to electron microscopy. A heat-stable low-molecular-weight bacteriocidal substance affecting various coryneform bacteria was observed to be joined to the killer particle CGK1. PMID- 2982238 TI - Activation of the cholinergic system and growth hormone release in the dog: functional interactions with other neurotransmitters. AB - In unanaesthetized dogs iv administration of the cholinesterase inhibitor eserine (0.5 mg) induced a clear-cut rise in plasma canine growth hormone (cGH) levels. Diphenhydramine and meclastine, two antagonists of histamine (H) H1 receptors completely suppressed the GH-releasing effect of eserine, while cimetidine, an H2 receptor antagonist, only blunted and delayed it. Two long-lasting serotonin (5 HT) receptor antagonists, metergoline and pizotifen, partially or completely suppressed, respectively, GH release evoked by eserine, whereas fenfluramine, a releaser of neuronal stores of 5-HT and hence a functional activator of 5-HT neurotransmission, was ineffective in this context. Pimozide, a long-acting dopamine receptor antagonist, abolished the effect of eserine, whereas domperidone, which has the same pharmacological properties but does not cross the blood brain barrier, failed to do so. Finally, phentolamine, an antagonist of alpha-adrenoceptors, and propranolol, a beta-adrenergic antagonist, were completely ineffective in preventing the rise in plasma cGH levels induced by eserine, as was naloxone, an antagonist of opiate receptors. All these data demonstrate that, although cholinergic mechanisms are involved in the mechanism(s) underlying cGH release, the final common pathway for GH secretion is not cholinergic. Preservation of dopaminergic and H1 neurotransmission, probably within the blood barrier, is needed to allow the neuroendocrine transduction of cholinergic inputs, whereas the role of 5-HT neurotransmission remains uncertain. PMID- 2982239 TI - Prolonged ACTH infusion suppresses aldosterone secretion in spite of high renin activity. AB - Five normal young males on a low sodium diet received iv ACTH (1-24) infusions (10 IU/24 h) for 100 h in addition to diuretics. The aim of the study was to find out whether the biphasic effect of ACTH on aldosterone (initial stimulation followed by 'escape') could be prevented by keeping plasma renin activity (PRA) at a fairly constant high level. PRA was around 20 ng/kg/min before and towards the end of the ACTH infusion. Plasma aldosterone and aldosterone excretion rates were, nevertheless, only transiently stimulated, but the first was relatively more suppressed than the latter at the end of the ACTH infusion. Plasma 18-OH corticosterone followed the same pattern. Even on the last infusion day, aldosterone and 18-OH-corticosterone levels were still higher than in normal ambulatory sodium-replete men. The fasciculata steroids cortisol, 11 deoxycorticosterone and corticosterone were continuously stimulated by ACTH. It is concluded that the biphasic response of zona glomerulosa steroids to ACTH is basically independent of renin and angiotensin II. However, the marked suppression of aldosterone secretion observed in sodium-replete individuals during prolonged ACTH treatment was not seen in this study. Angiotensin II or a different factor associated with sodium depletion may, therefore, partly protect the zona glomerulosa from adverse effects of ACTH observed in the sodium-replete state. PMID- 2982240 TI - Trophic effects of potassium loading on the rat zona glomerulosa: permissive role of ACTH and angiotensin II. AB - The trophic effects of chronic potassium loading on the rat zona glomerulosa were investigated by morphometric and radioimmunological methods. Potassium loading exerted a potent adrenoglomerulotrophic effect in saline treated control rats, but it was not able to reverse the captopril- and dexamethasone-induced atrophy of the zona glomerulosa. However, if the captopril/dexamethasone administered rats were given maintenance doses of angiotensin II and ACTH, potassium loading was found to exert a strong trophic action. The hypothesis is advanced that potassium loading requires the integrity of both the renin-angiotensin system and the hypothalamo-hypophyseal axis to exert its powerful direct stimulating effect on the growth and steroidogenic capacity of the rat zona glomerulosa. PMID- 2982241 TI - Different alterations of nuclear triiodothyronine receptor capacity in liver and kidney induced by starvation and triiodothyronine administration. AB - Many studies have shown alterations in the number of nuclear triiodothyronine receptor (NT3R) under pathophysiologic situations. Most of these studies were performed on the rat liver and it is not known whether NT3R in different tissues exhibits an alteration similar to that in the liver. We compared the change of nuclear receptor capacity for T3 in the liver and kidney during starvation and after T3 injection. Fasting for 72 h decreased maximal binding capacity (Cmax) in the rat liver receptor to 67% of the control, while it did not significantly change Cmax in the kidney. These changes in Cmax were parallel to those of nuclear protein concentrations in both tissues. Daily sc injection of T3 (20 micrograms/100 g body weight) for 3 days also caused the different alteration of Cmax in the liver and kidney. After T3, hepatic NT3R increased to 182% of the control, but renal NT3R increased only to 136%. Association constants were the same in all groups. These results show that changes of NT3R capacity under some conditions vary in different tissues. PMID- 2982242 TI - The effects of parathyroid hormone on osteoblast-like cells from embryonic chick calvaria. AB - A bone cell fraction was isolated from 16 day old embryonic chick calvaria using a sequential enzymatic digestion procedure. The fraction contained cells, of an osteoblast-like character, which responded to parathyroid hormone (PTH) and prostaglandin E2, but not to calcitonin, in terms of increased production of cyclic AMP. Primary cultures of cells maintained their responsiveness to PTH for at least 2 weeks after reaching confluence. Production of alkaline phosphatase by the bone cells was inhibited when 1,25(OH)2 vitamin D3 was added to cultures at concentrations of 10(-8)M or greater. When cells were cultured in the presence of PTH a biphasic effect was observed; alkaline phosphatase levels were stimulated at low concentrations of this hormone but were decreased at higher concentrations. The latter finding appears consistent with observations that PTH can in vivo exert either anabolic or catabolic effects on bone, depending upon the circulating level of hormone present. PMID- 2982243 TI - Effect of naloxone on oxytocin-induced cortisol decrease in normal men. AB - iv administration of oxytocin decreases plasma ACTH-cortisol levels in normal men. In contrast, naloxone, a specific opioid antagonist, stimulates cortisol release, suggesting that opioid peptides exert an inhibitory control on ACTH cortisol secretion. The present study was carried out in an attempt to determine whether an opioid pathway mediates oxytocin action; therefore, we evaluated the effect of naloxone on the decrease of cortisol induced by oxytocin. Six normal men were treated iv with oxytocin (2 IU as a bolus), naloxone (4 mg as a bolus plus 10 mg infused for 2 h) or a combination of the 2 drugs. Plasma cortisol levels were determined in samples taken before and 2 h after drug treatment. As expected, administration of oxytocin significantly decreased cortisol secretion, while naloxone had a stimulatory effect on plasma cortisol levels. When oxytocin injection was followed by administration of naloxone, cortisol levels remained unchanged; thus, naloxone abolished a cortisol decrement in response to oxytocin. These findings show that in man oxytocin requires an active opioid system in order to produce its inhibitory action on ACTH-cortisol secretion, suggesting that this effect of oxytocin could be mediated by an opioid pathway. PMID- 2982244 TI - Further studies on the nature of neural intranuclear inclusions (Roncoroni bodies) (with 2 color plates). AB - It is generally accepted that chromosomes are not visible in interphase except in certain diptera, amphibia and rare unicellular organisms where they are represented by polytene, giant chromosomes. Such structures are exemplified by cells in which growth is evident by increase in size while the polytene structures increase in size rather than in number. These giant chromosomes are active metabolically, synthesizing mRNA and causing increasing activity in the corresponding cells. Experimental procedures leading to increased activity in neurones were accompanied by the appearance of nuclear inclusions which behaved very much like the polytene chromosomes metabolically and, as a result, an attempt was made to determine whether they had similar chemical behavior. This proved to be demonstrable and so it is concluded that the Roncoroni body represents a metabolically active interphase chromosomal segment and may be used as a sign of such activity. PMID- 2982246 TI - Vitamin D3 stimulates calcium-45 uptake by isolated mouse islets in vitro. AB - Islets isolated from ob/ob mice which had been fed a vitamin D-deficient diet released significantly less insulin in response to glucose than did vitamin D replete islets but showed normal net 45Ca2+ uptake. To determine whether vitamin D3 has a direct effect on the pancreatic B cell, islets from ob/ob mice on a normal diet were exposed to vitamin D3 in vitro for 1 week or only 3 h, and then glucose-stimulated 45Ca2+ uptake and insulin release were measured. Exposure to 1 nM or 1 microM vitamin D3 for 1 week stimulated 45Ca2+ uptake in the presence of 3 mM, but not 20 mM glucose, and did not affect insulin release. Exposure to vitamin D3 for 3 h did not significantly increase net 45Ca2+ uptake although there was a tendency to such an effect (P = 0.10). In conclusion, vitamin D deficiency in vivo suppressed subsequent glucose-stimulated insulin release in vitro and this effect may be due to a direct effect of the sterol (or one of its metabolites) on calcium handling by the B cell. PMID- 2982245 TI - The effect of adrenaline infusion on the regulation of glycogenolysis in human muscle during isometric contraction. AB - The regulation of glycogenolysis in human muscle during isometric contraction without and with adrenaline infusion has been investigated. The content of cAMP in muscle increased three-fold during the infusion. Total glycogen phosphorylase and synthetase activities were unchanged during contraction without and with adrenaline infusion. The fraction of phosphorylase in the a form was in resting muscle 26% and at the end of contraction 24%. During adrenaline infusion phosphorylase a increased to 80%. Contraction during continued infusion resulted in a decrease of phosphorylase a to 42%, despite persistently increased cAMP content in muscle. The activity of synthetase I decreased to about half of the initial value during adrenaline infusion and contraction both without and with the infusion. The rate of glycogenolysis in muscle during contraction was not significantly changed by the infusion. Phosphocreatine (PCr) decreased during the contraction and the decrease was similar without and with adrenaline infusion. The amount of inorganic phosphate (Pi) accumulated in muscle during contraction was lower when adrenaline was given due to a greater accumulation of hexose monophosphates. It is concluded that the rate of glycogenolysis in muscle during contraction without and with adrenaline infusion is a function both of phosphorylase in the form a form and the availability of Pi at the active site of the enzyme. PMID- 2982248 TI - Atrophy of the left hepatic lobe caused by a cholangiocarcinoma. PMID- 2982247 TI - Dependency of renal potassium excretion on Na,K-ATPase transport rate. AB - Potassium secretion may depend on the transport rate of Na, K-ATPase in basolateral cell membranes of distal tubular cells. To examine this hypothesis experiments were performed in anaesthetized dogs during inhibition of proximal potassium reabsorption by acetazolamide or mannitol (fractional potassium excretion 1.2 - 1.4) or additional stimulation of potassium secretion by ethacrynic acid (fractional potassium excretion 2.1). Ouabain in a dose which inhibits 70-80% of the Na, K-ATPase activity reduced fractional potassium excretion to 0.8 - 0.9 by an effect on distal tubular secretion since potassium transport in the proximal tubules was not affected. Ouabain-sensitive potassium excretion varied in proportion to ouabain-sensitive sodium reabsorption during variation in glomerular filtration rate, even at urinary sodium concentrations exceeding 80 mmol X 1(-1). In experiments without ouabain, saline infusion raised potassium excretion and sodium reabsorption until maximal Na,K-ATPase transport rate was reached, as judged from heat production measurements, but not during further increments in urine flow. After inhibition of Na,K-ATPase activity by hypokalaemia, potassium excretion and cortical heat production remained constant over a wide range of urine flow and sodium excretion. We conclude that potassium secretion is dependent on intact Na,K-ATPase activity and is stimulated by sodium delivery to the distal nephron until maximal transport rate of the enzyme is reached. PMID- 2982249 TI - Demonstration of an angiographically hypovascular insulinoma with intraarterial dynamic CT. PMID- 2982250 TI - Proton imaging and phosphorus spectroscopy in a malignant glioma. PMID- 2982251 TI - Radiologic characteristics of primitive neuroectodermal tumors in children. AB - Radiographic findings are reviewed in 31 children with primitive neural ectodermal tumors seen at the Hospital for Sick Children from 1962 through 1983. Seventeen children were initially evaluated with computed tomography (CT). Ten of these had both CT studies and angiography. The tumors were large, irregular, typically iso- to hyperdense, and showed dense, heterogeneous contrast enhancement. Cysts were present in 65% and calcifications in 71% of cases. The angiographic findings were nonspecific, ranging from avascular to markedly vascular. Although these tumors were usually found in the cerebral hemisphere, particularly the frontal lobes, two cases are included of tumors arising elsewhere: one in the pineal and one in the suprasellar region. Epidemiologic data are reviewed. PMID- 2982252 TI - Influence of calcium-channel blockers on platelet function and arachidonic acid metabolism. AB - Available data indicate that platelet function and arachidonic acid metabolism are important factors in hemostasis and regulation of vascular tone. Plasma membrane and intracellular mobilization of calcium ions are intimately related to platelet activation and release of platelet contents. Release of arachidonic acid from membrane phospholipids as well as subsequent synthesis and release of vasoconstrictor thromboxane A2 are also regulated by movement of calcium ions. Adenosine 3':5'-cyclic phosphate in turn controls levels of free calcium ions in platelets and regulates calcium-dependent reactions. Slow-channel calcium blockers, such as verapamil, diltiazem and nifedipine, inhibit platelet activation in vitro, and decrease platelet adhesion intravascularly. These agents have also been shown to decrease platelet nucleotide release and thromboxane A2 generation. Some preliminary data suggest that calcium blockers also increase generation of vasodilator and platelet antiaggregant prostacyclin, which could contribute to decrease in platelet function. These effects of calcium blockers on platelet function and arachidonic acid metabolism could contribute in part to their efficacy in patients with ischemic heart disease. PMID- 2982253 TI - A simple, inexpensive method of determining total body water using a tracer dose of D2O and infrared absorption of biological fluids. AB - An improved infrared spectrophotometric method using tracer doses of D2O for determination of total body water (TBW) is described. Evaluation of sample preparation procedures showed that only vacuum sublimation yielded acceptable recoveries of D2O standards in the range of 0.01-0.30 mg/ml in urine and plasma (101 +/- 2.5 and 99.6 +/- 2.6%, mean +/- SD, respectively). Oral administration of a 10 g dose of D2O was shown to equilibrate within 2 hr in the saliva and plasma of 10 healthy men and women, including obese (30% body fat) subjects. Calculated TBW was 39.1 +/- 6.4 L which represented 74 +/- 1.6% of the fat free mass determined by hydrodensitometry. The precision of the described infrared method was 2.5%. Based upon the observed sensitivity of this method, it would be possible to administer smaller oral D2O doses, 5-6 g, and obtain reliable TBW values. The practical advantages of this method are low cost and a simple analysis that permits repeated TBW measurements over brief periods without an undue buildup of background deuterium levels in the body. PMID- 2982254 TI - Giant cell fibroblastoma: a report of three cases. AB - Three cases of giant cell fibroblastoma are presented. All three patients were boys younger than four years of age. The neoplasms involved the subcutaneous tissue of the back, perineum, and shoulder, respectively. The tumors were characterized histologically by a mixture of spindle cells and multinucleated giant cells in a myxoid or collagenous background and by the presence of irregular, sinusoidal spaces. All three patients had local recurrence, but all were tumor-free at latest follow-up. PMID- 2982255 TI - Cytologic criteria for the diagnosis of pancreatic carcinoma. AB - Fine-needle aspiration and endoscopic aspiration of pancreatic cells permit the diagnosis of pancreatic carcinoma and avoid the complications and morbidity of pancreatic biopsy. In this study, the accuracy of fine-needle and endoscopic aspiration were compared, and cytologic criteria for pancreatic carcinoma were sought. Pancreatic cytologic preparations from 79 patients, including 39 fine needle aspirates and 48 endoscopic aspirates, were retrospective reviewed. When compared with definitive tissue diagnosis or clinical course, fine-needle aspiration had a sensitivity for pancreatic carcinoma of 79%. Endoscopic aspiration of pancreatic secretions had a sensitivity of only 33%. There was a single falsely suspicious fine-needle aspirate, but there were no false positive diagnoses when using either collection technic. Seventeen cytologic features were examined to determine cytologic criteria of malignancy. The presence of disoriented or crowded cells in three-dimensional groups, and extreme nuclear enlargement combined with nuclear contour irregularity were the best criteria for pancreatic malignancy. PMID- 2982256 TI - Persistent fetal circulation. Neurodevelopmental outcome. AB - Forty children who had persistent fetal circulation (PFC) were followed up for one to four years. At the most recent examination, 16 (40%) were normal, eight (20%) had neurosensory hearing loss, three (7.5%) were profoundly impaired, six (15.0%) had suspect or abnormal results of neurologic examinations exclusive of hearing loss, three (7.5%) had speech impairment and normal hearing, and one had a delay in motor development. The remaining three had clinically suspect speech. Neurosensory hearing loss correlated only with having a mother with insulin dependent diabetes, hand-to-mouth facility on the Brazelton scale, and eight month neurologic status. Earlier diagnosis of hearing loss may be facilitated by testing of neonatal auditory behavioral responses, neurologic status at eight months, and by attention to parental concern about abnormal speech development. Neonatal and infancy auditory screening are recommended in children who have PFC in the neonatal period. PMID- 2982257 TI - Administrating incompatible drugs by a retrograde intravenous infusion system. AB - Use of a retrograde infusion system for concurrent intravenous administration of two incompatible drugs separated by a barrier fluid was studied. Four different barrier fluids (5% dextrose injection, 10% dextrose injection, 0.9% sodium chloride injection, and sterile water for injection) were used to separate sodium bicarbonate and calcium chloride. The primary infusion was 5% dextrose injection. The delivery system was visually inspected for formation of precipitate (calcium carbonate) upon addition of the two drug solutions and 2 ml of the barrier fluid. If precipitation occurred, the procedure was repeated, increasing the volume of barrier fluid incrementally until no precipitate could be seen. If no precipitate was seen with the initial 2 ml of barrier fluid, the volume of barrier fluid was decreased until precipitation occurred. These procedures were repeated for each barrier fluid at flow rates of 5-20 ml/hr for 90 minutes. The minimum volume of barrier fluid needed to prevent precipitate formation was determined in triplicate at each flow rate. The minimum volume of barrier fluid that prevented precipitate formation was approximately 2.0 ml. This volume did not differ significantly by barrier solution type or by flow rate. Sodium bicarbonate and calcium chloride can be administered concurrently by the retrograde intravenous method without visual incompatibility when separated by greater than or equal to 2 ml of barrier fluid, and this method can probably be used for administration of other potentially incompatible drugs. PMID- 2982259 TI - Early gastric cancer. Clinicopathologic study. AB - The incidence and clinicopathologic features of early gastric cancer encountered among surgical specimens from gastric resections for carcinoma in a recent three year period, 1977 to 1979, at the Mallory Institute of Pathology were studied and compared with those of a pre-endoscopic period 10 years earlier, 1967 to 1969. It was found that early gastric cancer now comprises a greatly increased proportion of lesions leading to gastric resection, mainly as a result of endoscopy and biopsy of gastric ulcers of benign appearance. In the recent period, there were six early gastric cancers in a total of 22 gastric resection specimens compared with one in 27 gastric resections performed for carcinoma in the pre-endoscopy period. Five of the six patients in the recent period are alive without evidence of disease four to five years following surgical resection. The single patient in the earlier period died postoperatively. Applying the classification of the Japanese Endoscopic Society, there were three depressed or ulcerated lesions (type IIc or III), three elevated or polypoid lesions (type I or IIa), and a single flat lesion (type IIb). All three ulcerated lesions were interpreted as benign peptic ulcers on conventional upper gastrointestinal studies. Findings on endoscopic biopsy were positive in all cases (six of six). Although not encountered frequently in the United States, early gastric cancer, nonetheless, appears to be indistinguishable from the disease as it is described in Japan in terms of its pathologic morphology, growth patterns, coexistent or related lesions of the stomach, and curability by surgical resection. If early gastric cancer is to be recognized more frequently, knowledge of the disease and a high index of suspicion on the part of physicians are essential. PMID- 2982258 TI - Immune system in rheumatoid arthritis: possible implications in neoplasms. AB - Although the immune system in rheumatoid arthritis is generally in a state of hyperactivity, deficient function can be demonstrated in certain specific parts of the system. This has been most clearly demonstrated in studies of the response of blood lymphocytes from demonstrated in studies of the response of blood lymphocytes from patients with rheumatoid arthritis to in vitro infection by the Epstein-Barr virus. The virus normally infects B cells promoting proliferation. This results in the establishment of permanent lines of infected B cells more commonly than normal in rheumatoid arthritis. This condition is associated with decreased production of lymphokines, interleukin-2, and gamma interferon. Similar in vitro problems in immune responses to other antigens have also been described. Although natural killer cells are scarce among synovial cells, they are present in normal numbers in the blood. Whether these select immunodeficiencies make patients with rheumatoid arthritis more prone to neoplasms, particularly lymphomas, is a question that has not been investigated until now. Nor has it been determined whether the immune abnormalities in rheumatoid arthritis make patients more prone to neoplasms from the therapeutic agents used in their treatment. PMID- 2982260 TI - Pancreatic islet cell carcinoma with hypercalcemia. Primary hyperparathyroidism or humoral hypercalcemia of malignancy. AB - A 60-year-old woman presented with hypercalcemia and was found to have metastatic pancreatic islet cell carcinoma. Although clinical features were very suggestive of hyperparathyroidism, her parathyroid hormone levels were not elevated and no abnormal parathyroid tissue was detected by thallium-technetium or computed tomographic scanning techniques. Her hypercalcemia appeared to be due to a humoral factor--distinct from parathyroid hormone--that mimics the action of parathyroid hormone almost exactly. The various tools that may be used to differentiate primary hyperparathyroidism from the humoral hypercalcemia of malignancy are reviewed. PMID- 2982261 TI - Diabetes insipidus from metastatic oat cell carcinoma: recovery after brain irradiation. AB - A hypothalamic metastasis was demonstrated by computed tomography in a 71-year old patient, with previously unknown small cell lung cancer, who presented with diabetes insipidus and biological signs of hypothyroidism and hypogonadism. Brain irradiation resulted in resolution of polyuria, elevation of thyroid hormones, improvement of pituitary responsiveness to hypothalamic releasing hormones, and complete disappearance of contrast-enhanced suprasellar metastasis. PMID- 2982262 TI - Odontoonychodysplasia with alopecia: a new pure ectodermal dysplasia with probable autosomal recessive inheritance. AB - We report on two Brazilian sisters who have a probably autosomal recessive ectodermal dysplasia of trichodysplasia, dental anomalies, onychodystrophy, skin alterations, and other findings. This appears to be the first full report of this condition for which we propose the name odontoonychodysplasia with alopecia. PMID- 2982263 TI - Syndrome of polydactyly, conical teeth and nail dysplasia. PMID- 2982264 TI - The association of sexually transmitted diseases with cervical intraepithelial neoplasia: a case-control study. AB - Thirty-three women with histologically confirmed cervical intraepithelial neoplasia (grades I to III, with one case of microinvasive carcinoma) and 54 women without evidence of the disease were prospectively studied to determine the relationship of genital infection to cervical neoplasia. Demographic and sexual data for patients and control subjects were collected, with standardized clinical and colposcopic evaluation by means of predefined diagnostic categories. Cultures from the cervix were examined for herpes simplex virus, cytomegalovirus, Chlamydia trachomatis, and Neisseria gonorrhoeae. Human papilloma virus infection was identified by characteristic changes of koilocytosis in cytologic or histopathologic specimen. Cultures from the vagina were evaluated for Gardnerella vaginalis, Trichomonas vaginalis, Ureaplasma urealyticum, Mycoplasma hominis, Candida albicans, and other yeasts. Separate Gram strains were prepared from endocervical secretions and from vaginal secretions. More lifetime sexual partners, larger area of transformation zone, evidence of human papilloma virus infection, and altered vaginal flora were observed in women with cervical intraepithelial neoplasia. The association of human papilloma virus infection and altered vaginal flora with cervical intraepithelial neoplasia was independent of sexual experience. PMID- 2982265 TI - Functional zonation of the midgestation human fetal adrenal cortex: fetal versus definitive zone use of progesterone for cortisol synthesis. AB - Studies of human fetal adrenal function and its control have revolved mainly around the remarkable capacity of the unique fetal zone of this gland to elaborate dehydroepiandrosterone sulfate. Another important function of the fetal adrenal, however, is its production of cortisol. Because the human fetal adrenal is deficient in 3 beta-hydroxysteroid dehydrogenase activity, cortisol has been thought to be formed from circulating progesterone. To further investigate this hypothesis, cortisol production by separated fetal and definitive zones of the midgestation human fetal adrenal in organ culture has been examined in the absence and presence of varying concentrations of progesterone and adrenocorticotropic hormone. Cortisol was measured by radioimmunoassay. In the absence of progesterone, cortisol production by both zones increased gradually over time in culture in response to adrenocorticotropic hormone. In the presence of progesterone, cortisol production by the definitive zone was unchanged. In contrast, the response of the fetal zone to progesterone was immediate: cortisol production increased significantly and remained high throughout the culture period. These results suggest a greater capacity of the fetal zone to utilize progesterone for cortisol production and are consistent with morphologic evidence that the active zone of the midgestation human fetal adrenal is the fetal zone, possessing not only the enzyme activity necessary for dehydroepiandrosterone sulfate production but, except for 3 beta-hydroxysteroid dehydrogenase, that for cortisol production as well. PMID- 2982266 TI - Effect of naloxone on menopausal flushes, skin temperature, and luteinizing hormone secretion. AB - The effect of naloxone (1.4 mg/hr for 3 hours) on subjectively experienced menopausal flushes, skin temperature, and luteinizing hormone secretion was investigated in seven women in a double-blind, saline-controlled, crossover study. Naloxone had no effect on the number of subjective flushes, episodic skin temperature elevation, luteinizing hormone pulses, variability of luteinizing hormone secretion, or total luteinizing hormone secretion. This study suggests that a naloxone-sensitive opioid mechanism is not active in modulating luteinizing hormone secretion in the postmenopausal woman and that opioid receptor blockade is not effective in altering the frequency of menopausal flushes. PMID- 2982267 TI - Controversies in Rh prophylaxis. Who needs Rh immune globulin and when should it be given? AB - A summary of the current recommendations for administration of Rh immune globulin after several reproductive events is presented. The risks of failure to administer Rh immune globulin after delivery or abortion, amniocentesis, and massive transplacental hemorrhage are described. PMID- 2982268 TI - Collagen sponge as vaginal contraceptive barrier: critical summary of seven years of research. AB - Extensive testing of collagen sponge as a vaginal contraceptive (mechanical and chemical) showed that the original expectations regarding the safety, convenience, and efficacy were not met. The collagen sponge was tested both as a cylinder and as a diaphragm and used as such or impregnated with spermicidal detergent or with zinc salt. The collagen sponge must be larger than 6 cm in diameter in order to serve as a mechanical barrier that will not be dislodged during physical activity. This creates problems with the ease of insertion and with the partners' awareness of the barrier. When the collagen sponge containing ejaculate is left in the vagina greater than 48 hours, it develops an offensive odor. The original acidity of the collagen sponge (pH 3.5, 0.1 mol/L) is soon neutralized by the large volume of alkaline vaginal secretions. In vitro studies showed that up to 10 mg of nonoxynol 9 per milliliter of growth medium did not inhibit the growth of Staphylococcus aureus. These effects, as well as the large surface area of the resilient sponge, present a potential risk for growing staphylococci within the collagen sponge. The capacity of the collagen sponge to absorb a large volume of cervical and vaginal fluid produced two symptoms that were annoying to the volunteers: an awareness of either vaginal dryness during intercourse or, conversely, saturation of the sponge from the vagina. Postcoital studies showed viable spermatozoa in the cervical mucus in 25% of the tests with the nonmedicated cylindrical sponge but in only 6% of tests with the sponge containing nonoxynol 9. The results of clinical trials conducted at four centers support the view that collagen sponge as a vaginal contraceptive barrier method is inconvenient to both partners, not effective enough to compete with present methods of vaginal contraception, and possibly might be unsafe because of the capacity to grow bacteria. Despite the negative end result of this goal-oriented research, we believe that our studies have contributed to a better understanding of vaginal physiologic features, the safety and effectiveness of spermicidal detergents, and the mechanisms of vaginal malodor. Although the acceptability study showed some advantages of the collagen sponge over the rubber diaphragm, the overall acceptability of the collagen sponge diaphragm was no better than that of the rubber diaphragm. For all these reasons, including the possible risk of an increased incidence of toxic shock syndrome, we have discontinued further testing of either type of collagen sponge as a vaginal barrier method. PMID- 2982269 TI - Beta-adrenergic receptors in human myometrium during pregnancy: changes in the number of receptors after beta-mimetic treatment. AB - The concentration of available beta-adrenoceptors has been studied in the myometrium of women treated with terbutaline for premature uterine contractions and in an untreated control group. Myometrial strips were taken at cesarean section from the lower uterine segment and the uterine fundus. The concentration of beta-adrenoceptors was determined with a radioligand binding assay. In untreated women we found no difference in the concentration of beta-adrenoceptors in the uterine fundus compared to that in the lower uterine segment. The cyclic adenosine monophosphate production after beta-adrenoceptor agonist stimulation in vitro was equal in both locations. In the terbutaline-treated women, the binding site concentrations in both the fundus and lower uterine segment were lower compared to those in the control group. The decrease was most pronounced in the fundus where receptor concentration was only half that found in the control group. The results suggest that treatment with beta-mimetics causes a desensitization in the beta-adrenoceptor system of human myometrium during pregnancy. This desensitization may partly explain the limited duration of the relaxant effect of beta-mimetics often seen in the treatment of preterm labor. PMID- 2982270 TI - Characterization of two types of crystalloids in pleomorphic adenomas of minor salivary glands. A light-microscopic, electron-microscopic, and histochemical study. AB - Crystalloids have been previously described in salivary gland tumors. In order to ascertain the incidence of these structures, the authors reviewed a series of 294 minor salivary gland tumors. One hundred thirty pleomorphic adenomas were identified, and 6 of these contained crystalloids. No crystalloids were found in other benign or malignant salivary gland tumors. These six file cases and a recent seventh case containing crystalloids were studied by light and electron microscopy and with histochemistry. Two types of crystalloids were found. One case contained previously described tyrosine-rich crystalloids, and the other six contained crystalloids composed of radially arranged collagen fibers. Both types of crystalloids are further characterized and discussed. PMID- 2982272 TI - X-linked hypophosphatemia (familial or sex-linked vitamin-D-resistant rickets). X linked hypophosphatemic (Hyp) mice. PMID- 2982271 TI - Histologic, immunofluorescence, and ultrastructural study of malignant islet-cell tumors of the pancreas induced in hamsters by BK human papovavirus. AB - Histologic, immunofluorescence and ultrastructural studies were performed in 17 cases of pancreatic carcinomas induced by the BK virus in Syrian hamsters, a unique model of experimentally induced malignant islet cell tumors. The tumors were composed of small, poorly differentiated cells mostly arranged in a trabecular structure. By immunofluorescence all four islet cell types were found in the tumors, though with different frequency. Insulin cells were present in 16 cases, glucagon cells in 11, somatostatin cells in 7, PP cells in 6. Thirteen tumors contained more than one cell type. Insulin cells were the most frequent cell type in 13 cases, and glucagon cells predominated in 1 case. Insulin containing cells usually occupied a central position within tumor-cell aggregates, while the other cell types were mostly located in a peripheral position, a distribution reminiscent of that seen in normal islets. Gastrin and calcitonin immunoreactivities were not observed. Immunoreactive cells were more abundant in tumors with trabecular structure. Argyrophil cells revealed by the Grimelius method often exceeded the cumulative number of immunoreactive cells in the same tumor, which suggests that there were additional cell types. Multiple cell types were also found in liver metastases. Ultrastructurally most neoplastic cells were poorly granulated. The occurrence of many damaged cells suggests hormone leakage, which may account, at least in part, for the deregulated hormone release from the tumors. PMID- 2982273 TI - Parathyroid hormone contributes to volume expansion-induced inhibition of proximal reabsorption. AB - Volume expansion inhibits the proximal reabsorption of water, bicarbonate, and chloride. The present work tested a hypothetical role of parathyroid hormone (PTH) in the expansion effect. We studied 19 Sprague-Dawley rats during a plasma replete euvolemic state and following 10% body wt colloid-free expansion. In group I, six intact rats, volume expansion decreased plasma ionized calcium concentration ([Ca2+]P) from 2.28 +/- 0.06 to 2.11 +/- 0.04 meq/liter and increased nephrogenous cAMP (NcAMP) from 29 +/- 5 to 66 +/- 10 pmol X min-1 X g kidney wt-1. In group II, six acutely thyroparathyroidectomized (TPTX) rats, [Ca2+]P also fell from 2.18 +/- 0.08 to 1.80 +/- 0.08 meq/liter but NcAMP did not rise significantly (9 +/- 3 vs. 17 +/- 5 pmol X min-1 X g kidney wt-1). These data strongly suggest that stimulation of PTH activity occurred during expansion in intact animals. In group III, seven TPTX rats, volume expansion inhibited proximal reabsorption of total CO2 by 11%, of chloride by 24%, and of water by 19%. Volume expansion-induced reduction in bicarbonate, chloride, and water reabsorption was smaller in TPTX than in intact rats previously studied. We conclude that volume expansion inhibits proximal reabsorption in part by decreasing the active transcellular NaHCO3 and NaCl transport secondary to stimulation of PTH activity. PMID- 2982274 TI - Renal alpha-adrenoceptors and sodium excretion in the dog. AB - The purpose of this study was to characterize the age-dependent role of alpha adrenergic activity on renal vascular resistance (RVR) and renal blood flow (RBF) and the role of alpha 2-receptors in sodium transport. Intrarenal infusion of phentolamine in hydropenic anesthetized pups and adult dogs increased RBF and glomerular filtration rate (GFR) only in pups. The effect was most marked in the youngest pups (15.67 +/- 0.67 days). At 1.0 micrograms X kg-1 X min-1, phentolamine increased RBF from 1.38 +/- 0.04 to 1.64 +/- 0.04 ml X min-1 X g kidney wt-1 and GFR from 0.19 +/- 0.01 to 0.23 +/- 0.01 ml X min-1 X g kidney wt 1. Absolute (UNaV) and fractional (FENa) sodium excretions increased in all animals, but mean percent increases were greatest in adult dogs. Intrarenal yohimbine infusion in adult dogs (10-100 micrograms X kg-1 X min-1) produced a dose-related increase in UNaV and FENa without affecting RBF and GFR. UNaV increased from 0.22 +/- 0.05 to 0.54 +/- 0.12 mueq X min-1 X g kidney wt-1 and FENa increased from 0.32 +/- 0.05 to 0.63 +/- 0.06% at the dose of 100 micrograms X kg-1 X min-1. These studies confirm a modest role for alpha-adrenoreceptors in the high RVR characteristic of newborn pups and provide evidence for a role of alpha 2-adrenoceptors in the renal transport of sodium; the extent of the contribution of renal alpha-adrenergic system could not be tested in this experiment. PMID- 2982275 TI - ATP and cAMP system in the in vitro response of microdissected cortical tubules to PTH. AB - Responsiveness of proximal convoluted tubule (PCT) and distal convoluted tubule (DCT) microdissected from mouse kidney to PTH, in terms of cAMP accumulation and stimulation of adenylate cyclase, was examined. In both PCT and DCT, the cell free adenylate cyclase was stimulated at least 10-fold by the same dose (10 U/ml) of PTH, and activity of cAMP phosphodiesterase was about 80% higher in DCT than in PCT. In intact tubules, while the incubation with PTH increased cAMP content in DCT more than 10-fold, it failed to increase the cAMP levels in PCT. To explain discrepancies between cell-free and intact cell incubations, ATP content in microdissected tubules was determined with use of a microbioluminescence assay. ATP content in PCT (4.0 +/- 1.3 fmol/mm, n = 30) was dramatically lower than ATP content of DCT (376.8 +/- 54.3 fmol/mm, n = 25). Incubation with 1 microM rotenone reduced markedly (delta -98%) the ATP content in DCT. In DCT, with ATP depleted by 1 microM rotenone, PTH failed to increase the cAMP, although 1 microM rotenone did not inhibit the adenylate cyclase activity. When 0.1 mM of 1-methyl-3-isobutylxanthine (MIX) was added to the incubation medium, PTH caused a marked elevation in tubular cAMP in PCT and to even a greater degree in DCT. Present results show that various segments of microdissected tubules differ greatly in their ability to maintain adequate ATP levels for cAMP generation in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982276 TI - Effect of aluminum on the renal handling of phosphate in the rat. AB - The effects of aluminum (Al) on the renal handling of phosphate (Pi) were studied by clearance techniques in the presence and absence of endogenous parathyroid hormone (PTH) and after Pi infusion. In all groups, a 1-h control period was followed by three 1-h experimental periods in which controls continued to receive saline and experimental animals received Al. Glomerular filtration rate, urine flow rate, and plasma sodium (Na) were not significantly changed between periods in any group. In the presence of endogenous PTH, the fractional excretion of phosphate (FEPi) increased significantly after 3 h of Al infusion (6.1 +/- 0.9 to 15.0 +/- 1.8%, P less than 0.05, n = 8), but not in controls (n = 7), while plasma calcium and Pi decreased, suggesting a PTH effect. However, in acutely thyroparathyroidectomized (TPTX) rats the FEPi also increased significantly after 3 h of Al infusion (4.3 +/- 3.0 to 10.6 +/- 4.2%, P less than 0.05, n = 7), but not in controls (n = 6). In TPTX rats infused with Pi where plasma Pi was increased to 10 mg/dl, FEPi increased significantly after 2 and 3 h of Al (7.0 +/ 1.4 to 15.5 +/- 2.1 to 16.9 +/- 2.2%, P less than 0.01, n = 15), but not in controls (n = 8). In this group, changes in FEPi were accompanied by a small but significant increase in FENa but not urinary cAMP. Blood pH was not significantly different between saline and Al-infused rats. These studies indicate that Al infusion inhibits renal Pi reabsorption by a mechanism independent of PTH, blood pH, or cAMP. PMID- 2982277 TI - Proton/hydroxyl permeability of proximal tubule brush border vesicles. AB - The net H+/OH- permeability of rabbit renal proximal tubule brush border membrane vesicles was determined by measuring the rate of collapse of preformed pH gradients using acridine orange. The membranes were voltage clamped using valinomycin and [K+]in = [K+]out. Internal buffer capacity was determined by titration of lysed vesicles and by titration of measured Na+/H+ exchange rates with exogenously added buffers. Both methods revealed an intravesicular buffer capacity of 125-135 mM/pH unit at pH 6.0 and 20 degrees C. Using this buffer capacity, the net H+/OH- permeability was found to be 5 X 10(-3) cm/s in brush border vesicles prepared by Mg2+ aggregation. The rate of collapse of pH gradients in brush border vesicles prepared by sucrose density gradient centrifugation was virtually identical to the rate in vesicles prepared with Mg2+, indicating that the high H+/OH- permeability was not an artifact of Mg2+ preparation. Activation energy of the H+/OH- permeability pathway was 4.9 kcal/mol, whereas activation energy of the Na+/H+ antiporter was 11.4 kcal/mol. Since the rate of H+/OH- diffusion was not affected by amiloride, it is concluded that H+/OH- permeate through brush border membranes by a pathway separate from the Na+/H+ antiporter. This pathway is not inhibited by dicyclohexylcarbodiimide at concentrations up to 2 mM but is inhibited by 0.2-5 mM p chloromercuribenzenesulfonate, suggesting the presence of a sulfhydryl group in the pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982278 TI - Inhibition of sarcolemmal Na+-K+-ATPase by palmitoyl carnitine: potentiation by propranolol. AB - Native sarcolemma (SL) from adult canine cardiac myocytes (Na+-K+-ATPase activity 74.2 +/- 3.0 mumol X mg-1 X h-1) was preincubated (10 min, 37 degrees C, pH 7.2) with 1) 20-600 microM palmitoyl carnitine, 2) 250 nM-2.5 mM propranolol, or 3) 20 600 microM palmitoyl carnitine plus propranolol at various concentrations (0.0, 0.025, 0.25, 0.5, 1.0, and 2.5 mM); after preincubation, Na+-K+-ATPase activity was assayed. Palmitoyl carnitine alone (series 1) had no effect on ATPase activity over the range of 20-400 microM but was inhibitory (30%) at 600 microM. Propranolol alone (series 2) did not alter ATPase activity at any concentration. When SL membranes were exposed to both palmitoyl carnitine and propranolol (series 3), a dose-dependent inhibition of ATPase activity was observed. The inhibitory effect was not reversed by 3.0% bovine serum albumin. Propranolol concentrations greater than 0.025 mM significantly inhibited the activity of SL exposed to palmitoyl carnitine (above 150 microM). Palmitoyl carnitine and propranolol do not have to be added simultaneously to produce combined inhibition. Activity was inhibited 50% when SL were pretreated with 100 microM palmitoyl carnitine followed by addition of 2.5 mM propranolol no inhibition occurred if preincubation conditions were reversed. Thus exposure of SL to propranolol and reported physiological levels of palmitoyl carnitine leads to irreversible inhibition of the Na+-K+-ATPase, which may be due to the combined membrane-perturbant actions of these amphipathic agents. PMID- 2982279 TI - Properties of renobulbar afferent fibers in rats. AB - Myelinated renobulbar sensory fibers project directly from the kidney to the dorsomedial medulla. They can be activated by punctate stimulation of the kidney. In these experiments we studied the physiological modality and the intrarenal locations of the receptors for renobulbar fibers. Renobulbar fibers were identified in recordings from the renal nerves of anesthetized rats. Although nonmyelinated renal afferent fibers were activated by increased intrarenal pressure and by chemical stimuli, no renobulbar fiber activated by punctate mechanical stimulation was activated by these other stimuli. Occasionally single fibers could be activated by punctate stimulation of two or more sites on the surface of the kidney, and responses from each site could be selectively blocked with a local anesthetic. Renobulbar fibers could be activated by electrical stimulation of discrete regions deep within the renal parenchyma. Increased intrarenal pressure increased the sensitivity of receptors to punctate stimulation but not to electrical stimulation of the kidney. We conclude that myelinated renal afferent fibers in the rat respond to external mechanical stimuli, lie deep within the renal parenchyma, and occasionally have more than one receptor. PMID- 2982280 TI - Attenuation of postexercise ketosis in fasted endurance-trained rats. AB - Endurance-trained animals and human subjects have been reported to exhibit a lesser degree of postexercise ketosis than nontrained controls. We have studied the mechanism of this adaptation. Trained (2 h/day, 6 wk) and nontrained rats were fasted overnight and then run at 16 m/min up a 15% grade for 90 min. Trained rats had lower blood 3-hydroxybutyrate during exercise and during a 90-min postexercise period than nontrained rats. Liver malonyl coenzyme A (CoA), carnitine, and glycogen were not significantly different in the two groups at any time during and after exercise. Therefore these factors cannot be responsible for the difference in ketonemia. Plasma free-fatty acids and hepatic adenosine 3',5' cyclic monophosphate were elevated in nontrained rats with respect to trained rats. These two differences could conceivably be responsible for a different ketogenic rate. In addition, 3-ketoacid CoA transferase activity of gastrocnemius muscle was increased by training. The increase in ketone oxidizing enzymes of muscle may also be partially responsible for the training-induced attenuation of postexercise ketonemia in these fasted rats. PMID- 2982281 TI - Cultured Sertoli cell-mediated FSH stimulatory effect on Leydig cell steroidogenesis. AB - To determine the precise role of Sertoli cells in the stimulating effects of follicle stimulating hormone (FSH) on Leydig cell activity, porcine purified Leydig and Sertoli cells were cultured separately or together in a chemically defined medium in the absence or presence of porcine, FSH 50 ng/ml. Leydig cell activity was evaluated using two parameters: human chorionic gonadotropin (hCG) binding sites; and hCG-stimulated cAMP production and testosterone secretion. First, it was found that FSH increases Leydig cell activity in crude Leydig cell preparations (40-60% of Leydig cells), whereas it exerts no effect on purified Leydig cells (greater than 90% of Leydig cells). Second, FSH stimulates the activity of Leydig cells cocultured with Sertoli cells, whereas it remains without effect on purified Leydig cells cultured alone. This stimulating effect of FSH on Leydig cell activity is dependent on the Sertoli cell number in the coculture. These data 1) show that the stimulating effect of FSH on Leydig cell function is mediated by Sertoli cells and 2) support the concept of local control of Leydig cell function originating from Sertoli cells. PMID- 2982282 TI - Reduced responsiveness of glomerulosa cells after prolonged stimulation with angiotensin II. AB - The purpose of this study was to examine whether sustained exposition to angiotensin modifies the responsiveness of adrenal glomerulosa cells when extra adrenal factors are eliminated. Isolated rat glomerulosa cells were stimulated for 6 h in a superfusion system with angiotensin II or potassium. Their responsiveness to angiotensin II, potassium, and corticotropin (ACTH) was examined before and after the superfusion. Stimulation of the cells during the superfusion with angiotensin II or with potassium reduced their responsiveness to all three stimuli. The steroid synthesis inhibitor aminoglutethimide, applied during the superfusion, overcame the effect of potassium but failed to influence that of angiotensin. This suggests that the reduced responsiveness after stimulation with potassium is related to the increased steroid production whereas the action of angiotensin is independent of that. The results establish the existence of desensitization to angiotensin in the absence of modifying extra adrenal factors. PMID- 2982283 TI - cAMP metabolism and lipolysis in brown adipocytes of hamsters consuming a cafeteria diet. AB - Feeding animals cafeteria diets causes increased sympathetic activity to brown adipose tissue and this is believed to be responsible for the concomitant activation of thermogenesis. Because chronic catecholamine stimulation in other systems leads to a desensitization of beta-adrenergic receptors, we examined lipolysis and cAMP production in brown adipocytes of hamsters eating cafeteria diets for evidence of diminished beta-adrenergic responses. Basal cAMP levels were similar in chow- and cafeteria-fed hamsters. However, adipocytes from overfed animals formed less cAMP in response to isoproterenol than those of control animals. Isoproterenol-stimulated adenylate cyclase activity was similarly decreased in membrane preparations from cafeteria-fed hamsters. However, when the diterpene forskolin was used, equal amounts of cAMP were formed in cells and membrane preparations from control and overfed animals. In contrast to the reduced responses of the cAMP system to isoproterenol stimulation observed in overfed hamsters, isoproterenol-stimulated lipolysis was greater in cells from overfed animals than in cells from control animals. These results are consistent with a desensitization of the adenylate cyclase system in brown adipocytes occurring during chronic hyperphagia. Because eating cafeteria diets has been reported to increase sympathetic activity to brown fat depots, the apparent desensitization of brown adipocytes observed in this study may result from a persistent stimulation of the brown fat with norepinephrine in vivo. Our data also suggest the existence of mechanisms that preserve lipolysis in the face of low cAMP levels. PMID- 2982284 TI - Water deprivation upregulates angiotensin II receptors in rat anterior pituitary. AB - Angiotensin II (ANG II) receptors were quantitated in pituitary glands of individual male Long Evans rats by autoradiography after incubation of 8-microns thick pituitary sections with 125I-[Sar1]ANG II. Rat anterior pituitary had a single class of high-affinity saturable ANG II receptors with a Bmax of 1,360 +/- 109 fmol/mg protein and a Ka of 0.510 +/- 0.03 X 10(9) M-1. Five days of water deprivation produced a marked increase in the number of anterior pituitary ANG II receptors (Bmax: 2,428 +/- 233 fmol/mg protein, a 79% increase, P less than 0.001) and a decrease in affinity for the ligand (Ka: 0.337 +/- 0.01 10(9) M-1, a 34% decrease, P less than 0.05). Our results suggest a role for anterior pituitary ANG II receptors in the regulation of fluid and electrolyte metabolism in the rat. PMID- 2982285 TI - Parathyroid hormone and dibutyryl cAMP inhibit Na+/H+ exchange in renal brush border vesicles. AB - Parathyroid hormone (PTH) and cAMP inhibit sodium, water, and bicarbonate reabsorption in the proximal tubule. We wished to determine whether these agents directly inhibit proximal tubular Na+/H+ exchange. A suspension of rabbit proximal tubules was prepared by enzymatic digestion and Ficoll gradient centrifugation. Oxygen consumption at 37 degrees C was stable over 60 min, averaged 20 nmol X mg protein-1 X min-1, and was inhibited 60% by ouabain. Over 96% of cells excluded trypan blue. From this suspension, brush border membrane vesicles were isolated. The vesicles were enriched 12.7 times in alkaline phosphatase relative to a cortical homogenate and demonstrated pH gradient stimulated, amiloride-sensitive Na+/H+ countertransport and sodium-phosphate and sodium-D-glucose cotransport. When the tubule suspension was exposed to PTH or dibutyryl cAMP, the activity of Na+/H+ countertransport in the resultant brush border vesicles was inhibited. Neither PTH nor dibutyryl cAMP affected the amiloride-insensitive component of sodium transport or sodium-phosphate or sodium D-glucose cotransport. The effect of PTH on Na+/H+ counter-transport could not be explained by an alteration in fluidity of the brush border membrane. These experiments demonstrate that PTH and dibutyryl cAMP directly inhibit Na+/H+ countertransport in the brush border membrane of the rabbit proximal tubule. PMID- 2982286 TI - Activation of hypothalamo-hypophysial-interrenal system by urophysectomy in goldfish. AB - Plasma cortisol, pituitary adrenocorticotropin and arginine vasotocin (AVT) contents, and hypothalamic urotensin I (UI) and AVT contents were determined in urophysectomized goldfish (1 wk) with and without dexamethasone injections. Urophysectomy produced marked increases in plasma cortisol. These marked increases could not be observed in urophysectomized fish that received dexamethasone injections. These findings suggest that the hypothalamo-pituitary interrenal axis is activated after urophysectomy. This activation may be due to the enhanced secretion of U I from the goldfish hypothalamus, suggesting that U I may be a fish corticotropin-releasing factor. PMID- 2982288 TI - ACTH and cortisol levels in healthy probands and psychiatric patients following the dexamethasone suppression test. PMID- 2982287 TI - Cl- -HCO3- -stimulated ATPase in intestinal mucosa of Aplysia. AB - The serosa negative transepithelial potential difference across Aplysia intestine is generated by a Na+-independent, active electrogenic Cl- absorptive mechanism. In an attempt to clarify the Cl- absorptive mechanism an anion-stimulated ATPase was prepared from plasma membranes from Aplysia enterocytes utilizing differential centrifugation and sucrose density gradient techniques. ATPase activity, which could be activated by either Cl- or HCO3-, was found in the plasma membrane fraction. Maximal anion-ATPase activity was achieved with either 25 mM Cl- or 25 mM HCO3-. The apparent Km for Cl- activation of the ATPase was 10.3 mM, whereas apparent Km for HCO3- was 9.7 mM. ATP was the most effective nucleotide substrate for both HCO3- and Cl- -ATPase activities, whereas optimum pH for both activities was 7.8. These enzyme activities were inhibited more than 30% by thiocyanate (10 mM). Acetazolamide and vanadate were also found to strongly inhibit both Cl- and HCO3- -ATPase activities, whereas 10 microM 4 acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, 1 mM furosemide, or 1 mM ouabain had little or no effect. These results are consistent with the hypothesis that the active Cl- transport mechanism in Aplysia intestine could be a Cl- -HCO3 -stimulated ATPase found in the enterocyte plasma membrane. PMID- 2982289 TI - In vitro cultivation of the exoerythrocytic stage of Plasmodium berghei in irradiated hepatoma cells. AB - Growth of cultures of human hepatoma cells was inhibited by exposure to doses of gamma irradiation as low as 1,000 rad., and the monolayers remained viable for up to 35 days. Irradiated cells were at least as susceptible to Plasmodium berghei sporozoite invasion as non-irradiated cells, and supported the entire exoerythrocytic cycle producing more infectious merozoites. Irradiated cultures may have use for culture of human malarias, and drug studies requiring synchronous cultures. PMID- 2982290 TI - Ectopic ACTH syndrome. Diagnostic and therapeutic aspects. AB - Twenty-five patients were identified with non-pituitary, nonadrenal ACTH secreting tumors (bronchial carcinoid, bronchial small cell carcinoma, pancreatic islet cell carcinoma, medullary thyroid carcinoma, thymic carcinoids, metastatic adenocarcinoma, and pancreatic cystadenoma). Clinical features were weakness, hypertension, cushingoid appearance, peripheral edema, personality disorders, and hyperpigmentation. Biochemical features were a markedly increased urinary free cortisol level (all patients), hypokalemia (71 percent of patients), and an elevated ACTH level (72 percent of patients). Surgical therapy consisted of bilateral total adrenalectomy (56 percent of patients). Twelve percent underwent transsphenoidal hypophysectomy and 36 percent had excision of their tumor. No surgical therapy was undertaken in 28 percent. Bilateral total adrenalectomy in patients with a slow-growing malignancy or an unknown tumor secreting ACTH is beneficial in relieving symptoms and prolonging life. Excision of nonmalignant ACTH-producing tumors yields an excellent long-term prognosis. PMID- 2982291 TI - Simulated metastatic carcinoma after breast reconstruction. AB - In two patients who had undergone mastectomies and reconstruction for breast carcinoma, suspicious lesions were noted on chest roentgenograms. Subsequent studies revealed the lesions to be shadows from the reconstructed nipples. The most frequently employed techniques for nipple reconstruction involve the grouping and overlapping of multiple local flaps which results in a round radiodensity on a roentgenogram. Patients with suspicious roentgenographic lesions after breast reconstruction should have oblique films taken to localize the lesion. PMID- 2982292 TI - Case study: a child with chronic pulmonary infiltrates. Part one. PMID- 2982293 TI - Role of insects in the transmission of bovine leukosis virus: potential for transmission by stable flies, horn flies, and tabanids. AB - The ability of stable flies (Stomoxys calcitrans), horn flies (Haematobia irritans), and tabanids (Diptera: Tabanidae) to transmit bovine leukosis virus (BLV) was investigated. Stable flies and horn flies were fed on blood collected from an infected cow, and the flies' mouthparts were immediately removed, placed in RPMI-1640 medium, ground, and inoculated into sheep and calves. Infection of sheep occurred with mouthparts from as few as 25 stable flies or 25 horn flies. However, sheep were not infected when removal of stable fly mouthparts was delayed greater than or equal to 1 hour after blood feeding. Infection of calves occurred after inoculation of mouthparts removed immediately after feeding from as few as 50 stable flies or 100 horn flies. Infected blood, applied by capillary action to the mouthparts (labella) of 15 deer flies (Chrysops sp) and a single horse fly (Tabanus atratus) caused infection in each of 2 sheep. Infection did not occur in 2 calves inoculated daily for 5 days with mouthparts from 50 horn flies collected after feeding on a BLV-infected steer. Four calves receiving bites from 75 stable flies interrupted from feeding on a BLV-positive cow also were not infected. Seronegative cattle held for 1 to 4 months in a screened enclosure with positive cattle in the presence of biting flies were not infected with BLV. The feeding behavior of each insect is discussed to assess their potential as vectors of BLV. PMID- 2982294 TI - A new field strain of equine abortion virus (equine herpesvirus-1) among Kentucky horses. AB - From restriction endonuclease characterization of the DNA of 317 isolates of equine abortion virus (equine herpesvirus-1; EHV-1) from 176 epizootically unrelated outbreaks of equine virus abortion occurring over 24 years in Kentucky, an epizootic pattern and variation of the virus have emerged. Two electropherotypes of EHV-1 (1P and 1B) accounted for greater than 90% of the nonvaccine-related abortion isolates examined. From 1960 to 1981, EHV-1 1P was the predominant isolate circulating in the central Kentucky area and the cause of greater than 80% of EHV-1-related abortions. In 1981, the occurrence of isolate 1B-related abortions began to increase and since 1982, 1B has become the most frequently recovered isolate of EHV-1 from aborted fetuses. PMID- 2982295 TI - Comparison of the pneumopathogenicity of two strains of bovine viral diarrhea virus. AB - The pneumopathogenicity in calves of 2 strains of bovine viral diarrhea (BVD) virus, isolate 2724 (a noncytopathogenic virus) and isolate 72 (a cytopathogenic virus), was compared. All calves were inoculated endobronchially, using fiberoptic bronchoscopy. Two calves were given Pasteurella haemolytica, 2 calves were given the noncytopathogenic BVD virus, and 2 calves were given cytopathogenic BVD virus. Five calves were inoculated sequentially with BVD virus and, 5 days later, with P haemolytica. Two of these calves were inoculated with the noncytopathogenic BVD virus and the other 3 with the cytopathogenic strain. Both BVD virus strains caused marked respiratory tract disease in the calves sequentially inoculated with P haemolytica and also impaired pulmonary clearance of P haemolytica. However, the effect of the cytopathogenic strain was more severe than the noncytopathogenic strain, indicating that strains of BVD virus may vary in their pneumopathogenicity for calves. PMID- 2982296 TI - Equine herpesvirus type 2: cell-virus relationship during persistent cell associated viremia. AB - Some aspects of the biology of equine herpesvirus type 2 (EHV-2) were investigated by examination of the persistent cell-associated viremia stage of the infection. The EHV-2 infection of leukocytes was latent, because free virus was not retrieved without first cultivating harvested leukocytes in vitro. A virus infective center (IC) assay was developed to enumerate latently infected cells in the leukocyte population. This assay proved to be simple and reproducible and revealed a linear relationship between IC plaques formed and the number of cells inoculated, except where large numbers of cells (greater than 4 X 10(6)) were inoculated per 10 cm2 dish. This reduction at high cell densities of IC/10(6) cells inoculated was dependent on cells obtained from an EHV-2-infected horse. There was considerable variation in the numbers of IC/10(6) leukocytes harvested from different horses, but little variation in the harvests from the same horse at different times. There seemed to be a direct relationship between serum-neutralization titers and IC numbers. Transfer of viable infected leukocytes to 2 fetuses failed to establish EHV-2 infection. Infection of equine fetal kidney cells with EHV-2 virus failed to produce detectable Fc receptors on the cell surface. PMID- 2982297 TI - Experimental reproduction of septicemic pasteurellosis in feedlot lambs: bacteriologic and pathologic examinations. AB - Septicemic pasteurellosis (SP) was induced in feedlot lambs. Twenty-eight lambs, randomly allotted into 7 groups, were given combinations of 3 treatments: (i) immunosuppression using hydrocortisone solubilized in dimethyl sulfoxide, (ii) rapid changes in feed, from 100% roughage to 90% concentrate, and (iii) oral inoculation of Pasteurella haemolytica biotype T. Feed changes and immunosuppression by hydrocortisone were needed for the production of SP. Pasteurella haemolytica inoculation was not necessary for induction of SP in all cases, indicating an endogenous source of infection. Clinical pathologic, bacteriologic, and gross and microscopic pathologic findings of induced SP were similar to those described for naturally occurring SP in lambs. Infection of lambs with P haemolytica biotype T via the gastrointestinal tract is discussed as a possible step in the pathogenesis of SP in feedlot lambs. PMID- 2982298 TI - Adenosine triphosphatase activity in the carotid rete-cavernous sinus complex of sheep and goats. AB - The carotid rete-cavernous sinus structures of sheep and goats were frozen with liquid nitrogen or with Freon liquid spray and were cryo-sectioned at -20 C. The main concentration of sodium- and potassium-dependent adenosine triphosphatase was on the tunica intima, especially on the endothelial cells of rete branches and the cavernous sinus. Little reaction product was discernible in the tunica media and the tunica adventitia. PMID- 2982299 TI - Isolation and identification of caliciviruses from dogs with enteric infections. AB - Caliciviruses were isolated from 7 dogs and 1 captured coyote with enteritis. There was a high fatality rate in dogs 4 to 16 weeks of age. The occurrence in these dogs of concurrent infection with known enteric pathogens such as Salmonella sp, canine parvovirus, canine coronavirus, and canine rotavirus did not allow making any conclusions regarding the pathogenicity of this newly recognized calicivirus. The caliciviruses were characterized by electron microscopy and were further identified as being closely related to feline calicivirus by immunoelectron microscopy with specific antibody. PMID- 2982300 TI - Isoproterenol or aminophylline attenuate pulmonary edema after acid lung injury. AB - We tested the hypothesis that isoproterenol and aminophylline would reduce both the pulmonary hypertension and increased pulmonary vascular permeability caused by the intratracheal instillation of hydrochloric acid. The lungs of New Zealand white rabbits were studied using a blood-perfused isolated lung preparation. Changes in pulmonary artery pressure and lung weight were measured for 30 min after acid injury (2 ml/kg 0.1 N HCl). In a control group without acid injury, pulmonary artery pressure and lung weight did not change throughout 1 h of perfusion. In an acid control group, pulmonary artery pressure increased 11 +/- 3 mmHg (p less than 0.01), and lung weight increased 19.2 +/- 5.1 g during the 30 min after acid injury (p less than 0.001). Pretreatment with aminophylline (p less than 0.01) or isoproterenol (p less than 0.001) completely prevented the increase in pulmonary artery pressure after acid instillation. Pretreatment with aminophylline strikingly reduced the increase in lung weight caused by acid injury to 3.6 +/- 1.0 g over 30 min (p less than 0.01). Pretreatment with isoproterenol prevented any increase in lung weight (0.5 +/- 0.5 g over 30 min, p less than 0.01). Post-treatment with isoproterenol beginning 3 min after acid instillation also prevented the increase in pulmonary artery pressure (p less than 0.01) and markedly reduced the increase in lung weight to 5.8 +/- 2.4 g over 30 min (p less than 0.01). The beneficial effect of isoproterenol is mediated through activation of beta-adrenergic receptors because propranolol reverses its effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982301 TI - Antioxidant activity of some antiproteases. AB - Proteolytic enzymes and reactive oxygen molecules participate in the pathogenesis of models of acute inflammation, and antiproteases have been used to determine proteolytic participation in these models. However, antiproteases have been demonstrated to directly inhibit inflammatory cells. In this report, significant antioxidant effects of antiproteases are described. These effects involve both scavenging of oxygen radicals and direct inhibition of enzymes capable of producing oxygen radicals. These results suggest using caution in implicating proteolytic mechanisms solely on the basis of antiprotease inhibition of expected phenomena. PMID- 2982303 TI - Propylthiouracil levels in hyperthyroid patients unresponsive to large doses. Evidence of poor patient compliance. AB - Nine patients with hyperthyroidism due to Graves' disease did not respond to therapy with very large doses (800 to 2000 mg/d) of propylthiouracil. In eight patients, studies showed propylthiouracil was absorbed and metabolized normally. Five patients had no detectable propylthiouracil in their serum 2 to 3 hours after supposedly taking their medication at home, and three patients had markedly abnormal results of perchlorate discharge tests after receiving propylthiouracil under supervision. After evaluation, noncompliance was thought to be the reason for treatment failure in six of the nine patients; one patient was possibly resistant. In two patients, data were insufficient, although intermittent noncompliance could not be ruled out. Among patients who respond poorly to propylthiouracil therapy, noncompliance is the most likely reason. In such patients, methimazole should be substituted for continued massive doses of propylthiouracil. PMID- 2982302 TI - Long-term estrogen replacement therapy prevents bone loss and fractures. AB - Although several case-control studies have shown an inverse association between postmenopausal estrogen use and fractures, quantitation of fracture incidence has been lacking. To quantify the degree to which estrogen replacement therapy prevents postmenopausal osteoporosis, a retrospective study was done comparing the occurrence of fractures in 245 long-term estrogen users and 245 case-matched controls, followed for an average of 17.6 years. Quantitative bone mineral assessments were obtained from 18 women using estrogen replacement therapy and their controls (average age, 73 years). Osteoporotic fracture incidence in estrogen users was 50% as great as in the controls (p less than 0.01). Estrogen users showed significantly greater bone mineral: 54.2% greater spinal mineral (p less than 0.0002), 19.4% greater forearm mineral (p less than 0.0005), and 15.6% greater metacarpal cortical thickness (p less than 0.005). Long-term estrogen replacement therapy confers significant protection against bone loss and fracture. PMID- 2982304 TI - Hypercalcemia and disseminated cytomegalovirus infection in the acquired immunodeficiency syndrome. AB - Two patients with the acquired immunodeficiency syndrome and disseminated cytomegalovirus infection developed hypercalcemia associated with suppressed parathyroid gland activity. Neither patient had evidence of a malignant, endocrinologic, granulomatous, or drug-related cause for hypercalcemia. Increased osteoblastic bone resorption induced by cytomegalovirus infection may have been the cause for the hypercalcemia. Physicians should be alert for the occurrence of endocrinologic and metabolic problems in immunosuppressed patients. PMID- 2982305 TI - Seroepidemiology of human T-lymphotropic virus type III among homosexual men with the acquired immunodeficiency syndrome or generalized lymphadenopathy and among asymptomatic controls in Boston. AB - We studied a cohort of 45 homosexual men with the acquired immunodeficiency syndrome, 78 with persistent unexplained generalized lymphadenopathy, and 160 asymptomatic homosexual controls for serologic evidence of infection with human T lymphotropic virus type III (HTLV-III). Study participants were recruited from a community-based health center and a university hospital practice. Ninety-eight percent of men with the syndrome and greater than 90% of men with generalized lymphadenopathy had antibody to HTLV-III, while 21% of the controls were positive (p less than 0.001). Six patients with generalized lymphadenopathy developed the acquired immunodeficiency syndrome over 2 years; all were seropositive for HTLV III. Thirty-six asymptomatic controls had had sexual contact with a man with the syndrome; receptive anal intercourse in this group was associated with seropositivity for HTLV-III. These data suggest that persistent generalized lymphadenopathy and the acquired immunodeficiency syndrome are part of a clinical spectrum of HTLV-III infection and that most high-risk homosexual men in some regions of the United States have not yet been infected with this virus. PMID- 2982306 TI - Malignant lymphoma presenting as Kaposi's sarcoma in a homosexual man with the acquired immunodeficiency syndrome. AB - A homosexual man had Kaposi's sarcoma of the skin and lymph nodes. After a brief response to interferon, the patient developed new skin lesions. Massive bleeding in the gastrointestinal tract prompted endoscopy, which showed tumor involvement of the stomach, believed to be Kaposi's sarcoma. At autopsy, a diffuse lymphoma was found involving the skin and gastrointestinal tract, forming collision tumors in regions that contained Kaposi's sarcoma. Lymphomatous tissue, but not uninvolved lymph node or spleen, contained Epstein-Barr virus DNA, as shown by DNA hybridization studies. Epstein-Barr virus may play a role in the development of lymphoma in immunosuppressed patients. Unusual manifestations of tumors, such as the massive gastrointestinal bleeding seen in this case, may indicate the development of a second neoplasm. PMID- 2982309 TI - Diagnosis of insulinoma and the euglycemic hyperinsulinemic clamp technique. PMID- 2982307 TI - NIH conference. Clinical applications of corticotropin-releasing factor. AB - Ovine and human corticotropin-releasing factors (CRF) have similar potencies in causing adrenocorticotropic hormone (ACTH) and cortisol secretion in normal humans. Using long-acting ovine CRF (1 microgram/kg body weight as an intravenous bolus), we tested patients with Cushing's syndrome, adrenal insufficiency, and psychiatric conditions with mild hypercortisolism. Over 95% of hypercortisolemic patients with a pituitary adenoma responded with increases in plasma ACTH and cortisol concentrations; patients with the ectopic ACTH syndrome had no ACTH or cortisol responses; patients with ACTH-independent hypercortisolism of adrenal origin had low or undetectable plasma ACTH concentrations before and after CRF without any cortisol response. The differences in responses of patients with adrenal insufficiency of primary, pituitary, or suprapituitary type likewise suggest value of the CFR test in their differential diagnosis. The responses in the psychiatric patients should permit differentiation between Cushing's syndrome and hypercortisolism of psychiatric origin. PMID- 2982308 TI - Thyroid hormone action at the nuclear level. AB - Recent findings have led to a greater understanding of thyroid hormone action. The nuclear receptor for triiodothyronine is an integral component of a larger chromatin fragment. A stereospecific energy-dependent transport system appears responsible for translocation of triiodothyronine from cytosol to nucleus. In the liver, a multiplicative interaction between a signal from the triiodothyronine nuclear receptor complex and a signal generated from carbohydrate metabolism results in the induction of specific mRNAs. Two-dimensional mRNA activity profiles suggest that approximately 8% of the visible mRNA sequences are differentially affected by alterations in thyroid states. Almost 30% to 40% of these changes are mediated by an increase in pituitary growth hormone induced by triiodothyronine. Sequential analyses of mRNA activity profiles have identified an mRNA sequence (mRNAs14) coding for a protein (S14) with Mr 17 010 and pI 4.9 which responds to triiodothyronine with a lag time of less than 20 minutes. The coordinate regulation of mRNAs14 by carbohydrate and triiodothyronine and its presence in lipogenic tissues (fat, liver, lactating mammary tissue) suggests that S14 is involved in some aspect of fatty acid synthesis degradation or storage. PMID- 2982310 TI - Effect of propranolol on peroxidase in tracheal epithelial cells. AB - Propranolol (beta-blocker) was administered to five guinea pigs in doses of 6 mg/day for ten days; five uninjected animals were used as controls. The first or second tracheal ring was cut in round slices. The number of overall epithelial cells facing the airway and the epithelial cells with peroxidase were counted. The mean percentage of peroxidase-positive cells to total epithelial cells was 49% in the control and 11% in animals after the administration of propranolol. This result apparently meant that the peroxidase activity was decreased by the administration of propranolol. The relationship between allergic diseases and the synthesis of peroxidase is discussed. PMID- 2982311 TI - Glycosaminoglycan in the blood and renal tissue of a patient with nephroblastomatosis. AB - Bilateral nephroblastomatosis was diagnosed in a 15-month-old white female. Prior to surgery, multiple peripheral blood smears (Wrights' stain) revealed an azurophilic staining extracellular material. When serum was added to a three percent acetic acid solution, a floccular, fibrous precipitate formed at the meniscus of the tube. Serum protein electrophoresis on cellulose acetate support media resulted in a distorted pattern which corrected to a normal pattern upon treatment with hyaluronidase. These peripheral blood abnormalities disappeared following a left nephrectomy. Quantitative chemical analysis of diseased renal tissue yielded 81 micrograms of readily extracted glycosaminoglycan (GAG) per gram of tissue. The importance of abnormal glycosaminoglycan production in patients with malignant disease is discussed both in terms of clinical importance and possible roles of cell exudates. PMID- 2982312 TI - Potential implication of genetic engineering and other biotechnologies to insect control. PMID- 2982314 TI - A comparative study of the resonance Raman spectra of bacterial cytochromes. AB - Resonance Raman spectra were obtained for two newly isolated bacterial cytochromes, Alcaligenes faecalis (ATCC 8750) c554 and Alcaligenes faecalis c556. Their spectra were compared with those of mammalian cytochrome c and two other bacterial cytochromes, Paracoccus denitrificans c550 and Pseudomonas aeruginosa c551. The positions of the Raman bands indicated that, while Al. c554 and Al. c556 were c-type cytochromes with two thioether linkages, several common features found in their Raman spectra were anomalous. These features suggest that the electronic charge density of the porphyrin macrocycle of Al. c554 and Al. c556 is more asymmetric than that of other bacterial and mammalian c-type cytochromes. The Raman evidence indicates that the electronic properties of the heme are controlled by the protein in these two Alcaligenes cytochromes. PMID- 2982313 TI - The membrane-bound alkaline phosphatase and 5'-nucleotidase activities of vegetative cells of Dictyostelium discoideum. AB - Earlier reports suggested that the adenosine monophosphate (AMP)- and the p nitrophenyl phosphate (pNPP)-hydrolyzing activities of Dictyostelium discoideum membrane preparations are due to different proteins. These results have been apparently contradicted by the recent purification to homogeneity of the two activities from culmination phase cells as a single protein [D. R. Armant and C. L. Rutherford (1981) J. Biol. Chem. 256, 12710-12718]. Results presented here from studies on the activities of vegetative cells support the concept of a single protein. Nondenaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of Triton X-100 extracts of cell membrane preparations of D. discoideum showed identical migration of pNPPase and AMPase activities. Furthermore, the previously reported different pH optima of the two activities was due to the fact that pH optima are dependent upon the substrate concentration, and the selective solubilization of AMPase from membrane preparations by phospholipase C can probably be accounted for by the finding that phospholipase C preparations from the same commercial source contain 5' nucleotidase activity. Moreover, there are alterations in the Km and the stability of both AMPase and pNPPase in a strain with a mutationally altered alkaline phosphatase, further supporting the concept that the two activities are due to a single protein. Both substrates serve as transphosphorylation donors demonstrating that the enzyme activity is mechanistically an alkaline phosphatase. PMID- 2982316 TI - Thyroid hormone synthesis and thyroglobulin iodination related to the peroxidase localization of oxidizing equivalents: studies with cytochrome c peroxidase and horseradish peroxidase. AB - Cytochrome c peroxidase (CcP) and horseradish peroxidase (HRP), when combined with a stoichiometric amount of H2O2, form stable compounds I which are known as FeIV Ro and FeIV o pi + structures, respectively. These compounds were assayed in the catalysis of thyroid hormone synthesis and the iodination reaction. As previously shown for the lactoperoxidase FeIV Ro compound, the CcP FeIV Ro compound was involved in the coupling and not in the iodination reactions. In contrast, the HRP FeIV o pi + compound catalyzed both iodination and hormone formation. The possible role of the different peroxidase-H2O2 compounds in the two sequential reactions, thyroglobulin iodination and thyroid hormone formation, is discussed. PMID- 2982315 TI - 10-Keto or 25-hydroxy substitution confer equivalent in vitro bone-resorbing activity to vitamin D3. AB - The biological activities of 10-keto derivatives of vitamin D3 and 25 hydroxyvitamin D3 were determined in bone organ culture. Fetal rat limb bones prelabeled with 45Ca were incubated for 60 h with 10-keto-25-hydroxyvitamin D3, 10-keto-vitamin D3, 1,25-dihydroxyvitamin D3, 25-hydroxyvitamin D3, or vitamin D3. Resorption was quantified by release of 45Ca. Substitution of a keto group in the 10 position of the vitamin D3 molecule resulted in a compound equal in potency to 25-hydroxyvitamin D3. When a 10-keto group was substituted in the 25 hydroxy vitamin D3 molecule, the potency was increased 20- to 40-fold. In contrast, 1,25-dihydroxyvitamin D3 was 7500-fold more potent than 25 hydroxyvitamin D3. Since 10-keto-25-hydroxyvitamin D3 has a retention time close to that of 1,25-dihydroxyvitamin D3 on normal-phase HPLC eluted with isopropanol:hexane, it is a possible artifact in the assay of 1,25 dihydroxyvitamin D3. Based upon the observed relative activities of the two compounds, the concentration of 10-keto-25-hydroxyvitamin D3 would have to be greater than 0.8 ng/ml for it to interfere in the bioassay of 1,25 dihydroxyvitamin D3. PMID- 2982317 TI - Galactose-rich glycoproteins are on the cell surface of herpes virus-infected cells. 1. Surface labeling and serial lectin binding studies of Asn-linked oligosaccharides of glycoprotein gC. AB - Cell-surface glycoproteins of mock-infected and herpes simplex virus type 1 (HSV 1)-infected BHK-21 and HEp-2 cells were radiolabeled by incubation with galactose oxidase followed by reduction with NaB3H4. The incorporation of radiolabel into glycoconjugates in both BHK-21 and HEp-2 cells was increased several fold following infection with HSV, showing an increase in surface-exposed Gal residues in the infected cells. This was further confirmed by an increase in binding of cell-surface-labeled glycoproteins gC and gB from HSV-infected BHK-21 cells to Ricinus communis agglutinin I, which is specific for beta-D-Gal residues. Prior treatment of cells with Clostridium perfringens neuraminidase enhanced the surface radiolabeling by the galactose oxidase/NaB3H4 method: HEp-2 cells exhibited over sixfold enhancement in labeling, while BHK-21 cells showed only a slight increase. HSV glycoprotein gC was the predominant cell-surface glycoprotein radiolabeled by the galactose oxidase/NaB3H4 method in virus infected BHK-21 cells. The glycoprotein gC was purified by immunoaffinity column chromatography on monoclonal anti-gC-antibody-Sepharose. The radiolabel in the glycopeptides of gC was resistant to beta elimination, showing that it was associated only with Asn-linked oligosaccharides. A serial lectin affinity chromatography of glycopeptides on columns of concanavalin A-Sepharose, lentil (Lens culinaris) lectin-Sepharose, and Ricin I-agarose allowed the assignment of minimal oligosaccharide structures bearing terminal Gal residues in gC. PMID- 2982318 TI - A monoclonal antibody to the epsilon-aminocaproic acid binding site on the kringle 4 region of human plasminogen that accelerates the activation of Glu1 plasminogen by urokinase. AB - A monoclonal antibody, 10-F-1, previously shown [V. A. Ploplis, H. S. Cummings, and F. J. Castellino (1982) Biochemistry 21, 5891-5897] to interact with a particular epsilon-aminocaproic acid (EACA)3 binding site on the kringle 4 (K4) region of human Glu1-plasminogen (Glu1-Pg), has been employed to assess the contribution of this particular EACA site toward the enhancement, by EACA and its analogs, of the urokinase (UK)-catalyzed activation of Glu1-Pg. As is the case with EACA-like compounds, the presence of antibody 10-F-1 accelerates the activation of Glu1-Pg by UK, but does not enhance the similar activation of Lys77 plasminogen. In the presence of concentrations of antibody 10-F-1 which saturate its binding site on Glu1-Pg, the Km of Glu1-Pg activation by UK is raised from 1.4 +/- 0.2 microM, a value obtained in the absence of antibody, to 17.0 +/- 2.0 microM. On the other hand, the kcat for this activation, 0.038 +/- 0.005 s-1, is elevated to 2.45 +/- 0.2 s-1 at saturating concentrations of antibody 10-F-1. The kcat/Km for activation under these conditions is 0.027 s-1 microM-1 in the absence of antibody, and 0.144 s-1 microM-1 in the presence of saturating levels of antibody 10-F-1. This demonstrates that the interaction of this antibody with its epitope results in a fivefold stimulation of the activation rate of Glu1-Pg by UK. The availability of antibody 10-F-1 allows for a specific means of probing the function of one of the four to five thermodynamically equivalent weak EACA sites on human plasminogen. From this particular study, it is concluded that the weak binding site for EACA on the K4 domain of Glu1-Pg is either in-part or in whole responsible for the enhancing effect of EACA on human Glu1-Pg activation by UK. PMID- 2982319 TI - Molecular conversion between monomeric and dimeric states of the mitochondrial cytochrome b-c1 complex: isolation of active monomers. AB - Bovine heart cytochrome b-c1 complex dispersed in 0.1% dodecylmaltoside, 10 mM Tris-HCl (pH 7.4), was subjected to filtration on Ultrogel AcA 34 columns. Apparent Mr values of about 400,000 and 170,000 were estimated for the enzyme detergent complex in the presence and absence of 50 mM KCl, respectively. Similar Mr values (about 390,000 and 160,000) were obtained after sucrose gradient centrifugation of the b-c1 complex species isolated using Ultrogel filtration. Both species contained eight polypeptides, as in the original cytochrome b-c1 complex. The experiments suggest that the two species represent a dimer and a monomer of the b-c1 complex. The molecular conversion between the monomeric and dimeric state of the enzyme was found to be reversible. Both monomers and dimers of the b-c1 complex were competent to catalyze QH2:cytochrome c reductase activity with approximately the same maximal velocity. The finding that both molecular forms of the enzyme appear equally active does not support functional models based exclusively on a dimeric b-c1 complex. PMID- 2982320 TI - Structural alterations of the inner mitochondrial membrane in ischemic liver cell injury. AB - Mitoplasts were prepared from 3-h ischemic livers in an attempt to define the structural alterations in the inner membrane that may account for the functional deficiencies of ischemic mitochondria. Mitoplasts from both control and ischemic livers had similar specific activities of cytochrome oxidase and succinate cytochrome c reductase. With both preparations, the specific activity of rotenone insensitive NADH-cytochrome c reductase was 10-fold lower than in the mitochondria from which they were prepared. Ischemic mitoplasts had no respiratory control with ADP, and had a slightly reduced phospholipid to protein ratio and an increased cholesterol to protein ratio. As a result, the cholesterol to phospholipid molar ratio was increased from the control of 0.04 to 0.08. There were also differences in the content of individual phospholipid species. Phosphatidylcholine increased by 15%, while cardiolipin decreased by 60%. There were increases in sphingomyelin and in the lysophospholipids of phosphatidylcholine, ethanolamine, and cardiolipin. Pretreatment with chlorpromazine did not prevent these changes. Linoleic acid was decreased by 35% in ischemic phospholipids, and the content of free fatty acids was increased 4 fold. Electron spin resonance spectroscopy of mitoplasts spin labeled with either 5- or 12-doxyl stearic acid revealed an increased molecular order (decreased fluidity) of ischemic inner mitochondrial membranes consistent with the increased cholesterol to phospholipid ratio. The data indicate activation of a phospholipase A in ischemic mitochondria with the resulting accumulation of products of lipid hydrolysis. This conclusion further emphasizes the close similarity between the structural and functional consequences of ischemia in the intact animal and the effect on isolated mitochondria of the activation of the endogenous phospholipase A. In both cases the major functional alterations are attributable to changes in the permeability of the inner mitochondrial membrane induced by the accumulation of lysophospholipids. PMID- 2982321 TI - Evidence for a reactive cysteine at the nucleotide binding site of spinach ribulose-5-phosphate kinase. AB - Ribulose-5-phosphate kinase from spinach was rapidly inactivated by N bromoacetylethanolamine phosphate in a bimolecular fashion with a k2 of 2.0 M-1 S 1 at 2 degrees C and pH 8.0. Ribulose 5-phosphate had little effect on the rate of inactivation, whereas complete protection was afforded by ADP or ATP. The extent of incorporation as determined with 14C-labeled reagent was about 1 molar equivalent per subunit in the presence of ATP with full retention of enzymatic activity, and about 2 molar equivalents per subunit in the completely inactivated enzyme. Amino acid analyses of enzyme derivatized with 14C-labeled reagent reveal that all of the covalently incorporated reagent was associated with cysteinyl residues. Hence two sulfhydryls are reactive, but the inactivation correlates with alkylation of one cysteinyl residue at or near the enzyme's nucleotide binding site. The kinase was also extremely sensitive to the sulfhydryl reagents 5,5'-dithiobis(2-nitrobenzoic acid) and N-ethyl-maleimide. The reactive sulfhydryl groups are likely those generated by reduction of a disulfide during activation. PMID- 2982322 TI - Biochemical differences between subsarcolemmal and interfibrillar mitochondria from rat cardiac muscle: effects of procedural manipulations. AB - Differences in oxidative metabolism between subsarcolemmal and interfibrillar heart mitochondria were investigated. Interfibrillar mitochondria oxidized substrates donating reducing equivalents at Complex I (NADH-CoQ reductase), Complex II (succinate-CoQ reductase), and Complex III (CoQH2-cytochrome c reductase) more rapidly than did subsarcolemmal mitochondria. There was no difference in oxidation of substrates entering the electron transport chain at Complex IV (cytochrome c oxidase). Differences expressed in normal-ionic-strength medium at Complexes II and III but not I were eliminated in low-ionic-strength medium. The concentrations of cytochromes and activities of NADH and cytochrome c oxidase were virtually the same in the two populations. In permeabilized mitochondria, activities of succinate-duroquinone and TMPD plus ascorbate oxidase were significantly lower in the subsarcolemmal mitochondria. Differences in membrane permeability between the populations were suggested by the greater permeability of subsarcolemmal mitochondria to exogenous NADH. The influence of isolation buffers and preparative procedures on the two classes of mitochondria were also examined. Characteristic biochemical and morphological properties of the two populations were unchanged by exposing each to the preparative procedure used to isolate the alternate population; the oxidative performance of the two populations cannot be equalized by experimental manipulation. PMID- 2982323 TI - A rapid-scan spectrometric and stopped-flow study of compound I and compound II of Pseudomonas cytochrome c peroxidase. AB - A quantitative yield of half-reduced (ferrous-ferric) cytochrome c peroxidase from Pseudomonas aeruginosa has been obtained by using either ascorbate or NADH as reductant of the resting (ferric-ferric) enzyme along with phenazine methosulfate as mediator. The formation of Compounds I and II from the half reduced enzyme and hydrogen peroxide has been studied at 25 degrees C using rapid scan spectrometry and stopped-flow measurements. The spectra of Compound I in the Soret and visible regions were recorded within 5 ms after mixing the half-reduced enzyme with H2O2. The spectrum of the primary compound at the Soret region had a maximum at 414 nm, and in the visible region at 528 and 556 nm. The spectrum of Compound I showed no bands in the 650-nm region, excluding the possibility of a pi-cation radical being part of the catalytic mechanism. Compound I was stable for at least 12 s when no reducing equivalents were present. In the presence of reduced azurin, half-reduced enzyme reacted with H2O2 to form Compound II within 50 ms. The spectrum of Compound II had a Soret maximum at 411 nm. In the visible region the Compound II spectrum was close to that of the totally oxidized, resting enzyme form. In the presence of excess azurin, Compound II was converted rapidly to the half-reduced enzyme form. The kinetics of Compound I formation was also followed with peracetic acid, ethylhydroperoxide, and m-chloroperbenzoic acid as electron acceptors. The rate constants of these reactions are diminished compared to that of hydrogen peroxide, indicating a closed structure for the heme pocket of the enzyme. PMID- 2982324 TI - The phosphate-pyrophosphate exchange and hydrolytic reactions of the membrane bound pyrophosphatase of Rhodospirillum rubrum: effects of Mg2+, phosphate, and pyrophosphate. AB - The relation that exists between the Pi-PPi exchange reaction and pyrophosphate hydrolysis by the membrane-bound pyrophosphatase of chromatophores of Rhodospirillum rubrum was studied. The two reactions have a markedly different requirement for added Mg2+. Optimal rates of hydrolysis were attained at 1 mM Mg2+ with 0.67 mM pyrophosphate; the rate od hydrolysis correlated with the concentration of Mg-pyrophosphate, which indicated that the latter was the substrate for hydrolysis. The Pi-PPi exchange reaction rate was low at concentrations of added Mg2+ below 1 mM (0.67 mM pyrophosphate), but increased as the concentration of Mg2+ in the medium was increased. The Pi-PPi exchange reaction depends on the concentration of MgHPO4, which suggests that this is the substrate in the exchange reaction. However, it is likely that free Mg2+ also exerts a favorable effect on the Pi-PPi exchange reaction. The optimal concentration for the Pi-PPI exchange reaction was approx 240 microM, which suggests that the concentration of the hydrolyzable substrates modulates the kinetic characteristics of the enzyme. PMID- 2982325 TI - Binding and function of mitochondrial glycerol kinase in comparison with those of mitochondrial hexokinase. AB - Mitochondrial-bound glycerol kinase in rat brain was examined with reference to factors involved in the binding and significance of the binding in relation to ATP metabolism inside the mitochondria. The mitochondrial-bound glycerol kinase was solubilized with glycerol 3-phosphate or ADP, and the solubilized enzyme was rebound to mitochondria by addition of divalent cations. The rebinding was decreased by the presence of glycerol 3-phosphate, while was increased by glucose 6-phosphate. Positive correlation was found between the formation of glycerol 3 phosphate by mitochondrial-bound glycerol kinase and ATP content in mitochondria in experiments using various concentrations of succinate and ADP. On the other hand, glycerol 3-phosphate formation was inhibited by addition of inhibitors for mitochondria functions, such as oligomycin, dinitrophenol, cyanide, and atractyloside. Furthermore, formation of dihydroxyacetone phosphate from glycerol was proved, indicating the involvement of glycerol kinase in glycerol phosphate shuttle in combination with glycerol phosphate dehydrogenase. These findings are discussed in comparison with those of mitochondrial-bound hexokinase. PMID- 2982326 TI - Insulin mediator causes dephosphorylation of the alpha subunit of pyruvate dehydrogenase by stimulating phosphatase activity. AB - Insulin treatment of rats results in an increased amount or activity of insulin mediators in liver and skeletal muscle. These mediators stimulated pyruvate dehydrogenase and inhibited adenylate cyclase. The insulin-generated mediators caused dephosphorylation of the alpha subunit of pyruvate dehydrogenase in mitochondria prelabeled with [gamma-32P]ATP. An assay was developed which quantitatively measured mediator activity by determining the rate of alpha subunit dephosphorylation. The dephosphorylation was directly proportional to the amount of mediator added and was directly related to activation of pyruvate dehydrogenase. The decrease of alpha-subunit phosphorylation resulted from stimulation of pyruvate dehydrogenase phosphatase, since it occurred in the absence of ATP and was inhibited by NaF. These data further delineate the mechanism of insulin mediator activation of pyruvate dehydrogenase. PMID- 2982327 TI - The reactions of the phosphorylated pathway of L-serine biosynthesis: thermodynamic relationships in rabbit liver in vivo. AB - The thermodynamic relationships among the reactions of the phosphorylated pathway of L-serine biosynthesis have been determined in rabbit liver in vivo in different dietary states. The mass action ratios of the reactions involved were calculated from the concentrations of appropriate metabolites in freeze-clamped liver and compared with the equilibrium constants of the same reactions previously determined under physiological conditions. Toward this goal, a new, highly specific enzymatic assay for L-phosphoserine was developed to allow the accurate measurement of this intermediate in biological material. The level of L phosphoserine, the immediate precursor of L-serine, varied significantly with diet, being 0.81, 0.38, and 0.21 mumol/g wet wt in the fed, and 24 h and 48 h fasted states, respectively. The tissue content of L-phosphoserine was also sensitive to anoxia, falling almost fivefold within 5 min after the liver was removed. Values of for the combined reactions of the first two steps of the pathway of L-serine biosynthesis [D-3-phosphoglycerate dehydrogenase (EC 1.1.1.95) and L-phosphoserine aminotransferase (EC 2.6.1.52)] in livers from animals in different dietary states were calculated to be 1.2 X 10(-4) (fed), 1.4 X 10(-4) (24 h starved), and 0.70 X 10(-4) (48 h starved), all being very close to the value of the combined equilibrium constant of the same reactions (2.44 X 10(-4). Even when there were major changes in the individual components of, such as a fivefold drop in L-phosphoserine and a sevenfold fall in alpha-ketoglutarate following 5 min of anoxia, remained relatively unchanged (2.7 X 10(-4). Thus, it has been concluded that, in rabbit liver under most normal conditions, the combined reactions of D-3-phosphoglycerate dehydrogenase and L-phosphoserine aminotransferase remain very near equilibrium, and that almost all of the disequilibrium of the pathway, amounting to a delta G of -5.5 kcal/mol in the fed state, is at the last step, the L-phosphoserine phosphatase reaction (EC 3.1.3.3). PMID- 2982328 TI - Thyroid hormone regulation of flavocoenzyme biosynthesis. AB - The means by which thyroid hormone regulates flavocoenzyme biosynthesis was studied in hyper-, eu-, and hypothyroid rats by determining the activities of flavocoenzyme-forming enzymes, viz., flavokinase and FAD synthetase, as well as those of flavocoenzyme-degrading enzymes, viz., FMN phosphatase and FAD pyrophosphatase. Flavokinase activity was increased in hyperthyroid animal and decreased in hypothyroid animals. Correspondence of flavokinase activity with the amount of a high-affinity flavin-binding protein quantitated immunologically in hypo-, eu-, and hyperthyroid rats indicated that the thyroid response is caused by an increased amount of enzyme; moreover, the concomitant decrease in a low affinity flavin-binding protein suggests an inactive precursor form of flavokinase. FAD synthetase activity showed a similar but less pronounced trend than flavokinase. Activities of FMN phosphatase and FAD pyrophosphatase were not influenced by thyroid hormone. Overall results indicate that the mechanism of thyroid hormone regulation of flavocoenzyme level is in the steps of biosynthesis, especially at flavokinase, rather than in degradation steps. PMID- 2982329 TI - Deglycosylated mammalian beta 2-adrenergic receptors: effect on radioligand binding and peptide mapping. AB - Mammalian beta-adrenergic receptors are glycoproteins containing both high mannose and complex-type carbohydrate chains [G. L. Stiles, J. L. Benovic, M. G. Caron, and R. J. Lefkowitz (1984) J. Biol. Chem. 259, 8655-8663]. Endoglycosidase F treatment of beta 2-adrenergic receptors results in the removal of at least two N-linked oligosaccharides, resulting in an increased mobility of the receptor peptide on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Mr 62,000 to 49,000). In the present study the properties of the deglycosylated beta 2 adrenergic receptor were assessed. Following deglycosylation, the beta 2 adrenergic receptor recognized both agonists and antagonists with the same potency order and affinities as the glycosylated form. In addition, two dimensional gel electrophoresis peptide mapping techniques applied to control and deglycosylated beta 2-adrenergic receptors (both within and between species) demonstrated that there was a marked homology of the beta 2-adrenergic receptor between species which are closely related phylogenetically. In addition, the glycan component of the receptor did not appear to interfere with the ability of proteinases to generate accurate peptide maps. PMID- 2982330 TI - Calmodulin-dependent NAD kinase of human neutrophils. AB - NAD kinase from human neutrophils has been partially purified by sequential application of Red Agarose, ion-exchange, and gel-filtration chromatography. The enzyme has a broad pH optimum, 7.0-9.5, is strictly dependent upon the presence of Mg2+, and in the absence of calcium exhibits Km values of 0.6 and 0.9 mM for NAD and ATP, respectively. NAD kinase activity is extremely sensitive to free calcium concentration, with half-maximal activity observed at free calcium concentrations of approximately 0.4 microM. In cellular extracts calcium dependent activation of NAD kinase increases the maximum velocity of the reaction from 2- to 5-fold while not affecting Km values for NAD and ATP. The activity of the partially purified NAD kinase is stimulated 3.5-fold by the addition of calmodulin in the presence of calcium. This stimulation is inhibited by the addition of 20 microM trifluoperazine to the incubation. These data are interpreted as implicating calmodulin in NAD kinase regulation. The total concentration of NADP + NADPH in the human neutrophil used increased 2.2-fold in response to activation by phorbol myristic acetate. Finally, neutrophil NAD kinase has a Mr, based upon gel filtration, of 169,000. PMID- 2982331 TI - [Radiotherapy of metastatic bone tumor with a synthetic calcitonin derivative (Elcatonin) with irradiation]. AB - Twenty cases of metastatic bone tumors were treated with Elcatonin together with irradiation, achieving pain relief in 47.3% of cases, improved bone scintigrams in 16.7% of cases and radiographic improvement of invaded bone in 7.7% of cases. It has reported that calcitonin is effective for pain relief in 50% of cases but for the radiographic improvement of bone lesions in only 0-10%. The life quality of patients with bone metastases is controversial since treatment techniques for advanced oncological patients has progressed remarkably. The pain relief obtained with calcitonin contributes significantly to improving the quality of life for such patients when administered in combination with conventional treatment modes. PMID- 2982332 TI - [Phase II study of VP-16 (capsule) in solid tumors. A cooperative study]. AB - A phase II study of VP-16, a semisynthetic Podophyllotoxin, was performed in patients with solid tumors. VP-16 was administered orally at a dose of 200mg/day for 5 consecutive days at 3 to 4-week intervals. Out of 41 patients who were entered into the study, 35 patients comprising 17 lung cancer, 10 hepatoma and 8 other tumors were evaluable. There were 4 partial responses (23.5%) for lung cancer, 1 (10.0%) for hepatoma and 1 for rhabdomyosarcoma. Overall response rate was 18.2% for patients with prior chemotherapy and 15.4% for those given no prior chemotherapy respectively. Thus the results indicated VP-16 has no cross resistance to other antitumor agents. Leukopenia (less than 4,000/mm3) and thrombocytopenia (less than 10 X 10(4)/mm3) were observed in 72.7% and 29.4% of the patients, respectively. Other toxicities were alopecia (59.5%) and gastrointestinal disturbances such as nausea (46.2%), vomiting (20.5%) and anorexia (20.5%), but these were all well tolerated. PMID- 2982333 TI - [Enhanced cytotoxic effect of Adriamycin and vincristine against PC-6 (human lung cancer cell line) and Hattori (human breast cancer cell line)]. AB - Enhancements of cytotoxic effects of adriamycin (ADM) and vincristine (VCR) on PC 6 (lung cancer cell line) and Hattori (breast cancer cell line) were investigated by concomitant use of a calmodulin inhibitor: nicardipine, antiplatelet agents: oxyfedrine and trimethazidine and an antihypertensive agent: trichlormethiazide. Nicardipine increased the cytotoxic activity of both drugs but other agents did not produce any significant increase. Nicardipine increased the cytocidal effect of VCR on PC-6 about 2-fold and on Hattori about 3-fold. Furthermore, it also increased the effect of ADM on Hattori about 2-fold. Thus, nicardipine enhanced the cytocidal activity of VCR more remarkably than that of ADM. PMID- 2982334 TI - Factors predisposing to abnormal pulmonary function after adenovirus type 7 pneumonia. PMID- 2982335 TI - Serum angiotensin-converting enzyme in sarcoidosis and psoriasis. AB - Untreated pulmonary sarcoidosis is associated with an increased level of serum angiotensin-converting enzyme (SACE), which is regarded as a valuable method of diagnosing sarcoidosis and measuring the activity of the disease. The level of SACE in cutaneous sarcoidosis or other skin diseases has not been clearly established. We therefore examined SACE in 31 patients with systemic sarcoidosis, including cutaneous manifestations, and 12 patients with isolated cutaneous sarcoidosis. Also, 23 patients with psoriasis were studied. The level of SACE was generally elevated only in patients with untreated systemic sarcoidosis, whereas it was normal in cutaneous sarcoidosis and psoriasis. If the level of SACE is elevated in "isolated" cutaneous sarcoidosis, systemic disease must be strongly suspected. PMID- 2982336 TI - Phagocytosis of brushite crystals by pig neutrophils. AB - Neutrophils from pig blood were disrupted by homogenisation or sonication and placed on an analytical sucrose gradient. Pig neutrophil azurophil granules were less dense than the specific granules, unlike those from neutrophils of most other mammalian species. Brushite crystals, which stimulate the respiratory burst in pig neutrophils, were found by electron microscopy to be phagocytosed. Membrane-limited vesicles containing crystals were obtained from a dense region of the sucrose gradient loaded with a homogenate of crystal treated cells. Intake of crystals involved preferential mobilisation of cytochrome b from the plasma membrane and the fusion of both specific and azurophil granules with the primary phagosome. Plasma membrane and granule markers appear in a crystal-containing region of the sucrose gradient when cells are treated with crystals. They are present in much lower concentration at this location in gradients from cells untreated with a crystal stimulus. PMID- 2982337 TI - The use of pathologic features in selecting the extent of surgical resection necessary for breast cancer patients treated by primary radiation therapy. AB - The extent of the surgical resection necessary for breast cancer patients treated by primary radiation therapy is unknown. A simple gross excision of the tumor provides the best cosmetic result, but a wide local resection may be important to prevent local recurrence in some patients. In order to identify patients who are not adequately treated by gross excision of the tumor and radiation therapy, we performed a retrospective clinical-pathologic review of 221 treated women with infiltrating duct carcinoma. There were 53 cases in which the excision specimen showed a constellation of three pathologic features: prominent intraductal carcinoma in the tumor, intraductal carcinoma in the grossly-normal adjacent tissue, and poorly-differentiated nuclei. These cases had a 37% risk of a local recurrence at 6 years compared to eight per cent for all other cases (p less than 0.0001). In cases with all three features, the use of a supplemental dose of radiation to the primary site did not significantly reduce the risk of a local recurrence. Local recurrence at 6 years was 34% in cases with all three features, who received supplemental local radiation, compared to 49% in cases not receiving a supplemental dose (p = 0.28). Survival was also worse for patients with all three features compared to other cases (69% vs. 90% at 6 years, p = 0.002). These results indicate that patients with all three pathologic features have a high risk of local recurrence following gross excision of the tumor and radiation therapy. If primary radiation therapy is selected for these patients, they should first undergo a re-excision of the tumor site in order to be certain that areas of extensive intraductal carcinoma have been adequately resected. Patients whose tumors do not show all three features are adequately treated by gross excision of the tumor prior to radiation therapy. PMID- 2982338 TI - Vulvar sweat gland carcinomas. AB - Sweat gland carcinomas constitute less than 10% of all malignant tumors of the vulva, including those associated with extramammary Paget's disease (EPD). The histopathologic diagnosis of vulvar sweat gland carcinomas is difficult because of their rarity; their resemblance to metastatic carcinomas of the internal female genitalia, kidneys, and other sites; and their diversity of microscopic appearances. We report five examples of vulvar sweat gland carcinomas, two of which were associated with EPD. The other tumors included one example each of ductal eccrine adenocarcinoma, eccrine porocarcinoma, and clear cell hidradenocarcinoma. The clinical behavior of these neoplasms is correlated with their histologic types; we also discuss differential diagnosis and treatment recommendations. PMID- 2982339 TI - Neuron-specific enolase immunoreactivity in small-cell carcinoma of the uterine cervix. PMID- 2982340 TI - Wilms' tumor and glomerulopathy. PMID- 2982341 TI - Acute inflammation in malignant fibrous histiocytoma. PMID- 2982342 TI - Fabry's disease in a heterozygous woman. AB - Fabry's disease is a rare, X-linked disorder of glycolipid metabolism that is usually recognized in men in the third and fourth decades of life. A heterozygous state in women has been documented, but reports about women who are affected are confined to isolated case reports and to rare small series. We report a case of a 42-year-old woman with persistent proteinuria who was found to have Fabry's disease. The characteristic lesion was readily seen using routine high-resolution light microscopy (HRLM) tissue sections without the need of multiple extra tissue sections and special stains. We discuss the correlation of clinical findings, fluorescent microscopy, HRLM, electron microscopy, and biochemical enzymatic studies in establishing the diagnosis. PMID- 2982343 TI - Arterial regenerative activity after prosthetic implantation. AB - Factors modifying arterial regenerative activity were evaluated by replacing adult rabbit aortas with either absorbable polyglycolic acid (PGA) or nonabsorbable Dacron prostheses, 3.5 mm in internal diameter by 24 mm in length, woven to identical specifications including pore size, wall thickness, and elastic modulus, and were followed up for 12 months. At death, 48 PGA and 20 Dacron specimens were studied grossly and by arteriography, light microscopy, scanning and transmission electron microscopy, and by bursting strength determinations. There were no aortic-related deaths or transaortic hemorrhages. Both materials elicited a surrounding inflammatory reaction containing macrophages which transgressed the interstices of only the PGA prostheses. Between two and four weeks, circumferentially oriented smooth-musclelike myofibroblasts proliferated in the PGA inner capsule yielding a neointima 3.2 times thicker than Dacron's. Early ultrastructurally primitive cells progressively differentiated into these smooth-musclelike myofibroblasts. The few myofibroblasts later appearing with Dacron were radially oriented. After one month proliferation stopped in both groups and neointimal thickness became constant. The luminal surface in the PGA group was endothelial-like but was fibrinous in the Dacron group. All specimens withstood saline infusion at three to five times systolic pressure. These studies demonstrate great arterial regenerative potential and suggest hemodynamic and intercellular mitogens controlling it. PMID- 2982344 TI - Experimental myocardial ischemia. Differential injury of mitochondrial subpopulations. AB - Distinct populations of subsarcolemmal (SS) and inter-fibrillar (IF) rat cardiac mitochondria were studied following 15 and 30 minutes of warm and cold global ischemia. The respiratory control index, state 3, state 4, adenosine diphosphate oxygen ratio, and specific enzyme activities of these mitochondrial populations were examined. The subsarcolemmar and IF mitochondria were both severely uncoupled and inhibited by warm ischemia. However, IF mitochondria had a higher RCI at each ischemic interval. In cold ischemia, IF mitochondria were not injured compared with control specimens. Subsarcolemmar mitochondria showed a trend towards a lower RCI that was statistically significant at 30 minutes with succinate as a substrate. These data implicate a differential injury of ischemia on the compartmentalized bioenergy metabolism of the myocardial cell. PMID- 2982345 TI - Poliovirus neutralization and the percentage law. Brief report. AB - The degree of neutralization of poliovirus by mono- and polyclonal antibodies is determined by the antibody/virion ratio at virus concentrations exceeding 10(8) PFU/ml, and by the antibody concentration alone at lower virus doses. PMID- 2982346 TI - Isolation of a new serotype of bovine rhinovirus from cattle. Brief report. AB - Six strains of a new serological type of bovine rhinovirus were isolated in primary calf kidney cell cultures from nasal swab specimens of cattle with acute respiratory disease in Japan. They are serologically indistinguishable from one another and show no cross reaction with bovine rhinovirus types 1 and 2. We propose that the new isolates classified as bovine rhinovirus serotype 3. PMID- 2982347 TI - Isolation of ovine rotavirus in cell cultures. Brief report. AB - Three cytopathic rotavirus strains were isolated in MA 104 cells from intestinal contents of lambs with diarrhea. Pretreatment of virus with pancreatin and incorporation of a small amount of pancreatin in maintenance medium were important for establishment of the strains in these cells. The isolates showed marked cross reactions with bovine rotavirus in immunofluorescence, but no cross reaction with bovine, human, simian, equine, porcine and lapine rotaviruses in neutralization. PMID- 2982348 TI - Analysis of parapoxvirus genomes. AB - Eight stomatitis papulosa (SP), four orf and two milker's nodes (MN) virus isolates were compared by restriction enzyme analysis. Considerable genetic heterogeneity was found not only between isolates belonging to the three different taxonomic groups but also between members of the same group. This heterogeneity precludes classification of parapoxviruses simply by comparison of their DNA cleavage patterns. Restriction maps were therefore prepared for 12 parapoxvirus DNAs. Fragments from defined regions of the genome were then selected and used as probes for cross-hybridizations to all other parapoxvirus DNAs. DNA fragments derived from an internal region of the genome hybridized strongly to all parapoxvirus isolates examined. In contrast, cross-hybridization of the end region of the DNA molecule was observed only between members of the same virus group. Molecular hybridization as a means of classifying parapoxvirus isolates is discussed. PMID- 2982349 TI - Nucleic acid probes in diagnosis of viral diseases of man. Brief review. AB - With the recent, rapid advances in recombinant DNA technology, it has become possible to consider the use of nucleic acid probes in diagnosis of human viral diseases. Several examples are discussed which employ techniques of dot blot hybridization, sandwich hybridization and in situ hybridization. Typing of viral strains using restriction endonuclease digestion as an epidemiological tool is considered. Finally, the present limitations of molecular hybridization are discussed, and future developments including the production of non-radioactively labeled probes, are assessed. PMID- 2982350 TI - Experimental parainfluenza virus infection in mice: growth and spread of a highly pathogenic variant of parainfluenza 3 virus in the mouse brain. AB - We had previously showed that following intracerebral inoculation of newborn mice, the 910 N and M strains of bovine parainfluenza 3 virus induce a non-lethal hydrocephalus and a lethal disease with marked thymic and splenic atrophy, respectively. Moreover, only the M virus was lethal for 2-week-old mice. In the present study, we demonstrate that the M virus multiplies and spreads in the mouse brain invading the thalamus, hypothalamus and brain stem beyond the ependyma whereas the 910 N virus causes only slight ependymitis. This growth and spread of M virus was blocked by passive immunization 3 days after infection. Mouse embryo brain cell cultures were infected with M and 910 N viruses, about 50 per cent became antigen-positive for M whereas only a small proportion of cells were positive for the 910 N virus. However, the latter did produce higher yeilds of infectious virus than M. PMID- 2982351 TI - Isolation and characterization of cytopathic strains of rotavirus from rabbits. Brief report. AB - Three cytopathic rotavirus isolates were recovered from young rabbits affected by an enteric syndrome. The three isolates, when compared by cross serum neutralization tests, were found to be of the same serotype. Cross neutralization occurred also between a representative of the rabbit isolates and one strain of bovine rotavirus. PMID- 2982352 TI - The isolation of Meaban virus, a new Flavivirus from the seabird tick Ornithodoros (Alectorobius) maritimus in France. AB - Seven strains of a new Flavivirus, for which the name of Meaban virus is proposed, were isolated from the seabird tick Ornithodoros (A.) maritimus collected during 1981 and 1982 in nests of herring gulls (Larus argentatus) on islands of South Brittany, France. The new virus was compared serologically with 65 other flaviviruses including Tyuleniy virus and was found to be most closely related to, but different from Saumarez Reef virus, an agent previously isolated in Australia and Tasmania. Some general properties of Meaban virus are described and its antigenic relationships with other tick-borne flaviviruses associated with seabirds are discussed. PMID- 2982354 TI - Natural infection of Swiss mice with mouse mammary tumor virus (MMTV): viral expression in milk and transmission of infection. AB - Quantitative determinations of gp52, the main envelope glycoprotein, and p28, the main core protein, of MMTV, have been performed in about 1000 individual samples of milk of breeding females from our colony of MMTV-infected Swiss mice, a line characterized by a moderate incidence of mammary tumors. A computer analysis of the results showed: 1-- an important individual variation, ranging from 0 to 120 micrograms per ml of milk for p28, and from 0 to 320 micrograms per ml of milk for gp52; 2-- a variation of the release of both antigens during a single lactation, with a maximum on the 7--8th day of nursing; 3-- an increase of the release of both antigens with parity up to the 6th lactation, followed by a marked decrease during later lactations; 4-- a higher degree of infection in the offspring of 2nd and 3rd litters. The possible dependence of viral expression and transmission of infection upon factors such as cyclic activity of the mammary gland and progressive immunization of mice against MMTV is analyzed. The status of our laboratory line of MMTV infected Swiss mice is discussed in comparison with high and low tumor incidence strains. PMID- 2982353 TI - Rat cytomegalovirus induces cellular purine and pyrimidine nucleoside kinases in rat embryo fibroblasts and TK- rat-2 cells. Correlations with the antiviral activity of Acyclovir. AB - Rat cytomegalovirus (RCMV) induces a cytosol thymidine kinase (TK) in G0-phase rat embryo fibroblasts (REF), but not in a TK deficient rat cell line (R-2), though virus titers in both cell types reached comparable levels. The results indicate that TK is neither virus-coded nor is required for a productive infection in R-2 cells. A deoxycytidine kinase (dCK) is induced in either growing or RCMV-infected REF and R-2 cells, suggesting that dCK is essential for both host-cell and viral DNA synthesis. A deoxyguanosine kinase (dGK) is detectable in low concentrations in either growing or G0-phase REF and R-2 cells suggesting that this enzyme is cell-cycle independent. In contrast, RCMV induces high persisting levels of dGK, particularly in R-2 cells, indicating that this enzyme is of crucial importance for viral DNA synthesis. By comparison of thermostabilities and electrophoretic mobilities (Rf for TK, dCK and dGK were 0.12; 0.97; and 0.54, respectively) the enzymes were found to be substrate specific but of cellular origin. In contrast to TK and dCK, only dGK is inhibited by Acyclovir (Ki = 320 microM). It is suggested that RCMV inducable dGK is an important enzyme determining the in vitro anti-CMV activity of Acyclovir. PMID- 2982356 TI - Effect of staggered cyclophosphamide-immunosuppression on resistance to experimental Junin virus infection. AB - Otherwise resistant adult mice were rendered susceptible to intracerebral Junin virus (JV) infection only when a staggered cyclophosphamide (CY) schedule was used. Forty-five-day old Balb/c mice, intracerebrally JV-infected and immunosuppressed with four 50 mg/kg body weight CY doses at days -1, +1, +4, +6 (day 0: viral infection) developed a lethal disease (86.6 per cent mortality) with high CNS viral titers and brain lesions. Neutralizing antibodies were absent throughout, while immunofluorescent antibody levels were considerably diminished. The transfer of hyperimmune serum conferred partial though significant protection on CY-treated animals but no correlation was found between CNS viral titers and mortality since in both infected CY-treated and untreated mice similar brain viral content was found. This was also confirmed by immune spleen cell transfer at day 0 where the clearance achieved was unable to modify the time course of the disease. Feasible mechanisms explaining recovery from JV infection by means of the protective effect of antibodies and the cell-mediated clearance are discussed. PMID- 2982357 TI - Induction of Spodoptera littoralis nuclear polyhedrosis virus in cell cultures. AB - A nuclear polyhedrosis virus (NPV) infection was initiated in a Spodoptera littoralis cell line when haemocytes from granulosis virus (GV)-infected S. littoralis larvae and ultra-violet-inactivated Sendai virus were added. PMID- 2982355 TI - NMR parameters of local anesthetics as biological markers of the cell-virus interactions. AB - The selective proton NMR relaxation rates were measured in the system made by the local anesthetic procaine, human erythrocytes and the hemagglutinating Echo virus type 11. The NMR technique provided a very good tool for using drugs firmly bound to receptor membranes as biological "markers" of the virus-cell interaction. PMID- 2982358 TI - Studies on the development of the charge heterogeneity of the influenza virus glycoproteins. AB - The heterogeneity in charge of the influenza virus glycoproteins, hemagglutinin (HA) and neuraminidase (NA) is retained, when glycosylation is inhibited by tunicamycin (TM) or 2-deoxyglucose (2-dg). This is in contrast to the charge heterogeneity of the G protein of vesicular stomatitis virus (VSV), which is mainly due to heterogeneous sulfation of the carbohydrate side chains and therefore is abolished by the above mentioned inhibitors of glycosylation. Thus, the charge heterogeneity of influenza virus glycoproteins might be attributable to some as yet unidentified modifications of the polypeptide backbone. PMID- 2982359 TI - Herpes simplex virus latency in a hyperresistant clone of mouse neuroblastoma (Cl300) cells. AB - Herpes simplex virus (HSV) hyperresistant neuroblastoma cells (clone Cl300 RII) were latently infected with HSV-1 if cultured in presence of HSV-neutralizing antibody for one or two passages after infection and then passaged further without antibody. By superinfecting HSV-1 latently infected Cl300 RII cells with HSV-2, progeny virus with HSV-1 characteristics was regularly rescued. Such retrieval of HSV-1 decreased with passage of the latently infected cells. PMID- 2982360 TI - Role of Langerhans cells in the infection of the guinea-pig epidermis with foot and-mouth disease virus. AB - In guinea-pig infected with foot-and-mouth disease virus (FMDV), Langerhans cells in the foot pads increase in number and show viral antigens 24 hours post inoculation, preceding appearance of virus in epithelial cells and vesiculation. This observation suggests that Langerhans cells may be engaged in virus transport from the blood to the non vascularized epidermis. PMID- 2982361 TI - Influence of household handling. PMID- 2982362 TI - Purification of the cardiac 1,4-dihydropyridine receptor/calcium channel complex. AB - Chick heart membranes were labelled with [3H]PN 200-110, a 1,4-dihydropyridine calcium channel antagonist. The [3H]PN 200-110 receptors were solubilized with digitonin and purified 600-fold using several chromatographic systems and sucrose density gradient centrifugation. Analyses by gel electrophoresis revealed that the final product was enriched in two peptides with molecular weights of 60,000 and 54,000, and another of 34,000. PMID- 2982363 TI - Thiophosphorylation and phosphorylation of chromatin proteins from calf thymus in vitro. AB - Thiophosphorylation and phosphorylation of 5% perchloric acid extractable proteins from calf thymus chromatin were studied using a cyclic GMP-dependent protein kinase from bovine lung and a nuclear protein kinase II from rat liver. The phosphorylation reaction catalyzed by nuclear protein kinase II utilized [gamma -35S]ATP as a phosphate donor almost as efficiently as [gamma -32P]ATP, but the cGMP-dependent protein kinase mediated phosphorylation by [35S]ATP was about 20 times less effective than that by [32P]ATP. In addition, using [35S]ATP instead of [32P]ATP changed markedly the cGMP-dependent phosphorylation pattern of the PCA-extractable proteins as examined by gel electrophoresis. Thus, depending on the type of protein kinase, the results from thiophosphorylation and phosphorylation reactions may vary considerably. PMID- 2982364 TI - beta-Adrenergic activation of glycosyltransferases in the dolichylmonophosphate linked pathway of protein N-glycosylation. AB - beta-Adrenoreceptor stimulation of rat parotid acinar cells increases the activity of several microsomal membrane associated, dolichylmonophosphate (Dol-P) linked glycosyltransferases. The activities of Man-P-Dol synthase and Glc-P-Dol synthase are increased by approximately 50%, and the activity of N acetylglucosaminyl 1-phosphate transferase plus N-acetylglucosaminyl transferase increased by approximately 60%, after agonist treatment. Increases in enzyme activity are (i) independent of endogenous Dol-P levels and (ii) observed under conditions in which the specific activities of donor sugar nucleotides are kept constant. Activation of these enzymes is specific since comparable levels of NADPH-cytochrome c reductase are found in control and agonist-treated membranes. The data thus provide the initial demonstration of neurotransmitter modulation of enzymes in the dolichol-linked pathway of protein N-glycosylation. PMID- 2982365 TI - Identification of enzymatic activities which process protein bound mono(ADP ribose). AB - Enzymatic activities have been identified in extracts of cultured mouse cells which catalyze the removal of intact mono(ADP-ribosyl) residues linked to proteins at arginine. Activities that sequentially remove AMP and ribose 5 phosphate have also been identified. These results suggest that mono(ADP ribosylation) of proteins is a reversible post translational modification. PMID- 2982366 TI - Detection of a novel cyclooxygenase metabolite produced by human promyelocytic leukemia (HL-60) cells. AB - Arachidonic acid metabolism via the lipoxygenase pathway was examined in HL-60 cells before and after N,N-dimethylformamide induced differentiation along granulocytic lines. Untreated HL-60 cells produced small amounts of the 5 lipoxygenase products, 5-hydroxy-eicosatetraenoic acid and leukotriene B4 upon stimulation with calcium ionophore A23187. N,N-dimethylformamide treatment, caused a 10 to 20 fold increase in the amount of ionophore A23187-induced 5 lipoxygenase metabolites. An additional, and as yet unidentified arachidonic acid metabolite was routinely observed during reverse-phase high pressure liquid chromatography analyses of lipoxygenase products. Sensitivity to inhibition by less than 10(-7)M indomethacin coupled with other characteristics of its production, strongly suggest the compound is a cyclooxygenase product. The unusual UV absorbance and chromatographic elution pattern, however, suggest that it is not a typical prostaglandin, thromboxane or prostacyclin product. PMID- 2982368 TI - Inhibition of phosphatidylinositol-4-phosphate kinase by its product phosphatidylinositol-4,5-bisphosphate. AB - Phosphatidylinositol 4,5-bisphosphate (PIP2) is enzymatically produced when high speed supernatant fraction from bovine retina is incubated with [gamma-32P]ATP and phosphatidylinositol 4-phosphate (PIP) as substrates. Exogenously added PIP2 inhibits PIP kinase activity 50% at equimolar concentrations of product and substrate. Ca2+-dependent phosphodiesteratic activity, resulting in the loss of PIP2 and PIP and concommitant increase in myo-inositol 1,4,5-trisphosphate and myo-inositol 1,4-bisphosphate, was observed when soluble retinal fractions were incubated with heat-inactivated 32P-prelabeled guinea pig nerve ending membranes as substrate. It is suggested that polyphosphoinositides are under stringent and complex control and that upon receptor activation-mediated stimulation of phosphodiesteratic degradation release of the feedback inhibition shown here may occur and result in the synthesis and replenishment of PIP2. PMID- 2982367 TI - The expression of ceruloplasmin, an angiogenic glycoprotein, by mouse embryonic fibroblasts. AB - Balb/c 3T3, Swiss 3T3 and Rous sarcoma virus transformed Balb/c 3T3 mouse embryonic fibroblasts produced ceruloplasmin in vitro, whereas primary cultures prepared from the Balb/c mouse embryos did not produce ceruloplasmin. The amount of ceruloplasmin synthesis by the Balb/c 3T3 cell line is enhanced by Rous sarcoma virus-transformation (1.5-3 fold) and by treatment with dexamethasone (about 2.4 fold). The protein was identified as ceruloplasmin by immunoprecipitation with ceruloplasmin-specific polyclonal antibody, and by similarity of peptide maps, and subunit molecular weight (135,000 dalton) to that of authentic ceruloplasmin from primary cultures of mouse hepatocytes. PMID- 2982369 TI - A novel deletion in delta beta-thalassemia found in Japan. AB - High molecular weight DNA from a Japanese individual homozygous for delta beta thalassemia was analyzed by the blot hybridization technique of Southern. Results indicated a large deletion of the non-alpha-globin gene cluster, starting in the vicinity of 3' to the A gamma-globin gene and extending through the 3' side of the beta-globin gene. Persistent expression of the gamma-globin gene in adult life has been supposed to be caused by loss of a region located about 3-4 kb 5' to the delta-globin gene from comparison of the extents of deletions in several different forms of delta beta-thalassemia and HPFH (hereditary persistence of fetal hemoglobin). But the novel deletion found in the present case of delta beta thalassemia suggests that the above putative regulatory region does not have this effect on expression of the gamma-globin gene. Some explanations of expression of fetal type globin genes in this delta beta-thalassemia are discussed. PMID- 2982370 TI - A source of apparent pyrophosphate:fructose 6-phosphate phosphotransferase activity in rabbit muscle phosphofructokinase. AB - In the presence of UDPglucose, rabbit muscle phosphofructokinase appeared to use PPi as a phosphoryl donor, as reported previously (Biochem. Biophys. Res. Commun. 121, 842-847). This apparent activity was due to conversion of UDPglucose and PPi to glucose 1-phosphate and UTP, the latter being metabolized by phosphofructokinase. Auxiliary enzymes used in the assays were contaminated by UDPglucose pyrophosphorylase. This contamination was sufficient to account for, and had similar properties to, the apparent PPi-dependent activity. Without auxiliary enzymes phosphofructokinase could not use PPi. These findings indicate that the apparent interconversion of phosphofructokinase and PPi:fructose 6 phosphate phosphotransferase must be re-assessed. PMID- 2982371 TI - Specific binding of synthetic human pancreatic growth hormone releasing factor (1 40-OH) to bovine anterior pituitaries. AB - We have studied the specific binding of a synthetic 40 amino acid, free carboxy terminus analog of human pancreatic growth hormone releasing factor (hp GRF-40 OH) to partially purified homogenates of bovine anterior pituitaries. The binding of hpGRF-40-OH to pituitary receptors at 4 degrees C reached maximal level in 4 hours and remained steady for the next 18 hours. Specific binding increased linearly with the amount of protein present in the assay. 125I-hpGRF-40-OH binding to pituitary homogenates was competitively inhibited by hpGRF-40-OH but not by unrelated hormones. The competition curve and Scatchard analysis suggest the presence of single class of receptors with a Kd congruent to 3nM and binding capacity of approximately 200 fmoles/mg protein. This is the first demonstration of specific receptors for GRF on anterior pituitary cells. PMID- 2982372 TI - Prostaglandin E2 stimulates collagen and non-collagen protein synthesis and prolyl hydroxylase activity in osteoblastic clone MC3T3-E1 cells. AB - We investigated the stimulative effect of prostaglandin E2 (PGE2) on an osteoblastic cell line, clone MC3T3-E1, in serum-free medium. PGE2 elevated collagen and non-collagen protein syntheses in a dose-related fashion up to 2 micrograms/ml, the maximal increases being 2- and 3-fold, respectively, over that in the control. Its stimulative effect was evident as early as 12 h. PGE2 slightly increased DNA content, but its effect was less than that on collagen and non-collagen protein syntheses. Moreover, PGE stimulated an increase in prolyl hydroxylase activity with a maximal effect at 1-2 micrograms/ml, the activity being 15-fold over that of the control. These results strongly indicate that PGE2 directly enhances total protein synthesis including that of collagen in osteoblasts in vitro, suggesting its direct effect on bone formation in vivo as well. PMID- 2982374 TI - Requirements and functions of vesicular stomatitis virus L and NS proteins in the transcription process in vitro. AB - The L and NS proteins of vesicular stomatitis virus were purified from transcribing ribonucleoprotein complex and were used to study their requirements and functions during reconstitution of RNA synthesis in vitro. The requirements for L and NS proteins for optimal RNA synthesis were found to be catalytic and stoichiometric, respectively. Addition of increasing amounts of NS protein to N RNA template and saturating L protein, the ratio of N-mRNA to leader RNA synthesis increased linearly. In contrast, when the concentration of L protein was increased the corresponding ratio remained constant. These results, coupled with the observation that the L protein is involved in the initiation of RNA synthesis, suggest that the NS protein is involved in the RNA chain elongation step. The NS protein possibly interacts with both the L protein and the template N-RNA and unwinds the latter to facilitate the movement of L protein on the template RNA. PMID- 2982373 TI - Selective antiherpetic activity of carbocyclic analogues of (E)-5-(2 halogenovinyl)-2'-deoxyuridines: dependence on specific phosphorylation by viral thymidine kinase. AB - The carbocyclic analogues of (E)-5-(2-bromovinyl)-2'-deoxyuridine (C-BVDU) and (E)-5-(2-iodovinyl)-2'-deoxyuridine (C-IVDU), in which the sugar moiety is replaced by a cyclopentane ring, are as efficient substrates for the herpes simplex type 1 (HSV-1)-encoded thymidine kinase (TK) as their parent compounds (BVDU and IVDU). This conclusion is based on the binding affinities (Ki) of BVDU, IVDU, C-BVDU and C-IVDU to the HSV-1 TK and on the phosphorylation rates (Km, Vmax) of (125I)IVDU and (125I)C-IVDU by the enzyme. The specific phosphorylation of C-BVDU and C-IVDU by the viral TK may explain why these compounds are highly selective inhibitors of HSV-1 replication. PMID- 2982375 TI - Interaction of Leu-enkephalin with isolated enterocytes from guinea pig: binding to specific receptors and stimulation of cAMP accumulation. AB - The specific binding of Leu-enkephalin and the stimulatory effect of the peptide on cAMP accumulation have been assessed in isolated enterocytes of guinea pig. The binding was reversible as well as time and temperature dependent. Two classes of binding sites could be defined: a class with a relatively high affinity (Kd = 0.7 microM) that represented 1% of total binding capacity, and another class with low affinity (Kd = 55.5 microM). The stimulation of cAMP accumulation was also shown to depend on time and temperature and was potentiated by a phosphodiesterase inhibitor. Half-maximal stimulation of cAMP accumulation was observed at 119 microM and maximal stimulation (27-fold basal level) at 300 microM Leu-enkephalin. Both steps of the interaction were not modified by Na+ but exhibited a high specificity since modification in the structure of Leu enkephalin resulted in an important loss of binding affinity and stimulatory activity. PMID- 2982376 TI - Design of peptide derived amino alcohols as transition-state analog inhibitors of angiotensin converting enzyme. AB - A new amino alcohol modification designed to mimic the putative transition-state of amide bond cleavage by proteolytic enzymes has been incorporated into the scissile bond position of N-benzoyl-Phe-Ala-Pro, a known substrate of angiotensin converting enzyme (ACE). The resulting modified tripeptides (i.e. 4) are shown to be a new class of potent inhibitors of converting enzyme. PMID- 2982377 TI - Co-oxidation of luminol by hypochlorite and hydrogen peroxide implications for neutrophil chemiluminescence. AB - Stimulated neutrophils produce several potent oxidants including H2O2, O2- and HOCl. Previous studies have revealed all of these compounds to be capable of oxidizing luminol, a reagent often used to indicate, by its chemiluminescence, the oxidative burst of neutrophils. Data presented in this paper indicate that H2O2 and HOCl spontaneously react at physiologic pH to produce luminol-dependent chemiluminescence 100 times the sum of the chemiluminescence of either reagent alone. This enhancement is due to a co-oxidation by HOCl and H2O2, or to a novel oxidant generated by the interaction of HOCl and H2O2. The HOCl scavenger, taurine, inhibits the chemiluminescence. Evidence is presented against the participation of hydroxyl radical, O2- or singlet oxygen in the oxidation of luminol by HOCl and H2O2. These findings have implications for potential anti inflammatory compounds. PMID- 2982379 TI - Influence of fructose 2,6-bisphosphate on the phosphofructokinase/fructose 1,6 bisphosphatase cycle. AB - In a reconstituted enzyme system multiple stationary states and oscillatory motions of the substrate cycle catalyzed by phosphofructokinase and fructose 1,6 bisphosphatase are significantly influenced by fructose 2,6-bisphosphate. Depending on the initial conditions, fructose 2,6-bisphosphate was found either to generate or to extinguish oscillatory motions between glycolytic and gluconeogenic states. In general, stable glycolytic modes are favored because of the efficient activation of phosphofructokinase by this effector. The complex effect of fructose 2,6-bisphosphate on the rate of substrate cycling correlates with its synergistic cooperation with AMP in the activation of phosphofructokinase and inhibition of fructose 1,6-bisphosphatase. PMID- 2982378 TI - 24R,25-dihydroxyvitamin D3 regulates 1,25-dihydroxyvitamin D3 binding to its chick intestinal receptor. AB - We have studied the binding of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] to its crude chromatin chick intestinal receptor in the absence or presence of a ten fold excess of 24R,25-dihydroxyvitamin D3 [24R,25(OH)2D3] for each concentration of [3H]-1,25(OH)2D3 studied. We have found a significant shift to the right in the binding of 1,25(OH)2D3 to its receptor in the presence of this excess of 24R,25(OH)2D3. As a result, the affinity was found to be significantly reduced, the apparent dissociation constants varied from 0.97 +/- 0.09 (n = 5) to 1.36 +/- 0.04 nM (p less than 0.01). This reduction was related to a significant decrease in the positive cooperativity for the apparent Hill coefficient from nH = 1.49 +/ 0.06 to nH = 1.26 +/- 0.06 (p less than 0.03) in the binding of 1,25(OH)2D3 to its receptor. There was no significant change in the capacity of the receptor (189 +/- 11 compared to 200 +/- 9 fmoles/mg protein). These results suggest that the intestinal 1,25(OH)2D3 receptor must also have a binding recognition site for 24R,25(OH)2D3 which is postulated to play a regulatory role in the 1,25(OH)2D3 receptor's ligand binding properties. PMID- 2982380 TI - Purification and identification of endogenous anti-opioid substances from bovine brain. AB - Two fractions with high potency for reversing the inhibitory effect of Met enkephalin on the electrically induced contractility of guinea-pig ileum have been purified from bovine brain extract. Unexpectedly, one isolated peptide was identified as [Val5]-angiotensin I and the other fraction was [Val5]-angiotensin II, as judged by chromatographic comparisons on HPLC and amino acid analysis. Since angiotensins did not affect opioid binding to brain membrane, we consider that angiotensins may act as physiological antagonists to the opioid system in the brain, as well as in the guinea-pig ileum. PMID- 2982381 TI - Spin state studies on cytochrome P-450 in liver microsomes from obese and diabetic animals. AB - The spin state of liver microsomal cytochrome P-450 from obese mice and streptozotocin-diabetic mice and rats has been studied both by the temperature and the type I substrates-induced spectral changes. The high spin cytochrome P 450 is significantly decreased in these animals. Moreover absolute spectra indicate that low spin cytochrome P-450 is stabilized in streptozotocin induced diabetic animals. Thus the physiopathological state may modify the in vivo spin state of cytochrome P-450 and modifications of the microsomal fatty acid composition might contribute to these changes. PMID- 2982382 TI - Irreversible inactivation of rat gastric (H+-K+)-ATPase in vivo by omeprazole. AB - Subcutaneous administration of omeprazole, a gastric antisecretory agent belonging to the family of substituted benzimidazoles, brought about a dose dependent decrease in gastric mucosal (H+-K+)-ATPase activity in the rat. The dose which inhibited 50% of the enzyme activity was 1 mg/kg from dose-response profiles obtained 3 h after the drug dosing. Duration profiles of the drug at 10 mg/kg showed that its ATPase-inhibitory effect reached the maximum in 2 h with 80% reduction of the enzyme activity. The gastric mucosal level of the ATPase activity remained to be maximally inhibited for 12 h and returned to a normal level with a half time of about 20 h. The return of the enzyme activity, however, was blocked by treatment with cycloheximide, an inhibitor of protein synthesis. These observations indicate that omeprazole irreversibly inactivates gastric (H+ K+)-ATPase in vivo. PMID- 2982383 TI - Epidermal growth factor inhibits growth while increasing the expression of an ecto-Ca2+-ATPase of a human hepatoma cell line. AB - We have obtained a cloned cell line (Li-7A) from primary cultures of a human hepatoma xenograft (Li-7). Li-7A was able to grow in the absence of serum. Growth was stimulated 0-3 fold by addition of newborn calf serum, but was inhibited in DME/F12 media containing nine growth factors. The ectoMg2+-ATPase was 1.5-2 fold higher than the ectoCa2+-ATPase activity in cells grown in media with or without serum. In cells grown in media supplemented with the nine factors, the ectoCa2+ ATPase activity exceeded the ectoMg2+-ATPase, and there was also a 5-10 fold increase in its specific activity. Inhibition of growth was due to epidermal growth factor alone. The increased expression of the ectoCa2+-ATPase was absolutely dependent on EGF, but also required hydrocortisone and cholera toxin. The characteristics of Li-7A cells make it a suitable system for studying both the mechanism of action of EGF and plasma membrane ATPases. PMID- 2982384 TI - Prostaglandins and cyclic nucleotides. Modulators of arterial cholesterol metabolism. PMID- 2982385 TI - The oxidation of p-phenetidine by horseradish peroxidase and prostaglandin synthase and the fate of glutathione during such oxidations. AB - The oxidation of p-phenetidine by horseradish peroxidase and prostaglandin synthase was investigated. The existence of a free radical intermediate formed during enzymatic oxidation was supported by a ratio of hydrogen peroxide: p phenetidine consumed of 1:2 in the horseradish peroxidase system. Furthermore in both enzyme systems a rapid oxidation of added glutathione was observed and in the presence of the thiol there was a decreased removal of p-phenetidine. This suggests the reduction of a p-phenetidine radical by glutathione generating p phenetidine and a thiyl radical. The latter react with oxygen and a rapid oxygen uptake was observed during enzymic oxidation in the presence of thiols. That p phenetidine radicals were produced during horseradish peroxidase catalyzed oxidation of p-phenetidine was supported by experiments using the spin probe OXANOH. This was oxidized to its stable free radical form (OXANO.) in an enzyme- and substrate-dependent reaction and the EPR signal obtained was not decreased by SOD (80 micrograms/ml) or benzoate (10-100 mM). TLC characteristics of the products of the oxidation of p-phenetidine by both enzymes were almost identical inferring a similar mechanism of oxidation. Two of the metabolites were characterized by mass spectrometry and by comparison with reference compounds prepared by chemical oxidation. One metabolite was identified as 4,4' diethoxyazobenzene, which further supports a radical mechanism, and the other was a p-phenetidine trimer which could exist in both oxidized and reduced forms. On the basis of these observations a mechanism for the oxidation of p-phenetidine and the fate of glutathione during such oxidations is proposed. PMID- 2982386 TI - Superoxide radical reactions with anthracycline antibiotics. AB - The reaction of superoxide with daunorubicin or its aglycones in the aprotic solvents dimethyl sulfoxide and dimethylformamide was studied. This interaction generated the blue anthracycline phenolate anion as monitored by u.v.-visible spectrometry and molecular oxygen as determined by a modified Clark-type oxygen electrode. The visible spectrum of the phenolate anion (gamma max 604, 652 nm) was subject to considerable shifts dependent on the size of the cation present. The phenolate anion could be further oxidized by molecular oxygen to generate the C-6, C-11 (B-ring) semiquinone as detected by a weak electron paramagnetic resonance spectrometry signal. These results raise the possibility that similar reactions of superoxide with anthracyclines in vivo may play a role in the antitumor activity and/or the etiology of the toxic side effects of this class of drugs. PMID- 2982387 TI - Effect of induction of anaesthesia with etomidate on corticosteroid synthesis in man. AB - The effects of a single bolus dose of etomidate 0.3 mg kg-1 or thiopentone 5 mg kg-1, on the synthesis of corticosteroid hormones and adrenocorticotrophic hormone (ACTH), were compared for 24 h in 12 patients, undergoing minor surgery under general anaesthesia. Following opioid premedication i.m. and general anaesthesia, plasma cortisol concentrations decreased transiently within the first hour of anaesthesia in all 12 patients. The six patients who received etomidate had statistically higher plasma 11-deoxycorticosterone concentrations at 4 and 24 h than those who had received thiopentone (P less than 0.01). Throughout the study, no difference in plasma cortisol, corticosterone or ACTH concentrations were found between the two groups. We have demonstrated a biochemical effect of a single bolus dose of etomidate consistent with incomplete inhibition of adrenocortical mitochondrial 11 beta-hydroxylase activity, but no clinically significant adrenocortical suppression. PMID- 2982388 TI - Clinical actions of fentanyl and buprenorphine. The significance of receptor binding. AB - Receptor binding assays were undertaken in an attempt to elucidate the opioid binding characteristics of fentanyl and buprenorphine, and to investigate some of the differences between them. Buprenorphine showed slow receptor association (30 min), but with high affinity to multiple sites from which dissociation was very slow (T 1/2 = 166 min) and incomplete (50% binding after 1 h). This contrasted with the receptor binding of fentanyl, which achieved rapid equilibrium (within 10 min) and dissociated equally rapidly (T 1/2 = 6.8 min) and completely (100% by 1 h). Competitive displacement showed buprenorphine displacement of fentanyl binding was concentration- and time-dependent over ranges encountered in clinical use, but buprenorphine binding was displaced with only very high concentrations of other opioids. These findings offer pharmacodynamic explanations for the differences in fentanyl and buprenorphine analgesic response profiles and suggest how binding interactions might be applied to therapeutic use. PMID- 2982389 TI - Increased degranulation of human myeloperoxidase-deficient polymorphonuclear leucocytes. AB - Myeloperoxidase (MPO)-deficient neutrophils (PMN) released considerably more beta glucuronidase, lysozyme and vitamin B12-binding activities, when exposed to opsonized zymosan (STZ), than the normal counterpart. Release of the soluble enzyme lactate dehydrogenase was not appreciably changed over the incubation time with particles in either cell type. MPO-deficient PMN and normal PMN ingested STZ particles at a similar rate at early times, but thereafter phagocytosis by MPO deficient PMN was significantly higher than that by normal PMN. The difference in degranulation between the two cell types greatly exceeded the difference in ingestion and was evident already at early phagocytosis times when no difference in phagocytosis was observed; this suggested that the higher degranulation in MPO deficient PMN was at least in part independent of the increased ingestion. This was confirmed by experiments with the soluble stimulant N-formyl-L-norleucyl-L leucyl-phenylalanine (FNLLP). MPO-deficient PMN and normal PMN exhibited a comparable respiratory burst when exposed to FNLLP plus cytochalasin B, but the defective cells released more azurophilic and specific granule markers than normal PMN. These results indicate that MPO-deficient PMN degranulate more than normal PMN and suggest a role for MPO in the regulation of degranulation. PMID- 2982390 TI - Pathogenesis of hepatic fibrosis in experimental iron overload. AB - Significant increases in prolyl hydroxylase activity, a key enzyme in the collagen biosynthetic pathway, were noted in the hepatic homogenates of iron loaded animals as compared to controls. The increase in prolyl hydroxylase activity was seen without any light microscopic histologic evidence of cell necrosis or accumulation of collagen in the livers from the iron-loaded animals. However, utilizing electron microscopy, collagen fibrils were demonstrated immediately adjacent to the hepatocytes in the iron-loaded animals but not the controls. No fibroblasts or inflammatory cells were noted in this area. There was no evidence of damage to the subcellular organelles of the iron-loaded hepatocytes. The hydroxyproline content of the iron-loaded livers was also shown to be increased. These experimental studies in conjunction with clinical observations described below suggest that in vivo iron overload may have a primary effect on stimulating collagen synthesis by hepatocytes. PMID- 2982391 TI - The storage pool deficiency in platelets from humans with the Chediak-Higashi syndrome: study of six patients. AB - Functional and biochemical studies of platelets from human Chediak-Higashi syndrome (CHS) are scarce and/or incomplete. In the present report, the aggregation response to a variety of inducers of platelet aggregation, the content of the dense granule constituents ATP, ADP, serotonin and calcium, the secretion of ATP, ADP, and calcium induced by thrombin, the total content of magnesium, the incorporation of 14C-adenine in the cytoplasmic pool of adenine nucleotides, as well as the content of intracellular cyclic-AMP, have been quantitated in six patients with CHS. Furthermore, data is presented on the kinetics of uptake of radiolabelled serotonin and its storage in human CHS platelets. An abnormal aggregation behaviour was found in all patients. However, the response of CHS platelets to the different inducers studied did not show a uniform pattern. The total content and the maximal amounts of the dense granule constituents secretable by thrombin were greatly decreased in all six patients. Total magnesium content was similar to that of normal platelets. The ATP/ADP ratio was higher than in controls. Uptake of radiolabelled serotonin by CHS platelets closely followed the uptake by normal platelets; during the first 2-3 min, however, incorporation of the amine by CHS platelets came rapidly to a plateau which contrasts with the steady, linear increase in uptake found in controls. CHS platelets loaded with radiolabelled serotonin and gel-filtered, showed a spontaneous release of radioactivity not observed in normal platelets under the same conditions. The cyclic-AMP content of CHS platelets was similar to that of normals. In contrast to platelets from patients with storage pool disease, the secretable calcium from CHS platelets represents a 67% of total platelet calcium (61% in normals), suggesting that the absolute values for the non-secretable portion in CHS platelets must be very low. The results reported confirm the existence of a true storage pool deficiency of the dense granule constituents as a common defect in CHS platelets. The variety of responses among patients, to the different aggregatory stimuli studied, can not be solely ascribed to the storage pool deficiency described. PMID- 2982393 TI - Respiration induces variable porosity to polyols in the mitochondrial inner membrane. AB - The osmotic basis of low and high amplitude swelling in mitochondria was investigated in detail using sucrose and mannitol as external osmolytes. Osmotic behaviour of mitochondria in various respiratory states was consistent with significant changes in the porosity of the inner membrane corresponding to the rate of respiration. The stoichiometry of oxidative phosphorylation was confirmed to be dependent on the physical state (i.e., osmotic stretch) of the inner membrane regardless of the external polyol used. High amplitude swelling in polyol media was shown to arise from a sequential disruption of the outer and inner mitochondrial membranes, due to a dynamic instability induced by a combination of respiration, unscreened (fixed) surface charge density and the consequent variable porosity of the inner membrane. These novel experimental findings based on the physical theory of osmosis emphasize the need to define the fine structural changes of the inner membrane associated with oxidative phosphorylation to arrive at a comprehensive mechanism. PMID- 2982394 TI - NMR and kinetic characterization of the interaction between French bean plastocyanin and horse cytochrome c. AB - French bean plastocyanin is shown by stopped-flow kinetics to oxidize horse cytochrome c with k (298 K, I = 0.10 M) = 5.1 X 10(6) M-1 X s-1. The activation parameters demonstrate a satisfactory isokinetic correlation with those previously reported for plastocyanin-cytochrome f reactions. NMR line broadening and shifts of the hyperfine shifted resonances of cytochrome c(III) reveal that strong 1:1 complexes are formed with plastocyanin. The negative patch of plastocyanin and the heme edge region of cytochrome c are shown to be the interacting sites by the hyperfine shift perturbations and competitive binding experiments with Gd3+, which associates selectively with the negative patch of plastocyanin. Complexation of plastocyanin and cytochrome c causes a small change in the heme electronic structure, but there is no NMR or optical evidence for significant conformational changes at either metal center. The rate of the reverse electron-transfer reaction within the plastocyanin-cytochrome c complex has been directly measured by NMR line broadening (krev (298 K) = 87 s-1). A rate for the forward intracomplex electron-transfer reaction (kf (298 K) = 4.8 X 10(3) s-1) has been calculated from krev and the optically measured equilibrium constant. PMID- 2982392 TI - Respiratory abnormalities among workers in an iron and steel foundry. AB - A study of the health of 78 workers in an iron and steel foundry in Vancouver, British Columbia, was carried out and the results compared with those found in 372 railway repair yard workers who were not significantly exposed to air contaminants at work. The foundry workers were exposed to PepSet, which consists of diphenyl methane diisocyanate (MDI) and phenol formaldehyde and their decomposition products as well as to silica containing particulates. A questionnaire was administered by trained interviewers, and chest radiography, allergy skin tests, pulmonary function tests, and methacholine inhalation tests were carried out as well as measurement levels of dust and MDI. Compared with the controls, the foundry workers had more respiratory symptoms and a significantly lower mean FEV1 and FEF25-75% after adjustments had been made for differences in age, height, and smoking habit. Three workers (4.8%) had radiographic evidence of pneumoconiosis and 12 (18.2%) had asthma defined as presence of bronchial hyperreactivity, cough, and additional respiratory symptoms such as wheeze, chest tightness, or breathlessness. Sensitisation to MDI is probably the cause of asthma in these workers. PMID- 2982395 TI - The acceptor quinone complex of Rhodopseudomonas viridis reaction centers. AB - The acceptor complex of isolated reaction centers from Rhodopseudomonas viridis contains both menaquinone and ubiquinone. In a series of flashes the ubiquinone was observed to undergo binary oscillations in the formation and disappearance of a semiquinone, indicative of secondary acceptor (QB) activity. The oscillating signal, Q-B, was typical of a ubisemiquinone anion with a peak at 450 nm (delta epsilon = 6 mM-1 X cm-1) and a shoulder at 430 nm. Weak electrochromic bandshifts in the infrared were also evident. The spectrum of the reduced primary acceptor (Q-A) exhibited a major peak at 412 nm (delta epsilon = 10 mM-1 X cm-1) consistent with the assignment of menaquinone as QA. The Q-A spectrum also had minor peaks at 385 and 455 nm in the blue region. The same spectrum was recorded after quantitative removal of the secondary acceptor, when only menaquinone was present in the reaction centers. Spectral features in the near-infrared due to Q A were attributed to electrochromic effects on bacteriochlorophyll (BChl) b and bacteriopheophytin (BPh) b pigments resulting in a distinctive split peak at 810 and 830 nm (delta epsilon = 8 mM-1 X cm-1). The menaquinone was identified as 2 methyl-3-nonylisoprenyl-1,4-naphthoquinone (menaquinone-9). The native QA activity was uniquely provided by this menaquinone and ubiquinone was not involved. QB activity, on the other hand, displayed at least a 40-fold preference for ubiquinone (Q-10) as compared to menaquinone. Thus, both quinone-binding sites display remarkable specificity for their respective quinones. In the absence of donors to P+, charge recombination of the P+Q-A and P+Q-B pairs had half-times of 1.1 +/- 0.2 and 110 +/- 20 ms, respectively, at pH 9.0, indicating an electron-transfer equilibrium constant (Kapp2) of at least 100 for Q-AQB in equilibrium QAQ-B. Also observed was a slow recombination of the cytochrome c 558+ Q-A pair, with t 1/2 = 2 +/- 0.5 s at pH 6. PMID- 2982397 TI - Structure-function studies of canine cardiac sarcolemmal membranes. I. Estimation of receptor site densities. AB - A novel method for the estimation of receptor site densities in purified canine cardiac sarcolemmal vesicles is described. Canine sarcolemmal vesicles, purified by the method of Jones et al. (Jones, L.R., Maddock, S.W. and Besch, H.R. (1980) J. Biol. Chem. 255, 9971-9980) had high (Na+ + K+)-ATPase specific activity (127 +/- 1.9 mumol Pi/mg per h). Total phospholipid content, estimated by measurements of total phosphorus and total fatty acid contents, was 3.09 mumol/mg. Saturation isotherms for several receptor ligands gave the following values for Kd and Bmax: ouabain 32.6 +/- 2.7 nM, 365 +/- 59 pmol/mg; quinuclidinyl benzilate 0.055 +/- 0.010 nM, 5.8 +/- 0.7 pmol/mg; dihydroalprenolol 4.6 +/- 1.0 nM, 2.2 +/- 0.2 pmol/mg; and nitrendipine 0.21 +/- 0.04 nM, 0.93 +/- 1.04 pmol/mg. Membrane phospholipid surface area per ligand-binding sites was estimated from the Bmax values for each receptor ligand utilizing 3.09 mumol phospholipid/mg and 60 A2 as the average surface area occupied by each phospholipid molecule. The following receptor site densities per micrometer 2 phospholipid surface were obtained: ouabain, 400; quinuclidinyl benzilate, 6; dihydroalprenolol, 2; and nitrendipine, 1. As the surface area contributed by protein was estimated to be less than 20% of the lipid surface area, these values must be reduced by approx. 20% to estimate site densities per micrometer 2 membrane surface. These data demonstrate much lower beta-adrenergic and muscarinic receptor density compared to that of Na+ pump sites. PMID- 2982398 TI - Studies on protein-lipid interactions in cytochrome c oxidase by differential scanning calorimetry. AB - The interaction between cytochrome c oxidase and phospholipids was studied by differential scanning calorimetry. The active, lipid-sufficient cytochrome c oxidase undergoes thermodenaturation at 336 K with a relatively broad and concentration dependent endothermic transition. The delipidated enzyme shows an endothermic denaturation temperature at 331.3 K. When the delipidated cytochrome c oxidase was treated with chymotrypsin, a lowered thermodenaturation temperature was observed. When the delipidated cytochrome c oxidase was reconstituted with asolectin to form a functionally active enzyme complex, the thermodenaturation shifted to a higher temperature, with a sharper transition thermogram. The increase in thermotransition temperature and enthalpy change of thermodenaturation of the asolectin-reconstituted enzyme is directly proportionate to the amount of asolectin used, up to 0.5 mg asolectin per mg protein. The thermotransition temperature and enthalpy changes of thermodenaturation for the phospholipid-reconstituted cytochrome c oxidase are affected by the phospholipid headgroup and the fatty acyl groups. Among phospholipids with the same acyl moiety but different head groups, phosphatidylethanolamine was found to be more effective than phosphatidylcholine in protecting cytochrome c oxidase from thermodenaturation. An exothermic transition thermogram was observed for delipidated cytochrome c oxidase embedded in phospholipid vesicles formed with phospholipids containing unsaturated fatty acyl groups. The increase in exothermic transition temperature and exothermic enthalpy change of thermodenaturation of the oxidase-cytochrome c-cytochrome c oxidase complex destabilized cytochrome c but not cytochrome c oxidase toward thermodenaturation. PMID- 2982396 TI - The interaction of yeast Complex III with some respiratory inhibitors. AB - We have examined the effects of eight inhibitors of the bovine-heart mitochondrial Complex III on the catalytic activity of the analogous complex from yeast mitochondria. All eight compounds were inhibitory, with potent inhibition being obtained with antimycin, myxothiazol and UHDBT (5-N-undecyl-6-hydroxy-4,7 dioxobenzothiazole). These three inhibitors, and also funiculosin, have been further studied by characterizing their effects on the visible absorbance, magnetic circular dichroism and EPR spectra of the complex and also on the potentiometric properties of the individual metal centers present in the complex. All four inhibitors had little or no effect on either the absorbance or magnetic circular dichroism spectra. Funiculosin produced a change in the EPR lineshape of the iron-sulfur cluster; EPR spectra recorded at 12 K also revealed complete reduction of cytochrome b-562 by ascorbate. UHDBT also changed the lineshape of the iron-sulfur cluster and this change could be partially reversed by myxothiazol. Neither antimycin nor myxothiazol affected the iron-sulfur cluster and produced only small changes in the EPR absorption envelope of the b cytochromes. Both funiculosin and UHDBT raised the midpoint potential of the iron sulfur cluster, by about 150 and 70 mV, respectively. Only UHDBT changed the potential of c1, lowering it by about 30 mV. Funiculosin raised the potential of b-562 by about 30 mV, while myxothiazol had no effect; the other two compounds produced only small changes. All four compounds had only small effects on the midpoint potential of b-566. The relative contributions of the two b cytochromes to the magnetic circular dichroism amplitudes could be changed by the addition of inhibitors, even though the absolute magnetic circular dichroism spectra of oxidized and reduced complex were unaffected. PMID- 2982399 TI - Binding of prostaglandin E1 to human erythrocyte membrane. AB - Prostaglandin E1 is known to alter the structural and functional characteristics of red blood cells, yet, little is understood about the membrane receptors mediating this process. We therefore studied the binding of tritium-labeled prostaglandin E1 to the intact human erythrocyte membrane and demonstrated that the interaction is highly specific, rapid, saturable and reversible. Scatchard analysis of prostaglandin E1 binding to the membrane preparations showed the presence of two independent classes of prostaglandin E1 binding sites which differed in their affinity for the autacoid. The high-affinity class had Kd = 3.6 X 10(-9) M and the low-affinity class had Kd = 5.6 X 10(-5) M. The optimum pH for the binding of [3H]prostaglandin E1 to the erythrocyte membrane was found to be around 7.5 and maximum specific binding occurred at a concentration of 5 mM Mg2+ in the incubation mixture. [3H]Prostaglandin E1 bound to the membrane preparation could not be displaced by GTP or by its stable derivative Gpp[NH]p. However, prostaglandin E1 bound to the erythrocyte membrane preparation could be rapidly displaced by cyclic AMP. The IC50 (concentration of the nucleotide displacing 50% bound [3H]prostaglandin E1 from the membrane) was 75 nM. Other adenine nucleotides or cyclic GMP could not substitute for cyclic AMP. Unlike the right side-out erythrocyte membrane, the inside-out membrane preparations do not bind [3H]prostaglandin E1. Treatment of right-side-out erythrocyte membrane preparation with neuraminidase markedly decreases the binding of prostaglandin E1. Incubation of the erythrocyte membrane preparation with trypsin resulted in total loss of the binding activity. These results indicate that the prostaglandin E1 binding sites located on the cell surface and sialic acid residues are required for prostaglandin E1 binding to the human erythrocytes. These results also indicated that the binding sites are glycoprotein in nature. PMID- 2982400 TI - Evidence against an abnormal hepatic microsomal lipid matrix as the primary genetic defect in the jaundiced Gunn rat. AB - The congenitally jaundiced Gunn rat does not conjugate bilirubin but does conjugate bilirubin dimethyl diester. Partial defects in conjugating p nitrophenol and demethylating aminopyrine are also evident. A proposed mechanism to explain this combination of findings is a defective microsomal membrane. To examine the 'matrix' of Gunn microsomal membranes, hepatic microsomes were isolated from Gunn (jj) and outbred Wistar (JJ) rats and were studied by electron paramagnetic resonance spectroscopy of 7-doxylstearic and 12-doxylstearic acid probes, fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene, glucose-6 phosphatase activity vs. temperature, and lipid analysis. The data indicate several factors related to lipid bilayer order do not differ in microsomes from jj and JJ. PMID- 2982401 TI - Strong interactions between a spin-labeled cholesterol analog and erythrocyte proteins in the human erythrocyte membrane. AB - We have used a spin label analog of cholesterol bearing a nitroxide on the alkyl chain (26-nor-25-doxylcholestanol) to study cholesterol-protein interactions in the human erythrocyte membrane. As judged from the ESR spectrum, the spin label is readily incorporated into the membrane when added from a concentrated ethanolic solution to a cell or ghost suspension. With intact erythrocytes or white ghosts in isotonic buffer, the ESR spectrum is a superposition of a mobile component and a strongly immobilized component (outer hyperfine splitting 61-63 G). The latter corresponds to approx. 45% of the signal, a percentage which is barely affected by varying the temperature between 5 and 37 degrees C. Removal of the cytoskeletal proteins spectrin and actin by low ionic strength treatment or of all extrinsic proteins by alkali treatment of ghosts reduces the immobilized fraction to approx. 25%. The effect of controlled proteolysis of intrinsic proteins was also tested. Pre-treatment of cells with chymotrypsin or pre treatment of unsealed ghosts with trypsin has no effect on the ESR spectrum obtained with alkali-treated membranes. On the other hand, after chymotrypsin treatment of unsealed ghost, which reduces the band 3 protein to a 17.5 kDa membrane fragment, the strongly immobilized component is no longer observable. These data show that the cholesterol analog 26-nor-25-doxylcholestanol interacts strongly with one or several proteins of the erythrocyte membrane. That the intrinsic protein band 3 is involved is suggested by the disappearance of the immobilized fraction occurring upon chymotrypsin digestion of this protein. Our results are thus consistent with the proposal of a selective cholesterol-band 3 interaction in the erythrocyte membrane (Schubert, D. and Boss, K. (1982) FEBS Lett. 150, 4-8). Our data also suggest that this interaction is influenced by cytoskeletal proteins, an effect which can be explained considering the known linking of band 3 to the erythrocyte cytoskeleton via ankyrin. Experiments have also been carried out with 3-doxylandrostanol, a more commonly used cholesterol spin-label analog. With this spin label, at all temperatures investigated, we found it impossible to demonstrate unambiguously the existence of two spectral components. It is suggested that 26-nor-25-doxylcholestanol is a better reporter of cholesterol behavior in membranes. PMID- 2982403 TI - Sensitivity and reversibility of Ca-dependent inhibition of the (Na+ + K+)-ATPase of human red blood cells. AB - The sensitivity of the (Na+ + K+)-ATPase to inhibition by Ca was increased 30 fold by a partially purified extract of human red cell hemolysate. The hemolysate fraction reduced the concentration of free Ca required for 50% inhibition from 30 microM to approx. 1 microM. Ca-dependent inhibition of the (Na+ + K+)-ATPase in the presence and absence of the hemolysate fraction was completely reversible. The hemolysate fraction also stimulated the Ca2+-ATPase and increased its affinity for Ca. In the presence of the hemolysate fraction, the concentration of free Ca that inhibited the (Na+ + K+)-ATPase by 50% was similar to that which half-maximally stimulated the Ca2+-ATPase. Boiling the fraction destoryed its effect on the (Na+ + K+)-ATPase, but did not impair its stimulation of the Ca2+ ATPase. PMID- 2982402 TI - Isolation of plasma membranes from the bovine retinal pigment epithelium. AB - Retinal pigment epithelium plasma membranes have been isolated by differential and density gradient centrifugation of glass-bead-bound, collagenase-treated cells. Electron microscopic evidence indicates that the glass-bead-bound cells were devoid of red blood cells, rod outer segments and other ocular cell contaminants. The plasma membranes were recovered in 4-6 micrograms/eye yields and purified 10-fold by 5'-nucleotidase and alkaline phosphodiesterase I, and 6.5 fold by (Na+ + K+)-ATPase. Plasma membrane purity as measured by covalent labeling of the epithelial cell plasma membrane proteins with p-(diazonium) benzene[32S]sulfonic acid was 8-19-fold. In purified plasma membranes contamination by mitochondria was undetectable and lysosomal contamination reduced 100-fold, while endoplasmic reticulum was 2-fold enriched. SDS polyacrylamide gel electrophoresis of the plasma membrane proteins revealed 23-26 major bands by Coomassie blue staining and 12-16 major bands by radioactive labeling. The plasma membranes exhibited a 3-fold lower concentration of docosahexaenoic acid, a 3-fold higher cholesterol/phosphate ratio, and were 10 fold enriched in cholesterol per micrograms protein when compared to the whole cell fraction. Retinal epithelial plasma membranes contain an average of 1 mol cholesterol per mol of lipid phosphorus, a high palmitic acid concentration (39 mol%) and a low concentration of docosahexaenoic acid (2 mol%). The lipid profile of the retinal pigment epithelial plasma membranes indicates that they are typical of plasma membranes from many other cell types and that they appear to be less fluid than total rod outer segment membranes. PMID- 2982404 TI - Characterization of the feline c-abl proto-oncogene. AB - Analysis of total feline DNA by genomic blot hybridization, using the viral oncogene of Abelson murine leukemia virus as a specific probe, has led to the identification of multiple v-abl homologous genetic sequences in the cat genome. Upon restriction endonuclease BamHI digestion, the combined size of the v-abl homologous DNA fragments was about 31 kbp. To characterize these sequences further, four independent v-abl homologous cosmid clones with overlapping cellular inserts have been isolated from a gene library of cat lung genomic DNA. These inserts represent a contiguous region of cellular DNA sequences of 56 kbp in length. Within this region of the feline genome, the v-abl homologous sequences are discontinuously dispersed over a region of about 34 kbp. They represent the complete feline v-abl cellular homolog and are colinear with the viral v-abl oncogene. Nine regions of highly repetitive DNA sequences have been mapped in close proximity to v-abl homologous sequences. These results establish the presence of only a single c-abl proto-oncogene in the cat genome and present its genetic organization. PMID- 2982405 TI - The effect of 2'-fluoro-2'-deoxycytidine on herpes virus growth. AB - The effect of 2'-fluoro-2'-deoxycytidine (dCfl) on the growth of certain viruses of the herpes type was investigated. It is shown that the compound has considerable anti-viral activity against HSV-I, HSV-II, pseudorabies virus and equine abortion virus. It has an effect comparable to that of araC and is more efficient than br5dC, but less so than acyclovir. Experiments with thymidine kinase-negative strains of HSV-I indicated that dCfl was phosphorylated by the viral kinase, and its Km appears to be low and close to that of thymidine. Density gradient centrifugation enabled us to show that dCfl was incorporated into cellular and viral DNA and RNA. The cytotoxic activity of dCfl appears to be about 10-times smaller than that of araC. Removal of the nucleoside analog, washing and replacement with deoxycytidine reversed this effect, indicating rather a cytostatic than cytotoxic effect. PMID- 2982407 TI - Proton Overhauser experiments on kringle 4 from human plasminogen. Implications for the structure of the kringles' hydrophobic core. AB - 1H-NMR Overhauser experiments at 300 and 600 MHz have been implemented on the isolated kringle 4 fragment of human plasminogen. This study shows that Leu46 and Leu77 CH3 delta,delta' groups, as well as two threonine CH3 gamma and a methionine S-CH epsilon (probably Met48) groups, are in efficient dipolar contact with histidine and aromatic side-chains. In particular, the experiments reveal that of the two Leu46 CH3 delta,delta' groups, one is in efficient contact with tryptophan (Trp25 and Trp62) indole rings while the other interacts with a tyrosine (probably Tyr41) phenol. Leu46 appears also to be close to an Ala CH3 beta group. Such a hydrophobic cluster appears to be contiguous to Trp72, hence to Arg71, residues that are through to be part of the lysine-binding site. Acid base titration experiments show that the buried methionine S-CH3 epsilon group senses a neighboring ionizable group of pK*1 = 3.76, suggesting presence of a carboxyl anionic group (probably an aspartic acid side-chain) in the vicinity of the hydrophobic core. A preliminary model is proposed for the overall folding of the kringle polypeptide chain. PMID- 2982406 TI - Quantitation of eukaryotic topoisomerase I reactivity with DNA. Preferential cleavage of supercoiled DNA. AB - A method has been used to quantitate the reaction between eukaryotic type I DNA topoisomerase and topological forms of DNA. This procedure (Trask, D.K., DiDonato, J.D. and Muller, M.T. (1984) Eur. Mol. Biol. Organ. J. 3, 671-676) measures the efficiency of DNA cleavage and concurrent formation of a covalent enzyme/DNA complex. Eukaryotic type I topoisomerases react preferentially by 5-10 fold with supercoiled DNA. The effect of supercoiling is clearly evident in that both the initial rate and final extent of the reaction is elevated. Because the dissociation rate is much lower than the association rate, it is possible to isolate native topoisomerase/DNA complexes. These complexes are comprised of enzyme molecules which are catalytically active when challenged with a second supercoiled DNA substrate. Collectively, the data support the conclusion that a functional intermediate in the reaction sequence is being detected and that the avian topoisomerase I preferentially cleaves supercoiled DNA. PMID- 2982408 TI - Interaction of oxidized chicken ovotransferrin with chicken embryo red blood cells. AB - Iron-saturated chicken ovotransferrin was chemically oxidized with NaIO4, converting 50% of its methionine residues to their sulfoxide derivatives while maintaining 95% of its iron-binding activity. The oxidized chicken ovotransferrin was able to deliver iron to the chicken embryo red blood cell for heme synthesis. From competition experiments, oxidized diferric chicken ovotransferrin was estimated to be approx. 65% as efficient as unmodified diferric chicken ovotransferrin at competing with diferric (55Fe2) chicken ovotransferrin for the iron-donating sites of the chicken embryo red blood cells. The presence of apo chicken ovotransferrin preparations (native or oxidized) in the incubation medium had little effect on the rate of iron incorporation into heme from diferric chicken ovotransferrin. The effect of modifying the periodate-susceptible methionine residues in chicken ovotransferrin was small but significant. These methionine residues do not appear critical for the interaction of chicken ovotransferrin with the chicken embryo red blood cell receptors, the incorporation of chicken ovotransferrin into the cell, or the release of iron from chicken ovotransferrin for heme synthesis. PMID- 2982409 TI - Effects of dithiothreitol, dithioerythritol and chelating agents on 5' nucleotidase from bull seminal plasma. AB - 5'-Nucleotidase from bull seminal plasma is inhibited by dithiothreitol and dithioerythritol. These reactives proved to dissociate the dimeric glycoprotein 5'-nucleotidase of Mr 160 000 into two subunits of apparent Mr 80 000, indicating that the subunits are held together by interchain disulfide bridges. HPLC determinations of cysteic acid and carboxymethylcysteine protein derivatives resulted in 50 +/- 3 half-cystine plus cysteine residues, while 1.9 +/- 0.4 free cysteine residues were estimated by HPLC analysis. The enzyme is inhibited by EDTA and EGTA, and the inhibition appears to be of the non-competitive type for both the chelating agents. Experiments for the enzyme activity recovery by MgCl2 and CaCl2 additions, after the EDTA and EGTA treatments in the presence of 8 M urea, are reported. PMID- 2982410 TI - Purification and mode of action of a microsomal endoribonuclease from rat liver. AB - An endoribonuclease has been purified nearly to homogeneity from rat liver microsomes, and its mode of action and general properties were studied. The enzyme had an apparent molecular weight of 58 000, as estimated by both gel filtration and SDS-polyacrylamide gel electrophoresis and produced oligonucleotides from poly(A), poly(U) and poly(C). No mononucleotide was obtained by the enzymatic hydrolysis of the above substrates. The enzyme made endonucleolytic cleavages which generated 5'-phosphate-terminated oligonucleotides. It was suggested that the existence of at least (Ado5'P)2 residues at both sides of the cleavage bond was necessary for the action of the endoribonuclease. Divalent cations (Mg2+ or Mn2+) were required for the enzymatic activity, while K+ inhibited the enzyme. Spermine stimulated the enzymatic activity in the presence of 1 mM Mg2+. PMID- 2982411 TI - Purification and mode of action of a microsomal exoribonuclease from rat liver. AB - An exoribonuclease has been purified nearly to homogeneity from rat liver microsomes and its mode of action and general properties were studied. The molecular weight values for the enzyme, as estimated by gel filtration and SDS polyacrylamide gel electrophoresis, were 88 000 and 92 000, respectively. The enzyme produced, via a processive mechanism Ado5'P as the only product from poly(A). The results of the hydrolysis of 4 S (Ado5'P)n and (Ado3'P)n by the exoribonuclease with or without alkaline phosphatase and the inhibition of the enzymatic activity by oligonucleotides having a 3'-phosphate group in the 3' terminus suggested that the degradation proceeds in the 3' to 5' direction. These findings were confirmed by the analysis of hydrolyzed products of various oligoadenylates and Ado3'PUrdPGuo and by the comparison of the rates of hydrolysis of (Ado3'P)2Ado by the enzyme in the presence of varying amounts of (Ado3'P)3. Mg2+ was required for the enzymatic activity, and Mn2+ partially substituted for Mg2+. The activity of the enzyme was stimulated by K+ and spermine. PMID- 2982412 TI - Proteinase inhibitory activity released from the horseshoe crab blood cell during exocytosis. AB - The blood cell of the horseshoe crab, Limulus, is packed with granules that can be stimulated to release their contents by exocytosis. We have identified a family of proteinase inhibitors in the released materials. Included is a factor similar to the alpha 2-macroglobulin homologue present in the plasma and acid stable and acid-instable active-site inhibitors. The acid-stable factor is active against both serine (trypsin, chymotrypsin) and metal (thermolysin) proteinases. The trypsin- and chymotrypsin-inhibitory activity has a molecular weight of 6100, as determined by gel-filtration chromatography. PMID- 2982413 TI - Do vanadate polyanions inhibit phosphotransferase enzymes? AB - Decavanadate inhibits hexokinase, adenylate kinase and phosphofructokinase; neither mono-, tri nor tetrameric vanadate anion is an inhibitor. Decavanadate inhibits phosphofructokinase obtained from bacterial and protistic sources. No form of vanadium(V) anion inhibits galacto-, glycero-, pyruvate and creatine kinase, or inorganic pyrophosphatase. Decavanadate appears to be a non competitive inhibitor of both hexokinase substrates. PMID- 2982414 TI - Raman characterization of human leukocyte myeloperoxidase and bovine spleen green haemoprotein. Insight into chromophore structure and evidence that the chromophores of myeloperoxidase are equivalent. AB - Soret excitation resonance Raman spectroscopy has been used to characterize dimeric human leukocyte myeloperoxidase (donor:hydrogen peroxide oxidoreductase, EC 1.11.1.7) and monomeric bovine spleen green haemoprotein. The spectra of the two proteins, under the same conditions of iron valence and ligation, are essentially identical. Owing to strong symmetry reduction effects, the spectra are more complex than usually observed for haemoproteins. It is possible, however, to assign the high-frequency vibrations and, from these assignments, to determine structural features of the iron chromophores. In the resting protein, the iron adopts a six-coordinate high-spin configuration in both proteins; cyanide addition produces six-coordinate low-spin species, and in the ferrous enzymes the iron appears to be five-coordinate and high-spin. The proteins are stable to laser excitation and do not photoreduce under illumination. No evidence is found for unusual peripheral substituents, such as formyl or protonated Schiff's base group, in conjugation with the main chromophore in the native protein. The vibrational data are consistent with an iron chlorin chromophore, although other electronic effects, in addition to those produced by porphyrin ring reduction, are necessary to account for the optical properties of the proteins. The similarity in Raman spectra for myeloperoxidase and green haemoprotein indicates that the two iron sites in myeloperoxidase are equivalent. PMID- 2982415 TI - Purification of a high-molecular-weight inhibitor of the calcium-activated proteinase. AB - An inhibitor of the muscle calcium-activated proteinases has been purified from porcine skeletal muscle by using DEAE-cellulose column chromatography, thermal treatment, Sephacryl S-400 column chromatography in 6 M urea and Sephacryl S-300 column chromatography in 6 M urea. Sodium dodecyl sulfate polyacrylamide slab gel electrophoresis shows that the purified inhibitor is homogeneous and has a subunit molecular weight of 172 000. The inhibitor inactivates both the low- and high-calcium-requiring forms of the calcium-activated proteinase but does not inhibit other proteinases against which it has been tried. It thus appears that the inhibitor is specific for the calcium-activated proteinase. Studies using homogeneous inhibitor and high-calcium-requiring proteinase show that one molecule of the inhibitor can inactivate up to eight molecules of the calcium activated proteinase. Inactivation of the calcium-activated proteinase by the inhibitor cannot be reversed by calcium concentrations as high as 25 mM, thus eliminating the possibility that the inhibitor functions by chelating calcium. The inhibitory peptide appears to be extremely susceptible to proteolysis during its isolation. Even in the presence of synthetic proteinase inhibitors different inhibitor preparations yield homogeneous inhibitory peptides ranging in molecular weight from 145 000 to 172 000. Preparative electrophoresis and column chromatography have been used to isolate putative proteolytic breakdown products of the 172 kDa peptide at 145, 114, 41 and 29 kDa. PMID- 2982416 TI - Effect of alcohols on arachidonic acid metabolism in murine mastocytoma cells and human polymorphonuclear leukocytes. AB - The effects of alcohols on the formation of leukotrienes, 5-HETE and prostaglandin D2 in mastocytoma cells and human neutrophils were studied. In murine mastocytoma cells, alcohols appear to have at least two different effects on the production of these arachidonic acid metabolites. At low levels of cellular arachidonic acid achieved after stimulation with calcium ionophore A23187 or addition of low levels of exogenous arachidonic acid, alcohols appear to have a general inhibitory effect on the production of lipoxygenase metabolites. In the presence of higher concentrations of cellular arachidonic acid, ethanol and methanol stimulated the production of lipoxygenase metabolites, but had no large stimulatory effect on the cyclo-oxygenase metabolite, prostaglandin D2. Under these conditions, n-propanol and t-butanol have inhibitory effects on leukotriene production. Human neutrophils are less sensitive to ethanol than mastocytoma cells, but stimulatory effects were still found at high ethanol concentrations (220-430 mM). PMID- 2982417 TI - The stimulation and binding of CTP: phosphorylcholine cytidylyltransferase by phosphatidylcholine-oleic acid vesicles. AB - The activity of the low molecular weight form of cytidylyltransferase from fetal lung cytosol and adult liver cytosol was stimulated more by phosphatidylcholine oleic acid (1:1 molar ratio) vesicles than by phosphatidylglycerol vesicles. Phosphatidylcholine alone did not stimulate the activity, while oleic acid alone produced only slight stimulation. Vesicles prepared from phosphatidylinositol, phosphatidylglycerol-cholesterol (2:1) and phosphatidylglycerol phosphatidylcholine (1:1) all stimulated the activity to the same extent. Phosphatidylcholine-oleic acid vesicles (molar ratio 2:1) produced less stimulation than 1:1 vesicles. Phosphatidylcholine-palmitic acid vesicles (2:1) were about 50% as active as the corresponding phosphatidylcholine-oleic acid vesicles. All vesicles were in the size range of small unilamellar vesicles as judged by Sephacryl S-1000 chromatography. Stimulation also occurred when phosphatidylcholine vesicles and oleic acid were added separately to the assay. The stimulation by phospholipid vesicles was correlated with the ability of the vesicles to bind cytidylyltransferase, determined by sucrose density centrifugation of the enzyme-vesicles mixtures. We conclude that the stimulation of soluble cytidylyltransferase occurs through binding of the enzyme to anionic membrane surfaces. Suitable anionic membranes can be prepared either from anionic phospholipids, or by the addition of anionic lipids (unesterified fatty acids or phosphatidylglycerol) to phosphatidylcholine. PMID- 2982418 TI - Regulation of phospholipid synthesis by intracellular phospholipases in fetal rabbit type II pneumocytes. AB - Exposure of fetal type II pneumocytes to phospholipase A2 inhibitors led to significantly reduced choline uptake and decreased synthesis of total and disaturated phosphatidylcholines from both [methyl-14C]choline and [9,10(n) 3H]palmitate precursors. The percentage of the total synthesized phosphatidylcholine recovered as disaturated phosphatidylcholine was increased when compared to that in control cultures, suggesting that unsaturated phosphatidylcholine synthesis was reduced to a greater extent than that of the disaturated species. Synthesis of sphingomyelin and phosphatidylethanolamine from labeled palmitate was also reduced, whereas that of phosphatidylinositol and phosphatidylglycerol was significantly increased. Addition of phospholipase C resulted in increased synthesis of phosphatidylcholine from both labeled precursors; no significant changes were found in synthesis of most of the other 3H-labeled lipids. Added phospholipase A2 did not lead to any changes in either choline or palmitate incorporation. However, when melittin (a phospholipase A2 activator) was added to the cultures, greater incorporation of both palmitate and choline was observed, along with a significant increase in the percentage of total cellular radioactivity in 14C-labeled lipids, indicating also stimulation of phosphatidylcholine synthesis. A marked increase in CTP: phosphorylcholine cytidylyltransferase activity was found after treatment of the cultures with phospholipase C. Exposure to quinacrine also increased the activity of this enzyme. Addition of phospholipase C and melittin to prelabeled pneumocyte cultures accelerated degradation of cell phospholipids and the release of free fatty acids as the main degradation products. These findings suggest that intracellular phospholipases are regulators of synthesis of surfactant phospholipids in fetal type II pneumocytes, and that activation or inhibition of these phospholipases could represent a mechanism through which hormones and pharmacological agents modify surfactant and other phospholipid synthesis. PMID- 2982419 TI - Influence of restricted food intake on brown adipose tissue function in genetically obese mice (genotype, ob/ob). AB - Measurements were made of cytochrome c oxidase activity and the GDP-binding capacity of mitochondria in brown adipose tissue of genetically obese mice and wild-type siblings, to estimate the thermogenic capacity of the tissue. The binding capacity was decreased in ad libitum fed obese animals compared with wild type animals. Limited feeding of obese animals to restrict their body weight caused a large increase in the binding capacity of the tissue, which was greater than that in wild-type animals fed either ad limitum or on a limited diet. The decreased binding capacity of brown adipose tissue mitochondria in obese mice appears to be a consequence of ad libitum feeding and therefore not a cause of the obesity. Limit feeding of obese animals also corrected their characteristic hypothermia at low ambient temperature. The large increase in the thermogenic capacity of brown adipose tissue in obese animals, induced by limited feeding, may account for the vital improvement of their thermoregulation. However, close similarities were found between obesity hypothermia and hypothermia induced in wild-type animals by restraint. It is suggested that changes in posture caused by obesity, resulting in increased loss of body heat, may be important in the development of obesity hypothermia. Obese animals fed less than wild-type grained more weight than wild-type animals, indicating that the high thermogenic capacity of their brown adipose tissue did not function to regulate their calorie intake. PMID- 2982420 TI - Involvement of superoxide radicals on adrenochrome formation stimulated by arachidonic acid in bovine heart sarcolemmal vesicles. AB - Highly purified sarcolemmal membranes prepared from bovine heart muscle produced superoxide radicals, especially when incubated with NADPH or NADH, as revealed by the oxidation of adrenaline to adrenochrome. The reaction was inhibited by superoxide dismutase or by heat denaturation of the sarcolemmal vesicles. Less evident was the inhibitory effect shown by catalase, while mannitol, deferoxamine or dicumarol were uneffective. The formation of adrenochrome was an oxygen dependent reaction with a Km for adrenaline of 8-10 microM. Moreover, the reaction was inhibited by preincubating the sarcolemmal membranes with propranolol, while the alpha-antagonist phentolamine was without effect. Adrenaline oxidation was unaffected by the presence of exogenous linolenic acid or methylarachidonic acid, while arachidonic acid, with a Km for this reaction of 175 microM, showed a marked stimulatory effect. This activation was suppressed by superoxide dismutase, catalase and NaCN, while mannitol was without effect. Moreover, the reaction was blocked by the cyclooxygenase inhibitor indomethacin, differently from the lipooxygenase inhibitor nordihydroguaiaretic acid. Also, the incubation of the sarcolemmal vesicles with phospholipase A2 and calcium produced a stimulation of adrenochrome formation which was partially suppressed by albumin. In the experiments using arachidonic acid or phospholipase A2, the addition of indomethacin blocked the adrenaline oxidation. These results indicate that arachidonic acid accentuated the heart sarcolemmal adrenochrome formation presumably by participating in the cyclooxygenase reaction. PMID- 2982421 TI - Synthesis of transcobalamin II by cultured human hepatocytes. AB - Cultured HepG2 cells, derived from a human hepatoma synthesized and released unsaturated, immunoreactive transcobalamin II. Synthesis was confirmed by the blocking with inhibitors of protein synthesis and by incorporation of tritiated leucine into transcobalamin II. PMID- 2982422 TI - A calmodulin-dependent protein kinase in Rous sarcoma virus-transformed rat cells and normal liver. AB - A calmodulin-dependent protein kinase has been purified extensively from a Rous sarcoma virus-transformed rat cell line (RR1022) and from normal rat liver. The calmodulin-dependent protein kinase activity was manifested by in vitro phosphorylation of a single Mr 57 000 endogenous phosphoprotein (pp57) present in both the virally transformed cells and normal rat liver. The calmodulin-dependent protein kinase from transformed cells fractionated with the viral src gene product, pp60v-src, through a 650-fold purification of the oncogene product. However, purification of the calmodulin-dependent protein kinase from normal liver demonstrated that the calmodulin-dependent kinase was distinct from pp60v src. Phosphorylation of pp57 by the kinase purified from the transformed cell line required Ca2+ and calmodulin, was inhibited by EDTA and was unaffected by cAMP or the heat- and acid-stable protein inhibitor of cAMP-dependent protein kinase. Troponin C did not substitute for calmodulin. A virtually identical calmodulin-dependent protein kinase activity was purified from rat liver by affinity chromatography on calmodulin-Sepharose. Phosphorylation of pp57 by the affinity-purified liver protein kinase was also observed, and required Ca2+ and calmodulin. EGTA and trifluoroperazine inhibited pp57 phosphorylation. The calmodulin-dependent protein kinase reported here did not phosphorylate substrates of known calmodulin-dependent protein kinases in vitro (myosin light chain, phosphorylase b, glycogen synthase, microtubule-associated proteins, tubulin, alpha-casein). Because none of these proteins served as substrates in vitro and pp57 was the only endogenous substrate found, the properties of this enzyme appear to be different from any previously described calmodulin-dependent protein kinase. PMID- 2982423 TI - Regulation of Ca2+ transport by cyclic 3',5'-AMP-dependent and calcium-calmodulin dependent phosphorylation of cardiac sarcoplasmic reticulum. AB - Canine cardiac sarcoplasmic reticulum is phosphorylated by cyclic AMP-dependent and by Ca2+-calmodulin-dependent protein kinases on a 22 kDa protein, called phospholamban. Both types of phosphorylation have been shown to stimulate the initial rates of Ca2+ transport. To establish the interrelationship of the cAMP dependent and Ca2+-calmodulin-dependent phosphorylation on Ca2+ transport, cardiac sarcoplasmic reticulum vesicles were preincubated under optimum conditions for: (a) cAMP-dependent phosphorylation, (b) Ca2+-calmodulin-dependent phosphorylation, and (c) combined cAMP-dependent and Ca2+-calmodulin-dependent phosphorylation. Control vesicles were treated under identical conditions, but in the absence of ATP, to avoid phosphorylation. Control and phosphorylated sarcoplasmic reticulum vesicles were subsequently centrifuged and assayed for Ca2+ transport in the presence of 2.5 mM Tris-oxalate. Our results indicate that cAMP-dependent and Ca2+-calmodulin-dependent phosphorylation can each stimulate calcium transport in an independent manner and when both are operating, they appear to have an additive effect. Stimulation of Ca2+ transport was associated with a statistically significant increase in the apparent affinity for calcium by each type of phosphorylation. The degree of stimulation of the calcium affinity was relatively proportional to the degree of phospholamban phosphorylation. These findings suggest the presence of a dual control system which may operate in independent and combined manners for regulating cardiac sarcoplasmic reticulum function. PMID- 2982424 TI - Synthesis and secretion of protein C inhibitor by the human hepatoma-derived cell line, Hep G2. AB - The site of synthesis of protein C inhibitor, a recently identified human plasma inhibitor against activated protein C, is not known. We have studied the production and secretion of protein C inhibitor by an established human liver cell line derived from hepatocellular carcinoma (Hep G2). The concentration of protein C inhibitor, as measured by a specific radioimmunoassay, increased in the medium of Hep G2 cells with time. There was no evidence for a significant intracellular pool of this protein. Protein C inhibitor secreted from Hep G2 cells (G2 protein C inhibitor) inhibited the activity of purified activated protein C in a functional assay. De novo synthesis of protein C inhibitor was demonstrated by the presence of specific immunoprecipitable radioactivity in the medium after 5 h of labeling of the cells with [35S]methionine. Analysis of the immunoprecipitates by SDS-polyacrylamide gel electrophoresis showed a peak of radioactivity corresponding to Mr 57 000. These results indicate that the liver is a site of protein C inhibitor production. PMID- 2982425 TI - Activation of guinea-pig and bovine neutrophil NADPH oxidase by N,N' dicyclohexylcarbodiimide. AB - Dicyclohexylcarbodiimide (DCCD) is a potent stimulant of superoxide generation in guinea-pig peritoneal and bovine blood neutrophils. The dependence of DCCD elicited respiratory burst on the composition of the medium was investigated. At 37 degrees C, the superoxide generation was short-lived and a rapid losses of enzymatic activity was observed; at 0 degree C, the activity could be preserved for hours. Superoxide generation by whole cells was accompanied by exocytic degranulation. Prolonged incubation with DCCD at 37 degrees C resulted also in a progressive loss of cellular integrity evidenced by the release of a fraction of lactate dehydrogenase. Km values of the particulate NADPH oxidase isolated from DCCD-triggered guinea-pig and bovine cells were 31.7 and 50.0 microM, respectively. Cells pre-equilibrated with the potential sensitive dye Di-S-C3-(5) exhibited changes in the transmembrane potential upon stimulation. Stimulation with DCCD resulted also in the release of membrane-associated calcium, indicated by quenching of the fluorescence of chlortetracycline-loaded neutrophils. Both effects were observed also in human neutrophils which did not generate superoxide upon exposure to DCCD. The mechanism of DCCD-induced responses is discussed. PMID- 2982426 TI - The coupling of metabolic to secretory events in pancreatic islets. The possible role of glutathione reductase. AB - The participation of glutathione reductase in the process of nutrient-stimulated insulin release was investigated in rat pancreatic islets exposed to 1,3-bis(2 chloroethyl)-1-nitrosourea (BCNU). BCNU caused a time-and dose-related, irreversible inhibition of glutathione reductase activity. This coincided with a fall in both GSH/GSSG ratio and the thiol content of the islets. Pretreatment of the islets with BCNU inhibited the oxidation of glucose and its stimulant action upon both 45Ca net uptake and insulin release. Although BCNU (up to 0.5 mM) failed to affect the oxidation of L-leucine and L-glutamine, it also caused a dose-related inhibition of insulin release evoked by the combination of these two amino acids. The latter inhibition was apparently not fully accounted for by the modest to negligible effects of BCNU upon 45Ca uptake, 45Ca efflux, 86Rb efflux and cyclic AMP production. Since BCNU failed to inhibit insulin release evoked by the association of Ba2+ and theophylline, these results support the view that glutathione reductase participates in the coupling of metabolic to secretory events in the process of nutrient-stimulated insulin release. However, the precise modality of such a participation, for example the control of intracellular Ca2+ distribution, remains to be elucidated. PMID- 2982427 TI - The characterization and regulation of a polyamine-responsive, cyclic nucleotide independent protein kinase activity in the mouse mammary gland. AB - The existence of two cyclic nucleotide-independent protein kinases in the cytosolic extract of mouse mammary gland has been determined via DEAE-cellulose and Sephacryl column chromatography. Both enzymes phosphorylated casein in the absence of the exogenous cyclic nucleotides, cAMP and cGMP. One protein kinase was found to have a molecular weight of approx. 30 000, while the other was found to have a molecular weight in the range 150 000-250 000. The activity of the larger species was enhanced by polyamines and inhibited by heparin. This enzyme utilized both ATP and GTP as phosphate donors; the apparent Km values were 10 and 16 microM, respectively. The lower molecular weight protein kinase was not affected by either polyamines or heparin and utilized only ATP (Km = 8 microM) as the phosphate donor. The polyamine-responsive protein kinase activity in the mammary gland varied as a function of the reproductive development of the mouse. The activity was relatively low in the virgin and primiparous stages, increased during pregnancy and peaked during lactation. Studies using mammary organ culture indicated that the combination of insulin (5 micrograms/ml), cortisol (1 micrograms/ml) and prolactin (5 micrograms/ml) maintained the polyamine responsive protein kinase activity that was present in noncultured tissue. In the absence of prolactin, however, the kinase activity was significantly lower than that observed in the three-hormone system. When dibutyryl cyclic AMP (0.5 mM) was added to the medium along with the three hormones, a significant decrease in enzyme activity was found. Slab gel electrophoresis and autoradiography showed that the majority of the phosphorylated endogenous substrates in the cytosolic fraction were caseins. The results of this study suggest that the polyamine responsive protein kinase may play an important role in the growth and development of the mammary gland. PMID- 2982428 TI - Stimulation of skeletal-derived cell cultures by different electric field intensities is cell-specific. AB - Pulsed electric stimulation, coupled capacitively to different cell cultures of skeletal origin, caused immediate changes in the cellular levels of cyclic AMP and a later enhanced DNA synthesis. Changes both in cyclic AMP level and DNA synthesis were correlated with the strength of the applied electric field. Cultures of calvaria bone cells which contain mainly two cell types, parathyroid hormone responsive cells (osteoblast-like) and prostaglandin E2 responsive cells (fibroblast-like), respond to both low (13 V/cm) and to high (54 V/cm) electric field strength, with no response at intermediate (24 V/cm) field strength. Rat epiphyseal cartilage responded like bone cells both to low and high field intensities, while rat condylar cartilage responded only to the intermediate field strength. Moreover, subcultures of calvaria bone cells, which lost their osteoblastic phenotype expression during subculturing, were responsive only to low field strength. On the other hand, osteoblast-enriched cultures, derived from calvaria bone grown in low calcium, were responsive only to the high field strength. These findings suggest that the response to various electric field intensities is cell-specific and might be used as an additional parameter to characterize cell types. Our study points to the possibility that when exposing a whole organ to an electrical stimulation it is possible to affect specifically only one cell population out of the many cell types existing in the organ. PMID- 2982429 TI - Differentiation-associated increase of cAMP-dependent type II protein kinase in a murine preadipose cell line (ST 13). AB - The activity of cAMP-dependent protein kinase and cAMP binding activity were studied during the differentiation of ST 13 murine preadipocytes into adipocytes. We found that both activities were marginally detectable in preadipose cells and increased remarkably when the cells were induced to differentiate, preceding by several days the morphological adipose conversion. The increased cAMP-dependent protein kinase was identified as type II enzyme by means of DEAE-Sephacel chromatography and by photoaffinity labeling with 8-azido[3H]cAMP. We further showed that the increase of protein kinase activity was specific to cell differentiation with the aid of modulators of the adipose conversion (insulin, fetal bovine serum, retinoic acid and 5-bromodeoxy-uridine). We propose that the increased expression of type II cAMP-dependent protein kinase would be a biochemical index of differentiation in ST 13 preadipocytes. PMID- 2982430 TI - A comparison of the spin labels MAL-3 and TEMPAMINE for measuring the internal microviscosity of human erythrocytes. AB - In a recent paper, Daveloose et al. (Daveloose, D., Fabre, G., Berleur, F., Testylier, G. and Letterrier, F. (1983) Biochim. Biophys. Acta 763, 41-49) described a technique to measure the internal microviscosity of erythrocytes using the spin label MAL-3 (2,2,5,5-tetramethyl-3-malimidopyrrolidinyl-N-oxyl) to supercede the use of TEMPAMINE (2,2,6,6-tetramethylpiperidine-N-oxyl-4 amine)ferricyanide. They cite the long time (12 h) required for TEMPAMINE to enter the erythrocyte interior and the 'unphysiological' nature of the potassium ferricyanide required to isolate the TEMPAMINE signal inside the erythrocytes as reasons for developing this technique. In the present work, the penetration of TEMPAMINE into the erythrocyte interior is found to be pH-dependent and on the order of seconds, not hours. The slow penetration of TEMPAMINE described by Daveloose et al. probably is a result of their using trimethylTEMPAMINE which is membrane-impermeable. High concentrations of ferricyanide do affect red cell morphology in a reproducible fashion. The internal microviscosity as measured by TEMPAMINE reflects this. MAL-3 does not. Sample preparation time using TEMPAMINE ferricyanide is about 1 min compared to about 40 min for the MAL-3 technique. In the presence of ferricyanide or ascorbate, MAL-3-labeled cells show slow loss of signal intensity which indicates that MAL-3 leaks out of the cells. Both the TEMPAMINE-ferricyanide and MAL-3 techniques have advantages and disadvantages. PMID- 2982431 TI - Phosphorylation of lens membranes with a cyclic AMP-dependent protein kinase purified from the bovine lens. AB - We report the phosphorylation of lens membranes with a cAMP-dependent protein kinase isolated from bovine lenses. The holoenzyme was eluted from DEAE agarose at less than 100 mM NaCl and from gel filtration columns with a relative molecular weight of 180 000. The regulatory subunit was identified with the affinity label 8-azido-[32P]cAMP. Four focusing variants with relative molecular weights of 49 000 were seen on two-dimensional gels. The catalytic subunit was purified approx. 5000-fold and migrated at 42 000 Mr on SDS gels. Based on these observations, the enzyme is classified as a Type I cAMP-dependent protein kinase. Purified lens plasma membranes were incubated with the holoenzyme or its catalytic subunit in the presence of 32P-labeled ATP. Several membrane proteins, including the major lens membrane polypeptide, MP26, were shown to be substrates for the kinase in this reaction. MP26 appears to be the major component of intercellular junctions in the lens. Studies with protease treatments on labeled membranes appeared to localize the phosphorylation sites to the cytoplasmic side of the membrane. PMID- 2982432 TI - A study of rat epididymal sperm adenosine 3',5'-monophosphate-dependent protein kinases: maturation differences and cellular location. AB - The photoaffinity analog [32P]8-N3 cAMP (8-azido adenosine 3',5'-monophosphate was used to analyze the membrane sidedness of rat sperm cAMP binding proteins during epididymal maturation. Evidence is presented here which supports the hypothesis that 35-45% of the regulatory subunits of the Type I and Type II cAMP dependent protein kinases are readily available to externally added cyclic nucleotide. It was observed by sodium dodecyl sulfate gel electrophoresis (SDS PAGE) and autoradiography that only two rat sperm proteins (Mr = 49K and 55K) were photolabeled which comigrated on gels with partially purified Type I and Type II regulatory subunits, respectively. Both of these photolabeled epididymal sperm proteins were saturated at physiological titers of [32P]8-N3cAMP and photoincorporation of [32P]8-N3 cAMP was specific since other SDS-resolvable sperm proteins did not photoincorporate the analog. Caput and cauda sperm protein photoincorporation could be effectively blocked by low levels of cAMP, but not by cGMP, ATP or GTP. Sperm epididymal maturation coincided with changes in the cAMP dependent protein kinase subunits since cauda sperm contained more available Type II than did caput sperm. A subcellular analysis of cAMP-dependent protein kinase regulatory subunit in head and tail fractions was done for caput and cauda sperm and demonstrated that the tail fractions showed more photo-labeling of both Type I and II regulatory subunits than did the head fractions. PMID- 2982433 TI - An enzymatic method for dissociation of intact follicles from the hamster ovary: histological and quantitative aspects. AB - An enzymatic method was developed to collect intact follicles at different stages of development from cyclic hamsters to study ovarian folliculogenesis under various circumstances. Ovaries from 6 adult hamsters on each day of the cycle (Day 1 = ovulation) were collected, corpora lutea and large preantral and antral follicles were dissected, and follicles saved. Minced ovaries were then incubated with a mixture of collagenase, DNAse and pronase at 37 degrees C for 20 min to disperse intact follicles. Histological studies with 2191 isolated follicles revealed 10 different stages of follicular development (depending on the number of granulosa cell layers surrounding the oocyte and development of the antrum). Of the total follicular population, 14% showed signs of atresia, with 50% of those having 1-3 layers of granulosa cells (Stages 1-3); a second peak of 18% was observed in antral follicles (Stages 8-10). No signs of thecal cells were evident until the follicles reached Stage 6 (7-8 layers of granulosa cells), which possibly accounts for reduced atresia in this class and beyond. Ultrastructural study revealed that there were no signs of morphological damage to the basement membrane or to other subcellular organelles in the small preantral follicles. The presence of subnuclear lipid droplets in follicles with 3 layers of granulosa cells provided evidence for potential steroidogenesis by small follicles. The number of Stage 1-10 follicles was remarkably constant throughout the estrous cycle (460 +/- 34 per animal on Day 1 vs. 492 +/- 66 on Day 4). The usefulness of this method in analyzing follicular kinetics is illustrated in experiments involving hypophysectomy and the effects of unilateral ovariectomy. This procedure offers an improved method to study the factors responsible for the growth and the differentiation of small preantral follicles in the mammalian ovary. PMID- 2982434 TI - Potential of a quantal response as a mechanism for oscillatory behavior: implications for our concepts of hormonal control mechanisms. AB - This article describes the potential of a quantal (i.e., all-or-none) response as a model for understanding the interactions between endocrine, paracrine and autocrine hormones. We review the general features of continuous and discontinuous (i.e., oscillating) quantal models including the role of a threshold. In addition, we also describe a few of the many different biochemical mechanisms which may give rise to quantal behavior. One of the more attractive schemes involves the coordinate regulation of opposing biochemical pathways resulting from phosphorylation of hormone receptors and/or rate-limiting enzymes. At least one hormone receptor (i.e., that for insulin) and many rate-limiting enzymes which control the flow of metabolites through a variety of metabolic pathways can be phosphorylated at multiple sites by one or more protein kinases. Phosphorylation may enhance or inhibit the activities of these proteins depending on which sites are modified. Furthermore, since phosphorylation of some sites on a protein may enhance the ability of phosphoprotein phosphatases to dephosphorylate other sites responsible for biological activity of the protein, phosphorylation also has the potential to produce a discontinuous quantal response. Quantal response mechanisms may alter our notions of endocrine regulation. When a quantal response mechanism is applied to a simple negative feedback model similar to that which was originally postulated to explain the interactions between gonadotropin and steroid hormonal levels, the model can account for the oscillations in hormone levels even when the input is constant. Conversely, when a graded mechanism is applied to the same negative feedback model, the model will almost certainly result in constant hormone levels. Further, the model illustrates that small changes in rate constants and thresholds of response, amplification of hormonal signals, and degradation of intermediate regulators can produce large shifts in the output of the system. These may account for the variability in hormonal levels observed in some endocrine systems. Finally, the high sensitivity of the quantal response mechanism accounts for the data which suggest that gonadotropins may play permissive rather than causal roles in regulation of gonadal function. Since increasing evidence suggests that all cells of a given type may not be equal in terms of hormonal responsiveness, measurements of response in single cells over short time periods will be needed before the role of a quantal response can be determined and endocrine regulation will be fully understood. PMID- 2982435 TI - Setting properties of bone cement with added synthetic hydroxyapatite. AB - Synthetic hydroxyapatite powder was added to commercial bone cement to improve its physico-mechanical properties. The study of the setting properties showed the considerable effect of hydroxyapatite (HA) on the polymerization process; in particular statistical treatment of the experimental results revealed a significant relative minimum of the exothermic peak value at HA 5% weight percentage--an effect attributed to the HA 'crumbling' action on the microporosity features. The relationship between the interface (air and monomer bubbles/acrylic resin) versus dough composition was evaluated: the comparison between this last one and the corresponding internal energy variation supports the above hypothesis. PMID- 2982436 TI - [ATPase activity of the heart microsomes, the regulation of calcium transport in the microsomes and the calmodulin content in experimental myocardial infarct]. AB - Calmodulin and cAMP were demonstrated to have no stimulating effect on Ca2+ transport in the sarcoplasmic reticulum of the dog heart in experimental myocardial infarction as compared to that in the uninvolved myocardium (control). Introduction to the incubation medium of exogenous protein kinase in addition to calmodulin and cAMP provoked an approximately 35% increase in 45Ca accumulation in microsomes of the impaired myocardium as compared with the system containing no exogenous protein kinase. Under the same conditions, the control showed a 75% increase in 45Ca accumulation. A reduction in the activity of Ca2+-activated ATPase of the reticulum and translocation of calmodulin activity from the membrane fraction of cardiomyocytes to cytosol were recorded in myocardial infarction. PMID- 2982437 TI - [Conditions for initial chromatin endonucleolysis and the characteristics of its products]. AB - The restricted initial autohydrolysis of chromatin is used for obtaining its matrix-active fractions. Application of the method for isolating cell nuclei in the presence of 10 mM CaCl2 makes it possible to appreciably inhibit the cleavage of chromatin DNA in the course of isolation, namely to 0.9% according to the acid soluble fraction. It is shown that at the initial stages of autohydrolysis the method of solubilization is by one order of magnitude more sensitive as compared to the measurement according to the acid-soluble fraction. Initial autohydrolysis is characterized by rapid increment of the amount of the solubilized fraction, delayed formation of mononucleosomes and by the capacity to a 20% activation in the presence of 10(-5) M histamine. PMID- 2982438 TI - Decreased ecto-5'nucleotidase activity of peripheral blood lymphocytes in human monoclonal gammopathies: correlation with tumor cell kinetics. AB - Ecto-5'nucleotidase (5'NT) activity of peripheral blood (PB) lymphocytes was determined in 31 patients with serum monoclonal gammopathies (MG). Twenty-one patients had a diagnosis of multiple myeloma (MM), and ten patients had monoclonal gammopathy of undetermined significance (MGUS). The proliferative activity of the bone marrow plasma cells (LI%) was investigated in 28 of these MG patients by means of tritiated thymidine uptake evaluated by simultaneous autoradiography and cytoplasmic immunofluorescence. 5'NT activity was significantly lower in MG patients as compared with normal controls. MM patients had lower 5'NT activity than MGUS patients, but the difference was not significant. By contrast, MM had significantly higher LI% than MGUS patients. There was a linear regression of 5'NT on LI% which was statistically significant: the higher the LI%, the lower the 5'NT. Because the LI% is an accurate prognostic and monitoring factor in MG, this correlation indicates that 5'NT may be of assistance in predicting the clinical progress of MG patients. In seven MGs, the PB T and B lymphocytes were studied separately. The T cell subpopulation was 5'NT deficient compared to the normal controls, shown as a significant linear regression of T cell 5'NT on the LI%. This suggests that in MG there may be an alteration of nonneoplastic T lymphocytes correlated with tumor growth. The OKT8+ lymphocytes were mainly responsible for the 5'NT deficiency of unseparated T lymphocytes. PMID- 2982439 TI - Epstein-Barr virus genomes are restricted to secondary neoplastic cells following bone marrow transplantation. AB - DNA from mononuclear blood and tumor cells from 33 patients undergoing bone marrow transplantation for leukemia was examined for the presence of Epstein-Barr virus (EBV) genomes by blot hybridization. Four groups of patients were studied soon after engraftment, during long-term remission, after relapse of the original leukemia, and after development of secondary B cell neoplasms. Only the cells of patients with secondary neoplasms demonstrated EBV genomes, where all five adequately studied samples were positive. Samples from all other patient categories were negative for EBV genomes. We conclude that EBV genomes do not frequently persist in normal engrafted lymphocytes or in mononuclear cells of patients suffering recurrent leukemia. These results are consistent with EBV playing a role in the genesis of secondary B cell neoplasms following bone marrow transplantation. PMID- 2982440 TI - Effects on the buoyant density of rabbit platelets of ADP and agents that increase the concentration of cyclic AMP. AB - Rabbit platelets were aggregated by adenosine diphosphate (ADP), allowed to deaggregate and then separated into density subpopulations by centrifugation through discontinuous Stractan density gradients. Although ADP causes little or no release of the contents of the amine storage granules of rabbit platelets, ADP caused a decrease in platelet density as compared with control platelets subjected to the same procedures except for exposure to ADP. The density change persisted for at least four hours. The apparent size of platelets stimulated with ADP increased initially, but returned to control values during a one-hour period. A similar decrease in platelet density was observed with an albumin density gradient. Under conditions in which aggregation did not occur in response to ADP with ethylenediaminetetraacetic acid (EDTA) in the medium, little or no decrease in platelet density was observed. Agglutination with polylysine did not change platelet density. Thus, not only agents such as thrombin and plasmin that cause the release of the contents of the platelet granules decrease platelet density, but ADP also has this effect. Platelets would be exposed to all of these stimuli during thromboembolic processes, and their effect on platelets may account for the decrease in platelet density observed previously in experiments with rabbits with indwelling aortic catheters. Agents that increase the concentration of cyclic AMP (cAMP) in platelets (PGE1, adenosine, dibutyryl cAMP, forskolin, and papaverine) also decreased platelet density. This effect persisted when the platelets were washed and resuspended in fresh medium and was also demonstrable in plasma. Platelet size was gradually increased by prostaglandin E1 (PGE1) which maintains platelets in a disc shape and does not cause the release of granule contents, indicating that the decrease in platelet density caused by PGE1 may be attributable to platelet swelling. PMID- 2982441 TI - Glycoprotein-180 deficiency: genetics and abnormal neutrophil activation. AB - Neutrophil function was studied in a patient with polymorphonuclear leukocyte (PMN) glycoprotein-180 deficiency and in her parents. PMNs of the patient had abnormal chemotaxis, phagocytosis, adherence, surface charge, and membrane associated events of activation. Selective defects to C3b, immunoglobulin G (IgG), phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl phenylalanine (FMLP) are described, although C3b receptor density was normal. The parents were found to have abnormal adherence to nylon-wool fibers, abnormal transmembrane potential depolarization with PMA, and reduced amounts of glycoprotein-180 in their PMNs. These studies provide further evidence that the oxidative burst has several different pathways for activation. They demonstrate that the absence of a single PMN surface glycoprotein is associated with a broad spectrum of PMN functional abnormalities. Finally, the observations made in the parents support an autosomal recessive mode of inheritance. PMID- 2982442 TI - Regulatory factors specific for adult and embryonic globin genes may govern their expression in erythroleukemia cells. AB - In order to test if trans-acting regulatory factors specific for globin genes of the adult and embryonic stages of development exist in erythroid cells, transcriptionally active embryonic and adult globin genes on the same chromosome were transferred by cell fusion from the human leukemia cell K562 into phenotypically adult mouse erythroleukemia cells. Restriction-fragment-length polymorphisms of the K562 zeta (embryonic) globin genes were used to establish that all three copies of human chromosome 16 present in the K562 cell showed the same pattern of human globin gene expression after transfer to the mouse erythroleukemia cell. Adult (alpha) but not embryonic (zeta) human globin mRNA was detected in all nine of the independently derived mouse erythroleukemia hybrid cells, each of which contained human chromosome 16. Restriction endonuclease studies of the K562 alpha- and zeta-globin genes after transfer into the mouse erythroleukemia cell showed no evidence of rearrangements or deletions that could explain this loss of zeta-globin gene expression. These data suggest that regulation of globin gene expression in these erythroleukemia cells involves trans-acting regulatory factors specific for the adult and embryonic stages of development. PMID- 2982443 TI - Pathology of peptide-producing neuroendocrine tumours. AB - The classic view of endocrinology as the study of a group of glands containing specialized endocrine cells, releasing substances that act at a distance, has been seriously questioned in recent years. The main reason for this was the discovery that endocrine cells are diffusely distributed through most body tissues and intermingled between nonendocrine elements. PMID- 2982444 TI - Malignant fibrous histiocytoma of the bladder. PMID- 2982445 TI - Granular cell myoblastoma of the bladder. PMID- 2982446 TI - Opportunistic viral infections. PMID- 2982447 TI - Recent progress in antiviral vaccines. PMID- 2982448 TI - Retroviral antigens and tumours. PMID- 2982449 TI - The Epstein-Barr virus as a model of virus-host interactions. PMID- 2982450 TI - The host response to herpes simplex virus. PMID- 2982451 TI - Chemotherapy for small cell lung cancer. PMID- 2982452 TI - Infection with parvovirus during pregnancy. PMID- 2982453 TI - AIDS and the health professions. PMID- 2982454 TI - Rehabilitation of patients with cardiac conditions. PMID- 2982455 TI - Chemotherapy for small cell lung neoplasms. PMID- 2982456 TI - Prolonged hypercalcaemia after industrial exposure to vitamin D3. PMID- 2982457 TI - Angiotensin II receptor localization in the canine CNS. AB - Specific binding of [125I]angiotensin II [(125I]Ang II) to sections of dog brain was determined by in vitro receptor autoradiography. Highly discrete, dark images representing specific binding of [125I]Ang II were observed in areas corresponding to the nucleus of the solitary tract, dorsal motor nucleus of the vagus, area postrema, ventrolateral medulla, pineal, subfornical organ, nucleus medianus, septum, organum vasculosum of the lamina terminalis and the anterior pituitary. The specific binding was frequently present either as a narrow band or tiny spot within a small portion of the nuclei to which the binding corresponded. The location of these Ang II recognition sites in regions associated with regulation of autonomic and neuroendocrine function provides further evidence for a role of this peptide within the central nervous system. PMID- 2982458 TI - The influence of opiate agonists on day-night feeding rhythms in young and old mice. AB - Daily rhythms of feeding behavior and responses to ketocyclazocine, morphine and naloxone were measured in young (1-2 months) and old (24-30 months) male CF-1 mice. All of the mice consumed more food at night than in the day-time, though this nocturnal peak was markedly reduced in the old animals, who consumed more in the day. The young mice also displayed a significant nocturnal enhancement in ketocyclazocine- and morphine-stimulated feeding. This day-night rhythm in ingestive responses was absent in the old mice. In comparison to the young mice, the opiate-stimulated food consumptions of the old animals were reduced at all times. Additionally, the old animals failed to show any day-night variations in the suppressive effects of naloxone on deprivation-induced food intake that were displayed by the young animals. PMID- 2982459 TI - The mechanism of cerebrovascular vasoconstriction in response to locus coeruleus stimulation. AB - Stimulation of the locus coeruleus in 11 Macaca nemestrina monkeys resulted in a frequency-dependent increase in internal carotid resistance (vasoconstriction) and a frequency-dependent decrease in external carotid resistance (vasodilation). The internal carotid constrictor response is maximal at low frequency stimulation (5 s-1), and decreases as frequency of stimulation is increased or decreased. It is blocked by the non-specific alpha-adrenoceptor blocker phentolamine and the alpha-2 adrenoceptor blocker yohimbine, but is unaffected by the alpha-1 adrenoceptor blocker prazosin. It is concluded that the cerebral vasoconstrictor response to locus coeruleus stimulation is mediated by alpha-2 adrenoceptors. PMID- 2982460 TI - Microinjection of neurotensin into the ventral tegmental area produces hypothermia: evaluation of dopaminergic mediation. AB - Neurotensin-producing perikarya and fibers have been identified in the ventral tegmental area of the rat, and recent microinjection studies indicate that neurotensin may function in the ventral tegmental area to regulate body temperature. In this study, the hypothermic response produced by intraventral tegmental injection of neurotensin was shown to be dose-dependent, with a threshold dose between 0.25 and 0.75 micrograms. When fluphenazine, a dopamine receptor antagonist, was microinjected into various forebrain nuclei simultaneous with neurotensin infusion into the ventral tegmental area, it was found to block neurotensin hypothermia. In contrast, injection with fluphenazine into the nucleus accumbens, lateral septum or preoptic area did not alter the hypothermic response. Furthermore, injection with atropine, phentolamine or diphenhydramine into the diagonal band of Broca did not block neurotensin hypothermia. Neurotensin was also injected directly into the preoptic region and shown to produce hypothermia. However, concurrent infusion of fluphenazine with neurotensin into the preoptic region did not attenuate neurotensin hypothermia. These data are consistent with the postulate that neurotensin acts in the ventral tegmental area to enhance dopamine release in the diagonal band of Broca, thereby producing hypothermia. However, neurotensin-induced hypothermia occurring after injection into the preoptic area does not appear to involve dopamine systems. PMID- 2982461 TI - Axon terminals of GABAergic chandelier cells are lost at epileptic foci. AB - Axon terminals of chandelier cells were analyzed in monkeys with cortical focal epilepsy produced by alumina gel to determine if this type of GABAergic terminal is lost at epileptic foci. These terminals form a dense plexus with the axon initial segments of pyramidal neurons, especially those in layers II and III. Axon initial segments of pyramidal neurons were traced for at least 40 micron in serial thin sections and beyond this point were observed to become myelinated. In single sections, 10-15 axon terminals were found to form symmetric synapses throughout the entire length of the axon initial segments from nonepileptic preparations and were observed to synapse with only these structures and not adjacent dendrites or spines. In epileptic cortex, the axon initial segments of pyramidal neurons were apposed by glial profiles that contained clusters of filaments typical of reactive astrocytes. Only a few, small axon terminals were observed to form symmetric synapses with these axon initial segments. Thus, the chandelier cell axons appeared to degenerate in epileptic cortex. The highly strategic site of GABAergic inhibitory synapses on axon initial segments suggests that they exert a strong influence on the output of pyramidal cells. The near absence of these chandelier cell axons in epileptic foci most likely contributes to the hyperexcitability of neurons. PMID- 2982462 TI - The influence of dopamine on epileptiform burst activity in hippocampal pyramidal neurons. AB - Dopamine (DA) application to guinea pig hippocampal CA1 neurons in vitro causes hyperpolarization of the resting potential, increase in conductance, and increase in amplitude and duration of the afterhyperpolarization (AHP). Since these changes could influence repetitive firing, we performed experiments to determine whether DA-induced effects would suppress epileptogenesis in the hippocampus. Epileptiform bursts were induced by adding penicillin (3.4 mM) to the perfusion medium. Focal application of DA (40-160 microns) onto CA1 cells (n = 15) produced a hyperpolarization averaging 4.5 mV beginning in 5-20 s and lasting up to 3 min. DA also caused an increase in the amplitude and duration of slow AHPs. The frequency of spontaneous epileptiform events however was not affected. CA3 neurons (n = 6) responded to DA application with an initial 1-3 mV depolarization beginning within 5-30 s and lasting 1-2 min. In 3 cases a small hyperpolarization lasting several minutes subsequently developed. AHP duration increased 70% and amplitude increased 35% (n = 4). Along with these membrane changes the frequency of epileptiform bursting in CA3 cells slowed for 1-3 min. We added DA (10-80 microM) to the perfusion medium to see whether a significant decrease in epileptiform burst frequency might occur in the follower CA1 region if greater numbers of pacemaker CA2 and CA3 cells were exposed to DA. Spontaneous CA1 bursting was reversibly slowed, the interburst interval became variable and increased from a mean of 4 to a mean of 5-7 s (n = 6). These results suggest that DA may play a role in decreasing the incidence or frequency of epileptogenic discharges in vivo. PMID- 2982463 TI - Studies on the in vivo release of vasoactive intestinal polypeptide (VIP) from the cerebral cortex: effects of cortical, brainstem and somatic stimuli. AB - The release of vasoactive intestinal polypeptide (VIP) from the surface of the sensorimotor (parietal) cortex of anesthetized cats was measured by radioimmunoassay (RIA) procedures. Two types of anesthetics were examined (chloralose-urethane and halothane). Increasing the dose or concentration of anesthetics produced a suppression of electrocortical activity and resulted in a reduction in the resting release of VIP from cerebral cortex. The mean rate of resting release of endogenous VIP in cats anesthetized with chloralose-urethane was 16.8 +/- 4.6 fmol/30 min/cm2 cortex. Although there were differences in the basal levels of VIP-like immunoreactivity (VIP-LI) among different animals, the baseline levels of VIP-LI varied little during the initial period of 6-7 h period of anesthesia with chloralose-urethane (40 and 300 mg/kg). Examination of representative electrocorticography which corresponded with the superfusion period revealed an association between stable levels and a relatively invariant pattern of electrical activity. All experimental manipulations were carried out during the period of stable release. The rate of release was increased by focal, unilateral stimulation of the cerebral cortex both ipsilateral and contralateral to the side of stimulation (3.2 +/- 1.1 and 1.4 +/- 0.5 times the resting release, respectively). Bilateral stimulation of the mesencephalic reticular formation (MRF) produced a frequency-dependent increase in cortical VIP release over the range of 60-100 Hz. Bilateral electrical stimulation of sciatic nerves at intensities that recruit A delta and C fibers failed to consistently increase the release of VIP-LI from the cortex in chloralose-urethane anesthetized cats but did evoke a significant increase of cortical VIP output in cats anesthetized with 1.5 and 1% halothane. Removal of calcium ions from the superfusing fluid and substitution of cobalt (a calcium channel blocker) or EDTA (a calcium chelating agent) for calcium ions did not affect the resting release of endogenous VIP, but attenuated the increase in VIP release normally evoked by electrical stimulation of the cortical surface or MRF. Tetrodotoxin (TTX), which blocks sodium channels and thus blocks the propagation of nerve impulses, did not influence the resting release of VIP. TTX, at concentrations that prevent the increase in VIP release evoked by veratridine, greatly enhanced the surface-stimulated release but abolished the increase in VIP release evoked by MRF stimulation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982464 TI - Influence of lateral septum and amygdala stimulation on the excitability of hypothalamic supraoptic neurons. An electrophysiological study in the rat. AB - Extracellular recordings were obtained from 116 phasically-active (putative vasopressinergic) and 113 continuously-active (putative oxytocinergic) neurosecretory neurons in the hypothalamic supraoptic nucleus of urethane or pentobarbital anesthetized male Sprague-Dawley rats. Single 1 Hz pulse stimulation in most regions of the amygdala and the ipsilateral lateral septum was followed by a transient (20-140 ms) reduction in the excitability of more than 90% of responsive cells; one third displayed a reduction in excitability to both amygdala and lateral septum stimulation. Amygdala or lateral septum stimuli delivered in brief trains of 20-100 pulses at 5-20 Hz during ongoing phasic discharges could induce silent periods lasting up to 30 or more seconds beyond the time of application. The same stimuli also reduced ongoing activity among continuously-firing SON cells but their response lasted only as long as the duration of the applied stimulus. These data suggest that neurons in both the ipsilateral septum and the various amygdaloid nuclei exert a predominantly inhibitory influence on the excitability of both vasopressinergic and oxytocinergic SON neurons in the rat. PMID- 2982465 TI - Epinephrine release and presynaptic alpha 2-adrenoceptic regulation in the guinea pig hypothalamus. AB - The release of epinephrine (E) and its regulation system were investigated in the slices of guinea pig hypothalamus. Electrical stimulation produced an efflux of [3H]E from tissues preloaded with [3H]E, in a current- and frequency-dependent manner. Stimulated [3H]E release was inhibited by tetrodotoxin and by a calcium free medium containing ethylene glycol bis(beta-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA). Stimulated [3H]E release was enhanced by yohimbine and this effect was suppressed by clonidine. These results provide strong evidence for the neurotransmitter role of E in the hypothalamus, and suggest the possible existence of regulatory mechanism of E release via presynaptic alpha 2-receptors. PMID- 2982466 TI - Spinal GABA receptors and central cardiovascular control. AB - Spinal superfusion with the GABA agonist muscimol reduced arterial pressure and heart-rate in the rat. Subsequent spinal administration of the GABA antagonist bicuculline restored blood pressure and heart-rate to control levels. Bicuculline alone transiently elevated blood pressure and produced a longer lasting tachycardia. Intrathecal injection of muscimol reduced sympathetic nerve discharge along with arterial pressure and cardiac rate. These results provide evidence for spinal GABA receptors participating in central cardiovascular regulation. PMID- 2982467 TI - Topographical organization of the dopamine-dependent adenylate cyclase of the chick embryo retina. AB - Retinas obtained from 9- and 16-day-old chick embryos and 5-day-old post-hatched chicken were analyzed with regard to the topographical distribution of the dopamine-stimulated adenylate cyclase system. The retinas were sectioned into 4 quadrants, namely ventroanterior, dorsoanterior, dorsoposterior and ventroposterior, taking the beak and the choroid fissure as references. Dopamine (0.1 mM) elicited the accumulation of cAMP in all 4 portions of tissue studied. However, when the ratio between the dopamine-stimulated vs the basal level of cAMP of 9-day-old embryo retina were compared in the different tissue portions, we observed that in the dorsoposterior quadrant the ratio was 15.5 while in the ventroanterior quadrant the ratio was approximately 6. The other two portions showed intermediate values. The same pattern was observed in retinas from 16-day old embryos. However, the ratio between the dopamine-stimulated vs the basal cAMP levels were 4.6 and 3 in the dorsoposterior and ventroanterior quadrants respectively. Further, the retina responsiveness to dopamine from post-hatched chicken was evenly distributed in the tissue, showing a stimulated/basal cAMP ratio of approximately 2 in all 4 quadrants studied. Our results show that the dopamine-dependent cAMP accumulation of the chick retina is unevenly distributed in the embryonic tissue and tends to homogeneity as the tissue differentiates. PMID- 2982468 TI - Effect of age on the rate of recovery of beta-adrenergic receptors in rat brain following desmethylimipramine-induced subsensitivity. AB - Brain tissues from aged rats have an impaired ability to increase beta-adrenergic receptors in response to reduced noradrenergic input, but can down-regulate these receptors in response to repeated administration of desmethylimipramine (DMI). In this study we compared the ability of brain tissues from young (3-month) and aged (20- to 26-month) rats to restore their density of beta-adrenergic receptors following desmethylimipramine (DMI)-induced receptor subsensitivity. Either DMI or saline was administered i.p. twice daily for 7 days to groups of young and aged rats. At various times after drug administration [3H]dihydroalprenolol (DHA) binding was determined in homogenates of pineal gland and cerebral cortex. Four hours after the last dose of DMI there was a decrease in DHA binding in both brain areas of young and aged rats. In young rats DHA binding in these tissues returned to control levels by 2 days after DMI administration. In contrast, in aged rats it took 8 and 16 days for DHA binding to recover in cerebrum and pineal, respectively. The concentration and half-life for the disappearance of DMI from serum and cerebrum were significantly greater in aged rats than in young rats, but the differences do not entirely explain the delayed recovery of beta receptors in the aged rats. The results suggest that beta-adrenergic receptors of brain tissues from aged rats cannot recover from beta-receptor subsensitivity as readily as those from young rats. If this recovery process requires the synthesis of new receptors, then this synthetic mechanism may be impaired with age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982469 TI - [PolyADP-ribosyl polymerase (adenosine diphosphate ribosyl transferase) in bovine crystalline lens; modulation of its activity during aging]. AB - PolyADP-ribose polymerase activities were measured in bovine lens. Activities similar to those in brain were found in the epithelial cells; none activity was detected in the fiber cells. During aging ADP-ribosyl transferase activity of epithelial cells raised, the number of polyADP-ribose chains increased while the average chain length decreased. A possible correlation between ADP-ribosylation and cell proliferation or repair is discussed. PMID- 2982470 TI - [Neonatal development of ACTH-immunoreactive neurons in the rat]. AB - Using immunohistochemical techniques, the distribution of ACTH 25-39 immunoreactive perikarya and fibers was studied in the newborn rat. Perikarya and fibers were demonstrated on day 1 after birth in the diencephalon, mesencephalon, pons and medulla. However, the amount of material increased rapidly from the first to third day of postnatal development. PMID- 2982471 TI - Accelerated onset of pancuronium with divided doses. AB - To determine the consequences of administering neuromuscular relaxants in divided doses, pancuronium was given either in a single dose, 0.07 mg X kg-1, or in an initial dose of 0.007 mg X kg-1 followed three minutes later with 0.063 mg X kg 1. When the drug was administered in divided dosage the onset time was reduced, the block was more intense and its duration of action was prolonged. It is suggested that such changes may be advantageous in the provision of rapid intense paralysis. PMID- 2982472 TI - Membrane engineering to rejuvenate the ageing brain. PMID- 2982473 TI - Endorphins and the neonate. PMID- 2982474 TI - Combined surgery, intraoperative brachytherapy, and postoperative external radiation in stage III non-small cell lung cancer. AB - From March 1977 to December 1980, 318 patients with Stage III non-small cell lung cancer underwent thoracotomy at Memorial Sloan-Kettering Cancer Center. One hundred of these patients, considered for this study, were treated by a multimodality approach consisting of resection and/or intraoperative brachytherapy followed by postoperative external irradiation. The criteria for utilizing intraoperative brachytherapy and postoperative external irradiation were either the presence of residual gross disease (47%) or close resection margins suspected to be involved by the tumor (53%). The intraoperative brachytherapy consisted of a temporary iridium 192 implant for subclinical disease in the mediastinum (median dose, 30 Gy in 3-5 days) and a permanent iodine 125 implant for residual gross disease usually at the primary site (median dose, 160 Gy). All patients received postoperative external beam irradiation consisting of 30 to 40 Gy in 2 to 4 weeks. Seven patients (7%) experienced mild to severe complications after these treatments. The local control, when all gross disease had been removed, was influenced by the presence or absence of tumor at the margins of resection (53% and 89%, respectively). The local control in the patients with gross residual disease treated by brachytherapy and postoperative external irradiation, (40 Gy in 4 weeks) was 72%. The actuarial overall 5-year survival was 22%. The 5-year survival was better in patients who had all gross disease removed as compared with patients who had gross residual disease (30% versus 13%). The disease-free survival in these two groups was 27% and 12%, respectively. This review shows that complete resection and moderate doses of postoperative external irradiation achieves a satisfactory local control and improves survival. When all gross disease cannot be removed, then brachytherapy and postoperative external irradiation may achieve similar local control. Distant metastases still remain a major problem in these patients. PMID- 2982475 TI - Biologic characteristics of four Ewing's sarcomas. AB - Four Ewing's sarcomas were examined for chromosomal characteristics, growth in cell culture, tumorigenicity in nude mice, and presence of beta-adrenergic receptors. Three tumors were from untreated patients (one obtained directly from the patient and two after growth in nude mice) and one was a metastatic lesion obtained after treatment. All four tumors were diploid or near-diploid, with one or more structural rearrangements. In the metastatic lesion, 21 abnormalities were seen. No specific chromosome aberration was found to be common to all four tumors, although a t(1,16) was observed in two and a t(11;22) in two. Abnormalities involving chromosomes #1, #3, #11, #13, #16, and #22 were each found in two of four tumors. All four tumors were tumorigenic in nude mice; two grew well in cell culture, one of which became an established line, and all four expressed high concentrations of beta-adrenergic receptors. PMID- 2982477 TI - Cytogenetic analysis of hepatoblastoma. AB - A cytogenetic investigation was performed in a case of hepatoblastoma; the analyses revealed a pseudodiploid karyotype. PMID- 2982476 TI - Karyotypic characterization of established cell lines and short-term cultures of human lung cancers. AB - Karyotypic patterns were analyzed from the four major histopathologic groups of human lung cancer: small cell (SCC), squamous cell (SQC), large cell (LCC), and adenocarcinoma (ADC). The studies were performed on banded chromosomes from direct preparations of pleural fluids (one case of SQC and LCC, respectively) and on cell lines. All metaphases were aneuploid and showed highly rearranged chromosomes, with the exception of the direct preparation of the SQC, which was pseudodiploid. The number of marker chromosomes varied-from tumor to tumor. No consistent aberrations could be detected. Special attention was paid to chromosomes 3p-, which was earlier reported to be a characteristic marker chromosome for SCC. We could confirm the presence of that abnormality in two of our six SCC lines. However, we also found a 3p- in a primary SQC culture, in one LCC cell line, and in one ADC cell line. The breakpoint on 3p was not consistent. In some lines, numerical and structural changes of chromosomes #1, #12, #14, and #22 were also noteworthy, although none of these chromosome abnormalities seemed to be correlated to a certain histopathologic group. PMID- 2982478 TI - A 3p deletion in small cell lung carcinoma. AB - A specific chromosomal abnormality (3p del) was found in direct preparations from three small cell carcinomas of the lung: one primary tumor and two metastatic tumors in mediastinal lymph nodes. This lends support to Whang-Peng et al.'s [3] detection of the deletion in tissue cultures. PMID- 2982479 TI - Hydrogen peroxide and hydroxyl free radical production by hematoporphyrin derivative, ascorbate and light. AB - Photofrin II, a commercial preparation of a hematoporphyrin derivative containing a high proportion of the active di-hematoporphyrin ether, was found to convert oxygen to hydrogen peroxide and hydroxyl free radicals in the presence of ascorbate and light. In addition, a light-dependent increase in the ascorbate free radical concentration was observed in hematoporphyrin derivative/ascorbate solutions with subsequent oxidation of ascorbate. PMID- 2982480 TI - Potentiation of cytotoxic activity of immunotoxins on cultured human cells. AB - The cytotoxic activity against human tumor cells of toxic conjugates of Pseudomonas exotoxin with anti-transferrin receptor antibody or epidermal growth factor was potentiated up to 10 to 20-fold by the calcium antagonists verapamil, D-600, and diltiazem and by the lysosomotropic agent beta-glycylphenyl naphthylamide. The potentiating activity of these agents could be predicted by measuring the inhibition of protein synthesis by the immunotoxins on various cell lines. The use of potentiating agents such as these in combination with immunotoxins may prove useful in the treatment of some human cancers. PMID- 2982481 TI - Comparison of the actions of RU 38486 and megestrol acetate in the model of a transplantable adrenocorticotropin- and prolactin-secreting rat pituitary tumor. AB - The effects of the progesterone antagonist RU 38486 and the progesterone agonist megestrol acetate on the growth of the estrogen-progesterone receptor-positive transplantable adrenocorticotropin (ACTH)/prolactin-secreting rat pituitary tumor 7315a were examined. RU 38486 (2.5 mg/kg/day) for 30 days significantly inhibited tumor size, tumor weight, and the plasma prolactin and ACTH concentrations, while the same dose of megestrol acetate only inhibited pituitary tumor weight. Megestrol acetate inhibited both the release and total ACTH content of the anterior pituitary gland, while RU 38486 increased both the release and the total ACTH content. Studies with ACTH secretion by cultured normal rat pituitary cells showed that megestrol acetate (1 microM) did not affect corticotropin-releasing factor (CRF)-stimulated ACTH release after 4-hr exposure, but inhibited CRF stimulated ACTH release by 50% after 24-hr preincubation. The glucocorticoid-like effect of 1 microM megestrol acetate in this model is similar to that exerted by 10 nM dexamethasone. Acute exposure or preincubation of rat pituitary cells with RU 38486 (1 microM) did not influence CRF-stimulated ACTH release, while preincubation for 24 hr revealed a dose-dependent reversing effect of RU 38486 on dexamethasone-induced inhibition of CRF-stimulated ACTH release. In this model, 1 microM RU 38486 completely overcame the effect of 10 nM dexamethasone. Megestrol acetate and RU 38486 have inhibitory effects on the growth of the 7315a tumor. They differ both with regard to their effects on the progesterone and the glucocorticoid receptor, with megestrol acetate exerting an agonistic and RU 38486 an antagonistic action. PMID- 2982482 TI - Selective cytotoxicity of the SM1 monoclonal antibody towards small cell carcinoma of the lung. AB - A murine monoclonal antibody, SM1, is strongly reactive with the surface membrane of small cell carcinoma of the lung. SM1 antibody is unreactive with most other cancers and various normal tissues including bone marrow cells. We now find that SM1 antibody is selectively cytotoxic to small cell carcinoma (SCC) in vitro. The antibody is present in high titers in supernatant fluids or ascites obtained by i.p. injection of SM1 hybridoma cells into pristaned BALB/c mice. The cytotoxic effect of the antibody is reduced to one-half maximal activity only at dilutions greater than 1:40,000. The efficiency of tumor cell lysis is greatly enhanced by repeated treatments with antibody and complement. Using three treatments with antibody and complement, 99.9% of SCC cells are lysed, as determined by the chromium release. Similar efficiency of SCC cell kill was observed by clonogenic assays. SM1 antibody produces no significant antibody-dependent lysis of cell lines derived from non-SCC lung carcinomas and leukemia cells. The results from chromium release assay and clonogenic assays also indicate that the effect of SM1 antibody and complement on bone marrow cells is minimal and could be accounted for by the effect of complement alone. PMID- 2982483 TI - Effect of difluoromethylornithine, an inhibitor of polyamine biosynthesis, on the topoisomerase II-mediated DNA scission produced by 4'-(9 acridinylamino)methanesulfon-m-anisidide in L1210 murine leukemia cells. AB - Treatment of mouse leukemia L1210 cells with the polyamine biosynthesis inhibitor alpha-difluoromethylornithine (DFMO) increased the magnitude of the DNA scission produced by the DNA intercalator 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA). This enhanced DNA scission was protein concealed and protein associated, as was the m-AMSA-induced scission in cells unexposed to DFMO. The effect of DFMO required more than 6 hr to develop and was greater at 48 hr than at 24 hr of exposure to DFMO. Exogenously added putrescine partially reversed the effects of DFMO, while exerting no effect on m-AMSA-induced DNA scission in cells unexposed to DFMO. The cellular uptake of [14C]-m-AMSA was the same in DFMO treated or untreated cells. The DNA scission and DNA-protein cross-linking produced by m-AMSA appear to represent the stabilization of an intermediate in the normal cycle of topoisomerase II function (Nelson, E.M., Tewey, K.M., and Liu, L.F., Proc. Natl. Acad. Sci. USA, 81: 1361-1365, 1984). Since polyamine depletion appears to affect the magnitude of this effect in cells, and since polyamines can alter topoisomerase II function in vitro, polyamines may be involved in topoisomerase function in vivo either directly or through secondary effects, such as alterations of the conformation of chromatin, the intracellular site at which topoisomerase acts. PMID- 2982484 TI - Tumor-associated antigens on bovine leukemia virus-induced bovine lymphosarcoma identified by monoclonal antibodies. AB - Thirteen monoclonal antibodies directed against tumor cells from cattle from enzootic bovine leukosis (EBL) were obtained. They reacted with tumor cells but not with normal bovine cells or bovine leukemia virus antigens. According to the reactivities of these antibodies with 19 individual tumors, the 13 monoclonal antibodies can be divided into three groups. Antibodies of the first group reacted with all the EBLs tested; those of the second group reacted with several, but not all, of the EBLs tested; and those of the third group reacted only with homologous tumor cells. Therefore, tumor-associated antigen (TAA) on the EBL tumor may possess common TAA, partially common TAA, and individually distinct TAA. The TAAs were solubilized from the tumor cells by treatment with 0.2% sodium deoxycholate and partially purified by diethylaminoethyl-cellulose column chromatography. Eleven of the 13 monoclonal antibodies reacted with this soluble TAA. The monoclonal antibodies belonging to the first group inhibited in vitro the growth of the bovine lymphoid cell line derived from the EBL tumor. PMID- 2982485 TI - Collagen synthesis by short-term explants of pediatric tumors. AB - Collagens are a heterogeneous family of structural proteins synthesized by many cultured cells including tumor cells. The synthesis of these proteins by three human tumor types commonly encountered in children [neuroblastoma, rhabdomyosarcoma, and nephroblastoma (Wilms' tumor)] was investigated in short term cultures of freshly excised tumor explants grown on extracellular matrices. Analysis of the incorporation of [3H]proline into collagenase-sensitive proteins indicated significant collagen production by several Wilms' tumors and rhabdomyosarcomas, while neuroblastomas did not synthesize this structural protein. All eight Wilms' tumor specimens analyzed secreted type IV procollagen. Interstitial types I and III collagens were also produced by these tumors, but in most cases, the alpha 1 (I): alpha 2 ratio was much higher than the 2:1 ratio expected for type I collagen, indicating a major change in the control of type I collagen production. Rhabdomyosarcomas were very heterogeneous with regard to collagen secretion and synthesized either a single collagen isotype (type III), several collagens including types I, III, and IV, or no detectable collagen. Our data represent a first quantitative and qualitative analysis of collagen synthesis by primary tumor cultures and reveal much more heterogeneity in collagen biosynthesis by these tumors than reported previously with established cell lines. They also indicate significant alterations in the expression of type I collagen genes in Wilms' tumors. PMID- 2982486 TI - Establishment, characterization, chemosensitivity, and radiosensitivity of two different cell lines derived from a human breast cancer biopsy. AB - In vitro culture of a human breast cancer biopsy fragment gave rise to two permanent cell lines, CAL 18 A and CAL 18 B, which were differentiated by both morphological and ultrastructural analysis. The karyotypic and growth properties of these two cell lines also differed, providing further evidence of cell heterogeneity within a given tumor. Both cell lines lost their hormone receptors in vitro. CAL 18 A cells grew in agar and were tumorigenic after inoculation into nude mice; neither of these properties was observed in CAL 18 B cells. The chemosensitivity of 12 antineoplastic drugs was assessed by a short-term assay, using inhibition of tritiated thymidine incorporation by the cells after contact with the drugs as the end point. Only a few drugs were active at moderate concentrations. The overall responses of both cell lines were similar. The cell survival curves, established by the colony method following a single dose of radiation, were also very similar, despite the greater heterogeneity of CAL 18 B cells. The two cell lines appear to be interrelated, since CAL 18 B cells were occasionally observed to emerge from CAL 18 A clones, suggesting that malignant cell redifferentiation may occur spontaneously in vitro. PMID- 2982487 TI - Ductal metaplasia of human exocrine pancreas and its association with carcinoma. AB - Monoclonal antibodies to cell surface markers of human exocrine pancreas were used to establish the cytotypic expression of cells forming "tubular complexes" in pancreases from six adults without carcinoma and in the nontumorous pancreatic parenchyma of 16 pancreases with carcinoma. These cells manifested duct cell determinants. In general, the presence of cells with duct cell surface markers within the acini corresponded to the normal distribution of centroacinar cells in the 30 control human pancreases (from cadaveric donors); however, foci of abnormal acini were seen in these pancreases independent of or intermingled with the "tubular complexes." The acini in these abnormal areas were formed by a core of cells and cell processes that expressed duct cell determinants. They were partially surrounded by acinar cells and showed slight or no lumenal dilation. While the causative agent(s), the cell(s) of origin, and the regression and/or progression of these lesions are yet to be determined, the replacement of acini by the spectrum of lesions composed of cells with duct cell surface marker is suggested to constitute ductal metaplasia. PMID- 2982488 TI - Effect of cholecystokinin on human cholangiocarcinoma xenografted into nude mice. AB - Gastrointestinal polypeptide hormones regulate growth of various normal gastrointestinal tissues as well as certain visceral cancers. Since cholecystokinin (CCK) promotes growth of normal biliary tract, we sought to determine whether CCK affects the growth and metabolism of human cholangiocarcinoma line SLU 132. Twenty-six nude mice with s.c. xenografts of this cancer received either CCK octapeptide (50 micrograms/kg/dose) or 0.9% NaCl solution (saline) twice a day i.p. for 14 days. Tumor volume was calculated from Vernier caliper measurements. At sacrifice on Day 15, tumors were excised, weighed, and examined histologically. DNA, RNA, and protein were measured in the xenografted carcinomas. Because this cholangiocarcinoma produces carcinoembryonic antigen (CEA), we obtained serum at sacrifice for CEA radioimmunoassay and also tumor tissue for CEA immunolabeling with murine anti-CEA monoclonal antibody. Serum CEA levels were 90% higher in the CCK-treated group. Tumor tissue in the CCK-treated group also contained more CEA than did the controls. Mean tumor volume increased significantly in the saline group during the 14-day treatment period, whereas mean tumor volume did not increase significantly in the CCK group. Exogenous high-dose CCK thus appears to increase production and release of CEA from SLU-132; it also appears to retard growth of this tumor line in the nude mouse. PMID- 2982489 TI - Purification and characterization of high-affinity cyclic adenosine 5' monophosphate phosphodiesterases from human acute myelogenous leukemic cells. AB - Soluble, high-affinity cyclic adenosine 3':5'-monophosphate (cyclic AMP) phosphodiesterases extracted from blast cells of patients with acute myelogenous leukemia have been characterized by physical, kinetic, and immunological criteria and fractionated to a high degree of purity. Procedures used in this study were similar to those used to purify the high-affinity enzyme from dog kidney. Two forms of high-affinity enzyme were found in blast cells. Form A was similar to the known type IV phosphodiesterases, including those of normal lymphocytes and monocytes. It showed a molecular weight near 60,000, a rate of hydrolysis of cyclic AMP 7 to 10 times that of cyclic guanosine 3':5'-monophosphate (cyclic GMP), competitive inhibition by cyclic GMP for cyclic AMP hydrolysis, and identical immunoreactivity by Western transfer analysis. This enzyme form was purified to apparent homogeneity by physical criteria but showed a low maximum velocity relative to other phosphodiesterase forms. A second, different form of high-affinity phosphodiesterase (Form B) was also resolved and partially purified. By comparison with Form A, this enzyme eluted from diethylaminoethyl cellulose at slightly lower ionic strength, had a lower molecular weight, appeared specific for cyclic AMP as substrate, showed no inhibition of cyclic AMP hydrolysis by cyclic GMP, and displayed no immunological cross-reactivity to the Mr 60,000 enzyme. Neither enzyme form was activated by calmodulin or proteolysis, whereas both showed comparable inhibition by 6,7-dimethoxy-1 veratrylisoquinoline, 1-methyl-3-isobutylxanthine, and 1,3-dimethylxanthine. PMID- 2982490 TI - Intensive alternating chemotherapy regimen in small cell carcinoma of the lung. AB - Fifty-five patients with small cell carcinoma of the lung (SCCL) were treated with alternating courses of CAV (cyclophosphamide, doxorubicin, and vincristine) and VHM (etoposide, hexamethylmelamine, and methotrexate) at 21-day intervals. One regimen contained high-dose cyclophosphamide (2400 mg/m2 iv), while the second contained an increased dose of etoposide (250 mg/m2 iv on Days 1-3). Patients achieving a complete response (CR) received a minimum of six cycles of therapy or two cycles beyond the achievement of CR. These patients as well as those with partial response with disease limited to the lung then received radiotherapy to the lung primary (5100 rads in 300-rad fractions) as well as prophylactic cranial radiotherapy (3000 rads in 300-rad fractions). All therapy was subsequently stopped until relapse. There were no deaths during the aplastic periods induced by the intensive chemotherapy. CR was achieved in 57% of the patients and partial response was achieved in 38%. The actuarial median survival for the 27 patients with extensive disease was 13.5 months and for the 28 patients with limited disease was 13 months. Survival at this point appears comparable to less intensive regimens in which maintenance therapy was given. We conclude that prolonged intensive induction chemotherapy in SCCL is well tolerated. Studies are needed to examine in a randomized trial the role of alternating combinations and maintenance therapy in SCCL. PMID- 2982491 TI - Effect of adjuvant central iv hyperalimentation on the survival and response to treatment of patients with small cell lung cancer: a randomized trial. AB - The effect of central iv hyperalimentation (IVH) as an adjunct to aggressive antineoplastic therapy for small cell carcinoma of the lung was evaluated in a randomized trial with 119 evaluable patients. IVH was given over a 28-day period with higher caloric and protein intake for patients nutritionally depleted on entry in the study; all patients were escalated in caloric and protein intake to maximize nutritional repletion. Combination chemotherapy and radiation therapy induced a 45.5% complete response rate and an overall response rate of 92.8%. Median survival for patients with limited disease was 18 months; median survival for patients with extensive disease was 11 months. Patients randomized to receive IVH did not have a better response rate (P = 0.97) or survival (P = 0.78) than control patients. IVH did not significantly alter the survival for patients who at baseline had greater than 5% pretreatment weight loss, low caloric intake, decreased serum albumin, or reduced total iron-binding capacity. Significantly more febrile episodes were seen in IVH patients than in control patients (P less than 0.001). Short-term IVH to patients with this malignancy who are capable of enteral alimentation cannot be routinely recommended as adjunctive therapy. PMID- 2982492 TI - Probable cure of small cell carcinoma of the lung by etoposide. PMID- 2982493 TI - Cell-cycle-specific activity of cyclic nucleotide phosphodiesterase in Physarum polycephalum. AB - The cell-cycle-related activities of the cAMP- and cGMP-dependent phosphodiesterases of Physarum polycephalum were assayed. The activities of plasmodial homogenate and of selected subcellular fractions were measured. The results suggested the presence of both cAMP- and cGMP-dependent phosphodiesterase in the isolated nuclei of P. polycephalum. In addition, they reveal that the cAMP and cGMP-dependent phosphodiesterase activities of the subcellular fractions fluctuate throughout the cell cycle. The whole-cell homogenates exhibit no cell cycle-related changes in the presence of 5 X 10(-4) M cGMP. Kinetic data suggest the presence of multiple phosphodiesterase activities in the homogenate and its particulate fractions for the cGMP-dependent enzyme. Multiple cAMP activities are also suggested for the particulate fractions. The Km values indicate that the substrate affinities of the phosphodiesterases from P. polycephalum are similar to those found previously in mammalian systems. PMID- 2982494 TI - Apurinic sites as mutagenic intermediates. PMID- 2982495 TI - Ty elements transpose through an RNA intermediate. AB - We have followed Ty transposition with a donor Ty element, TyH3, whose expression is under the control of the GAL1 promoter. Sequence analysis reveals dramatic structural differences in TyH3 before and after transposition. If the donor TyH3 is marked with an intron-containing fragment, the intron is correctly spliced out of the Ty during transposition, suggesting that the Ty RNA is the intermediate for transposition. Furthermore, the pattern of sequence inheritance in progeny Ty insertions derived from the marked Ty follows the predictions of the model of retroviral reverse transcription. Comparison of marked Ty elements before and after movement shows that transposition is highly mutagenic to the Ty element. These results demonstrate that during transposition, Ty sequence information flows from DNA to RNA to DNA. PMID- 2982496 TI - Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency. AB - The thymidine kinase gene of herpes simplex virus 1 was mutated by inserting oligodeoxynucleotide linkers into the region of the gene corresponding to the 5' untranslated portion of the mRNA. These linkers, when transcribed into mRNA, might be expected to form hairpin loops and hence to increase the secondary structure of the 5' end of the mRNA. Thymidine kinase insertion derivatives were examined in vivo and in vitro to determine translational efficiency. For the in vivo studies, thymidine kinase insertion derivatives were transfected into thymidine kinase deficient L cells alone and together with a selectable dominant marker, or were assayed in the COS-1 transient expression system. For in vitro studies, thymidine kinase insertion derivatives were subcloned into pSP64. Capped transcripts were analyzed for their ability to bind ribosomes and translate in rabbit reticulocyte lysates and wheat-germ extracts. The results demonstrate that translation efficiency is decreased as the number of linkers is increased and support the view that excessive secondary structure at the 5' end of eukaryotic mRNA impedes translation. PMID- 2982497 TI - Rous sarcoma virus encodes a transcriptional activator. AB - Rous sarcoma virus expresses a transcriptional activator that affects the LTR as well as other promoters. We discern this activity as a stimulation of the transient expression of an LTR-promoted hybrid transcriptional unit and also of the rat preproinsulin II gene in transfected NIH 3T3 cells. We map the activity to an alternate reading frame in the p19-p10 region of the gag gene and identify a mRNA whose spliced structure would direct translation of this reading frame from the Pr76gag initiation codon. This mRNA probably differs from genomic RNA only by the 282 nucleotide splice. The predicted translation product is a 124 residue polypeptide; the first six amino acids arise from gag. The target for the action of this transcriptional modulator at the LTR lies between 111 and 620 nucleotides upstream of the cap site. PMID- 2982498 TI - Monoclonal antibodies NORM-1 and NORM-2 induce more normal behavior of tumor cells in vitro and reduce tumor growth in vivo. AB - In this study, large numbers of hybridomas (produced by syngeneic immunization with B16 mouse melanoma and fusion with NS-1 myeloma cells) were screened for the production of antibodies that affected morphology and growth of animal and human tumor cells in vitro. Two such antibodies, NORM-1 and NORM-2 (both IgG2a), inhibited the growth of B16 melanoma cells in soft agar and increased the serum requirements of tumor cells in tissue culture. Antibody NORM-2 also inhibited the growth of SV40-transformed 3T3 cells in agar and caused them to deposit more fibronectin into extracellular matrix. These antibodies thus seem to induce a more normal behavior of tumor cells in vitro. In vivo both antibodies reduced the number of growing lung tumors of B16 melanoma in C57BL/6 mice by 70%-90% when injected 3 days after the tumor cells. By immunoprecipitation of 35S-methionine labeled cell extracts, NORM-2 antibody recognized a 59 kd protein in B16 mouse and in A375 human melanoma cells but not in 3T3 fibroblasts. PMID- 2982499 TI - The Drosophila EGF receptor gene homolog: conservation of both hormone binding and kinase domains. AB - Chicken v-erB probe was used to isolate a unique clone of Drosophila melanogaster DNA. It maps by in situ hybridization to position 57F on chromosome 2. A complete nucleotide sequence of the coding region has been obtained. The putative Drosophila EGF receptor protein is similar in overall organization to the human homolog. It shows three distinct domains: an extracellular putative EGF binding domain, a hydrophobic transmembrane region, and a cytoplasmic kinase domain. The overall amino acid homology is 41% in the extracellular domain and 55% in the kinase domain. Two cysteine-rich regions, a hallmark of the human ligand-binding domain, have also been conserved. Fusion of the coding sequences of the kinase and extracellular domains generating the receptor gene must have occurred over 800 million years ago. PMID- 2982500 TI - Protein phosphorylation at tyrosine is induced by the v-erbB gene product in vivo and in vitro. AB - The v-erbB gene product of avian erythroblastosis virus (AEV) has extensive homology with the receptor for epidermal growth factor (EGF). We report here that chicken embryo fibroblasts (CEF) transformed by AEV show enhanced tyrosine phosphorylation of a number of cellular polypeptides, including the 36 kd protein, which is phosphorylated in avian sarcoma virus-transformed fibroblasts, and the 42 kd protein, which is phosphorylated in mitogen-stimulated cells. CEF infected by AEV mutants with deletions in v-erbA showed enhanced tyrosine phosphorylation, whereas CEF infected by mutants with deletions in v-erbB did not. When membranes from AEV-transformed cells were incubated with gamma-32P-ATP, both the v-erbB gene product and the 36 kd cellular protein became phosphorylated at tyrosine. These results indicate that the v-erbB protein induces tyrosine phosphorylation in vivo and in vitro, and suggest that, like the EGF receptor, it possesses tyrosine-specific protein kinase activity. PMID- 2982501 TI - Antibodies against a synthetic peptide as a probe for the kinase activity of the avian EGF receptor and v-erbB protein. AB - The transforming protein v-erbB of avian erythroblastosis virus (AEV) displays extensive sequence homology with the presumptive protein-tyrosine kinase domain of the human EGF receptor and with the src protein-tyrosine kinase family of oncogenes. However, no kinase activity has previously been demonstrated for the v erbB protein. Here antibodies generated against a synthetic peptide from the C terminus of human EGF receptor are shown to immunoprecipitate the EGF receptor from human and avian cells, as well as the v-erbB proteins from AEV-transformed cells that become phosphorylated on tyrosine residues upon the addition of gamma 32P-ATP. The immunoprecipitates are also able to phosphorylate exogenous tyrosine containing substrates. Hence, it is likely that both avian EGF receptor and v erbB proteins are protein tyrosine-specific protein kinases. Since the kinase activity of v-erbB protein cannot be regulated by EGF, it is proposed that the tyrosine protein kinase function of v-erbB may be constitutively activated. PMID- 2982502 TI - Monoclonal antibody to a membrane glycoprotein inhibits the acrosome reaction and associated Ca2+ and H+ fluxes of sea urchin sperm. AB - Monoclonal antibodies were prepared using sea urchin (Strongylocentrotus purpuratus) sperm or isolated sperm plasma membrane vesicles as immunogen. Two monoclonal antibodies, J4/4 and J10/14, react with different epitopes on external domains of the same Mr = 210,000 integral-membrane glycoprotein. At Fab fragment concentrations of equivalent cell surface binding, J10/14 inhibits the egg jelly induced exocytotic acrosome reaction and associated Ca2+ influx and H+ efflux, whereas J4/4 does not. Immunofluorescent localization shows that both monoclonal antibodies react with a narrow collar of plasma membrane over the acrosomal complex and also with the entire flagellum. PMID- 2982503 TI - Adenovirus E1a proteins repress transcription from the SV40 early promoter. AB - Using a transient expression assay in HeLa cells, we show that products from the adenovirus-5 E1a transcription unit repress transcription from the SV40 early promoter. The repression is unrelated to T antigen autoregulation, occurs maximally with low concentrations of E1a expression plasmid, is exerted at the transcriptional level, and requires functional E1a protein. The 289 and 243 amino acid E1a proteins are equally effective at repressing transcription. Since only the 289 amino acid protein is efficient at activating transcription, we conclude that activation and repression are separate E1a functions. We discuss possible mechanisms for E1a repression and the relationship of repression to the function of E1a in cell immortalization and transformation. PMID- 2982504 TI - The early (18-hr) human mixed lymphocyte reaction: identification and isolation of activated T-cell clones. AB - This study provides direct evidence that de novo expression of the activation antigens B1 49.9 (49.9) (interleukin-2 receptor) and 4F2 enables identification of alloactivated cells within 18 hr of initiation of human mixed lymphocyte reactions (MLR). Using a dual-parameter flow cytometer (simultaneous assessment of immunofluorescence and DNA content on the same cell), it was demonstrated that these activation antigens emerge before activated cells enter into S/G2/M phase of the cell cycle. Family studies illustrate that early activation antigen appearance occurs in response to a mismatch at chromosome 6, and invariably heralds the proliferative outcome at 6 days of MLR. In order to directly study the small alloactivated T-cell population, 49.9-positive cells were isolated using a cell sorter after 18 hr of MLR and cloned by limiting dilution using purified recombinant interleukin-2 (rIL-2). Antigen-specific T4-positive T-cell clones were isolated. Analysis of these clones demonstrates that antigen-specific reactivity is acquired within 18 hr in the MLR. These methods should permit a dissection of the early events of alloactivation. PMID- 2982506 TI - Restorative resin use in 1985--state of the art. PMID- 2982505 TI - In vivo effects of cyclosporine on influenza A virus-infected mice. AB - Cyclosporine (cyclosporin A, CsA) administered to mice substantially affects their immune response to an influenza A virus infection. If treated with CsA for 21 days, the mouse lungs contain high titers of virus which are cleared more slowly than in controls. Indicators of pathological damage--lung weight, extent of consolidation, fine morphology, and the extent of infiltration of dividing cells into the lung--showed that administration of CsA greatly decreased the level of inflammation. The production of hemagglutination-inhibiting (HI) antibody was delayed but reached almost control levels and NK cell activity in the lung was also comparable to control levels. In contrast, a delayed-type hypersensitivity (DTH) response to the virus could not be elicited in the CsA treated, infected mice at 6 or 12 days after infection. Cytotoxic-T-cell (Tc cell) activity was present in the lungs of such mice though its appearance was delayed and the activity recovered was less than that of the control infected mice. If administered with a dose of virus lethal for normal mice. CSA-treated mice survived, probably due to the greatly reduced level of immunopathological damage in the infected lung. PMID- 2982507 TI - Quantitative structure-activity relationships of cysteine hydrolases. Ficin hydrolysis of X-phenyl-N-methanesulfonyl glycinates. AB - The hydrolysis of ficin of 33 X-Phenyl-N-methanesulfonyl glycinates has been studied. The resulting Km-values have been used to derive a quantitative structure-activity relationship (QSAR). The QSAR for ficin is compared with QSAR for other cysteine hydrolases. The comparisons show that although there are specific differences, overall the reaction mechanisms are very similar. PMID- 2982508 TI - Effect of inhibitors of alpha-naphthylisothiocyanate-induced hepatotoxicity on the in vitro metabolism of alpha-naphthylisothiocyanate. AB - The role of S-oxidation in the toxic bioactivation of alpha naphthylisothiocyanate (ANIT) was investigated. The effects of several thione compounds, inhibitors and an inducer of the cytochrome P-450-dependent mixed function oxidase systems on the in vitro metabolism of ANIT and aminopyrine were determined. Ethionamide, sodium diethyldithiocarbamate (Na-DDTC) and S-methyl diethyldithiocarbamate (Me-DDTC), three agents known to undergo metabolism by an S-oxidative pathway and diminish ANIT's toxicity, inhibited the in vitro enzymatic metabolism of ANIT by rat liver microsomes. Methimazole failed to alter either the hyperbilirubinemic response of ANIT or the in vitro metabolism of ANIT. All four thione compounds (i.e., ethionamide, Me-DDTC, Na-DDTC and methimazole) inhibited the enzymatic metabolism of aminopyrine by rat liver microsomes. Me-DDTC was the most potent, whereas methimazole was the least potent inhibitor of aminopyrine metabolism. Phenobarbital, which potentiates, and SKF 525A, which inhibits the hepatotoxicity of ANIT in vivo, correspondingly stimulated or inhibited the NADPH-dependent metabolism of ANIT and aminopyrine by liver microsomes. N-Decylimidazole (NDI), another classical inhibitor of cytochrome P-450-dependent monooxygenase system, inhibited both the in vivo toxicity and in vitro metabolism of ANIT. NDI also diminished the enzymatic metabolism of aminopyrine by liver microsomes. Thus the results of this study indicate that metabolism of ANIT is intimately related to its toxicity and that ANIT probably undergoes its toxic bioactivation via a cytochrome P-450-dependent S-oxidative pathway. PMID- 2982509 TI - High-dose cyclophosphamide with autologous bone marrow rescue after conventional chemotherapy in the treatment of small cell lung carcinoma. AB - Within an original consecutive series of 94 patients, 36 eligible patients with small cell lung carcinoma were treated with high-dose cyclophosphamide 7 g/m2 after conventional chemotherapy with VP16, adriamycin, and vincristine. The first 17 also underwent autologous bone marrow rescue. Treatment was well tolerated apart from one treatment-related death. Measurable tumour was still present in 15 patients before high-dose cyclophosphamide, and although 12 (80%) of these achieved further tumour response, these responses were all short-lived, with a median duration of 9 weeks. In 14 limited-disease patients already in complete remission before high-dose therapy the initial result was better, but 11 (79%) have now relapsed following overall median response duration of 10 months. High dose cyclophosphamide after conventional chemotherapy is feasible and achieves a high response rate, but it does not appear to be associated with significant survival benefit either overall or in patient subgroup. PMID- 2982510 TI - Effect of nitrosoureas on calmodulin activity in vitro and in mouse intestine in vivo. AB - The effects of BCNU, CCNU, methyl-CCNU, streptozotocin, and chlorozotocin on calmodulin activity were studied in vitro and in vivo. Preincubation of BCNU, CCNU, and methyl-CCNU with calmodulin produced a concentration-dependent inhibition of in vitro calmodulin activity expressed as stimulation of cyclic nucleotide phosphodiesterase. Cyclohexylisocyanate produced a similar inhibition. Streptozotocin and chlorozotocin had no effect. Calmodulin inhibition by methyl CCNU was dependent on the concentration of calcium in the preincubation mixture. Administration of methyl-CCNU or chlorozotocin IP to CF1 mice produced a dose dependent inhibition of calmodulin activity in the small intestine. Methyl-CCNU produced a significant decrease in intestinal calmodulin activity as early as 1 h after treatment, an effect that persisted up to 52 h. Morphologic changes in the intestinal crypt epithelial cells were evident between 27 h and 5 days after treatment, but not earlier than 27 h. Renal and testicular calmodulin activity and morphology were unaffected. Although it was not possible to correlate the extent of calmodulin inhibition with severity of the intestinal lesions, the data suggested a relationship between reduced activity of calmodulin in a tissue and the ultimate appearance of lesions. This apparent interaction between an antitumor drug and calmodulin in vivo could have multiple implications for cancer chemotherapy. PMID- 2982511 TI - Tumour cell resistance to anthracyclines--a review. AB - Resistance to anthracyclines is the major factor limiting their clinical utility. Laboratory studies using cultured experimental and human tumour cells have indicated that reduced intracellular drug accumulation is one important factor underlying resistance. In some systems this results from enhanced active drug efflux, a process which may be circumvented experimentally, for example by calcium antagonists. A specific glycoprotein which is produced in excess and is inherited has been identified in the cell membrane of certain anthracycline resistant cells, while gene amplification with the appearance of double-minute chromosomes has been noted in others. Thus it is possible that anthracycline resistance arises following inherited changes in the cell membrane resulting in failure of drug accumulation. However, other possibilities exist, including differences in drug binding, either to the cell membrane or to nuclei, differences in metabolism to the semiquinone free radical, and differences in drug penetration related to tumour morphology. For each human tumour type the factor(s) involved may differ, but sufficient clues now exist to suggest that clinical testing of some of the therapeutic possibilities for circumventing anthracycline resistance may soon be appropriate. PMID- 2982512 TI - Degradation of type IV collagen by neoplastic human skin fibroblasts. AB - An assay for the degradation of type IV (basement membrane) collagen was developed as a biochemical marker for neoplastic cells from chemically transformed human skin fibroblasts. Type IV collagen was isolated from basement membrane of Syrian hamster lung and type I collagen was isolated from rat tails; the collagens were radioactively labelled by reductive alkylation. Both collagens were in the native form as shown by their low sensitivity to degradation by trypsin. The abilities of normal (KD) and chemically transformed (Hut-11A) human skin fibroblasts to degrade the collagens were studied. The normal cells released a collagenase into the medium which after activation by trypsin or oxidized glutathione degraded type I collagen. Hut-11A cells also produced a collagenolytic activity that degraded type I collagen; however, no activation of the medium was required for this activity. Type IV collagen was not degraded by medium conditioned with the normal (KD) cells with or without activation. In contrast, Hut-11A cells secreted an active collagenase into the medium that degraded type IV collagen extensively. Treatment of the medium from Hut-11A cells with trypsin resulted in only a loss of activity while treatment with oxidized glutathione was without effect. The degradation of type IV collagen by Hut-11A conditioned medium was linear for up to 1 h and the extent of degradation increased linearly with increasing amounts of conditioned medium. A cell associated assay was performed by growing either normal or transformed cells in the presence of radioactively labelled type IV collagen and measuring the released soluble peptides in the medium. This assay also demonstrated that KD cells failed to synthesize an activity capable of degrading type IV collagen whereas Hut-11A cells degraded type IV collagen in a linear manner for up to 4 h. Human serum at very low concentrations, EDTA and L-cysteine inhibited the enzyme activity, whereas protease inhibitors like phenylmethyl sulfonyl fluoride, N ethyl maleimide or soybean trypsin inhibitor did not inhibit the enzyme from Hut 11A cells. These results suggest that the ability to degrade specifically type IV collagen may be an important marker for neoplastic human fibroblasts and supports a role for this collagenase in tumor cell invasion. PMID- 2982513 TI - Role of reactive oxygen in tumor promotion: implication of superoxide anion in promotion of neoplastic transformation in JB-6 cells by TPA. AB - The role of reactive oxygen (RO) in the promotion of neoplastic transformation of JB6 mouse epidermal cells by 12-O-tetradecanoylphorbol-13-acetate (TPA) was investigated using inhibitors of RO itself or RO generating systems of seven different types. Bovine erythrocyte CuZn superoxide dismutase (SOD) maximally decreased anchorage-independent (AI) colony induction by TPA in semi-solid agar in a dose-dependent manner to 10% of TPA control level. The inhibitory effect was specifically on induction of transformation, not expression of transformation. Copper (II) (3,5-diisopropylsalicylic acid)2, which exhibits biomimetic SOD activity, was also effective. Two enzyme eliminators of H2O2, catalase and glutathione peroxidase, failed to prevent TPA-promotion. Among three hydroxyl radical scavengers, D-mannitol and Na-benzoate were moderately active but tetramethylurea did not specifically inhibit AI colony induction by TPA. A quencher of singlet oxygen, 1,4-diazobicyclo-[2,2,2]octane was also inactive. Antioxidants blocked AI transformation by TPA moderately (n-propyl gallate and tannic acid) or weakly (BHA). BHT did not specifically inhibit promotion of transformation. The effects of three inhibitors of the arachidonic acid cascade were examined. NDGA and quercetin (lipoxygenase inhibitors) were moderately active but indomethacin (cyclooxygenase inhibitor) was much less active. Based on these results, we suggest that superoxide anion (O2-.) is required for promotion of transformation by TPA. H2O2 and 1O2 appear not to be required. Hydroxyl radicals and lipid peroxides, possibly associated with O2-. action or formed in the course of oxidative metabolism of arachidonic acid also appear to be required but to a lesser extent. Products of the lipoxygenase pathway of arachidonic acid metabolism but not the cycloxygenase pathway may be important in promotion of transformation by TPA in JB6 mouse epidermal cells. The epidermal cells themselves can be both the source of and the target of the reactive oxygen in promotion. PMID- 2982514 TI - Structural and immunological identity of rat hepatoma and fetal liver nuclear poly(A) polymerase. AB - Poly(A) polymerase was partially purified from isolated nuclei of fetal rat liver. Antibodies produced in rabbits immunized with purified nuclear poly(A) polymerase from a rat hepatoma exhibited nearly identical affinity for the partially purified fetal liver and hepatoma enzymes. The extent of the antibody reaction with adult liver nuclear poly(A) polymerase partially purified in a similar manner was only 1.4% of that obtained with the hepatoma enzyme. Immune complex formation was observed between the antibodies and a major polypeptide in the fetal liver enzyme preparation which corresponded to the hepatoma enzyme (mol. wt. 48 000). No other polypeptide in the fetal liver enzyme preparation reacted with the antibodies. The 48-kDa fetal liver polypeptide produced a CNBr cleavage pattern identical to that of hepatoma poly(A) polymerase which is known to be different from the cleavage pattern of the adult liver major nuclear poly(A) polymerase. A fetal liver polypeptide corresponding to the adult liver enzyme (mol. wt. 38 000) was not evident. These results coupled with other data suggest that the hepatoma nuclear poly(A) polymerase is an oncofetal protein. PMID- 2982515 TI - Poly ADP-ribosylation and DNA strand breakage in SV40 minichromosomes. AB - Poly ADP-ribosylation and DNA strand breakage in response to treatment with the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were studied on SV40 minichromosomes in SV40-infected, permeabilized CV-1 monkey cells. After an initial sharp increase in poly ADP-ribosylation, strand breakage and poly ADPR increased proportionately with increasing dose of MNNG. This suggests a cause effect relationship between the two reactions. The major poly ADP-ribose acceptor of minichromosomes was core histone H2B. In contrast, H2B, H2A, H1 and protein A24 were poly ADP-ribosylated in the nuclear chromatin of the same cells. PMID- 2982516 TI - [125I]Aminobenzyladenosine, a new radioligand with improved specific binding to adenosine receptors in heart. AB - The density of adenosine receptors in membranes derived from rat hearts in 25 times lower than the density of receptors in rat brain membranes. Consequently, adenosine radioligands which are useful in brain such as l [3H]phenylisopropyladenosine, [3H]cyclohexyladenosine, [3H]-2-chloroadenosine and l-[125I]hydroxyphenylisopropyladenosine are of limited usefulness in heart, due to a high ratio of nonspecific to specific binding. We have synthesized a new radioligand, [125I]-N6-4-aminobenzyladenosine, which binds to rat heart membranes with one-sixth the nonspecific binding of the other radioligands. [125I]-N6-4 aminobenzyladenosine bound to rat ventricle membranes with a KD equivalent to that of l-[125I]hydroxyphenylisopropyladenosine and a Bmax of 15.2 fmol/mg protein. [125I]-N6-4-aminobenzyladenosine bound with a higher affinity to brain (KD = 1.93 nM) than to heart membranes (KD = 11.6 nM). At the radioligand KD, 60% of the total [125I]-N6-4-aminobenzyladenosine bound to heart membranes was specifically bound. Iodination of aminobenzyladenosine increased its affinity for the adenosine receptor by 22-fold, possibly due to a steric or hydrophobic effect of iodine. The new ligand was found to be a full adenosine agonist based on its ability to inhibit cyclic adenosinemonophosphate accumulation in isolated embryonic chick heart cells and rat adipocytes. [125I]-N6-4-Aminobenzyladenosine bound to a single affinity site and was displaced from cardiac and brain adenosine receptors by other adenosine analogues with a potency order of l phenylisopropyladenosine greater than 5'-N-ethylcarboxamide adenosine. These characteristics suggest that the radioligand binds to an Ri adenosine receptor. PMID- 2982517 TI - Identification and characterization of leukotriene C4 receptors in isolated rat renal glomeruli. AB - The immediate reduction of renal blood flow and glomerular filtration rate in response to intravenous infusion of leukotriene C4 in the rat prompted an analysis of isolated rat renal glomeruli for the presence of specific receptors for leukotriene C4. Specific binding of [3H]leukotriene C4 to glomeruli increased in a time-dependent manner, reached equilibrium after 60 minutes of incubation at 4 degrees C, and was 80% reversible upon addition of excess unlabeled leukotriene C4 at equilibrium. Specific binding of [3H]leukotriene C4 to glomeruli increased in a dose-dependent manner, approaching saturation at concentrations of 40-60 nM. Inhibition of binding of [3H]leukotriene C4 with increasing concentrations of unlabeled leukotriene C4 was dose dependent. The equilibrium dissociation constant for [3H]leukotriene C4 binding to glomeruli, calculated from saturation and competitive binding-inhibition studies, was 25 +/- 7 nM and 35 +/- 16 nM (mean +/- SEM), respectively, and glomerular leukotriene C4 receptor density was 8.5 +/- 1.5 and 9.0 +/- 3.0 pmol/mg protein, respectively. The other natural vasoactive sulfidopeptide leukotrienes, leukotriene D4 and leukotriene E4, the chemotactic agent, leukotriene B4, and the sulfidopeptide leukotriene antagonist, FPL 55712, competed for the receptor at concentrations 2-3 orders of magnitude higher than the homoligand, leukotriene C4. The binding and specificity characteristics of the glomerular leukotriene C4 receptor are similar to those previously reported for the DDT1 nonvascular smooth muscle cell line derived from hamster vas deferens, for guinea pig ileum smooth muscle, and for a subcellular fraction of rat lung homogenate, and represent the first characterization of such a receptor in a vascular tissue. PMID- 2982518 TI - alpha-Adrenergic receptors in cerebral microvessels of normotensive and spontaneously hypertensive rats. AB - In rat cerebral microvessels, we characterized alpha 1- and alpha 2-adrenergic receptors, using [3H]prazosin and [3H]-p-amino-clonidine as radioligands. [3H]Prazosin binding to the cerebral microvessels was saturable and of high affinity (dissociation constant of 78 pM), with a maximum binding of 48 fmol/mg protein. [3H]Prazosin binding reached equilibrium within 15 minutes and was dissociated by the addition of 10 microM phentolamine. The inhibitory effects of isomers of norepinephrine and epinephrine on the binding showed that l-isomers were over 10 times more potent than d-isomers. [3H]-p-Amino-clonidine binding to the cerebral microvessels was saturable and of high affinity (KD = 0.61 nM) with a Bmax of 73 fmol/mg protein. The binding reached equilibrium within 30 minutes, and was dissociated by the addition of 100 microM l-norepinephrine. l-Isomers of norepinephrine and epinephrine were over 10 times more potent than d-isomers in displacing the binding. Thus, both [3H]prazosin and [3H]-p-amino-clonidine bindings to the cerebral microvessels were characterized by saturability, high affinity, reversibility, and stereo-specificity. Furthermore, the specificity of both binding sites was pharmacologically evaluated by the inhibitory effects of various adrenergic agonists and antagonists on the bindings. These data indicate the existence of alpha-adrenergic receptors in the cerebral microvessels and are consistent with the hypothesis that the cerebral microcirculation is regulated by adrenergic innervation. Furthermore, the receptors were measured in cerebral microvessels of spontaneously hypertensive rats and Wistar-Kyoto controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982519 TI - Association between the exercise ejection fraction response and systolic wall stress in patients with chronic aortic insufficiency. AB - We studied the exercise ejection fraction response in 56 patients with chronic aortic insufficiency. All had left ventricular dilatation but preserved resting ejection fraction and minimal or no symptoms. The exercise ejection fraction increased by 0.05 units or greater in 18 (32%) patients (group I), remained within 0.05 units of the resting value in 18 (32%) patients (group II), and fell by 0.05 units or greater in 20 (36%) patients (group III). There were no significant differences among the groups in left ventricular end-diastolic dimension, end-systolic dimension, or fractional shortening by echocardiography or in resting left ventricular volumes and ejection fraction by radionuclide angiography. Left ventricular end-systolic wall stress was significantly higher in group III than in either group I or group II (89 +/- 20 vs 70 +/- 18 and 69 +/ 17 X 10(3) dyne/cm2; p less than .005). At peak exercise there were no differences among groups in systolic blood pressure. However, end-systolic volume increased from 65 +/- 28 to 77 +/- 36 ml/m2 in group III and fell from 50 +/- 21 to 28 +/- 18 ml/m2 in group I during exercise. Thus, at peak exercise end systolic volume was nearly three times greater in group III than in group I. Although stress could not be determined directly during exercise, the directional changes in its determinants suggest that it also would have been higher in group III patients. A highly significant inverse correlation was present between the ejection fraction response and the change in end-systolic volume (r = -.87, p less than .0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982520 TI - Assessment of myocardial ischemia with proton magnetic resonance: effects of a three hour coronary occlusion with and without reperfusion. AB - Proton (hydrogen-1) magnetic resonance imaging techniques have potential for the detection and characterization of changes associated with myocardial ischemia. Since image contrast is dependent on T1 and T2 relaxation times, we examined these parameters in a canine preparation of occlusion of the left anterior descending coronary artery. Of 16 dogs studied, seven underwent 3 hr of coronary artery occlusion and nine underwent 3 hr of occlusion followed by 1 hr of reperfusion. After the dogs were killed, the hearts of four from each group were imaged in a small bore, 1.4 tesla magnet. From all hearts myocardial segments were obtained from the normal zone and the central ischemic zone (CZ). These segments were divided into epicardial and endocardial sections and studied in a spectrometer (20 MHz). After 3 hr of occlusion, CZ endocardial T1 and T2 increased significantly (p less than .01 and p less than .05, respectively). Changes in CZ epicardial relaxation times were not as marked. Although T1 and T2 tended to be higher in the reperfused group compared with the nonreperfused group, the differences did not reach statistical significance. In keeping with the spectrometric findings, T1-dependent inversion recovery images and T2 dependent spin-echo images of the excised hearts demonstrated excellent contrast between normal and ischemic myocardium. Modest correlations were noted for both groups between blood flows during occlusion measured by the microsphere technique and T1 and T2 relaxation times. In summary, relaxation times T1 and T2 increase regionally after 3 hr of coronary artery occlusion and tend to be accentuated by reperfusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982521 TI - Elevation of brachial arterial blood velocity and volumic flow mediated by peripheral beta-adrenoreceptors in patients with borderline hypertension. AB - Simultaneous determinations of cardiac output and brachial arterial blood flow were performed in patients with hypertension and high cardiac output in comparison with normal subjects of the same age. Brachial arterial blood flow was measured with a previously described pulsed Doppler apparatus that permitted the noninvasive determination of arterial diameter and blood flow velocity. In patients with borderline hypertension, brachial blood flow was significantly increased (136 +/- 11 vs 72 +/- 8 ml/min; p less than .001). After short-term administration of indomethacin, cardiac output decreased while brachial blood flow remained constant. After short-term administration of a selective beta 1 receptor antagonist (primidolol) and nonselective blocker (propranolol), cardiac output decreased significantly in both cases but the decrease in brachial blood flow was significant only after the administration of the nonselective beta blocking agent. The study strongly suggested that in patients with borderline hypertension, the increased cardiac output is related to a prostaglandin and beta 1-adrenergic mechanisms whereas the increased brachial blood flow depends mainly on beta 2-adrenergic mechanisms. PMID- 2982522 TI - Treatment of viral myocarditis with ribavirin in an animal preparation. AB - The therapeutic effects of ribavirin, a broad-spectrum, antiviral agent, on experimental myocarditis caused by encephalomyocarditis (EMC) virus were investigated. Four-week-old DBA/2 mice were inoculated with 10 plaque-forming units (pfu) of EMC virus. Ribavirin in a dose of 100 (group 1, n = 20), 200 (group 2, n = 10), or 400 mg/kg/day (group 3, n = 10) was administered subcutaneously on days 0 to 12 after virus inoculation, and animals were observed for 12 days. Control animals were injected with saline (n = 20). Mice treated with ribavirin survived longer than controls (mean survival 6.7 days for group 1, 7.4 days for group 2, 7.7 days for group 3, and 5.2 days for control; p less than .005). Myocardial virus titers on days 6 to 8 were significantly lower in group 2 (3.24 +/- 0.49 log10pfu/mg; p less than .005) and in group 3 (1.70 +/- 0.65 log10 pfu/mg; p less than .001) compared with controls (4.09 +/- 0.57 log10 pfu/mg). The incidence of gross myocardial lesions was significantly lower in group 1 (13/20, 65%), group 2 (2/10, 20%), and group 3 (0/20, 0%) compared with controls (20/20, 100%) (p less than .05). Histologic examination showed extensive myocardial necrosis and cellular infiltration in untreated groups; there was less infiltration in groups 2 and 3 (p less than .01) and less severe necrosis in group 3 (p less than .01). Thus ribavirin effectively inhibited myocardial virus replication and reduced the inflammatory response and myocardial damage in an experimental preparation of viral myocarditis. PMID- 2982523 TI - Quantification of acylcarnitines. PMID- 2982524 TI - Natural cell-mediated cytotoxic activity against isolated chondrocytes in the mouse. AB - The ability of lymphoid cells from normal mice to exert a natural cytotoxic activity against isolated syngeneic and allogeneic epiphyseal chondrocytes was studied by means of 51Cr release assay. We found, that both spleen and peritoneal cells, but not thymocytes, exerted an anti-chondrocyte cytotoxic effect. Addition of unlabelled chondrocytes markedly reduced experimental 51Cr release and the inhibitory effect was proportional to the number of 'cold' cells added. This indicate, that chondrocyte lysis was due to specific effector-target interaction. As no cytotoxicity was observed against isolated fibroblasts, our results could not be explained by lysis of fibroblasts contaminating chondrocyte cultures. PMID- 2982525 TI - The effects of cyclosporin A on the induction, expression and regulation of the immune response to herpes simplex virus. AB - Investigations were conducted to determine the effect of cyclosporin A (CsA) on the delayed type hypersensitivity (DTH) and antibody response of mice to Herpes simplex virus (HSV). Given only at the time of priming, the drug had little effect on the subsequent DTH response in mice receiving a 'DTH immunogenic' inoculation regime. However, CsA restored normal responsiveness in groups receiving a 'DTH tolerogenic' regime implying the abrogation of T suppressor (Ts) cells. Ts cell induction was insensitive to cyclophosphamide. Antibody responses were not suppressed after giving CsA with either of these regimes and enhancement was shown in some groups. DTH was substantially reduced by CsA when the drug was given repeatedly between the time of priming and challenge, or when previously primed mice received the drug shortly before challenge. PMID- 2982526 TI - Enhanced release of reactive oxygen intermediates by immunologically activated rat Kupffer cells. AB - Release of O2- and H2O2 from isolated rat liver Kupffer cells was studied by making use of the methods of SOD sensitive ferricytochrome c reduction and horseradish peroxidase catalysed scopoletin oxidation, respectively. Kupffer cells from BCG treated rats showed a 1.8 times significantly higher O2- release and a 2.4 times higher H2O2 release as compared to the controls. Moreover the yield of Kupffer cells was also increased with administration of BCG. These results suggest that Kupffer cells can be immunologically activated to secrete larger amounts of O2- and H2O2. PMID- 2982527 TI - The clinical manifestations of a genetically determined deficiency of the third component of complement in the dog. AB - The clinical manifestations of a genetically determined deficiency of C3 were examined in a closed colony of dogs. One hundred and twelve dogs, including twenty C3-deficient dogs, were studied over a period of 6 years. Five of the C3 deficient dogs developed significant bacterial infections, such as pneumonia, sepsis, and pyometra, which were caused by Clostridium spp., Escherichia coli, and Klebsiella spp. Two of the C3-deficient dogs who had had significant infections also subsequently developed renal disease. Secondary amyloidosis was the predominant renal lesion in one dog. The predominant renal lesion in the second dog was membranoproliferative glomerulonephritis, although some amyloid was also present. The two dogs with renal disease also had positive rheumatoid factors. No other clinical or serological evidence of autoimmune disease or immune complex disease has been found. None of the dogs heterozygous for C3 deficiency, and none of the homozygous normal dogs in the colony has developed significant bacterial infections or renal disease. Thus, dogs deficient in C3, like C3-deficient humans, demonstrate both an increased susceptibility to infection and renal disease. PMID- 2982528 TI - Benign recurrent isolated VI nerve palsy of childhood. AB - Benign recurrent isolated VI nerve palsy of childhood is a rare condition. The diagnosis is essentially one of exclusion. The following case report is of a child with 11 recurrences between age 10 months and 11 years. The literature is reviewed. PMID- 2982529 TI - Platelet alpha-2 adrenoceptors and the menstrual cycle. AB - To clarify further the suggested influence of menstrual cycle phase on platelet alpha 2-adrenoceptors, we carried out cross-sectional studies in 77 subjects in a clinical trial of weight control strategies. Blood samples were drawn at baseline and after 6 weeks of diet, behavior modification, and exercise, a program that resulted in a mean weight loss of 4.5 kg. For the analyses, 42 premenopausal women were divided into four groups according to the week of menstrual cycle at the time of blood sampling. At baseline, there was no significant difference in mean platelet alpha 2-adrenoceptor numbers among the four groups. At week 6 accompanying the weight loss, there was a significant increase in the platelet alpha 2-adrenoceptor number for all groups. Despite the fact that the women were at a different phase of the menstrual cycle than at baseline, there was again no significant difference in mean platelet alpha 2-adrenoceptor number. Mean baseline platelet alpha 2-adrenoceptor number in the premenopausal women (113.7 +/- 5.5 fmol/mg protein) did not differ from values in 12 postmenopausal women (113.7 +/- 12.0 fmol/mg protein), four women with hysterectomies (105.9 +/- 8.9 fmol/mg protein), or 19 men (101.8 +/- 6.2 fmol/mg protein). Numbers at 6 weeks also did not differ. We conclude that the menstrual cycle has minimal effects on platelet alpha 2-adrenoceptor number and should not confound clinical studies of platelet alpha 2-adrenoceptors. PMID- 2982530 TI - Mechanisms of abnormalities in host defences against bacterial infection in liver disease. PMID- 2982531 TI - A comparison of cobalamin binding by liver and kidney in rat and man. AB - Binding of cobalamin (Cbl) was compared in liver and kidney plasma membranes prepared from rat and human tissues. Single, high-affinity, saturable (200 pmol/l), binding sites for TC II-Cbl were found in all tissues; by contrast no receptors were present for free cobalamin, for which only non-specific adsorption occurred. Binding constants for TC II-CNCbl determined for liver and kidney plasma membranes were of a similar magnitude. Mean values for Ka (litre/nmol) were 16.7 (rat liver), 18.8 (rat kidney), 8.0 (human liver) and 7.5 (human kidney). Results for binding TC II-OHCbl instead of TC II-CNCbl showed no difference in Ka and Bmax. values, although the non-specific adsorption was decreased to a third. Competitive inhibition results showed that the receptors are specific for the TC II molecule and that binding is unaffected either by the cobalamin moiety or by the presence of free cobalamin. Degradation of the receptor protein molecules by trypsin (10 micrograms/ml) resulted in 90% inhibition of binding. It is concluded that differences between liver and kidney in cobalamin uptake and accumulation cannot be attributed to differences in their TC II receptors. PMID- 2982532 TI - Is salt reabsorption in the human sweat duct subject to control? AB - There is a time lag between the beginning of sweat secretion and fully effective production of fluid of minimum salt content. It is suggested that changes in permeability to water or electrolytes have to be effected and may account for the time lag. The composition of thermal and pilocarpine sweat indicates that pharmacological stimulation does not wholely reproduce the physiological mechanism: pilocarpine sweat contains more sodium and more cyclic AMP, irrespective of the secretory rate. The sweat obtained from patients with cystic fibrosis has been compared with that from normal children: the concentrations of cyclic AMP and cyclic GMP are the same. Defective sodium reabsorption in the patients is therefore unlikely to be due to inadequate or excessive synthesis of these cyclic nucleotides. PMID- 2982533 TI - Effects of dietary fish oil supplementation on the fatty acid composition of the human platelet membrane: demonstration of selectivity in the incorporation of eicosapentaenoic acid into membrane phospholipid pools. AB - The fatty acid composition of membrane phospholipids of stimulated and unstimulated platelets was studied in six normal volunteers given a daily dietary supplement of a fish oil rich in eicosapentaenoic acid (EPA) for 4 weeks. The supplement was equivalent to 1.8 g of EPA daily. Thromboxane synthesis and platelet aggregation responses to sodium arachidonate, thrombin and the ionophore A23187 were also investigated. A marked increase in the relative EPA content of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) was noted after 2 and 4 weeks fish oil supplementation. However, there was no incorporation into phosphatidylinositol (PI) or phosphatidylserine (PS). The relative arachidonic acid (AA) content of PC and PE was significantly reduced at 2 and 4 weeks but that of PI and PS remained unchanged. Significant reductions in the relative linoleic acid content of total phospholipids, PC and PE were also noted. Stimulation of platelets obtained after 4 weeks fish oil supplementation by thrombin and A23187 was associated with a marked reduction in the AA content of PI and a minor reduction in that of PE. There was no change in the relative proportions of EPA in PI, PS, PC or PE after stimulation. Throughout the study there were no significant changes in platelet aggregation responses or in platelet thromboxane production. Our results indicate that the incorporation of EPA into the platelet membrane phospholipids is selective and that if PI is the major source of AA for platelet prostaglandin biosynthesis then the reported beneficial effects of EPA on haemostasis cannot be explained on the basis of its incorporation into and mobilization from the platelet membrane phospholipid pool. PMID- 2982534 TI - Is the urinary excretion of prostaglandin E in man dependent on urine pH? AB - The relationship between urine pH, modified by the oral administration of either ammonium chloride or sodium bicarbonate, and the urinary excretion of prostaglandin E (PGE) was studied in healthy female subjects. The urinary concentration of PGE, normally constant in man, was significantly higher (P less than 0.02) in alkaline than in acid urine. pH-dependent metabolism and flow dependent excretion were identified as two factors liable to obscure the demonstration of the pH-dependency of the urinary excretion rate of PGE in man. PMID- 2982535 TI - Proceedings of a workshop on alpha-adrenoceptors. Loch Lomond, 27-28 July 1984. PMID- 2982536 TI - alpha 2-Adrenoceptors in dog kidney: autoradiographic localization and putative functions. AB - The location of alpha 2-adrenoceptors has been examined in slide-mounted sections of dog kidney by using labelling in vitro with [3H]rauwolscine and autoradiography. [3H]Rauwolscine binding in dog kidney was of high affinity, saturable, and to a single population of sites. Stereoselectivity was observed for (-)-noradrenaline and the relative affinity of a series of competing antagonists indicated binding to alpha 2-adrenoceptors. Autoradiography of [3H]rauwolscine binding revealed localization to three areas: glomeruli, the intimal and adventitial surfaces of arcuate blood vessels and the medullary vascular bundles. PMID- 2982537 TI - The adrenaline-alpha 2-adrenoceptor-mediated vasoconstrictor axis. AB - In patients with essential hypertension plasma adrenaline concentrations have been found to be higher than in normotensive subjects and this may represent increased adrenergic activity. Adrenaline released into the circulation can be taken up by the sympathetic nerve ending and as it is re-released as a co transmitter it enhances exocytotic noradrenaline release by stimulating prejunctional beta-adrenoceptors and as a consequence it contributes to postjunctional alpha 1-adrenoceptor-induced vasoconstriction. Adrenaline may also induce vasoconstriction via post- and extra-junctional alpha 2-adrenoceptors, as shown by a decrease in the forearm blood flow during adrenaline infusions in the postjunctional alpha 1- and beta-blocked forearm vasculature, an effect that could be antagonized by alpha 2-adrenoceptor blockade with yohimbine. alpha 2 Adrenoceptor stimulation in platelets showed an increased sensitivity to adrenaline, as determined by sensitivity in counteracting the inhibitor effect of PGI2 on intracellular free calcium concentration in untreated patients with essential hypertension, when compared with treated patients or normotensive subjects. As these effects can be normalized by antihypertensive treatment this suggests that the increased hormone sensitivity may be related to the elevated intracellular free calcium concentration. Thus adrenaline, via pre- and post junctional adrenoceptors, may contribute to enhanced vascular smooth muscle contraction, which most likely is sensitized by the elevated intracellular calcium concentration. PMID- 2982538 TI - Vascular alpha-adrenoceptors in man: interactions with adrenaline and noradrenaline. AB - Adrenaline and noradrenaline were infused in cumulative doses into the brachial artery of six healthy volunteers and changes in forearm blood flow (FBF) were measured by plethysmography. The catecholamines were infused together with saline, yohimbine and doxazosin, in that order. All infusions were given in the presence of intra-arterial propranolol to prevent beta-adrenoceptor-mediated effects. Adrenaline and noradrenaline reduced FBF significantly. This decrease in FBF was significantly attenuated by doxazosin as well as by yohimbine. No differences were found between adrenaline and noradrenaline. It is concluded that postjunctional alpha 1- and alpha 2-adrenoceptors are involved in the vasoconstriction caused by exogenous adrenaline and noradrenaline in man. The relevance of circulating adrenaline for the alpha 2-adrenoceptor-mediated contribution to basal vascular tone remains to be established. PMID- 2982539 TI - Factors determining alpha-adrenoceptor-mediated responses of human digital arteries and metacarpal veins. PMID- 2982540 TI - Role of alpha-adrenoceptors in vascular control. PMID- 2982542 TI - Pain control in dentistry: the basis for rational therapy. PMID- 2982541 TI - Enalapril, a nonsulfhydryl angiotensin-converting enzyme inhibitor. AB - The chemistry, pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosage of enalapril maleate, a nonsulfhydryl angiotensin-converting enzyme (ACE) inhibitor, are reviewed. Enalapril is rapidly converted by ester hydrolysis to enalaprilat, a potent ACE inhibitor; enalapril itself is only a weak ACE inhibitor. Enalapril lowers peripheral vascular resistance without causing an increase in heart rate. In patients with congestive heart failure, enalapril has beneficial hemodynamic effects based on reduction of both cardiac preload and afterload. Approximately 60% of a dose of enalapril is absorbed after oral administration. Excretion of enalaprilat is primarily renal. Accumulation of enalaprilat occurs in patients with creatinine clearances less than 30 mL/min. Enalapril 10-40 mg per day orally has shown efficacy comparable to that of captopril in treating patients with mild, moderate, and severe hypertension, hypertension caused by renal-artery stenosis, and in congestive heart failure resistant to digitalis and diuretics. When given alone for hypertension, enalapril has efficacy comparable to that of thiazide diuretics and beta blockers. Side effects observed with enalapril have generally been minor. Captopril-associated side effects such as skin rash, loss of taste, and proteinuria have been observed in a small number of patients receiving enalapril to date; neutropenia less than 300/mm3 has been noted with captopril but not enalapril. The incidence of these side effects has been noted to be greatly decreased in patients on low doses of captopril. Enalapril appears to be similar in efficacy to captopril for treating hypertension and congestive heart failure. Whether enalapril is safer than low-dose captopril in patients at high risk for captopril-associated side effects will require further investigation. PMID- 2982543 TI - Understanding dental pain: Part I. PMID- 2982544 TI - Quantitative analysis of polymorphonuclear leukocyte superoxide anion generation in critically ill children. AB - Apart from its well-known bactericidal action, superoxide anion produced by activated polymorphonuclear leukocytes is believed to be involved in pathophysiology of diffuse capillary leak syndromes, e.g., adult respiratory distress syndrome and septic shock. Neutrophil NADPH oxidoreductase, the superoxide anion synthetase, was assayed in blood samples from a variety of critically ill children. Neutrophils from patients with evidence of diffuse capillary leak syndrome had significantly depressed enzyme specific activity as compared to neutrophils from healthy controls or intensive care patients without evidence of diffuse capillary leak. These data along with additional in vitro findings support the contention that previously activated granulocytes may be refractory to subsequent activation, and that such behavior may represent an intrinsic defense mechanism to minimize inflammatory amplification autoinjury. PMID- 2982545 TI - Examination of the enzyme assay for NADPH oxidoreductase; application to polymorphonuclear leukocyte superoxide anion generation. AB - Superoxide anion production in intact polymorphonuclear leukocytes isolated from medically stable pediatric ICU patients was investigated by examining the initial velocity enzyme kinetics of NADPH oxidoreductase, under a variety of experimental conditions. Present findings confirm that enzyme activity requires stimulation of the respiratory burst, and that catalysis followed longitudinally reflects activation, activity, and termination phases. There was no superoxide anion production in unstimulated neutrophils, and only short-lived superoxide generation in neutrophils stimulated by phorbol myristate acetate. Moreover, the ability to activate resting neutrophils was a labile phenomenon, as reflected by an inducible enzyme half-life of 1.2 days. There were no apparent differences in enzyme activity from granulocytes isolated from venous vs. arterial blood. Enzyme activity in intact granulocytes peaked at pH 7 and 30 degrees C, where the ratio of enzyme activity to cell number was constant over the range 0.25 to 2.5 X 10(5) cells per assay. Utilizing the above strictly defined assay conditions, pediatric derived polymorphonuclear leukocytes induced with phorbol myristate acetate were found to generate superoxide anion at rates equivalent to those in healthy adults. PMID- 2982546 TI - Adenocarcinoma of the urachus: computed tomography diagnosis. AB - Serial abdominal computed tomography scans demonstrated late bladder wall involvement by a calcified mucinous urachal adenocarcinoma that was initially confined to a chronically inflamed urachal cyst. PMID- 2982547 TI - Arachidonic acid and leukotrienes in clinical dermatology. PMID- 2982549 TI - Epidermodysplasia verruciformis. PMID- 2982548 TI - Retinoids in oncology. PMID- 2982550 TI - Transferrin receptors: structure and function. PMID- 2982551 TI - Radiotherapy alone for patients with operable carcinoma of the lung. AB - Between 1966 and 1980, 72 patients with operable carcinoma of the lung were treated primarily with radiotherapy because coexisting, nontumor related medical problem, or patient refusal prevented thoracotomy. We compared results obtained in this group with results obtained by thoracotomy in 123 consecutive patients over the age of 70, who were assessed in a similar fashion, but who underwent thoracotomy. All patients in both groups had proven, nonsmall cell carcinoma of the lung without clinical, laboratory, or radiologic evidence of tumor spread. All patients in both groups had a negative staging mediastinoscopy, and bronchoscopic findings consistent with operability. Many of the patients treated with radiotherapy had less than a curative dose as their general medical condition prevented a course of radical radiotherapy. It is apparent, however, that the results of radiotherapy for patients with operable carcinoma of the lung may be disappointing and that for patients who are marginal in terms of operative risk, the benefits of surgical resection may warrant the risks involved. PMID- 2982553 TI - Two-dimensional echocardiographic abnormalities of right atrial metastatic tumors in hepatoma. AB - We describe two patients suffering from hepatoma who presented with right atrial metastatic tumors as a result of invasion of the inferior vena cava and extension into the right atrium. Two-dimensional echocardiographic studies detected the right atrial tumor during life in both patients and the invasion of the inferior vena cava in one patient. PMID- 2982552 TI - Trial of vindesine plus mitomycin in stage-3 non-small cell lung cancer. An active regimen for outpatient treatment. AB - Ninety patients with stage-3 non-small cell lung cancer were given vindesine (3 mg/sq m) plus mitomycin (10 mg/sq m). Data on response are available for 84 adequately treated individuals (93 percent). For patients who had received no prior chemotherapy, the rate of major objective response was 36 percent (20/55). For previously treated patients the rate of major response was 17 percent (5/29). The drugs were routinely administered in the outpatient department without difficulty. Moderate or severe myelosuppression, neurotoxic, nephrotoxic, or pulmonary toxic effects, nausea, and vomiting occurred in less than 15 percent of all studied patients. Three-drug extravasation ulcerations occurred in 1,129 administrations of chemotherapy (0.3 percent). There were two treatment-related deaths, one from sepsis and one from the combination of mitomycin-induced pulmonary and renal toxic effects. The combination of vindesine plus mitomycin is an active, well-tolerated outpatient regimen for patients with non-small cell lung cancer who have not previously received chemotherapy. Further trials are warranted to compare this regimen to other active combinations and to use it as a component of a program of treatment using alternating regimens of chemotherapy. PMID- 2982554 TI - [Cystic pancreas neoplasias]. AB - For the treatment of cystic changes of the pancreas, it is essential to distinguish cysts and pseudocysts from neoplasm. Since clinical parameters are usually not characteristic, only morphologic analysis will prove a diagnosis. Nowadays, the cystic pancreatic neoplasms are described as: microcystic adenoma (rich-in-glycogene cystadenoma), and mucinous cystic neoplasms (cystadenocarcinoma or cystadenoma). Morphology, etiology, clinical findings, and treatment are discussed on 3 cases. The new classification of cystic pancreatic neoplasms is important for prognosis in two aspects: 1) On the clinical finding of a pseudocyst, every surgeon should think of a mucinous cystic neoplasm and look for solid tumours digitally. 2) For the pathologist, any mucinous neoplasm should cause him to analyse such cysts macroscopically and, if possible, also microscopically, to rule out an adenocarcinoma. PMID- 2982555 TI - Prognostic correlations of operable carcinoma of the rectum. AB - Various histologic factors correlated to survival were studied in 124 patients radically operated on for rectal carcinoma in order to establish valid prognostic criteria. The total survival rate after five years was 63 percent, while in stage B1 it was 89 percent, in B2, 61 percent, and in C1, 47 percent (P less than 0.05). With regard to histotype, the survival was 83 percent in the papillary subtype of adenocarcinoma, while in the tubular subtype it was 62 percent, and 29 percent in the mucinous type (P = not significant). Vascular invasion negatively affected survival (41 percent); however, when there was no invasion, the prognosis was better (71 percent) (P less than 0.01). In evaluating histologic grading and lymphoglandular reactivity, the difference in survival rates was not statistically significant. The marked peri- and intratumoral lymphocytic infiltration gave a very good prognosis (92 percent) contrary to when reactivity was moderate (59 percent) or even absent (51 percent) (P less than 0.01). Finally, the expanding type tumor, with reference to Ming's classification of gastric carcinoma, had a much better prognosis (75 per cent) than the infiltrative type (40 percent) (P less than 0.01). PMID- 2982556 TI - Differential effects of dietary fibers on rat intestinal circular muscle cell size. AB - The relationship between dietary fiber and intestinal circular muscle cell size was investigated in rats by feeding defined diets supplemented with four different sources of fiber. In the first study, a 20% wheat bran supplement was fed to 10 rats for nine weeks. This resulted in larger muscle cell size, with a 22.5% increase in the proximal (P less than 0.02) and 77.9% increase in the distal colon (P less than 0.01) when compared with a control group of 10 rats fed a fiber-free diet. In the second study, which lasted four weeks, a control group of 10 rats was fed a fiber-free diet, while similar sized experimental groups were fed the same basal diet plus either 20% oat bran, 10% pectin, or 10% guar. Muscle cell size was decreased by 20.6% in the proximal jejunum of the oat bran- and pectin-fed groups (P less than 0.05) and by 43% in the proximal colon of the oat bran-fed group, when compared with the controls (P less than 0.05). These results show that the effects of high fiber diets on intestinal muscle cell size depend on the type of fiber consumed. PMID- 2982557 TI - Use of avian retroviral-bovine growth hormone DNA recombinants to direct expression of biologically active growth hormone by cultured fibroblasts. AB - A variety of recombinant DNA molecules were constructed in which an avian retroviral long terminal repeat (LTR) was ligated to the bovine growth hormone (bGH) gene. The retroviral LTR was derived from a plasmid clone of a Schmidt Ruppin B strain of Rous sarcoma virus while the bGH gene was subcloned from a lambda bacteriophage genomic library. Using a transient eukaryotic expression assay system, recombinant plasmid constructs were screened for their ability to direct expression and secretion of bGH. One such plasmid DNA construct, termed pBGH-4, was found to be active in the production of bGH. Stable mouse fibroblast cell lines were generated containing pBGH-4 DNA integrated into the mouse cell genome. Many of these mouse cell lines express and secrete bGH. One line, L-Pd lambda-BGH4-13, was found to secrete bGH at a rate of 75 micrograms per 5 X 10(6) cells per 24 hr. Bovine growth hormone derived from this cell line is biologically active. PMID- 2982558 TI - A new method for purifying lambda DNA from phage lysates. AB - A new method for preparing small quantities of lambda DNA from phage lysates has been developed. The protocol is based on the concentration and purification of bacteriophage particles from crude lysates using small DEAE-cellulose columns. This chromatographic step gives an absolute separation of the lambda DNA from the cellular nucleic acids and a 20-fold enrichment relative to the major soluble proteins in crude lysates, while effecting a 10-fold concentration of the phage. Final deproteinization and concentration of the lambda DNA is achieved by conventional precipitation steps. The lambda DNA produced by this method is shown to be nondegraded, biologically active, and an excellent substrate for restriction enzymes. A detailed protocol is provided for starting with individual plaques and using the method to obtain purified DNA from large numbers of lambda clones. PMID- 2982559 TI - [Detection and intracellular localization of 2 polyphosphate phosphohydrolases with different substrate specificities in the slime variant of Neurospora crassa]. PMID- 2982560 TI - [Changes in the reaction of the heart to posthypoxic reoxygenation with the administration of superoxide dismutase and its inhibitor]. PMID- 2982561 TI - [Poisoning with hypochlorite-containing sanitary cleansers]. PMID- 2982562 TI - [Acquired immune defect syndrome: current status. Position of the German Association for the Control of Virus Diseases and the Virology Section of the German Society for Hygiene and Microbiology]. PMID- 2982563 TI - [Therapy of stomach ulcer with low-dose antacid gel and cimetidine. A multicenter double-blind study]. AB - In a multi-centre double-blind (double-dummy) trial the effectiveness of low-dose antacid gel (6 X 12 ml/d; neutralisation capacity 120 mmol) was compared with that of a standard dose of cimetidine (1 g/d) in the curative treatment of gastric ulcer. Antacid gel was given to 65 patients, cimetidine to 60. Diagnosis was confirmed by endoscopic biopsy, which was also employed in a serial follow up. After 4 weeks antacid gel and cimetidine produced cures in 43% and 52%, respectively; after 8 weeks 76% and 89%, respectively, the difference between the two methods not being statistically significant. There was also no statistically significant difference with regard to ulcer pain. In one case each in the antacid and cimetidine groups, the treatment had to be stopped because of side effects. Diarrhoea was more common on cimetidine than on antacid gel. It is concluded that both low-dose antacids gels and cimetidine are suitable in the treatment of gastric ulcers. PMID- 2982564 TI - Multiple sclerosis. Current concepts in management. AB - Multiple sclerosis is a chronic, often progressive disease of the central nervous system which can produce visual, sensory, motor, and genitourinary dysfunction. Although there is no cure, many disabling symptoms can be ameliorated. Baclofen is the treatment of choice for spasticity and is usually given in doses of 30 to 80 mg/day, although higher doses may be used. Bladder symptoms in multiple sclerosis generally fall into the categories of failure to store, failure to empty, and mixed types. Most patients can be managed after obtaining a urine culture and sensitivity and post-voiding residual. A variety of anticholinergic agents plus intermittent self-catheterisation is usually the most effective treatment for bladder dysfunction. Prevention of infection is accomplished by urinary acidifiers or low-dose antibiotics. There is no evidence that long term use of corticosteroids has a beneficial effect on the outcome of multiple sclerosis, although they appear to be useful in hastening the recovery time from an acute exacerbation. There are a number of experimental therapeutic agents which are used to modulate the immune response, which may prove to be of use in slowing or arresting the progression of multiple sclerosis. PMID- 2982565 TI - Glomus tympanicum presenting as Meniere's disease. PMID- 2982566 TI - Enhancement of rat growth hormone binding by membrane disulfide reduction. AB - The effect of disulfide reduction on the binding of [125I]rat GH (rGH) to rat liver plasma membranes and hepatocytes was studied to determine the role of disulfide bonds in the binding of GH to its receptor. The total amount of [125I] rGH bound to the liver receptors increased severalfold in the presence of dithiothreitol and mercaptoethanol. The nonspecific binding also increased at higher concentrations of the reductant, but the amount specifically bound was still greater in the presence of disulfide reductant. In contrast, the disulfide reductant inhibited [125I] human GH (hGH) binding and enhanced its displacement from hypophysectomized female rat hepatocytes. This was similar to the effect of reductants on [125I]hGH binding to normal female rat hepatocytes. The effect of the disulfide reductants on [125I]rGH binding could be prevented or reversed by the simultaneous or subsequent addition of an oxidizing agent such as NAD or oxidized glutathione. Sulfhydryl-reactive agents such as iodoacetamide prevented additional binding of [125I]rGH when added at 30 min of the incubation. The additional [125I] rGH bound in the presence of disulfide reductant was displaceable by excess unlabeled rGH. Both rGH and hGH exhibited similar degrees of disulfide reduction in the presence of mercaptoethanol. The disulfide reductant produced effects on binding at concentrations that resulted in less than 10% reduction of the GH disulfides. We conclude that: 1) the disulfides and sulfhydryls of the hepatocyte membrane are intimately involved in the binding of GH to hepatic receptors; 2) the locus of the disulfides and sulfhydryls may be in the subunit structure of the membrane receptor, but this will require verification using soluble receptors; and 3) the effect of disulfide reducing agents reveals basic differences in the mechanism of binding of rGH and hGH to somatotropic hormone receptors on the hepatocytes. PMID- 2982567 TI - Evaluation of an in vitro parathyroid hormone antagonist in vivo in dogs. AB - A potent competitive inhibitor of PTH-stimulated biological responses in vitro, [Nle8,Nle18,Tyr34] bovine PTH (bPTH)-(3-34)amide, was evaluated in vivo in dogs. These studies confirm observations in vitro, suggesting that positions 1 and 2 of the peptide are critical to its biological activity. However, unlike the results from studies in vitro, this PTH analog is a weak agonist with effects on parathyroid target tissues that produce hypercalcemia and phosphaturia and increase urinary cAMP excretion. Assessed by these three parameters of hormonal action in vivo, the estimated potency of this analog is less than 1% of that of the intact hormone. In addition, PTH-induced biological responses were not inhibited by relatively large doses of the bPTH-(3-34) analog. These results emphasize the need for a systemic, integrated approach, combining chemical with biological studies, to design effective inhibitors of hormonal action in vivo. Although the rationale for introducing particular modifications into the peptide structure is most frequently based on bioassays performed in vitro, the success of the strategy chosen must rely, ultimately, upon the demonstration of specific biological properties in vivo. PMID- 2982568 TI - Morphological and functional characterization of interstitial cells from mouse testes fractionated on Percoll density gradients. AB - Cell fractions were obtained by separation on Percoll density gradients after dissociation of mature mouse testes by mechanical or collagenase dispersion, and the ultrastructure, hCG receptor properties, and hCG-stimulated testosterone (T) production of these fractions were compared. Gradients were fractionated according to specific gravity, and all cell types were quantitated using morphometric techniques. Three peaks of specific [125I]iodo-hCG binding corresponding to densities of 1.0667 g/cm3 (fractions 2-3), 1.045 g/cm3 (fractions 6-7), and 1.0365-1.0215 g/cm3 (fraction 9) were obtained after collagenase dispersion, but the second peak of binding (fractions 6-7) was not observed after mechanical dispersion. Morphometric studies were performed by light microscopy on the cells present in the fractions corresponding to the first and second hCG binding peaks obtained by either mechanical or collagenase dispersion; the third representing germ cells and membranous debris was not studied further. Regardless of the method of preparation, morphologically intact Leydig cells represented 60-80% and 7-10% of the cells in fractions 2-3 and 6-7 that were associated with the first and second peaks of hCG binding Leydig cells obtained from fractions 6-7 contained greater numbers of lipid inclusions than Leydig cells from fractions 2-3. Mechanically dispersed Leydig cells exhibited similar numbers of hCG receptors in the dense and light Leydig cells, but hCG stimulated T production per Leydig cell was significantly greater in the dense Leydig cells containing few lipid inclusions. T production by dense and light collagenase-dispersed Leydig cells was not significantly different. The second hCG binding peak in collagenase-dispersed cell fractions 6-7 was associated with an increase in the number of an indeterminate connective cell type released from the testes by collagenase treatment, whose ultrastructure and limited hCG-binding capacity suggested that they may represent Leydig cell precursors. It is concluded that the identification and quantitation of different cell types in isolated testicular cell fractions is, therefore, of fundamental importance in the interpretation of receptor and secretory capacities of enriched preparations of Leydig cells. PMID- 2982569 TI - Pharmacological characterization of the angiotensin receptor negatively coupled with adenylate cyclase in rat anterior pituitary gland. AB - Angiotensin II (AII) inhibited anterior pituitary adenylate cyclase. Whereas GTP was necessary to fully express the AII inhibitory effect, Na+ was not required. The magnitude of inhibition (42 +/- 6%) permitted a pharmacological characterization of the AII receptor involved in adenylate cyclase inhibition. Angiotensin I (AI) was less potent than AII, and deletion of aminoacids in the N terminal position resulted in a progressive reduction of the Ki (peptide concentration producing half-maximal inhibition). The Ki values were 3 +/- 0.9, 10, and 700 nM for AII, angiotensin III (AIII), and des-Asp, des-Arg-AII, respectively. Sarcosine in position 1 [( Sar, Phe]AII) increased the potency of inhibition (Ki = 0.12 +/- 0.12 nM). Different antagonists of the AII receptors appeared to be partial agonists. There was a very close correlation (r = 0.98) between the respective potencies of a series of AII analogs to inhibit adenylate cyclase and the potencies of these analogs to elicit PRL or ACTH release or to bind to AII-binding sites. Dopamine and AII inhibition of anterior pituitary adenylate cyclase were not additive. This suggests that both receptors are on the same cell and likely on lactotrophs. This hypothesis agrees with the observation that vasoactive intestinal peptide stimulation of adenylate cyclase was inhibited by AII, whereas corticotropin-releasing factor stimulation was unaffected. Although dopamine and AII inhibited the same adenylate cyclase, they had opposing effects on PRL release (inhibition and stimulation, respectively). The possible significance of this observation is related to a model implying that PRL release can be elicited through either a Ca+2 or a cAMP pathway. PMID- 2982570 TI - Impaired parathyroid hormone receptor-adenylate cyclase system in the postobstructed canine kidney. AB - The present studies examine the initial events in the action of PTH, namely receptor binding and adenylate cyclase activation in the postobstructed canine kidney. Both kidneys were removed from five mongrel dogs 2 h after relief of unilateral ureteral obstruction (UUO) of 42 h duration. Basolateral membranes (BLM) were prepared from both the UUO and contralateral (control) kidneys. Maximal activation of adenylate cyclase by PTH was 35% lower (P less than 0.01) in BLM from UUO than control kidneys (3869 +/- 200 vs. 5978 +/- 425 pmol cAMP/mg protein X 30 min). The concentration of PTH required for half-maximal activation of the enzyme was unchanged. Addition of 1 mM GTP failed to correct the decreased enzyme activity in response to PTH of BLM from kidneys with UUO. NaF activation, a measure of interaction of the nucleotide regulatory component (G/F) with the catalytic unit of the adenylate cyclase was similar in BLM from UUO and control kidneys. Similarly, activation of the catalytic unit of the enzyme by Mn2+ was not different. Specific binding of [125I]Nle8, Nle8, Tyr34-bovine PTH NH2 was markedly reduced (P less than 0.01) in BLM from UUO vs. control (6.37 +/- 1.20 vs. 2.43 +/- 0.09 fmole bound/microgram protein). There was no change in hormone affinity for the binding site. These data indicate that there is decreased activation of adenylate cyclase by PTH as a consequence of apparent loss of receptors for the hormone in the BLMs of renal tubular cells of postobstructed kidneys. These abnormalities may play a role in the abnormal regulation of phosphorus excretion after ureteral obstruction. PMID- 2982571 TI - Hormonal regulation of myometrial estrogen, progesterone, and oxytocin receptors in the pregnant and pseudopregnant hamster. AB - Estrogen receptor (Re) and progesterone receptor (Rp) concentrations were measured in the myometrium of hamster uterus during pregnancy and pseudopregnancy. Comparison of Re and Rp levels with serum estradiol and progesterone titers revealed that receptor concentration was low when progesterone was elevated during pregnancy and pseudopregnancy. However, Re and Rp levels increased when progesterone levels dropped at the end of each condition. In comparing serum estradiol relative to progesterone at the end of pregnancy and pseudopregnancy, it was discovered that Re and Rp recovery occurred not only when the estradiol to progesterone ratio increased (pseudopregnancy) but also when the ratio did not change (pregnancy). This suggested that serum progesterone was the primary determinant of receptor down-regulation, and this was confirmed by comparing the receptor recovery response to estrogen and progesterone withdrawal in the decidualized hamster uterus. Total Re levels increased to the same extent after progesterone withdrawal whether or not serum estradiol was maintained. When serum estrogen was maintained at a steady state, nuclear Re (nRe) increased within 4 h of progesterone withdrawal, and estrogen dependent protein responses (Rp and oxytocin receptor) were obtained within 8 h. Thus, progesterone-induced down-regulation of nRe and estrogen-dependent proteins is rapidly reversed upon removal of hormone. The recovery response of Re, Rp, and oxytocin receptor to progesterone withdrawal can be blocked by cycloheximide treatment at 4 h, suggesting that receptor recovery involves protein synthesis. These results are consistent with the hypothesis that progesterone down-regulates the Re system by a selective action on nRe retention. PMID- 2982572 TI - Production of insulin-like growth factor I and its binding protein by adult rat hepatocytes in primary culture. AB - This study examines the production and characteristics of insulin-like growth factor I (IGF I) and its binding protein (BP) from adult rat hepatocytes in primary culture. IGF I and BP in samples of conditioned medium were separated by automated high pressure gel permeation chromatography at pH 2.8, with total recovery of both species. Measured by a specific RIA, with about 2% IGF II cross reactivity, the rate of production of IGF I [compared with a human IGF I (hIGF I) standard] was 4.39 +/- 0.57 pmol/mg cell protein X 24 h; measured by a radioligand assay using radioiodinated hIGF I as tracer, BP was produced at a rate of 2.97 +/- 0.54 pmol binding sites/mg cell protein X 24 h. High pressure permeation chromatography studies indicated approximate mol wts of 7,500 for hepatocyte IGF I and 50,000 for BP. The pI of hepatocyte IGF I was 8.8, and of the predominant BP species, 6.8. Rat serum treated in the same way as hepatocyte medium showed IGF I and BP activities at identical pI values to those from hepatocytes but, in addition, contained BP activity at more acidic pI values, apparently not produced directly by the liver. Competitive binding studies using acid-chromatographed hepatocyte or serum BP, and hIGF I and IGF II as labeled and unlabeled ligands, also suggested that BP activity in serum was heterogeneous, while BP from hepatocytes appeared to be a single species with similar affinities for IGF I and IGF II. It is concluded that adult rat hepatocytes produce immunoreactive IGF I, pI 8.8, and BP, pI 6.8, which are released into the circulation, but that additional, more acidic, BP species which are also found in serum could result from extrahepatic production or processing. PMID- 2982573 TI - Rat hepatocyte insulin-like growth factor I and binding protein: effect of growth hormone in vitro and in vivo. AB - This study examines the regulation of insulin-like growth factor I (IGF I) and binding protein (BP) release by adult rat hepatocytes in primary culture. Increasing the density of plating of cells had a marked positive effect on the IGF I production rate per mg cell protein, with the highest rate, approximately 4 pmol/mg cell protein X 24 h, seen at densities of 1 X 10(5) cells/cm2 or higher. Cycloheximide (15 micrograms/ml) inhibited both IGF I and BP production by more than 90%, while actinomycin D (0.1 microgram/ml) caused less marked, but still significant, inhibition. Over the insulin range 30 pM-300 nM there was a 45% increase in IGF I production, but no effect on BP production. Bovine GH stimulated production of both peptides, significant effects (up to 50% stimulation) being seen at concentrations from 20-500 ng/ml. Cells from hypophysectomized rats, with serum IGF I and GH levels reduced more than 90% from normal, had IGF I and BP production rates only 7% of normal, measured after 48 h in culture. In vivo replacement with rat GH, 150 micrograms/day for 5 days by osmotic minipump, significantly restored the production of both peptides by hepatocytes. Cells isolated from rats bearing the GH-secreting tumor, MtT/W15, had IGF I and BP production rates approximately twice as high as normal. A significant stimulatory effect of bovine GH (200 ng/ml) in vitro was seen on cells from normal and hypophysectomized, GH-replaced rats, but not from unreplaced hypophysectomized, or tumor-bearing rats. The results in hypophysectomized, animals are consistent with known changes in hepatic GH receptors, while the lack of GH responsiveness in tumor-bearing rats indicates a persistence of maximal stimulation in culture. The reflection of in vivo GH status by hepatocytes cultured for 48 h suggests that messenger RNA turnover for IGF I and BP must be slow. The close parallel in the regulation of the two peptides under most conditions might be indicative of coordinated synthesis. PMID- 2982574 TI - The acute secretory response to alterations in extracellular calcium concentration and dopamine in perifused bovine parathyroid cells. AB - We used perifusion of dispersed bovine parathyroid cells to examine the rapid kinetics of the secretory response to alterations in extracellular calcium and to dopamine and compared changes in hormone secretion with alterations in the putative intracellular mediators cytosolic calcium and cAMP. The increase in hormone secretion associated with a reduction in extracellular calcium from 2.0 to 0.75 mM occurred at least as rapidly as the change in calcium concentration, suggesting that the lag time for the stimulation of secretion was of the order of seconds or less. In cells loaded with the intracellular calcium-sensitive dye QUIN 2, on the other hand, the initial activation of hormone secretion by low extracellular calcium was delayed by 30-40 sec. In both QUIN 2-loaded and unloaded parathyroid cells, a subsequent increase in extracellular calcium concentration from 0.75 to 2.0 mM produced a rapid inhibition of hormone secretion which could not be separated temporally from the changes in extracellular calcium. In QUIN 2-loaded cells, the reduction in cytosolic calcium concentration associated with a decrease in extracellular calcium from 2.0 to 0.75 mM took place with a half-time of about 15 sec. The increase in cytosolic calcium concentration on raising extracellular calcium from 0.75 to 2.0 mM had a half-time of approximately 20 sec. Dopamine (10(-5) M) also produced a nearly immediate 3- to 4-fold stimulation of PTH release. Although the increase in hormone secretion preceded the release of cAMP from perifused cells, intracellular cAMP increased 3.4-fold within 10 sec of exposure to dopamine in parallel experiments. Preincubation of perifused cells with dopamine reduced the subsequent secretory response to low extracellular calcium. Conversely, dopamine stimulated secretion was significantly greater in cells preincubated with 2.0 than in those incubated with 0.75 mM calcium. These results indicate that perifused bovine parathyroid cells respond to secretagogues with a time course comparable to that observed in vivo and that the temporal changes in cytosolic calcium concentration and cellular cAMP are consistent with a mediatory role for these factors in low calcium- and dopamine-stimulated secretion, respectively. In addition, at least a portion of the secretory response to low calcium and dopamine may come from a common cellular pool of PTH. PMID- 2982575 TI - Possible role of cortisol in the stimulation of cortisol-binding capacity in the plasma of fetal sheep. AB - In sheep, the prepartum rise in fetal plasma cortisol (F) is associated with an increase in the corticosteroid-binding capacity (CBC) of the plasma. We examined whether CBC changed during preterm labor, induced by administering ACTH to the fetus, and we examined the role of F in such changes. Beginning on day 127, chronically catheterized fetal sheep were divided into six groups; 1) saline control infusions (n = 4); 2) pulsatile ACTH (P-ACTH; 66 ng/min for 15 min every 2 h; n = 5); 3) P-ACTH (as 2) plus metopirone (MET; 500 mg/24 h n = 5); 4) P-ACTH and MET (as 3) plus F (13.3 micrograms/min for 15 min every 2 h; n = 3); 5) P ACTH and MET (as 3) plus F (133 micrograms/min for 15 min every 2 h; n = 4); 6) P ACTH and MET (as 3) plus dexamethasone (0.56 micrograms/min for 15 min every 2 h for 50 h, then 5.6 micrograms/min for 15 min every 2 h until 100 h; n = 4). All infusions continued for 100 h. Fetal blood samples were collected at 8-h intervals for determination of plasma F concentration and CBC. Estrone and progesterone were measured in 8-h samples of maternal femoral arterial blood. P ACTH resulted in a significant increase in CBC within 24 h; values at 72-96 h were 2- to 3-fold greater than controls. Rises in CBC were positively correlated with the changes in plasma F in the P-ACTH-treated fetuses and preceded changes in plasma progesterone or estrogens. MET treatment prevented the P-ACTH-induced increases in F and CBC. Exogenous F and dexamethasone overcame the MET-induced inhibition of P-ACTH-induced increases in CBC. We conclude that in fetal sheep 1) CBC is elevated by P-ACTH treatment; and 2) F may mediate this stimulation of its own binding protein. PMID- 2982576 TI - Growth hormone binding to specific receptors stimulates growth and function of cloned insulin-producing rat insulinoma RIN-5AH cells. AB - Binding of 125I-labeled human GH (hGH) to a cloned rat insulin-producing cell line RIN-5AH in monolayer culture was studied along with some physiological effects of the hormone on these cells. Binding was time and temperature dependent, and steady state binding was observed in 60 min at 37 C with [125I]hGH at 4.2 pM, whereas at 24 C, binding had not reached a steady state after 120 min. The binding was largely reversible, since 80% of initially bound [125I]hGH dissociated from the cells upon incubation in hGH-free buffer for 120 min. Half maximal binding was obtained when cells were incubated in the presence of 3.0 X 10(-10) M unlabeled hGH. Rat GH as well as human placental lactogen were able to compete for binding sites, but with less affinity. Other non-GH peptides at 6.7 micrograms/ml did not affect [125I]hGH binding. Scatchard analysis revealed curvilinear plots, and approximately 2700 high affinity binding sites were calculated. Culture of RIN-5AH in the presence of 1 microgram/ml hGH for 4 days resulted in an 80% increase in insulin content as well as an 18% increase in cell number and DNA and protein content compared to those in cells cultured in the absence of hGH. The dose dependence of the insulinotropic effect showed that half maximal and maximal stimulation were observed in cells cultured in the presence of 10 and 100 ng/ml, respectively. Insulin release to the medium during the 4-day culture period was not affected by hGH. These data suggest that GH, through binding to specific receptors in the cell membrane, directly stimulates proliferation and function of pancreatic beta-cells. PMID- 2982577 TI - Induction of flavokinase (EC 2.7.1.26) by aldosterone in the rat kidney. AB - The effect of aldosterone on rat renal flavokinase (EC 2.7.1.26) enzymic activity and concentration was investigated in bilaterally adrenalectomized male Sprague Dawley rats. Flavokinase enzymic activity was measured in the 100,000 X g supernatant of renal cortex and red medulla and was increased after 3 h by 19% and 42%, respectively, as a result of aldosterone (1.5 micrograms/100 g BW) administration. Dual isotope labeling studies revealed increases of 20-30% and 20 35% in the incorporation of [3H]- and [35S]methionine into rat renal cortical and red medullary flavokinase, respectively, as a result of aldosterone administration. The aldosterone-dependent increases in methionine incorporation were blocked when the mineralocorticoid receptor antagonist spirolactone (SC 26304; 150 micrograms/100 BW) was administered 30 min before aldosterone. The relative concentrations of renal flavokinase also increased by 40% in both the cortex and red medulla 3 h after aldosterone treatment, as determined by an enzyme-linked immunosorbent assay (ELISA). These increases were abolished by actinomycin D (100 micrograms/100 g BW) and cycloheximide (200 micrograms/100 g BW), given 1 h before aldosterone administration. The mineralocorticoid receptor antagonists SC 26304 (225 micrograms/100 g BW) and progesterone (500 micrograms/100 g BW) were also able to inhibit the aldosterone-dependent increase in renal flavokinase concentration. The effect of aldosterone on renal flavokinase concentration was studied from 30 min to 8 h after aldosterone administration. There appeared to be maximum increases of 23-30% and 25-32% after 2.5-3.5 h in the renal cortex and red medulla, respectively. 5 alpha Dihydroaldosterone, a metabolite of aldosterone with mineralocorticoid activity, was also able to increase renal flavokinase concentrations by approximately 40%. However, dexamethasone, a potent glucocorticoid with little or no mineralocorticoid activity, appeared to have no effect on renal flavokinase, as observed by ELISA. These data suggest that the increase in the relative concentration of renal flavokinase may be due to increased biosynthesis of flavokinase, and ultimately, that renal flavokinase may be an aldosterone-induced protein whose synthesis is mediated through the mineralocorticoid receptor and RNA synthesis. PMID- 2982578 TI - Characterization of cell surface adenosine 3',5'-monophosphate-binding proteins in Y-1 mouse adrenal tumor cells. AB - Adrenal cortical cells are known to export cAMP and have binding proteins and cAMP-dependent protein kinase activity associated with their plasma membranes. Because these properties suggest a function for extracellular cAMP, we have undertaken a search for specific cell surface receptors for this cyclic nucleotide. Y-1 mouse adrenal tumor cells actively export cAMP by an energy dependent process. Analysis of Scatchard plots of the equilibrium binding of [3H]cAMP to these cells indicate the existence of two classes of cAMP binders: one with high affinity (ka = 2.9 X 10(9) M-1) and another with low affinity (ka = 7.0 X 10(7) M-1). The cell surface localization of these binders was established by the sensitivity of both the [3H]cAMP-binding proteins and the [32P]8-N3-cAMP photoaffinity labeled proteins of intact cells to mild trypsin digestion and by the surface distribution of a BSA-O2-monosuccinyl cAMP-gold complex revealed by electron microscopy. Analysis of radioautograms of cell surface cAMP-binding proteins from confluent monolayer tumor cells, photoaffinity labeled with [32P]8 N3-cAMP and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two major 32P-labeled protein bands which were indistinguishable from the 49,000 and 55,000 mol wt regulatory subunits of the cytosolic protein kinase isoenzymes of this cell. These observations along with the demonstration of cell surface, cAMP-dependent protein kinase activity in the mouse adrenal tumor cell strongly suggest that these cAMP-binding proteins function as regulatory proteins for cell surface protein kinases. PMID- 2982579 TI - Inhibition by lysosomotropic amines of dog thyroid secretion in vitro. AB - The lysosomotropic amines are well-known inhibitors of lysosomal protein degradation. These drugs were used in dog thyroid slices to ascertain the role of the lysosome in thyroglobulin hydrolysis and in hormone secretion. NH4Cl (1-20 mM) and chloroquine (5-500 microM) inhibited, in a concentration-dependent manner, the TSH-stimulated secretion. The high concentrations of these compounds also inhibited the basal secretion. The inhibition was not toxic since 1) the nonbutanol extractable iodine, an index of follicle disruption, was not increased, except slightly by 20 mM NH4Cl; 2) the compounds did not inhibit the TSH-induced stimulation of protein iodination; 3) the inhibition of secretion caused by these compounds was reversible; 4) the ultrastructural changes induced by NH4Cl were reversed after its withdrawal. The inhibition of secretion was presumably related to the lysosomal trapping of the drugs because: 1) the time lag for the fall in secretion rate after drug addition was shorter than for the inhibition of secretion at the level of pseudopod formation by carbamylcholine and cytochalasin B; 2) for NH4Cl this delay and the degree of inhibition were modulated by the [H+] gradient between the medium and the lysosome; 3) 20 min after NH4Cl addition, 92% of the lysosomes were vacuolated and swollen (median section area, 126 mu2 vs. 50 mu2 for the controls) and 8% of the lysosomes were swollen and still dense (median, 206 mu2); 20 min after chloroquine addition, 90% of the lysosomes remained dense and had a significantly higher section area (median, 79 mu2) than the controls (P less than 0.001), whereas 10% of the lysosomes were vacuolated and large (median area, 438 mu2). The number of pseudopods measured by scanning electron microscopy significantly decreased only after 1 h (P less than 0.001). This late decrease could not account for the early block of secretion and suggested a lack of membrane recycling. In conclusion, lysosomotropic amines interfere with a post phagocytotic, presumably lysosomal, step in secretion by dog thyroid. These data constitute the first biochemical evidence in the intact cell of the role of lysosomes in TSH-induced thyroglobulin hydrolysis and thyroid hormone secretion. PMID- 2982580 TI - Stress-induced secretion of adrenocorticotropin in rats is inhibited by administration of antisera to ovine corticotropin-releasing factor and vasopressin. AB - Intact handled rats were pretreated with the immunoglobulin G fractions from normal rabbit serum or antisera to ovine corticotropin-releasing factor (CRF) and/or vasopressin and subjected to restraint or formalin stress. The formalin induced rise in plasma ACTH was reduced to 28% in rats pretreated with anti-CRF, to 53% in those pretreated with antivasopressin, and to 16% in rats given both antibodies. Pretreatment of animals with anti-CRF, antivasopressin, or a combination of both antibodies also attenuated the ACTH response to restraint stress to 13%, 37%, and 12%, respectively, of those in normal rabbit serum treated rats. Antiserum pretreatment did not reduce the restraint- or formalin induced rise in plasma PRL in the same animals, however. We conclude, therefore, that both vasopressin and an ovine CRF-like peptide are physiologically relevant peptides involved in stress-induced ACTH release. PMID- 2982581 TI - Regulation of the 1,25-dihydroxyvitamin D3 receptor by 1,25-dihydroxyvitamin D3 in intact human cancer cells. AB - Human breast cancer cells have been shown to be targets of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]action with specific dose-dependent effects on growth in vitro. These cells possess specific, high affinity receptors for 1,25-(OH)2D3. We have studied the interaction of the vitamin D hormone with its receptor in these intact human target cells. Specific binding of tritium-labeled 1,25-(OH)2D3 by these cells reaches a maximum with 3-9 h depending on hormone concentration; subsequently there is a rapid loss of hormone-binding activity to virtually undetectable levels by 12-24 h. Although the hormone induces its own metabolism in these cells, binding cannot be restored by the addition of fresh hormone. This loss of hormone-binding activity of the receptor, is analogous to the processing of the estrogen receptor in estrogen receptor-positive cells. Two inhibitors of DNA transcription, actinomycin D and cordycepin, delay the onset and slow the processing of receptor if added with the hormone. They are ineffective if added after hormone binding is maximal. Inhibitors of protein synthesis have no protective effect at all and, in fact, these agents inhibit both the attainment of maximum specific binding and the recovery of receptor over the subsequent 24 72 h. These data demonstrate for the first time that the 1,25-(OH)2D3 receptor undergoes regulation in the intact human target cell. This event, which resembles processing of the estrogen receptor and down-regulation of peptide-hormone receptors, likely represents a nuclear mechanism for the control of cellular responsiveness to hormone in the intact cell. PMID- 2982582 TI - Fasting induces the generation of serum thyronine-binding globulin in Zucker rats. AB - Five-month-old lean and obese Zucker rats were fasted for up to 7 days (lean rats) or 28 days (obese rats), and serum total and free T4 and T3 concentrations, percent free T4 and T3 by equilibrium dialysis, and the binding of [125I] T4 to serum proteins by gel electrophoresis were measured. In the lean rats, a 4- or 7 day fast resulted in significant decreases in serum total and free T4 and T3 concentrations. There was a decrease in the percent free T3 after 7 days of starvation. In contrast, a 4- or 7-day fast did not alter any of these variables in the obese rats. However, after 14 or more days of starvation, serum total T4 and T3 concentrations increased, and the percent free T4 and T3 decreased, resulting in no change in the serum free T4 or T3 concentrations in the obese rats. The percent of [125I]T4 bound to serum thyronine-binding globulin increased and the percent bound to thyronine-binding prealbumin decreased with the duration of the fast in both the lean and obese rats. The increase in serum thyronine binding globulin binding of T4 can explain the increase in serum total T4 and T3 concentrations, the decrease in percent free T4 and T3, and the normal free hormone concentration in the long term fasted obese rats. The findings in the lean rats appear to be due to a combination of the known central hypothyroidism that occurs during 4-7 days of fasting and the fasting-induced changes in T4 binding in serum. Changes in T4 and T3 binding in serum during fasting in the rat must be considered when the effects of fasting on serum concentrations of the thyroid hormones, thyroid hormone kinetics, and the peripheral action of the thyroid hormones are evaluated. PMID- 2982583 TI - Ovine placental lactogen inhibits glucagon-induced glycogenolysis in fetal rat hepatocytes. AB - The effect of ovine placental lactogen (oPL) on glucagon-stimulated glycogenolysis was studied in cultured hepatocytes from 20-day-old fetal rats. Pretreatment of hepatocytes with oPL (0.5-5 micrograms/ml) significantly attenuated the inhibitory effect of glucagon on glycogen synthesis. Hepatocytes exposed to glucagon alone at 1, 5, and 20 nM incorporated 32.0, 43.2, and 62.1% less [14C]glucose into glycogen than control hepatocytes. However, hepatocytes pretreated for 1 h with oPL (1 microgram/ml) and then exposed to the same concentration of glucagon incorporated only 5.8, 9.2, and 22.1% less [14C]glucose than control cells (P less than 0.01 vs. glucagon alone). In cells preincubated for 24 h in medium containing [14C]glucose, glucagon reduced cellular [14C]glycogen and total glycogen content by 47.1 and 51.0% while oPL increased total cellular glycogen content by 105.8% and attenuated the glycogen-degradative effect of glucagon. While oPL alone had no effect on basal phosphorylase a (Pa) activity, oPL (1-10 micrograms/ml) caused a 15.3-91.6% inhibition of glucagon stimulated Pa activity (P less than 0.01). The maximal inhibition by oPL of glucagon-stimulated Pa activity occurred within 2 min of exposure to oPL, and the effect was not blocked by cycloheximide. oPL also caused a 49.4-95.0% inhibition of (Bu)2cAMP-stimulated Pa activity (P less than 0.01), suggesting that the inhibitory effect of oPL on glucagon action is exerted, at least in part, at a site distal to the intracellular accumulation of cAMP. Insulin (1 microM) reduced basal Pa activity, abolished the stimulation of Pa activity by glucagon, and markedly attenuated the stimulation of Pa by (Bu)2-cAMP. These studies demonstrate that oPL acutely inhibits glycogen degradation in fetal rat hepatocytes and suggest that oPL promotes glycogen storage in fetal liver both by antagonizing the glycogenolytic effects of glucagon and by stimulating fetal hepatic glycogenesis. PMID- 2982584 TI - A comparison of the mechanisms of alpha-adrenergic inhibition of thyrotropin stimulated adenosine 3',5'-monophosphate in cat, rat, mouse, hamster, beef, and pig tissues with the stimulatory effect of epinephrine on beef thyroid iodination: evidence for multiple, species-specific adrenergic mechanisms. AB - Epinephrine was shown to inhibit TSH-stimulated cAMP formation in cat and pig thyroid slices and isolated rat and hamster thyroid lobes. In contrast, no such inhibitory action could be demonstrated in sheep or beef thyroid slices or mouse thyroid-trachea preparations. The inhibitory effect of epinephrine on TSH stimulated cAMP formation in pig thyroid slices was blocked by 10 microM yohimbine, but not by 10 microM prazosin, suggestive of mediation through an alpha 2-catecholamine receptor mechanism. In cat thyroid slices, the inhibitory effect of epinephrine was blocked by both yohimbine and prazosin, suggestive of a mixed alpha-adrenoceptor mechanism. Meclofenamate and indomethacin attenuated the epinephrine response in cat, but not pig, thyroid slices, but other prostaglandin inhibitors were ineffective. Beef thyroid slices responded to epinephrine with an increase in iodide organification that is mediated through an alpha-adrenergic mechanism not blocked by propranolol. This stimulatory effect of epinephrine was abolished by both 10 microM prazosin and 10 microM yohimbine. In contrast to the inhibitory effect of the catecholamines on TSH-stimulated cAMP formation in cats, the stimulatory response of beef iodide organification to epinephrine was not modified by meclofenamate, indomethacin, or verapamil. These findings suggest that the receptor mechanisms mediating the inhibitory effect of catecholamines on cat thyroid and the stimulatory effect of catecholamines in beef thyroid slices may well be mediated by separate and as yet undefined receptor mechanisms. In contrast, the inhibitory effect of epinephrine on TSH-stimulated cAMP formation in pigs is most likely mediated through an alpha 2-adrenoceptor mechanism. These findings further document the multiplicity of catecholamine actions on thyroid function as well as the diversity observed among various species. PMID- 2982585 TI - Radiation inactivation (target size analysis) of the gonadotropin-releasing hormone receptor: evidence for a high molecular weight complex. AB - In the present study we used radiation inactivation (target size analysis) to measure the functional mol wt of the GnRH receptor while it is still a component of the plasma membrane. This technique is based on the observation that an inverse relationship exists between the dose-dependent inactivation of a macromolecule by ionizing radiation and the size of that macromolecule. This method demonstrates a mol wt of 136,346 +/- 8,120 for the GnRH receptor. This estimate is approximately twice that obtained (60,000) by photoaffinity labeling with a radioactive GnRH analog followed by electrophoresis under denaturing conditions and, accordingly, presents the possibility that the functional receptor consists of a high mol wt complex in its native state. The present studies indicate that the GnRH receptor is either a single weight class of protein or several closely related weight classes, such as might occur due to protein glycosylation. PMID- 2982586 TI - Analysis of hormone- and polynucleotide/histone-binding sites of the chicken intestinal 1,25-dihydroxyvitamin D3 receptor by means of proteolysis. AB - Several discrete forms of the 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] receptor from chicken intestinal cytosol were characterized by ultracentrifugation; gel filtration; DNA-, histone-, dye-ligand-binding affinity; and several other chromatographic media. Formation of altered receptor complexes was carried out through partial proteolysis using trypsin, alpha-chymotrypsin, and papain. The holoreceptor complex was found to be asymmetric and have a Stoke's radius of 37 A. Depending on the concentration of protease, several sterol-binding fragments of 33 and 27 A were produced which tended to be more globular in shape. The 27 A complex was incapable of binding DNA, histones, or phosphocellulose, but still retained affinity for DEAE-cellulose. Resolution of three forms of 1,25-(OH)2D3 receptors was demonstrated by cibacron blue F3GA-agarose chromatography. The 37 A complex eluted at 1.1 M KCl, and the 33 A form eluted at 0.6 M KCl, whereas the 27 A complex was not retained by the column and eluted in the 0.15 M KCl wash. The specificity of the 37 A complex for histone binding was assessed. The order of binding preference was: histone f3 greater than histone f2a approximately equal to histone f2b much greater than histone f1. Results also indicated that DNA could inhibit receptor binding to histone and that this was competitive with respect to histone-agarose binding, suggesting that the interaction of histones and DNA is at a domain common to polynucleotides. It is concluded the 1,25 (OH)2D3 receptor has separate and distinct binding sites for hormone and polynucleotides/histones. These in vitro findings of histone binding suggest that the polynucleotide domain of this receptor is capable of recognizing several nuclear derived components that may be important for the alteration of gene expression. PMID- 2982587 TI - Stress hormones in blood and cerebrospinal fluid of conscious sheep: effect of hemorrhage. AB - Acute moderate hemorrhage (15 ml/kg withdrawn over 10 min) was used to study stress hormone changes in blood and cerebrospinal fluid (CSF) of conscious sheep with chronic indwelling intracerebroventricular catheters. Mean plasma arginine vasopressin (AVP) and ACTH rose 150- and 14-fold, respectively, above basal values to peak levels at 20 min after onset of hemorrhage. A smaller (4- to 5 fold) rise occurred in plasma angiotensin II (AII) to peak levels at 10 min. The corticosteroid response (cortisol and aldosterone) occurred later (peak at 45 min) and was consistent with the dependence of these steroids on plasma ACTH and AII changes. Increases in plasma epinephrine and norepinephrine were small and transient. Compared to changes in plasma, changes in CSF hormone levels after hemorrhage were small and independent of plasma concentrations. Mean CSF AVP increased to peak levels at 15 min whereas rises in CSF ACTH, AII-like immunoreactivity, and cortisol were slower and delayed in comparison with the patterns observed in plasma. Despite evidence of increased sympathetic activity, and rise in plasma catecholamines, CSF epinephrine fell after hemorrhage and CSF norepinephrine did not change. These results show that in conscious sheep rapid and major increases in plasma AVP, ACTH, and AII follow acute moderate hemorrhage. Concomitant changes in CSF hormone levels are small and delayed. With the possible exception of AVP it appears unlikely that the acute systemic hormone response to hemorrhage is determined by hormone changes in CSF. PMID- 2982588 TI - Modulation of differentiated function in cultured thyroid cells: thyrotropin control of thyroid peroxidase activity. AB - The activity of thyroid peroxidase (TPO) in primary dog thyroid cell cultures was measured by both guaiacol oxidation and iodide oxidation assays. Whether cultures were initiated in the absence or presence of 50 mU/ml TSH, TPO activity fell in the first 24 h of culture to approximately 10% of the activity in freshly isolated follicles. After 5 days in culture, TPO activity almost completely disappeared in the absence of TSH, whereas in the presence of TSH, TPO activity rebounded to approximately 30% of that in freshly isolated follicles. TSH similarly induced TPO activity in cells that had lost this activity during a 1- to 6-day preincubation period in the absence of hormone. The half-time for the induction of TPO activity was approximately 3 days. Whether TSH was present from the start of culture or added after 5 days of culture without TSH, the half maximal dose for reinduction of TPO activity was 0.3-0.4 mU TSH/ml. (Bu)2cAMP, 8 bromo-cAMP, forskolin, and cholera toxin all mimicked, either completely or in part, the ability of TSH to induce TPO activity in cells preincubated without hormone. We conclude that, in cultured dog thyroid cells, TPO activity is modulated by chronic TSH stimulation, and that this effect is mediated by cAMP. However, even though TSH stimulates TPO activity in cultured thyroid cells, the fact that there is no comparable restoration of organic iodine formation (as found in previous studies) makes it likely that other aspects of the iodide organification mechanism are altered. PMID- 2982589 TI - Adenylate cyclase in the corpus luteum of the rhesus monkey. II. Sensitivity to nucleotides, gonadotropins, catecholamines, and nonhormonal activators. AB - The sensitivity of the adenylate cyclase of the primate corpus luteum to various nucleotides, gonadotropins, catecholamines, and nonhormonal activators was assessed in homogenates of luteal tissue obtained from rhesus monkeys at the midluteal phase of the menstrual cycle. The conversion of [alpha-32P]ATP to [32P]cAMP was used to monitor adenylate cyclase activity. GTP, the GTP analog 5' guanylyl-imidodiphosphate, and ITP stimulated adenylate cyclase activity in the presence or absence of exogenous hormone; however CTP, UTP, GMP, and guanosine did not. The gonadotropins, human (h) LH and hCG, stimulated cAMP production in a dose-dependent manner. Maximal stimulation of adenylate cyclase was achieved at 100 nM hLH and hCG, and the activation constant was 20 nM for both hormones. The addition of GTP increased maximal activation of adenylate cyclase by hLH or hCG, but did not alter sensitivity to the hormones. Neither hFSH nor the isolated subunits of hCG stimulated cAMP production. Deglycosylated hCG (native hCG with 70% of the carbohydrate moieties removed) did not stimulate adenylate cyclase activity. However, hLH and intact hCG failed to enhance cAMP production in the presence of an equimolar amount of deglycosylated hCG. The adenylate cyclase of macaque luteal tissue did not respond to the addition of isoproterenol, epinephrine, or phenylephrine. Furthermore, these catecholamines did not affect hCG stimulation of adenylate cyclase. The nonhormonal activators of adenylate cyclase, forskolin and fluoride, stimulated cAMP production in a dose-dependent manner, with maximal stimulation at 100 microM and 10 mM, respectively. Thus, the macaque corpus luteum at the midluteal phase of the menstrual cycle contains a guanine nucleotide-regulated adenylate cyclase which is equally sensitive to the pituitary and placental gonadotropins, hLH and hCG. However, removal of carbohydrate moieties from hCG endows the molecule with gonadotropin-antagonistic properties in the primate. The adenylate cyclase system of the macaque corpus luteum was not responsive to catecholamines; thus, the primate may lack a potential mechanism for control of luteal function that is available to many nonprimate species. PMID- 2982590 TI - Ontogeny of hypothalamic luteinizing hormone-releasing hormone (GnRH) and pituitary GnRH receptors in fetal and neonatal rats. AB - Although it is known that LH secretion starts at 17 days of gestation in the fetal rat and that this first LH release is most likely driven by hypothalamic GnRH, an earlier role for GnRH during fetal life has been postulated with the observation that presence of GnRH is important before day 13 of gestation for the differentiation of the pituitary anlage. In order to clarify the different roles of GnRH during fetal life, we have studied the first appearance of GnRH in the fetal brain, the expression of GnRH receptors in the fetal pituitary gland, and the presence of GnRH immunoreactivity within the fetal gonadotrophs. GnRH was present in the earliest brain tissue examined (12 days of gestation). From 12-17 days, GnRH content of fetal brain remained low and then increased markedly by the end of gestation. No immunoreactive GnRH-like material could be detected in rat placental tissue throughout gestation. Binding sites for GnRH were detected as early as 12 days of gestation in fetal pituitary glands. However, binding was very low until 16 days. At 17 days, Scatchard analysis indicated the presence of high affinity, low capacity binding sites [affinity constant (Ka) = 10(10) M-1]. Intracellular presence of GnRH as seen by immunocytochemistry using ultrathin sections prepared by cryoultramicrotomy was first visible at 14 days and started to increase at 16 days. LH was first detectable in the fetal pituitary by RIA at 17 days; FSH was first detectable at 21 days, and PRL at 1 day of postnatal life. Thereafter, neonatal pituitary contents of LH, FSH, and PRL increased linearly with-time, as did the number of pituitary GnRH receptors. At 10 days of postnatal life, pituitary contents of LH and FSH were significantly higher in females than in males. In summary, hypothalamic GnRH appears early in fetal life and potentially can induce differentiation of the pituitary anlage. Conversely, the presence at 15 days of gestation of specific binding sites for GnRH and of intracellular GnRH immunoreactivity in gonadotrophs indicates that the hypophysiotropic action of GnRH clearly precedes the start of LH biosynthesis. PMID- 2982592 TI - Phorbol esters stimulate growth and inhibit differentiation in cultured thyroid cells. AB - The potent tumor promoter 12-O-tetradecanoylphorbol 13-acetate (TPA) has biological effects on cell growth and differentiation similar to the effects of epidermal growth factor (EGF) on a variety of cells. Since EGF has been shown recently to stimulate thyroid cell proliferation and inhibit iodine metabolism, we examined the effects of phorbol esters on primary ovine thyroid cultures. TPA stimulated cell growth in a manner similar to EGF. The growth effects of EGF and TPA in combination were not additive. In contrast, TPA (1.6 X 10(-7) M) was a more potent inhibitor of iodine uptake and incorporation than EGF (10(-9) M) at their maximally effective concentrations. The inhibitory effects of TPA were also more rapid and less reversible than those of EGF. TPA and EGF in combination inhibited iodine metabolism more than either agent alone at its maximally effective concentration. Both TPA and EGF reduced the accumulation of cAMP in TSH stimulated cells, but (Bu)2cAMP and stimulators of adenylate cyclase failed to overcome TPA's inhibition of iodine metabolism. TPA interacted with EGF by reducing the affinity of membrane receptors for [125I]iodo-EGF. Although the alteration in EGF-receptor interaction induced by TPA may play a role in mediating TPA's biological effects, the additive effects of TPA and EGF on iodine metabolism suggest that TPA does not act solely through the EGF receptor-effector system. Agents other than TSH, including phorbol esters and EGF, are potent modulators of thyroid growth and differentiated function. Despite several similarities in biological activity, TPA and EGF do not modulate differentiated function in an identical manner. Both factors act at least partially through a non-cAMP-dependent pathway, providing indirect evidence of another second messenger(s) in the control of thyroid function. PMID- 2982591 TI - Differential plasma beta-endorphin, beta-lipotropin, and adrenocorticotropin responses to stress in rats. AB - This study was designed to compare the amounts of ACTH, beta-endorphin (beta END), and beta-lipotropin (beta LPH) that are present in plasma under basal conditions and after single and repeated administration of a discrete 2-min restraint stress both in intact and in chronically adrenalectomized rats. In intact rats, application of a 2-min restraint stress produced rapid parallel increases in plasma concentrations of radioimmunoassayable ACTH and beta END/beta LPH (the total of beta END-like immunoreactivities), with peaks 2.5-5 min after onset of the stress and return almost to basal concentrations by 30 min. Gel exclusion chromatography [Sephadex G-50 (fine)] and subsequent RIA revealed that plasma obtained from control nonstressed intact rats contained much greater quantities of beta END (94% of the total beta END/beta LPH immunoreactivity) than beta LPH. In contrast, equal amounts of beta END and beta LPH were present in plasma of intact rats 2.5-10 min after onset of the 2-min restraint stress. Chronically adrenalectomized rats lacking glucocorticoid-negative feedback had significantly higher basal plasma concentrations of beta END/beta LPH and ACTH than those present in intact rats. Furthermore, the plasma responses of both beta END/beta LPH and ACTH to stress were markedly enhanced in chronically adrenalectomized rats compared to the corresponding responses in intact rats. Gel exclusion chromatography revealed that both the higher basal concentration and the enhanced plasma beta END/beta LPH response to stress in adrenalectomized rats resulted primarily from increases in the beta LPH component, with lesser increases in the beta END component. In contrast to the proportion in intact rats, in chronically adrenalectomized rats, beta END represented about 27% of the total beta END/beta LPH immunoreactivity in the basal state and about 18% 5-10 min after the onset of restraint stress. In intact rats, the plasma ACTH responses to a subsequent stress applied 5 min (a time when peak plasma levels of hormones are present) or 30 min (a time when the plasma hormone concentrations have returned to prestress levels) after the initial stress and the plasma beta END/beta LPH response to a second stress applied at 30 min were equal to the corresponding hormone responses to a single stress. In contrast, the plasma beta END/beta LPH response to the subsequent stress applied 5 min after the initial stress was significantly potentiated in intact rats.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982593 TI - Lactogen receptors in rat Leydig cells: analysis of their structure with bifunctional cross-linking reagents. AB - [125I]Iodohuman GH was found to bind to receptors with specificity for lactogenic hormones in a Triton X-100 extract from Leydig cell membranes displaying an affinity constant of 3.8 X 10(9) M-1 and a binding capacity of 167 fmol/mg protein. Cross-linking of solubilized [125I]iodohuman GH-receptor complexes with disuccinimidyl suberate followed by analysis by sodium dodecyl sulfate-gel electrophoresis in the presence of beta-mercaptoethanol and autoradiography resulted in the appearance of bands with apparent mol wt of 113,000, 103,000, 59,000, and 53,000. The appearance of these bands was prevented by incubation in the presence of lactogenic hormones. By using a two-dimensional electrophoresis technique (first dimension under nonreducing conditions; second dimension under reducing conditions), it was demonstrated that a fraction of the mol wt 59,000 species can be released from the mol wt 103,000 species upon cleavage of disulfide bonds. These results suggest the existence of lactogen receptor species with approximate mol wt of 91,000, 81,000, 37,000, and 31,000 in Triton X-100 extracts from Leydig cell membranes if the contribution of the free hormone (mol wt, 22,000) is subtracted. A fraction of the mol wt 37,000 subunits appears to be contained within the 81,000 species linked through disulfide bonds. PMID- 2982594 TI - Regulation of corticotropin-releasing factor (CRF) receptors in the rat pituitary gland: effects of adrenalectomy on CRF receptors and corticotroph responses. AB - The stimulation of ACTH release from anterior pituitary cells by corticotropin releasing factor (CRF) is mediated by specific, high affinity receptors with a Ka of 10(9) M-1 for ovine CRF. The relationship between ACTH secretion and CRF receptor activation was analyzed in normal and adrenalectomized rats by comparison of ACTH release with changes in CRF receptors and adenylate cyclase activity. The marked increase in plasma ACTH levels that occurred after adrenalectomy (from 71 to 478 pg/ml after 4 days) was accompanied by a progressive decrease in pituitary CRF receptor concentration [by 29 +/- 1%, 75 +/ 2%, 77 +/- 6%, and 80 +/- 4% (+/- SE) after 1, 2, 3, and 4 days, respectively]. Most of this decrease was due to receptor down-regulation rather than occupancy by endogenous CRF, since high dose infusions of CRF (300-500 ng/min) for 30 min before pituitary membrane preparation reduced CRF-binding sites by only 40%. The marked reduction in CRF receptors after adrenalectomy was accompanied by comparable decreases in maximal CRF-stimulated adenylate cyclase activity and sensitivity to CRF (ED50, 3.8 +/- 2.8 vs. 58 +/- 3.7 X 10-9 M CRF in control and 2-day-adrenalectomized rats, respectively). Fluoride-stimulated adenylate cyclase activity was unchanged at 24 h, but was decreased by 28 +/- 7% at later times. Such decreases in CRF receptors and adenylate cyclase activity in adrenalectomized rats were prevented by dexamethasone treatment. In cultured anterior pituitary cells from 4-day-adrenalectomized rats, CRF-stimulated cAMP production was decreased by 40%. However, in contrast to the decreases in CRF receptors and cAMP production, there was a 3-fold increase in CRF-stimulated ACTH release, with no change in sensitivity to CRF. The ability of corticotrophs to maintain increased ACTH release, in conjunction with reduced CRF receptors and CRF-stimulated adenylate cyclase, indicates that elevated ACTH secretion can be maintained by occupancy and activation of only a small number of CRF receptors. This finding also suggests that synergistic interactions between CRF and other regulators of ACTH release may contribute to the sustained increase in ACTH secretion that follows adrenalectomy. PMID- 2982595 TI - Receptor mediated action without receptor occupancy. AB - The relationship between occupancy of a specific cell surface receptor and actions initiated in the same cell by that event was tested on ovarian granulosa cells exposed to hCG. Hormone bound to its receptor was identified by immunocytochemistry or by autoradiography and the dissociation of cAMP-dependent protein kinase was followed by direct cytochemistry. Only 25-30% of the granulosa cells bound hCG and in each instance this resulted in protein kinase dissociation. Cells that did not bind hCG nevertheless dissociated protein kinase if they contacted a cell that had bound hormone and dissociated enzyme. Cells that neither bound hormone, nor contacted cells that did so, failed to dissociate protein kinase. These observations establish, in individual cells, a direct relationship of receptor occupancy to receptor-mediated action and indicate that this event can be communicated to receptorless cells, presumably by gap junctions, thereby amplifying the response to hormone. Similar processes may occur in other tissues wherein receptor-bearing cells are capable of intercellular communication. PMID- 2982596 TI - Effects of acute and chronic phenytoin on the electrolyte content and the activities of Na+, K+-, Ca2+, Mg2+-, and HCO3- -ATPases and carbonic anhydrase of neonatal and adult rat cerebral cortex. AB - Various parameters of anion and cation transport were measured in the cerebral cortex of neonatal (3-day-old) and adult rats following acute and chronic treatment with phenytoin (PHT). Acutely, PHT significantly inhibited the enzyme Na+, K+-ATPase in both neonatal and adult rats. This effect was accompanied by a significant increase in cerebral cortical Na+ content and a decrease in K+ content only in neonatal animals. Chronic treatment (two and four times a day for 7 days) of adult rats with PHT significantly reduced Na+ content without affecting whole homogenate Na+, K+-ATPase activity. The activity of this enzyme was markedly increased in the myelin- (glial product) and slightly decreased in the synaptosomal- (neuronal) fractions following chronic (four times a day for 7 days) PHT treatment. These results suggest that PHT differentially affects the two forms (neuronal and glial) of the enzyme Na+, K+-ATPase. The possible relevance of this hypothesis in relationship to the anticonvulsant and excitatory properties of PHT is discussed. Chronic (two and four times a day for 7 days) PHT treatment increased both DNA content and activity of the glial marker enzyme carbonic anhydrase. Activity of the mitochondrial enzyme HCO3- -ATPase was also increased following chronic PHT treatment. These two enzymes are intimately involved in the regulation of HCO3- -Cl- transport across glial cell and mitochondrial membranes, and these results suggest that PHT is able to affect beneficially glial regulatory processes. The ability to enhance glial regulation of anions and cations in extracellular fluid provides new and important insights into the mechanism of the anticonvulsant action of PHT. PMID- 2982597 TI - Effects of phenytoin on primary glial cell cultures. AB - The activity of enzymes involved in anion and cation transport, the concentration of intracellular potassium (K+i), and the transmembrane potential (Em) were determined following acute and chronic exposure of primary astroglial cultures to micromolar concentrations of phenytoin (PHT). Na+, K+-ATPase activity of homogenates of cultured glial cells was determined in the presence of an increasing K+ concentration (1-20 mM). Acutely, PHT had little effect on the K+ activation pattern of Na+, K+-ATPase. In contrast, the percentage of Na+, K+ ATPase activated by elevating the K+ concentration was dose dependently increased by chronic PHT treatment. This effect was accompanied by a marked increase in K+i and a significant membrane hyperpolarization. The acute effect of PHT on the Em was biphasic, characterized by membrane hyperpolarization at concentrations of 10(-6)-10(-5) M; at concentrations between 10(-5) and 10(-4) M, the Em progressively returned to control values. These results suggest that glial cells acutely and chronically treated with therapeutic concentrations of PHT have an enhanced capacity to control elevated extracellular potassium levels. Return of the Em to control values at PHT concentrations greater than 10(-5) M suggests that these cells are less able to regulate extracellular potassium. These data can partially explain the excitatory effects of PHT at high therapeutic concentrations. PMID- 2982598 TI - A type-II DNA topoisomerase and a catenating protein from the transplantable VX2 carcinoma. AB - It has recently been suggested that topoisomerases could be important targets for several DNA intercalating drugs used in cancer therapy. This prompted us to purify and characterize a type II topoisomerase in a highly tumorigenic transplantable rabbit tumor isolated from a skin carcinoma associated with cottontail rabbit papillomavirus. We have found that the decatenating activity present in tumor cells was 40-100 times higher than that in the rabbit liver, while no activity could be found in skin extracts. The type II topoisomerases purified from tumor and liver cells consist of two subunits with molecular masses of about 160 kDa. The conditions of the reactions of relaxation, unknotting and decatenation catalyzed by these topoisomerases II were found to be similar to those observed with enzymes of other eukaryotic cells. In the course of the purification of the VX2 enzyme, we isolated and characterized a protein of about 30 kDa in whose presence the topoisomerase II was able to catenate very efficiently supercoiled DNA molecules. This protein has the same electrophoretic mobility as an H1-2 histone, and cross-reacts with an anti-H1 antiserum. The VX2 topoisomerase II as well as the VX2 tumor should constitute useful models for assays of antitumoral drugs. PMID- 2982599 TI - Receptor-mediated endocytosis of polymeric IgA and galactosylated serum albumin in rat liver. Evidence for intracellular ligand sorting and identification of distinct endosomal compartments. AB - Rat polymeric IgA (pIgA) and galactosylated bovine serum albumin (GalBSA), once injected to rats, are avidly taken up by hepatocytes via receptor-mediated endocytosis. Of injected pIgA, 64% was transferred undigested into bile within 3 h, with a peak at 30-45 min. GalBSA was essentially digested in lysosomes. By electron microscopy using ligand-peroxidase conjugates, both ligands were internalized through coated pits/coated vesicles into similar electron-lucent vesicles and tubules. Subsequently, pIgA remained mostly associated with small vesicles clustering around and fusing with bile canaliculi, while GalBSA was predominantly found in large, heterogeneous endocytic structures and in lysosomes. By subcellular fractionation, they were associated at 3 min after injection with structures that similarly sedimented in the P fraction (250 000 - 3 X 10(6) X g X min) and equilibrated at densities of about 1.13 g/ml in sucrose gradients. At 10 min and 20 min, pIgA distribution remained mostly in the P fraction at the same equilibrium density. A minor component of the pIgA distribution was found at the density of lysosomes, but contrary to lysosomal enzymes, its distribution was not affected by Triton WR 1339. In contrast to pIgA, GalBSA was progressively recovered in the L fraction (33 000 - 250 000 X g X min) with organelles equilibrating around 1.11 g/ml, and, by 20-45 min, was found in the ML fraction (10 000 - 250 000 X g X min), around 1.20 g/ml, i.e. in lysosomes. Chloroquine did not reduce the efficiency but delayed the secretion of pIgA into bile. Similarly, it did not affect the uptake of GalBSA but apparently delayed GalBSA transfer along successive populations of host organelles. The low density, GalBSA-containing structures were devoid of proteolytic activity. Anti secretory components IgG and F(ab')2 were selectively excreted into bile, partially or totally as compounds of lower molecular mass. These antibody fragments probably result from a disulfide reduction activity along the pIgA pathway. In conclusion, our data (a) strongly suggest that pIgA and GalBSA are sorted between 3 min and 10 min after injection in non-lysosomal acidic organelles, (b) identify two successive and physically distinct endosomal populations containing GalBSA, and (c) provide the first evidence for a disulfide reduction activity along the transcytotic pathway of rat hepatocytes. PMID- 2982600 TI - Immunochemical and autoantigenic properties of the globular domain of basement membrane collagen (type IV). AB - Polyclonal rabbit antibodies raised against the globular domain NC1 of collagen IV from human placenta and a mouse tumor react with conformational antigenic determinants present on the NC1 hexamers and also with the three major subunits obtained after dissociation. The antibodies recognized unique structures within basement membranes and showed a broad tissue reactivity but only limited species cross-reactivity. Using these antibodies, it was possible to detect small amounts of collagen IV antigens from cell cultures and in serum. Monoclonal rat antibodies against mouse NC1 revealed a similar reaction potential. Autoantibodies could be produced in mice against mouse NC1 which react with kidney and lung basement membranes in a pathological manner, mimicking Goodpasture syndrome. PMID- 2982601 TI - Elution of fibronectin proteolytic fragments from a hydroxyapatite chromatography column. A simple procedure for the purification of fibronectin domains. AB - Human plasma fibronectin is composed of at least five distinct domains which we refer to as Hep-1/Fib-1, Gel, Cell, Hep-2 and Fib-2 depending on their affinity for heparin (Hep), gelatin (Gel), the cell surface (Cell) or fibrin (Fib). These domains are aligned from the NH2 to the COOH terminus in the above order and can be separated from each other by mild proteolytic digestion. We have studied the elution of fibronectin thermolysin digest from a hydroxyapatite column using a linear gradient (0.5-190 mM) of sodium phosphate buffer. The five major fibronectin domains were eluted from the hydroxyapatite chromatography column in the following order: Gel, Fib-2, Cell, Hep-1/Fib-1 and Hep-2. They were identified on the basis of their molecular mass, affinity to different macromolecules and reaction with domain-specific monoclonal antibodies. All domains except the Cell and Hep-2 domains eluted as single homogeneous peaks. The Cell domain eluted as two different peaks and the Hep-2 domain eluted as four different peaks. This is the first time that heterogeneity of these two domains has been observed. Since chromatography of a fibronectin thermolysin digest on a hydroxyapatite column provides a good separation of the five major fibronectin domains, we have elaborated a procedure in which each fibronectin domain is purified by no more than two steps; hydroxyapatite and molecular exclusion chromatography. Fractionation of fibronectin proteolytic digest on a hydroxyapatite chromatography column should be of great value in the comparative analysis of fibronectin from different sources and in the study of fibronectin heterogeneity. Its use in combination with molecular exclusion chromatography offers a simple and high-yield method for the purification of large amounts of fibronectin domains. PMID- 2982602 TI - Citrobacter lipopolysaccharides: structure elucidation of the O-specific polysaccharide from strain PCM 1487 by mass spectrometry, one-dimensional and two dimensional 1H-NMR spectroscopy and methylation analysis. AB - Structural studies on the O-specific polysaccharide of Citrobacter PCM 1487 lipopolysaccharide, using methylation analysis, Smith degradation and 1H-NMR spectroscopy, indicate that it consists of the trisaccharide repeating units (formula, see text) In this structure, 4-deoxy-D-araHex stands for 4-deoxy-D arabino-hexose. PMID- 2982603 TI - Sugar specificity and sugar-proton interaction for the hexose-proton-symport system of Chlorella. AB - The substrate specificity of the glucose-proton symport system was studied to gain information about the spatial relationship between the binding sites for glucose and proton. Charged glucose analogues such as amino sugars or sugar acids were not transported by the uptake system, with the exception of 2-amino-2-deoxy D-glucose. This glucosamine was taken up in the charged form in uniport mechanism, i.e. without symport of proton. This result was interpreted to mean that the proton-binding site of the symport system is close to the hydroxyl at carbon 2 of glucose. This interpretation was strengthened by the following facts: The steric position of hydroxyl groups at carbons 1, 2 or 3 of glucose were especially important for efficient transport. O-Methylation was not tolerated at carbon 1, but it was tolerated at carbons 3, 4 or 6. The stoichiometric flow of proton and sugar could be disturbed by removal of hydroxyl group at carbon 1 of glucose. The pH-dependence of sugar transport is sugar-specific, e.g. the amino group at carbon 2 of glucose improves transport at higher pH. The configuration at carbon 2 of glucose influences the specificity for the symported ion. It is concluded that the coupled flow of proton and glucose occurs by simultaneous coordinate movement of both in a transmembrane channel. PMID- 2982604 TI - Insulin-induced increases in the activity of the spontaneously active and ATP.Mg dependent forms of phosphatase-1 in alloxan-diabetic rat liver. AB - Liver supernatant from normal and alloxan-diabetic rats was fractionated by DEAE cellulose chromatography and the separated phosphoprotein phosphatase fractions were assayed with [32P]histone f2b, [32P]phosphorylase a and [32P]phosphorylase kinase as substrates. In diabetic rat liver, one of the phosphatase fractions found in the normal liver was significantly reduced. This fraction was identified as a mixture of the spontaneously active form and the ATP . Mg-dependent form of phosphoprotein phosphatase-1 (Fc) based on sensitivity to inhibitor-2, substrate specificity, and the fact that it could be activated 42-70% by glycogen synthase kinase-3 in the presence of ATP . Mg. Further analysis of this fraction showed that liver cytosol from diabetic rats contained 62-79% lower spontaneously active phosphatase-1 activity and 40-51% lower combined spontaneously active and ATP . Mg-dependent protein phosphatase-1 (Fc) activity. Insulin administration increased the spontaneously active and the ATP . Mg-dependent protein phosphatase 1 activities approximately 45% and 36%, respectively, in alloxan-diabetic rats. These data imply that the lower levels of spontaneously active phosphatase-1 activity in diabetic rat liver cannot be explained by presuming phosphatase-1 to have been present as Fc, the inactive form. Moreover, insulin restored the total activity of the spontaneously active and activatable forms of phosphatase-1 to those present in normal liver implying that both forms of phosphatase-1 activity are under hormonal control. PMID- 2982605 TI - Spermidine increases the accuracy of type II restriction endonucleases. Suppression of cleavage at degenerate, non-symmetrical sites. AB - The non-specific cleavage of DNA by type II restriction endonucleases (BamHI, BsuRI, EcoRI, EcoRV, HindIII, PstI and SalI) can be effectively suppressed by spermidine in millimolar concentrations, regardless of whether the non-specific cleavage is induced by high concentrations of enzyme under optimal buffer conditions or by high pH, low ionic strength, organic solvents and Mn2+ ions. The increased specificity of restriction endonucleases in the presence of spermidine is due to an enhancement of the cleavage rate at the canonical site and a slowing down of the cleavage rate at related sites. It is argued that spermidine is essential for the high accuracy of restriction endonucleases in vivo. PMID- 2982606 TI - Dioxygen-activating iron center in putidamonooxin. Electron spin resonance investigation of the nitrosylated putidamonooxin. AB - The mononuclear non-haem iron center is the dioxygen-binding site of putidamonooxin which is the dioxygen-activating component of the 4 methoxybenzoate monooxygenase. Replacement of dioxygen by nitrosyl leads to the formation of a rather stable Fe3+ X NO- complex which is characterized by electron spin resonance (ESR) at g approximately equal to 4 and g approximately equal to 2. The ESR features can be composed by two spectral components which are characterized by different tetragonal distortions of the axial symmetry. Binding of 4-hydroxybenzoate, which is the product of the enzymatic reaction, leads to the formation of an ESR spectrum with pure axial symmetry. After binding of 4 methoxybenzoate, i.e. the physiological substrate of the monooxygenase, only one spectral component, i.e. that with a small tetragonal distortion, is observed. Binding of substrate analogues, like 4-aminobenzoate and 4 trifluoromethylbenzoate, leads to a spectral heterogeneity with variable amounts of the ESR component with a large tetragonal distortion. Benzoate induces an ESR spectrum with only that spectral component with large tetragonal distortion. The iron-depleted substrate-free form of the enzyme, ligated with NO, also shows ESR heterogeneity, i.e. both spectral components overlap, with 60% of the component with large tetragonal distortion. Binding of 4-methoxybenzoate leads to the occurrence of a pure spectrum, i.e. with small tetragonal distortion, whereas binding of benzoate leads to a pure spectrum with large tetragonal distortion. Thus, the structural heterogeneity is removed by binding of both the ligand (NO) and substrate. The Fe3+ X NO- complex is discussed as an analogue of the native oxy complex Fe3+ X O2-. PMID- 2982607 TI - Mossbauer investigation of the cofactor iron of putidamonooxin. AB - Mononuclear non-heme cofactor iron of putidamonooxin has been investigated in the binary oxidized 'enzyme X substrate' complex and in the ternary 'enzyme X substrate X NO' complex via Mossbauer spectroscopy. The experimental spectra were analyzed on the basis of the spin-Hamiltonian formalism. The resulting fine and hyperfine structure parameters are compared with literature values of similar compounds. From this comparison we conclude that in the binary complex (reduced and oxidized) the mononuclear non-heme cofactor iron has a coordination number higher than four. Additionally, the cofactor iron shows remarkable spectral similarities with iron in protocatechuate 3,4-dioxygenase, though the catalytic properties of the iron sites in the two proteins are different. The data obtained form the ternary 'enzyme X substrate X NO' complex indicate that the cofactor iron (a) is in the ferric intermediate spin state (S = 3/2) and (b) is pentacoordinated, which means that upon NO binding to the reduced cofactor iron at least one ligand has to be released. Comparing our data with literature values suggests that the cofactor iron in the binary as well as in the ternary NO complex is not directly bound to a sulfur atom, though biochemical arguments seem to indicate the opposite. PMID- 2982608 TI - Influence of dA X dT and d(2aminoA) X dT base pairs on the B in equilibrium Z transition of DNA fragments. 1H-NMR study of d(C-G-C-A-m5C-G-T-G-m5C-G), d(m5C-G C-A-m5C-G-T-G-C-G) and d(C-2aminoA-C-G-T-G). AB - The Helical structures of d(C-G-C-A-m5C-G-T-G-m5C-G), d(m5C-G-C-A-m5C-G-T-G-C-G) and d(C-2aminoA-C-G-T-G) were studied in aqueous solution at various salt concentrations and temperatures by 1H-NMR spectroscopy. In 0.1 M NaCl solution only the B form was evidenced for these DNA fragments whereas in 4 M NaCl both B and Z forms, in slow exchange on the NMR time scale, were observed. Under these conditions the Z form accounted for less than 60% of the decamer conformation; conversely d(C-G)3 hexamers containing methylated cytidines were predominantly in the Z form (greater than 90%) [Tran-Dinh et al. (1984) Biochemistry 23, 1362; Cavailles et al. (1984) J. Biomol. Struct. Dyn. 1, 1347-1371]. On the other hand, d(C-2aminoA-C-G-T-G) in which the d(2aminoA) X dT base pair forms three hydrogen bonds, was found to adopt the Z conformation in 4M NaCl solution which was not the case for d(C-A-C-G-T-G) (unpublished results). The present study shows that the B in equilibrium Z transition in solution is highly sequence-dependent and that correlation exists between the stability of the duplexes (essentially governed by the number of hydrogen bonds between complementary bases) and their ability to adopt the Z conformation. PMID- 2982609 TI - Modification of the carbohydrate in ricin with metaperiodate-cyanoborohydride mixtures. Effects on toxicity and in vivo distribution. AB - Attempts to target antibody-ricin conjugates (immunotoxins) to designated cell types in vivo may be thwarted by their rapid clearance by hepatic reticuloendothelial cells which have receptors that recognise oligosaccharide side chains on the toxin. The B-chain of ricin contains high mannose type oligosaccharides and the A-chain contains a complex unit (GlcNAc)2-Fuc-Xyl-(Man)4 6, all of which potentially could be recognised by the reticuloendothelial system. Treatment of ricin with a mixture of sodium metaperiodate and sodium cyanoborohydride at pH 3.5 resulted in oxidative cleavage of the carbohydrates and reduction of the aldehyde groups thus formed to primary alcohols. By conducting the modification procedure at acidic pH, both the possibility of Schiff's base formation between the aldehyde groups and amino groups in the protein and the possibility of non-specific oxidation of amino acids were minimised. The extent of the carbohydrate modification depended on the duration of treatment, resulting maximally in the destruction of 13 of the 18 mannose residues and of all xylose and fucose. The toxicity of the modified toxin to cells in culture declined by up to 90% as the carbohydrate was destroyed. This was not due to a reduced ability of the B-chain to bind to cells or of the A chain to inactivate ribosomes. In contrast to the in vitro results, the toxicity of the modified toxin to mice and rats was elevated by up to fourfold. The modification greatly reduced the clearance of the toxin by non-parenchymal cells in the liver and prevented the damage to hepatic Kupffer and sinusoidal cells and to the red pulp of the spleen that is inflicted by the native toxin. The elevated toxicity to animals appears to be because the modified toxin evades the reticuloendothelial system and persists in the bloodstream for longer periods, thus resulting in lethal damage to vital tissues in the animal at lower dosage. The results suggest that immunotoxins prepared from modified ricin would not be readily cleared by the reticuloendothelial system and so be more effective at killing their target cells. PMID- 2982610 TI - Fructose-1,6-bisphosphatase from Synechococcus leopoliensis. Substrate-dependent dimer-tetramer interconversion. AB - Extracts of Synechococcus leopoliensis (Anacystis nidulans) contain two forms of D-fructose-1,6-bisphosphatase (EC 3.1.3.11) previously designated as forms A and B [Gerbling, K.-P., Steup, M., and Latzko, E. (1984) Arch. Microbiol. 137, 109 114]. Form B, which probably represents the major part of the total extractable fructose-1,6-bisphosphatase activity, has been purified to apparent homogeneity. Gel filtration, non-denaturing polyacrylamide gel electrophoresis, and cross linking with bis(sulfosuccinimidyl)suberate revealed that the fructose-1,6 bisphosphatase B exists in either a dimeric or in a tetrameric subform, depending upon the absence or presence of fructose-1,6-bisphosphate and Mg2+. The dimer- tetramer interconversion was readily reversible. The results provide evidence for a two-step activation of fructose-1,6-bisphosphatase B involving the reduction of the dimeric subform and the subsequent substrate-dependent conversion of the reduced dimer to a reduced tetramer, which is the only catalytically active state. In contrast to form B, no substrate-dependent interconversion was detected with form A from S. leopoliensis. PMID- 2982611 TI - ADP-ribosylation of phosphorylase kinase and block of phosphate incorporation into the enzyme. AB - Phosphorylase kinase purified from rabbit skeletal muscle was ADP-ribosylated by hen liver nuclear ADP-ribosyltransferase. This modification, as was seen in cAMP dependent phosphorylation, was observed only in alpha and beta subunits of the phosphorylase kinase and the latter was more rapidly modified. Analysis of the ADP-ribosylated amino acid residue sequenced in alpha and beta subunits showed that both subunits were modified at the area of the arginine residue. The Km for NAD was 0.10 mM and the pH optimum was 9.0. When the ADP-ribosylated phosphorylase kinase was phosphorylated by cAMP-dependent protein kinase, a reduction in phosphate incorporation occurred with increase in the ADP ribosylation. ADP-ribosylation also suppressed autophosphorylation, to a lesser degree than observed with cAMP-dependent phosphorylation. The ADP-ribosylation dependent reduction of phosphorylation resulted in a suppression of the phosphorylation-dependent activation of the phosphorylase kinase. These results together with findings of ADP-ribosyltransferase activity in the rabbit skeletal muscle [Soman, G. et al. (1984) Biochem. Biophys. Res. Commun. 120, 973-980] suggest that ADP-ribosylation participates in the regulation of the phosphorylase kinase activity through changes in the rate of phosphorylation. PMID- 2982612 TI - [Neutral glycosphingolipids of Fabry's disease lymphoblastoid lines established by Epstein-Barr virus transformation]. AB - Human lymphoid cell lines established by Epstein-Barr viral transformation of peripheral B-lymphocytes from normal subjects and from Fabry patients, were investigated for their ability to biosynthesize neutral glycosphingolipids from [14C]galactose and [14C]glucose as precursors. Galactose was taken up in the presence of high concentrations of glucose and selectively utilised by the cells in the synthesis of galactosphingolipids. The pattern of neutral glycosphingolipids labelled from [14C]galactose was slightly modified with time of labelling in either lymphoid cell line: the first labelled glycosphingolipid was lactosylceramide (LacCer) in the normal line and globotetraosylceramide (GbOse4Cer) in the Fabry line. After labelling for 96 h, a steady state was reached and the percentage of every type of labelled glycosphingolipid was stable in each cell line; however, differences in the neutral sphingolipid composition appeared between the various cell lines. When using radiolabelled glucose as precursor, the major part of the radioactivity was incorporated into neutral lipids and phospholipids; neutral sphingolipids were much less labelled than when using galactose. Catabolism of endogeneous labelled glycosphingolipids (synthesized by the cells during the 'pulse') was studied after cultivating the cells without radiolabelled precursors ('chase'). In the cells from normal subjects, all the neutral glycosphingolipids were slowly degraded (half-life time around 15-25 days for LacCer and GbOse3Cer). In contrast, in a lymphoid line from a Fabry patient, no appreciable degradation of GbOse3Cer occurred during 30 days. This block in the catabolism of GbOse3Cer is in good agreement with the previously reported deficiency of alpha-galactosidase A activity in this Fabry lymphoid cell line [Salvayre, R. et al. (1981) Biochim. Biophys. Acta 659, 445 456] and demonstrates that alpha-galactosidase B does not hydrolyze GbOse3Cer in the living cell (in contrast to the situation in vitro). PMID- 2982613 TI - Isolation and characterization of cytoplasmic poly(A) polymerase from cryptobiotic gastrulae of Artemia salina. AB - Poly(A) polymerase has been purified to near homogeneity from the cytoplasm of Artemia salina cryptobiotic gastrulae by ion-exchange chromatography on DEAE cellulose, DEAE-Sepharose CL-6B and phosphocellulose P11, gel filtration on CL Sepharose 6B, affinity chromatography on poly(A)-Sepharose 4B and ATP-agarose. The enzyme is fully dependent on exogeneous oligo(riboadenylic acid) and is free of any nuclease or other enzyme activities. In standard assay conditions the enzyme preparation has a specific activity of 5.6 mumol AMP . h-1 . (mg protein) 1. Sodium dodecyl sulphate/polyacrylamide gel electrophoresis reveals the presence of only two proteins with Mr 94 000 and 70 000. The Mr-70 000 protein has been identified as poly(A) polymerase. The enzyme is exclusively activated by Mn2+. Addition of Ca2+, Mg2+, Zn2+, NH4+, K+ or Na+ inhibits the enzymatic reaction. The activity is specific for ATP and competitive inhibition is observed in the presence of other ribonucleoside 5'-triphosphates. AMP incorporation is time-dependent and is increased non-linearly with protein and primer concentration. PMID- 2982614 TI - Further characterisation of the FAD and Fe2S2 redox centres of component C, the NADH:acceptor reductase of the soluble methane monooxygenase of Methylococcus capsulatus (Bath). AB - The absorbance contributions of the FAD and Fe2S2 redox centres of component C of the soluble methane monooxygenase complex have been resolved, using mersalyl to destroy the Fe2S2 centre. The Fe2S2 seems to be very similar to that of spinach ferredoxin, by its absorbance and electron paramagnetic resonance (EPR) spectra, and the FAD semiquinone is a neutral semiquinone. Spectrophotometry near room temperature and EPR spectroscopy near liquid-helium temperature allow the three redox couples of component C to be ordered. Component C can exist in Oe-1 (oxidised), 1e-1 (semiquinone), 2e-1 (mostly semiquinone and reduced Fe2S2), and 3e-1 forms (dihydroquinone and reduced Fe2S2), under equilibrium conditions. The ability of component C to support odd-electron forms is consistent with its proposed role as a 2e-1/1e-1 transformase, splitting electron pairs from NADH for passage to component A in one-electron steps. (The FAD appears to interact with NADH, and transfers single electrons to the Fe2S2, for donation to component A at a constant redox potential.) The mid-point potentials of component C were found using redox dyes and EPR spectroscopy and were: FAD/FAD., Em = -150 mV; Fe2S2/Fe2.S2,Em = -220 mV; FAD./FAD..,Em = -260 mV. the presence of NADH did not alter these mid-point potentials. These mid-point potentials are consistent with the role of component C as the NADH:component A reductase, passing electrons from NADH (Em = -320 mV) onto component A (Em = +150 mV and Em = -150 mV). The reducing power from NADH appears to be required by component A to activate one atom of oxygen, to insert into methane, and the reducing equivalents derived from NADH end up with the other oxygen atom, as water. PMID- 2982615 TI - cGMP-dependent protein kinase. Autophosphorylation changes the characteristics of binding site 1. AB - cGMP-dependent protein kinase binds 4 mol cGMP/mol enzyme to two different sites. Binding to site 1 (apparent Kd 17 nM) shows positive cooperativity and is inhibited by Mg . ATP, whereas binding to site 2 (apparent Kd 100-150 nM) is non cooperative and not affected by Mg . ATP. Autophosphorylation of the enzyme abolishes the cooperative binding to site 1 and the inhibitory effect of Mg . ATP. The association (K1) and dissociation (K-1) rate constant for site 2 and K1 for site 1 are not affected significantly by Mg . ATP or autophosphorylation. The dissociation rate from site 1 measured in the presence of 1 mM unlabelled cGMP is decreased threefold and over tenfold by Mg . ATP and autophosphorylation, respectively. In contrast, the dissociation rate from site 1 measured after a 500 fold dilution of the enzyme-ligand complex is 100-fold faster than that determined in the presence of 1 mM cGMP and is only slightly influenced by Mg . ATP or autophosphorylation. Only Kd values calculated with the latter K-1 values are similar to the Kd values obtained by equilibrium binding. These results suggest that autophosphorylation of cGMP-dependent protein kinase affects mainly the binding characteristics of site 1. PMID- 2982616 TI - A 1H and 31P NMR study of cis-Pt(NH3)2[d(CpGpG)-N7(2),N7(3)]. The influence of a 5'-terminal cytosine, on the structure of the cis-Pt(NH3)2[d(GpG)-N7,N7] intrastrand cross-link. AB - Proton NMR studies at 300 MHz and 500 MHz have been carried out on the trinucleoside bisphosphate d(CpGpG) and on cis-Pt(NH3)2[d(CpGpG)-N7(2),N7(3)] [abbreviated as d(CpGpGp) . cisPt]. For the Pt adduct, 13C and 31P NMR was also used for characterizing the oligonucleotide. d(CpGpG) appears to revert to a B DNA-type single helix at lower temperatures. The relatively small concentration dependence of the proton chemical shifts, in comparison with shifts due to intramolecular stacking effects, indicates that the compound is essentially single-stranded. In d(CpGpGp) . cisPt, the first nucleoside, C(1), stacks well on top of the second, G(2), despite the N conformation of the G(2) sugar ring. The platinated GpG part in this trimer adopts largely the same structure as in cis Pt(NH3)2[d(GpGpG)-N7(1),N7(2)] [den Hartog, J. H. J., et al. (1982) Nucleic Acids Res. 10, 4715-4730]. Main differences however, are changes in H8 chemical shifts and a 0.6-ppm downfield shift of the third nucleotide phosphorus, P(3), in d(CpGpGp) . cisPt with respect to P(2) in d(GpG) . cisPt. The latter shift change is likely to be induced by a structural alteration, caused by stacking of C(1) on top of G(2). Also, the large chemical shift differences between the two H8 protons in d(NpGpG) . cisPt fragments is discussed; the deviation from a mirror symmetry of the two guanine bases seems to be the main origin of this effect. The chemical shift changes, observed in the proton and phosphorus NMR chemical shift temperature and chemical shift pH profiles have been explained in terms of stack destack equilibria changes. PMID- 2982618 TI - Ovarian cancer in the young. AB - An epidemiological study of ovarian cancer in the young under 20 years old in Israel during the years 1972-1976 are presented. During these 5 years 22 new cases were reported. Germ cell tumors accounted for 63.6% and the epithelial tumor for 23%. The incidence of this group was 2.2% of the incidence of all ovarian maligancies. The five years survival was 54.5%. The less malignant tumor was dysgerminoma. There was no difference of the incidence between the various ethnic groups in this population. PMID- 2982617 TI - Identification of the Ah receptor in selected mammalian species and induction of aryl hydrocarbon hydroxylase. AB - The Ah receptor protein, important in the mechanism of induction of aryl hydrocarbon hydroxylase activity, has been identified and partially characterized in hepatic cytosolic preparations from rat, BALB/c mouse, gerbil, hamster, rabbit, ferret and guinea-pig by means of sucrose density centrifugation analysis and hydroxyapatite binding assays. Using 2,3,7,8-tetrachloro[3H]dibenzo-p-dioxin (TCDD) as the ligand, total specific binding capacities ranged over 74-691 fmol [3H]TCDD/mg cytosolic protein and apparent dissociation constants ranged over 0.30-7.8 nM. There was no quantitative correlation between the concentration of cytosolic Ah receptors and the 3-methylcholanthrene-mediated induction of aryl hydrocarbon hydroxylase activity in the species studied. Competitive binding studies with a series of monohydroxylated benzo[a]pyrene derivatives suggested the importance of electronic character in their ability to bind to the Ah receptor and to compete with TCDD for specific binding sites on the receptor. PMID- 2982619 TI - Autochthonous murine tumors: effects of viral or ultraviolet induction, immunogenicity and transplantation on intratumoral macrophages and systemic inflammatory responses. AB - Macrophage tumoricidal activity requires a constant influx of macrophages but many transplanted cancers inhibit macrophage inflammatory responses. In this paper we address the issue of whether or not autochthonous tumors induced by either mammary tumor virus (MTV) in C3H/He mice or ultraviolet (u.v.) radiation in C3H/He or BALB/c mice also depress macrophage responses. Anti-inflammatory activity was not observed either prior to or during growth of these autochthonous tumors. Rather, the opposite was observed: strongly immunogenic u.v.-induced tumors which were rejected upon transplantation to syngeneic hosts had enhanced macrophage responses and more intratumoral macrophages than those mice whose tumors were transplantable. Transplantation of MTV-induced tumors selected for more aggressive tumors which had fewer intratumoral macrophages. In both MTV- and u.v.-induced tumors inflammatory responses of mice bearing serially transplanted tumors often differed from mice with autochthonous tumors. Our results demonstrate that anti-inflammation is probably not required for emergence and growth of these autochthonous tumors, that strongly immunogenic tumors may actually enhance macrophage responses and that the effect of tumor bearing on macrophage inflammation is a characteristic of the tumor, including its site and host of origin, its immunogenicity and its transplant generation. PMID- 2982620 TI - Hormonal induction of uterine sarcomas in GR mice not dependent on mammary tumor virus genes or uterine trauma. PMID- 2982621 TI - Interstitial pneumonitis following bone marrow transplantation: pathogenesis and therapeutic considerations. AB - High-dose chemo-radiotherapy followed by allogeneic bone marrow transplantation has become standard treatment for a variety of hematological malignancies. Interstitial pneumonitis (IP) is a major complication after bone marrow transplantation, the incidence being approximately 50% (range 20-65%). Cytomegalovirus (CMV) is found in about half of these cases. If no infectious cause can be detected, the interstitial pneumonitis is labeled 'idiopathic'. The occurrence of CMV IP is related to the state of severe immunosuppression in combination with graft-vs-host disease (GvHD) in these patients. The most important factors contributing to idiopathic IP seem to be: chemotherapy, total body irradiation, agents to prevent GvHD and GvHD itself. For preventing CMV IP hyperimmune globulin seems to be the most promising method at this moment. As long as the etiology of idiopathic IP remains unclear, no measures can be taken to prevent or treat this disease. PMID- 2982622 TI - A randomised trial of cyclophosphamide pretreatment ('priming') before short duration chemotherapy for small cell lung carcinoma. AB - Forty-five patients with small cell anaplastic carcinoma of the bronchus were treated with four four-weekly courses of a combination of cyclophosphamide, vincristine and methotrexate. Randomisation was carried out to determine whether they received in addition 1 g/m2 of cyclophosphamide 1 week before the three-drug therapy. Patients with limited disease received radiotherapy after their chemotherapy. Myelosuppression was similar in the two groups, but the additional cyclophosphamide did not improve remission duration or survival. Confining the chemotherapy to four courses did not give shorter survival times to those reported in other studies. PMID- 2982623 TI - Organization of cytoskeleton and fibronectin matrix in Rous sarcoma virus (RSV) transformed fibroblast lines with different metastatic potential. AB - Metastatic clones growing in 0.6% 'hard' agar were selected from the non metastatic Rous sarcoma virus (RSV)-transformed tumorigenic B77-3T3 mouse fibroblast line. The incidence of spontaneous lung metastases varied among clones around 100%, while it was lower than 5% in the parental tumor line. The organization of microfilaments, microtubules and intermediate filaments as well as the pattern of extracellular fibronectin matrix were analyzed by immunofluorescence in two representative clones (B77-AA6 and B77-AA12) and was compared with the structural features displayed by a highly metastasizing RSV induced mouse sarcoma line (SR-BALB). In the metastatic clones studied microtubules and intermediate filaments were similarly organized in a pattern not significantly different from that of the non-metastatic parental cell line. The major finding was a marked concentration of actin-containing structures in the periphery of cells and notably at the level of surface protrusions, suggesting a high surface motility. In the same lines the production of fibronectin and its distribution in the cell layer and culture medium were analyzed. Metabolic labelling and immunofluorescence experiments indicated that the nonmetastasizing cells (B77-3T3) retain higher amounts of fibronectin in the cell layer and organize this molecule in extracellular fibers, while the metastatic clones (B77 AA6 and B77-AA12) as well as the metastatic line (SR-BALB) are unable to retain and organize fibronectin at their surface. This paper shows that the progression of tumorigenic cell lines toward a metastatic phenotype involves a redistribution of cytoskeletal actin and a loss of organized fibronectin matrix. PMID- 2982624 TI - Characterization of tumor cell lines from a spontaneous rat sarcoma expressing an endogenous retrovirus. AB - We have characterized various biologic, immunologic and growth properties of several cell lines established from a spontaneous rat sarcoma that was discovered more than 60 yr ago. The tumors consisted of mixed cell types with no detectable host cellular immune response. Cultures derived from single-cell clones of the parental cell line were non-invasive but highly tumorigenic even in adult rats. The cultured cells spontaneously released replication-competent endogenous rat type C virus which did not carry a transforming gene in its genome. Since normal cells from the same rat strain did not produce a retrovirus, it is possible that production of the endogenous retrovirus may have triggered specific cellular changes necessary for the oncogene expression and development of this tumor. PMID- 2982625 TI - Recent progress in studies of polyomavirus tumour antigens. AB - The polyomavirus tumour (T) antigens were originally identified by their reactivity with antisera from tumour-bearing animals. The primary structure of the three T-antigens has been established by combining the information from the nucleotide sequencing of DNA, RNA analysis, and peptide mapping. The functions of the T-antigens in productive infection and cellular transformation have largely been analysed by using virus mutants. The large T-antigen binds specifically to polyomavirus DNA. This binding is probably linked to the activity of the protein in the control of viral DNA and RNA synthesis. In addition, the large T-antigen has the ability to confer an unlimited growth potential to cells in culture. The middle T-antigen is a primary inducer of cellular transformation. The part of this protein that is located in the plasma membrane, is associated with a tyrosine kinase activity. The small T-antigen, finally, has not yet been studied extensively. However, small T-antigen has to be expressed to allow a complete productive infection cycle in mouse cells. PMID- 2982627 TI - Prolyl hydroxylase production can be uncoupled from the regulation of procollagen synthesis. AB - Primary avian tendon (PAT) cells increase the production of procollagen from 10 12% to 40-50% of total protein synthesis in response to the addition of ascorbate and an increasing cell density. We now show that prolyl hydroxylase (PH) also increases its activity by greater than five-fold in response to increasing cell density; but unlike procollagen production, this is independent of the presence of ascorbate. The increased activity is a result of greater enzyme production and not a shift in the ratio of inactive to active forms which remains constant at about 10% of the total enzyme proteins. We present the possibility that at low cell density the levels of PH activity could limit production of collagen. PMID- 2982626 TI - Differential structural requirements for the induction of cell attachment, proliferation and differentiation by the extracellular matrix. AB - The subendothelial extracellular matrix (ECM) mediates the attachment of both human Ewing's sarcoma and colon carcinoma cells. Attachment and flattening of the sarcoma cells was sensitive to heat treatment but not to periodate oxidation of the ECM, whereas the colon carcinoma cells attached and flattened over heated but not periodate-treated ECM. Such differential sensitivity to heat treatment and periodate oxidation was also observed using purified fibronectin and laminin, respectively, but the inhibition of cell attachment was greater than with a similarly treated ECM. It is therefore conceivable that fibronectin and laminin specifically mediate the attachment and flattening of Ewing's sarcoma and colon carcinoma cells to the ECM, but that other constituents may support this attachment either directly or via interaction and stabilization of adhesive glycoproteins in the ECM. The ECM-mediated morphological differentiation of adult rat oligodendrocytes was sensitive to periodate oxidation to a much higher extent than to heat treatment of the ECM. In contrast, both treatments had only a small effect on the ECM-induced proliferation of vascular endothelial cells. These results indicate that different constituents of the ECM may be held responsible for its effects on different parameters of cell behavior, and that various cell types respond differently to a given modification of the ECM. PMID- 2982629 TI - Mitogenesis induced by calcium-containing crystals. Role of intracellular dissolution. AB - Hydroxyapatite, like other calcium-containing crystals previously studied by us, is mitogenic for cultured human fibroblasts. This mitogenic effect is not a result of increased ambient calcium concentration due to extracellular crystal dissolution. Synthetic crystals labelled uniformly with calcium 45 (45Ca) undergo endocytosis when incubated with cells and are solubilized. Such solubilization is inhibited by chloroquine or ammonium chloride in concentrations that significantly block the mitogenic effect of crystals but not that of serum. The data suggest that mitogenesis and intracellular crystal dissolution are related phenomena. PMID- 2982628 TI - Similar sensitivity of human lymphoblast cells proficient or deficient in nucleotide biosynthesis to the cytotoxic and cell cycle effects of 3 aminobenzamide. AB - 3-Aminobenzamide (3AB), a potent inhibitor of poly(ADP-ribose) polymerase, induces cytotoxicity and cell cycle delays in exposed cells. 3AB has been reported to inhibit de novo nucleotide synthesis in human lymphoblasts. To determine if the 3AB-mediated effects are related to this inhibition of DNA synthesis, cytotoxicity and cell cycle progression in human lymphoblastoid cell lines, proficient or deficient in salvage nucleotide synthesis pathways, were determined after exposures to 3AB. In addition, changes in cell cycle progression were followed after treatment with 3-aminobenzoic acid (ABA), a nonactive analog of 3AB, and 5-methylnicotinamide (5MN), a less potent inhibitor of poly(ADP ribose) synthesis. All three lines responded similarly to the different treatments. Cells deficient in salvage nucleotide synthesis pathways were no more sensitive to the cytotoxic or cell cycle effects of 3AB, ABA or 5MN. Thus, effects on nucleotide synthesis are not responsible for either cytotoxicity or the retarded cell growth found in human lymphoblastoid cells exposed to inhibitors of poly(ADP-ribose) synthesis. PMID- 2982630 TI - Chediak-Higashi syndrome: studies in long-term bone marrow culture. AB - The granulocyte defects of Chediak-Higashi syndrome (CHS) include neutropenia, characteristic giant lysosomal granule morphology, and functionally abnormal cell motility, degranulation, and bacterial killing. Findings of elevated levels of adenosine 3', 5' cyclic monophosphate nucleotide (cAMP) and of concanavalin A (Con A) capping have suggested a pathogenic role of a cyclic-nucleotide-related defect in microtubule polymerization, but not all patients exhibit these abnormalities. In order to test which defects derive from the cells' genetic program and which from the host environment, we examined granulocytes produced by CHS bone marrow progenitors in long-term in vitro bone marrow cultures. These cells exhibited the characteristic giant-granule morphology and defective cell motility of CHS. However, culture-derived CHS granulocytes had normal cAMP contents and normal spontaneous capping of Con A. Granulopoiesis diminished dramatically after five weeks in culture, with accompanying autophagocytosis by mononuclear phagocytes. In mixing experiments, the phenotype of the mature granulocytes corresponded to the genotype of the hematopoietic component of the culture rather than the stroma. These results indicate that the hallmark giant granule morphology and cell motility defect of CHS are expressions of the genetic program of the hematopoietic cells. However the abnormalities in resting cyclic nucleotide levels and in Con-A capping may be secondary manifestations of the disease and are not essential to the pathogenesis of the chemotactic defect. PMID- 2982631 TI - Sedimentation rate, leucocytes, platelet count and haemoglobin in bronchial carcinoma: an epidemiological study. AB - Erythrocyte sedimentation rate (ESR), leucocyte count, platelet count, haemoglobin concentration (Hb) and survival were studied in an epidemiological material of 258 patients with bronchial carcinoma. The mean ESR was elevated (48.1 mm/h) and, more so in squamous cell and small cell carcinoma than in large cell and adenocarcinoma. Leucocytosis, over 9 X 10(9)/l, was found in 39% of the patients and thrombocytosis, over 400 X 10(9)/l, in 13% of the patients. Nine per cent of the patients were anaemic, with Hb lower than 11 g/l. There was a statistically significant positive correlation between ESR, leucocyte count and platelet count, but a negative correlation was found between these 3 variables and Hb. ESR, leucocyte count and platelet count also correlated to survival at a statistically significant level. Multiple regression analysis of these laboratory variables showed that they explain about 8% of the great variations in survival time. PMID- 2982632 TI - Assessment of percussion, vibratory-shaking and breathing exercises in chest physiotherapy. AB - While gravity-assisted positions (postural drainage) and the forced expiratory technique are known to promote sputum clearance, the additional value of percussion, vibratory-shaking and breathing exercises individually in chest physiotherapy is uncertain. These modalities have been evaluated in 8 patients with copious sputum production (mean: 44 g/day), using an inhaled radioaerosol technique. Tracheobronchial clearance was unaffected by the addition of either vibratory-shaking or percussion with and without breathing exercises to postural drainage. There was however a significant (p less than 0.01) increase in the dry weight of sputum produced during each of these treatments. The combination of postural drainage used in conjunction with the forced expiration technique is responsible for the majority of mucus mobilisation and should form the basis of routine chest physiotherapy programmes; the other modes appear to be of lesser value. PMID- 2982633 TI - Role of the cat substantia nigra pars reticulata in eye and head movements. II. Effects of local pharmacological injections. AB - Guided and reflex eye movements were studied in cats trained to make orienting saccades toward visual and auditory targets. Injections of a GABA-agonist (Muscimol) or GABA-antagonists (Bicuculline and Picrotoxin) were made in the Substantia Nigra pars reticulata (SNpr). Bicuculline and Picrotoxin, whether unilaterally or bilaterally injected had no effect on the posture nor the oculomotor performance of the animals. Neck muscle activity remained symmetrical. Unilateral injections of Muscimol produced oro-facial akinesia, reduction of the number of eye movements, contralateral head turning, visual neglect mostly (but not only) for ipsilateral visual space. Balance between the gains of the vestibulo-ocular reflex (VOR) in the two directions of movement was changed. Gain was decreased for the ipsilateral rotation. The optokinetic nystagmus (OKN) was not affected. Contralateral neck muscles were hypertonic. After bilateral injections of Muscimol, the cats did not orient. The VOR was normal when the injections induced no postural asymmetry. Hypertony was bilateral. Implications of these results for the role of the basal ganglia in motor control are discussed. We suggest that in Parkinson's disease the fixed inhibitory drive of the SNpr on the tectum and on the thalamus is disrupted. PMID- 2982634 TI - Effects of medial midbrain lesions on thermoresponsive neurons in the thalamus of the rat. AB - This study is concerned with midbrain influences on the transmission of thermal information from the rat's scrotal skin to the specific thalamus. Single unit recordings were made from neurons in the ventrobasal thalamus which responded to scrotal warming. After a thermoresponsive neuron had been identified, a small area of the medial midbrain was electrolytically lesioned and the thermal response of the unit was tested again. Lesioning the midbrain raphe nuclei, centralis or dorsalis, or parts of the central grey matter was always followed by a complete loss of thermal responsivity. Lesions which destroyed the rostral end of the nucleus raphe centralis together with parts of the adjacent reticular formation had no effect on thalamic discharge rates. It is concluded that the midbrain raphe nuclei and the central grey matter form an essential part of the extralemniscal pathways which transmit peripheral thermal information to the rat's thalamus. PMID- 2982635 TI - Hippocampal slices of kindled rats reveal calcium involvement in epileptogenesis. AB - Daily repeated tetanic electrical stimulation (kindling) of the brain may cause a long term enhancement of synaptic transmission and epileptiform activity of progressive severity and generalisation, eventually leading to spontaneous seizures. Evidence for a cellular mechanism underlying kindling has been obtained in vitro in slices from the hippocampus of kindled rats. A marked enhancement in extracellular calcium changes, induced by electrical stimulation or by iontophoresis of excitatory aminoacids was found in kindled tissue. This implies that changes in dendritic calcium conductances are involved in kindling epileptogenesis. PMID- 2982636 TI - Topographic organization of fimbria-fornix fibers projecting to the lateral septum of rats: a single and field response analysis. AB - Single-unit and field potential responses, evoked by stimulation of restricted fimbria-fornix (fi-fx) segments, were recorded over the entire extent of the lateral septal complex (LSC) of rats in order to examine electrophysiologically the topographic organization of fi-fx fibers projecting to the LSC. We confirmed earlier studies showing that fi-fx fibers constitute a bilateral, monosynaptic excitatory projection to the LSC. The projection appeared to be topographically ordered in that medial fi-fx fibers innervated neurons in the mediodorsal part of the LSC whereas fibers coursing through progressively more lateral positions in the fi-fx activated neurons in gradually more lateral and ventral parts of the LSC. In addition to the topographic organization, a marked convergence of the fi fx fibers on neurons located in the dorsolateral part of the LSC was apparent from both the single-unit and field potential experiments. PMID- 2982637 TI - Leishmania mexicana: purine-metabolizing enzymes of amastigotes and promastigotes. AB - Cultured promastigote and isolated amastigote forms of Leishmania mexicana mexicana have been surveyed for the presence of enzymes involved in purine metabolism. Quantitative but not qualitative differences between the enzymes of two forms were discovered. There were found to be significant differences between the enzyme content of L. m. mexicana and that reported for L. donovani. Extracts of both parasite forms of L. m. mexicana were found to have higher levels of adenine deaminase (EC 3.5.4.2) and guanine deaminase (EC 3.5.4.3) than adenosine deaminase (EC 3.5.4.4). There appeared to be two distinct nucleosidases (EC 3.2.2.1), one active on nucleosides, the other on deoxynucleosides. Phosphorylase (EC 2.4.2.1) could be detected only in the catabolic direction. Nucleotidases were present, but were more active on 3' (EC 3.1.3.6)- than 5' (EC 3.1.3.5) nucleotides. Phosphoribosyltransferase (EC 2.4.2.7,.8 and .22) and nucleoside kinase (EC 2.7.1.20) activities were detected in both forms. Nucleotide interconverting enzymes were found to be present, with IMP dehydrogenase (EC 1.2.1.14) being the most active. Cell fractionation experiments revealed that, in the promastigote, enzyme separation within the parasite may play an important part in regulating cellular purine metabolism. PMID- 2982638 TI - Leishmania mexicana: enzyme activities of amastigotes and promastigotes and their inhibition by antimonials and arsenicals. AB - A major difference between the metabolism of Leishmania species amastigotes and cultured promastigotes was found in the area of CO2 fixation and phosphoenolpyruvate metabolism. Malate dehydrogenase (EC 1.1.1.37) and phosphoenolpyruvate carboxykinase (EC 4.1.1.49) were at much higher activities in amastigotes than promastigotes of both L. m. mexicana and L. donovani, whereas the reverse was true of pyruvate kinase (EC 2.7.1.40). Pyruvate carboxylase (EC 6.4.1.1) and malic enzyme (carboxylating) (EC 1.1.1.40) could not be detected in L. m. mexicana amastigotes. Promastigotes of L. m. mexicana had a high NAD-linked glutamate dehydrogenase activity in comparison to amastigotes, whereas NADP linked glutamate dehydrogenase activity was detected only in amastigotes. Leishmania m. mexicana culture promastigotes were killed in vitro by the trivalent antimonial Triostam (LD50, 20 micrograms/ml) and the trivalent arsenical melarsen oxide (LD50, 20 micrograms/ml), but they were unaffected by Pentostam. Neither antimonial drug significantly inhibited leishmanial hexokinase (EC 2.7.1.2), phosphofructokinase (EC 2.7.1.11), pyruvate kinase, malate dehydrogenase or phosphoenolpyruvate carboxykinase, whereas melarsen oxide was a potent inhibitor of all the enzymes tested except phosphoenolpyruvate carboxykinase. PMID- 2982639 TI - Ascaris lumbricoides: characterization of the collagenous components of the adult cuticle. AB - The proteins of the cuticle of adult Ascaris lumbricoides suum were characterized with respect to heterogeneity, glycosylation, and susceptibility to collagenase. Pepsin digestion of intact cuticles was used to determine the extent of stable triple-helical structures of the cuticular components. With sodium dodecyl sulfate-poly-acrylamide gel electrophoresis, it was shown that treatment of purified cuticles with beta-mercaptoethanol released three components (99, 90, and 68 kDa) which comprise 95% of the total solubilized material. The remaining fraction consists of at least four components (16, 28, 154, and 173 kDa). Periodic acid-Schiff staining showed that the only glycoprotein was the 173-kDa component. All cuticular components, except the 173-kDa protein, were degraded by bacterial collagenase. Pepsin digestion of intact cuticles for 24 hr at 4 C produced, after reduction, a 95-kDa fragment; by 96 hr, four fragments (95, 90, 83, and 77 kDa) were evident. When the 96-hr pepsin digest was treated with fresh pepsin, the 77-kDa fragment became the major constituent. With agarose gel electrophoresis, analysis of non-reduced, pepsin-released material revealed intact aggregates that were greater than 2 X 10(3) kDa. The enzyme digestion studies indicate that, with the exception of the 173-kDa component, each cuticular protein contains collagenous domains and that, within the cuticle, the longest contiguous collagen chain in a triple-helical conformation has a uniform molecular size of 77 kDa. PMID- 2982640 TI - Influence of hydrochlorothiazide on the pain threshold and on the antinociceptive activity of morphine, in rats. AB - Hydrochlorothiazide, acutely injected in rats, has a weak analgesic activity per se and potentiates and prolongs the antinociceptive effect of morphine. PMID- 2982641 TI - Hormonal induction of steroid sulphatase in the mouse. AB - By comparing steroid sulphatase levels per se, and also ratios to alpha galactosidase, in 6 sets of mice - normal females, entire and castrated males both with and without exogenous testosterone administration - we obtained support for the contention that induction of this enzyme is in part controlled by male hormones. PMID- 2982642 TI - Methylation of hippocampal phosphatidylethanolamine and proteins during long lasting potentiation. AB - Whereas the monomethylation of hippocampal phosphatidylethanolamine is decreased following the induction of long-lasting potentiation of the CA1 population spike, carboxymethylation of proteins is unaffected. PMID- 2982643 TI - Cleavage of des-Arg9-bradykinin by angiotensin I-converting enzyme from pig kidney cortex. AB - Fast and very slow hydrolyses of des-Arg9-bradykinin and angiotensin II by angiotensin I-converting enzyme were detected by high performance liquid chromatography. The Michaelis constants of the enzyme, Km values, for des-Arg9 bradykinin and bradykinin were found to be 0.24 mM and 4.4 microM, and the maximum velocities, Vmax values (mumol . min-1 . mg protein-1) for these compounds to be 3.24 and 0.34, respectively. The enzyme also hydrolyzed Z-Gly-Pro Gly-Gly-Pro-Ala to a tripeptide that was identified as dansyl-Gly-Pro-Ala by TLC on polyamide. These observations show that the enzyme hydrolyzes the peptides at the bond before the prolyl residue in the penultimate position. PMID- 2982644 TI - Effect of mycotoxin (T-2 toxin) on catecholamine and Na+, K+-ATPase levels in rat epididymis. AB - The effect of mycotoxin (T-2 toxin) on catecholamines and Na+, K+-ATPase activities in rat epididymis has been evaluated. Dopamine and norepinephrine levels were significantly elevated in the caput and corpus regions whereas their levels remained unchanged in the caudal part of the epididymis. Na+, K+-ATPase activity was significantly increased in all the three regions of rat epididymis as a result of the toxin treatment. These changes may suggest an adverse effect on epididymal functions in rats. PMID- 2982645 TI - Reduced collagen biosynthesis in oral mucosa of beige (Chediak-Higashi) mice. AB - Oral mucosa of beige (Chediak-Higashi) mice had decreased levels of collagen synthesis and prolyl hydroxylase activity compared with normal animals. No significant difference was observed in non-collagen protein synthesis between the two groups. These results suggest that decreased collagen biosynthesis in oral tissues may be partially involved in the increased incidence of periodontal disease in the Chediak-Higashi syndrome. PMID- 2982647 TI - Enhanced expression of Epstein-Barr virus early antigens by antitubulin agents in a latently infected human lymphoblastoid cell line. AB - 28 anticancer agents have been surveyed for Epstein-Barr virus (EBV) activating potency. Two vinca alkaloids with antitubulin activity, vinblastine (VLB) and vincristine (VCR), enhanced the expression of EBV early antigens (EA) in a latently infected human lymphoblastoid cell line (Raji), when used in combination with n-butyrate. Other antitubulin agents, colchicine, colcemid, and podophyllotoxin, had the same effect, although their effects were less than that of the potent tumor promoter, 12-O-tetradecanoyl phorbol-13-acetate (TPA). PMID- 2982646 TI - Adenosine uptake in pyrimidine 5'-nucleotidase deficient human erythrocytes via a high affinity transport system. AB - Using a pulse-labeling technique, 14C-adenosine uptake into pyrimidine 5' nucleotidase (P5N) deficient erythrocytes (RBC) was found to be impaired. The Lineweaver-Burk plot showed Km values of 2.0 X 10(-3) mM and 0.2 X 10(-3) mM for normal RBC and P5N deficient RBC, respectively. These results indicate that P5N is one of regulators of the adenosine transport system and/or is associated with adenosine carrier protein. PMID- 2982649 TI - Membrane resting potentials in cultured mouse neuroblastoma cells. AB - Membrane resting potentials (MRP) were measured systematically in cultured mouse N2A neuroblastoma cells: in the logarithmic growth phase; in subconfluent cultures; in confluent cultures; after dBcAMP had induced morphological differentiation. Neurite extension was accompanied by a significant increase in MRP as compared to the appropriate controls. No significant differences in MRP were observed with regard to the different growth phases. PMID- 2982648 TI - The use of indentometry to study the effect of agents known to increase skin c AMP content. AB - Local, externally applied pharmacological agents which are assumed to raise the c AMP level, decrease the low pressure indentation value of the forehead skin of certain human volunteers. PMID- 2982650 TI - Effects of retinoic acid on ascites cells of the TA3 mouse mammary carcinoma. AB - Retinoic acid caused a decrease in adhesiveness but no growth change in the allotransplantable TA3-Ha cell and no change in adhesiveness or growth in the strain specific TA3-St cell. The retinoic acid binding protein was detected in the TA3-Ha, but not the TA3-St, cell. PMID- 2982651 TI - Variability in the nucleic acid binding site size and the amount of single stranded DNA-binding protein in Escherichia coli. AB - The Escherichia coli single-stranded DNA binding protein (SSB), essential for DNA replication, recombination and repair, can undergo a thermally induced irreversible conformational change which does not eliminate its biological activity, but changes the number of nucleotides it covers (binding site size) when binding to a single-stranded nucleic acid lattice. The binding site size of native and conformationally changed SSB was also found to be a function of the molecular mass of the polynucleotide, an observation which is unusual for single stranded DNA binding proteins and will greatly affect the affinity relationship of this protein for nucleic acids. A radioimmunoassay used to quantitate in SSB level in cells revealed the number of SSB tetramers to be larger than initial estimates by a factor of as much as six. All these data suggest that the biological role of SSB and its mechanism of action is by far more complex than originally assumed. PMID- 2982652 TI - Light-induced conformational change in rhodopsin detected by modification of G protein binding, GTP gamma S binding and cGMP phosphodiesterase activation. AB - CNBr treatment of rod outer segments was performed in dark and in light conditions. With the subsequent modified rhodopsin and opsin the cGMP phosphodiesterase activation system was reconstituted. The recombination systems exhibited greatly reduced G-protein binding, GTP gamma S binding and cGMP phosphodiesterase activation. The reduction in activity of these three steps of the PDE activation cascade is most significant with modified opsin and is shown to be due to its inability to bind the G alpha subunit. The correlation between the localization of CNBr cleavage in dark and light conditions and these results is strongly indicative that a light-induced conformational change occurs in two extradiscal regions of rhodopsin. PMID- 2982653 TI - Maximal activities of enzymes involved in adenosine metabolism in muscle and adipose tissue of rats under conditions of variations in insulin sensitivity. AB - The maximal activities of 5'-nucleotidase, adenosine deaminase and adenosine kinase were measured in quadriceps or soleus muscle from animals in which the sensitivity to insulin was changed. Most conditions caused no effect on the activities but exercise-training increased the activity of adenosine deaminase and cold exposure increased the activity of 5'-nucleotidase in soleus muscle: in addition, ageing decreased markedly the activities of all three enzymes in both muscles. When the activities are based on mg protein they are much higher in both white and brown adipose tissue than in muscle, suggesting that changes in adenosine concentration may be important in changing insulin sensitivity in adipose tissue whereas changes in adenosine receptor number may be more important in muscle. PMID- 2982654 TI - Salt-stable association of simian virus 40 capsid with simian virus 40 DNA. AB - In 8 M CsCl, a fraction of the wild-type previrions and tsB228 nucleoprotein complexes lose their core histones but retain their capsid. These histone depleted complexes appear in the electron microscope as a protein shell attached to supercoiled DNA. Consistent with this result, we find that in 1 M NaCl, the wild-type previrions dissociate into two populations of nucleoprotein complexes. One population sediments between 50 and 140 S and morphologically resembles the shell-DNA complexes isolated in CsCl gradients. The other population is comprised primarily of nucleoproteins which sediment at 40 S. PMID- 2982655 TI - On the 6-phosphofructo-1-kinase phosphatase activity of protein phosphatase 2C and its dimeric nature. AB - A recently described 6-phosphofructo-1-kinase phosphatase (PFK-phosphatase) shared several properties with protein phosphatase 2C, but exhibited differences with respect to molecular mass and substrate specificity. Chromatography on histone-Sepharose, gel filtration experiments on Sephacryl S-200 and Sephadex G 100 as well as sucrose density gradient centrifugation show that both enzyme preparations behave identically under all experimental conditions used. The low activity of PFK-phosphatase phosphorylated histone H2B had resulted from an inhibition of the enzyme by high concentrations of this substrate. The apparent molecular mass of protein phosphatase 2C as calculated from Sephacryl chromatography and sedimentation analysis is about 90 kDa, the molecular mass obtained by SDS gel electrophoresis about 45 kDa. The native enzyme therefore seems to be a dimer consisting probably of 2 identical subunits. Accordingly, the previously described PFK-phosphatase is protein phosphatase 2C. PMID- 2982656 TI - Effects of bc1-site electron transfer inhibitors on the absorption spectra of mitochondrial cytochromes b. AB - Changes are described that are brought about by antimycin, NoHOQnO, funiculosin, myxothiazol and mucidin in the alpha-, beta- and gamma-absorption bands of reduced and oxidized cytochromes b in the isolated complex bc1 form beef heart mitochondria. The inhibitors can be divided into 2 groups. Antimycin, funiculosin and NoHOQnO are likely to shift the spectrum of b-562 and compete for specific binding with complex bc1, with each other but not with myxothiazol and mucidin. The spectral effects of the latter two inhibitors are more difficult to interpret and may involve contributions not only from b-562 but from b-566 as well. The existence of 2 independent inhibitor binding-sites in the complex bc1 corroborates the Q-cycle hypothesis. PMID- 2982658 TI - Vasopressin inhibits cyclic AMP accumulation and adenylate cyclase activity in cerebral preparations. AB - Arginine-vasopressin (AVP) and lysine vasopressin (LVP) elicited a dose-dependent inhibition of noradrenaline-sensitive cyclic AMP accumulation in rat cerebral cortical slices, and of forskolin-stimulated adenylate cyclase activity in a rat cerebral cortical membrane preparation. In both cases LVP was more potent than AVP, and exerted half-maximal effects at concentrations similar to those found effective in binding studies on rat hippocampal membranes. In hippocampal slices, AVP did not affect cyclic AMP accumulation at low concentrations but potentiated the effect of noradrenaline at higher concentrations. In caudate membranes, AVP inhibited dopamine-stimulated adenylate cyclase with a similar dose-dependency to that seen for forskolin activation in cortex membranes. PMID- 2982657 TI - Inhibitory effect of atrial natriuretic factor on adenylate cyclase activity in adrenal cortical membranes. AB - The effect of rat synthetic atrial natriuretic factor (ANF) on adenylate cyclase activity was studied in adrenal cortical membranes. Synthetic ANF (Arg 101-Tyr 126) inhibited adenylate cyclase activity in a concentration-dependent manner. The maximum inhibition observed was about 25% with an apparent ki of 5 X 10(-11) to 10(-10) M. Various hormones such as isoproterenol, dopamine, prostaglandin (PGE1) and ACTH-stimulated adenylate cyclase to various degrees and ANF inhibited the stimulatory effect of all these hormones to some degree but never abolished it. In addition, ANF was also able to inhibit the stimulation exerted by forskolin which activates adenylate cyclase by receptor-independent mechanism. This is the first study demonstrating the inhibitory effect of ANF on adrenal cortical adenylate cyclase. From these results it can be suggested that the inhibition of adenylate cyclase may be one of the mechanisms through which ANF exerts the inhibitory effect on steroidogenesis stimulated by various hormones and agents. PMID- 2982659 TI - Relationships between phosphoinositide metabolism, Ca2+ changes and respiratory burst in formyl-methionyl-leucyl-phenylalanine-stimulated human neutrophils. The breakdown of phosphoinositides is not involved in the rise of cytosolic free Ca2+. AB - The relationships between the changes of cellular Ca2+, the activation of phosphoinositide turnover and the functional responses induced by the stimulus receptor interactions in neutrophils are matter of controversy. By measuring the concentration dependency of different formyl-leucyl-methionyl-phenylalanine (FMLP)-induced changes, the following values of ED50 were found: 1.6 and 0.8 nM for the rise in [Ca2+]i monitored with Quin-2, in the presence and absence of exogenous Ca2+, respectively; 20 nM for the activation of phosphoinositide metabolism, monitored as change in the 32Pi of phosphatidate; 14 nM for membrane bound Ca2+ mobilization, monitored with chlorotetracycline (CTC); 34 nM for 45Ca2+ influx and 32 nM for the respiratory burst. Furthermore, low dose of FMLP causes an increase in [Ca2+]i in absence of activation of breakdown of phosphatidylinositol, phosphatidylinositol 4-monophosphate and phosphatidylinositol 4,5-biphosphate monitored as changes in [3H]glycerol radioactivity. The results clearly demonstrate that the increase in [Ca2+]i, due to the release from intracellular stores, is not caused by the breakdown of phosphatidylinositides. On the other hand, the data of the similarity of ED50 are compatible with an involvement of phosphoinositide response in the release of membrane bound Ca2+, monitored with CTC, and in the 45Ca influx and in the respiratory burst. PMID- 2982661 TI - The effect of rhodopsin phosphorylation on the light-dependent activation of phosphodiesterase from bovine rod outer segments. AB - ATP quenches light-dependent phosphodiesterase (PDE) activation in rod outer segments presumably due to rhodopsin phosphorylation. Here we compared the efficiency of phosphorylated and non-phosphorylated rhodopsins as PDE activators in a reconstituted cell-free system. It is shown that the ability of phosphorylated membranes to activate this enzyme is suppressed compared with non phosphorylated ones. PMID- 2982660 TI - Breakdown and synthesis of polyphosphoinositides in fMetLeuPhe-stimulated neutrophils. AB - The interconversions of the inositol-containing lipids (PI, PI-P and PI-P2) and their products (DG, inositol phosphates and PA) in human and rabbit neutrophils stimulated with fMetLeuPhe and PMA have been examined. PMA causes only the phosphorylation of PI to PI-P whereas fMetLeuPhe causes phosphorylation of both PI and PI-P yielding PI-P2 and the hydrolysis of all three lipids. While the predominant reaction is breakdown of PI to PA catalysed by phospholipase D, approx. 2% of PI is converted to polyphosphoinositides and then broken down by the phospholipase C route yielding inositol phosphates and DG. The latter reaction occurs without detectable lag and is a function of receptor occupancy. The amount of inositol trisphosphate thus formed would be sufficient to release Ca2+ from intracellular stores. PMID- 2982662 TI - A specific acylating agent for the [3H]phencyclidine receptors in rat brain. AB - A derivative of phencyclidine (PCP, 1 in fig. 1) bearing an isothiocyanate moiety on the meta position of the aromatic ring (Metaphit, 3 in fig. 1) has been synthesized and identified as a rapid and specific site-directed acylating agent of the [3H]phencyclidine binding site in rat brain homogenates. The percentage of sites irreversibly inactivated by Metaphit was found to be the same in the hippocampus and striatum and the remaining sites were unaffected by Metaphit treatment under any conditions, suggesting that at least two distinct binding sites are present. An isomeric isothiocyanate derivative did not irreversibly inhibit [3H]phencyclidine receptors, indicating structural specificity for Metaphit in the inhibition of these receptors. The availability of Metaphit should greatly facilitate study of the structure and function of the phencyclidine receptors. PMID- 2982663 TI - Partial uncoupling, or inhibition of electron transport rate, have equivalent effects on the relationship between the rate of ATP synthesis and proton-motive force in submitochondrial particles. AB - The rates of electron transport and of ATP synthesis have been measured in bovine heart Mg-ATP submitochondrial particles oxidising succinate under conditions of partial attenuation of the proton-motive force by malonate or FCCP. This paper reports evidence that the relationship between the rate of ATP synthesis and the magnitude of the proton motive force is independent of the mode by which the decrease of the proton motive force is achieved. PMID- 2982664 TI - Prostaglandins E1 and E2 enhance the stimulation of superoxide release by 1 oleoyl-2-acetylglycerol from human neutrophils. AB - Superoxide release from human neutrophils was stimulated either by receptor activation (using fMet-Leu-Phe) or by activating, independently, each of the two pathways considered to be involved in signal transduction--calcium mobilization (using the ionophore, A23187) and protein kinase C activation (using phorbol myristate acetate or 1-oleoyl-2-acetylglycerol). Prostaglandin E1 (3 X 10(-5) M) decreased fMet-Leu-Phe-stimulated superoxide release, had no effect on superoxide release stimulated by A23187, or by phorbol myristate acetate, and markedly enhanced the superoxide release stimulated by 1-oleoyl-2-acetylglycerol. Similar enhancement was obtained with prostaglandin E2. PMID- 2982665 TI - Isolation of the aspartokinase domain of bifunctional aspartokinase I-homoserine dehydrogenase I from E.coli K12. AB - A proteolytic fragment (Mr approximately 25 000) carrying only the aspartokinase activity has been purified by chromatofocusing after limited proteolysis of aspartokinase I-homoserine dehydrogenase I from E.coli K12. The NH2-terminal sequence shows that it corresponds to the amino terminal peptide of the native enzyme. The results confirm a previous hypothesis about the organization of native aspartokinase I-homoserine dehydrogenase I. PMID- 2982667 TI - Comparative effects of cholera and Bordetella pertussis toxins on cyclic AMP and GTP levels and on lipolysis in rat adipocytes incubated in vitro. AB - The respective effects of cholera and Bordetella pertussis toxins were studied in time and concentration dependent experiments, following glycerol and fatty acid release, GTP and cAMP levels. Cholera toxin, after a lag time of 30 min, stimulated linearly GTP and cAMP accumulation and lipolysis (maximal effect: 2 fold increase at 5 micrograms/ml). Pertussis toxin presented a biphasic effect both in time and concentration dependent studies. Up to a maximum reached after 2 h with 1.4 units LPF/ml the stimulation affected GTP (3 fold) and cAMP (7 fold) levels, glycerol and fatty acid release (15 fold). Beyond this, an inhibition occurred, yielding a decrease towards basal values of GTP and cAMP content whereas the glycerol and fatty acid release was stopped. These results, which are the first reporting the fluctuation of the GTP content of intact cells challenged with bacterial toxins, show a close relationship between GTP and cyclic AMP levels and lipolytic activity. PMID- 2982666 TI - Molecular cloning and expression in Streptomyces lividans of a streptomycin 6 phosphotransferase gene from a streptomycin-producing microorganism. AB - The gene encoding streptomycin 6-kinase involved in the self-resistance of the streptomycin-producing Streptomyces griseus HUT 6037 was cloned in the plasmid vector pIJ703. The resulting plasmid, pSP6, contained 2.5 kb inserts of S. griseus DNA. When streptomycin-susceptible S. lividans 1326 was retransformed with pSP6, all transformants produced streptomycin 6-kinase. Addition of streptomycin to the culture medium of S. lividans carrying pSP6 plasmid brought about a remarkable increase in streptomycin 6-kinase activity in the cell extracts. It is suggested from the results that the production of streptomycin 6 kinase in streptomycin producer was induced by streptomycin accumulated during cultivation. PMID- 2982668 TI - Pertussis toxin facilitates the progesterone-induced maturation of Xenopus oocyte. Possible role of protein phosphorylation. AB - Progesterone triggers the first meiotic cell division of Xenopus oocyte and inhibits cAMP synthesis. The effect of pertussis toxin purified from Bordetella pertussis was tested on the maturation of Xenopus oocyte. The toxin did not inhibit progesterone-induced resumption of meiosis or the hormone-induced drop in cAMP level. This indicates that progesterone action is not mediated by the Ni subunit of the oocyte adenylate cyclase. Furthermore, pertussis toxin caused a reduction in the time course of maturation correlated with the precocious appearance of an alkali stable 47 kDa phosphoprotein, a marker of the maturation promoting factor (MPF) activity. Pertussis toxin effects mimicked those of 2 glycerophosphate suggesting that both agents act on the steady-state level of phosphorylation implicated in MPF activity. PMID- 2982669 TI - Calcium-dependent polyphosphoinositide hydrolysis is associated with exocytosis in vitro. AB - Micromolar calcium ions stimulate both exocytosis and polyphosphoinositide hydrolysis in sea urchin egg plasma membrane in vitro. Strontium and barium ions also stimulate both processes equally. Magnesium ions reduce the calcium sensitivity of both. Neomycin, a drug which prevents phosphoinositide hydrolysis, inhibits exocytosis in vitro. We suggest that hydrolysis of plasma membrane phosphoinositides may be an essential step in the fusion of the secretory granule and plasma membranes. PMID- 2982670 TI - Photolabelling of D-beta-hydroxybutyrate apodehydrogenase with azidoaryl phospholipids. AB - Two synthetic photoactive azidoarylphosphatidylcholines were used to investigate the level of interaction between D-beta-hydroxybutyrate apodehydrogenase (apoBDH), an amphipathic membrane protein, with the hydrophobic domain of phospholipids. The two synthetic lecithins, PL I (1-myristoyl-2-12-N-(4-azido-2 nitrophenyl) aminododecanoyl-sn-glycero-3-phosphocholine) and PL II (1-myristoyl 2-(2-azido-4-nitrobenzoyl)-sn-glycero-3-phosphocholine), are able to reactivate the non-active purified apoBDH as well as the non-photoactive homologs, indicating that the photoreactive chemical groups are without effect on the cofactor properties of phosphatidylcholine. Photoirradiation of reconstituted complexes between phospholipid containing azidoaryllecithin and apoBDH leads to a covalent binding of some synthetic lecithin molecules on the protein. The labelling, about 3 times higher with PL II than with PL I, suggests that the area of interacting domain of BDH with the hydrophobic moiety of phospholipid is more important at or near the surface of the lipid bilayer than in the inner part. This approach is further demonstration that BDH is an integral protein. PMID- 2982671 TI - Crosslinking of actin filaments is caused by caldesmon aggregates, but not by its dimers. AB - A recent report by Bretscher [(1984) J. Biol. Chem. 259, 12873-12880] showed that caldesmon prepared by his method crosslinks actin filaments to form thick bundles. This is in contrast to the results of previous work that caldesmon binds to F-actin but does not cause any gelation [(1981) Proc. Natl. Acad. Sci. USA 78, 5652-5655]. The present work clearly showed that caldesmon purified according to Bretscher does not cause any gelation of F-actin. However, caldesmon aggregates formed by concentration or by freeze-thawing gelated F-actin to form bundles. PMID- 2982672 TI - [Presynaptic autoregulation of dopamine release]. AB - Experiments in vivo with local perfusion of rat brain neostriatum revealed existence of autoregulation of dopamine release mediated by presynaptic dopamine autoreceptors. An increase in dopamine concentration in the perfusion medium (addition of exogenous dopamine 10(-7) M, or cocaine, the inhibitor of uptake of biogenous amines, 10(-6) M) leads to a decrease in K+--induced release into perfusate of 3H--dopamine preliminarily injected into neostriatum. Haloperidol (10(-6) M) abolishes the inhibitory effect of exogenous dopamine. Perfusion with dopamine--containing medium (10(-4) M) leads to a decrease in the amplitude of EPs recorded in the neostriatum upon electrical stimulation of zona compacta of substantia nigra, indicating that dopamine release from dopaminergic terminals of nigro--striatal neurons decreases in response to AP. PMID- 2982673 TI - [Participation of peptide mechanisms in physiological reactions of the cardiovascular system to hyperoxia]. AB - Experimental study in 156 white rats with renovascular hypertension, posthemorrhagic hypotension and with normal blood pressure revealed that high- ressure oxygen (303.9 kPa--50 min) could activate angiotensin--converting enzyme, the latter taking part in inactivation of endogenous opioid peptides. Involvement of peptide--like links in hemodynamic responses to hyperoxia seems to be an important component of the mechanism of oxygen action upon the cardiovascular system. PMID- 2982674 TI - A study of the role of macrophages in the graft-versus-host reaction. AB - Resting PEC and fractions of spleen cells adherent to nylon wool (NASC) or polystyrene (PASC) from A/Ph mice evoke a weak regional GVHR or no systemic GVHR in (B10 X A/Ph)F1 hybrids. Blockage of macrophages in vivo by silica or silica gels or removal in vitro of cells phagocytosing iron particles did not change GVH reactivity of A/Ph spleen cells. Addition of PEC or NASC from A/Ph mice to the A/Ph spleen cell suspension somewhat suppressed regional GVHR but enhanced systemic GVHR. Addition of PASC from A/Ph mice, conversely, enhanced regional GVHR and suppressed systemic GVHR. Repeated administration of silica and silica gel to the recipients suppressed regional GVHR; a single dose of silica was most effective at suppression when given one day before the cells. Systemic GVHR was not influenced by this treatment of the recipients. The results indicate that macrophages do not directly induce a GVHR, but they serve as important targets in regional GVHR. In regional and systemic GVHR macrophages perform regulatory functions. Macrophages of donor origin may modify both types of GVHR significantly, depending on the source and type of macrophages and the treatment of donors. PMID- 2982675 TI - Hormonal control of the phosphorylation of histones, HMG proteins and other nuclear proteins. AB - Hormone-dependent phosphorylation and dephosphorylation of nuclear proteins may play an important part in regulating nuclear function and specific gene expression. Some progress has been made in identifying specific nuclear proteins whose phosphorylation is affected by specific hormones; however, relatively little is known about the regulatory mechanisms involved, or about the molecular consequences of increased or decreased phosphorylation. It is suspected--but not yet proved--that cAMP-dependent effects on transcription are mediated at least partly by increases in nuclear cAMP-dependent protein kinase (A-kinase) activity, and consequent increases in the phosphorylation of specific chromatin proteins. In several instances, increased phosphorylation has been found to precede or correlate with cAMP-mediated induction of specific gene products. Several chromatin proteins are susceptible to cAMP-dependent phosphorylation in vivo, including histones H1 and H3, the high mobility group protein HMG 14 (which is preferentially associated with actively transcribed chromatin), and at least three other basic nonhistone proteins. The A-kinase phosphorylation sites of the majority of H1, H3 and HMG 14 molecules in chromatin appear to be inaccessible to A-kinase in vivo; nothing is known about the factors determining their accessibility, which may be tightly regulated and may vary significantly from cell to cell and tissue to tissue. Many hormone-induced changes in nuclear protein phosphorylation may be cAMP-independent. cAMP-independent mechanisms could involve a variety of nuclear enzymes including, for example, cGMP dependent, Ca2+/calmodulin-dependent, Ca2+/phospholipid-dependent and polyamine dependent protein kinases. So far, however, there is little solid evidence in support of a role for any specific cAMP-independent protein kinase in mediating hormonally induced increases in the phosphorylation of specific, identified nuclear proteins. PMID- 2982676 TI - Induction of prolactin receptors by prolactin in the rat lung and liver: demonstration of separate receptor and antibody entities. AB - This study examined the role of prolactin (PRL) in inducing its own receptors in rat lung and liver beyond the parallel immunological response evoked. Ovine PRL (oPRL), mixed with polyvinylpyrrolidone (PVP) and injected s.c. daily to male rats for 7, 10 and 14 days, was shown to induce specific binding of [125I]iodo oPRL in the lung and liver crude membrane fractions. Doses as low as 12.5 ng/kg were effective in inducing PRL-binding sites, which, however, differed qualitatively from those found in livers of 17 beta-estrogen-treated male rats. The oPRL-induced sites were highly specific for oPRL and were relatively stable to heat, suggesting the possible participation of antihormone antibodies in the binding observed. Indeed, anti-oPRL antibodies found in sera of oPRL-treated rats increasingly bound oPRL as a function of the duration of treatment. Water-washing of the membrane fraction succeeded in gradually eliminating the loosely bound antibody and in partially restoring the displacing ability of excess (1 micrograms) rat PRL on oPRL-binding sites in the lung (32.3%) and liver membranes (29.3%). Also restored were the heat lability typical of the receptor site, as well as the inhibitory effect of anti-PRL-receptor antiserum (1:100) on PRL binding (50-65% inhibition of total binding). In keeping with these results, in vitro incubation of liver membranes with rat anti-oPRL antiserum greatly reduced the ability of rat PRL to compete for oPRL binding, supporting the findings after in vivo treatment with oPRL.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982677 TI - Divergent responses by human and mouse thyroids to human chorionic gonadotropin in vitro. AB - hCG is a known stimulator of mouse thyroid in vivo. Studies were therefore performed to ascertain whether the thyroid-stimulating activity of hCG in the mouse could also be demonstrated by the in vitro techniques that had failed to show any activity of hCG in the human thyroid. When labeled with 125I and incubated at 22 degrees C in 20 mM Tris-0.5% bovine serum albumin (Tris-BSA), pH 7.45, with increasing concentrations (70-300 micrograms protein/ml) of a mouse thyroid fraction, a purified hCG preparation [( 125I]hCG) showed 5-12% specific binding. In contrast, its binding to a human thyroid particulate fraction, over the same range of protein concentrations, did not exceed 1%. When similar studies were performed at 37 degrees C in 10 mM Tris-50 mM NaCl-0.5% BSA, pH 7.45, [125I]hCG showed no detectable binding either to the human or the mouse thyroid fractions. At concentrations ranging from 1 to 20 mIU/ml (0.9-18 X 10(-9) M), bTSH stimulated cAMP release from human thyroid slices into the medium in a dose dependent manner. In contrast, hCG concentrations from 10(3) to 10(4) IU/ml (2-20 X 10(-6) M) were without effect on cAMP release. bTSH, at concentrations of 4.5 and 9.0 mIU/ml (4 and 8 X 10(-9)M), stimulated cAMP release from the mouse thyroid, producing in the medium approximately 11- and 28-fold increases in cAMP concentration. hCG also stimulated cAMP release from the mouse thyroid, the increases being approximately 2.3- and 1.8-fold, in the presence of 2270 and 4540 IU/ml (4.5 and 9.0 X 10(-6) M), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982678 TI - Calcium-cyclic AMP interactions in prothoracicotropic hormone stimulation of ecdysone synthesis. AB - The calcium-dependence of ecdysone synthesis by the insect prothoracic glands was examined in vitro using glands from day 0 pupae of the tobacco hornworm, Manduca sexta. Stimulation of ecdysone synthesis by the cerebral neuropeptide, prothoracicotropic hormone (PTTH), requires extracellular calcium; peptide stimulated steroidogenesis is blocked by omission of calcium or by addition of the calcium antagonist lanthanum. By contrast, basal synthesis of ecdysone is not calcium-dependent. A stimulatory, as opposed to simply a permissive, role for calcium is indicated by the ability of the calcium ionophore A23187 to mimic the steroidogenic effects of PTTH. Agents that act by increasing the intracellular levels of cAMP (dibutyryl cAMP, 1-methyl-3-isobutylxanthine, forskolin) enhance ecdysone synthesis equally well in the presence or absence of extracellular calcium, indicating that the site of action of the cation in the steroidogenic pathway occurs prior to that of cAMP. Both PTTH and A23187 enhance the formation of cAMP, as measured by the conversion of [3H]ATP to [3H]cAMP, in a manner absolutely dependent upon the presence of extracellular calcium. The results suggest sequential roles for calcium and cAMP in PTTH-stimulated steroidogenesis by the insect prothoracic glands. A model is presented in which the peptide stimulates cAMP formation in a calcium-dependent manner, with the cyclic nucleotide in turn enhancing ecdysone synthesis. PMID- 2982679 TI - Cyclic AMP levels during the maturation of Xenopus oocytes. AB - While several studies have suggested that the induction of oocyte maturation results from a transient decrease in cAMP levels, attempts to demonstrate such a change have led to inconsistent results with respect to whether or not a decrease occurs as well as timing of the decrease. In this report the results of experiments designed to demonstrate small changes in cAMP content in Xenopus laevis oocytes are presented and a statistically significant 20% decrease in cAMP content from between 2 and 50 min postprogesterone addition is found. The cAMP content subsequently rises to 12% higher than control levels and then becomes indistinguishable from control values for the remainder of the maturation period. PMID- 2982680 TI - Inhibition by dibutyryl cyclic AMP of the transition to metaphase of mouse oocyte nuclei and its reversal by cell fusion to metaphase oocytes. AB - Mouse oocytes at metaphase I of meiotic maturation were treated with puromycin, which caused the condensed chromosomes to become decondensed to form an interphase nucleus. The chromosomes returned to a metaphase state 6.3 hr after the oocytes were transferred to puromycin-free medium [H. J. Clarke and Y. Masui (1983) Dev. Biol. 97, 291-301]. In contrast, the chromosomes of the puromycin treated oocytes remained decondensed within the nucleus if dibutyryl cyclic AMP (dbcAMP) was included in the puromycin-free medium. This implies that dbcAMP inhibited the development of conditions in the oocytes that were required for the transition to metaphase. The chromosomes of puromycin-treated oocytes that were incubated for 7.5 hr in dbcAMP-containing medium returned to metaphase just 1.9 hr after transfer to dbcAMP-free medium. Therefore, the protein synthesis dependent process that is required for the transition to metaphase could occur in the presence of dbcAMP. Fusion to metaphase II oocytes, or to puromycin-treated oocytes that had returned to metaphase, rapidly induced transition of the nuclei of dbcAMP-inhibited oocytes to metaphase, despite the presence of the inhibitor. These results suggest that the transition of nuclei to metaphase can be induced by a cytoplasmic factor that is present in metaphase oocytes, and that dbcAMP inhibits the development of this factor. PMID- 2982681 TI - Nonenzymatic glycosylation of erythrocytic proteins in normal and diabetic subjects. Enzymes of nucleoside and nucleotide metabolism. AB - A modification of the technique of Glyco-Gel affinity column chromatography has been employed to separate glycosylated proteins from nonglycosylated proteins of hemolysates. When glycosylation in hemolysates of 11 type I diabetic subjects was compared with that from 7 normal subjects, significant increases were found in glycosylation of hemoglobin (Hb) (12.1 +/- 6.0% versus 4.7 +/- 0.5%) and purine nucleoside phosphorylase (PNP) (5.3 +/- 3.0% versus 2.1 +/- 0.5%). However, no differences were found for nucleoside diphosphokinase (NDPK) (1.5 +/- 1.1% versus 1.0 +/- 0.4%) and adenylate kinase (AMPK) (0.5 +/- 0.4% versus 0.7 +/- 0.2%). Linear relationships were seen between glycosylated Hb and glycosylated PNP (r = 0.97) or glycosylated NDPK (r = 0.81). On incubation of hemolysates from normal individuals with high glucose (1500 mg/dl or 83 mM) and NaCNBH3 (20 mM), linear increases in the degrees of glycosylation were seen with time. After 18 h, the percentages of glycosylation of Hb, PNP, NDPK, and AMPK were increased from normal values to 31, 24, 11, and 3, respectively. When partially purified human erythrocytic PNP was incubated with various monosaccharides (20 mM) in the presence of NaCNBH3 for 6 h, glycosylation increases of 2-5-fold were seen in the order ribose greater than mannose greater than galactose greater than glucose. PMID- 2982682 TI - Effect of E-series prostaglandins on cyclic AMP-dependent and -independent hormone-stimulated glycogenolysis in hepatocytes. AB - The effect of E-series prostaglandins (PGE) on hormone-stimulated glycogenolysis was studied in isolated rat hepatocytes. As previously reported, the physiologically active analogue 16,16-dimethyl-PGE2 inhibited glucagon-stimulated glycogenolysis. This effect could be reproduced by repetitive addition of PGE2 to compensate for PGE2 catabolism. In contrast, glycogenolysis stimulated by N6,O2' dibutyryladenosine-3',5'-cyclic monophosphate (dibutyryl-cAMP) was unaffected by either PGE2 or 16,16-dimethyl-PGE2 (rate of glycogenolysis with 0.34 microM dibutyryl-cAMP plus 1.7 microM 16,16-dimethyl-PGE2 = 99 +/- 6% of rate with 0.34 microM dibutyryl-cAMP alone; mean +/- SEM, N = 5). Similarly, glycogenolysis stimulated by 8-bromoadenosine-3',5'-cyclic monophosphate was not inhibited by PGE2 or 16,16-dimethyl-PGE2. Epinephrine-stimulated glycogenolysis was inhibited by 16,16-dimethyl-PGE2 in a dose-dependent manner. PGE inhibited the cAMP independent stimulation of glycogenolysis resulting from phenylephrine or angiotensin II exposure (rate of glycogenolysis with 8 microM phenylephrine + 1.7 microM 16,16-dimethyl-PGE2 = 65 +/- 10% of rate with 8 microM phenylephrine alone, N = 4, P less than 0.05; 4.9 microM angiotensin II + 1.7 microM 16,16 dimethyl-PGE2 = 75 +/- 7% of rate with 4.9 microM angiotensin II alone, N = 4, P less than 0.05). Glycogenolysis stimulated by the calcium ionophore A23187 was also inhibited by PGE (rate of glycogenolysis with 0.55 micrograms/ml A23187 + 1.7 microM 16,16-dimethyl-PGE2 = 83 +/- 5% of rate with 0.55 micrograms/ml A23187 alone, N = 7, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982683 TI - Islet beta-cell function and polymorphism in the 5'-flanking region of the human insulin gene. AB - The present study investigates the possible relationship between human beta-cell secretory capacity and polymorphism in the 5'-flanking region of the human insulin gene. The glucose potentiation slope was measured in normal and non insulin-dependent diabetic subjects (NIDDM). This slope, as reported previously (Ward, W. K., et al., Am. J. Physiol. 1984; 246:E405-11), is an index of the ability of hyperglycemia to potentiate the insulin response to arginine and as such is a measure of beta-cell responsiveness to glucose. Restriction enzyme analysis using a human insulin gene probe was performed on leukocyte DNA isolated from the same individuals. We conclude that a 1.6 kb polymorphism in the 5' flanking region of the human insulin gene in both normal and NIDDM subjects has no association with insulin secretory responses as defined here by the glucose potentiation slope. PMID- 2982684 TI - Demonstration that the vitamin D metabolite 1,25(OH)2-vitamin D3 and not 24R,25(OH)2-vitamin D3 is essential for normal insulin secretion in the perfused rat pancreas. AB - It has previously been shown that vitamin D deficiency impairs arginine-induced insulin secretion from the isolated, perfused rat pancreas (Science 1980; 209:823 25). Since vitamin D is known to be metabolized to 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) and 24R,25-dihydroxyvitamin D3 (24,25[OH]2D3), it is essential to clarify which vitamin D metabolite has the important role of enhancing insulin secretion. In this report, a comparison is made of the relative efficacy of 3-wk repletion with vitamin D3 (980 pmol/day), 1,25(OH)2D3 (39 pmol/day or 195 pmol/day), and 24,25(OH)2D3 (650 pmol/day) on arginine-induced insulin secretion from the isolated, perfused rat pancreas; in this experiment, the daily caloric intake of the animals receiving vitamin D or its metabolites was controlled by pair feeding to the caloric intake of the vitamin D-deficient rats. 1,25(OH)2D3 repletion was found to completely restore insulin secretion to the levels seen in vitamin D3-replete, pair-fed controls in both the first and second phases, while 24R,25(OH)2D3 only partially improved insulin secretion, and then only in the first phase. Changes of both serum calcium levels and dietary caloric intake after vitamin D metabolite administration are concluded to play a lesser role on the enhancement of insulin secretion, since, in a separate experiment, vitamin D deficient rats with normal serum calcium levels did not show recovery of insulin secretion equivalent to the vitamin D-replete animals under conditions of dietary pair feeding. These results suggest that 1,25(OH)2D3 but not 24,25(OH)2D3 plays an essential role in the normal insulin secretion irrespective of the dietary caloric intake and prevailing serum calcium levels. PMID- 2982685 TI - Localization of saturable CCK binding sites in rat pancreatic islets by light and electron microscope autoradiography. AB - Cholecystokinin (CCK) is a known stimulus for the release of insulin and other islet hormones. To localize islet cell CCK binding sites, we measured the uptake of 125I-CCK by the isolated, perfused rat pancreas. Light microscope autoradiographs revealed uptake of label over both the endocrine islets of Langerhans and the exocrine acini. This uptake of 125I-CCK was saturable, as it decreased markedly when a large excess of unlabeled CCK8 was included in the perfusion solution. To define which cells in the islets bound CCK, electron microscope autoradiographs were prepared. The majority of silver grains in islets were localized over beta cells (69%), although saturable uptake was also observed over alpha (12%) and other islet cells. When grain densities were analyzed (grains/micron 2), the highest density was observed over islet blood vessel cells. In contrast to islet blood vessels, there was no localization of 125I-CCK over acinar blood vessels. This study supports the concept, therefore, that there is a direct regulation of islet endocrine cells by CCK, and also raises the possibility that CCK influences islet hormone release via an indirect effect on the islet vascular endothelium. PMID- 2982686 TI - Age-related changes in hepatic glycogen metabolism in the genetically diabetic (db/db) mouse. AB - Hepatic glycogen metabolism was investigated in genetically diabetic C57BL/KsJ db/db mice during their development. Initially, the development of obesity, hyperglycemia, hyperinsulinemia, and hyperglucagonemia in these mice was examined, which illustrated that the diabetes progressed normally. Little difference in hepatic glycogen concentrations was observed, averaging approximately 50 and 60 mg/g liver in diabetic (db/db) and control heterozygote (db/+) mice, respectively. Glycogen synthase activity (total and a-form) was significantly elevated by 5 wk in the diabetic mice relative to controls and reached maximum levels (two-fold higher than controls) around 8-9 wk. This activity then slowly declined during the rest of the 15-wk period examined. Both phosphorylase a and total phosphorylase activities were also elevated by 5 wk, reaching levels twofold higher than controls. These activities did not decline at the end of this 15-wk period, but instead continued to slowly increase. Glycogen synthase a activity showed a positive correlation (r = 0.54, N = 144) with circulating levels of insulin, and a similar correlation was seen for phosphorylase a activity and plasma glucagon levels (r = 0.64, N = 72). Protein kinase and phosphoprotein phosphatase activities were also measured, but no differences were detected between diabetic and control mice. This longitudinal study clarifies some of the changes in hepatic glycogen metabolism that occur during the progression of diabetes in the db/db mouse and indicates a role for circulating insulin and glucagon concentrations on the steady-state activities of glycogen synthase and phosphorylase, respectively. PMID- 2982687 TI - [Acquired immunodeficiency]. PMID- 2982688 TI - [Current status and predictions in gynecology and obstetrics]. PMID- 2982689 TI - Methylation pattern of the bovine leukemia provirus genome in bovine leukemic cells. AB - All the known genes of the bovine leukemia proviral genome were found to be heavily methylated in both fresh and short-term-cultured leukemic cells, the latter of which expressed viral antigens. However, the pXBL region appeared to be mostly demethylated or at least hypomethylated in both cells, indicating that there is a non-uniform methylation pattern of the proviral genome. PMID- 2982690 TI - Seroepidemiology of adult T-cell leukemia virus in Taiwan. AB - A survey of carriers of adult T-cell leukemia virus (ATLV) detected as anti-ATLV associated antigens was made in Taiwan. Among 2545 adults aged 30 years or more examined, seropositive donors amounted to 0.9% in the Han Chinese but none in the aborigines. PMID- 2982691 TI - Granular cell tumors of the esophagus. AB - An unusual benign multicentric esophageal granular cell tumor (granular cell myoblastoma) associated with 16 other similar tumors in the skin, vulva, breast, and tongue of 1 patient is described. There was a family history of granular cell tumors in the patient's mother; this has not been previously described. The pathologic findings and controversial histogenesis of granular cell tumors are discussed in an effort to delete the erroneous term "myoblastoma" from the radiologist's vocabulary. Granular cell tumors of the esophagus are also specifically reviewed. PMID- 2982692 TI - Pseudotumorous pancreatitis. AB - Chronic pancreatitis can occasionally present as a focal, noncalcified mass, indistinguishable from carcinoma. Radiologic studies in 21 such patients were considered to show carcinoma in 16 and an islet cell tumor in 5 patients. Seventeen of the patients came to laparotomy when a palpable mass was found in each; 9 of the patients had a partial pancreatectomy, and multiple biopsy specimens were taken in 8 patients. Four patients did not come to surgery; in them the diagnosis was based on examination of percutaneous biopsy specimens and follow-up. In all patients the histologic diagnosis was chronic relapsing pancreatitis. PMID- 2982693 TI - How does cobalamin (vitamin B12) enter and traverse the ileal cell? PMID- 2982694 TI - Beneficial effect of vegetable protein diet supplemented with psyllium plantago in patients with hepatic encephalopathy and diabetes mellitus. AB - A controlled crossover study was performed in 8 diabetic patients with chronic portal-systemic encephalopathy. After a basal period the patients were treated during periods A and B. During period A, a meat protein diet (0.8 g/kg body wt, approximately 1800 kcal/day) was consumed and neomycin plus laxatives were given. During period B patients received vegetable protein (0.8 g/kg body wt, 1800 kcal/day). This diet was supplemented with psyllium fiber to reach 35 g of fiber per day. Four patients were randomly assigned to receive the treatments in the order A-B and the other 4 patients in the order B-A. At the end of the first experimental period, fasting glucose levels were 204 +/- 86 mg% in the meat protein diet group and 127 +/- 8 mg% in the vegetable protein diet group (p less than 0.014). The patients were receiving 2.5 +/- 0.2 g/day and 2.1 +/- 0.5 g/day of tolbutamide at the end of the meat protein diet and vegetable protein diet, respectively. In all cases, fasting glucose levels decreased at the end of the vegetable diet period regardless of the previous treatment. An improvement of greater than or equal to 25 mg% of fasting glucose levels was observed in 7 of the 8 patients after the vegetable protein diet and in no case after the meat protein diet (p less than 0.0078). The parameters of encephalopathy were comparable at the end of both the meat protein diet and the vegetable protein diet. A significant increase in the number of bowel movements was noticed after the vegetable diet plus fiber (p less than 0.01). We propose the use of vegetable diet plus fiber to facilitate the treatment of patients with both diabetes and hepatic encephalopathy. PMID- 2982695 TI - In vivo evidence that cobalamin is absorbed by receptor-mediated endocytosis in the mouse. AB - This study examines the mechanism of cobalamin absorption in the context of receptor-mediated endocytosis. Uptake, the amount of cobalamin that left the intestinal lumen, and transport, the component that was located in organs beyond the intestine, were measured after feeding saturating doses of [57Co]cobalamin to mice. Uptake from a 40-ng dose of [57Co]cobalamin at 1 h was 18.6 +/- 6.3 ng (mean +/- SD; n = 6). When the dose was given 1 h after 40 ng of unlabeled cobalamin, uptake was 18.5 +/- 1.5 ng, n = 6, indicating rapid clearance of the surface receptors. Transport of the initial and of a subsequent dose of cobalamin could not be detected for 2 h but the amounts and rates of transport were similar. Oral chloroquine and intraperitoneal cycloheximide reduced transport at 4 h to 4.3 +/- 1.6 ng, n = 8 and 5.6 +/- 2.4 ng, n = 7, respectively. Control values were 14.3 +/- 1.3 ng, n = 8. These results indicate that the transport of cobalamin involves a series of compartments, one of which may be lysosomal, and that there is a requirement for new protein synthesis. PMID- 2982696 TI - Antibiotic depression of evoked and spontaneous responses of opossum distal colonic muscularis mucosae in vitro: a factor in antibiotic-associated colitis? AB - Certain antibiotics depress both skeletal neuromuscular transmission and intestinal neuroeffector transmission. Impaired intestinal motility may facilitate the proliferation of the bacterium Clostridium difficle and thus lead to the development of antibiotic-associated pseudomembranous colitis. Many antibiotics accumulate in the colonic lumen at concentrations several times their associated blood levels. This study examined whether certain of these could interfere with colonic muscularis mucosal movement in vitro, using tissue from opossum distal colon as a model. At concentrations approximating those in the colonic lumen, ampicillin, clindamycin, erythromycin, and lincomycin depressed tone and spontaneous contractions of the muscularis mucosae. Clindamycin, gentamicin, kanamycin, and lincomycin abolished electrically evoked contractions but only gentamicin and kanamycin could abolish the ensuing relaxation. Vancomycin potentiated the response of the muscularis mucosae to acetylcholine; erythromycin and clindamycin depressed it. Antibiotic-induced depression of colonic muscularis mucosal movement may contribute to the development of antibiotic-associated pseudomembranous colitis. PMID- 2982698 TI - The development of ACTH-like substance during tadpole metamorphosis. AB - The interrenals of tadpoles of Rana catesbeiana at various developmental TK stages were frozen, sectioned, and prepared histochemically for assessment of delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD) activity. The interrenals of hypophysectomized (HX) tadpoles from the same batch of eggs as control tadpoles were prepared similarly for delta 5-3 beta-HSD activity following a series of intraperitoneal injections of different concentrations of mammalian ACTH. The histochemically stained sections of control and HX tadpoles were scanned with a computerized microscope spectrophotometer for diformazan absorbance. Two regression equations were obtained: Y = 0.1460 + 0.0015X and Y' = 0.1494 + 0.0893X', where Y or Y' = delta 5-3 beta-HSD activity absorbance (for the control and HX tadpoles, respectively), X = developmental stages of control, and X' = exogenous ACTH dosage administered to the HX tadpoles. Taking absorbance of delta 5-3 beta-HSD activity as the common factor, a curve of concentrations of ACTH-like substance against TK stages was made. The developing concentration of ACTH-like substance was very low at first and then rose during metamorphic climax. The trend of the development of ACTH-like substance was discussed from the view point of thyroid and delta 5-3 beta-HSD activities. PMID- 2982697 TI - Gonadotrophin-induced oocyte maturation in the catfish, Heteropneustes fossilis (Bloch), requires steroidogenesis in both interrenal and ovary. AB - Intact or hypophysectomized catfish, Heteropneustes fossilis, were administered a single injection of ovulating doses of ovine luteinizing hormone (LH: 200 micrograms/100 g body wt) or partially-purified salmon gonadotrophin (SG-G100: 100 micrograms/100 g body wt). Identical groups of catfish were injected with a suboptimal dose of LH (20 micrograms/100 g body wt) or with porcine adrenocorticotrophin (ACTH: 0.25 IU/100 g body wt). At short intervals after hormone administration, plasma and/or ovarian tissue were analyzed for cortisol (F), testosterone (T), and estradiol-17 beta (E2) by radioimmunoassay. Following administration of ovulatory doses of gonadotrophins, plasma levels of the three steroids increased in a sequential manner; high levels were recorded between 15 and 45 min for F and between 45 and 90 min for T and E2. In gonadotrophin injected catfish, the ovarian content of T and E2 increased during the first 45 min and then declined up to 90 min even as their titers in the plasma were still increasing. When ovarian pieces containing yolky oocytes were incubated in vitro with LH (50 micrograms/ml), levels of T and E2 in the culture medium increased in a sequential manner similar to that observed following in vivo administration of gonadotrophin. No significant change was observed in the levels of any of the three steroids in catfish injected with a suboptimal dose of LH. In catfish treated with ACTH, plasma F levels increased 40-fold, whereas T and E2 levels did not change; ACTH administration had no effect on oocyte maturation. These results suggest that gonadotrophin, at doses sufficient to evoke oocyte maturation, acts at two loci, the interrenal and the ovary. The results also suggest that the failure of ACTH to induce oocyte maturation is due to its inability to act on the ovary. PMID- 2982700 TI - Evolution and extinction of transposable elements in Mendelian populations. AB - A model of the evolution of a transposable element family in a Mendelian host population is proposed that incorporates heritable phenotypic mutations in the elements. The temporal behavior of the numbers of mutant and wild-type elements is studied, and the expected extinction time of the transposable element family is examined. Our results indicate that, if the mutant can be transposed equally well in the presence of the wild type, then it can be expected to be found in preponderance, whereas elements, such as retroviruses, where the transposing genome and its phenotypic expression are coupled, may be characterized by a low mutant frequency. PMID- 2982699 TI - Molecular analysis of X-ray-induced alcohol dehydrogenase (ADH) null mutations in Drosophila melanogaster. AB - We have attempted to analyze at the molecular level mutants previously determined as having intragenic lesions caused by X-ray mutagenesis. C.S. Aaron isolated 33 null mutations at the Adh locus and in collaboration with other investigators classified 23 as deletions. Of the eight mutants analyzed here, only two produced a detectable ADH protein using the two-dimensional electrophoresis technique. Restriction endonuclease and Southern blot analysis showed that three of the mutants were normal compared to the wild-type restriction pattern, with one of the three producing a mutant ADH protein. Among the five mutants that had altered restriction patterns, only one mutant produced a detectable mutant ADH protein. All the mutants produced a hybridizable mRNA when probed with the genomic clones, suggesting that the mutant phenotype was not due to transcriptional inhibition. Two probable explanations proposed for these observations are (1) mutations may be due to deletions of one or a few bases resulting in frameshifts to nonsense codons and premature termination of ADH peptide synthesis or (2) mutations may be a result of transitions to nonsense codons, again producing shortened ADH proteins. Those mutants producing a mutant polypeptide may have resulted from mutations to missense rather than nonsense codons. The five mutants showing an abnormal endonuclease Southern blot along with the 23 mutants previously shown to be deletions (28/33 or 85%) are associated with multiple DNA chain breaks. Although all of the DNA chain breaks are not necessarily associated with the mutant phenotype of the Adh locus, multiple DNA chain breaks are the most consistent characteristic of ionizing radiation damage to DNA. PMID- 2982701 TI - [Characteristics of the clinical picture and course of neurotic disorders in people engaged in diamond cutting]. PMID- 2982702 TI - Squamous cell carcinoma of the cervix stage IB: numbers and reactivities of pelvic lymph node T cells. AB - Ten patients with squamous cell carcinoma of the cervix stage IB (FIGO) treated by radical hysterectomy and pelvic lymph node dissection had node biopsies taken for immunological studies. There was no evidence of metastases. Node biopsies from near the cervix (the obturator region) contained significantly more lymphoid cells per gram of tissue than biopsies from more distant (common iliac) nodes. The percentage of T cells was similar in the two nodes, but significantly lower than in mononuclear cell suspensions from peripheral blood. All patients responded to the mitogen phytohemagglutinin. Positive T-cell responses to herpes simplex virus antigen were found in seven patients. Six patients responded to stimulation by the chlamydial antigen LGV-2. Lymph node T cells gave responses higher than those from peripheral blood T cells. Obturator node T cells from patients pretreated with intracavitary radium had antigen-specific responses lower than those of the patients operated without prior irradiation. The results indicate that the pelvic lymph nodes are important reservoirs for sensitized T cells. PMID- 2982703 TI - Cell-mediated and humoral immune responses to chlamydial and herpesvirus antigens in patients with cervical carcinoma. AB - Herpes simplex virus (HSV) and Chlamydia trachomatis are both discussed in the etiology of cervical carcinoma. In this study the antibody titers and the T-cell proliferative responses to chlamydial and HSV antigens in patients with cervical intraepithelial neoplasia (CIN) and invasive cervical cancer, have been investigated and compared. The patients with CIN and invasive cancer showed approximately the same degree of immune responses to chlamydial and HSV antigens. Of the patients, 58% showed proliferative T-cell responses to C. trachomatis antigen, 87% to HSV antigens. Chlamydial antibodies were detected in 65% of the patients, while 81% had a positive HSV serology. Our results show a lack of correlation between levels of antibody titer and T-cell responses. It is concluded that patients with CIN and invasive cervical cancer have intact cellular immune responses to both chlamydial and HSV antigens. The eventual role of these infections in the etiology remains unclear. PMID- 2982704 TI - Sarcoma-like mural nodules combined with a microfocus of anaplastic carcinoma in mucinous ovarian tumor. AB - A case of mucinous ovarian tumor with sarcoma-like mural nodules, one of which coexisted with a microfocus of anaplastic carcinoma, was studied by light, electron microscopic, and histochemical examinations. The case suggested that the formation of sarcoma-like mural nodules may be the result of the proliferation of undifferentiated mesenchymal cells which exist beneath the mucinous epithelium by some stimulation like hemorrhage in the cyst wall. Histologically, the mononucleated cells composing the sarcoma-like mural nodules had epithelioid characteristics, particularly at the areas where the covering mucinous epithelium had already detached. The case also suggested that the findings of epithelioid characteristics in the sarcoma-like mural nodules themselves do not imply these nodules to be anaplastic carcinoma, but merely an epithelioid differentiation from undifferentiated mesenchymal cells, namely one of reactive products. However, the existence of a microfocus of anaplastic carcinoma combined with the sarcoma-like mural nodule necessitates a careful histologic analysis of mural nodules for the treatment of patients and the determination of the prognosis. PMID- 2982705 TI - Cisplatin, methotrexate, and 5-fluorouracil combination chemotherapy for advanced ovarian cancer. AB - A combination chemotherapy including cisplatin, 25 mg/m2 on Days 1,8; methotrexate, 30 mg/m2 on Day 1; and 5-fluorouracil, 600 mg/m2 on Day 1 has been evaluated in 28 previously untreated and 10 pretreated patients with advanced ovarian cancer after debulking surgery when feasible. The pathological response rates (complete + partial responses) were 69.2 and 50% in untreated and pretreated patients, respectively. Overall 24-month survival and progression-free survival (PFS) are 19.2 and 10.9%, respectively. A significant difference in survival and PFS is evident between patients with less and more than 2 cm residual disease and between responders (CR + PR) versus nonresponders. No renal toxicity was induced and no cycles had to be delayed because of hemathologic toxicity. PMID- 2982706 TI - Renal metastases of malignant gestational trophoblastic disease: the use of intravenous urography in staging. AB - Intravenous urography (IVU) was performed in 289 patients at the time of diagnosis of malignant gestational trophoblastic disease (GTD). Ninety-five percent of these studies were normal, and none of the urologic abnormalities detected provoked alteration of the patients' tumor-related therapy. In the detection of renal metastases, false-positive and false-negative intravenous urograms were encountered. It is concluded that IVU lacks sufficient sensitivity and specificity in the detection of renal metastatic GTD; alternate screening methods are recommended. PMID- 2982707 TI - Uterine sarcoma in Israel: a study of 104 cases. AB - Data gathered during the first epidemiologic study of sarcoma of the uterus in Israel are presented. In the 7-year period of this survey, 104 new cases of sarcoma of the uterus were diagnosed, representing an incidence of 1.55/100,000 females over the age of 20. Half of the patients were in their fifth and sixth decades of life at the diagnosis of the tumor. Uterine sarcomas were more prevalent in women of European-American origin than in those of Asian-African descent. In 54.9% of the patients the diagnosis was made while the disease was in stage I. A definite correlation between sarcoma of the uterus, diabetes, hypertension, and obesity was not found. The 5-year survival rate in the present survey was 38%. Factors affecting prognosis were the clinical stage of the disease at diagnosis, histologic type, and the method of treatment. PMID- 2982708 TI - [New knowledge of the therapy and prevention of liver diseases. Key points: hepatitis vaccinations--differential diagnosis of hepatitis]. PMID- 2982709 TI - [An encouraging balance. Technology of obstetrics--embryo transfer--difficulty of tumor therapy--diagnosis, after care and education]. PMID- 2982711 TI - [Glomus tumors of the skin. Histologic subtypes, localization and clinical aspects]. PMID- 2982710 TI - [Progress in cardiologic therapy. Thrombolysis expanded--prevention possibilities employed early--ACE inhibitors as therapeutic hit]. PMID- 2982712 TI - Nuclear thyroxine receptors and cellular metabolism of thyroxine in obese subjects before and after fasting. AB - Nuclear binding of thyroxine (T4), cellular deiodination of T4 and nuclear accumulation of endogenous triiodothyronine (T3) were investigated in mononuclear blood cells in 5 obese persons before and after 7 days of total fast. Serum-free T4, serum-free T3 and serum thyrotropin (TSH) did not differ between lean persons and obese fed subjects, but fasting induced a significant decrease of serum-free T4 and serum-free T3 (T4 from 16.6 to 14.4 pmol/l, T3 from 5.7 to 3.2 pmol/l). The maximal specific binding capacity (MBC) for T4 was decreased in fed obese subjects (MBC = 0.6 fmol T4/100 micrograms DNA, p less than 0.05) compared to normal weight persons (MBC = 1.3 fmol T4/100 micrograms DNA), whereas the equilibrium association constant (ka) did not differ. During complete fast, the MBC increased (MBC = 1.5 fmol T4/100 micrograms DNA, p less than 0.05). Neither total deiodination of T4 nor endogenous nuclear T3 accumulation differed between the groups suggesting that cellular T4 to T3 conversion is independent of the nutritional state. We suggest that the observed increase in nuclear T4 receptor capacity may compensate the decrease in serum thyroid hormone levels. PMID- 2982713 TI - Metastatic and independent cancers of the endometrium and ovary: a clinicopathologic study of 34 cases. AB - Twenty-one of 34 simultaneous cancers involving the endometrium and ovary were classified as endometrial primary tumors with ovarian metastases. The criteria for this classification were either a multinodular ovarian pattern (major criterion) or two or more of the following minor criteria: small (less than 5 cm) ovary(ies), bilateral ovarian involvement, deep myometrial invasion, vascular invasion, and tubal lumen involvement. Twelve cancers were classified as independent neoplasms, primarily by the absence of the above criteria. Although they were classified as independent, the histologic features of the endometrial and ovarian tumors were the same in 11 of these 12 cases. Only one case represented an ovarian primary tumor with an endometrial metastasis. Both the group believed to have endometrial primaries with ovarian metastases and that with independent primaries showed high incidences of associated endometrial hyperplasia, supporting the belief that the endometrium is a primary site in both groups. The cancers classified as metastatic, with no known spread outside the endometrium-myometrium and ovary, were found to involve other sites significantly (P less than 0.01) more frequently than those classified as independent. Grade 3 endometrioid carcinoma, adenosquamous carcinoma, and malignant mixed mullerian tumors occurred only in the metastatic group, whereas the independent group had a variety of endometrioid and nonendometrioid tumors. PMID- 2982714 TI - A distinctive glomerular lesion complicating placental site trophoblastic tumor: report of two cases. AB - Renal disease with distinctive pathologic features developed in two young women who had placental site trophoblastic tumors. The renal abnormalities were manifested by proteinuria in both cases and by hematuria in one case; blood pressure was elevated in one of the patients. Pathologic examination of the kidneys showed distinctive glomerular abnormalities, characterized mainly by the presence of occlusive eosinophilic deposits in many of the glomerular capillary lumina, most of which stained for fibrinogen-related antigens and IgM by immunohistochemical techniques. Ultrastructural examination showed the deposits to consist mainly of granular material that contained packets of fibrillar material with the appearance of fibrin. The uterine tumors were composed of mononucleated and multinucleated cells with abundant cytoplasm that infiltrated between the muscle bundles of the myometrium; in both tumors there was prominent deposition of eosinophilic material that had the tinctorial properties of fibrin and that stained for fibrinogen and IgM in immunoperoxidase studies. The renal abnormalities disappeared after hysterectomy in one case; the other patient, who was receiving chemotherapy and had disseminated intravascular coagulation, died with leukopenia and sepsis. The clinical and pathologic features in these cases suggest that the renal abnormalities were related to the uterine tumors and that the production of immune complexes and/or the activation of intravascular coagulation by the tumors were pathogenetic mechanisms. PMID- 2982715 TI - An ultrastructural comparison of mesotheliomas with adenocarcinomas of the lung and breast. AB - The ultrastructural features of 15 mesotheliomas were compared with those of equal numbers of adenocarcinomas of the lung and of the breast in a double-blind study. Combined quantitative and qualitative features were evaluated to provide criteria for distinguishing among these three tumors, which may present as either primary or metastatic pleural tumors. mesotheliomas could be distinguished from adenocarcinomas of the lung by length of microvilli (mean ratios of length to diameter [LDR], 15.7 and 8.7, respectively; P less than 0.01) and content of tonofilaments. Length of microvilli was also useful in distinguishing mesotheliomas from breast adenocarcinomas (mean LDR, 15.7 and 6.9, respectively; P less than 0.001). Adenocarcinomas of the lung could be distinguished from adenocarcinomas of the breast by tonofilament content and the presence of intracytoplasmic lumina. Combined quantitative and qualitative criteria are essential for maximal ultrastructural discrimination among these tumors. PMID- 2982716 TI - Extragonadal germ cell tumors of the head and neck region: review of 16 cases. AB - During the period from 1928 to 1982, 16 children were treated or seen in consultation at the Children's Hospital Medical Center for extragonadal germ cell tumors of the head and neck region. Collectively, these tumors accounted for only 5 per cent of all benign and malignant germ cell tumors. Fourteen tumors were diagnosed in newborns and were classified as pure teratomas, either mature or immature. Six of these tumors were located in the cervical region, three presented within superficial facial structures, two were retro-orbital, and three arose within oropharyngeal or nasopharyngeal tissues. There were three endodermal sinus tumors, one of which appeared in an 11-month-old child who had undergone incomplete resection of an oropharyngeal teratoma as a newborn. Endodermal sinus tumors presented within the oropharynx, nasopharynx, and floor of the mouth and affected children at older ages (6 to 11 months). The overall prognosis for infantile teratomas of the head and neck region (exclusive of brain and spinal cord) is excellent, despite the presence of immature elements; however, tumor related deaths can result from large unresectable primary tumors. The prognosis for children with endodermal sinus tumors remains guarded, with successful management depending on early diagnosis and aggressive adjuvant therapy. PMID- 2982717 TI - Clinical trials of interleukin-2 in the acquired immune deficiency syndrome: warning to autopsy prosectors and other health care professionals. PMID- 2982718 TI - Observer reproducibility during computer-assisted planimetric measurements of nuclear features. AB - To evaluate observer variance during microprocessor-assisted planimetry, nuclear features (area, perimeter, and form factor) were studied in a series of mammary ductal carcinomas. Fragments of formalin-fixed, paraffin-embedded tissue were re embedded in plastic, sectioned at 1 micron, stained with methylene blue, and studied with a Zeiss-Kontron MOP-3 microprocessor-assisted planimeter. Both interobserver and intraobserver reproducibility were evaluated, the former among three different observers and the latter by two observers repeating their own measurements. Reproducibility was good for measurement of area, but deteriorated progressively for measurement of perimeter and form factor. Not only was observer correlation poor (identified via linear regression), but paired t-tests also showed consistent variation among observers. The major difficulty was in following the irregular nuclear contours that are characteristic of cancer cells. It is concluded that adequate demonstration of observer reproducibility remains an essential part of tissue investigation, even when the objective methods typical of morphometry are used. PMID- 2982719 TI - Subclassification of small cell cancer of the lung: the Southeastern Cancer Study Group experience. AB - Cancers in more than 300 patients being treated by Southeastern Cancer Study Group protocols for small cell carcinoma of the lung (SCCL) were subclassified as oat cell or intermediate types by criteria developed by two groups of investigators. With Cox survival and logistic regression models, no differences in survival or responsiveness to therapy were found between these types of tumors. These and previously published results suggest that the only intermediate tumors worth distinguishing are the "22/40" tumors with morphologic features precisely intermediate between those of SCCL and non-SCCL, including mildly vesicular nuclei, common small nucleoli, and rims of stained cytoplasm. Such tumors are the dividing point between SCCL and non-SCCL and constitute a small fraction of the spectrum from SCCL to non-SCCL. PMID- 2982720 TI - Angiosarcoma associated with germ cell tumors. AB - In two patients with malignant germ cell tumors angiosarcoma developed through two apparently different mechanisms. In one case the angiosarcoma probably developed as a complication of therapeutic radiation, since radiation changes were demonstrated in tissue adjacent to the neoplasm and since the angiosarcoma was not associated with elements of germ cell tumor. The absence of associated germ cell elements does not support the development of the angiosarcoma from a teratoma. In the second case, however, it is likely that the angiosarcoma developed as a result of malignant change within teratomatous foci, since angiosarcomatous elements were intermingled with teratomatous elements and the patient's primary germ cell tumor contained malignant and atypical teratomatous elements as well as prominent vascular proliferation. Malignant change within teratomatous components of germ cell tumors is a phenomenon of increasing importance in this era of effective chemotherapy for germ cell tumors. The development of angiosarcoma as a potential complication of testicular carcinoma has not been reported previously. PMID- 2982721 TI - Oat cell carcinoma of the urinary bladder with ectopic adrenocorticotropic hormone production. AB - A case of primary oat cell carcinoma of the urinary bladder with ectopic adrenocorticotropic hormone (ACTH) secretion in a 55-year-old woman is reported. Cystoscopy revealed a large necrotic tumor in the bladder; the tumor was partially electroresected. At autopsy, a large residual tumor infiltrating the wall of the bladder was found. It had metastasized to the lymph nodes, left ovary, liver, spinal canal, bone marrow, and hypophysis. Bilateral adrenocortical hyperplasia was found. Microscopically, the tumor was identical morphologically to oat cell carcinoma, and the cells contained argyrophilic hormone granules and immunohistochemically demonstrable ACTH. Apparently, oat cell carcinomas with endocrinologic activity have not been reported previously in the urinary bladder. PMID- 2982722 TI - Identification of a case of Y:18 translocation using a Y-specific repetitive DNA probe. AB - We have used a recombinant DNA clone derived from the Y-specific 3.4-kb repeats for in situ chromosome hybridization and Southern blotting analysis to identify a case of de novo Y;18 translocation. The proband has a chromosome complement of 46,XY and a variant chromosome 18 with a Q-bright and C-positive short arm. The father has a normal male karyotype of 46,XY. The mother has a female karyotype of 46,XX and an unusually large Q-bright satellite on one chromosome 22. In situ hybridization with the 3.4-kb probe to the metaphase preparations of family members indicated that the additional Q-bright material in the proband's variant chromosome 18 derived from the Y chromosome of his father, and not from the variant chromosome 22 of his mother. On Southern hybridization, the proband had approximately twice the amount of 3.4-kb repeats per cell as his father. These observations suggest a de novo genetic rearrangement in the proband which probably occurred during the father's spermatogenesis. PMID- 2982723 TI - In situ nick translation of metaphase chromosomes with biotin-labeled d-UTP. AB - Limited nick translation experiments on fixed chromosomes were performed. Sites of preferential DNase-I nicking were made visible by the incorporation of biotin labeled dUTP and subsequent binding of the streptavidin-peroxidase complex. This procedure leads to a banding pattern on the chromosomes which is strongly DNase-I concentration dependent. Along the chromosome arms, regions of enhanced DNase-I sensitivity alternate with regions of lower DNase-I sensitivity. No complete G- or R-type banding pattern was observed. The easily identifiable human Y chromosome was studied more intensively. Compiled data show the heterochromatin of the Y chromosome stained as heavily as the euchromatin. The boundary between the eu- and heterochromatin on the long arm appears to be a site of preferential DNase-I sensitivity. PMID- 2982724 TI - The human thyroglobulin gene: a polymorphic marker localized distal to C-MYC on chromosome 8 band q24. AB - The human thyroglobulin (Tg) gene is localized to chromosome 8 and regionally to band q24 as shown independently by both in situ hybridization techniques and Southern blot analysis of human-rodent somatic cell hybrids. Analysis of hybrids derived from a Burkitt lymphoma, with a translocation breakpoint in the oncogene c-myc, shows that the Tg gene is located distal from c-myc, towards the telomere of the long arm of chromosome 8. The finding of two high frequency restriction fragment length polymorphisms (RFLPs) in the 5' part of the Tg gene results in heterozygosity for at least one marker at chromosome 8, band q24, in 50% of a Caucasian population. These RFLPs cannot only be used for the study of inborn errors in Tg synthesis but also for linkage studies in the telomeric region of chromosome 8q. PMID- 2982725 TI - Detection of a restriction site polymorphism within the human alpha-globin gene complex. AB - A mutant Rsa I restriction endonuclease site of high frequency has been identified in individuals of German, Greek, Italian, and Turkish origin. The mutation was found within the alpha-globin gene complex and is located 0.7 kb 5' to the alpha 2-globin gene. In individuals of central European origin 34 out of 58 chromosomes exhibited the alpha-gene linkage to the presence of the polymorphic site, and thus a preliminary estimate of the gene frequency for this allele would be 0.59. DNA analysis data of individuals derived from Mediterranean populations indicate a distribution of this polymorphic marker in similar frequencies. PMID- 2982726 TI - The human gene encoding insulin-like growth factor I is located on chromosome 12. AB - A cDNA probe corresponding to mRNA encoding human somatomedin-C/insulin-like growth factor I (IGF-I) was used for the chromosomal assignment of the IGF-I gene. Southern-blot hybridization analysis of DNA from human-Chinese hamster somatic cell hybrids showed that the IGF-I gene is located on chromosome 12. Comparison of the chromosomal assignments of the IGF-I gene and two other members of the insulin gene family, with three c-ras oncogenes, reveals a remarkable association of the two gene families. PMID- 2982728 TI - Genetic heterogeneity of delta-aminolevulinate dehydrase and phosphoglycolate phosphatase in north-west Spain. AB - The distribution of delta-aminolevulinate dehydrase and phosphoglycolate phosphatase phenotypes was analyzed in 500 autochthonous individuals from the Galician population (north-west Spain). The gene frequencies for PGP2 and ALADH2 obtained in Galicia have proved to be the lowest of all the European populations so far examined. Comparisons with other world populations were also made. PMID- 2982727 TI - Chromosomal localization and preliminary characterization of the human gene encoding insulin-like growth factor II. AB - A cDNA probe corresponding to mRNA encoding a closely related variant of human insulin-like growth factor II (IGF-II) was used for the chromosomal assignment of the IGF-II gene. Southern blot hybridization analysis of DNA from human-Chinese hamster somatic cell hybrids showed that the human IGF-II gene is located on chromosome 11. Using the same IGF-II variant probe, a cosmid was isolated which contains the human IGF-II gene. Restriction enzyme analysis revealed that the gene encoding IGF-II has a discontinuous structure and contains at least four exons. PMID- 2982729 TI - Inhibition by human T-lymphotropic virus (HTLV-I) of T-lymphocyte mitogenesis: failure of exogenous T-cell growth factor to restore responsiveness to lectin. AB - Various retroviruses, including human T-cell lymphotropic virus (HTLV-I) and avian sarcoma virus (ASV), can prevent coincubated human peripheral lymphocytes from responding efficiently to phytohaemagglutinin. Addition of high concentrations of T-cell growth factor (TCGF) activity usually overcomes this inhibition. However, such restoration of responsiveness does not occur in the case of HTLV-coincubated cells. PMID- 2982730 TI - Preparation of interstitial lung cells by enzymatic digestion of tissue slices: preliminary characterization by morphology and performance in functional assays. AB - A technique is reported here for the quantitative extraction of live cells from the lung interstitium; it involves the incubation of slices of perfused lung in a mixture containing optimal concentrations of collagenase, DNAse, and fetal calf serum, followed by the simultaneous recovery and fractionation of cells released from the tissue matrix on a six-step discontinuous percoll gradient. Yields in the order of 10(8) viable cells per gram of lung were routinely achieved with tissues from rat, mouse and guinea-pig. Preliminary characterization of these cells has been performed in the rat by histological techniques (Giemsa staining, transmission electron microscopy), cytochemistry (acid phosphatase, esterase, peroxidase), by the capacity to bind monoclonal antibodies directed at lymphocyte surface markers, and by a range of functional tests. The cells comprised, on average, 32% macrophages, 44% lymphocytes (T and B cells and large granular lymphocytes), with small numbers of eosinophils, mast cells and epithelial cells. Transmission electron microscopy revealed minimal ultrastructural damage to extracted cells, with such functions as phagocytosis, FcR activity, mitogen responsiveness, antigen presentation, and NK-cell activity, being readily demonstrable. In addition, these activities segregated into defined areas of the six-step density gradient. PMID- 2982731 TI - The mechanism of action of lymphokines. VIII. Lymphokine-enhanced spontaneous hydrogen peroxide production by macrophages. AB - Oil-elicited guinea-pig peritoneal macrophages (MPs) cultured for 2-3 days in medium containing supernatant of concanavalin A-activated lymphocytes (lymphokine, LK) generated large amounts of hydrogen peroxide (H2O2), as detected by the horseradish peroxidase (HRP)-dependent oxidation of phenol red, in the absence of further stimulation. H2O2 production increased with the duration of exposure to LK and was evident at high dilutions of supernatant (1/64). Parallel cultures of MPs in medium or a supernatant of non-activated lymphocytes also increased their H2O2 production during culture but levels at all time intervals were significantly lower than those measured in LK treated cultures. The marked increase in H2O2 production was associated with only a moderate increment in superoxide (O-2) liberation and this was not specific for LK treated cells. Detection of LK-dependent H2O2 production was dependent on ongoing pinocytosis during the assay. This and other arguments suggest that the HRP-phenol red assay, as applied here, detects H2O2 generation occurring at the level of intracellular vesicles and it is concluded that LK elicits H2O2 production that is limited to the intracellular compartment. H2O2 is, apparently, derived by non-enzymatic dismutation of O-2 taking place within the cell; LK treatment of MPs also resulted in a significant reduction in catalase activity. PMID- 2982733 TI - Requirement for cations in the stimulation of C2 synthesis by human monocytes. AB - C2 synthesis by monocytes is stimulated by carbamylcholine acting on nicotinic receptors, phenylephrine acting on alpha 1 adrenergic receptors, and antigen antibody complexes (IC) acting on Fc receptors. Stimulation of C2 synthesis is reversed by agents which block calcium (LaCl2, CoCl2, verapamil, nifedipidine, diltiazem) and sodium channels (tetrodotoxin) and calmodulin antagonists (trifluoperazine and W7). The changes in intracellular cyclic nucleotide levels that follow these receptor-ligand interactions (decreased cAMP, increased cGMP) do not occur in the presence of calcium and sodium channel blockers. These results suggest that the transmembrane signal which is involved in the stimulation of C2 synthesis is the entry of sodium and calcium ions. Whether this influx occurs by separate channels or a common channel has not been determined. The intracellular events involved in the stimulation of C2 synthesis appear to be calmodulin-dependent. PMID- 2982732 TI - Suppression of delayed-type hypersensitivity mediated by macrophage-like cells in mice with experimental liver injury. AB - The intensity of picryl chloride-induced delayed-type hypersensitivity (PCl-DTH) was significantly lowered in mice with experimental liver injury induced by the injection of carbon tetrachloride (CCl4) or 1-naphthylisothiocyanate (ANIT). Adherent spleen cells prepared from mice with liver injury suppressed the PCl-DTH in normal syngeneic mice by adoptive transfer at the time of immunization with picryl chloride (PCl). Cianidanol, administered orally to recipient mice immediately after the transfer of adherent spleen cells, caused total prevention of the PCl-DTH from suppression by the transferred adherent spleen cells. In in vitro studies, the adherent spleen cells from both CCl4-treated and ANIT-treated mice produced higher amounts of prostaglandin E2 (PGE2) than those from normal mice. The addition of cianidanol at the beginning of incubation caused an inhibition of PGE2 production of the adherent spleen cells. Furthermore, the spleen cell suspensions from CCl4-treated mice gave a marked increase in generation of superoxide anions (O2-), also inhibited by the addition of cianidanol. Gathered results support the adherent spleen cell as being the cell causing the suppression of DTH in mice with experimental liver injury; the important role of PGE2 and O2- in the suppression of DTH mediated by the adherent spleen cells was demonstrated. Cianidanol eliminated the suppression of DTH, owing principally to the inhibitory effect on the production of PGE2 and O2- from the adherent spleen cells. PMID- 2982734 TI - Transferrin receptor expression by stimulated cells in mixed lymphocyte culture. AB - Transferrin receptor (TRFr) expression by cells in mixed lymphocyte culture increases steadily for the first 5 days, but then reaches a plateau. By the sixth day in culture, about 20% of viable cells express TRFr in two-way mixed lymphocyte reactions. This subpopulation of TRFr-positive cells represents the proliferating population; it is heterogeneous, containing T-cell blasts and smaller cells which are a mixture of T and non-T cells. A small group of non-T cells have phenotypic similarity to natural killer (NK) cells. T cells appear to divide earlier in the course of the response than non-T cells. The biphasic nature of this response and the slower non-T reactivity may be due to a secondary stimulation of non-T cells by factors released from activated T cells (such as interleukin-2). PMID- 2982735 TI - Binding site and subclass specificity of the herpes simplex virus type 1-induced Fc receptor. AB - Immunoglobulin Fc-binding activity was detected by indirect immunofluorescence employing fluorochrome conjugated F(ab')2 antibody fragments on acetone-fixed cell cultures infected with herpes simplex virus type 1 (HSV-1). Using this method the Fc receptor-like activity seemed to be restricted to the IgG class of human immunoglobulins. While IgG1, IgG2, and IgG4 myeloma proteins bind to this putative Fc gamma receptor at a concentration of 0.002 mg/ml, IgG3 myeloma proteins were without activity at 0.1 mg/ml. The binding activity was associated with the Fc fragments of IgG, while the pFc' fragments of IgG appeared to be unable to bind in this assay system. The reactivity and specificity of the HSV-1 Fc receptor was independent of both the type of tissue culture cells used and the strain of HSV-1 inducing the Fc receptor-like activity. The HSV-1-induced Fc receptor has a similar specificity for human immunoglobulin class and subclasses as staphylococcal Protein A. However, these two Fc receptors exhibit at least one striking difference. The IgG3 G3m(st) protein which binds to Protein A does not bind to HSV-1-induced Fc receptor. A possible reaction site for the HSV-1 Fc receptor on IgG could be at or near Asp 276. PMID- 2982737 TI - Oxidation of microbial iron-sulfur centers by the myeloperoxidase-H2O2-halide antimicrobial system. AB - Myeloperoxidase, H2O2, and a halide (chloride, bromide, or iodide) form a potent microbicidal system that contributes to the antimicrobial activity of neutrophils. The mechanism of toxicity is not completely understood. Powerful oxidants are formed that presumably attack the microbe at a variety of sites. Among the consequences of this attack is the release of a large proportion of 59Fe of prelabeled organisms. We report here that the myeloperoxidase-H2O2-halide system oxidizes the iron-sulfur centers of model compounds (spinach ferredoxin) and intact microorganisms (Escherichia coli) with the loss of labile sulfide. The oxidation of the iron-sulfur centers of ferredoxin was measured by the fall in absorbance at 420 nm (bleaching) and by the loss of 5,5'-dithiobis-(2 nitrobenzoic acid) reducing activity. The latter compound is a sulfhydryl reagent that is reduced by ferredoxin labile sulfide during denaturation. The oxidation of E. coli iron-sulfur centers by the peroxidase system was determined by the loss of labile sulfide content, as measured by the release of H2S by acid and its reaction with zinc acetate to form ZnS. The halides were effective as components of the peroxidase system in the order I greater than Br greater than Cl. The oxidation of E. coli iron-sulfur centers by the peroxidase system was rapid and preceded the loss of viability. Gentamicin, at a concentration which produced a loss of viability comparable to that of the peroxidase system, did not cause a loss of labile sulfide from E. coli, suggesting that labile sulfide loss is not a nonspecific reflection of the loss of viability, but a direct consequence of the action of the myeloperoxidase system. The oxidation of iron-sulfur centers in microorganisms by the myeloperoxidase-H2O2-halide system may contribute to the death of the organism. PMID- 2982736 TI - Resistance to adenovirus infection after administration of Bordetella pertussis vaccine in mice. AB - Treatment of mice with Bordetella pertussis vaccine rendered mice resistant to mouse adenovirus infection. The resistant state took at least 5 days to develop, and susceptibility returned to a portion of the test population 35 days after treatment. Transient resistance developed in congenitally athymic mice also. Treatment with a dose of 25 micrograms (dry weight) of B. pertussis vaccine protected approximately 50% of the test population. Vaccines prepared from several different strains of B. pertussis were capable of inducing resistance, and the induction of resistance was not dependent on the mouse strain used for testing. Cross-reacting antibodies capable of neutralizing the virus or protecting against a challenging infection were not induced by treatment with B. pertussis vaccine. PMID- 2982739 TI - Naturally occurring antibodies in human sera that react with the iron-regulated outer membrane proteins of Escherichia coli. AB - Sera from normal healthy human adults and infants, as well as sera from mice, rabbits, and guinea pigs, were examined by immunoblotting for naturally occurring antibodies reacting with outer membrane proteins of two Escherichia coli strains, O111 and O18. Some individuals had antibodies reacting very strongly with the iron-regulated outer membrane proteins, including the ferric-enterochelin receptor protein (Mr, 81,000), as well as with ompA. However, sera from infants contained predominantly antibodies to ompA; antibodies recognizing the iron regulated outer membrane proteins were either absent or barely detectable. In human serum the antibodies were mainly of the immunoglobulin G class. No serotype specific antibodies to the lipopolysaccharide of E. coli O111 or O18 were found in the sera tested. PMID- 2982738 TI - Cloning of extracellular DNase and construction of a DNase-negative strain of Vibrio cholerae. AB - The structural gene xds for extracellular DNase of Vibrio cholerae was cloned and inactivated by insertion of the transposon Tn5. The inactivated gene was introduced into the chromosome of V. cholerae by recombination to construct an extracellular DNase-negative strain. Tn5-mediated transposon-facilitated recombination was used to establish the position of xds between the pro-1 and ile 201 markers on the genetic map of V. cholerae. The extracellular DNase-negative strain described here should be useful for investigating the role of the xds encoded DNase in the physiology of V. cholerae and its plasmids as well as for characterizing other DNases in this organism. PMID- 2982740 TI - Possible relationship of a 36-megadalton Salmonella enteritidis plasmid to virulence in mice. AB - All of the Salmonella enteritidis strains isolated from diseased animals (61 strains) and from beef (2 strains) in Japan and in West Germany (1 strain), except for 2 strains isolated from ducks, harbored either a 36-megadalton (Md) plasmid alone or in combination with several other plasmids of different sizes. It is likely that these 36-Md plasmids from various S. enteritidis strains were derived from the same origin because their plasmid DNAs showed the same cleavage patterns obtained with EcoRI, HindIII, and BamHI. We also suggested that this plasmid is native to S. enteritidis. Tests carried out on two strains isolated from ducks which naturally lacked this plasmid and one strain whose plasmid was artificially cured showed that the strains without the 36-Md plasmid showed less virulence compared to a wild-type strain harboring the 36-Md plasmid, suggesting that this 36-Md plasmid might be associated with virulence for mice. PMID- 2982741 TI - Generation of oxygen species and virulence of Listeria monocytogenes. AB - Listeria monocytogenes strains differing in virulence were examined for production of oxygen species. Virulent strains A4413 and JH-H exhibited maximal generation of hydrogen peroxide and superoxide, whereas the avirulent strain 9037 7R was least active. The virulence was confirmed by the in vitro assessment of the intracellular fate of the microorganisms in resident macrophages. PMID- 2982742 TI - Reduced adenylate cyclase responsiveness to histamine in lymphocyte membranes of allergic asthmatic patients after allergen challenge. AB - The adenylate cyclase response to histamine was measured in lymphocyte membranes of allergic asthmatic patients and healthy control subjects, just before and 24 h after inhalation challenge with house dust mite allergen. In the non-acute phase before allergen challenge, the histamine response in the membranes of the patients was not significantly different from that in membranes of the control subjects. After the house dust mite-induced asthmatic reaction, however, the adenylate cyclase response to histamine in the membranes of the patients was significantly reduced by 47%, whereas allergen inhalation had no effect on the histamine response of the control membranes. The results suggest that allergic asthmatic reactions may cause reduced lymphocyte adenylate cyclase responsiveness to histamine. A similar mechanism in the airways might contribute to allergen induced enhanced airway reactivity to histamine. PMID- 2982743 TI - Bicarbonate solutions for peritoneal dialysis: a reality. PMID- 2982744 TI - Prevalence of human papillomavirus type 16 DNA in cervical carcinoma samples in East Anglia. AB - A substantial increase in the incidence of severely dysplastic cervical lesions (CIN 3) has been observed during the period 1975-1982 in the East Anglian region of England. Since patients with severe dysplasia have an enhanced risk of developing cervical carcinoma, it seems possible that a substantial increase in the rate of cervical carcinoma is likely to occur in the near future. Evidence of a relationship between human papillomavirus (HPV) infection and cervical carcinoma has accumulated recently. We have studied the incidence of HPV16 DNA in cervical tissue samples from patients with cervical carcinoma, severe dysplasias and normal controls. Five out of 11 invasive squamous carcinomas of the cervix, 3/4 dysplasias and 0/12 normal samples were positive in Southern blot assays for HPV16 DNA. Some of the tissue samples had as many as 500 copies of HPV16 DNA per cell. The amount of HPV16 DNA present correlated with the aggressiveness of tumour growth. PMID- 2982745 TI - Capacity of B-lymphocytic lines of diverse tumor origin to produce and respond to B-cell growth factors: a progression model for B-cell lymphomagenesis. AB - Human cell lines established from cases of acute lymphoblastic leukemia, lymphosarcoma, Burkitt's lymphoma and multiple myeloma and representing stages of B-lymphocyte development ranging from pre-B through to plasma cells, were assessed for their ability to produce and respond to B-cell growth factors (BCGF). All B-cell lines studied were found to be constitutive producers of a growth activity which assisted the S-phase entry of normal activated B-cells and provided growth support for lymphoblastoid cells transformed by Epstein-Barr virus. Furthermore, all lines responded by enhanced proliferation to supernatants from a BCGF-producing T-cell hybridoma. Not all lines, however, displayed autostimulation to their own supernatants and no tumor B-cell line appeared totally dependent on soluble factors for its growth. Non-tumorigenic B-cell lines, by contrast, revealed a strict dependency on homologous growth factor for their continued proliferation in suspension culture. The findings support a progression model of lymphomagenesis based upon the utilization, production and, ultimately, emancipation from growth-promoting soluble factors. PMID- 2982746 TI - Identification of Marek's disease virus-specific antigens in Marek's disease lymphoblastoid cell lines using monoclonal antibody against virus-specific phosphorylated polypeptides. AB - For identification of the antigens specific to Marek's disease virus (MDV) in virus-non-producer lymphoblastoid cell lines established from a tumor of Marek's disease (MD), hybridomas producing monoclonal antibodies (MAbs) against the antigens were isolated. Immunogens for preparation of the hybridomas were purified from the lysate of an MD-lymphoblastoid cell line, MSB1, by affinity chromatography coupled with chicken anti-MDV serum immunoglobulin G. Three of the MAbs obtained, MB1, MB2 and MB3, were specific to MDV by immunofluorescence test. An immunofluorescence test using MB2 antibody showed that immunofluorescence positive cells in non-producer MD-lymphoblastoid line cells became detectable when the culture temperature was shifted from 41 degrees C to 33 degrees C or when treatment with 5-iodo-2-deoxyuridine (IUdR) was performed, indicating that the antigen reactive with MB2 antibody is an MDV-specific early antigen. This temperature shift or IUdR-treatment did not induce other MDV-specific antigens, such as late gene products of MDV, detected with MAbs. MB1 and MB2 antibodies immunoprecipitated 4 MDV-specific phosphorylated polypeptides with molecular weights (MWs) of 43,000 (43kd), 39kd, 36kd and 24kd from chick embryo fibroblasts productively infected with virulent MDV. In the place of 43kd, phosphorylated 44kd polypeptide was precipitated from avirulent MDV-infected fibroblasts. However, MB3 antibody did not precipitate any MDV-specific polypeptides from infected fibroblasts. These results suggest that the phosphorylated polypeptides are MDV-specific polypeptides predominantly expressed in non-producer MD lymphoblastoid cell lines. PMID- 2982747 TI - Seroepidemiology of human T-lymphotropic retrovirus type I (HTLV-I) in residents of Niigata Prefecture, Japan. Comparative studies by indirect immunofluorescence microscopy and enzyme-linked immunosorbent assay. AB - A large sample of carriers of human T-lymphotropic retrovirus type I (HTLV-I) in Niigata Prefecture was examined for the detection of natural antibodies to HTLV-I related antigens in sera using both indirect immunofluorescence microscopy (IFM) and enzyme-linked immunosorbent assay (ELISA). The present findings are based on multiple surveys, using each assay technique at least twice. Although Niigata Prefecture has been considered a non-endemic region for HTLV-I, Sado Island has been proven by this study to be a relatively endemic pocket within this non endemic area. Seropositivity was highest in residents of Sado Island; 97/1, 117 (8.7%) by IFM and 33/1,061 (3.1%) by ELISA; followed by Niigata City, 18/650 (2.8%) by IFM and 16/638 (2.5%) by ELISA; and lowest in the remaining areas, 57/2,631 (2.2%) by IFM and 20/2,551 (0.8%) by ELISA. Seropositivity was demonstrated in 172/4,398 (3.9%) by IFM and 69/4,250 (1.6%) by ELISA in Niigata Prefecture taken as a whole. In general, the incidence of seropositive residents increased gradually with age. The sex difference was not significant. The serum samples tested were categorized into 4 groups; (1) IFM and ELISA both positive, (2) IFM positive but ELISA negative, (3) IFM negative but ELISA positive, and (4) IFM and ELISA both negative. By absorption tests, IFM and ELISA seemed to recognize different specific antibodies in sera; IFM recognized antibodies to HTLV-I-related cellular antigens in addition to HTLV-I viral antigens, but ELISA recognized antibodies to HTLV-I viral antigens alone. Thus, IFM detected a broader spectrum of antigens, resulting in recognition of more positive sera than those detected by ELISA. PMID- 2982749 TI - Characterization of a second Epstein-Barr virus-determined nuclear antigen associated with the BamHI WYH region of EBV DNA. AB - The Epstein-Barr virus-determined nuclear antigen (EBNA) is the only known virally-determined component that is regularly associated with EBV-transformed cells. A main component of EBNA, herein designated EBNA-1, has been conclusively localized to the BamHI K fragment of the viral genome. EBNA-1 is present in all EBV-carrying cell lines so far studied. Our current study deals with a second component. We have found that the EBNA reaction detected by anti-complement immunofluorescence (ACIF) in Burkitt lymphoma lines Daudi, Jijoye, and P3HR-1 could be completely removed by preabsorption of sera with any one of these 3 lines, when tested against any other of them. The same absorbed sera still gave a brilliant nuclear staining against other EBV-carrying lines, e.g. Raji or B95-8. The 3 lines in the first category carry EBV genomes that have deletions in the BamHI WYH region of the EBV genome. This region is intact in the second group of lines. This result is interpreted as showing the existence of 2 different ACIF stainable EBV-determined nuclear antigens, one of which is associated with the BamHI WYH region. We designate this antigen as EBNA-2. We found that the two different EBNAs are different with regard to their association with metaphase chromosomes. In lines positive for both EBNA subtypes, metaphase chromosomes gave brilliant EBNA-1 staining, but could not be stained for EBNA-2, indicating differences in chromatin association of the two EBNAs. An 86 kd polypeptide was identified by immunoblotting of DNA-binding proteins from EBV-transformed lymphoid cell lines. EBV-specificity of the polypeptide was demonstrated by the presence of antibodies against this polypeptide in antisera from a population of EBV-seropositive donors, but not from seronegative donors, by the presence of the polypeptide itself in EBV-carrying but not in EBV-negative cell lines and by the appearance of antibodies against this polypeptide during the course of infectious mononucleosis (IM). The polypeptide was absent from the EBV-carrying P3HR-1, Daudi and Jijoye cell lines, which suggested that it may be encoded by the BamHI WYH region that is deleted from the viral substrains carried by these lines. PMID- 2982750 TI - Converting-enzyme inhibition for severe chronic heart failure: views from a skeptic. PMID- 2982748 TI - Effect of cyclosporin-A (CsA) on the ability of T lymphocyte subsets to inhibit the proliferation of autologous EBV-transformed B cells. AB - We have analyzed the effect of Cyclosporin-A (CsA) on the in vitro suppression of Epstein-Barr virus (EBV)-induced B-cell proliferations separated on the basis of cell density. CsA abolished the growth-inhibitory capacity of high-density T cells but influenced only marginally the activity of low-density lymphocytes; this suggested that different mechanisms mediate suppression in the two subsets. Stimulation with irradiated EBV-transformed cells had a different impact on the activity of low- and high-density lymphocytes. Proliferative and cytotoxic responses were inversely correlated, i.e. high-density cells proliferated but exerted low cytotoxicity, while the lytic activity of the low-density subset was stronger in the absence of significant cell proliferation. Proliferation and generation of cytotoxicity were abrogated by CsA in both subsets. The activities could be restored by addition of exogenous IL-2, suggesting that the drug may interfere with the cascade of lymphokine--cell interactions which leads to activation of immune responses. From the analysis of athe CsA effects on the two subsets it seems that the high-density one contains specific memory T cells which are activated and proliferate upon encounter with EBV-infected cells. On the other hand, low-density lymphocytes are induced to release soluble factors with antiproliferative activity. The secretory function was resistant to the suppressive effect of CsA. PMID- 2982751 TI - Alkaline phosphatase promotes radioprotection and accumulation of WR-1065 in V79 171 cells incubated in medium containing WR-2721. AB - Addition of alkaline phosphatase and WR-2721 to culture medium containing V79-171 cells leads to production of WR-1065 and its disulphide forms in the medium, to cellular accumulation of WR-1065, and to radioprotection which correlates with cellular WR-1065 level. PMID- 2982752 TI - Update on AIDS. PMID- 2982753 TI - The phakomatoses. PMID- 2982754 TI - Drug responses of adenylate cyclase in iris-ciliary body determined by adenine labelling. AB - The intracellular adenine nucleotide pool of rabbit iris-ciliary body was labelled by uptake of 3H-adenine in vitro. A variety of agents were tested for their ability to stimulate or inhibit the incorporation of radioactivity into cyclic AMP formed from ATP labelled with 3H-adenine. Isoproterenol, vasoactive intestinal peptide, forskolin, and prostaglandin E2 stimulated incorporation of label 3-10-fold in 15-20 min compared with paired tissues not treated with hormone, whereas histamine, serotonin, substance P, and bradykinin were inactive. Clonidine, alpha-methylnorepinephrine, and dopamine decreased the rate of incorporation of label into the cyclic-AMP pool in tissues that showed high spontaneous basal rates. In low-basal tissues these drugs were inactive by themselves but clonidine and alpha-methylnorepinephrine blocked the stimulation effected by isoproterenol. The findings indicate that several receptor-coupled adenylate cyclase systems are present in ICB and that dual adrenergic control of adenylate cyclase through positive and negative coupling of adrenergic receptors probably occurs. The negatively coupled adrenergic receptors appear to be similar to the alpha 2-subclass of adrenergic receptor described in other tissues. These observations suggest a role for the large number of alpha 2-adrenergic-binding sites found in albino rabbit iris-ciliary body by ligand binding assays. PMID- 2982755 TI - Interactions of Junin and Tacaribe viruses during mixed infections. AB - The interaction between Junin virus (JV) and Tacaribe virus (TACV) during mixed infections of RK13 cells was examined. The effects of a prior infection with JV upon TACV replication depended on the time between the two inoculations. Simultaneous infection of RK13 cells with TACV and JV did not alter the plaquing efficiency of TACV; but if there was a 1- to 24-hour delay between JV preinfection and TACV superinfection, a variable increase of TACV replication was observed. The enhancement of TACV replication by preinfection with JV was dependent on several factors, such as the MOI of both viruses and the integrity of the JV genome. This effect was also highly specific, as the plaquing efficiencies of the arenavirus Pichinde and the unrelated vesicular stomatitis virus were not affected by preinfection with JV at any multiplicity assayed. The majority of the progeny formed in cells superinfected with TACV 1 or 24 h after JV infection was partially neutralized by antisera to both viruses. This suggested that phenotypic mixing, with JV or TACV genomes enclosed within an envelope containing TACV and JV glycoprotein, had occurred. PMID- 2982756 TI - Characterization of adenovirus 15/H9 intermediate strains. AB - 17 adenovirus strains from various parts of the United States and Europe were characterized by immunological and biochemical methods. They were antigenically related to adenovirus 15 (Ad15) in neutralization tests. In hemagglutination inhibition tests, a relationship was seen to Ad9, but not to Ad15. These strains were named Ad15/H9 intermediate strains. The relatedness of prototypes Ad15 and Ad29 in neutralization tests and of Ad9 and Ad8 in hemagglutination-inhibition tests was also found in the strains studied. The DNAs of these strains were analyzed by use of restriction endonucleases. After cleavage with the enzymes BamHI, BglII, BstEII, HindIII, and SmaI, fragment patterns of Ad15/H9 strains were found to be different from the prototypes and were more closely related to Ad9 than to Ad15. Furthermore, the 17 strains could be classified into six different genome types, a-f. PMID- 2982757 TI - Characterization of the DNA of canine adenovirus by restriction enzyme analysis. AB - The DNA of canine adenovirus (canAV 1, strain Behring H.c.c. 269) was characterized in detail. The molecular weight of canAV DNA was (20.1 +/- 0.1) x 10(6) daltons as determined by contour length measurements, and 19.9 x 10(6) daltons as determined by restriction enzyme analysis. A terminal protein was found to be covalently linked to the 5' ends of canAV DNA. Physical maps of viral DNA were constructed for BamHI, ClaI, SacI, SalI, and SmaI enzymes. Three international strains of canAV [American Type Culture Collection (ATCC) catalogue No. VR-800, VR-293, and VR-133] were analyzed under the same conditions. CanAV 1 (ATCC VR-133 and VR-293) had DNA cleavage patterns identical to the H.c.c. 269 strain, whereas the DNA cleavage pattern of canAV 2 (ATCC VR-800) was clearly different. PMID- 2982758 TI - RSV provirus with same flanking sequences is found on different size classes of Chinese hamster chromosomes. AB - Rous sarcoma virus(RSV)-transformed Chinese hamster fibroblasts, containing approximately ten copies of the DNA domain comprising a single provirus and its flanking cellular sequences, were arrested in metaphase, and the chromosomes were fractionated by size in a sucrose gradient. The resolution of polymorphic ribosomal genes, the dihydrofolate reductase gene, and the c-src gene demonstrated that the gradient can distinguish between small, medium, and large chromosomes. The same DNA domain carrying the RSV provirus was found to be associated with chromosomes of all three size classes. Polymorphic copies of the domain in small and large chromosomes could be distinguished from those in medium sized chromosomes because of the polymorphism in the XhoI and EcoRI sites on 5' and 3' adjacent cellular sequences, respectively. The presence of the same provirus domain on different chromosomes, together with the karyological data showing trisomies in the same chromosome size classes, suggest that the provirus domain, possibly the entire replicon, was duplicated and transferred to different nonhomologous chromosomes, and the transfer was followed by duplication of the target chromosomes. The possibility that proviral long terminal repeats might be involved in replicon transfer is discussed. PMID- 2982759 TI - Antiviral activity of CP-20,961 against herpes simplex viruses in vitro. AB - The antiherpesvirus activity of CP-20,961 [N,N-dioctadecyl-N',N'-bis (2 hydroxyethyl) propanediamine, or Avridine] was investigated in cultured guinea pig embryo (GPE) cells. Plaque formation of herpes simplex virus type 1 (HSV1) and type 2 (HSV2) was inhibited, but vesicular stomatitis virus replication was not inhibited, in GPE cells treated with CP-20,961 before infection. The ID50 concentration of CP-20,961 for HSV was about 50 micrograms/ml for 3-4 days of pretreatment. After virus adsorption and penetration, the same concentration of CP-20,961 had no effect on HSV plaque formation. The compound showed no detergent like properties nor did it elicit any detectable interferon activity. Thus, the anti-HSV activity of CP-20,961 appeared to be associated with blocking the adsorption or penetration of the virus or both. PMID- 2982760 TI - Effect of n-butyrate on transformation of human lymphocytes by Epstein-Barr virus. AB - 3 mM n-butyrate, which effectively induces synthesis of Epstein-Barr virus (EBV) antigens in the producer P3HR-1 cell line, inhibited the transformation of umbilical cord blood lymphocytes by EBV. The drug inhibited both EBNA synthesis and stimulation of host cell DNA synthesis by strain B95-8. This effect was less pronounced at lower concentrations, but even at 0.1 mM (which did not affect DNA synthesis in EBNA and stimulation of host cell DNA synthesis in EBV-infected lymphocytes were delayed. PMID- 2982761 TI - Alkaline deoxyribonuclease induced with diterpene ester TPA and n-butyrate in Epstein-Barr virus genome-carrying Raji cells. AB - The combination of 12-O-tetradecanoylphorbol-13-acetate and n-butyrate induced Epstein-Barr virus (EBV)-associated alkaline DNase in Raji cells. The anti-early antigen-positive sera neutralized EBV-associated DNase activity. The enzyme activity was eluted at about 0.20 and 0.24 M KCl from DEAE-cellulose and phosphocellulose columns, respectively. The partially purified enzyme was sensitive to (NH4)2SO4 and spermine. Immunological and biochemical properties of EBV-associated DNase induced in Raji cells were similar to those of the enzyme induced in P3HR-1 cells. PMID- 2982762 TI - Infection of pregnant guinea pigs with attenuated Junin virus strains. AB - The effect of the attenuated XJC13 and XJ0 strains of Junin virus (JV) was studied in guinea pigs infected before and during pregnancy. The 58% mortality rate in animals infected during gestation and the 16.7% mortality rate in chronically infected animals were attributed to a viral effect. An abortion rate of 33% occurred in animals infected before the 7th week of gestation. Regardless of the time of infection, JV was isolated from central nervous system tissue, placentas, and fetuses of animals killed just before parturition, even when circulating neutralizing antibodies were present. Results confirmed that transplacental infection is a regular event and showed that guinea pigs are more susceptible to attenuated JV strains during pregnancy, most probably due to immunosuppression, hormonal changes, or both. PMID- 2982763 TI - Intravenous acyclovir for herpesvirus in immunocompromised patients. AB - Severe herpesvirus infections in 18 immunocompromised patients (25 episodes) were treated with i.v. acyclovir. Six patients had 13 episodes of mucocutaneous herpes simplex infections, eight children had varicella, three patients had generalized zoster, and one patient had cytomegalovirus pneumonia. No patient died from infections. In 18 episodes treatment with acyclovir produced a beneficial effect, in 5 episodes acyclovir was probably beneficial, and in 2 patients the effect could not be evaluated. No serious side effects could be definitely attributed to acyclovir administration. These data support the previous experience that i.v. acyclovir may be useful in the treatment of herpesvirus infections in immunocompromised patients. PMID- 2982764 TI - Vitamin D-dependent rickets types I and II. Diagnosis and response to therapy. AB - The diagnostic value of measuring serum vitamin D metabolites is demonstrated in the present study in which two patients with vitamin D-dependent rickets (VDDR) Types I and II are reported. The patient with sporadic VDDR Type I was severely disabled and unable to walk. Her serum 1,25(OH)2D level was low (32 pg/ml, normal 30 to 60) and she responded dramatically to 1 microgram of 1 alpha-hydroxyvitamin D3 daily. The VDDR Type II patient had an autosomal recessive inheritance and total alopecia. His serum 1,25(OH)2D level was greater than 500 pg/ml, compatible with end-organ refractoriness to 1,25(OH)2D. He did not respond to daily doses of 1 microgram 1 alpha-hydroxyvitamin D3. These cases demonstrate the striking difference in 1,25(OH)2D levels and therapeutic response to 1,25(OH)2D in these two conditions. PMID- 2982765 TI - Endorphin levels in headache syndromes. PMID- 2982766 TI - Neutron skyshine measurements at Fermilab. AB - Neutron skyshine has been a significant source of environmental radiation exposure at many high-energy proton accelerators. A particularly troublesome source of skyshine neutrons has existed at Fermilab during operation of the 400 GeV high-energy physics program. This paper reports on several measurements of this source made with a DePangher precision long counter at large distances. The spatial distribution of the neutron skyshine can approximately be described as an inverse square law dependence multiplied by an exponential with an approximate attenuation length of 1200 +/- 300 m. The absolute magnitude of the distributions can be matched directly to the conventionally measured absorbed dose distribution near the source. PMID- 2982767 TI - Rous sarcoma virus SRC gene expression on the growth of quail embryo skin fibroblasts and the establishment of permanent cell lines. AB - Permanent cell lines of Quail embryo fibroblasts appear in cultures of cells infected with a wild type strain of Rous sarcoma virus (SR-RSV) or with its temperature sensitive transformation mutants (ts-T) (NYts68 and PA101) following a three step process. In step one, infected cells grow twice as fast as the control. The second step consists of a crisis during which the cell population is stationary for four to five weeks. Towards the fourth week several foci of cell growth are observed in the flasks. Respreading of the content of these flasks yields permanent lines. This constitutes the third step of the population evolution. In step one the growth rate of the infected cells is the same irrespective of the incubation temperature (36 degrees C or 41 degrees C) whereas the level of the pp60v-src activity is considerably depressed at 41 degrees C for NYts68 and PA101. Foci do not appear at restrictive temperature in the ts infected population and permanent lines are not recovered under that condition. These lines grow ony at 36 degrees C. It can be shown that the virus which they produce is not modified with respect to the temperature sensitivity of the src gene expression since newly infected fibroblasts grow equally well in step one at both 36 degrees C and 41 degrees C, and stop after the same number of generations. This finding suggests that the events which, during the crisis period, lead to the establishment of permanent lines, take place at the cellular level but depend on the activity of the pp60v-src protein for their occurrence or their expression. PMID- 2982768 TI - Pathogenesis and virus content of lymphomas induced by pure ecotropic Graffi murine leukemia virus. AB - Tumors induced by wild type Graffi murine leukemia virus (Gi-MuLV) contained high titers of MuLV consisting of a predominant ecotropic (e)-MuLV and a scarcer titer recombinant (RM) MuLV component. Each of these was purified by biological cloning and examined for its envelope properties and leukemogenicity. Both the e- and the RM-MuLV's were single isolates and unique in terms of their neutralization profiles and peptide maps. The cloned e-Gi-MuLV was highly leukemogenic in C57Bl mice, inducing a very rapid lethal thymic lymphoma but no myeloid leukemia. e-Gi MuLV also accelerated thymic lymphoma in AKR mice. The purified RM-MuLV did not induce any tumors. Infectious cell center (ICC) experiments of organs of mice inoculated with e-Gi-MuLV showed that virus replicated very rapidly and reached maximal titers in about one week in C57Bl mice. There was a highly preferential replication in the thymus of the animal so that this e-Gi-MuLV can be considered as thymotropic. Within two weeks after infection of mice, infected cells of the thymus also began to release low levels of a non-ecotropic MuLV. The rapid induction of lymphoma is compared to that induced by other e-MuLV's and their RM MuLV's, and to the natural AKR-MuLV-associated disease. These findings are discussed in the context of prevailing theories on envelope gene rearrangements in the virus and in the proviral sequences in resulting tumors. PMID- 2982769 TI - The formation of radiation-induced DNA breaks: the ratio of double-strand breaks to single-strand breaks. AB - Ionizing radiation causes the formation of strand breaks in cellular DNA, as well as other types of lesions in the chromatin of cells. Some of the earliest investigations of the molecular basis of radiation-induced damage and the implications of enzymatic repair were done by Dr. H. S. Kaplan. The induction frequency of DNA double-strand breaks is of special importance, and it is of interest to know the relative proportions of single-strand and double-strand breaks. This ratio changes noticeably with the radiation quality (ionization density). Because it is difficult to assay for DNA lesions in the large mammalian genome, we have developed a method of assaying for DNA double-strand breaks in the supercoiled nucleosome-complexed Simian virus 40 (SV40) genome, irradiated intracellularly. In this communication we present our measurements of the DNA double-strand breaks (DSBs) to single-strand breaks (SSBs) ratio obtained from the intracellularly irradiated SV40 genome. After cobalt gamma ray and X ray irradiations, this ratio is about 1/10. Our methods and results are compared with pertinent data in the literature. If the DSBs/SSBs ratio of 1/10 for cellular chromatin is correct, a substantial number of DNA double-strand breaks are formed in a mammalian cell after moderate doses (1 Gy) of radiation. The implications of different types of DNA double-strand breaks are discussed. PMID- 2982770 TI - Long-term results of local excision and radiotherapy in pleomorphic adenoma of the parotid. AB - Three hundred and eleven patients with primary pleomorphic salivary adenoma of the parotid gland seen between 1950 and 1971 are reviewed. All were managed by a combination of surgery, usually local excision, and irradiation. The minimum follow-up is 10 years. Recurrence rates at 0-5 years and 5-10 years were 1 and 1.5%, respectively, and all recurrences in this period were histologically or clinically benign. This recurrence rate is similar to those reported after superficial parotidectomy. After 10 years, however, the recurrence rate rose, becoming maximal at 15-20 years (4%) and yielding a cumulative risk of recurrence of 8.0% at 20 years. The late recurrences were predominantly malignant tumors. One tumor was probably radiation-induced; the remainder were compatible with spontaneous malignant transformation of benign pleomorphic adenoma, although radiation may have increased the incidence. In the absence of a comparable purely surgical series, this cannot be determined. The recommendation is made that the management of these tumors should be primarily surgical with irradiation reserved for patients presenting surgical difficulties. In addition, prolonged follow-up is necessary since there is a significant incidence of late recurrence and such recurrence may be more likely to be malignant regardless of the primary form of management. PMID- 2982771 TI - Sequential hemibody and local irradiation with combination chemotherapy for small cell lung carcinoma: a preliminary analysis. AB - Sequential hemibody irradiation (SHB) was integrated with combination chemotherapy and local irradiation (LRT) in the induction and consolidation phases of a therapeutic protocol for small cell lung carcinoma (SCLC). Forty-one previously untreated patients were entered into this program. Among 38 evaluable patients (20 with limited disease [LD] and 18 with extensive disease [ED], the overall response rate was 63% (90% in LD and 33% in ED patients). The estimated overall survival is 8.1 months. The major toxicity has been myelosuppression- especially thrombocytopenia. The frequency of previously described "acute radiation syndromes" and radiation pneumonitis associated with hemibody irradiation have been substantially decreased at the current dosage with premedication and shielding techniques. The integration of SHB as a systemic therapy with combination chemotherapy and LRT is a feasible program for sequential administration of non-cross-resistant agents in SCLC and may be beneficial in patients with limited disease. PMID- 2982772 TI - The evaluation of radiotherapy after incomplete surgery in patients with carcinoma of the maxillary sinus. AB - The retrospective analysis of 57 patients with cancer of the maxillary antrum irradiated after incomplete surgery was performed. The majority of patients had very advanced disease (54% T4 tumors). In 18 patients partial resection of maxillary antrum was performed: 39 patients underwent total maxillectomy. In 35 patients macroscopic residual tumor (MRT) was present after surgery. All patients were irradiated postoperatively with 60Co teletherapy and received a dose of 60 Gy in 20-30 fractions over 4-6 weeks. Five year symptom-free survival in the whole group was 35%. Significantly better survival was found in patients with T2 tumors in comparison with patients with T3 and T4 tumors, in patients with infrastructural localization in comparison with patients with suprastructural localization and in patients with microscopic residual tumor in comparison with MRT patients. An analysis of pattern of relapses indicates that histology should be regarded as an important factor of management. In keratinizing squamous cell cancer, local control remains the main problem. In patients with nonkeratinizing squamous cell cancer, both local and regional control is important and elective irradiation of neck nodes may be of value. In patients with undifferentiated cancer, distant metastases appear to have the greatest impact on survival. PMID- 2982773 TI - Protection of mouse lung by WR-2721 after fractionated doses of irradiation. AB - The radioprotective effect of WR-2721 on mouse lung has been studied after single doses, 4 or 7 equal fractions of X rays. Using breathing rate and lethality to measure lung injury up to 1 year after radiation, significant protection against both pneumonitis at 7 months and fibrosis at 12 months was observed using 300 mg/kg of WR-2721. The degree of radioprotection was similar for pneumonitis and fibrosis and was not less after doses per fraction of 4.0 Gy. These data indicate that protection of mouse lung by WR-2721 will not be less in a multifractionated schedule of radiation, at least for doses per fraction greater than 4.0 Gy. PMID- 2982774 TI - Vaccination against feline viral rhinotracheitis in kittens with maternally derived feline viral rhinotracheitis antibodies. AB - The efficacy of a modified live-virus intranasal vaccine and a killed-virus adjuvanted parenteral vaccine in inducing protective immunity against feline viral rhinotracheitis (FVR) was evaluated in kittens with and without maternally derived FVR antibodies. The intranasal vaccine was given as a single dose to kittens 5 weeks old, and the parenteral vaccine was administered in 2 doses at 5 and 7 weeks of age. Seroconversion was delayed for 5 to 10 days in kittens with maternally derived antibodies, but occurred in all vaccinated kittens by 8 weeks of age. When virulent FVR virus was given, both vaccines provided satisfactory protection against disease but did not prevent infection. The results indicated that the modified live-virus intranasal vaccine or the killed-virus adjuvanted parenteral vaccine can be used successfully in kittens with residual maternally derived FVR antibodies. PMID- 2982775 TI - Inclusions in equine cytologic specimens. AB - Inclusions and cellular changes were seen in cytologic specimens from 1 healthy horse, 6 horses hospitalized because of respiratory problems, and 1 horse hospitalized because of colic and hepatitis. Two bronchial aspirates contained detached ciliated cytoplasmic tufts and cytoplasmic inclusions characteristic of the specific degenerative process called ciliocytophthoria. These changes and inclusions resembled those seen in bronchial aspirates from human beings with parainfluenza virus infection. Four bronchial aspirates and 2 serous fluid specimens had nuclear inclusions resembling those seen in a variety of specimens from human beings with herpetic infections. The abnormalities and inclusions were similar to changes seen in cell monolayer and suspension cultures infected with equine herpesvirus type 1. PMID- 2982776 TI - Economic impact of an epizootic of bovine vesicular stomatitis in California. AB - A recent outbreak of vesicular stomatitis in California's San Joaquin Valley caused economic loss at 2 dairies of $225,000 during a 2-month period. These losses amounted to $202/cow for dairy 1 and $97/cow for dairy 2. The most notable economic losses were associated with high cull rates. The rapid spread of the disease (attack rates were 72% in 66 days for dairy 1 and 38% in 41 days for dairy 2) suggests that high-density herds particularly may be vulnerable to the disease. Factors that may have accounted for this rapid spread included common water troughs, open corrals, and inability of the dairy operator to isolate cows due to lack of space. PMID- 2982777 TI - Hypoalbuminemia-related platelet hypersensitivity in two dogs with nephrotic syndrome. AB - Platelet aggregation studies in 2 dogs with nephrotic syndrome disclosed increased platelet sensitivity to a low dose of adenosine diphosphate. Subsequent studies with isolated platelets and plasma indicated that a plasma factor was responsible primarily for inducing platelet hypersensitivity. The increased platelet aggregation response was corrected by increasing the albumin concentration of the plasma. The study suggested an important role for albumin in modulating platelet aggregation and may partially explain the tendency toward thrombosis noted in hypoalbuminemic dogs with nephrotic syndrome. PMID- 2982779 TI - Synthesis of deuterium- and tritium-labeled 6 beta-bromopenicillanic acid. AB - The synthesis of 6 beta-bromopenicillanic acid labeled with deuterium and tritium in the beta-methyl group is described. The S-sulfoxide of benzyl- or p methoxybenzyl 6 alpha-bromopenicillanate is refluxed in benzene containing an excess of tert-BuOD, D2O or HTO. After deoxygenation and deprotection of the ester, the labeled 6 alpha-bromopenicillanic acid is epimerized (N,O bis(trimethylsilyl)acetamide/1,5-diazabicyclo[4.3.0]non-5-ene in CH2Cl2). The two epimers are separated by column chromatography. PMID- 2982778 TI - Testicular LH receptors during aging in Fisher 344 rats. AB - Levels of serum LH, prolactin, testosterone, progesterone and 17-OH progesterone and the testicular concentration and total content of LH receptors were measured in 4-, 11-, 18-, and 27-month-old Fisher 344 rats. All 27-month-old rats had Leydig cell tumors. At first, testicular LH receptor levels decreased with age, but with the appearance of the testicular tumors, these levels increased dramatically. Serum prolactin levels fluctuated with age, but were significantly decreased in 27-month-old rats, as were serum LH levels. Serum testosterone levels decreased steadily with age, while the testosterone-LH receptor ratio remained constant until the appearance of the testicular tumors, after which the ratio decreased precipitously. Serum progesterone levels remained constant throughout the life of Fisher 344 rats until the appearance of testicular tumors, when they increased dramatically. Serum 17-OH progesterone levels were increased significantly at 11 and 27 months as compared to four months of age, but levels at 18 months were similar to those seen in the 4-month-old animals. Therefore, in aged Fisher 344 rats with spontaneous Leydig cell tumors, there is an alteration in the testicular testosterone synthesizing pathway at a step after progesterone. PMID- 2982780 TI - Pituitary concentrations of gonadotropins and receptors for GnRH in suckled beef cows at various intervals after calving. AB - Mature beef cows were slaughtered at 5 (n = 6), 10 (n = 6), 20 (n = 6) or 30 (n = 5) d after calving to identify endocrine events that may affect the duration of postpartum anestrus. Additional cows (n = 6) were slaughtered 12 to 14 d after their first postpartum estrus (luteal phase cows). Anterior pituitary concentrations of luteinizing hormone (LH) were low at d 5 (383 +/- 69 micrograms/g), averaged 445 +/- 103 and 682 +/- 207 micrograms/g at d 10 and 20, respectively, and were elevated (P less than .05) by d 30 (1,097 +/- 174 micrograms) to a concentration similar to luteal phase cows (1,208 +/- 148 micrograms/g). Concentrations of follicle-stimulating hormone (FSH) averaged 12.4 +/- 1.1, 9.6 +/- 2, 8.6 +/- 1.8 and 7.4 +/- 3.3 mg/g at d 5, 10, 20 and 30, respectively. Affinity (1.6 +/- .2 X 10(9) M-1) of anterior pituitary receptors for the GnRH (gonadotropin-releasing hormone) analog (DAla6; des-Gly10, [D-Ala6] LH-RH ethylamide) and weights (2.1 +/- .1 g) of the anterior pituitaries did not differ among groups (P greater than .05). Number of receptors for GnRH averaged 37 +/- 7, 39 +/- 9, 25 +/- 5 and 23 +/- 5 X 10(-14) M/mg protein at d 5, 10, 20 and 30, respectively. Anterior pituitaries from luteal phase cows contained 22 +/ 2 X 10(-14) M/mg protein of receptors for GnRH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982781 TI - Molecular cloning of tetracycline resistance genes from Streptomyces rimosus in Streptomyces griseus and characterization of the cloned genes. AB - Two tetracycline resistance genes of Streptomyces rimosus, an oxytetracycline producer, were cloned in Streptomyces griseus by using pOA15 as a vector plasmid. Expression of the cloned genes, designated as tetA and tetB was inducible in S. griseus as well as in the donor strain. The tetracycline resistance directed by tetA and tetB was characterized by examining the uptake of tetracycline and in vitro polyphenylalanine synthesis by the sensitive host and transformants with the resultant hybrid plasmids. Polyphenylalanine synthesis with crude ribosomes and the S150 fraction from S. griseus carrying the tetA plasmid was resistant to tetracycline, and, by a cross-test of ribosomes and S150 fraction coming from both the sensitive host and the resistant transformant, the resistance directed by tetA was revealed to reside mainly in crude ribosomes and slightly in the S150 fraction. However, the resistance in the crude ribosomes disappeared when they were washed with 1 M ammonium chloride. These results suggest that tetA specified the tetracycline resistance of the machinery for protein synthesis not through ribosomal subunits, but via an unidentified cytoplasmic factor. In contrast, S. griseus carrying the tetB plasmid accumulated less intracellular tetracycline than did the host, and the protein synthesis by reconstituting the ribosomes and S150 fraction was sensitive to the drug. Therefore, it is conceivable that tetB coded a tetracycline resistance determinant responsible for the reduced accumulation of tetracycline. PMID- 2982782 TI - Drug resistance gene amplification of plasmid NR1 derivatives with various amounts of resistance determinant DNA. AB - Drug resistance gene amplification of derivatives of plasmid NR1 having various amounts of resistance (r) determinant DNA was examined with two types of NR1 derivatives. The first was an NR1 derivative that carried two tandem copies of the r determinant component which was isolated as an intermediate in the amplification process. The plating efficiency of host cells and restriction endonuclease analysis of the plasmid DNA indicate that plasmids with two tandem copies of the r determinant undergo spontaneous amplification to a more highly amplified state at a frequency 150-fold higher than that of wild-type NR1. The second class of derivatives consisted of plasmids in which different regions of the r determinant component had been deleted. The relationship between spontaneous amplification frequency and r determinant size was examined with these plasmids. Plating efficiency of host cells indicated that plasmids with a smaller r determinant undergo spontaneous amplification at a lower frequency than do plasmids with a larger r determinant. These results suggest that there is an ordered sequence of events in the amplification of the r determinant of NR1. PMID- 2982783 TI - Characterization of Bacteroides ovatus plasmid pBI136 and structure of its clindamycin resistance region. AB - Genetic and physical analyses were used to characterize the Bacteroides ovatus R plasmid pBI136. Results from restriction endonuclease cleavage studies were used to construct a physical map of the plasmid for the enzymes EcoRI, BamHI, ClaI, XbaI, SalI, and SmaI. Based on the sizes of restriction fragments generated in these studies, the plasmid was estimated to be 80.6 kilobase pairs (kb). A 7.2-kb region of the plasmid required for resistance to lincosamide and macrolide (LM) antibiotics was mapped by analysis of spontaneously occurring LM-sensitive deletion derivatives. Hybridization studies showed that this region and an adjoining 2.9-kb EcoRI fragment were responsible for the previously reported homology among Bacteroides plasmids pBF4, pBFTM10, and pBI136. Within this region of homology, 0.5 kb was attributed to a directly repeated sequence thought to bound the LM resistance determinant on pBF4 and pBFTM10. Two pBI136 EcoRI fragments spanning the putative LM resistance region were cloned in Escherichia coli, and heteroduplex analysis of these recombinant plasmids revealed the presence of a 1.2-kb directly repeated sequence. These results suggested that the pBI136 LM resistance determinant resides on an 8.4-kb segment of DNA containing 6.0 kb of intervening DNA sequences bounded by a 1.2-kb directly repeated sequence. PMID- 2982785 TI - Comparison of physical and genetic properties of palindromic DNA sequences. AB - Some viable palindromic DNA sequences were found to cause an increase in the recovery of genetic recombinants. Although these palindromes contained no Chi sites, their presence in cis caused apparent recA+-dependent recombination to increase severalfold. This biological property did not correlate with the physical properties of the palindromes' extrusion of cruciform structures in vitro. Thus, two unrelated palindromes with similar effects on recombination in both Escherichia coli and Pseudomonas syringae displayed quite different kinetics of cruciform formation. In plasmids of native superhelical density, one palindrome underwent rapid cruciform formation at 55 degrees C, whereas the other did not form detectable cruciforms at any temperature. A shorter palindrome with similarly rapid kinetics of cruciform formation did not affect recombination detectably. The lack of a clear relationship between physical and genetic properties was also demonstrated in the case of longer, inviable palindromes. Here we found that the degree of asymmetry required in vivo to rescue a long palindrome from inviability far exceeded that required to kinetically prohibit cruciform extrusion in vitro. PMID- 2982784 TI - Alteration of phospholipid composition by combined defects in phosphatidylserine and cardiolipin synthases and physiological consequences in Escherichia coli. AB - Escherichia coli K-12 derivatives with a common genetic background carrying, either alone or in combination, the pss-1 allele coding for a temperature sensitive phosphatidylserine synthase (A. Ohta and I. Shibuya, J. Bacteriol. 132:434-443, 1977) and cls- for a defective cardiolipin synthase (G. Pluschke et al., J. Biol. Chem. 253:5048-5055, 1978) were constructed. The phospholipid polar headgroup compositions of these strains were significantly different from each other depending on their genotypes and growth temperature, whereas other membrane characteristics such as the total phospholipid content, fatty acid composition, membrane protein profile, and lipopolysaccharide content were practically the same, suggesting that the phenotypes of these strains were the direct consequences of abnormalities in membrane phospholipid composition. The cls pss-1 double mutation caused an unusual accumulation of phosphatidylglycerol with an extremely low content of cardiolipin. The cls mutation alone was found to give a growth defect, and its introduction into a pss-1 mutant resulted in an enhanced temperature sensitivity of growth. Addition to a broth medium of a proper concentration of sucrose, NaCl, Mg2+, or Ca2+ allowed the growth of a pss-1 mutant at otherwise nonpermissive temperature, but a pss-1 cls double mutant required the combined addition of sucrose or NaCl and MgCl2 for full growth at 42 degrees C. The possible mechanisms for these physiological consequences of the mutations are discussed on a molecular basis. The remedial effects of culture supplements allowed the pss-1 mutants to grow at 42 degrees C resulting in enhanced abnormalities of membrane phospholipid composition. PMID- 2982786 TI - Excision and reintegration of the Escherichia coli K-12 chromosomal element e14. AB - The genetic element e14 is a natural component of the Escherichia coli K-12 chromosome. On induction of the SOS pathways, e14 excises as a 14.4-kilobase circle. We report here on the reintegration of e14 into the chromosome of cured (e14 degrees) E. coli K-12 derivatives. Using a Tn10 insertion mutant of e14, we found that reintegration occurred specifically at the locus originally occupied by e14 and with the same orientation. The reintegration event required neither the RecA nor the RecB functions. The attachment site of the free form was located within a 950-base-pair HindIII-AvaI fragment and shared sufficient homology with the host attachment site to form detectable DNA-DNA hybrids. Even though E. coli C and B/5 did not contain e14, they did possess a HindIII restriction fragment that hybridized to the free e14 attachment fragment. E. coli C could be transformed with e14-1272::Tn10, resulting in integration at this site of homology. The Tn10 mutants were also used in mapping the point of e14 attachment. We found the following sequence: fabD purB atte14 umuC. Furthermore, analysis of a recombinant plasmid that contained both the e14 attachment site and the purB locus showed that these two loci occur within 11 kilobases of each other. PMID- 2982787 TI - Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. AB - A mutation of a cloned gene that has been made by introducing a transposon or some other selectable genetic determinant can be crossed into the gene's original replicon by linearizing the cloned DNA and transforming a recB recC sbcB mutant. A number of applications of this method are described with genes of either chromosomal or plasmid origin. PMID- 2982788 TI - Analysis of mobilization elements in plasmids from Shigella flexneri. AB - The mobilization properties of three plasmids were examined after cotransfer from Shigella flexneri to Escherichia coli. The largest plasmid, pCN1, was shown to be a conjugative R factor that could promote its own transfer and allow cotransfer of a 4.1-kilobase plasmid, pCN3; mobilization of the third plasmid, pCN2 (6.3 kilobases), required the presence of both pCN1 and pCN3. Sequences from pCN2 and pCN3 homologous to the bom (basis of mobilization) sites of ColE1 and pBR322 were localized by analysis of site-specific deletion derivatives generated in vivo during the transfer of composite plasmids and were characterized by DNA sequencing. PMID- 2982789 TI - Rapid method for isolation and screening of cytochrome c oxidase-deficient mutants of Saccharomyces cerevisiae. AB - We describe here a new method for the specific isolation of cytochrome c oxidase deficient mutants of Saccharomyces cerevisiae. One unique feature of the method is the use of tetramethyl-p-phenylenediamine as a cytochrome c oxidase activity stain for yeast colonies. The staining of yeast colonies by tetramethyl-p phenylenediamine is dependent upon a functional cytochrome c oxidase and is unaffected by other lesions in respiration. Since the tetramethyl-p phenylenediamine colony staining reaction is rapid and simple, it greatly facilitates both the identification and characterization of cytochrome c oxidase deficient mutants. Another feature of the method, which is made possible by the tetramethyl-p-phenylenediamine colony stain, is the use of an op1 parent strain for the isolation of nuclear pet or mitochondrial mit mutants in specific protein coding genes. A parent strain that carries this marker selects against rho0 or rho- classes of pleiotropic respiratory-deficient mutants, since these are lethal in op1 strains. We have used this method to isolate 123 independently derived cytochrome c oxidase-deficient pet mutants and 300 independently derived mit mutants. PMID- 2982790 TI - Purification and characterization of the flagellar hook-basal body complex of Salmonella typhimurium. AB - The hook-basal body complex of Salmonella typhimurium, a major component of its flagellar apparatus, was subjected to detailed analysis by electron microscopy and gel electrophoresis. The study was facilitated by the development of an improved protocol for isolation of the complexes in high yield and purity. Nine proteins were identified with the structure. These proteins had apparent molecular weights of 65,000 (65K), 60K, 42K, 38K, 32K, 30K, 27K, 16K, and 14K. Small but reproducible shifts in the apparent molecular weights of specific proteins from conditionally nonflagellate mutants indicated the following gene polypeptide correspondences: flaFV, 42K; flaFVI, 32K; flaFVII, 30K; flaFIX, 38K; flaAII.1, 65K. Several new morphological features of hook-basal body complexes were recognized, including a clawlike structure on the cytoplasm-proximal M ring and additional material at the cytoplasmic face of the M ring. Based on this study and the work of others, we suggest that the morphological features of the hook-basal body complex correspond to the following proteins: hook-filament junction, 60K; hook, 42K; rod, 30K and 32K; L ring and outer cylinder wall, 27K; P ring, 38K; S ring, unknown; M ring 65K. PMID- 2982791 TI - Identification and genetic analysis of an Agrobacterium tumefaciens chromosomal virulence region. AB - A genetic analysis of Agrobacterium tumefaciens chromosomal functions required for virulence was undertaken. Large Tn5-containing cosmid clones were isolated from DNA of avirulent A. tumefaciens mutants having chromosomal Tn5 insertions and exhibiting defective attachment to plant cells. The clones from several different mutants each contained overlapping segments of a 30-kilobase A. tumefaciens chromosomal region, which were physically mapped. All chromosomal Tn5 insertions leading to the avirulent, attachment-defective phenotype were localized within an 11-kilobase portion of this chromosomal virulence region. Transposon Tn3::HoHo1 (Tn3 containing lacZ) was used to simultaneously mutagenize and create lac fusions within the virulence region. This analysis demonstrated the presence of two distinct chromosomal virulence loci, which were 1.5 and 5 kilobases long; transposon insertions into these loci led to avirulence and defective attachment. The beta-galactosidase activity associated with various Tn3::HoHo1-created lac fusions indicated that the loci are transcribed in opposite directions, and complementation studies suggested that each locus consists of a single transcriptional unit. A cosmid clone of the chromosomal virulence region containing a lac fusion in the extreme 3' portion of the 5 kilobase locus was used to demonstrate that expression of this region is dependent on the presence of sequences in the 5' portion of the locus, confirming its operon-like nature. PMID- 2982793 TI - Cloning and expression of the gene for the vitamin B12 receptor protein in the outer membrane of Escherichia coli. AB - The transport of cyanocobalamin (vitamin B12) in cells of Escherichia coli is dependent on a receptor protein (BtuB protein) located in the outer membrane. A 9.1-kilobase pair BamHI fragment carrying the btuB gene was cloned from a specialized transducing phage into multicopy plasmids. Insertions of transposon Tn1000 which prevented production of the receptor localized btuB to a 2-kilobase pair region. Further subcloning allowed isolation of this region as a 2.3 kilobase pair Sau3A fragment. The BtuB+ plasmids were shown by maxicell analysis to encode a polypeptide with a molecular weight of 66,000 in the outer membrane. This polypeptide was missing in cells with Tn1000 insertions in btuB and was reduced in amount upon growth of plasmid-bearing cells in repressing concentrations of vitamin B12. Several Tn1000 insertions outside the 5' end of the coding region exhibited reduced production of receptor. A deletion at the 3' end of btuB resulted in formation of an altered receptor. Amplified production of this polypeptide was associated with increased levels of binding of the receptor's ligands (vitamin B12 and phage BF23), increased rates of vitamin B12 uptake, and altered susceptibility to the group E colicins. Deficiency in various major outer membrane proteins did not affect production of the btuB product, and the amplified levels of this protein partially reversed the tolerance to E colicins seen in these mutants. PMID- 2982792 TI - Induction and autoregulation of ada, a positively acting element regulating the response of Escherichia coli K-12 to methylating agents. AB - The ada gene of Escherichia coli K-12, the regulatory locus for the adaptive response to methylating agents, coded for a 39,000-dalton protein. An adjacent gene coding for a 27,000-dalton protein was coregulated with ada. The Ada protein was strongly induced upon exposure of cells to methylating agents such as N methyl-N'-nitro-N-nitrosoguanidine. An analysis of ada regulation with an ada lacZ operon fusion showed that ada+ function was required for induction of ada transcription. Derivatives of the ada gene truncated from the 3' end produced proteins which were more potent in stimulating transcription than the product of the intact ada gene, indicating that the transcription-activating function of the Ada protein resided in its amino terminus. The sequence of the ada-regulatory region and the identification of the start site of ada transcription are also presented. PMID- 2982794 TI - Molecular cloning in Escherichia coli of Erwinia chrysanthemi genes encoding multiple forms of pectate lyase. AB - The phytopathogenic enterobacterium Erwinia chrysanthemi excretes multiple isozymes of the plant tissue-disintegrating enzyme, pectate lyase (PL). Genes encoding PL were cloned from E. chrysanthemi CUCPB 1237 into Escherichia coli HB101 by inserting Sau3A-generated DNA fragments into the BamHI site of pBR322 and then screening recombinant transformants for the ability to sink into pectate semisolid agar. Restriction mapping of the cloned DNA in eight pectolytic transformants revealed overlapping portions of a 9.8-kilobase region of the E. chrysanthemi genome. Deletion derivatives of these plasmids were used to localize the pectolytic genotype to a 2.5-kilobase region of the cloned DNA. PL gene expression in E. coli was independent of vector promoters, repressed by glucose, and not induced by galacturonan. PL accumulated largely in the periplasmic space of E. coli. An activity stain used in conjunction with ultrathin-layer isoelectric focusing resolved the PL in E. chrysanthemi culture supernatants and shock fluids of E. coli clones into multiple forms. One isozyme with an apparent pI of 7.8 was produced at a far higher level in E. coli and was common to all of the pectolytic clones. Activity staining of renatured PL in sodium dodecyl sulfate-polyacrylamide gels revealed that this isozyme comigrated with the corresponding isozyme produced by E. chrysanthemi. The PL isozyme profiles produced by different clones and deletion derivative subclones suggest that the cloned region contains at least two PL isozyme structural genes. Pectolytic E. coli clones possessed a limited ability to macerate potato tuber tissues. PMID- 2982795 TI - recA-independent recombination between repeated IS50 elements is not caused by an IS50-encoded function. AB - Certain pBR322-related plasmids containing direct repeats of the insertion element IS50 appear to be unstable in recA Escherichia coli because smaller recombinant derivatives accumulate rapidly in plasmid DNA populations. We show here that (i) this instability is plasmid specific, but not IS50 specific; (ii) it is due to a detrimental effect exerted by these plasmids on bacterial growth; and (iii) the growth impairment is alleviated in cells harboring the smaller recombinant plasmids. Although a recent report had concluded that accumulation of recombinants reflected an IS50-specific recombination function, when correction is made for the relative growth rates of cells containing the parental and recombinant plasmids the evidence for such a recombination function disappears. PMID- 2982796 TI - Construction of broad-host-range cosmid cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol. AB - Four new cloning vectors have been constructed from the broad-host-range cloning vector pRK290. These vectors, pLA2901, pLA2905, pLA2910, and pLA2917, confer resistance to kanamycin and tetracycline. The latter two are cosmid derivatives of pLA2901. The new vectors can be mobilized into, and are stably maintained in, a variety of gram-negative bacteria. A Sau3A genomic bank of Methylobacterium organophilum strain xx DNA has been constructed in pLA2917, and complementation analysis, with a variety of mutants unable to grow on methanol, revealed at least five separate regions necessary for growth on methanol. Complementation analysis and Tn5 mutagenesis data suggest that at least three genes are responsible for expression of active methanol dehydrogenase. PMID- 2982797 TI - Role of proton motive force in phototactic and aerotactic responses of Rhodopseudomonas sphaeroides. AB - Rhodopseudomonas sphaeroides grown under nonrigorous anaerobic conditions in the light developed components of a branched respiratory electron transfer chain, and a photosynthetic electron transfer chain. Both respiratory pathways were sensitive to rotenone and high concentrations of cyanide, but oxygen uptake was only partially inhibited by the addition of low concentrations of cyanide or antimycin A. When incubated anaerobically in the dark, R. sphaeroides responded positively to an oxygen gradient in the absence of rotenone. In the presence of rotenone, aerotaxis only occurred when the antimycin A-sensitive branch of the pathway was functioning, although both branches still reduced oxygen. Although there was electron movement along the respiratory chain, aerotaxis only occurred in response to a change in proton motive force. When incubated anaerobically in the light, the movement of R. sphaeroides up a light gradient depended on photosynthetic electron transport. When incubated aerobically, high-intensity actinic illumination inhibited oxygen uptake and aerotaxis. In a low-intensity light gradient the phototactic response was inhibited by oxygen. These results are discussed in relation to the interaction of the electron transfer chains and their roles in controlling tactic responses in R. sphaeroides. PMID- 2982798 TI - Sequence of human asialoglycoprotein receptor cDNA. An internal signal sequence for membrane insertion. AB - A cDNA library from the human hepatoma cell line Hep G2 was prepared in the expression vector lambda gt11. Using specific antibodies, a cDNA clone containing the entire coding sequence for the human asialoglycoprotein receptor was isolated and sequenced. The deduced amino acid sequence of 291 residues is very homologous to the sequence of the major asialoglycoprotein receptor protein from rat. The comparison shows that there is no significant post-translational processing and no leader sequence, cleaved or uncleaved, at the amino terminus. An internal signal sequence, probably the membrane-spanning segment, residues 41-59, is assumed to direct insertion of the carboxyl-terminal ligand binding portion of the receptor across the endoplasmic reticulum membrane. PMID- 2982799 TI - Purification and crystallization of the EcoRV restriction endonuclease. AB - The type II restriction endonuclease EcoRV purified from a genetically engineered, overproducing strain has been crystallized. Four crystal forms all obtained by precipitation with polyethylene glycol 4000 have been characterized. Two of these are suitable for high resolution structure analysis. Both are orthorhombic, have space group P2(1)2(1)2(1) and have similar unit cell dimensions of a = 58.2 A, b = 71.7 A, c = 130.6 A (form A) and a = 59.9 A, b = 74.5 A, c = 121.8 A (form B). They diffract to about 2A resolution and appear to have one dimer of 2 X 29,000 daltons in the asymmetric unit. PMID- 2982800 TI - Phosphorylation of the erbB gene product from an avian erythroblastosis virus transformed chick fibroblast cell line. AB - Polyclonal antiserum prepared against the human epidermal growth factor receptor immunoprecipitated four proteins of Mr = 66,000, 68,000, 74,000, and 82,000 from avian erythroblastosis virus-transformed chick embryo fibroblasts (cell line AEV C23) which seemed to be related to the erbB gene product. The Mr = 66,000 and 68,000 proteins chased into the Mr = 74,000 and 82,000 proteins in pulse-chase experiments. The Mr = 68,000 and 82,000 proteins were found to be phosphorylated primarily on serine and threonine residues and contained minor amounts of phosphotyrosine. Tryptic peptide analysis of these phosphoproteins revealed several major peptides, and treatment of cells with the tumor promoter 12-O tetradecanoyl-phorbol-13-acetate resulted in the appearance of an additional phosphopeptide. 12-O-Tetradecanoyl-phorbol-13-acetate also inhibited growth of AEV-C23 cells in soft agar and in monolayer culture. In vitro phosphorylation of Mr = 68,000 and 74,000 proteins in immunoprecipitates occurred on tyrosine with lesser amounts of phosphoserine and phosphothreonine detected. PMID- 2982801 TI - Insulin and epidermal growth factor do not affect phosphoinositide metabolism in rat liver plasma membranes and hepatocytes. AB - Recent studies with viral oncogene tyrosine kinases have suggested that these kinases may phosphorylate phosphoinositides and diacylglycerol. Since the receptors for insulin and epidermal growth factor (EGF) also possess tyrosine kinase activity, we have investigated possible effects of insulin and EGF on phosphoinositide metabolism in rat liver plasma membranes and rat hepatocytes. In plasma membranes prepared from rats injected 18 h prior with [3H]myo-inositol or incubated with [gamma-32P]ATP, phosphatidylinositol-4-P and phosphatidylinositol 4,5-P2 were formed, but there were no effects of either insulin or EGF although these agents stimulated protein tyrosine phosphorylation. In hepatocytes incubated with [3H]myo-inositol, label was incorporated into phosphatidylinositol, phosphatidylinositol-4-P, and phosphatidylinositol-4,5-P2, but there was no effect of insulin. Incubation of hepatocytes with [3H]myo inositol plus insulin or EGF for 2 h also did not alter the formation of [3H]myo inositol-1,4,5-P3 from [3H]phosphatidylinositol-4,5-P2 induced by vasopressin. These findings suggest that the tyrosine kinase activity of liver insulin and EGF receptors is not important in phosphoinositide formation. PMID- 2982802 TI - ADP stimulates hydrolysis of the "ADP-insensitive" phosphoenzyme in Na+, K+ ATPase and Ca2+-ATPase. AB - ADP-sensitive (E1P) and K+-sensitive (E2P) phosphoenzymes are sequentially formed intermediates in the reaction pathways catalyzed by the Na+,K+- and Ca2+-ATPases. The kinetics of dephosphorylation of these intermediates were examined by means of rapid quenching with acid at 21 degrees C. Under conditions favoring the formation of E2P (25 mM Na+ and O K+), addition of 5 mM ADP + 10 mM EDTA to the Na+,K+-ATPase phosphoenzyme produced a biphasic pattern of dephosphorylation. Both phases of phosphoenzyme decomposition were accompanied by approximately stoichiometric amounts of inorganic phosphate (Pi) release. The rate of decay of the rapid phase was 10 times faster than the rate of phosphoenzyme turnover under phosphorylating conditions indicating acceleration of E2P hydrolysis by ADP. Similar patterns of ADP-stimulated phosphoenzyme decay and Pi release were observed in the Ca2+-ATPase from sarcoplasmic reticulum phosphorylated at low (0.1 mM) Mg2+ in the absence of KCl. These results demonstrate that ADP can enhance the rate of E2P hydrolysis in the cases of the Na+,K+-ATPase and Ca2+ ATPase. As a consequence measurement of "ADP-sensitive EP" may overestimate E1P. PMID- 2982803 TI - Specificity of protein phosphotyrosine phosphatases. Comparison with mammalian alkaline phosphatase using polypeptide substrates. AB - The specificity of cytosolic protein phosphotyrosine (PPT) phosphatases was investigated using different peptides and proteins that were phosphorylated on tyrosine residues by the EGF receptor kinase. The acidic phosphoproteins, serum albumin, casein, and myosin light chains, were dephosphorylated by the PPT phosphatases with apparent Km values of 1.2 to 12.5 microM and apparent velocities of 0.2 to 18 mumol/min/mg. In contrast, [Tyr(32P)]histone and the phosphotyrosine peptides [Val5]angiotensin and RR-src, a peptide with sequence Arg-Arg-Leu-Ile-Glu-Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Gly, were unreactive with the PPT phosphatases. However, each of these unreactive phosphopolypeptides was dephosphorylated under the same conditions by calf-intestine alkaline phosphatase. The data reveal how PPT phosphatase activity has been ascribed to different cellular enzymes. When acidic phosphotyrosine proteins were used as substrates in assays for PPT phosphatase activity the cytosolic enzymes were isolated, whereas when phosphotyrosine histones were used as substrates only the membrane-bound alkaline phosphatase was detected. Apparently the protein tyrosine kinase and the protein tyrosine phosphatases do not have the same specificity, so substrates such as histone, angiotensin, or RR-src are phosphorylated but not hydrolyzed. Therefore, these polypeptides would be ideal for the characterization of protein tyrosine kinases in cellular extracts. PMID- 2982804 TI - Subunit phosphorylation and activation of skeletal muscle phosphorylase kinase by the cAMP-dependent protein kinase. Divalent metal ion, ATP, and protein concentration dependence. AB - This report provides a characterization of the effects of varying the concentrations of Mg2+, ATP, phosphorylase kinase, and the cAMP-dependent protein kinase on the activation and phosphorylation of phosphorylase kinase. The results show the following. (a) The Km for MgATP2- for the cAMP-dependent protein kinase catalyzed phosphorylation is decreased by increasing Mg2+, probably as a consequence of decreasing the free ATP:MgATP2- ratio and increasing free Mg2+. (b) Whereas beta subunit phosphorylation of phosphorylase kinase plays a prominent role in determining its activity, alpha subunit phosphorylation can also modulate activity. (c) The phosphorylation of the alpha subunit, which occurs following the initial cAMP-dependent phosphorylation of the beta subunit, is catalyzed by the cAMP-dependent protein kinase and is not a consequence of EGTA-insensitive (or EGTA-sensitive) autophosphorylation occurring as a result of the enhanced phosphorylase kinase activity. (d) The relationship between subunit phosphorylation and phosphorylase kinase activation is complex and particularly dependent upon concentrations of cAMP-dependent protein kinase and phosphorylase kinase in the activation reaction. The data suggest the possibilities that the pathway of phospho-intermediates involved in the activation process probably varies with the activation conditions, that the efficacy of a specific site to be covalently modified is dependent upon the phosphorylation status of other sites, and that the effect of phosphorylation in regulating activity may also be dependent on the phosphorylation status of other sites. It is clear from the data that the activation process for phosphorylase kinase can be very complex, and it is possible that this complexity might have significant physiological ramifications. PMID- 2982805 TI - The inhibitory guanine nucleotide-binding protein (Ni) purified from bovine brain is a high affinity GTPase. AB - Using modifications of the methods of Bokoch et al. (Bokoch, G.M., Katada, T., Northup, J. K., Ui, M., and Gilman, A. G. (1984) J. Biol. Chem. 259, 3560-3567) and Codina et al. (Codina, J., Hildebrandt, J. D., Sekura, R. D., Birnbaumer, M., Bryan, J., Manclark, C. R., Iyengar, R., and Birnbaumer, L. (1984) J. Biol. Chem. 259, 5871-5886), we have purified a pertussis toxin substrate with the expected characteristics of the inhibitory guanine nucleotide-binding protein (Ni) essentially to homogeneity. The purified protein consists of 3 subunits of Mr 40,000, 35,000, and less than 10,000. The Mr 40,000 band is found, upon close examination, to consist of a poorly resolved doublet. Starting with the membranes from 1,320 g of bovine forebrain we purified the protein some 100-fold with approximately 20% yield to obtain 13 mg of a greater than 95% pure protein. Chromatography on octyl-Sepharose provided efficient separation of Ni from Ns (the stimulatory guanine nucleotide-binding protein). Analytical ultracentrifugation indicates an Mr of 82,000 and a sedimentation coefficient S20,w of 5.1. The protein is able to restore opiate-mediated inhibition of adenylate cyclase to membranes prepared from NG 108-15 cells which had been treated with pertussis toxin. Bovine brain Ni has the enzymatic properties of a low Km GTPase with a turnover number of 0.3 and affinities for nucleotides in the order GppNHp greater than or equal to GTP greater than or equal to GDP much greater than ATP, CTP, UTP, and GMP. Na+ specifically stimulates the GTPase and low concentrations of Mg2+ (less than 50 microM) are inhibitory. Some Mg2+ is apparently necessary because EDTA, but not EGTA, abolishes the GTPase activity. PMID- 2982806 TI - Differential selectivity of cholinephosphotransferase and ethanolaminephosphotransferase of Tetrahymena for diacylglycerol and alkylacylglycerol. AB - The glycerophospholipids of the ciliate protozoan Tetrahymena thermophila differ greatly in their content of alkylacylglycerol with phosphatidylcholine, phosphatidylethanolamine, and 2-aminoethylphosphonolipid containing 60, 4, and 53% glyceryl ether, respectively. This difference is achieved by differences in the selectivities of cholinephosphotransferase (EC 2.7.8.2) and ethanolaminephosphotransferase (EC 2.7.8.1) for alkylacylglycerol and diacylglycerol. When the two enzymes are assayed in vitro using only endogenous diglyceride as substrate, the newly formed phosphatidylcholine contains 37% glyceryl ether, while the newly formed phosphatidylethanolamine contains 5% glyceryl ether. The ethanolaminephosphotransferase is stimulated equally well by addition of diolein and dipalmitin, but the diacylglycerols have no effect on the glyceryl ether content of phosphatidylethanolamine. In contrast, the glyceryl ether content of newly formed phosphatidylcholine decreases to 16% when the cholinephosphotransferase is exposed to diolein or dipalmitin. The ethanolaminephosphotransferase is not stimulated by addition of a 60:40 mixture of alkylacylglycerol/diacylglycerol. The cholinephosphotransferase is stimulated by the mixture to the same extent as it is by the diacylglycerols, with the glyceryl ether content of the newly formed phosphatidylcholine increasing to 52%. With the addition of alkylacylglycerol alone, the glyceryl ether content of the newly formed phosphatidylethanolamine increases to 10%, while that of the newly formed phosphatidylcholine increases almost to 60%. PMID- 2982807 TI - Stimulation of collagenase synthesis by a 20,000-dalton epithelial cytokine. Evidence for pretranslational regulation. AB - Certain products of cultured rabbit corneal epithelium or epidermis regulate collagenase production by rabbit corneal stromal cells. Here, we examined the effects of a 20-kDa cytokine derived from cultured epithelium on collagenase expression by normal human skin fibroblasts and by cells from recessive dystrophic epidermolysis bullosa, a disease characterized by over-production of a structurally altered collagenase. Culturing cells for 24 h in the presence of the cytokine resulted in an approximately 2-fold increase in trypsin-activatable collagenase activity paralleled by an increase in immunoreactive protein, suggesting enhanced synthesis. There was no change in the activity/immunoreactive protein, indicating a catalytically unaltered enzyme. In kinetic studies, stimulation of collagenase synthesis was first observed approximately 8 h after exposure to the epidermal cytokine. Conversely, when cells were primed with cytokine for 24 h and the stimulator was then removed, an increased rate of synthesis was seen for an additional approximately 9 h, after which the rate reverted to control levels. Since the kinetic data suggested an effect at a pretranslational level, fibroblasts cultured with 20-kDa stimulator were used to prepare mRNA. In cell-free translation total protein synthesis was unaltered; however, the cytokine caused a greater than 2-fold increase in translatable collagenase mRNA. The data suggest that this epithelial cytokine specifically modulates collagenase synthesis through transcription and that epithelial-stromal interactions are important to cutaneous connective tissue metabolism. PMID- 2982808 TI - Steroid product-induced, oxygen-mediated damage of microsomal cytochrome P-450 enzymes in Leydig cell cultures. Relationship to desensitization. AB - Treatment of Leydig cells with 1 mM 8-Br-cAMP for 48 h decreased microsomal cytochrome P-450 activities, 17 alpha-hydroxylase and C17-20 lyase, by 60-75% and resulted in desensitization of the steroidogenic response. Reduction of the oxygen tension from 19 to 1% O2 prevented the decrease in P-450 activities but not the reduction in steroidogenic capacity. The decrease in activity was also prevented by blocking steroid synthesis with aminoglutethimide. Treatment of cultures with steroid products, androstenedione or testosterone, or product analogs, epitestosterone or 17 alpha-methyltestosterone, at a concentration of 2 microM, which is equivalent to the concentration of testosterone resulting from stimulation with cAMP, also caused oxygen tension-sensitive decreases in hydroxylase activity. Treatment of cultures with 2 microM cortisol, estradiol, or methyltrienolone, an androgen receptor agonist, did not decrease hydroxylase activity, nor did treatment with an androgen receptor antagonist prevent the cAMP or testosterone-induced decreases in hydroxylase activity. Reductions in hydroxylase and lyase activities resulting from testosterone- or epitestosterone treatment had little or no effect on acute cAMP-stimulated testosterone production, whereas desensitization with cAMP caused an 80-90% reduction in steroidogenic capacity. 22R-Hydroxycholesterol-supported testosterone synthesis was decreased by both cAMP- and steroid-treatment at 19% O2, but not below the cAMP-stimulated level. Reduction of the oxygen tension partially prevented this decrease. These data are consistent with the hypothesis that the decline in microsomal P-450 enzymes in desensitized Leydig cells results from product (pseudosubstrate)-induced, oxygen-derived, free-radical damage rather than a steroid receptor-mediated process. PMID- 2982809 TI - Modulation of muscle phosphofructokinase at physiological concentration of enzyme. AB - Two approaches have been used to study the allosteric modulation of phosphofructokinase at physiological concentration of enzyme; a "slow motion" approach based on the use of a very low Mg2+/ATP ratio to conveniently lower Vmax, and the addition of polyethylene glycol as a "crowding" agent to favor aggregation of diluted enzyme. At 0.6 mg/ml muscle phosphofructokinase exhibited a drastic decrease in the ATP inhibition and the concomitant increase in the apparent affinity for fructose-6-P, as compared to a 100-fold diluted enzyme. Similar results were obtained with diluted enzyme in the presence of 10% polyethylene glycol (Mr = 6000). Results with these two approaches in vitro were essentially similar to those previously observed in situ (Aragon, J. J., Feliu, F. E., Frenkel, R., and Sols, A. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 6324 6328), indicating that the enzyme is strongly dependent on homologous interactions at physiological concentrations. With polyethylene glycol it was observed that within the physiological range of concentration of substrates and the other positive effectors, fructose-2,6-P2 still activates the liver phosphofructokinase although it no longer significantly affects the muscle isozyme. In the presence of polyethylene glycol, muscle phosphofructokinase can approach its maximal rate even in the presence of physiologically high concentrations of ATP. Three minor activities of muscle phosphofructokinase have been studied at high enzyme concentration: the hydrolysis of MgATP (ATPase) and fructose-1,6-P2 (FBPase), produced in the absence of the other substrate, and the reverse reaction from MgADP and fructose-1,6-P2. The kinetic study of these activities has allowed a new insight into the mechanisms involved in the modulation of phosphofructokinase activity. The binding of (Mg)ATP at its regulatory site reduces the ability of the enzyme to cleave the bond of the terminal phosphate of MgATP at the substrate site. The positive effectors (Pi, cAMP, NH+4, fructose-1,6-P2, and fructose-2,6-P2) decrease the inhibitory effect of MgATP. Citrate and fructose-2,6-P2 both act as mechanistically "secondary" effectors in the sense that citrate does not inhibit and fructose-2,6-P2 does not activate the FBPase activity, requiring both the presence of ATP to affect the enzyme activity. In conclusion it appears that the regulatory behavior of mammalian phosphofructokinases is utterly dependent on the fact of their high concentrations in vivo. PMID- 2982810 TI - Phorbol esters inhibit the fMet-Leu-Phe- and leukotriene B4-stimulated calcium mobilization and enzyme secretion in rabbit neutrophils. AB - The tumor co-promoter phorbol 12, myristate 13, acetate (PMA) has previously been shown to stimulate several of the characteristic functions (aggregation, degranulation, and the oxidative burst) of polymorphonuclear leukocytes (neutrophils). We describe here a novel feature of the action of PMA on neutrophils, namely its ability to inhibit the chemotactic factor-induced increased in the enzyme secretion and in the intracellular concentration of free calcium. The inhibition is maximal within 3 min of the addition of PMA and is concentration-dependent (IC50 = 8.5 ng/ml). The site of action of PMA is distal to the binding of the chemotactic factors. PMA inhibits both the release of intracellular calcium and the permeability changes to calcium induced by chemotactic factors, but does not affect the stimulation of the rate of influx of sodium produced by the same agents. The PMA analog 4 alpha-phorbol 12, 13 didecanoate, which lack tumorigenicity and the ability to activate the calcium- and phospholipid-dependent protein kinase (protein kinase C), does not inhibit any of the above fMet-Leu-Phe-stimulated neutrophil functions. The present results thus demonstrate that phorbol esters, either directly or indirectly, possibly through the activation of protein kinase C, inhibit the signal(s) responsible for the stimulated mobilization of calcium in rabbit neutrophils. PMID- 2982811 TI - Desensitization of the turkey erythrocyte beta-adrenergic receptor in a cell-free system. Evidence that multiple protein kinases can phosphorylate and desensitize the receptor. AB - We have used a recently developed cell-free system (cell lysate) derived from turkey erythrocytes to explore the potential role of cAMP-activated and other protein kinase systems in desensitizing the adenylate cyclase-coupled beta adrenergic receptor. Desensitization by the agonist isoproterenol required more than simple occupancy of the receptor by the agonist since under conditions where adenylate cyclase was not activated, no desensitization occurred. As in whole cells, addition of cyclic nucleotides to the cell lysate produced only approximately 50% of the maximal isoproterenol-induced desensitization obtainable. Addition of the purified cAMP-dependent protein kinase holoenzyme plus isoproterenol to isolated turkey erythrocyte plasma membranes mimicked the submaximal desensitization induced in lysates by cAMP. This effect was entirely blocked by the specific inhibitor of the cAMP-dependent protein kinase. By contrast, maximal desensitization induced in lysates by isoproterenol was only approximately 50% attenuated by the protein kinase inhibitor. In the lysate preparations, isoproterenol was also shown to induce, in a stereospecific fashion, phosphorylation of the beta-adrenergic receptor. Phosphorylation promoted by isoproterenol was attenuated by cAMP-dependent protein kinase inhibitor to the same extent as desensitization (i.e. approximately 50%). Phorbol diesters also promoted receptor desensitization and phosphorylation in cell lysates. The desensitization was mimicked by incubation of isolated turkey erythrocyte membranes with partially purified preparations of protein kinase C plus phorbol diesters. In the cell lysate, calmodulin also promoted receptor phosphorylation and desensitization which was blocked by EGTA. Desensitization of adenylate cyclase by isoproterenol, phorbol diesters, and calmodulin was not observed to be additive. These findings suggest that: (a) multiple protein kinase systems, including cAMP-dependent, protein kinase C-dependent, and Ca2+/calmodulin-dependent kinases, are capable of regulating beta-adrenergic receptor function via phosphorylation reactions and that (b) cAMP may not be the sole mediator of isoproterenol-induced phosphorylation and desensitization in these cells. PMID- 2982812 TI - Molecular size of the epidermal growth factor receptor-kinase as determined by radiation inactivation. AB - Radiation inactivation with high energy electrons from a linear accelerator was used to determine the functional molecular size of the epidermal growth factor (EGF) binding site and the tyrosine-specific protein kinase activity in A-431 membranes. The target size of the protein portion of the EGF receptor glycoprotein was 147,000 daltons when the radiation-dependent decrease in maximal binding capacity was measured. Since the target size is in good agreement with the molecular size of the protein portion of the EGF receptor determined by denaturing biochemical methods, it appears that the monomeric receptor is the functional binding site in situ. The target size of the EGF-stimulated kinase activity associated with the affinity-purified EGF receptor/kinase was 133,000 and 144,000 daltons when assayed for the ability to autophosphorylate or to phosphorylate a tyrosine-containing peptide, respectively. However, the target size of the kinase activity that did not adhere to an EGF-affinity column was 54,000 and 69,000 daltons when assayed for phosphorylation of endogenous and exogenous substrates, respectively. Intermediate target sizes were obtained when kinase assays were performed on membranes prior to fractionation by affinity chromatography. These results, taken with other biochemical data, indicate that A 431 membranes contain a kinase activity that is a domain of the glycoprotein that contains the EGF binding site and that the membranes also contain another tyrosine-specific kinase or kinases that have an average size of approximately 60,000 daltons. PMID- 2982813 TI - Phosphorylation of type II regulatory subunit of cAMP-dependent protein kinase in intact smooth muscle. AB - A monoclonal antibody was used to quantitate changes in the extent of phosphorylation of the type II regulatory subunit of cAMP-dependent protein kinase in intact bovine tracheal smooth muscle. The autophosphorylated and nonphosphorylated forms of the regulatory subunit (RII) were separated in sodium dodecyl sulfate-polyacrylamide gels and identified by immunoblot analysis. Addition of cAMP to tissue extracts resulted in rapid dephosphorylation of RII (t 1/2 = 20s at 4 degrees C) while addition of MgATP caused complete conversion to the phosphorylated form. Under basal conditions, 56% of RII in intact muscle was phosphorylated when the tissue was homogenized under conditions which fully inhibit protein kinase and phosphatase activities. Incubation with isoproterenol caused a dose-dependent decrease in the phosphorylation state of RII (EC50 = 5 X 10(-8) M). Incubation with high concentrations of isoproterenol, 1-methyl-3 isobutylxanthine, or forskolin caused maximal decreases in the phosphorylated form to 12-18% of the total RII. The effect of isoproterenol was rapid (t 1/2 = 15 s at 37 degrees C), reversible, and could be blocked with the antagonist propranolol. Contraction of the smooth muscle with K+ or low (less than 1 microM) concentrations of carbachol had no effect on the phosphorylation level. A decrease in the basal phosphorylation level to 41% was observed with 10 microM carbachol which was additive with the dephosphorylation produced by isoproterenol. The time course of isoproterenol-induced dephosphorylation of RII paralleled that of muscle relaxation, consistent with a role of cAMP-dependent protein kinase activation in relaxation of smooth muscle. PMID- 2982814 TI - Heterogeneity at the 5' termini of mouse dihydrofolate reductase mRNAs. Evidence for multiple promoter regions. AB - We have determined the sequence of the 1000 base pairs of DNA preceding the coding region of the mouse dihydrofolate reductase gene. 700 base pairs upstream of the translation start codon (position + 1) is the sequence CAACT, separated by 50 base pairs from the sequence TAATAA; these sequences resemble controlling elements responsible for accurate and efficient transcription initiation in a variety of eukaryotic genes. The region between -244 and -101 consists of a 3 fold tandem repeat of a 48-base pair sequence. S1 mapping results indicate that the 5' termini of the multiple dihydrofolate reductase mRNAs are heterogeneous, with the major terminus at -115 and with minor termini in the regions of -275 and -450. In addition, a portion of the 5'-ward region of the gene from -543 to -405 is represented differentially in some, but not all, mouse dihydrofolate reductase mRNAs. The above findings are consistent with the presence of two transcription promoter elements in the 5' end of the mouse dihydrofolate reductase gene. This has been substantiated by inserting portions of the 5'-ward 1000 base pairs of the gene into modular dihydrofolate reductase plasmids. When such constructs are transfected into Chinese hamster ovary cells lacking dihydrofolate reductase, function can be restored with equal transfection frequency when sequences surrounding the 5' CAACT-TAATAA region at -700 or a complete 48-base pair repeat from the region of -148 to -101 are present in the plasmid construct. PMID- 2982815 TI - Purification and properties of ferredoxinTOL. A component of toluene dioxygenase from Pseudomonas putida F1. AB - Toluene dioxygenase oxidizes toluene to (+)-cis-1(S),2(R)-dihydroxy-3 methylcyclohexa-3,5-diene. This reaction is catalyzed by a multienzyme system that is induced in cells of Pseudomonas putida F1 during growth on toluene. One of the components of toluene dioxygenase has been purified to homogeneity and shown to be an iron-sulfur protein that has been designated ferredoxinTOL. The molecular weight of ferredoxinTOL was calculated to be 15,300, and the purified protein was shown to contain 2 g of atoms each of iron- and acid-labile sulfur which appear to be organized as a single [2Fe-2S]cluster. Solutions of ferredoxinTOL were brown in color and showed absorption maxima at 277, 327, and 460 nm. A shoulder in the spectrum of the oxidized protein was discernible at 575 nm. Reduction with sodium dithionite or NADH and ferredoxinTOL reductase resulted in a decrease in visible absorbance at 460 and 575 nm, with a concomitant shift in absorption maxima to 382 and 438 nm. The redox potential of ferredoxinTOL was estimated to be -109 mV. In the oxidized state, the protein is diamagnetic. However, upon reduction it exhibited prominent electron paramagnetic resonance signals with anisotropy in g values (gx = 1.81, gy = 1.86, and gz = 2.01). Anaerobic reductive titrations revealed that ferredoxinTOL is a one-electron carrier that accepts electrons from NADH in a reaction that is mediated by a flavoprotein (ferredoxinTOL reductase). The latter is the first component in the toluene dioxygenase system. Reduced ferredoxinTOL can transfer electrons to cytochrome c or to a terminal iron-sulfur dioxygenase (ISP-TOL) which catalyzes the incorporation of molecular oxygen into toluene and related aromatic substrates. PMID- 2982816 TI - Reversible and irreversible inhibition of phosphate transport in human erythrocytes by a membrane impermeant carbodiimide. AB - Phosphate entry into chloride-loaded human erythrocytes is inhibited by treatment of cells with the water-soluble carbodiimide 1-ethyl-3-(4-azonia-4,4 dimethylpentyl)carbodiimide (EAC) in the absence of added nucleophile. EAC does not penetrate the erythrocyte membrane or lead to significant intermolecular cross-linking of membrane proteins. At neutral extracellular pH in chloride-free medium, only about 50% of transport is rapidly and irreversibly inhibited, but at alkaline pH, inhibition is more rapid and complete. Inhibition by EAC was reversible in the presence of extracellular NaCl. Modification of membrane sulfhydryl groups does not prevent inhibition of phosphate transport by EAC but almost complete protection is afforded by 4,4-dinitrostilbene-2,2-disulfonic acid, a reversible competitive inhibitor of anion transport. N-(4-Azido-2 nitrophenyl)-2-aminoethylsulfonate, a reversible noncompetitive inhibitor of anion transport did not protect against EAC inhibition of transport but prevented reversal of inhibition in saline medium. Transport inhibition by [3H]EAC did not lead to specific incorporation of radioactivity into Band 3, the anion transport protein. These results suggest that inhibition of anion transport by EAC is due to modification of a carboxylic acid residue in or near the transport site accessible from the external face of the membrane. The subsequent fate of the modified carboxyl residue appears to be sensitive to the orientation of the anion transport site. PMID- 2982817 TI - Cytochrome b translocation to human neutrophil plasma membranes and superoxide release. Differential effects of N-formylmethionylleucylphenylalanine, phorbol myristate acetate, and A23187. AB - The role of specific granules and cytochrome b in superoxide (O(2)) release was studied by comparing the effects of three different stimuli on normal human neutrophils, neutrophils congenitally deficient in specific granules, and granule free normal neutrophil cytoplasts. Phorbol myristate acetate (PMA) stimulated normal neutrophils to release more O(2) than did N formylmethionylleucylphenylalanine (fMet-Leu-Phe), which stimulated greater release than the calcium ionophore A23187. Neutrophils lacking specific granules produced variable amounts of O(2) in response to all stimuli. Stimulation with PMA, fMet-Leu-Phe, and A23187 produced maximal rates of O(2) release that were 32, 55, and 21% of that by normal cells. Likewise, granule-free neutrophil cytoplasts released 24, 20, and 0% of the O(2) released by intact cells. These data suggest that the stimuli require different mechanisms for activation. Three subcellular fractions (azurophil granule rich, specific granule rich, and plasma membrane rich) were separated by Percoll gradients from normal resting and stimulated neutrophils. In resting neutrophils, the cytochrome b content in the plasma membrane was 31% of the total, with the rest in the specific granule-rich fraction. Ten minutes after stimulation, PMA, fMet-Leu-Phe, and A23187 induced translocation of 27, 8, and 49%, respectively, of the cytochrome b from the specific granule-rich fraction to the plasma membrane. Although our data support a role for specific granule factors in A23187-induced O(2) release, there is no correlation between the amount of cytochrome b incorporated into the plasma membrane and the extent of O(2) production activated by the different stimuli. PMID- 2982819 TI - Isolation of ubiquinol oxidase from Paracoccus denitrificans and resolution into cytochrome bc1 and cytochrome c-aa3 complexes. AB - An enzyme complex with ubiquinol-cytochrome c oxidoreductase, cytochrome c oxidase, and ubiquinol oxidase activities was purified from a detergent extract of the plasma membrane of aerobically grown Paracoccus denitrificans. This ubiquinol oxidase consists of seven polypeptides and contains two b cytochromes, cytochrome c1, cytochrome aa3, and a previously unreported c-type cytochrome. This c-type cytochrome has an apparent Mr of 22,000 and an alpha absorption maximum at 552 nm. Retention of this c cytochrome through purification presumably accounts for the independence of ubiquinol oxidase activity on added cytochrome c. Ubiquinol oxidase can be separated into a 3-subunit bc1 complex, a 3-subunit c aa3 complex, and a 57-kDa polypeptide. This, together with detection of covalently bound heme and published molecular weights of cytochrome c1 and the subunits of cytochrome c oxidase, allows tentative identification of most of the subunits of ubiquinol oxidase with the prosthetic groups present. Ubiquinol oxidase contains cytochromes corresponding to those of the mitochondrial bc1 complex, cytochrome c oxidase complex, and a bound cytochrome c. Ubiquinol cytochrome c oxidoreductase activity of the complex is inhibited by inhibitors of the mitochondrial bc1 complex. Thus it seems likely that the pathway of electron transfer through the bc1 complex of ubiquinol oxidase is similar to that through the mitochondrial bc1 complex. The number of polypeptides present is less than half the number in the corresponding mitochondrial complexes. This structural simplicity may make ubiquinol oxidase from P. denitrificans a useful system with which to study the mechanisms of electron transfer and energy transduction in the bc1 and cytochrome c oxidase sections of the respiratory chain. PMID- 2982818 TI - Quantitation of apolipoprotein E mRNA in the liver and peripheral tissues of nonhuman primates. AB - A sensitive DNA-excess solution hybridization assay was used to quantitate apo-E mRNA in the liver and peripheral tissues of two nonhuman primates, Macaca fascicularis and Cercopithecus aethiops. When expressed on the basis of total RNA, apo-E mRNA values for M. fascicularis adrenal, brain, testis, and spleen ranged from 17-52% of the liver value. Apo-E mRNA values for mesenteric lymph node, kidney, thymus, and skeletal muscle were 1-5% of the liver value. When expressed on a cellular basis, apo-E mRNA was most abundant in the liver at approximately 1200 molecules/cell. Peripheral tissues showed a continuous range of apo-E mRNA from 1.5 molecules/cell in the thymus up to 350 molecules/cell in the brain. Similar results were obtained with peripheral tissues from C. aethiops in which case apo-E mRNA also was found in skin, lung, skeletal muscle, small intestine, and vascular tissues such as heart, aorta, and brachial artery. Calculation of the total apo-E mRNA/organ showed that most of the apo-E mRNA was present in the liver. However, summation of apo-E mRNA in peripheral tissues indicated that 20-40% of total body apo-E mRNA was extrahepatic. This results indicates that apo-E made in peripheral tissues may play a quantitatively important role in cholesterol metabolism since peripheral tissues have the potential to contribute a significant fraction of plasma apo-E. PMID- 2982820 TI - Only the promoter region of the constitutively expressed normal and amplified human dihydrofolate reductase gene is DNase I hypersensitive and undermethylated. AB - The human dihydrofolate reductase (DHFR) gene was found to be undermethylated only in its 5' promoter region; the remaining CCGG residues in the 30-kilobase (kb) DHFR gene were insensitive to digestion by HpaII. Each of 27 CpG residues that were part of an HpaII or HhaI cutting site within a 1.1-kb segment of the DHFR gene promoter region were found to be unmethylated. All 80 copies of the DHFR gene in methotrexate-resistant HeLa cell line exhibited this pattern of undermethylation of only the promoter region. This same region was shown to be DNase I hypersensitive in chromatin from normal cells and from those cells in which the DHFR gene was amplified. Again, all copies of the amplified gene exhibited DNase I hypersensitivity of the promoter region. The remainder of the 30-kb DHFR gene is both completely methylated and insensitive to DNase I digestion. Detailed mapping of the DNase I-hypersensitive region revealed four strong cutting sites within a 500-base pair segment immediately upstream from the DHFR coding sequence and a weak cutting site within intron I. Two of the strong DNase I cutting sites in chromatin were also sensitive to S1 nuclease nicking when this DNA fragment was part of supercoiled plasmid DNA. Promoter undermethylation and DNase I hypersensitivity, features previously shown for specialized and inducible genes, have now been shown to be characteristic of the constitutively expressed DHFR gene. That these features characterize all copies of the amplified DHFR gene in a methotrexate-resistant cell line suggest that all gene copies are transcriptionally active. PMID- 2982821 TI - Purification and characterization of a type II DNA topoisomerase from bovine calf thymus. AB - We report here the large scale purification of DNA topoisomerase II from calf thymus glands, using the unknotting of naturally knotted P4 phage DNA as an assay for enzymatic activity. Topoisomerase II was purified more than 1300-fold as compared to the whole cell homogenate, with 22% yield. Analysis of the purified enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed two bands of apparent molecular masses of 125 and 140 kDa. Tryptic maps of the two bands indicated that they derive from the same protein. Using these fragments, specific polyclonal antisera to topoisomerase II were raised in rabbits. Immunoblotting of whole cell lysates from various species indicated that topoisomerase II is well conserved among mammals and has a native subunit molecular mass of 180 kDa. Analytical sedimentation and gel filtration were used to determine a sedimentation coefficient of 9.8 S and a Stokes radius of 68 A. The calculated solution molecular mass of 277 kDa implies a dimer structure in solution. The purified topoisomerase II unknots P4 DNA in an ATP-dependent manner and is highly stimulated in its relaxation activity by ATP. A DNA-stimulated ATPase activity, as has been found with other type II topoisomerases, is associated with the purified enzyme. Approximate kinetic parameters for the ATPase reaction were determined to be: a Vmax of 0.06 nmol of ATP/(micrograms of protein) (min) and Km of 0.2 mM in the absence of DNA, and a Vmax of 0.2 nmol of ATP/(micrograms of protein) (min) and Km of 0.4 mM ATP in the presence of supercoiled plasmid DNA. PMID- 2982822 TI - Biochemical and immunological characterization of the secreted forms of human neutrophil gelatinase. AB - Human neutrophils contain a neutral metalloproteinase which degrades denatured collagens and potentiates the action of interstitial collagenase. This gelatinase is rapidly secreted from neutrophils stimulated with phorbol myristate acetate. The secreted enzyme has been purified by a combination of chromatography on DEAE cellulose and gelatin-Sepharose. The purified enzyme was latent and had a specific activity of 24,000 units. Estimated molecular weight obtained by gel filtration was 150,000-180,000. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme showed three bands with relative molecular weights of 225,000, 130,000, and 92,000. Electrophoresis in the presence of a reducing agent revealed a single band of Mr = 92,000. All the proteins seen on the unreduced gel were found to contain proteolytic activity against gelatin and native type V collagen. Polyclonal antibodies were prepared against the Mr = 130,000 and 92,000 proteins. When analyzed by immunoblotting, both antibodies recognized all three proteins. Furthermore, the identical three proteins were identified by the antibodies when crude culture medium was immunoblotted. The purified enzyme was inhibited by EDTA and 1,10-phenanthroline but not by serine or thiol proteinase inhibitors, suggesting that the enzyme is a metalloendoproteinase. The enzyme had little or no activity against common protein substrates such as bovine serum albumin or casein. Native type I collagen was not cleaved under conditions where native type V collagen was extensively degraded. PMID- 2982823 TI - The interaction of calmodulin with amphiphilic peptides. AB - Calmodulin has recently been shown to form exceptionally tight, calcium-dependent complexes with several natural peptides (Kdiss greater than 10(-7) M). These peptides were demonstrated to be capable of forming basic, amphiphilic alpha helices. To further illustrate the importance of this structural feature for calmodulin binding, several other amphiphilic alpha-helical peptides were tested for their ability to bind calmodulin. To monitor complexes of high affinity (greater than 10(8) M-1), a new competition assay was devised with Sepharose 4B conjugated melittin. Stoichiometries were assessed by electrophoresis and equilibrium size exclusion chromatography. Three peptides, which were designed to form idealized amphiphilic alpha-helices were tested. The basic peptides, N alpha 9-fluorenylmethoxycarboxyl-(FMOC)-(Leu-Lys-Lys-Leu-Leu-Lys-L eu)1 and FMOC-(Leu Lys-Lys-Leu-Leu-Lys-Leu)2 bind calmodulin in a 1:1 complex with dissociation constants of 150 and 3 nM, respectively. The acidic peptide, FMOC-(Leu-Glu-Glu Leu-Leu-Glu-Leu)2 failed to bind calmodulin, even at micromolar concentrations. Complex formation between calmodulin and the 14-residue basic peptide leads to an increase in the helicity of the complex which is attributed to an increase of about 50% in the helicity of the peptide. Calmodulin also interacts with the neutral alpha-helical peptide toxin delta-hemolysin. Concomitant with binding, the fluorescence maximum of the unique Trp residue increases 2-fold and is blue shifted. A dissociation constant could not be unambiguously estimated though, since delta-hemolysin has a strong tendency to self-aggregate. The above data support our hypothesis that a basic, amphiphilic alpha-helix is a structural feature which underlies the calmodulin-binding properties common to a variety of peptides. PMID- 2982824 TI - Amiloride directly inhibits growth factor receptor tyrosine kinase activity. AB - Addition of amiloride to A431 human epidermoid carcinoma cell membranes inhibited autophosphorylation of the epidermal growth factor (EGF) receptor. The tyrosine phosphorylation of histone H2B catalyzed by an affinity-purified preparation of EGF receptor was also inhibited by amiloride. The inhibition was noncompetitive with respect to histone but competitive with ATP, suggesting that amiloride may act as an ATP analogue which causes the formation of nonproductive enzyme substrate complexes. The tyrosine phosphorylation of histone H2B catalyzed by the purified EGF receptor was inhibited by amiloride at concentrations identical to those previously reported to block EGF action on cell proliferation (Ki = 350 microM). Amiloride similarly inhibited the tyrosine phosphorylation of the human placental insulin receptor and the platelet-derived growth factor receptor of Swiss 3T3 cells. Immunoprecipitation of the EGF receptor from A431 cells labeled for 24 h with [32P]phosphate demonstrated that amiloride decreased the phosphorylation of the EGF receptor on serine and threonine residues and blocked the effect of EGF to cause phosphorylation of the receptor on tyrosine residues. Phosphoamino acid analysis of total cell proteins indicated that amiloride inhibited the increase in phosphotyrosine levels caused by EGF. We conclude that amiloride directly inhibits the tyrosine kinase activity of the receptors for EGF, insulin, and platelet-derived growth factor in in vitro and can mediate such actions in vivo. This effect of amiloride demonstrates that it is unsuitable as a drug to test the hypothesis that the stimulation of the Na+/H+ antiporter is essential for mitogenic signaling by growth factor receptors. PMID- 2982825 TI - Fat cell adenylate cyclase system. Enhanced inhibition by adenosine and GTP in the hypothyroid rat. AB - Hypothyroidism is associated with an enhanced sensitivity of rat fat cells to the inhibitory action of adenosine and adenosine agonists. The sensitivity of the forskolin-stimulated cyclic AMP response of rat fat cells to the adenosine agonist N6-phenylisopropyladenosine is amplified 3-fold by hypothyroidism. Forskolin-stimulated adenylate cyclase activity is more sensitive to inhibition by this adenosine agonist in membranes of fat cells isolated from hypothyroid as compared to euthyroid rats. Hypothyroidism does not significantly alter the number of affinity of binding sites for N6-cyclohexyl[3H]adenosine or N6 phenylisopropyladenosine in membranes of rat fat cells. GTP-induced inhibition of forskolin-stimulated adenylate cyclase was markedly enhanced in the hypothyroid state, suggesting an alteration in the inhibitory regulatory component (Ni) mediated control of adenylate cyclase. Incubating membranes with [alpha-32P]NAD+ and preactivated pertussis toxin results in the radiolabeling of two peptides with Mr = 40,000 and 41,000 as visualized in autoradiograms of polyacrylamide gels run in sodium dodecyl sulfate. The amount of label incorporated by pertussis toxin into these two peptides (putative subunits of Ni) per mg of protein of membrane is increased 2-3-fold in the hypothyroid state. The amount of the stimulatory regulatory component, Ns, in fat cell membranes is not altered by hypothyroidism (Malbon, C. C., Graziano, M. P., and Johnson, G. L. (1984) J. Biol. Chem. 259, 3254-3260). The amplified response of hypothyroid rat fat cells to the inhibitory action of adenosine appears to reflect a specific increase in the activity and abundance of Ni. PMID- 2982826 TI - Adenine nucleotides directly stimulate pertussis toxin. AB - Both cholera toxin and pertussis toxin catalyzed ADP-ribosylation of purified bovine brain tubulin. The effect of cholera toxin was evident in the absence or presence of nucleotides. In contrast, pertussis toxin required adenine nucleotides for its ADP-ribosylating activity. ATP, ATP gamma S, App(NH)p, deoxy ATP, and ADP all supported pertussis toxin-catalyzed ADP-ribosylations in the absence or presence of EDTA, suggesting that nucleotide hydrolysis was not involved. Adenine nucleotides also promoted pertussis toxin-catalyzed ADP ribosylation of heat-treated bovine serum albumin. This result suggests that adenine nucleotides directly affect pertussis toxin. ATP stimulation of pertussis toxin-catalyzed hydrolysis of NAD to ADP-ribose supports this hypothesis. PMID- 2982827 TI - Regulation of neuroendocrine gene expression by alternative RNA processing. Colocalization of calcitonin and calcitonin gene-related peptide in thyroid C cells. AB - The generation of two discrete mRNAs encoding different polypeptide products in expression of the rat calcitonin, CGRP gene provides an example of developmentally determined alternative RNA processing events in the neuroendocrine system. Both mature mRNA transcripts and their peptide products, encoding either calcitonin or calcitonin gene-related peptide (CGRP) are co produced in identical thyroid C-cells, in a ratio of 95:1, respectively. These data suggest that the generation of the two mRNAs reflect the actions of a factor(s) dictating alternative RNA processing events. PMID- 2982828 TI - The ligninase of Phanerochaete chrysosporium generates cation radicals from methoxybenzenes. AB - The hemoprotein ligninase of Phanerochaete chrysosporium catalyzes, in the presence of H2O2, a variety of seemingly different oxidations in lignin and lignin model compounds. Here we show that the enzyme also catalyzes the oxidation of various methoxybenzenes. ESR spectroscopy shows that the compounds are oxidized to aryl cation radicals. These decompose, evidently by H2O addition. Thus, 1,4-dimethoxybenzene is converted to p-benzoquinone and methanol. We propose a unified mechanism, based on formation of aryl cation radicals, to explain the various reactions catalyzed by the ligninase. PMID- 2982829 TI - Cellular receptors for type beta transforming growth factor. Ligand binding and affinity labeling in human and rodent cell lines. AB - Type beta transforming growth factor (beta TGF) purified from human platelets to homogeneity as judged by NH2-terminal amino acid sequence analysis has been labeled with 125I to characterize its interaction with cellular receptors. Binding of 125I-beta TGF to target cells is temperature- and time-dependent, specific, saturable, and reversible. About 1.6-1.9 X 10(4) binding sites/cell with high affinity for beta TGF (Kd = 5.6-7.8 X 10(-11) M and 9.1-14 X 10(-11) M, respectively) are found in NRK-49F and BALB/c 3T3 cells. beta TGF receptors do not appear to undergo acute down-regulation by the ligand. Specific binding of 125I-beta TGF has been observed in several human, rat, and mouse fibroblast lines and in some, but not all, tumor-derived cell lines examined. 125I-beta TGF has been cross-linked to intact cells and isolated membrane preparations using disuccinimidyl suberate. Cells and isolated membranes from human, rat, and mouse origin affinity labeled with 125I-beta TGF exhibit a major labeled species of approximately 280 kilodaltons that has the properties of high affinity and specificity expected from a physiologically relevant beta TGF receptor. Minor labeled species of 70-90 kilodaltons are also labeled by 125I-beta TGF, but they correspond to molecular species with low apparent affinity (Kd approximately 10( 8) M) for 125I-beta TGF. PMID- 2982830 TI - Proteolytic conversion of [Met]enkephalin-Arg6-Gly7-Leu8 by brain synaptic membranes. Characterization of formed peptides and mechanism of proteolysis. AB - The degradation of the enkephalin-containing octapeptide Tyr-Gly-Gly-Phe-Met-Arg Gly-Leu (YGGFMRGL) was systematically investigated by incubating the peptide with synaptic membranes from rat striatum or with purified peptidases. The degradation products were derivatized with 4-dimethylamino-azobenzene-4'-isothiocyanate and then analyzed by high pressure liquid chromatography and by amino-terminal analysis. The incubation of YGGFMRGL with synaptic membranes yielded YGG, YGGF, YGGFM, and MR in a manner that was linear with respect to time. The corresponding carboxyl-terminal fragments FMRGL, MRGL, and RGL could not be detected, which suggests that the degradation of YGGFMRGL by synaptic membranes occurs by carboxypeptidase activity. The incubation of YGGFMRGL with different purified peptidases produced cleavage patterns unique from that seen with synaptic membranes. Enkephalinase recognized only the Gly-Phe bond to produce YGG and FMRGL. Thermolysin recognized the Gly-Phe bond and the Phe-Met bond to yield YGG, YGGF, FMRGL, and MRGL. Angiotensin-converting enzyme (ACE) produced primarily YGGF, MR, and lesser amounts of YGGFMR and YG. The formation of YGG, YGGF, and YGGFM by synaptic membranes could be stimulated 3-fold by the addition of 30 mM NaCl and inhibited by MK-422, an ACE inhibitor, with an IC50 of 3 nM. These data suggest that ACE, a dipeptidyl carboxypeptidase, is the primary enzyme involved in the degradation of YGGFMRGL in brain. ACE apparently works in concert with another carboxypeptidase in brain to yield YGGFM and YGG since the carboxyl terminal peptides RGL and FMRGL could not be detected. PMID- 2982831 TI - Inactivation of muscle adenylate kinase by site-specific destruction of tyrosine 95 using potassium ferrate. AB - Potassium ferrate, an analog of orthophosphate and a potent oxidizing agent, was found to irreversibly inactivate porcine muscle adenylate kinase. Inhibition was prevented by competitive inhibitors or substrates, indicating that the action of ferrate was site-specific. Inactivation was accompanied by the loss of Cys-25 and Tyr-95. P1,P5-di(adenosine 5')-pentaphosphate (10(-7) M), a powerful competitive inhibitor, gave 50% protection to the enzyme from ferrate inactivation. No loss of tyrosine or cysteine residues was observed under conditions of total protection. The degree of inactivation was proportional to the amount of Tyr-95 destroyed. However, Cys-25 was totally oxidized when only 55% inactivation had occurred. Partially inactivated enzyme exhibited a Km for ATP and AMP similar to that of the untreated enzyme. It appears that Cys-25 may be proximate to a phosphate-binding site but is not directly involved in the catalytic reaction. The results suggest that Tyr-95 is located in the vicinity of a phosphate-binding region of adenylate kinase and is essential for enzyme activity. PMID- 2982832 TI - Amplification and purification of the bacteriophage Mu encoded B transposition protein. AB - The A and B proteins encoded by the temperate bacteriophage Mu are involved in the high efficiency DNA transposition reaction which is the distinguishing feature of this phage. The genes encoding these early proteins were cloned in an expression vector under the control of the bacteriophage lambda leftward promoter. Under optimal conditions gpB was overproduced to account for 15% of the total cellular protein. The protein was purified to near homogeneity as determined by silver staining. Sequence analysis of the N terminus confirmed the identity of the purified protein as gpB. Proteolytic processing of the B protein does not occur at the amino terminus; the terminal methionine residue is quantitatively deformylated. The protein, which was found to be basic and a general DNA binding protein, was insoluble at low ionic strength in the absence, but not in the presence, of DNA. The B protein also displayed a tendency to aggregate at high ionic strength where it was soluble in the absence of DNA. In addition, the protein was characterized as to its amino acid composition and extinction coefficient at 280 nm. The purified protein is active in a soluble in vitro transposition-replication system. PMID- 2982833 TI - The ompA signal peptide directed secretion of Staphylococcal nuclease A by Escherichia coli. AB - The hybrid pre-enzyme formed by fusion of the signal peptide of the OmpA protein, a major outer membrane protein of Escherichia coli, to Staphylococcal nuclease A, a protein secreted by Staphylococcus aureus, is translocated across the cytoplasmic membrane of E. coli with concomitant cleavage of the signal peptide. A DNA fragment containing the coding sequence for the ompA signal peptide was initially ligated to a DNA fragment containing the coding sequence for nuclease A, with a linker sequence of 33 nucleotides separating the coding sequences. When this fused gene was induced, an enzymatically active nuclease was secreted into the periplasmic space; sequential Edman degradation of this protein revealed that the ompA signal peptide was removed at its normal cleavage site resulting in a modified version of the nuclease having 11 extra amino acid residues attached to the amino terminus of nuclease A. The 33 nucleotides between the coding sequences for the ompA signal peptide and the structural gene for nuclease A were subsequently deleted by synthetic oligonucleotide-directed site-specific mutagenesis. The nuclease produced by this hybrid gene was secreted into the periplasmic space and by sequential Edman degradation was identical to nuclease A. Thus, the ompA signal peptide is able to direct the secretion of fused staphylococcal nuclease A, and signal peptide processing occurs at the normal cleavage site. When the hybrid gene is expressed under the control of the lpp promoter, nuclease A is produced to the extent of 10% of the total cellular protein. PMID- 2982834 TI - Isolation and characterization of native adult osteonectin. AB - Noncollagenous bovine bone proteins were obtained from EDTA-solubilized extracts of adult bovine bone in the absence of denaturants. Native osteonectin was isolated from the noncollagenous bone proteins by ion-exchange chromatography using DEAE-Sephadex A-50 and DEAE-Sephadex A-25, followed by gel filtration on Sephadex G-100. Comparison of the physical and chemical properties (i.e. electrophoric mobility, amino acid composition and pI) of this protein with those reported by Termine et al. (Termine, J.D., Belcourt, A.B., Conn, K.M., and Kleinman, H.K. (1981) J. Biol. Chem. 256, 10403-10408) indicate that this protein is osteonectin. Sedimentation equilibrium analyses in the presence of 6 M guandinium chloride, 10 mM Ca2+, 10 mM EDTA, or 0.15 M NaCl all yielded a molecular weight of 29,100 +/- 900. 125I-Osteonectin underwent saturable and exchangeable binding to hydroxyapatite and calf skin collagen. Eleven mg of 125I osteonectin bound to 1.0 g of hydroxyapatite with a Kd of 8 X 10(-8) M. The intrinsic fluorescence of bovine osteonectin was partially quenched when micromolar Ca2+ was added, indicating a high affinity Ca2+ interaction. Native osteonectin was found to reduce the rate of hydroxyapatite crystal seeded growth by 50% (1 IU) at a concentration of 1.6 X 10(-7) M at pH 7.4, 37 degrees C in 0.15 M NaCl. This makes osteonectin one of the most potent inhibitors of hydroxyapatite formation presently known and more than 5 times as effective as bone Gla protein (1 IU = 8 X 10(-7) M). PMID- 2982835 TI - Substrate specificity of beta-collagenase from Clostridium histolyticum. AB - The substrate specificity of beta-collagenase from Clostridium histolyticum has been investigated by measuring the rate of hydrolysis of more than 50 tri-, tetra , penta-, and hexapeptides covering the P3 to P3' subsites of the substrate. The choice of peptides was patterned after sequences found in the alpha 1 and alpha 2 chains of type I collagen. Each peptide contained either a 2-furanacryloyl (FA) or cinnamoyl (CN) group in subsite P2 or the 4-nitrophenylalanine (Nph) residue in subsite P1. Hydrolysis of the P1-P1' bond produces an absorbance change in these chromophoric peptides that has been used to quantitate the rates of their hydrolysis under first order conditions ([S] much less than KM) from kcat/KM values have been obtained. The identity of the amino acids in all six subsites (P3-P3') markedly influences the hydrolysis rates. In general, the best substrates have Gly in subsites P3 and P1', Pro or Ala in subsite P2', and Hyp, Arg, or Ala in subsite P3'. This corresponds well with the frequency of occurrence of these residues in the Gly-X-Y triplets of collagen. In contrast, the most rapidly hydrolyzed substrates do not have residues from collagen-like sequences in subsites P2 and P1. For example, CN-Nph-Gly-Pro-Ala is the best known substrate for beta-collagenase with a kcat/KM value of 4.4 X 10(7) M-1 min 1, in spite of the fact that there is neither Pro nor Ala in P2 or Hyp nor Ala in P1. These results indicate that the previously established rules for the substrate specificity of the enzyme require modification. PMID- 2982836 TI - EPR studies of iron-sulfur clusters in isolated subunits and subfractions of NADH ubiquinone oxidoreductase. AB - The thermodynamic and EPR characteristics of the iron-sulfur clusters of NADH ubiquinone oxidoreductase have been examined in various subfractions and subunits of the enzyme. These were obtained by fragmentation of the enzyme with chaotropic agents and detergent and salt fractionation. We provide evidence for the presence of three tetranuclear clusters and five or six binuclear clusters, accounting well for the chemically determined iron content of this enzyme (22-24 atoms/molecule of FMN). Some of the clusters can be identified with EPR detectable species in intact NADH-ubiquinone oxidoreductase and, by combining information on subunit topography and spin-spin interactions between redox centers in the native enzyme, we propose a tentative scheme for the spatial organization of these iron-sulfur clusters in the enzyme and in the membrane. PMID- 2982837 TI - Beta-epidermal growth factor is the des-asparaginyl form of the polypeptide. AB - Reversed-phase high performance liquid chromatography of mouse epidermal growth factor (EGF) yielded two major forms, alpha- and beta-EGF, and a minor component, gamma-EGF. All three forms exhibited receptor-binding activity. Analysis of native alpha- and beta-EGF by mass spectrometry and partial Edman degradation led us to propose that alpha-EGF has a primary structure equivalent to that originally reported for EGF and that beta-EGF is the des-asparaginyl form of the polypeptide. When the purified alpha- and beta-polypeptides were cultured with human embryonic palatal mesenchymal cells stimulation of cell proliferation was observed at concentrations as low as 0.01 ng/ml with maximal stimulation occurring at about 1 ng/ml. Essentially no difference was noted in the mitogenic potency of the two forms. This suggests that the NH2-terminal region of EGF is not critical for mitogenic activity. PMID- 2982838 TI - Nuclear magnetic resonance studies of fructose 2,6-bisphosphate and adenosine 5' monophosphate interaction with bovine liver fructose-1,6-biphosphatase. AB - 1H and 31P nuclear magnetic resonance was used to investigate the interaction of AMP and fructose 2,6-bisphosphate (Fru-2,6-P2) with bovine liver fructose-1,6 bisphosphatase. Mn2+ bound to fructose-1,6-bisphosphatase was used as a paramagnetic probe to map the active and AMP allosteric sites of fructose-1,6 bisphosphatase. Distances between enzyme-bound Mn2+ and the phosphorus atoms at C 6 of fructose-6-P and alpha-methyl-D-fructofuranoside 1,6-bisphosphate were identical, and the enzyme-Mn to phosphorus distance determined for the C-6 phosphorus atom of Fru-2,6-P2 was very similar to these values. Likewise, the enzyme-Mn to phosphorus distances for Pi, the C-1 phosphorus atom of alpha-methyl D-fructofuranoside 1,6-bisphosphate, and the C-2 phosphorus atom of Fru-2,6-P2 agreed within 0.5 A. The distance between enzyme-bound Mn2+ and the phosphorus atom of AMP was significantly shorter than the distances obtained for any of the aforementioned ligands, but the presence of Fru-2,6-P2 caused the enzyme-Mn to phosphorus distance for AMP to lengthen markedly. NMR line broadening of AMP protons was studied at various temperatures. The dissociation rate constant was found to be greater than 20 s-1. It was concluded that Fru-2,6-P2 strongly affects the interaction of AMP with fructose-1,6-bisphosphatase and that the sugar most likely acts at the active site of the enzyme. PMID- 2982839 TI - Inhibition of hepatic alpha 1-adrenergic effects and binding by phorbol myristate acetate. AB - Treatment of isolated hepatocytes with the tumor-promoting agent, 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA) produced a time- and dose-dependent, non competitive inhibition of alpha 1-adrenergic responses, including the activation of phosphorylase, increase in Ca2+ efflux, increase in free cytosolic Ca2+, and release of myo-inositol-1,4,5-P3. The actions of [8-arginine] vasopressin (AVP) on liver cells were also inhibited by PMA, but the inhibition could be overcome by high AVP concentrations. No significant inhibition of beta-adrenergic and glucagon-mediated activation of phosphorylase was induced by PMA and no inhibitory or synergistic effects of PMA were observed on the dose-dependent activation of phosphorylase by the Ca2+ ionophore A23187. In radioligand binding studies, PMA did not directly interfere with [3H]prazosin specific binding, the displacement of [3H]prazosin by (-)-norepinephrine nor with [3H]AVP specific binding to purified liver plasma membranes. Plasma membranes prepared from livers perfused with PMA exhibited a 30-44% reduction in [3H]prazosin binding capacity. Under identical conditions [3H]AVP binding was unchanged. The alpha 1-receptors remaining in membranes from PMA-treated livers had equivalent affinities for [3H]prazosin and (-)-norepinephrine, and were unaffected in terms of coupling to guanine nucleotide-regulating proteins as indicated by the ability of guanosine 5'-(beta, gamma-imido)triphosphate to promote the conversion of the remaining alpha 1-receptors into a low affinity state. These data indicate that tumor promoters are potent antagonists of alpha 1-adrenergic and vasopressin (low dose) responses in liver. It is proposed that PMA acting via protein kinase C (which presumably mediates the action of PMA) exerts its inhibitory action on alpha 1 adrenergic responses at the alpha 1-adrenergic receptor itself and also at a site close to or before myo-inositol-1,4,5-P3 release. PMID- 2982840 TI - Activation of human platelets by N-substituted aminophospholipids. AB - N-(7-nitro-2,1,3-benzoxadiazol-4-yl) phosphatidylserine (NBD-PS), a fluorescent phospholipid synthesized from phosphatidylserine by reaction with NBD-chloride, caused platelet shape change and aggregation when added at micromolar concentrations to suspensions of washed human platelets in the absence of added fibrinogen. Platelet aggregation by NBD-PS was accompanied by thromboxane synthesis and secretion of contents from dense, alpha-, and lysosomal granules in the absence of appreciable platelet damage. Indomethacin completely inhibited NBD PS-induced thromboxane synthesis, but platelet aggregation and [14C]serotonin secretion were only slightly inhibited. Neither inhibition of the ADP-dependent pathway with creatine phosphate/creatine kinase plus ATP, alone or in combination with indomethacin, nor maximum elevation of cyclic AMP by treatment with prostaglandin I2 and theophylline completely inhibited NBD-PS-induced platelet aggregation or [14C]serotonin secretion. Platelet effects of NBD-PS were specific in that neither phosphatidylserine nor lyso-NBD-PS were similarly active. The activation of platelets by NBD-PS is not attributable to the NBD moiety exclusively since acylation of the amino group with 5-dimethylaminonaphthalene-1 sulfonyl-chloride yielded a similarly active derivative. Dansylated phosphatidylethanolamine was also active. The findings indicate that NBD-PS and other N-substituted aminophospholipids can activate a central pathway of platelet secretion and aggregation that is independent of released ADP and thromboxane formation and is only partially controlled by platelet cyclic AMP. PMID- 2982841 TI - The identification of histidine ligands to cytochrome a in cytochrome c oxidase. AB - A histidine auxotroph of Saccharomyces cerevisiae has been used to metabolically incorporate [1,3-15N2] histidine into yeast cytochrome c oxidase. Electron nuclear double resonance (ENDOR) spectroscopy of cytochrome a in the [15N]histidine-substituted enzyme reveals an ENDOR signal which can be assigned to hyperfine coupling of a histidine 15N with the low-spin heme, thereby unambiguously identifying histidine as an axial ligand to this cytochrome. Comparison of this result with similar ENDOR data obtained on two 15N-substituted bisimidazole model compounds, metmyoglobin-[15N]imidazole and bis[15N]imidazole tetraphenyl porphyrin, provides strong evidence for bisimidazole coordination in cytochrome a. PMID- 2982842 TI - N-(2-Aminoethyl)-5-isoquinolinesulfonamide, a newly synthesized protein kinase inhibitor, functions as a ligand in affinity chromatography. Purification of Ca2+ activated, phospholipid-dependent and other protein kinases. AB - We designed a simple procedure for the purification of Ca2+-activated, phospholipid-dependent protein kinase (protein kinase C) from rabbit brain, using affinity chromatography with a new affinity adsorbent. The adsorbent was synthesized by attaching the amino residue of N-(2-aminoethyl)-5 isoquinolinesulfonamide (H-9) to cyanogen bromide-activated Sepharose. H-9 is a potent competitive inhibitor of protein kinase C, cGMP-, and cAMP-dependent protein kinase with respect to ATP and exhibits inhibition constants of 18, 0.87, and 1.9 microM, respectively (Hidaka, H., Inagaki, M., Kawamoto, S., and Sasaki, Y. (1984) Biochemistry, 23, 5036). A 960-fold purification was achieved in the two-step procedure, which entailed DEAE-cellulose and the affinity chromatography. The resultant preparation was essentially homogeneous, as indicated by polyacrylamide gel electrophoresis under conditions of denaturation with sodium dodecyl sulfate. The affinity of protein kinase C for the H-9 Sepharose was high, and the enzyme could not be eluted either by a high concentration of sodium chloride or by 40% glycerol. The protein kinase C could be eluted from H-9-Sepharose by the buffer containing both 0.2 M NaCl and 20% glycerol, thereby suggesting that the binding between protein kinase C and H-9 Sepharose was due to both hydrophobic and electrostatic interactions. H-9 coupled to Sepharose retained both cyclic nucleotide-dependent protein kinases and protein kinase C, and these enzymes could be eluted separately by the buffer containing L-arginine, a potent inhibitor of these three kinases. The novel aspects of these three multifunctional protein kinases can thus be investigated using isoquinolinesulfonamide derivatives. PMID- 2982843 TI - Iron deposition in apoferritin. Evidence for the formation of a mixed valence binuclear iron complex. AB - A preliminary EPR investigation of iron accumulation in apoferritin has identified paramagnetic species generated during the early stage of iron deposition within the apoprotein shell. A featureless resonance at g' = 4.3, attributable to solitary high spin Fe3+ ions bound to the protein, is generated when Fe(II) is added to apoferritin at a level of 0.5 Fe/subunit (12 Fe/molecule) followed by air oxidation. This resonance accounts for 36% of the added iron. The remainder is EPR-silent and is probably present as oligomeric Fe3+ species. The intensity of the g' = 4.3 signal is reduced 3-fold upon anaerobic addition of 5 Fe(II)/subunit as a new iron resonance with g' values of 1.94, 1.87, and 1.80 is generated. This signal is observable only at temperatures near that of liquid helium and resists saturation at power levels of 100 milliwatts. Its distinctive g-factors, temperature dependence, and saturation characteristics suggest that it arises from a spin-coupled Fe(II)-Fe(III) dimer having a net electron spin of 1/2. In accord with this idea, the signal disappears when air is admitted, presumably because of oxidation of the Fe(II). The proposed mixed valence dimer may be an important intermediate formed during the initiation of core formation within the protein shell. PMID- 2982844 TI - Mouse ornithine decarboxylase. Complete amino acid sequence deduced from cDNA. AB - cDNA containing the full coding region of mouse ornithine decarboxylase was isolated. The complete nucleotide sequence of the cDNA was determined by the dideoxy method, and the amino acid sequence of ornithine decarboxylase was thereby deduced. The protein contains 461 amino acids and has a molecular weight of 51,172. The isoelectric point is predicted from the deduced amino acid sequence to be 5.1. On the basis of its amino acid sequence, the protein is predicted to be comprised predominantly of alternating domains of alpha-helix and beta-sheet. PMID- 2982845 TI - Inhibition of rabbit lung angiotensin-converting enzyme by N alpha-[(S)-1-carboxy 3-phenylpropyl]L-alanyl-L-proline and N alpha-[(S)-1-carboxy-3-phenylpropyl]L lysyl-L-proline. AB - Two novel peptide analogs, N alpha-[(S)-1-carboxy-3-phenylpropyl]L-alanyl-L proline and the corresponding L-lysyl-L-proline derivative, have been demonstrated to be potent competitive inhibitors of purified rabbit lung angiotensin-converting enzyme: Ki = 2 and 1 X 10(-10) M, respectively, at pH 7.5, 25 degrees C, and 0.3 M chloride ion. Second-order rate constants for addition of these inhibitors to enzyme under the same conditions are in the range 1-2 X 10(6) M-1 s-1; first-order rate constants for dissociation of the EI complexes are in the range 1-4 X 10(-4) s-1. The association rate constants are similar to those measured for D-3-mercapto-2-methylpropanoyl-L-proline, captopril, but the dissociation rate constants are severalfold slower and account for the higher affinity of these inhibitors for the enzyme. The dissociation constant for the EI complex containing N alpha-[(S)-1-carboxy-3-phenylpropyl]L-alanyl-L-proline is pH dependent, and reaches a minimum at approximately pH 6: Ki = 4 +/- 1 X 10(-11) M. The pH dependence is consistent either with a model for which the protonation state of the secondary nitrogen atom in the inhibitor determines binding affinity, or one for which ionizations on the enzyme alone influence affinity for these inhibitors. The affinity of this inhibitor for the zinc-free apoenzyme is 2 X 10(4) times less than for the zinc-free apoenzyme is 2 X 10(4) times less than that for the holoenzyme. If considered as a "collected product" inhibitor, N alpha-[(S)-1-carboxy-3-phenylpropyl]L-alanyl-L-proline appears to derive an additional factor of 375 M in its affinity for the enzyme compared to that of the two products of its hypothetical hydrolysis, a consequence of favorable entropy effects. PMID- 2982846 TI - Purification of angiotensin-converting enzyme from rabbit lung and human plasma by affinity chromatography. AB - Lisinopril (N alpha-[(S)-1-carboxy-3-phenylpropyl]L-lysyl-L-proline), a potent angiotensin-converting enzyme inhibitor, is an exceptionally selective affinity chromatography ligand for this enzyme. Affinity chromatography furnishes electrophoretically homogeneous enzyme directly from crude homogenates of rabbit lung tissue, a 1,000-fold purification; also, it affords a 100,000-fold enrichment of the more rare human plasma enzyme in a single step. The affinity of angiotensin-converting enzyme for the Sepharose-spacer-lisinopril matrix (Ki matrix = 1 X 10(-5) M) is weak compared to its affinity for free lisinopril (Ki = 1 X 10(-10) M). The capacity of the affinity column is described quantitatively as a function of Ki matrix, lisinopril, and enzyme concentrations. The recovery of bound enzyme is low in chromatography of crude tissue samples (10-40%), although it approaches a reversible process (70-100%) with pure enzyme. The holoenzyme is converted to Zn2+-free apoenzyme to effect removal of lisinopril. In this process, the rate constant for spontaneous dissociation of Zn2+ from free enzyme is 1 X 10(-2) s-1 (t 1/2 = 1 min), which places a lower limit of 3 X 10( 10) M on the dissociation constant of Zn2+ at neutral pH from angiotensin converting enzyme. The exceptional selectivity of lisinopril as an affinity chromatography ligand for angiotensin-converting enzyme suggests it is among the most specific inhibitors designed for any enzyme. PMID- 2982847 TI - Transcription of the Escherichia coli adenylate cyclase gene is negatively regulated by cAMP-cAMP receptor protein. AB - The regulatory region of the Escherichia coli cya gene was analyzed by using S1 nuclease mapping and in vitro transcription experiments. The cya gene was transcribed, both in vivo and in vitro, from one major promoter (P2) and two weak promoters (P1 and P1') that are located about 200 base pairs upstream of P2. The transcription from P2 was specifically inhibited by cAMP-CRP (cAMP receptor protein) in vitro. This regulatory mechanism was shown to be physiologically relevant through quantitative analyses of the cya mRNA in intact cells by S1 and dot blot assays. DNase I protection experiments revealed that cAMP-CRP binds to the cya DNA region between +11 and -20, in which a consensus CRP binding sequence is present. Moreover, it was found that cAMP-CRP alters the binding of RNA polymerase to the promoter region, thus inhibiting the transcription of the cya gene. PMID- 2982848 TI - Biosynthesis of tissue inhibitor of metalloproteinases by human fibroblasts in culture. Stimulation by 12-O-tetradecanoylphorbol 13-acetate and interleukin 1 in parallel with collagenase. AB - Biosynthesis of the glycoprotein tissue inhibitor of metalloproteinases (TIMP) by human fibroblasts in culture has been characterized by functional assays, immunoprecipitation, and immunocytochemistry with a monospecific antiserum. As determined by radiolabeling with [35S]methionine, immunoprecipitation, and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the secreted form of TIMP had an Mr of 29,000, whereas the form associated with the cell layer had an Mr of 24,000. Unstimulated human lung fibroblasts (HFL-1) secreted TIMP at the rate of approximately 2 micrograms/10(6) cells/24 h, and normal foreskin fibroblasts (HS 27) and skin fibroblasts from a patient with Hurler's disease (GM 1391) secreted TIMP at 0.3 and 0.2 micrograms/10(6) cells/24 h, respectively. Secretion of TIMP was stimulated up to 10-fold by treating the cells with 20-100 ng/ml of 12-O-tetradecanoylphorbol 13-acetate or 10 units/ml of human interleukin 1. In the stimulated HFL-1 cells, TIMP accounted for 0.03-0.09% of the total [35S]methionine incorporated into protein, and 0.3-0.8% of the [35S]methionine in secreted protein. Although TIMP accounted for a relatively small proportion of total protein synthesis of the fibroblasts, greater than 80% of untreated and greater than 95% of stimulated fibroblasts synthesized TIMP, as determined by indirect immunofluorescence. The treatments of the human fibroblasts that increased TIMP secretion also induced synthesis and secretion of proenzyme forms of collagenase, indicating that degradative enzymes and their controlling inhibitors may be synthesized in parallel under certain conditions. PMID- 2982849 TI - Cloning, nucleotide sequence, and expression of one of two genes coding for yeast elongation factor 1 alpha. AB - One gene coding for yeast cytoplasmic elongation factor 1 alpha (EF-1 alpha) was isolated by colony hybridization using a cDNA probe prepared from purified EF-1 alpha mRNA. A recombinant plasmid, pLB1, with a 6-kilobase yeast DNA insert, was found by hybrid selection and translation experiments to carry the entire gene. The nucleotide sequence of the gene with its 5'- and 3'-flanking regions was determined. The 5' and 3' ends of EF-1 alpha mRNA were localized by the S1 nuclease mapping technique. The cloned gene, called TEF1, encodes a protein of 458 amino acids (Mr = 50,071) in a single, uninterrupted reading frame. The amino acid sequence shows a strong homology with several domains of Artemia salina EF-1 alpha cytoplasmic factor, as evidenced by diagonal dot matrix analysis. Protein sequence homology is comparatively much lower with the yeast mitochondrial elongation factor. S1 nuclease mapping of the mRNA, hybridization analysis of chromosomal DNA using intragenic or extragenic DNA probes, and gene disruption experiments demonstrated the existence of two genes coding for the cytoplasmic elongation factor EF-1 alpha/haploid genome. The presence of an intact chromosomal TEF1 gene is not essential for growth of haploid yeast cells. PMID- 2982850 TI - Kinetics of dephosphorylation of eIF-2(alpha P) and reutilization of mRNA. AB - Phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF-2) causes mRNA to accumulate in 48 S complexes containing Met-tRNAf and eIF-2(alpha P). When the eIF-2 alpha kinase is inhibited by 2-aminopurine, the mRNA is slowly transferred from 48 to 80 S initiation complexes after an initial lag. The cause of this lag was examined by investigating whether mRNA and Met-tRNAf dissociated from 48 S complexes before binding to 80 S. Both compounds were quantitatively transferred from 48 to 80 S complexes after addition of 2-aminopurine and the eIF 2(alpha P) bound to 48 S complexes was dephosphorylated after an initial lag more slowly than unbound eIF-2(alpha P), which was rapidly dephosphorylated. the eIF 2(alpha P) in isolated 48 S complexes was slowly dephosphorylated by partially purified lysate phosphatases, whereas free eIF-2(alpha P) was readily dephosphorylated. These results indicated that 48 S complexes could directly join to a 60 S ribosomal subunit after eIF-2(alpha P) dephosphorylation. The lag and slow kinetics of dephosphorylation of eIF-2(alpha P) bound to 48 S complexes accounted for the slow transfer of mRNA from 48 to 80 S complexes. Moreover, the mRNA bound to 48 S complexes was more susceptible to cleavage by an endonuclease than mRNA in polyribosomes, as shown by activating the (2'-5')oligo(A)-dependent endonuclease. This finding is discussed in view of the possible role of eIF-2 alpha kinase and endonuclease in the inhibition of viral mRNA translation in interferon-treated cells. PMID- 2982851 TI - Initiation of DNA replication at the primary origin of bacteriophage T7 by purified proteins. Site and direction of initial DNA synthesis. AB - In vivo, T7 DNA replication is initiated 15% of the distance from the genetic left end of the chromosome. This site, the primary origin of replication, consists of a 200-base pair (bp) intergenic segment from 14.5 to 15.0% within which are located two tandem T7 RNA polymerase promoters (phi 1.1A and phi 1.1B) followed by a 61-bp AT-rich (79% A + T) region. A fragment of T7 DNA containing the primary origin has been inserted into plasmids in order to facilitate studies on initiation in vitro. Initiation of DNA synthesis can be reconstituted using T7 RNA polymerase, T7 DNA polymerase, and T7 origin-containing plasmid DNAs. DNA synthesis is stimulated greatly by the T7 gene 4 protein, an enzyme that has helicase and primase activities. When T7 gene 4 protein is present, replication primarily yields partially replicated Y-form molecules as observed by electron microscopy. Synthesis is unidirectional and the branches of the Y-form molecules are uniform in size, with the branch point of the Y located at the origin. Using restriction enzyme analysis, DNA synthesis has been shown to proceed in the same direction (rightward with respect to the T7 genetic map) as transcription from the two promoters located at the origin. Initiation of DNA synthesis in the opposite direction requires the addition of a single-stranded DNA-binding protein (Fuller, C.W., and Richardson, C.C. (1985) J. Biol. Chem. 260, 3197-3206). The initial products of DNA synthesis have been analyzed by polyacrylamide gel electrophoresis. These DNAs have 10 to 60 ribonucleotides covalently linked to their 5' termini. These RNA primers arise by transcription from each of the two promoters, phi 1.1A and phi 1.1B, located within the primary origin. PMID- 2982852 TI - Evidence for N coordination to Fe in the [2Fe-2S] clusters of Thermus Rieske protein and phthalate dioxygenase from Pseudomonas. AB - Rieske-type iron/sulfur proteins and several NADH-dependent oxygenases contain Fe/S clusters with similar spectral and magnetic properties. Purified Rieske iron/sulfur protein from Thermus thermophilus contains two apparently identical [2Fe-2S] clusters in a polypeptide having only four cysteine residues, and it has been proposed that each Fe/S cluster is coordinated to two cysteine S-atoms and to an unknown number of other non-sulfur atoms (Fee, J. A., Findling, K. L., Yoshida, T., Hille, R., Tarr, G. E., Hearshen, D. O., Dunham, W. R., Day, E. P., Kent, T. A., and Munck, E. (1984) J. Biol. Chem. 259, 124-133). We have examined the Rieske protein from Thermus and the phthalate dioxygenase from Pseudomonas cepacia with electron nuclear double resonance (ENDOR) and pulsed EPR methods and report here evidence for the direct coordination of nitrogenous ligands to the Fe/S clusters in these proteins. The electron nuclear double resonance signals arising from 14N have been interpreted in terms of a strongly coupled ligand with AN = approximately 26-28 MHz and a weakly coupled ligand with AN = approximately 9 MHz. The pulsed EPR spectrum shows a rich pattern of lines in the Fourier transformed data having peaks in the range of 0.8 to 6.7 MHz. The lower frequency resonances are tentatively associated with coupling of the unpaired spin to the remote N-atoms of coordinated imidazole rings. PMID- 2982853 TI - Increased cyclic GMP levels lead to a stimulation of elastin production in ligament fibroblasts that is reversed by cyclic AMP. AB - The effects of cyclic nucleotides on elastin synthesis were studied in ligamentum nuchae fibroblasts by adding exogenous cyclic nucleotide derivatives or beta adrenergic agents to cell culture medium. Elastin synthesis was enhanced (approximately 80%) by dibutyryl cGMP (Bt2cGMP) in concentrations ranging from 0.01 to 100 nM. Two other cGMP derivatives, 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br-cGMP) and 2'-deoxy-cGMP, were also potent stimulators of elastin synthesis. In the absence of calcium, basal elastin production was substantially decreased (40% of control) and cGMP analogs no longer stimulated elastin synthesis, suggesting a role for calcium in the cGMP response. Bt2cAMP had no demonstrable effect on elastin production except at high concentrations which produced a nonspecific decrease equivalent to the decrease in total protein synthesis. Similarly, elevation of endogenous cellular cAMP levels by beta adrenergic stimulation produced no change in elastin production. When 8-Br-cGMP was added to cells together with Bt2cAMP, cGMP-dependent stimulation of elastin production was abolished by cAMP in a dose-dependent fashion. These results suggest a coordinated means by which elastin production is controlled in ligament cells, i.e. increased cGMP levels lead to a stimulation of elastin production that is reversed by cAMP. PMID- 2982854 TI - Ferritin and superoxide-dependent lipid peroxidation. AB - Ferritin was found to promote the peroxidation of phospholipid liposomes, as evidenced by malondialdehyde formation, when incubated with xanthine oxidase, xanthine, and ADP. Activity was inhibited by superoxide dismutase but markedly stimulated by the addition of catalase. Xanthine oxidase-dependent iron release from ferritin, measured spectrophotometrically using the ferrous iron chelator 2,2'-dipyridyl, was also inhibited by superoxide dismutase, suggesting that superoxide can mediate the reductive release of iron from ferritin. Potassium superoxide in crown ether also promoted superoxide dismutase-inhibitable release of iron from ferritin. Catalase had little effect on the rate of iron release from ferritin; thus hydrogen peroxide appears to inhibit lipid peroxidation by preventing the formation of an initiating species rather than by inhibiting iron release from ferritin. EPR spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide was used to observe free radical production in this system. Addition of ferritin to the xanthine oxidase system resulted in loss of the superoxide spin trap adduct suggesting an interaction between superoxide and ferritin. The resultant spectrum was that of a hydroxyl radical spin trap adduct which was abolished by the addition of catalase. These data suggest that ferritin may function in vivo as a source of iron for promotion of superoxide-dependent lipid peroxidation. Stimulation of lipid peroxidation but inhibition of hydroxyl radical formation by catalase suggests that, in this system, initiation is not via an iron-catalyzed Haber-Weiss reaction. PMID- 2982856 TI - (Na,K)-ATPase and Ouabain binding in reticulocytes from dogs with high K and low K erythrocytes and their changes during maturation. AB - The present study demonstrated that dog reticulocytes had considerable amounts of (Na,K)-ATPase, but lost it rapidly during maturation into erythrocytes. Furthermore, reticulocytes from dogs possessing erythrocytes characterized with high (Na,K)-ATPase activity and high K, low Na concentrations (HK dogs; Maede, Y., Inaba, M., and Taniguchi, N. (1983) Blood 61,493-499) had more ouabain binding sites than cells from normal dogs (LK dogs). Our results were as follows: i) The maximal binding capacities (Bmax) for ouabain binding at equilibrium were approximately 0 and 1,500 binding sites/cell in LK and HK dog erythrocytes, respectively. ii) Reticulocytes from LK dogs possess approximately 5,700 ouabain binding sites/cell. iii) The Bmax value for ouabain in HK reticulocytes was about 10,000 sites/cell, being 2-fold that in LK reticulocytes. iv) Ouabain-sensitive fluxes of 24Na and 42K in each type of reticulocyte were compatible with the number of ouabain binding sites on the cells. v) Ouabain binding capacity, as well as (Na,K)-ATPase activity, in the reticulocytes from LK dogs fell rapidly to nearly zero during the maturation into erythrocytes. vi) Although reticulocytes from HK dogs also showed a similar regression of (Na,K)-ATPase during maturation, they retained a certain number of ouabain binding sites even after maturation, resulting in the high activity of (Na,K)-ATPase in HK erythrocyte membrane. PMID- 2982855 TI - Photosynthetic membrane development in Rhodopseudomonas sphaeroides. Spectral and kinetic characterization of redox components of light-driven electron flow in apparent photosynthetic membrane growth initiation sites. AB - The kinetics of light-driven electron flow and the nature of redox centers at apparent photosynthetic membrane growth initiation sites in Rhodopseudomans sphaeroides were compared to those of intracytoplasmic photosynthetic membranes. In sucrose gradients, these membrane growth sites sediment more slowly than intracytoplasmic membrane-derived chromatophores and form an upper pigmented band. Cytochromes c1, c2, b561, and b566 were demonstrated in the upper fraction by redox potentiometry; c-type cytochromes were also detected electrophoretically. Signals characteristic of light-induced reaction center bacteriochlorophyll triplet and photooxidized reaction center bacteriochlorophyll dimer states were observed by EPR spectroscopy but the Rieske iron-sulfur signal of the ubiquinol-cytochrome c2 oxidoreductase was present at a 3-fold reduced level on a reaction center basis in comparison to chromatophores. Flash-induced absorbance measurements of the upper pigmented fraction demonstrated reaction center primary and secondary semiquinone anion acceptor signals, but cytochrome b561 photoreduction and cytochrome c1/c2 reactions occurred at slow rates. This fraction was enriched approximately 2- and 4-fold in total b- and c-type cytochromes, respectively, per reaction center over chromatophores, but photoreducible b-type cytochrome was lower. Measurements of respiratory activity indicated a 1.6-fold higher level of succinate-cytochrome c oxidoreductase/reaction center than in chromatophores, but the apparent turnover rates in both preparations were low. Overall, the results suggest that complete cycles of rapid, light-driven electron flow do not occur merely by introduction of newly synthesized reaction centers into respiratory membrane, but that subsequent synthesis and assembly of appropriate components of the ubiquinol cytochrome c2 oxidoreductase is required. PMID- 2982857 TI - Identification of a trpG-related glutamine amide transfer domain in Escherichia coli GMP synthetase. AB - An improved method was developed to align related protein sequences and search for homology. A glutamine amide transfer domain was identified in an NH2-terminal segment of GMP synthetase from Escherichia coli. Amino acid residues 1-198 in GMP synthetase are homologous with the glutamine amide transfer domain in trpG X D encoded anthranilate synthase component II-anthranilate phosphoribosyltransferase and the related pabA-encoded p-aminobenzoate synthase component II. This result supports a model for gene fusion in which a trpG-related glutamine amide transfer domain was recruited to augment the function of a primitive NH3-dependent GMP synthetase. Sequence analyses emphasize that glutamine amide transfer domains are thus far found only at the NH2 terminus of fused proteins. Two rules are formulated to explain trpG and trpG-related fusions. (i) trpG and trpG-related genes must have translocated immediately up-stream of genes destined for fusion in order to position a glutamine amide transfer domain at the NH2 terminus after fusion. (ii) trpG and trpG-related genes could not translocate adjacent to a regulatory region at the 5' end of an operon. These rules explain known trpG-like fusions and explain why trpG and pabA are not fused to trpE and pabB, respectively. Alignment searches of GMP synthetase with two other enzymes that bind GMP, E. coli amidophosphoribosyltransferase and human hypoxanthine-guanine phosphoribosyltransferase, suggest a structurally homologous segment which may constitute a GMP binding site. PMID- 2982859 TI - The separate estimation of cAMP intracellularly bound to the regulatory subunits of protein kinase I and II in glucagon-stimulated rat hepatocytes. AB - A method is described for the separate determination of cAMP intracellularly bound to the regulatory moieties (RI and RII) of protein kinase I and II. The cAMP endogenously bound to RI or RII in hepatocyte extract was adsorbed to protein A-agarose beads coated with antibodies against RI or RII. The endogenously bound cAMP was eluted from the washed beads with dilute acetic acid before being assayed. By all criteria tested, the present method did not perturb the intracellularly established equilibrium between bound and free cAMP. Stabilization of R X cAMP complexes was achieved by including sulfate in the extraction medium and sulfate/glycerol during the subsequent steps. Hepatocytes were isolated from fed male rats and contained about 0.25 pmol of RI and 0.2 pmol of RII per 10(5) cells. An intracellular titration of the cAMP binding sites of RI and RII was achieved by incubating the cells with various concentrations (1 pM to 10 nM) of glucagon. The fractional saturation of RI and RII was always similar, being 20% in nonstimulated cells. 50% saturation occurred when free cAMP was 0.46 pmol/10(5) cells. A Scatchard plot of the data for the endogenous cAMP binding suggested that cAMP interacted with RI and RII in a slightly positively cooperative manner. About 5% of the intracellularly bound cAMP was sedimentable at 10,000 X g. The apparent affinity of these particulate-associated binding sites was similar to that of soluble RI and RII. Under the conditions used no evidence was obtained for cAMP binding to other proteins than RI and RII. PMID- 2982858 TI - A monoclonal antibody which inhibits epidermal growth factor binding has opposite effects on the biological action of epidermal growth factor in different cells. AB - An epidermal growth factor (EGF) receptor-interactive monoclonal antibody (151 IgG) that inhibits EGF binding to PC12 rat pheochromocytoma cells and to various other cell types has been produced. The hybridoma clone was obtained by fusing Sp2/O-Ag14 myeloma cells with splenocytes from Balb/C mice which had been immunized with n-octyl glucoside-solubilized protein from isolated PC12 cell plasma membranes. The antibody is an IgG which binds to protein A. 151-IgG did not bind EGF. At 0.5 degrees C 151-IgG was directly competitive for EGF binding to PC12 cells. It also inhibited EGF binding to bovine corneal endothelial cells, rabbit corneal fibroblasts, human foreskin fibroblasts, and normal rat kidney cells, and it slightly enchanced EGF binding to SW 3T3 cells. PC12 cells have the same number of binding sites for 151-IgG as for EGF (approximately 27,000 sites/cell). 151-IgG inhibited the photoactivatable cross-linking of EGF to a protein of Mr 170,000 in PC12 cells. 151-IgG inhibited the EGF-stimulated incorporation of [3H]thymidine into quiescent bovine corneal endothelial cells, rabbit corneal endothelial cells, epithelial normal rat kidney cells, and SW 3T3 cells while it enhanced the EGF-stimulated [3H]thymidine incorporation into quiescent human foreskin fibroblasts. 151-IgG by itself possessed intrinsic EGF like activity for human fibroblasts but not for the other cells tested. This suggests that there is a difference in EGF receptors and/or processing in these normal cell types. PMID- 2982861 TI - The effect of ligand heterogeneity on the Scatchard plot. Particular relevance to lipoprotein binding analysis. AB - Computer simulations of equilibrium binding studies of a mixture of two labeled ligands binding competitively to a single class of identical and independent sites (receptors) were performed to investigate how ligand heterogeneity affects the observed data in such studies. The simulated data are presented in Scatchard plots. Ligand heterogeneity was generally found to be indistinguishable from the case of a homogeneous ligand when usual experimental conditions applied (that is, Scatchard plots of the data were straight lines). Some factors that increased the probability of recognizing heterogeneity in the system were identified, however. These are 1) a large difference between the dissociation constants of the two ligands, 2) a high concentration of receptors relative to the dissociation constant of the higher-affinity ligand, 3) a high concentration of the lower affinity ligand relative to that of the higher-affinity ligand, 4) a high specific activity of the lower-affinity ligand relative to that of the higher affinity ligand, and 5) lack of experimental error. When ligand heterogeneity (under certain conditions) did cause curvilinearity in the Scatchard plot, the curve formed was always concave-downwards. Thus, ligand heterogeneity may occasionally mimic positive cooperativity, but never mimics negative cooperativity or multiple classes of binding sites. Implications of these findings for equilibrium binding studies involving lipoproteins (which are generally isolated as heterogeneous mixtures of particles) are discussed in detail. These findings are also relevant to equilibrium binding studies using ligands which are mixtures of stereoisomers or which contain chemical or radiochemical impurities. PMID- 2982860 TI - The regulatory subunit monomer of cAMP-dependent protein kinase retains the salient kinetic properties of the native dimeric subunit. AB - Monomeric regulatory subunit (R) fragments of type II cAMP-dependent protein kinase were compared with the parent dimeric R. The monomeric fragments were generated by either endogenous proteolysis of rabbit muscle R or by trypsin treatment of bovine heart R in the holoenzyme form. During isolation of pure R from rabbit muscle, carboxyl-terminal fragments of Mr = 42,000 (42 K) and Mr = 37,000 by denaturing gels are generated by endogenous proteolysis. Although the autophosphorylation site is retained, the 42 K is not dimeric (as is its native 56 K precursor) but, in contrast to the monomeric 37 K product, actively reassociates with purified catalytic subunit (C). Several lines of evidence indicate a type II R origin of the 42 K. N-terminal sequence analysis of the 42 K shows some homology with known bovine RI, RII, and cGMP-dependent protein kinase sequences. Both cyclic nucleotide-binding sites (two/42 K or 37 K) and the site selectivity of cAMP analogs are retained in the monomeric fragments. When purified bovine heart holoenzyme, which contains a dimeric Mr = 56,000 R (denaturing gel analysis) and two C subunits, is treated with trypsin followed by separation procedures, the product is a fully recovered active enzyme with an unaltered ratio of cAMP binding to catalytic activity. From Mr considerations, the product is a dimer containing one intact C and a proteolyzed R of Mr = 48,000 on denaturing gels. This dimeric enzyme is not significantly different from the parent tetramer in cAMP concentration dependence (Hill constant = 1.63), [3H]cAMP dissociation behavior (both intrasubunit cAMP-binding sites are present), stimulation of [3H]cIMP binding by site-selective cAMP analogs, and synergism between two analogs in kinase activation. The data indicate that 1) proteolytic cleavage of the native R dimer can cause monomerization without appreciably affecting the inhibition of C and 2) essentially all of the cAMP binding cooperativity is an intrasubunit interaction. PMID- 2982862 TI - Regions of the terminal repetitions of the herpes simplex virus type 1 genome. Relationship to immunoglobulin switch-like DNA sequences. AB - Several recombinant clones isolated from a mouse genomic library were previously shown to hybridize with a SmaI fragment located in the terminal repetition of the S component of herpes simplex virus DNA. We report here the nucleotide sequence of the related regions in two mouse clones, TGL19 and TGL35, as well as that of the SmaI fragment of HSV-1. The mouse DNA clones have a core of repetitive sequences 80% homologous to a tandem repeat (reiteration II) in the viral fragment. The regions of homology are in turn related to immunoglobulin class switch sequences, due mostly to the presence of the pentamer TGGG(G), involved in class-switch recombination. These results suggest that the HSV genome has recombination sequences identical to those of the host cell and provide a possible explanation for the high frequency of recombination events observed in this region of the viral genome. PMID- 2982863 TI - Phosphorolytic cleavage of diadenosine 5',5'''-P1,P4-tetraphosphate. Properties of homogeneous diadenosine 5',5'''-P1,P4-tetraphosphate alpha, beta-phosphorylase from Saccharomyces cerevisiae. AB - Novel enzymatic activity which splits diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) phosphorolytically has been found in extracts from Saccharomyces cerevisiae. One of the two alpha,beta-anhydride bonds between Ap4A phosphate residues undergoes phosphorolysis, and ATP (pppA) plus ADP (ppA) are the products of the reaction according to the equation: AppppA + P*i----pppA + p*pA The reaction is dependent on the presence of divalent metal ions; Mn2+ or Mg2+ sustain the greatest rates of reaction. Among analogues of the Ap4A substrate, Ap5A and Gp4G, but not p4A and Ap3A, are substrates, and corresponding products are p4A plus ADP, and GTP plus GDP, the phosphate being incorporated into the nucleoside 5'-diphosphates. In the reactions, phosphate can be substituted with arsenate. Arsenolysis of Ap4A, Ap5A, or Gp4G leads to ATP plus AMP, p4A plus AMP, and GTP plus GMP, respectively. The name diadenosine tetraphosphate alpha,beta phosphorylase (ADP-forming) is proposed for the new enzyme. The phosphorylase has been purified to apparent homogeneity and behaves as a single polypeptide chain of Mr = 40,000. Optimum activity of the enzyme is at pH 8.0 and the sulfhydryl groups are essential for catalysis. At saturating Ap4A, the rate constant for the reaction is 36 s-1 and the Km value for Ap4A is 60 microM (37 degrees C, 50 mM Hepes/KOH (pH 8.2), 500 microM MnCl2, 10 mM K2HPO4, 1 mM 2-mercaptoethanol, and 2% glycerol). The Km values for phosphate and arsenate are 1 and 3 mM, respectively. PMID- 2982864 TI - Leukotriene formation by a purified reticulocyte lipoxygenase enzyme. Conversion of arachidonic acid and 15-hydroperoxyeicosatetraenoic acid to 14, 15-leukotriene A4. AB - The purified lipoxygenase of rabbit reticulocytes converts arachidonic acid at 0 degrees C to 15-hydroperoxyeicosatetraenoic acid (15-HPETE) and to 12 hydroperoxyeicosatetraenoic acid (12-HPETE) via reactions which involve hydrogen abstraction at C-13 and C-10, respectively. At 37 degrees C the enzyme converts arachidonic acid to additional products which were identified as 13-hydroxy-14,15 epoxy-5,8,11-eicosatrienoic acid, 8,15-dihydroperoxy-5,9,11,13- and 5,15 dihydroperoxy-6, 6,8,11,13-eicosatetraenoic acids (8,15-diHPETE and 5,15-HPETE, respectively) and diastereoisomers of 8,15-dihydroxy-5,9,11,13-eicosatetraenoic acid (8,15-diHPETEs). The 8,15- and 5,15-diHPETEs were formed by double lipoxygenation since each incorporated 2 molecules of 18O2 and since their synthesis from 15-HPETE was blocked under anaerobic conditions. The 8,15-diHETEs each incorporated 18O from 18O2 at C-15 and were found to arise from nonenzymatic hydrolysis of an epoxytriene which was identified as 14,15-leukotriene A4 by trapping in acidic methanol. This compound was a major product of 15-HPETE in anaerobic incubations. The conversion of 15-HPETE to 14,15-leukotriene A4 was inhibited by the lipoxygenase inhibitors nordihydroguairetic acid and 5,8,11,14 eicosatetraynoic acid. The 14,15-leukotriene A4 synthase and 15-lipoxygenase activities were inhibited by 5,8,11,14-eicosatetraynoic acid in a similar time dependent manner. The results support a mechanism whereby 14,15-leukotriene A4 is synthesized from 15-HPETE by a further enzymatic step carried out by the reticulocyte 15-lipoxygenase via hydrogen abstraction at C-10 and a redox cycle of the non-heme iron atom of the enzyme. PMID- 2982865 TI - Multiple affinity states of opiate receptor in neuroblastoma x glioma NG108-15 hybrid cells. Opiate agonist association rate is a function of receptor occupancy. AB - The existence of multiple affinity states for the opiate receptor in neuroblastoma x glioma NG108-15 hybrid cells has been demonstrated by competition binding studies with tritiated diprenorphine and [D-Ala2, D-Leu5]enkephalin (DADLE). In the presence of 10 mM Mg2+, all receptors exist in a high affinity state with Kd = 1.88 +/- 0.16 nM. Addition of 10 microM guanyl-5'-yl imidodiphosphate (Gpp(NH)p) decreased the affinity of DADLE to Kd = 8.08 +/- 0.93 nM. However, in the presence of 100 mM Na+, which is required for opiate inhibition of adenylate cyclase activity, analysis of competition binding data revealed three sites: the first, consisting of 17.5% of total receptor population has a Kd = 0.38 +/- 0.18 nM; the second, 50.6% of the population, has a Kd = 6.8 +/- 2.2 nM; and the third, 31.9% of the population, has a Kd of 410 +/- 110 nM. Thus, in the presence of sodium, a high affinity complex between receptor (R), GTP binding component (Ni), and ligand (L) was formed which was different from that formed in the absence of sodium. These multiple affinity states of receptor in the hybrid cells are agonist-specific, and the percentage of total opiate receptor in high affinity state is relatively constant in various concentrations of Na+. Multiple affinity states of opiate receptor can be demonstrated further by Scatchard analysis of saturation binding studies with [3H]DADLE. In the presence of Mg2+, or Gpp(NH)p, analysis of [3H]DADLE binding demonstrates that opiate receptor can exist in a single affinity state, with apparent Kd values of [3H]DADLE in 10 mM Mg2+ = 1.75 +/- 0.28 nM and in 10 microM Gpp(NH)p = 0.85 +/- 0.12 nM. There is a reduction of Bmax value from 0.19 +/- 0.02 nM in the presence of Mg2+ to 0.14 +/- 0.03 nM in the presence of Gpp(NH)p. In the presence of 100 mM Na+, Scatchard analysis of saturation binding of [3H]DADLE reveals nonlinear plots; two-site analysis of the curves yields Kd = 0.43 +/- 0.09 and 7.9 +/- 3.2 nM. These Kd values are analogous to that obtained with competition binding studies. Again, this conversion of single site binding Scatchard plots to multiple sites binding plots in the presence of Na+ is restricted to 3H-agonist binding only.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982866 TI - Identification of apolipoproteins involved in the interaction of human high density lipoprotein3 with receptors on cultured cells. AB - Human high density lipoprotein (HDL), devoid of apolipoproteins E or B, binds with high affinity and specificity to cultured cells derived from several tissues. In order to investigate the ligand specificity of the putative receptor, we have performed competitive inhibition studies to identify the components of high density lipoprotein that bind to cell surfaces of rat adrenal cortical cells and human skin fibroblasts. Radiolabeled HDL3 was displaced with unlabeled apolipoprotein-dimyristoylphosphatidylcholine recombinant particles containing AI, AII, CIII-1, and E apolipoproteins, but not by dimyristoylphosphatidylcholine complexed to albumin or by low density lipoprotein. Because exchange may readily occur between apolipoproteins in HDL and in recombinants this observation may not be truly representative of ligand competition. Further experiments using Fab fragments prepared from pure IgG to each apolipoprotein showed that binding of radioiodinated HDL to cells was suppressed following preincubation of HDL with Fab fragments raised against apolipoproteins AI or AII but not against apolipoproteins E or CIII-1 or albumin. In additional studies with apolipoprotein recombinants specific saturable binding was demonstrated between apo-AI or -AII recombinants and adrenocortical cells whereas binding of apo-CIII-2 was characterized by a large nonsaturable component which almost equaled the specific binding. The data, therefore, provide evidence for the involvement of the two major apolipoproteins (AI and AII) in HDL recognition by cellular receptors. PMID- 2982867 TI - The role of enzyme sequestration in the regulation of the adenylate cyclase of Dictyostelium discoideum. AB - Although the adenylate cyclase of Dictyostelium discoideum cannot be activated by its cAMP agonist in vitro, its in vivo activation can be demonstrated by rapidly breaking and assaying the cells, over 10-fold higher activity being observed for stimulated cells than for basal cells. We report here that when basal cells are broken in the presence of labeled ATP and then rapidly assayed, they display 8 fold more adenylate cyclase activity than cells broken in the presence of unlabeled ATP. This suggests that a significant amount of the enzyme in extracts of basal cells is sequestered within vesicles that can be loaded with substrate at the time of cell lysis, but then rapidly seal. In contrast to the results obtained with basal cells, when cells activated in vivo are broken in the presence of labeled ATP, there is less than 2-fold increase in adenylate cyclase activity. Thus, a much smaller percentage of the observed adenylate cyclase activity of stimulated cells appears to be due to sequestered enzyme than of basal cells. Two models are discussed that account for these observations. One model envisions that roughly equal populations of sequestered and nonsequestered enzyme are produced upon breakage of both basal and activated cells, but that sequestered enzyme in basal extracts becomes uniquely activated in vitro. The other model proposes that the differences in observed activity are due directly to differences in sequestration. According to this latter model, nearly all of the -fold activation previously observed for the D. discoideum adenylate cyclase can be accounted for by a change in sequestration of the enzyme rather than by an intrinsic alteration in the enzyme per se. It therefore suggests a novel mode of regulation whereby an enzyme may be packaged within vesicles and its activity controlled by modulating the permeability of the vesicles to its substrate or effectors. PMID- 2982868 TI - Progesterone-inhibited phosphorylation of an unique Mr 48,000 protein in the plasma membrane of Xenopus laevis oocytes. AB - The meiotic maturation of Xenopus laevis oocytes is induced in vitro by progesterone which interacts at the cell surface level. A cell-free membrane preparation (P-10,000) incorporated 32P from [gamma-32P]ATP, mostly into two proteins, Mr approximately 56,000 and approximately 48,000 (as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Progesterone, added in vitro, specifically inhibited the phosphorylation of the Mr approximately 48,000 protein (named p48). Half-maximal inhibition of p48 phosphorylation occurred with progesterone approximately 8 microM, in good correlation with hormone concentration inducing oocyte maturation. The effect was not due to stimulation of protein phosphatase activity. The potent maturation inducers testosterone and deoxycorticosterone also inhibited p48 phosphorylation, whereas biologically inactive steroids or cholesterol did not. p48 phosphorylation was not affected by cAMP, cGMP, polyamines, calmodulin, and phospholipids + diolein. EGTA had a stimulatory effect which was reversed by added Ca2+. The inhibitory effects of progesterone and Ca2+ were additive, suggesting two distinct sites of action. Phospho-p48 was not detected in yolk platelets, microsomes, and cytosol of oocytes. Contrary to p48 itself, the p48 kinase activity was loosely associated with P-10,000. Progesterone inhibited p48 phosphorylation produced by either cytosol or exogenous pure catalytic subunit of cAMP-dependent protein kinase. Conversely, phosphorylation of casein and histones by protein kinase activity present in P-10,000 was not modified by progesterone. It is then suggested that progesterone regulates p48 phosphorylation by affecting the protein substrate in the membrane, rather than by inhibiting the protein kinase enzyme itself. The data demonstrate a direct effect (not mediated by change of protein synthesis) of steroids on p48 phosphorylation in the plasma membrane, and they suggest that this protein could be implicated in the initial action of progesterone on oocyte maturation. PMID- 2982869 TI - Proton translocation by a native and subunit III-depleted cytochrome c oxidase reconstituted into phospholipid vesicles. Use of fluorescein phosphatidylethanolamine as an intravesicular pH indicator. AB - The existence of a proton pump associated with bovine cytochrome c oxidase (EC 1.9.3.1) has over the last few years been a matter of considerable dispute. In an attempt to resolve some of the problems with the measuring system we have synthesized fluorescein-phosphatidylethanolamine which when reconstituted with cytochrome c oxidase into phospholipid vesicles provided a reliable indicator of the intravesicular pH. It was observed that cytochrome c oxidase catalyzed the abstraction of almost 2 protons from the intravesicular medium/molecule of ferrocytochrome c oxidized. In parallel experiments whereby the extravesicular pH was measured with an electrode it was found that the enzyme appeared to be responsible for the appearance of almost 1.0 proton/molecule of ferrocytochrome c oxidized. Taken together these data unequivocally demonstrate that cytochrome c oxidase behaves as a proton pump. Furthermore, the other proton which was abstracted is believed to be used for the process of the reduction of oxygen. Similar experiments were performed with a cytochrome c oxidase preparation which was devoid of subunit III. Under these circumstances the enzyme appeared to be unable to translocate protons across the vesicular membrane but was competent to abstract protons from the intravesicular medium for the reduction of oxygen. PMID- 2982870 TI - Enhancement by Ca2+ or Mg2+ of catalytic activity of the superoxide-producing NADPH oxidase in membrane fractions of human neutrophils and monocytes. AB - To examine the role of divalent cations in the generation of superoxide anion (O2 ) by the NADPH oxidase system of phagocytic cells, membrane-rich fractions were prepared from human neutrophils and monocytes. O2- generation by the fractions in sucrose was enhanced by addition of Ca2+ or Mg2+. EDTA inhibited most of the O2- generation; Ca2+ or Mg2+ reversed the inhibition. Zn2+, Mn2+, or Cu2+ completely inhibited O2- production. Neutrophil membrane fraction solubilized with Triton X 100, then passed through a chelating column, lost 80% of its oxidase activity; the loss could be reversed by addition of Ca2+ or Mg2+. Addition of 0.3 mM Ca2+ or Mg2+ protected against thermal instability of the enzyme. Kinetic analysis of the neutrophil oxidase activity as a function of NADPH and Ca2+ or Mg2+ concentrations showed that cation did not interact with NADPH in solution or affect the binding of NADPH to the oxidase; rather, cation bound directly to the oxidase, or to some associated regulatory component, to activate the enzyme. For the neutrophil oxidase, the Km for NADPH was 51 +/- 6 (S.D.) microM. Hyperbolic saturation was observed with Ca2+ and Mg2+, and the Kd values were 1.9 +/- 0.3 and 2.9 +/- 0.3 microM, respectively, suggesting that the oxidase, or some associated component, has a relatively high-affinity binding site for Ca2+ and Mg2+. PMID- 2982872 TI - Compartmentalization of the proteins encoded by IS50R. AB - IS50R is a transposable genetic element that serves as the right inverted repeat of the transposon Tn5. Earlier work has shown that IS50R encodes at least two proteins (called P1 and P2) involved in transposition. In this paper, we describe the localization properties of the proteins encoded on this repeat. Strains were constructed that overproduced either these two proteins or hybrids between beta galactosidase and the IS50R proteins. An antiserum was raised against the hybrid proteins, and this was used to study the localization of P1 and P2. Based on studies in maxicells as well as in growing cells, we show that P1 and P2 are localized differently in the cell. P2 is a cytoplasmic protein, while P1 largely fractionates with the membrane. PMID- 2982871 TI - Adrenocorticotropic stimulation and insulin inhibition of adipocyte phospholipid methylation. AB - Treatment of isolated rat adipocytes with adrenocorticotropin (ACTH) caused a 1.5 fold increase in phospholipid methyltransferase activity within 5 min. This effect of ACTH was concentration-dependent with maximal activation at 2 milliunits/ml ACTH, and was reproduced by dibutyryl cyclic AMP. ACTH (2 milliunits/ml) caused an increase in the Vmax value of phospholipid methyltransferase without changing the Km for S-adenosyl-L-methionine. Insulin caused a concentration-dependent inhibition of both control and ACTH-stimulated phospholipid methyltransferase. Half-maximal inhibition by insulin was demonstrated with 5 microunits/ml insulin in control cells and with 25 microunits/ml insulin in ACTH-stimulated cells. The rapid and sensitive activation of adipocyte phospholipid methyltransferase by ACTH and inhibition by insulin are consistent with a role for this pathway in the hormonal response of the adipocyte. PMID- 2982873 TI - Isolation and analysis of an Abelson murine leukemia virus-encoded tyrosine specific kinase produced in Escherichia coli. AB - A segment of the coding sequence of the Abelson murine leukemia virus transforming gene (v-abl) has been inserted into a plasmid vector that allows its efficient and regulated expression in Escherichia coli. The product of the v-abl derived coding sequence, designated p60v-abl, accumulated to a level of approximately 10% of total E. coli protein. A procedure is described for the isolation of p60v-abl from E. coli that yields about 50 micrograms of p60v-abl/g wet weight of E. coli. p60v-abl was capable of autophosphorylation and phosphorylating certain E. coli proteins specifically at tyrosine residues. The E. coli-expressed p60v-abl specifically phosphorylated tyrosine residues on casein and angiotensin II. The Km and Vmax values for ATP, casein, and angiotensin II in the p60v-abl kinase reaction have been determined and compared to values reported for other tyrosine-specific kinases. The expression system and isolation procedure described here permit the preparation of functional p60v-abl in quantities sufficient for detailed physical and biochemical characterization and examination of its biological action(s). PMID- 2982874 TI - A linear double-stranded RNA in Trichomonas vaginalis. AB - A "double-stranded" RNA was identified in the anaerobic, parasitic protozoan Trichomonas vaginalis. Electron microscopic evidence indicated linear double stranded structure 1.5 micron in length, with no apparent hairpins or loops. Boiling in 30% dimethyl sulfoxide denatured it into single strands of 1.5 micron and shorter fragments. It consists of 23.4% G, 23.4% C, 23.0% A, and 30.3% U and melts at a transition temperature of 81.7 degrees C in 75 mM NaCl and 7.5 mM sodium citrate, pH 7.0, with 7-15% hyperchromicity. The 32P-labeled double stranded RNA hybridized specifically with T. vaginalis DNA fragments in a single DNA band from EcoRI digest and two DNA bands from HindIII digest. Of the 33 different strains or isolates of T. vaginalis examined, all contained this double stranded RNA. However, the only two metronidazole-resistant T. vaginalis strains examined thus far (IR78 and 85) contained no detectable double-stranded RNA, although the corresponding DNA sequence was present. DNA fragments of Escherichia coli and Giardia lamblia did not hybridize with the double-stranded RNA. But DNA fragments of a metronidazole-sensitive Tritrichomonas foetus hybridized specifically with the double-stranded RNA, even though this organism does not contain the double-stranded RNA itself. PMID- 2982875 TI - Spectroscopic and functional properties of subunit III-depleted cytochrome oxidase. AB - Beef heart cytochrome c oxidase has been depleted of subunit III by treatment with chymotrypsin. The removal of subunit III has been evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel fluorography of preparations of the oxidase labeled with [14C]dicyclohexylcarbodiimide prior to proteolysis. Removal of subunit III resulted in a perturbation of the visible spectrum of reduced cytochrome oxidase. Subunit III-depleted oxidase is spectroscopically very similar to the oxidase from Paracoccus denitrificans. When reconstituted into liposomes, the depleted enzyme still pumped protons in response to a pulse of reduced cytochrome c. The H+/e- stoichiometry averaged 0.5. Redox-linked proton translocation could be observed only when respiratory control ratios were higher than 3 and the reductant pulse was of a magnitude that allowed for no more than 5 turnovers of the oxidase. PMID- 2982876 TI - Properties of the globular domain of type IV collagen and its relationship to the Goodpasture antigen. AB - The globular domain of type IV collagen from bovine glomerular basement membrane was solubilized by collagenase digestion. Components of this domain include several monomer-size and structurally related dimer-size polypeptides. The monomer-size polypeptides were resolved into three fractions (M1, M2, and M3) with slightly different mobilities upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (nonreduced Mr = 24,500-28,300). Chemical and immunochemical studies indicate that each is a distinct component. M2 is reactive with antibodies from patients with Goodpasture syndrome. The molecular weight by sedimentation equilibrium was 32,000 for M2 and 28,000 for M1. The dimers were characterized as two classes, D1 and D2. D1 consists of two sets of nonreactive components (D1a-d and D1a,c) whereas D2 contains one set of four components (D2a d), each of which is reactive with Goodpasture sera. Chemical and immunochemical studies indicate that a monomer-dimer relationship exists between M1 and D1 and between M2 and D2. The origin of M3 remains undetermined. Rabbit antibodies to type IV collagen alpha chains react with M1 and M2, and antibodies to M1 and M2 react with type IV collagen alpha chains, which provides additional evidence for the localization of the Goodpasture antigen to one of the chains of type IV collagen. PMID- 2982877 TI - Effect of butyrate on the expression of microinjected or transfected genes. AB - We have studied the effect of sodium n-butyrate on the expression of specific genes. For this purpose, tk-ts13 cells (a thymidine kinase-deficient mutant originating from Syrian hamster cells) were microinjected or transfected with pC2, a plasmid containing the entire SV40 genome and the herpes simplex virus thymidine kinase gene (HSV-TK), cloned in pBR322. As a measure of the expression of these two genes, one of which is spliced (SV40) and the other one (HSV-TK) which is not, we have taken the protein levels (amount of T antigen for SV40 and incorporation of [3H]thymidine for HSV-TK) and the levels of RNA (by dot blot hybridization). The expression of the microinjected genes was inhibited when tk ts13 cells were exposed to butyrate, actinomycin D, cycloheximide, and mitomycin C, but not when the cells were treated with insulin or dexamethasone. Further studies showed that a decrease in the percentage of T-positive cells occurs at lower concentrations of butyrate than a decrease in the levels of specific mRNA. In tk-ts13 cells transfected with pC2 and treated with butyrate at a concentration of 3 mM, SV40 mRNA levels are not decreased but the percentage of T positive cells is decreased 50%. At 5 mM, the amount of T antigen/cell is decreased a further 40%. These results indicate that butyrate may have at least two sites of action, one at the level of mRNA amount and a second at the level of protein amount. In addition, our studies show that the use of microinjected or transfected genes offers certain unique possibilities for studies on the effects of environmental manipulations on gene expression. PMID- 2982879 TI - Malignant fibrous histiocytoma of bone. AB - Malignant fibrous histiocytoma is a rare primary bone tumor, and there have been conflicting reports on its grades of malignancy. We are describing the cases of eight patients who were seen between 1977 and 1982. Four had pulmonary metastases and five, involvement of the lymph nodes. Five patients had a high level of serum alkaline phosphatase. None of the patients had a pathological fracture, an associated bone infarct, or Paget's disease. Seven of the eight patients died within one year after diagnosis. PMID- 2982878 TI - Isoproterenol-induced phosphorylation of a 15-kilodalton sarcolemmal protein in intact myocardium. AB - The effect of beta-adrenergic stimulation on sarcolemmal protein phosphorylation was examined in intact ventricular myocardium. Isolated guinea pig ventricles were perfused via the coronary arteries with 32Pi after which membrane vesicles enriched 3-5-fold in sarcolemma were isolated by differential centrifugation followed by sucrose gradient centrifugation. Perfusion of hearts with isoproterenol stimulated 32P incorporation into a protein of apparent molecular weight of 15,000, which copurified with sarcolemmal vesicles. The increase in 32P incorporation was rapid in onset and elevated 2.5-3.0-fold after 30-45 s exposure of hearts to 100 nM isoproterenol. A positive correlation was found between stimulation of phosphorylation of the 15-kDa protein and the increase in the maximal rate of developed tension in intact ventricles after administration of isoproterenol. Phosphorylated phospholamban (most likely present as a contaminant) was also identified in the same sarcolemmal preparations. However, phospholamban and the 15-kDa sarcolemmal substrate were different proteins. Boiling of the membrane samples in sodium dodecyl sulfate prior to electrophoresis dissociated the high Mr form of phospholamban into the form of lower Mr but did not alter the mobility of the 15-kDa protein in sodium dodecyl sulfate-polyacrylamide gels. The 15-kDa protein did not undergo the electrophoretic mobility shift that is characteristic of phospholamban after cAMP dependent phosphorylation nor did it cross-react with a highly specific phospholamban antibody. In vitro phosphorylation experiments conducted with the unmasking agent Triton X-100 suggested that the 15-kDa protein was localized to the cytoplasmic surfaces of sarcolemmal vesicles. These results demonstrate phosphorylation of a sarcolemmal protein, distinct from phospholamban, in response to beta-adrenergic stimulation of the heart. Phosphorylation of the sarcolemmal 15-kDa protein may play a role in mediating the effects of beta adrenergic agonists on cardiac contractile force. PMID- 2982880 TI - Distribution of fibronectin and laminin in human liver tumors. AB - Two extracellular matrix and basement-membrane components, fibronectin (Fn) and laminin, were studied by the indirect immunoperoxidase technique in ten primary human liver cancers. Similar distributions of both Fn and laminin were detected in the well differentiated hepatocellular carcinomas with trabecular and tubular pattern. Two moderately differentiated hepatomas contained Fn only. Neither Fn nor laminin were present, however, in the parenchyma of one poorly differentiated hepatoma. In three cases of cholangiocarcinoma, laminin surrounded the tumorous ducts, while Fn appeared mainly in the reactive connective tissue stroma. The present findings indicate that bile-duct cancers synthesize laminin, and not Fn while differentiated hepatocellular carcinomas produce both Fn and laminin in vivo. The presence of Fn even in moderately differentiated types of liver cancer is in contrast to the findings for carcinomas developing from other organs and it may serve as a marker for primary hepatocellular carcinomas in the differential diagnosis. PMID- 2982881 TI - The early appearance of fibronectin in the course of metastatic tumor growth in lymph nodes. AB - In order to study structural changes in the lymph node architecture in the course of metastatic involvement by carcinoma, 70 axillary lymph nodes of 45 female patients affected by breast carcinoma were examined. The following growth pattern of the metastasis was observed: (1) Involvement of vasa afferentia and (marginal) sinuses; (2) lysis of sinus wall and infiltration, causing replacement and lysis of reticulin fibres; (3) appearance of fibrillary fibronectin (FN) as the first noticeable sign of tumor stroma; (4) immigration of fibroblasts along these fibres, appearance of argyrophilic fibres codistributed with fibrillary FN; (5) formation of the complete tumor stroma, neovessels included. The appearance of FN is thus the first step of stroma formation and is interpreted as an "in situ" precipitation of soluble plasma FN. PMID- 2982882 TI - Small-cell lung cancer and immunochemotherapy with Propionibacterium granulosum KP 45. AB - Seventy-nine patients with small-cell lung cancer were treated with vincristin, methotrexate, and cyclophosphamide in inductive therapy and with methotrexate, cyclophosphamide, and procarbazine in maintenance therapy. Patients were divided at random into two groups: one group received chemotherapy alone and the second group was additionally subjected to systemic immunotherapy with Propionibacterium granulosum strain KP-45. In general, differences in the frequency of therapy response and in duration of remission could not be stated between the two groups of patients, but patients responding to chemotherapy showed a significantly longer remission time and lower complication rates. This benificial effect of chemoimmunotherapy is not related to a direct antitumor activity of the immunomodifier used, but to the lowered risk of myelosuppression and infections. Immunomodulation in combination with chemo- and/or radiotherapy can be recommended for the treatment of small-cell lung cancer. PMID- 2982883 TI - Tropomyosin isoforms in chicken embryo fibroblasts: purification, characterization, and changes in Rous sarcoma virus-transformed cells. AB - Seven polypeptides (a, b, c, 1, 2, 3a, and 3b) have been previously identified as tropomyosin isoforms in chicken embryo fibroblasts (CEF) (Lin, J. J.-C., Matsumura, F., and Yamashiro-Matsumura, S., 1984, J. Cell. Biol., 98:116-127). Spots a and c had identical mobility on two-dimensional gels with the slow migrating and fast-migrating components, respectively, of chicken gizzard tropomyosin. However, the remaining isoforms of CEF tropomyosin were distinct from chicken skeletal and cardiac tropomyosins on two-dimensional gels. The mixture of CEF tropomyosin has been isolated by the combination of Triton/glycerol extraction of monolayer cells, heat treatment, and ammonium sulfate fractionation. The yield of tropomyosin was estimated to be 1.4% of total CEF proteins. The identical set of tropomyosin isoforms could be found in the antitropomyosin immunoprecipitates after the cell-free translation products of total poly(A)+ RNAs isolated from CEF cells. This suggested that at least seven mRNAs coding for these tropomyosin isoforms existed in the cell. Purified tropomyosins (particularly 1, 2, and 3) showed different actin-binding abilities in the presence of 100 mM KCl and no divalent cation. Under this condition, the binding of tropomyosin 3 (3a + 3b) to actin filaments was significantly weaker than that of tropomyosin 1 or 2. CEF tropomyosin 1, and probably 3, could be cross-linked to form homodimers by treatment with 5,5'-dithiobis-(2 nitrobenzoate), whereas tropomyosin a and c formed a heterodimer. These dimer species may reflect the in vivo assembly of tropomyosin isoforms, since dimer formation occurred not only with purified tropomyosin but also with microfilament associated tropomyosin. The expression of these tropomyosin isoforms in Rous sarcoma virus-transformed CEF cells has also been investigated. In agreement with the previous report by Hendricks and Weintraub (Proc. Natl. Acad. Sci. USA., 78:5633-5637), we found that major tropomyosin 1 was greatly reduced in transformed cells. We have also found that the relative amounts of tropomyosin 3a and 3b were increased in both the total cell lysate and the microfilament fraction of transformed cells. Because of the different actin-binding properties observed for CEF tropomyosins, changes in the expression of these isoforms may, in part, be responsible for the reduction of actin cables and the alteration of cell shape found in transformed cells. PMID- 2982884 TI - cAMP induces a rapid and reversible modification of the chemotactic receptor in Dictyostelium discoideum. AB - Stimulation, within 1 min after cAMP stimulation, of aggregation-competent Dictyostelium discoideum amebae was found to cause a rapid (within 1 min) modification of the cell's surface cAMP receptor. The modified receptor migrated on SDS PAGE as a 47,000-mol-wt protein, as opposed to a 45,000-mol-wt protein labeled on unstimulated cells. The length of time this modified receptor could be detected depended upon the strength of the cAMP stimulus: 3-4 min after treatment with 10(-7) M cAMP, cells no longer possessed the 47,000-mol-wt form of the cAMP receptor. Instead, the 45,000-mol-wt form was present. Stimulation of cells with 10(-5) M cAMP, however, resulted in the persistent (over 15 min) expression of the modified receptor. The time course, concentration dependence, and specificity of stimulus for this cAMP-induced shift in the cAMP receptor were found to parallel the cAMP-stimulated phosphorylation of a 47,000-mol-wt protein. In addition, both phenomena were shown to occur in the absence of endogenous cAMP synthesis. The possibility that the cAMP receptor is phosphorylated in response to cAMP stimulation, and the role of this event in cell desensitization, are discussed. PMID- 2982885 TI - Intracellular movement of cell surface receptors after endocytosis: resialylation of asialo-transferrin receptor in human erythroleukemia cells. AB - The intracellular movement of cell surface transferrin receptor (TfR) after internalization was studied in K562 cultured human erythroleukemia cells. The sialic acid residues of the TfR glycoprotein were used to monitor transport to the Golgi complex, the site of sialyltransferases. Surface-labeled cells were treated with neuraminidase, and readdition of sialic acid residues, monitored by isoelectric focusing of immunoprecipitated TfR, was used to assess the movement of receptor to sialyltransferase-containing compartments. Asialo-TfR was resialylated by the cells with a half-time of 2-3 h. Resialylation occurred in an intracellular organelle, since it was inhibited by treatments that allow internalization of surface components but block transfer out of the endosomal compartment. Moreover, roughly half of the resialylated molecules were cleaved when cells were retreated with neuraminidase after culturing, indicating that this fraction of the molecules had returned to the cell surface. These results suggest that TfR is transported from the cell surface to the Golgi complex, the intracellular site of sialyltransferases, and then returns to the cell surface. This pathway, which has not been previously described for a cell surface receptor, may be different from the route followed by TfR in iron uptake, since reported rates of transferrin uptake and release are significantly more rapid than the resialylation of asialo-TfR. PMID- 2982887 TI - Synergistic effects of cyclic AMP and nerve growth factor on neurite outgrowth and microtubule stability of PC12 cells. AB - The outgrowth of neurites from rat PC12 cells stimulated by combined treatment of nerve growth factor (NGF) with cAMP is significantly more rapid and extensive than the outgrowth induced by either factor alone. We have compared the responses of PC12 cells under three different growth conditions, NGF alone, cAMP alone, and combined treatment, with respect to surface morphology, rapidity of neurite outgrowth, and stability of neurite microtubules, to understand the synergistic action of NGF and cAMP on PC12. Surface events at early times in these growth conditions varied, suggesting divergent pathways of action of NGF and cAMP. This suggestion is strongly supported by the finding that cells exposed to saturating levels of dibutyryl cAMP without substantial neurite outgrowth initiated neurites within 5 min of NGF. This response has been adopted as a convenient assay for NGF. Neurites that regenerated in the three growth conditions showed marked differences in stability to treatments that depolymerize microtubules. The results indicate that microtubules in cells treated with both NGF and cAMP are significantly more stable than in either growth factor alone. We suggest that a shift of the assembly equilibrium favoring tubulin assembly is a necessary prerequisite for the initiation of neurites by PC12. PMID- 2982886 TI - Phosphorylation-dependent subcellular translocation of a Ca2+/calmodulin dependent protein kinase produces an autonomous enzyme in Aplysia neurons. AB - We have shown previously that the subcellular distribution of a major calmodulin binding protein is altered under conditions causing increased synthesis of cAMP in Aplysia neurons (Saitoh, T., and J. H. Schwartz, 1983, Proc. Natl. Acad. Sci. USA, 80:6708-6712). We now provide evidence that this Mr 55,000 protein is a subunit of a Ca2+/calmodulin-dependent kinase: (a) both the Mr 55,000 calmodulin binding protein and kinase activity are loosely attached to the membrane cytoskeletal complex; (b) both kinase activity and the Mr 55,000 protein are translocated from the membrane-cytoskeleton complex to the cytoplasm under conditions that cause the change in the subcellular distribution of the Mr 55,000 calmodulin-binding protein; and (c) calmodulin-binding activity of the Mr 55,000 protein and the ability to carry out the Ca2+/calmodulin-dependent phosphorylation of synapsin I are purified in parallel. The subcellular localization of the Ca2+/calmodulin-dependent protein kinase appears to be under control of two second messengers: Ca2+ and cAMP. We find that the Mr 55,000 subunit is phosphorylated when the extracted membrane-cytoskeleton complex is incubated with Ca2+, calmodulin, and ATP, with the concomitant release of this phosphorylated peptide from the complex. Previously, we had found that, when translocation occurs in extracts in the presence of cAMP and ATP (but in the absence of Ca2+), there was no detectable phosphorylation of the Mr 55,000 subunit itself. The subcellular distribution of the subunit thus appears to be influenced by (a) cAMP-dependent phosphorylation, which, we infer, modifies some as yet unidentified structural component, causing the release of the enzyme; and (b) Ca2+/calmodulin-dependent phosphorylation of the Mr 55,000 subunit. These studies also suggest that phosphorylation has an important regulatory consequence: during the Ca2+/calmodulin-dependent translocation of the Mr 55,000 subunit, the kinase appears to be activated, becoming independent of added Ca2+/calmodulin. PMID- 2982888 TI - Calcium regulation of phosphatidyl inositol turnover in macrophage activation by formyl peptides. AB - Stimulation by the tripeptide N-formyl norleucyl leucyl phenylalanine (FNLLP) of the guinea pig alveolar macrophage gives rise to transient production of superoxide anion (O2-). Components of the phosphatidyl inositol (PI) cycle (phosphatidic acid (PA), phosphatidyl inositol-4,5-bisphosphate (TPI) and phosphatidyl inositol-4-phosphate (DPI) were monitored using 32P in order to examine the possible association of this cycle with the FNLLP-stimulated production of O2-. Macrophage stimulation by FNLLP led to an increased flux of metabolites through the PI cycle. The level of 32P label in both TPI and DPI rapidly decreased upon exposure to FNLLP, followed by a 5-min period during which the 32P label in TPI and DPI approached prestimulated levels. During this period, there was a fivefold increase in 32P-PA. It is suggested that diacylglycerol (DAG) is the O2- -activating intermediate in the stimulated mechanism, as evidenced by the buildup of PA (for which DAG is the precursor) in parallel with the time course of O2- production. The importance of continued cycling of PI in the stimulated mechanism is demonstrated by the inhibition by LiCl of the extent, but not the initial rate, of both O2- production and the formation of 32P-PA upon peptide stimulation after 1-h preincubation with 10 mM LiCl. The influence of calcium on this mechanism was also examined. It has previously been demonstrated that intracellular availability of calcium can influence the rate and extent of O2- production. In cells preloaded with quin-2, which acts as a high-affinity sink for calcium in the cytosol, the initial rate of FNLLP-stimulated O2- production is inhibited in low (10 microM) extracellular calcium medium. High extracellular calcium (1 mM) completely reverses this inhibition and also significantly extends the time course of O2- production in both quin-2 and control cells (Stickle et al., 1984). In parallel with these effects on O2- production, varying calcium conditions is demonstrated to influence the rate and extent of PA formation. These same calcium conditions were found to have little or no effect on the initial unstimulated levels of TPI, DPI, and PA. These results indicate that the influence of an intracellular pool of calcium on O2- production may be via its influence on stimulated PI turnover. PMID- 2982889 TI - The effect of ionophores and related agents on the induction of doming in a rat mammary epithelial cell line. AB - Addition of dimethyl sulfoxide (DMSO) and the mammotropic hormones prolactin, hydrocortisone, insulin, and estradiol to confluent cultures of the epithelial cell line Rat Mammary (Rama) 25 increases dramatically the formation of domes in the cell monolayer after 48-72 hr. Associated with the increase in doming is an increase of 24% in the activity of the Na+/K+ ATPase. Both Ca2+ (A23187) and Na+ (monensin, gramicidin J, melittin) ionophores can replace DMSO in inducing domes, whilst the K+ ionophore valinomycin inhibits doming. However, there are no synergistic nor additive effects, respectively, with suboptimal or optimal concentrations of A23187 and melittin together. Ouabain, at concentrations which inhibit the Na/K ATPase in vitro, and amiloride, at concentrations reported to inhibit the passive transport of Na+, both inhibit completely the formation of domes induced by DMSO, A23187, and melittin. EGTA, however, inhibits only the induction of doming by DMSO and A23187; it is without effect with melittin. A23187 and melittin induce the major polypeptide changes that occur in doming cultures with DMSO, and most of these changes are also inhibited with ouabain. It is suggested that one possible interpretation of the findings is that the induction of doming by DMSO in Rama 25 cells occurs by means of sequential increases in Ca2+ and Na+ influxes into the cell, and that the increased intracellular concentration of Na+ so produced stimulates the Na+/K+ ATPase, with a net effect of pumping liquid beneath the cellular monolayer. PMID- 2982890 TI - Specific ouabain binding to brain microvessels and choroid plexus. AB - The energy-dependent transport of ions across the blood-brain barrier and the blood-cerebrospinal fluid barrier by Na+, K+-ATPase is credited with an important role in brain homeostasis. In this study, we have assessed the relative enrichment of Na+, K+-ATPase in regional brain capillary preparations and in the choroid plexus by the quantitative determination of the cardiac glycoside binding sites in these preparations using [3H]ouabain as a ligand. We find that ouabain binds specifically to brain microvessels of the rat and the pig and to the choroid plexus of the pig in a saturable manner. The maximum density of ouabain binding sites in brain microvessels of both species is about one-fourth that of the crude membranes of the cerebrum and cerebellum. The density of ouabain binding sites in the pig choroid plexus is intermediate between that of the brain and brain microvessels. We do not find regional differences in ouabain binding to membrane fractions of the cerebrum and cerebellum, nor any significant differences in ouabain binding to cerebral and cerebellar microvessels. These findings provide quantitative estimates of Na+, K+-ATPase in brain capillaries and choroid plexus. PMID- 2982892 TI - Radioimmunoassay for detection of VP1 specific neutralizing antibodies of foot and mouth disease virus. AB - A solid-phase radioimmunoassay was developed for the detection of antibodies against a specific region of the VP1 protein of the A24 and 01 serotypes of foot and mouth disease virus. The antibody titers from the radioimmunoassay showed a positive correlation with neutralizing antibody titers determined by a mouse protection assay. The specificity of the assay resides in the peptide used as antigen. The assay is rapid, reproducible and does not require the use of whole virions. PMID- 2982891 TI - A combined enzyme immunoassay for rotavirus and adenovirus (EIARA). AB - A combined enzyme immunoassay for rotavirus and adenovirus (EIARA) was developed as a double-antibody sandwich assay in which test samples are added to plastic wells coated with rotavirus or adenovirus goat antibody. The presence of antigens is detected by mixed-guinea pig antisera to the same viruses followed by rabbit anti-guinea pig IgG conjugated with peroxidase and subsequently by ortho phenylenediamine substrate. Titrations of rotavirus (SA11) and adenovirus type 2 tissue culture grown viruses in the presence of separate capture sera and mixed detector sera revealed that the two virus-antibody reactions occurred independently and were not affected by each other. Comparison with another enzyme immunoassay for rotavirus, with immunoelectron microscopy and with polyacrylamide gel electrophoresis showed that EIARA is a sensitive and specific method for detecting rotaviruses and adenoviruses in faeces from children with gastroenteritis. PMID- 2982893 TI - Detecting bluetongue virus RNA in cell culture by dot hybridization with a cloned genetic probe. AB - A 70% copy of genome segment 7 of bluetongue virus (BTV)-17 has been cloned into the plasmid pBR-322. This cloned BTV segment when used as a radioactive probe will hybridize to BTV double-stranded RNA extracted from cell cultures and dotted onto nitrocellulose paper. This dot hybridization technique is therefore suitable for detecting and identifying BTV in cell culture. The specificity of cloned probes is discussed in relation to detecting gene sequences specific for either the bluetongue serogroup or different serotypes of BTV. PMID- 2982894 TI - Health registry and clinical data base technology: with special emphasis on cancer registries. PMID- 2982895 TI - 17-Hydroxyprogesterone responses to adrenocorticotropin in children with premature adrenarche. AB - The adrenal secretory response to an iv bolus dose of ACTH was measured in 10 girls (4-8 yr of age), 5 boys (4-9 yr) with premature adrenarche (PA), and 20 normal children. The evening before the ACTH test, each subject took dexamethasone (1 mg at bedtime) to suppress the early morning surge of ACTH. The next morning, 2 serum samples were obtained before the administration of ACTH (Cortrosyn; 0.25 mg), and 2 samples were collected 30 and 45 min after ACTH administration. All samples were assayed for cortisol, dehydroepiandrosterone (DHEA), DHEA sulfate, 17-hydroxyprogesterone (17-OHP), and androstenedione. There was no significant difference in the dexamethasone-suppressed steroid levels between the children with PA and the normal children. After ACTH injection, cortisol, DHEA, 17-OHP, and androstenedione increased significantly. There was no significant change in DHEA sulfate. The mean 17-OHP response to ACTH in girls with PA was significantly higher than that in girls and women whose pubertal development was normal. This response was similar in magnitude to that in a group (n = 5) of obligate heterozygotes for congenital adrenal hyperplasia (CAH). These data suggest that many girls with PA have a mild adrenal steroid secretory defect that resembles the response in adult obligate heterozygotes for CAH due to 21 hydroxylase deficiency. In contrast to the girls, none of the boys with PA had an exaggerated 17-OHP response to ACTH. Thus, these boys had no evidence for an adrenal steroid secretory defect. In summary, although the clinical presentation of boys with PA is similar to that of girls with PA, there is a significant difference in the adrenal steroid secretory response to ACTH. Thus, the biochemical events that cause PA in boys may be different from the corresponding events in girls. PMID- 2982896 TI - Late-onset adrenal steroid 3 beta-hydroxysteroid dehydrogenase deficiency. I. A cause of hirsutism in pubertal and postpubertal women. AB - To investigate the adrenal cause of hyperandrogenism in peri- and postpubertal hirsute women, baseline and ACTH-stimulated serum concentrations of delta 5-17 hydroxypregnenolone (delta 5-17P), dehydroepiandrosterone (DHEA) and its sulfate, 17-hydroxyprogesterone (17-OHP), cortisol, delta 4-androstenedione, and testosterone were determined in 116 women with hirsutism or acne of peri- and postpubertal onset with or without menstrual abnormalities. The results were compared with the same steroid concentrations in 30 normal age-matched women. Sixteen of the 116 women with hirsutism whose ACTH-stimulated 17-OHP levels (mean +/- SD, 5404 +/- 3234 ng/dl; normal, 334 +/- 194) were markedly elevated while their ratios of delta 5-17P to 17-OHP (0.4 +/- 0.2; normal, 3.4 +/- 1.5) were low were diagnosed as having nonclassical symptomatic 21-hydroxylase deficiency. Seventeen other hirsute women, including 3 siblings, had very high responses of delta 5-17P (2276 +/- 669 ng/dl; normal, 985 +/- 327) and DHEA (2787 +/- 386 ng/dl; normal, 1050 +/- 384) to ACTH stimulation, with significantly elevated ratios of delta 5-17P to 17-OHP (11 +/- 2.0; normal, 3.4 +/- 1.5) and DHEA to delta 4-androstenedione (7.5 +/- 2.3; normal, 4.6 +/- 1.5). In these hirsute women, the morning serum delta 5-17P and DHEA concentrations were elevated, had a diurnal variation, and were suppressed with dexamethasone administration. We propose that partial adrenal 3 beta-hydroxysteroid dehydrogenase deficiency is the cause of hirsutism in these women. This may represent an allelic variant at the genetic locus for 3 beta-hydroxysteroid dehydrogenase deficiency similar to that reported for symptomatic nonclassical 21-hydroxylase deficiency producing peripubertal excess androgen syndrome. PMID- 2982897 TI - Characterization of thyroxine-binding globulin secreted by a human hepatoma cell line. AB - T4-binding globulin (TBG) is a glycoprotein synthesized by the liver and is the principal carrier of T4 and T3 in serum. In this report, we demonstrate that the Hep G2 cell line, derived from a human hepatoblastoma, synthesizes and secretes TBG, the properties of which were characterized. Hep G2 cells secreted TBG into the medium after more than 100 transfers in tissue culture conditions. At confluency and after changing to serum-free culture conditions, TBG accumulation into the medium was linear for 3 days and constituted approximately 0.16% of the proteins synthesized over 24 h. Its abundance relative to albumin is 10-fold greater than that found in normal human serum. TBG secreted by the Hep G2 cells was indistinguishable from native normal human serum TBG, as determined immunologically, by electrophoresis on polyacrylamide gel in denaturing and nondenaturing conditions, and by isoelectric focusing. It also specifically bound T4 and T3, albeit with slightly reduced affinity, and had increased heat lability. Although slightly different from normal serum TBG in caucasians, the physical and biological properties of the Hep G2-derived TBG are similar to those of the variant TBG found in the serum of some healthy Australian Aborigines. PMID- 2982898 TI - Vitamin D resistance and alopecia: a kindred with normal 1,25-dihydroxyvitamin D binding, but decreased receptor affinity for deoxyribonucleic acid. AB - A new kindred exhibiting vitamin D resistance and alopecia is described. Clinically, three of seven sisters demonstrated rickets, hypocalcemia, elevated serum 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] levels, and alopecia. Biochemical analysis of cultured fibroblasts from skin biopsy explants in two affected and one normal sister revealed normal [3H]1,25-(OH)2D3 binding to receptors (Kd = 0.05 nM; Nmax = 30-50 fmol/mg protein). Despite normal steroid binding, cells from the two affected sisters failed to respond to 1,25-(OH)2D3 in vitro, as measured by induction of the enzyme 25-hydroxyvitamin D-24-hydroxylase. The cells from the normal sister showed a response within the range of five normal cell lines. Sucrose gradient analysis yielded a typical 3.2S protein under high salt conditions in extracts from the three siblings, but with reduced capacity to aggregate to a 6S moiety in low salt gradients in the two affected cells. Whole cell [3H]1,25-(OH)2D3 binding studies revealed nearly normal localization of bound receptor to the nuclear compartment. Elution of bound receptors by KCl gradients from both DNA-cellulose or fibroblast nuclei demonstrated that the receptors from the affected sisters exhibited decreased affinity for DNA compared to those from normal subjects. We conclude that 1,25-(OH)2D3 receptors from these resistant fibroblasts have a normal steroid-binding domain, but a defective nuclear binding domain. We believe that this abnormality may be responsible for the vitamin D resistance observed both in vivo and in vitro. PMID- 2982899 TI - Cushing's syndrome due to ectopic production of corticotropin-releasing factor. AB - A patient with Cushing's syndrome is described who had a metastatic medullary carcinoma of the thyroid which contained corticotropin-releasing factor. ACTH was found by an immunohistochemical method in the patient's pituitary, but not in the thyroid tumor. This is the second report demonstrating corticotropin-releasing factor in tumor tissue in this syndrome. Wider use of immunohistological methods can help distinguish this variety from other tumors associated with the ectopic ACTH syndrome when bioassays are not available. PMID- 2982900 TI - The effect of dexamethasone on the 24-hour profiles of adrenocorticotropin and cortisol in Cushing's syndrome. AB - ACTH and cortisol are normally secreted episodically rather than continuously. This characteristic of episodic secretion is preserved in patients with Cushing's syndrome. To determine whether exogenous glucocorticoids modulate this pulsatility and to study its possible etiological implications, we obtained 24-h plasma cortisol profiles in seven patients with Cushing's syndrome (five Cushing's disease, one adrenal adenoma, and one bilateral adrenal cortical macronodular hyperplasia) before and during suppression with various doses of dexamethasone [low (0.5 mg, every 6 h), high (2 mg, every 6 h), and very high (4 mg, every 6 h)]. Simultaneous 24-h plasma ACTH profiles were obtained in two patients with Cushing's disease. Blood was drawn at 30-min intervals for 25 h. Individual profiles were analyzed to determine the 24-h mean level, the presence of a circadian component and its amplitude, and the number and magnitude of significant secretory pulses over the 24-h span. The concordance between significant ACTH and cortisol pulses also was quantified. Baseline values in patients were compared to those in seven normal subjects. Under basal conditions, the 24-h mean cortisol level was 3- to 4-fold higher than normal in all patients with Cushing's syndrome. In contrast, the basal 24-h mean ACTH level was normal in one, and slightly elevated in the other of the two patients with Cushing's disease in whom plasma ACTH concentrations were measured. However, in contrast to the normal subjects, all ACTH values were above 10 pg/ml even during the period of minimal secretion. Basal circadian variation in adrenocortical activity, albeit of reduced amplitude, was found in four of five patients with Cushing's disease; it was absent in the patient with adrenal adenoma. Low dose dexamethasone reduced the 24-h mean cortisol level and increased the amplitude of the circadian rhythm, unmasking a diurnal rhythm in the single patient with Cushing's disease in whom no significant circadian periodicity was present in the basal condition. This effect was further increased with the high dose of dexamethasone, which concomitantly reduced the number and increments of the secretory pulses. A lesser effect was found in the patient with bilateral nodular hyperplasia, and no effect was seen in the patient with adrenal adenoma. ACTH pulsatility, but not diurnal rhythm, also was dampened by dexamethasone. Reduction in the magnitude, but not the number, of ACTH secretory pulses by dexamethasone produced a reduced concordance ratio of ACTH with cortisol pulses of 0.39, compared to 0.64 in the basal state.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982901 TI - The effects of radiographic contrast agents and other compounds on the nuclear binding of L-[125I]triiodothyronine in dispersed human skin fibroblasts. AB - Using a dispersed intact cell assay system, we screened a number of compounds for their ability to compete for nuclear binding of L-[125I]T3 in cultured human skin fibroblasts incubated for 90 min at 37 degrees C. T3 inhibited nuclear [125I]T3 binding by 50% at a concentration of 3.4 +/- 0.3 (+/- SE) X 10(-10) M. 3,5 Dimethyl-3'-isopropyl-thyronine, a nonhalogenated thyroid hormone agonist, had an affinity for the nuclear thyroid hormone receptor (4.4 X 10(-9) M, as judged by 50% inhibition of nuclear [125I]T3 binding) that correlates well with its thyromimetic potency. Of several radiographic contrast agents and other compounds tested (iodipamide, iopanoic acid, sodium ipodate, sodium diatrizoate, sodium tyropanoate, diphenylhydantoin, carbamazepine, amiodarone hydrochloride, propylthiouracil, propranolol, and potassium iodide), only sodium ipodate (Oragrafin) and iopanoic acid (Telepaque) interfered with nuclear [125I]T3 binding, with 50% inhibition at 5 X 10(-5) and 1.8 X 10(-4) M, respectively. Interestingly, diphenylhydantoin and amiodarone, two compounds previously thought to interact with thyroid hormone receptors, did not impair fibroblast nuclear [125I]T3 binding at concentrations up to 10(-3) M. We conclude that this in vitro assay system with intact human cells is useful in evaluating the nuclear T3 receptor affinity of compounds that affect thyroid hormone action or metabolism. These studies more closely approximate in vivo conditions and, therefore, provide information not obtainable by studies with isolated nuclei or nuclear extracts. PMID- 2982902 TI - Dexamethasone preparation does not alter corticoid and androgen responses to adrenocorticotropin. AB - ACTH tests were performed with and without dexamethasone (dex) pretreatment to clarify the nature of the relationship between the absolute and incremental response (delta) to ACTH in normal men and women, hirsute women, adrenarchal children, and women heterozygous for congenital adrenal hyperplasia (CAH). The purposes were to test the effect of dex preparation on adrenal responsiveness to ACTH and the efficacy of the dex-pretreated ACTH test in detecting heterozygosity for CAH. Cosyntropin was given as a 10 micrograms/m2 iv bolus dose at 0800-1000 h; dex (1 mg/m2) was given at 2200-2400 h the night before. Dex did not alter the absolute plasma steroid levels achieved in response to ACTH. However, since post dex baseline concentrations of adrenal steroids were lower, the delta to ACTH was significantly greater for the major adrenal secretory products, 17 alpha hydroxypregnenolone (3 beta, 17 alpha-dihydroxypregn-5-ene-20-one), dehydroepiandrosterone, and cortisol (F). For example, for all paired tests, the mean plasma F values achieved 30 min post-ACTH were 26.0 +/- 4.4 (+/- SD) micrograms/dl without dex and 23.8 +/- 5.5 micrograms/dl after dex. In contrast, the mean delta of plasma F 30 min post-ACTH was less without (13.3 +/- 4.8 micrograms/dl) than after (19.4 +/- 3.3 micrograms/dl) dex (P less than 0.001). Apparent 21-hydroxylase efficiency, computed from dex-prepared tests, was found in follicular phase women to have a markedly skewed distribution without clear demarcation between 15% of the population and CAH heterozygotes. Luteal phase responses differed from follicular phase responses in dex-pretreated women in the magnitude of the 17-hydroxyprogesterone response. In the luteal phase, although the plasma 17-hydroxyprogesterone level at 30 min was higher, a response to ACTH was not consistently found, averaging only 22 +/- 26 ng/dl, in contrast to the consistent 52 +/- 15 ng/dl response in the follicular phase. These findings have practical implications for interpreting rapid ACTH test results. The absolute plasma F level achieved post-ACTH is more important as an index of adrenocortical reserve than the increment. Dex pretreatment appears to offer no practical advantage in ACTH testing for mild defects in 21-hydroxylation; we postulate that this is because of considerable normal variability in the efficiency of 21 hydroxylation. PMID- 2982903 TI - Rapid sequential intravenous administration of four hypothalamic releasing hormones as a combined anterior pituitary function test in normal subjects. AB - Normal subjects were studied to test the feasibility of a combined anterior pituitary function test using iv administration of four hypothalamic releasing hormones: ovine corticotropin-releasing hormone, human GH-releasing hormone, GnRH, and TRH. Initially, nine normal men were studied with various combinations of these four hormones to exclude the possibility that they might inhibit or synergize with each other in releasing the individual anterior pituitary hormones. When given in combination, the releasing hormones were administered as sequential 20-sec iv infusions in the following order and doses: ovine corticotropin-releasing hormone, 1 microgram/kg; GnRH, 100 micrograms; human GH releasing hormone, 1 microgram/kg; and TRH, 200 micrograms. Plasma or serum samples were assayed for ACTH, cortisol, GH, PRL, FSH, LH, and TSH at multiple times for 120 min after injection. Compared to individual administration, combined administration of these four hypothalamic releasing hormones caused no apparent inhibition or synergism with respect to the individual hormone responses of these normal subjects. Side-effects of the combined test were the same as those observed with individual hormone administration. No unusual or dangerous side-effects were observed. Having confirmed the efficacy of combined administration of the four releasing hormones, we administered the combination to five additional normal men and 12 normal women. Anterior pituitary hormone and cortisol responses were the same in men and women, except for a lower LH and a greater PRL response in women. There was a rapid increase in all hormones, with peak levels usually reached by 60 min. Adequate assessment of individual hormone responses can be achieved by assaying a basal and only 2 (or 3 in the case of ACTH and GH) postinfusion samples. A rapid, safe, and useful test of combined anterior pituitary function appears to be feasible using these four hypothalamic releasing hormones. PMID- 2982904 TI - Combined 21- and 11 beta-hydroxylase deficiency in familial congenital adrenal hyperplasia. AB - Studies in three families (A, B, and C) revealed five patients with congenital adrenal hyperplasia (CAH) due to partial and combined 21- and 11 beta-hydroxylase deficiency. One patient (A-11 1), a 23-yr-old severely virilized chromosomal female, was reared as a male, and two females (B-11 2 and C-1) complained only of hirsutism, acne, and menstrual abnormalities. Patients A-11 2 and B-11 8 (17 1/2 and 10 yr old) were asymptomatic and detected by finding an HLA genotype identical to that of their respectively affected brother and sister. Three patients (A-11 1, A-11 2, and C-1) had moderate hypertension. In spite of the wide range of clinical manifestations, all individuals had elevated androgen levels, while cortisol secretion was severely impaired only in A-11 2. 21 Hydroxylase deficiency was diagnosed on the basis of markedly increased plasma and urinary levels of 17-hydroxyprogesterone (17-OHP) and 21-deoxycortisol and their respective urinary metabolites pregnanetriol and pregnanetriolone. PRA was elevated in three patients, while urinary aldosterone was normal or increased. 11 beta-Hydroxylase deficiency was diagnosed on the basis of increased 11 deoxycortisol and deoxycorticosterone in plasma and tetrahydro-11-deoxycortisol and deoxycorticosterone in urine, particularly after ACTH administration. In contrast to classical 11 beta-hydroxylase deficiency CAH, urinary 18 hydroxycorticosterone and 18-hydroxy-11-deoxycorticosterone were normal or elevated. The nature and mechanism of a combined enzymatic defect are unknown. The coincidental presence in a single individual of the mutant genes for both 21- and 11 beta-hydroxylase deficiency CAH is very unlikely to occur. Two alternative hypotheses may explain our findings. One is the existence of a genetically inherited abnormal (or aberrant) 11 beta-hydroxylase, whose affinity for its normal substrate is changed for an abnormal one (17-OHP). As a result, 11 beta hydroxylation of 11-deoxycortisol is deficient while 17-OHP 11 beta-hydroxylation is markedly enhanced. Thus, both 11-deoxycortisol and 21-deoxycortisol as well as their urinary metabolites accumulate. The ability for 18-hydroxylation, however, remains normal. In this case, 21-hydroxylase is not deficient, yet 21 deoxycortisol cannot be further hydroxylated to cortisol, since this steroid is not a suitable substrate for the enzyme. Such a disorder may represent a new allelic variant of 11 beta-hydroxylase deficiency CAH, which, similar to 21 hydroxylase deficiency, is completely linked to the HLA complex.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2982905 TI - Adenylate cyclase activity in neocortex and fetal zone membrane fractions of the human fetal adrenal gland. AB - Whether peptide hormones other than ACTH may be responsible for the difference in size or rate and pattern of steroidogenesis of the fetal zone (FZ) compared to those of the neocortex (NC) of the human fetal adrenal gland is controversial. In the present investigation, the activity of adenylate cyclase in membrane fractions of separated zones of the human fetal adrenal gland was determined. Basal adenylate cyclase activity was 2- to 3-fold greater in NC than in FZ membrane fractions. The addition of ACTH-(1-24) stimulated adenylate cyclase activity in both zones, but the activity was more sensitive to ACTH (10(-10) M) in NC fractions than in FZ fractions (10(-7) M). In addition to ACTH-(1-24), the effect of other ACTH-related peptides on the activity of adenylate cyclase in the separated zones of the adrenal gland was investigated. 16K fragments 2-36, gamma 3MSH, alpha MSH, beta-endorphin, leu-enkephalin, and met-enkephalin, as well as hCG, FSH, prostaglandin E2, prostaglandin F2 alpha, epinephrine, and norepinephrine did not stimulate adenylate cyclase activity in either zone. It is concluded that basal and ACTH-(1-24)-stimulated adenylate cyclase activities are greater in NC than in FZ membrane fractions. In addition, the results of the present investigation do not support the concept that other ACTH-related peptides or peptide or protein hormones increase steroidogenesis by stimulating adenylate cyclase activity in the human fetal adrenal gland. PMID- 2982906 TI - Specific growth hormone receptors on human peripheral mononuclear cells: reexpression, identification, and characterization. AB - Although specific GH receptors have been demonstrated in various tissues of a number of species, the presence of GH receptors on human peripheral mononuclear cells (PMC) is controversial. Binding of human GH (hGH) to its receptor as the hypothesized initial step of hormone action was consequently studied using mononuclear cells from peripheral venous blood of normal subjects. Specific binding of [125I]hGH was rapid, reversible, and time and temperature dependent. Specific GH binding to PMC was maximal after 8-24 h of preincubation. Binding of hormone was maximal at 37 C after incubation of cells for 2 h. Dissociation of GH was maximal at 37 C after the addition of 6 M NaCl. A linear relationship between specific GH binding and cell number was found. Saturation of GH binding to 10(6) PMC was obtained with 25 ng iodinated hormones. Half-maximal inhibition of GH binding occurred at 12-25 ng unlabeled hGH/tube. Hypothalamic and pituitary hormones as well as insulin did not interfere with specific hGH binding to PMC. Scatchard analysis of [125I]hGH binding to PMC revealed a receptor with a mean affinity constant of 1.5 +/- 0.2 (+/- SD) X 10(9)/M-1 (n = 72) and a maximal binding capacity of 7.1 +/- 2.0 X 10(-11) M/10(6) cells. The concentrations of calcium, sodium, and magnesium ions in the incubation medium strongly influenced GH binding, whereas pH or potassium concentration did not. As interassay variation of the binding assay was low (14% for total binding; 6% for specific hGH binding), this direct approach to study tissue receptors for hGH in a human in vitro test was reproducible and should encourage the investigation of receptor regulation as well as the study of binding in human disease. PMID- 2982907 TI - Genetic differences between the salt-wasting, simple virilizing, and nonclassical types of congenital adrenal hyperplasia. AB - Human leukocyte antigen (HLA) alleles and plasma 17-hydroxyprogesterone levels after ACTH stimulation were studied in 134 German families of patients with the salt-wasting (SW), simple virilizing (SV), or nonclassical (NC) late-onset form of congenital adrenal hyperplasia (CAH). Unexpected hormonal evidence for CAH was found in 6 otherwise healthy members of the relatives' group, who, therefore, were considered to be NC cryptic cases. HLA typing revealed a genetic difference between the 2 classical disease forms; SW CAH was strongly associated with Bw47, whereas SV CAH was closely linked to B5(w51). It also confirmed the nearly complete connection of NC CAH with B14. These alleles, especially Bw47 and B14, are mostly components of normally rare haplotypes: A3,Bw47,DR7 and Aw33,B14,DR1, respectively. They do not occur in the families' disease-unaffected haplotypes. Thus, it may be that all or almost all individuals from the general population bearing 1 of them are in fact CAH heterozygotes. Moreover, it seems possible to predict the severity of an infant's disease from his genomic type. The HLA linkage data were consistent with those obtained from ACTH testing, which showed significantly higher 17-hydroxyprogesterone increases in the genetically defined heterozygous relatives of SW patients than in the respective members of SV families. Of the families, 2 were also informative for mapping of the CAH disease gene(s) within the HLA-B to Glo interval. PMID- 2982908 TI - Hypothalamic dysfunction in overtrained athletes. AB - Some athletes who undertake strenuous training programs for a prolonged period of time develop the overtraining syndrome. The pathophysiology of the condition is unknown. Hypothalamic-pituitary function was studied by determining the hormonal responses to insulin-induced hypoglycemia in five asymptomatic male marathon runners during a 4-month period in which they ran 42-, 56-, and 92-km races and in four overtrained male athletes. The response of the asymptomatic runners was not different when tested 1 month before and within 48 h after the 42- and 92-km races. All four overtrained athletes presented with impaired training and racing times, apathy, and a heavy-legged feeling and were tested when overtrained and again after 4 weeks of rest. The plasma cortisol, ACTH, GH, and PRL responses to insulin-induced hypoglycemia in the four overtrained athletes were lower than their responses after the rest and lower than the responses of the asymptomatic runners. In both groups, the LH, TSH, and PRL responses to LHRH and TRH were normal. The impaired hormonal responses to insulin-induced hypoglycemia, with recovery after 4 weeks of rest, indicate hypothalamic dysfunction and may be a diagnostic marker of the overtraining syndrome. PMID- 2982909 TI - [A seroepidemiological study of polio in Thailand and investigation of the OPV (oral polio vaccine) immunization]. PMID- 2982910 TI - Use of a sodium polyanetholesulfonate disk for the identification of Gardnerella vaginalis. AB - Several methods have been previously suggested for the presumptive identification of Gardnerella vaginalis in clinical laboratories, but none is entirely satisfactory. We previously found that sodium polyanetholesulfonate (SPS) inhibits G. vaginalis in blood culture media. We compared susceptibility to an SPS-containing paper disk with beta-hemolysis on human blood agar, hippurate hydrolysis, and inhibition by alpha-hemolytic streptococci for identification of 62 previously confirmed G. vaginalis strains. All strains were positive by SPS disk and alpha-hemolytic streptococcus inhibition, 78% were positive by beta hemolysis, and 81% were positive by hippurate hydrolysis. Although positive reactions occurred with SPS disk and alpha-hemolytic streptococcus tests for 5 and 9 of 84 other bacteria tested, respectively, none of these bacteria were positive for both tests. We conclude that a combination of SPS disk susceptibility and alpha-hemolytic streptococcus inhibition provides excellent identification of G. vaginalis when performed by the methods suggested. PMID- 2982911 TI - Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens. AB - A monoclonal antibody was used to detect an early antigen of cytomegalovirus (CMV) by fluorescence 16 h after inoculation of MRC-5 monolayers in 1-dram (ca. 3.7-ml) shell vials and low-speed centrifugation. Of 770 specimens (urine, blood, lung tissue, sputum) processed in shell vials, 124 (16%) were positive for the virus at 16 h postinfection. CMV was isolated in standard tube cell cultures (average time, 9 days) from only 88 specimens, but there were no instances (with the exception of 2 blood specimens) in which CMV was recovered from tube cultures but not from shell vials. Additional specimens from 18 patients were positive in the shell vial assay but negative in the conventional tube cell culture assay. Other specimens from 14 of the 18 patients yielded CMV in conventional tube cell cultures. Of the 4 patients from whom CMV was not recovered from other specimens by conventional tube cell culturing, all had evidence of recent CMV infections, as indicated by a fourfold or greater rise in antibody titer. The specificity of the shell vial assay for the detection of CMV is supported by assays of other specimens from the same patients yielding the virus or serological evidence indicating recent infections, the known enhancement of CMV detection after centrifugation of the shell vials, and the distinct and easily recognizable fluorescence confined to the nuclei of CMV-infected cells. Our data indicate that the shell vial cell culture assay for the detection of CMV is as specific as and more sensitive than conventional tube cell culturing for the diagnosis of CMV infections. PMID- 2982912 TI - Adherent cells suppress measles and herpes simplex I virus-induced blastogenesis of multiple sclerosis lymphocytes. AB - Viral antigen-induced blastogenesis of lymphocytes from multiple sclerosis (MS) patients was investigated to determine if the responses were actively suppressed. We found that depletion of adherent cells increased measles and herpes simplex I virus antigen-induced transformation of MS lymphocytes. Addition of indomethacin to cultures of unfractionated MS lymphocytes also caused an increase in viral antigen-induced responses. These two facts, plus finding that the cell type mediating the immunosuppression did not rosette with 2-aminoethylisothiouronium bromide hydrobromide-treated sheep red blood cells, indicate that the suppressed T-cell responsiveness of MS patients is caused by macrophages rather than T cells. These results have a major implication for the divergent published data on blastogenesis induced in MS patient lymphocytes by specific antigens, viral or otherwise. We feel the inconsistencies may simply have arisen from the different lymphocyte isolation and washing procedures used giving variable levels of macrophages and, hence, variable levels of immune suppression. This clearly suggests that induction of blastogenesis in MS patient lymphocytes by a wider array of infectious agent antigens and by various neural antigens should now be undertaken using adherent cell-depleted lymphocytes. PMID- 2982913 TI - Glucagon receptor of human liver. Studies of its molecular weight and binding properties, and its ability to activate hepatic adenylyl cyclase of non-obese and obese subjects. AB - The glucagon receptor and the adenylyl cyclase system of human liver membranes were studied in six non-obese and six obese subjects who had elevated insulin and plasma glucagon levels. Analysis of specific glucagon binding by the method of Scatchard demonstrated a linear (monocomponent) plot with a dissociation constant of 2-3 nM, and the binding at low hormone concentrations was sensitive to guanosine triphosphate (GTP). The molecular weight of the glucagon receptor was 63,000 D as determined by an affinity labeling procedure and sodium dodecyl sulfate gel electrophoresis. Affinity labeling of this structure was specific for glucagon and inhibited by GTP. Glucagon stimulated the production of cyclic adenosine monophosphate (cAMP) by human membranes with half-maximal activation elicited by 6 nM hormone. The human cyclase system required GTP to facilitate an optimal glucagon response. NaF (10 mM) also activated the cyclase system and produced the same magnitude of response as maximum glucagon activation. A comparison of the liver adenylyl cyclase system of non-obese and obese subjects was made using glucagon (5 nM and 1 microM) and NaF (10 mM). No significant differences in cAMP production were noted between the two groups, regardless of the agent used to activate the enzyme. These findings agree with the glucagon binding studies that showed similar amounts of binding activity in the membranes from the two groups. Also, there was no influence of either age or sex of the subjects on the adenylyl cyclase response. In conclusion, human liver membranes contain a glucagon receptor and an adenylyl cyclase system that correspond closely to the well-studied system in animal liver. This system in human obesity is not altered by the approximately twofold elevation in plasma glucagon that occurs in this metabolic disorder. PMID- 2982914 TI - Studies of human C5a as a mediator of inflammation in normal human skin. AB - C5a is an 11,000-D fragment of the fifth component of complement (C5) with potent anaphylatoxic and leukocyte chemotactic activities. C5a is believed to play an important role in the pathophysiology of certain skin disorders and systemic diseases with cutaneous manifestations. However, there is very little known about the in vivo reactivity of C5a in man. In this study we examined the effects of intradermal injections of human C5a in 17 normal volunteers. C5a was prepared by interacting highly purified human C5 with zymosan bound alternative pathway C5 convertase under conditions resulting in consumption of approximately 90% of the C5 substrate. C5a produced in this manner was chemotactic for human neutrophils and monocytes (0.5 X 10(-7) to 10(-9) M) and caused neutrophil aggregation and myeloperoxidase release (concentrations greater than or equal to 10(-10) M) in vitro. In vivo, C5a produced immediate wheal and flare reactions in all volunteers, and was active in doses as low as 1 ng (10(-13) mol). Intradermal testing with 20 ng of C5a in eight volunteers produced a maximal mean wheal of 11.75 mm (+/- 0.80 mm SEM) 20 min after anaphylatoxin injection, and a maximal mean erythema of 62.50 mm (+/- 3.27 mm SEM) 10 min after C5a administration. Reactions at C5a test sites were dose-related, associated with marked pruritus in some subjects, resolved without lesion formation, and were not associated with late phase reactions. In vivo testing revealed that human C5a was a more potent mediator of wheal and flare reactions than histamine, 48/80, human C3a, or morphine sulfate. Skin biopsies from eight volunteers 20 min after intradermal injection of 20 ng of C5a revealed a neutrophil-predominant perivascular infiltrate, endothelial cell edema, and sites of leukocytoclasis. Mast cell degranulation was observed on both light and electron microscopy of biopsies from C5a test sites. Although erythema at C5a injection sites was reduced by pretreating volunteers with hydroxyzine, whealing reactions and cellular infiltrates in biopsies were unaffected by this H1-antihistamine. Moderate doses of systemic corticosteroids did not alter clinical or histologic reactions at C5a injection sites in two volunteers. This study, using doses within the potential physiologic range of the anaphylatoxin, provides a comprehensive assessment of the effect of human C5a on normal human skin. PMID- 2982915 TI - An in vivo microperfusion study of distal tubule bicarbonate reabsorption in normal and ammonium chloride rats. AB - For many years it has been thought that distal nephron hydrogen ion secretion can be importantly modulated by factors such as sodium delivery, sodium avidity, and potassium stores. Free flow micropuncture studies have also indicated that the rate of bicarbonate delivery may also alter the rate of bicarbonate reabsorption. The present studies were undertaken to examine possible luminal influences on total CO2 reabsorption in microperfused distal tubules in the rat in vivo. Tubules from normal and acidotic rats were perfused with five solutions in a manner that induced changes in bicarbonate load, sodium and potassium fluxes (JNa, JK), and luminal sulfate concentration. in each collected perfusate, simultaneous analyses were undertaken to determine water reabsorption, Na, and K concentrations using graphite furnace atomic absorption spectroscopy and total CO2 by microcalorimetry. Using factorial analysis of covariance to account for confounding effects on total CO2 flux (JtCO2) such as water reabsorption, distal tubules of acidotic rats reabsorbed CO2 in the range of 50-112 pmol X min-1 X mm 1 X These JtCO2 values were not significantly correlated with HCO3 load, JNa, or JK despite changes in the latter from net reabsorption to net secretion. Distal tubules of rats with normal acid-base status had JtCO2 values which were neither significantly different from zero nor correlated with changes in JK and JNa. Further, doubling the load from 250-500 pmol/min (by doubling the perfusion rate of 25-mM HCO3 solutions) did not stimulate JtCO2 in these normal animals. Accordingly, these acute in vivo microperfusion studies indicate for the first time that neither load nor potassium or sodium fluxes are important modulators of distal tubule bicarbonate reabsorption. PMID- 2982916 TI - Comparative study of pituitary-thyroid hormone economy in fasting and hypothyroid rats. AB - Starvation in laboratory rodents results in significant alterations in thyroid hormone economy characterized by decreased circulating levels of thyroxine (T4) and 3,5,3'-triiodothyronine (T3) and a decline in serum thyrotropin (TSH) concentration. To investigate this apparent paradox, we have compared in fasted and hypothyroid animals the intracellular parameters mediating thyroid hormone action in the anterior pituitary gland. In vitro saturation analysis combined with quantitation of nuclear T3 content by radioimmunoassay allowed for characterization of pituitary nuclear T3 receptors and estimation of the endogenous fractional receptor occupancy. In rats, thyroidectomized 4 wk earlier, the 10-fold increase in serum TSH levels and decline in peripheral thyroid hormone concentrations were accompanied by a 61% decrease in pituitary nuclear T3 content and a marked decline in fractional T3 receptor occupancy as compared with control animals. In euthyroid animals subjected to short-term starvation (72 h), serum T3, T4, and TSH levels declined by 52, 43, and 48%, respectively. Despite these marked decreases in circulating thyroid hormone levels, pituitary nuclear T3 content in fasted rats declined by only 15% (P less than 0.05) relative to control levels. This modest decline in nuclear T3 content, combined with a 23% decrease in total T3 receptor number, resulted in an estimated fractional receptor occupancy in fasted animals which was equal to or greater than that noted in controls. The effects of fasting and hypothyroidism on the pituitary were further investigated by quantifying low Michaelis constant (Km) T4 5' deiodinase activity in the crude cytosol fraction of pituitary homogenates. In thyroidectomized animals, maximum velocity was increased ninefold, whereas fasting resulted in a 37% decrease (P less than 0.025) in this parameter compared with controls. Km values were similar in all experimental groups (4.7 +/- 0.6 nM). These results demonstrate that, despite significant reductions in circulating thyroid hormone concentrations and pituitary T4 5'-deiodinase activity, nuclear T3 levels are maintained at relatively normal levels in the pituitary of the fasted animal and fractional T3 receptor occupancy may actually increase. These findings are in marked contrast to those noted in thyroidectomized animals and suggest that the suppression of TSH secretion accompanying starvation in the rat is mediated, at least in part, by local pituitary mechanisms that serve to maintain and possibly enhance nuclear T3 receptor occupancy. PMID- 2982917 TI - Effect of the long-term administration of corticotropin-releasing factor on the pituitary-adrenal and pituitary-gonadal axis in the male rat. AB - The effect of the continuous exposure to ovine corticotropin-releasing factor (oCRF) was measured in adult male rats. The intravenous infusion of 0.75 nmol oCRF/h to intact rats over a 24-h period was accompanied by a peak of ACTH and corticosterone secretion that occurred during the first 90 min of administration of the releasing factor, followed by a decrease to lower, but still above control, values. Additionally, corticotropin-releasing factor (CRF)-treated rats had decreased plasma testosterone levels. The subcutaneous administration of 0.075 or 0.75 nmol oCRF/h to intact rats for 7 d also resulted in elevations of both plasma ACTH and corticosterone levels comparable to those measured after a 24-h exposure to the releasing factor, as well as dose-related hypertrophy of the adrenals and increases in pituitary ACTH content. In these animals, CRF markedly inhibited luteinizing hormone (LH) (but not follicle-stimulating hormone [FSH] ), testosterone, and PRL secretion and decreased seminal vesicle weights. All the effects of CRF were mimicked by exogenously administered ACTH. By contrast, with the exception of FSH secretion, which was slightly elevated by CRF, neither CRF nor ACTH were able to significantly modify reproductive parameters in adrenalectomized animals, which suggests that the elevation of circulating levels of adrenal steroids induced by peripherally administered CRF represents major mediators of CRF-induced inhibition of fertility. These results indicate that in the rat, the continuous stimulation of the pituitary-adrenal axis by peripherally administered CRF causes some degree of desensitization of the pituitary-adrenal axis, but is still accompanied by persistent elevations of the circulating levels of both ACTH and corticosteroids. The increased secretion of adrenal steroids by CRF-treated rats is believed to participate in the disruption of reproductive parameters observed in these rats. PMID- 2982918 TI - Human B lymphoma cell line producing B cell growth factor. AB - Namalva, a human B cell lymphoma line, produced a factor with a molecular weight of approximately 60,000 which enhanced the proliferation of normal activated human B lymphocytes. The factor also enhanced the proliferation of certain B cell lines. It can be distinguished physiologically and biochemically from other lymphokines known to enhance B cell proliferation, namely, interleukin (IL) 1, IL 2, and interferon. The production of B cell growth factor by B cell tumor lines may contribute to their ability to grow autonomously and may reflect an important component of the neoplastic potential of the cell. B cell growth factor produced by tumors may also affect normal cells in vivo. PMID- 2982919 TI - Muscle cell electrical hyperpolarization and reduced exercise hyperkalemia in physically conditioned dogs. AB - Contracting muscle cells release K ions into their surrounding interstitial fluid, and some of these ions, in turn, enter venous plasma. Thereby, intense or exhaustive exercise may result in hyperkalemia and potentially dangerous cardiotoxicity. Training not only reduces hyperkalemia produced by exercise but in addition, highly conditioned, long-distance runners may show resting hypokalemia that is not caused by K deficiency. To examine the factors underlying these changes, dogs were studied before and after 6 wk of training induced by running on the treadmill. Resting serum [K] fell from 4.2 +/- 0.2 to 3.9 +/- 0.3 meq/liter (P less than 0.001), muscle intracellular [K] rose from 139 +/- 7 to 148 +/- 14 meq/liter (P less than 0.001), and directly measured muscle cell membrane potential (Em) in vivo rose from -92 +/- 5 to -103 +/- 5 mV (P less than 0.001). Before training, resting Em of isolated intercostal muscle in vitro was 87 +/- 5 mV, and after incubation in 10(-4) M ouabain, Em fell to -78 +/- 5 mV. After training, resting Em of intercostal muscle rose to -95 +/- 4, but fell to 62 +/- 4 mV during incubation in 10(-4) M ouabain. The measured value for the Em was not completely explained by the increased ratio of intracellular to extracellular [K] or by the potassium diffusion potential. Skeletal muscle sarcolemmal Na,K-ATPase activity (microM inorganic phosphate mg-1 protein h-1) increased from 0.189 +/- 0.028 to 0.500 +/- 0.076 (P less than 0.05) after training, whereas activities of Mg2+ -dependent ATPase and 5'nucleotidase did not change. In untrained dogs, exercise to the point of exhaustion elevated serum [K] from 4.4 +/- 0.5 to 6.0 +/- 1.0 meq/liter (P less than 0.05). In trained dogs, exhaustive exercise was associated with elevation of serum [K] from 3.8 +/- 0.3 to 4.2 +/- 0.4 (NS). The different response of serum [K] to exercise after training was not explainable by blood pH. Basal insulin levels rose from 7.0 +/- 0.7 microU/ml in the untrained dogs to 9.9 +/- 1.0 microU/ml (P less than 0.05) after training. Although insulin might have played a role in the acquired electrical hyperpolarization, the reduced exercise-produced hyperkalemia after training was not reversed by blockade of insulin release with somatostatin. Although the fundamental mechanisms underlying the cellular hyperpolarization were not resolved, our observations suggest that increased Na-K exchange across the sarcolemmal membrane, the increase of Na,K-ATPase activity and possibly increased electrogenicity of the sodium pump may all play a role in the changes induced by training. PMID- 2982921 TI - Simplified restriction endonuclease method for typing and subtyping adenoviruses. AB - Restriction endonuclease digestion of viral DNA labelled in vivo with phosphorus 32 has been used to type and to subtype both conventional and enteric adenoviruses. The method is a modification of that already described for typing and subtyping herpes simplex virus and, like the latter, needs far less material than methods previously described. PMID- 2982923 TI - View from the nation's capital. PMID- 2982922 TI - Incidence of anti-intermediate filament antibody in serum samples of students with suspected glandular fever. AB - Serum samples from 40 students with suspected infectious mononucleosis were tested for the presence of antibodies to intermediate filaments (AIFA) of the cytoskeleton. Twenty had antibodies to the Epstein-Barr virus capsid antigen before their illness, and during it their sera remained negative by the Paul Bunnell test. The other 20 patients did not have antibodies to the Epstein-Barr virus capsid antigen before their illness and seroconverted during the illness. These patients (true infectious mononucleosis group) developed positive Paul Bunnell tests. Sera from normal subjects (blood donors) were also tested for AIFA. AIFA was present in titres greater than 1/10 in 80% of the infectious mononucleosis group (mean titre 1/40-1/80), 10% of the Paul-Bunnell negative glandular fever group, and 8.5% of the normal blood donors. PMID- 2982920 TI - The pathobiology of the osteoclast. AB - This article reviews recent information concerning the origin of osteoclasts and the local and systemic regulation of their activity. It appears that much of the environmental responsiveness of osteoclasts is mediated by cells of the osteoblastic lineage, which exert a major influence on the localisation, induction, stimulation, and inhibition of osteoclastic bone resorption. Some of the mechanisms by which osteoclast function may be disturbed by inflammatory and neoplastic diseases are discussed, and it is suggested that many pathological disturbances of osteoclastic bone resorption may be explicable as mimicry of physiological regulatory mechanisms by local hormones introduced into bone as the local regulators of the diseased tissue. PMID- 2982924 TI - Ultrastructure and synaptic contacts of enkephalinergic amacrine cells in the retina of turtle (Pseudemys scripta). AB - Bistratified amacrine cells of the turtle retina containing enkephalin-like immunoreactivity were examined with the electron microscope with the aid of peroxidase immunocytochemical techniques. Our goal was to determine the nature and the location of the synaptic contacts of these cells and the intracellular localization of the immunoreactivity. There was a diffuse reaction product throughout the cytoplasm which coated the surfaces of all the organelles and a dense reaction product which filled the core of some large cytoplasmic vesicles (130 nm in dia.). These labeled amacrine cells received conventional synaptic contacts from other unlabeled amacrine cells and ribbon synaptic contacts from unlabeled bipolar cells, in both the proximal and distal inner plexiform layer. These enkephalin-positive amacrine cells made conventional synaptic contacts containing unlabeled synaptic vesicles (60 nm in dia.), with ganglion cells in the proximal inner plexiform layer and with bipolar cells in the distal inner plexiform layer. These results suggest that enkephalin-like material coexists with another neurotransmitter within these neurons and that these amacrine cells are able to integrate information from both amacrine cells and bipolar cells and provide synaptic input to bipolar cells, ganglion cells, and possibly other amacrine cells. PMID- 2982925 TI - Morphology of identified interneurons in the CA1 regions of guinea pig hippocampus. AB - Identified interneurons in the CA1 region of guinea pig hippocampus were examined using light and electron microscopic (EM) techniques. The HRP was intracellularly injected into cells, recorded in the in vitro slice preparation, which met physiological criteria for interneurons. These neurons were characterized at the light and electron microscopic level, and used as standards for identifying interneurons which had not been HRP-labeled. Pyramidal basket cell somata were found in stratum oriens and stratum pyramidale; they were easily identifiable by their convoluted nucleus, dense endoplasmic reticulum, and numerous organelles. Aspinous dendrites reached into stratum oriens, where they received profuse synaptic input (primarily asymmetric synapses). Many of the asymmetric synapses degenerated following commissural lesions, suggesting that much of the input to interneurons was from extrinsic afferents. Dendrites were characterized by their spindled appearance, especially at distal sites. They showed postsynaptic degenerative changes following commissural lesions. Interneuron axons were extremely fine, with regular enlargements or "beads" which made apparent synaptic contacts primarily on pyramidal cell somata. The axon of a single, HRP-injected interneuron made many apparent contacts on large numbers of pyramidal cells; axons arborized over distances of several hundred micra within stratum pyramidale. This study provides direct evidence that neurons, with an identified inhibitory interneuron function in hippocampus, can mediate feed-forward as well as feed-back (recurrent) inhibition. Interneuron output showed extreme divergence, with influence over large distances. The high density of intracellular organelles in these cells suggested high metabolic activity and demand, perhaps making these interneurons exceptionally vulnerable to trauma induced damage. PMID- 2982926 TI - (3H) glycine-accumulating neurons of the human retina. AB - Isolated human retinas were incubated in physiological saline containing micromolar (3H) glycine. The types, distributions, and synaptologies of glycine accumulating neurons were determined by light and electron microscope autoradiography. Two types of amacrine cells were discriminated on the bases of number of processes descending into the inner plexiform layer, density of label in light-microscope autoradiographs, size, and synaptic features: (1) Gly1 amacrine cells have moderate labeling, several oblique dendrites arising from the soma, and electron lucent synaptic terminals containing large presynaptic specializations, nd (2) Gly2 amacrine cells have dense labeling, a single proximal dendrite, and moderately electron-dense terminals with small presynaptic specializations. Gly1 amacrine cells constitute approximately 15% and Gly2 amacrine cells approximately 38% of all cells in the amacrine cell layer. The laminar distribution of label in the inner plexiform layer was measured by scanning microdensitometry, which provided a format for categorizing types of synaptic contacts. Many features of glycine-accumulating amacrine cell contacts were similar to those of cat AII/Gly2 amacrine cells: a diffuse yet bisublaminar distribution of label, concentration of synaptic output in sublamina a, rod bipolar cell input in sublamina b and gap junctions in mid-inner plexiform layer involving labeled cells. The evidence seems to indicate that human Gly2 amacrine cells and cat AII/Gly2 amacrine cells are homologous cell types. finally, some cone bipolar cells were labeled. PMID- 2982927 TI - Observation of Pigeon herpesvirus 1 re-excretion during the reproduction period in conventionally reared homing pigeons. AB - Pigeon herpesvirus 1 (PHV1) re-excretion by latently infected mature homing pigeons was followed during the reproduction period under natural conditions. All latently infected pigeons re-excreted PHV1 during the reproduction period and most of them during squab-gorging. PMID- 2982928 TI - A malignant liver cell tumour in a king snake (Lampropeltis getulus). AB - A neoplastic lesion in the liver of a king snake, Lampropeltis getulus, was identified as a hepatocellular carcinoma. There were metastases in the spleen. PMID- 2982929 TI - Metamorphosis of dermal cylindroma: possible relation to malignant transformation. Case report of cutaneous cylindroma with direct intracranial invasion. AB - A case of locally invasive cutaneous cylindroma is reported. This neglected scalp tumor, 6.5 cm in greatest dimension, initially invaded the calvaria at the time of the first operation. A recurrence infiltrated the dura and presented as an intracranial space-occupying lesion. No brain invasion was present and distant metastases were not documented. The patient remains well 2 1/2 years after tumor resection. Light and electron microscopic appearance of the recurrent tumor revealed distinct morphologic features, which may be of use in predicting the biologic behavior of cutaneous cylindromas. This case documents the low biologic aggressiveness of cylindroma and stresses the necessity of complete surgical excision of neoplasms with atypical morphologic features. PMID- 2982930 TI - Use of Burow's wedge principle for repair of wounds in or near the eyebrow. AB - Burow's wedge flap provides an excellent method for repair of wounds in or near the eyebrow. Advantages afforded by this technic in this context include ample flap elasticity and vascularization, versatility in dealing with wounds of varying dimension and location, and fewer incisions in comparison with the number necessary with other methods. Between 1980 and 1984 the author used this method in twelve patients and obtained good to excellent results in eleven and a fair outcome in one. PMID- 2982932 TI - MR imaging of portal vein thrombus in hepatocellular carcinoma. AB - This report describes the magnetic resonance (MR) demonstration of portal vein tumor thrombus (PVTT) in hepatocellular carcinoma (HCC). In two cases of HCC with PVTT diagnosed by angiography, MR demonstrated PVTT as areas of increased MR signal in the portal vein. Our limited experience suggests the potential utility of MR imaging for diagnosis of PVTT. PMID- 2982931 TI - Imaging opiate receptors in the human brain by positron tomography. AB - Carfentanil is a potent, synthetic opiate that binds to mu opiate receptors with very high affinity (KI = 0.051 nM, 37 degrees C). In rat brain, carfentanil is 90 and 250 times more selective for mu opiate receptors compared with delta and kappa opiate receptors, respectively. Carbon-11-carfentanil was synthesized by reacting [11C]methyliodide with the carboxylic acid precursor of carfentanil. Carbon-11-carfentanil was injected intravenously in man and a baboon and the distribution of the radioactivity in the brain was monitored using positron emission tomography. High concentrations of radioactivity were observed in the basal ganglia and thalamus, intermediate concentrations in the frontal and parietal cerebral cortex, and low concentrations in the cerebellum and occipital cortex; this distribution corresponds to the known regional density of opiate receptors measured using in vitro techniques. This heterogeneous distribution could be abolished by pretreatment with naloxone (1 mg/kg), an opiate antagonist. The percent inhibition of binding by naloxone is approximately 90% in the caudate nucleus and medial thalamus for the period 30-60 min after injection; therefore, this method is associated with a high level of specific binding to opiate receptors compared with nonspecific binding sites. The ability to measure opiate receptors in vivo in man makes it possible to study a variety of neurologic and psychiatric disorders in which opiate receptors are thought to be abnormal and to study physiologic role of opiate receptors in the central nervous system. PMID- 2982933 TI - Syringadenocarcinoma papilliferum. AB - A giant tumor developed on the right chest wall of a 52-year-old housewife over a 20-year period. It was accompanied by a spider-like invasion of skin, and a lymph node was palpated in the right axilla. Wide excision of the lesion was carried out. The cross-section of the tumor showed two different appearances. The tumor was histologically diagnosed as a syringadenocarcinoma papilliferum. The post operative course was uncomplicated and without evidence of recurrence. PMID- 2982934 TI - Dermal cylindromas (turban tumour) and eccrine spiradenomas in a patient with membranous basal cell adenoma of the parotid gland. AB - A patient with coexisting dermal cylindromas (Turban tumour), eccrine spiradenomas and a variant of basal cell adenoma of the parotid gland characterized by prominent focal deposition of hyaline (membranous basal cell adenoma of parotid gland) is reported. The microscopic appearance of the cutaneous and parotid gland tumours was very similar. This rare association of analogous tumours of sweat and parotid glands has been described only in 5 previous cases. It is hypothesized that the occurrence of the 3 different tumours found in this individual may be explained by the multipotential reserve cell concept and these tumours perhaps could be considered as different expressions of a single genetic disorder. PMID- 2982935 TI - Properties of microfilled composite resins as influenced by filler content. AB - Two series of composite resins were prepared with a light-cured urethane dimethacrylate matrix to which varying amounts of two types of silanated silica particles were added. One series contained volume fractions ranging from 15.8 to 28.8% silica particles of 20 nm in diameter (Type I filler) and the other series volume fractions of from 24 to 49.4% of an agglomerated silica particle of 40 nm in diameter (Type II filler). Tests were conducted to determine the effect of filler level on: depth of cure as determined by hardness measurements; color stability in both UV light and water; water sorption with time; hardness; compressive strength; strain behavior in slow compression; and resistance to toothbrush abrasion and wear by hydroxyapatite. Analysis of the data obtained for these two microfilled series indicate that increased filler levels result in trends for increased depth of cure, color stability, hardness, compressive strength, and stiffness, while water sorption and resistance to both toothbrush abrasion and wear by hydroxyapatite were reduced. These trends were more pronounced for the Type II filler series than for the Type I filler series. However, there was a greater differential in filler levels within the Type II series than within the Type I series. PMID- 2982936 TI - Contribution of stereochemical interactions in the adhesion of Streptococcus sanguis C5 to experimental pellicles. AB - Parameters for the adsorption of Streptococcus sanguis C5 to experimental salivary pellicles were determined to assess the relative contributions of stereochemical and less specific hydrophobic-electrostatic interactions in this process. S. sanguis C5 cells possess hydrophobic surfaces and also an adhesin which exhibits stereochemical binding to sialic acid-containing salivary receptors. The adsorption of S. sanguis C5 to untreated pellicles was significantly better described by a two-site adsorption model than by a one-site model (p less than 0.001). One class of binding sites had an affinity for the organism that was more than 400 times that of the second class of binding sites. However, adsorption of the organism was better described by a one-site model to asialo pellicles prepared by neuraminidase treatment which destroyed the receptors for the organism's adhesin. The affinity of the organism to sites in asialo pellicles was low, and approximated that to the low-affinity sites in untreated pellicles. These observations suggest that, in the absence of stereochemical interactions involving the organism's adhesin and specific salivary receptors, S. sanguis C5 cells only exhibit low-affinity binding to experimental pellicles. This was substantiated by studies which showed that adsorption of the organism to albumin-coated hydroxyapatite and to polystyrene beads was also better described by a one-site model, and it occurred with only low affinity. PMID- 2982937 TI - Cyclic adenosine 3', 5'-monophosphate levels in human parotid saliva at different secretion rates. AB - Seventy-four samples of unstimulated and stimulated parotid saliva were collected from 12 healthy male volunteers. The secretion rates and cAMP concentrations were determined. The secretory rate ranged from 0.01 to 1.03 ml/min/gland. The concentration of cyclic AMP varied from 1.6 to 31.9 pmol/ml. The concentration of cAMP was 10.0 +/- 6.5 pmol/ml in unstimulated saliva and 3.8 +/- 1.6 pmol/ml in stimulated saliva. A curvi-linear relationship existed between secretory rate and cyclic AMP concentrations, and the best-fitting regression equation was y = 2.06 + 0.5/x, where y = concentration of cyclic AMP and x = secretory rate. PMID- 2982938 TI - Evaluation of dispersible formulations of erythrosin B for field control of Culex pipiens quinquefasciatus Say (Diptera: Culicidae). PMID- 2982939 TI - The joint action of coprecipitated free-acid formulations of erythrosin B and fluorescein against the larvae of Culex pipiens quinquefasciatus Say (Diptera: Culicidae). PMID- 2982940 TI - Cyst and cystlike lesions of the foot. AB - A variety of cyst and cystlike lesions may present in the osseous structures of the foot. Such lesions are infrequent in their occurrence, and often cannot be diagnosed on the basis of radiographic appearance. In addition to the morphologic data that can be derived from evaluation of standard radiographs, other diagnostic radiographic studies such as tomography, CT scanning, radionuclide bone imaging, angiography, or other studies may be required to ascertain the nature and extent of cystlike lesions of the foot. This is dependent on histopathologic information derived from biopsy. A variety of cyst and cystlike lesions of the foot are presented with a review of their basic morphology and histopathologic, clinical, and prognostic characteristics. PMID- 2982941 TI - Biochemical techniques for the genetic and phenotypic analysis of viruses: 'molecular epidemiology'. PMID- 2982943 TI - Coexpression of T4 and T8 on peripheral blood T cells demonstrated by two-color fluorescence flow cytometry. AB - Using two-color fluorescence flow cytometry, we were able to detect the presence of small numbers of T4+T8+ cells (about 3%) in freshly isolated peripheral T cell populations derived from normal healthy donors. Coexpression of T4 and T8 was predominantly found on large blastlike cells and appeared to be related to activation. Stimulation of peripheral T cells with concanavalin A (Con A) for 5 days resulted in the generation of up to 60% of T4+T8+ cells. Coexpression was accompanied by a twofold increase in the number of T8 antigenic sites per cell. The T4+T8+ cells in lectin-stimulated cultures expressed high levels of the activation antigens T9, T10, and the IL-2 receptor but lacked T6, an antigen found on a majority of stage II thymocytes. Coexpression of T4 and T8 appeared to be a transitory process, because prolonged culture of T cells in the absence of lectin resulted in the loss of the T4+T8+ phenotype. Our data suggest that T cell activation in peripheral blood results in the generation of a T4+T8+ cell population which is distinct from previously described thymic and peripheral blood cells. Because T4 and T8 molecules may interact directly with MHC antigens, coexpression of these molecules may have an important role in immune function. PMID- 2982942 TI - Immunofluorescent staining of trypsinized formalin-fixed brain smears for rabies antigen: results compared with those obtained by standard methods for 221 suspect animal cases in Nigeria. AB - Formalin-fixed samples from 221 animal brains received for rabies diagnosis in Nigeria were digested in 0.1% trypsin in phosphate buffered saline, pH 7.4, and smears stained for rabies antigen by direct immunofluorescence (IF). The results were compared with those obtained using fresh material from the same animals for Negri body staining, mouse inoculation (MI) and occasionally immunofluorescent staining. From 191 specimens examined for Negri bodies and by mouse inoculation 51 and 64 respectively proved positive. The IF smear technique under investigation failed to detect 5 of these but showed up as positive 30 which had been recorded as Negri-negative and 19 that had gone undetected by MI too. In a direct comparison with IF staining of fresh tissue from 23 known rabies positive animals the similar staining of trypsin-digested formalized smears failed to give a positive result in 2 out of 23 cases. Some problems were encountered with smears not sticking to slides. When gelatinized slides were used fluorescence was sometimes poorer. Where transport and refrigeration are difficult and section cutting equipment is lacking the technique of IF staining of smears prepared from formalized brain tissue after treatment with trypsin can be a useful adjunct to other diagnostic methods. It also makes for safer working where special facilities are absent. PMID- 2982944 TI - Human splenic sinusoidal lining cells express antigens associated with monocytes, macrophages, endothelial cells, and T lymphocytes. AB - The antigenic and functional properties of splenic sinusoidal lining cells (SLC) have not been studied extensively. Some investigators have suggested that SLC are actively phagocytic, and thus part of the mononuclear phagocyte system. Others dispute this and assign the functions of endothelial cells to the SLC. During studies in situ of phenotypic subpopulations of human splenic macrophages (M phi), we found that SLC share membrane antigens (HLA-DR and OKM5), an enzyme (lysozyme), and histochemical properties (nonspecific esterases) with monocytes and M phi. In addition, we showed that LSC, like endothelial cells, synthesize factor VIII of the clotting system and also bear the receptor for transferrin. Our previous studies found that SLC express the antigens found on helper/inducer (OKT4, Leu-3a,b) and suppressor/cytotoxic (OKT8, Leu-2a) T lymphocyte subsets. We have confirmed these observations, and have shown by means of preincubation with soluble complexes of anti-human IgG-human IgG that the detection of T cell and other antigens on SLC is not due to nonspecific binding of antibodies by Fc receptors. By using techniques designed to isolate and purify splenic M phi, we were able to obtain SLC in suspension and to demonstrate that they retain the antigens detected in situ. Thus, the human splenic SLC expresses a unique combination of antigens, histochemical properties, and cell products in common with monocytes, M phi, and T lymphocytes. PMID- 2982945 TI - Activated human B cells display a functional IL 2 receptor. AB - These studies deal with the expression of a functional IL 2 receptor on activated primary human B cells. Antibody against the receptor (alpha-TAC) reacted with 25 to 65% activated B cells, inhibited B cell proliferation by 50% and inhibited B cell secretion of Ig by greater than 90%. These effects were shown to be independent of contaminating T lymphocytes. Anti-TAC immunoprecipitated a molecule of identical size (65,000 daltons) from T and B lymphocytes; B cells were also shown to actively synthesize the IL 2 receptor. The chymotryptic peptide chromatograms of TAC antigen from T and B cells show these molecules to be indistinguishable. PMID- 2982946 TI - Human B cell responsiveness to B cell growth factor after activation by phorbol ester and monoclonal anti-mu antibody. AB - The effect of phorbol ester on human B cell activation was examined. Picomolar to nanomolar concentrations of phorbol ester induced a high level of proliferation in small IgM-positive B cells isolated from peripheral blood by fluorescence activated cell sorting. The addition of optimal doses of anti-mu antibody resulted in enhanced proliferation of phorbol ester-activated B cells. The addition of B cell growth factor (BCGF) to phorbol ester-activated B cells also resulted in a dose-dependent synergistic effect and maximal enhancement on day 3. BCGF activity could be absorbed with either phorbol ester- or anti-mu-activated B cells, but not with resting B cells, thus confirming the induction of functional BCGF receptor expression. Cell proliferation was not necessary for the induction of functional BCGF receptors. Phorbol ester was a more efficient inducer of BCGF receptor expression than was anti-mu antibody; gamma-interferon treatment had no effect. BCGF enhanced transferrin receptor expression by phorbol ester-activated B cells. The results suggest that phorbol ester-activated small B cells can be used to monitor BCGF activity, and this synergistic combination may be useful in establishing BCGF-dependent B cell clones in culture. PMID- 2982947 TI - Intracellular retention of the 5-lipoxygenase pathway product, leukotriene B4, by human neutrophils activated with unopsonized zymosan. AB - The cellular and extracellular distribution of leukotriene B4 (LTB4) generated in human neutrophilic polymorphonuclear leukocytes (PMN) stimulated with unopsonized zymosan has been compared with that generated in PMN activated by the calcium ionophore. The amounts of extracellular and intracellular LTB4 were quantitated by radioimmunoassay. The authenticity of the immunoreactive LTB4 was confirmed by the elution of a single immunoreactive peak after reverse phase-high performance liquid chromatography (RP-HPLC) at the retention time of synthetic LTB4, by the identical elution time of a peak of radiolabeled product derived from [3H]arachidonic acid-labeled PMN with the immunoreactive product, and by the comparable chemotactic activity on a weight basis of immunoreactive LTB4 and synthetic LTB4 standard. Under optimal conditions of stimulation by unopsonized zymosan, more than 78% of the generated immunoreactive LTB4 remained intracellular, whereas with optimal activation by the ionophore, less than 8.6% of immunoreactive LTB4 was retained. Resolution by RP-HPLC of the products from the supernatants and cell extracts of [3H]arachidonic acid-labeled PMN stimulated with unopsonized zymosan and those stimulated with calcium ionophore allowed identification and measurement of 5-hydroxyeicosatetraenoic acid (5-HETE), 6 trans-LTB4, LTB4, and omega oxidation products of LTB4 by radioactivity. With zymosan stimulation of PMN, 5-HETE and the 6-trans-LTB4 diastereoisomers were not released, LTB4 was partially released, and the omega oxidation products of LTB4 were preferentially extracellular in distribution. In contrast, with ionophore stimulation, only 5-HETE had any duration of intracellular residence being equally distributed intra- and extracellularly throughout the 30-min period of observation; 6-trans-LTB4, LTB4, and the omega oxidation products of LTB4 were retained at less than 19%. The respective distributions of 5-HETE after zymosan and ionophore stimulation were not altered by the introduction of albumin to the reaction mixtures to prevent reacylation, or by hydrolysis of the cell extract to uncover any product that had been reacylated. The finding that stimulation of PMN with unopsonized zymosan results in the cellular retention of 5-lipoxygenase products suggests that release of these metabolites may be an event that is regulated separately from their generation. PMID- 2982948 TI - Natural killer cell-mediated lysis involves an hydroxyl radical-dependent step. AB - The role of oxygen radicals in lysis of K562 target cells by human natural killer (NK) cells was determined by addition of scavengers of these free radicals. Lysis was greatly reduced under hypoxic conditions. Superoxide dismutase and cytochrome c, scavengers of superoxide anions, and catalase and scavengers of hypochlorite had no effect on lysis. Of 15 hydroxyl radical scavengers tested, 13 inhibited lysis. These were not toxic, because cell morphology and spontaneous chromium release were not affected and preculture with scavengers was not inhibitory. These scavengers differed widely in structure, but degree of inhibition of lysis correlated with their rate constants (k) for reaction with hydroxyl radical (k vs log inhibitor concentration required to decrease lysis by 50%: r = -0.9202, p less than 0.001), showing that inhibition was due to inactivation of the hydroxyl radical. Target cell binding was not reduced at concentrations that inhibited lysis. Inhibitors of the lipoxygenase pathway also decreased lysis, suggesting this pathway to be the source of hydroxyl radicals. In view of the reported requirements for hydroxyl radical-mediated lipid peroxidation for optimal secretory activity in a number of cell types, it appears that the generation of hydroxyl radicals by NK cells is required for delivery of cytotoxic factors. PMID- 2982949 TI - Human natural killer cells limit replication of herpes simplex virus type 1 in vitro. AB - Studies were undertaken to determine whether natural killer (NK) cells could inhibit the replication of herpes simplex virus type 1 (HSV-1) in culture. In the absence of effector cells, HSV-1 was found to replicate in fibroblasts with up to a 100-fold increase in virus titer from 4 to 16 hr after incubation at 37 degrees C. Human peripheral blood mononuclear cells were found to limit virus replication in a dose-dependent manner, with the greatest inhibition being observed at the highest concentration evaluated: i.e., an effector:target ratio of 800:1. The antiviral effect was not observed when nonactivated or virus-activated mononuclear cells were added to the virus preparations at the end (instead of the beginning) of the assay period, indicating that the observed effect was not due to a nonspecific toxicity of soluble factors released from freeze-thawed effectors. Neither was inhibition of HSV-1 replication due to the generation of interferon (IFN) during the NK assay, because the addition of anti-IFN did not abrogate the antiviral effect. Thus, the inhibition of viral replication was most likely due to a cytotoxic effector rather than to release of soluble factors. The effector cells responsible for limiting HSV-1 replication were shown to be NK cells by a number of criteria. Mononuclear cells from both HSV-1 seropositive and seronegative donors limited virus replication; their activity could be boosted by pretreatment of effector cells with IFN; the effector cells which limited virus replication were found in Percoll gradient fractions enriched for large granular lymphocytes; and the effector cells shared the cell surface phenotype of NK cells -they were enriched in populations depleted of T cells by panning with Leu-4 and were depleted of activity by treatment with the anti-NK antibody Leu-11b plus complement. We conclude that human NK cells are capable of recognizing and lysing HSV-1-infected target cells before infectious virus progeny are generated. These results suggest that NK cells, acting early in the course of an infection, might serve to limit HSV-1 replication and therefore reduce the virus load in the host before the development of the adaptive immune response and clearance of the infection. PMID- 2982950 TI - Herpes simplex virus type 1 infection of endothelial, epithelial, and fibroblast cells induces a receptor for C3b. AB - We recently demonstrated that herpes simplex virus type 1 (HSV 1) induces a receptor on human umbilical vein endothelial cells for complement component C3b (C3bR). We assigned this receptor function to HSV 1 viral glycoprotein C (gC) based on several observations: tunicamycin, which prevents glycosylation and expression of N-linked glycoproteins on the surface of infected cells, markedly reduced expression of the C3bR; monoclonal antibodies to HSV 1 gC blocked detection of the C3bR, whereas monoclonal antibodies to other HSV 1 glycoproteins (gB, gD, gE) had no effect; and the MP mutant of HSV 1, which fails to express gC, did not induce C3bR. We now report that HSV 1 induces C3bR on a wide variety of cell types including bovine thoracic aorta and pulmonary artery endothelial cells, human embryonic lung and embryonic foreskin fibroblasts, and human embryonic kidney cells. To date, all cells studied that are permissive to HSV 1 express C3bR, although the pattern of rosetting of C3b-coated erythrocytes varies among the cell strains examined. We also demonstrate that C3bR expression is not a general response of human umbilical vein endothelial cells to injury, because three other viruses (adenovirus 7, measles, and mumps) do not induce C3bR after infection of these cells. Previously we had shown that among herpes simplex viruses, a variety of HSV 1 strains induce C3bR, whereas HSV 2 strains do not. We now demonstrate that other herpes family viruses (CMV and VZV) do not express C3bR. Therefore, C3bR expression appears to be unique for HSV 1 and occurs on a wide variety of cells permissive to this virus. PMID- 2982951 TI - Cloning and complete sequence of an HLA-A2 gene: analysis of two HLA-A alleles at the nucleotide level. AB - The gene for the HLA-A2 antigen has been cloned from the human lymphoblastoid cell line 721. Comparison of this sequence with the published sequence for HLA-A3 permits the examination of two alleles at the extremely polymorphic HLA-A locus. A high degree of sequence conservation was seen in both coding and noncoding DNA, 97.2% and 94.5%, respectively. Interestingly, the 3' untranslated region was the most conserved, with 99.5% homology. The polymorphism of the HLA-A antigens results from a high proportion of amino acid substitutions relative to the total nucleotide changes in exons 2 and 3. Unlike the clustered differences seen in this region on comparison of two H-2K alleles of mouse, nucleotide substitutions between the HLA-A2 and A3 alleles are evenly distributed. Substitutions at silent sites and within introns were used to calculate an intra-allelic divergence time of at least 10 to 15 million years for these two HLA-A alleles. PMID- 2982952 TI - The Igh-V locus of MRL mice: restriction fragment length polymorphism in eleven strains of mice as determined with VH and D gene probes. AB - The MRL strain of mice is a model system that closely parallels the human autoimmune disease systemic lupus erythematosus. Our analysis of the variable region genes of MRL mice showed that the MRL/lpr D genes were similar to those of the C3H mouse (Igh-C allotype j). This result was unexpected, because previous studies of the MRL/lpr and MRL/+ substrains suggested that they are allotype a at the Igh-1 (gamma 2a) locus of the constant region. The Igh-V (heavy chain variable region) locus of the MRL/lpr and MRL/+ strains of mice and their parents were therefore examined by restriction fragment length polymorphism with probes for the DSP2 and DFL16 gene families and with two cloned VH probes. Five other strains of mice were also included because the heavy chain locus of the LG mouse, which is the major progenitor of the MRL strains, has not been studied. The MRL substrains and the LG and C3H parents were indistinguishable at all the Igh-V loci studied. These results suggest that the MRL substrains and their LG parent are haplotype j at the Igh-V locus. The results obtained with D gene probes show that the DSP2 gene family is more polymorphic than the DFL16 gene family, which is relatively conserved. We have assigned Igh-V haplotypes for the four VH loci to the 11 strains of mice studied. PMID- 2982954 TI - Dimethyl sulfoxide induces expression of H-2 antigens on mouse lung carcinoma cells. AB - The addition of dimethyl sulfoxide (DMSO) to cultures of line 1 carcinoma cells can increase the surface expression of H-2K and H-2D antigens at least 100-fold from barely detectable initial levels, as determined by using specific monoclonal antibodies and flow cytometry. H-2 values stabilize approximately 1 wk after exposure to maximally inducing concentrations of DMSO (3% vol) at densities found on normal spleen cells. Increased expression of H-2 antigens is not the result of cell selection, it is specific in that expression of an unrelated surface protein decreases, and it is associated with increased synthesis of these antigens as measured by incorporation of [35S]methionine. Additional DMSO-induced changes in the growth, cycling, lectin binding, and antigenic properties of line 1 cells are consistent with increased cell maturation. All changes are reversed when DMSO is removed. This system may facilitate study of products associated with differentiation that influence tumor cell malignancy. PMID- 2982953 TI - Combinational diversity within variable regions bearing the predominant anti-p azophenylarsonate idiotype of strain A mice. AB - The humoral immune response in strain A mice to protein conjugates of p azophenylarsonate (Ars) is characterized by the presence of a major cross reactive idiotype denoted as IdCR. Previous molecular analyses of monoclonal IdCR+ Ars-binding antibodies isolated from multiply immunized animals have indicated that these antibody variable (V) regions may be the expressed product of a single combination of VH, D, JH, V kappa, and J kappa gene segments. The basis of this apparent domination of the Ars response by V regions encoded by this single combination of gene segments is unclear, but is discussed in this report. Our structural analyses on five monoclonal IdCR+ antibodies that are unable to bind Ars show that in contrast to those of Ars-binding IdCR+ antibodies, these (Ars-nonbinding) IdCR+ V regions are encoded by multiple combinations of VH, D, JH, V kappa, and J kappa gene segments, but with the commonality that they all utilize a single VH gene segment (VHIdCR). We provide examples in which the VHIdCR gene segment is expressed with three different V kappa gene segments and with each of the four JH gene segments to produce serologically detectable IdCR+ Ars-nonbinding antibodies. It would thus appear that the previous failure to detect alternative IdCR+ V segment combinations was due to a sampling procedure requiring that the IdCR+ antibody bind Ars, and not the result of restricted assembly or expression of the VHIdCR gene segment with a particular combination of D, JH, V kappa, and J kappa gene segments. This bias in protocol, however, cannot completely account for the homogeneity in previously studied IdCR+ Ars-binding antibodies, because we were able to isolate, from primary immune responses, IdCR+ antibodies that do bind Ars but that utilize alternative V segment combinations. This finding suggests that combinations of V gene segments encoding IdCR+ antibodies are more numerous in primary as opposed to secondary immune responses, and raises the question of why a single combination of VH, D, JH, V kappa, and J kappa gene segments dominates the secondary strain A immune response to Ars. PMID- 2982955 TI - Fixed herpes simplex type 1-infected cells as antigen for in vitro lymphocyte proliferation. AB - Fixed herpes simplex virus type 1 (HSV-1)-infected Vero cells were used as antigen in the in vitro lymphocyte reactivity (LR) test and compared with extracellular HSV, HSV-infected cell extract and purified virions. The highest LR was measured after an incubation period of lymphocytes with the fixed HSV infected Vero cells of 5-7 days. The LR appeared to be dependent on the lymphocyte to fixed HSV-infected cell ratio and was found to be optimal at a ratio of 10-20. The fixed HSV-infected cells could be stored at 6 degrees C without detectable loss of LR. Addition of high-titered anti-HSV pooled serum to the lymphocyte cultures with the fixed HSV-infected cells as antigen inhibited the LR. The highest reactivity was found using HSV-negative pooled serum. Lymphocytes from seropositive donors were stimulated by the fixed HSV-infected cells and the purified virions. LR to extracellular HSV and an extract of HSV infected cells were negative for 5 and 2 out of 13 seropositive donors, respectively. Lymphocytes from seronegative donors were not stimulated by any of the HSV-antigen preparations. Fixed HSV-infected cells, which have the advantage that they are easy to prepare and can be stored at 6 degrees C for several months, are a good alternative to purified HSV-1 virions in the LR test. PMID- 2982956 TI - What does epidermal growth factor do and how does it do it? PMID- 2982957 TI - Microbial resistance to newer generation beta-lactam antibiotics: clinical and laboratory implications. AB - Certain nonfastidious, gram-negative bacilli possess the ability to rapidly develop resistance to many of the newer "enzyme stable" beta-lactam antibiotics. This finding poses many clinical problems including emergence of resistance during therapy with the drugs. Therapeutic alternatives for patients are severely limited when this problem occurs because multiple drug resistance may arise simultaneously. To date, two mechanisms have been found to be responsible for this resistance. The first, which produces multiple beta-lactam resistance, is the induction of chromosomal beta-lactamases that mediate resistance to nonsubstrate drugs by the creation of a nonhydrolytic barrier that blocks access to target proteins within the cell. The second mechanism, which produces beta lactam/aminoglycoside resistance, involves a change in outer membrane permeability. Outbreaks of nosocomial infections with these multiple drug resistant organisms and spread of the strains throughout the hospital are already being seen. Control of these problems can only be achieved through the judicious and restricted use of these new antibiotics. PMID- 2982958 TI - Newer directions in vaccine development and utilization. AB - The gradually evolving technology for vaccine development from Jenner to recombinant genetics has provided both solid accomplishment and possible bases for prophylactic control of essentially all the infectious diseases of humans. The present review gives a prospective view of future vaccines and the new biotechnology as illustrated mainly in the examples of vaccines for control of hepatitis and of herpesvirus infections. Although vaccines offer great benefit for human health, their use is restricted in developing countries by lack of funds and in developed countries such as the United States by failure of application, mainly in adults. Practical matters relating to vaccine use must be resolved if there is to be justification for vaccine development. PMID- 2982959 TI - Oral poliovirus vaccine: history of its development and use and current challenge to eliminate poliomyelitis from the world. AB - Oral poliovirus vaccine (OPV) is like no other live virus vaccine used in humans: vaccine strains multiply extensively in the intestinal tract, are widely disseminated in the family and community, and immunize a large proportion of the unvaccinated population. During the search for optimal strains for vaccine use, motor neurons in the spinal cord of chimpanzees (and by extrapolation those of humans) were found to be much more resistant to polioviruses than those of monkeys; the reverse was true for the alimentary tract. Various biologic properties of polioviruses also varied quantitatively over a wide spectrum and were genetically distinct. The phenomenon of somewhat increased neurovirulence for monkeys, but not for chimpanzees, encountered in excreted virus was extensively studied in families, in children's homes, and finally among hundreds of thousands of susceptible children and adults in areas where only 50% of the susceptible population received OPV; these studies did not reveal evidence of danger. During the past 20 years approximately 5 million cases of paralytic poliomyelitis were probably prevented by OPV in predominantly temperate-climate countries inhabited by approximately 2 billion people. OPV has also been used less extensively and not optimally in many tropical and subtropical countries, where paralytic poliomyelitis is now known to be an important public health problem, with reduction in numbers of cases but not elimination of the disease except in some countries with better health services. Experience in Cuba during the past 21 years, in Brazil during the past 5 years, and in the Dominican Republic during the past 2 years has shown that the strategy of annual short-term vaccination of all children in the most susceptible age groups can rapidly eliminate the disease from tropical and subtropical countries. PMID- 2982960 TI - Adenoviruses and pediatric gastroenteritis. AB - Adenoviruses were found in 8.6% of 900 pediatric inpatients with diarrhea who were tested by electron microscopy of a fecal specimen and cell culture inoculation of a throat swab and an anal swab specimen. In 5.1% of these patients, including 13.5% of patients who were four through five months of age, adenovirus particles were visualized in the fecal specimen. Controlled study demonstrated that visualized adenoviruses, especially those that did not grow readily in conventional Hep-2 cell cultures, were significantly associated with diarrhea. About 80% of the visualized adenoviruses from patients with diarrhea or vomiting or both, including 94% of the viruses that grew in the 293 cell line but that did not grow readily in Hep-2 cultures, proved to be enteral adenoviruses- adenoviruses from either group F (type 40) or group G (type 41). Inpatients with gastroenteritis and confirmed enteral adenoviruses ranged in age from one through 16 months, with a median age of seven months. Enteral adenoviruses apparently are endemic in this locale, as one or more of these viruses have been found in every calendar month for nine successive years. PMID- 2982961 TI - Identification of respiratory virus antigens in middle ear fluids of children with acute otitis media. AB - Antigens of respiratory viruses were tested by immunoassay of the middle ear fluids and nasopharyngeal secretions of 137 children with acute otitis media. The following were found: (1) an epidemic of infection with respiratory syncytial virus (RSV) caused a significant increase in the occurrence of acute otitis media. (2) Fifteen percent of the children had RSV antigens in middle ear fluid, and in 7% RSV was the sole pathogen found. Adenovirus antigens were found in the middle ear fluid of 3% of the children. (3) Bacteriologic findings in otitis media related and unrelated to viral (RSV) infection were similar. These findings indicate that some episodes of otitis media are associated with viral infection and that the disease may be solely due to viral infection. Thus, at least during an epidemic of respiratory virus infections, treatment failures--e.g., fever and earache unresponsive to antimicrobial therapy--may be due to a viral etiology of acute otitis media. PMID- 2982962 TI - Virus-induced leukopenia: Colorado tick fever as a human model. AB - Eight patients with Colorado tick fever were studied to determine whether alterations in the production of granulopoietic stimulatory or inhibitory factors (or both) could be found in association with the leukopenic state of the disease. The studies demonstrate that in the patients with Colorado tick fever the mononuclear cell production of colony-stimulating factor is decreased and that there is an increase in circulating inhibitory factors in the serum of such patients. The depressed mononuclear cell colony-stimulating activity does not appear to be reversible by addition of either endotoxin or normal human serum. Characterization of these serum inhibitory factors may facilitate understanding of leukopenia in human disease. PMID- 2982963 TI - The relation of viral replication to interstitial pneumonitis in murine cytomegalovirus lung infection. AB - Using a murine model of murine cytomegalovirus (MCMV) interstitial pneumonitis, we examined the relation between the virus content of the lung and lung disease. While MCMV alone does not cause lung disease, interstitial pneumonitis was present in all mice receiving both MCMV and a single dose of cyclophosphamide. In this case the severity of disease, judged by increases in wet weight of the lung, was proportional to the virus content of the lung. Although both acyclovir (50 mg/kg per day) and passive antibody administration reduced the MCMV titers in lung tissues by greater than 90%, histological evidence of interstitial pneumonitis was present in all animals. However, both virus inhibitors reduced the severity of interstitial pneumonitis in treated mice. While transient alterations in host immunity are necessary to induce interstitial pneumonitis after MCMV infection, the severity of interstitial pneumonitis seems to reflect the burden of virus replication. Reduction of virus growth does not prevent, but may moderate, MCMV interstitial pneumonitis. PMID- 2982964 TI - Epstein-Barr virus infections in homosexual men with chronic, persistent generalized lymphadenopathy. AB - The status of infection by the Epstein-Barr virus (EBV) in 20 patients with chronic, persistent generalized lymphadenopathy was evaluated with use of three parameters: antibodies to EBV, EBV excretion, and EB virocytemia. Results were compared with those obtained from two groups of control subjects: patients with infectious mononucleosis and healthy EBV-seropositive adults. Profiles of antibody to EBV and the prevalence of EBV excretion were essentially similar for study subjects and healthy control subjects, but the level of EB virocytemia was significantly higher in study subjects than in healthy control subjects. PMID- 2982965 TI - IgG-mediated viral clearance in experimental infection with herpes simplex virus type 1: role for neutralization and Fc-dependent functions but not C' cytolysis and C5 chemotaxis. AB - For determination of whether the Fc moiety is required for antibody effectiveness in models of herpes simplex virus type 1 (HSV-1) infection, the effects of immune IgG and F(ab')2 fragments were compared by using a passive transfer model of footpad infection. In the IgG- and the F(ab')2-treated groups illness developed in 2 (10%) of 20 and 6 (25%) of 24 mice, respectively, compared with 10 (63%) of 16 controls. IgG treatment markedly, and F(ab')2 treatment moderately, reduced footpad viral titer and viral spread to sciatic nerve and spinal cord. The marked viral clearance by IgG was not attributable to C'-dependent lysis because rapid viral clearance was observed in C5-deficient B10.D2/oSn mice. Viral latency as a consequence of acute infection occurred in 38 (63%) of 60 lumbosacral dorsal root ganglia in the control group, 5 (8%; P less than .001) of 60 in the IgG-treated group, and 26 (33%; P less than .01) of 78 in the F(ab')2-treated group. PMID- 2982966 TI - Cross-resistance to nalidixic acid, trimethoprim, and chloramphenicol associated with alterations in outer membrane proteins of Klebsiella, Enterobacter, and Serratia. AB - We studied in vitro mutants of Klebsiella, Enterobacter, and Serratia cross resistant to nalidixic acid, trimethoprim, and chloramphenicol that were similar to mutants found in vivo. The sole mechanism for this type of resistance appeared to be a reduction in permeability of the cell envelope. The mutants had significantly lower rates of uptake of glucose and chloramphenicol, but binding of chloramphenicol to ribosomes was normal. In addition, the amounts of dihydrofolate reductase were similar in both wild-type and cross-resistant mutants of Klebsiella. Examination of the bacterial outer membrane revealed that the amount of at least one major protein, with a molecular size of approximately 40 kilodaltons, was decreased in the mutants. Therefore the resistance seemed likely to be due to the reduction in quantity of these outer membrane proteins, possibly porins, in the mutant bacteria. PMID- 2982967 TI - Herpes simplex virus replication in interleukin-2-stimulated human T cells. PMID- 2982968 TI - Acute and latent infection by thymidine kinase mutants of herpes simplex virus type 2. PMID- 2982970 TI - A statewide assessment of the role of Norwalk virus in outbreaks of food-borne gastroenteritis. PMID- 2982969 TI - Detection of two human T cell leukemia/lymphotropic viruses in cultured lymphocytes of a hemophiliac with acquired immunodeficiency syndrome. PMID- 2982971 TI - The long-term serological course of asymptomatic hepatitis B virus carriers and the development of primary hepatocellular carcinoma. AB - One hundred fifty asymptomatic patients who were carriers of hepatitis B surface antigen (HBsAg) were studied serologically for up to 11.3 years (mean, 6.1 years). Only 9 (6.0%) lost HBsAg during the study period, for a mean annual clearance rate of 1.0%. We found no difference in the clearance of HBsAg by age, but a higher percentage of females lost HBsAg than did males (P less than .02). Hepatitis B e antigen (HBeAg) was found in 102 (68.5%) of the 149 carriers of HBsAg who were tested. Carriers of HBsAg who were seropositive for HBeAg were younger than those who were seronegative for HBeAg (P less than .01). The prevalence of HBeAg was not affected by the patients' sex. The clearance of HBeAg was gradual; 9.6% of the HBsAg carriers lost HBeAg each year. Females were more likely to clear HBeAg than were males (P less than .01), and those who cleared HBeAg were older than those who did not (P less than .01). Three (2.0%) of the HBsAg carriers developed primary hepatocellular carcinoma during the study period. PMID- 2982972 TI - Intranasal interferon-alpha 2 prophylaxis of natural respiratory virus infection. PMID- 2982973 TI - Resistance of seropositive volunteers to subcutaneous challenge with low-passage human cytomegalovirus. PMID- 2982974 TI - Fomites and herpes simplex virus. PMID- 2982976 TI - [A study on alkaline phosphatase in cervical mucus using disc electrophoresis]. AB - A simple filter-disc-absorption technique for sampling human cervical mucus had been developed by colleagues of author. Using this technique, electrophoretic patterns of proteins and phosphorylase in cervical mucus had been reported. In this presentation, I report isoenzyme patterns of alkaline phosphatase (ALP) in the cervical mucus of normal pregnant women and patients with various gynecological diseases using this developed technique. Electrophoresis was carried out with 11.25% polyacrylamide separating gel. The separating gel was prepared using the stacking buffer system at pH 6.7. By this system, the stacking effect was maintained in the gel and the molecular sieve effect was sharpened. ALP activity was demonstrated using 5-bromo-3-indolyl phosphate as the substrate. Placental ALP was identified by its electrophoretic mobility and thermostability. Placental ALP was demonstrated in cervical mucus from 85 pregnant women as early as 6 weeks' gestation. In sera, however, the enzyme activity was demonstrated after 21 weeks' gestation. In cervical mucus and sera of non-pregnant women, and of patients with myoma of the uterus, ovarian tumor and cervical cancer, placental ALP was not demonstrated. PMID- 2982975 TI - [Search for possible routes of vertical and horizontal transmission of adult T cell leukemia virus]. PMID- 2982977 TI - [A light and electron microscopic study of papillary hidradenoma of the vulva]. AB - Papillary hidradenoma is a rare benign tumor, which occurs mostly on the labia major or in the perianal region. Two cases were studied by light and electron microscopy. Light microscopically, these lesions were seen to consist of papillary adenomatous structures, whose lumina were lined by two to three layered cuboidal epithelial cells. Nuclear atypia and mitotic figures of these epithelial cells were rare. Nipple-like projections of their eosinophilic cytoplasm toward the lumina, resembling those of "apocrine" secretion, were frequently observed. Electron microscopy revealed that the epithelial cells lining the lumina had many secretory granules, lysosome-like structures with lipid, and large atypical mitochondria in their cytoplasm. The apical portion of the cytoplasm, which contained numerous secretory granules, often protruded toward the lumina. These ultrastructural features were considered to be similar to those of apocrine gland secretory cells. Moreover, in the basal layer of epithelium, spindle-shaped cells with features of typical myoepithelial cells were identified. These light and electron microscopic findings suggest that papillary hidradenoma of the vulva is one of the tumors of skin appendages with direction of differentiation toward the apocrine glands. PMID- 2982978 TI - [Studies on specific receptors for prolactin (PRL) in the rat ovary]. AB - The characteristics of specific receptors for prolactin (PRL) in the ovary were examined. The binding of 125I-ovine PRL to the receptor was dependent upon time, temperature, and tissue concentration. Equilibrium was reached by 16 h at 23 degrees C in the 12.5 mg tissue/tube. Scatchard analysis revealed a high affinity, low capacity and single class of binding site. Ovine PRL bound to rat liver membranes with a Kd (dissociation constant) of 3.51 X 10(-10) M and to rat ovarian membranes with a Kd of 2.21 X 10(-10) M, respectively. The number of binding sites in rat liver and ovary were 205.2 fmoles/mg of protein and 226.1 fmoles/mg of protein, respectively. Bound 125I-ovine PRL was displaceable by unlabelled human, ovine and rat PRL but not by rat LH, FSH, TSH or GH. However, on the immunological aspects, 125I-ovine PRL bound only to anti-ovine PRL serum but did not bind to anti-human PRL serum or anti-rat PRL serum. Unlabelled human and rat PRL could not inhibit the binding of labelled ovine PRL to the anti-ovine PRL serum. These data indicate the existence of specific PRL receptors in the rat ovary, the characteristics of which are similar to those of receptors in the liver. The binding specificity for various mammalian species suggests a difference between biological and immunological binding sites of the PRL molecule. PMID- 2982979 TI - Myeloperoxidase-induced inactivation of alpha 1-antiprotease in hamsters. AB - While numerous studies have demonstrated that the myeloperoxidase system found in neutrophils can oxidize and functionally inactivate alpha 1-proteinase inhibitor in vitro, there is little direct evidence that this phenomenon is relevant in vivo. Using incubation with tritiated porcine pancreatic elastase followed by column chromatography to quantitate binding, we demonstrated recovery of microgram amounts of functional alpha 1-protease inhibitor from bronchoalveolar lavage of hamster lungs. When exposed to the myeloperoxidase system in vitro, hamster alpha 1-protease inhibitor was 97% inactivated. Functional alpha 1 protease inhibitor recovered by bronchoalveolar lavage 20 minutes after hamsters were given intratracheal injections with myeloperoxidase and either hydrogen peroxide or glucose plus glucose oxidase was only half that recovered from control animals. These studies suggest that the myeloperoxidase system is effective in oxidizing alpha 1-protease inhibitor in vivo. They support the concept that oxidation of alpha 1-protease inhibitor by myeloperoxidase from neutrophil granules in the presence of H2O2 and halide may produce elastase antielastase imbalance in vivo and contribute to the development of acute lung injury and emphysema in humans. PMID- 2982980 TI - Unresponsiveness of plasma mineralocorticoids to angiotensin II in diabetic patients with asymptomatic normoreninemic hypoaldosteronism. AB - offlated hypoaldosteronism with or without hyperkalemia in patients with diabetes mellitus has been shown to exist occasionally without hyporeninemia. To assess in detail the adrenal function in this disorder, the responses of plasma aldosterone (PA) and its precursor steroids to angiotensin II (AII) infusion and adrenocorticotropic hormone (ACTH) injection were studied in seven patients with asymptomatic normoreninemic hypoaldosteronism (ANH) and 11 age-matched normal subjects. The ANH diabetic patients had, by definition, a low PA level after furosemide (80 mg orally) plus upright posture (4 hours) stimulation, low PA and high plasma renin activity (PRA) increases after the stimulation (a low delta PA/delta PRA ratio), and normokalemia. Plasma inactive renin and the inactive renin/total renin ration were similar in the ANH diabetic patients and in the normal subjects. Under the pre-AII condition, plasma DOC and corticosterone levels tended to be low, and the plasma 18-OHB and PA levels were low in the ANH diabetic patients compared with the normal subjects. The ratio of plasma 18-OHB to PA was similar in the two groups. All infusion produced no increases in plasma 18-OHB and PA in the ANH diabetic patients, whereas the infusion caused dose dependent increases in these steroids in the normal subjects. Plasma DOC and corticosterone levels remained unchanged during AII infusion in the two groups. ACTH injection produced appropriate PA increases relative to the basal PA in the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982981 TI - Hypochlorite radioiodination of parathyroid peptides (hPTH1-34, [Tyr43]hPTH44 68). AB - Parathyroid hormone (PTH) radioimmunoassays have conventionally utilized [125I]bPTH1-84 as a radioligand, but more region-specific PTH assays are now possible with the use of recently available synthetic PTH peptides as standards and radioligands. A radioiodination procedure has been developed that utilizes hypochlorite as an oxidant and that is capable of producing PTH tracers of high specific activity (200 to 250 microCi/micrograms), prolonged stability, and excellent immunologic potency. Radioiodinated hPTH1-34 and [Tyr43]hPTH44-68 produced by hypochlorite iodination techniques can be used to develop sensitive and region-specific PTH assays of use in clinical and research situations. PMID- 2982983 TI - Synovial sarcoma of the hypopharynx. PMID- 2982982 TI - Clinical and laboratory investigation of the effects of epsilon-aminocaproic acid on hemostasis. AB - We previously reported that epsilon-aminocaproic acid (EACA) prolonged the bleeding time in patients with intracranial aneurysms when given in doses of 36 to 48 gm/day. We now show that doses of 24 gm/day also prolong the bleeding time, but only after 72 hours of continuous infusion. The effect on the bleeding time correlates with the duration of EACA therapy but not with the plasma level of the drug. Bleeding times return toward normal within 72 hours of discontinuing EACA infusions. The factors responsible for the bleeding time prolongation were investigated. In vitro, EACA inhibited adenosine diphosphate- and collagen induced platelet aggregation and the release of platelet adenosine triphosphate and serotonin. It also prevented the adenosine diphosphate-stimulated binding of fibrinogen to intact as well as to chymotrypsin-treated platelets. However, platelets obtained from patients who had received EACA showed little functional impairment. This observation indicates that the focus of EACA activity in vivo is probably not the platelet per se, but the platelet-vessel wall interaction or a vascular component alone. EACA did not enhance prostacyclin production or release from cultured bovine endothelial cells. The fact that the effects of the drug on the bleeding time were related to the duration of EACA therapy suggests that an accumulation of the drug on the vessel wall may be required before alterations in hemostasis are observed. EACA in a daily dose of 24 gm significantly impaired fibrinolysis, but supranormal levels of fibrinolytic activity were observed within 72 hours of stopping the drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982984 TI - Primary oat cell carcinoma of the larynx. AB - A case of oat cell carcinoma of the epiglottis eleven years after radiotherapy to the larynx for squamous cell carcinoma of the vocal cords is presented. There is a 13.5 per cent incidence of a second neoplasm in these cases. This is a very aggressive tumour and a combination of surgery, radiotherapy and multi-drug chemotherapy appears to offer the best hope of cure. The two-year survival rate is only 8.5 per cent. PMID- 2982985 TI - Modulation of mononuclear phagocyte cytotoxicity by alpha-tocopherol (vitamin E). AB - The in vitro effect of colloidal suspensions of alpha-tocopherol (alpha-T) on phorbolmyristate-acetate (PMA)-induced monocyte cytotoxicity and on antibody dependent monocyte cytotoxicity (ADCC) was studied. We observed that 1) in the presence of alpha-T, the inhibition was twice as high in the PMA-induced assay than in ADCC; 2) monocytes preincubated with alpha-T were inhibitory in both assays but much less in ADCC, and 3) target erythrocytes preincubated with alpha T decreased the cytotoxicity in the PMA-induced assay only. Since alpha-T preincubated monocytes showed a decreased release of H2O2 but not of O2-, we concluded that one of the mechanisms by which alpha-T decreased cytotoxicity could be decreased release of H2O2. Whereas the role of H2O2 was documented in the PMA-induced cytotoxicity, in ADCC non-oxidative injury seems more important. This is supported by 1) lesser inhibition of the assay with alpha-T preincubated monocytes; 2) lack of protection with alpha-T preincubated erythrocytes, and 3) mild inhibition with protease inhibitor. PMID- 2982986 TI - Relationship between adrenocorticotrophin bioactivity in blood and cerebrospinal fluid of rhesus monkeys. AB - Adrenocorticotrophin levels, measured by a cytochemical bioassay, were determined in the plasma and cerebrospinal fluid (CSF) of adult female rhesus monkeys which were ovariectomized and receiving oestrogen replacement therapy. In control monkeys, ACTH bioactivity was found in both CSF (10.2 +/- 1.8 ng/l) and plasma (186 +/- 51 ng/l) in samples taken at 14.00 h (lights on: 07.00-19.00 h). Dexamethasone treatment (0.2 mg/kg) twice daily for 4 days suppressed plasma ACTH levels (52.8 +/- 25.2 ng/l) but had no effect on CSF levels (7.6 +/- 2.7 ng/l). Raising plasma ACTH, either by daily injections of a long-acting preparation of ACTH (1-24) for 6 days or by bilateral adrenalectomy (and subsequently with drawing cortisol replacement therapy) also resulted in no detectable changes in ACTH levels in the CSF. A regression analysis between ACTH in the plasma and CSF from samples taken throughout the experiments showed no correlation. In contrast, measurement of ACTH by radioimmunoassay, whilst satisfactory for determination of this peptide in plasma, could not identify authentic ACTH in the CSF. It is concluded that bioactive ACTH does not enter the CSF in detectable quantities from either the peripheral vascular compartment or from the animal's own pituitary gland, and that reducing ACTH secretion from the pituitary also has no effect on levels of ACTH in the CSF. This is in marked contrast to other pituitary peptide hormones, including prolactin, which is secreted together with ACTH during 'stress' but which, unlike ACTH, enters the CSF relatively easily. PMID- 2982987 TI - Characteristics of bovine luteal cells in culture: morphology, proliferation and progesterone secretion in different media and effects of LH, dibutyryl cyclic AMP, antioxidants and insulin. AB - The morphological and functional characteristics of cultured bovine luteal cells were examined. Dispersed luteal cells were cultured in either Ham's F12 medium or Dulbecco's Modified Eagle's Medium (DMEM), plus 10% adult bovine serum. After 18 h of culture, fibronectin was observed to be associated with some cells showing histochemical 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity. In DMEM, cells adopted a fibroblast-like appearance with a doubling time of 34 h while in Ham's F12, cells retained an epithelial-like morphology with a doubling time of 91 h. Dibutyryl cyclic AMP (dbcAMP) but not LH slowed proliferation and altered cell morphology in DMEM. Addition of insulin and dbcAMP was required to maintain 3 beta-HSD activity in cells cultured for more than 3 days. Progesterone secretion was significantly greater in Ham's F12 than in DMEM although there was a rapid decline in basal secretion during culture in either medium. Both LH and dbcAMP stimulated progesterone secretion although the effects of dbcAMP were significantly greater than those of LH after 18 h of culture. Addition of insulin significantly enhanced basal, LH-stimulated and dbcAMP-stimulated progesterone secretion after 3 days of culture whereas several antioxidants were without effect. These studies indicate that the morphological and functional characteristics of cultured luteal cells may be best maintained in Ham's F12 plus dbcAMP and insulin. PMID- 2982988 TI - Control of zona glomerulosa function in the isolated perfused rat adrenal gland in situ. AB - The extent to which results obtained using in-vitro techniques can be taken to reflect in-vivo physiological responses in the study of adrenocortical function has not been subjected to systematic study. Some evidence suggests that in-vitro preparative methods may affect the secreted steroid profile. For this reason it seemed desirable to study adrenal function using an isolated perfused whole gland technique, and this study reports results obtained with known aldosterone stimulants. Angiotensin II, ACTH and potassium ions all stimulated aldosterone secretion in a dose-dependent manner. The stimulation thresholds of these substances were compatible with their normal circulating concentrations. For angiotensin II stimulation this preparation was two orders of magnitude more sensitive than any in-vitro preparation. Most importantly, the specific glomerulosa effectors, angiotensin II and potassium, selectively stimulated aldosterone output, and had no consistent effect on corticosterone secretion at any dose used. On the other hand, ACTH stimulated both corticosterone and aldosterone output at all effective concentrations. The actions of alpha-MSH were also studied using this preparation. Low doses of alpha-MSH selectively stimulated aldosterone secretion, while higher doses were needed to stimulate corticosterone. The onset of response to all stimulants was invariably seen within the first 10 min after administration of stimulants. Maximal aldosterone output was achieved within the first 10 min whereas corticosterone secretion usually peaked 10-20 min later. The amount of aldosterone produced by this preparation was much higher than the amount produced by dispersed cell preparations, and closely approximated to the levels of aldosterone obtained in adrenal vein blood.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2982989 TI - Antihemostatic, antiinflammatory, and immunosuppressive properties of the saliva of a tick, Ixodes dammini. AB - Pilocarpine-induced saliva of the tick, Ixodes dammini, inhibited platelet aggregation triggered by ADP and collagen, as well as platelet-aggregation factor. In addition, we found apyrase activity (which degrades ATP and ADP to AMP and orthophosphate) and an anticoagulant. We showed the presence of prostaglandin E2 (PGE2) by bioassay and radioimmunoassay. This saliva inhibited interleukin 2 production by T cell hybridomas, an activity consistent with that of PGE2. A kininase was demonstrated, and this may counteract the algesia- and edema promoting properties of PGE2. Together, these salivary components produce antihemostatic, antiinflammatory, and immunosuppressive effects that may facilitate feeding, as well as transmission of tick-borne pathogens. PMID- 2982990 TI - Serum factor requirement for reactive oxygen intermediate release by rabbit alveolar macrophages. AB - Alveolar macrophages (AM) from pathogen-free rabbits were unable to release reactive oxygen intermediates (ROI) unless they were conditioned in serum for 24 48 h before triggering with membrane-active agents. The degree of serum conditioning of AM depended upon the concentration of serum used; optimal ROI release was obtained at or above 7.5% fetal bovine serum (FBS). FBS, autologous rabbit serum, pooled rabbit serum, and pooled human serum were each capable of conditioning AM for release of ROI. Serum conditioning of AM requires synthesis of new protein(s); and the enzyme required for ROI production, NADPH oxidase, was only detectable in serum-conditioned cells. Moreover, serum-conditioned cells lost their ability to release ROI after transfer to serum-free medium, while cells maintained in serum-free medium acquired the capacity to release ROI after their transfer to serum-containing medium, demonstrating the reversibility of the phenomenon. Initial purification data indicate that conditioning is mediated by a discrete serum constituent, which precipitates 40-80% saturated ammonium sulfate, does not bind to Cibacron Blue columns, and has a molecular weight of 30,000 to 50,000, as determined by molecular exclusion chromatography. Unlike gamma interferon, which also enhances ROI release by macrophages, our serum conditioning factor is not acid labile, retaining 67% of its activity after 120 min incubation at pH 2.0. Moreover, it does not appear to be a contaminating endotoxin, since LPS neither conditioned AM for ROI production, nor triggered ROI production by serum-conditioned AM. We propose that such a conditioning requirement may normally protect the lung against ROI-mediated tissue injury. However, during a pulmonary inflammatory reaction initiated by other mediator systems, the resulting transudation of plasma proteins into the alveolar spaces may condition AM in situ for ROI production. PMID- 2982991 TI - Abnormalities induced by the mutant gene, lpr. Patterns of disease and expression of murine leukemia viruses in SJL/J mice homozygous and heterozygous for lpr. AB - SJL/J mice heterozygous or homozygous for the lpr mutation were compared with SJL/J-+/+ mice for longevity, histopathology, antigenic characteristics of lymphocytes and expression of murine leukemia viruses (MuLV). In comparison to +/+ mice, lpr homozygotes had a markedly shortened life span, died with infiltrative pulmonary disease, but little or no renal disease, and expressed high levels of infectious ecotropic MuLV in lymphoid tissues. SJL-lpr/+ mice had a life span intermediate between SJL-+/+ and -lpr/lpr mice, died with lymphomas that histologically resembled the neoplasms of +/+ mice, and sometimes expressed high levels of ecotropic MuLV. The lymphomas of lpr/+ could be transplanted to +/+ recipients in 78% of cases, and continuous in vitro lines were established from some of them. Similar effects on virus expression or lymphoma development were not observed in other strains homozygous or heterozygous for the lpr mutation. These results indicate that the diseases expressed by mice homozygous for the lpr mutation are highly strain-dependent, and that this gene can have an effect in the heterozygous state in SJL mice. PMID- 2982992 TI - Interaction of the heterozygous nude gene with the asplenia trait in mammary tumorigenesis. AB - The BALB/c mouse strain has been shown to contain endogenous mouse mammary tumor virus (MMTV) proviral sequences. However, no exogenous MMTV particles have been detected in their tissues. Female BALB/c mice from our colonies exhibit a very low incidence of spontaneous mammary tumors (SMT); less than 1% at up to 20 mo of age. Immunodeficient BALB/c mice heterozygous for the nude gene (nu/+, +/+), for the dominant hemimelia gene associated with asplenia (+/+, Dh/+), or for both traits (nu/+, Dh/+) have been examined for SMT incidence and the presence of MMTV proviruses. Based on restriction digestion with Eco RI, Bam HI, and Pst I, the immunodeficient mice have an MMTV provirus copy number and organization identical to the BALB/cCrgl strain. This MMTV DNA pattern is distinct from the MMTV proviruses in C3H/He, C57BL/6J and CBA/CaJ mice, which were parental strains of the immunodeficient mutants. Normal female BALB/c or BALB/c heterozygous for the asplenic trait do not develop significant numbers of SMT at up to 19 mo of age. In contrast, an incidence of 23.8% and 57.7% SMT was observed in BALB/c nu/+ heterozygotes, and in BALB/c nu/+, Dh/+ heterozygotes, respectively. These results indicate that agenesis of the spleen, concomitant with the presence of the heterozygous nude gene, contribute to a high incidence of SMT in the low-SMT BALB/c mouse strain. PMID- 2982993 TI - Lethal factors and enzymes in the secretion from Duvernoy's gland of three colubrid snakes. AB - Secretion from Duvernoy's gland of the colubrid snakes Malpolon, Spalerosophis, and Thamnophis was obtained by pilocarpine stimulation and tested for lethality and selected enzymatic activities. Pools of secretion from Malpolon and Spalerosophis were fractionated by gel filtration, and several major active fractions were examined. The secretion from Malpolon had an LD50 of 6.5 micrograms/gm in mice; two lethal fractions with LD50's of 2.75 micrograms/gm and 4.5 micrograms/gm were isolated. One of these fractions appears to be a basic phospholipase A with a molecular weight of about 17,000. Spalerosophis had a secretion with an LD50 of 2.75 micrograms/gm in mice; one main lethal fraction with an LD50 of 2.5 micrograms/gm was isolated. Secretion from Thamnophis had an LD50 of 33.3 micrograms/gm; it was not fractionated owing to a paucity of material. At least two of the snake species examined have toxic secretions that may be important during feeding by killing or weakening and helping to subdue the struggling prey. PMID- 2982994 TI - Nerve growth factor induces neural differentiation in undifferentiated cell of early chick embryos. AB - Nerve growth factor (NGF) induced differentiation in postnodal pieces (PNPs) of stage 4 chick embryos. This induction was highly selective for neural tissue; no other structures developed in the NGF-treated PNPs. Furthermore, the number of PNPs showing neural differentiation was dependent on the concentration of NGF, but there was no correlation between the concentration of NGF (5-100 ng/ml) and extent of neuralization. The neural inducing capacity of NGF could be abolished by anti-NGF antibody. NGF-induced neural differentiation was accompanied by elevated intracellular levels of cyclic AMP. Exogenous cyclic AMP (175 micrograms/ml) was able to stimulate neural differentiation but, unlike NGF, induced other structures (e.g., notochord and pulsatile tissue). Overall results suggest that cells from chick embryos at developmental stages much earlier than previously thought are responsive to NGF and NGF or a a closely related substance may serve as a neural inducer in the chick embryo. PMID- 2982995 TI - Peroxidase activity in the epithelium of the digestive tract of the bullfrog, Rana catesbeiana. AB - Peroxidase activity was examined cytochemically in the mucosal epithelium along the length of the digestive tract from the esophagus through the large intestine during the development of the bullfrog, Rana catesbeiana. In the tadpole of this species, cells with peroxidase activity were found abundantly in the esophagus, stomach, and large intestine; and the types of such cells differed according to the region: ciliated cells and mucous cells in the esophagus; ciliated cells in the stomach; and brush cells, absorptive cells, and goblet cells in the large intestine, respectively. After metamorphosis, however, peroxidase activity was observed exclusively in absorptive cells and goblet cells in the large intestine. Peroxidase activity was commonly demonstrated in apical vesicles or granules, to some degree in rough endoplasmic reticulum, and in some elements of the Golgi apparatus. Furthermore, reaction product was also found in mucus covering the luminal surface of such epithelial cells. These findings indicate that peroxidase positive cells, which may have the ability to synthesize peroxidase as a secretory product, were distributed mainly in three regions of the digestive tract in tadpoles (esophagus, stomach, and large intestine), but were centered in one specific region, the large intestine, after metamorphosis. Concomitantly, the variety of types of peroxidase-positive cells decreased during metamorphosis. Our results indicate that some of the peroxidase in the digestive tract may have a secretory origin and may play a role in the defense against microorganisms. PMID- 2982996 TI - Possible function of 17 beta-hydroxysteroid dehydrogenase in preimplantation hamster embryos. AB - 17 beta-Hydroxysteroid dehydrogenase (17 beta-HSD) catalyzes the interconversion of estradiol-17 beta (E2) and estrone (E1). The present study is designed to investigate the following: (1) the developmental stage of hamster embryos at which 17 beta-HSD activity first becomes detectable, and (2) the E1----E2 and E2- --E1 conversion rate in the preimplantation hamster embryo. Embryos obtained from superovulated hamsters on days 1-4 were cultured in medium containing 107 ng [3H]E1 or -E2/ml and the respective conversion product, [3H]E2 or -E1, was isolated and assayed. The results show that (1) E1----E2 conversion was active in all embryos at the rate of 0.57, 0.66, 0.54 and 0.48 fmol/embryo/hr for day 1 (one-cell), 2 (two-cell), 3 (eight-cell) and 4 (blastocyst), respectively, and (2) E2----E1 conversion was not detectable in hamster embryos. In long-term blastocyst culture, E2----E1 conversion becomes detectable at 25 hours and increases sharply from 25 to 47 hours. These results suggest that (1) 17 beta-HSD may function mainly to convert E1 into E2 in preimplantation hamster embryos and (2) E2----E1 conversion may become active only during and after implantation. PMID- 2982997 TI - Extracellular localization of cyclic GMP in the house cricket male accessory reproductive gland and its fate in mating. AB - The male accessory reproductive gland (ARG) of the house cricket, Acheta domesticus (L.), contains an exceedingly high concentration of cyclic GMP, about 1,000 pmol/mg protein. Immunofluorescent localization and radioimmunoassay measurements show that cyclic GMP is concentrated in a small number of tubules. It accumulates in the tubule lumina where it is protected from degradation by phosphodiesterases. Cyclic GMP is secreted by the ARG and is incorporated into spermatophores. Over 80% of spermatophore cyclic GMP is found in the handle capillary tube, a thin conduit through which sperm pass during transfer to the female. The concentration of cyclic GMP in the insemination fluid is about 20 microM but does not appear to be specifically associated with the sperm. Cyclic GMP enters the female spermatheca during insemination but disappears rapidly. Physiological effects of cyclic GMP on sperm were not observed nor was an effect of cyclic GMP observed on egg laying by mated females. Cyclic AMP was localized on sperm flagella in the spermatophore and in the spermatheca. These studies indicate that cyclic nucleotides have important roles in insect reproduction and that the house cricket is a good model for elucidating these functions. PMID- 2982998 TI - Semliki Forest virus infection of mice: a model for genetic and molecular analysis of viral pathogenicity. PMID- 2982999 TI - Cytomegalovirus infection of human teratocarcinoma cells in culture. AB - Whereas human cytomegalovirus (HCMV) did not replicate in human embryonal carcinoma (EC) cells, it did replicate in some of the differentiated cells arising following the exposure of TERA-2-derived human EC cells to retinoic acid. On the other hand, retinoic acid did not induce a permissive state in several other diverse human cell lines, including an EC line, 2102Ep, which did not differentiate in response to this agent. Also, both TERA-2 and 2102Ep EC cells differentiated to a limited extent when grown at low cell density and a few of these cells became permissive for HCMV. Thus, susceptibility is the result of differentiation and not due to a direct effect of retinoic acid on viral replication. The nature of the block to HCMV replication in human EC cells is unknown, but viral DNA could be detected in the nucleus within an hour of infection and there was an increased anchorage-independent growth of undifferentiated and differentiated cells following HCMV infection. Viral replication is not subject to a general block in these cells, since another herpesvirus, herpes simplex virus type 1, replicated well. PMID- 2983000 TI - Herpes simplex virus sequences involved in the initiation of oncogenic morphological transformation of rat cells are not required for maintenance of the transformed state. AB - We have determined the herpes simplex virus (HSV) type 2 DNA sequences responsible for the initiation of morphological transformation and have investigated the retention and expression of these sequences in morphologically transformed cells and in tumours derived from these cells. All the transformed cells analysed were selected by a focus formation assay and are oncogenic in the inbred host rat. Cloned HindIII and Bg/II fragments from the HSV-2 genome were assayed for the ability to initiate morphological transformation of rat embryo cells. Only the HindIII a (map units 0.52 to 0.72) and the Bg/II n (0.582 to 0.612) clones gave transformed foci. This shows that the Bg/II n region is responsible for initiation of transformation. Southern blot analysis of DNA extracted from these transformed cells and from tumours derived from these transformed cells revealed that neither the Bg/II n fragment nor fragments of 500 bp mapping within it are detected at the level of one copy per cell and therefore need not be retained in the cell to maintain the oncogenic phenotype. In addition there was no evidence of expression of the HSV-specified ribonucleotide reductase activity which is partially encoded within the Bg/II n fragment of HSV-2. We also analysed DNA from rat embryo cells transformed by ts mutants of HSV-2 (HG52) or HSV-1 (HFEM or 17) at non-permissive temperature or by virus at supraoptimal temperature or by sheared virus DNA and DNA from tumours derived from lines of these transformed cells. In addition, we cloned both transformed and tumour cell lines and analysed these similarly. In no case could we detect HSV DNA sequences at the level of one copy per cell. PMID- 2983001 TI - Sequences of the nucleocapsid genes from two strains of avian infectious bronchitis virus. AB - cDNAs prepared from viral genomic RNA purified from two strains of infectious bronchitis virus (IBV) (Beaudette and M41) have been cloned into pBR322. Three of these clones, which contain the complete sequences of mRNA A for both strains, except for the leader sequences which are only present on the subgenomic messenger RNAs, have been sequenced using the dideoxy method. The sequences are similar for both strains, each containing a single long open reading frame of 1227 bases which predicts a polypeptide of molecular weight approximately 45 000. The genome position and size of this predicted polypeptide are consistent with it being the gene for the nucleocapsid protein. The amino acid sequence shows considerable homology with those of the nucleocapsids of murine hepatitis virus strains A59 and JHM. The major difference between the sequences determined for the two IBV strains is that the 3' non-coding region of the Beaudette strain contains a 184 base segment which is not present in the M41 strain. PMID- 2983002 TI - Peptide mapping of envelope-related glycoproteins specified by the flaviviruses Kunjin and West Nile. AB - Glycoproteins detected in Vero cells infected by the flaviviruses West Nile and Kunjin were examined by gel electrophoresis and peptide mapping. Two major glycoproteins, gp66 and gp54, were observed in West Nile virus-infected cells labelled for short time periods with [3H]mannose. A third glycoprotein, gp58, was present in smaller amounts. Pulse-labelling experiments suggested that gp66 was a precursor of gp54. Peptide mapping of [3H]leucine-labelled gp66, gp54 and the envelope glycoprotein E of West Nile virus demonstrated that gp66 and gp54 were related to E, and that the peptides of gp54 were a subset of those of gp66. Peptide mapping of the corresponding Kunjin virus-specified glycoproteins (gp66, gp59 and gp53) showed that the [3H]leucine-labelled peptides of gp53 and gp59 were similar and were contained within gp66. Since we have shown previously that gp59 and gp53 are related to E of Kunjin virus, we conclude that cells infected by West Nile or Kunjin viruses contain a similar set of E-related glycoproteins. PMID- 2983003 TI - Lack of quantitative correlation between the neutralization of poliovirus and the antibody-mediated pI shift of the virions. AB - The effect of mono- and polyclonal antibodies on the infectivity and pI (isoelectric pH) of type 1 poliovirus was studied. According to Mandel's hypothesis, the isoelectric pH of poliovirus should change to about pI 4 upon neutralization. However, several antibodies did not follow this rule. Moreover, when antibodies did shift the pI, no quantitative correlation existed between the extent of neutralization and the amount of poliovirus shifted to low pI. PMID- 2983004 TI - Human papillomavirus type 16 recombinant DNA is maintained as an autonomously replicating episome in monkey kidney cells. AB - Human papillomavirus type 16 (HPV16) DNA cloned in the expression vector pSV2-neo has been shown to be maintained in transfected monkey kidney cells as an autonomously replicating episome at a level of 2 to 10 copies per cell. Integration of pSV2-neo/HPV16 DNA with the host genome occurred in transfected mouse fibroblasts. Neither type of cell appeared to be phenotypically transformed. PMID- 2983005 TI - Recombinants between vaccinia and ectromelia viruses bearing the specific pathogenicity markers of both parents. AB - Nineteen recombinants between vaccinia virus (VV) DNA-temperature-sensitive mutants and ectromelia virus (EMV) were characterized with respect to their biological properties and genome structure. Four of these recombinants acquired the pathogenicity for mice characteristic of EMV, while the pathogenicity for rabbits characteristic of VV was not only preserved, but even enhanced. Most unexpectedly, these recombinants with 'double pathogenicity', as well as four other recombinants, acquired a stable genetic trait which was not typical of the parental viruses, i.e., the ability to form haemorrhagic lesions on the chorioallantoic membrane of chick embryos. Approximate mapping of the genomes of these recombinants with restriction endonucleases showed that their DNA contained mostly VV sequences with a single detected insert of EMV DNA. In the 'double pathogenicity' recombinants, this insert was located in the central part of the genome, and its minimal size was about 16 Mdal. PMID- 2983006 TI - Effect of heat shock on Epstein-Barr virus and cytomegalovirus expression. AB - The effect of heat shock was investigated on lymphoblastoid cell lines Raji and P3HR1 harbouring Epstein-Barr virus (EBV) genomes, and on Vero cells abortively infected with human cytomegalovirus (HCMV). A heat shock at 44 degrees C for 10 min induced the appearance of EBV early antigens in Raji cells and increased the percentage of cells expressing EBV viral capsid antigens in P3HR1 cells. Heat shock performed on Vero cells just before HCMV infection resulted in an approximately fourfold increase in the number of cells exhibiting early nuclear antigens, and in the appearance of HCMV-induced Fc receptors. PMID- 2983007 TI - Etiopathogenetic aspects of hepatitis A. I. Excretion of hepatitis A virus, biochemistry of liver function, and humoral immune response in patients with hepatitis A on admission to hospital. AB - The excretion of hepatitis A virus (HAV) in stools from 30 patients with clinically overt hepatitis A infection on the day of their admission to the hospital was determined and compared with the dynamics and values of biochemical indices of hepatocyte injury as well as with the immune response to HAV. Virus was found in 16 out of 30 stools (53%) collected within 1 week after the appearance of clinical symptoms. In sera obtained on the day of hospitalization both IgM and IgA anti-HAV were detected in all of the 30 patients, while IgG anti HAV were found in 20 (67%). There was a correlation between HAV excretion and increasing SGPT upon admission to hospital, while the level of SGPT or bilirubin as well as presence or absence of IgG anti-HAV did not correlate with excretion of HAV. HAV from stools was characterized morphologically and physicochemically. The majority of particles visualized by immune electron microscopy had electrondense appearance, while electron-lucid particles were only occasionally encountered. Isopycnic banding of HAV in CsCl revealed a broad range of densities with HAV activity. Rebanding of pooled fractions containing HAV revealed peak amounts of the virus in fractions with densities 1.32-1.33 gm/cm3. PMID- 2983008 TI - Early and late antigens of human cytomegalovirus: electroimmunodiffusion assay of numbers, relationships, and reactivities with donor sera. AB - Early antigens (EA) of human cytomegalovirus extracted from cytosine arabinoside blocked cells infected with 0.01-20 infectious units (IU)/cell were assayed with human serum by electroimmunodiffusion (EID). The number of detectable EA types increased from one to eight as the IU/cell was raised from 0.01 to 10. There was no increase in the number of EA with further increases in IU/cell, with prolonged culture, or when detergent was included in the extraction buffer. At least five of the eight EA gave reactions of identity with late-time antigens (LTA) extracted from unblocked cells at late times postinfection. In studies on a panel of sera from donors who were excreting virus and donors who were not, EID was as sensitive as conventional techniques (complement fixation and indirect hemagglutination for LTA, indirect immunofluorescence for EA) in detection of both types of antibodies from excretors but less sensitive in not detecting low levels of the antibodies in some of the sera from nonexcretors. No consistent relationships were observed between donor virological status and the numbers or types of antibodies to EA and LTA. PMID- 2983009 TI - Lytic cytomegalovirus replication and the hormones of human pregnancy. AB - The frequency of human cytomegalovirus (CMV) excretion during pregnancy denoting active infection has been demonstrated to increase as gestation advances and at term involves a significant percentage of women. This increase enhances the risk of congenital infection of the fetus. Thus it appears that some factor(s) unique to the condition of pregnancy favors susceptibility to maternal CMV infection. We designed our studies to investigate the possible association of the continuously rising levels of selected hormones and this increased susceptibility. Progesterone, 17 beta-estradiol, and cortisol were added to tissue culture media in final concentrations to match those occurring in term pregnancy serum. Two strains of human foreskin cells, one neonatal and the other fetal, were treated with either single or paired combinations of hormone-containing media. Lytic CMV replication in neonatal foreskin cells was enhanced by a maximum of 5.7-fold when these cells were treated with cortisol. Such enhancement did not occur in the fetal cells. No synergistic effects were seen when cortisol was used in combination with other hormones nor when neonatal foreskin cells were replicated for at least three generations in either single or paired combinations of hormones prior to use. Differential hormonal enhancement of CMV replication in vitro suggests a possible mechanism for the increased incidence of CMV infection observed during human pregnancy. PMID- 2983010 TI - Neurotropism of a high-passage XJ strain of Junin virus. AB - Guinea pig infection with a highly passaged XJ prototype strain of Junin virus by the intramuscular route (IM) was carried out in order to study viral tropism modification in this host. The neurotropism of this strain was demonstrated by viral isolation, meningitis, and by the presence of Junin antigens as shown by immunofluorescence. These events, not previously observed with the same lower passaged strain, revealed the appearance of neurotropism after multiple passages in guinea pigs. PMID- 2983011 TI - Diagnosis of human parvovirus infection by dot-blot hybridization using cloned viral DNA. AB - The human parvovirus can be detected in serum by the immunological techniques of immune electron microscopy (IEM), counterimmunoelectrophoresis (CIE), and radioimmunoassay (RIA). A portion of the genome of this virus has been cloned in pAT153 and used as a 32P-labelled probe in dot-blot hybridization assays to detect parvovirus DNA in serum specimens. This test proved a highly sensitive means of detecting virus in microlitre volumes of serum, giving positive results for samples containing 0.5 pg viral DNA, equivalent to 10(4) virus particles. Unlike CIE and RIA the test is not affected by the presence of parvovirus specific antibody in serum specimens, and has permitted virus to be detected in specimens obtained up to 11 days after the onset of clinical symptoms of aplastic crisis. PMID- 2983012 TI - Use of enzyme-labeled antigen for the detection of immunoglobulin M and A antibody to herpes simplex virus in serum and cerebrospinal fluid. AB - A direct enzyme-linked immunosorbent assay (ELISA that used peroxidase-labeled antigen) was developed for detection of IgM and IgA antibody to herpes simplex virus (HSV). The assay uses immuno-affinity-purified antihuman IgM or IgA antibody-coated wells of microtiter plates to separate IgM or IgA from other classes of antibody in serum or cerebrospinal fluid (CSF). The presence of specific IgM or IgA is detected by subsequent, consecutive incubation with peroxidase-labeled antigen and substrate. HSV antigen was purified by sucrose gradient centrifugation and coupled with peroxidase by the periodate method. By examining sucrose-gradient-fractionated sera the assays were shown to be specific for IgM and IgA classes of antibody. None of the sera from patients with Epstein Barr virus (n = 20), cytomegalovirus (n = 20), or varicella-zoster virus (n = 8) infection or with both rheumatoid factor and IgG antibody to HSV (n = 13) reacted positively. Only one out of 78 sera from healthy persons was positive for IgA antibody to HSV, and none for IgM antibody. All 33 patients with HSV infection developed HSV-IgA, 22 developed HSV-IgM. Of the 11 patients with primary infection, all had IgM antibody in their first sera and six had IgA antibody. The corresponding figures for the 22 patients with recurrent infection were five and nine. Furthermore, HSV-IgA antibody was found in serum and CSF of all five patients with HSV encephalitis in the second week after onset of symptoms, indicating the usefulness of the assay as a noninvasive technique for diagnosing HSV encephalitis. PMID- 2983013 TI - Pathogenesis of attenuated Junin virus in the guinea pig model. AB - The purpose of this work was to elucidate the pathogenesis of attenuated Junin virus (JV) strains in the guinea pig model. Three groups of guinea pigs were infected by the IM route with 10(3) PFU of the XJC13 and XJO-attenuated strains or with the XJ pathogenic strain of JV, respectively. Viremia was studied at 3, 5, 7, 9, 12, and 14 days postinfection (pi) (a) in serum samples of all animals and in washed cells from XJC13-infected guinea pigs by conventional techniques and (b) in whole blood samples from XJC13 and XJO animals by coculture with Vero cells. Virus spread was studied at 14 days pi in brain, spleen, lymph nodes, and bone marrow by parallel suckling mouse inoculation or organ homogenates and coculture of cell suspensions with Vero cells. By coculture techniques of whole blood, an otherwise undetectable viremia was demonstrated for both attenuated strains throughout the observation period. In contrast, XJ viremia was easily detected by direct techniques, as has already been shown. Attenuated virus was also shown to reach brain and bone marrow when coculture methods were employed. But titers were always markedly lower than those of the pathogenic strain. The sustained viremia demonstrated in guinea pigs infected with either attenuated strain explains the mode of viral dissemination and accounts for viral rescue and antigen detection from some organs. These results suggest that attenuated strains do not differ greatly in their invasive capacity in guinea pigs, but later on viral replication is impaired. Therefore, these findings reveal potential risks and should be noted when developing human vaccines. PMID- 2983014 TI - Rehospitalization in chronic schizophrenia. AB - This report on rehospitalization in chronic schizophrenia is based on a 1-year study of the postdischarge experiences of 119 chronic schizophrenics in New York City. The life table method of analysis identified the important role of discharge planning, community treatment compliance, and interpersonal stress in the patient's living environment in determining the number of days postdischarge that the patient remained in the community without further inpatient care. A mathematical model to predict days in hospital over the follow-up period, based on three specific components of time in hospital defined in numerical terms (PR, NR, LSR), was devised and tested. The first component, the experience of rehospitalization (PR), was determined by interpersonal stress, social supports, and aftercare treatment compliance. Adequacy of discharge planning, an intervention designed to link the patient to community treatment services, has its greatest impact in identifying number of rehospitalizations for the rehospitalized group (NR). Aftercare treatment compliance has its greatest effect in relation to length of subsequent rehospitalization episodes (LSR). Test of the model revealed that it can predict time in hospital within less than one half of a standard deviation of observed hospital days in approximately 50% of cases. PMID- 2983016 TI - The degradation of cytochrome c by hydrogen peroxide. AB - Cytochrome c is degraded by a large excess of hydrogen peroxide, leading to opening of the heme porphyrin ring and loss of the Soret absorption bands. The kinetic parameters of this reaction have been determined, and it is shown that a small concentration of oxygen is liberated at the same rate as degradation. Low level chemiluminescence and release of a hydroxylating species also accompany heme destruction. It is proposed that heme iron activates hydrogen peroxide to a more powerful oxidant, perhaps the hydroxyl radical, which remains bound to the heme iron and initiates attack on the porphyrin ring. Chemiluminescence appears to result from a side reaction involving singlet oxygen attack on the alpha methene bridge, yielding a dioxetane. The in vivo degradation of cytochrome c by excess hydrogen peroxide may interfere with respiration, accelerate aging, and enhance the metabolism of carcinogens. PMID- 2983015 TI - Formation of hydroxyl radicals from NADH and NADPH in the presence of copper salts. AB - The interaction of copper salts with NADH or NADPH in the presence of hydrogen peroxide at physiological pH is shown to produce hydroxyl radicals. The physiological significance of these results is discussed. PMID- 2983017 TI - Receptor-mediated phosphorylation of astroglial intermediate filament proteins in cultured astroglia. AB - Primary cultures of purified astroglia have been shown to exhibit a variety of membrane receptors that regulate intracellular cyclic AMP levels. The experiments described in this paper were completed to examine the effect of such receptor agonists on protein phosphorylation in intact astroglia. An analysis of 32P labelled proteins derived from whole cell extracts and separated via two dimensional gel electrophoresis indicated that increasing cyclic AMP levels in astroglia stimulated the phosphorylation of two distinct proteins that had apparent molecular weights/isoelectric points (pI) of 51K/6.0 and 57K/5.7. Similar experiments with cultured meningeal cells indicated that only the 57K/5.7 protein was phosphorylated in response to elevated levels of cyclic AMP. The 51K/6.0 protein was never observed in gels derived from meningeal cells. Immunoblot experiments indicated that the 51K/6.0 protein stained with antiserum to glial fibrillary acidic protein (GFAP) and the 57K/5.7 protein stained with antibodies to vimentin. Concentration-effect studies indicate that these proteins are maximally phosphorylated at concentrations of receptor agonists that only slightly elevate cyclic AMP levels. All receptor agonists that have been shown to increase cyclic AMP levels appear similarly efficacious with respect to increasing the phosphorylation of the two proteins. These experiments suggest that the membrane receptors present on astroglia function, in part, to regulate phosphorylation of the intermediate filament proteins GFAP and vimentin. PMID- 2983018 TI - Expression of A and B types of monoamine oxidase in differentiated neuroblastoma hybrid cells. AB - The total activities of monoamine oxidase (MAO) and the ratio of type B/type A activities were determined in mouse neuroblastoma N1E-115 cells, and in NX31T and NG108-15 hybrid cells derived from mouse neuroblastoma X rat sympathetic ganglion hybrid or mouse neuroblastoma X rat glioma hybrid cells. N1E-115 and NX31T cells possessed type A activities exclusively, although NG108-15 cells showed both type A (65-90%) and type B (10-35%) MAO activities. The activity of type A MAO in NX31T and N1E-115 cells was relatively constant during culturing periods in the presence or absence of dibutyryl cyclic AMP (Bt2cAMP), whereas total MAO activity and the ratio of type B MAO/type A MAO in NG108-15 cells increased as a function of culture periods. Prostaglandin E1 (PGE1) and theophylline, the best known combination to increase intracellular cyclic AMP content of NG108-15 cells, caused similar increases of MAO and of the type B/type A ratio in NG108-15 cells. The results suggest that MAO activity and expression of MAO B activity are regulated in NG108-15 cells in a cyclic AMP-dependent manner. PMID- 2983019 TI - Effects of systemically administered lithium on phosphoinositide metabolism in rat brain, kidney, and testis. AB - A single subcutaneous dose of 10 mEq/kg LiCl gives rise to an increase in the cerebral cortex level of myo-inositol-1-P (I1P) that closely follows cortical lithium levels and, at maximum, is 40-fold above the control value. Kidney and testis show smaller increases in I1P level following LiCl administration. The I1P level is still sixfold greater than that of untreated rat cortex 72 h later. In cortex, parallel increases also occur in myo-inositol-4-P (I4P) and myo-inositol 1,2-cyclic-P (cI1,2P), whereas myo-inositol-5-P (I5P) remains unchanged. The cortical increases in I1P and I4P levels are partially reversed by administering 150 mg/kg of atropine 22 h after the LiCl, treatment that does not affect cI1,2P. When doses of LiCl from 2 to 17 mEq/kg are given, the cerebral cortex levels of I1P and myo-inositol, measured 24 h later, are found to reach a plateau at about 9 mEq/kg of LiCl, whereas cortical lithium levels continued to increase with greater LiCl doses. Levels of all three of the brain phosphoinositides are unchanged by the 10 mEq/kg LiCl dose, as is the uptake of 32Pi into these lipids. Chronic dietary administration of LiCl for 22 days showed that the effects of lithium on I1P and myo-inositol levels persist for that period. Over the course of the chronic administration of the lithium, levels of I1P, myo-inositol, and of lithium in cortex remained significantly correlated. We believe that these increases in inositol phosphates result from endogenous phosphoinositide metabolism in cerebral cortex and that lithium is capable of modulating that metabolism by reducing cellular myo-inositol levels. The size of the effect is a function of both lithium dose and the degree of stimulation of receptor-linked phosphoinositide metabolism. This property of lithium may explain part of its ability to moderate the symptoms of mania. Our chronic study suggests that prolonged administration of LiCl does not result in compensatory changes in myo inositol-1-P synthase or myo-inositol-1-phosphatase. PMID- 2983021 TI - Influence of delta-opioid receptors on production of labeled methionine5 enkephalin in murine neuroblastoma cells. AB - Synthesis of methionine5-enkephalin by intact cells of murine neuroblastoma clone N1E-115 has been demonstrated both immunocytochemically and biochemically. In addition, N1E-115 cells possess homogeneous enkephalin (delta) receptors which inhibit prostaglandin E1-induced intracellular cyclic AMP formation. An assay was developed for measuring de novo synthesis of methionine5-enkephalin by pulsing cells in culture with radioactive methionine and isolating this pentapeptide to radiochemical purity by a procedure that included immunoaffinity chromatography specific for oxidized methionine5-enkephalin. This assay indicated that production of radiolabeled-methionine5-enkephalin was increased upon lengthy exposure of intact N1E-115 cells in the late logarithmic phase of growth to a nonproteolyzable analog of methionine5-enkephalin. This increase in synthesis of intracellular methionine5-enkephalin relative to control cells was prevented by prior incubation of the clone with naloxone, indicating that the response was mediated by the delta receptor. PMID- 2983020 TI - The picrotoxinin binding site and its relationship to the GABA receptor complex. AB - The binding characteristics of [3H] alpha-dihydropicrotoxinin to the picrotoxinin binding site were investigated in membrane preparations of adult rat forebrain and living cultures of rat cerebral cortex. The binding of [3H]alpha dihydropicrotoxinin to rat forebrain was decreased by lysing, treating with Triton X-100, and heating. Coincubation with gamma-aminobutyric acid (GABA), benzodiazepines, or alterations in the Na+ or Cl- composition of the media had no effect on the binding to the rat brain preparation. However, in the living neurons in tissue culture both GABA and diazepam significantly decreased the binding of [3H]alpha-dihydropicrotoxinin. The dose-response relationships for GABA antagonism of [3H]alpha-dihydropicrotoxinin binding and for picrotoxinin antagonism of the GABA enhancement of [3H]flunitrazepam binding in cultured cortical neurons were also investigated. The Hill coefficients for these actions were reciprocal, suggesting that they result from complementary interactions between the binding sites for GABA and picrotoxinin. These data support the association of the picrotoxinin binding site with the postsynaptic GABA receptor complex. PMID- 2983022 TI - Effect of electrical stimulation on phosphoinositide metabolism in rat sciatic nerve in vivo. AB - The metabolism of phosphoinositides in rat sciatic nerves in vivo during electrical stimulation was studied. Nerves were prelabeled by injection of [2-3H] myo-inositol alone for periods of 2 and 20 h or together with [32P]orthophosphate for 2 h and then electrically stimulated (100 Hz) for 5 or 20 min. Contralateral unstimulated nerve served as the control. When tritiated myo-inositol was used alone for prelabeling the nerves, approximately 6% and 14% of the label was incorporated into lipids after 2 h and 20 h, respectively. Both 5 and 20 min of electrical stimulation caused an insignificant change in the percentage of radioactivity recovered in lipids from the nerves prelabeled with either myo inositol or with a mixture of myo-inositol and phosphate. The proportion of label associated with phosphoinositides of nerves prelabeled with myo-inositol for both 2 h and 20 h showed an increase in phosphatidyl-inositol-4-phosphate at the expense of phosphatidylinositol in stimulated nerves. Similar results were obtained with nerves prelabeled for 2 h with a mixture of [32P]orthophosphate and [2-3H]myo-inositol. No significant changes in the radioactivity associated with water-soluble inositol phosphates were found in stimulated versus control nerves. PMID- 2983023 TI - Fluorescence polarization analysis, lipid composition, and Na+, K+-ATPase kinetics of synaptosomal membranes in feline GM1 and GM2 gangliosidosis. AB - Neurochemical studies were performed on synaptosomal membranes from cats with GM1 or GM2 gangliosidosis to examine possible mechanisms of neuronal dysfunction in these disorders. The basic hypothesis tested was that deficient ganglioside catabolism causes increased ganglioside content of synaptosomal plasma membrane which in turn disrupts normal function. Fluidity characteristics of synaptosomal membranes were examined using fluorescence polarization. Results showed markedly reduced membrane fluidity in both GM1 and GM2 gangliosidosis. These results were supported by a second study which revealed that isolated synaptosomal membranes of GM1 gangliosidosis cats had a 24-fold increase in total ganglioside content caused predominantly by excess GM1, a 2.3-fold increased cholesterol content, and a 1.4-fold increased phospholipid content. Finally, kinetic analysis of synaptosomal plasma membrane Na+,K+-ATPase from cats with GM1 gangliosidosis showed negligible differences in kinetic parameters compared with controls. Thus, the enzyme appeared protected from the global membrane changes in fluidity and composition. These observations provide evidence for a pathogenetic mechanism of neuronal dysfunction in the gangliosidoses while demonstrating protection of certain vital functional components, such as Na+,K+-ATPase. PMID- 2983024 TI - Comparison of proteins involved with cyclic AMP metabolism between synaptic membrane and postsynaptic density preparations isolated from canine cerebral cortex and cerebellum. AB - Synaptic membrane and postsynaptic density (PSD) fractions isolated from canine cerebral cortex and cerebellum were assayed for the following proteins: adenylate cyclase and phosphodiesterase (PDE) activities against cyclic AMP and cyclic GMP, the regulatory subunit of the cyclic AMP-dependent protein kinase, and the substrate proteins for this kinase. The results were expressed on the basis of both the protein content of the fractions and the number of synapses in the synaptic membrane fractions. The number of synapses on a constant protein content basis was about three times higher in the cerebral cortex synaptic membrane fraction than in the comparable cerebellar fraction. Adenylate cyclase activity was from 3.4 to 5.6 times higher in the cerebral cortex membrane fraction than in the cerebellar membrane fraction based on protein content but only slightly higher based on synapse counts. PSD fractions had no adenylate cyclase activity. The cyclic AMP-PDE activity was from 17 to 27 times higher in the cerebral cortex membrane fraction than in the cerebellar membrane fraction based on protein content, and about five times higher based on synapse counts. By doing PDE histochemistry at the electron microscopy level it was found that all the cerebral cortex PSDs in the isolated fraction contained PDE activity, none being found associated with the broken-up material in the fraction. The amount of the regulatory subunit of the cyclic AMP-dependent protein kinase was about equal in the two fractions based on protein, but about one-third lower in cerebral cortex fraction than in cerebellar fractions. In the cerebral cortex membrane fraction the primary substrate for the cyclic AMP-dependent protein kinase is synapsin I, with much lower amounts in the cerebellar membrane fraction. The PSD fraction from the two sources also showed these differences in synapsin I content. In the cerebellar membrane fraction, the primary substrate for the enzyme is a approximately 245,000 Mr protein not found in the cerebral cortex membrane fraction. The findings that the turnover of cyclic AMP is much higher in cerebral cortex synapses than in cerebellar synapses, and that differences are found between the cerebral cortex and cerebellum with regard to the substrate proteins for the cyclic AMP-dependent protein kinase indicate a divergence in the effect of cyclic AMP between cerebral cortex and cerebellar synapses. PMID- 2983026 TI - Avermectin B1a modulation of gamma-aminobutyric acid/benzodiazepine receptor binding in mammalian brain. AB - The anthelminthic natural product avermectin B1a (AVM) modulates the binding of gamma-aminobutyric acid (GABA) and benzodiazepine (BZ) receptor ligands to membrane homogenates of mammalian brain. The potent (EC50 = 40 nM) enhancement by AVM of [3H]diazepam binding to rat or bovine brain membranes resembled that of barbiturates and pyrazolopyridines in being inhibited (partially) by the convulsants picrotoxin, bicuculline, and strychnine, and by the anticonvulsants phenobarbital and chlormethiazole. The maximal effect of AVM was not increased by pentobarbital or etazolate. However, AVM affected BZ receptor subpopulations or conformational states in a manner different from pentobarbital. Further, unlike pentobarbital and etazolate, AVM did not inhibit allosterically the binding of the BZ receptor inverse agonist [3H]beta-carboline-3-carboxylate methyl ester, nor did it inhibit, but rather enhanced, the binding of the cage convulsant [35S]t-butyl bicyclophosphorothionate to picrotoxin receptor sites. AVM at submicromolar concentrations had the opposite effect of pentobarbital and etazolate on GABA receptor binding, decreasing by half the high-affinity binding of [3H]GABA and related agonist ligands, and increasing by over twofold the binding of the antagonist [3H]bicuculline methochloride, an effect that was potentiated by picrotoxin. AVM also reversed the enhancement of GABA agonists and inhibition of GABA antagonist binding by barbiturates and pyrazolopyridines. These overall effects of AVM are unique and require the presence of another separate drug receptor site on the GABA/BZ receptor complex. PMID- 2983025 TI - Transient states of adenylate cyclase in brain membranes. AB - Basal activity of adenylate cyclase from the amygdala of sheep brain and the neostriatum of turkey brain decays in two phases at 37 degrees C. The first phase is rapid (t1/2 = 2.3 +/- 0.3 min) and results in the loss of 60-70% of basal activity. The second phase is slow (t1/2 approximately 100 min) during which time the catalytic units denature irreversibly. The GTP analogue guanosine-5' (beta gamma imino) triphosphate (p[NH]ppG) prevents the rapid decay by stabilizing the enzyme at its initial level of activity and also reactivates the enzyme to initial levels during or immediately following the early phase, indicating that denaturation of neither the guanylnucleotide units nor the catalytic units causes the rapid decline in basal activity. Activation by p[NH]ppG is rapid at 37 degrees C, but the binding of p[NH]ppG to the guanylnucleotide subunit also occurs at nonactivatory temperatures. This is determined by the protection of catalytic units from thermal or N-ethylmaleimide inactivation after extensive washing. Thus, at 25 degrees C all of the catalytic units can be stabilized by saturating p[NH]ppG concentrations. At 0 degree C, 35% of the catalytic units can be stabilized by saturating p[NH]ppG concentrations within 30 s. The half saturation constant for the binding of p[NH]ppG at 0 degree C is identical to that derived in an assay at 37 degrees C, or after an incubation of the membranes for 10 min at 45 degrees C, when the process of thermal denaturation is 80% complete (K1/2 approximately 3 +/- 2 microM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983027 TI - Formation of free fatty acid and ceramide during brain handling: lability of sphingomyelin. AB - Intact brain and brain homogenates readily form free fatty acids and ceramides, even in the cold during subcellular isolation procedures. The fatty acid formation is slightly stimulated by chelators and might be due to phospholipid hydrolysis by lysosomal phospholipases. The ceramide formation is accompanied by loss of sphingomyelin and is apparently due to the action of neutral, metal ion activated sphingomyelinase. The latter reaction is inhibited by EDTA whereas both degradative processes are inhibited by mercuriphenylsulfonate, the thiol-reacting inhibitor. Cerebroside does not seem to be a source of accumulated ceramide. PMID- 2983028 TI - Modulation of [3H]diazepam binding in rat cortical membranes by GABAA agonists. AB - GABAA receptor agonists modulate [3H]diazepam binding in rat cortical membranes with different efficacies. At 23 degrees C, the relative potencies for enhancement of [3H]diazepam binding by agonists parallel their potencies in inhibiting [3H]gamma-aminobutyric acid [( 3H]GABA) binding. The agonist concentrations needed for enhancement of [3H]diazepam binding are up to 35 times higher than for [3H]GABA binding and correspond closely to the concentrations required for displacement of [3H]bicuculline methochloride (BMC) binding. The maximum enhancement of [3H]diazepam varied among agonists: muscimol = GABA greater than isoguvacine greater than 3-aminopropane sulphonic acid (3APS) = imidazoleacetic acid (IAA) greater than 4,5,6,7-tetrahydroisoxazolo (4,5,6) pyridin-3-ol (THIP) = taurine greater than piperidine 4-sulphonic acid (P4S). At 37 degrees C, the potencies of agonists remained unchanged, but isoguvacine, 3 APS, and THIP acquired efficacies similar to GABA, whereas IAA, taurine, and P4S maintained their partial agonist profiles. At both temperatures the agonist induced enhancement of [3H]diazepam binding was reversible by bicuculline methobromide and by the steroid GABA antagonist RU 5135. These results stress the importance of studying receptor-receptor interaction under near-physiological conditions and offer an in vitro assay that may predict the agonist status of putative GABA receptor ligands. PMID- 2983029 TI - Induction of myelin components: cyclic AMP increases the synthesis rate of 2',3' cyclic nucleotide 3'-phosphohydrolase in C6 glioma cells. AB - In an effort to determine the factors that stimulate myelin synthesis, we investigated the mechanism by which dibutyryl cyclic AMP induces the activity of the myelin enzyme, 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP; EC 3.1.4.37), in C6 glioma cells. Immunotitration experiments and measurements of the accumulation of [35S]methionine-labeled CNP showed that dibutyryl cyclic AMP increased the amount of CNP in the cells but not the catalytic activity per molecule of the enzyme. Moreover, inhibition of protein synthesis with cycloheximide abolished induction of enzyme activity. Dibutyryl cyclic AMP doubled the rate of CNP synthesis but had no effect on the half-life of the enzyme (approximately 33 h). The induction was partially blocked by the inhibitors of mRNA synthesis, cordycepin or alpha-amanitin. Thus, cyclic AMP induces the synthesis of CNP. PMID- 2983030 TI - Low activity of angiotensin-converting enzyme in cerebral microvessels of young spontaneously hypertensive rats. AB - Angiotensin-converting enzyme (ACE) activity was measured in microvessels prepared from cerebral cortices of 4-week-old spontaneously hypertensive rats (SHR). The Vmax value of the ACE activity in the cerebral microvessels of SHR was lower than that of Wistar Kyoto controls of the same age by 25% without difference in Km value for substrate. The low activity of ACE in the cerebral microvessels of young SHR indicates that in this animal model of hypertension the function of ACE is genetically altered in the cerebral microvessels, which may be correlated with the alteration of the cerebral microcirculation and pathogenesis of hypertension. PMID- 2983031 TI - Search for cytomegalovirus in postmortem brain tissue from patients with Huntington's chorea and other psychiatric disease by molecular hybridization using cloned DNA. AB - Postmortem human brain extracts were examined for the presence of human cytomegalovirus (CMV) DNA by molecular hybridization using a dot blot technique. The method was able to detect picogram quantities of homologous DNA, but CMV specific hybridization was detected in only one of 83 brains examined. The positive case came from a patient who had received immunosuppressive therapy. We were not able to confirm the report that CMV is present in the brains of patients with Huntington's chorea, nor was CMV detected in the temporal cortex of brains from schizophrenic patients. Our findings are discussed in relation to the methodology for investigating a possible viral etiology of some neuropsychiatric diseases. PMID- 2983032 TI - Long-term persistence of intrathecal virus-specific antibody responses after herpes simplex virus encephalitis. AB - Paired sera and cerebrospinal fluids (CSF) from nine surviving patients were collected 4.5 to 8 years after acute herpes simplex (HS) virus encephalitis. Oligoclonal bands of IgG were detected in the CSF of all, and seven patients had an elevated CSF IgG index. Antibodies to HS, varicella-zoster (VZ), measles, and cytomegalo viruses were analysed by enzyme-linked immunosorbent assay (ELISA) and by imprint immunofixation (IIF) of specimens separated by electrophoresis and by thin-layer electrofocusing. Intrathecal synthesis of HS and VZ IgG antibodies was demonstrated in all and of measles IgG antibodies in one patient by both methods. Intrathecal synthesis of HS IgA antibodies was demonstrated by ELISA in three and by IIF in seven patients; the latter method also disclosed intrathecal synthesis of VZ IgA antibodies in two. No patient had intrathecal synthesis of viral IgM antibodies. The intrathecally synthesized antibodies demonstrated by IIF displayed oligoclonal characteristics. The IIF analyses as well as virus absorption tests indicated that the intrathecally synthesized VZ IgG and IgA antibodies could be explained as HS antibodies cross-reacting with VZV. The results indicate that a long-term persistence of intrathecal antibody responses to HS virus is a common feature after acute HS encephalitis. The intrathecal production of measles IgG antibodies in one case may reflect a similar persistence of non-specific immune responses induced during the acute infection. PMID- 2983033 TI - Do true remissions in myasthenia really exist? An electrophysiological study. AB - To answer the question whether true remissions in myasthenia gravis occurred, 20 patients were studied with a history of evident, typical myasthenia but in full clinical remission. Two control groups served as a comparison: one of 10 healthy volunteers and the other of 10 patients with generalized, presently active myasthenia. In 17 of the 20 patients in remission single-fibre EMG (SFEMG) abnormalities were found, indicating some subclinical disturbances of neuromuscular transmission (in 3 cases the results were overtly pathological, in 11 cases moderately pathological, and in 3 cases slightly pathological). The abnormalities found in the remission group were much less distinct than those in the group with clinically symptomatic myasthenia. A case illustrates these findings: a man in full remission following thymoma extirpation in whom the SFEMG revealed subclinical disturbances of the neuromuscular transmission. Three weeks later a full clinical relapse occurred. The results obtained suggest that in the majority of patients the remissions are only apparently complete. A pathological jitter in a patient in remission calls for special attention: the patient must be spared any immunological stimuli such as vaccination, injections of sera, infections etc.; no drugs can be given which may increase the neuromuscular block; an immunosuppressive course of treatment should be considered; prognosis should be reconsidered. PMID- 2983034 TI - Phenytoin-induced orofacial dyskinesia. A case report. AB - A 56-year-old man with a small glioblastoma multiforme in the right parasagittal region developed orofacial dyskinesia and slight writhing movement of his hands during treatment with phenytoin and phenobarbitone. The serum concentration of phenytoin was within the therapeutic range. The involuntary movements subsided following the withdrawal of the drugs. Phenytoin-induced involuntary movements have not been described previously in a case with such a small parasaggital tumour treated with phenytoin at a serum concentration of therapeutic range. PMID- 2983035 TI - Neural circuits in the flight system of the locust. AB - Circuitry in the flight system of the locust, Locusta migratoria, was investigated by use of intracellular recording and staining techniques. Neuronal connections were established by recording simultaneously from neuropile segments of pairs of identified interneurons. Brief depolarizing current pulses delivered to interneurons 301 and 501 reset the flight rhythm in a phase-dependent manner, thus establishing the importance of these neurons in rhythm generation. Interneuron 301 was found to make a strong delayed excitatory connection with 501 and to receive a short-latency inhibitory connection from 501. The circuit formed by 301 and 501 appears suited for promoting rhythmicity in the flight system. The delayed excitatory potential recorded in 501 following each spike of 301 was reversed by hyperpolarizing 501. This potential and short-latency inhibitory postsynaptic potentials from 301 to other interneurons were blocked with the application of picrotoxin. We conclude that the delayed excitation is produced via a disynaptic pathway from 301 to 501, with 301 inhibiting in a graded manner the tonic release of transmitter from one or more unidentified intercalated neurons. Interconnections between the 301-501 circuit and other identified interneurons were discovered. This circuitry can account for two features of the flight motor pattern recorded in deafferented preparations. These features are the constant-latency relationship between depolarizations in elevator and depressor motoneurons and the relatively constant duration of depressor motoneuron bursts. The locust flight system shares general features with other described rhythm-generating systems. These include the occurrence of graded interactions, the probability of multiple oscillatory mechanisms, and a predominance of inhibitory connections. Its uniqueness lies in the way that components and processes are assembled and operate. PMID- 2983036 TI - Apparent movement of optic terminals out of a local postsynaptically blocked region in goldfish optic tectum. AB - In goldfish tectum, alpha-bungarotoxin (alpha BTX) blocks postsynaptically generated field potentials elicited by either photic or electrical stimulation, but leaves presynaptic activity unaffected. To assess the chronic effects of blocking transmission on synaptic stability, small restricted areas of synaptic block were created by slow, low-pressure microinjection of toxin-Ringer's solution from a micropipette. Local transmission was blocked, but field potentials outside the injected region were unchanged. Radioautography of 125I alpha BTX demonstrated that 6 days postinjection it was still highly localized and concentrated in discrete synaptic laminae of the tectal neuropil, in agreement with its known tight binding in goldfish brain (18). Electrophysiological maps made 6-8 days postinjection showed that few if any optic terminals could be recorded in the toxin-blocked zone, but there was no scotoma in the visual field. Terminals with receptive fields that would normally have corresponded to the region of injection were instead recorded from surrounding tectal areas. Since the normal units were also recorded there, this produced very large multiunit receptive fields. Apparently the optic terminals within the toxin-blocked area moved outward and innervated neighboring areas. In a minority of cases, no silent areas were noted and only the enlarged multiunit receptive fields were seen in the injection zone. All tecta injected with alpha BTX, but none injected with Ringer's, showed these disturbances in the map at the treated area at 6-8 days postinjection. By 2-3 wk, the maps in the alpha BTX injected tecta had returned to normal. There were no silent areas and no enlarged receptive fields. The movement of the optic arbors was shown to be dependent on activity in the optic fibers. In fish receiving intraocular injections of tetrodotoxin (TTX) for the first 4-6 days, no changes were seen in the retinotectal maps recorded after the effects of the TTX wore off. When the entire surface of the tectum was blocked with multiple injections of alpha BTX, normal maps were recorded at 6-8 days postinjection. Thus in order to produce movements in optic terminals, the fibers require activity and an opportunity to make effective synapses in a nearby zone. PMID- 2983037 TI - Modification of saccadic eye movements by GABA-related substances. I. Effect of muscimol and bicuculline in monkey superior colliculus. AB - Our previous observations led to the hypothesis that cells in the substantia nigra pars reticulata (SNr) tonically inhibit saccade-related cells in the intermediate layers of the superior colliculus (SC). Before saccades to visual or remembered targets, cells in SNr briefly reduce that inhibition, allowing a burst of spikes of SC cells that, in turn, leads to the initiation of a saccadic eye movement. Since this inhibition is likely to be mediated by gamma-aminobutyric acid (GABA), we tested this hypothesis by injecting a GABA agonist (muscimol) or a GABA antagonist (bicuculline) into the superior colliculus and measured the effects on saccadic eye movements made to visual or remembered targets. An injection of muscimol selectively suppressed saccades to the movement field of the cells near the injection site. The affected area expanded over time, thus suggesting the diffusion of muscimol in the SC; the area never included the other hemifield, suggesting that the diffusion was limited to one SC. One of the monkeys became unable to make any saccades to the affected area. Saccades to visual targets following injection of muscimol had longer latency and slightly shorter amplitudes that were corrected by subsequent saccades. The most striking change was a decrease in the peak velocity of the saccade, frequently to less than half the preinjection value. Saccades to remembered targets following injection of muscimol also showed an increase in latency and decrease in velocity, but in addition, showed a striking decrease in the accuracy of the saccades. The trajectories of saccades became distorted as if they were deflected away from the affected area. After muscimol injection, the area over which spontaneous eye movements were made shifted toward the side ipsilateral to the injection. Saccades toward the contralateral side were less frequent and slower. In nystagmus, which developed later, the slow phase was toward the contralateral side. In contrast to muscimol, injection of bicuculline facilitated the initiation of saccades. Injection was followed almost immediately by stereotyped and apparently irrepressible saccades made toward the center of the movement field of the SC cells at the injection site. The monkeys became unable to fixate during the tasks; the fixation was interrupted by saccadic jerks made to the affected area of the visual field and then back to the fixation point.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983038 TI - Modification of saccadic eye movements by GABA-related substances. II. Effects of muscimol in monkey substantia nigra pars reticulata. AB - The preceding study (21) showed that a gamma-aminobutyric acid (GABA) agonist or antagonist injected into the superior colliculus (SC) disrupted saccadic eye movements. The purpose of the present experiments was to determine whether this result was due to altering the inhibitory input to the SC from the substantia nigra pars reticulata (SNr). SNr cells are themselves inhibited by GABA. Injection of muscimol, a GABA agonist, into the SNr should increase the inhibition acting on SNr cells and should reduce the inhibition acting on the SC. If the effects of GABA inhibition in the SC results from terminals originating in the SNr, muscimol in the SNr should act like bicuculline in the SC. Muscimol in the SNr has the same general effect as bicuculline in the SC. The monkey made irrepressible saccades toward the contralateral visual field where cells in the SNr at the injection site had their visual or movement field. During visual fixation saccadic jerks occurred, interspersed with spontaneous saccades, instead of saccades to visual targets or to remembered targets. Saccades to remembered targets were more vulnerable to these saccadic intrusions than were saccades to visual targets. Since muscimol in the SNr acts like bicuculline in the SC, we conclude that a substantial fraction of GABA-mediated inhibitory inputs in the SC originates from the SNr. These experiments, in conjunction with previous experiments, show that the SNr exerts a tonic inhibition on saccade-related cells in SC and that this inhibition is mediated by GABA. The role of the SNr in initiation of saccades to remembered targets is particularly important since these saccades are more severely disrupted by muscimol in the SNr as well as in the SC. We suggest that both of these conclusions about eye movement might apply to skeletal movements as well. First, the basal ganglia contribute to the initiation of movement by a release of the target structure from tonic inhibition. Second, this mechanism is particularly critical of the movements based on stored or remembered signals that are not currently available as incoming sensory inputs. PMID- 2983039 TI - The effect of fibroblast growth factor on PC12 cells. AB - PC12 cells, which differentiate morphologically and biochemically into sympathetic neuron-like cells when treated with nerve growth factor, also respond to fibroblast growth factor. Some of the changes induced by fibroblast growth factor are similar to those seen after nerve growth factor treatment. Specifically, pituitary fibroblast growth factor causes the formation of processes initially comparable to those produced by nerve growth factor. However, in contrast to the outgrowth induced by nerve growth factor, which continues for several days, the outgrowth of processes induced by fibroblast growth factor ceases after about 3 days, even though fresh fibroblast growth factor is added. After about 6 days the processes induced by fibroblast growth factor have virtually disappeared. In this regard the processes induced by fibroblast growth factor are very similar to those induced by dibutyryl cyclic adenosine 3':5' monophosphate (dBcAMP). The addition of nerve growth factor and fibroblast growth factor together appears to produce a synergistic effect on process formation, as does the simultaneous addition of nerve growth factor and dBcAMP. Cells pretreated (or primed) with nerve growth factor are able to regenerate processes much more rapidly in the presence of nerve growth factor than cells which have not been pretreated. When fibroblast growth factor is added to cells primed with nerve growth factor, more rapid regeneration of processes also occurs. The regeneration of neurites in response to either factor is blocked by the addition of an inhibitor of methylation. The process formation induced by fibroblast growth factor is preceded, as is the outgrowth in response to nerve growth factor treatment, by an induction of ornithine decarboxylase, a decrease in the phosphorylation of a specific cytoplasmic protein, and an increase in the phosphorylation of a specific non-histone nuclear protein. The effects of fibroblast growth factor and of nerve growth factor on ornithine decarboxylase are additive. Fibroblast growth factor does not cause an increase in the activity of acetylcholinesterase; nerve growth factor does. Fibroblast growth factor does not appear to be acting through the nerve growth factor receptor. The binding of iodinated nerve growth factor to PC12 cells is specific and is not inhibited by the presence of fibroblast growth factor. In addition, anti-nerve growth factor serum does not interfere with the action of fibroblast growth factor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983040 TI - Norepinephrine and histamine potentiate the increases in cyclic adenosine 3':5' monophosphate elicited by vasoactive intestinal polypeptide in mouse cerebral cortical slices: mediation by alpha 1-adrenergic and H1-histaminergic receptors. AB - We have examined the interactions, in eliciting cAMP accumulation, between vasoactive intestinal polypeptide (VIP) and the three monoamines norepinephrine (NE), histamine (HIS), and serotonin (5-hydroxytryptamine, 5-HT) in mouse cerebral cortical slices. We have observed that NE and HIS, but not 5-HT, act synergistically with VIP to increase cAMP levels. The rank-order of potency of several adrenergic agonists in potentiating the effect of VIP on cAMP levels is the following: epinephrine greater than NE greater than phenylephrine much greater than clonidine, with EC50 of 2.2, 5, and 10 microM, respectively (clonidine being only marginally effective). This pharmacological profile is characteristic of the activation of alpha 1-adrenergic receptors. This contention is substantiated by the observation that the potentiating effect of NE is antagonized by the selective alpha 1-adrenergic antagonist prazosin at nanomolar concentrations. The potentiating effect of HIS is mediated by H1-histaminergic receptors since it is antagonized by the selective H1-receptor antagonist mepyramine but not by cimetidine, a selective H2-receptor antagonist. The synergistic interaction between VIP and NE is also observed in the presence of the adenosine antagonist theophylline, thus discarding the possibility of a mediation of the synergism by adenosine released by VIP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983041 TI - Neuropharmacological antagonism of the beta-carboline-induced "anxiety" response in rhesus monkeys. AB - A behavioral and physiological syndrome of stress-related responses was reported in primates following treatment with the benzodiazepine receptor antagonist, beta carboline-3-carboxylic acid ethyl ester (beta-CCE). The behavioral and physiological effects of beta-CCE are similar to those observed during stressful or "anxiety"-related conditions characterized in rhesus monkeys under natural conditions. Pharmacological agents which are known to antagonize anxiety responses in other paradigms were tested for their ability to antagonize the actions of beta-CCE. Diazepam (1 mg/kg) completely blocked the effects of beta CCE (200 micrograms/kg) on anxiety-related behaviors, heart rate and blood pressure, plasma catecholamines, cortisol, and adrenocorticotrophic hormone. A presynaptically active dose of the alpha-adrenoreceptor agonist, clonidine (10 micrograms/kg), significantly attenuated the effects of beta-CCE on all parameters, whereas the beta-adrenoreceptor agonist, propranolol (3 mg/kg), failed to alter the increases in plasma catecholamines, cortisol, or ACTH. In addition to these adrenergic agents, the serotonin antagonist, cyproheptadine (1 mg/kg), and the GABA-mimetic, 4,5,6,7-tetrahydroisoxazolo(5,4-C)pyrindin-3-ol (1 mg/kg), partially blocked the behavioral, physiological, and biochemical changes after beta-CCE. Manifestation of the complete "anxiety" syndrome evoked by beta CCE in primates may require the functional activity of several neurotransmitter systems. PMID- 2983042 TI - Thyrotropin-releasing hormone (TRH) receptors. Localization by light microscopic autoradiography in rat brain using [3H][3-Me-His2]TRH as the radioligand. AB - Thyrotropin releasing hormone (TRH) is a putative neurotransmitter in both the central and peripheral nervous system. In the present report, we have used autoradiography coupled with densitometric analysis of tritium-sensitive film to investigate the distribution of [3H][3-Me-His2]TRH [( 3H]MeTRH)-binding sizes in the rat brain. Previous pharmacological reports have established that many of these [3H]MeTRH-binding sites have a structure-activity profile consistent with being a physiological TRH receptor. A high level of TRH receptors were observed in the accessory olfactory bulb, lateral nucleus of the amygdala, dentate gyrus, and entorhinal cortex. Moderate levels of TRH receptors were observed in the rhinal cortex, hypothalamus, superior colliculus, several brainstem motor nuclei, and lamina I of the spinal trigeminal nucleus pars candalis, while low concentrations of receptors are present in the cerebral cortex, striatum and ventral horn of the spinal cord. Very low levels of receptors were observed in the globus pallidus and in most nuclei of the dorsal thalamus. Comparisons of the distribution of TRH receptors to TRH-immunoreactive content indicates that, while in some areas of the brain there is a rough correlation between levels of TRH peptide and its receptor, in most brain areas there is little obvious correlation between the two. While such a discrepancy has been observed for other peptides and their receptors, the extensive distribution of TRH receptors in the central nervous system does provide an explanation for the variety of behavioral effects observed when TRH is infused into the central nervous system. PMID- 2983043 TI - Differential ontogeny of multiple opioid receptors (mu, delta, and kappa). AB - We investigated the postnatal ontogeny of opioid receptors in rat brain under assay conditions which, when combined with computerized analysis, effectively reflect the developmental profile of high affinity binding to mu, delta, and kappa subpopulations. Concentrations of mu sites were assessed with the selective ligand 3H-[D-ala2,mePhe4,gly-ol5]enkephalin (DAGO). The other two sites were analyzed in binding assays with less selective radioligands but in the presence of specific unlabeled ligands which suppress cross-reactivity. We utilized 3H-[D ala2,D-leu5]enkephalin (DADL) in the presence of 10 nM DAGO to label delta sites and 3H-ethylketocyclazocine (EKC) in the presence of 100 nM DADL + 100 nM [D ala2,mePhe4,Met(0)ol5]enkephalin to detect kappa receptors. After birth, the density (femtomoles per milligram of wet weight) of mu sites declined for several days and then rose sharply over the next 2 weeks, increasing 2-fold by adulthood. Delta (delta) sites appeared in the second week postnatal and increased more than 8-fold in the next 2 weeks. Levels of kappa receptors were relatively low at birth and increased slowly (2-fold, overall). Computerized analyses of binding data revealed that DAGO and DADL were binding to single populations of sites throughout the postnatal period. DAGO and EKC affinities did not fluctuate in this period, whereas DADL affinities were low for the first week and then rose to adult levels. In summary, mu, kappa, and delta receptors exhibit differential postnatal developmental profiles. The former two are present at birth, whereas the latter appears in the second week. The postnatal increase for all three sites appear to be preceded by the previously demonstrated emergence of opioid peptides. PMID- 2983044 TI - Development of the rat superior cervical ganglion: ganglion cell maturation. AB - The development of superior cervical ganglion cells has been studied in the fetal rat. Sympathetic cells appear first in thoracic sites and, one day later, in cervical sites; localized proliferation among these cells gives rise to the superior cervical and stellate ganglia. The maturation of superior cervical ganglion cells was examined by staining these neurons with horseradish peroxidase in fetal preparations maintained in vitro. This method showed that cells begin to extend processes at widely different times, without regard to a given cell's position in the ganglion. Postganglionic axons appear as early as day 12 of gestation (E12), when only a small number of ganglion cells have emerged from the mitotic cycle. The axon generally originates from a point on the ganglion cell body that is oriented toward the route of subsequent axon extension. As the postganglionic axons grow, they do not branch within the superior cervical ganglion and branch only to a slight extent, if at all, within developing peripheral nerves. Axonal growth is rapid, and fibers reach relatively remote sympathetic targets as early as E15. Dendrites first appear on E14 and are elaborated by ganglion cells that have already extended their axons. By the end of gestation, the number of primary dendrites found on some cells already falls within the range found in maturity. PMID- 2983045 TI - Development of the rat superior cervical ganglion: ingrowth of preganglionic axons. AB - The growth of sympathetic preganglionic axons has been studied in the fetal rat. Preganglionic axons first emerge from the spinal cord and enter the paravertebral chain of ganglia between days 12 and 13 of gestation (E12 to E13). By E14, hundreds of preganglionic neurons have sent axons into the sympathetic chain. Even at this age, the arrangement of the preganglionic axons conforms to the adult organization in a number of ways. As in maturity, these axons leave the spinal cord only through the ventral root of the segment in which their cell bodies reside. Furthermore, axons grow into the superior cervical ganglion only from the same set of spinal segments as supply this ganglion in maturity, and in correct segmental proportions, although there is a slight rostral-to-caudal lag along the spinal cord in the full establishment of the projection. The development of appropriate projections in this system owes little to the growth of axons along stereotyped pathways. Rather, preganglionic organization apparently reflects a more general instruction that governs the relative number of axons from each spinal segment which course rostrally or caudally within the sympathetic chain. The appropriate distribution of preganglionic axons early in development suggests that a fiber's segmental level of origin biases the axon to grow in a particular direction. PMID- 2983046 TI - Development of the rat superior cervical ganglion: initial stages of synapse formation. AB - Synapse formation in the rat superior cervical ganglion has been investigated electrophysiologically and at the ultrastructural level. Preganglionic axons first enter the superior cervical ganglion between days 12 and 13 of gestation (E12 to E13), and on E13 a postganglionic response can be evoked by preganglionic stimulation. The susceptibility of this response to fatigue and to blocking agents indicates that it is mediated by cholinergic synapses. On E14, the overall strength of ganglionic innervation arising from different spinal segments already varies in a pattern resembling that found in maturity. However, the distribution of synapses on individual target cells gradually changes in the prenatal period. Transmission begins prior to the elaboration of ganglion cell dendrites; therefore, the first contacts to form are largely axosomatic. As dendrites appear (beginning on E14), ultrastructural evidence shows that synaptogenesis becomes focused upon these processes. The significance of these findings is discussed in relation to possible mechanisms for the formation of appropriate synaptic connections. PMID- 2983047 TI - Characterization of nitrobenzylthioinosine binding to nucleoside transport sites selective for adenosine in rat brain. AB - Nucleoside transport sites in rat brain membrane preparations were labeled with [3H]nitrobenzylthioinosine ([3H] NBI), a potent inhibitor of nucleoside transport systems. The membranes contained a single class of very high affinity binding sites with KD and Bmax values of 0.06 nM and 147 fmol/mg of protein, respectively. The displacement of [3H]NBI binding by various nucleosides, adenosine receptor agonists and antagonists, and known nucleoside transport inhibitors was examined. The Ki values (micromolar concentration) of [3H]NBI displacement by the nucleosides tested were: adenosine, 3.0; inosine, 160; thymidine, 240; uridine, 390; guanosine, 460; and cytidine, 1000. These nucleosides displayed parallel displacement curves indicating their interaction with a common site labeled by [3H]NBI. The nucleobases, hypoxanthine and adenine, exhibited Ki values of 220 and 3640 microM, respectively. Adenosine receptor agonists exhibited moderate affinities for the [3H]NBI site, whereas the adenosine receptor antagonists, caffeine, theophylline, and enprofylline, were ineffective displacers. The Ki values for cyclohexyladenosine, (+)- and (-) phenylisopropyladenosine, 2-chloroadenosine, and adenosine 5'-ethylcarboxamide were 0.8, 0.9, 2.6, 12, and 54 microM, respectively. These affinities and the rank order of potencies indicate that [3H]NBI does not label any known class of adenosine receptors (i.e., A1, A2, and P). The Ki values of other nucleoside transport inhibitors were: nitrobenzylthioguanosine, 0.05 nM; dipyridamole, 16 nM; papaverine, 3 microM; and 2'-deoxyadenosine, 22 microM. These results indicate that [3H]NBI binds to a nucleoside transporter in brain which specifically recognizes adenosine as its preferred endogenous substrate. This ligand may aid in the identification of CNS neural systems that selectively accumulate adenosine and thereby control "adenosinergic" function. PMID- 2983048 TI - Longitudinal topography and interdigitation of corticostriatal projections in the rhesus monkey. AB - Anterograde tracing methods were used to examine the topographic organization and interrelationship of projections to the neostriatum arising from various areas of association cortex. In contrast to the currently accepted topographic schema, all cortical areas examined project to longitudinal territories that occupy restricted medial-lateral domains of the neostriatum. The posterior parietal and superior arcuate cortices project to dorsolateral portions of the neostriatum; the dorsolateral and dorsomedial frontal cortices project centrally; and the orbitofrontal, anterior cingulate, and superior temporal projections are distributed to ventromedial regions of the caudate nucleus and putamen. In coronal section, cortical terminal fields form a diagonal strip, extending from the dorsal, ventricular border of the caudate nucleus, through the fiber bundles of the internal capsule, to the ventral margin of the putamen. Double labeling studies, in which two cortical areas were injected in the same animal, indicated that convergence of input within neostriatal domains is not governed by reciprocity of corticocortical connectivity. Thus, the interrelationship of projections arising from connectionally linked cortical areas ranged from nearly complete segregation of terminal fields (e.g., from dorsolateral prefrontal and orbital cortices) to extensive overlap of terminal domains (e.g., from frontal and temporal cortices). In the latter case, detailed analysis revealed that frontal and temporal terminals actually were interdigitated rather than intermixed within the zone of overlap. The present findings suggest a new conceptualization of corticostriatal topography in the primate which emphasizes the longitudinal arrangement of cortical terminal domains. Additionally, these findings provide a map for functional parcellation of the neostriatum on the basis of its cortical innervation which may prove useful to understanding normal striatal function, as well as the symptomatology associated with neostriatal injury and disease. PMID- 2983050 TI - Calf enlargement from S-1 radiculopathy. Report of two cases. AB - Two patients with painless hypertrophy of one calf had clinical and myelographic features compatible with an S-1 radiculopathy. Electromyographic studies revealed intermittent spontaneous discharges (bizarre repetitive potentials) in the affected extremity. Several patients have previously been described with S-1 radiculopathy and calf hypertrophy with or without spontaneous electrical activity. The hypertrophy in the two present cases could have been caused by continuous stretching (both patients were very active), by the abnormal electrical activity, or by a combination of the two. A neurogenic lesion must be excluded in patients with isolated hypertrophy of a calf muscle, even in the absence of radicular pain. PMID- 2983049 TI - Retinal projections and functional architecture of cortical areas 17 and 18 in the tyrosinase-negative albino cat. AB - The visual field representation and functional architecture of cortical areas 17 and 18 in albino cats were studied. In the same animals the distributions of ipsilaterally and contralaterally projecting retinal ganglion cells were determined by injecting horseradish peroxidase into the dorsal lateral geniculate nucleus or optic tract. All cats were tyrosinase-negative albinos (cc), not deaf white cats (W). The proportion of ipsilaterally projecting ganglion cells in the temporal retina of the albino cat was found to be much smaller than in the normal cat or in the Siamese cat. In the albino cat less than 5% of ganglion cells in temporal retina project ipsilaterally. Recordings from areas 17 and 18 provided evidence of a substantial representation of the ipsilateral hemifield in albino visual cortex; cells representing the contralateral and ipsilateral hemifields were often segregated into alternating zones in area 17 and were always segregated in area 18. Cells recorded at the borders of zones representing the ipsilateral and contralateral hemifields often had abnormal properties. Some border cells had two receptive fields separated by as much as 60 degrees of azimuth; one field subserved the contralateral hemifield (contralateral nasal retina) and the other subserved the mirror-symmetric part of ipsilateral hemifield (contralateral temporal retina). Receptive fields of cells subserving the two hemifields did not differ in size. The preferred orientations, preferred velocities, and other characteristics of the two fields were approximately the same; preferred orientation changed gradually and systematically across the borders of zones representing the two hemifields. Our results indicate that afferents representing nasal and temporal regions of retina of the same eye can segregate and form "hemiretina" domains in albino visual cortex. These afferents can also converge upon individual cortical cells in a fashion reminiscent of convergence of afferents from the two eyes upon binocular cells in the normal cortex. The organization of albino visual cortex is therefore different from the organization of Siamese visual cortex. This may be because, in the albino cat but not the Siamese cat, nearly all cells in temporal retina project contralaterally; afferents representing contralateral temporal retina are not at a significant competitive disadvantage in the albino. PMID- 2983052 TI - Complications of chymopapain chemonucleolysis. PMID- 2983053 TI - Imaging of the hepatocellular carcinoma using dynamic positron emission tomography with nitrogen-13 ammonia. AB - Dynamic PET using [13N]ammonia was performed in patients with primary hepatocellular carcinoma (hepatoma). All the tumors started to show remarkable accumulation of radioactivity from a very early period (within 150 sec after the radionuclide injection), whereas the radionuclide was more gradually accumulated in the liver. Central necroses of the tumors were visualized as low radioactivity areas. Daughter nodules of less than 2 cm were also visualized. This dynamic PET study is a valuable technique for the detection of the hepatoma. PMID- 2983051 TI - Neurological manifestations of the acquired immunodeficiency syndrome (AIDS): experience at UCSF and review of the literature. AB - In this review of the acquired immunodeficiency syndrome (AIDS), the authors have evaluated a total of 352 homosexual patients with AIDS or generalized lymphadenopathy managed at the University of California, San Francisco (UCSF), between 1979 and 1984. Of an initial unselected group of 318 patients, 124 (39%) were neurologically symptomatic, and one-third already had their neurological complaints at the time of presentation. An additional 210 AIDS patients with neurological symptoms have been reported in the literature. Thus, a total of 366 neurologically symptomatic patients with AIDS or lymphadenopathy are reviewed. Central nervous system (CNS) complications, encountered in 315 patients, included the following viral syndromes: subacute encephalitis (54), atypical aseptic meningitis (21), herpes simplex encephalitis (nine), progressive multifocal leukoencephalopathy (six), viral myelitis (three), and varicella-zoster encephalitis (one). Non-viral infections were caused by Toxoplasma gondii (103), Cryptococcus neoformans (41), Candida albicans (six), Mycobacteria (six), Treponema pallidum (two), coccidioidomycosis (one), Mycobacterium tuberculosis (one), Aspergillus fumigatus (one), and Escherichia coli (one). Neoplasms included primary CNS lymphoma (15), systemic lymphoma with CNS involvement (12), and metastatic Kaposi's sarcoma (three). Cerebrovascular complications were seen in four patients with hemorrhage and five with infarction. Five patients in the UCSF series had multiple intracranial pathologies, including two cases of simultaneous Toxoplasma gondii infections and primary CNS lymphoma, two cases of coexistent Toxoplasma gondii and viral infections, and one case of combined Toxoplasma gondii and atypical mycobacterial infection. Cranial or peripheral nerve complications, seen in 51 patients, included cranial nerve syndromes secondary to chronic inflammatory polyneuropathy (five), lymphoma (five), and Bell's palsy (five). Peripheral nerve syndromes included chronic inflammatory polyneuropathy (12), distal symmetrical neuropathy (13), herpes zoster radiculitis (six), persistent myalgias (two), myopathy (two), and polymyositis (one). In light of the protean behavior of AIDS and the problems related to the clinical, radiological, and serological diagnosis of the unusual and varied associated nervous system diseases, patients with AIDS and neurological complaints require a rigorous and detailed evaluation. The authors' experience suggests that biopsy of all CNS space-occupying lesions should be performed for tissue diagnosis prior to the institution of other therapies. PMID- 2983055 TI - Stimulated salivary clearance of technetium-99m pertechnetate. AB - A method to determine stimulated salivary clearance of pertechnetate is presented. It is easy to perform and separates normal patients (range 15.0 to 40.3 ml/min) from patients with known salivary disorders (range 1.2 to 10.6 ml/min). PMID- 2983054 TI - Radionuclide turnover studies on ectopic thyroid glands--case report and survey of the literature. AB - A 72-yr-old woman who had previously undergone partial thyroidectomy for "toxic goiter" and subhyoid gland resection presented with signs and symptoms of ectopic lingual thyroid at the base of the tongue. The combination of lingual subhyoid and pretracheal thyroid is a rarity with only three cases reported. Discovery of ectopic thyroid tissue should raise suspicion of other ectopic thyroid tissues along the path of embryologic migration from its origin to the porta hepatis. This may necessitate assessment of radionuclide uptake and imaging of numerous areas. Sodium levothyroxine is the mainstay of therapy. In patients with obstructive symptoms, 131I ablation of the ectopic thyroid tissues has proven successful and may be more advantageous than surgery. In addition, radioiodide studies during ablation therapy gave new information that the ectopic lingual thyroid had more rapid iodide turnover compared with the postsurgical pretracheal thyroid gland. PMID- 2983056 TI - Benign fibrous histiocytoma of the oral mucosa. PMID- 2983057 TI - Sex cord-stromal tumours of the ovary. AB - The aim of this review is to give a reasonably concise resume of our knowledge of the sex cord-stromal tumours of the ovary. Lipoid cell tumours of the ovary are often included within this broad category but this poorly defined and heterogenous group of neoplasms will not be considered here. This review is a selective one and no attempt is made to cover all aspects of sex cord-stromal tumours or to provide a complete bibliography. The histological features of many of the neoplasms in this group, particularly those which have been recently defined, are discussed but a consideration of differential histological diagnosis is excluded. The ultrastructural characteristics of the various neoplasms are considered only in terms of their relevance to histogenesis or metabolic activity. PMID- 2983058 TI - Viremia in hospitalized children with enterovirus infections. AB - In a prospective study during the summer and fall of 1982, enterovirus was isolated from 48 hospitalized children; in 29 (60%) enterovirus was isolated from CSF or blood, and in 19 (40%) only a presumptive diagnosis was established. Blood was positive in 21 (44%) and was the only positive specimen in two children. A presumptive diagnosis was provided within 4 days of admission in 38 (80%) and within 48 hours in 19 (40%) of the children from whom enterovirus was isolated. Viremia was most often detected in febrile infants younger than 3 months of age with a clinical picture simulating bacterial sepsis. The presence of viremia was inversely related to the presence of CSF pleocytosis and to virus isolation from CSF. The diagnosis of diseases caused by enterovirus is more accurate when blood culture is added to CSF stool and throat cultures. PMID- 2983059 TI - Long-term evolution of glycerol intolerance syndrome. PMID- 2983060 TI - Linitis plastica in a child. AB - Anaplastic infiltrative adenocarcinoma of the linitis plastica morphologic type is rare in children. The case of a 15-year-old child with this unusual form of gastric carcinoma is presented. One year postoperatively following radiation and chemotherapy, the child is free of disease. PMID- 2983061 TI - Further evidence that tetracyclines inhibit collagenase activity in human crevicular fluid and from other mammalian sources. PMID- 2983062 TI - Effects of the cannabinoids on physical properties of brain membranes and phospholipid vesicles: fluorescence studies. AB - The effects of four cannabinoids on the physical properties of brain synaptic plasma membranes (SPM), lipid extracts of SPM and phospholipid vesicles were evaluated using fluorescence probes. In vitro, the psychoactive cannabinoids, delta 9-tetrahydrocannabinol (delta 9-THC) and 11-hydroxyl-delta 9 tetrahydrocannabinol (11-OH-delta 9-THC) at concentrations of 1 and 3 microM decreased polarization of the fluorescence emission of 1,6-diphenyl-1,3,5 hexatriene (DPH) in SPM. At the same concentrations, cannabidiol (CBD) and cannabinol, cannabinoids devoid of marijuana-like psychoactivity, had no effect on DPH polarization. The effects of 11-OH-delta 9-THC and CBD on vesicles made from lipids extracted from SPM were identical to their effects on intact SPM. These changes in DPH polarization were not due to changes in fluorescence lifetime and indicate that, at low concentrations, the psychoactive cannabinoids increase the rotational mobility of DPH in the membrane core. In contrast, in SPM extracted lipids, both 11-OH-delta 9-THC and CBD decreased the mobility of stearic acid with an anthroyloxy label at both the second (2-AS) and twelfth (12 AS) carbon atoms. Studies of DPH polarization in various phosphatidylcholines (PC) demonstrated that the actions of the cannabinoids were dependent on initial bilayer fluidity. 11-OH-delta 9-THC was less effective at decreasing polarization of trimethylammonium DPH (TMA-DPH), a probe of the bilayer surface, than of DPH whereas CBD affected mobility of the two probes equally. Neither CBD nor 11-OH delta 9-THC altered DPH mobility in phosphatidylethanolamine, phosphatidylserine vesicles. These findings indicate that the psychoactive cannabinoids increase fluidity in the hydrophobic core of brain membranes and support a membrane perturbant hypothesis of the mechanism of delta 9-THC action. PMID- 2983063 TI - A1 and A2 adenosine receptor activation inhibits and stimulates renin secretion of rat renal cortical slices. AB - Previous studies by others have demonstrated that exogenous adenosine inhibits renin secretion in vivo. In the present experiments, we studied the effects of three adenosine receptor agonists [N6-cyclohexyladenosine (CHA), 2 chloroadenosine (2-CIA) and 5'-N-ethylcarboxamideadenosine (NECA)] on renin secretion of rat renal cortical slices. The effects were biphasic; submicromolar concentrations inhibited secretion concentration-dependently and the order of potency was CHA greater than 2-CIA greater than NECA. Micromolar and higher concentrations stimulated secretion concentration-dependently and the order of potency was reversed: NECA greater than 2-CIA greater than CHA. These results are consistent with the hypothesis that activation of A1 and A2 adenosine receptors produces inhibition and stimulation of secretion, respectively. Theophylline antagonized both the inhibitory effect of low concentrations of CHA and the stimulatory effect of higher concentrations, providing additional evidence for mediation by activation of cell-surface adenosine receptors. Calcium chelation abolished the inhibitory effect of CHA, suggesting that increased intracellular calcium mediates the inhibitory effect; on the other hand, the inhibitory effect was unaffected by membrane depolarization and calcium channel blockade, suggesting that CHA-induced inhibition is not due to calcium influx through voltage-sensitive calcium channels. Ouabain, vanadate and K-depolarization, all of which are believed to increase intracellular calcium, antagonized CHA- and NECA-stimulated renin secretion, suggesting that the stimulatory effect of these agonists is mediated by decreased intracellular calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983065 TI - Apparent kinetics of angiotensin converting enzyme: hydrolysis of [3H]benzoyl phenylalanyl-alanyl-proline in the isolated perfused lung. AB - Isolated perfused rabbit lungs were used to study the hydrolysis of [3H]benzoyl phenylalanyl-alanyl-proline [( 3H]BPAP), a synthetic substrate for angiotensin converting enzyme (ACE). Lungs were perfused, at constant flow rates, with physiologic medium containing added BPAP and the concentration of its metabolite, [3H]benzoyl-phenylalaline, was measured in lung effluent. Hydrolysis of BPAP (4.2 microM) was 64.1 +/- 3.3% at 37 degrees C and was significantly decreased (P less than .01) to 10.1 +/- 8.7% by the addition of the ACE inhibitor, MK422 (10(-6) M). Disappearance (i.e., hydrolysis) of immunoreactive angiotensin I was also inhibited by MK422. Hydrolysis of BPAP was saturable and calculated apparent kinetic constants were Km = 13 microM and Vmax = 50 nmol/sec/lung. When the perfusion medium temperature was 10 degrees C, apparent Km was unchanged, whereas Vmax was significantly (P less than .05) decreased. At BPAP concentrations sufficient to depress metabolism to less than 20%, perfusion pressure was unchanged. Hydrolysis of BPAP under first-order conditions was independent of flow over the range 10 to 50 ml/min. However, increase in flow rate to 100 ml/min/lung was associated with decreased BPAP metabolism. These data indicate that BPAP is a substrate for pulmonary ACE in vitro and substantiate its use in intact animals because: 1) it is without physiologic effect in high doses; and 2) calculated apparent kinetics in isolated lungs under these conditions of steady state concentrations were similar to those obtained from earlier studies that utilized bolus injection techniques in intact animals. PMID- 2983064 TI - Role of cyclic AMP protein kinase in decreased arterial cyclic AMP responsiveness in hypertension. AB - A decreased relaxation responsiveness to isoproterenol and forskolin is manifest in aortic smooth muscle isolated from spontaneously hypertensive rats (SHR) when compared with normotensive Wistar Kyoto (WKY) rats. Inasmuch as the effector of cyclic AMP (cAMP) is cAMP-dependent protein kinase, we sought to determine if alterations in this enzyme might be responsible for this decreased responsiveness to cAMP-increasing vasodilators. The concentration of cAMP protein kinase activity in aortic, carotid and caudal arteries (approximately 300 pmol/mg of protein per min) was similar in both WKY and SHR. Activity in femoral arteries from SHR and WKY rats was greater (approximately 600 pmol/mg/min); branches of the femoral artery from SHR had less protein kinase activity (660 pmol/mg/min) than their WKY counterparts (1000 pmol/mg/min). There were no differences between WKY and SHR in isozymic distribution of soluble cAMP protein kinase in any of these sources of arterial smooth muscle. Concentration and temporal-related relaxation of KCl-contracted aortic muscle strips by forskolin was associated with concomitant activation of cAMP protein kinase in both groups. The rate and extent of kinase activation was similar for both groups even though the rate and extent of relaxation was markedly less in SHR. These findings show that neither the concentration, isozymic distribution nor activation of cAMP-dependent protein kinase are different in aortic smooth muscle isolated from SHR when compared with WKY animals. Thus, decreased relaxation responsiveness to cAMP-increasing vasodilators is probably not related to events proximal to and including activation of arterial cAMP-dependent protein kinase. PMID- 2983066 TI - Enhanced in vivo responsiveness of presynaptic angiotensin II receptor-mediated facilitation of vascular adrenergic neurotransmission in spontaneously hypertensive rats. AB - Presynaptic angiotensin II (AII) receptor-mediated facilitation of vascular adrenergic neurotransmission was studied in the in situ, blood-perfused mesentery of 13- to 16-week-old spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar Kyoto rats (WKY). Mesenteric arterial perfusion pressure frequency-response curves to periarterial adrenergic nerve stimulation (PNS) and dose-response curves to exogenous norepinephrine (NE) were obtained in SHR and WKY. The effects of the following treatments on the mesenteric vascular perfusion pressure responses (PPR) to PNS and NE were studied: All alone infused i.a. at 1 and 5 ng/min; All infused at 5 ng/min after [Sar1-lle8]All infused at 20 ng/min; [Sar1-lle8]All infused at 20 ng/min alone; captopril alone at 0.1 mg/kg i.v.; All infused i.a. at 0.3 and 1 ng/min after captopril at 0.1 mg/kg and angiotensin I injected at 3 dose levels after captopril at 0.1 mg/kg. Control PPR to PNS and NE were greater in SHR than in WKY. Comparisons of PPR in SHR to those in WKY were made, therefore, at the predetermined PPR levels of 15, 20, 30, 40, 50, 60 and 70 mm Hg. All alone shifted the PNS frequency-response curve to the left to a greater extent in the SHR than in the WKY when infused at 5 ng/min but not when infused at 1 ng/min. Both infusion rates of All had significantly different effects on the dose-response curves to NE in WKY and SHR. The effects of All infusion (5 ng/min) on both the response to PNS and to NE were antagonized completely by the concurrent infusion of [Sar1-lle8] All at 20 ng/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983067 TI - Pulmonary metabolism of exogenous enkephalins in isolated perfused rat lungs. AB - Metabolism of enkephalins during transit through the pulmonary circulation may be of significance in regulating systemic levels of these opioids. To determine whether Leu- and Met-enkephalin are metabolized by the pulmonary circulation, [3H]Tyr-Leu-enkephalin (10 microM) or [3H]Tyr-Met-enkephalin (10 microM) were each administered to isolated rat lungs perfused in a recirculating manner with a physiologic salt solution and a recently developed high-performance liquid radiochromatographic analytical method was used to identify and quantitate metabolites in the perfusion medium. Both Leu- and Met-enkephalin were metabolized in a curvilinear, time-dependent manner. The principal metabolites were identified as tyrosine and Tyr-Gly-Gly. Neither Tyr-Gly nor Tyr-Gly-Gly-Phe were detected in significant amounts. After a 20-min perfusion, residual Leu- or Met-enkephalin accounted for 28.4 and 21.5%, respectively, of the radioactivity present in the perfusate. In addition, 97% of the initial radioactivity for both Leu- and Met-enkephalin were found in the perfusion medium, indicating that neither the parent compounds nor metabolites were avidly sequestered in pulmonary tissue. The angiotensin converting enzyme inhibitor, captopril (18 microM) blocked the formation of Tyr-Gly-Gly and attenuated slightly the production of tyrosine. Inhibition of aminopeptidase with bestatin (116 microM) blocked the formation of tyrosine and enhanced production of Tyr-Gly-Gly. Inhibition of enkephalinase with thiorphan (0.3 microM) did not appear to affect Met-enkephalin metabolism. These observations indicate that in isolated, buffer perfused rat lungs Leu- and Met-enkephalin are metabolized during pulmonary transit by at least two enzymes, angiotensin converting enzyme and aminopeptidase. PMID- 2983068 TI - Selective blockade of endothelium-dependent and glyceryl trinitrate-induced relaxation by hemoglobin and by methylene blue in the rabbit aorta. AB - Hemoglobin at 1 microM reduced and at 10 microM abolished the endothelium dependent relaxation induced by acetylcholine or by A23187 in rabbit aortic rings. Similarly, methylene blue at 10 microM reduced and at 50 microM abolished relaxation induced by acetylcholine and by A23187. Furthermore, hemoglobin (1-10 microM) and methylene blue (10-50 microM) each induced a dose-dependent inhibition of the endothelium-independent relaxation produced by glyceryl trinitrate, but neither had any effect on the relaxation produced by isoproterenol. The inhibitory effects of hemoglobin and methylene blue may be due to blockade of guanylate cyclase, as the rises in cyclic GMP content which accompany relaxation induced by acetylcholine, A23187 or glyceryl trinitrate were abolished. Isoproterenol-induced relaxation took place with no change in cyclic GMP content. Hemoglobin and methylene blue appear therefore to inhibit selectively vaso-relaxation induced by agents which increase cyclic GMP levels. Hemoglobin and methylene blue augment tone in aortic rings, particularly when endothelial cells are present, suggesting that the endothelium-derived relaxing factor (EDRF) might be released spontaneously in low concentrations. The possibility that hemoglobin inhibits endothelium-dependent and glyceryl trinitrate-induced relaxation by binding EDRF and nitric oxide, respectively, is discussed together with the proposal that methylene blue might produce its effects by oxidizing a component of guanylate cyclase, possibly a ferrous heme group linked to the enzyme molecule. Methylene blue might, in addition, interact directly with EDRF. PMID- 2983069 TI - Characterization of cholecystokinin receptor sites in guinea-pig cortical membranes using [125I]Bolton Hunter-cholecystokinin octapeptide. AB - [125I]Bolton Hunter-cholecystokinin octapeptide (BH-CCK8) has been prepared using a modified method and was used to study putative cholecystokinin (CCK) receptor sites in the guinea-pig cerebral cortex. Specific binding of [125I]BH-CCK8, defined as the difference in binding in the absence and presence of 10(-6) M CCK8, was 70% of total binding. In saturation experiments, the apparent dissociation constant (Kd) was 1 nM and total binding capacity was 28 fmol/mg of protein. In association experiments, conducted at 30 degrees C, binding of [125I]BH-CCIK8 reached equilibrium in approximately 150 min. Binding was stable for 4 hr and was reversed by the addition of unlabeled CCK8-sulfated. Dissociation of bound ligand was biphasic and the apparent T1/2 was 45 min. Analyses of kinetic experiments yielded an association rate constant of 0.58 X 10(8) min-1 M-1 and a dissociation rate constant for the slower component of 0.012 min-1. Dithiothreitol increased and N-ethylmaleimide decreased specific binding of [125I]BH-CCK8, indicating that CCK receptor sites involve sulfhydryl groups. In competition experiments, the potency of CCK4 was enhanced 50-fold with addition of protease inhibitors. The rank order of CCK-related peptides was CCK8 sulfated greater than or equal to Gastrin 17 greater than or equal to CCK33 greater than CCK4 greater than or equal to CCK8-desulfated. Proglumide, a proposed CCK antagonist in the periphery and brain, was inactive at 10(-3) M. The specificity of [125I]BH-CCK8 binding sites are similar to that reported for [125I]BH-CCK33. PMID- 2983071 TI - Dissociation between the excitatory and "excitotoxic" effects of quinolinic acid analogs on the striatal cholinergic interneuron. AB - Analogs of quinolinic acid were tested for excitatory properties in evoking neurotransmitter release from striatal cholinergic interneurons and for their ability to lesion these same neurons in vivo (excitotoxin activity). The ability of these analogs to inhibit the specific binding of several ligands thought to label excitatory amino acid receptors was also investigated. Dipicolinic acid (2,6-pyridine dicarboxylic acid) was found to be as potent and as efficacious as quinolinic acid (2,3-pyridine dicarboxylic acid) at N-methyl-D-aspartate (NMDA) type receptors mediating [3H]acetylcholine release from striatal slices. However, unlike quinolinate, the structure of NMDA is not superimposable upon that of dipicolinic acid. Moreover, unlike quinolinic acid, dipicolinic acid injected intrastriatally did not produce detectable excitotoxic lesions. Most unexpectedly, phthalic acid (1,2-benzene dicarboxylic acid), which lacks a nitrogen, also evoked [3H]acetylcholine release from striatal slices, apparently by acting at NMDA-type receptors. Phthalic acid was equipotent to quinolinic acid and dipicolinic acid but possessed less intrinsic activity than these compounds in evoking [3H]acetylcholine release. Despite its lack of a nitrogen and low intrinsic activity, intrastriatal injection of phthalic acid produced axon sparing lesions of intrinsic cell bodies, like quinolinic acid as assessed by neurochemical and histologic methods. Quinolinic acid, dipicolinic acid and phthalic acid were moderately potent inhibitors (Kl = approximately equal to 100 microM) of the specific binding of 2-[3H]amino-7-phosphonoheptanoic acid a compound thought to be a competitive antagonist of NMDA-type receptors. In contrast, these three compounds failed to inhibit the chloride-dependent or chloride-independent binding of L-[3H]glutamate or [3H]kainic acid. The present results suggest a major dissociation between the structure-activity relationships for "excitotoxicity" vis-a-vis excitation as reflected in the [3H]acetylcholine release model, for compounds acting at NMDA-type receptors. PMID- 2983072 TI - New hydroxyapatite ceramic materials: potential use for bone induction and alveolar ridge augmentation. PMID- 2983070 TI - Tolerance and cross-tolerance to the antisecretory effects of enkephalins on the guinea-pig ileal mucosa. AB - The antisecretory properties of opiates in the guinea-pig ileum have been shown previously to be mediated through interactions with delta-like opiate receptors present in the intestinal mucosa. The present investigation examined the development of tolerance to opioid-induced alterations in intestinal ion transport processes. Osmotic minipumps continuously delivering the prototypic delta-opioid agonist [D-Ala2,D-Leu5]enkephalin (DADLE) (5 micrograms/hr) or the potent mu-opiate agonist fentanyl (10 micrograms/hr) over a 5-day period were implanted s.c. into guinea pigs; control animals did not receive chronic drug infusions. DADLE, DADLE ethylamide and [D-Ala2,D-Met5]enkephalin dose-dependently decreased base-line transepithelial potential difference and short-circuit current in isolated segments of ileal mucosa from untreated control animals, with an order of potency of DADLE greater than [D-Ala2, D-Met5 enkephalin greater than DADLE ethylamide. In tissues from DADLE-infused animals, the antisecretory dose effect curves of the three enkephalin analogs displayed downward shifts to the right compared to control conditions. In contrast, the potency of DADLE was significantly increased in tissues from animals chronically infused with and rendered physically dependent upon fentanyl. The administration of the opioid antagonists naloxone, diprenorphine or the selective delta-opioid antagonist M 154, 129 produced no significant changes in short-circuit current of mucosal segments from either DADLE- or fentanyl-infused animals. Chronic administration of either DADLE or fentanyl did not significantly alter the effects of nonopioids, bombesin, somatostatin or epinephrine, on ion transport; however, the efficacy but not the potency of neurotensin in increasing short-circuit current was attenuated after chronic DADLE infusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983073 TI - Virus infections in immunocompromised patients: their importance and their management. AB - Opportunistic viral infections were investigated in 156 adult patients admitted over one year to a medical oncology service: 35% of the total group and 65% of those with acute leukaemia experienced viral infections, 79% of which were with viruses of the herpes group. Surprisingly few enteric viruses were recovered. Reactivation of herpes simplex virus in the brains of these immunosuppressed patients was suggested by the demonstration by nucleic acid hybridization of herpes simplex virus DNA sequences in neurones and endothelial cells in patients with evidence of past infection with virus. Acyclovir was effective in therapy and prophylaxis. Twenty-three strains from 7 patients were tested for sensitivity to this antiviral: in 3 instances clinical resistance was observed but the strains were fully sensitive in vitro, as were all other strains tested. PMID- 2983075 TI - Angiotensin converting enzyme inhibitors. (Mercaptoaroyl)amino acids. AB - A series of (mercaptoaroyl)amino acids and related compounds was synthesized and tested for ability to inhibit angiotensin converting enzyme (ACE). The most active compound was N-(3-chloro-2-mercaptobenzoyl)-N-cyclopentylglycine, having an in vitro I50 = 0.28 microM. Substitution of the aromatic 3-position by small polar groups enhanced ACE inhibitory activity, whereas bulky groups diminished it. Alteration of the beta relationship between the mercaptan and amide carbonyl or masking of the thiol by acylation reduced activity. Replacement of the thiol by nitro, hydroxy, or carboxy gave compounds lacking ACE inhibitory activity. PMID- 2983074 TI - Hormonal aspects of prostatic cancer: a review. PMID- 2983076 TI - Conformational analysis and active site modelling of angiotensin-converting enzyme inhibitors. AB - The discovery of captopril as a potent, orally active inhibitor of angiotensin converting enzyme (ACE) led to the recent development of many series of novel structures with similar biological activity. To date, however, all of these inhibitors are flexible or semiflexible molecules, and there is therefore no clear definition of the conformational requirements for ACE inhibition. In an effort to solve this problem, we have carried out conformational energy calculations on a series of eight structurally diverse ACE inhibitors. Comparison of the low-energy conformations available to these molecules leads to the conclusion that there is a common low-energy conformation throughout the series. The calculations thus define the structural and conformational requirements for ACE inhibition. Expansion of this model to the receptor level has been achieved by considering possible alternative receptor sites for each of the molecules in its proposed biologically active conformation and leads to an active-site model for ACE which may be useful for the design of further inhibitors. PMID- 2983077 TI - Growth cones of developing retinal cells in vivo, on culture surfaces, and in collagen matrices. AB - The outgrowth of axons from the early retina in vivo is compared with that from retinal explants in two types of culture systems. The normal time course of axonal growth along the primordial optic pathway to the optic tectum is characterized, using tritiated proline and horseradish peroxidase (HRP) as anterograde tracers. The rate of axonal elongation in vivo is estimated to be about 32 micron/hr at 22 degrees C. The HRP technique allows visualization of retinal growth cones in vivo. Observations can thus be made on their microanatomy and on the environment through which they navigate. The growth cones of retinal ganglion cells in the embryo have lamellipodia and fairly short filopodia (approximately 10 micron) which are directed forward. The growth cones are found near the pial surface of the brain but do not seem to maintain contact with it. Two culture systems were developed to investigate axonal pathways in vitro. In the first, different substrates and culture media were explored. Results indicate that growth cones prefer a polyornithine substrate over a collagen one. The media that promotes the best neurite outgrowth consists of L15 (60%), fetal calf serum (10%) and Xenopus embryo extract (1 mg/ml). Time-lapse video monitoring of substrate cultures reveals an average rate of outgrowth of about 18 micron/hr with great variability. The growth cones in these cultures are large, flattened, and complex compared to those in vivo, and their filopodia extend in many different directions. The second culture system is a collagen gel infiltrated with growth medium. In these conditions neurite outgrowth more closely mimics that in vivo. The rate is faster than on substrates, and the growth cones appear morphologically similar to those in the embryo. Preliminary experiments using the gel culture system to test for chemotaxis of retinal axons toward their targets failed to demonstrate such an effect. PMID- 2983078 TI - Growth cone-target interactions in the frog retinotectal pathway. AB - The growth cones of retinal ganglion cell axons were studied in the optic tract and tectum with horseradish peroxidase (HRP) histochemistry and electron microscopy. The ganglion cell growth cones has many morphological features similar to those described in vitro and in other in vivo systems. However, we found that some processes formed highly differentiated terminal arborizations, while retaining growth cones on many of their branches. In addition, ultrastructural examination of the tectal neuropil revealed that many ganglion cell axonal processes had characteristics of both growth cones and presynaptic endings. These findings are discussed in the context of the hypothesis of shifting connections and the evidence that retinotectal map formation involves several mechanisms, including a process that depends on the action potential activity in the optic fibers. PMID- 2983079 TI - Differential expression of surface proteins, virus receptors and histocompatibility antigens in SV40-transformed human choroid cells and their clones. AB - Human choroid (HC) cells transformed by Simian virus 40 (HC/SV40) and some of their soft agar-derived clones were utilized to study the influence of SV40 transformation on the expression of some differentiation parameters. The presence of receptors for DNA and RNA viruses, known to induce cytopathic effects in target cells, always occurred in normal HF and HC/SV40 cells as well as in their derived clones, whereas differences were observed in the expression of influenza A and Sindbis virus receptors. Immunofluorescence assays showed an increase in the expression of class I (HLA-A/B/C) and class II (DR) histocompatibility antigens in hyperdiploid clones compared with both the hypodiploid clones and the normal human cell counterpart. The two surface glycoproteins, fibronectin and laminin, appear only partially correlated with the transformed phenotype of the cells both at optical (immunofluorescence) and supraoptical (immunoelectron microscopy) level. This study confirms the SV40 transformation induces in human choroid cells a modulation of the considered differentiation parameters and that this may be due to a different insertion mode of the SV40 genome in HC cells. PMID- 2983080 TI - Simultaneous presentation of a small-cell carcinoma involving the ovary and the uterine endometrium. AB - A case of simultaneous presentation of a small-cell carcinoma involving the ovary and the uterine endometrium is reported. We consider the endometrium as the primary localization of the tumor. The epithelial origin and neuroendocrine differentiation were confirmed by electron microscopy. Tumor cells were attached by small desmosomes, and in the cytoplasm typically neurosecretory granules measuring 100-200 nm were found. Immunohistochemically, no content of polypeptide hormones (ACTH, Calcitonin, Gastrin, Glucagon, Insulin, Somatostatin and VIP) were encountered. The tumor stained strongly for neuronspecific enolase. The histogenetic possibilities are shortly presented. PMID- 2983081 TI - Purification of simian virus 40 large T antigen by immunoaffinity chromatography. AB - Simian virus 40 large T antigen from lytically infected cells has been purified to near homogeneity by immunochromatography of the cell extract on a protein A Sepharose-monoclonal antibody column. The resulting T antigen retains biochemical activity; i.e., it hydrolyzes ATP and binds to simian virus 40 DNA at the origin of replication. PMID- 2983082 TI - The circular intracellular form of Epstein-Barr virus DNA is amplified by the virus-associated DNA polymerase. AB - Selective DNA extraction and hybridization procedures were used to estimate the relative number of covalently closed circular viral genomes in cultures of Epstein-Barr virus (EBV)-transformed cells. In virus-producing P3HR-1 cultures that were exposed for 11 days to phosphonoacetic acid or to acyclovir, the content of covalently closed circular EBV DNA was reduced ca. 70% relative to a control culture without drug. The EBV plasmid content of Raji, a virus nonproducer cell line, was not reduced by exposure to these compounds. When P3HR 1 cultures were exposed to 12-O-tetradecanoylphorbol-13-acetate, the number of circular genomes per cell increased. These findings indicate that two enzyme activities synthesize circular EBV DNA and that the virus-associated DNA polymerase synthesizes most of the circular EBV DNA in a virus producer culture. It is suggested that the circular genomes synthesized by the viral enzyme are intermediates in the syntheses of linear virus DNA. PMID- 2983083 TI - Analysis of the major transcripts encoded by the long repeat of human cytomegalovirus strain AD169. AB - In this report, we describe the size and kinetics of appearance of RNAs from the long repeat of human cytomegalovirus. The most abundant RNA from this region was a 2.7-kilobase (kb) species that was detected throughout the infection and was most abundant at 27 and 72 h after infection. The 2.7-kb RNA was the only major species detected with a probe that included the terminus of the long repeat and the heterogeneous L-S junction region. Other transcripts were detected with probes from the internal portion of the long repeat, including an immediate-early RNA of 1.3 kb, early and late RNAs of 1.2 kb, and minor late transcripts of 4.4, 3.6, 3.3, and 1.8 kb. S1 nuclease and exonuclease VII protection analyses of RNA from immediate-early, early, midpoint, and late times in the infection indicated that the major 2.7-kb RNA was not spliced and that the RNA mapped within the long repeat, 1.6 kb from the heterogeneous region. No evidence for temporally regulated changes in transcription initiation, splicing, or choice of 3' end of this RNA was observed. Nuclease protection analysis also demonstrated that the second most abundant late RNA from this region, the 1.2-kb species, was not spliced and had the same polarity as the 2.7-kb RNA. The 1.2-kb also mapped entirely within the long repeat, with its 3' terminus 1.7 kb upstream from the 5' terminus of the 2.7-kb RNA. PMID- 2983084 TI - Recombination between a defective retrovirus and homologous sequences in host DNA: reversion by patch repair. AB - The genomes of mammalian species contain multiple copies of sequences homologous to exogenous retroviruses. When a mutant retrovirus carrying a lethal deletion in an essential viral gene was introduced into mammalian cells, revertant viruses appeared and spread throughout the culture. Analysis of one such revertant showed that the mutation had been repaired by homologous recombination with endogenous sequences. Our results suggest that defective retroviruses can draw upon the genetic complement of the host cell to repair lesions in viral genes. PMID- 2983085 TI - Fine structure of the cottontail rabbit papillomavirus mRNAs expressed in the transplantable VX2 carcinoma. AB - We analyzed the polyadenylated mRNAs transcribed from the cottontail rabbit papillomavirus genomes present in the domestic rabbit transplantable carcinoma line VX2, employing a combination of S1 nuclease mapping and primer extension techniques with vector M13-based single-stranded DNA probes. Each of the two major mRNA species (1,860 and 1,110 bases long) contained two exons which corresponded to the E6-E7 and E2-E4 open reading frames. The splice donor site for these transcripts was located at position 1371 at the beginning of the E1 open reading frame. Consequently, the splicing event did not lead to the fusion of the E region proximal (E6 and E7) and distal (E2, E4, or E5) open reading frames. The translation of the polycistronic RNA could result in the production of E6 and E7 proteins alone or of an additional E1-E4 fusion product if translation reinitiation can occur after the E7 stop codon. A heterogeneity was detected in the 5' ends of the longer transcript; E6 transcripts could thus yield a full-length or a truncated E6 protein. We also detected a minor subset of mRNAs covering the E2-E4 coding region and including at least three species with estimated sizes of 4.2, 2.8, and 1.8 kilobases. All the viral transcripts detected in the VX2 tumor cells were polyadenylated at the same site (position 4367) 20 base pairs beyond the first AATAAA signal which borders the E region. PMID- 2983086 TI - Evidence for a direct role for both the 175,000- and 110,000-molecular-weight immediate-early proteins of herpes simplex virus in the transactivation of delayed-early promoters. AB - We reconstructed the regulated induction of delayed-early (DE) transcription that occurs during herpes simplex virus (HSV) infection by using a transient expression system in which recombinant target genes were cotransfected into Vero cells together with intact activating genes. Plasmids containing cloned HSV-1 or HSV-2 immediate-early (IE) genes stimulated by up to 100-fold the expression from recombinant constructs containing the bacterial chloramphenicol acetyltransferase (CAT) gene under the control of the DE promoter/regulatory region from the genes for an HSV-2 38,000-molecular-weight (38K) protein and the HSV-1 thymidine kinase. This activation was specific to hybrid genes containing DE regulatory regions since no significant increases in expression were observed in cotransfection experiments with the CAT gene without any promoter region or under the control of a number of other regulatory regions, including an HSV-1 IE regulatory region, the complete or enhancerless early regulatory region of simian virus 40, and an inducible cellular promoter/regulatory region. By using a variety of cotransfected plasmids containing individual or different combinations of HSV-1 or HSV-2 IE genes, we show that of the five known IE genes, two, those coding for the 175K and 110K polypeptides, each possessed the ability to stimulate expression from both DE promoters. Cleavage of the input plasmids within the known coding regions for the 175K and 110K proteins abolished stimulation of DE/CAT gene expression, whereas cleavage outside the coding regions had no effect on stimulation. We conclude that stimulation of CAT expression occurred exclusively by a transactivation mechanism in which the products encoded by these IE genes acted on the DE hybrid constructs at the transcription level. No transcriptional stimulatory function was demonstrated for the IE 68K and 63K proteins, although our results indicate that the IE 12K protein may augment the DE stimulatory activity of the 175K and 110K proteins. PMID- 2983087 TI - Structural analysis of the varicella-zoster virus gp98-gp62 complex: posttranslational addition of N-linked and O-linked oligosaccharide moieties. AB - Varicella-zoster virus specifies the formation of several glycoproteins, including the preponderant gp98-gp62 glycoprotein complex in the outer membranes of virus-infected cells. These viral glycoproteins are recognized and precipitated by a previously described monoclonal antibody designated monoclone 3B3. When an immunoblot analysis was performed, only gp98 was reactive with monoclone 3B3 antibody; likewise, titration in the presence of increased concentrations of sodium dodecyl sulfate during antigen-antibody incubations caused selective precipitation of gp98 but not gp62. Further structural analyses of gp98 were performed by using the glycosidases endo-beta-N acetylglucosaminidase H (endoglycosidase H) and neuraminidase and two inhibitors of glycosylation (tunicamycin and monensin). In addition to gp98, antibody 3B3 reacted with several intermediate products, including gp90, gp88, gp81, and a nonglycosylated polypeptide, p73. Since gp98 was completely resistant to digestion with endoglycosidase H, it contained only complex carbohydrate moieties; conversely, gp81 contained mainly high-mannose residues. Polypeptide p73 was immunodetected in the presence of tunicamycin and designated as a nascent recipient of N-linked sugars, whereas gp88 was considered to contain O-linked oligosaccharides because its synthesis was not affected by tunicamycin. The ionophore monensin inhibited production of mature gp98, but other intermediate forms, including gp90, were detected. Since the latter product was similar in molecular weight to the desialated form of gp98, one effect of monensin treatment of varicella-zoster virus-infected cells was to block the addition of N acetylneuraminic acid. Monensin also blocked insertion of gp98 into the plasma membrane and, as determined by electron microscopy, inhibited envelopment of the nucleocapsid and its transport within the cytoplasm. On the basis of this study, we reached the following conclusions: the primary antibody 3B3-binding epitope is located on gp98, gp98 is a mature product of viral glycoprotein processing, gp98 contains both N-linked and O-linked oligosaccharide side chains, gp90 is the desialated penultimate form of gp98, gp88 is an O-linked intermediate of gp98, gp81 is the high-mannose intermediate of gp98, and p73 is the unglycosylated precursor of gp98. PMID- 2983089 TI - Hypothermia-inducing peptide promotes recovery of vesicular stomatitis virus from persistent animal infections. AB - A single injection of the hypothermia-inducing neuropeptide bombesin resulted in an excellent recovery system for reisolating viruses from Swiss albino mice infected with vesicular stomatitis virus even up to 90 days after infection. The virus was recovered from a cell homogenate prepared from whole brain tissue 24 h after intracerebral injection of bombesin; brain cells were cocultivated with BHK 21 cell monolayers and then plaqued on BHK-21 cells at 31 degrees C. All of the recovered viruses were identified as vesicular stomatitis virus by antibody neutralization and peptide analyses of some of the structural proteins. However, some of the recovered viruses were altered with regard to tryptic peptide maps, temperature sensitivity, and central nervous system disease induced compared with the viruses used to initiate the infection. Most of the recovered viruses induced a similar disease when reinoculated intracerebrally into mice, characterized by hind-leg paralysis 4 to 6 days after infection. Two of the recovered viruses were lethal, however, resulting in a relatively rapid generalized wasting disease and death in 3 to 4 days. PMID- 2983088 TI - Inhibition of cellular DNA polymerase alpha and human cytomegalovirus-induced DNA polymerase by the triphosphates of 9-(2-hydroxyethoxymethyl)guanine and 9-(1,3 dihydroxy-2-propoxymethyl)guanine. AB - The triphosphates of 9-(2-hydroxyethoxymethyl)guanine and 9-(1,3-dihydroxy-2 propoxymethyl)guanine were examined for their inhibitory effect on highly purified cellular DNA polymerase alpha and human cytomegalovirus (Towne strain) induced DNA polymerase. These two nucleoside triphosphates competitively inhibited the incorporation of dGMP into DNA catalyzed by the DNA polymerases. The virus-induced DNA polymerase had greater binding affinity for the triphosphate of 9-(2-hydroxyethoxymethyl)guanine (Ki, 8 nM) than for the triphosphate of 9-(1,3-dihydroxy-2-propoxymethyl)guanine (Ki, 22 nM), although the nucleoside of the latter compound was strikingly more effective against human cytomegalovirus replication in cell cultures than the nucleoside of the former. The Ki values of these two nucleoside triphosphates for alpha polymerase were 96 and 146 nM, respectively, and were 7- to 12-fold higher than those for the virus induced enzyme. These data indicated that virus-induced DNA polymerase was more sensitive to inhibition by these two nucleoside triphosphates than was the cellular alpha enzyme. PMID- 2983090 TI - In vitro phenotypic markers of a poliovirus recombinant constructed from infectious cDNA clones of the neurovirulent Mahoney strain and the attenuated Sabin 1 strain. AB - Infectious cDNA corresponding to the entire genome of the attenuated Sabin strain of type 1 poliovirus has been inserted into EcoRI site of bacterial plasmid pBR325. Two consecutive PstI fragments (nucleotide positions 1814 to 3421) of the infectious cDNA of the Sabin 1 strain were replaced by the corresponding DNA fragments prepared from an infectious DNA clone of the genome of the virulent Mahoney strain of poliovirus type 1. The exchanged segment encodes capsid protein VP1 and part of capsid protein VP3, a region in which a large number of amino acid differences between the attenuated Sabin and the parental, neurovirulent Mahoney strain cluster. The recombinant virus was obtained by DNA transfection of HeLa S3 cells, and several in vitro phenotypes of the virus were compared with those of the parental viruses. The recombinant virus was recognized by a neutralizing monoclonal antibody specific to the Mahoney strain. Growth of the Sabin strain of poliovirus has been shown to be quite dependent upon the bicarbonate concentration (d marker). The growth of the recombinant virus, however, was not highly dependent upon the concentration of bicarbonate in cell culture media, and thus resembled that of the Mahoney strain. On the other hand, the temperature-sensitive multiplication (rct marker) and the small-plaque morphology of the recombinant virus corresponded to the phenotype of the Sabin 1 strain. The in vitro recombination of infectious cDNA clones of genomic RNA and subsequent analysis of the growth properties of the recombinant virus have allowed us to correlate specific mutations in the genome of an RNA virus with certain biological characteristics of that virus. PMID- 2983091 TI - Qualitative and quantitative analyses of Epstein-Barr virus early antigen diffuse component by western blotting enzyme-linked immunosorbent assay with a monoclonal antibody. AB - We report the use of monoclonal antibody against the early antigen diffuse component (anti-EA-D) of Epstein-Barr virus (EBV) to analyze, both qualitatively and quantitatively, the expression of EA-D in various human lymphoblastoid cell lines activated by chemical inducers. The kinetics of synthesis of EA-D in P3HR 1, B95-8, and Ramos/AW cells were similar in that they all reached the peak of synthesis on day 5 after induction. Surprisingly, no expression of EA-D was found in induced BJAB/GC, an EBV-genome-containing cell line. EBV-negative cell lines, BJAB and Ramos, were negative for EA-D. Raji cells had no detectable EA-D but responded rapidly to induction, reaching a peak on day 3. Superinfection of Raji cells also resulted in marked induction of EA-D, which reached a plateau between 8 to 12 h postinfection. Western blotting coupled with the enzyme-linked immunosorbent assay was employed to identify polypeptides representing EA-D. A family of four polypeptides with molecular weights of 46,000 (46K protein), 49,000, 52,000, and 55,000 were identified to be reactive with monoclonal anti-EA D antiserum. The pattern of EA-D polypeptides expressed in each cell line was different. Of particular interest was the expression of a large quantity of 46K protein both in induced Raji and P3HR-1 cells, but not in superinfected Raji cells. A 49K doublet was expressed in activated p3HR-1, B95-8, and Ramos/AW cells and in superinfected Raji cells. In addition, two distinct 52K and 55K polypeptides were expressed in induced Ramos/AW and superinfected Raji cells. However, none of these EA-D polypeptides was detectable in BJAB/GC, BJAB, Ramos, and mock-infected Raji cells. To approximate relative concentrations of EA-D in cell extracts, we employed the enzyme-linked immunosorbent assay and immunoblot dot methods by using one of the purified EA-D components to construct a standard curve. Depending upon the cell lines, it was estimated that ca. 1 to 3% (determined by the enzyme-linked immunosorbent assay) and 0.8 to 1.6% (determined by immunoblot dot) of total proteins from maximally induced cells were EA-D. These results suggest that differential expression of EA-D polypeptides could be of importance in the diagnosis of state of EBV infection. PMID- 2983092 TI - Characterization of a new simian virus 40 mutant, tsA3900, isolated from deletion mutant tsA1499. AB - The simian virus 40 (SV40) mutant tsA1499 contains an 81-base-pair deletion in the region of A gene encoding the C-terminal portion of the large T antigen. This mutant is particularly interesting, since it is a temperature-sensitive mutant that is apparently able to separate the lytic growth and transforming functions of the SV40 large T antigen at 38.5 degrees C. We report the isolation of a tsA1499 revertant (tsA1499-Rev) which is no longer temperature sensitive for lytic growth but still contains the 81-base-pair deletion of tsA1499. Marker rescue experiments with tsA1499-Rev or wild-type strain 830 (wt830) DNAs revealed that the original tsA1499 mutant contained a second mutation within the HindIII Fnu4HI restriction fragment between 0.425 and 0.484 map units. Sequencing of this DNA fragment from the tsA1499, tsA1499-Rev, and wt830 viruses revealed that tsA1499 contained a single-base transversion (C to G) at 0.455 map units (nucleotide 4261). This transversion resulted in the creation of a new RsaI cleavage site in the tsA1499 DNA and predicts an arginine-to-threonine substitution at amino acid position 186 in the mutant large T antigen. The DNA sequence of the tsA1499-Rev HindIII-Fnu4HI fragment was identical to that of wt830. To determine whether tsA1499 was temperature sensitive for lytic growth solely as a result of the newly discovered point mutation or because of a combination of the point and deletion mutations, a series of viruses were constructed which contained the point mutation, the deletion mutation, both mutations, or neither. This was done by ligating the PstI A and B DNA fragments from either tsA1499 or wt830 and transfecting the ligated DNA into BSC-1H monkey kidney cells. This experiment revealed that all viruses containing the point mutation (the tsA1499 PstI A DNA fragment) were temperature sensitive for lytic growth, regardless of the presence of the 81-base-pair deletion (the tsA1499 PstI B DNA fragment). This newly discovered point mutation, at nucleotide 4261, is therefore unique, since to our knowledge it is the first tsA mutation to be described in the 0.455-map-unit region of the SV40 genome. We then tested the effect of this unique mutation on the ability of the SV40 virus to transform cultured rat cells to anchorage independence.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983093 TI - Isolation from cats of an endogenous type C virus with a novel envelope glycoprotein. AB - A search for variant endogenous cat viruses led to a novel isolate. Although the major envelope glycoprotein of this virus was similar in size to that of an RD 114-like virus that was coisolated, it was unrelated to RD-114 or feline leukemia virus by immunological and biological criteria. This degree of dissimilarity suggests a different evolutionary progenitor from that for the RD-114 and feline leukemia virus viral envelopes. The novel virus did, however, code for gag gene polypeptides which are closely related to RD-114 virus. Neither the novel isolate nor the RD-114-like coisolate induced foci in S+L- cat cells which restrict focus induction by RD-114 virus. This suggests that the two viruses share a common genomic target of restriction which resides outside of the env region. PMID- 2983094 TI - Three intergenic regions of coronavirus mouse hepatitis virus strain A59 genome RNA contain a common nucleotide sequence that is homologous to the 3' end of the viral mRNA leader sequence. AB - cDNA clones that represent various portions of the coronavirus mouse hepatitis virus strain A59 genome RNA have been constructed. cDNAs were synthesized by transcription of genome RNA by using either oligo(dT) or random oligomers of calf thymus DNA as primers. These cDNAs were converted into double-stranded DNA and cloned into pBR322 by standard techniques. The resulting cloned viral DNA fragments were mapped to viral genes by hybridization with Northern blots of intracellular RNA from mouse hepatitis virus strain A59-infected cells. These cDNA clones map in six of the seven viral genes. Clone g344, 1.8 kilobases, is the largest and encompasses gene 5 (which encodes a nonstructural protein) and gene 6 (which encodes the E1 viral glycoprotein) as well as the intergenic regions preceding genes 5, 6, and 7. Sequencing of parts of this cloned DNA show that these three intergenic regions contain a common 11-nucleotide sequence. This sequence shares homology with the 3' end of the viral mRNA leader sequence. Thus, this common intergenic sequence may contain a binding site for a leader RNA that hybridizes to negative-strand viral RNA at the beginning of each gene to prime mRNA synthesis. The different degrees of homology between the leader and its putative binding site may influence the differential rates of transcription of the various viral mRNAs. PMID- 2983095 TI - Isolation of polyomavirus mutants multiadapted to murine embryonal carcinoma cells. AB - Polyomavirus mutants were isolated from PCC4 embryonal carcinoma cells infected with a variant strain of polyomavirus (ev 1001h) and were found to contain a tandem duplication overlapping the enhancers and the origin of replication. These mutants were able to infect several lines of embryonal carcinoma cells, including PCC4, F9, and LT1. The sequence and structure of one of these mutants are presented and compared with those of other PyEC PCC4 mutants previously described. PMID- 2983096 TI - Properties of the deoxycholate-solubilized HeLa cell plasma membrane receptor for binding group B coxsackieviruses. AB - Physical and chemical properties of deoxycholate-solubilized HeLa cell plasma membrane receptors for binding group B coxsackieviruses were determined. Receptors eluted from Sepharose 4B with an apparent molecular weight of 275,000 and sedimented with an S value of between 14.7 and 4.9 and a buoyant density of 1.06 to 1.10 g/cm3. Virus-binding activity was destroyed after treatment with proteases, glycosidases, and periodate but was unaffected by lipases or reducing or alkylating agents. Additionally, lectins, including concanavalin A, adsorbed receptors and inhibited virus attachment. The composite data suggested that glycoprotein is an integral part of the receptors for binding virus. PMID- 2983097 TI - Nucleotide sequence of avian sarcoma virus UR2 and comparison of its transforming gene with other members of the tyrosine protein kinase oncogene family. AB - The genome of avian sarcoma virus UR2 was completely sequenced and found to have a size of 3,165 nucleotides. The UR2-specific transforming sequence, ros, with a length of 1,273 nucleotides, is inserted between the truncated gag gene coding for p19 and the env gene coding for gp37 of the UR2AV helper virus. The deduced amino acid sequence for the UR2 transforming protein P68 gives a molecular weight of 61,113 and shows that it is closely related to the oncogene family coding for tyrosine protein kinases. P68 contains two distinctive hydrophobic regions that are absent in other tyrosine kinases, and it has unique amino acid changes and insertions within the conserved domain of the kinases. These characteristics may modulate the activity and target specificity of P68. PMID- 2983098 TI - Sequence of hepatitis B virus DNA incorporated into the genome of a human hepatoma cell line. AB - Seven copies of integrated hepatitis B virus (HBV) DNA and contiguous genomic DNA from a human hepatoma cell line (PLC/PRF/5) have been isolated by molecular cloning and have been partially sequenced. The HBV sequences are fragmented and rearranged. Thus, the surface antigen gene is the only intact HBV transcription unit present in these integrated sequences. The sites of integration recombination are dispersed over the entire viral genome; there is some preference for integration within the double-stranded portion of the genome. There are no repeats at the ends of the integrated HBV DNA fragments. Thus, recombination does not take place in a manner resembling the integration of retroviruses. The sequence data suggest that each HBV fragment is of the adw subtype. However, the integrated DNAs show an unexpected degree of sequence divergence. Direct evidence for the duplication, transposition, and subsequent divergence of two sequences is presented. The data surprisingly suggest that infection-integration of four distinct adw strains occurred. PMID- 2983099 TI - Selectivity of interferon action in simian virus 40-transformed cells superinfected with simian virus 40. AB - Treatment of African green monkey kidney CV-1 cells with human alpha interferons before infection with simian virus 40 (SV40) inhibited the accumulation of SV40 mRNAs and SV40 T-antigen (Tag). This inhibition persisted as long as the interferons were present in the medium. SV40-transformed human SV80 cells and mouse SV3T3-38 cells express Tag, and interferon treatment of these cells did not affect this expression. SV80 and SV3T3-38 cells which had been exposed to interferons were infected with a viable SV40 deletion mutant (SV40 dl1263) that codes for a truncated Tag. Exposure to interferons inhibited the accumulation of the truncated Tag (specified by the infecting virus) but had no significant effect on the accumulation of the endogenous Tag (specified by the SV40 DNA integrated into the cellular genome). The level of Tag in SV40-transformed mouse SV101 cells was not significantly decreased by interferon treatment. SV40 was rescued from SV101 cells and used to infect interferon-treated and control African green monkey kidney Vero cells. Tag accumulation was inhibited in the cells which had been treated with interferons before infection. Our data demonstrate that even within the same cell the interferon system can discriminate between expression of a gene in the SV40 viral genome and expression of the same gene integrated into a host chromosome. PMID- 2983100 TI - Role of antibody in primary and recurrent herpes simplex virus infection. AB - When herpes simplex virus was inoculated into the flank of a BALB/c mouse by scarification, the local replication of virus was followed by the establishment of an acute ganglionic infection. The subsequent centrifugal spread of this virus along nerves to the skin of the whole dermatome led to the development of a bandlike "zosteriform" rash. This represents a highly reproducible system in which virus travels through the nervous system synchronously in large numbers of animals. The transection of peripheral nerves at various times after infection showed that the virus had completed the round trip 60 h after inoculation into the upper flank and was detectable as infectious virus by 74 h postinfection. After the administration of virus, neutralizing but not nonneutralizing antibodies prevented the development of the zosteriform rash. The target epitopes of the protective antibodies were not confined to a single glycoprotein. Neutralizing antibody was effective even when given up to 60 h postinfection and was protective even when administered after sensory neurotomy at this time. Antibody was therefore able to prevent clinically and virologically detectable infection of the skin, presumably by acting peripherally on virus emerging from nerve endings. A quantitative estimate of the action of one of the neutralizing monoclonal antibody preparations, AP7, showed that high titers (several times higher than those normally found in immune mice) were needed to prevent this type of infection. These results are discussed in relation to antibody prophylaxis. PMID- 2983101 TI - Reassortant rotaviruses as potential live rotavirus vaccine candidates. AB - A series of reassortants was isolated from coinfection of cell cultures with a wild-type animal rotavirus and a "noncultivatable" human rotavirus. Wild-type bovine rotavirus (UK strain) was reassorted with human rotavirus strains D, DS-1, and P; wild-type rhesus rotavirus was reassorted with human rotavirus strains D and DS-1. The D, DS-1, and P strains represent human rotavirus serotypes 1, 2, and 3, respectively. Monospecific antiserum (to bovine rotavirus, NCDV strain) or a set of monoclonal antibodies to the major outer capsid neutralization glycoprotein, VP7 (of the rhesus rotavirus), was used to select for reassortants with human rotavirus neutralization specificity. This selection technique yielded many reassortants which received only the gene segment coding for the major neutralization protein from the human rotavirus parent, whereas the remaining genes were derived from the animal rotavirus parent. Single human rotavirus gene substitution reassortants of this sort represent potential live vaccine strains. PMID- 2983102 TI - Genetic analysis of bovine papillomavirus type 1 trans-acting replication factors. AB - The establishment of bovine papillomavirus type 1 in somatic mammalian cells is mediated by extrachromosomal replication and stable maintenance of the viral genome as a multicopy nuclear plasmid. Previous studies indicated the requirement of viral gene expression for bovine papillomavirus type 1 replication and plasmid maintenance (M. Lusky and M. R. Botchan, Cell 36:391-401, 1984; Turek et al., Proc. Natl. Acad. Sci. U.S.A. 79:7914-7918, 1982). To define the viral genes which are necessary for this process, we constructed a series of specific mutations within the viral genome and assayed the resulting mutants for their ability to replicate extrachromosomally in mouse C127 cells. We report here that the bovine papillomavirus type 1 trans-acting replication factors were encoded by at least two distinct viral genes since the mutants fell into two complementation groups, rep and cop. Mutants (rep-) affecting the E1 open reading frame (ORF) failed to replicate bovine papillomavirus type 1 DNA extrachromosomally and would integrate into chromosomal DNA. We suggest that this gene product is one of the factors required to specifically preclude the integration event. Mutants (cop-) affecting the E7 ORF were maintained in the extrachromosomal state; however, the copy number of the mutant genomes was reduced 100-fold compared with that of wild type DNA. Analysis of single-cell subclones showed that each cell contained the mutant genomes at a copy number of one to two, indicating that the cop- phenotype did not reflect a simple segregation defect. We propose that the gene defined by mutations in the E7 ORF played a crucial role in stably maintaining the copy number of the viral plasmid at high levels. Genomes with mutations in the cop and rep complementation groups, when cotransfected, rescued the wild-type phenotype, extrachromosomal replication with a high, stable copy number for both types of plasmids. Therefore, the gene products acted in trans, and the mutations were recessive to the wild-type functions. One specific rep- mutant showed a 30-fold increased transformation efficiency when compared with that of the wild-type genome. In addition, morphological transformation mediated by the cop- mutants appeared to be unstable. These results imply that either or both of the replication functions played some role in regulating the expression of the viral transforming functions. PMID- 2983104 TI - A fragment of the simian virus 40 early genome can induce tumors in nude mice. AB - Cell lines transformed by simian virus 40 mutant F8dl (deleted from 0.168 to 0.424 map units, corresponding to the carboxy-terminal 62% of the wild-type simian virus 40 large tumor antigen) are tumorigenic in nude mice. Four of five C3H10T1/2 cell lines transformed by F8dl were tumorigenic in nude mice, whereas two of two wild-type transformants were tumorigenic. PMID- 2983103 TI - Nucleotide sequence analysis of the BALB/c murine sarcoma virus transforming gene. AB - We determined the nucleotide sequence of the v-H-ras-related oncogene of BALB/c murine sarcoma virus. This oncogene contains an open reading frame of 189 amino acids that initiates and terminates entirely within the mouse cell-derived ras sequence. The protein encoded by this open reading frame matches the sequence predicted for the T24 human bladder carcinoma oncogene product, p21, in all but two positions. The presence of a lysine residue in position 12 of BALB/c murine sarcoma virus p21 likely accounts for its oncogenic properties. PMID- 2983105 TI - Map location of homologous regions between Marek's disease virus and herpesvirus of turkey and the absence of detectable homology in the putative tumor-inducing gene. AB - The DNA region having homology between Marek's disease virus and herpesvirus of turkey was assigned to the restriction map of Marek's disease virus by Southern blot hybridization. Under moderate conditions at the level of 15% mismatching, homology was found to be distributed throughout the Marek's disease virus genome. The long inverted-repeat regions (TRL and IRL), which are considered to play a significant role in tumorigenicity, did not show any homology to herpesvirus of turkey DNA. PMID- 2983106 TI - Characterization of an endogenous retrovirus-repetitive DNA chimera in the mouse genome. AB - We found that an endogenous mouse mammary tumor provirus, GR-MTV-8, is embedded within a member of the BAM HI family of long interspersed repetitive mouse DNAs. GR-MTV-8 appears to be transcriptionally silent at its normal chromosomal position in the mouse genome. The provirus is transcriptionally active, however, when cloned and transfected into mouse cells (Kennedy et al., Nature (London) 295:622-624, 1982). We propose that the transcriptional inactivity of GR-MTV-8 in situ is due to an inhibitory effect, possibly involving DNA methylation, attributable to the flanking BAM HI element. PMID- 2983107 TI - Genetic analysis of the susceptibility of mouse cytomegalovirus to acyclovir. AB - Eight independently derived mouse cytomegalovirus (MCMV) mutants resistant to acyclovir (ACV) were obtained by the sequential plating of wild-type virus in increasing concentrations of ACV. Results of complementation studies among these eight mutants suggest that all had mutations within the same or closely associated genes. A ninth MCMV mutant resistant to phosphonoacetate (PAA) derived by plating wild-type virus in the presence of 100 micrograms of PAA per ml displayed coresistance to ACV and was unable to complement any of the ACV-derived mutants. Recombination experiments among all combinations of the nine MCMV mutants were performed and supported the complementation data in that no recombination could be detected. Seven of the eight ACV-resistant mutants demonstrated cross-resistance to PAA and hypersensitivity to aphidicolin. The one mutant not coresistant to PAA was more susceptible to PAA than was the parent virus. Only a few mutants demonstrated coresistance when the mutants were tested against 9-beta-D-arabinofuranosyladenine (ara-A). The ACV mutant that demonstrated increased susceptibility to PAA was 30-fold more susceptible to ara A but remained unchanged in susceptibility to aphidicolin. Two of the parent mutant combinations were selected for DNA synthesis analysis in the presence of ACV (5 microM). A significant decrease in DNA synthesis was demonstrated for both parent viruses, and there was little effect on mutant virus DNA synthesis at the same drug concentration. These results suggest that susceptibility of MCMV to ACV is confined to a product of a single gene and that a mutation of this gene can lead to an altered phenotype when compared with parent virus in susceptibility of DNA synthesis to PAA, ara-A, and aphidicolin, drugs that are known to inhibit DNA polymerase activity. PMID- 2983108 TI - The minimum transforming region of v-abl is the segment encoding protein-tyrosine kinase. AB - Only 1.2 kilobases (kb) at the 5' end of the 3.9-kb v-abl sequence in Abelson murine leukemia virus is required for fibroblast transformation. A precise delineation of this minimum transforming region was made by using small 5' or 3' deletions. Insertions of four amino acids, generated by putting synthetic DNA linkers into various restriction enzyme cleavage sites, abolished transforming activity, indicating that much of the internal sequence of the minimum transforming region plays a critical role in the transformation process. This 5' 1.2 kb of v-abl encodes protein-tyrosine kinase activity when expressed in Escherichia coli. Each of the mutations which caused a loss of transformation activity also resulted in a loss of protein-tyrosine kinase activity when expressed in E. coli. The minimum transforming region of v-abl contains amino acid homology to other protein-tyrosine kinase oncogenes, and a comparison with these oncogenes is presented. PMID- 2983109 TI - Protein stabilization explains the gag requirement for transformation of lymphoid cells by Abelson murine leukemia virus. AB - The single protein encoded by Abelson murine leukemia virus is a fusion of sequence from the retroviral gag genes with the v-abl sequence. Deletion of most of the gag region from the transforming protein results in a virus capable of transforming fibroblasts but no longer capable of transforming lymphoid cells. Smaller deletions in gag reveal that p15 gag sequences are responsible for this effect, whereas deletion of p12 sequences had no effect on lymphoid transformation. In transformed fibroblasts, p15-deleted and normal proteins had similar activities and subcellular localization. When the p15-deleted genome was introduced into previously transformed lymphoid lines, its protein product exhibited a marked instability. The tyrosine-specific autophosphorylation activity per cell was less than 1/20th that of the nondeleted protein. Although pulse-Ia-beling showed that the p15-deleted protein was synthesized efficiently, immunoblotting demonstrated that its steady-state level was less than 1/10th that of the nondeleted Abelson protein. The specific instability of the p15-deleted protein in lymphoid cells explains the requirement of these sequences for lymphoid but not fibroblast transformation. PMID- 2983110 TI - Generation of glucocorticoid-responsive Moloney murine leukemia virus by insertion of regulatory sequences from murine mammary tumor virus into the long terminal repeat. AB - The glucocorticoid-regulatory sequences from the murine mammary tumor virus long terminal repeat (MMTV LTR) were introduced into the LTR of Moloney murine leukemia virus (M-MuLV) by recombinant DNA techniques. The site of insertion was in the M-MuLV LTR U3 region at -150 base pairs with respect to the RNA cap site. Infectious M-MuLVs carrying the altered LTRs (Mo + MMTV M-MuLVs) were recovered by transfection of proviral clones into NIH-3T3 cells. The Mo + MMTV M-MuLVs were hormonally responsive in that infection was 3 logs more efficient when performed in the presence of dexamethasone, irrespective of the orientation of the inserted MMTV sequences. However, even in the presence of hormone, the Mo + MMTV M-MuLVs were less infectious than wild-type M-MuLV. In contrast to the large effect on infectivity, dexamethasone induced virus-specific RNA levels in chronically Mo + MMTV M-MuLV-infected cells only two- to fourfold. Fusion plasmids between the altered LTRs and the bacterial chloramphenicol acetyltransferase gene allowed the investigation of LTR promoter strength by the transient chloramphenicol acetyltransferase expression assay. The chloramphenicol acetyltransferase assays indicated that the insertion of MMTV sequences into the M-MuLV LTR reduced promoter activity in the absence of glucocorticoids but that promoter activity could be induced two- to fivefold by dexamethasone. The Mo + MMTV M-MuLVs were also tested for the possibility that viral DNA synthesis or integration during initial infection was enhanced by dexamethasone. However, no significant difference was detected between cultures infected in the presence or absence of hormone. The insertion of MMTV sequences into an M-MuLV LTR deleted of its enhancer sequences did not yield infectious virus or active promoters, even in the presence of dexamethasone. PMID- 2983111 TI - In vitro transcription analysis of the viral promoter involved in c-myc activation in chicken B lymphomas: detection and mapping of two RNA initiation sites within the reticuloendotheliosis virus long terminal repeat. AB - Chicken syncytial virus, a member of the reticuloendotheliosis virus family, induces B-cell lymphomas in chickens that arise by transcriptional activation of the chicken c-myc gene. In vitro transcription studies on cloned tumor DNA containing a deleted chicken syncytial virus provirus integrated upstream from, and in the same transcriptional orientation as, the chicken c-myc coding region were utilized to map possible transcriptional promoters and initiation sites. In vitro transcripts extending into c-myc sequences were initiated at two sites within the downstream long terminal repeat (LTR) closest to c-myc coding sequences. Both initiation sites have been precisely mapped by S1 nuclease and DNA sequencing methods. One site (I1) lies at the U3-R junction of the LTR, and the other site (I2) lies approximately 160 nucleotides upstream. Transcriptional control signals, including TATA- and CAAT-like sequences are present at appropriate distances upstream from the initiation sites. Both initiation sites are utilized to a similar extent. The upstream chicken syncytial virus LTR was also shown to be transcriptionally active in vitro. Two strong transcriptional initiation sites were also found in the LTR of spleen necrosis virus, a related member of the reticuloendotheliosis virus family; therefore, it seems likely that the existence of two transcriptional initiation sites is a common feature of the reticuloendotheliosis virus LTR, in contrast to other previously studied retroviral LTRs that exhibit one such site. The possible implications of these findings are discussed. PMID- 2983112 TI - Molecular cloning of two paralytogenic, temperature-sensitive mutants, ts1 and ts7, and the parental wild-type Moloney murine leukemia virus. AB - ts1 and ts7, the paralytogenic, temperature-sensitive mutants of Moloney murine leukemia virus (MoMuLV), together with their wild-type parent, MoMuLV-TB, were molecularly cloned. ts1-19, ts7-22, and wt-25, the infectious viruses obtained on transfection to NIH/3T3 cells of the lambda Charon 21A recombinants of ts1, ts7, and wt, were found to have retained the characteristics of their non-molecularly cloned parents. In contrast to the wt virus, ts1-19 and ts7-22 are temperature sensitive, inefficient in the intracellular processing of Pr80env at the restrictive temperature, and able to induce paralysis in CFW/D mice. Like the non molecularly cloned ts7, the ts7-22 virion was also shown to be heat labile. The heat lability of the ts7 virion distinguishes it from ts1. Endonuclease restriction mapping with 11 endonucleases demonstrated that the base composition of MoMuLV-TB differs from that of the standard MoMuLV, but no difference was detected between the molecularly cloned ts1 and ts7 genomes. However, ts1 and ts7 differ from MoMuLV in the loss or acquisition of four different restriction sites, whereas they differ from MoMuLV-TB in the loss or acquisition of three different restriction sites. PMID- 2983113 TI - Detection and physical map of a omega tox+-related defective prophage in Corynebacterium diphtheriae Belfanti 1030(-)tox-. AB - A library of chromosomal DNA from Corynebacterium diphtheriae Belfanti 1030(-)tox was cloned in the lambda phage vector EMBL4 and screened for sequences homologous to corynephage omega tox+ and the attB1-attB2 region of the C7(-)tox- chromosome. Two portions of the 1030(-)tox- chromosome, 35 and 30.5 kilobases long which contain, respectively, the entire region homologous to corynephage omega tox+ and the attB1-attB2 sites, were mapped with the restriction endonucleases BamHI and EcoRI. Chromosomal DNA from 1030(-)tox- was shown to contain a 15.5-kilobase region that was homologous to ca. 42% of the corynephage omega tox+ genome. These sequences were found to hybridize to three regions of the phage genome and do not contain either the diphtheria tox operon or the attP site. These sequences are distant from the chromosomal region that contains the attB1-attB2 sites. Moreover, unlike other known defective prophages, the physical map of this prophage starts at the cos site and is colinear with the vegetative phage map. The 30.5-kilobase region of the 1030(-)tox- chromosome, which contains the attB1-attB2 sites, has a central core region that is almost identical to the corresponding region of the C7(-)tox- chromosome; however, the flanking sequences in these two strains of C. diphtheriae are different. PMID- 2983114 TI - Both trans-acting factors and chromatin structure are involved in the regulation of transcription from the early and late promoters in simian virus 40 chromosomes. AB - We isolated simian virus 40 (SV40) chromosomes from lytically infected CV-1 cells at various times during the late phase and transcribed them in vitro with either whole-cell or nuclear extracts of HeLa cells. The late promoter was 3- to 10-fold more active than the early promoter. With bare SV40 DNA templates, the early promoter was up to 10-fold stronger than the late promoter. The relative strengths of the early and late promoters on SV40 chromosomes were essentially independent of template concentration or length of the replicative phase of the infection. When monoclonal antibodies or antisera against T antigen (T Ag) were added to SV40 chromosomes or when T Ag, both free and chromatin bound, was removed by immunoprecipitation with anti-T, the activity of the late promoter remained essentially unchanged. Washing with 0.4 M NaCl removed T Ag from more than 90% of the mature chromosomes associated with T Ag. Transcription from the late promoter still predominated in the salt-washed T Ag-depleted chromosomes, even though there was a marked increase in early promoter activity. The depression of the early promoter could be reversed by adding the T Ag-containing extract back to the depleted chromosomes. Extraction of SV40 chromosomes with 1.5 M NaCl resulted in a decrease in the activity of the late promoter and a further increase in the activity of the early promoter so that the relative amounts of early and late RNA synthesized were similar to those for bare SV40 DNA templates. Late RNA synthesis from bare SV40 DNA templates was stimulated by high-speed supernatants prepared from nuclear extracts of SV40-infected cells but not from those of uninfected cells. Pretreatment of the supernatants with anti-T did not alter the result. Our findings indicate that the activity of the early and late SV40 promoters is regulated by at least two different mechanisms at the chromosomal level. One is mediated by a subclass of T Ag bound to SV40 chromosomes which represses early SV40 transcription but has no effect on late transcription. A second level of regulation, involving a tightly bound trans acting chromosomal factor and a stable nucleoprotein structure, favors the late promoter over the early promoter by up to 10-fold. PMID- 2983115 TI - Mapping and sequence of the gene for the pseudorabies virus glycoprotein which accumulates in the medium of infected cells. AB - RNA from pseudorabies virus (PRV)-infected cells was translated in a reticulocyte lysate with and without the addition of dog pancreas microsomes. Upon addition of the microsomes to the translation reaction, an additional prominent protein product was observed that was not present when microsomes were omitted. The gene coding for this processed protein and its lower-molecular-weight precursor was mapped within the small unique region of the genome by hybridization of mRNA to cloned fragments of PRV DNA and translation of the selected mRNAs. A fragment of the coding region of this gene was inserted into an open reading frame cloning vector to express part of this gene as a hybrid protein in Escherichia coli. This hybrid protein was injected into mice to raise an antiserum which was found to precipitate the glycoprotein which accumulates in the medium of PRV-infected cells. This allows us to conclude that the gene for the "excreted" glycoprotein (gX) maps to the small unique region of the genome, and that the precursor of this glycoprotein is readily processed by dog pancreas microsomes. The region of the PRV genome which codes for this glycoprotein was sequenced and found to include an open reading frame coding for 498 amino acids, flanked by sequences which contain features common to eucaryotic promoters and polyadenylation signals. The predicted protein sequence includes a hydrophobic sequence at the N terminus which could be a signal sequence, and a hydrophobic sequence followed by a hydrophilic sequence at the C-terminus. PMID- 2983116 TI - Different membrane anchors allow the Semliki Forest virus spike subunit E2 to reach the cell surface. AB - The Semliki Forest virus spike subunit E2, a membrane-spanning protein, was transported to the plasma membrane in BHK cells after its carboxy terminus, including the intramembranous and cytoplasmic portions, was replaced by respective fragments of either the vesicular stomatitis virus glycoprotein or the fowl plague virus hemagglutinin. The hybrid proteins were constructed by cDNA fusion. Upon a transient expression they could be localized at the cell surface by immunofluorescence with specific antibodies directed against any of the protein fragments. PMID- 2983117 TI - Cytomegalovirus early and late membrane antigens detected by antibodies in human convalescent sera. AB - Using indirect immunofluorescence and a panel of human convalescent-phase sera, we identified cytomegalovirus (CMV) early and late membrane antigens (CMV-EMA and CMV-LMA, respectively) as separate entities on the surfaces of viable CMV infected fibroblasts starting at 6 to 12 and 36 to 48 h postinoculation, respectively. For expression of CMV-EMA and CMV-LMA, infectious virus and active protein synthesis were required, whereas the expression of CMV-LMA, in addition, required viral DNA synthesis. Our data suggest that CMV-EMA and CMV-LMA form an individual set of CMV antigens that are different from intracellular CMV antigens and possibly (partly) different from the viral envelope. PMID- 2983118 TI - P3HR-1 Epstein-Barr virus with heterogeneous DNA is an independent replicon maintained by cell-to-cell spread. AB - We present results of biological experiments which indicate that the subpopulation of Epstein-Barr virus strain P3HR-1 with heterogeneous (het) DNA consists of self-contained replicons which multiply alongside, but independently of, Epstein-Barr virus strain HR-1 containing standard DNA. When a population of HR-1 virions containing het DNA was introduced into X50-7 cells, the input heterogeneous DNA increased in abundance, as did the DNA of the endogenous virus of X50-7 cells. The input standard HR-1 viral DNA, however, was not amplified. When parental HR-1 cells or a cellular subclone containing het DNA were grown for several weeks in the presence of human serum with neutralizing antibody, the het DNA was lost from the culture; standard HR-1 DNA, however, was not affected by antiserum. Furthermore, virions containing het DNA could be serially propagated through cellular subclones of HR-1 cells which lack het DNA. After each serial passage, cells which acquired het DNA released virions with the ability to induce early antigens in Raji cells. These experiments define a novel in vitro life cycle of an Epstein-Barr virus variant which is maintained, not vertically by partitioning to daughter cells in cell division, but horizontally by cell-to-cell spread. PMID- 2983119 TI - Cytotoxic T-lymphocyte reactivity with individual Sendai virus glycoproteins. AB - Liposomes were constituted with affinity-purified Sendai virus glycoproteins HN and F and phosphatidylcholine (PC) or phosphatidylethanolamine: phosphatidylserine (PEPS). The glycoprotein-bearing recombinant vesicles (RV) were used to modify the surface of P815 mastocytoma cells (H-2d) or EL4 lymphoma cells (H-2b). The cells treated with HN-F-PCRV, HN-PEPSRV, or F-PEPSRV were shown by surface immunofluorescence to retain antigen for at least 2 h at 37 degrees C after treatment. The modified cells were used in cytotoxicity assays with effector spleen cells from either DBA/2 (H-2d) or C57BL/6 (H-2b) immunized by inoculation of active Sendai virus. Cells modified by treatment with HN/F-PCRV showed susceptibility to cytolysis similar to that in actively infected cells. Cells modified with HN-PEPSRV or with F-PEPSRV were also susceptible. The sum of reactivities of the anti-HN component and the anti-F components was close to that seen with HN- and F-bearing targets. Syngeneic but not allogeneic target cells expressing Sendai virus glycoproteins were bound and lysed by the effector cells, which was expected if the interactions were major histocompatibility complex restricted. The activity was attributed to cytotoxic T lymphocytes, since it was depleted by treatment with anti-Thy 1.2 antibody and complement. PMID- 2983120 TI - Protection against rotavirus-induced gastroenteritis in a murine model by passively acquired gastrointestinal but not circulating antibodies. AB - Newborn mice suckled on dams immunized either orally or parenterally with primate rotavirus SA-11 were protected against diarrhea induced by SA-11 virus challenge. Experimental oral administration of milk from orally immunized dams protected suckling mice against challenge; protective activity was detected both in the anti-rotavirus immunoglobulin A (IgA) and IgG fractions, but IgA was more potent in vivo than IgG. Oral administration of milk from parentally immunized dams also protected suckling mice against challenge; in this case, protective activity was detected in the anti-rotavirus IgG fraction. In newborn mice foster-nursed by seronegative dams, circulating rotavirus-specific antibodies in high titer did not protect mice against oral SA-11 virus challenge. It appears that the most effective rotavirus vaccine will be that which induces an efficient production of antibodies active at the intestinal cell surface. PMID- 2983121 TI - A monoclonal antibody that neutralizes poliovirus by cross-linking virions. AB - The neutralization of type 1 poliovirus by monoclonal antibody 35-1f4 was studied. The virions were rapidly linked by antibody into oligomers and larger aggregates, followed by slow redistribution of antibody between the immune complexes. The antibody content and infectivity of immune complexes were determined. Remaining single virions were fully infectious and free of antibody. The oligomers and larger aggregates did not significantly contribute to the residual infectivity, which therefore correlated with the number of remaining single virions. Papain digestion of neutralized poliovirus released fully infectious, antibody-free virions from the immune complexes. Anti-immunoglobulin antibodies reneutralized these virions. Polymerization was shown to occur even at virus concentrations of less than 10(3) PFU per ml. PMID- 2983122 TI - Posttranscriptional control of DHFR gene expression during adenovirus 2 infection. AB - The effects of productive adenovirus infection on host gene expression were studied by using a line of methotrexate-resistant HeLa cells with amplified dihydrofolate reductase (DHFR) genes. We have previously reported that synthesis of DHFR is induced threefold early in infection and is shut off late in infection (Yoder et al., Mol. Cell. Biol. 3:819-828, 1983). These changes in DHFR protein synthesis are accompanied by changes in both the steady-state cytoplasmic levels of DHFR mRNA and in the rate of appearance of DHFR mRNA in the cytoplasm. In this report, we examined the mechanism of nuclear control of DHFR mRNA levels. Transcription of DHFR-specific sequences continued at a constant rate throughout infection, representing 0.015% of the total transcriptional activity. In contrast, nuclear steady-state levels of DHFR sequences changed in correspondence to the changing rate of appearance of DHFR mRNA in the cytoplasm. That is, nuclear levels of DHFR-specific sequences rose 2.5-fold early in infection and declined to a level below that found in uninfected cells late in infection. Thus, the relative nuclear stability of DHFR sequences changed throughout the course of infection such that during the time of induction, DHFR sequences were preferentially stabilized. This stabilization was transient, however, and was no longer observed by the time of shutoff. These data indicate that posttranscriptional nuclear events are important in the regulation of DHFR gene expression by adenovirus. PMID- 2983123 TI - Infectious hepatitis A virus particles produced in cell culture consist of three distinct types with different buoyant densities in CsCl. AB - Although hepatitis A virus (HAV) released by infected BS-C-1 cells banded predominantly at 1.325 g/cm3 (major component) in CsCl, smaller proportions of infectious virions banded at 1.42 g/cm3 (dense HAV particles) and at 1.27 g/cm3 (previously unrecognized light HAV particles). cDNA-RNA hybridization confirmed the banding of viral RNA at each density, and immune electron microscopy demonstrated apparently complete viral particles in each peak fraction. The ratio of the infectivity (radioimmunofocus assay) titer to the antigen (radioimmunoassay) titer of the major component was approximately 15-fold greater than that of dense HAV particles and 4-fold that of light HAV particles. After extraction with chloroform, the buoyant density of light and major component HAV particles remained unchanged, indicating that the lower density of the light particles was not due to association with lipids. Light particles also banded at a lower density (1.21 g/cm3) in metrizamide than did the major component (1.31 g/cm3). Dense HAV particles, detected by subsequent centrifugation in CsCl, were indistinguishable from the major component when first banded in metrizamide (1.31 g/cm3). However, dense HAV particles recovered from CsCl subsequently banded at 1.37 g/cm3 in metrizamide. Electrophoresis of virion RNA under denaturing conditions demonstrated that dense, major-component, and light HAV particles all contained RNA of similar length. Thus, infectious HAV particles released by BS-C 1 cells in vitro consist of three distinct types which band at substantially different densities in CsC1, suggesting different capsid structures with varied permeability to cesium or different degrees of hydration. PMID- 2983124 TI - Human leukocytes kill varicella-zoster virus-infected fibroblasts in the presence of murine monoclonal antibodies to virus-specific glycoproteins. AB - Seven murine monoclonal antibodies reacting with major glycoproteins of varicella zoster virus were tested for functional activity in assays for antibody-dependent cellular cytotoxicity (ADCC) and antibody-plus-complement-mediated lysis. Human peripheral blood mononuclear cells killed varicella-zoster virus-infected fibroblasts in the presence of three of four monoclonal antibodies directed against gp98/62 and a single monoclonal antibody directed against gp118. Neither of two monoclonal antibodies directed against gp66 was able to mediate ADCC. In 18-h assays, adherent effector cells were more active than nonadherent effector cells in mediating ADCC. Adherent cells treated with anti-Leu-11b and complement retained their cytotoxic activity, suggesting that monocytes are responsible for most of the adherent-cell-mediated cytotoxicity. Both immunoglobulin G1 and G2a murine monoclonal antibodies were able to participate in ADCC. Of the two immunoglobulin G2a monoclonal antibodies tested, both of which reacted with gp98/62, only one mediated lysis in the presence of complement. These results indicate that some murine monoclonal antibodies against major glycoproteins of varicella-zoster virus have functional activity in cytotoxicity assays. PMID- 2983125 TI - Intratubular malignant germ cells (carcinoma in situ) accompanying invasive testicular germ cell tumors. AB - We examined 111 radical orchiectomy specimens from patients with germ cell tumors of the testis treated between 1971 and 1981 for the presence of intratubular malignant germ cells (so-called carcinoma in situ) in adjacent testicular tissue. Of the specimens 12 were unevaluable because the entire specimen was tumor, while 36 of 44 cases of pure seminoma (82 per cent) and 41 of 55 mixed or nonseminomatous tumors (75 per cent) had intratubular malignant germ cells. Of the 22 cases without intratubular malignant germ cells 10 had severe atypia that suggested an earlier or borderline stage of development. These data support the contention that germ cell tumors originate as intratubular neoplasms or so-called carcinoma in situ in the seminiferous tubules. PMID- 2983126 TI - Ultrastructure of vesicourethral innervation. III. Axoaxonal synapses between postganglionic cholinergic axons and probably SIF-cell derived processes in the feline lissosphincter. AB - Ultrastructurally nonspecialized axoaxonal-type synapses between postganglionic cholinergic axons innervating the lissosphincter and apposed probably SIF-cell derived neuronal processes are described. These processes were considered as the peripheral extraganglionic counterparts of those belonging to type I SIF cell interneurons, described in many mammalian peripheral autonomic ganglia. On this basis, the herein reported cholinergic/probably SIF-cell derived axoaxonal-type synapses are proposed as an auxiliary mechanism of prejunctional catecholaminergic inhibition of the normally excitatory cholinergic (postganglionic parasympathetic) neuroeffector transmission in the feline lissosphincter. PMID- 2983127 TI - Frequent transmission of HTLV-III among spouses of patients with AIDS-related complex and AIDS. AB - Seven cases of acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) associated with human T-cell leukemia (lymphotropic) virus (HTLV)-III infection were documented in married males whose family members consented to medical evaluation. Five spouses also had evidence of HTLV-III infection documented by isolation of virus and by detection of serum antibody directed against viral structural proteins. Three of the five HTLV-positive spouses also had clinical evidence of ARC. The remaining four spouses were clinically healthy. Eleven children (between 14 months and 13 years of age) were also evaluated for evidence of disease and exposure to HTLV-III. Only one 14-month-old child was seropositive for antibody to HTLV-III. These data support the opinion that close household contact to patients with ARC or AIDS is not an efficient mechanism for virus transmission, while demonstrating that HTLV-III can be transmitted by repeated heterosexual contact. PMID- 2983128 TI - Use of AIDS antibody test may provide more answers. PMID- 2983129 TI - Zoster after exposure to varicella-zoster virus. PMID- 2983131 TI - Leads from the MMWR. Preventing lead poisoning in young children--United States. PMID- 2983130 TI - Isolation of lymphadenopathy-associated virus (LAV) and detection of LAV antibodies from US patients with AIDS. AB - A human retrovirus was isolated from the peripheral blood of three American patients newly diagnosed with the acquired immunodeficiency syndrome (AIDS). In each case the major core viral protein (p25) was shown to be antigenically identical to that of the prototype lymphadenopathy-associated virus (LAV). Two of the viral isolates were derived from intravenous narcotics abusers, the first demonstration of LAV isolation from this risk group. Antibody to LAV was detected by an IgG enzyme-linked immunosorbent assay in the serum samples of these and 14 additional American patients with AIDS and in none of 12 hospital worker controls. These findings provide support for the etiologic association of LAV and AIDS. PMID- 2983132 TI - Development and early natural history of HTLV-III antibodies in persons with hemophilia. AB - Antibodies to human T-cell lymphotropic virus type III (HTLV-III) were first detected in 1979 in serum samples from 30 known seropositive patients with hemophilia, over half of whom seroconverted in 1981-1982. Lymphadenopathy was present in 70% who were seropositive more than three years, compared with 10% who were seropositive three years or less. T-helper cell counts were low (307 +/- 64 cells/cu mm) in the early seroconverters, and normal in the late seroconverters. T-suppressor cell counts were not related to the year of seroconversion. The long latency period after seroconversion suggests an ongoing indolent process, rather than an acute infection. It remains to be determined whether this is an aberrant part of the immune response initiated by HTLV-III antigens or the result of a chronic active HTLV-III infection. PMID- 2983133 TI - Studies on collagen in the experimental myocardial infarction. AB - The healing process of myocardial infarction was studied with special reference to the production of collagen by the determinations of prolylhydroxylase (PHase) activity and contents of hydroxyproline (Hyp) of the collagen subfractions, i.e., neutral salt soluble (NSC), acid soluble (ASC) and insoluble collagens (ISC) in comparison with histologic observation of the proliferation of connective tissue in the canine heart. The contents of Hyp in NSC and ASC increased in the infarcted tissue 5 days after coronary ligation prior to its increase in ISC. The content of Hyp in NSC, which is a precursor of ASC and ISC, reached the maximal value at 7 days and that reached the maximal value in ASC 7 to 14 days after coronary ligation. ISC increased markedly after 14 days. PHase activity increased on the second day and attained to the highest at 5 to 7 days. Proliferation of fibroblasts paralleled with the increases in PHase activity and NSC, and an increase in collagen fiber did with the contents of ISC. From these results it is concluded that in the healing stage of myocardial infarction, PHase was activated earlier than the increase in immature collagen in the infarcted area which was observed 5 to 7 days after coronary ligation and was followed by the production of mature collagen after 14 days. PMID- 2983134 TI - Hepatitis B virus and the prevention of primary cancer of the liver. PMID- 2983135 TI - Cytogenetic studies in human T-cell lymphoma virus (HTLV)-positive leukemia lymphoma in the United States. AB - Cytogenetic studies were conducted on fresh and cultured cells from 11 patients with human T-cell leukemia virus-associated adult T-cell leukemia-lymphoma. Clones with abnormal karyotypes were detected in 9 of the 11 patients. Chromosome numbers were near-diploid in cells from all but 1 patient who also had a tetraploid clone. The chromosome abnormalities in these cells were extensive; numerous complex structural changes were seen in every chromosome pair. Structural abnormalities occurred most frequently in chromosome 6. The 6 patients with chromosome 6 deletions had breakpoints at bands q11, q13, q16q23, q21q23, q22q24, and q23q24. The characteristic clinical features of these 6 patients were aggressive course, short survival, poor response to chemotherapy, high white blood cell counts, hypercalcemia, and bone lesions, whereas cytogenetically abnormal patients without chromosome 6q deletions tended to have a more indolent course. The precise role of the 6q deletion cannot be established with certainty from these data. However, this abnormality appears to occur with a greater than expected frequency in this large cell aggressive lymphoma, in association with hypercalcemia and lytic bone lesions. PMID- 2983136 TI - Comparative studies of purified subviral component vaccines and formalin-treated mammary tumor viruses from four inbred strains of mice. AB - Formalin-treated virus vaccines were prepared from purified murine mammary tumor viruses (MuMTV) from 4 inbred strains of mice: RIII/Imr, GR/Imr, C3H/Imr, and A/Imr. In addition, subviral components were isolated from these 4 strains and purified to homogeneity. The inactivated viruses, their major envelope glycoproteins (gp50-gp55), and their major internal core protein (p28) were emulsified in complete Freund's adjuvant and used as vaccines for prevention of mammary tumors in mice. All 4 Formalin-treated virus vaccines reduced significantly the incidence of mammary tumors in "virus-free" C57BL and BALB/c mice when inoculated prior to challenge with live MuMTV. The RIII-, GR-, and A MuMTV strains showed extensive heterologous cross-protection, whereas the C3H MuMTV strain showed significant protection only against C3H- and A-MuMTV challenge. The major viral glycoproteins gp50-gp55 reduced significantly the tumor incidence when mice were challenged with isologous infectious virus after immunization, although these glycoproteins showed different degrees of cross protection than did the same virus strains used as "intact" but Formalin-treated preparations. RIII-gp55 and GR-gp55 cross-protected against each other but not against challenge with C3H- and A-MuMTV strains; the A-gp50 protected against challenge with A- and RIII-MuMTV strains; C3H-gp55 demonstrated limited activity against C3H-MuMTV challenge only. The internal viral core proteins (p28) were ineffective in all systems studied. The same vaccines were tested in MuMTV positive, high-tumor-incidence strains from which they were derived. At best, the appearance of spontaneous tumors was delayed in a few experimental sets; eventually, all mice developed mammary tumors. The foster-nursed C3HfC57BL strain of mice, which is not exposed to exogenous MuMTV during suckling and which develops mammary tumors after activation of the endogenous virus genome later in life, was responsive only when the heterologous GR-MuMTV Formalin-treated vaccine was used. The association between the ability of virus vaccines to protect a mouse strain and the degree of natural virus expression in that strain is discussed. PMID- 2983137 TI - Preventive and enhancing effects of retinoids on the development of naturally occurring tumors of skin, prostate gland, and endocrine pancreas in aged male ACI/segHapBR rats. AB - The effects of dietary retinoids on the development of naturally occurring tumors in retired breeder male ACI/segHapBR rats were investigated. Groups of rats (21 25 mo of age, an age when early neoplasms first appear and tumor incidences are generally low) were fed diets containing 1 of 3 retinoids--all-trans-N-4-(4 hydroxyphenyl)retinamide (4-HPR), 783 mg/kg diet; all-trans-N-(4 pivaloyloxyphenyl)retinamide (4-PPR), 951 mg/kg; or all-trans-4-N-(2 hydroxyethyl)retinamide (2-HER), 687 mg/kg--or control diet for up to 54 weeks (average, 33 wk). Rats were maintained until less than 20% remained and the experiment was terminated. Contributing causes of death were determined, and a complete necropsy was performed for each rat. There was no difference between the retinoid-treated rats and control rats in the average age at death (30-31 mo) or in the average experimental survival time (29-35 wk), in the proportions of tumor bearing rats (95.6-100%), or in the average number of organs with tumor per rat (2.1-2.5). The incidences of pancreatic islet cell adenoma and skin tumors were significantly different between control and some retinoid-treated groups. 4-PPR and 2-HER significantly enhanced pancreatic islet cell adenoma yields (P less than .025 and 0.05, respectively) whereas 4-HPR significantly inhibited epithelial and connective tissue skin tumor yields (P less than .025). Incidences of skin and prostate tumors were lower than in controls, but not significantly, in rats receiving 4-PPR and 2-HER. Most of the islet cell adenomas were shown, by avidin-biotin-peroxidase complex immunocytochemistry, to be insulinomas. 4-HPR would seem to be the most effective retinoid in the group, inasmuch as it prevented skin tumor development, may have slightly decreased the incidence of prostate tumors, and did not enhance islet cell tumor incidence. PMID- 2983138 TI - Globoside and Forssman synthases in human lymphocytes exposed to Epstein-Barr virus and mitogens. AB - The activities of two glycolipid synthetases, globoside synthase or UDP-N acetylgalactosamine-trihexosylceramide beta-N-acetylgalactosaminyltransferase (beta-GalNAc transferase; EC 2.4.1.79) and Forssman synthase or UDP-N acetylgalactosamine-globoside-alpha-N-acetylgalactosaminyltransfer ase (alpha GalNAc transferase; EC 2.4.1.88), were assayed in various human lymphoblastic cell lines. The activity of beta-GalNAc transferase was much higher than that of alpha-GalNAc transferase except in Molt 3 and Molt 4 lines, which were derived from T-cells. In cultivated human peripheral lymphocytes concanavalin A (Con A), lipopolysaccharide (LPS), and Epstein-Barr virus (EBV) stimulated the activities of alpha- and beta-GalNAc transferases in addition to having their known stimulative effect on thymidine incorporation. Characteristic differences between alpha- and beta-GalNAc transferases were noted in the responses to the above mitogens, but activities of both enzymes were greatly increased by exposure of the lymphocytes to EBV. Treatment of lymphocytes with either dactinomycin (actinomycin D) or cycloheximide 24 hours after the addition of Con A, LPS, or EBV decreased the activities of the transferases. This observation suggests that stimulation of alpha- and beta-GalNAc transferases requires transcriptional and translational processes. PMID- 2983139 TI - Ultrastructure and distribution of intracellular spicules in rat lung following chronic tobacco smoke exposure. AB - Factors that contribute to the induction of lung tumors in F344 rats by chronic tobacco smoke exposure were studied. Previous studies had shown that tobacco smoke exposures of 1-2 years duration induced only one major type of lesion involving the respiratory airways, i.e., fibrotic and cellular enlargement of peribronchiolar alveolar septa. The airway epithelium in these areas was metaplastic and in some of the lesions, the airway lining epithelium advanced out onto the surfaces of adjacent alveoli. Epithelial cells in these lesions frequently contained elongated cytoplasmic inclusions which were oriented with their long dimensions roughly in the same plane as the long axis of the cell. Macrophages contained similar but larger inclusions. Because the composition of the inclusions could be indicative of their origin, we subjected samples of treated and control lung tissues concurrently to transmission electron microscopy and energy-dispersive X-ray fluorescence spectrometry. Spectra from inclusions of macrophages indicated the presence of the elements sulfur, phosphorus, aluminum, silicon, and iron. Spectra from type II cells, however, which did not contain inclusions, showed a different elemental composition. The results suggested that spicules were present in epithelial cells throughout the airways. Minor lesions corresponding to "microinvasion" of epithelium into the lamina propria and of capillaries into the epithelial layer were also found in the trachea. PMID- 2983141 TI - Antibodies to bovine leukemia virus and presence of malignant lymphoma in slaughtered California dairy cattle. AB - Rates of malignant lymphoma (ML) in California dairy cattle and relationships between ML and presence of antibodies to bovine leukemia virus (BLV) were investigated. An excessively high rate of lymphoma was found in the population studied and a previously unknown relationship between ML and p-24 antibodies to BLV was identified. Sera from slaughtered California holstein dairy cows were tested by agar gel immunodiffusion for presence of antibodies to the gp-51 and p 24 antigens of BLV. Of the 7,760 cows slaughtered, 32% and 10% had gp-51 and p-24 antibodies, respectively, and 52 cows (0.67%) were condemned for ML. Lymphoma was diagnosed histopathologically in 37 (86%) of 43 cows sampled from those 52 condemned. Highly elevated prevalence rates of ML in gp-51 and p-24 antibody positive cows were 172.6 and 511.5 per 10,000 cows, respectively. A prevalence rate of lymphoma not associated with presence of BLV antibodies was estimated for the first time to be 1.55 per 10,000 cows. A highly significant (P less than .00001) and strong (odds ratio = 26.0) association was found between presence of p-24 antibodies and presence of ML for cows with gp-51 antibodies. When results of serologic tests were compared in regard to diagnosis of ML, the highest sensitivity was with use of gp-51 serology alone (97.3%) and highest specificity was with use of p-24 serology alone (90.4%). False-negative and false-positive rates of diagnosis of ML for gp-51-positive, p-24-negative and gp-51-positive, p 24-positive cows were 0.21 and 94.8%, respectively. PMID- 2983140 TI - Murine in vitro antigenic modification of tumor cells: effect on susceptibility to natural cell-mediated cytotoxicity. AB - A 3-methylcholanthrene [(MCA) CAS: 56-49-5]-induced fibrosarcoma cell line and its Friend murine leukemia virus-infected counterpart were assessed for their susceptibility to lysis by so-called "natural" effector cells in a series of 51Cr release assays. Detailed functional and phenotypic analysis of lytic effector cell populations from normal C57BL/6 mouse spleens revealed an identity most closely associated with natural cytotoxic cells. Neither tumor cell line was found to be sensitive to natural killer-mediated lysis. Additionally, virus infection of the MCA-induced fibrosarcoma cell line did not affect susceptibility to natural cell-mediated cytotoxicity. PMID- 2983142 TI - [Polyvinylpyrrolidone iodine and arginase: effect on corneal regeneration and its antiviral effect]. AB - PVP-iodine was found to be virucidal for adenoviruses and herpes simplex virus in a concentration of 0.5%, i.e., a 1:20 dilution of Betaisodona. Similar concentrations did not delay the healing of corneal epithelium of the rabbit after experimental lesions (Ullerich and Durchschlag, 1953), although the eyes were slightly irritated. However, a higher concentration (1.25%) caused considerable ocular irritation, and healing of the cornea was delayed or completely inhibited. Arginase, which inhibited the multiplication of adenoviruses and herpes simplex virus in cell culture, did not impair corneal healing in solutions of up to 5%. PMID- 2983143 TI - [How effective is the treatment schedule in children with infantile spasms?]. AB - A fatal incident during combined anticonvulsive therapy with ACTH and Dexamethason prompted us to reevaluate our treatment results in 19 children suffering from infantile spasms. We followed Doose's recommendation using Clonazepam as a means of first choice. Failing that, than Dexamethason and Primidone was used. A combination of ACTH, Dexamethason and Primidone was administered to patients failing both treatment courses. Our results question the necessity, the tolerance limit and the efficacy of this treatment schedule. Therefore we propose a prospective multicentric study to reinvestigate the therapeutic guideliness for infantile spasms. PMID- 2983145 TI - [Holoprosencephaly--clinical picture and genetic counseling in 6 cases]. AB - Inadequate midfacial and forebrain development may result from constitutional chromosome aberrations, gene defects and, possibly, from as yet unidentified terato-genetic agents. The spectrum of clinical manifestations ranges from cyclopia with grossly incomplete organogenesis of the forebrain to apparently minor deviation from normal midface morphogenesis presenting as hypotelorism or absence of the philtrum. Such minor facial dysmorphias may, however, be likewise accompanied by severe anomaly in brain development and function. We report six cases of holoprosencephaly defects in children without demonstrable chromosomal anomalies. The presenting clinical symptoms in these cases were anomalies of cranio-facial shape, hypotelorism; nasal and ocular malformations, as well as median clefts. Some cases presented additional defects in extra craniofacial regions. Two infants who survived for several hours showed evidence of forebrain defects on CT-scans. Three of the cases suggest autosomal-recessive inheritance with 25% recurrence risk on the basis of proven or highly probable parental consanguinity. The remaining, presumably sporadic cases carry a low empirical recurrence risk. Three of the six cases received direct or indirect hormone treatments during early pregnancy. PMID- 2983144 TI - [Sonographic detection of brain tumors in infancy]. AB - Using the open fontanelle as an acoustic window brain tumours were diagnosed by gray scale ultrasonography in 3 infants aged 1 day to 5 months. The tumours were characterized by their echo dense structure and their good delimination from the surrounding brain. In 2 children the tumour was localized infratentorially (medulloblastoma and unclassified neuroectodermal tumour) and had caused an occlusive hydrocephalus. Both children died aged 3 and 5 months in central nervous dysregulation. One child suffered from plexus papilloma which had caused a hypersecretory hydrocephalus. After resection of the tumour the hydrocephalus decreased without any further treatment. Comparison with axial computed tomography and autopsy findings showed, that gray scale ultrasonography is equally efficient in diagnosing brain tumours and associated hydrocephalus. PMID- 2983147 TI - [Skin injury after exposure to high-energy protons and helium ions]. AB - The RBE of high-energy protons and helium ions was measured with respect to the proliferation rate, cell number of the epidermic basal layer, induction rate of aberrant mitoses, number and type of chromosome aberrations. The animals were examined on postradiation days 1 and 6 (i.e., when the proliferation rate of the first postradiation mitosis was at a maximum). The RBE coefficients of protons and helium ions were unity with respect to the reproductive cell death and cell number of the epidermic basal layer; 1.5 with respect to the formation of multiple cell aberrations; 1.2-1.5 with respect to exchange-type aberrations; and 1.6 with respect to the mitotic index variations. This gives evidence that in response to high-energy protons the nuclear apparatus of the cell undergoes profound lesions. PMID- 2983146 TI - [Adrenoleukodystrophy in a pair of siblings]. AB - The diagnosis of adrenoleukodystrophy (ALD) in two brothers was confirmed by the analysis of long-chain fatty acids in cultured skin fibroblasts. The 23 year old brother was treated for Addison's disease at the age of 7 years. His first symptoms of ALD developed at the age of 22. These were lack of concentration and compulsive disorder. The younger brother was noted to show behavioural changes and a decreased performance at school at age 9. The disease then progressed rapidly. Variability and diagnostic procedures are discussed in this report. PMID- 2983149 TI - Aspiration pneumonia with adenovirus infection in a Japanese macaque (Macaca fuscata fuscata). AB - Pneumonia with aspiration of foreign bodies and adenovirus infection was identified in a wild Japanese macaque which died 1 month after capture. The adenovirus infection was considered to be secondary in importance to the pneumonia. PMID- 2983148 TI - [Effect of space flight factors on the hormonal regulation of water-salt metabolism]. AB - This paper presents the results of examinations of 19 test subjects exposed to head-down tilting at -8 and -15 degrees and of 14 test subjects kept in water immersion for 24 hours. During the first hours of exposure the renal excretion of water and monovalent ions increased. Renin and aldosterone measurements showed that changes in the sodium and potassium excretion were produced by a lower activity of the renin-angiotensin-aldosterone system in the first 1.5 hour of hypokinesia. During immersion the renal excretion of calcium and magnesium also grew, especially in the evening and at night. The PTH production and calcium concentration in blood increased, thus augmenting the nephron load. The diurnal rhythms of the renal excretion of potassium, calcium and magnesium remained unchanged and those of water, osmotically active substances and sodium varied. The data obtained indicate significant changes in water-salt metabolism and its regulation within the first hours of head-down tilt and water immersion. PMID- 2983150 TI - A comparison of human breast cancer cell kinetics measured by flow cytometry and thymidine labeling. AB - Flow cytometric determination of tumor ploidy and S-phase fraction following collagenase dissociation and thymidine labeling was performed on 75 consecutive breast cancers. Estrogen and progesterone receptor levels and routine histologic examination also were obtained on each tumor. Cell viability following collagenase dissociation varied from 13 to 95% with a mean of 71%. Thirty-six tumors were diploid, four tetraploid, and four hypertetraploid, and the remainder had DNA indices between 1.1 and 1.9. There was no significant correlation between tumor ploidy and tumor size or estrogen receptor positivity or negativity. The percentage of cells in S-phase varied from 1.2 to 20.0% with a mean of 6.0% utilizing a rectilinear model for histogram analysis that integrated a 10 contiguous channel sample containing the lowest number of cells in S-phase (S pFL). The mean S-pFL of diploid carcinomas (3.43%) was significantly lower than that of hyperdiploid carcinomas (8.38%). There was good correlation between S phase fraction determined by thymidine-labeling index (TLI) and S-pFL (r = 0.772, p = 0.0001). S-pFL predicted whether a tumor would be above or below median TLI with an accuracy of 90.5%. Estrogen receptor-negative cancers tended to have higher TLIs and S-pFLs than estrogen receptor-positive cancers; however, there was no correlation between progesterone receptor positivity or negativity and TLI and S-pFL. PMID- 2983151 TI - Levamisole in the treatment of glioblastoma multiforme. AB - Twenty-five patients with malignant glioma were randomized to receive radiation therapy only or radiation therapy and oral Levamisole. There were no differences in survival between the two groups and therefore no advantage in the use of Levamisole in glioblastoma could be demonstrated. PMID- 2983152 TI - Nasopharyngeal carcinoma due to Epstein-Barr virus: serologic tests. PMID- 2983153 TI - [Vasculitis caused by hypersensitivity in HBsAG-negative liver cirrhosis]. PMID- 2983154 TI - [Malignant hyperthermia]. PMID- 2983155 TI - [A new generation of broad-spectrum antibacterial chemotherapeutic agents belonging to the group of DNA gyrase inhibitors (derived from pyridine carboxylic acid or quinolines)]. PMID- 2983156 TI - Immunologic aspects of otologic disease: an overview. AB - The immunologic explosion has now reached the field of otology. By having better techniques to measure the changes at cellular and molecular levels, it is now possible to devise experiments to show morphologic anatomic changes as well as functional changes. The demonstration in 1980 (M.S.) that tympanosclerosis could be induced immunologically represents a concrete advancement in immunologic thinking in conceptualization of otologic disease. In 1974, one of the authors (M.S.) published work dealing with the treatment of vasculitis of immunologic origin for sudden hearing loss. This was aimed at inhibiting the complement cascade from starting its destructive action. Recently, the immunologic challenge in animals demonstrated by changes in the inner ear was shown by one of the authors (T.J.Y.). Such changes were compatible with labyrinthine hydrops, or Meniere's disease, otosclerosis, and sensorineural hearing loss and vestibular dysfunction. PMID- 2983157 TI - Glomus jugulare tumors revisited: a ten-year statistical follow-up of 231 cases. AB - Glomus jugulare tumors are rather common in the middle ear and temporal bone. They are usually easy to diagnose and surgically remove. However, there are a comparatively large number of these growths which not only cause extensive local destruction, but can spread distally and even have endocrine manifestations. This group is difficult to diagnose and manage despite recent advances in diagnostic procedures and the advent of skull base surgery. There is a void in the literature of a large series of cases being followed for a long period of time. This paper presents a comprehensive detailed statistical ten-year follow-up of 231 glomus jugulare tumors. The results suggest that our present diagnostic procedures often are inadequate in finding small tumors, and our treatment and follow-up are lacking because these tumors tend to recur often at a late date and in a different form. A comprehensive method of approach of diagnosis and treatment is described to manage all cases of glomus jugulare. PMID- 2983158 TI - Effect of chronic alcohol consumption on rat brain microsome lipid composition, membrane fluidity and Na+-K+-ATPase activity. AB - The present study reports differences in phospholipid classes, fatty acids of individual phospholipids, and changes in membrane fluidity and Na+-K+-ATPase activity in brain microsomes of rats maintained on an alcohol diet for 35 days compared to sex, age and weight-matched control rats maintained on a calorically equivalent, non-alcohol diet. Although no difference in Na+-K+-ATPase activity was found in microsomes from alcohol vs control rats when measured in the absence of added alcohol, the presence of low concentrations of ethanol (less than 100 mM) stimulated, while high concentrations (greater than 100 mM) inhibited enzyme activity. The stimulation was differentially expressed in that the microsomal enzyme from alcohol rats was stimulated to a lesser extent than the enzyme from control rats. However, the inhibiting effect of high concentrations of alcohol was similar in microsomes from both alcohol and control rats. Also in membranes from alcohol rats, there was a lower quantity of phosphatidylethanolamine (PE) and higher quantities of phosphatidylserine (PS) and phosphatidylinositol (PI) compared to membranes from control rats. The major change in fatty acid composition was a reduction in the level of polyunsaturated fatty acids, which was particularly evident in PI and PS. The linoleic acid: arachidonic acid ratio (18:2/20:4) and the saturation:unsaturation ratio were also increased in PI and PS in membranes from alcohol animals. However, the ratio of n-6/n-3 fatty acids remained the same or was reduced in membranes from alcoholic animals. Although no difference in the inherent "fluidity" of membranes from alcohol vs control rats could be demonstrated by electron paramagnetic resonance, molecular tolerance to ethanol was demonstrated in the membranes from alcohol rats by the resistance to the disordering effects of added ethanol. PMID- 2983159 TI - Dietary chloride as a determinant of disordered calcium metabolism in salt dependent hypertension. AB - In rats given desoxycorticosterone (DOC), the recently reported finding that a normal amount of dietary sodium chloride (NaCl) induces hypertension but an equimolar amount of sodium bicarbonate (NaHCO3) does not, might be a consequence of the differing effects of the two sodium salts on the metabolism of calcium. In accord with this hypothesis, we have found that, in uninephrectomized rats given DOC: Dietary NaCl induces persisting hypercalciuria and hypertension whereas an approximately equimolar amount of dietary NaHCO3 induces neither hypercalciuria nor hypertension. The urinary excretion of calcium becomes greater in rats given NaCl than in those given NaHCO3, before their blood pressures become different. Replacing dietary NaCl with a near equimolar amount of dietary NaHCO3 corrects both the hypercalciuria and the hypertension initially induced by NaCl. PMID- 2983160 TI - Cholecystokinin receptor mediated hydrolysis of inositol phospholipids in guinea pig gastric glands. AB - CCK-octapeptide (CCK-8) (EC50 = 0.5 nM), in the presence of Li+, increased 3H inositol phosphate (IP) accumulation in guinea pig gastric glands prelabeled with 3H-inositol. CCK-8 desulfate, human gastrin I and pentagastrin were much less potent than CCK-8. Antagonists of CCK receptors such as proglumide, dibutyryl-c GMP and CBZ-Tyr (SO3H)-Met-Gly-Trp-Met-AspNH2 shifted the CCK dose response curve to the right. However, histamine (H1 and H2), cholinergic, substance P and alpha- and beta-adrenergic receptor antagonists had no effect on 3H-IP accumulation induced by CCK. The results suggest that CCK receptor activation in gastric glands leads to an enhanced breakdown of inositol phospholipids which may relate to calcium mobilization and pepsinogen secretion. PMID- 2983161 TI - Interaction of PHM, PHI and 24-glutamine PHI with human VIP receptors from colonic epithelium: comparison with rat intestinal receptors. AB - PHM, the human counterpart of porcine Peptide Histidine Isoleucine amide (PHI), is shown to be a VIP agonist with low potency on human VIP receptors located in colonic epithelial cell membranes. Its potency is identical to that of PHI but by 3 orders of magnitude lower than that of VIP itself in inhibiting 125I-VIP binding and in stimulating adenylate cyclase activity. This contrasts markedly with the behaviour of PHI on rat VIP receptors located in intestinal epithelial cell membranes where PHI is a potent agonist with a potency that is 1/5 that of VIP. In another connection, we show that 24-glutamine PHI has the same affinity as 24-glutamic acid PHI (the natural peptide) for rat or human VIP receptors. These results indicate that while PHI may exert some physiological function through its interaction with VIP receptors in rodents, its human counterpart PHM is a very poor agonist of VIP in human. Furthermore, they show that the drastic change in position 24 of PHI (neutral versus acid residue) does not affect the activity of PHI, at least on VIP receptors. PMID- 2983162 TI - One- and two-receptor models of opioid drug action. AB - The history of models for opioid drug receptors is traced and two currently proposed models are compared, the first envisaging two rigid receptors, one for agonists and one for antagonists, while the second model postulates one single flexible receptor which in its native conformation fits opioid antagonists but is distorted when it binds to agonists. Important recent experiments are satisfactorily interpreted by both models but the flexible one-receptor model requires fewer assumptions and explains more facts. PMID- 2983163 TI - Urea-induced myoclonus: medullary glycine antagonism as mechanism of action. AB - Stimulus sensitive myoclonus is a prominent symptom of uremia in both man and animals. Intravenous injection of urea into cats had been previously reported to produce spike and sharp wave electrical discharges in the medullary reticular formation which correlated with the myoclonic movements. In the present investigations, intraperitoneal injections of 2 g/kg urea every 15 minutes for 4 injections produced myoclonus in rats accompanied by brain urea concentrations of 6.8 X 10(-2)M, which is sevenfold higher than normal. 10(-2) and 10(-1) M urea significantly reduced 3H-strychnine binding to rat medulla membranes by 30% and 43% respectively. Urea inhibition of 3H-strychnine binding was reversible and binding kinetics revealed that 10(-1)M urea decreased Bmax by 65% with no effect on the affinity. Brain glycine levels did not change after urea injections and urea had no effect on synaptosomal uptake of 3H-glycine. Urea did not alter 3H GABA, 3H-glutamate and 3H-QNB receptor binding but decreased 3H-diazepam receptor binding in the medulla. Mannitol also reduced 3H-diazepam binding but had no effect on 3H-strychnine binding. Stereotaxic injection of the glycine receptor antagonist, strychnine, into the rat medullary reticular formation produced myoclonus, whereas Ro 15-1788, a benzodiazepine antagonist, had no effect. Urea may produce myoclonus by blockade of glycine receptors in the medullary reticular formation. PMID- 2983164 TI - Opposite effects of two ligands for peripheral type benzodiazepine binding sites, PK 11195 and RO5-4864, in a conflict situation in the rat. AB - The effects of two drugs acting at the peripheral type benzodiazepine binding sites, PK 11195 and RO5-4864, were examined in shock-induced suppression of drinking in rats. These two compounds have opposite effects : RO5-4864 (3.1-1205 mg/kg i.p.) enhanced whereas PK 11195 (25-50 mg/kg i.p.) decreased the punished responding, and PK 11195 (6.25 mg/kg, a dose which did not alter the punished responding) blocked the proconflict action of RO5-4864 (6.25 and 12.5 mg/kg). The effects of RO5-4864 and PK 11195 were not antagonized by RO15-1788, a selective antagonist of the central benzodiazepine site. In addition, PK 11195 (6.25 mg/kg) did not reverse the proconflict effect of two beta-carbolines : beta-CEE and FG 7142. AS picrotoxin did not change the punished responding, these data imply that the effects of RO5-4864 and PK 11195 on the one hand and those of chlordiazepoxide and beta-carbolines on the other hand are differentially mediated and suggest that the peripheral type benzodiazepine binding sites are involved in this conflict model. PMID- 2983165 TI - Restraint stress enhances morphine-induced analgesia in the rat without changing apparent affinity of receptor. AB - Antagonism of morphine analgesia (tail-flick assay) by naloxone was assessed quantitatively by in vivo "apparent" pA2 determination in unstressed rats and in rats subjected to restraint stress. Restrained rats had a higher baseline tail flick latency than did unstressed (unrestrained) animals, and were more sensitive to the analgesic effect of morphine, as reflected in lower morphine ED50s. There was no significant difference between apparent pA2 values of unstressed and restrained rats using pA2 regression line analysis. This suggests that while stress enhances the analgesic effect of morphine, it does not appreciably alter opiate receptor affinity for naloxone under the conditions of this study. PMID- 2983166 TI - Isolation and purification of morphine receptor by affinity chromatography. AB - Brain membranes were solubilized by sonication and Triton X-100 extraction and applied to an affinity column consisting of a 6-succinyl morphine derivative of Affi Gel-102. A fraction exhibiting high opiate binding was eluted by tris-buffer containing naloxone, CHAPS and NaCl. This fraction consisted of both proteins and acidic lipids. The opiate binding properties of this purified material exhibited many properties similar to those of membrane bound receptors of the u-type, including high affinity, stereospecificity, Na-effect and rank order in affinity for opiates. This opiate binding material was highly sensitive to both trypsin and N-ethylmaleimide. Based on the protein content of the isolated membrane receptor, a 3200-fold purification over the original brain P2 fraction was achieved. PMID- 2983167 TI - Hyperthermia induced by morphine administration to the VTA of the rat brain: an effect dissociable from morphine-induced reward and hyperactivity. AB - Morphine action at opiate receptors in the ventral tegmental area (VTA) of the rat brain has been implicated in the production of increased locomotor activity and in morphine's rewarding properties. In the present experiments, bilateral administration of morphine (18 micrograms tapped into the tips of 28 gauge cannulae) into the VTA resulted in an increase in body temperature in rats. This effect was both reversed and blocked by a systemic injection of the opiate receptor blocker, naloxone, suggesting that it was due to morphine action at opiate receptors. The neuroleptic, pimozide, injected systemically four hours prior to morphine administration completely blocked the increased locomotor activity but had no effect on the hyperthermia. These data demonstrate that the hyperthermia was not brought about by the increased physical activity. Furthermore, these results suggest that while morphine-induced reward and increased locomotor activity may be mediated by an interaction of morphine and the ascending mesolimbic dopamine system, the hyperthermia is not. In an additional experiment, the effect of systemic injections of the central neurotransmitter receptor antagonists, scopolamine, phenoxybenzamine, and methergoline, on the hyperthermia induced by morphine in the VTA was investigated. Only the serotonin antagonist, methergoline, attenuated the hyperthermia. PMID- 2983168 TI - Different effects of serotonin antagonists on 3H-mianserin and 3H-ketanserin recognition sites. AB - In minces prepared from the frontal cortex of rats treated with ketanserin (10 mg/kg i.p.) or mianserin (5 mg/kg i.p.) twice daily for 21 days, the Vmax of the adenylate cyclase stimulated by NE (100 microM) is attenuated, suggesting that ketanserin and mianserin share with a number of antidepressants the ability to attenuate the adenylate cyclase stimulation by NE. Ketanserin, given with the above mentioned dose schedule for 7 consecutive days, reduced the Bmax of 5HT2 recognition sites but failed to change either the Bmax or the apparent Kd of H mianserin binding. A significant decrease in the Bmax of 5HT2 binding sites is elicited also by a single injection of mianserin (1). This drug also down regulates its own binding when given twice daily for 3 weeks. From this and other information (2,3), it is concluded that ketanserin and mianserin bind to distinct recognition sites. The possibility that 5HT2 and mianserin recognition sites are functionally related and that serotonergic synapses are modulated by multiple chemical signals might be considered. PMID- 2983169 TI - Effect of GABA agonists on the neurotoxicity and anticonvulsant activity of benzodiazepines. AB - Progabide (50 mg/kg, i.p.), a GABA receptor agonist, significantly decreases the median minimal neurotoxic dose (TD50) of clobazam, chlordiazepoxide, and diazepam; the receptor binding of these substances is highly enhanced by muscimol. Progabide has no significant effect on the TD50 of clonazepam and triazolam; the receptor bindings of these substances is either only slightly enhanced or not altered by muscimol. Progabide also significantly decreases the median antimaximal electroshock dose (MES ED50) of all the benzodiazepines tested. However, progabide has no effect on the median antipentylenetetrazol dose (PTZ ED50) of the benzodiazepines. Likewise, THIP (2.5 mg/kg, i.p.) significantly decreases the TD50 of chlordiazepoxide but not that of triazolam. THIP significantly decreases the MES ED50 of chlordiazepoxide and triazolam but has no effect on the PTZ ED50 of these two substances. The above data suggest that benzodiazepine receptors linked to GABA receptors contribute to the minimal neurotoxicity and anti-MES activity but not to the anti-PTZ activity of benzodiazepines. PMID- 2983170 TI - Pressor and cardioaccelerator effects of gamma MSH and related peptides. AB - We have recently demonstrated that the hypertensinogenic and natriuretic actions of ACTHI-39 can be found in a non-steroidogenic fragment of ACTH, ACTH4-10. These effects of ACTH or ACTH4-10 may be due to their ability to act as weak agonists of gamma MSH. gamma MSH is found in the 16K N-terminus of pro-opiocortin, and contains a sequence analogous to ACTH4-10, gamma MSH3-9. We investigated the cardiovascular effects of gamma 2MSH, gamma MSH3-9, and sterically restricted analogs of ACTH4-10. The results indicate that gamma MSH3-9, had essentially the same activities as ACTH4-10. The addition of five other amino acid residues to gamma MSH3-9 (gamma 2MSH) resulted in significant enhancement of pressor and cardioaccelerator activity. Steric restriction of the ACTH4-10 sequence by the substitution of a D-Phe in place of an L-Phe residue in position #7, or cyclization of the peptide by a half-Cys4, half Cys10 intramolecular disulfide bridge derivatization, resulted in increased cardiovascular activities. Based on these data, the cardiovascular actions of ACTH4-10, gamma MSH3-9, and gamma 2MSH are predicted to be due to the assumption of a reverse-turn three-dimensional structure. The additional residues in gamma 2MSH appear to specifically enhance the cardiovascular activities of gamma MSH3-9. The results suggest the existence of a new class of hypophyseal peptides with cardiovascular activities, which require the assumption of a defined three-dimensional structure. PMID- 2983171 TI - TENA, a selective kappa opioid receptor antagonist. AB - A number of opioid antagonists (TENA, naloxone, Mr 2266, WIN 44441) were evaluated for their selectivity in antagonizing the effect of mu, kappa, and delta agonists in the guinea pig ileum (GPI) and mouse vas deferens (MVD) preparations. Among these four antagonists, TENA was the most potent and the only ligand which was selective for kappa receptors. In this regard TENA was approximately 27-times more effective in antagonizing the kappa agonist, U 50488H, relative to the mu agonist, morphine, and it was about 5-times more effective against ethylketazocine (EK) relative to morphine. At the same concentration (20 nM) TENA did not significantly antagonize the delta agonist, [D Ala2,D-Ala5]enkephalin (DADLE), in the MVD. Also, TENA was more effective than naloxone, EK, or U-50488H in protecting kappa receptors from irreversible blockage by beta-CNA. The results of this study indicate that TENA is the most selective kappa antagonist yet reported. PMID- 2983172 TI - Interaction of gastric inhibitory polypeptide (GIP) with the insulin-secreting pancreatic beta cell line, In lll: characteristics of GIP binding sites. AB - The interaction of GIP with its receptors in the hamster pancreatic insulin secreting beta cell line, In lll, has been analyzed. 125I-labelled GIP used as tracer showed the same affinity as native GIP for the GIP binding sites. Binding of the tracer was time, temperature and cell concentration dependent. It was saturable, reversible and highly specific. Under equilibrium conditions, i.e. 2 hours at 13 degrees C, 20% and 25% of the tracer and of GIP binding sites were inactivated, respectively. Native GIP inhibited binding of 125I-labelled GIP in a dose-dependent manner, saturation of the GIP binding sites being obtained at 3. 10(-7) M peptide. Two types of GIP binding sites were found by Scatchard analysis, a small population with a high affinity for GIP (KD = 7 nM) and a large population with a low affinity (KD = 800 nM). The biphasic dissociation process confirmed the GIP binding sites heterogeneity. Apart from GIP, no peptide tested influenced the binding of the 125I-labelled GIP. The present data represents the first analysis of functionally relevant GIP binding sites in a insulin-secreting cell. PMID- 2983173 TI - Involvement of opioid receptor subtypes in rat feeding behavior. AB - The short-acting opiate antagonist naloxone decreases food intake in three models of ingestive behavior: free feeding, food-deprivation induced feeding and deoxyglucose-induced feeding. Twenty-four hours after administration, the long acting, mu1 selective antagonist naloxonazine inhibits food intake to the same extent as naloxone in freely feeding and food-deprived rats, but not in animals treated with 2-deoxyglucose. These results indicate that 1) opiates modulate feeding through multiple opioid receptor mechanisms, one of which is the mu subtype, and 2) the feeding observed in various experimental paradigms are modulated by different receptor subtypes. Furthermore, these results illustrate the usefulness of naloxone in defining a behavior as opioid but point out its limitations in discriminating between opioid receptor subtypes. PMID- 2983174 TI - Effects of luteinizing hormone-releasing hormone agonist and calcium upon adenylyl cyclase activity of human corpus luteum membranes. AB - We have investigated the ability of the agonist analog of luteinizing hormone releasing hormone (LH-RH), D-Trp6-LH-RH (LH-RHa), and of CaCl2 to inhibit directly gonadotropin stimulation of adenylyl cyclase in a cell-free system prepared from human corpus luteum. In the presence of a submaximally effective concentration of hCG, addition of 10(-5)M final concentration of LH-RHa did not alter the gonadotropin-stimulated enzyme activity, nor did LH-RHa alone show any effect upon basal levels of the enzyme. The failure to inhibit adenylyl cyclase would indicate that the LH-RHa does not affect gonadotropin receptor binding or cAMP synthesis and/or degradation in this membrane system, suggesting that the luteolytic effects of LH-RH are unlikely to involve a direct antigonadotropic activity at the level of the human corpus luteum. In great contrast to LH-RHa, addition of CaCl2 resulted in a dose-dependent inhibition of hCG-stimulable adenylyl cyclase. Thus, in the presence of either a maximally or submaximally effective concentration of hCG, inhibition was significant at 0.5 mM CaCl2 added in excess of ATP (2 mM) and EDTA (1 mM), being about 90% upon addition of 2.5 mM CaCl2. We also found that calcium reduced enzyme stimulation by forskolin and the GTP analog, guanyl 5'-yl imidodiphosphate [GMP-P(NH)P] in a dose-related manner and that activation by NaF was less sensitive to inhibition by calcium. Accordingly, at 2.5 mM CaCl2, guanyl nucleotide and forskolin stimulations were inhibited 96% and 86%, respectively, while NaF stimulation was reduced by 40%. Because previous studies have shown that calcium does not impair gonadotropin binding activity, the calcium-dependent inhibition of gonadotropin responsiveness reported here would imply an alteration in the functional coupling of the components of the luteal adenylyl cyclase system. These data suggest that calcium may play a role in the regulation of gonadotropin action in the human corpus luteum. PMID- 2983175 TI - Cytomegalovirus infection in the normal host. AB - CMV mononucleosis often resembles EBV infectious mononucleosis; however, certain features of the history and physical may help to distinguish CMV from EBV. While CMV mononucleosis is usually self-limited, certain laboratory abnormalities may persist for months or years after the patient has recovered. Previous reports on CMV in the non-immunocompromised host have rarely described systemic complications. We have reviewed 10 cases of CMV with systemic manifestations at one institution over a 15-year period. These patients had prolonged fevers (often greater than three weeks) and the diagnosis was often unsuspected during the early part of the illness. While two patients required mechanical ventilation, all patients had self-limiting disease and survived. When CMV is suspected and diagnosed early in the course, numerous diagnostic (and potentially dangerous) tests can be avoided in a viral illness in which prolonged fever is common. PMID- 2983176 TI - Highly purified human growth hormone fails to stimulate lipolysis in rabbit adipocytes in vitro or in rabbits in vivo. AB - To determine whether the rapid lipolytic effect observed with human growth hormone (hGH) preparations in rabbits and rabbit adipose tissue is an intrinsic property of the hormone, we examined the lipolytic effects in vivo and in vitro of clinical grade preparations, hGH purified by DEAE cellulose chromatography, and hGH prepared by recombinant DNA techniques. Using isolated rabbit perirenal adipocytes, ACTH and clinical-grade hGH preparations both stimulated glycerol release to the same maximal rate with half-maximal hGH effects observed between 8 and 50 micrograms/mL. Purification of the most potent lipolytic preparation of clinical grade hGH by DEAE cellulose chromatography yielded a preparation (2 IU/mg of growth activity that retained insulinlike effects on rat fat pad [U-14C] glucose metabolism) that, at concentrations up to 0.2 mg/mL, failed to stimulate lipolysis by adipocytes incubated for one or four hours in the presence or absence of dexamethasone or trypsin inhibitor or when preincubated for three hours prior to addition of hGH. Recombinant DNA-derived hGH did not stimulate glycerol release at 0.1 mg/mL. While antiserum to purified hGH blocked the increase in glucose oxidation in rat fat pads produced by clinical grade hGH, it did not inhibit its lipolytic effect using rabbit adipocytes. Purified hGH (0.1 mg/kg IV) was also unable to elicit a rise in serum free fatty acid (FFA) levels of conscious rabbits while, at the same dose, clinical grade hGH increased FFA levels to 900 microEq/L over basal. Rapid lipolytic stimulation in rabbit adipocytes by hGH preparations could not be attributed to the 20,000 molecular weight variant of hGH (hGH20K) or the peptide corresponding to positions 32 to 46 of hGH (deletion peptide).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983177 TI - Sequestration and microsomal C-25 hydroxylation of [3H]-vitamin D3 by the rat liver. AB - A study of the vitamin D3 (D3) 25-hydroxylase was undertaken in an in vivo-in vitro model. [3H]-D3 (0.7, 1.0, 10, or 100 nmol/100 g of body weight) was injected into the portal vein and the liver was excised 18 seconds later. The liver homogenate was then submitted to differential centrifugation and the amount of [3H]-D3 incorporated in the subcellular fractions was evaluated. The microsomal fraction was also incubated in vitro and the appearance of [3H]-25 hydroxyvitamin D3 [25(OH)D3] was determined by high performance liquid chromatography (HPLC). Results showed that the fractional liver [3H]-D3 uptake varied between 37 percent and 48 percent of the dose injected. The intracellular distribution of [3H]-D3 showed that most of the vitamin was incorporated into the microsomal fraction (45% to 50% of the intracellular [3H]-D3) except at the highest dose of [3H]-D3 where the cytosolic fraction contained the highest amount (56.4%) of the incorporated vitamin. Mathematical analysis of the intracellular [3H]-D3 distribution showed that the microsomal fraction was the only subcellular fraction that was found to incorporate [3H]-D3 in relation to the total liver uptake of the vitamin. The apparent Michaelis-Menten kinetics of the [3H]-D3-25 hydroxylase showed that with substrate concentration of up to 88.5 nM, the apparent Km and Vmax were 28.2 nM and 25.8 fentomoles (fmol) X min-1 X mg microsomal pro-1, respectively, but the reaction lost considerable efficiency with higher substrate concentrations. With the in vivo-in vitro model used, the cytosolic fraction was not essential for the optimal C-25 hydroxylation of D3. These results show that the endoplasmic reticulum of rat hepatocytes possess a high capacity for D3 incorporation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983178 TI - Susceptibility of various cell culture systems to bovine viral diarrhea virus. AB - A comparative study was carried out on the susceptibility of primary bovine embryo kidney (pBEK) cell cultures and those of three other cell culture systems, i.e., primary calf testicle (pCT) cells and two cell lines, one of which originated from bovine embryonic trachea (EBTr) and the other from buffalo lung (IMR-31), to bovine viral diarrhea (BVD) virus. The virus titers obtained in the three culture systems under study did not differ significantly from those of pBEK cells. Moreover, the growth curve study demonstrated thad the pCT cells, as well as the two cell lines, were similar to the pBEK cultures in their susceptibility to the virus. Accordingly, it appeared reasonable to conclude that pCT, EBTr and IMR-31 cell cultures possess the general requirements for use in studies on the BVD virus. However, inasmuch as the cytopathic effects (CPE) produced by the EBTr and the IMR-31 cell lines did not display any distinctive characteristics typical of BVD virus-induced CPE, it is recommended that the two cell lines not be used for isolation of the virus from specimen material. PMID- 2983179 TI - A two-year longitudinal study on the etiology of acute diarrhea in young children in Northern Italy. AB - During a 24-month period, 561 young children (6 months-2 years of age) hospitalized for acute diarrhea were studied for enteric pathogens. Patients positive for one or more pathogens were 359 (64.0%). Infection with one pathogen was found in 266 (47.4%) patients, whereas multiple infection was detected in 93 (16.6%) patients. Enteropathogens associated with disease were Rotaviruses: 150 (26.7%), Adenoviruses: 99 (17,6%), non-polio Enteroviruses: 48 (8.5%), Coronaviruses: 10 (1.8%), Parvoviruses: 5 (0.9%), Salmonella sp.: 41 (7.3%), Campylobacter fetus: 45 (8.0%), Giardia lamblia: 8 (1.4%), and "enteropathogenic" E. coli: 63 (11.2%) of which 15 (3%) produced heat labile enterotoxin. Seasonal occurrence of enteropathogens is also described. PMID- 2983180 TI - Restricted replication of BK virus in human lymphocytes. AB - Human lymphocytes from either peripheral blood or continuous cultures (P3HR-1 cells) are able to support the replication of prototype polyomavirus BK (BKV) as well as its related strain BO15 virus (BO15V). Instead, human monocytes from peripheral blood, although able to bind and egulf BKV virions, do not express virus-specific antigens within a 50 day observation period. In the light of these results, a probable role is suggested for human mononuclear blood cells in the mechanism of natural infection by polyomaviruses. PMID- 2983181 TI - Sexuality and aging. PMID- 2983182 TI - Agonist-induced desensitization of muscarinic receptor-mediated calcium efflux without concomitant desensitization of phosphoinositide hydrolysis. AB - Phosphoinositide hydrolysis does not appear to desensitize in 1321N1 astrocytoma cells. The evidence for this is that 1) the rate of accumulation of [3H]inositol 1-phosphate is linear for up to 90 min in the presence of carbachol, 2) pretreatment of cells with 100 microM carbachol for 75 min does not diminish the subsequent ability of carbachol to increase [3H]inositol 1-phosphate accumulation, and 3) the production of all of the [3H]inositol phosphates including the polyphosphoinositide metabolites [3H]inositol bis- and trisphosphate continues for up to 75 min in the presence of carbachol and declines rapidly when the muscarinic receptor antagonist atropine is added. Only when cells are treated with carbachol for 2.5 hr or longer is there a reduction in carbachol-stimulated phosphoinositide hydrolysis, and this is associated with a decrease in muscarinic receptor number. There does appear to be desensitization of hormone-stimulated Ca2+ mobilization in 1321N1 cells, because treatment of these cells with carbachol for 75 min leads to loss of the subsequent ability of carbachol to stimulate unidirectional 45Ca2+ efflux. Histamine-stimulated 45Ca2+ efflux also is lost in cells pretreated with carbachol, indicating that the desensitization is heterologous. We conclude that desensitization of hormone stimulated, unidirectional 45Ca2+ efflux cannot be accounted for by a loss of receptor-mediated phosphoinositide hydrolysis. If phosphoinositide hydrolysis or inositol triphosphate formation are signals for calcium mobilization, the site at which the calcium response desensitizes must be distal to the initial receptor mediated activation of phospholipase C. PMID- 2983183 TI - Effects of nitroprusside, glyceryl trinitrate, and 8-bromo cyclic GMP on phosphorylase a formation and myosin light chain phosphorylation in rat aorta. AB - The effects of nitroprusside (NP), glyceryl trinitrate (GTN), and the 8-bromo analog of cyclic GMP (8-Br-cGMP) on norepinephrine (NE)-stimulated phosphorylase a formation and myosin light chain (MLC) phosphorylation were examined in the rat aorta. NE produced a time-dependent increase in tension, phosphorylase a formation, and MLC phosphorylation. The formation of phosphorylase a and phosphorylation of MLC were transient, since both processes declined to basal levels within 30 min after the addition of NE even though tension remained elevated. NP and GTN inhibited tension, phosphorylase a formation, and MLC phosphorylation although inhibition of phosphorylase was greater when strips were treated with submaximal (i.e., 0.01 microM) NE concentrations. GTN was a more effective inhibitor of phosphorylase a formation than NP in NE-treated strips, although both agents and 8-Br-cGMP inhibited MLC phosphorylation. The guanylate cyclase inhibitor methylene blue (10 microM) effectively prevented the effects of NP and GTN. The results suggest that NP, GTN, and 8-Br-cGMP inhibit phosphorylase kinase and MLC kinase activation by lowering Ca2+ in the cell. This hypothesis is supported by the observations that 8-Br-cGMP inhibited the Ca2+-dependent, KCl induced phosphorylase a formation most markedly at reduced concentrations of extra-cellular Ca2+. In addition, neither NP, GTN, nor 8-Br-cGMP inhibited phosphorylase a formation in forskolin-treated tissues, which occurred in response to cAMP-dependent phosphorylation of phosphorylase b kinase. PMID- 2983184 TI - Thiol-dependent DNA damage produced by anthracycline-iron complexes. The structure-activity relationships and molecular mechanisms. AB - Doxorubicin (Adriamycin) and daunomycin analogs have been examined for their ability to chelate iron and catalyze the oxidative cleavage of DNA. The results show that the C-11-hydroxyl group is essential for iron binding and DNA damage. Thus, the iron complexes of doxorubicin, daunomycin, carminomycin, and 4 demethoxydaunomycin are potent redox catalysts capable of reducing molecular oxygen in the presence of physiologic concentrations of glutathione. They are also effective catalysts of hydroxyl radical formation from hydrogen peroxide. With the exception of daunomycin, generation of hydroxyl radical from hydrogen peroxide is stimulated by greater than 200% by DNA addition. Analogs that lack the C-11-hydroxyl group are relatively inefficient at oxygen reduction, hydroxyl radical formation, and DNA cleavage. The potencies of the anthracycline analogs tested in the H2O2-dependent DNA cleavage reaction correlated well with their relative cardiac toxicities. PMID- 2983185 TI - Metabolic activation and toxicity of acetaminophen and related analogs. A theoretical study. AB - Reaction thermodynamics have been calculated for an oxene model for cytochrome P 450 oxidations of four related arylamines: aniline, p-hydroxyaniline, acetanilide, and acetaminophen, by both radical and nonradical mechanisms, using a semiempirical molecular orbital method (modified neglect of differential overlap). The results indicate that for both p-hydroxyaniline and acetaminophen, a recently proposed peroxidase-like mechanism leading directly to p benzoquinoneimines via radical intermediates is thermodynamically favored over N hydroxylamine formation by H abstraction or addition rearrangement. These studies also provide a detailed characterization of three candidate species for the toxic reactive intermediate of acetaminophen: 1) p-benzoquinoneimines, 2) the radical intermediate formed by H abstraction from the nitrogen, and 3) the radical intermediate formed by H abstraction from the phenol. Calculated electron and spin densities indicate that the radical formed by H abstraction from the phenol oxygen does not remain localized on the oxygen, but is primarily a semiquinone aryl radical with significant unpaired spin density on the ring carbon atoms, particularly on C-3 and C-5. This result is consistent with the hyperfine splitting pattern observed for a transient radical species in a hydroxyl radical mediated chemical oxidation of acetaminophen. The radical formed by H abstraction from the nitrogen also delocalizes on the ring carbons, but to a lesser extent and at the 2- and 4-positions. A closed shell mechanism of N oxidation of arylamines appears to lead directly to the hydroxylamines with less likelihood of precursor reactive intermediates. Toxic species could then be formed by loss of H2O from the hydroxylamines. PMID- 2983186 TI - Inheritance of extrachromosomal ribosomal DNA during the asexual life cycle of Dictyostelium discoideum: examination by use of DNA polymorphisms. AB - Wild-type isolates of Dictyostelium discoideum exhibited differences in the size of restriction fragments of the extrachromosomal 88-kilobase ribosomal DNA (rDNA) palindrome. Polymorphisms in rDNA also were found among strains derived solely from the NC4 wild-type isolate. These variations involved EcoRI fragments II, III, and V; they included loss of the EcoRI site separating fragments II and V and deletion and insertion of DNA. More than one rDNA form can coexist in the same diploid or haploid cell. However, one or another parental rDNA tended to predominate in diploids constructed, using the parasexual cycle, between haploid NC4-derived strains and haploid wild-type isolates. In some cases, most if not all of the rDNA of such diploids were of one form after ca. 50 generations of growth. Segregant haploids, derived from diploids that possessed predominantly a single rDNA allele, possessed the same allele as the diploid and did not recover the other form. This evidence implies that replication does not proceed from a single chromosomal or extrachromosomal copy of the rDNA during the asexual life cycle of D. discoideum. PMID- 2983188 TI - Characterization of a retrovirus shuttle vector capable of either proviral integration or extrachromosomal replication in mouse cells. AB - A retrovirus shuttle vector is described that contains the dominant selectable neo gene which confers resistance to kanamycin in bacteria and to the drug G418 in animal cells. The bacterial supF gene and the origins of DNA replication from polyomavirus and the ColE1 replicon also have been included in this vector. Infection of normal rodent cells results in single-copy proviral integration, whereas infection of mouse (MOP) cells expressing polyoma large T antigen results in extrachromosomal replication of the DNA form of the virus. The copy number of the extrachromosomal circles in MOP cells varies from 0 to 100 copies per cell. G418-resistant MOP cells lose their drug-resistant phenotype after passage under nonselective conditions, suggesting that maintenance of the extrachromosomal circles is unstable. The extrachromosomal form of the virus can be recovered as plasmids in Escherichia coli. Two-thirds of the circles analyzed were found to be structurally intact. The others have undergone rearrangements including deletions and insertions. The bacterial supF gene was found to be intact in the majority of recovered plasmids. The data presented here suggest that these retroviruses should be useful as gene transfer vectors for animal cells in culture or in vivo. PMID- 2983187 TI - Bacterial beta-galactosidase as a marker of Rous sarcoma virus gene expression and replication. AB - We have developed a convenient and sensitive assay of eucaryotic gene expression which uses the Escherichia coli lacZ gene product, beta-galactosidase, as a nonselectable marker. This system has been applied to the analysis of Rous sarcoma virus replication and gene expression. Avian cells were transfected with plasmids encoding in-frame gene fusions of the N-terminal portion of the gag gene to a 'lacZ gene, which requires both transcriptional and translational initiation signals; these were supplied by the virus long terminal repeat and leader region. Readily detectable quantities of beta-galactosidase were synthesized in transfected cells; it was demonstrated that the levels of enzyme activity induced in such cultures increased linearly with the input DNA concentration and also correlated with mRNA levels. By using a Rous sarcoma virus-derived vector containing the src gene and a related virus as a helper, it was shown that lac sequences were compatible with all phases of the virus life cycle. gag-lacZ fusion proteins were immunoprecipitable from cultures which stably expressed lacZ as well as src. Virus rescued from stably transfected cultures resulted in continued lac and src expression in recipient cells. One particular construction was efficiently transmitted as virus, although it lacked sequences thought to be important for encapsidation of RNA into virions. The data presented here demonstrate the use of lacZ as a marker of retrovirus gene expression and replication. PMID- 2983189 TI - Coordinate amplification of metallothionein I and II genes in cadmium-resistant Chinese hamster cells: implications for mechanisms regulating metallothionein gene expression. AB - We describe here the derivation, characterization, and use of clonal cadmium resistant (Cdr) strains of the Chinese hamster cell line CHO which differ in their metallothionein (MT) induction capacity. By nondenaturing polyacrylamide gel electrophoresis, we showed that the stable Cdr phenotype is correlated with the augmented expression of both isometallothioneins (MTI and MTII). In cells resistant to concentrations of CdCl2 exceeding 20 microM, coordinate amplification of genes encoding both isometallothioneins was demonstrated by using cDNA MT-coding sequence probes and probes specific for 3'-noncoding regions of Chinese hamster MTI and MTII genes. Molecular and in situ hybridization analyses supported close linkage of Chinese hamster MTI and MTII genes, which we have mapped previously to Chinese hamster chromosome 3. This suggests the existence of a functionally related MT gene cluster in this species. Amplified Cdr variants expressing abundant MT and their corresponding Cds parental CHO cells should be useful for future studies directed toward elucidating the mechanisms that regulate expression of the isometallothioneins. PMID- 2983190 TI - Transcription of herpes simplex virus tk sequences under the control of wild-type and mutant human RNA polymerase I promoters. AB - We studied RNA polymerase I transcription in cells transfected with a plasmid, prHuTK, containing the herpes simplex virus tk gene fused to a human rRNA promoter. Primer extension analysis of tk RNA isolated from COS cells transfected with prHuTK reveals that transcription from the RNA polymerase I promoter is highly efficient and initiates at the same position used for the synthesis of endogenous rRNA in HeLa cells. The RNA products derived from prHuTK are distinguishable from normal RNA polymerase II transcripts of tk in that they are not polyadenylated, are extremely unstable, and are found predominantly in the nucleus. Moreover, the transcription observed is resistant to 300 micrograms of alpha-amanitin per ml. These results strongly suggest that prHuTK transcription is under the control of the human rRNA promoter and RNA polymerase I. To further characterize the activity of the human rDNA promoter in vivo, a series of 5' and 3' deletion mutants was tested in this transfection assay. The deletion analysis indicates that a core region of ca. 40 base pairs overlapping the initiation site is critical for transcription. In addition, a region between nucleotides -234 and -131 upstream from the core sequence serves to modulate the efficiency of transcription. Insertion into prHuTK of additional ribosomal nontranscribed spacer DNA or the simian virus 40 enhancer element has no apparent effect on the promoter activity. Surprisingly, RNA polymerase II transcripts synthesized at low levels from two start sites within the core control element of the wild-type RNA polymerase I promoter are activated upon deletion of upstream RNA polymerase I promoter sequences. However, these RNA polymerase II transcripts are not expressed from the endogenous rRNA promoter. PMID- 2983191 TI - Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans. AB - Collagens are the major protein components of the Caenorhabditis elegans cuticle and are encoded by a large family of 40 to 150 closely related but nonidentical genes. We have determined temporal patterns of mRNA accumulation for a large number of collagen genes by screening recombinant phages and plasmids containing cloned collagen genes under high stringency conditions with 32P-labeled cDNA preparations specific for eggs or three postembryonic molts. We find that collagen mRNA levels are regulated both temporally and quantitatively during C. elegans development. Most genes studied exhibit one of four patterns of mRNA accumulation which correlate with changes in cuticle morphology and collagen protein composition during development. Our results suggest that, in general, there is a progressive activation of new collagen genes during normal development. PMID- 2983192 TI - Thymocyte subsets transformed by Abelson murine leukemia virus. AB - The infectious complex of Abelson murine leukemia virus was altered by replacing its usual helper virus, Moloney leukemia virus, with radiation leukemia virus (RadLV). After intrathymic injection of the Abelson-RadLV complex, thymomas arose rapidly, as described previously for injection of the Abelson-Moloney complex. Cell lines were derived from thymomas induced by each Abelson virus complex and were classified according to normal thymus cell phenotypes. Each virus complex induced some cell lines which were like a 0.7% subpopulation of murine thymocytes in that they failed to express the Thy-1 cell-surface antigen. These lines are thus far indistinguishable from some Abelson-derived bone marrow transformants classified as pre-B cells. However, the Abelson-Moloney complex induced some cell lines which expressed low levels of Thy-1 and which shared most markers with immature blast cells of the thymic medulla, whereas the Abelson-RadLV complex induced some lines which were clearly like thymic cortex blast cells. Thus, Abelson virus can induce thymoma cell lines of at least two, and possibly three, distinct phenotypes corresponding to normal thymocyte blast subsets, the determination of which can be influenced by helper virus sequences. PMID- 2983193 TI - Molecular cloning and chromosomal localization of DNA sequences associated with a human DNA repair gene. AB - The genes and gene products involved in the mammalian DNA repair processes have yet to be identified. Toward this end we made use of a number of DNA repair proficient transformants that were generated after transfection of DNA from repair-proficient human cells into a mutant hamster line that is defective in the initial incision step of the excision repair process. In this report, biochemical evidence is presented that demonstrates that these transformants are repair proficient. In addition, we describe the molecular identification and cloning of unique DNA sequences closely associated with the transfected human DNA repair gene and demonstrate the presence of homologous DNA sequences in human cells and in the repair-proficient DNA transformants. The chromosomal location of these sequences was determined by using a panel of rodent-human somatic cell hybrids. Both unique DNA sequences were found to be on human chromosome 19. PMID- 2983194 TI - A vector that replicates as a plasmid and can be efficiently selected in B lymphoblasts transformed by Epstein-Barr virus. AB - Epstein-Barr virus (EBV) transforms human B-lymphocytes into proliferating blasts which are efficiently established into cell lines. The viral DNA in these cell lines is usually present as complete, unintegrated plasmid molecules. A cis acting element of EBV, oriP, permits plasmid maintenance in adherent cells that carry EBV DNA. We constructed a vector, pHEBo, that carries oriP and showed that it is also efficiently maintained as a plasmid when introduced into EBV transformed B-lymphoblasts. The pHEBo vector carries the coding sequences for the hph gene from Escherichia coli such that it can be expressed in mammalian cells and confers resistance to the antibiotic hygromycin B. Hygromycin B kills EBV transformed lymphoblasts at concentrations of 50 to 300 micrograms/ml. The combination of oriP plus the expressed hph gene makes pHEBo useful for the stable introduction of genes on plasmids into EBV-transformed lymphoblasts. Because pHEBo is derived from the plasmid pBR322 it can be easily isolated from lymphoblasts by reintroduction into E. coli. PMID- 2983195 TI - Translocation of oncogene c-sis from chromosome 22 to chromosome 11 in a Ewing sarcoma-derived cell line. AB - Somatic cell hybrids, obtained after fusion of translocation (11;22)-positive Ewing sarcoma cells and Chinese hamster fibroblasts, were assayed for the presence of immunoglobulin C lambda, Philadelphia chromosome breakpoint cluster region, and c-sis oncogene sequences. It was found that c-sis was translocated from chromosome 22 to chromosome 11 in the Ewing sarcoma cells used, indicating that the breakpoint must be proximal to this locus. Moreover, we found that the chromosome 22-linked C lambda and breakpoint cluster region sequences are not translocated. This result confirms an earlier cytogenetic observation that the Ewing sarcoma-associated breakpoint in chromosome 22 is distal to those observed in translocation (8;22)-positive Burkitt lymphoma and in Philadelphia chromosome positive chronic myeloid leukemia. PMID- 2983196 TI - [Morphology of the pituitary gland and adrenal cortex in individuals who died from myocardial infarct]. PMID- 2983197 TI - Solitary cecal ulcer due to cytomegalovirus in a leukemic patient. PMID- 2983198 TI - Gamma-radiation induced ADP-ribosyl transferase activity and mammalian longevity. AB - The levels of ADP-ribosyl transferase (ADPRT) activity following a standardized dose of 100 Gy gamma-radiation have been determined in leukocytes from 12 different mammalian species. Here, we report that the 100 Gy-induced levels of ADPRT correlated in a highly significant manner to the line spans of the mammals. PMID- 2983199 TI - Mutagenesis in UV-irradiated simian virus 40 occurs predominantly at pyrimidine doublets. AB - Phenotypic wild-type revertants from a UV-irradiated temperature-sensitive late mutant (ts BC245) of simian virus 40 (SV40) were isolated after replication in monkey cells at the nonpermissive temperature. The mutations occurring in 7 revertants were identified by DNA sequence analysis of the entire gene involved. All 10 mutations identified constituted single base substitutions, 7 of which occurred opposite pyrimidine doublets. Transitions were 3 times more abundant than transversions. Three base changes did not occur opposite pyrimidine pyrimidine sequences. Exchange of a DNA fragment harbouring the altered base from a revertant with the corresponding fragment from the parental virus, showed that the base substitution was indeed responsible for the reversions to the wild-type phenotype (growth at the restrictive temperature). The data suggest that most base substitutions in highly UV-irradiated simian virus 40 are targeted at sites comprising two adjacent pyrimidines. PMID- 2983200 TI - Interaction between mobile DNA-element-induced lethal mutations and chemical mutagens in the hybrid dysgenic system of Drosophila melanogaster. AB - Using the sex-linked recessive lethal mutation screen, a synergistic interaction is observed for mutations induced by chemical mutagens (ethyl methanesulfonate and dimethylnitrosamine) and the transposable DNA-element system of hybrid dysgenesis in spermatogonial cells of Drosophila melanogaster. Although the mechanism of this interaction is unknown, these results suggest that some chemical mutagens may induce transpositions, hybrid dysgenic cells may be more sensitive to chemically induced genetic damage, or hybrid dysgenesis may inhibit the efficiency of repair of chemically induced lesions. PMID- 2983201 TI - International Commission for Protection Against Environmental Mutagens and Carcinogens. ICPEMC Publication No. 11. The use of recombinant DNA technology to study gene alteration. PMID- 2983202 TI - Antibody to human rotavirus in cow's milk. AB - Rotavirus infection is an important cause of gastroenteritis in infants and young children. Since the virus replicates in the intestinal lumen, we investigated the presence and effectiveness of rotavirus antibody in three forms of milk: raw milk, pasteurized milk, and commercially available infant formulas. Both raw and pasteurized milk contained detectable levels of IgG1 antibody directed at rotavirus. On the other hand, little or no anti-rotavirus antibody was detected in commercially available infant formulas or other sterile milk preparations. The milk samples with rotavirus antibody were capable of inhibiting the replication of simian, bovine, and human rotaviruses in tissue culture. In addition, they were capable of protecting mice from infection and disease in a murine model of rotavirus infection. On the other hand, the formula preparations were incapable of modifying the in vitro replication of rotavirus strains in tissue culture and did not prevent symptomatic gastroenteritis in the mouse model. We conclude that the alteration of milk-processing procedures or the addition of effective antibodies to milk preparations commonly used in the nutrition of young children may alter the clinical course of rotavirus infection or decrease the transmission of rotavirus throughout susceptible populations. PMID- 2983203 TI - Hereditary hypophosphatemic rickets with hypercalciuria. AB - We studied a new hereditary syndrome of hypophosphatemic rickets and hypercalciuria in six affected members of one kindred. In all patients, the manifestations of disease began in early childhood. The characteristic features are rickets, short stature, increased renal phosphate clearance (the ratio between the maximal tubular reabsorption rate for phosphorus and the glomerular filtration rate [TmP/GFR] is 2 to 4 S.D. below the age-related mean), hypercalciuria (8.6 mg of urinary calcium per kilogram of body weight per 24 hours vs. the upper normal value of 4.0), normal serum calcium levels, increased gastrointestinal absorption of calcium and phosphorus, an elevated serum concentration of 1,25-dihydroxyvitamin D (390 +/- 99 pg per milliliter vs. the upper normal value of 110), and suppressed parathyroid function (an immunoreactive parathyroid hormone level of 0.33 +/- 0.1 ng per milliliter and a cyclic AMP level of 1.39 +/- 0.12 nmol per deciliter of glomerular filtrate vs. the lower normal values of 0.3 and 1.5, respectively). Long-term phosphate supplementation as the sole therapy resulted in reversal of all clinical and biochemical abnormalities except the decreased TmP/GFR. We propose that the pivotal defect in this syndrome is a renal phosphate leak resulting in hypophosphatemia with an appropriate elevation of 1,25-dihydroxyvitamin D levels, which causes increased calcium absorption, parathyroid suppression, and hypercalciuria. This syndrome may represent one end of a spectrum of hereditary absorptive hypercalciuria. Our observations support the importance of phosphate as a mediator in controlling 1,25-dihydroxyvitamin D production in human beings. PMID- 2983205 TI - Antibody to lymphadenopathy-associated virus in AIDS. PMID- 2983204 TI - Lymphadenopathy syndrome in two thalassemic patients after LAV contamination by blood transfusion. PMID- 2983206 TI - Classification of lung-cancer histology. PMID- 2983207 TI - Genetic screening for hemophilia A (classic hemophilia) with a polymorphic DNA probe. AB - We have developed a new method of screening for hemophilia A in families at risk for the disease. A DNA probe (St14) that detects a very polymorphic region on the human X chromosome has been shown to be closely linked to hemophilia A. We observed no recombination between the St14 locus and hemophilia A in 12 families studied. The odds in favor of linkage are 4.4 X 10(9) to 1 (lod score, 9.65). The 95 per cent confidence interval for the probability of a recombination between St14 and hemophilia A is 0 to 6.5 per cent. This DNA probe, which is informative in more than 90 per cent of families at risk of hemophilia A, can be used in conjunction with classic biologic assays to identify carriers with an accuracy of 96 per cent or more. If a small risk of misclassification due to crossover between the test and the disease loci is accepted, this DNA marker should allow first-trimester prenatal diagnosis of hemophilia A. Segregation analysis with St14 may thus represent a major improvement in genetic counseling for hemophilia A. PMID- 2983208 TI - Etoposide (VP-16-213). Current status of an active anticancer drug. PMID- 2983210 TI - Extragonadal germ-cell cancer. PMID- 2983209 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 11-1985. A 54-year-old woman with anorexia, nausea, and vomiting and a large mass in the right upper abdominal quadrant. PMID- 2983211 TI - Antibody responses to two Epstein-Barr virus nuclear antigens defined by gene transfer. AB - By transfecting small fragments of Epstein-Barr virus (EBV) DNA into cells, we defined two nuclear antigens, termed M and K, and examined serum from 258 subjects for antibodies against these antigens. We hoped to learn whether such single-antigen systems would clarify the association of EBV with various diseases. Although reactivity to M antigen was found in only 14 per cent of healthy EBV-seropositive subjects, 90 per cent of Chinese and North African patients with nasopharyngeal carcinoma had antibody to M. Nearly all persons (96 per cent) who were EBV seropositive, as judged by their serologic reaction to a nuclear antigen encoded by the complete virus (EBNA), had a reaction to K antigen. However, serum samples from three patients with chronic active EBV infection did not react to K, even though the serum contained anti-M titers above 1:1000. Lymphoid cells from one such patient carried a normal gene for K and made K protein of correct size. Therefore, in this patient the absence of antibody to K had not resulted from a viral mutation that destroyed the K protein. These serologic studies show that some patients with chronic active EBV infection have an abnormal immune response to a specific viral gene product. PMID- 2983213 TI - Anal intercourse as a possible factor in heterosexual transmission of HTLV-III to spouses of hemophiliacs. PMID- 2983212 TI - Diet and 20-year mortality from coronary heart disease. The Ireland-Boston Diet Heart Study. AB - In a prospective epidemiologic study of 1001 middle-aged men, we examined the relation between dietary information collected approximately 20 years ago and subsequent mortality from coronary heart disease. The men were initially enrolled in three cohorts: one of men born and living in Ireland, another of those born in Ireland who had emigrated to Boston, and the third of those born in the Boston area of Irish immigrants. There were no differences in mortality from coronary heart disease among the three cohorts. In within-population analyses, those who died of coronary heart disease had higher Keys (P = 0.06) and modified Hegsted (P = 0.02) dietary scores than did those who did not (a high score indicates a high intake of saturated fatty acids and cholesterol and a relatively low intake of polyunsaturated fatty acids). These associations were significant (P = 0.03 for the Keys and P = 0.04 for the modified Hegsted scores) after adjustment for other risk factors for coronary heart disease. Fiber intake (P = 0.04) and a vegetable foods score, which rose with increased intake of fiber, vegetable protein, and starch (P = 0.02), were lower among those who died from coronary heart disease, though not significantly so after adjustment for other risk factors. A higher Keys score carried an increased risk of coronary heart disease (relative risk, 1.60), and a higher fiber intake carried a decreased risk (relative risk, 0.57). Overall, these results tend to support the hypothesis that diet is related, albeit weakly, to the development of coronary heart disease. PMID- 2983214 TI - Virologic studies in a case of transfusion-associated AIDS. PMID- 2983215 TI - Localization of an insulinoma by ultrasonic endoscopy. PMID- 2983216 TI - AIDS research and "the window of opportunity". PMID- 2983217 TI - HTLV-III, LAV, ARV are variants of same AIDS virus. PMID- 2983218 TI - Evidence for polarization of plasma membrane domains in pancreatic endocrine cells. AB - Polarization of plasma membrane domains is an essential feature of secretory epithelial cells from exocrine glands. The surface of exocrine cells (a typical example is the acinar cell of the pancreas) is separated into an apical domain, where secretion occurs by exocytosis, and a basolateral domain, which senses variations of the internal milieu and is enriched with receptors for various hormones and secretagogues. It is unknown whether secretion is polarized in endocrine cells (except for thyroid follicular cells, which are organized into cavitary structures). To determine whether distinct plasma membrane domains exist in endocrine cells, we infected monolayer cultures of pancreatic endocrine cells with enveloped RNA viruses known to bud selectively from either the apical or basolateral domain in polarized epithelial cells. This asymmetrical budding is thought to reflect the polarized nature of the infected cells, as in non polarized cells such as fibroblasts, the same viruses bud nonselectively from the entire cell surface. We show here that influenza virus and vesicular stomatitis virus (VSV) emerge asymmetrically from cultured pancreatic islet cells; this represents the first evidence for polarization of plasma membrane domains in pancreatic endocrine cells. PMID- 2983219 TI - Glucocorticoid and progesterone receptors bind to the same sites in two hormonally regulated promoters. AB - The glucocorticoid receptor of rat liver recognizes nucleotide sequences near the promoter of mouse mammary tumour virus (MMTV) required for hormonal induction in gene transfer experiments. Similar nucleotide sequences have been found in the human metallothionein gene IIA and in the chicken lysozyme gene, the later induced also by oestrogen, progesterone and androgens. In microinjection experiments, deletion of only 44 base pairs (bp) of the lysozyme promoter (from 208 to -164) results in coordinated loss of progesterone and glucocorticoid dependent gene expression. We show here that purified glucocorticoid receptor from rat liver and progesterone receptor from rabbit uterus yield similar or overlapping exonuclease III footprints in the promoter regions of MMTV and chicken lysozyme. Thus, the regulatory elements for different steroid hormones may be similar or at least share structural features. PMID- 2983220 TI - Duplications of a mutated simian virus 40 enhancer restore its activity. AB - Enhancers are cis-acting control elements which can stimulate at a distance the activity of a variety of eukaryotic promoters. First identified as a repeated 72 base pair (bp) sequence upstream of the simian virus 40 (SV40) early gene promoter, enhancers have since been shown to be associated with numerous other viral and cellular genes. Although there are no strong homologies between the sequences of different enhancers, a number of short and degenerate consensus sequences have been identified, including the 'core' element GTGGA/TA/TA/TG and stretches of alternating purines and pyrimidines which may have the potential to form left-handed Z DNA. To study the functional significance of two alternating purine and pyrimidine sequences in the SV40 enhancer, we have introduced various combinations of point mutations into a modified SV40 enhancer which contained only one copy of the 72 bp element (W.H., Y.G., A. Nordheim and A. Rich, unpublished results); one of these combinations impaired both the activity of the enhancer and growth of SV40. We describe here the structure of 18 revertants of this mutant and suggest that in each of the 18 revertants, the defects of the original mutant have been overcome by simple tandem duplications in the enhancer region, all of which include the 'core' element. PMID- 2983221 TI - Relationship of AIDS to other retroviruses. PMID- 2983222 TI - Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. AB - We have deduced the entire 1,370-amino-acid sequence of the human insulin receptor precursor from a single complementary DNA clone. The precursor starts with a 27-amino-acid signal sequence, followed by the receptor alpha-subunit, a precursor processing enzyme cleavage site, then the beta-subunit containing a single 23-amino-acid transmembrane sequence. There are sequence homologies to human epidermal growth factor receptor and the members of the src family of oncogene products. PMID- 2983223 TI - Plasmid-related anaerobic autotrophy of the novel archaebacterium Sulfolobus ambivalens. AB - Three different species of the genus Sulfolobus, S. acidocaldarius, S. solfataricus (= Caldariella) and S. brierleyi, have been distinguished by the conditions required for optimal growth, by the component patterns of their DNA dependent RNA polymerases and by DNA sequence data. Many isolates of these species are able to grow chemolithoautotrophically using CO2 as the sole carbon source and the oxidation of S(0) with O2 yielding sulphuric acid, as the energy source, though a few others grow only heterotrophically. We show here that a strain of a novel Sulfolobus species, S. ambivalens, is alternatively able to live by an anaerobic mode of chemolithoautotrophy, also using CO2 as the sole carbon source, but using reduction of S(0) with H2, yielding H2S as the energy source. This mode of growth is correlated with the amplification of a plasmid, pSL10. PMID- 2983224 TI - Stable replication of plasmids derived from Epstein-Barr virus in various mammalian cells. AB - Epstein-Barr virus (EBV) infects human B lymphocytes, transforming the infected cells into dividing blasts that can proliferate indefinitely. The viral genome of 172 kilobase pairs (kbp) is a plasmid in most transformed cells. We have identified a region of EBV DNA, termed oriP (nucleotides 7,333-9,109 of strain B95-8), which acts in cis to permit linked DNAs to replicate as plasmids in cells containing EBV DNA. We have postulated the existence of a trans-acting gene allowing oriP function. Here we report that this gene lies in a 2.6-kbp region of the viral genome (nucleotides 107, 567-110, 176) which encodes the EBNA-1 antigen. We show that circular DNAs containing oriP, the EBNA-1 gene and a selectable marker replicate autonomously in cells derived from at least four developmental lineages and from at least three species. We also find that the one third of the EBNA-1 gene repetitive in sequence is not essential for the trans acting function that EBNA-1 gives oriP. PMID- 2983225 TI - Gene deletion and restriction fragment length polymorphisms at the human ornithine transcarbamylase locus. AB - Deficiency of ornithine transcarbamylase (OTC; EC 2.1.3.3), a hepatic mitochondrial enzyme involved in the detoxification of ammonia, is a severe inborn error of metabolism. It is an X-linked disorder which results characteristically in ammonia intoxication, protein intolerance and mental retardation. Early death of affected hemizygous male infants is common, while clinical manifestations in heterozygous females are variable due to random X chromosome inactivation. Prenatal diagnosis by amniocentesis has not been feasible because OTC is not expressed in amniocytes and because no unusual metabolites can be detected in amniotic fluid. Fetal liver biopsy has been performed for some families at risk, but the dangers inherent in this procedure severely limit its usefulness. In this report, we describe the use of a nearly full-length cloned human cDNA to begin to characterize normal and mutant human OTC genes. One of 15 affected males was found to have a partial deletion of the OTC gene. Two distinct restriction fragment length polymorphisms (RFLPs) were identified at the OTC locus using the restriction endonuclease MspI; 69% of women tested were heterozygous for one or both polymorphisms. Identification of these common polymorphisms makes it possible to offer prenatal diagnosis to a large fraction of obligate carriers and to provide information on carrier status to some females at risk. PMID- 2983226 TI - GTP not cyclic GMP enhances secretion from permeabilized platelets. PMID- 2983227 TI - Developmental regulation of T-cell receptor gene expression. AB - In contrast to B cells or their antibody products, T lymphocytes have a dual specificity, for both the eliciting foreign antigen and for polymorphic determinants on cell surface glycoproteins encoded in the major histocompatibility complex (MHC restriction). The recent identification of T-cell receptor glycoproteins as well as the genes encoding T-cell receptor subunits will help to elucidate whether MHC proteins and foreign antigens are recognized by two T-cell receptors or by a single receptor. An important feature of MHC restriction is that it appears to be largely acquired by a differentiating T-cell population under the influence of MHC antigens expressed in the thymus, suggesting that precursor T cells are selected on the basis of their reactivity with MHC determinants expressed in the host thymus. To understand this process of 'thymus education', knowledge of the developmental regulation of T-cell receptor gene expression is necessary. Here we report that whereas messenger RNAs encoding the beta-and gamma-subunits are relatively abundant in immature thymocytes, alpha mRNA levels are very low. Interestingly, whereas alpha mRNA levels increase during further development and beta mRNA levels stay roughly constant, gamma mRNA falls to very low levels in mature T cells, suggesting a role for the gamma gene in T-cell differentiation. PMID- 2983229 TI - New aid to human gene mapping. PMID- 2983228 TI - Structure and transcription of human papillomavirus sequences in cervical carcinoma cells. AB - DNA of human papillomavirus (HPV) types 16 and 18 has been found closely associated with human genital cancer, supporting the concept that members of this virus group are key factors in the aetiology of genital cancer. HPV 18 DNA sequences were also detected in cell lines derived from cervical cancer. We have now analysed these cell lines, HeLa, C4-1 and 756, for the structural organization and transcription of the HPV 18 genome and we find that the HPV 18 DNA is integrated into the cellular genome and is amplified in HeLa and 756 cells. Almost the complete HPV 18 genome seems to be present in 756 cells, with the early region being disrupted into two portions in each integrated copy. In HeLa and C4-1 cells, a 2-3 kilobase (kb) segment of HPV 18-specific sequences is missing from the E2 to L2 region. HPV 18 sequences are specifically transcribed from the E6-E7-E1 region into poly(A)+ RNAs of 1.5-6.5 kb. Hybridization analysis of cDNA clones indicated that some of the transcripts are composed of HPV 18 and cellular sequences. In addition, poly(A)+ RNA hybridizing with HPV 16 DNA was found in two out of three cervical carcinoma biopsies. PMID- 2983230 TI - Identification of a distinct synaptic glutamate receptor on horizontal cells in mudpuppy retina. AB - The separation of ON and OFF channels and the development of an antagonistic surround occur at the first synapse in the vertebrate retina. This functional differentiation is mediated by the action of the photoreceptor neurotransmitter on the ON bipolar, OFF bipolar and horizontal cells, respectively. Glutamate mimics the action of the photoreceptor transmitter on all second-order neurones in fish, amphibian and mammalian retinas. The diversity of cellular responses produced by one neurotransmitter raises the possibility of multiple postsynaptic receptor-ionophore complexes. We reported previously that one glutamate analogue, 2-amino-4-phosphonobutyrate, reveals that the ON bipolar synaptic receptor is pharmacologically different from those of other second-order neurones. The results presented here demonstrate that another glutamate analogue, D-O phosphoserine, selectively antagonizes the synaptic responses of horizontal cells. Taken together, these findings indicate that there are three glutamate like receptor subtypes in the outer retina and suggest a correlation between receptor subtype and the physiological properties of second-order neurones. PMID- 2983231 TI - Co-localization of GABA receptors and benzodiazepine receptors in the brain shown by monoclonal antibodies. AB - The most abundant inhibitory neurotransmitter in the central nervous system, gamma-aminobutyric acid (GABA), exerts its main effects via a GABAA receptor that gates a chloride channel in the subsynaptic membrane. These receptors can contain a modulatory unit, the benzodiazepine receptor, through which ligands of different chemical classes can increase or decrease GABAA receptor function. We have now visualized a GABAA receptor in mammalian brain using monoclonal antibodies. The protein complex recognized by the antibodies contained high- and low-affinity binding sites for GABA as well as binding sites for benzodiazepines, indicative of a GABAA receptor functionally associated with benzodiazepine receptors. As the pattern of brain immunoreactivity corresponds to the autoradiographical distribution of benzodiazepine binding sites, most benzodiazepine receptors seem to be part of GABAA receptors. Two constituent proteins were identified immunologically. Because the monoclonal antibodies cross react with human brain, they provide a means for elucidating those CNS disorders which may be linked to a dysfunction of a GABAA receptor. PMID- 2983233 TI - Investigation of genetic linkage between myosin and actin genes using an interspecific mouse back-cross. AB - The introduction of cloned probes to follow the segregation of DNA restriction fragment length polymorphisms (RFLPs) has led to a revival of mendelian genetics in attempts to map the human genome. In the mouse, however, it has often proved difficult to detect an RFLP with a DNA probe between different inbred strains of the laboratory mouse. To circumvent this problem, we have used two species, Mus musculus domesticus and Mus spretus which interact as sympatric species but can be interbred under laboratory conditions. Because of the relative evolutionary distance between these species, they exhibit polymorphism at many more loci than do different strains of the usual M. m. domesticus laboratory mouse. This is also observed at the DNA level when the sizes of restriction fragments encoding a specific gene are compared. We have used these RFLPs between M. m. domesticus and M. spretus to follow the segregation of genes encoding different isoforms of myosin alkali light chains in the backcross progeny between these species and to compare this with that of other contractile protein genes. No linkage between these genes was observed. PMID- 2983232 TI - A Drosophila genomic sequence with homology to human epidermal growth factor receptor. AB - Vertebrate genomes contain an extensive family of genes possessing varying degrees of homology to the v-src oncogene. Most src-related proteins identified to date are intracellular and membrane-associated, although some are transmembrane proteins and function as receptors for peptide growth factors. Three Drosophila gene sequences related to the v-src gene have been identified, each exhibiting a high degree of homology to one or more of the src-family members encoding an intracellular protein. We have isolated a panel of cloned Drosophila sequences exhibiting weak v-src hybridization and were interested to determine whether any members of this group represented homologues of additional known src-family genes, especially those functioning as growth factor receptors. As we report here, four of these clones, representing overlapping portions of the same genomic segment, hybridized preferentially with the v-erb-B oncogene and were further characterized. The deduced amino-acid sequence from a portion of this Drosophila genomic segment is 77% homologous to the kinase domain of human epidermal growth factor (EGF) receptor, a substantially greater degree of homology than was observed with any other known src-family member. By hybridization with a human EGF receptor complementary DNA clone probe, we demonstrate that the same genomic segment showing homology with the kinase domain also contains sequences related to the extracellular domain of the EGF receptor gene. PMID- 2983234 TI - Reversible cyclic AMP-dependent change in distribution of myosin thick filaments in Dictyostelium. AB - Myosin is thought to act as a major mechanochemical transducer in non-muscle cell motility, but the in situ organization of the molecules has not yet been determined. Here we report the localization of myosin 'rods', analogous to the thick filaments of muscle, by ameliorated immunofluorescence and demonstrate the dynamic translocation of these rods in response to exogenously added cyclic AMP, which is a chemoattractant for Dictyostelium amoebae. On addition of cyclic AMP, we observed instantaneous shedding of the endoplasmic myosin followed by an increase in cortical rods, the original distribution being recovered in a few minutes. We conclude that myosin filaments mediate Dictyostelium cell movement, probably by an assembly/disassembly cycle of the molecules in response to a chemotactic stimulus. PMID- 2983235 TI - Sequence similarity between EGF receptor and alpha 1-acid glycoprotein. PMID- 2983236 TI - Common mononeuropathies. PMID- 2983237 TI - [Initial results of preoperative localization of parathyroid adenomas using subtraction scintigraphy with technetium-99m and thallium-201]. PMID- 2983238 TI - [AIDS and the current significance of serological studies for the presence of antibodies against LAV/HTLV III]. PMID- 2983239 TI - [A seemingly alarming-looking lesion: juvenile giant fibroadenoma of the breast]. PMID- 2983240 TI - [Synaptic mechanisms of the responses of olfactory bulb neurons of the carp to odorants]. AB - Analysis of synaptic responses to some odorants in 16 secondary olfactory neurons and 5 granule cells of carp olfactory bulb revealed that the periodic discharges of secondary neurons, synchronous with the induced waves, were generated on the basis of dendrodendritic inhibitory interaction with granule interneurons. The mechanisms of tonic, asynchronous discharges in some secondary neurons remained not quite clear. PMID- 2983242 TI - [Effect of the toxin of the spider Argiope lobata on neuromuscular transmission]. AB - The action of Argiope lobata (a spider) toxin on the frog neuromuscular junction of the m. sartorius and m. extensor longus digiti IV was investigated. The toxin decreased amplitude of miniature end plate potentials and end plate potentials. The toxin action was reversible. PMID- 2983241 TI - [Synaptic action of vestibular nerve afferents on reticular neurons of the isolated medulla oblongata of the goldfish]. AB - Excitatory post-synaptic potentials (EPSPs) were evoked in reticular neurons of isolated goldfish brian stem by stimulation of the ipsilateral vestibular nerve or by direct intra-axonal activation of single vestibular fibres. Composite EPSPs displayed usually two components. Ca2+-deficient, Mn2+-containing solution abolished the delayed chemical components of EPSPs, but did not affect the short latency responses suggesting dual (electrical-chemical) mode of transmission between some vestibular afferents and reticular neurons. PMID- 2983243 TI - [Mechanisms of the effect of diazepam on the paroxysmal electrical activity of an isolated strip of cat cerebral cortex]. AB - Diazepam effect on paroxysmal electrical activity of an isolated auditory cortex slab and inhibitory responses of its neurons evoked by intracortical electrical stimulation were studied in experiments with cats. Diazepam (2 mg/kg, intravenously) evoked suppression of the paroxysmal electrical activity and increased the number of inhibited neurons both in acute isolated and three-week slabs as compared with the intact cortex. However in this case the number of disynaptic responses decreased, especially in the long-isolated slab. It is supposed that diazepam activated GABA-ergic inhibitory neurons synchronizing inhibition and increasing duration of IPSP. Its action is probably manifested in the first place in the very initial links of cortical neuronal chains. PMID- 2983244 TI - [Effect of an increase in the intracellular concentration of calcium ions on the transmembrane current induced by iontophoretic injection of cAMP into Helix pomatia neurons]. AB - The effect of intracellular Ca2+ concentration increase on transmembrane current induced by cAMP injection into a neuron (cAMP-current) was studied on Helix pomatia neurons under voltage clamp conditions. The elevation of intracellular Ca2+ concentration was produced by iontophoretic injection of Ca2+ into the cytoplasm, generation of action potentials, addition of uncouplers of oxidative phosphorylation into the external solution resulting in Ca2+ efflux from mitochondria. It was found that elevation of the intracellular Ca2+ concentration increases the amplitude of the cAMP-current and in most cases its duration as well. It is suggested that membrane structures responsible for the cAMP-current have two centers of phosphorylation: cAMP-dependent and Ca2+-calmodulin dependent. Possible role of the studied process in integration of signals on the intraneuronal level is discussed. PMID- 2983245 TI - Corticotropin-releasing factor, a new aid in the diagnosis of pituitary-adrenal disorders? PMID- 2983246 TI - Receptors. PMID- 2983247 TI - Nuclear thyroid hormone receptors. PMID- 2983248 TI - Observations on the chemical and physiological properties of urodiolenone, an urinary compound found in hypertension. AB - Urodiolenone is a substance that appears as the glucuronide in the urine of 1 in 3 hypertensive subjects. It is a potent inhibitor of Na+, K+-ATPase in kidney tissue of the guinea pig, as measured by cytochemical assay. Chemical and mass spectrometric evidence is presented, from which it is concluded that urodiolenone is a sesquiterpenoid substance, is a bicyclic enone with a vicinal diol side chain, and has molecular formula C15H24O3. PMID- 2983249 TI - Abnormal erythrocyte pyrimidine nucleotides in uremic subjects. AB - Uremia causes major increases in the erythrocyte (RBC) purine nucleotides, presumably secondary to phosphate retention, but no previous study has been made of the pyrimidine nucleotides, normally absent from RBC. This investigation was prompted by demonstration of the abnormal presence of RBC pyrimidine nucleotides, primarily cytidine triphosphate (CTP) plus cytidine diphosphate-choline (CDP-C) and cytidine diphosphate-ethanolamine (CDP-E), in two types of congenital hemolytic anemia as well as in lead poisoning. These observations suggested an analogy to the RBC membrane dysfunction of uremia. This is a report of the identification of CDP-C and CDP-E as the predominant abnormal pyrimidine nucleotides in the RBC hemolysates of uremic subjects. High-performance liquid chromatography of hemolysates from uremic adults showed a 50% increase in purine nucleotides and the abnormal presence of pyrimidine nucleotides and diesters at approximately 10% of the concentration of the purine nucleotides. By means of UV spectra and 31P nuclear magnetic resonance, these were identified as CDP-C and CDP-E. The increased purine and abnormal pyrimidine nucleotides of uremic RBC were unrelated to the pre- or posthemodialysis state, allopurinol, levels of blood lead, copper and zinc, or RBC pyrimidine 5'-nucleotidase, the cytosolic enzyme that specifically dephosphorylates the pyrimidine nucleotides. Although the accumulation of CTP, CDP-C and CDP-E may be an epiphenomenon of phosphate retention, it also suggests a common pathway to the accelerated hemolysis of chronic renal insufficiency. PMID- 2983250 TI - Effects of propranolol and metoprolol on glucose, cyclic AMP and insulin responses during pharmacologic hyperglucagonemia in hemodialysis patients. AB - Propranolol-induced hypoglycemia in hemodialysis patients has been increasingly recognized. We studied the effects of a nonselective beta-blocker (propranolol) and a beta 1-selective-blocker (metoprolol) on glucose metabolism during pharmacologic hyperglucagonemia in these patients. cAMP and insulin responses to glucagon were noted to be significantly higher in all patients after dialysis. This may possibly be due to the removal of a dialyzable factor suppressing these responses. However, despite a similar cAMP response, patients on propranolol had significantly lower glucose response than those not receiving beta-blocker before and after dialysis. While patients on metoprolol also had impaired glucose response before dialysis, this significantly improved after dialysis possibly due to the removal of active metabolite(s). Results suggest that metoprolol has less interference on glucose response to glucagon than propranolol in hemodialysis. PMID- 2983251 TI - Effect of captopril on blood pressure and renal function in patients with transplant renal artery stenosis. AB - We evaluated 9 patients with transplant renal artery stenosis. Captopril treatment always resulted in a dramatic decrease in renal function. Moreover, only in patients (n = 2) with a stenosis in one out of more renal arteries a satisfying fall in blood pressure was achieved. Surgical reconstruction of the stenosis was successful in 4 out of 5 patients. Angiotensin I-converting enzyme inhibition in transplant renal artery stenosis often leads to loss of graft function, probably due to efferent vasodilation. PMID- 2983252 TI - Hypercalciuric rickets: metabolic studies and pathophysiological considerations. AB - Extensive metabolic studies were performed in a 14-year-old boy suffering from the rare clinical entity known as childhood idiopathic hypercalciuria associated with dwarfism, renal tubular abnormalities and bone lesions. The salient features were: hyperphosphaturia with hypophosphatemia, hypercalciuria with normocalcemia, elevated serum 1,25-dihydroxycholecalciferol[1,25(OH)2D3] levels, marked intestinal hyperabsorption of calcium and phosphorus, with low serum parathyroid hormone (PTH) and urinary adenosine 3':5'-cyclic monophosphate (c-AMP). Bone biopsy confirmed the clinical and radiological diagnosis of rickets. It appears that the following pathophysiological sequence is operating: primary renal phosphate leak with hypophosphatemia, increased 1,25(OH)2D3 synthesis, enhanced intestinal calcium absorption which in turn inhibits release of PTH and c-AMP. Hypercalciuria is seen to be secondary to both avid intestinal calcium absorption and depressed PTH activity, and rickets the result of phosphate depletion. Treatment with oral phosphorus only resulted in an acceleration of growth rate, cure of rickets, and return of urinary calcium excretion to normal values. PMID- 2983253 TI - Effect of the angiotensin converting enzyme inhibitor, captopril (SQ14,225), on orthostatic sodium and water retention in patients with idiopathic edema. AB - Captopril (SQ14,225), an orally administered angiotensin converting enzyme inhibitor, was given to 8 patients with idiopathic edema, in order to study the role of the renin-angiotensin-aldosterone system in orthostatic sodium and water retention. Compared to 5 normal subjects, patients with idiopathic edema showed significantly greater reduction in water and sodium excretion, and greater increment in plasma aldosterone and plasma renin activity, in the upright posture. Captopril significantly restored water and sodium excretion, attenuated the increment in plasma aldosterone, and enhanced the rise in plasma renin activity in patients with idiopathic edema. The effects of captopril on these variables were not remarkable in normal subjects. These results suggest that an enhanced response of the renin-angiotensin-aldosterone system to standing plays an important role in the pathophysiology of idiopathic edema. PMID- 2983254 TI - [Current aspects of pseudomonas meningitis]. AB - The article reports on the incidence, the conditions of occurrence, possibilities and successes of treatment with certain (combinations of) antibiotics, in dealing with cases of pseudomonas meningitis. The various possible substances used for treatment are discussed. Rates of penetration and CSF concentrations of azlocillin and cefsulodin are stated. Alternative possibilities for treatment are pointed out. PMID- 2983255 TI - Intracranial and spinal multiple meningioma appearing after an interval of 22 years. AB - Multiple meningioma is a rare condition which usually occurs in one compartment of the neuraxis. Multiple meningiomas in different neuraxial compartments are an even rarer condition with only a few cases reported in the literature. We describe a case of a 72-year-old woman who developed a parietal meningioma and 22 years later a spinal meningioma. Both tumours were successfully removed. PMID- 2983256 TI - Circadian variations in the inhibition of dopamine release from adult and newborn rat hypothalamus by melatonin. AB - The inhibitory effect of melatonin on evoked dopamine release from the hypothalamus was studied in adult male rats throughout a 24-hour period. The animals were maintained under a daily schedule of 14 h of light (0.00-14.00 h) and 10 h of darkness. The inhibition of dopamine release in vitro by 1 microM melatonin clearly exhibited a 24-hour rhythm with a peak at 5.00 h and almost no inhibition at 15.00 h. The concentrations of melatonin needed to produce this effect were similar throughout the 24-hour cycle, although the actual amount of inhibition at any given concentration of melatonin varied. Other indole derivatives, with the exception of 5-methoxy tryptophol, did not affect significantly the release of 3H-dopamine from the male rat hypothalami. The inhibition of dopamine release by melatonin was not observed in newborn rats but developed during the first week of life, reaching a plateau level between 6 and 7 days postnatally. However, the difference between the amount of inhibition by melatonin at 5.00 h and at 8.00 h existed from the time the inhibition was first observed. The change in amplitude of this difference was due not to differences in the apparent affinity towards melatonin but to increase in the maximal inhibition observed at 5.00 h. The data indicate that the hypothalamic sensitivity to melatonin exhibits a circadian rhythm, and that this develops postnatally prior to the development of circadian variations in melatonin levels, i.e. the 'melatonin rhythm'.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983257 TI - Effects of acute behavioral stress on plasma and cerebrospinal fluid ACTH and beta-endorphin in rhesus monkeys. AB - To elucidate the effect of acute behavioral stress on plasma and cerebrospinal fluid (CSF) concentrations of adrenocorticotropic hormone (ACTH) and beta endorphin, rhesus monkeys were subjected to 30 min of confinement stress. Simultaneous plasma and CSF samples revealed no significant change in CSF ACTH or beta-endorphin up to 120 min after the onset of the stress despite significant elevations in plasma cortisol, ACTH, and beta-endorphin. It is suggested that acute behavioral stress does not alter CSF ACTH or beta-endorphin, and that this information may be clinically useful for future human studies of CSF ACTH and beta-endorphin in neuropsychiatric illnesses. PMID- 2983258 TI - Heterogeneity of congenital motor and sensory neuropathies. AB - Six children suffering from a congenital motor and sensory neuropathy (CMSN) are described. Severe muscle hypotonia, areflexia and a delay of motor development are detectable in all of them. Sural nerve biopsies exhibited an almost complete absence of myelinated fibres and a correspondingly slow nerve conduction velocity (NCV) of less than 10 m/s was detectable in four patients. A few segments with hypermyelination adjacent to gross hypomyelination were seen in the fifth patient, and the NCV was 15 m/s. The sural nerve of the sixth patient showed a loss of thick myelinated nerve fibres, and his NCV was 25 m/s. These results demonstrate the histological heterogeneity of CMSN which was already detected by the NCV. The relation of our findings to the classification of HMSN by Dyck and Lambert (1968) is discussed. PMID- 2983259 TI - Effect of thyroid hormone and serum on the development of Na+, K+-adenosine triphosphatase and associated ion fluxes in cultures from rat brain. AB - The effect of culture conditions, serum supplementation or chemically defined medium and the influence of thyroid hormone were studied on the development of the Na+, K+-adenosine triphosphatase (Na+,K+-ATPase) and on the intracellular content of K+ and Na+ ions in cultures which either were greatly enriched in a neuronal cell type, the cerebellar granule cells, or contained a mixed population of cells (brain reaggregates). Foetal rat brain reaggregates displayed lower Na+,K+-ATPase activity when cultured in chemically defined medium than in the presence of serum. Supplementation of the serum-free medium with thyroid hormone resulted in a rise in the Na+,K+-ATPase activity and [3H]ouabain binding to levels similar to those found in the cultures grown in the serum-containing medium. Thyroid hormone had no significant effect on the Mg2+-ATPase activity and on the intracellular content of Na+ and K+ ions. In the granule cell-enriched cerebellar surface cultures the Na+,K+-ATPase activity was lower when the cells were grown in chemically defined medium compared with the serum-containing medium, and the intracellular Na+ to K+ ratio was higher. Thyroid hormone had no effect on the Na+,K+-ATPase activity, [3H]ouabain binding or Mg2+-ATPase activity. The hormone also failed to influence ATPase activities in cerebellar astrocytes maintained in chemically defined medium. Although thyroid hormone had no effect on the Na+,K+-ATPase activity of cultured cerebellar granule cells, treatment with the hormone resulted in a decrease in the ratio of intracellular Na+ to K+ ion content. The effect of the hormone on the Na+,K+-pump activity in live cells was therefore tested by estimating ouabain-sensitive 86Rb uptake. This was regulated as in other cell types, by the rate of Na+ entry: the Na+-ionophore monensin trebled the rate of 86Rb uptake, which was also increased (+30-100%) by 10% foetal calf serum, the maximal response being obtained by about 20 min exposure to serum. The effect was completely blocked by the Na+/H+ exchange inhibitor amiloride. The factor(s) in the serum responsible for the regulation of the Na+,K+-pump were, however, not the thyroid hormones, which failed to affect 86Rb uptake. On the basis of comparing thyroid hormone effects on the different cultures studied it was concluded that not every type of neural cell is target of the hormone action during development. PMID- 2983260 TI - Autoradiographic localization of thyrotropin-releasing hormone (TRH) receptors in human spinal cord. AB - Thyrotropin-releasing hormone (TRH) exerts many effects upon spinal cord function in animals, and may also play a role in human spinal cord function. We have used the technique of quantitative autoradiography to anatomically localize specific receptors for TRH within human spinal cord. Highest concentrations of TRH receptors were localized within lamina II, the substantia gelatinosa. A moderate density of TRH receptors was found in lamina IX, the motor neurons of the anterior horn. Low levels of TRH receptors were noted throughout the remainder of the gray matter of the human spinal cord, and no TRH receptors were localized within white matter. This anatomic distribution of TRH receptors within the human spinal cord is consistent with the localization of endogenous TRH and the effects of exogenously applied TRH in animal studies. These results suggest that any effects of TRH on human spinal cord function may be mediated by TRH receptors. PMID- 2983261 TI - Brainstem auditory evoked potentials in chronic renal failure and maintenance hemodialysis. AB - Brainstem auditory evoked potentials (BAEPs) were recorded in 36 chronic uremic patients maintained on hemodialysis and in 10 normal controls. Both absolute response latencies and central transmission times were affected. There were transitory increases in interpeak latencies of I-II and III-V that were reversed by dialysis. A permanent increase in wave III-V interpeak latency was not reversed by dialysis. Wave II-III interpeak latency was unaffected. PMID- 2983262 TI - Epstein-Barr nuclear antigen and viral capsid antigen antibody titers in multiple sclerosis. AB - To characterize the antibody response to the Epstein-Barr virus (EBV) in MS, we studied serum anti-EBV nuclear antigen (anti-EBNA) and anti-EBV capsid antigen (anti-EBVCA) titers. Both titers were assayed in 93 age- and sex-matched pairs of MS patients and controls. Anti-EBVCA titers were measured by indirect immunofluorescence and anti-EBNA titers by anticomplement immunofluorescence. The seropositivity rate of both anti-EBVCA and anti-EBNA in MS patients was 100%, compared with 84% in controls (p less than 0.0001). Both anti-EBVCA and anti-EBNA titers were significantly higher in MS patients than in controls (p less than 0.0001). The data suggest that EBV has a significant seroepidemiologic association with MS, but they do not define what role EBV antibodies play in the pathogenesis of the disease. PMID- 2983263 TI - An agent orange: case history. PMID- 2983264 TI - Primary small bowel adenocarcinoma presenting as a malignant ovarian neoplasm. PMID- 2983265 TI - Lithium slows neuronal calcium regulation in the snail Helix pomatia. AB - Steady-state and transient changes in intracellular calcium concentrations ([Ca2+]i) of snail neurons (Helix pomatia) were measured by the Ca2+ indicator Arsenazo(III) following manipulation of the extracellular concentration of lithium chloride (LiCl). Application of LiCl in concentrations equivalent to those used in the treatment of manic-depressive illness produces slowing in Ca2+ reequilibration after Ca2+-influx during depolarization, concomitantly with steady-state elevation of [Ca2+]i of about 100 nM, suggesting a change in Ca2+ reequilibration as a prominent action of LiCl. This mechanism may be relevant to the therapeutic effects of LiCl. PMID- 2983266 TI - Selective blockade of an excitatory synapse in rat cerebral cortex by the sigma opiate cyclazocine: an intracellular, in vitro study. AB - In isolated slices of rat cortex, intracellularly recorded responses of pyramidal neurones to N-methylaspartate (NMA) and glutamate and responses to stimulation of the underlying white matter were compared and challenged with the sigma opiate cyclazocine. Cyclazocine blocked responses to iontophoretically applied NMA and one component of the postsynaptic response to stimulation, but left other postsynaptic events and responses to glutamate intact. These results are consistent with the view that sigma opiates act as selective NMA antagonists and block synaptic events mediated by NMA receptors. PMID- 2983267 TI - Blockade of intrathecal 5-hydroxytryptamine-induced antinociception in rats by noradrenaline depletion. AB - The analgesic effect of 5-hydroxytryptamine (5-HT, 200 and 250 micrograms/kg) administered into the lumbar intrathecal space of rats was blocked in the tail flick, hot-plate and shock titration tests of nociception when the animals had previously been treated with N-2-chloroethyl-N-ethyl-2-bromobenzylamine (DSP4), which depletes noradrenaline in the spinal cord. These results suggest an important modulatory role of noradrenaline upon spinal 5-HT mechanisms. PMID- 2983268 TI - Monoclonal antibody against L-histidine decarboxylase for localization of histaminergic cells. AB - The immunochemical and immunohistochemical properties of a monoclonal antibody (Mab HI 113-12) developed in mice against a partially purified preparation of rat gastric L-histidine decarboxylase (HD) were studied. The Mab recognised the HD activity from the antigen and from crude tissue extracts with a high histamine (HA)-synthesizing capacity (stomach, hypothalamus, striatum, mastocytoma). In contrast, neither a bacterial HD nor other decarboxylases (glutamic and DOPA decarboxylases) were immunoprecipitated. In preliminary immunohistochemical studies, staining of the cytoplasm of mastocytoma as well as of hypothalamic neurons, particularly magnocellular ones in the mamillary region, were observed. The latter presumably correspond to the cells of origin of a long ascending histaminergic pathway. PMID- 2983269 TI - A 39-year-old man with thrombocytopenia, cough, fever, and shaking chills. PMID- 2983270 TI - Renal calcium conservation in recurrent stone-formers with idiopathic hypercalciuria. AB - Urinary excretions of calcium and cyclic AMP were studied in male recurrent stone formers after an overnight fast and following an oral calcium load. The results from eight patients with established hypercalciuria (greater than 7.5 mmol Ca/24 h) were compared with those from eight age matched normocalciuric stone-formers. The urinary calcium/creatinine ratio was higher in the hypercalciuric group both when fasting and calcium loaded whilst their urinary cyclic AMP was lower in both 24-h and calcium-loaded collections. Five of the eight hypercalciuric patients exhibited an increased urinary calcium/creatinine ratio whilst fasting. These findings support the view that renal calcium wasting, in association with suppression of parathyroid activity, is common among men with idiopathic hypercalciuria. Dietary calcium restriction may lead to bone loss in patients with obligatory renal calcium wasting and enteric adsorption is rarely applicable to the treatment of stone disease. Therefore, the demonstration of a high fasting urinary calcium/creatinine ratio is a strong indication for therapeutic agents which act to suppress renal calcium loss to treat hypercalciuria. PMID- 2983271 TI - Hepatitis B viral infections: clinical, pathological, serological features and treatment. PMID- 2983272 TI - An analysis of abnormal cervical smears at Dunedin Hospital 1963-82. AB - Analysis of the abnormal cervical smears seen in the Dunedin Hospital during the 20 year period 1963-82 showed a significant increase in abnormal smears in patients 34 years of age and younger during 1978-82. There were also a number of abnormal smears seen in teenagers; indicating early onset of sexual activity. The increase in abnormal smears is expected to continue; especially in the younger subjects and a one yearly screening programme for all sexually active females is advocated. The possible role of the human papilloma virus in cervical neoplasia is discussed. PMID- 2983273 TI - The hormonal effects of pituitary surgery and irradiation: a review of 59 cases. AB - The endocrine function of 59 patients with pituitary tumours who were treated in our unit between 1962 and 1982 is reviewed. There were 20 patients with growth hormone (GH) secreting adenomas, 17 with prolactinomas (1 tumour secreted both GH and prolactin), 14 corticotrophin (ACTH) secreting adenomas and nine non functioning pituitary adenomas. Patients were treated with pituitary surgery and/or irradiation, with and without the addition of drugs. Patients treated with drugs alone were not included. Following combined surgery, radiotherapy and bromocriptine therapy only two acromegalic patients had persistently raised growth hormone levels and five continued to be hyperprolactinaemic. Pituitary irradiation alone was associated with the development of hypothyroidism in three of eight patients. A delayed but significant fall in free thyroxine index (p less than 0.01) was noted in patients receiving 4500 cGy radiotherapy and surgery. Four patients also developed delayed ACTH deficiency at times varying from one to six years after treatment. It is concluded that patients who have received pituitary surgery or irradiation require regular long-term endocrine assessment. PMID- 2983274 TI - Colposcopy in women with papillomavirus lesions of the uterine cervix. AB - Colposcopic examinations for human papillomavirus lesions were performed in 271 women, some of whom had developed concomitant cervical intraepithelial neoplasia since 1981. The colposcopic appearance was classified into one of the following categories: normal, punctate, mosaic, warty, leukoplakial, or combination, and was related to findings in Papanicolaou smears and punch biopsy specimens. There was a good correlation between the colposcopic appearance and the findings in the Papanicolaou smears and punch biopsy specimens, facilitating the diagnosis of the lesions. The accuracy of colposcopy in disclosing the atypias varied according to the growth pattern of the papillomavirus lesions, with the most accurate (100%) in cases of papillomatous condylomas, and the least accurate (50%) in the inverted lesions. White epithelium and combination patterns were most frequently associated with the papillomavirus lesions and cervical intraepithelial neoplasia, as evidenced by both cytology and biopsy. During the follow-up, normal colposcopic appearance increased from 32 to 50%, reflecting the established spontaneous regression of a certain percentage of the cervical lesions, or their regression as a result of biopsy. The results are discussed in terms of the mutually complementary roles of colposcopy, cytology, and biopsy, and in view of the clinical behavior of cervical papillomavirus lesions. Colposcopy is mandatory for adequate prospective follow-up of these patients but should not replace cytology and punch biopsy. PMID- 2983275 TI - Fetal atrial flutter and X-linked dominant vitamin D-resistant rickets. AB - A pregnancy, complicated by hypophosphatemic familial rickets (vitamin D resistant rickets) of a female fetus, associated with atrial flutter and congestive heart failure is presented. Upon review of the literature, only 22 cases of fetal atrial flutter have been reported. The association between hypophosphatemic familial rickets and atrial flutter has not yet been described. PMID- 2983276 TI - Isolated adrenocorticotropic hormone deficiency with antepartum pituitary infarction in a type I diabetic. AB - Isolated adrenocorticotropic hormone (ACTH) deficiency was documented after cesarean delivery in a patient with type I diabetes. A preceding severe headache, subsequent declining insulin requirements, and hypotension are consistent with pituitary infarction in a patient without computed tomography evidence of a pituitary tumor. Antepartum incomplete pituitary infarction with isolated ACTH deficiency is an unusual but potentially catastrophic complication of the pregnant diabetic. PMID- 2983277 TI - Krukenberg tumor of the ovary from an occult appendiceal primary: case report and literature review. AB - A patient with an occult primary adenocarcinoid tumor of the appendix metastatic to the ovary is presented. Only one similar case has been reported previously. A review of the literature of Krukenberg tumors and appendiceal epithelial tumors emphasizes terminology, sources of primary malignancy, treatment, and prognosis. PMID- 2983278 TI - [Role of follicular cells in the inertia period of oocyte maturation in the common frog]. AB - Out of the breeding season the in vitro maturation of Rana temporaria oocytes in the state of maturation inertia or close to it depends on the follicular cells. In 28 females the presence of follicular cells stimulated oocyte maturation, in 12 females inhibited it. Dibutyrylcyclic AMP (5 X 10(-5) M) increased the percentage of maturation of follicle--enclosed oocytes close to the state of maturation inertia; estrone (4 X 10(-5)-10(-7) M and more) decreased the percentage of maturation of oocytes in the state of inertia, both with and without follicular envelopes. PMID- 2983279 TI - A clinicopathologic study of primary adenoid cystic carcinoma of the lacrimal gland. AB - A retrospective study of 26 patients with the diagnosis of primary adenoid cystic carcinoma of the lacrimal gland was performed with use of randomly selected tissue from each tumor. An attempt was made to determine any associations among clinical presentation, survival, and tumor histologic patterns. We found that lower tumor grades were associated with a predominantly Swiss-cheese (cribriform) pattern. Both lower tumor grade and Swiss-cheese pattern were associated with longer survival. The basaloid (solid) pattern or bone invasion was not associated with a shorter survival. Between men and women, there were no differences in clinical presentation, histologic patterns, or survival. Survival was not affected by surgical or radiation therapy. PMID- 2983280 TI - Melanotic neuroectodermal tumor of infancy (retinal anlage tumor). AB - Six cases of the melanotic neuroectodermal tumor of infancy, including light and electron microscopic findings, are presented. The tumor, now believed to originate from neural crest, is usually cured by wide excision; however, malignant cases have been reported. Histopathologically, this lesion resembles neural retina and retinal pigment epithelium. It may involve the orbit but most frequently occurs in the maxilla. One case presented involved the orbit, another the anterior fontanelle, and four presented in the maxilla. PMID- 2983281 TI - Butorphanol and nalbuphine: a pharmacologic comparison. AB - The agonist/antagonist analgesics, butorphanol (Stadol) and nalbuphine (Nubain), are being increasingly employed as intravenous sedation agents; nalbuphine will be available in the future as an oral analgesic. The drugs possess numerous pharmacologic similarities and some dissimilarities. Both are equianalgesic (and nalbuphine is equipotent) with morphine parenterally and codeine orally. Their pharmacokinetics are similar; nalbuphine has a longer duration of action. Both may precipitate an abstinence syndrome in narcotic-dependent persons and will probably be associated with low-level drug abuse potential. They are both agonists of the kappa opioid receptor and partial agonists of the mu receptor. Butorphanol is a partial agonist of the sigma receptor responsible for psychotomimetic effects. The incidence of adverse effects is low, sedation being the most common. In cardiac-risk patients, nalbuphine does not increase cardiac work or oxygen requirements; nor do increasing doses of nalbuphine increase the duration of respiratory depression. Both drugs possess plateau respiratory depressant actions. PMID- 2983282 TI - Effect of antimicrobial mouth rinses on the incidence of localized alveolitis and infection following mandibular third molar oral surgery. AB - Mandibular third molars were removed from 400 patients who were divided into four different preoperative and postoperative rinsing groups using either normal saline solution, chloramine-T, povidone iodine, or sodium bicarbonate. The overall incidence of localized alveolitis and infection was 4.5% and 1.7%, respectfully. The incidence of the two postoperative problems proved not to be statistically significant when the four groups were compared with each other. There seems to be no apparent advantage to the preoperative and postoperative use of an antimicrobial mouth rinse in place of normal saline solution in reducing the incidence of either localized alveolitis or postoperative infections. PMID- 2983283 TI - Adenoid cystic carcinoma of the upper lip. AB - An elderly man had a large adenoid cystic carcinoma (ACC) of the upper lip. Because of the patient's desire not to have a major surgical resection, the primary modality of treatment was irradiation. A local recurrence following radiation therapy did require surgery, however without morbidity. During the postoperative course extreme physical deterioration of the patient necessitated a complete medical work-up. This revealed lung nodules, not seen previously, which were suspected of being malignant. A barium enema also revealed a sigmoid lesion suggestive of malignancy. Since none of the suspected distant sites were histologically examined, the course of the ACC is left to conjecture. Also, it is left to conjecture as to what course might have followed had the patient been treated initially with a surgical resection. The patient died 20 months after the original diagnosis of the lip tumor was made. PMID- 2983284 TI - Computerized tomography of combined carcinomas arising in pleomorphic adenoma. AB - A case of carcinoma ex pleomorphic adenoma of the palate and maxillary sinus is reported. Unlike most such cases, there were two distinct malignant components: adenoid cystic carcinoma and adenocarcinoma. The computerized tomography of this lesion is described. PMID- 2983285 TI - [Adenoid cystic carcinoma of the trachea]. PMID- 2983286 TI - [Effect of beta-mimetics on bronchial mucus transport]. PMID- 2983287 TI - [Leukotrienes and other lipoxygenase derivatives: a group of pharmacologically very active lipid mediators]. PMID- 2983288 TI - [Lipid peroxidation and free radicals. Role in cellular biology and pathology]. AB - Membrane phospholipids attack by oxygen free radicals, i.e. lipidoperoxidation, occurs in normal cellular life. This free radicals attack is controlled by a protective system, both enzymatic (superoxide dismutase, catalase, glutahione peroxidase) and non enzymatic (vitamine E). Imbalance between attack and defense can explain many pathological events. In these events, the oxidation products of unsaturated fatty acids are of fundamental importance, in particular those derived from arachidonic acid (prostaglandins, prostacyclins, thromboxanes and leukotrienes). PMID- 2983289 TI - Testicular torsion in children: scintigraphic assessment. AB - Seventy-five patients with suspected testicular torsion who had scintiscans and adequate clinical follow-up were analyzed retrospectively. The scintiscans diagnosed 12/13 cases of surgically proven missed torsion and 3/3 cases of surgically proven acute complete torsion. The scintiscans successfully distinguished all 35 cases of epididymo-orchitis, 14 cases of torsion of the appendix testes and 8 other miscellaneous conditions from testicular torsion. The incidence of testicular torsion in our patients undergoing scrotal scintigraphy was approximately 24%. PMID- 2983290 TI - Symposium on infections in the compromised host. The acquired immunodeficiency syndrome. AB - The Acquired Immunodeficiency Syndrome (AIDS) is a recently recognized syndrome caused by a newly described retrovirus, Human T-Cell Lymphotropic Virus-III(HTLV III). A disease that selectively attacks the immune system, manifested by opportunistic infections and unusual neoplasms, AIDS has continued to be confined primarily to several unique at-risk populations. AIDS has evoked unprecedented interest in the medical community, and care of patients with this nearly universally fatal syndrome presents many unique challenges to the health care team. PMID- 2983291 TI - An overview of cancer in children in the 1980s. AB - Multidisciplinary teams, therapeutic research, and large successful clinical trials have led to the exciting improved survival outlook in pediatric oncology. The development of sophisticated supportive care measures and the identification of significant prognostic variables within disease categories have dramatically altered the management and outcome for many children with cancer. Prolonged survival has focused attention on the quality of life and strategies to enable these children and their families to cope effectively with chronic, life threatening illness. Research is ongoing on several fronts: to find innovative treatment approaches for children who currently have a poorer prognosis, to minimize or prevent acute and late toxicities by modifying treatment plans so less intensive treatment can be given to patients with a low risk of disease recurrence, and to increase our understanding of the epidemiology and etiology of childhood cancer. With the continued efforts of researchers in the laboratory and at the bedside, prevention of these catastrophic diseases may some day become a reality. PMID- 2983292 TI - [Carcinoembryonic antigen (CEA) in the blood serum of patients with primary bronchial cancer]. PMID- 2983293 TI - Binding characteristics of inhibin in rat ventral prostate. AB - Homogeneous preparation of human seminal plasma inhibin (molecular weight 13,500) was iodinated with 125I to a specific activity of about 40-45 microCi/micrograms and tested for binding to a rat prostate crude membrane preparation. The binding of inhibin was a saturable process as well as time and temperature dependent. This binding was displaceable in a dose-dependent manner by unlabelled inhibin. Other hormones such as LH, FSH, Prl, and TSH from rat or human origin did not influence the binding of labelled inhibin to rat prostate membrane. Of various age groups of rats studied, the maximum binding was observed in 75-day-old rats. The radiolabelled inhibin also showed specific binding to rat pituitary. The preliminary studies regarding involvement of steroids in the control of inhibin receptors in prostate and pituitary indicated that testosterone activates the inhibin receptors at the prostatic level whereas estradiol did not have any effect. However, estradiol increases the pituitary receptors for inhibin as compared to testosterone. PMID- 2983294 TI - Antiprostate carcinoma monoclonal antibody (D83.21) cross reacts with a membrane antigen expressed on cytomegalovirus-transformed human fibroblasts. AB - Virological and epidemiological studies have implicated human cytomegalovirus (HCMV) as a possible etiological agent of prostate cancer. Because of the suspected associations, this laboratory tested the reactivity of a prostate associated monoclonal antibody with HCMV-transformed cells. This mouse monoclonal antibody, D83.21, reacts with a membrane antigen on prostate and bladder tumor cells and does not bind to a variety of other malignant or normal cells. The results of this study indicated that the prostate-associated antibody bound to a membrane antigen on HCMV-transformed cells as detected by radioimmunoassay, immunofluorescence, and complemented-dependent cytotoxicity. This cross reactivity appeared to be specific for HCMV-transformed cells and did not react with HCMV-infected cells or those transformed by other viruses. Antibody affinity chromatography, used to isolate the D83.21-reactive protein, revealed two peptides of 60 and 28 kd on both prostate tumor and HCMV-transformed cells. The results suggest that D83.21 reacts with a common cell surface protein expressed on HCMV-transformed cells and urogenital tumors. PMID- 2983295 TI - Nutritional supplementation with megadoses of vitamin B6. Effective therapy, placebo, or potentiator of neuropathy? PMID- 2983296 TI - The effect of tap water, carbonated water, sodium bicarbonate, and calcium chloride on blood acid-base balance in cockerels subjected to heat stress. AB - Twenty Hubbard cockerels each weighing 2.4 kg were surgically fitted with plastic cannulae in the carotid artery and crop. A solution of either tap water (TW), carbonated water (CW), 2% sodium bicarbonate (HCO3-), or 3.5% calcium chloride (CaCl2) with pH of 7.8, 5.2, 8.0, and 7.4, respectively, was infused into the crop at .41 ml X min-1 X kg body weight (BW)-1. Blood pH, carbon dioxide partial pressure (PCO2), and respiratory rate (RR) were measured at 15-min intervals during a 90-min thermoneutral period (25 C) followed by a 90-min heat stress (37 C). Polypnea occurred in all treatments in the 37 C environment. Infusion of HCO3 increased blood pH throughout the thermoneutral and heat stress periods, and CaCl2 infusion decreased blood pH throughout the experimental periods. At thermoneutral temperature, TW and CW infusions did not affect blood pH or PCO2. During heat stress, blood pH increased in TW and CW treatments; change and final pH were significantly lower (P less than .05) for CW- compared with TW-infused birds. Although blood PCO2 decreased in all treatments with the imposition of heat stress, blood PCO2 was significantly greater (P less than .05) in birds infused with HCO3- at the end of the heat-stress period. The results demonstrate that CW treatment produced a more favorable acid-base balance during acute heat stress than TW by reducing blood pH change during thermal polypnea. Also, high levels of NaHCO3 and CaCl2 intake can produce abnormal acid-base equilibrium conditions during the heat stress-induced respiratory alkalosis. PMID- 2983297 TI - Effects of corticotropin and heat on corticosteroid-binding capacity and serum corticosteroid in White Rock chickens. AB - The effects of the administration of corticotropin (ACTH) or high environmental temperature on serum maximum corticosteroid-binding capacity (MCBC) and serum corticosteroid concentrations were studied in 6- to 8-wk-old White Rock chickens. Highly significant increases in serum corticosteroids produced by either intramuscular or intravenous injections of ACTH were followed by significant reductions in MCBC within 24 hr. A single high-temperature episode (46 C) caused a significant increase in serum corticosteroids when blood was sampled immediately following exposure to heat; however, serum corticosteroids were not significantly increased following two heat episodes spaced 11 hr apart. Exposure to high temperature did not alter MCBC levels. PMID- 2983298 TI - Tests of association of lymphocyte alloantigen genotypes with resistance to viral oncogenesis in chickens. 2. Rous sarcoma and lymphoid leukosis in progeny derived from 6(3) X 15(1) and 100 X 6(3) crosses. AB - Chickens from inbred line RPRL 6(3) are resistant to virally-induced Marek's disease (MD) and lymphoid leukosis (LL) and are relatively strong regressors of virally induced Rous sarcomas. Line 6(3) chickens are homozygous for three independent loci determining surface alloantigens of lymphocytes, Th-1 and Ly-4 on thymus-derived (T) cells and Bu-1 on bursa-derived (B) cells. In contrast, RPRL Lines 100 and 15(1) are highly susceptible to MD and LL and are weaker regressors of Rous sarcomas than 6(3). RPRL Lines 100 and 15(1) are homozygous for different alleles at the Th-1 and Ly-4 loci than 6(3), whereas only Line 100 differs from 6(3) at Bu-1. To test the possible association of these three antigen loci with variations in resistance to virally-induced tumors, homozygous genotypes among later generations derived from 6(3) X 15(1) and 100 X 6(3) crosses were identified by means of immunofluorescence testing of peripheral blood lymphocytes. Test progeny were then bred from matings of homozygous parents. No association was found between Rous sarcoma regression and the Th-1 and Ly-4 T cell alloantigen loci in F6 (6(3) X 15(1)) chickens, but the Bu-1 locus influenced rate of regression in F3 (100 X 6(3)) chickens. The Ly-4 in F6 (6(3) X 15(1)) and Bu-1 in the F3 (100 X 6(3)) chickens did not influence the incidence of LL. However, Th-1 influenced LL in F3 (100 X 6(3)) chickens (P less than .05) and possibly influenced LL in F6 (6(3) X 15(1)) chickens (P less than .09). PMID- 2983299 TI - Iron, transferrin, and acid and alkaline phosphatase in healthy turkeys and in turkeys inoculated with the lymphoproliferative disease virus. AB - Presented are data on iron-binding capacity determinations in the serum of turkeys infected with lymphoproliferative disease (LPD) virus and in healthy males and females (laying eggs and nonlaying) from a breeding flock. Also presented are results of serum and tissue total acid and alkaline phosphatase determinations in turkey poults infected with LPD virus and their uninfected controls and of serum enzyme levels in healthy males and females from the breeding flock. There was no significant alteration in total iron binding capacity (transferrin level) in the serum of turkeys with LPD. Turkey poults inoculated with LPD virus showed a significant decrease in serum alkaline phosphatase activity 4 and 7 weeks postinfection (pi), and a decrease in serum acid phosphatase activity 7 weeks pi. Acid and alkaline phosphatase activity determined in the spleen and pancreas (organs with pronounced tumor involvement) 7 weeks pi did not differ significantly from that of healthy controls, although there was a tendency for both enzymes to decline in the pancreas of the infected turkeys. Healthy laying female turkeys demonstrated marked elevation in serum transferrin level and in acid and alkaline phosphatase activity, as compared with males of the same age. Serum alkaline phosphatase of turkey poults was markedly higher than that of adult turkeys. PMID- 2983300 TI - Simultaneous application of live turkey herpesvirus and infectious bursal disease vaccines against Marek's disease and infectious bursal disease. AB - The efficacy of cell-associated (CA) and cell-free (CF) infectious bursal disease (IBD) vaccines in maternal antibody (MAb)-bearing broiler chickens was compared. Compatibility and potency of a mixed vaccine combining live CA turkey herpesvirus (HVT) and CA IBD vaccine viruses were also tested against gross lesions induced by Marek's disease (MD) or IBD in specific-pathogen-free chickens. Both CA and CF IBD vaccines provided significant protection against gross bursal lesions of IBD in the presence of MAb. Although chickens receiving the CA vaccine showed a slightly higher (60 to 86.7%) protective index (PX) than CF vaccine recipients (40 to 66.7%), there was no statistical difference in their PX between comparable CA and CF IBD vaccines. The HVT and IBD virus (IBDV) were compatible when they were combined as a live bivalent mixed HVT-IBDV vaccine. Based on the incidence of gross lesions induced by challenge exposure to MD or IBD, potency of the mixed vaccine was similar to that of the monovalent HVT or IBD vaccine. Antagonism did not occur between the two component vaccines when they were applied simultaneously. PMID- 2983301 TI - The pain system. The neural basis of nociceptive transmission in the mammalian nervous system. PMID- 2983302 TI - Benign breast disease. PMID- 2983303 TI - [Initial modification of ACE levels by various corticoid doses in patients with sarcoidosis II]. PMID- 2983304 TI - [Caudal epidural anesthesia in children. Study of endocrine changes]. AB - Changes in serum levels of corticotrophin (ACTH), immunoreactive beta-endorphin, antidiuretic hormone and cortisol were compared in children undergoing minor surgery under either general anaesthesia with halothane or epidural anaesthesia by the caudal route. A rapid and major increase in hormone levels was observed under general anaesthesia but not under epidural anaesthesia. PMID- 2983305 TI - [Erythroblastopenia and febrile meningeal syndrome caused by parvovirus in a child homozygotic for sickle cell anemia]. PMID- 2983306 TI - Point mutation generates constitutive expression of an inducible eukaryotic gene. AB - We describe the analysis of two cis-dominant mutations that result in constitutive expression of the inducible CAR1 gene from yeast. One mutation results from insertion of a Ty element just upstream from the point where CAR1 specific transcription begins. The other mutation is a C-to-G transversion at position -153. Isolation of this point mutation, outside of the transcribed region of CAR1, suggests that expression of this gene is regulated at transcription. It also demonstrates the feasibility and usefulness of analyzing the regulatory sequences of eukaryotic genes on a nucleotide-by-nucleotide basis. PMID- 2983307 TI - Characterization of a platelet-activating factor receptor antagonist isolated from haifenteng (Piper futokadsura): specific inhibition of in vitro and in vivo platelet-activating factor-induced effects. AB - Platelet-activating factor (PAF) is a potent lipid mediator of inflammation and asthma. Using a receptor preparation of rabbit platelet membranes, we identified a novel antagonist of PAF in the methylene chloride extract of a Chinese herbal plant, haifenteng (Piper futokadsura). The active antagonist, kadsurenone, was isolated and characterized in several in vitro and in vivo assays. It is a specific and competitive inhibitor of PAF binding to its receptor with a Ki of 5.8 X 10(-8) M vs. a Ki of 6.3 X 10(-9) M for PAF itself. It inhibits PAF-induced aggregation of rabbit platelets and human neutrophils at 2.4-24 microM, without showing any PAF agonistic activity. It potently inhibits PAF-induced degranulation of human neutrophils at 2.5-50 microM, also without any agonist activity. Kadsurenone is active orally at 25-50 mg/kg of body weight in blocking PAF-induced cutaneous permeability in the guinea pig. It also inhibits PAF induced increases of hematocrit and circulating N-acetylglucosaminidase in the rat at greater than 10 mg/kg i.p. in a dose-dependent manner. Kadsurenone does not interfere with the function of several pharmacological mediators and receptors tested. Its structural specificity is evidenced by the poor PAF antagonistic activities of three related structures isolated from the same haifenteng extract. PMID- 2983308 TI - Complete nucleotide sequence of the genome of bovine leukemia virus: its evolutionary relationship to other retroviruses. AB - We report the complete 8714-nucleotide sequence of the integrated bovine leukemia virus genome and deduce the following genomic organization: 5' LTR-gag-pol-env pXBL-3' LTR, where LTR represents a long terminal repeat and pXBL represents a region containing unidentified open reading frames. This genomic structure is similar to that of human T-cell leukemia virus. The LTR contains a putative splice donor site in the R region. The gag gene encodes a precursor protein with the form NH2-p15-p24-p12-COOH. The NH2- and COOH-terminal regions of the pol product show stronger homologies with those of avian, rather than murine, type C retrovirus, and its structure is identical to that of avian virus. The env gene encodes a surface glycoprotein (gp51) and a transmembrane protein (gp30). In contrast to the pol product, the gp30 shows stronger sequence homology with a murine, rather than avian homologue, indicating the chimeric nature of the bovine leukemia virus genome. Comparisons of the best conserved pol sequences and overall genomic organizations between several major oncoviruses allow us to propose that bovine leukemia and human T-cell leukemia viruses constitute a group, designated as type "E," of Oncovirinae. PMID- 2983309 TI - Identification of the components necessary for adenovirus translational control and their utilization in cDNA expression vectors. AB - A transient expression system was used to study the role of the adenovirus late and simian virus 40 (SV40) early mRNA leader sequences and adenovirus virus associated (VA) RNAs in mRNA translation. Hybrid transcription units containing the adenovirus late and SV40 early promoters fused to various coding regions were introduced into monkey COS cells on plasmids containing a SV40 origin of replication. The translational efficiencies of the mRNAs produced from these plasmids were determined after alterations in the viral leader sequences or in the presence of VA RNAs provided by adenovirus infection of the transfected cells or by cotransfection with plasmids containing the VA genes. Efficient translation of mRNA with either adenovirus or SV40 leader sequences is dependent upon the presence of VA RNA. Translational stimulation by VA RNA of mRNAs containing the adenovirus tripartite leader sequences is dramatically reduced if leader exons 2 and 3 are removed or if their orientation is altered. Sequence analysis has indicated a homology between the nontranslated 5' end of SV40 early mRNA and sequences at the border of the 2nd and 3rd tripartite leader exons, which may be responsible for the increased translation of these mRNAs in the presence of VA RNA. PMID- 2983310 TI - The herpes simplex virus amplicon: analyses of cis-acting replication functions. AB - Previous studies have shown that defective virus vectors (amplicons) derived from herpes simplex viruses could be efficiently propagated in virus stocks in the presence of trans-acting helper virus functions. The present study established that two separate cis-acting functions--a DNA replication origin and a cleavage/packaging signal--are required for amplicon propagation. Using deleted derivatives of cloned amplicons, we mapped one of the viral DNA replication origins (ori-2 or oriL) at coordinate 0.422 of the standard HSV-1 genome and at an equivalent position within the HSV-2 genome. PMID- 2983311 TI - The first intron of the human growth hormone gene contains a binding site for glucocorticoid receptor. AB - Glucocorticoid receptor (GCR) protein stimulates transcription from a variety of cellular genes. We show here that GCR partially purified from rat liver binds specifically to a site within the first intron of the human growth hormone (hGH) gene, approximately 100 base pairs downstream from the start of hGH transcription. GCR binding is selectively inhibited by methylation of two short, symmetrically arranged clusters of guanine residues within this site. A cloned synthetic 24-base-pair deoxyoligonucleotide containing the predicted GCR binding sequence interacts specifically with GCR. The hGH binding site shares sequence homology with a GCR binding site upstream from the human metallothionein II gene and a subset of GCR binding sites from mouse mammary tumor virus. All of these binding sites for this eukaryotic transcriptional regulatory protein show remarkable similarity in overall geometry to the binding sites for several prokaryotic transcriptional regulatory proteins. PMID- 2983312 TI - Molecular cloning and complete sequence determination of RNA genome of human rhinovirus type 14. AB - The genomic RNA of human rhinovirus type 14 was cloned in Escherichia coli and the complete nucleotide sequence was determined. The RNA genome is 7212 nucleotides long. A single large open reading frame of 6536 nucleotides was identified, which starts at nucleotide 678 and ends 47 nucleotides from the 3' end of the RNA genome. Comparisons of the specified proteins with those of other picornaviruses showed a striking homology (44-65%) between rhinovirus and poliovirus. The rhinovirus genomic RNA is rich in adenosine (32.1%) and strongly favors an adenosine or uridine in the third position of codons. The predicted map locations of all the rhinovirus structural and non-structural proteins and their proposed proteolytic cleavage sites are described. PMID- 2983313 TI - A synthetic peptide substrate specific for casein kinase II. AB - A synthetic peptide having the sequence Arg-Arg-Arg-Glu-Glu-Thr-Glu-Glu-Glu was found to serve as a convenient substrate for the protein kinase generally referred to as casein kinase II. The enzyme exhibited an apparent Km of 500 microM for the peptide, as compared to an apparent Km of 50 microM for casein. The maximum velocities for phosphorylation of the peptide and of casein were similar. The peptide was not phosphorylated by any of eight other protein kinases, all of which were shown to be active toward their known substrates. The peptide was used to monitor activity during steps in the purification of casein kinase II from bovine liver. These experiments demonstrated that with this peptide it is now possible to obtain specific measurements of casein kinase II activity in crude enzyme preparations. PMID- 2983314 TI - The structure of Marek disease virus DNA: the presence of unique expansion in nonpathogenic viral DNA. AB - DNA of Marek disease virus (MDV) consists of two unique regions UL and US flanked by long inverted repeat regions TRL and IRL, and short inverted repeat regions TRS and IRS, respectively, similar to herpes simplex virus DNA. Comparison of restriction patterns between pathogenic and nonpathogenic MDV DNA was made to identify a region of viral DNA different between these two types of MDV, as it may be responsible for the tumorigenicity of MDV in chickens. The results indicated that BamHI-D and -H, located at the long inverted repeat regions TRL and IRL, were specifically expanded in nonpathogenic viral DNA. The location of the expanded region has been determined within 1.5 kilobase pairs of the Bgl I/Pst I fragment of BamHI-D and -H, close to the junction between the inverted repeat and the unique region. The possibility that a gene responsible for tumor induction may be disrupted by such expansion has been discussed. PMID- 2983315 TI - Transposable element IS1 intrinsically generates target duplications of variable length. AB - Target duplication during transposition is one of the characteristics of mobile genetic elements. IS1, a resident insertion element of Escherichia coli K-12, was known to generate a 9-base-pair target duplication, while an IS1 variant, characterized by a nucleotide substitution in one of its terminal inverted repeats, was reported to duplicate 8 base pairs of its target during cointegration. We have constructed a series of transposons flanked by copies of either the normal or the variant IS1. The analysis of their transposition products revealed that transposons with normal termini as well as those with variant termini can intrinsically generate either 9- or 8-base-pair target duplications. We also observed that a normal IS1 from the host chromosome generated an 8-base-pair repeat. The possible relevance of the observation for the understanding of transposition processes and models to explain the variable length of target duplications are discussed. PMID- 2983317 TI - Molecular cloning of the gene for the RNA-processing enzyme RNase III of Escherichia coli. AB - A ColE1 plasmid from the Clarke and Carbon collection [Clarke, L. & Carbon, J. (1976) Cell 9, 91-99] that contains a 14.4-kilobase Escherichia coli DNA insert complements the rnc-105 mutation, which destroys the activity of the RNA processing enzyme RNase III. This insert and smaller restriction endonuclease fragments derived from it were cloned into the plasmid pBR329. A number of these recombinant plasmids complemented the rnc-105 mutation in a recA genetic background. The smallest cloned fragment that compensated for the rnc-105 mutation was 1.3 kilobase in size. This fragment led to the synthesis of two polypeptides. One of these polypeptides was 25,300 daltons and corresponded in size to the subunit of RNase III. Fragments cloned in opposite orientations led to synthesis of RNase III, indicating that the cloned fragments contained an endogenous promoter. Extracts of an rnc+ E. coli strain containing an rnc+ plasmid had at least 10 times more RNase III activity than did an analogous strain containing the pBR329 plasmid. PMID- 2983316 TI - Complete nucleotide sequences of three VH genes in Caiman, a phylogenetically ancient reptile: evolutionary diversification in coding segments and variation in the structure and organization of recombination elements. AB - Complete nucleotide sequences are described for three caiman (Caiman crocodylus crocodylus) immunoglobulin VH genes (C3, E1, and G4) that hybridize with a murine VH probe. The E1 and G4 genes are physically linked (intergenic distance, approximately equal to 6.5 kilobases) in the same transcriptional orientation but are not directly contiguous with the C3 gene. When the coding segments, including both framework and complementarity-determining regions, of these genes and the murine probe sequences are compared by metric analysis, it is apparent that the caiman genes are only slightly more related to each other than to the mammalian sequence, consistent with significant preservation of nucleotide sequence over an extended period of phylogenetic time. Based on the presence of transcriptionally critical 5' sequences and the absence of terminator codons, frameshift mutations, or other recognizable alterations, the genes do not appear to be pseudogenes. The E1 gene, however, is distinguished from other VH genes because (i) the spacer region within the 3' recombination signal sequence is 12 base pairs, typical of VK genes but not of VH genes, which possess 22- to 23-base-pair spacers and (ii) a near-perfect VH recombination signal sequence is present within the intervening sequence that splits the segment encoding the leader. These studies establish VH gene multiplicity in a species that arose prior to mammalian radiation and provide a description of differences in the configuration and location of recombination elements associated with an otherwise potentially functional gene. PMID- 2983318 TI - Interleukin 2 regulates its own receptors. AB - The cell surface density of high-affinity membrane receptors for the T lymphocytotrophic hormone interleukin 2 (IL-2) determines the rate of T-cell cycle progression. Since 10-fold greater numbers of IL-2 receptor molecules were found by using a radiolabeled monoclonal antibody reactive with IL-2 receptors (anti-Tac) compared with binding of IL-2, the functional relationship of the binding sites recognized by both of these ligands was assessed. In the presence of cycloheximide, IL-2 binding sites declined with a half-time (t1/2) of 2.6 hr, whereas the decay of anti-Tac binding sites was much slower (t 1/2 = 6.4 hr). Moreover, after limited membrane proteolysis, the half-time for the reappearance of IL-2 binding sites was remarkably similar to its decay (t 1/2 = 2.2 hr), while Tac antigen reappearance was markedly retarded, returning to only 20% of original levels within 5 hr after proteolysis. Addition of homogeneous immunoaffinity purified IL-2 to cell populations that expressed equivalent IL-2 and anti-Tac binding sites resulted in a time- and temperature-dependent 8- to 10-fold enhancement of Tac epitope expression and, simultaneously, a 20-30% diminishment of detectable high-affinity IL-2 binding sites. As the magnitude of the IL-2 dependent proliferative response correlated with the density of high-affinity IL 2 binding sites, rather than Tac antigen levels, quantitation of Tac epitope density does not provide a reliable indication of IL-2-responsiveness among activated T-cell populations. Instead, IL-2-receptor interactions actually promote the loss of IL-2 responsiveness by diminishing the density of high affinity binding sites at the time that Tac antigen levels are increased. PMID- 2983319 TI - Association of the N-formyl-Met-Leu-Phe receptor in human neutrophils with a GTP binding protein sensitive to pertussis toxin. AB - Pertussis toxin inhibits the N-formyl-Met-Leu-Phe (fMet-Leu-Phe) mediated human neutrophil functions of enzyme release, superoxide generation, aggregation, and chemotaxis. As pertussis toxin modifies the GTP binding receptor-regulatory protein "Ni," the association of the fMet-Leu-Phe receptor with such a protein was further examined in purified neutrophil plasma membranes. Both fMet-Leu-Phe mediated guanine nucleotide exchange and nucleotide-mediated regulation of the fMet-Leu-Phe receptor are inhibited by pertussis toxin. In addition, membrane pretreatment with pertussis toxin abolishes the fMet-Leu-Phe-mediated inhibition of adenylate cyclase. Actions of pertussis toxin are due to the ADP-ribosylation of a single subunit at 41 kDa in the neutrophil plasma membrane, which comigrates on NaDodSO4 gels with the Ni GTP-binding protein in the platelet plasma membrane. Our results suggest that (i) the fMet-Leu-Phe receptor is associated with a Ni GTP regulatory protein, and (ii) a fMet-Leu-Phe-Ni complex is important in the control of several neutrophil functions, probably involving multiple transduction systems, including adenylate cyclase. PMID- 2983320 TI - Infectious hepatitis B virus from cloned DNA of known nucleotide sequence. AB - The infectivity of cloned hepatitis B viral DNA (HBV) has been tested in chimpanzees to identify a fully functional HBV genome and to assess the risk associated with its handling. Only one of two HBV DNA sequence variants tested was shown to be infectious. "Clone purified" virus of predicted nucleotide sequence was produced from the infectious HBV DNA, and the cloned viral genome was identical in structure with naturally occurring HBV. Infection could be initiated independent of whether circular monomeric or plasmid integrated dimeric forms of the viral genome were inoculated, but the infectivity of the DNA depended on liver cell transfection or intrahepatic injection. Intravenous injection of high doses of infectious HBV DNA did not induce hepatitis, suggesting that there is virtually no risk associated with routine laboratory handling of cloned HBV DNA. PMID- 2983321 TI - Synthetic peptides from four separate regions of the poliovirus type 1 capsid protein VP1 induce neutralizing antibodies. AB - Peptides from different regions of the poliovirus type 1 capsid protein VP1 were synthesized. Antibodies raised against these peptides in rabbits and rats recognized the cognate peptides and denatured VP1. Peptides from four regions of VP1 generated antisera with neutralizing titers specifically against poliovirus type 1. Antisera against all other regions of VP1 failed to neutralize virus infectivity, although some of the antisera clearly bound to native virions. Thus, the neutralizing determinants on VP1 reside in specific noncontiguous regions of the protein and can be defined by specific peptides from these regions. PMID- 2983322 TI - "Transactivation" control signals in the promoter of the herpesvirus thymidine kinase gene. AB - The herpes simplex virus thymidine kinase (TK) gene is transcriptionally activated in trans ("transactivated") by virus-encoded proteins during the infectious cycle. We show that TK plasmids introduced into a HeLa cell transient transcription assay system are also transactivated after infection with a TK- virus. Several aspects of this response are similar to regulation during the normal infectious cycle. Assay of TK promoter deletion and 5- to 10-base-pair substitution mutants in this system reveals that the transactivation response depends on the intactness of 109 base pairs of 5' gene flanking sequence. Differences between these results and analogous assays in the Xenopus oocyte system are discussed. A model for the putative binding of transactivator(s) to the promoter region is presented. PMID- 2983323 TI - Evidence that a 90-kDa phosphoprotein, an associated kinase, and a specific phosphatase are involved in the regulation of Cloudman melanoma cell proliferation by insulin. AB - Proliferation of wild-type Cloudman S91 melanoma cells is inhibited when insulin is included in the culture medium. Using growth inhibition as a selective marker, we isolated variant cell lines that are either resistant to insulin or dependent upon insulin for growth. We have studied the effects of insulin on proliferation by using combined genetic and biochemical approaches. Through a series of genetic hybridization analyses, we have identified three complementation groups and determined that, in general, insulin-sensitivity is dominant to insulin resistance. Through analyses of in vitro protein phosphorylation reactions, we have identified a protein of approximately 90 kDa (pp90) whose phosphorylation is a function of at least one of the complementation groups. Although pp90 is not phosphorylated in extracts of insulin-resistant variants, it is phosphorylated in extracts of insulin-sensitive hybrids formed between complementing resistant variants. Insulin itself exhibits little or no regulation over the phosphorylation of pp90; rather, the ability to phosphorylate pp90 correlates with the ability of cells to respond to insulin. Migration in NaDodSO4/polyacrylamide gels, solubility characteristics, and divalent cation requirements indicate that pp90 is distinct from the 95-kDa beta-subunit of the insulin receptor. Both pp90 and its associated phosphoprotein kinase are found in 30,000 X g pellets of sonicated cell lysates, whereas a specific pp90 phosphoprotein phosphatase activity is found in 30,000 X g supernatant fractions. Phosphorylation of pp90 occurs at tyrosine and serine residues. Our evidence indicates that the state of phosphorylation of pp90 is an important determinant in the regulation of cellular proliferation by insulin. PMID- 2983324 TI - Inhibitors of the cytochrome P-450 enzymes block the secretagogue-induced release of corticotropin in mouse pituitary tumor cells. AB - A mouse pituitary tumor cell line (AtT-20) releases corticotropin (ACTH) in response to a number of secretagogues, including corticotropin-releasing factor (CRF), beta-adrenergic agents, N6,O2'-dibutyryladenosine 3',5'-cyclic monophosphate (Bt2 cAMP), and potassium. The stimulation of ACTH secretion induced by the secretagogues can be blocked by inhibitors of the enzymes that generate (phospholipase A2) and metabolize (lipoxygenase and epoxygenase) arachidonic acid. The phospholipase A2 blockers mepacrine and p bromophenacylbromide inhibited the ACTH release induced by secretagogues. The lipoxygenase inhibitors nordihydroguaieretic acid, butylated hydroxytoluene, and icosatetraynoic acid abolished the ACTH secretion induced by secretagogues, whereas indomethacin, a cycloxygenase inhibitor, did not. Blockers of the cytochrome P-450 epoxygenase, such as SKF 525A and piperonyl butoxide, compounds that have different molecular structures, also suppressed secretagogue-induced ACTH release. These findings suggest that metabolites of arachidonic acid formed via the epoxygenase and/or the lipoxygenase pathway are involved in the stimulation of ACTH release caused by secretagogues. PMID- 2983325 TI - Hormonal response region in the mouse mammary tumor virus long terminal repeat can be dissociated from the proviral promoter and has enhancer properties. AB - The proviral DNA of mouse mammary tumor virus (MMTV) contains a regulatory region closely associated with its promoter, which subjects transcription to the control of glucocorticoid hormones. Delimitation analysis of a chimeric MMTV long terminal repeat-thymidine kinase gene (LTR-tk) has shown that the hormonal regulation sequence is confined to 202 nucleotides preceding the LTR-specific RNA initiation site. A second RNA initiation site (tk-specific mRNA) placed close to the regulatory MMTV sequence by in vitro recombination is also subjected to hormonal stimulation in transfected cells. A series of plasmids with deletions around the LTR cap site progressing from 3' to 5' was made and functionally tested. In vitro deletion of MMTV LTR sequences comprising the RNA initiation sequence and the "TATA" box do not effect hormonal regulation at the tk-specific mRNA start site. Nucleotides up to position -59 from the LTR initiation site could be deleted without influence on the glucocorticoid regulation, whereas deletions to position -65 abolished the hormonal effect on the tk gene transcription. A short MMTV LTR segment containing nucleotides -236 to -52 from the LTR initiation site was recombined with the tk gene or the alpha-globin gene. This fragment confers hormonal inducibility onto the heterologous genes over distances of 0.4 or 1.1 kilobases. The hormonal response region functions when it is placed either 5' or 3' of the regulated gene in both of the possible orientations and is reminiscent of an enhancer sequence. PMID- 2983326 TI - Novel activity of human angiotensin I converting enzyme: release of the NH2- and COOH-terminal tripeptides from the luteinizing hormone-releasing hormone. AB - Angiotensin I converting enzyme (ACE; kininase II; peptidyldipeptide hydrolase, EC 3.4.15.1) cleaves COOH-terminal dipeptides from active peptides containing a free COOH terminus. We investigated the hydrolysis of luteinizing hormone releasing hormone (LH-RH) by homogeneous human ACE. Although this decapeptide is blocked at both the NH2 and COOH termini, it was metabolized to several peptides, which were separated by HPLC and identified by amino acid analysis. A major product was the NH2-terminal tripeptide, less than Glu-His-Trp, and another was LH-RH-(4-10) heptapeptide, indicating that the Trp-Ser bond is cleaved to release the NH2-terminal tripeptide. ACE also released the COOH-terminal tripeptide, Arg Pro-Gly-NH2, and then sequentially the dipeptides Gly-Leu and Ser-Try, leaving less than Glu-His-Trp intact. Thus, less than Glu-His-Trp was formed by both NH2- and COOH-terminal hydrolysis. The cleavage of LH-RH was inhibited by specific ACE inhibitors and by antibody to ACE but not by inhibitors of other enzymes, showing that the hydrolysis was indeed due to ACE. In the absence of chloride, the hydrolysis proceeded at only 16% of the maximal rate (in 500 mM NaCl), but in 10 mM NaCl it increased to 64%. In 500 mM NaCl solution, 86% of the hydrolysis was accounted for by the release of the NH2-terminal tripeptide, whereas in 10 mM NaCl, the COOH-terminal and NH2-terminal cleavage occurred about equally. The Km of LH-RH in 500 mM NaCl was 167 microM and the catalytic constant kcat was 210 min-1. When the NH2-terminal pyroglutamic acid was replaced with glutamic acid ([Glu1]LH-RH), ACE liberated almost exclusively the COOH-terminal tripeptide in 10 mM NaCl. Thus, human ACE, although it is named peptidyl dipeptidase or dipeptidyl carboxypeptidase, can cleave a protected peptide at the NH2 or COOH terminus. The enzyme could be involved in the in vivo metabolism of LH-RH and possibly other blocked peptides. PMID- 2983328 TI - "Retroposon" insertion into the cellular oncogene c-myc in canine transmissible venereal tumor. AB - We examined by Southern blotting the state of the cellular oncogene c-myc in the dog transmissible venereal tumor. The tumor DNA contains a 16.8-kilobase pair (kbp) rearranged c-myc fragment in addition to the normal 15-kbp and 7.5-kbp fragments. We compared the structure of the cloned rearranged c-myc (re-myc) with that of a cloned normal c-myc and found that the rearrangement was due to the insertion of a 1.8-kbp DNA upstream to the first exon of c-myc. The inserted DNA is flanked by 10-base-pair direct repeats and contains a dA-rich tail, suggesting its origin from mRNA. Partial sequence of the inserted element showed 62% homology with the primate interdispersed Kpn I repetitive element. These results provide an example for the behavior of repetitive DNA sequences like the Kpn I family, as movable elements that can transpose nearby to oncogenes or other structural genes and perhaps affect their activity. PMID- 2983327 TI - Bovine papillomavirus contains multiple transforming genes. AB - Bovine papillomavirus type 1 (BPV-1) and its cloned full-length DNA can transform rodent cells in vitro, and the viral DNA persists as an extrachromosomal multicopy plasmid in these transformed cells. Previous studies have identified at least five discrete viral RNAs that are expressed in BPV-1 transformed cells and have shown that these transcripts share a 3' coterminus. To further define the structure of these RNAs and to characterize the functions of individual viral transcripts, we constructed a cDNA library with mRNA from BPV-1-transformed mouse C127 cells using an Okayama and Berg plasmid. From a library of 10(5) independent clones, 200 BPV-1 specific clones were isolated and characterized. Sequence analysis has revealed differential splicing patterns for the mRNA species in BPV 1 transformed cells. In conjunction with the open reading frames (ORFs) deduced from the BPV-1 DNA sequence, it is possible to predict the structure of the potential encoded proteins. The vector used to generate these cDNA clones contains mammalian cell transcriptional regulatory elements, facilitating their functional characterization. We have identified two distinct classes of cDNA clones that can each independently transform mouse C127 cells. One class of cDNA clones contains the E2 ORF intact and the second contains the E6 ORF intact. These two putative viral functions appear to act synergistically in transforming mouse C127 cells in vitro. PMID- 2983329 TI - Determination of the stereostructure of the product of Tn3 resolvase by a general method. AB - A method has been developed for determination of the absolute structure of DNA catenanes. The catenated DNA is partially denatured before being thickened with a coating of RecA protein and spread for electron microscopy. This treatment allows visualization of the orientation of each ring as well as identification of the overlying and underlying DNAs at crossing points. These determinations define the topology of a catenane, providing a powerful means for testing mechanisms of catenane-producing enzymes in DNA recombination and replication. The technique was used to show that the single interlock of the catenated products of site specific recombination mediated by Tn3 resolvase is exclusively of negative sign. The unique topology of the products indicates that resolvase fixes the sum of the number of supercoils between recombination sites at synapsis and the number of such supercoils lost or gained during strand exchange. The data strongly suggest that there are in fact three negative supercoils between synapsed sites; one supercoil is dissolved in the cross-over mechanism, whereas the other two are metamorphosed into the unique catenane interlock. PMID- 2983330 TI - Two genes encoding steroid 21-hydroxylase are located near the genes encoding the fourth component of complement in man. AB - Two genes encoding steroid 21-hydroxylase [21-OHase; steroid 21-monooxygenase; steroid, hydrogen-donor: oxygen oxidoreductase (21-hydroxylating); EC 1.14.99.10], a cytochrome P-450 enzyme, have been located within the HLA major histocompatibility complex. Congenital adrenal hyperplasia due to 21-OHase deficiency is a common inherited disorder of cortisol biosynthesis which is in genetic linkage disequilibrium with certain extended HLA haplotypes. These haplotypes include characteristic serum complement allotypes. A series of cosmid clones was isolated from a human genomic library by using a probe encoding part of the fourth component of complement, C4. These clones also hybridized with a probe encoding most of human 21-OHase. Restriction mapping and hybridization analysis showed that there are two 21-OHase genes, each located near the 3' end of one of the two C4 genes. Hybridization with probes specific for the 5' and 3' ends of the 21-OHase gene showed that the 21-OHase and C4 genes all have the same orientation. The 21-OHase genes 3' to C4A and C4B carry T aq I fragments of 3.2 and 3.7 kilobases (kb), respectively. Both of these fragments are found in genomic DNA of most individuals. In DNA from an individual with the severe, "salt wasting" form of 21-OHase deficiency who was homozygous for HLA A3;Bw47;C4A*1;C4B*Q0(null); DR7, the 3.7-kb Taq I fragment is absent, whereas hormonally normal individuals homozygous for HLA-A1;B8;C4A*Q0;C4B*1;DR3 do not carry the 3.2-kb Taq I fragment. These data suggest that the 21-OHase "B" gene (3.7-kb Taq I fragment) is functional, but the 21-OHase "A" gene (3.2-kb Taq I fragment) is not. PMID- 2983332 TI - Establishment of a line of human fetal glial cells that supports JC virus multiplication. AB - Primary cultures of human fetal brain cells were transfected with plasmid DNA pMK16, containing an origin-defective mutant of simian virus 40 (SV40). Several weeks after DNA treatment, proliferation of glial cells was evident in the culture, allowing passage of the cells at low split ratios. Initially, only 10% of the cells demonstrated nuclear fluorescence staining using a hamster tumor antibody to the SV40 T protein. By the sixth passage, however, 100% of the cells reacted positively to the same antibody. During these early passages, the cells designated SVG began growing very rapidly and acquired a homogeneous morphology. Cell division required only low serum concentrations, was not contact-inhibited, and remained anchorage dependent. These characteristics of the SVG cells have been stable through 25 passages or approximately equal to 80 cell generations. The SV40 T protein is continuously produced in the cells and can direct the replication of DNA inserts in the pSV2 vector, determined by in situ hybridization using biotin-labeled DNA probes, which contains the SV40 replication origin. More importantly, SVG cells support the multiplication of the human papovavirus JCV at levels comparable to primary cultures of human fetal glial cells, producing infectious virus as early as 1 week after viral adsorption. Their brain-cell derivation has been established as astroglial, based on their reactivity with a monoclonal antibody to glial fibrillary acid protein and lack of activity with an anti-galactocerebroside antibody, which identifies oligodendroglial cells. The SVG cells represent a unique line of continuous rapidly growing human fetal astroglial cells that synthesizes a replication proficient SV40 T protein. Their susceptibility to JC virus (JCV) infection obviates a host restriction barrier that limited JCV studies to primary cultures of human fetal brain and thus should allow for more detailed molecular studies of human brain cells and JCV that infects them. PMID- 2983331 TI - Evidence for a role of endogenous corticotropin-releasing factor in cold, ether, immobilization, and traumatic stress. AB - The role of corticotropin-releasing factor (CRF) in four model stresses (cold, ether, immobilization, and trauma) was examined in the guinea pig by using passive immunoneutralization with anti-CRF antiserum. Plasma corticotropin levels were measured at various times after exposure to stress, and groups treated with CRF antiserum were compared with those treated with normal rabbit serum. Of the four stresses tested, ether had the most pronounced effect on corticotropin secretion. Treatment with anti-CRF inhibited most of the ether-induced corticotropin secretory response, the difference between the normal serum- and the anti-CRF antiserum-treated groups being significant at 5 and 10 min (P less than 0.01). Corticotropin responses to cold stress in the two groups differed at the 0.05 level of significance at 10 and 20 min. After administration of trauma (leg fracture), a statistically significant difference (P less than 0.01) between the two groups also was evident, albeit only at 20 min. During immobilization, corticotropin levels differed significantly from control only in the normal serum treated group but not in the anti-CRF-treated group. These findings show that CRF antiserum was effective in reducing corticotropin levels, indicating that CRF has an important role in mediating corticotropin response to stress. The fact that neutralization was incomplete might be due to an inability of the antiserum to sufficiently neutralize the endogenous CRF or, more likely, reflects the contribution of additional mediators, notably catecholamines and vasopressin, of corticotropin release upon stress. PMID- 2983333 TI - Molecular cloning of a cDNA for human alpha-L-fucosidase. AB - A lambda gt11 human hepatoma cDNA expression library was screened with antibodies to human alpha-L-fucosidase, a lysosomal enzyme whose activity is deficient in the human autosomal recessive disease fucosidosis. Three positive clones were identified after screening 9 X 10(6) plaques. One of these was sequenced and found to be spurious, probably representing an out-of-frame cDNA that gave rise to amino acid sequences of unknown length that crossreacted with alpha-L fucosidase. A second clone, lambda AF3, was isolated which, after establishment in Escherichia coli BNN103, gave rise to a fusion protein of Mr 154,000 containing a human fragment of Mr 40,000 that represented 80% of the mature processed enzyme (Mr 50,000). Southern blot analysis of mouse and human chromosomal DNA confirmed the human origin of insert AF3. The nucleotide sequence of AF3 was determined and colinearity was established between 270 nucleotides and 90 amino acids in alpha-L-fucosidase. AF3 was found to contain 1058 base pairs and to code for 347 amino acids of alpha-L-fucosidase. Four potential glycosylation sites were identified. The frequency of lambda AF3 in the hepatoma library was 0.0018%. PMID- 2983334 TI - Precise localization of genes on large animal virus genomes: use of lambda gt11 and monoclonal antibodies to map the gene for a cytomegalovirus protein family. AB - We describe an efficient procedure, which uses monoclonal antibodies directed against specific viral proteins, for the precise mapping of genes on large DNA virus genomes. We have used the technique to locate the gene encoding a family of antigenically related DNA-binding proteins on the 240-kilobase-pair human cytomegalovirus (CMV) genome. A random library of CMV DNA fragments was generated using the prokaryotic vector lambda gt11, which expresses open reading frames as beta-galactosidase fusion proteins in infected Escherichia coli. The library was screened with a mixture of monoclonal antibodies directed against the gene products of interest. The coding sequence for infected cell protein 36 (ICP36) was localized to a 2800-base-pair EcoRI fragment (map coordinates 0.228-0.240) on the CMV(Towne) and CMV(AD169) genomes by using DNA from immunoreactive lambda gt11 as probe. A 5000-nucleotide transcript from this region was detected during the early and late phases of the CMV growth cycle. This transcript directed the synthesis of the predominant member of the ICP36 family when hybrid-selected and translated in vitro. Immunoprecipitation of the in vitro translation product with the same monoclonal antibodies used in the initial mapping confirmed the location of the ICP36 gene. These studies establish the utility of the lambda gt11 expression system for rapid and precise mapping of CMV genes (or other large animal virus genes) that encode proteins for which serological reagents exist. PMID- 2983335 TI - NH2-terminal specificity and axonal localization of adrenocorticotropin binding sites in rat median eminence. AB - Adrenocorticotropin binding sites in the rat median eminence have been localized in vivo. These binding sites occur in the basalar zone, which is rich in axonal endings. Using competitive binding and quantitative light-microscope radioautography, we found that the median-eminence binding site, in contradistinction to the adrenal receptor, binds specifically the residue 4-10 region of the adrenocorticotropin molecule. Using quantitative electron microscope radioautography and median-eminence deafferentation, we localized the binding sites to axon terminals in this region. In time-delayed uptake studies using light-microscope radioautography, we failed to observe concentration of radiolabel in neurons of the medial basal hypothalamus after the direct injection of radioiodinated adrenocorticotropin(1-24) into the median eminence. PMID- 2983336 TI - Lithium induces corticotropin secretion and desensitization in cultured anterior pituitary cells. AB - Lithium stimulated corticotropin (ACTH) secretion by mouse pituitary tumor cells (AtT-20/D16-16) and by normal rat anterior pituitary cells in primary culture. Effects were observed at less than 2 mM LiCl. ACTH secretion was comparable in magnitude to that induced by other secretagogues, was calcium dependent, and was inhibited by somatostatin. Lithium also induced changes in [3H]inositide metabolism; these changes accompanied and were correlated with changes in ACTH secretion. The most prominent and reliable effect was to increase [3H]inositol monophosphate. Other secretagogues had no effect on [3H]inositides in the presence or absence of lithium. Pretreatment with lithium for 3 hr desensitized the cells to the effects of subsequent exposure to lithium. The cells were not desensitized to lithium by pretreatment with other secretagogues, nor were they desensitized by lithium to the effects of corticotropin-releasing factor, high potassium, or forskolin. However, pretreatment with lithium did desensitize the cells to stimulation by phorbol esters. The interaction between lithium and phorbol esters suggests the involvement of inositide metabolism and protein kinase C in the regulation of ACTH secretion and possibly of other hormones or neurotransmitters. It also suggests new avenues of research into the basis of lithium's psychopharmacological effects. PMID- 2983337 TI - An alternative route of infection for viruses: entry by means of the asialoglycoprotein receptor of a Sendai virus mutant lacking its attachment protein. AB - During the first stage of infection, the paramyxovirus Sendai virus attaches to host cells by recognizing specific receptors on the cell surface. Productive virus-cell interactions result in membrane fusion between the viral envelope and the cell surface membrane. It has recently been shown that the ganglioside GD1a and its more complex homologs GT1b and GQ1b are cell surface receptors for Sendai virus. We report in this paper that the temperature-sensitive mutant ts271 of the Enders strain of Sendai virus lacks the viral attachment protein HN and the biological activities of hemagglutination and sialidase activity associated with it when the virus is grown at 38 degrees C. This HN- virus was unable to infect or agglutinate conventional host cells that contained receptor gangliosides and were readily infected by the parental wild-type virus. The HN- virus did, however, attach to and infect Hep G2 cells, a line of hepatoma cells that retains the asialoglycoprotein receptor (ASGP-R) upon continuous culture. This receptor is a mammalian lectin that recognizes galactose- or N-acetylgalactosamine terminated proteins. In accordance with the known properties of this receptor, infection by the HN- virus was abolished by treatment of Hep G2 cells with sialidase, by the presence of Ca2+ chelators, and by competition with N acetylgalactosamine, asialoorosomucoid, and antibody to the receptor. F, the only glycoprotein on the HN- virus, was shown to compete with the galactose-terminated protein asialoorosomucoid for the ASGP-R. The ability of the HN- virus to cause cell-cell fusion of Hep G2 cells indicated that attachment of this virus to the ASGP-R still permitted viral entry by its usual mode--i.e., membrane fusion at the cell surface. These results open up the possibility that enveloped viruses, which contain glycosylated proteins or lipids, may make use of naturally occurring lectins in addition to their normal receptors as a means of attachment to host cells. PMID- 2983338 TI - Transfer RNA is an essential component of the ubiquitin- and ATP-dependent proteolytic system. AB - Protein degradation via the nonlysosomal ATP-dependent pathway in rabbit reticulocytes involves a number of components. In the initial event, ubiquitin, an abundant 76-residue polypeptide, becomes covalently linked to the protein substrate in an ATP-requiring reaction. Once marked in this way, the conjugated protein is proteolyzed in a reaction that also requires ATP. Ubiquitin-marking appears to be important to the progression of cells from one stage to another of the cell cycle; it may also be involved in gene activation. Here we show that tRNA is another essential component of the system. Ribonucleases strongly inhibit the ubiquitin- and ATP-dependent degradation of 125I-labeled bovine serum albumin in the reticulocyte system in vitro. RNAs extracted from fractions of the reticulocyte extract or from mouse cells restore proteolytic activity. When the RNA is fractionated by gel electrophoresis, only the tRNA fraction is active in restoring proteolysis. Furthermore, pure mouse tRNAHis, isolated by immunoprecipitation with patient autoimmune sera, restores the proteolytic activity. The possibility that the level of uncharged tRNA in mammalian cells regulates the ubiquitin- and ATP-dependent proteolytic system is discussed. PMID- 2983339 TI - Stimulation of glucose uptake by transforming growth factor beta: evidence for the requirement of epidermal growth factor-receptor activation. AB - Transforming growth factor beta (TGF-beta), derived from human platelets, stimulates the uptake of 2-deoxy-glucose by cultured cell monolayers 2- to 4 fold. Stimulation can be detected as early as 30 min with as little as 0.1 ng of TGF-beta per ml and maximal effects can be obtained at 2 hr with 1 ng of the growth factor per ml. TGF-beta-induced stimulation of sugar uptake is enhanced by the co-addition of platelet-derived growth factor (10 ng/ml) or epidermal growth factor (EGF, 1 ng/ml). The NR-6 variant of mouse 3T3 cells, which lack EGF receptors, is not stimulated by TGF-beta. Antisera to EGF receptors that block 125I-labeled EGF binding also inhibit TGF-beta stimulation of 2-deoxyglucose uptake, although 125I-labeled TGF-beta binding remains unimpaired. In contrast, anti-sera to the EGF receptor, which do not block EGF binding, have no measurable effect on the TGF-beta-stimulated uptake of 2-deoxyglucose. We confirm that the receptor for TGF-beta is distinct from the receptor for EGF and we conclude that TGF-beta stimulation of 2-deoxyglucose uptake requires the co-activation of the EGF receptor kinase system. PMID- 2983340 TI - Regulated expression of endonuclease EcoRI in Saccharomyces cerevisiae: nuclear entry and biological consequences. AB - In an investigation to determine how proteins are localized within the nucleus of a cell, we demonstrate that the restriction endonuclease EcoRI is able to enter and function within the nucleus of Saccharomyces cerevisiae when this prokaryotic protein is synthesized in vivo. The EcoRI endonuclease was produced in yeast under the transcriptional control of a regulated yeast promoter by ligating a DNA fragment containing only coding sequences for the endonuclease to the promoter element of the yeast GAL1 gene (the structural gene for galactokinase, EC 2.7.1.6). Yeast cells harboring a plasmid containing this promoter-gene fusion are able to grow under conditions that repress transcription from the GAL1 promoter. However, under inducing conditions, these yeast cells are unable to grow. Moreover, rad52 mutants, which are deficient in the repair of double-strand breaks, are more sensitive to the presence of the promoter-gene fusion plasmid than are wild-type cells. We demonstrate that the EcoRI endonuclease activity is present in lysates prepared from yeast transformants grown under conditions that induce transcription of GAL1, but this activity is not detectable in cells grown under conditions that repress transcription from the promoter. Furthermore, analysis of yeast chromosomal DNA shows that the endonuclease enters the yeast nucleus and cleaves DNA specifically at EcoRI recognition sites. PMID- 2983341 TI - An adenovirus vector system used to express polyoma virus tumor antigens. AB - We have used a generalized adenovirus vector system to express the three polyoma tumor (T) antigen proteins under the control of the adenovirus major late promoter. One hybrid virus, Ad-PySVR498, expresses high levels of polyoma middle and small T antigens. A second hybrid virus, Ad-LTSVR545, which contains a cDNA copy of the polyoma A gene, overproduces large T antigen. The T antigens produced are indistinguishable from their authentic polyoma counterparts as determined by immunoprecipitation and partial cleavage by V8 protease. Analysis of polyoma mRNAs encoded by the recombinant viruses showed that they initiate from the adenovirus major late promoter and contain the tripartite leader at their 5' ends. Large T antigen isolated from Ad-LTSVR545-infected cells by immunoaffinity was shown to bind selectively to polyoma DNA sequences that contain the origin of viral DNA replication as well as the sites for transcription initiation. PMID- 2983342 TI - Similarity between the corticotropin (ACTH) receptor and a peptide encoded by an RNA that is complementary to ACTH mRNA. AB - An interesting pattern in the genetic code was recently observed: Codons for hydrophilic and hydrophobic amino acids on one strand of nucleic acid are complemented by codons for hydrophobic and hydrophilic amino acids on the other strand, respectively. The average tendency of codons for "uncharged" (slightly hydrophilic) amino acids is to be complemented by codons for "uncharged" (slightly hydrophilic) amino acids. We have postulated that this pattern can result in the binding of peptides that are encoded by complementary RNA strands and we have presented supporting evidence. In this report we demonstrate the specific and high-affinity binding of naturally occurring peptides [corticotropin (ACTH) and gamma-endorphin] to synthetically derived counterparts that were specified by RNA sequences complementary to the mRNA for ACTH and gamma endorphin, respectively. That this binding might result from one peptide being an "internal image" of the other was strongly suggested by the observation that antibody to the peptide that was encoded by the complementary RNA for ACTH recognized the adrenal cell ACTH receptor. Based on these findings, a theory on the evolution of peptides and their receptors is suggested. PMID- 2983343 TI - The molecular origin of DNA-drug specificity in netropsin and distamycin. AB - X-ray analysis of the complex of netropsin with the B-DNA dodecamer of sequence C G-C-G-A-A-T-T-BrC-G-C-G reveals that the antitumor antibiotic binds within the minor groove by displacing the water molecules of the spine of hydration. Netropsin amide NH furnish hydrogen bonds to bridge DNA adenine N-3 and thymine O 2 atoms occurring on adjacent base pairs and opposite helix strands, exactly as with the spine of hydration. The narrowness of the groove forces the netropsin molecule to sit symmetrically in the center, with its two pyrrole rings slightly non-coplanar so that each ring is parallel to the walls of its respective region of the groove. Drug binding neither unwinds nor elongates the double helix, but it does force open the minor groove by 0.5-2.0 A, and it bends back the helix axis by 8 degrees across the region of attachment. The netropsin molecule has an intrinsic twist that favors insertion into the minor groove of B-DNA, and it is given a small additional twist upon binding. The base specificity that makes netropsin bind preferentially to runs of four or more A X T base pairs is provided not by hydrogen bonding but by close van der Waals contacts between adenine C-2 hydrogens and CH groups on the pyrrole rings of the drug molecule. Substitution of one or more pyrroles by imidazole could permit recognition of G X C base pairs as well, and it could lead to a class of synthetic "lexitropsins," capable of reading any desired short sequence of DNA base pairs. PMID- 2983344 TI - Mapping the location of psoralen crosslinks on RNA by mung bean nuclease sensitivity of RNA.DNA hybrids. AB - An indirect high resolution method has been developed for finding the location of intrastrand crosslinks in RNA. An end-labeled DNA strand that overlaps the approximate crosslink position is hybridized to the RNA and then treated with mung bean nuclease. The resulting digest is analyzed on a sequencing-type gel. The method was tested with the major psoralen crosslink seen in the 16S rRNA of inactivated Escherichia coli 30S ribosomal subunits. This crosslink was previously mapped between residues 930 +/- 25 and a region close to the 3' end by electron microscopy. The new indirect method reveals that the crosslink occurs between residues 919 and 923 and residues 1530 and 1534. When these results are examined in the light of existing consensus secondary structure models for the 16S rRNA, it appears that the Shine-Dalgarno sequence is located close to the peptidyl tRNA binding site. PMID- 2983345 TI - Characterization of the activation of Na+/H+ exchange in lymphocytes by phorbol esters: change in cytoplasmic pH dependence of the antiport. AB - Phorbol 12-myristate 13-acetate and other phorbol esters induce an intracellular alkalinization in rat thymic lymphocytes. An extracellular acidification can be recorded concomitantly. This transmembrane H+ (equivalent) flux is dependent on external Na+ and is amiloride sensitive. Phorbol esters also stimulate an amiloride-sensitive uptake of 22Na+, suggesting activation of Na+o/H+i exchange. Only those phorbol derivatives that are tumor promoters and activators of protein kinase C stimulate the antiport. Activation of the Na+/H+ exchange is brought about by a change in the cytoplasmic pH sensitivity of the antiport. Activation of the Na+/H+ exchanger by phorbol esters results in membrane hyperpolarization, due to indirect stimulation of the electrogenic Na+/K+ pump by the increased intracellular Na+ concentration. Increased Na+/H+ exchange also produces cell swelling, which may be one of the earliest manifestations of the growth-promoting properties of the phorbol esters. PMID- 2983346 TI - Loss of polymorphic restriction fragments in malignant melanoma: implications for tumor heterogeneity. AB - Loss of genetic material at certain chromosomal sites is implicated in the etiology of retinoblastoma and Wilms tumor. Whether specific chromosomal deletions are associated with other types of human cancer needs to be explored. We have examined 24 melanoma cell lines, derived from 21 patients with nonfamilial malignant melanoma, for evidence of somatically induced hemizygosity or homozygosity. Twelve DNA probes, recognizing single-copy restriction fragment length polymorphisms (RFLP) determined by loci on 11 different chromosomes, were used to screen autologous combinations of melanoma cells and either B cells or fibroblasts. Loss of heterozygosity in melanoma cells was identified at 27 of 100 informative loci. These losses occurred at loci on 8 different chromosomes, and the frequency of loss at individual loci varied between 8% and 67%. We conclude that somatic mutations resulting in homozygosity or hemizygosity are common in melanoma and evidently not restricted to specific chromosomes. PMID- 2983348 TI - Characterization of a Q subregion gene in the murine major histocompatibility complex. AB - We have used restriction enzyme digests, Southern blot analysis, and gene transfer experiments to identify a class I gene in the Q subregion of the murine major histocompatibility complex. By comparisons of class I genes from Q congeneic strains, five restriction fragment length polymorphisms were identified. Further studies of mutant (Qa-2-) and wild-type (Qa-2+) BALB/c sublines indicated that at least part of the structural or regulatory gene controlling a Q subregion antigen resides on a 3.7-kilobase Xba I DNA fragment and is absent in all tested Qa-2- strains. The spontaneously occurring Qa-2- BALB/cBy mutant appears to have an extensive deletion in this region. The identity of this gene was confirmed by gene transfer experiments as well as by the use of a single-copy probe. PMID- 2983347 TI - gp140, the C3d receptor of human B lymphocytes, is also the Epstein-Barr virus receptor. AB - The relationship between gp140, the membrane C3d receptor (CR2) of human B lymphocytes, and the Epstein-Barr virus receptor (EBVR) was analyzed by using the polyclonal anti-gp140, previously prepared by immunizing rabbits with highly purified gp140 (isolated by some of us) from CR2/EBVR-positive Raji cells. Polyclonal anti-gp72, a C3-binding membrane component, not related to the EBVR but also expressed on the Raji cell surface, was used as a control. Binding of rabbit IgG and EBV on cells was assessed by using immunofluorescence techniques with analysis by flow cytofluorometry. A semiquantitative bioassay was also used to measure the EBV binding. Polyclonal monospecific anti-gp140 IgG inhibits directly the binding of EBV to Raji cells at the same concentration that inhibits the binding of EC3d on cells, whereas a 35 times higher concentration of anti gp72 IgG or preimmune serum IgG does not. Anti-gp140 IgG treatment also inhibits the induction of EBV-determined nuclear antigen in normal tonsil B lymphocytes or in EBV-negative Ramos cells, whereas high concentrations of anti-gp72 IgG or preimmune serum IgG have no effect. These data strongly suggest that gp140, the CR2 of human B lymphocytes, is also the EBVR. PMID- 2983349 TI - Characterization of a protein, released by the T47D cell line, immunologically related to the major envelope protein of mouse mammary tumor virus. AB - The T47D human mammary adenocarcinoma cell line in vitro releases viral particles as well as soluble proteins. Both fractions were shown to contain antigens that immunologically crossreact with the major glycoprotein (gp52) of mouse mammary tumor virus. The crossreacting antigens are located on polypeptides with apparent molecular weights of about 68,000 and 60,000. The larger one is present in viral particles whereas both are found in the soluble fraction. Both proteins are glycosylated. The human tissue culture proteins differ from gp52 not only in molecular weight but also in charge heterogenity and in polypeptide profiles obtained after partial proteolysis. The results suggest that there is a restricted similarity between MMTV gp52 and the immunologically related T47D proteins. PMID- 2983350 TI - Leukotriene B5 is formed in human neutrophils after dietary supplementation with icosapentaenoic acid. AB - Incorporation and conversion of icosapentaenoic acid (20:5, n - 3) by human polymorphonuclear leukocytes were studied in volunteers (n = 6) ingesting a normal Western diet supplemented with icosapentaenoic acid (approximately equal to 4 g daily). Ingestion of icosapentaenoic acid leads to formation of biologically less active leukotriene B5 (LTB5) from polymorphonuclear leukocytes (PMNL) stimulated with ionophore A23187. LTB5 was identified on HPLC by UV absorption and by GC/MS and showed a behavior identical to that of in vitro synthesized LTB5 produced by incubation of human PMNL with icosapentaenoic acid. The ratio of icosapentaenoic acid/arachidonic acid (20:4, n - 6) in cellular phospholipids increased from 0.045 during control to 0.28 after the supplemented period. LTB5 increased from undetectable values to 70.2 +/- 18.7 pmol of LTB5 per 10(7) PMNL during the experimental period. Synthesis of LTB4 did not change significantly (control, 218.8 +/- 89.1; icosapentaenoic acid-enriched diet, 253.6 +/- 18.7 pmol per 10(7) PMNL). The ratio of LTB4/LTB5 corresponded to the ratio of arachidonic acid/icosapentaenoic acid in PMNL phospholipids. Our findings prove that LTB5, which is 10 to 30 times less potent than LTB4 to cause aggregation, chemotaxis, and degranulation of PMNL, can be formed in vivo in man after dietary icosapentaenoic acid. This may modify the contribution of leukotrienes in processes in which these metabolites are of pathogenetic relevance. PMID- 2983352 TI - A comparison of the leukotriene synthesizing ability of subfractions of rat liver glutathione S-transferases. AB - The cytosolic glutathione S-transferases of rat liver have been partially purified by affinity adsorption to glutathionyl Sepharose and elution with glutathione, and have been fractionated by chromatofocusing. The column eluates were pooled into ten fractions, although evaluation of the enzymatic activities of the column eluates with different chromogenic substrates indicated that some of the fractions were heterogeneous. The pooled fractions were characterized with respect to their substrate specificity, their susceptibility to inhibition by several inhibitors, and their ability to catalyze the conjugation of glutathione to leukotriene A4. It was found that all the fractions were able to catalyze leukotriene C formation. The fraction having the highest specific activity with three different chromogenic substrates also had the highest specific activity when LTA was used as the substrate while, in general, there were marked differences in the relative activities of the different pooled fractions. The most active fraction represented approximately 50% of the total glutathione S transferase activity in the whole preparation and had an apparent isoelectric point of 9.05. There was no apparent relationship between the ability of the different fractions to utilize LTA and any of the other substrates which were tested. PMID- 2983351 TI - GABAB-receptor-activated K+ current in voltage-clamped CA3 pyramidal cells in hippocampal cultures. AB - GABAB receptors are a subclass of receptors for gamma-amino-n-butyric acid (GABA) that are also activated by the antispastic drug beta-p-chlorophenyl-GABA (baclofen). One effect of baclofen is to inhibit excitatory transmission from CA3 to CA1 hippocampal pyramidal cells. To identify the ionic mechanism of GABAB receptor-mediated depression, we have studied the effect of baclofen and GABA on ionic currents in voltage-clamped CA3 pyramidal cell somata in rat hippocampal slice cultures. Baclofen (10 microM) induced an inwardly rectifying outward current that reversed at -74 +/- 4.3 mV (mean +/- SD). This appeared to be a K+ current since (i) its reversal potential showed the expected shift when extracellular K+ concentration was changed and (ii) it was blocked by external Ba2+ or internal Cs+. The action of baclofen was closely imitated by GABA after the GABAA-mediated Cl- current had been abolished with pitrazepin (10 microM); under these conditions, GABA (100 microM) also produced an inwardly rectifying, Ba2+-sensitive current with a reversal potential identical to that of the baclofen-induced current. When outward currents were blocked with internal Cs+, the residual inward voltage-dependent Ca2+ current was not changed by baclofen. It is concluded that the primary effect of GABAB-receptor activation in these neurones is to increase K+ permeability rather than to reduce Ca2+ permeability. PMID- 2983353 TI - Effect of PGF-2 alpha on progesterone production in swine luteal cells at different stages of the luteal phase. AB - Suspensions of luteal cells were prepared by enzymatic dispersion of pig corpora lutea obtained at specific times during the estrous cycle. Luteal cells from early corpora lutea produced more progesterone (4.73 +/- 0.84 nmol/10(6) cells, day 3) than those from late diestrus (0.73 +/- 0.04 nmol/10(6) cells, day 15); (P less than 0.05). Bovine LH enhanced progesterone production in a dose dependent manner particularly in cells from 9 to 15 day corpora lutea. Also PGF-2 alpha enhanced progesterone output in cells from mid-late corpora lutea. PGF-2 alpha did not exert any antigonadotropic effect since it further increased the progesterone production induced by LH. Luteal cells produced PGF-2 alpha with levels ranging between 1.6 and 2.7 pmol/10(6) cells throughout the whole luteal phase. The cellular content of cAMP was markedly increased by LH (556 +/- 60%) while it was not affected by PGF-2 alpha. Plasma membrane receptors for PGF-2 alpha were not detected in the analyzed tissue. PMID- 2983355 TI - [Bulk in human nutrition--actions and adverse effects]. PMID- 2983354 TI - On the synthesis of prostaglandins E1, E2 and E2 alpha by sow oviducts. Differential modulation of 1 and 2 series of prostaglandins by norepinephrine. AB - The present study was performed in order to evaluate whether norepinephrine (NE) can modulate the synthesis and release of 1 and 2 series of prostaglandins (PGs) by the isthmic region of preovulatory sow oviducts and also to clarify whether the action of the neurotransmitter is mediated through alpha, through beta or through both types of tissue adrenoreceptors. NE, at a concentration of 1 microgram/ml, depressed significantly (P less than 0.05) the basal output of PGE1 and enhanced (P less than 0.01) the release of PGE2 but, did not modify, the output of "PGF2 alpha-like material". Propranolol (10(-7)M) failed to alter the basal output of "PGE1, PGE2 or PGF2 alpha-like material". In the presence of this beta-adrenoreceptor blocker, the depression induced by NE on PGE1 output, was abolished; its stimulatory influence on the release of PGE2, was eliminated and no effect was detected regarding PGF2 alpha. On the other hand, phentolamine (10( 6)M) did not alter the basal output of "PGE1, PGE2 or PGF2 alpha-like material" and also failed to modify the depression induced by NE on PGE1 release. However, this alpha adrenoceptor blocker completely inhibited the stimulatory action of NE on the output of PGE2 into the incubating medium. The foregoing results document opposite actions of NE on PGE1 and PGE2 outputs from the isthmic region of proestrous sow oviducts and suggest the involvement of beta-adrenoreceptors in both disparate influences. The activation of alpha adrenoreceptors also appears associated with the enhancing effect of the agonist on the release of PGE2. The possible physiological significance of these findings is discussed in terms of the function of the isthmic region as an "adrenergic sphincter" able to influence ovum transport around the moment of ovulation. PMID- 2983356 TI - Involvement of a GABAergic mechanism in the anticonvulsant effect of pentobarbital against maximal electroshock-induced seizures in rats. AB - The interaction between pentobarbital and other modulators of GABAergic transmission (diazepam, ethanol and progabide) was investigated on maximal electroshock seizures and on the loss of righting reflexes in rats. Pentobarbital, diazepam and ethanol produced a dose-dependent protection against electroshock seizures, with pentobarbital being more potent (3- and 50-times) than diazepam and ethanol. Progabide neither provided protection nor caused loss of righting reflex. Subprotective doses of pentobarbital and diazepam, together or when combined with a single ineffective dose of ethanol or progabide, caused protection against seizures and loss of righting reflex for variable durations, while ethanol and progabide combination did not provide protection. The protective effect of diazepam was antagonized by RO15-1788, picrotoxin and bicuculline pretreatments. The antagonism of pentobarbital protection by a specific GABA receptor antagonist, bicuculline suggests involvement of the GABAergic system in the anticonvulsant effect of pentobarbital. These results indicate that, like diazepam, the anticonvulsant effect of pentobarbital appears to be mediated through a GABAergic mechanism. PMID- 2983357 TI - Effect of long-term administration of delta 9-tetrahydrocannabinol on hepatic mixed-function oxidase systems in the rat. AB - Chronic oral treatment of young female Fischer rats with 25 mg/kg delta 9 tetrahydrocannabinol (THC) per day inhibited aminopyrine demethylation and significantly increased benzo[a]pyrene oxidation by the liver. THC treatment also elevated serum corticosterone levels and produced a significant loss of body weight. The weight loss was not due to vehicle or food intake (pair-feeding). Pair-feeding did, however, produce a stimulation of both mixed function oxidase pathways as well as a marked elevation in serum corticosterone levels. The results indicate that THC has a differential effect on mixed function oxidase pathways in the liver that is not directly related to food intake or corticosterone levels. PMID- 2983359 TI - Study of adenosine receptors in intact rat fat cells by radioligand binding. AB - Binding of (-)N6-phenylisopropyl[3H]adenosine ([3H]PIA) to intact rat fat cells was studied in the presence of the adenosine uptake blocker dipyridamole. Specific binding of 5 nmol/l [3H]PIA at 37 degrees C was rapid, reversible and dependent on cell concentration and the presence of adenosine deaminase. Saturability of specific binding was not achieved at concentrations up to 200 mumol/l [3H]PIA. In competition experiments (-)PIA (IC50 42 nmol/l) was the most potent analogue, followed by 2-chloroadenosine and 5'-N ethylcarboxamidoadenosine. Binding of [3H]PIA was stereospecific, since (-)PIA was 200 times more potent than (+)PIA. The adenosine antagonist theophylline inhibited binding with an IC50 of 16.9 mumol/l, whereas adenine, inosine and GTP did not affect binding. The results allow us to describe several characteristics of [3H]PIA binding to intact fat cells but a considerable component of nonreceptor binding impedes a detailed study of adenosine receptors under physiological conditions. PMID- 2983358 TI - Cardiovascular effects of dobutamine and converting enzyme inhibition in rats with diabetic ketoacidosis. AB - We investigated cardiac and peripheral hemodynamics in rats with streptozotocin induced diabetes mellitus in comparison to normal controls. Diabetic rats showed a significantly decreased cardiac output and heart rate, with unchanged mean blood pressures. Regional blood flows were unchanged except for renal blood flow which was significantly lower. The low output state associated with diabetes was reversed by angiotensin converting enzyme inhibition with MK-422, but not with dobutamine, although both agents caused similar reduction in peripheral vascular resistance. However, despite improved left ventricular performance, renal perfusion did not improve after MK-422. PMID- 2983361 TI - Transplant of an inferiorly based rectus abdominis myocutaneous flap to the calf. PMID- 2983360 TI - Histoautoradiographic localization of (-)-3H-dihydroalprenolol in rabbit aorta. AB - The distribution pattern of (-)-3H-dihydroalprenolol (3H-DHA) binding sites within the thoracic and abdominal aorta was studied using a histoautoradiographic technique. Frozen sections of rabbit aorta were incubated with 3H-DHA in the presence or absence of 1 mumol/l (-)-propranolol and of increasing concentrations of various adrenergic agents. After washing, the same secretions were placed in scintillation vials and counted to evaluate the pharmacological characteristics of 3H-DHA binding or processed for histoautoradiography. 3H-DHA was bound by sections of the rabbit aorta in a manner consistent with the presence of beta adrenoceptors. The binding was reversible, saturable and of high affinity, with a dissociation constant Kd of 1.0 nmol/l and a maximum binding capacity Bmax of 17 fmol/mg tissue (wet weight). It was inhibited only by beta-adrenoceptor blocking drugs. Histoautoradiographic studies revealed that beta-adrenoceptors are located primarily in the media and then in the intima. The lowest density of 3H-DHA was found in the adventitia. The distribution pattern of beta-adrenoceptors in the media suggests the possible existence of two groups of beta-adrenoceptors within this layer of the aorta. The first group was found in the outer portions of the media and may be controlled by the adrenergic nerves that supply the artery. The second group was found in the portions of the media closest to the intima and is likely under the control of catecholamines coming from the lumen of the aorta. PMID- 2983362 TI - Activation of endogenous C-type retroviral genomes by internal alpha-irradiation of mice with 224Radium. AB - Sensitive cocultivation techniques were applied to study the radiation-induced activation of endogenous retroviral genomes in different mouse strains by the alpha-emitting radionuclide 224Radium. Activated infectious C-type retroviruses were detected in spleen, bone marrow and bone tissues of C57BL/6-, BALB/c- and NMRI mice. The titres of high-dose-irradiated animals were higher than those found in low-dose-irradiated animals. Infectious retrovirus could be detected with a dose of 13.2 rad (maximum dose rate 0.9 rad/day) in the skeleton, and a dose of 4.2 rad (maximum dose rate 0.3 rad/day) in the spleen. The virus activation pattern was different in the three mouse strains. These data indicate that activation of endogenous retroviral genomes by alpha-irradiation shows a dose-effect relationship and a dependence on the genetic background of the mouse. PMID- 2983363 TI - A sensitive SV40 viral probe assay for DNA strand breaks and their biological repair in higher cells: techniques and preliminary results. AB - An alternative experimental approach to the investigation of intracellular repair of DNA strand breaks by mammalian higher cells has been developed using simian virus 40 (SV40), which has no known intrinsic DNA repair capacity and possesses a minichromosome structure, as an intracellular probe. In this approach unirradiated simian or human cells are infected with irradiated virus and incubated for varying periods. Nuclei are isolated, and viral DNA is extracted and assayed for residual damage. This assay involves separation of the viral DNA by agarose gel electrophoresis into three sharply demarcated bands corresponding to DNA molecules containing a double-strand break (DSB), single-strand breaks (SSB), or no breaks. Quantitative data are obtained by a combination of DNA hybridization with 32P-labeled SV40 DNA, autoradiography, and densitometry. Various experiments have been carried out to investigate the feasibility of this approach and to establish the optimal experimental conditions for its use. These experiments indicate that there is rapid and efficient cellular uptake of SV40. independent of prior radiation dose to the virus, and that this multistep experimental procedure gives excellent recovery and quantitation of the three DNA forms when compared with more direct methods of measurement. Radiation dose response experiments with purified extracellular SV40 virus, using this approach, are quite reproducible and give results closely comparable to those obtained with techniques in current use. Initial time-course incubation experiments with SV40 infected CV-1 monkey kidney cells indicate that this approach can demonstrate slow but extensive intracellular repair of SSB; and limited presumptive early repair of DSB, followed by later and more extensive induction of DSB. PMID- 2983364 TI - [Role of the low-molecular components of the blood serum in determining the biological behavior of plutonium]. AB - A study was made of the ability of plutonium to bind to a complex with amino acids, organic and inorganic acids of normal blood serum. Among the low molecular weight addends bicarbonate ions play a major role in plutonium binding. In this respect, possible mechanisms of plutonium metabolism in the blood are discussed. PMID- 2983365 TI - [Acute toxicity and radioprotective effectiveness of gammaphos on intramuscular administration to mice]. AB - In experiments on mice it was shown that acute toxicity of gammaphos (WR-2721) was 790 mg/kg and 862 mg-kg after intraperitoneal and intramuscular administration, respectively. Gammaphos in the dose of 100 mg/kg, injected intramuscularly, increased the radioresistance of mice in nearly the same way as cystamine, in the dose of 150 mg/kg, did. The increase in the dose of gammaphos up to 200 or 300 mg/kg, injected intramuscularly, enhanced the radioprotective effect. No change was observed in the radioprotective efficiency of gammaphos after intramuscular injection as compared to intraperitoneal administration of the protective agent in the same dose. PMID- 2983367 TI - Abnormalities of the neonatal cerebral ventricles. AB - High-resolution ultrasonography allows accurate assessment of ventricular size and contour in the infant, so that conditions such as hydrocephalus, holoprosencephaly, and agenesis of the corpus callosum can be diagnosed. Ventricular shunt catheter placement can be guided, and some of the complications of cerebrospinal fluid shunt systems can be identified. Infection of the ventricular fluid causes striking findings, including increased echogenicity and septation within the lumen. PMID- 2983366 TI - [Postradiation inhibition of poly ADP ribose polymerase is not determined by internucleosomal fragmentation of DNA]. AB - Postirradiation changes in poly(ADP-ribose) polymerase activity in nuclei and permeable thymocytes were studied. The incorporation of 14C-NAD into poly(ADP ribose) was virtually invariable during the first 60 min after irradiation at a dose of 10 Gy, but after 2 and 3 h it made up, as compared to the control, 60 and 45% for nuclei, and 65 and 55% for permeable thymocytes, respectively. It was shown that the internucleosomal fragmentation of DNA was not responsible for the observed changes in poly(ADP-ribose) polymerase activity. PMID- 2983368 TI - Applications of ultrasound in the neonatal abdomen. AB - In this article, the uses of real-time sonography relative to evaluation of the neonatal abdomen are presented. Included are some of the more recent applications that, to a great extent, have evolved because of the ease with which portable real-time examination of the fragile or compromised infant can be accomplished. PMID- 2983369 TI - Whole-breast sonography. AB - Ultrasound has been found to be useful as an adjunct to mammography and physical examination of the breast. It has detected lesions not identified by any other modality and has allowed more precise diagnosis of palpable and/or radiographically demonstrated lesions. Ultrasound should not be used as the sole breast imaging modality, however, because of its inability to detect microcalcifications and its difficulty in demonstrating small solid lesions, particularly in the fatty breast. PMID- 2983370 TI - Ultrasonography of the salivary glands. AB - Sonography and computed tomography play complementary roles in imaging mass lesions in the salivary glands and their surroundings. Ninety per cent of parotid tumors originate from the superficial lobe, and we consider sonography to be the method of choice for imaging these lesions. The sensitivity of high-resolution sonography in detecting intraparotid tumors approaches 100 per cent, and ultrasound is therefore an excellent method to evaluate patients with parotid swelling. It may provide clinically useful information by precisely outlining the tumor borders or by detecting multiple or bilateral lesions. We restrict our use of CT to tumors that appear to extend beyond the borders of the parotid gland with possible invasion of surrounding soft tissues or bone. CT is also the method of choice for differentiating lesions of the deep lobe from parapharyngeal tumors and for staging carcinomas. Although ultrasound supplemented with computed tomography should replace conventional sialography in the work-up of parotid neoplasms, sialography remains the method of choice for evaluating patients with chronic sialadenitis, autoimmune diseases, and sialolithiasis. The role of sonography in these conditions is limited to ruling out a parotid neoplasm, assessing the extent of abscess formation, or assisting in localizing calculi in selected patients. PMID- 2983371 TI - Neurosurgical applications of intraoperative ultrasound. AB - Intraoperative sonography is becoming increasingly important in neurosurgery. In this article, the authors provide step-by-step instructions, the basics of lesion localization, sonographic characteristics of the more common brain and spine lesions, and tables of references to sonographically described lesions in the literature. PMID- 2983372 TI - Lung cancer and pregnancy. AB - Two pregnant patients presenting with bronchogenic carcinoma are described. A retrospective study of patients with bronchogenic carcinoma as well as a review of the literature attest to the rarity of this combination. If current smoking patterns continue, the association of pregnancy and bronchogenic carcinoma will increase in frequency. PMID- 2983373 TI - Parathyroid adenomas evaluated by Tl-201/Tc-99m pertechnetate subtraction scintigraphy and high-resolution ultrasonography. AB - Thallium-201/technetium-99m pertechnetate subtraction scintigraphy of the parathyroid glands was performed in a prospective study of 33 patients who had undergone bilateral neck exploration for elevated serum calcium and serum parathyroid hormone levels. In 31 cases, the Tl-201/Tc-99m subtraction technique yielded an overall sensitivity of 81%, specificity of 99%, and accuracy of 94% for identifying solitary parathyroid adenomas. Tl-201/Tc-99m subtraction scintigraphy correctly identified 73% of parathyroid adenomas weighing less than 499 mg, 79% of those weighing 500-1,499 mg, and 100% of adenomas weighing more than 1,500 mg. In a subgroup of 24 patients with solitary parathyroid adenomas who underwent both scintigraphy and high-resolution sonography, the sensitivity, specificity, and accuracy of both procedures were similar. PMID- 2983374 TI - Popliteal cysts and deep venous thrombosis: Tc-99m red blood cell venography. AB - Thirty-four patients who clinically were suspected of having deep venous thrombosis (DVT) underwent Tc-99m red blood cell (RBC) venography followed by contrast venography. The sensitivity (88%) and specificity (94%) of Tc-99m RBC venography for DVT confirmed findings of previous studies. Twenty-four patients who had proved popliteal cysts and swollen calves also were examined using Tc-99m RBC venography. Large cysts (greater than 9 X 4 X 4 cm) showed lack of DVT features, photon deficient cold areas in the popliteal fossa, and diversion of venous flow around the photon deficient area. These features were seen in 25% of the study population (six patients), and assisted in the distinction between the two conditions. PMID- 2983375 TI - Hepatoma, arterioportal shunting, and hyperkinetic portal hypertension: therapeutic embolization. AB - Four patients with hepatocellular carcinoma, shunting of blood from the hepatic artery to the portal vein, and hyperkinetic portal hypertension were treated by transcatheter embolization of the hepatic artery. In three acutely bleeding patients variceal hemorrhage was controlled by the embolization. Following embolization hepatofugal portal venous flow became hepatopetal in all four patients. No serious complications were encountered. When hepatoma is complicated by arterioportal shunting and hyperkinetic portal hypertension, occlusion of the fistula by transcatheter embolotherapy can reduce the portal pressure. PMID- 2983376 TI - The influence of prostaglandins on neurotransmission in the rabbit isolated vas deferens. AB - Arachidonic acid and PGs of the D, E, F and I series were examined for influences on neurogenic contractions of the rabbit isolated vas deferens. This preparation exhibits two pharmacologically distinct contractions in response to electrical stimulation. All of the PGs tested inhibited the neurogenic contractions but the pattern of inhibition differed. PGE1 and PGI2 inhibited the adrenergic contractile phase more potently than the nonadrenergic, and PGF2 alpha exhibited the opposite selectivity. Arachidonic acid, PGE2 and PGD2 produced equipotent effects on both contractile phases, although PGE2 was the most potent in producing these effects. None of the PGs altered the concentration-response curve to norepinephrine. Contractile responses to ATP, a putative neurotransmitter, were inhibited by PGF2 alpha but not by the other PGs. These results suggest that the PG effects are predominantly prejunctional. The differing potencies of the PGs on the two neural components are consistent with the hypothesis that neurotransmitters in the vas deferens are released by distinct types of nerves. PMID- 2983377 TI - Synthesis of prostaglandins and leukotrienes. Effects of ethanol. AB - Prostaglandins, thromboxane, and leukotrienes are metabolites of arachidonic acid that have a variety of physiological effects at low concentrations. Effects include smooth muscle contraction, platelet aggregation, platelet disaggregation, bronchoconstriction, increased capillary permeability, cardiac dysfunction, and polymorphonuclear leukocyte accumulation. Although their formation does not appear to be essential for life, these eicosanoid activities are wide ranging and have important roles in normal physiology as well as pathophysiology. At the center of eicosanoid biosynthesis lies the plasma cell membrane which serves as the arachidonic acid reservoir. It has been widely appreciated that ethanol exerts effects on the lipid bilayer, and it is not surprising that a growing body of evidence supports the concept that important interactions between ethanol and eicosanoid biosynthesis can occur. Furthermore, at various steps leading to ultimate prostaglandin, thromboxane and leukotriene production, reactive intermediates such as radicals are involved whose lifetime in the biological milieu can be profoundly modulated by ethanol. PMID- 2983378 TI - Biochemical interactions of ethanol with the arachidonic acid cascade. AB - A rapidly increasing scientific literature now supports the possibility of an alcohol-prostaglandin interaction. This chapter reviews evidence for both direct and indirect biochemical interactions between ethanol and the metabolism of arachidonic acid and several related compounds. Much of the present data is based on pharmacological manipulation of prostaglandin (PG) levels by potent nonsteroid anti-inflammatory agents such as indomethacin. Indomethacin markedly alters the behavioral response to ethanol, particularly in the mouse model. These data suggest that PGs are involved in the behavioral response to acute ethanol exposure in the mouse. In other animal models, alcohol has been reported to alter blood platelet metabolism of arachidonic acid, to suppress the enzymatic degradation of PGs, and to alter the response of the adenyl cyclase system to several hormones including PGs of the "E" series. In humans, both the stimulation and inhibition of PG synthesis is reported to aid the treatment of various aspects of alcoholism. Further, PGs are reported to protect against alcohol induced fatty liver, and both PGs and arachidonic acid protect the gastric mucosa against ethanol-induced lesions. Certainly the residual consequences of acute, excessive ethanol consumption are commonly treated with a prostaglandin synthesis inhibitor. The material in this chapter is an attempt to review the data and to discuss the molecular mechanism underlying these observations. PMID- 2983379 TI - Functional brain imaging. AB - Recent advances in brain imaging have allowed a regional examination of brain function using multiple-probe inert gas studies of cerebral blood flow, positron or single photon tomography. Inert gas blood flow methods using inhalation or injection of 133xenon have been used with multiple-probe systems to measure blood flow in 1 to 2 cm regions of lateral cortex. The sensitivity of these systems to neurophysiological stimuli and neurological diseases have been demonstrated in numerous studies of the normal resting state, memory and learning, motor activity and sensory input, dementia, and aphasia, to name some. Positron tomography utilizes cyclotron-produced, short-lived positron-emitting isotopes to label biologically active radiopharmaceuticals. Using positron tomographs capable of quantitative three-dimensional imaging and appropriate tracer-kinetic models, regional metabolic function, including glucose, oxygen, amino acid metabolism, and receptor-binding can be regionally studied throughout the brain. Clinical studies have been performed in dementia, schizophrenia, affective disorders, resting states, and sensory stimulation. Positron tomography offers potentially the greatest variety of studies and highest temporal and spatial resolution of any of the presently available functional brain-imaging modalities. Its principal drawback is the very high cost. Single photon tomography uses gamma-emitting isotopes such as 123iodine and 133xenon to image regional cerebral blood flow and recently receptor function. Although at present it does not have the variety of studies or the technical capabilities of positron tomography, it does provide three-dimensional studies with 1 to 2 cm resolutions throughout the brain at a considerably lower cost than positron tomography. In the future, magnetic resonance studies of blood flow or phosphorus metabolism may add a fourth modality. PMID- 2983380 TI - [Effect of the contrast medium on the gray shading in magnetic resonance tomography]. AB - Contrast media can significantly improve the diagnostic value of MR tomography. The basis of using contrast material for MR depends on totally different physical principles from those employed in conventional radiological diagnosis. The physical factors underlying the use of these contrast media are described, various suitable substances are discussed, and experience so far gained with some contrast agents is outlined. PMID- 2983381 TI - [Diagnosis of bone necrosis by magnetic resonance tomography. The potentials for early diagnosis]. AB - Twenty-eight patients with necrosis in the hip joint, the femoral condyles and the talus were examined by MR tomography. This revealed various features whose frequency depended upon the various stages of the necrosis. Three patients who showed no, or only indefinite, changes on radiographs, displayed clear evidence of necrosis on MR. Staging of femoral head necrosis on the basis of MR criteria is suggested, as well as a suitable procedure for early diagnosis. Accurate localisation and knowledge of the extent of the necrosis by means of MR makes it possible to plan an osteotomy accurately. PMID- 2983382 TI - [CT and ERCP for the combined study of biliary tract diseases]. AB - The value of combined CT and ERCP for examining the bile ducts was studied retrospectively. CT on its own led to a correct diagnosis in 59% of cases. CT was valuable for estimating the calibre of the bile ducts, for localising the level of obstruction, for demonstrating calcified biliary concrements, and for showing peribiliary tumours. ERCP yielded a correct diagnosis in 67% of cases. The value of ERCP lies in the demonstration of poorly calcified and small concrements, strictures, tumours with ductal infiltration, and for investigating the post cholecystectomy syndrome. Combining these two types of examination increases diagnostic accuracy to 82%. PMID- 2983383 TI - [Diagnostic potential and value of conventional radiography, x-ray tomography and high-resolution computed tomography in complex orbital trauma]. AB - Hypocycloidal polytomography and High Resolution CT (HR-CT) were used in 101 severely injured patients having complex orbital and periorbital trauma. The diagnostic value of both methods was analysed retrospectively. Biplane HR-CT was superior to biplane polytomography especially in diagnosing complex fractures and orbital soft tissue lesions. Scanned coronal or semicoronal projection was performed in 70 out of 101 cases and found mandatory for evaluation and diagnosing of undisplaced floor fractures. Recognition of traumatic orbital soft tissue lesions was possible by CT and found extremely helpful in diagnosing secondary and tertiary fractures, primarily not depicted on polytomograms, but detected retrospectively in some cases. Many of these lesions correlated well with various posttraumatic ophthalmological impairments. PMID- 2983384 TI - [Computed tomographic and clinical follow-up studies in intraventricular hemorrhage]. AB - The investigation comprises 41 children with intraventricular hemorrhage (IVH). The CT-findings were divided into 4 groups, according to Papile. A CT and clinical follow-up study was performed on the survivors. The grade of IVH correlated with the rate of mortality and with neurological deficits. It is demonstrated in 3 cases that the CT-scan is only part of a prognostic aspect which is remarkable influenced by clinical events. PMID- 2983385 TI - [Measurement of regional cerebral blood flow as a supplement to carotid angiography in assessing cerebrovascular insufficiency]. AB - The examination of 91 patients was undertaken in order to establish the relevance of tomographical measurement of the regional cerebral blood-flow, following inhalation of 133Xenon, as a complementary method to the conventional carotis angiography. In many cases the findings revealed a more precise or even deviating judgement on the hemodynamic relevance of stenosis and obliteration than was proved only by angiography. Especially in the stages I and IV of cerebrovascular disease of the a. cerebri anterior, a. carotis interna and a. carotis communis the rCBF-measurement seemed to be an important parameter for the decision between conservative and surgical therapy as a prognostic parameter for the individual risk of many patients to suffer a cerebral infarction. PMID- 2983386 TI - [Reduction of the rate of cerebral angiography complications by a standardized study method]. AB - The complications from cerebral angiography encountered in 1034 patients between 1977 and 1982 are described. A standardised technique of cerebral transfemoral catheter angiography is outlined, which helps to reduce the number of complications. The role of the disease for which the examination is done, and the effect of the new contrast media on the risk of the examination, are discussed. Patients with a compromised blood-brain barrier and manifest reduction in cerebral blood flow run a greater risk than patients with other abnormalities. Low osmolar contrast media are better tolerated subjectively. Theoretically, their use would be expected to lead to a reduction in complications but, so far, it has not been possible to confirm this. PMID- 2983387 TI - [Computer tomography and spinal angiography in the diagnosis of spinal angioma]. AB - The computer tomographic and angiographic features of spinal angiomas are described. The authors observed 21 cases and, on the basis of this material, they deal with the clinical features, radiological appearances, disease patterns, forms of treatment, and prognosis. CT and angiography produce a fairly uniform and typical appearance, permitting diagnosis before therapy. Problems in differential diagnosis and errors in interpretation are enumerated. PMID- 2983388 TI - [Changes in the cervical spine in chronic polyarthritis]. AB - In 93 patients with classical chronic polyarthritis (rheumatoid arthritis) (at least five ARA-criterias) there were inflammatory lesions of dens epistropheus in 48.4%, ventral atlantoaxial subluxations in 25.8%, lateral atlantoaxial dislocations in 14% and pseudobasilary invagination in 5.4%. Step-ladder subluxation between C 2/C 7 was found in 31.2%, discitis in 12.5% and spondylarthritis in 38% of cases. Inflammatory signs of the cervical spine were correlated to the Steinbrocker-Grade IV, ANA level 1:40 and the degree of Waaler Rose. The correlation between long standing steroid-therapy and signs of cervical involvement during c.p.--specially in C1/C2--is found to be proven. There are connections between the duration of c.p. (more than 10 years) and manifestation of cervical spine lesions, further, in a protective sense between longstanding gold therapy and cervical spine lesions. PMID- 2983389 TI - Rheumatoid arthritis of the shoulder. Description and standard radiographs. AB - The course of rheumatoid arthritis in the shoulder is evaluated in 143 patients. In a period of 29 years, 630 x-rays were taken of 286 shoulders. In this series 2 or more x-rays per shoulder were taken of 89 patients (29 male, 60 female). The various changes in the glenohumeral and acromioclavicular joints are described. Gross destruction appears to be rare, compared to the more frequently seen minor cystic changes. The progress of the disease is often slow or halting. One or both of the shoulders in some of the patients (15 male and 29 female) did not have any detectable x-ray changes, although some of them were followed up for more than 20 years. During our follow-up it became apparent that the acromioclavicular and glenohumeral joints do not follow the same course neither in time nor in severity of joint destruction. Therefore, we divided the shoulder joint into the acromioclavicular and glenohumeral joint. One normal stage and 5 stages of pathology are recognised to fit into previously published schemes of the other joints. Stage 5 appears to be a new phenomenon of neojoint formation, under the previous humeral head with the inferior glenoid rim. Joint disease in the acromioclavicular joint could be divided only into 3 stages. PMID- 2983390 TI - [Reparative giant-cell granuloma of the extremities]. AB - Giant-cell reparative granulomas are observed not infrequently in the mandible as sharply demarcated osteolytic lesions. On the other hand, these tumour-like lesions occurring in short tubular bones are rarely reported in the literature and, so far, only 27 cases have been described. We have records of five cases in the Westphalian Bone Tumour Register. The actual occurrence is probably greater if one assumes that the lesion is caused by an intra-osseous bleeding. This pathogenic mechanism is supported by histological evidence. PMID- 2983391 TI - [Bone changes in Hansen's disease]. AB - Bone lesion is a frequent organic manifestation in leprosy. Osseal destructions caused by granulomatous process induced by M. leprae are so-called specific lesions in contrast to non-specific lesions based on nerval or arterial diseases. The specific osseal alterations are characterized by cystic brightenings in roentgenograms while non-specific osseal changes show absorption to bone structure as akroosterolysis and osteoporosis. Typical radiologic findings in different stages of mutilation are demonstrated. PMID- 2983392 TI - Antegrade pyelography, percutaneous nephrostomy and ureteral perfusion (Whitaker test) for the renal transplant recipient. AB - Ureteral obstruction is a serious complication in renal transplant recipients. The diagnosis may be difficult with standard methods of investigation, and definite treatment may, therefore, be delayed. This paper describes the indications, the technique, and the experience with antegrade pyelography, ureteral perfusion and percutaneous drainage in 18 patients with suspected urinary obstruction. Though invasive, these methods proved to be a safe, fast and accurate means for evaluating the location and significance of ureteral obstruction. For initial treatment in obstruction, percutaneous nephrostomy is the method of choice. Surgery should be delayed until recovery of renal function with decreasing serum creatinine is demonstrated. Patients with persistent or rising serum creatinine (greater than 250 mumol./l.) did not benefit from surgical relief of obstruction. PMID- 2983393 TI - [Local thrombolysis of renal artery embolism]. AB - Thrombo-embolic occlusions of the renal arteries are usually a result of co existing cardiac disease, in most cases cardiac arrhythmias or ischaemic heart disease. Similar findings have been observed in mesenteric infarction and embolism into the lower extremities. The duration of ischaemia determines the prognosis. Surgical intervention has been the mainstay of previous therapy. However, the application of local thrombolysis, which is an extension of the diagnostic catheter studies, provides an alternative form of treatment. As in the case of peripheral artery disease, the choice of treatment can only be made in consultation with the surgeon. This report presents experience with the use of local thrombolysis of renal artery emboli. PMID- 2983395 TI - [Animal experiment results with a new vena cava filter]. AB - A new cava filter has been tried in 23 dogs weighing between 16 and 23 kg. The filter consists of a stainless steel basket which can be introduced either antegrade or retrograde through a 10F teflon catheter and which can be removed again within the first four days. The filter was effective in holding back embolising thrombus. In the early stages, even without artificial embolisation, two out of five animals showed clot formation in the basket. After three or four months, all the filters were permeable and on only two occasions was a small amount of thrombus found. PMID- 2983394 TI - [Results and experience with urethrocystography in diagnosing female stress incontinence]. AB - The normal parameters of urethrocystography were evaluated in 324 examinations. There were definite differences between continent and pressure-incontinent women on the one hand, and stress-incontinent patients on the other, as regards the pubo-urethral angle, but not as regards the posterior vesico-urethral angle. Preoperative and post-operative urethro-cystograms showed statistically significant differences. PMID- 2983396 TI - [Effect of the method of measurement on the interpretation of renal clearance]. AB - The concept of renal clearance measured during steady-state conditions includes the effect of filtration and secretion as well as urine flow. On the other hand, renal clearance, as used in nuclear medicine, only describes the effect of filtration and secretion. A way for determining steady-state renal clearance with a bolus injection technique is described. Steady-state renal clearance reflects the efficiency of filtration and secretion, urine flow, and the steady-state chemical potentials in the kidneys. PMID- 2983397 TI - [Simultaneous tomography using higher-sensitivity screens]. AB - Experiments were carried out for performing multiplane tomography using rare earth screens. These screens provided adequately regular exposures at various levels and with good details. Dose reduction of at least 50% can be achieved. PMID- 2983398 TI - [1st report on indirect lymphography using iotasul in the head and neck region]. AB - The early results in 29 patients with tumours in the head and neck are described, on whom indirect lymphography with the water-soluble contrast medium Iotasul had been carried out (experimental contrast medium, Schering AG). After careful sub epidermal injection of about 12 ml contrast, either lateral to the eye or in the region of the chin, lymph vessels were seen in 25 patients and lymph nodes in 20. These were demonstrated by conventional radiography or by xero-radiography. There were no local or generalised reactions. PMID- 2983399 TI - [Secondary aneurysm of the iliac vein as a complication of traumatic arteriovenous fistulae of the lower extremity]. PMID- 2983400 TI - [Anterograde dilatation of a stenotic uretero-enteric anastomosis under endoscopic control]. PMID- 2983401 TI - [Cruveilhier-Baumgarten syndrome without esophageal varices]. PMID- 2983402 TI - Cavernous haemangioma of the spleen. PMID- 2983403 TI - [Adhesive ileus in cecum mobile]. PMID- 2983404 TI - [Unusual occurrence of pneumoperitoneum]. PMID- 2983405 TI - Dietary fiber and cholesterol and bile acid metabolism in axenic (germfree) and holoxenic (conventional) rats. III. Effect of non-sterilized pectin. AB - Axenic (germfree) and holoxenic (conventional) rats were given a pectin containing diet (5%) or a pectin-free diet. The diet was not sterilized and the axenic state was maintained by the addition of antibiotics. Pectin modified little or not at all the quantity of bile acids in the small intestine, fecal bile acid excretion and fecal elimination of the sum of cholesterol and bile acids. It similarly decreased the concentration of liver cholesterol and the amount of bile acids in the cecum and large intestine of both germfree and conventional rats. Pectin decreased the absorption coefficient of dietary cholesterol and its plasma concentration and increased fecal cholesterol elimination in conventional rats but not in germfree animals. The data imply that some of the actions of pectin depend on the presence of the intestinal flora but the exact nature of this relationship is unclear. PMID- 2983406 TI - Humoral antibody response and assessment of protection following primary vaccination of chicks with maternally derived antibody against avian infectious bronchitis virus. AB - Commercially bred chicks with maternally derived antibody to avian infectious bronchitis virus (IBV) were hatched in isolated conditions and a number vaccinated within the first three weeks of life with live IBV strain H120. Humoral antibody responses were assayed by haemagglutination inhibition (HI) or neutralisation (SN) tests, and the degree of protection against challenge with the virulent Massachusetts M41 strain assessed on the basis of tracheal ciliary activity four days after challenge. Maternal antibody in unvaccinated chicks declined linearly with a mean half-life of five to six days based on both HI and SN tests; these chicks were protected against challenge until four weeks old. There was complete correlation between ciliary activity and histopathological findings, but little between protection and antibody titre. It was concluded that the optimum age for primary vaccination was about two weeks. PMID- 2983407 TI - Immunological response of chickens to eastern equine encephalomyelitis virus. AB - The dominant immunoglobulin against eastern equine encephalomyelitis (EEE) virus and its duration and the longevity of the EEE virus haemagglutination inhibition (HI) antibodies were determined in sentinel and 125 immunised and hyperimmunised domestic chickens by enzyme-linked immunosorbent assay and the HI test respectively. The chickens ranged in age from 10 weeks to 18 months, were of varied pedigrees and from different countries. Results show that the HI antibody (IgG) is short-lived. It peaks and disappears within 30 days. The secondary response is dominated by the IgM immunoglobulin which is relatively long-lasting. These results are contrary to classical expectations and were observed in all the chickens studied. If these observations are found to be characteristic of birds generally, the present standard method of EEE virus seroepidemiological surveillance must be modified to be effective. PMID- 2983408 TI - Influence of a 12.5 per cent rapeseed diet and an avian reovirus on the production of leg abnormalities in male broiler chickens. AB - The incidence of different forms of leg abnormality were recorded in reovirus (S1133) infected and control male broiler chickens fed on a normal commercial diet or one of similar nutritive value containing 12.5 per cent rapeseed meal. Regular serological examination showed that birds remained free from Mycoplasma gallisepticum and M synoviae infection throughout the 10 week period of investigation. Precipitating antibodies to the reovirus were detected in 90 per cent of the infected birds between the third and 10th week after infection. Carotene levels in rapeseed fed groups showed no significant differences between reovirus infected and control birds or between birds with or without clinical signs of leg abnormality. The most frequent and severe leg abnormalities were present in the infected birds fed on the rapeseed diet, followed by those fed on the normal commercial diet. There was a highly significant difference (P less than 0.01) between the number of birds with leg abnormalities in each of these groups and their corresponding control groups. The lesions which were mainly responsible for these differences were tenosynovitis and enlarged hocks. Oral infection with reovirus did not appear to make the birds more susceptible to other types of leg abnormality, although the severest lesions of dyschondroplasia were seen in birds which had been exposed to the dual effects of reovirus and rapeseed diet. PMID- 2983409 TI - [Undifferentiated small cell bronchial cancer: role of the medical oncologist]. PMID- 2983410 TI - [Therapy of constipation in geriatric patients]. PMID- 2983411 TI - Are sodium bicarbonate and potassium bicarbonate fully dissociated under physiological conditions? AB - In solutions containing 160 mmol/l Na+ and K+, respectively, measurements with an ion-selective electrode system (KNA1, Radiometer), showed apparent falls in the respective Na+ and K+ concentrations when C1- was replaced by HCO3-. After correction for the change in liquid junction potential, the fall was 9.2 mmol/l for Na+ and 7.3 mmol/l for K+. On the basis of these findings we conclude that sodium bicarbonate and potassium bicarbonate are not fully dissociated in solution, and that NaHCO3(0) and KHCO3(0) do exist as chemical components with association constants of 0.72 and 0.55, respectively. Using these association constants, normal plasma will contain 1.2 mmol/l NaHCO3(0) and 0.03 mmol/l KHCO3(0). Thus NaHCO3(0) accounts for virtually the same amount of CO2 as the physically dissolved fraction. A review of all the currently known CO2 species in plasma suggests that there may be a residue of about 2 mmol/l of unknown CO2 species in normal plasma. PMID- 2983412 TI - Apparently high plasma angiotensin II levels in patients with essential hypertension treated by converting enzyme inhibition. AB - Plasma angiotensin I and II (AI and AII) were measured in 13 patients with essential hypertension before and during chronic treatment with enalapril (MK 421) and in seven subjects during an acute study. Two techniques were used for simultaneous extraction of AI and AII. Despite appropriate correction for cross reaction of AI with the AII antibody, one of the techniques gave consistently higher AII and lower AI levels in plasma of subjects treated with enalapril. The possibility of in vitro conversion of AI into AII-immunoreactive material during the purification or the radioimmunoassay steps should be considered. The use of the alternative technique is proposed for simultaneous processing of blood samples for AI and AII. PMID- 2983413 TI - Chronic T cell lymphocytosis with large granular lymphocytes of helper (OKT4) phenotype. AB - A 54-year-old asymptomatic male patient was followed for more than 7 y and presented a constant T cell lymphocytosis without skin involvement or bone marrow depression. No clinical or haematological aggravation was noted during this follow-up. Morphologically, the cells were large granular lymphocytes strongly positive for beta-D-glucuronidase, negative for acid phosphatase and with features of T cells on transmission and scanning electron microscopy. The immunological studies of the lymphocytes showed the following parameters: E rosettes+, mouse rosettes-, SmIg-, OKT3+, OKT4+, OKT8-, OKT6-, Ia-, TdT-, NK-, HTLV-, decreased PHA and PWM stimulation, no interleukin 2 production and failure to enhance Ig synthesis in a PWM driven system. The karyotype was normal. This case of chronic T cell lymphocytosis with large granular lymphocytes helper profile and defect of helper function, not reported in the literature, may correspond to a distinct entity in the heterogeneous group of chronic T cell disorders. PMID- 2983414 TI - Effect of cyclosporin and interleukin-2 on the restoration of in vitro immune responses to cytomegalovirus. AB - Previous studies have shown that cyclosporin (CSA) inhibits lymphoproliferation to cytomegalovirus (CMV)-infected, glutaraldehyde-fixed, and irradiated fibroblasts (CMVFFx) in vitro. Generation of cytotoxic cell activity is impaired in cultures with CSA, but the induction of suppressor cells is not. In the present studies we tested the ability of interleukin-2 (IL-2) and supernatants of lymphocytes stimulated by CMVFFx with or without CSA (1 microgram/ml) to restore functional activities of lymphocytes from primary cultures treated or not treated with CSA. IL-2 significantly enhanced lymphoproliferation, cell-mediated cytotoxicity to CMV-infected fibroblasts (CMVF), natural killer cell activity, and the activity of cells capable of suppressing the response of fresh autologous cells to CMVFFx of cells derived from control and CSA-treated primary cultures. IL-2 was found in day-2 supernatants of control cultures but not CSA-treated cultures. Day-2 control supernatants were capable of significantly enhancing proliferation and suppressor cell activity but were less efficient at restoring cytotoxic cell function. Day-2 supernatants from CSA-treated cultures were not able to enhance lymphoproliferation or cytotoxic cell function but did induce significant levels of suppressor cell activity. The results indicate the presence of different functional mediators in the culture supernatants. The ability of IL 2 to restore lymphocyte effector functions against a clinically important virus may have important therapeutic implications in the treatment of this viral infection in immunodeficiency diseases and in the restoration of immune competence after transplantation. PMID- 2983415 TI - Prealbumin in Swedish patients with senile systemic amyloidosis and familial amyloidotic polyneuropathy. AB - A prealbumin (PA)-like protein was demonstrated in amyloid fibrils both from a patient with senile systemic amyloidosis (SSA) and from a patient in northern Sweden with familial amyloidotic polyneuropathy (FAP). The investigated properties of this protein were similar in the two types of fibrils. The protein had molecular weight, antigenic determinants, and at least one cysteinyl residue in common with the subunit of normal PA. In contrast to normal PA, it contained disulphide-linked subunits and was to some extent bound to the fibril via the cysteinyl residues. The noncovalent forces between its subunits were weaker than in normal PA. It constituted part of the previously described AScl protein of the SSA fibrils. Proteins with lower molecular weight than the PA monomer were major proteins in both SSA and FAP fibrils. These proteins had similar properties in the two kinds of fibril and may be derived from PA. PA in serum from Swedish patients with FAP and SSA was normal with regard to the isoelectric pH of the monomers and tetramers. PMID- 2983416 TI - [Angiotensin converting enzyme in the adult respiratory distress syndrome]. AB - Angiotensin-converting enzyme (ACE) is present in high concentration in pinocytotic vesicles of the inner walls of the capillary endothelial cells of the lungs. Since a lesion of these endothelial cells is considered to be a major pathogenetic factor in adult respiratory distress syndrome (ARDS), the question arises whether serum ACE could be used as a marker enzyme in the course of ARDS to assess the syndrome's development and prognosis. The authors examined 28 patients with ARDS, 14 of whom survived acute respiratory failure. The serum ACE was determined every 48 hours. In addition, daily determination was carried out of AaDO2, pulmonary shunt perfusion (Qs/Qt) and thoraco-pulmonary compliance. In 9 of the surviving ARDS patients control serum ACE levels and lung function tests were performed half a year after recovery. 60 healthy volunteers served as controls. Definitely reduced serum ACE levels were found both in the surviving (17.1 +/- 6.9 U/ml) and dead (17.6 +/- 8.7 U/ml) ARDS patients. However, since the values for both groups are practically identical it can be assumed that determining the ACE level alone is probably not of prognostic value in acute respiratory failure. Patients who have survived ARDS show a significantly positive correlation between serum ACE and AaDO2, whereas in the patients who died a significantly negative correlation between serum ACE, AaDO2 and Qs/Qt was found.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983417 TI - Evidence for exposure to HTLV-III in Uganda before 1973. AB - Fifty of 75 serum samples collected in the West Nile district of Uganda between August 1972 and July 1973 contained antibodies reactive with human T-cell leukemia (lymphotropic) virus type 3 (HTLV-III; mean titer, 601), while 12 of 75 samples were positive in a similar test for HTLV type 1 (HTLV-1) antibodies (mean titer, 236). The samples were screened by enzyme-linked immunosorbent assay and positive results were confirmed by a newly developed unlabeled antibody peroxidase procedure with enhanced sensitivity for detection of antibody binding to immunoblots of HTLV-III antigen, demonstrating antibodies to proteins with molecular weights of 24,000, 41,000, and 76,000 in nearly all positive samples. Analysis of titration data indicated enhanced titers of antibody against HTLV-III and HTLV-I when coinfection occurred. The high prevalence and relatively low titers [compared to serum from patients with acquired immune deficiency syndrome (AIDS)] of antibodies recognizing HTLV-III proteins in sera from this population at a time that may predate or coincide with the appearance or spread of the AIDS agent (HTLV-III) suggest that the virus detected may have been a predecessor of HTLV-III or is HTLV-III itself but existing in a population acclimated to its presence. It further suggests an African origin of HTLV-III. PMID- 2983418 TI - Location of the c-yes gene on the human chromosome and its expression in various tissues. AB - Analysis of DNA from human embryo fibroblasts showed that ten Eco RI fragments were hybridizable with the Yamaguchi sarcoma virus oncogene (v-yes). Four of the Eco RI fragments were assigned to chromosome 18 and one to chromosome 6. There was evidence for multiple copies of yes-related genes in the human genome; however, only a single RNA species, 4.8 kilobases in length, was related to yes in various cells. PMID- 2983419 TI - Subcellular localization of the product of the long open reading frame of human T cell leukemia virus type I. AB - Human T-cell leukemia virus type I (HTLV-I) is a retrovirus associated with adult T-cell leukemia and lymphoma. In addition to containing the gag, pol, and env genes of the chronic leukemia viruses, the genome of HTLV-I contains a long open reading frame (LOR) located between the 3' end of the envelope gene and the 3' long terminal repeat sequence (LTR). It has been suggested that a protein of 42 kilodaltons that is encoded by the LOR region may participate in both trans acting transcriptional regulation of the viral LTR as well as in the transforming properties of HTLV-I. It is reported here that a significant fraction of the 42 kilodalton HTLV LOR product is located in the nucleus of HTLV-I-infected transformed lymphocytes, a finding that is consistent with its proposed functions. PMID- 2983420 TI - A second nuclear protein is encoded by Epstein-Barr virus in latent infection. AB - A region of the Epstein-Barr virus (EBV) genome that is important in inducing cell proliferation includes a single long open reading frame. Part of this open reading frame has been fused to the lacZ gene and expressed in Escherichia coli. Antisera to the fusion protein identify a protein in the nuclei of latently infected growth-transformed lymphocytes and in Burkitt tumor cells grown in vitro. This nuclear protein is encoded by a different virus-gene than that which encodes the previously described EBV nuclear antigen, EBNA. PMID- 2983421 TI - Antiviral chemotherapy and chemoprophylaxis. AB - Antiviral compounds have been developed for use in chemoprophylaxis and chemotherapy of a variety of infections in humans, including those caused by influenza viruses, respiratory syncytial virus, and herpesviruses. The efficacy of several of these compounds has been demonstrated in rigorously controlled trials. Advances in molecular virology have led to the identification of biochemically defined, virus-specific functions that serve as appropriate targets for the future development of antiviral compounds. Clinical investigators and practicing physicians are now confronting questions previously raised with the use of antibacterial antibiotics. These questions concern appropriate routes of administration for antiviral compounds, optimal dosage regimens, risks of long term prophylaxis, and the emergence of resistant organisms. PMID- 2983422 TI - Interaction of calcitonin and calcitonin gene-related peptide at receptor sites in target tissues. AB - Discrete receptor sites for calcitonin (CT) and calcitonin gene-related peptide (CGRP) were found in the nervous system and in peripheral tissues. Each peptide was capable of cross-reacting with the specific receptor of the other. In contrast to CT receptors, CGRP receptors were not linked to adenylate cyclase. However, CGRP could stimulate adenylate cyclase in CT target tissues apparently by interacting with CT receptors. The relative abilities of CGRP and mammalian CT to inhibit CT binding suggest that CGRP could serve as an endogenous ligand for CT receptors in the central nervous system. PMID- 2983423 TI - Measuring gene expression with light. AB - Light is produced by recombinant Escherichia coli that contain lux genes cloned from the marine bacterium Vibrio fischeri. The bioluminescence phenotype requires genes for regulatory and biochemical functions, the latter encoded by five lux genes contained in a single operon. These lux genes were disconnected from their native promoter and inserted into the transposon mini-Mu. The resulting transposon, mini-Mulux, could induce mutations by insertional inactivation of a target gene, and the lux DNA was oriented to align target gene transcription with that of the lux genes. Genes in Escherichia coli and Vibrio parahaemolyticus were mutagenized, and mutants containing transposon-generated lux gene fusions produced light as a function of target gene transcription. Light production offers a simple, sensitive, in vivo indicator of gene expression. PMID- 2983424 TI - Suppression of gamma interferon production by inactivated feline leukemia virus. AB - Supernatants from cultures of normal feline lymphocytes stimulated with Staphylococcus enterotoxin A showed antiviral activity, characterized as a gamma like interferon. With the addition of inactivated feline leukemia virus, markedly less interferon was produced. The reduction in interferon production was not attributable to lowered lymphocyte viability or reduced mitogenic properties of Staphylococcus enterotoxin A and appears to be a direct retroviral effect. This finding may reflect clinically relevant events that may contribute to the development of the feline or human states of acquired immunodeficiency. PMID- 2983425 TI - Genetic basis for species vulnerability in the cheetah. AB - A population genetic survey of over 200 structural loci previously revealed that the South African cheetah (Acinonyx jubatus jubatus) has an extreme paucity of genetic variability, probably as a consequence of a severe population bottleneck in its recent past. The genetic monomorphism of the species is here extended to the major histocompatibility complex, since 14 reciprocal skin grafts between unrelated cheetahs were accepted. The apparent consequences of such genetic uniformity to the species include (i) great difficulty in captive breeding, (ii) a high degree of juvenile mortality in captivity and in the wild, and (iii) a high frequency of spermatozoal abnormalities in ejaculates. The species vulnerability of the cheetah was demonstrated by an epizootic of coronavirus associated feline infectious peritonitis in an Oregon breeding colony in 1983. Exposure and spread of the coronavirus, which has a very low morbidity in domestic cats (approximately 1 percent), has decimated a heretofore productive and healthy captive population. The extreme genetic monomorphism, especially at the major histocompatibility complex, and the apparent hypersensitivity of the cheetah to a viral pathogen may be related, and provide a biological basis for understanding the adaptive significance of abundant genetic variation in outbred mammalian species. PMID- 2983427 TI - A virus by any other name . . . PMID- 2983426 TI - Rapid and sensitive protein similarity searches. AB - An algorithm was developed which facilitates the search for similarities between newly determined amino acid sequences and sequences already available in databases. Because of the algorithm's efficiency on many microcomputers, sensitive protein database searches may now become a routine procedure for molecular biologists. The method efficiently identifies regions of similar sequence and then scores the aligned identical and differing residues in those regions by means of an amino acid replacability matrix. This matrix increases sensitivity by giving high scores to those amino acid replacements which occur frequently in evolution. The algorithm has been implemented in a computer program designed to search protein databases very rapidly. For example, comparison of a 200-amino-acid sequence to the 500,000 residues in the National Biomedical Research Foundation library would take less than 2 minutes on a minicomputer, and less than 10 minutes on a microcomputer (IBM PC). PMID- 2983428 TI - Protection from genital herpes simplex virus type 2 infection by vaccination with cloned type 1 glycoprotein D. AB - Guinea pigs were vaccinated with truncated herpes simplex virus type-1 (HSV-1) glycoprotein D produced in the genetically engineered mammalian cell line gD10.2. Vaccinated animals formed antibodies that neutralized both HSV-1 and herpes simplex virus type 2 (HSV-2) in an in vitro neutralization assay. Vaccinated animals were challenged with HSV-2 by intravaginal infection. Animals that received the immunogen in Freund's complete adjuvant were completely protected from the clinical manifestations of genital HSV-2 infection. Animals that received the immunogen incorporated in alum adjuvants were partly protected from clinical disease; the infections that did develop were significantly less severe than those that occurred in control animals injected with adjuvant alone. The results demonstrate that immunization with a purified viral protein can provide significant protection against primary genital infection by HSV-2 in guinea pigs. PMID- 2983429 TI - Expression in Escherichia coli of open reading frame gene segments of HTLV-III. AB - Human T-cell lymphotropic virus type III (HTLV-III), the causative agent of the acquired immune deficiency syndrome (AIDS), was recently isolated and its genomic structure analyzed by DNA cloning methods. In the studies reported here a combined cloning and expression system was used to identify HTLV-III encoded peptides that react immunologically with antibodies in sera from AIDS patients. Cloned HTLV-III DNA was sheared into approximately 500-base-pair fragments and inserted into an "open reading frame" expression vector, pMR100. The inserted DNA was expressed in Escherichia coli transformants as a polypeptide fused to the lambda CI protein at its amino terminus and to beta-galactosidase at its carboxyl terminus. Sera from AIDS patients containing antibodies to HTLV-III were then used to screen for immunoreactive fusion proteins. Twenty clones, each specifying a fusion protein strongly reactive with AIDS serum, were identified. DNA sequence analysis indicated that the HTLV-III fragments were derived from the open reading frame DNA segments corresponding to the gag and pol gene coding regions and also the large open reading frame region (env-lor) located near the 3' end of the viral genome. PMID- 2983430 TI - Making antibodies without the antigens. PMID- 2983431 TI - A region of the Herpesvirus saimiri genome required for oncogenicity. AB - Herpesvirus saimiri naturally infects squirrel monkeys (Saimiri sciureus) without producing signs of disease; infection of other New World primates, however, results in a rapidly progressing, malignant, T-cell lymphoma. Results described in this report identify a region of the viral genome that is required for oncogenicity in owl monkeys (Aotus trivirgatus); this region is not required for replication of the virus. This is believed to be the first such genomic region identified in a herpesvirus system. PMID- 2983432 TI - In vivo studies on the binding of heparin and its fractions with platelet factor 4. AB - PF4 has a half-life in plasma of less than 3 minutes, and its rapid clearance appears to be a function of binding to the vascular endothelium. Once bound to the endothelium, PF4 can be released by heparin in a time-dependent manner; recovery is greater the sooner heparin is administered following PF4 infusion. This heparin-induced release of PF4 can be abolished if the heparin is first complexed with hexadimethrine bromide. Likewise, this heparin-induced release of PF4 is dependent upon the type of heparin used; low molecular weight heparin fractions and fragments do not cause the PF4 rebound seen with intact heparin. Thus, it would appear that low molecular weight forms of heparin are advantageous in that their in vivo administration would not be mediated by such platelet modulators as PF4. PMID- 2983433 TI - The role of VP-16 in the treatment of small-cell lung cancer: studies of the West of Scotland Lung Cancer Group. AB - Reviews of published studies indicate that the incorporation of VP-16 (Vepesid) into combination chemotherapy for small-cell lung cancer may improve overall response rates from 50% to between 65% and 80%. In addition, high-dose VP-16 may yield a higher response rate than that obtained with conventional doses. The West of Scotland Lung Cancer Group has therefore conducted studies to examine the effects of VP-16 both in a combination regimen as induction therapy and (together with high-dose cyclophosphamide) as late intensification therapy in high dose, aimed at preventing relapse in responding patients. Response to induction treatment improved with the addition of VP-16, compared to earlier studies carried out by the group, yielding an overall response rate of 80% for patients with limited disease and 62% for those with extensive disease. Although induction therapy comprised only three courses (lasting 9 weeks), the median response duration of 9.5 months for complete responders and the median survival of 14 months for complete responders (limited disease) were in keeping with those obtained using more prolonged induction therapy. The intensification therapy with high-dose cyclophosphamide and high-dose VP-16, however, yielded no improvement in overall survival in those responding patients who received it compared with those who did not. Radiotherapy following late-dose intensification prevented local tumor recurrence but appeared to have no effect on overall survival. Resistance to VP-16 and other drugs is a possible deterrent to successful therapy in small-cell lung cancer, and it is suggested that research focus on a possible role for calcium channel blockers in circumventing drug resistance. PMID- 2983434 TI - High-dose etoposide (VP-16) in small-cell lung cancer. AB - Etoposide (VP-16) is one of the most active drugs against small-cell lung cancer. There may be a steep dose-response relationship, and we have explored the use of etoposide as a single agent in a high dose (1,200 mg/m2) without bone marrow transplantation, for patients with very bulky, extensive-stage disease. This therapy is well tolerated in patients having good performance status, with myelosuppression representing the major toxicity. Our data suggest there may truly be a steep dose-response relationship. We have continued to explore intensive induction therapy for selected very poor-prognosis patients by adding high-dose cyclophosphamide (100 mg/kg) to high-dose etoposide. This combination is also very myelotoxic, but quite similar to etoposide alone. Our current study adds cisplatin (120 mg/m2) to the high-dose cyclophosphamide-etoposide schedule in an attempt to take advantage of the synergism seen with these drugs in various other circumstances. This series of studies will give us information regarding the feasibility of intensive induction therapy and provide data for the design of phase III studies. PMID- 2983435 TI - Single-dose etoposide in combination with vincristine and doxorubicin in the treatment of small-cell lung cancer (SCLC). AB - From July 1980, 104 consecutive patients with previously untreated small-cell lung cancer (SCLC) received vincristine 1.4 mg/m2, doxorubicin (Adriamycin) 40 mg/m2, and Etoposide (VePesid) 300 mg/m2 intravenously (as a single infusion) every 3 weeks. The overall response rate (complete response plus partial response) was 58%. In 47 patients with limited disease the response rate was 66% with 21/47 (45%) complete responders. Treatment was delivered on an outpatient basis. Toxicity was mild, and in 455 rapid infusions of etoposide, there have been no adverse reactions. PMID- 2983436 TI - Alternating combination chemotherapy regimens in small-cell lung cancer. AB - Although many combination chemotherapy regimens are capable of inducing rapid tumor regression and improved survival in patients with small-cell lung cancer (SCLC), death from recurrent chemotherapy-resistant disease remains the inevitable outcome in over 90% of cases. There have been several attempts to overcome this problem of drug resistance by the administration of two or more combination regimens in various types of alternating schedules. However, this treatment strategy has not achieved the kind of success observed in other tumor types. Several factors may account for this failure. The regimens employed may have lacked sufficient non-cross resistance and relative efficacy. Furthermore, consideration of the biologic processes involved in the regulation of tumor cell growth, heterogeneity, and drug resistance suggests that the time interval between alternating combinations should be as short as possible. Data indicate that combination cyclophosphamide, doxorubicin (Adriamycin), and vincristine (CAV), and combination cisplatin and VP-16 (PVP) are highly active and not totally cross resistant in patients with SCLC. An induction regimen consisting of CAV rapidly alternating with PVP has been studied in 44 patients. Treatment was well tolerated and the regimens could be alternated at intervals of less than 3 weeks. A 95% objective response rate and 67% complete response rate was achieved. These results appear to be better than previous results with CAV or PVP used alone as induction therapy. A randomized trial is required to confirm this impression. PMID- 2983437 TI - Cholangiography and bile cytopathology in the diagnosis of biliary tract obstruction. AB - Both percutaneous and endoscopic cholangiography identify strictures, stones, and other causes of biliary obstruction. With a radiologic diagnosis, benign and malignant causes of biliary obstruction can often be differentiated, but cholangiography does not provide a pathologic diagnosis. Therefore, we recommend cytologic examination of bile obtained during cholangiography to provide pathologic evidence of malignant causes of bile duct obstruction. We present the results of the diagnostic work-up of 17 patients with biliary obstruction to demonstrate the value of examining bile for malignant cells in all cases of biliary obstruction requiring cholangiography. If bile is cytopathologically normal, the diagnostic work-up must proceed. Abnormal findings on cytologic examination of bile in this study were confirmed with a tissue diagnosis or at operation (once) for malignant neoplasm in 100% of cases (ie, there were no false positive results). PMID- 2983439 TI - Alveolar cell carcinoma: diagnostic pitfalls in evaluating the chest roentgenogram. AB - We have reported two cases of biopsy-proven alveolar cell carcinoma, both initially thought to be pulmonary edema. The purpose of this report is to stress the variations of the roentgenographic pattern in alveolar cell carcinoma and to suggest that fiberoptic bronchoscopy with biopsy should be considered when a pattern thought to be that of pulmonary edema does not resolve with appropriate therapy. PMID- 2983438 TI - Carcinoma of the duodenum. AB - Primary duodenal carcinoma is an uncommon tumor that tends to be diagnosed late in its course because of a symptom complex compatible with many benign diseases. We reviewed the cases of five patients with duodenal adenocarcinoma treated at Norfolk General Hospital from 1973 to 1983. Periampullary tumors were excluded. The most common symptom, nausea and vomiting, was present in four patients, all of whom had microcytic anemia; in three, tests showed blood in stool specimens. The upper gastrointestinal series was suggestive of carcinoma in all patients. One of the lesions was proximal to the ampulla, while the other four were in the third and fourth portions of the duodenum. Only three of the lesions could be seen on upper GI endoscopy, and in only one of the three was the biopsy specimen positive for malignancy. At laparotomy, two patients had resectable lesions, but only segmental resection was done. Three patients had unresectable disease because of liver metastases and/or involvement of the root of the small bowel mesentery. Because delay in diagnosis of duodenal carcinoma may prevent successful resection, greater awareness of the possibility of these uncommon lesions, along with aggressive diagnostic work-up, may result in a higher percentage of cures. PMID- 2983440 TI - Primary therapy for small cell lung cancer reversing the Eaton-Lambert syndrome. AB - We have described a patient with small cell cancer of the lung manifested as the Eaton-Lambert syndrome. Primary therapy proved effective in reversing the syndrome, whereas anticholinesterase agents did not. This case illustrates the value of primary therapy for the tumor-produced syndrome and the need to avoid losing time with symptomatic therapy. PMID- 2983441 TI - Extrapulmonary small cell cancer of the retroperitoneum. AB - Small cell cancer is common in the lung, but may also be found in extrapulmonary sites, given the widespread distribution of the APUD cell, from which this tumor originates. We believe this is the first report of a small cell cancer in the retroperitoneum. An initial good response of the tumor to chemotherapy was followed six months later by appearance of metastases and patient's death. This clinical behavior is usual in extrapulmonary small cell cancer. PMID- 2983442 TI - Biologic implications of steroid hormone receptors in cancers of the colon. AB - In follow-up investigations on 41 patients with primary cancer of the colon that had been assayed for the presence of steroid-binding activity, six of eight patients whose tumors showed steroid-binding activity for at least one steroid were free of disease one to three years postoperatively. In contrast, only two of 12 patients who had negative binding assay (16%) were free of disease in the same interval. The finding of steroid receptors in human colon cancers does not immediately imply therapeutic advantage. These binding activities may not be receptors, and even if they are, cytoplasmic receptors are a necessary, but not sufficient condition for hormone response. There are, however, sufficient epidemiologic, etiologic, and nutritional observations that seem to link breast and colon cancers to indicate further investigation into the biologic significance of the association of steroid-binding protein in human colon cancer. PMID- 2983443 TI - [Problems of prognosis assessment in participation of alcoholics in self-help groups]. PMID- 2983444 TI - Analysis of Ah gene locus by somatic cell hybridization: expression of Ah regulatory gene product for 2,3,7,8,-tetrachlorodibenzo-p-dioxin in mouse L-cell x mouse hepatoma cell hybrids. AB - Properties of the aryl hydrocarbon hydroxylase (AHH) enzyme system were examined in polycyclic aromatic hydrocarbon (PAH) -noninducible L-cell x PAH-inducible hepatoma (Hepa) mouse cell hybrids. In hybrids, 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) induces AHH activity. The levels of maximal TCDD-induced AHH activity in the hybrids and the Hepa parent are similar, although a greater concentration of TCDD is required for expression in the hybrids. This concentration difference appears to reflect dilution of AHH-associated gene products by the L-cell parent rather than altered gene expression. The regulatory gene product, the Ah receptor, is expressed similarly in the hybrids and Hepa parent. Both demonstrate specific, high-affinity binding of [3H]TCDD to an equivalent number of receptor sites per cell. These results suggest that the molecular mechanism of phenotypic resemblance to the inducible Hepa parent (i.e., "dominance") in the mouse L-cell x Hepa hybrids involves expression of only the Hepa Ah gene complex. PMID- 2983445 TI - Malignant stricture at the confluence of the biliary tree: diagnosis and management. PMID- 2983446 TI - Mammography in women less than 40 years of age. AB - Carcinoma of the breast in women less than 40 years old is a significant problem, representing 7.4 per cent of the total number of carcinomas of the breast at our institution. It is apparent that mammography in this age group is of value in confirming the clinical suspicion of a malignant condition of the breast and in evaluating the remainder of the parenchyma of the breast for clinically occult lesions. However, one cannot over emphasize that a negative result from the mammogram should not delay biopsy in the presence of clinically suspicious lesions (1, 4). It is obvious that a larger series of patients is necessary to define the role of mammography in "young women." PMID- 2983447 TI - Carcinoma of the breast associated with pregnancy. AB - Sixty-three pregnant patients were treated for primary carcinoma of the breast at the Mayo Clinic between 1950 and 1980. These patients had a greater incidence of nodal metastasis and scirrhous histopathologic factors than did non-pregnant patients with a malignant condition of the breast. The five year survival rate of pregnant patients with localized malignant growths of the breast was 80 per cent. Improved survival time was not demonstrated for patients who terminated the pregnancy. PMID- 2983448 TI - Prognostic value of round cell (lymphocyte) infiltration in malignant gliomas. AB - The biopsy specimens of 342 patients with malignant glioma were evaluated to determine the extent and prognostic significance of round cell infiltration within these representative tissues: (a) tumor, (b) peritumoral, (c) hypervascular, (d) necrotic, and (e) normal tissue. Thirty-six percent of all biopsy specimens showed at least one tissue area with round cell infiltration. Patients in age groups 0 to 25 years and 71 years and older tended to show less round cell infiltration than did patients 41 to 55 years old. The presence of round cell infiltration in tissue was associated with a poor prognosis. Patients showing any infiltration had mean survival times of 8.4 months as opposed to 11.9 months for those showing no infiltration. The relationship between round cell infiltration and poor prognosis is true, irrespective of postoperative therapy, sex, and age. PMID- 2983449 TI - Results of surgical treatment of radiculomyelopathy caused by cervical arthrosis based on 1000 operations. AB - Wide experience over a 20-year period with 1000 interventions for radiculomyelopathy caused by cervical arthrosis is reported. The results are analyzed according to clinical signs, surgical methods, timing of operation, influence of pathogenic phenomena, age, and preexisting illness. Intra- and postoperative problems and accidents are covered. PMID- 2983450 TI - Effect of hydroxyl radical scavenging on endotoxin-induced lung injury. AB - The release of oxygen radicals, in particular the hydroxyl radical, from sequestered neutrophils produces acute lung injury after a number of insults. Our purpose was to determine whether hydroxyl radical, OH., is responsible for the lung injury from endotoxin characterized by (1) pulmonary leukostasis, (2) increased thromboxane production leading to pulmonary hypertension and hypoxia, and (3) increased protein permeability. This hypothesis was tested by infusion of a selective OH. scavenger, dimethyl thiourea (0.75 gm/kg), into unanesthetized sheep before endotoxin and comparison of the response to that seen with endotoxin alone. Pulmonary vascular integrity was measured by the use of lung lymph flow, QL, and lymph protein transport. Thromboxane A2 was measured as TxB2 and prostacyclin as 6-keto-PGF1 alpha. We found no difference in the degree of leukopenia and hypoxia after endotoxin or the levels of TxB2, 6-keto-PGF1 alpha, and pulmonary hypertension with dimethyl thiourea, compared with endotoxin alone. The permeability injury was also identical, with a twofold to threefold increase in protein-rich lymph seen in both groups. It appears that OH. does not play a major causative role in either phase of endotoxin lung injury. PMID- 2983451 TI - Localization of an occult insulinoma by intraoperative ultrasonography. AB - Intraoperative ultrasonography has not been used previously to locate an insulinoma that was not surgically palpable or that could not be seen by selective arteriography. In this report we described a patient with an insulinoma localized to the pancreatic head identified by transhepatic portal venous sampling but not by selective arteriography or palpation. At operation intraoperative ultrasonography demonstrated an 8 mm tumor in the pancreatic head and guided the successful enucleation. PMID- 2983453 TI - Cigarette smoking does not affect PGI2 platelet receptors. PMID- 2983452 TI - Neurone specific enolase: a useful diagnostic serum marker for small cell carcinoma of the lung. AB - Among lung cancers small cell carcinoma is the most sensitive to chemotherapy and radiation. This has emphasised the importance of an accurate diagnosis of this cell type, and the present study examined the use of serum neurone specific enolase (NSE) as a diagnostic marker for small cell carcinoma. NSE was measured in pretreatment sera from 103 patients with small cell carcinoma and in sera from relevant controls, including patients with other lung cancers, non-malignant lung diseases, and healthy adults. Serum NSE concentration was raised (greater than 25 ng/ml) in 72% of patients with small cell carcinoma. Ninety one per cent of patients with extensive disease and 50% of patients with limited disease were serum NSE positive. Patients with extensive disease in general had higher serum NSE concentrations than patients with limited disease. No definite difference in serum NSE positivity could be shown between oat cell and intermediate cell subtypes. Out of 51 patients with other lung cancers, four (8%) had a raised serum concentration, whereas all patients with non-malignant diseases and healthy individuals had normal serum NSE concentrations. Serum NSE determination seems to be a valuable tool for the diagnosis of small cell carcinoma. PMID- 2983454 TI - [Mastitis--Leptospira hardjo--milker's fever]. PMID- 2983455 TI - Dimercaptan metal-binding agents influence the biotransformation of arsenite in the rabbit. AB - The urinary metabolites of sodium arsenite have been investigated in rabbits given sodium arsenite and water-soluble dimercaptans. Rabbits injected sc with NaAsO2 (1 mg As/kg) were given, im 1 hr later, either saline, 2,3-dimercapto-1 propanesulfonic acid (DMPS), mesodimercaptosuccinic acid (DMSA), or N-(2,3 dimercaptopropyl)phthalamidic acid (DMPA) at 0.2 mmol/kg. Arsenic metabolites in urine collected from treated rabbits were isolated by combined anion-cation exchange chromatography. Column fractions were acid-digested and analyzed for arsenic by direct hydride-flame atomic absorption spectrophotometry. The relative amounts of inorganic arsenic, methylarsonate, and dimethylarsinate found in 0 to 24 hr urine of rabbits given only sodium arsenite agreed closely with those reported for human subjects given arsenite po. This finding suggests that the rabbit biotransforms arsenite in a manner very similar to that of man. The urinary excretion of total arsenic between 0 and 24 hr was elevated after dimercaptan administration, but urinary excretion of total arsenic between 0 and 48 hr was unaffected. This result indicates that the action of these dimercaptans occurs early after treatment. In addition, the dimercaptans influenced differently the amounts of the arsenic metabolites excreted in the urine between 0 and 24 hr. DMPS, DMSA, or DMPA increased arsenite excretion but decreased dimethylarsinate excretion. DMPS or DMPA treatment increased methylarsonate excretion but DMSA did not. Arsenate excretion increased after DMPS or DMSA treatment but was not affected by DMPA treatment. These results suggest that the dimercaptans, in addition to increasing arsenic excretion, also influence the biotransformation of arsenite to less toxic species. The different effects on the urinary excretion of arsenic metabolites suggest that these dimercaptan metal binding agents have mechanisms of action in addition to simple chelation of inorganic arsenic. PMID- 2983456 TI - Cytotoxicity related changes in biochemical cell function following in vitro cadmium treatment. AB - In an effort to examine cellular responses to cadmium insult a bovine kidney cell line was used to monitor select cell functions for toxicity related alterations. Cadmium concentrations used ranged between 0.2 and 2.5 microM CdCl2 and elicited 0-85% cytotoxicity (cell attachment); 24-h incubations were used for all studies. Toxicity related inhibition of leucine incorporation into cellular protein and thymidine incorporation into DNA was noted. Decreases in protein synthesis activity closely paralleled the cytotoxicity profile; DNA synthesis was a less sensitive indicator to toxicity. K+-dependent phosphatase (KP), acid phosphatase (AP) and succinate dehydrogenase (SDH) were monitored in surviving cells and in a cell-free system. Significant inhibitions were detected for all enzyme activities following a 24 h culture with cadmium. KP and AP were most sensitive. In the cell free system KP was significantly inhibited with 0.1 microM cadmium; AP and SDH were either unchanged or sensitive only at concentrations of 100 microM cadmium or greater. Reduced glutathione (GSH) concentration in surviving cells was elevated up to 7-fold over control cultures. The elevation occurred in a progressive toxicity-related manner. PMID- 2983457 TI - Influenza type A virus infection of mice exposed in utero to chlordane; survival and antibody studies. AB - Previous studies carried out by others have shown that in utero exposure of mice to chlordane effects a significant depression of cell-mediated immunity (CMI) at 100 days of life without adversely affecting the humoral immune system. In the studies reported herein we assessed the effect of in utero exposure to various doses of chlordane on the response of 38-day-old mice to influenza type A virus infection in terms of relative levels of mortality, mean day of death, and the levels of antiviral antibody in the primary and secondary immune response to the virus. In utero exposure to chlordane effected enhanced survival to influenza type A virus infection relative to mock-treated animals. No significant differences were noted in the mean day of death of chlordane-treated and mock treated mice. A significant enhancement in the levels of antiviral antibody was noted in the chlordane-treated female mice but not male mice in both the primary and secondary immune response to the virus. PMID- 2983458 TI - In vivo effects of nitrogen dioxide on the blood nitrate level and the Na+,K+ ATPase activity of red blood cells of rats. AB - Male Wistar rats were exposed to 0.4, 1.2 and 4.0 ppm NO2 for 13 weeks to examine the effects of NO2 on the blood nitrate concentration and the Na+,K+-ATPase activity of red blood cells. Exposures to 1.2 and 4.0 ppm NO2 caused an elevation of the blood nitrate level at the first, third and eighth week. The maximum concentration attained was 148% (P less than 0.001, at the third week) and 201% (P less than 0.001, at the eighth week) of the controls at 1.2 and 4.0 ppm NO2, respectively. On the other hand, the nitrate concentration was decreased to the control level at the second, fourth and thirteenth weeks. 0.4 ppm NO2 caused a progressive but slight increase in the blood nitrate concentration from the third week and reached the maximum (122% of the control, P less than 0.001) at the eighth week. The Na+,K+-ATPase activity decreased slightly from the first week upon exposure to 4.0 ppm NO2 and reached the minimum (72% of the control, P less than 0.05) at the third week. Subsequently, the activity was increased to 159% (P less than 0.001) of the control at the eighth week. Exposure to 0.4 and 1.2 ppm NO2 caused fundamentally similar but less significant alterations of the Na+,K+ ATPase activity. PMID- 2983459 TI - [Treatment of non-small-cell bronchial carcinoma with cisplatin, ifosfamide, vindesine and VP 16]. AB - Hitherto, the objective of chemotherapy in case of the non-small cell bronchial carcinoma is only of a palliative nature. Thus a critical indication is necessary. This is confirmed by our investigations with the combinations of cis platinum and ifosfamide (80 patients, remission rate 35%, median survival time of patients with remission 11,5 months), cis-platinum and vindesine (29 patients, remission rate 28%, median survival time of patients with remission 14,5 months), and ifosfamide and vepeside (63 patients, remission rate 27%, median survival time of patients with remission 12 months). The combination ifosfamide-vepeside was much better tolerated by the patients and, with its comparable remission rates and survival times, was superior to the cis-platine combinations. For the chemotherapy of the non-small cell bronchial carcinoma it has to be considered that the treatment result may be more influenced by tumor stage and activity index of the patient than by the therapy method. PMID- 2983460 TI - Constricting effect of leukotrienes on cerebral arterioles of mice. AB - Three leukotrienes (LTB4, C4, D4) were locally applied to cerebral arterioles in doses varying from 3 X 10(-8) to 4 X 10(-7) M and each produced a dose related constriction. LTC was tested in the presence and absence of FPL-55712, a relatively specific inhibitor of leukotriene receptors. A low dose (10(-8) M) of FPL-55712 did markedly reduce the response to the LTC and did not significantly alter an equally pronounced constriction produced by norepinephrine. The constricting properties of the leukotrienes shown by our data support the suggestion that these products of lipoxygenase's action on arachidonic acid have the potential to modify cerebrovascular tone. This potential may be important in view of recent evidence showing that the brain can make significant amounts of leukotrienes from arachidonic acid. PMID- 2983462 TI - Transmission of latent cytomegalovirus in a murine kidney tissue transplantation model. AB - This paper examines the hypothesis that latent murine cytomegalovirus (MCMV) may be transmitted in kidney tissue to transplant recipients. Balb/c mice were infected with MCMV, and at intervals of less than 1 week to greater than 1 year, transmission of the virus from infected donors was attempted by transplantation of kidney sections or transfusion of blood into uninfected recipients. Graft recipients were killed from 2-4 weeks later, and cultured for MCMV. Restriction endonuclease digestion profiles of viral DNA were performed. Acutely infected donors transmitted MCMV in kidney tissue to 83-66% of untreated, susceptible recipients. Latently infected donors transmitted the infecting strain of virus to 20% of all and 31% of immunosuppressed recipients but to 37% of the syngeneic versus 21% of the allogeneic (P less than .027). Blood transfusions transmitted latent virus to 28% of recipients. In conclusion, kidney tissue may serve as the source of latent virus in this murine transplantation model. Retained blood in the kidney is unlikely to be the only source of virus. PMID- 2983461 TI - Prophylactic oral acyclovir after renal transplantation. AB - In a double-blind, controlled study 35 herpes simplex virus (HSV) antibody positive patients were randomized to receive oral acyclovir 200 mg X 4 daily or placebo for 28 days following renal transplantation. The incidence of herpes virus infection was compared in both groups by weekly virus demonstration/isolation testing from throat swabs and urine, and by serum antibody demonstration. None of the 18 patients allocated to acyclovir showed any signs of HSV or varicella zoster virus (VZV) infection during the trial period, whereas 9 of 17 receiving placebo had signs of HSV (P less than 0.001) and 2 of VZV (P less than 0.05) infection. Because of systemic as well as local symptoms of infection in five of the placebo patients, the trial was interrupted and treatment with oral acyclovir instituted. All of them responded well with rapid disappearance of all symptoms. Cytomegalovirus (CMV) was isolated from the urine of two patients in both groups during the trial period; a significant antibody rise was seen later in three of them. There was no evidence of drug-related toxicity during the study. PMID- 2983463 TI - Percoll separation of leukemic leukocytes for MLC matching prior to bone marrow transplantation. PMID- 2983464 TI - Acquired immunodeficiency syndrome (AIDS) in Africa. A review. PMID- 2983465 TI - [Effect of the permeability of lysosomal and mitochondrial membranes on the ability of their cryoextracts to inhibit the protein-synthesizing activity of cell-free systems]. AB - Changes in permeability of lysosomal and mitochondrial membranes were studied under different temperatures at which cryoextracts from organelles inhibit most distinctly the protein synthesizing activity of the cell-free system from the rat liver. It is found that mitochondria are more sensitive to the effect of low temperature effect than lysosomes. Overcooling of the mitochondria suspension to the temperature of the free water crystallization is shown to cause no release of the protein synthesis inhibitor. The inhibitor release from organelles occurs from the moment of crystallization and reaches its maximum at the eutectic temperatures of the freezing medium which is due to the injury effect of the complex of physicochemical factors on the membrane structures, occurring during the phase transition of the solvent. PMID- 2983466 TI - [Phosphorylase and glucose-6-phosphatase activity in rat tissues during ontogenesis]. AB - The activity of phosphorylase and glucose-6-phosphatase was determined in the liver, cerebral hemispheres, musculus gastrocnemius and myocardium of uneven-aged rats. The phosphorylase activity was the highest in rats aged 14-30 days and the glucose-6-phosphatase activity--in rats aged one day. Considerable age changes are observed in the ratio of phosphorylases a and b. PMID- 2983467 TI - [Effect of freezing and thawing on the structure and function of cytochrome oxidase]. AB - The effect of freezing-thawing on cytochrome oxidase from cattle heart was studied. The enzyme was characterized for its activity, absorption spectra, temperature perturbation differential spectra and conformational transitions. A decrease in the activity and conformational changes depend on the composition of the buffer system and the presence of potassium and sodium chlorides in the solution. PMID- 2983469 TI - [AIDS and human T-cell leukemia/lymphoma virus III in Denmark]. PMID- 2983468 TI - [A new method correcting metabolic acidosis]. AB - A new method is suggested for correction of metabolic acidosis. The method consists in a decrease of pH of the bicarbonate sodium solution used for intravenous injection down to the physiological value of pH in the blood at the expense of buffering the solution by CO2. Experimental data conforming the efficiency of the new method application are presented. PMID- 2983470 TI - Prostatic malacoplakia. AB - We report here a 70-year-old man with prostatic malacoplakia which is an extremely rare disease. This diagnosis was obtained from histological examination by prostatic biopsy because the patient was suspected having prostatic cancer on digital examination. PMID- 2983471 TI - [Morphologic and cell kinetic studies of prostate cancers. Contribution to grading]. AB - The combined histologic and cytologic grading of carcinomas of the prostate is not only important in evaluation of prognosis but determines the choice of therapy with knowledge of staging. Tissue biopsies of 2,200 prostatic carcinomas were classified and graded histologically and cytologically. Furthermore, autoradiographic studies were performed on 69 needle biopsies with 3H-thymidine. The cytologic parameters were correlated with cell-kinetic parameters in classification. Several subgroups of degrees of malignancy were found. Grade Ia corresponds to highly differentiated glandular carcinomas by histology and cytology. Grade Ib carcinomas were histologically well but cytologically moderately differentiated. Grade IIa carcinomas are histologically moderately to poorly but cytologically moderately differentiated. Grade IIb carcinomas correspond to poorly differentiated tumors by histological and cytological criteria. Grade III carcinomas are in most cases undifferentiated. This differentiated grading is particularly important for therapeutic consequences in incidental carcinomas. The therapeutic consequences of grade Ia carcinomas are frequent controls and a wait-and-see attitude. Grade Ib and IIa carcinomas must be treated according to their clinical stage by total prostatectomy. Grade IIb and III carcinomas need a palliative treatment by hormones, castration, irradiation or cytostatic drugs. PMID- 2983472 TI - Diagnostics and therapy of germinal testicular tumors. AB - Based on experience in the treatment of 627 patients with germinal testicular tumor and referring to recent literature, the age distribution (children, 3.6%, men over 50 years, 6.3%), importance and possibilities of early detection as well as sensitivity, specificity and accuracy of tumor markers, ultrasonography and X ray examinations are described. In N0M0 stage seminomas, cure can be effected by radiotherapy in almost 100% of the cases. An alternative 'watch policy' is discussed. In N1-2M0 stage seminomas, cure can be achieved by irradiation in more than 90% of the cases. Primary polychemotherapy is needed in stage N3M0 or M1 as well as in stage N4M0 and N1-4M1 seminomas. Complete remission can be obtained in more than 90% of the patients if salvage operation, further chemotherapy or radiotherapy is performed in cases without complete remission after semicastration and primary chemotherapy. In N0M0 stage non-seminomas (excluding pT4 cases, choriocarcinoma, and patients with persistently elevated markers following semicastration), 'watch policy' has the disadvantage of requiring optimal monthly follow-ups and progression in 20% of the cases. While modified lymphadenectomy reduces the progression rate to 10% with low operative morbidity, it leads to an irreversible loss of ejaculation in 12% of the patients. With both modalities, if progression is detected, full recovery can be expected with immediate polychemotherapy. In stage N1-2M0 non-seminomas, a tumor-free condition can be obtained in close to 100% of the cases by lymphadenectomy and adjuvant chemotherapy. In stage N3,4 and/or M1, first primary polychemotherapy is carried out. In the case of a residual tumor this is followed by salvage operation and, if active tumor is found, by salvage chemotherapy. With this treatment, recovery can be achieved in 70-80% of the cases depending on the involvement of surrounding organs. PMID- 2983473 TI - Pediatric urologic radiology. Intervention and endourology. AB - Over the past 10 years new imaging and interventional techniques have drastically changed the ease and scope of urologic diagnosis and treatment. It is both rewarding and exciting to approach each clinical problem with a broad armamentarium of available studies, always seeking the most efficient and direct route to diagnosis. Similarly, radiologic interventional techniques are potentially applicable to a multitude of problems and should be innovatively considered in the urologic patient including patients in the pediatric age group. PMID- 2983475 TI - Urachal adenocarcinoma presenting as vesicoenteric fistula. AB - We report a case, which we believe to be the first, of urachal carcinoma presenting as vesicoenteric fistula. PMID- 2983474 TI - Pediatric urologic oncology. AB - Effective chemotherapy combined with surgery has produced excellent disease-free survival rates in most children with genitourinary malignancy. For patients with Wilms' tumor, the overall 2-year survival rate with no evidence of disease is greater than 90 per cent, and the interval of chemotherapy continues to be reduced for those children with low-stage lesions and favorable histologic patterns. In children with pelvic rhabdomyosarcoma, primary cytoreductive chemotherapy has proved to be an effective alternative to exenterative surgery. Patients with yolk-sac tumor who have localized disease need not be subjected to the potential morbidity of retroperitoneal lymph-adenectomy or the toxicity of adjuvant chemotherapy. Conversely, patients with advanced disease can be salvaged with recently developed combination chemotherapeutic regimens. PMID- 2983476 TI - Wilms tumor arising in horseshoe kidney. AB - Tumors originating in horseshoe kidneys are rare; only 21 cases of Wilms tumor reported in horseshoe kidneys. A seven-month-old girl with Wilms tumor arising from the isthmus of a horseshoe kidney is reported. The literature is reviewed, and surgical considerations are discussed. PMID- 2983477 TI - Staging of renal cell carcinoma: cost-effectiveness of routine preoperative bone scans. AB - The use of bone scans in the evaluation of renal cell carcinoma has become routine in many centers. In a retrospective analysis of 42 patients undergoing radical nephrectomy for renal cell carcinoma, we analyzed the cost-effectiveness of routine preoperative bone scans. Although these scans accurately predict metastatic disease to bone, they are not cost-effective as a routine preoperative tool because they do not alter outcome. In selected patients with bone pain and no other positive staging studies, preoperative bone scans may be of value in the decision to perform extirpative surgery. PMID- 2983478 TI - Pathogenesis of canine parvovirus enteritis: the importance of viremia. AB - The clinical signs, hematologic changes, serum and fecal virus titers, specific antibody production and the occurrence of histologic lesions were studied in 22 nine-week-old seronegative beagle dogs inoculated by the oral and intravenous route with canine parvovirus. Approximately 30% of the dogs had clinical signs of pyrexia, depression, vomiting, and diarrhea irrespective of the route of inoculation. Events in the dogs inoculated intravenously preceded those in dogs inoculated orally by approximately two days. Only one dog died. Lymphopenia was the most consistent hematologic change. Viremia always preceded the initiation of fecal virus shedding. Viral titers in the serum and feces were significantly greater in symptomatic dogs compared to asymptomatic dogs. Termination of the plasma viremia coincided with the onset of the humoral immune response, but viremia persisted one day longer in symptomatic dogs. The severity of lymphoid tissue and intestinal infection, assessed by tissue immunofluorescence and histology, was also greater in symptomatic dogs. The severity of intestinal disease was highly correlated with the magnitude and duration of viremia. PMID- 2983479 TI - Cystic nephroblastoma of an aged dog. PMID- 2983480 TI - Application of the haemagglutination inhibition test to typing of infectious bronchitis virus. PMID- 2983481 TI - Prevalence of antibody to typical and atypical rotaviruses in pigs. PMID- 2983482 TI - Bovine respiratory disease. PMID- 2983484 TI - Outbreak of balanoposthitis in breeding bulls. PMID- 2983483 TI - Infection of lambs with caprine arthritis encephalitis virus by feeding milk from infected goats. PMID- 2983485 TI - Eicosanoids in health and in disease: an appraisal. AB - Oxygenation of the 20-carbon polyunsaturated fatty acid, arachidonic acid, which is found in most body cells of all domestic animals, leads to the formation of a group of compounds possessing biological activity. These compounds, collectively known was eicosanoids, currently receive considerable attention owing to their involvement in a wide variety of physiological and pathophysiological processes. Particular interest has been focussed in recent years on the role and control of prostanoids and leukotrienes in inflammatory and allergic conditions in animals and man. Arachidonic acid metabolites are also recognised to be intimately involved in reproductive and perinatal processes; with platelet aggregation and vascular homeostasis; kidney function; fever; certain tumours and many other normal and disease conditions. Eicosanoid research in veterinary medicine is still at a relatively early stage in many respects and in this review an attempt is given to highlight some of the functions of this important series of compounds both in health and in disease. As more evidence comes to light, it is possible that veterinary surgeons may have to consider revising their clinical approach to the treatment of certain disease states where eicosanoids are implicated or where chemotherapy may interfere with their normal physiological activities. PMID- 2983486 TI - Paramyxovirus disease in racing pigeons. Clinical aspects and immunization. A report from the Netherlands. AB - Since 1981 a highly contagious viral disease causing high morbidity and low mortality in racing pigeons has spread over Europe. The virus belongs to the avian paramyxovirus sero group I. Clinical signs include watery droppings, polydypsia and neurologic signs in a high proportion of infected animals. Definitive diagnosis can be made by virus isolation in cell cultures or chicken embryos, and virus identification by haemagglutination and haemagglutination inhibition (HI) tests. The HI test, using sera from suspected animals, is a useful clinical tool to confirm the diagnosis. The most important differential diagnosis is salmonellosis. Good immunity against this disease can be acquired by subcutaneous vaccination with an inactivated oil adjuvant poultry NDV-vaccine. For the benefit of pigeon racing a plea is made for compulsory vaccination in countries in which the disease is endemic. PMID- 2983487 TI - Existence of cytotoxic activity against BLV-transformed cells in lymphocytes from normal cattle and sheep. AB - Peripheral blood lymphocytes (PBL) from normal cattle and sheep were tested for their cytotoxic activity against several target cells using a 20-hour 51Cr release assay. The following characteristics of the effector cells were observed; 1) PBL from animals showed cytotoxic activity against two sheep cell lines (FLK and SF-28) that were transformed with bovine leukemia virus. However, normal sheep and bovine cells and Molony leukemia virus-induced mouse lymphoma cell line (YAC-1) were not killed by these cells. 2) A time course study showed that the activity was first observed at 4 to 8 hours and reached a maximum at 20 to 30 hours after incubation. 3) Cytotoxic activity was observed in both adherent and nonadherent cell fractions when PBL were passed through a nylon-wool column. This indicated that the effector cells showed some degree of adherence. 4) Treatment of PBL with carrageenan did not change the cytotoxic activity against target cells, indicating that phagocytic capability is not perhaps necessary for cytotoxicity to take place. These results indicate that the effector cells participating in the cytotoxic reaction resembled natural killer cells or natural cytotoxic cells which are present in murine and human systems. However, analysis of the cell surface markers of the effector cells is yet to be done in future studies. PMID- 2983488 TI - Indirect immunofluorescence and immunodiffusion tests in the detection of antibodies to foot-and-mouth disease virus. AB - The antibody response detected by indirect immunofluorescence (IIF) as well as that directed against 140 S and virus infection associated antigen (VIA), as detected by agar immunodiffusion, was studied in three mammal species susceptible to Foot and Mouth Disease Virus, after challenge with living virus, immunization and hyperimmunization with inactivated virus, and immunization followed by challenge. By spot indirect immunofluorescence, antibodies were detected only in animals undergoing an active infection, and were not detected in immunized or hyperimmunized animals. This behaviour was similar to that of the anti-VIA antibodies in the same groups of animals and differed from that of anti-140 S antibodies. It appeared that spot indirect immunofluorescence for the detection of VIA antigen is comparable to the immunodiffusion test, but the speed of IIF and the possibility of handling many samples make it more practical. PMID- 2983489 TI - Immunohistochemical distribution of S-100 protein and glial fibrillary acidic protein in normal and neoplastic salivary glands. AB - Immunohistochemical localization of S-100 protein, its alpha and beta subunits, and glial fibrillary acidic protein (GFAP) in normal and neoplastic salivary glands was studied by the peroxidase-antiperoxidase method and immunoblot analysis. Positive immunostaining for S-100 protein was observed in pleomorphic adenoma, adenolymphoma, tubular adenoma, adenoid cystic carcinoma, acinic cell tumour, adenocarcinoma and carcinoma in pleomorphic adenoma. S-100 protein was localized in myoepithelial cells, epithelial cells of intercalated ducts and serous acinar cells of normal salivary gland. Both alpha and beta subunits of S 100 protein showed almost identical distribution in normal and neoplastic salivary glands, but skeletal muscle cells were alpha-positive/beta-negative whereas Schwann cells and fat cells were alpha-negative/beta-positive in the stroma and neighbouring tissue. GFAP was only found in pleomorphic adenoma and its malignant counterpart. Immunoblot analysis showed that the GFAP-related antigen consisted of several polypeptide bands with a molecular weight ranging between 35,000 to 50,000 daltons. PMID- 2983491 TI - Mapping of Epstein-Barr virus proteins on the genome by translation of hybrid selected RNA from induced P3HR1 cells and induced Raji cells. AB - RNA was isolated from induced P3HR1 cells which synthesize Epstein-Barr virus (EBV) particles and therefore a full set of early and late antigens and from induced Raji cells which synthesize only early EBV proteins and hybridized to cloned EBV-DNA fragments spanning the entire genome. Bound mRNA was eluted and translated in vitro with rabbit reticulocyte lysate. The translation products were analyzed on SDS-polyacrylamide gels either directly or after immunoprecipitation with human sera. Most proteins could be mapped to short defined regions of the EBV genome using short restriction fragments and overlapping sheared fragments and there is evidence of splicing for some mRNA species. The synthesis of five early proteins can be seen only with hybrid selected RNA from induced Raji cells. These mRNAs seem to be enriched in the cells restricted to early antigen synthesis. PMID- 2983490 TI - Tumour volume and macroscopic growth pattern of bronchogenic carcinoma. AB - 126 resected malignant lung tumours were cut into serial sections. The tumour volume, the pTN-stage and the macroscopic growth pattern were computed. The average tumour volume was found to be 28.5-35.8 cm3 (confidence limits, p greater than or equal to 95%) and to be independent of the major cell type of the carcinomas. The cell types grow in different macroscopically distinguishable tumour growth patterns: epidermoid carcinomas grow mostly in bizarre finger-like shapes and small cell anaplastic and adenocarcinomas more frequently in ball-like shapes. The different growth patterns depend on the cell type, the cellular response of the host tissue and the tumour volume. Due to irregular growth pattern the tumour volume computed by X-ray diameter is likely to be overestimated by 50%-300%. PMID- 2983492 TI - Polytranscripts of Sendai virus do not contain intervening polyadenylate sequences. AB - Discrete high-molecular-weight RNA species with the properties of polytranscripts were observed in poly(A)-rich RNA extracted from Sendai virus-infected cells. These RNA species were virus specific, being synthesized in the presence of actinomycin D, but not seen in uninfected cells. They were not genome or antigenome fragments, since they were not encapsidated, as shown by their destruction when ribonuclease was added to cell homogenates and by their absence from the RNA fractions that did not bind to oligo(dT)-cellulose. Two lines of evidence indicated that the gene-specific regions of these polytranscripts were not linked by poly(A) sequences, but were faithful copies of virus genomic RNA sequences at gene boundaries. First, a small cDNA clone obtained by reverse transcription of poly(A)-rich RNA species from infected cells contained 90 bases from the 5' terminus of the gene for the P protein and about 600 bases from the 3' end of the downstream gene, which specifies the M protein, the entire cloned sequence being an accurate complement of the genomic RNA. Second, dideoxynucleotide sequencing of poly(A)-rich RNA species primed by virus gene specific oligodeoxynucleotides revealed read-through products of transcription containing no detectable poly(A). If Sendai virus polytranscripts are intermediates in the production of monocistronic viral mRNAs by a cleavage process, and poly(A) sequences do not link the mRNAs, polyadenylation would have to follow the cleavage step; it seems more likely that these polytranscipts are aberrant transcription products generated by occasional termination failure in a stop-start mechanism of transcription. PMID- 2983493 TI - Use of a focal immunofluorescence assay on live cells for quantitation of retroviruses: distinction of host range classes in virus mixtures and biological cloning of dual-tropic murine leukemia viruses. AB - A rapid and sensitive focal immunofluorescence assay (FIA) using monoclonal antibodies or heterologous antisera was employed for detection and biological cloning of viruses capable of inducing viral antigens on cell surfaces. The FIA was performed directly on a variety of live cells in tissue culture dishes and was used successfully with C-type murine leukemia viruses (MuLV) of different tropism including ecotropic, xenotropic, amphotropic, and dual-tropic recombinant mink cell focus-inducing (MCF) viruses. With the FIA, we were able to titrate and distinguish ecotropic Friend-MuLV and Friend-MCF viruses present in mixtures. Dual-tropic MCF viruses could be specifically detected directly in mouse cells by using MCF-specific monoclonal antibodies. These antibodies replaced the requirement for production of typical MCF cytopathic effect in mink cells for MCF virus detection, and also allowed efficient titration in mouse cells of MCF virions pseudotyped with ecotropic envelope proteins. Furthermore, by picking foci of fluorescent cells and using their cell-free viral progeny, MCF viruses were cloned from complex pseudotypic mixtures. This allowed the cloning of viruses present at low frequency in heterogeneous mixtures obtained from leukemic tissues. PMID- 2983494 TI - Different murine cell lines manifest unique patterns of interference to superinfection by murine leukemia viruses. AB - Interference to superinfection by murine leukemia viruses (MuLV) was analyzed in cells chronically infected with other MuLVs. A new sensitive focal immunofluorescence assay employing monoclonal antibodies was used to detect foci of virus infection in live cell monolayers. Monoclonal antibodies were chosen which reacted with the challenge virus but not with the interfering virus. The results obtained confirmed some of the findings of previous workers using Moloney sarcoma virus pseudotypes as challenge viruses on mouse and nonmouse cells. In addition, SC-1 mouse cells nonproductively infected with defective spleen focus forming virus were found to be resistant to superinfection by recombinant dual tropic viruses. Furthermore, results indicated that interference patterns between some pairs of viruses differed in different cell types. Thus, xenotropic MuLV blocked superinfection by recombinant dual-tropic viruses in SC-1 feral mouse cells, but not in two lines of NZB mouse cells. Also, in a Mus dunii tail fibroblast cell line some unique patterns of interference were observed. One ecotropic MuLV blocked infection by two xenotropic viruses and three recombinant dual-tropic viruses. Two other ecotropic viruses blocked infection by only one of the two xenotropic viruses tested. These two ecotropic viruses also differed from each other in their ability to block the three recombinant viruses. In addition, two strains of amphotropic MuLV also differed in their interference capacity. As expected, strain 1504A did not block any viruses tested, whereas strain 4070A surprisingly blocked one xenotropic and one ecotropic MuLV. The lack of homogeneity in interference patterns seen in the Mus dunii cells suggested either that a large number of heterogeneous virus receptors were present on this cell line or that interference in these cells might operate through a mechanism other than blocking of virus receptors by the envelope protein of the interfering virus. PMID- 2983495 TI - Quantitative analysis of visna virus replication in vivo. AB - Visna virus is the prototype of the lentivirus subfamily, a group of nontransforming retroviruses that cause slow infections in sheep and goats. In nature, virus is acquired primarily by the respiratory route and subsequently spreads to several organ systems. These viruses persist for years in their hosts despite a vigorous immune response because of a block in virus gene expression. This report continues the analysis of persistence in vivo, and specifically examines a gene dosage hypothesis that has been advanced as an explanation for the decrease in transcription and virus production in the cells in infected animals. For this analysis a new pulmonary model has been developed that, in conjunction with quantitative in situ hybridization, provides an opportunity to examine in animals the molecular events that occur in the course of the viral life cycle. We establish the feasibility of such a longitudinal analysis in vivo, document restriction in gene expression in alveolar macrophages and provide evidence that this restriction cannot be accounted for simply by gene dosage. The approach illustrated with visna should be of general applicability to other dynamic and molecular investigations of virus infection. PMID- 2983496 TI - Nucleotide sequence and structure of integrated bovine leukemia virus long terminal repeats. AB - Bovine leukemia virus (BLV) proviruses, harbored by the productively infected fetal lamb kidney (FLK-BLV) cell line, were cloned in bacteriophage lambda L47. The nucleotide sequence of the proviral long terminal repeats (LTR) with flanking cell and virus DNA have been determined. The BLV LTR is 531 bp in length and is bounded by the dinucleotides 5'-TG...CA-3', which are part of a 3-bp inverted repeat. The integrated provirus is flanked by 6-bp direct repeats of cellular DNA. A tRNApro primer binding site is present starting 2 bp downstream of the 5' LTR. In addition to sequencing integrated proviral DNA clones, the nucleotide sequence of a cDNA clone, representing the 3' end of genomic viral RNA, was determined; thus revealing the RNA polyadenylation site and R:U5 boundary within the LTR. Unlike most other retroviruses, a consensus polyadenylation signal, "AATAAA," is not located proximal to the BLV polyadenylation site. The RNA initiation site, defining the U3:R boundary, was located in the BLV LTR by S1 nuclease mapping. This site is approximately 25 bp downstream of an A + T-rich region which probably encompasses a Goldberg-Hogness ("TATAA") box and about 90 bp downstream of a potential "CCAAT" box. The BLV LTR possesses a U3 region of 204 bp, an unusually long R region of 241 bp, and a U5 region of 86 bp. PMID- 2983497 TI - Expression of an immediate early polypeptide and activation of a viral origin of DNA replication in cells containing a fragment of herpes simplex virus DNA. AB - A thymidine kinase cotransformation procedure has been used to introduce the sequences encoding the herpes simplex virus type 1 (HSV-1) immediate early protein, Vmw175, into permissive cells either in the presence or the absence of the adjacent origin of viral DNA replication. Cells transformed by either origin plus or origin-minus DNA were capable of expressing functional Vmw175 as indicated by their ability to complement the growth at the nonpermissive temperature of an HSV-1 mutant, ts K, containing a temperature-sensitive lesion in the Vmw175 gene. A proportion of the virus yield from cells transformed with the origin-plus, but not the origin-minus, plasmid exhibited a ts+ phenotype. The generation of ts+ virus correlated with an amplification of input plasmid DNA sequences which occurred following superinfection, suggesting that recombination between the ts mutant and the amplified viral DNA sequences had taken place. Encapsidation of the amplified DNA sequences was also detected, suggesting that in addition to a functional origin of replication and Vmw175 gene the transformed cells also retain the viral DNA packaging signals. PMID- 2983498 TI - In vivo and in vitro models of demyelinating disease. XI. Tropism and differentiation regulate the infectious process of coronaviruses in primary explants of the rat CNS. AB - The coronaviruses, ubiquitous in mammals, including man, manifest serotype related predeliction for different tissues. This presentation deals with specificity of the murine viscerotropic MHV3 and neurotropic JHMV for explanted cells from the CNS of newborn, inbred, Wistar-Furth rats. An unambiguous tropism of MHV3 for astrocytes and JHMV for oligodendrocytes is demonstrated. With the latter cell-virus interaction, relatively small differences in spatial density of oligodendrocytes influence profoundly the duration of persistence and virus yield. The in vitro temporal program of oligodendrocyte differentiation, monitored by induction of a myelin-related enzyme, 2':3'-cyclic nucleotide-3' phosphohydrolase, corresponds to that occurring in vivo (F. A. McMorris, J. Neurochem. 41, 506-515, 1983). It is complete within 15-21 days and is coincident with the onset of insusceptibility to disease caused by JHMV. Experimental elevation of intracellular cyclic-AMP levels, presumed to reflect oligodendrocyte differentiation, likewise suppresses JHMV replication without affecting that of MHV3 in astrocytes. On the basis of these data it is concluded that in vitro interaction of JHMV with oligodendrocytes reflects accurately the in vivo host control over the tropism and expression of this virus, thereby effecting the progressive, demyelinative disease, process. PMID- 2983499 TI - [Content of cyclic nucleotides, cholesterol and phospholipids in tumors of the large intestine]. AB - Tissues from tumor and normal intestine were examined in 44 patients suffering from colonic and rectal cancer. The study established elevated levels of cholesterol (chiefly, cholesterol esters) and phospholipids, as compared with normal tissue. Two groups were identified among the cancer patients on the basis of the ratio of K = (Formula: see text). Cases with a relatively low value of K revealed higher levels of body fat and hyperlipidemia. That was matched by relatively higher concentrations of cholesterol and its esters, but not phospholipids, in tumor tissue, as compared with normal one. The relationship between cyclic nucleotide metabolism disorders in neoplastic tissue and lipid metabolism in cancer patients is discussed. PMID- 2983500 TI - Acute hepatitis associated with Colorado tick fever. PMID- 2983501 TI - Cosyntropin stimulation in adrenal vein testing for aldosteronoma. PMID- 2983502 TI - [Therapy of multiple sclerosis]. AB - The routine therapy of multiple sclerosis (MS) in world-wide use today is comprised of four measures: Antiinflammatory and antiedematous treatment with ACTH or Synacthen, respectively, and corticosteroids: only during acute episodes. - High dosage, short duration, no long-term therapy. Immunosuppression with azathioprine (Imurek): Due to the relatively high risk only to use in malignant courses (frequent and severe bouts). Basic therapy with unsaturated fatty acids (sunflower oil, Naudicelle). Influencing circumscribed target symptoms (spasticity, micturition difficulties, constipation, etc.). In addition, physiotherapeutic, psychagogic and, if necessary, nursing and social measures are included. More than a decade's experience with ultrasound therapy of the lymphatic ring as developed by Selzer in over 300 MS-patients gives the impression of a reduction in bout frequency and severity. A statistical evaluation of therapeutic efficiency has so far been impossible for well-known disease-specific reasons, which hold true for all MS-treatment methods. Great practical importance within a foreseeable space of time may be reached by efforts to influence disturbance in nerve conduction and synaptic transmission as specifically caused by the demyelination process. The successful medicinal deceleration of sodium inactivation, inhibition of potassium activation and extension of the action potential, as well as specifically influencing the neurotransmitters responsible for the disturbed synaptic transmission could lead to a total recovery or improvement of dysfunction in a great many cases. Such a "global symptomatic therapy" might indeed not change the course of disease, but bring about great progress to the patient. PMID- 2983503 TI - Lithium and Wisconsin--a medicinal trip through history. PMID- 2983504 TI - [Free and conjugated plasma catecholamines, lactate behavior and oxygen uptake at rest and in staged physical exertion as well as alpha receptor density in intact thrombocytes in trained athletes]. AB - The influence of chronic static training on free and conjugated plasma catecholamines was investigated in 8 statically trained athletes (21 +/- 2 years of age) at rest and during incremental ergometric cycling. Plasma catecholamines are seen as biochemical indicators of the over-all sympathetic activity. Alpha-2 adrenoreceptors were additionally determined as one parameter of the organism's sensitivity to catecholamines. During modest and heavy exercise, free plasma noradrenaline and adrenaline are comparably low in statically and endurance trained athletes. They are lower than in untrained subjects. During exhaustion, however, free adrenaline responses of the statically trained athletes only amount to 30-50% of the concentrations observed in endurance trained athletes and untrained subjects. Free plasma dopamine and conjugated plasma catecholamines do not show any significant changes during ergometric exercise. Free and conjugated catecholamines correlate positively. Positive correlations are also observed between blood pressure and plasma catecholamines. These correlations however are clearer between conjugated catecholamines and blood pressure. This is also recognizable for the negative correlation between alpha-2-receptor density on intact platelets (1078 +/- 323 binding sites per cell) and conjugated catecholamines. In conclusion, statically trained athletes also show an alteration of sympathetic tone (catecholamines) comparable to that observed in endurance trained athletes. Maximal adrenaline responses however are lower in statically trained athletes than in endurance trained or untrained subjects. The alpha-adrenoreceptor density on intact thrombocytes seems to be increased in statically trained athletes. PMID- 2983505 TI - [Neurochemical mechanisms of the regulation of pain sensitivity]. PMID- 2983506 TI - [Physiological role of the adrenergic elements of the hypothalamus in regulating the hypothalamo-adenohypophyseal adaptation systems]. PMID- 2983507 TI - [Distribution and activity of nicotinamide-adenine dinucleotide nucleosidase in mammalian semen]. PMID- 2983508 TI - A study of bile secretion in conscious sheep. PMID- 2983509 TI - Laboratory experiments on oral vaccination of calves against rotavirus or coronavirus induced diarrhoea. PMID- 2983510 TI - Differential effects of epidural morphine in the treatment of cancer-related pain. AB - Fifty-five patients with pain associated with cancer were selected for long-term treatment with epidural morphine. Patients who had more than one type of pain within the same region were specially analysed concerning differential analgesic effects of the treatment, i.e. the patients served as their own control. Twenty eight of the 55 patients became pain free. In 21 patients alleviation of pain was complete only for one or two of several types of pain within the same area with a certain dose of epidural morphine. In six patients the treatment failed. An analysis revealed that the best response was obtained when the pain was continuous and originated from deep somatic structures. In co-existing continuous visceral pain or intermittent somatic pain originating e.g. from a pathological fracture, the outcome of the treatment was variable. Cutaneous pain, pain classified as neurogenic, and intermittent pain due to intestinal obstruction was only occasionally relieved. Ten of the patients had co-existing pain of non malignant origin and none of them was helped for that pain. The variable response to epidural morphine may indicate that different types of pain-producing stimuli engage different kinds of receptors which differ in affinity to morphine in the spinal fluid; it is also possible that some pain-mediating systems are non responsive to opiates. PMID- 2983511 TI - Cardiovascular studies during controlled baroreflex activation in the dog: II. Effects of metoprolol and enflurane. AB - An experimental model was designed to study circulatory drug effects with or without barostatic reflex influences. In dogs anaesthetized with chloralose, both carotid sinuses were perfused from a femoral by-pass either with systemic arterial pressure or with a pump in order to control the sinus pressure. Cardiac and aortic baroreceptors were denervated. I.v. metoprolol (240 micrograms X kg-1 + 102 micrograms X kg-1 X h-1) with constant carotid sinus pressure and thereby constant baroreceptor activation reduced cardiac output, heart rate, cardiac contractility and left ventricular stroke work index. Systemic vascular resistance increased. This response was independent of the degree of baroreflex activation. During the combined administration of enflurane (1.6% end-tidal concentration) and metoprolol, cardiac performance (cardiac output, stroke volume, heart rate, cardiac contractility and left ventricular stroke work index) was depressed independent of the degree of baroreflex activation. Barostatic reflexes, however, counteracted an observed decrease in systemic vascular resistance. This reflex vascular response was, during metoprolol-enflurane administration, associated with an increased left ventricular end-diastolic pressure. PMID- 2983512 TI - Experimental thalidomide neuropathy: the morphological correlate of reduced conduction velocity. AB - Morphological studies of experimental thalidomide neuropathy have thus far failed to show any significant structural changes. The present investigation was performed on sural nerves of female New Zealand white rabbits showing a reduction of sensory conduction velocity after oral treatment with thalidomide (100 mg/kg b. wt. per day) for a period of 33 weeks. Rabbits of the same strain and equal sex, weight, and number served as controls. Very few nerve fibers were undergoing Wallerian degeneration in both groups, experimental animals and controls. Morphometry, however, revealed a statistically significant reduction of the mean myelin thickness of sural nerve fibers in the thalidomide group of rabbits as compared to controls. The mean myelin thickness of the largest nerve fibers was also significantly smaller than in the control group. On the other hand, axonal diameters were not significantly altered. The association between the decrease of the sensory conduction velocity, the reduction of the myelin sheath thickness, and the chronic thalidomide application is discussed. PMID- 2983513 TI - Effect of long-term therapy with estrogen and progesterone on the endometrium of post-menopausal women. AB - We studied a group of 398 patients who had been receiving estrogen replacement therapy since 1976. Group I consisted of 138 patients who received Premarin (conjugated equine estrogen, Ayerst) in two different dosages (0.625; 1.25 mg) and progestational agents such as Neogest (Norgestrel, Schering), Duphaston (Dydrogesterone, Duphar) 5 mg and Primolut N (Norethisterone, Schering) 5 mg, for a period ranging between 7 and 21 days. Group II consisted of 106 patients who received Harmogen (Piperazine Oestrone Sulphate, Abbott) in two different dosages (1.5 mg; 2.25 mg) with the above-mentioned progestational agents for a period ranging between 7 and 21 days. Group III consisted of 154 patients who received Progynova (Oestradiol Valerate, Schering) in two different dosages (1 mg; 2 mg) with the above-mentioned progestational agents for a period ranging between 7 and 21 days. When increasing dosages of estrogen with progestational agents were added for 7 days, this produced cystic hyperplasia of the endometrium. When the duration of progestational agents was increased to 10 days or more, a more atrophic and secretory endometrium resulted and there was no incidence of cystic hyperplasia. PMID- 2983514 TI - The effect of lithium in vitro and in vivo on dopamine-sensitive adenylate cyclase activity in dopaminergic areas of the rat brain. AB - Lithium (5 and 20 mM) was found to inhibit the dopamine-stimulated cyclic AMP formation in homogenates from rat striatum and olfactory tubercle, leaving basal and fluoride-stimulated activities unaffected. The inhibition of dopamine stimulated adenylate cyclase was non-competitive and dose-dependent. However, in rats treated with lithium for four weeks, no alterations were found in basal, fluoride- and dopamine-stimulated adenylate cyclase activities. It is suggested that lithium interferes with hormonal stimulation of adenylate cyclase activity by an interaction with the process regulating the transfer of the receptor hormone stimulus to the adenylate cyclase enzyme. PMID- 2983515 TI - Diaphanography in breast carcinoma. Correlation with clinical examination, mammography, cytology and histology. AB - Histologically classified carcinoma was present in 110 breasts of 108 symptomatic women. The results of diaphanography (DPG) were correlated with those obtained by clinical examination (CE), mammography (M) and cytology (C). A tumour was palpable in 87 cases (79.1%). A false negative diagnosis was made in 17 cases (15.5%) using DPG, in 13 cases (11.8%) using M and in 15 cases (13.6%) using C, but in 12 of the latter cases (10.9%) the specimen was not representative. The validity of the findings using DPG and M was also analysed. The calculations were based on the results obtained from the present investigation and from a study of diaphanography in 163 cases of benign breast disorders. For DPG the sensitivity was 85 per cent, the specificity 91 per cent, the positive predictive value 86 per cent and the negative predictive value 90 per cent. For M the sensitivity was 86 per cent, the specificity 80 per cent, the positive predictive value 75 per cent and the negative predictive value 91 per cent. The specificity for diaphanography was significantly different from mammography (p less than 0.05). The use of both M and DPG reduced the number of false negatives from 11.8 per cent to 5.5 per cent. In conclusion, DPG has been demonstrated to be a useful adjunct to CE and M. PMID- 2983516 TI - Liver changes and complications in adult polycystic kidney disease. PMID- 2983517 TI - Synthesis of prostaglandins by vascular endothelial cells. PMID- 2983518 TI - Life-threatening acid-base disorders. PMID- 2983519 TI - Metrizamide CT myelography in cervical myelopathy and radiculopathy: correlation with conventional myelography and surgical findings. AB - Conventional myelography, metrizamide computed tomographic (CT) myelography, and surgical findings were correlated in 30 patients with cervical radiculopathy and/or myelopathy. In 60% of patients, metrizamide CT myelography provided significant additional information including better characterization of the abnormality, lateralization if the conventional myelogram was indeterminate, more definitive demonstration of cord atrophy, foraminal narrowing not appreciated on myelography, and demonstration of abnormalities distal to a myelographic block. In no case was a myelographic abnormality not detected on metrizamide CT myelography. In patients with cervical myelopathy, a cross-sectional diameter of the cord equaling less than 50% of the subarachnoid space is predictive of poor patient response to surgical intervention. PMID- 2983521 TI - Localizing peroneal nerve lesions. AB - The main result of a peroneal nerve lesion, foot drop, is so obvious that few patients can ignore its presence. However, the detailed knowledge of peroneal nerve anatomy needed to localize the lesion may tax even a seasoned clinician. Examination of the strength of five muscles--the tibialis anterior, peroneus longus and brevis, posterior tibial, gluteus medius and quadriceps--provides information necessary to localize the site of the neuropathy. This tentative localization can be confirmed by electromyography. Treatment is directed to the cause of the lesion and stabilization of the ankle if necessary. PMID- 2983520 TI - Treatment of pain with antidepressants. AB - Antidepressant drugs are now an important component of the analgesic pharmacopeia. Their ability to relieve pain is independent of the presence of clinical depression, and their efficacy has been documented in the treatment of migraine and tension headaches, neuropathic pain and chronic pain syndromes, including those associated with cancer. The tricyclic antidepressants are safe for use in patients with prolonged pain. The low doses required for analgesia are well tolerated and do not produce the side effects associated with the classic analgesic drugs. PMID- 2983522 TI - Improvement of hypocalcemic cardiomyopathy by correction of serum calcium level. PMID- 2983524 TI - Retinofugal projections of the big brown bat, Eptesicus fuscus and the neotropical fruit bat, Artibeus jamaicensis. AB - Retinal connections were studied in Eptesicus fuscus and Artibeus jamaicensis using anterograde axonal degeneration and autoradiographic techniques following unilateral enucleations and uniocular injections of radioactive amino acids. Although each retina projected bilaterally to the brainstem, the number of silver grains in the emulsion of autoradiographs indicated that nearly all fibers in the optic nerve entered the contralateral optic tract. Ipsilaterally, a major portion of the projection ended in the suprachiasmatic nucleus; caudal to the suprachiasmatic nucleus, the amount of label was so small that individual silver grains were counted to determine the location and quantity of label in other ipsilateral nuclei. In both species the retinal projection terminated bilaterally in the suprachiasmatic, dorsal lateral geniculate, ventral lateral geniculate, and pretectal olivary nuclei and contralaterally in the posterior pretectal nucleus, superficial gray layers of the superior colliculus, and nuclei of the accessory optic system. In Eptesicus the projection to the nucleus of the optic tract ended contralaterally, and in Artibeus it ended in this nucleus bilaterally. The results of this study revealed a basic theme in the optic projection of the two ecologically different microchiropterans. The results differed, however, in that the projection was larger and visually related nuclei were better developed in Artibeus. Such variations are presumed to relate to eye size and the relative use of vision by the two chiropterans. PMID- 2983523 TI - Mechanism of vasodilation by molsidomine. AB - Molsidomine is enzymatically metabolized in the liver to SIN-1 and readily converted into the active metabolite SIN-1A, which carries a free nitroso group. Evidence obtained in isolated circular strips from bovine coronary arteries indicates that SIN-1 increases cyclic guanosine monophosphate in close association with its relaxant effects in coronary strips under various pharmacologic conditions, suggesting that cyclic guanosine monophosphate mediates relaxation. Various nitrovasodilators act by the same mechanism, which is stimulation of guanylate cyclase. In this study the effect of nitroglycerin depended on the presence of a special thiol, cysteine, whereas SIN-1 was active also in the absence of cysteine. Cysteine deficiency was found to be associated with tolerance. After prolonged exposure to the drug, tolerance toward nitroglycerin developed in coronary strips that was antagonized by cysteine. SIN 1 produced no significant tolerance and was also fully active in nitroglycerin tolerant strips. We conclude that SIN-1 relaxes vascular smooth muscle by direct stimulation of guanylate cyclase, whereas nitroglycerin probably must be converted into a cyclase stimulator by a cysteine-dependent reaction. PMID- 2983526 TI - Detection of viral and chlamydial antigens in open-lung biopsy specimens. AB - The recovery of viruses and Chlamydia trachomatis from cell cultures and the detection of their antigens in impression smears prepared from open-lung biopsy (OLB) specimens from immunocompromised adults were compared. Touch impression smears were prepared on three slides, each containing eight wells. OLB tissue was homogenized (Stomacher) and inoculated into MRC-5, primary monkey kidney, and McCoy cell cultures. The direct and indirect immunofluorescence (IF) tests were used to detect antigens to the following organisms: herpes simplex virus, adenovirus, parainfluenza types 1 and 3, respiratory syncytial virus, cytomegalovirus (CMV), Chlamydia trachomatis, influenza types A and B, and varicella-zoster virus. Of 105 OLB specimens, 21 viral isolates (20%) were recovered in cell culture; 20 were CMV and one was an influenza virus type A (H3N2). Both culture and IF results were positive with 12 specimens, but in nine instances a virus was isolated and IF was negative or eight times culture results did not yield the organism but IF test results were positive. Chlamydia trachomatis was never isolated or detected by IF. The authors recommend that for optimal detection of CMV from OLB specimens a new rapid centrifugation-enhanced cell culture system be used in preference, or in conjunction with a preliminary IF screen of impression smears for CMV detection. PMID- 2983525 TI - Peripheral T-cell lymphoma: aggressive disease with heterogeneous immunotypes. AB - Eleven patients with mature or peripheral T-cell lymphoma (PTL), other than mycosis fungoides, were identified using an extensive battery of T- and B-cell markers. Eight cases had a histologic diagnosis of either diffuse large cell or mixed lymphoma, three of small cell type. All cases had one or more "mature" T antigens and an absence of B- and immature T-antigens. Assessment of T-antigens included E-rosettes (Er), anti-Leu 1-7 and Tdt. The authors delineated striking heterogeneity of T-antigen expression: 9 different immunotypes in 11 cases. Subset T-antigen assessment indicated T-helper neoplastic cells in five cases and T-suppressor in two. The remaining four had universal T-antigens alone. Seven cases appeared to have "novel" T-phenotypes not corresponding to normal T ontogeny phenotypes. Novel or idiosyncratic phenotypes may be a key characteristic of PTL. Since no single T-antigen, including Er and Er receptor (Leu-5), was expressed in all cases, a battery of monoclonal antibodies is necessary to detect PTL. Clinically, the authors found PTL unexpectedly aggressive, despite mature immunotype. Most patients were elderly (median age 69); all had extranodal disease with cutaneous involvement (six cases) most frequent. Responses to chemotherapy frequently proved transient, with median survival of nine months. A fulminant course was noted even with localized presentation. Clinical outcome suggests PTL requires new therapeutic strategies. PMID- 2983527 TI - Listeria monocytogenes sepsis and small cell carcinoma of the rectum: an unusual presentation of the acquired immunodeficiency syndrome. AB - A 26-year-old male homosexual initially presented with Listeria monocytogenes sepsis and a small cell carcinoma of the rectum. His subsequent course included esophageal candidiasis, Pneumocystis carinii pneumonia, and severe T-lymphocyte abnormalities on immunologic testing, consistent with the acquired immunodeficiency syndrome (AIDS). This represents the first case of AIDS associated with this unusual tumor and Listeria infection. PMID- 2983528 TI - Hypertension in children. Increased efficacy of technetium Tc 99m succimer in screening for renal disease. AB - Renal scintigraphy with technetium Tc 99m succimer (DMSA) and technetium Tc 99m pentetate (DTPA) was used to study 80 hypertensive pediatric and adolescent patients. Renal abnormalities such as asymmetry of function, size, or shape were identified in 13 patients. Both excretory urography and technetium Tc 99m pentetate studies were successful in detecting 54% of the abnormalities in patients studied; technetium Tc 99m succimer identified 92%. The accuracy of the latter was 96%, with a specificity of 97%. The ability of technetium Tc 99m succimer renal scintigraphy to identify accurately the presence or absence of renal abnormalities warrants its inclusion in the initial examination of pediatric and adolescent patients with hypertension. PMID- 2983529 TI - Fatal gastroenteritis associated with coronaviruslike particles. AB - The role of human enteric coronaviruses in infantile gastroenteritis is controversial. We detected coronaviruslike particles in the intestinal contents and within the epithelial cells of the ileum in a 15-month-old infant who had postmortem evidence of severe enteritis. Ultrastructural findings consistent with in vivo coronavirus replication in the human small intestine support a causative role for this agent in gastroenteritis. PMID- 2983530 TI - Hypothalamic-pituitary function in children with optic nerve hypoplasia. AB - We studied the clinical characteristics and hypothalamic-pituitary function in 23 patients with optic nerve hypoplasia (ONH), 6 months to 19 years old. All patients had decreased visual acuity and small optic discs; the septum pellucidum was absent in five of 19. Nine of 11 patients had minor EEG abnormalities, and two had microcephaly. The height, weight, and growth rate were normal in all patients 6 months to 3 1/2 years old. Hypopituitarism was found in 15 patients. Fasting and stimulated prolactin levels and the area under the prolactin curve after thyrotropin releasing hormone were significantly greater than in controls and in patients with idiopathic hypopituitarism. These results associate ONH with a high incidence of hypopituitarism, hyperprolactinemia, and neurologic abnormalities. The normal growth in the absence of measurable growth hormone suggests that hyperprolactinemia may stimulate growth temporarily and that a normal height in childhood or the presence of the septum pellucidum do not exclude the possibility of hypopituitarism. PMID- 2983531 TI - Reye's syndrome associated with adenovirus infections in infants. AB - The isolation of adenovirus from the liver of an infant with Reye's syndrome prompted a retrospective review to determine the role of this group of viruses in the cause of Reye's syndrome. A total of 34 patients with Reye's syndrome who were admitted to Vanderbilt University, Nashville, Tenn, during a ten-year period were identified. Six of the patients were younger than 1 year, and positive adenoviral cultures and increased serological responses were obtained in three of these children. The clinical and pathologic data from these three children suggest that adenovirus may be an important agent in initiating Reye's syndrome in the very young child. Further, the mitochondrial abnormalities present in the livers of the patients with adenovirus-associated disease tended to be mild in spite of severe clinical disease. PMID- 2983533 TI - Severe hyponatremia after colonoscopy preparation in a patient with the acquired immune deficiency syndrome. AB - Disseminated infections with cytomegalovirus have become an increasingly important cause of morbidity and mortality in patients with the acquired immune deficiency syndrome. Herein we describe a male homosexual with the acquired immune deficiency syndrome who became severely hyponatremic during bowel preparation for colonoscopy. Biochemical evidence of adrenal insufficiency and clinical evidence for cytomegalovirus colitis was found. We propose that the patient had clinically significant cytomegalovirus adrenalitis. A random urinary sodium concentration is suggested as a potential screening test before bowel preparation in such patients. PMID- 2983532 TI - Chronic T-cell lymphocytosis with neutropenia. Its association with Epstein-Barr virus infection. AB - Chronic T-cell lymphocytosis with neutropenia has primarily been reported in adults. We have cared for a 14-year-old patient with this condition for over five years. This patient consistently had greater than 90% lymphocytes in his differential cell count. The majority of these cells had the OKT8 phenotype. Serologic studies demonstrated evidence for a chronic Epstein-Barr infection with elevated antibody titers to both early antigen and to the viral capsid antigen. Patients presenting with this condition should be carefully evaluated for possible viral infection. PMID- 2983534 TI - Clear cell hepatocellular carcinoma: sudden death from severe hypoglycemia. AB - Hypoglycemia is a frequent paraneoplastic complication of hepatocellular carcinoma occurring in up to 30% of cases of this neoplasm. The clear cell carcinoma, a notable histological subtype of hepatocellular carcinoma, however, has been associated with hypoglycemia in only two previously reported patients from Japan. We report herein the first American case of clear cell hepatocellular carcinoma with severe hypoglycemia. The patient also suffered from hypercholesterolemia and erythrocytosis. Fine needle aspiration cytology, conventional histopathology, and immunohistochemical studies of this neoplasm are described. Possible explanations for the clear cell appearance and a comparison of the clinical and morphological features of clear cell and nonclear cell hepatocellular carcinoma is made. Potential pathophysiological mechanisms for the paraneoplastic complications of hypoglycemia, hypercholesterolemia, and erythrocytosis are discussed. PMID- 2983535 TI - Anomalous neutrophil granule distribution in a patient with lactoferrin deficiency: pertinence to the respiratory burst. AB - Neutrophils from a patient with lactoferrin deficiency were examined and the quantity and subcellular localization of protein markers were determined on Percoll density gradients. Distribution of azurophilic and specific granule markers was abnormal in that azurophilic granules were lighter than normal and appeared in the fraction of the gradient where normally the specific granules sediment. The specific granule membrane markers, cytochrome b-235 and its associated flavoprotein, were abnormally distributed in the gamma fraction, the site of the plasma membrane marker alkaline phosphatase. Thus, the b-cytochrome flavoprotein complex had either been incorporated into the plasma membrane or was still present in the membranes of granules that were abnormally light and cosedimented with the plasma membranes. This is of particular interest in regard to the patient's respiratory burst oxidase function, since the b cytochrome/flavoprotein complex normally translocates from the specific granules to the plasma membrane to constitute the active respiratory burst oxidase. The functional consequences of this abnormal distribution are discussed, as is the importance of characterizing both intragranular enzymatic markers and granule membrane proteins to define granular disorders. PMID- 2983536 TI - Gene mapping of Malaysian alpha thalassemias with alpha and zeta globin gene probes. AB - Restriction enzyme analysis of the alpha and zeta globin genes was carried out in four cases of Hb Bart's hydrops fetalis, in three patients with Hb H disease without Hb CoSp, in three patients with Hb H disease with Hb CoSp, in 47 individuals with alpha thalassemia trait, and in 47 normal individuals. All four cases of Hb Bart's hydrops fetalis resulted from deletions of alpha 1 and alpha 2 globin genes which did not extend to the psi zeta 1 and zeta 2 globin genes. The same type of deletion was observed in alpha thal1 carriers, but two newborns (one Malay and one of Chinese extraction) had a nondeletion type of alpha thal1 which was confirmed by quantitative alpha globin gene analysis. In addition, two other newborns diagnosed as alpha thal1 trait carriers (one Malay, one Chinese) were shown to have a deletion of both alpha globin genes by quantitative alpha globin gene analysis, but further testing with zeta globin gene probe failed to reveal an abnormal fragment length characteristic of an alpha globin gene deletion. We believe that this last condition is due to a large deletion which includes all alpha globin genes and all zeta globin genes on the same chromosome. On another front, Bgl II restriction analysis of all four Hb Bart's hydrops fetalis cases and the alpha thal1 trait carriers showed a 10.5-kb Bgl II restriction fragment, in the hydrops fetalis as a single band, while in the carriers this 10.5-kb fragment was accompanied by the usual normal 12.5-kb and 11.3-kb fragments. We report that this 10.5-kb fragment, previously thought to be specific for the Southeast Asian alpha thal1 gene deletion, is also common in normal individuals. Nevertheless, digestion with other enzymes can clearly differentiate the alpha thal1 and normal genotypes. We distinguish the findings in the alpha thalassemias from the extensive DNA polymorphism in the region of the alpha and zeta globin genes. PMID- 2983537 TI - Multiple myeloma with crystalline inclusions in most hemopoietic cells. AB - Intracytoplasmic inclusions were observed at light and electron microscopy in the cells of most of the hemopoietic series, including erythroblasts, in a case of multiple myeloma IgG kappa. The inclusions were of crystalline nature, surrounded by smooth membrane. An indirect immunoperoxidase method on thick sections confirmed their immunoglobulin G kappa nature. The fact that crystals were observed in various hemopoietic series suggests that, except for the plasma cells, their presence is more likely due to phagocytosis than to synthesis and, therefore, that they are not a clone marker. PMID- 2983538 TI - Acute nonlymphocytic leukemia following treatment of testicular cancer and gastric cancer with combination chemotherapy not including alkylating agents: report of two cases. AB - Acute nonlymphocytic leukemia following combination chemotherapy not including alkylating agents or radiotherapy was observed in one patient treated for testicular cancer and in another treated for gastric cancer. Both patients presented clinical, cytologic, and cytogenetic findings uncharacteristic for secondary acute nonlymphocytic leukemia. It is discussed whether these two cases of leukemia indicate a risk of secondary leukemia following chemotherapy with cisplatinum and adriamycin. PMID- 2983539 TI - Alpha globin gene rearrangements in Polynesians are not associated with malaria. AB - Using gene mapping, 16.3% of Polynesians were shown to have alpha thalassemia. These results are surprising since malaria is not found in Polynesia. Moreover, triplicated alpha gene rearrangements were identified in a further 7.7%, a frequency not seen in other populations. PMID- 2983540 TI - Sodium potassium ATPase activity in human rectal mucosa with and without renal insufficiency. AB - Studies in rats have shown that fecal potassium excretion and colonic mucosa Na-K ATPase activity are elevated during dietary potassium loading and in chronic renal insufficiency. We studied Na-K-ATPase activity in human rectal mucosa in normal subjects as well as in patients with chronic renal insufficiency (creatinine clearance 2 to 72 mL/min). In normals, Na-K-ATPase activity was 4.34 +/- 0.83 mumol P/mg protein. After 2 weeks on a potassium intake of 300 mmol/d the mean activity did not differ significantly from the control value (2.49 +/- 1.30). In none of the patients with renal failure was Na-K-ATPase activity beyond the range found in the normal subjects, irrespective of serum potassium; the mean activity was 3.50 +/- 0.85. Like others, however, we found a two-fold increase in Na-K-ATPase activity in potassium loaded rats. Possible explanations for these differences are discussed. PMID- 2983541 TI - HLA-B18 is associated with decreased levels of isoproterenol-stimulated cAMP in lymphocytes. AB - One hundred fifty-nine individuals were typed for HLA-A and B antigens and levels of isoproterenol-stimulated, lymphocyte cAMP. No significant age, sex, or caffeine effects on the natural log of the lymphocyte cAMP variable (ln cAMP) were found. A comparison of mean ln cAMP levels between individuals who carried a particular antigen (homozygous or heterozygous) and individuals who did not carry the antigen identified a highly significant decrease in ln cAMP levels associated with the HLA-B18 antigen. We estimated that 18.9% of the variability in ln cAMP was attributable to the HLA-B18 antigen. In addition, 38% of the variability in ln cAMP was attributable to factors that aggregate in families that were independent of the HLA-B18 effect. A weaker association of A10 with lymphocyte cAMP might be due to linkage disequilibrium between A10 and B18. PMID- 2983542 TI - Hereditary thrombosis in a Utah kindred is caused by a dysfunctional antithrombin III gene. AB - Maximum likelihood analysis of linkage between antithrombin III (ATIII) DNA polymorphisms and ATIII deficiency in a large Utah kindred suggests that thrombotic disease in this family is caused by a dysfunctional ATIII gene. ATIII deficient family members were identified on the basis of: (1) reduced anticoagulant activity and (2) the presence of an electrophoretically abnormal inhibitor molecule in their plasmas. Affected individuals have two copies of the ATIII structural gene, and both alleles appear normal at the resolution of whole genome Southern blotting. However, family studies revealed statistically significant cosegregation of ATIII-deficiency trait and a particular ATIII DNA polymorphism haplotype (lod = 3.35; theta = 0.0); this result is consistent with the presence of a dysfunctional ATIII gene on a chromosome of the +, S haplotype. PMID- 2983543 TI - Discrimination among the human beta A, beta S, and beta C-globin genes using allele-specific oligonucleotide hybridization probes. AB - Synthetic nonadecanucleotides complementary to the human beta A-, beta S-, or beta C-globin sequences were used as hybridization probes to screen human genomic DNA samples for these genes. The oligonucleotides were 32P-labeled and used as probes to genotype restriction endonuclease digests of human genomic DNA. The data obtained show that hybridization with oligonucleotide probes, unlike restriction fragment length polymorphism (RFLP) analysis or direct restriction enzyme digestion, can be used to directly distinguish among the three alleles of beta-globin, beta A, beta S, and beta C, when present either in one (heterozygous) or two copies. PMID- 2983544 TI - A discordant sibship analysis between beta-NGF and neurofibromatosis. AB - A new restriction fragment length polymorphism 5' to the beta-nerve growth factor (beta-NGF) gene has been found in proximity to the BglII polymorphism, and both polymorphisms are detectable with an EcoRI 7-kilobase (kb) subclone. Absence of the TaqI recognition site lengthens the 4.3-kb and 1.7-kb hybridizing fragments to 6 kb, and the alleles are in Hardy-Weinberg equilibrium with frequencies of 83% and 17%, respectively. Previous research has suggested that NGF is involved in disseminated neurofibromatosis (NF). We found four informative disseminated NF families with the two beta-NGF polymorphisms and have provided clearcut evidence against beta-NGF gene alteration in these families. If disseminated NF is found to be heterogeneous at a molecular level, more families should be tested to further rule out any role for beta-NGF in this syndrome. PMID- 2983545 TI - The use of multiple restriction fragment length polymorphisms in prenatal risk estimation. I. X-linked diseases. AB - An analytical procedure for estimating the risk of X-linked diseases based on presence/absence of a series of restriction sites is presented. Multiple-locus linkage phase of the carrier mother is first inferred from previous offspring, from parents, and by molecular means. Bayesian risk estimates are then obtained using this information and the recombination-segregation distribution. The improvement afforded by using multiple flanking markers rather than a single marker is dramatic. Whereas the upper bound on the probability that a family will be informative using a single diallelic X-linked marker is .5, in the case of m markers, the bound on the probability of an informative family becomes 1 - .5m. With a single linked marker, the precision in the risk estimate is bounded by the frequency of recombination, whereas the requirement of very tight linkage is relaxed somewhat when multiple flanking markers are used. Recombination interference and multiple-locus linkage disequilibria can further improve the risk estimates, but it is important to understand how the statistical confidence in these parameters affects the reliability of the risk estimates. PMID- 2983546 TI - Stability of an injectable disulfiram formulation sterilized by gamma irradiation. AB - Stability of an injectable disulfiram suspension sterilized by gamma(gamma) irradiation was tested. Single doses of disulfiram powder in plastic syringes were subjected to 50,000 rads of gamma radiation. Culture media were inoculated with the irradiated drug to test for growth of bacteria, fungi, and mycobacteria. The irradiated drug and nonirradiated controls were analyzed by high-performance liquid chromatography (HPLC) for disulfiram and its major degradation product, diethyldithiocarbamate (DDC). Ultraviolet absorption spectra of irradiated and nonirradiated disulfiram were obtained. No organisms grew in any of the culture media. HPLC analysis indicated that disulfiram content of the irradiated specimens was not reduced, and DDC was not detected. There were no important differences between the ultraviolet spectra of the irradiated and nonirradiated samples. Disulfiram can be sterilized by gamma irradiation without chemical degradation. PMID- 2983547 TI - Role of etoposide-based chemotherapy in the treatment of patients with refractory or relapsing germ cell tumors. AB - Forty-nine patients with metastatic germ cell tumors were treated with etoposide 100 mg/m2 and cisplatin 20 mg/m2 intravenously each day for five days as "salvage" chemotherapy. Forty-seven patients had received standard induction regimens for metastatic germ cell tumors before receiving etoposide and cisplatin. Four patients were treated after surgical resection of a single site of relapse (Group I). Forty-five patients had measurable or evaluable disease at the time of treatment. In 17 patients with evaluable disease who had either achieved a prior complete remission or received no prior cisplatin (Group II), eight (47 percent) complete and four (24 percent) partial remission were observed. In 28 patients who had never achieved a prior complete remission (Group III), no complete and five (18 percent) partial responses were observed. Seven of 21 patients in Groups I and II and none of 28 patients in Group III remain alive and free of disease. Assuming prior treatment with cisplatin-based chemotherapy, these data and a review of the published experience with similar salvage regimens for patients with relapsing or refractory germ cell tumors suggest that combination chemotherapy based on etoposide and cisplatin is effective primarily in those patients who achieved a prior complete remission. Such therapy is ineffective in the absence of a prior complete remission probably because the patients have tumors that are largely resistant to cisplatin. Observed responses are probably due to etoposide alone. Investigational therapies should be pursued in those patients whose disease is refractory to current induction regimens. PMID- 2983548 TI - Spectrum of pulmonary diseases associated with the acquired immune deficiency syndrome. AB - Over a four-year period, 130 patients with the acquired immune deficiency syndrome were studied to assess the incidence and spectrum of pulmonary disease associated with this illness. In 61 patients (47 percent), respiratory abnormalities were either present on admission or later developed. Multiple pathologic processes were present simultaneously in 24 patients and serial pulmonary problems developed in seven patients. Infection was the most common cause of pulmonary parenchymal disease and was due to Pneumocystis carinii (35 patients), cytomegalovirus (21 patients), Mycobacterium avium-intracellulare (13 patients), and bacteria (four patients). Noninfectious causes of parenchymal lung diseases were also frequently seen and included Kaposi's sarcoma (eight patients), non-specific pneumonitis (seven patients), and adult respiratory distress syndrome (four patients). Significant pleural disease was present in six cases and was usually related to Kaposi's sarcoma. A bronchospastic disorder developed in four patients. Pulmonary function tests, in particular the diffusing capacity and the difference between rest and exercise alveolar-arterial oxygen tension, were helpful in screening for pulmonary diseases. Patterns of clinical features and radiographic abnormalities were recognized and suggested specific diagnoses. Overall mortality from respiratory causes identified during the study was 41 percent, but varied markedly with the etiologic agent. Respiratory failure, however, carried a 100 percent mortality despite the underlying cause. PMID- 2983549 TI - Fever, abdominal pain, and hepatomegaly in a 66-year-old man. PMID- 2983550 TI - Electron microscopic demonstration of lysosomal inclusion bodies in lung, liver, lymph nodes, and blood leukocytes of patients with amiodarone pulmonary toxicity. AB - The mechanism of amiodarone-induced pulmonary toxicity is unknown. Two cases of amiodarone pulmonary toxicity are presented in which abnormal inclusion bodies containing whorls of membrane were seen on electron microscopy of extrapulmonary tissues. These cytoplasmic lysosomal inclusion bodies were observed in lymphocytes, plasma cells, granulocytes, tissue macrophages, and hepatocytes. These widespread histopathologic changes in extrapulmonary tissues and in a variety of cell types are similar to more extensively investigated findings in animal models that are thought to represent a drug-induced lysosomal storage disease, phospholipidosis. PMID- 2983551 TI - Electrocardiographic finding simulating acute myocardial infarction in a compound metabolic aberration. AB - A patient with hypokalemic metabolic alkalosis, hypophosphatemia, and hypomagnesemia/hypocalcemia is described. Electrocardiography demonstrated the pattern of acute anterior myocardial infarction. Further evaluation revealed that the patient had not actually had the acute myocardial infarction and that the electrocardiographic change was a mere simulation. The possible role of hypomagnesemia in the pathogenesis of the electrocardiographic change and the interrelation between the metabolic disturbances noted are discussed. PMID- 2983552 TI - Regulation versus modulation in GnRH receptor function. AB - Serum luteinizing hormone (LH) concentration after exposure to gonadotropin releasing hormone (GnRH) indicates that an instantaneous increase occurs in the rate of release of LH directly from the anterior pituitary, as measured dynamically during superfusion in vitro. On the other hand, estradiol-17 beta (E2) alone shows no such instantaneous effect on LH release rate (at least for the first four hours), in either physiologic or pharmacologic concentrations. At the same time, brief (ten to 30 minute) exposure of isolated anterior pituitary plasma membranes to physiologic concentrations of E2 significantly alters the binding of a fully biologically active 125I-GnRH to its plasma membrane receptor protein. In order to characterize the effect of E2 on GnRH binding further, we preincubated dispersed bovine anterior pituitary cells for six hours in the presence or absence of physiologic concentrations of E2 (10(-10)M). Following preincubation in the presence of E2, the cell suspension was incubated for 30 minutes with physiologic concentrations (5 X 10(-11) - 5 X 10(-10)M) of a fully biologically active 125I-GnRH. The treatment, at least, doubled the number of biologically important high affinity GnRH binding sites (Kd's = 7.5 X -10(-11) - 4.5 X 10(-10)M), and changed the binding capacity of some of the binding sites up to three fold, which altered the cooperativity of GnRH-receptor interaction. Thus, the interaction of E2 with GnRH at the level of GnRH receptor is mandatory for the short-term pituitary effect of E2 on LH release in vitro and in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983553 TI - Estrogen increases beta-adrenergic binding in the preterm fetal rabbit lung. AB - Tritium-labeled dihydroalprenolol was used to quantify beta-adrenergic-receptor sites in day 28 fetal rabbit lung tissue. Each of the amniotic sacs of pregnant New Zealand White rabbits on day 26 of gestation was injected in vivo with estrogen (estradiol phosphate, 1.6 micrograms) in one horn and normal saline solution in the contralateral one. The animals were put to death 48 hours later and the fetal lung tissues were assayed. Estrogen increased the number of beta adrenergic-receptor sites in the treatment group compared to the control group (216 versus 163 fmol/mg of protein, p less than 0.02 by paired t test). In the presence of estrogen, beta-adrenergic-receptor activity is enhanced in the preterm fetal rabbit. This effect may be implicated in the beta-adrenergic mediation of phospholipid synthesis and release in fetal alveolar cells. PMID- 2983554 TI - Estradiol potentiation of gonadotropin-releasing hormone responsiveness in the anterior pituitary is mediated by an increase in gonadotropin-releasing hormone receptors. AB - In order to investigate the mechanism by which 17 beta-estradiol potentiates the action of gonadotropin-releasing hormone on the anterior pituitary in vitro, cultured pituitary cells from immature female rats were used as the model system. Cultures exposed to estradiol at concentrations ranging from 10(-10) to 10(-6) mol/L exhibited a significant augmentation of luteinizing hormone release in response to a 4-hour gonadotropin-releasing hormone (10 mumol/L) challenge at a dose of 10(-9) mol/L compared to that of control cultures. The estradiol augmentation of luteinizing hormone release was also dependent on the duration of estradiol exposure. When these cultures were incubated with tritium-labeled L leucine, an increase in incorporation of radiolabeled amino acid into total proteins greater than that in controls was observed. A parallel stimulatory effect of estradiol on iodine 125-labeled D-Ala6 gonadotropin-releasing hormone binding was observed. Cultures incubated with estradiol at different concentrations and various lengths of time showed a significant increase in gonadotropin-releasing hormone binding capacity and this increase was abrogated by cycloheximide. Analysis of the binding data showed that the increase in gonadotropin-releasing hormone binding activity was due to a change in the number of gonadotropin-releasing hormone binding sites rather than a change in the affinity. These results suggest that (1) estradiol treatment increases the number of pituitary receptors for gonadotropin-releasing hormone, (2) the augmentary effect of estradiol on luteinizing hormone release at the pituitary level might be mediated, at least in part, by the increase in the number of binding sites of gonadotropin-releasing hormone, and (3) new protein synthesis may be involved in estradiol-mediated gonadotropin-releasing hormone receptor induction. PMID- 2983556 TI - Prevalence of papillomarvirus infection in colposcopically directed cervical biopsy specimens in 1972 and 1982. AB - A total of 1264 consecutive cervical biopsy specimens obtained at the Parkland Memorial Hospital Dysplasia Clinic during 1972 were reviewed. Histopathologic specimens were assessed with special reference to changes induced by human papillomavirus. In 1972, only 0.7% of biopsy specimens were reported as consistent with human papillomavirus infection. Upon review, however, 36.5% of these specimens were found to demonstrate histologic criteria for the diagnosis of human papillomavirus infection. Approximately half of biopsy specimens reclassified as human papillomavirus were originally interpreted as inflammation; the others were interpreted as cervical intraepithelial neoplasia. Patients with human papillomavirus infection were significantly younger than patients with cervical intraepithelial neoplasia (24.9 versus 30.2 years). These findings were compared with 965 cervical biopsy specimens obtained in 1982. Thirty-four percent of these biopsy specimens revealed human papillomavirus infection. These observations support the concept that human papillomavirus infection of the cervix is not a new entity but a previously unrecognized finding whose prevalence has been relatively stable over a 10-year period. PMID- 2983555 TI - Modulation of thymosin beta 4 by estrogen. AB - The endocrine thymus produces several hormone-like peptides (generically termed thymosins) which control development of the thymic-dependent lymphoid system and participate in the process of immune regulation. In addition, recent literature supports the hypothesis that gonadal steroids in general and estrogens in particular affect the immune system. To determine whether steroid hormones modulate secretion of thymic peptides, basal concentrations of thymosins alpha 1 and beta 4 were determined by radioimmunoassay in morning blood samples from 87 women in various clinical states. Basal concentrations of thymosin alpha 1 were similar in all women sampled. Basal levels of thymosin beta 4 were similar in normal women during the early follicular phase, women with premature ovarian failure, postmenopausal women not receiving estrogen, and individuals with gonadal dysgenesis. However, the marked variability of basal levels in premature ovarian failure and in postmenopausal women suggests that these groups are quite heterogeneous. Thymosin beta 4 concentrations were reduced in castrated women not receiving estrogen and were decreased more in both postmenopausal women and castrated women who were on chronic estrogen therapy. These data suggest that estrogens can modulate the circulating levels of thymosin beta 4 but not of thymosin alpha 1. We do not yet know whether sex steroids modulate secretion of other thymic peptides. PMID- 2983558 TI - Comparison of survival rates for patients with uveal melanoma after treatment with proton beam irradiation or enucleation. AB - We compared survival experiences for three groups with uveal melanoma: (1) 120 patients treated by proton beam irradiation from 1975 to 1981; (2) 235 patients treated by enucleation from 1953 to 1973 (enucleation group 1); and (3) 161 patients treated by enucleation from 1975 to 1981 (enucleation group 2). The following variables were predictive of melanoma-related deaths. TREATMENT: rate ratio (ratio of the rate of death for one category of the variable relative to the rate of death for another category) of 6.32 (95% confidence interval 1.70 to 23.51) for enucleation group 1 vs proton beam, 3.06 (0.81 to 11.54) for enucleation group 2 vs proton beam, and 2.07 (1.05 to 4.07) for enucleation group 1 vs enucleation group 2. Size: rate ratio of 3.61 (1.51 to 8.64) for large (16+ mm) vs small (less than 11 mm) tumors. LOCATION: rate ratio of 3.19 (1.56 to 6.53) for tumors anterior vs posterior to the equator. When death from all causes was evaluated as the outcome, the estimated rate ratios were similar. When melanoma metastasis was evaluated as the outcome, the estimated rate ratios were reduced in magnitude. PMID- 2983557 TI - A prospective study of herpes simplex virus infection in a defined population in Houston, Texas. AB - A seroepidemiologic study was conducted to determine the incidence of antibodies to herpes simplex virus in a population of middle-class Caucasian women. In utero exposure to diethylstilbestrol did not influence the frequency of herpes simplex virus infection. The frequency of infection increased with age and number of sex partners and a 50% increase in incidence of herpes simplex virus type 2 infection was noted during the 4-year period of the study. Subclinical herpes simplex virus type 2 infections apparently occurred in some women without preexisting antibodies to herpes simplex virus type 1. PMID- 2983559 TI - Unusual inclusions in stromal macrophages in a case of gelatinous drop-like corneal dystrophy. AB - A 35-year-old man developed gelatinous drop-like corneal dystrophy at age 20. Amyloid deposits were located mainly in the anterior two thirds of the cornea. Ultrastructurally, amyloid fibrils were demonstrated beneath the corneal epithelium and in the cytoplasmic inclusions of the stromal macrophages. PMID- 2983561 TI - Ceramics in tympanoplasty. PMID- 2983560 TI - Clinicopathologic responses in cats with feline leukemia virus-associated leukemia-lymphoma treated with staphylococcal protein A. AB - Purified protein A from Staphylococcus aureus Cowan I was injected intraperitoneally or was incorporated in filters ex vivo through which plasma from cats with feline leukemia virus (FeLV)-associated leukemia-lymphoma was passed. Before treatment, 65% of the FeLV-infected cats were anemic, and 70% were thrombocytopenic. Concomitant infections, or immune-mediated disease, was common. During treatment 50% of the cats with FeLV-associated disease improved objectively with normal posttreatment hematocrits, thrombocyte and leukocyte counts, disappearance of dysplastic hematologic elements, and correction of marrow dyscrasias. A 33% response to treatment occurred in cats with unequivocal manifestations of malignant disease and was characterized by reductions in tumor size and marrow and peripheral blood neoplastic cell populations. Clearance of FeLV viremia was documented in 28% of the treated cats. The several possible mechanisms by which treatment with staphylococcal protein A causes reduction in the extent of malignant disease are considered. PMID- 2983562 TI - Experimental studies on pathology and therapy of otospongiosis. PMID- 2983564 TI - Comparison of subunits of cardiac, brain, and kidney Na+-K+-ATPase. AB - Na+-K+-ATPase is in low abundance in cardiac tissue. Therefore, we utilized antibodies to detect the cardiac Na+-K+-ATPase subunits and to compare their characteristics with those of kidney and brain Na+-K+-ATPase subunits. By using crude preparations of heart membranes as well as purified sarcolemmal membranes from guinea pig hearts, we resolved peptides by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, blotted them onto diazotized paper, and detected Na+-K+-ATPase subunits with antibodies generated against highly purified kidney Na+-K+-ATPase holoenzyme. We tested the hypothesis that the two families of ouabain-binding affinities described in heart are due to two forms of alpha-subunit, analogous to the two forms with different affinities for ouabain described in brain. Although the antibodies did detect two forms of catalytic subunit in brain (alpha and alpha +), only one form of alpha was detected in the heart membranes, with the same electrophoretic mobility as kidney alpha. Cardiac beta-subunit could also be detected with the antikidney antibodies. It had a similar electrophoretic mobility to that described for kidney beta, whereas brain beta had a higher mobility. PMID- 2983563 TI - Calcium-dependent secretory and redox response to CCK-8 in isolated perfused rat pancreas. AB - Continuous stimulation with 8 pM cholecystokinin octapeptide (CCK-8) induced a gradual increase in pancreatic protein output and little if any change in redox state of cytochromes aa3, b, and c + c1. The protein output was completely abolished when CaCl2 was removed from the perfusing and bathing solution. Continuous stimulation with 200 pM CCK-8 induced the rapid and largest protein output and a distinct reduction of cytochromes and nicotinamide nucleotides. These responses were partially decreased in the Ca2+-deficient environment and enhanced immediately after the replacement with the standard solution. These and other results are compatible with the view that reduction of electron transfer system in the pancreatic acinar cell may be induced by stimulation with the secretagogue in pharmacological concentration and that the reduction may coincide with uptake and retention of cytoplasmic excess Ca2+ by mitochondria. PMID- 2983565 TI - Serotonin, histamine, and norepinephrine mediation of endothelial and vascular smooth muscle cell movement. AB - The effects of serotonin (5-HT), histamine (H), and norepinephrine (NE) on bovine aortic endothelial cell (BAE) and vascular smooth muscle cell (VSM) movement were quantitated using a phagokinetic tracking assay. BAE movement was significantly reduced by 5-HT (31%), H (48%), and NE (62%) at concentrations ranging from 10( 10) to 10(-4) M (P less than 0.01). VSM motility was significantly enhanced by 5 HT (17%) and H (25%) at concentrations of 10(-8) and 10(-4)M, respectively (P less than 0.01). NE (10(-4)M) reduced VSM motility by 63% (P less than 0.01). Ketanserin (1 microM) reduced the 5-HT-associated inhibition of BAE movement by 75% (P less than 0.01). Diphenhydramine (1 microM) reduced the H-associated inhibition of BAE movement by 66% (P less than 0.01). Propranolol (1 microM) reduced NE-associated inhibition of BAE movement by 50% (P less than 0.01) and 5 HT-associated inhibition by 45% (P less than 0.05), but phenoxybenzamine had no significant effect. The results suggest 1) BAE and VSM are affected qualitatively in opposite ways by 5-HT and H; 2) the H-associated inhibition of BAE movement appears to be at least partially mediated by H1 receptors; 3) the 5-HT-associated inhibition of BAE movement may be mediated directly by a 5-HT binding component but indirectly by beta-adrenergic receptors; and 4) the NE-associated inhibition of BAE movement appears to be mediated by beta-adrenergic receptors. PMID- 2983566 TI - Effect of vitamin D3 metabolites on calcium and phosphorus metabolism in chick embryos. AB - The biochemical nature of the physiological defect found in chick embryos from hens supported on 1,25-dihydroxyvitamin D3 as their sole source of vitamin D is described. Vitamin D-deficient hens (44-wk-old) were divided into six groups of five and dosed daily for 19 wk with either 2.0 micrograms of 25-hydroxyvitamin D3, 2.0 micrograms of 24,24-difluoro-25-hydroxy-vitamin D3, 0.4 micrograms of 1,25-dihydroxyvitamin D3, 2.0 micrograms of 24,25-dihydroxyvitamin D3, 0.4 micrograms of 1,25-dihydroxyvitamin D3 plus 2.0 micrograms of 24,25 dihydroxyvitamin D3, or vehicle only. Normal embryonic development was found in eggs from hens given 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3, whereas embryos from hens given 1,25-dihydroxyvitamin D3, 24,25-dihydroxyvitamin D3, or their combination were abnormal and failed to hatch. Embryos from hens fed 1,25-dihydroxyvitamin D3 and/or 24,25-dihydroxyvitamin D3 had vitamin D deficiency: low bone ash, low plasma calcium, low total body calcium, and extremely high plasma phosphorus. Because the shell is the major source of calcium for the developing embryo, calcium transport from the shell to the embryos across the chorioallantoic membrane apparently fails, giving rise to the observed defects in embryonic development. PMID- 2983568 TI - Jejunal-ileal differences in dopaminergic but not alpha-adrenergic antisecretory effects. AB - Clonidine, an alpha-adrenergic agonist, and apomorphine, a dopaminergic agonist, inhibit water secretion in rat intestine induced by dibutyryl cAMP (DBcAMP) in situ. Apomorphine reversed DBcAMP-induced secretion in ileum and jejunum with ED50 values of 90 and 200 nM, respectively. ED50 values for clonidine's antisecretory effect were 32 and 2 microM in ileum and jejunum, respectively. The antisecretory effects of both clonidine and apomorphine were blocked by the alpha adrenergic antagonist phentolamine and by the dopaminergic antagonists halo peridol and sulpiride. Phentolamine was about 10-fold more potent against clonidine than against apomorphine in the ileum and the jejunum. Haloperidol and sulpiride were at least 10-fold more potent against apomorphine than against clonidine in the ileum. In contrast, in the jejunum, haloperidol was equally potent in blocking effects of both agonists, while sulpiride did not block the apomorphine effect. The data are consistent with the presence of both alpha 2 adrenergic and DA2-dopaminergic receptors that stimulate absorption in the ileum. In the jejunum, antisecretory effect appears to be mediated by an action on either an alpha 2-adrenergic or a dopaminergic receptor with characteristics different from those of peripheral DA1- or DA2-receptors. PMID- 2983567 TI - Influence of renal perfusion pressure on alpha- and beta-adrenergic stimulation of renin release. AB - Experiments were performed in pentobarbital-anesthetized dogs to 1) determine if neural stimulation of renin release can be mediated by renal alpha-adrenoceptors at renal nerve stimulation (RNS) frequencies that have little or no effect on total renal blood flow (less than or equal to 1.2 Hz) and 2) ascertain whether alpha-adrenergic control of renin release is affected by renal perfusion pressure (RPP). The renal nerves were electrically stimulated both in the absence of RPP control and with RPP controlled near 85 mmHg. Decreased RPP lowered the threshold for neurogenic stimulation of renin release from less than or equal to 1.2 to 0.3 Hz. beta-Adrenoceptor blockade with propranolol blunted the renin secretion rate (RSR) response to graded RNS (0.3-5.0 Hz), but the extent of inhibition during low-frequency RNS was dependent on RPP. Propranolol prevented increased RSR at 0.6-1.2 Hz RNS when RPP was 111-120 mmHg but not when RPP was 85 mmHg. Combined alpha- and beta-blockade with prazosin and propranolol totally prevented increased RSR during 0.6-1.2 Hz RNS at reduced RPP. In summary, both alpha- and beta-adrenoceptors mediate neural stimulation of renin release at RNS frequencies that do not decrease total renal blood flow when RPP is 85 mmHg. PMID- 2983569 TI - Maintenance of renal autoregulation during infusion of aminophylline or adenosine. AB - Adenosine has been postulated to link control of glomerular filtration rate (GFR) and renal blood flow (RBF) with changes in renal metabolism. In the present study, we examined the role of adenosine in renal autoregulation by comparing the responses of normal anesthetized dogs to step decreases in renal artery pressure (RAP) to the response obtained after receptor blockade of adenosine with aminophylline or by flooding the kidney with exogenous adenosine. In six dogs at normal RAP, intrarenal infusion of aminophylline (10 mumol/min) did not alter renal hemodynamics. GFR and RBF were well autoregulated (greater than 90% of control) at RAP values equal to or greater than 85 mmHg before and after aminophylline. At RAP equal to 75 mmHg, GFR and RBF decreased by 27 +/- 10 and 20 +/- 8%, respectively, before aminophylline and by 25 +/- 7 and 13 +/- 6% after aminophylline. In a different group of six dogs, intrarenal infusion of adenosine (6 mumol/min) significantly increased RBF (32 +/- 9%) and decreased GFR (38 +/- 10%) at normal RAP. However, GFR and RBF were both well autoregulated (greater than 90% of control) at RAP values equal to or greater than 85 mmHg before and after adenosine. At RAP equal to 75 mmHg, GFR and RBF decreased by 10 +/- 5 and 7 +/- 3%, respectively, before adenosine and by 12 +/- 6 and 17 +/- 5% after adenosine. Neither aminophylline nor adenosine attenuated the elevations in plasma renin activity associated with reductions in RAP. These data fail to provide evidence that adenosine is an important factor in autoregulation of GFR and RBF during acute reductions in RAP within the autoregulatory range. PMID- 2983570 TI - Evidence for mitochondrial origin of the HCO3(-)-ATPase in brush border membranes of rat proximal tubules. AB - High HCO3(-)-ATPase activity is known to exist in mitochondria of renal tubular cells. In brush border membrane (BBM) preparations of proximal tubules such an anion-stimulated enzyme was also found. However, these preparations always contained mitochondrial markers. The putative localization and the role of this ATPase in BBM is still controversial. Some authors consider the HCO3(-)-ATPase in the BBM to be a mitochondrial contamination; others attribute to this ATPase a key role in H+ transport in the proximal tubule. To reinvestigate this problem, BBMs from rat kidney cortex were isolated by a simple, rapid (1.5-h) Ca2+ precipitation method, yielding a BBM fraction enriched 12.4-fold with respect to the marker enzyme leucine aminopeptidase (LAP). There was no basolateral Na+-K+ ATPase and no mitochondrial succinate dehydrogenase detectable. Cytochrome c oxidase was drastically reduced to 7 +/- 1% of that observed in the homogenate (TH). The activity of HCO3(-)-ATPase in the BBM fraction was 19 +/- 4 IU/g protein, i.e., 27% that of the homogenate. As sonication of the TH exclusively increases the activity of HCO3(-)-ATPase, its relative activity was 7.5% and thus equal to that of the mitochondrial marker. In many BBM preparations no HCO3(-) ATPase was detectable. In those BBM preparations in which traces of HCO3(-) ATPase were found, this activity coincided with that of cytochrome c oxidase in the respective preparation. There was a constant activity ratio of cytochrome c oxidase/HCO3(-)-ATPase in the TH, BBM, and pellet 1. The activity of HCO3(-) ATPase in BBM did not depend on the activity of LAP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983571 TI - Role of alpha-receptors in estrogen-induced vasodilation in nonpregnant sheep. AB - Estradiol-17 beta (E2) produces vasodilation in several systemic vascular beds, but most extensively in the nonpregnant uterus. It has been postulated that E2 induces this vasodilation via blockade of vascular alpha-adrenergic receptors. This hypothesis was tested in six chronically instrumented, nonpregnant sheep by comparing the systemic and uterine hemodynamic responses to intravenous E2, to an alpha-adrenergic receptor blocker, phentolamine, and to both agents given together. Uterine blood flow (UBF) increased significantly after E2 administration, from 20 +/- 7 to 233 +/- 37 (SE) ml/min. In contrast, phentolamine had no detectable effect on UBF or on the UBF response to E2 when both were given together. Similar contrasting responses were observed in the effects of E2 and/or phentolamine on the systemic vasculature. When responses to alpha-agonists were evaluated, there was no evidence of alpha-blockade following E2 despite the substantial vasodilation; in contrast, alpha-blockade was present during phentolamine administration when no vasodilation was noted. Therefore, we conclude that E2-induced vasodilation in chronically instrumented sheep is not mediated through blockade of vascular alpha-adrenergic receptors. PMID- 2983572 TI - Electrophysiological and morphological characterization of neurons in stellate ganglion of cats. AB - We have studied the general morphology of cat stellate ganglion cells in relation to the synaptic input that each neuron receives. Horseradish peroxidase (HRP) was injected intracellularly into single neurons of the isolated cat stellate ganglia. Neurons of the stellate ganglion receive synaptic information from central and peripheral nerves. Electrical stimulation of the preganglionic nerves (T3 ramus), and postganglionic stimulation of the ventral or dorsal ansa subclavia, evoked graded excitatory responses that led to the discharge of one or more action potentials. The neurons receiving synaptic input from preganglionic and postganglionic nerves have a complex dendritic morphology. These neurons were located close to the postganglionic nerves and had an axon emerging from these nerves. Other neurons located closer to the preganglionic nerves had no identifiable axons leaving the ganglion and could not be excited antidromically by electrical stimulation. These neurons appear to be interneurons. These results indicate that neurons of the cat stellate ganglion are organized in a complex fashion that could be important in the integrative properties of these neurons. PMID- 2983574 TI - Dynamic model of gonadotropin-releasing hormone receptor function. AB - Functioning gonadotropin-releasing hormone (GnRH) receptor is visualized as an aggregate of identical subunits (not all always functional) with the aggregate usually transformed into at least four successive structurally distinct receptor assemblies. Receptor protein, hormone molecule(s), and carrier(s) are main components of each functional subunit. During the normal lifespan of the functional subunit, each carrier is responsible for delivery of a unit amount of product, per unit time, to the cell surface (ratio between functional carrier and bound hormone, 1:1). Association of hormone with the receptor protein is essential, not only for the initial formation of the functional subunits but also for subsequent conformational changes that are in turn essential for formation of the aggregate only, or later (in the presence of sufficiently high GnRH concentrations) for a successive formation of a family of receptor assemblies (occurring one at a time). The successive assemblies differ from the aggregate by being more stable and from one another by increasing GnRH binding affinity and apparent capacity. They resist stimulation during protracted decay (desensitization). PMID- 2983573 TI - Role of plasma epinephrine in fasted exercising rats. AB - We have investigated the physiological role of the marked increase in plasma epinephrine that occurs in fasted exercising rats. Fasted adrenodemedullated (ADM) rats show a marked reduction in endurance run times compared with sham operated (SO) controls. After running for 30 min at 21 m/min up a 10% grade, ADM rats' blood glucose was 2.9 +/- 0.1 mM vs. 4.3 +/- 0.2 mM in SO rats. At the same time, blood lactate was 3.0 +/- 0.2 mM in SO rats compared with 1.0 +/- 0.1 mM in ADM rats. Glycogenolysis was impaired in ADM rats in the fast-twitch white region of the quadriceps, lateral gastrocnemius, and soleus muscles but not in the fast twitch red region of the quadriceps muscle. Hepatic adenosine 3',-5'-cyclic monophosphate was increased to the same extent in ADM and SO rats during exercise. Infusion of epinephrine into ADM rats during exercise corrected the hypoglycemia, restored lactate to normal, and stimulated glycogenolysis in soleus, white quadriceps, and lateral gastrocnemius muscles. Epinephrine dependent glycogenolysis in contracting type I and noncontracting type II muscle fibers apparently provides essential quantities of lactate for hepatic gluconeogenesis in fasted exercising rats. PMID- 2983575 TI - ACTH levels during cortisol suppression. PMID- 2983576 TI - An outbreak of foodborne hepatitis A showing a relationship between dose and incubation period. AB - Seven of eleven persons who attended a picnic developed symptoms of hepatitis A, two and a half to five weeks later. The epidemiologically implicated source was tuna salad, served in sandwiches and prepared by the index case. The number of sandwiches consumed was inversely related to the incubation period. This phenonemon may explain some of the variability of incubation periods for hepatitis A. PMID- 2983577 TI - Fiber intake and childhood appendicitis. AB - Parents of 135 children with appendicitis and of 212 comparison children were interviewed about their children's diet. Children in the upper two quartiles of fiber intake were estimated to have a 30 per cent lower risk of appendicitis than children in the lowest quartile. Estimated risk of appendicitis decreased as monthly intake of whole-grain foods increased. Children 7 to 18 years of age who had an intake of whole-grain foods in the upper fiftieth percentile were estimated to have a 50 per cent lower risk of appendicitis. This reduction in risk was not observed in the group of children less than 7 years of age. PMID- 2983578 TI - Laryngeal granular cell tumor in children. AB - Laryngeal granular cell tumor in childhood is rare. This article describes its occurrence in a 14-year-old girl. A review of the literature revealed five other cases. Clinical, endoscopic, histopathologic, and therapeutic considerations are discussed and compared with those in the adult form of granular cell tumor. Although this lesion is rare, awareness of it is necessary to avoid overtreatment. PMID- 2983579 TI - Benign dermal cylindroma of the external auditory canal: a clinicopathological report. AB - Considerable confusion exists regarding the nature of benign dermal cylindroma of the external auditory canal. In the past, authors have used the term cylindroma to refer to malignant lesions such as adenoid cystic carcinoma. Benign dermal cylindroma is a specific clinical entity consisting of benign tumors arising from the epidermal appendages; however, whether they are apocrine- or eccrine-derived is not conclusively known. Six per cent are reported to be related to the ear. Histologically they are characterized by cords of basophilic cells, hyaline material, and cystic duct-like structures. Rarely, cases of malignant degeneration are reported. However, for the most part, these lesions are benign and should be treated with complete local excision. Radiotherapy may be useful in treating massive lesions not amenable to surgery. PMID- 2983581 TI - The neuromuscular transmission monitor. A clinical assessment and comparison with a force transducer. AB - The neuromuscular transmission monitor (NTM) is a new device for measuring neuromuscular transmission during anaesthesia. We have assessed its use in comparison with a force transducer and found for single twitch and train-of-four modes that there was a strong positive correlation between the two methods. This device appears reliable in use and provides a satisfactory alternative to conventional neuromuscular monitoring. PMID- 2983580 TI - Allergen-induced release of sulphidopeptide leukotrienes (SRS-A) and LTB4 in allergic rhinitis. AB - Leukotrienes are a recently discovered group of arachidonic acid-derived lipid mediators. Using radioimmunoassay and high pressure liquid chromatography (HPLC), we have identified the SRS-A sulphidopeptide leukotrienes (LTC4, LTD4 and LTE4) in nasal washings from patients with allergic rhinitis who underwent nasal challenge with specific allergen. Smaller, but significant, amounts of LTB4 were also detected. The concentrations of nasal leukotrienes were directly related to the dose of allergen, and were recovered in washings in a time-dependent fashion after challenge. When the patients were subjected to methacholine nasal challenge on a control day, we found only negligible amounts of either the sulphidopeptide leukotrienes or LTB4. These findings support the view that LTC4, LTD4 and LTE4 might contribute to the pathogenesis of allergic rhinitis as a result of their recognized effects on mucous hypersecretion and vasopermeability, and that the potent chemoattractant LTB4 might be involved in the subsequent infiltration of inflammatory cells. PMID- 2983582 TI - Anaesthetic management of glucagonoma. AB - Alpha cell tumours of the pancreatic islets of Langerhans are rare. The glucagonoma syndrome is caused by excess glucagon secretion from such a tumour. Physiologically, glucagon is important in the control of the homeostatis of glucose and certain amino acids. Pharmacologically, it has been used to treat heart failure. Problems with both glucose homeostasis and myocardial function could, therefore, theoretically be anticipated following resection of a glucagonoma. This paper describes the peri-operative management of such a case, where, despite measured changes in glucagon, no problems of this nature were encountered. PMID- 2983583 TI - Thumb paraesthesia after neuromuscular twitch monitoring. AB - Nine patients, who had operations for ear and nose pathology and developed left thumb paraesthesia after neuromuscular twitch monitoring with the Gould Statham UC3 force displacement transducer are described. The pathophysiology and prevention of the complication is discussed. PMID- 2983584 TI - [Adrenoreceptors]. AB - With the development of techniques for the qualitative and quantitative assessment of receptor function and knowledge of the biological responsiveness of neurotransmitter-mediated pathways, it is now quite clear that the response of a patient to a drug does not only involve the concentration of the drug in blood and tissue. Number and function of receptors are also important factors. A perturbation in which the receptor number is elevated is called "up-regulation", whereas "down-regulation" refers to the uncoupling between receptor and effector and the consecutive decrement in the receptor concentration. In general, there is an inverse relationship between the ambient concentration of the agonist and the number of its receptors and, therefore, the sensitivity of the target organ. The demarcation between alpha- and beta-adrenergic receptors has long been appreciated. Recent advances in the understanding of adrenergic receptors have led to the subdivision of beta-receptors; beta 1-adrenoceptors mediate the stimulation of rate and force of cardiac contraction and stimulate lipolysis. beta 2-adrenoceptors mediate smooth muscle relaxation and facilitate glycogenolysis and the release of insulin, glucagon and renin. The alpha adrenergic receptors may also be divided into two subgroups. The alpha 1 adrenoceptors are postsynaptic located and facilitate smooth muscle constriction. Presynaptic located alpha 2-adrenoceptors mediate feedback inhibition of norepinephrine-release, while postsynaptic alpha 2-adrenoceptors facilitate smooth muscle contraction in selected vascular beds and stimulate the inhibition of various metabolic processes (insulin and renin secretion, lipolysis). The stage is now set for the application of the new knowledge of receptor function and regulation to the advancement of the practice of anaesthesia and intensive care. PMID- 2983585 TI - [Sodium bicarbonate administration in cardiopulmonary resuscitation. Results of an animal experimental study]. AB - One minute following electrically induced ventricular fibrillation, 12 anaesthetized pigs (hybrids between German and Belgian pedigree swine of 29 kg average body weight) were resuscitated for 30 min with a mechanical thorax compressor and ventilator. Six animals were given 1 mval sodium bicarbonate per kilogram body weight via a central venous line during the first ten minutes of resuscitation, and then 0.5 mval/kg body weight during both of the remaining ten minute intervals. The remaining 6 animals served as control group. In the control group constant volume ventilation led to a fall in the arterial partial pressure of CO2 because perfusion was diminished in relation to ventilation and despite the slowly developing metabolic acidosis this initially led to an increase in pH. At the end of the period of observation an arterial base deficit of 10 mmol/l was measured. Infusion of sodium bicarbonate led to a metabolic alkalosis and to an arterial base excess which at 10 min reached a maximum value of 13.8 mmol/l. In the bicarbonate group serum osmolality rose as a result of the sodium application from 293-334 mosm/l. In contrast, the osmolality of the control group only rose to 309 mosm/l. PMID- 2983587 TI - Nitrous oxide acts directly at the mu opioid receptor. PMID- 2983586 TI - Left ventricular performance monitored by radionuclide cardiography during induction of anesthesia. AB - Radionuclide cardiography with 99mTc-labeled erythrocytes was carried out in three different studies comprising 20 female patients without heart or lung diseases. Left ventricular ejection fraction (LVEF) and other hemodynamic variables were measured immediately before and during induction of anesthesia (thiopental, N2O/O2, succinylcholine, laryngoscopy + oral intubation, halothane). In study 1, serial measurements of LVEF, left ventricular volume, and derived variables were obtained by gamma camera in seven patients using 3-min sampling periods. In Studies 2 and 3, LVEF was monitored serially in seven and six patients, respectively, by a portable, nonimaging probe (nuclear stethoscope) at 1-min intervals or less. The induction period was prolonged to last 24 min in studies 1 and 2, against 9 min in study 3. In studies 1 and 2 there was an increase in blood pressure and heart rate after thiopental and after laryngoscopy and intubation. In study 3 a similar increase was observed after intubation. In the gamma camera study LVEF decreased from 0.72 to 0.53 after thiopental, with no further decrease during intubation. This decrease was accompanied by an increase in end-systolic volume and a decrease in the ratio: systolic cuff pressure/end systolic volume, whereas end-diastolic volume and cardiac index remained unchanged. In the nuclear stethoscope studies, LVEF decreased both after thiopental and after intubation, in study 2 from 0.68 to 0.38 and from 0.53 to 0.41, respectively; in study 3 from 0.69 to 0.53 and from 0.57 to 0.44, respectively. Our observation, in healthy, female individuals, provide an impetus for further noninvasive radionuclide studies during anesthesia in patients with cardiovascular disease. PMID- 2983588 TI - The CDC guidelines for prevention and control of nosocomial infections: view from the trenches. PMID- 2983589 TI - Hypersensitivity pneumonitis. PMID- 2983590 TI - Tuberculosis drugs--old and new. PMID- 2983591 TI - Alveolar macrophage function and inflammatory stimuli in smokers with and without obstructive lung disease. AB - To explore possible cofactors in the development of chronic obstructive pulmonary disease (COPD) in smokers, we performed bronchoalveolar lavage in 6 smokers with normal pulmonary function, 6 smokers with COPD (FEV1/FVC less than or equal to 65%) matched for smoking history and age, and 9 age-matched nonsmoking control subjects. Elastase release by macrophages from smokers with COPD was significantly higher (p less than 0.016) than was elastase release by macrophages from normal smokers. There were no differences between chemoattractiveness of alveolar macrophage supernatants for one person's polymorphonuclear leukocytes among the groups of smokers and there was no detectable C5/C5a in these supernatants (limit of detection of C5a greater than 1 ng/ml). There were no significant differences in numbers or species of bacteria in aerobically and anaerobically cultured bronchial brushings. There was no difference in alveolar macrophage superoxide anion release with particulate or membrane-perturbing stimuli for the smokers. Alveolar macrophages from the 3 groups of subjects had similar limited microbicidal ability for the obligate intracellular protozoan, Toxoplasma gondii, and similar numbers of elastase receptors and affinity for elastase. PMID- 2983592 TI - The inherited association of interstitial lung disease, hypocalciuric hypercalcemia, and defective granulocyte function. AB - The history and pulmonary histopathology of 3 siblings, presenting with the association of idiopathic interstitial lung disease, hypocalciuric hypercalcemia, and an intrinsic defect in granulocyte function are described. Prospective examination of 40 family members indicated that the 3 abnormalities are inherited according to an autosomal dominant pattern with, however, a variable penetration. Lung biopsies in the index cases revealed an interstitial infiltration of inflammatory cells and aggregates of conchoid bodies surrounded by multinucleated giant cells. Bronchoalveolar lavage was performed in 11 subjects and often showed an elevated cell recovery and abnormal cell distribution indicative of active alveolitis. In several subjects, multinucleated giant cells were found. The diffusing capacity (in percent predicted) showed a significant decrease with age, independent of smoking habits. The hypocalciuric hypercalcemia was unaffected by steroids or parathyroidectomy. It was not associated with abnormal levels of parathyroid hormone, calcitonin, 25-hydroxy-vitamin D3, 1,25-dihydroxy-vitamin D3, or angiotensin-converting enzyme. PMID- 2983593 TI - Immunoglobulin G antibodies to lymphadenopathy-associated virus in differently treated French and Belgian hemophiliacs. AB - Immunoglobulin G antibodies to lymphadenopathy-associated virus have been detected in two groups of French hemophiliacs and in one group of Belgian hemophiliacs, whose mode of treatment differed. Seropositivity was more frequent (58.9%) in patients heavily transfused with blood products of French and foreign origin than in less frequently transfused persons (10.3%). The Belgian group, treated only with local products, showed the lowest frequency of seropositivity (3.4%). In healthy French controls, 1 of 330 had antibody to the virus. The results indicate transmission of lymphadenopathy-associated virus via blood derived products. PMID- 2983594 TI - Extrahepatic biliary obstruction caused by small-cell lung cancer. AB - Twelve patients with small-cell lung cancer seen during a 30-month period had jaundice at diagnosis. Five patients had a pancreatic metastasis resulting in extrahepatic biliary obstruction, and seven had diffuse hepatic metastases without extrahepatic obstruction. All patients with pancreatic masses had complete (or nearly complete) resolution of jaundice and abdominal pain within 3 weeks of starting chemotherapy. Patients with extensive liver metastases usually remained icteric in spite of intensive treatment. Three patients with pancreatic metastases survived more than 12 months after the institution of therapy. No patient presenting with jaundice caused solely by hepatic metastases survived beyond 8 months. Small-cell lung cancer can present with jaundice due to diffuse hepatic parenchymal involvement, which is associated with a poor prognosis, or as a result of extrahepatic biliary obstruction, which has potential for rapid palliation and prolonged survival. PMID- 2983595 TI - Primary Sjogren's syndrome after infectious mononucleosis. AB - A healthy young woman developed primary Sjogren's syndrome after protracted infectious mononucleosis. The diagnosis of primary Sjogren's syndrome was supported by histologic evidence of sialadenitis in labial salivary glands, rheumatoid factor, hypergammaglobulinemia, the HLA-B8 phenotype, and a high titer antibody to the anti-La (SS-B) nucleoprotein that co-precipitated the small ribonucleic acids encoded by Epstein-Barr virus, EBER 1 and EBER 2, as well as "host" RNA. There was strong humoral immunity to the Epstein-Barr nuclear and capsid antigens, but weak T-lymphocyte-mediated cytotoxicity to Epstein-Barr transformed lymphoblasts, anergy to antigens used to elicit delayed-type hypersensitivity, and a low T-helper/T-suppressor cell ratio. The series of events initiated by infectious mononucleosis was attributed to a genetic defect in the immune response. Association of viral RNA with the La nucleoprotein resulted in a break in immunologic tolerance via a T-cell bypass effect with induction of anti-La (SS-B) by polyclonally activated B lymphocytes leading to autoimmune sialadenitis. PMID- 2983596 TI - Oncogenic osteomalacia. PMID- 2983598 TI - [Premature development of the breasts. Value of utero-ovarian evaluation by pelvic ultrasound]. PMID- 2983597 TI - [Isolated glucocorticoid deficiency manifested at birth (Migeon syndrome). Review of the literature apropos of a case]. PMID- 2983599 TI - A system for long term perfusion of arterial cannulae. PMID- 2983600 TI - Desaturation of 14C linoleic acid by the rat fetus. PMID- 2983601 TI - Dermatomal somatosensory evoked potentials in unilateral lumbosacral radiculopathy. AB - We examined scalp-recorded somatosensory evoked potentials (SSEPs) to electrical stimulation of the peroneal nerves and to stimulation in the L5 and S1 dermatomes in 19 patients with unilateral radiculopathies involving these segments. For the dermatomal studies at least two trials of 512 responses were recorded from the vertex with reference to both the midfrontal and contralateral parietal electrodes, using an averaging technique. Findings on the symptomatic and asymptomatic sides were compared in each patient. We found that peroneal SSEPs were normal in all patients. Dermatomal SSEPs correctly identified the lesion in 5 patients. In 1 patient dermatomal SSEPs lateralized the lesion correctly but localized it to the adjacent root. In 10 cases dermatomal SSEPs gave misleading information, indicating an abnormality on the asymptomatic side in 1 patient and no abnormality in 9. In the remaining 3 patients, both SSEPs and radiological contrast studies failed to identify any lesion, although the radiculopathy was confirmed by electromyography. These findings raise doubt about the ultimate utility of these evoked potential techniques in the evaluation of patients with suspected radiculopathies. PMID- 2983602 TI - Herpes simplex virus type 2 encephalitis in two homosexual men with persistent lymphadenopathy. AB - Within a 5-month period, 2 homosexual men with persistent lymphadenopathy developed clinical findings consistent with herpes simplex virus (HSV) encephalitis. These signs included abrupt change in mental status, seizures, cerebrospinal fluid pleocytosis, and localized electroencephalographic abnormalities showing temporal lobe involvement. Initial computed tomographic scans were unremarkable. Treatment with adenine arabinoside was instituted and temporal lobe biopsies were performed. Although virus-specific antigens were detectable in only 1 patient, cultures of biopsy tissue from both patients yielded HSV type 2 organisms. Spiking fevers persisted and the patients failed to improve, prompting administration of acyclovir. Both patients recovered gradually after their second course of antiviral therapy and survived with severe neurological deficits. These patients should raise concerns about an increased incidence of type 2 encephalitic illness among homosexual men with persistent lymphadenopathy or acquired immune deficiency syndrome. In addition, the importance of using HSV type 2 antibody in the immunofluorescence test of brain biopsy tissue for rapid diagnosis of the disease is emphasized. PMID- 2983603 TI - Neuronal intranuclear inclusion disease in two adult siblings. PMID- 2983604 TI - Cellular mechanisms of learning, memory, and information storage. AB - In Table 1, we summarize what is convincingly demonstrated to date for the major vertebrate and invertebrate model systems attempting to elucidate cellular mechanisms of associative learning. Two major concerns are the adequacy of the behavioral demonstrations and the completeness and extent of the accompanying neurophysiology. In addressing the issue of behavior, it is important to define clearly which criteria are both necessary and sufficient to infer the involvement of an associative-learning process. Similarly, it is also important to distinguish among those primary characteristics of associative learning in general, and those secondary or tertiary features that serve to define various subclasses. In our view, it would be unreasonable to require that any given preparation exhibit all the defining features of classical conditioning, for example, in order to qualify as a "legitimate" instance of associative learning. This is especially true if the goal is to understand the more general, rather than the specific, mechanisms involved in associative learning. Hence, we emphasize the following as primary features of learned behavior: pairing specificity, stimulus specificity, long-term retention (arbitrarily defined as lasting for at least 24 hr), a moderate degree of reversibility by subsequent experience (e.g. extinction), and demonstrations that nonassociative-learning processes cannot account for features a-c. Where appropriate, we also identified other interesting features of the learned behavior. It is apparent from the table that a major unresolved issue for most of the preparations is the extent to which the behavioral changes are exclusively associative. This is no less true for the vertebrate preparations than it is for the invertebrates. The clearest example of an exclusively associative behavioral change is the rabbit NMR. The learning produced changes in the invertebrate preparations were all shown, to varying degrees, to be pairing specific. Yet a major unresolved issue is the degree to which apparent examples of associative-learning reflect complex interactions among basically nonassociative-learning processes. The core issue is really quite simple: Does the associative training procedure result in the acquisition of new or qualitatively different behavior; and is there a strict requirement for an associative relation? In addressing the adequacy of the neurophysiological analyses, the major issue is that of localization. Logically, there are two components to this.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983605 TI - Production of D- and L-xylulose by mutants of Klebsiella pneumoniae and Erwinia uredovora. AB - D-Xylulose and L-xylulose were produced biologically by the oxidation of a corresponding pentitol. A Klebsiella pneumoniae mutant was constructed for the oxidation of D-arabitol to D-xylulose. This mutant constitutively synthesized the D-arabitol permease system and D-arabitol dehydrogenase but was unable to produce the D-xylulokinase of the D-arabitol pathway or the D-xylose isomerase and D xylulokinase of the D-xylose pathway. An Erwinia uredovora mutant which constitutively synthesized a novel xylitol-4-dehydrogenase but could not synthesize L-xylulokinase was used for the oxidation of xylitol to L-xylulose. Washed cell suspensions of either mutant incubated with 0.5% pentitol would oxidize 60 to 65% of the pentitol to the corresponding ketopentose in 18 h and excrete the ketopentose into the medium. Ketopentoses were rapidly purified from the remaining pentitol by hydroxyl affinity chromatography. PMID- 2983606 TI - A temperate phage of Streptomyces azureus. AB - A new phage, SAt1, was isolated from soil on Streptomyces azureus ATCC 14921. This phage was able to lysogenize S. azureus. The percentage of lysogenic responses was ca. 10%. Electron microscopic observation showed that this phage belonged to group B of Bradley's morphological classification. The molecular mass of SAt1 DNA was ca. 24 megadaltons. The guanine-plus-cytosine content and the density of SAt1 DNA were ca. 71% and 1.724 g/cm3, respectively. A cleavage map of SAt1 DNA was constructed with restriction endonucleases BanIII, BglII, HindIII, and XbaI. Furthermore, some other characteristics of this phage were investigated. PMID- 2983607 TI - Detection of false-positives among total and fecal coliform counts by factorial analysis of correspondence. AB - Application of an analysis of correspondence to the biochemical characteristics of total and fecal coliforms isolated in the Ivory Coast permitted us to separate two small clusters of isolates different from the main clusters, which included isolates from human and animal feces. The isolates grouped in the small clusters were from water samples. An analysis of the biochemical characteristics which permitted the segregation of the "water-specific" isolates from the main clusters indicates that water-specific total coliforms were citrate positive, indole negative, and amygdaline positive. Water-specific fecal coliforms were either citrate positive, indole negative, amygdaline positive, and inositol negative or indole negative, amygdaline positive, and inositol positive. Any isolates not fitting the above patterns could be considered of fecal origin. If this observation is confirmed under temperate climates and for a greater number of isolates, these simple tests could be used to confirm the fecal origin of coliforms. PMID- 2983608 TI - Adsorption of viruses to charge-modified silica. AB - The purpose of this study was to provide a clearer understanding of virus adsorption, focusing specifically on the role of electrostatic interactions between virus particles and adsorbent surfaces. The adsorption of poliovirus 1, reovirus types 1 and 3, and coliphages MS-2 and T2 to colloidal silica synthetically modified to carry either positive or negative surface charge was evaluated. Adsorption experiments were performed by combining virus and silica in 0.1-ionic-strength buffers of pH 4.0, 6.4, and 8.5. Samples agitated for specified adsorption periods were centrifuged to pellet adsorbent particles plus adsorbed virus, and the supernatants were assayed for unadsorbed virus. All viruses adsorbed exclusively to negatively charged silica at pH values below their isoelectric points, i.e., under conditions favoring a positive surface charge on the virions. Conversely, all viruses adsorbed exclusively to positively charged silica at pH values above their isoelectric points, i.e., where virus surface charge is negative. Viruses in near-isoelectric state adsorbed to all types of silica, albeit to a lesser degree. PMID- 2983609 TI - A multifunctional calmodulin-stimulated phosphatase. AB - This review summarizes current knowledge concerning structure-function, substrate specificity, localization, and regulatory properties of calcineurin. Calcineurin is composed of two nonidentical subunits, one of which is responsible for catalytic activity and calmodulin binding while the other subunit contains four high-affinity Ca2+-binding sites. The enzyme possesses calmodulin-stimulated and metal ion-dependent phosphatase activity toward several nonprotein and phosphoseryl-, phosphothreonyl- and phosphotyrosyl-containing protein substrates. These recent results suggest that the protein may play a multifunctional role in interactions between the Ca2+/CaM second messenger system and other second messenger systems. PMID- 2983610 TI - Partial characterization of nuclear binding sites for retinol delivered by cellular retinol binding protein. AB - Retinol (vitamin A alcohol), which plays an important role in the differentiation of epithelia, can be transferred to chromatin in vitro. Rat liver chromatin can accept retinol in a specific and saturable manner only when the retinol is presented as a complex with cellular retinol-binding protein (CRBP). A partial characterization of the nuclear components responsible for accepting retinol is reported here. A preparation of solubilized chromatin isolated from liver nuclei was able to accept retinol from its complex with CRBP as described previously for nuclei and chromatin. The binding of retinol to chromatin was noncovalent. However, chromatin prepared from nuclei which were incubated with DNase I or micrococcal nuclease did not accept retinol specifically. Chromatin in the form of mono and dinucleosomes also did not accept retinol. However, treatment of nuclei with RNase did not affect the specific binding of retinol. Furthermore, it has been found that retinol was not transferred to purified double or single stranded DNA. These results are interpreted to indicate that the transfer of retinol to specific nuclear binding sites requires a higher order of chromatin structure than that occurring in nucleosome preparations. PMID- 2983611 TI - Regulation of ornithine aminotransferase by cyclic AMP and glucose in primary cultures of adult rat hepatocytes. AB - Hepatic ornithine aminotransferase (EC 2.6.1.13) (OAT) is a mitochondrial matrix enzyme that plays a role in amino acid catabolism and in gluconeogenesis. In rats, the synthesis of hepatic OAT is regulated by glucagon, dietary protein, and glucose. Serum-free primary cultures of adult rat hepatocytes were used to demonstrate that glucagon, cyclic AMP, and glucose are able to alter OAT synthesis by a direct action on hepatocytes. The rates of OAT synthesis were measured by immunoprecipitation of pulse-labeled OAT with an affinity-purified monospecific antibody. Ten hours after cyclic AMP addition to the culture medium, the relative rate of OAT synthesis reached a peak value that was six- to eightfold above the control rate. OAT activity accumulated more slowly, reaching a level that was approximately threefold above the control by 24 h. The inclusion of glucose in the culture medium inhibited the increases in OAT synthesis and activity in a dose-dependent manner. Although synthesized as a precursor (pOAT), no pOAT was detected under control, induced, or carbohydrate-inhibited conditions; this suggests that pOAT processing may not be a regulatory site of OAT expression. By following the loss of labeled OAT, a half-life of 34 h in these cultures under all of the above conditions was observed. Regulation of OAT levels in cultured hepatocytes appears to be achieved primarily through changes in the rate of OAT synthesis. PMID- 2983612 TI - Modified cation activation of the (Na+K)-ATPase following treatment with thimerosal. AB - Treatment of the Na,K-ATPase enzyme, isolated from canine renal outer medulla, with thimerosal (ethylmercurithiosalicylate) resulted in significant inhibition of the overall Na,K-ATPase with only slight, if any, inhibition of the Na-ATPase and ATP:ADP exchange activities. The K-stimulated PNPPase activity was stimulated [see G. R. Henderson and A. Askari (1977) Arch. Biochem. Biophys. 182, 221-226]. Examination of the Na dependence of the ATPase and ATP:ADP exchange activities revealed an Na-independent, ouabain-sensitive activity that was inhibited by Na in the range 0-10 mM. At greater than or equal to 10 mM concentration of Na the treated and modified enzymes showed similar activities. The apparent affinity of the modified enzyme for ATP in the presence of 100 mM Na was the same as that of the untreated enzyme (0.2-0.3 microM). In the absence of Na, the modified enzyme hydrolyzed ATP with a relatively low affinity (about 120 microM). The enhancement of p-nitrophenylphosphatase (PNPPase) activity measured in the presence of K ions was due to the appearance of K-independent, ouabain-sensitive PNPP activity. The modification was without major affect on the apparent affinity of the enzyme for K ions in the PNPPase activity. Treatment of the thimerosal-modified enzyme with dithiothreitol removed (or greatly reduced) the cation-independent, ouabain sensitive activities and the Na,K-ATPase activity returned. Modification of a set of enzyme -SH groups in the Na,K-ATPase enzyme made it able to hydrolyze ATP in the absence of Na ions and PNPP in the absence of K ions. The -SH groups modified by thimerosal are evidently critical to the major dephosphoenzyme conformational changes but are not involved in the major transport conformational change between phosphoenzymes. PMID- 2983613 TI - Ubisemiquinone is the electron donor for superoxide formation by complex III of heart mitochondria. AB - Much evidence indicates that superoxide is generated from O2 in a cyanide sensitive reaction involving a reduced component of complex III of the mitochondrial respiratory chain, particularly when antimycin A is present. Although it is generally believed that ubisemiquinone is the electron donor to O2, little experimental evidence supporting this view has been reported. Experiments with succinate as electron donor in the presence of antimycin A in intact rat heart mitochondria, which contain much superoxide dismutase but little catalase, showed that myxothiazol, which inhibits reduction of the Rieske iron sulfur center, prevented formation of hydrogen peroxide, determined spectrophotometrically as the H2O2-peroxidase complex. Similarly, depletion of the mitochondria of their cytochrome c also inhibited formation of H2O2, which was restored by addition of cytochrome c. These observations indicate that factors preventing the formation of ubisemiquinone also prevent H2O2 formation. They also exclude ubiquinol, which remains reduced under these conditions, as the reductant of O2. Since cytochrome b also remains fully reduced when myxothiazol is added to succinate- and antimycin A-supplemented mitochondria, reduced cytochrome b may also be excluded as the reductant of O2. These observations, which are consistent with the Q-cycle reactions, by exclusion of other possibilities leave ubisemiquinone as the only reduced electron carrier in complex III capable of reducing O2 to O2-. PMID- 2983614 TI - Photoinduced reactions of anthraquinone antitumor agents with peptides and nucleic acid bases: an electron spin resonance and spin trapping study. AB - The photoexcitation (lambda = 313 +/- 10 nm) of adriamycin, daunomycin, and mitoxantrone in the presence of peptides or pyrimidine nucleic acid bases was investigated. In air-saturated and air-free solutions, peptides are decarboxylated by the photoexcited drug molecules. The decarboxylation reactions were shown to occur specifically at the C-terminal amino acid of the peptide. The decarboxylated peptide radicals were spin-trapped using 2-methyl-2-nitrosopropane (MNP) and identified by electron spin resonance (ESR). In air-free solutions, nucleic acid bases are oxidized by the photoexcited drug molecules predominantly generating C(5)-carbon-centered radicals in the pyrimidine rings of uracil, cytosine, and thymine. However, spin adducts of MNP and thymine were also obtained at the N(1) or N(3) positions of the pyrimidine ring. In air-saturated adriamycin and daunomycin solutions, the spin adducts of MNP with uracil or thymine are similar to those obtained following hydroxyl radical reactions with these pyrimidines. This suggests that in the presence of oxygen, the photoexcited adriamycin and daunomycin transfer an electron to oxygen generating the superoxide anion radicals (O2-.), which are precursors of hydroxyl radicals. O2-. was also formed when O2-saturated DNA solutions were photoirradiated (lambda = 313 +/- 10 and 438 +/- 10 nm) in the presence of adriamycin and daunomycin, indicating that the photodegradation of DNA in the presence of these drugs caused by hydroxyl radicals is mediated by dissolved oxygen. PMID- 2983616 TI - Angiomatoid malignant fibrous histiocytoma associated with marked bleeding arising in chronic radiodermatitis. PMID- 2983615 TI - Regulation of C4 photosynthesis: purification and properties of the protein catalyzing ADP-mediated inactivation and Pi-mediated activation of pyruvate,Pi dikinase. AB - Pyruvate,Pi dikinase regulatory protein (PDRP) has been highly purified from maize leaves, and its role in catalyzing both ADP-mediated inactivation (due to phosphorylation of a threonine residue) and Pi-mediated activation (due to dephosphorylation by phosphorolysis) of pyruvate,Pi dikinase has been confirmed. These reactions account for the dark/light-mediated regulation of pyruvate,Pi dikinase observed in the leaves of C4 plants. During purification to apparent homogeneity the ratio of these two activities remained constant. The molecular weight of the native PDRP was about 180,000 at pH 8.3 and 90,000 at pH 7.5. Its monomeric molecular weight was 45,000. It was confirmed that inactive pyruvate,Pi dikinase free of a phosphate group on a catalytic histidine was the preferred substrate for activation. Michaelis constants for orthophosphate and the above form of active pyruvate,Pi dikinase were determined, as well as the mechanism of inhibition of the PDRP-catalyzed reaction by ATP, ADP, AMP, and PPi. For the inactivation reaction, Km values were 1.2 microM for the active pyruvate,Pi dikinase and 52 microM for ADP. CDP and GDP but not UDP could substitute for ADP. The inactivation reaction is inhibited by inactive pyruvate,Pi dikinase competitively with respect to both active pyruvate,Pi dikinase and ADP. Both the activation and inactivation reactions catalyzed by PDRP have a broad pH optimum between 7.8 and 8.3. The results are discussed in terms of the likely mechanism of dark/light regulation of pyruvate,Pi dikinase in vivo. PMID- 2983617 TI - Fibroepithelioma of Pinkus overlying breast cancer. PMID- 2983618 TI - Papulosquamous eruption with weight loss: Necrolytic migratory erythema (glucagonoma syndrome). PMID- 2983619 TI - Metastasizing eccrine porocarcinoma developing in a sebaceous nevus of Jadassohn. Report of a case. PMID- 2983620 TI - Malignant fibrous histiocytoma arising from chronic ulcer. AB - A patient had malignant fibrous histiocytoma that developed in a chronic ulcer. Squamous cell carcinoma can develop on long-standing ulcers, but malignant fibrous histiocytoma developing in an ulcer is extremely rare. PMID- 2983621 TI - Epidermotropic urothelial carcinoma involving the glans penis. AB - A case of epidermotropic carcinoma involving the urethral meatus and the skin of the glans penis was associated with primary transitional cell carcinoma of the urinary bladder with urethral extension. The histopathologic and histochemical features were identical to those seen in extramammary Paget's disease. There was positive intracytoplasmic staining for the carcinoembryonic antigen with the use of an immunoperoxidase stain, another similarity between this neoplasm and extramammary Paget's disease. However, these staining characteristics are consistent with those of the primary bladder tumor and with staining patterns described in the literature for urothelial carcinoma. PMID- 2983622 TI - Postirradiation malignant fibrous histiocytoma. AB - A slowly growing, firm, asymptomatic tumor developed in the skin overlying the left scapula of a 72-year-old woman during a six-month period. Twenty-seven years earlier, she had radiation treatment to that site for an intraductal carcinoma of the left breast. The histopathologic diagnosis was malignant fibrous histiocytoma (MFH). The rarity of this late complication of radiation therapy is reflected by a few reports in the medical literature of cutaneous MFH as late radiation sequelae. PMID- 2983623 TI - Ultrasound and the diagnosis of Wilms' tumour. PMID- 2983624 TI - Seizures and steroids. PMID- 2983625 TI - Serum alpha interferon and lymphocyte inclusions in systemic lupus erythematosus. AB - The relationship between serum acid-labile alpha interferon and tubuloreticular inclusions within the cytoplasm of circulating lymphocytes was studied in 46 patients with systemic lupus erythematosus. Elevated levels of interferon (greater than or equal to 8 IU/ml) were found in 17 patients and lymphocyte inclusions in 35. The mean serum interferon concentration in patients with lymphocyte inclusions was significantly higher than in patients without inclusions (17.2 versus 2.4 IU/ml, p less than 0.01). Inclusions were found in 16 of 17 patients with elevated interferon and also in 19 of 29 patients without interferon (p = 0.026). In lupus, serum interferon appears to be a sufficient though not an essential marker for the presence of lymphocyte inclusions. PMID- 2983626 TI - Klippel and Trenaunay's syndrome. 768 operated cases. AB - Since 1945, we have operated on 786 patients with Klippel and Trenaunay's syndrome. Elongation of the impaired limb was invariably found while edema was present in 84%, varicose veins in 36%, and flat angiomata in 32%. Venography and surgical exploration have demonstrated malformation of the deep veins involving the popliteal vein in 51%; superficial femoral vein, 16%; both popliteal and superficial femoral veins; 29%; iliac veins, three per cent; and lower vena cava, one per cent. Good clinical results have been achieved following the surgical release of these deep veins in the lower limb. During childhood, when the difference in limb length is noteworthy, ligature of the popliteal vein of the shorter limb induces a compensating elongation. Klippel and Trenaunay's syndrome may be associated with lymphatic malformations, including lymphedema and malformation of the lymph vessels. Knowledge of the pathophysiology of these malformations of the deep veins enables a better understanding of the clinical manifestations of the condition, as well as the improved treatment of the serious vesical or rectal hemorrhage which occurs in one per cent of these patients. PMID- 2983627 TI - Abnormal thyrotropin and prolactin levels in untreated corticotropin deficiency. AB - A patient with isolated corticotropin deficiency demonstrated hyperresponsiveness of both thyrotropin and prolactin after protirelin stimulation. Following glucocorticoid replacement therapy, responses decreased to normal. Since true primary hypothyroidism may occur in association with cortisol-deficient states, it is necessary to reserve final evaluation until after glucocorticoid replacement. The finding of combined thyrotropin and prolactin hyperresponsiveness to protirelin suggests a related glucocorticoid feedback function. PMID- 2983628 TI - Obstruction of the superior vena cava after chemotherapy. AB - Superior vena cava obstruction developed a few hours after the administration of chemotherapy for small-cell lung carcinoma. The syndrome responded rapidly to dexamethasone. Although the mechanisms of the appearance and resolution of the superior vena cava obstruction in this patient remain hypothetic, there is an important therapeutic implication. PMID- 2983629 TI - Nonpersimmon gastric phytobezoars. PMID- 2983631 TI - NIMH Treatment of Depression Collaborative Research Program. Background and research plan. AB - The National Institute of Mental Health (NIMH) Treatment of Depression Collaborative Research Program (TDCRP) is the first multisite coordinated study initiated by the NIMH in the field of psychotherapy research. Three research sites, using an identical research protocol, are investigating the effectiveness of two forms of brief psychotherapy (cognitive behavior therapy and interpersonal psychotherapy) in the treatment of outpatient depression. Three training sites have trained experienced therapists in a standard fashion for each of the psychotherapies and the comparison pharmacotherapy conditions. This report presents the background of the TDCRP, the rationale for the choice of patient population and treatment conditions, and the research plan for both the training/pilot phase and the outcome study currently in progress, and discusses the potential contributions of the program to the field of psychotherapy research. PMID- 2983630 TI - Increased anxiogenic effects of caffeine in panic disorders. AB - The effects of oral administration of caffeine (10 mg/kg) on behavioral ratings, somatic symptoms, blood pressure and plasma levels of 3-methoxy-4 hydroxyphenethyleneglycol (MHPG) and cortisol were determined in 17 healthy subjects and 21 patients meeting DSM-III criteria for agoraphobia with panic attacks or panic disorder. Caffeine produced significantly greater increases in subject-rated anxiety, nervousness, fear, nausea, palpitations, restlessness, and tremors in the patients compared with healthy subjects. In the patients, but not the healthy subjects, these symptoms were significantly correlated with plasma caffeine levels. Seventy-one percent of the patients reported that the behavioral effects of caffeine were similar to those experienced during panic attacks. Caffeine did not alter plasma MHPG levels in either the healthy subjects or patients. Caffeine increased plasma cortisol levels equally in the patient and healthy groups. Because caffeine is an adenosine receptor antagonist, these results suggest that some panic disorder patients may have abnormalities in neuronal systems involving adenosine. Patients with anxiety disorders may benefit by avoiding caffeine-containing foods and beverages. PMID- 2983632 TI - Cytomegalovirus encephalitis in a patient with acquired immunodeficiency syndrome. PMID- 2983633 TI - Differential diagnosis of Kaposi's sarcoma. AB - The biopsies of all lesions clinically thought to be suspicious for Kaposi's sarcoma (KS) were reviewed over a 15-month period. A diagnosis of KS was made in 40 of 106 biopsies (38%). The cases in which a diagnosis other than KS was made included dermatofibroma, hemangioma, and scar. This second group comprised 59 of 106 cases (56%). A third group included some lesions that had an atypical vascular proliferation, but in which the changes were insufficient for a definite diagnosis of KS. The presence of abnormally shaped vessels, especially those classified as irregular, was the best single criterion to diagnose KS in its early stages. In later stages, the neoplasm assumes a nodular configuration with typical, slitlike vascular channels. At the periphery of such nodules dilated, irregularly shaped vessels similar to those of the early lesions are often seen. The histologic features which help in the diagnosis of KS from other histologic entities are reviewed. PMID- 2983634 TI - Disseminated talc granulomatosis. An unusual finding in a patient with acquired immunodeficiency syndrome and fatal cytomegalovirus infection. AB - The association of disseminated magnesium silicate talc granulomatosis and acquired immunodeficiency syndrome is reported in a male homosexual who used intravenous drugs and who died of overwhelming cytomegalovirus (CMV) infection. Autopsy findings included widespread deposition of talc crystals in the lungs, liver, lymph nodes, bone marrow, and spleen. Typical CMV inclusions were seen in the lungs, kidneys, adrenal glands, gastrointestinal tract, and right eye. There was no evidence of malignancy. Analysis of peripheral blood neutrophil function revealed impaired chemotaxis and chemokinesis, but opsonophagocytosis had remained normal. The CMV infection in the small bowel was extensive and resulted in severe destruction of the muscularis propria and neural plexi, leading to marked dilatation and persistent diarrhea. The terminal course was marked by intractable hypotension, pneumonitis, and malnutrition, which could be attributed respectively to CMV involvement of the adrenal glands, lungs, and small bowel. The etiology and possible role of systemic talc granulomatosis in the development of immunosuppressive illness is reported herein. PMID- 2983635 TI - A comparison between two commercially available activators for determining the partial thromboplastin time. AB - Activators that are used to determine the activated partial thromboplastin time should be sensitive enough to detect mild deficiencies of coagulation factors in a reproducible fashion. We compared two commercially available activators--a newly modified preparation of ellagic acid activator and a silica activator. Both reagents produced results that were reproducible over a broad range of concentrations of factors involved in the intrinsic coagulation pathway. The ellagic acid activator was more sensitive in detecting a partial deficiency of nearly all of these factors. PMID- 2983636 TI - Morphologic identity of primary tumor and axillary metastases in breast carcinoma. A quantitative study. AB - The amount of glandular differentiation was quantitated in 64 cases of primary human mammary ductal carcinoma (PR) and their axillary lymph node metastases (LNMs) to identify differences between the two groups and their effect on patient prognosis. A significant difference between a PR and any of its LNMs was uncommon (less than 5% of the cases), and variation between each PR and its LNMs was directly proportional to the amount of differentiation within the PR itself. Variance among the LNMs was proportional to that found within the PR. Each LNM usually varied as much within itself as it did from the other LNMs. Prognosis was not affected by the LNMs being less differentiated than the PR, nor vice versa. This study has provided no evidence for selection of a morphologically more aggressive tumor clone during this phase of the metastatic process, but selection by other morphologic or functional parameters is not excluded. PMID- 2983637 TI - Peripheral small-cell carcinoma of the lung resembling carcinoid tumor. A clinical and pathologic study of 14 cases. AB - We studied 14 small-cell, epithelial tumors of the lung with histologic features intermediate between typical carcinoid tumor and undifferentiated small-cell carcinoma. All of the tumors arose in the periphery of the lung beyond a segmental bronchus and were excised. Histologically, three of the tumors were low grade and 11 were high grade. The low-grade tumors had an organoid pattern comprising more than half of the area examined histologically, less than five mitoses per 10 high-power fields, individual cell necrosis, and nuclear pleomorphism of less than half of the cells. The high-grade tumors had the opposite characteristics. Distinction of these peripheral tumors from typical carcinoid tumors and from undifferentiated small-cell carcinomas is established after excision and cannot be made reliably on the findings of a transbronchial or needle biopsy. Prognosis was worse than that for typical carcinoid tumors but better than that for undifferentiated small-cell carcinomas. PMID- 2983638 TI - Small-cell (oat cell) carcinoma of the endometrium. AB - We report a case of small-cell (oat cell) carcinoma of the endometrium in a 59 year-old woman. The tumor was confined to a portion of an adenomatous polyp. It exhibited argyrophilia with Grimelius' stain and dense-core endocrine-type granules by electron microscopy. The patient was alive and well one year later. To our knowledge, this is the fourth reported case of a small-cell carcinoma of the endometrium exhibiting endocrine differentiation. PMID- 2983639 TI - Metastasis to pituitary adenoma. AB - Neoplasm to neoplasm metastasis is a medical curiosity. We report two cases of adenocarcinoma metastatic to pituitary adenoma. In both, abrupt progression of symptoms referable to the sellar region was noted and followed by the death of the patient. PMID- 2983640 TI - Lumbosacral radiculopathy, reflex sympathetic dystrophy and tarsal tunnel syndrome: an unusual presentation. AB - This case documents a patient with lumbosacral radiculopathy, reflex sympathetic dystrophy, and tarsal tunnel syndrome affecting one extremity. After lumbar laminectomy for L5-S1 radiculopathy, the patient developed foot pain diagnosed as secondary to reflex sympathetic dystrophy. Because of increasing foot pain despite long-term conservative therapy, the patient underwent further evaluation, including electrodiagnostic studies leading to the diagnosis of a superimposed tarsal tunnel syndrome. Following surgical decompression, pain decreased. Because of the unusual presentation, diagnosis and treatment of the tarsal tunnel syndrome was significantly delayed. To help understand the coexistence of these conditions, interrelating pathophysiologic mechanisms have been postulated, including the possibility that a "double-crush" phenomenon may have contributed to the development of tarsal tunnel syndrome. Earlier diagnosis and definitive treatment of the foot pain may have been possible had these mechanisms been better understood. PMID- 2983641 TI - Effects of estrogen treatment on sexual behavior in male-to-female transsexuals: experimental and clinical observations. AB - The effects of oral estrogen treatment on sexual physiology and behavior were examined in seven presurgical male-to-female transsexuals engaged in cross living. Subjects were studied prior to hormone treatment, during long-term hormone treatment, and during an experimental double-blind period in which the effects of their usual hormone regimen were compared to those of placebo during successive 4-week periods. Subjects maintained daily logs of their spontaneous erections, sexual activity (masturbation), and feelings throughout the study. Nocturnal penile tumescence was measured, using home monitors, in order to estimate estrogen-induced changes in erectile capacity. Erectile response to sexually arousing stimuli (erotic films and self-generated fantasy) was also assessed in the laboratory. Blood samples were taken at intervals for testosterone and sex-hormone-binding globulin measurements and free testosterone levels were calculated. Estrogen treatment inhibited sexual activity, spontaneous erections, and nocturnal penile tumescence. No significant effects on psychophysiological response to film and fantasy or frequency of sexual feelings were found, but the psychophysiological data were very variable. Testosterone levels were suppressed by estrogen, but not to the extent that free testosterone levels were. It appears that declining free testosterone level is associated with inhibition of spontaneous erections (during both sleep and waking) and of sexual activity, though the latter relationship is less clear. No evidence of an effect on film or fantasy-induced erections was obtained. PMID- 2983642 TI - Effects of vitamin B12 and B6 on 2,5-hexanedione-induced neuropathy. AB - The therapeutic effect of Vitamin B12 or Vitamin B6 on 2,5-hexanedione induced neuropathy in animals was examined by using our electrodiagnostic technique in order to investigate the mechanism of the development of the neuropathy. Pyridoxal phosphate and two forms of Vitamin B12 were administered to rats intoxicated by the neurotoxin for a period of 18 weeks, and the sensory and motor fiber conduction velocity and the motor distal latency of the tail nerve were periodically determined. None of the groups treated with the therapeutic drugs exhibited a definite improvement in the nerve fiber conduction velocity and motor distal latency of the rat tail nerve, as compared with those of the 2,5 hexanedione-treated controls. PMID- 2983644 TI - [Histoenzymatic characteristics of the articular cartilage in ontogeny and in osteoarthrosis deformans]. AB - Enzymo-histochemical study of the human joint cartilage in ontogenesis showed a high 5-nucleotidase activity at all stages of human development. Alkaline phosphatase was detectable only in the zones of enchondral ossification of the joint cartilage and in the ossification "nuclei", while in adults only in the chondrocytes over the ossification line. Osteocytes of the subchondral bone contain neither 5-nucleotidase nor alkaline phosphatase, the latter being detected in the endosteal cells only. The damage of the joint cartilage in the osteoarthrosis results in the redistribution of the osteo- and chondrogenesis enzymes with their localization in the zones of the regenerative reconstruction of the subchondral bone tissue. PMID- 2983643 TI - [Reparative regeneration of rat liver after treatment with bee venom]. AB - After 70% of the rat liver volume has been resected, certain stimulating effect of a single administration of bee venom (1 mg/kg) on hepatocyte proliferation is revealed. In 4 hours the amount of binuclear hepatocytes increases, and in 22-24 hours increases their mitotic activity. The effect of bee venom is explained as activation of the adenilate cyclase system by means of prostaglandins and adrenal hormones. PMID- 2983645 TI - [Synovial chondromatosis]. AB - Synovial chondromatosis is characterized by formation of cartilaginous nodes, possibly with ossification, in the synovial membrane of the joints, tendons, and vagina. The description of a case of synovial chondromatosis located in the knee joint capsule is presented. Histologically, apart from hyaline cartilage resembling chondroma the areas simulating chondrosarcoma were found. The differential diagnosis between synovial chondromatosis and chondrosarcoma is proposed. PMID- 2983646 TI - [Structural and functional bases of the mechanisms of myocardial fibrillation in sudden cardiac death]. AB - Analysis of current data indicates that one of the important and unsolved problems in the concept of sudden cardiac death is the question on the trigger mechanisms of the ventricular fibrillation which is the most frequent cause of heart failure in the sudden death of patients with coronary heart disease. It can be assumed that the spread, with the circulating blood, in the myocardial tissue of the arythmogenic substances formed in the hypoxic areas of the ischemized heart muscle or as an effect of catecholamines favour the fibrillation. Penetration of the arythmogenic substances into the circulation is due to the blood circulation remaining in the ischemic zone or the reperfusion of the ischemic tissue. Amphiphilic lysophosphoglycerides, cAMP, free fatty acids, lipid peroxidation products, some forms of prostaglandins may play a role of arythmogenic substances. PMID- 2983647 TI - Angiotensin-converting enzyme activity in human aqueous humor. AB - We measured aqueous angiotensin-converting enzyme (ACE) activity in 37 patients. Patients with granulomatous uveitis and probable sarcoidosis (based on increased serum ACE activity or characteristic radiologic findings) had a significant increase in aqueous ACE activity compared with normal subjects. Aqueous ACE activity was also significantly elevated in patients with sarcoid who had normal serum ACE activity. We believe that measurement of aqueous ACE activity should be considered in patients with uveitis who have normal serum ACE activity and are suspected of having sarcoidosis. PMID- 2983648 TI - Farber's disease. Light and electron microscopic study of the eye. AB - A 35-month-old girl had Farber's disease (disseminated lipogranulomatosis) manifested clinically by macular cherry-red spots. The pathologic changes consisted of intracellular inclusions of varying morphologic features and density. The most frequently encountered inclusion was 1.2 micron wide and consisted of flattened stacks of osmophilic lamellae (2.1 to 2.3 nm thick, with 4.4-nm periodicity) oriented in parallel or oblique array ("zebra-body" configuration) and enclosed by a focally discontinuous unit membrane. Some of the inclusions contained curved tubular profiles resembling curvilinear tubular bodies. The retinal ganglion cells were grossly distended with inclusions and showed the greatest pathologic changes. PMID- 2983649 TI - Sebaceous gland carcinoma. PMID- 2983650 TI - Interferon therapy for bronchial papillomatosis controlled by papillomavirus-DNA hybridization. AB - In biopsy specimens from a patient with tracheal and bronchial papillomatosis, human papillomavirus (HPV) type 11 DNA was identified. Treatment with leukocyte interferon was initiated, and the results of therapy was monitored by molecular hybridization of biopsy specimens with phosphorus 32-labeled HPV type 11 DNA after interferon application. There was no improvement of the clinical course, although two of five biopsy specimens showed a considerable reduction in the amount of HPV type 11. We discuss the reasons for the insufficient effect of therapy and propose molecular hybridization as an additional method for therapy control in viral diseases. PMID- 2983651 TI - Pleomorphic adenoma and metastatic squamous cell carcinoma. PMID- 2983652 TI - Prognostic determination and submandibular function in Bell's palsy. Dynamic testing with technetium Tc 99m. AB - In 56 patients with Bell's palsy, a submandibular gland function test employing a dynamic technetium Tc 99m scan was performed, to study the relationship between the test results and the prognosis of paralysis. Patients who had no difference in function on the affected facial side compared with the healthy side during the test (performed within ten days after symptomatic onset) had a high rate of early improvement. Patients who showed no response to the salivary secretory stimulant administered had an incomplete recovery and were all considered for surgery. Prognosis within ten days after onset was possible in as many as 37 of the 56 cases. PMID- 2983653 TI - Synovial sarcomas of the tongue base. The seventh reported case. AB - To our knowledge, we report the seventh synovial sarcoma of the base of the tongue in the world medical literature. Synovial sarcomas in this location are generally believed to arise from mesenchymal tissue above the hyoid. The surgical and radiation therapy of the primary and various chemotherapeutic programs used in the attempt to control the metastases are outlined. Early surgical therapy followed by radiation therapy before metastases is the best hope for cure. PMID- 2983654 TI - Exposure and direct stimulation of the semicircular canal cupula. AB - The ampullary wall of the posterior semicircular canal was isolated from the frog, cut and the cupula exposed in frog Ringer's solution. The cupula was stimulated by depression with a fine glass micropipette. Three points on the cupula were selected for depression: point A, the lowest point on the midline of the cupula surface on the utricular side; point B, the uppermost on the midline of the same surface; and point C, in the middle of the lateral surface of the cupula. Depression at points A and B was toward the canal, whereas depression at point C was toward the centre of the cupula. The amount of depression was controlled by a micromanipulator. At point A, the minimum depression facilitating the posterior canal action potential was 1 micron; at point B, a greater amount of depression was needed to produce the action potential. Even a large amount of depression at point C resulted in only a small action potential, possibly because the direction of the sensory cell polarity is along the long axis of the canal. PMID- 2983656 TI - Perinatal factors, periventricular haemorrhage and mortality in very low birthweight infants. AB - In a population of 225 very low birthweight infants born over a 21 month period the cerebroventricular system was scanned by ultrasound. One third of the infants developed a periventricular haemorrhage; in 41% of infants the haemorrhage was detected before an hour of age and 66% of all haemorrhages occurred within the first 24 hours. Statistically significant associations with periventricular haemorrhage included vaginal delivery, endotracheal intubation and intravenous sodium bicarbonate when this was administered in the first 24 hours. In a stepwise regression analysis, however, these and other potentially significant variables added little to the total accountable variance. A similar analysis of perinatal factors and mortality revealed that decreasing gestation was the major association with death. PMID- 2983655 TI - Freeze-fracture studies on the perilymph-endolymph barrier in experimentally induced hydrops. AB - A double-blind study was performed on nine pigmented guinea pigs which were unilaterally operated on in order to create an endolymphatic hydrops. After 12-18 months, both inner ears of the animals were removed and investigated by the freeze-fracture technique. The ultrastructure of the epithelial cells lining the endolymphatic compartment of the cochlea and the vestibular apparatus was analyzed. Special attention was directed to possible changes of the tight junctions (zonulae occludentes) in these sites. The number and depth of strands between the cells of sensory and non-sensory epithelia of both operated and unoperated sides were compared by an examiner unaware of the operated side. After the code of double-blind study was broken, it was found that there were no significant differences between the operated and the unoperated ears. These results are discussed in relation to the late DC-potential decrease in experimentally induced hydrops. PMID- 2983657 TI - [Leishmaniasis in dogs. Clinical course of infection, diagnosis and therapy trials following experimental infection of beagles with Leishmania donovani (Calcutta strain)]. PMID- 2983658 TI - Primary structure of the A chain of human complement-classical-pathway enzyme C1r. N-terminal sequences and alignment of autolytic fragments and CNBr-cleavage peptides. AB - Activated human complement-classical-pathway enzyme C1r has previously been shown to undergo autolytic cleavages occurring in the A chain [Arlaud, Villiers, Chesne & Colomb (1980) Biochim. Biophys. Acta 616, 116-129]. Chemical analysis of the autolytic products confirms that the A chain undergoes two major cleavages, generating three fragments, which have now been isolated and characterized. The N terminal alpha fragment (approx. 210 residues long) has a blocked N-terminus, as does the whole A chain, whereas N-terminal sequences of fragments beta and gamma (approx. 66 and 176 residues long respectively) do not, and their N-terminal sequences were determined. Fragments alpha, beta and gamma, which are not interconnected by disulphide bridges, are located in this order within C1r A chain. Fragment gamma is disulphide-linked to the B chain of C1r, which is C terminal in the single polypeptide chain of precursor C1r. CNBr cleavage of C1r A chain yields seven major peptides, CN1b, CN4a, CN2a, CN1a, CN3, CN4b and CN2b, which were positioned in that order, on the basis of N-terminal sequences of the methionine-containing peptides generated from tryptic cleavage of the succinylated (3-carboxypropionylated) C1r A chain. About 60% of the sequence of C1r A chain (440-460 residues long) was determined, including the complete sequence of the C-terminal 95 residues. This region shows homology with the corresponding parts of plasminogen and chymotrypsinogen and, more surprisingly, with the alpha 1 chain of human haptoglobin 1-1, a serine proteinase homologue. PMID- 2983659 TI - Evidence that the peripheral cyclic AMP phosphodiesterase of rat liver plasma membranes is a metalloenzyme. AB - Cyclic nucleotide phosphodiesterase activity in salt extracts of rat liver plasma membranes was progressively inactivated by treatment with the metal chelators 8 hydroxyquinoline and o-phenanthroline, but not the non-chelating m-phenanthroline isomer. Activity at 20 microM-cyclic AMP was lost more slowly than activity at 0.4 microM-cyclic AMP. The activity of treated preparations was partially restored by incubation with Zn2+ or Mn2+ ions (in the presence of 1 mM-MgCl2) but not with Ca2+, Cd2+, Co2+, Cu2+ or Fe2+ ions, nor by MgCl2 alone. The results suggest the presence in the membrane extracts of a cyclic AMP phosphodiesterase containing tightly bound metal, possibly Zn or Mn, that affects the enzyme's affinity for cyclic AMP. PMID- 2983660 TI - The crystal structure of beta-lactamase from Staphylococcus aureus at 0.5 nm resolution. AB - The preparation, crystallization and low-resolution structure determination of beta-lactamase (EC 3.5.2.6, 'penicillinase') from Staphylococcus aureus is described. The enzyme crystallizes in space group I222 with 1 molecule per asymmetric unit and cell dimensions a = 5.45(1), b = 9.39(1) and c = 13.87(2) nm. The structure was determined at 0.5 nm resolution by using phases calculated from (NH4)2Pt(CN)4 and KAu(CN)2 derivatives. The mean figure of merit mean value of m, for the 1106 reflexions used was 0.70. Difference Fourier syntheses for data collected from crystals soaked in platinum D-methionine and in 6-(4-hydroxy-3,5 di-iodobenzamido)penicilloic acid revealed the likely position of the active site of the enzyme. PMID- 2983661 TI - Guinea-pig liver testosterone 17 beta-dehydrogenase (NADP+) and aldehyde reductase exhibit benzene dihydrodiol dehydrogenase activity. AB - We have kinetically and immunologically demonstrated that testosterone 17 beta dehydrogenase (NADP+) isoenzymes (EC 1.1.1.64) and aldehyde reductase (EC 1.1.1.2) from guinea-pig liver catalyse the oxidation of benzene dihydrodiol (trans-1,2-dihydroxycyclohexa-3,5-diene) to catechol. One isoenzyme of testosterone 17 beta-dehydrogenase, which has specificity for 5 beta-androstanes, oxidized benzene dihydrodiol at a 3-fold higher rate than 5 beta dihydrotestosterone, and showed a more than 4-fold higher affinity for benzene dihydrodiol and Vmax. value than did another isoenzyme, which exhibits specificity for 5 alpha-androstanes, and aldehyde reductase. Immunoprecipitation of guinea-pig liver cytosol with antisera against the testosterone 17 beta dehydrogenase isoenzymes and aldehyde reductase indicated that most of the benzene dihydrodiol dehydrogenase activity in the tissue is due to testosterone 17 beta-dehydrogenase. PMID- 2983662 TI - Complete inhibition of dihydro-orotate oxidation and superoxide production by 1,1,1-trifluoro-3-thenoylacetone in rat liver mitochondria. AB - 1,1,1-Trifluoro-3-thenoylacetone was shown to cause complete inhibition of dihydroorotate oxidation in rat liver mitochondria as measured by orotate formation and the rate of dihydro-orotate-dependent reduction of 2,6 dichlorophenol-indophenol or cytochrome c. The inhibition by trifluorothenoylacetone was dose-dependent, and a concentration of 1 mM completely inhibited dihydro-orotate dehydrogenase activity. 1,10-Phenanthroline, another iron-chelating agent, also caused total inhibition of the liver enzyme. Whereas the iron chelators inhibited 100% of dihydro-orotate dehydrogenase activity in liver mitochondria, they inhibited only a maximum of 72% in the case of the brain enzyme. The inhibition by trifluorothenoylacetone was not prevented by addition of phenazine methosulphate or ubiquinone. Dihydro-orotate dehydrogenase-mediated generation of superoxide was abolished when the enzyme was fully inhibited by trifluorothenoylacetone or when the electron-transport system was blocked by antimycin A. These results suggest that the iron component(s) of dihydro-orotate dehydrogenase is of strategic importance for catalytic activity and transfer of reducing equivalents from the primary enzyme to the electron transport chain. Furthermore, the study indicates that production of superoxide radicals during dihydro-orotate dehydrogenase-catalysed oxidation of dihydro orotate may be at the cytochrome b-c1 segment of the electron-transport chain (as a consequence of autooxidation of ubisemiquinone) rather than at a site on the primary enzyme. PMID- 2983663 TI - Interspecies conservation of retinal guanosine 5'-triphosphatase. Characterization by photoaffinity labelling and tryptic-peptide mapping. AB - Light-activated hydrolysis of cyclic GMP is achieved through the photoexcitation of rhodopsin, a process which then triggers the replacement of GDP for GTP by a retinal guanosine 5'-triphosphatase referred to as 'transducin'. The transducin GTP complex then switches on the phosphodiesterase [Fung, Hurley & Stryer (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 152-156]. The bovine transducin consists of an alpha-subunit (39000 Mr), which is a GTP-binding component, together with a beta (37000 Mr) and a gamma-subunit (10000 Mr). We have purified retinal transducin from cow, pig, chick and frog. The enzyme specific activities and sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic profiles indicate that this enzyme is similar in all species except the frog. Whereas the bovine, pig and chick transducins consist of major 37000- and 39000-Mr components, that of the frog consists of a single 75000-Mr component. Labelling of the GTP-binding components with the photoaffinity label 8-azidoguanosine [gamma-32P]triphosphate demonstrated that the 37000-Mr components of the cow, pig and chick and the 75000 Mr component of the frog were major GTP-binding components. In addition, peptide maps of radioiodinated tryptic peptides indicate that the frog 75000-Mr protein is highly related to the pig transducin. These results demonstrate evolutionary conservation of retinal transducin and the presence of a higher-Mr, but nonetheless highly conserved form, of transducin in the frog. The relationship of this component to the recently reported rod-outer-segment inhibitor protein [Yamazaki, Stein, Chernoff & Bitensky (1983) J. Biol. Chem. 258, 8188-8194] is discussed. PMID- 2983665 TI - Functional receptors for epidermal growth factor in an epithelial-cell line derived from the rat small intestine. AB - Epidermal growth factor (EGF) regulates the proliferation of cells of a rat intestinal epithelial-cell line (RIE-1) in culture. Confluent RIE-1 cells were stimulated to proliferate by EGF with a half-maximal effect at 1-2 ng/ml. In contrast, the growth of sparse RIE-1 cells was inhibited by the growth factor. Binding studies at 4 degrees C with 125I-EGF identified two classes of binding sites for EGF on RIE-1 cells, one of high affinity (KD = 1.8 X 10(-10)M; 1.8 X 10(4) receptors/cell) and one of lower affinity (KD = 5.2 X 10(-9)M; 6.3 X 10(4) receptors/cell). After binding to the cells at 37 degrees C, 125I-EGF was rapidly internalized and subsequently degraded. Degradation products were released into the medium after a lag of 15-30 min. The degradation of 125I-EGF did not occur at 4 degrees C and was inhibited at 37 degrees C by chloroquine, methylamine or NH4Cl, but not by colchicine. Exposure of RIE-1 cells to EGF caused a time- and dose-dependent loss of EGF receptors from the cell surface. The recovery of receptors after the removal of EGF was retarded in the absence of serum and prevented by the presence of cycloheximide or actinomycin D. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis separation of the 125I-EGF-receptor complex from RIE-1 cells after covalently cross-linking with disuccinimidyl suberate indicated a receptor of Mr congruent to 160 000. The demonstration of functional EGF receptors in this cell line provides further evidence that EGF may regulate intestinal-epithelial-cell physiology. PMID- 2983664 TI - Hepatic endosome fractions contain an ATP-driven proton pump. AB - Endosome fractions were isolated from rat liver homogenates on the basis of the subcellular distribution of circulating ligands, e.g. 125I-asialotransferrin internalized by hepatocytes by a receptor-mediated process. The distribution of endocytosed 125I-asialotransferrin 1-2 min and 15 min after uptake by liver and a monensin-activated Mg2+-dependent ATPase activity coincided on linear gradients of sucrose and Nycodenz. The monensin-activated Mg2+-ATPase was enriched relative to the liver homogenates up to 60-fold in specific activity in the endosome fractions. Contamination of the endosome fractions by lysosomes, endoplasmic reticulum, mitochondria, plasma membranes and Golgi-apparatus components was low. By use of 9-aminoacridine, a probe for pH gradients, the endosome vesicles were shown to acidify on addition of ATP. Acidification was reversed by addition of monensin. The results indicate that endosome fractions contain an ATP-driven proton pump. The ionophore-activated Mg2+-ATPase in combination with the presence of undegraded ligands in the endosome fractions emerge as linked markers for this new subcellular organelle. PMID- 2983666 TI - Receptor-binding and down-regulatory properties of 22000-Mr human growth hormone and its natural 20000-Mr variant on IM-9 human lymphocytes. AB - Our earlier binding studies of the 22000- and 20000-Mr variants of human growth hormone (somatotropin) to pregnant-rabbit liver and mammary receptors [Closset, Smal, Gomez & Hennen (1983) Biochem. J. 214, 885-892] suggested that the 20000-Mr variant was a lower-affinity analogue of the 22000-Mr molecule. Since the receptor population in these tissues is not fully characterized, we have now investigated the binding of both variants to the well-characterized and highly specific human-growth-hormone receptor of the human lymphocyte IM-9 cell line. The maximum bindability of radioiodinated 22000- and 22000-Mr to IM-9 cells was 60 and 45% respectively. Both hormone variants have essentially the same binding characteristics: slow association (equilibrium reached in 8-10h at 30 degrees C), poor reversibility ('tight binding'), linear Scatchard plot, same specificity as shown by lack of competition by bovine, porcine or equine growth hormones or human growth hormone-(32-46)-(missing in the 20000-Mr variant),-(1-134)- and (141-191)-peptides. Both unlabelled hormones inhibit binding of both tracers completely, with the 20000-Mr variant being only half as potent as the 22000-Mr one. The apparent affinity is 2.8 X 10(9)M-1 for the 22000-Mr variant and 1.6 X 10(9)M-1 for the 20000-Mr variant. This decreased affinity of the 20000-Mr variant appears to be due to a lower association rate constant. Concentrations (5 ng/ml) of the two variants that occupy about 15% of the total sites induce a marked down-regulation of the receptors after 18h incubation, but the 20000-Mr variant (50% decrease) has a smaller effect than the 22000-Mr variant (75% decrease). Thus the only consequence of the residues-32-46 deletion in the 20000 Mr variant is a lower association rate and affinity for the IM-9 lymphocyte human growth-hormone receptor. The close binding characteristics of the two forms suggest that the known differences in their insulin-like effects cannot be explained by differences in the nature of their interaction with the human-growth hormone receptor. PMID- 2983667 TI - Histamine H2 receptors on chondrocytes derived from human, canine and bovine articular cartilage. AB - Histamine (1-100 microM) induced a concentration-dependent increase in intracellular cyclic AMP in monolayer cultures of human, canine and foetal-bovine articular chondrocytes. The dose-response curve for histamine in each culture was progressively displaced to the right with increasing concentrations of cimetidine, an H2-receptor antagonist. The histamine-induced cyclic AMP elevation in human articular chondrocytes was also significantly decreased by ranitidine, another H2 antagonist, but not by the H1 antagonists mepyramine and chlorpheniramine. These findings indicate that histamine activates chondrocyte adenylate cyclase through an H2 receptor. The cyclic AMP response of human chondrocytes to histamine was many times greater than that measured for synovial fibroblasts under similar conditions. Such findings suggest that mast-cell chondrocyte interactions in vivo may contribute to changed chondrocyte metabolism in joint disease. PMID- 2983668 TI - A 17 beta-hydroxysteroid dehydrogenase of female rabbit liver cytosol. Purification and characterization of multiple forms of the enzyme. AB - Multiple forms of the soluble 17 beta-hydroxysteroid dehydrogenase of female rabbit liver were identified. NAD-dependent and NADP-dependent enzyme activities were separated by affinity chromatography on agarose-immobilized Procion Red HE3B, and three forms of the NADP-dependent enzyme activity were purified by chromatofocusing. These three enzyme forms are charge isomers and have no quaternary structure. The enzymes catalysed the C-17 oxidoreduction of oestrogens and androgens; with all enzyme forms the activity towards androgens was higher than that toward oestrogens. The enzymes also exhibited 3 alpha-hydroxysteroid dehydrogenase activity towards androgens of the 5 beta-androstane series. Comparison of the relative activities of the enzymes towards a number of oestrogen and androgen substrates revealed differences among the enzyme forms for both the oxidative and the reductive reactions. In particular, one enzyme form had a significantly lower Km for the 3 alpha-hydroxysteroid substrate and a higher 3 alpha-/17 beta-hydroxysteroid dehydrogenase activity ratio than the other two enzyme forms. PMID- 2983669 TI - Age-dependent changes in the multiple forms of the soluble 17 beta-hydroxysteroid dehydrogenase of female rabbit liver. AB - The soluble NADP-dependent 17 beta-hydroxysteroid dehydrogenase activity of female rabbit liver increases with the age of the animal, the specific activity of the enzyme in the 56-day-old rabbit being 3 times that of the 28-day-old animal. The increase in activity is accompanied by a change in the molecular heterogeneity of the enzyme. Three forms (enzymes I, II and III) were identified in the liver cytosol of the 56-day-old female rabbit, whereas only one major form (enzyme IIIY) was present in the 28-day-old animal. Peptide maps of the four purified enzymes showed that there were minor differences in structure. The enzyme present in the liver of the 28-day-old rabbit was distinct from the three enzymes of the 56-day-old animal. All of the enzymes exhibited bifunctional activity, having 17 beta-hydroxysteroid dehydrogenase activity towards androgen and oestrogen substrates and 3 alpha-hydroxysteroid dehydrogenase activity towards androgens of the 5 beta-androstane series. The differences in substrate specificity of the enzymes paralleled their differences in structure. The data suggest that one enzyme (enzyme III) may have a special role in steroid metabolism during development in the female rabbit. PMID- 2983671 TI - Thermodynamic and steady-state-kinetic investigation of the effect of NN' dicyclohexylcarbodi-imide on H+ translocation by the mitochondrial cytochrome bc1 complex. AB - Steady-state kinetic measurements showed that NN'-dicyclohexylcarbodi-imide decreased the observed H+/2e ratio of H+ transport by mitochondria respiring on succinate, acting mainly at the cytochrome bc1 complex. Thermodynamic assessment of the H+/2e ratio by measuring the force ratio across the bc1 complex showed that the inhibitor did not affect H+ translocation. Possible explanations of this disagreement between methods are examined; we conclude that the inhibitor does not alter the mechanistic stoichiometry of H+ pumping by the bc1 complex. PMID- 2983670 TI - Thermodynamic control of electron flux through mitochondrial cytochrome bc1 complex. AB - The redox states of exogenously added ubiquinone-2 and cytochrome c, and the protonmotive force (delta p) of rat liver mitochondria were measured as the respiration rate was titrated with the uncoupler carbonyl cyanide p trifluoromethoxyphenyl-hydrazone. The force ratio delta Eh/delta p across the bc1 complex was close to 1:1 in State 4, indicating an H+/e- stoichiometry of 1:1 for the cytochrome bc1 complex, excluding protons moved by pool ubiquinone. Assuming a constant stoichiometry the rate of electron transport increased linearly with the disequilibrium (delta Eh - delta p) across the complex. PMID- 2983672 TI - Electroneutral efflux of Ca2+ from liver mitochondria. AB - Respiring liver mitochondria were allowed to export Ca2+ on the endogenous Ca2+/nH+ antiporter in the presence of Ruthenium Red (to inhibit uptake on the Ca2+ uniporter) until a steady state was reached. Addition of sufficient of the ionophore A23187 (which catalyses Ca2+/2H+ exchange) to bring the Ca2+ and H+ gradients into equilibrium did not alter the steady state. Thermodynamic analysis showed that if a Ca2+/nH+ exchange with any value of n other than 2 was at equilibrium, addition of A23187 would have caused an easily measurable change in extramitochondrial free [Ca2+]. Therefore, the endogenous carrier of liver mitochondria catalyses electroneutral Ca2+/2H+ antiport. PMID- 2983673 TI - Regulation of fructose 2,6-bisphosphate concentration in white adipose tissue. AB - Injection of insulin to fed rats diminished the concentration of fructose 2,6 bisphosphate in white adipose tissue. Incubation of epididymal fat-pads or adipocytes with insulin stimulated lactate release and sugar detritiation and also decreased fructose 2,6-bisphosphate concentration. Such a decrease was, however, not observed in fat-pads from starved or alloxan-diabetic rats. Incubation of adipocytes from fed rats with various concentrations of glucose or fructose led to a dose-dependent rise in fructose 2,6-bisphosphate which correlated with lactate output and detritiation of 3-3H-labelled sugar. In adipocytes from fed rats, palmitate stimulated the detritiation of [3-3H]glucose without affecting lactate production and fructose 2,6-bisphosphate concentration. Incubation of epididymal fat-pads from fed rats in the presence of antimycin stimulated lactate output but decreased fructose 2,6-bisphosphate concentration. Changes in lipolytic rates brought about by noradrenaline, insulin, adenosine and corticotropin in adipocytes from fed rats were not related to changes in fructose 2,6-bisphosphate or to rates of lactate output. In fed rats, the activity of 6 phosphofructo-2-kinase was not changed after treatment of adipocytes with insulin, noradrenaline or adenosine. It is suggested that the decrease in fructose 2,6-bisphosphate concentration observed after insulin treatment can be explained by the increase in sn-glycerol 3-phosphate, an inhibitor of 6 phosphofructo-2-kinase. PMID- 2983674 TI - ADP-ribosyltransferase in isolated nuclei from sea-urchin embryos. AB - The activity of ADP-ribosyltransferase in nuclei isolated from sea-urchin embryos was estimated by the incorporation of [adenosine-14C]NAD+ into the acid-insoluble fraction. Hydrolysis of this acid-insoluble product by snake venom phosphodiesterase yielded radioactive 5'-AMP and phosphoribosyl-AMP. The incorporation of [14C]-NAD+ was inhibited by 3-aminobenzamide and nicotinamide, potent inhibitors of ADP-ribosyltransferase. [14C]NAD+ incorporation into the acid-insoluble fraction results from the reaction of ADP-ribosyltransferase. The optimum pH for the enzyme in isolated nuclei was 7.5. The enzyme, in 50 mM Tris/HCl buffer, pH 7.5, containing 0.5 mM-NAD+ and 0.5 mM-dithiothreitol, exhibited the highest activity at 18 degrees C in the presence of 14 mM-MgCl2. The apparent Km value for NAD+ was 25 microM. The activity of the enzyme was measured in nuclei isolated from the embryos at several stages during early development. The activity was maximum at the 16-32-cell stage and then decreased to a minimum at the mesenchyme blastula stage. Thereafter its activity slightly increased at the onset of gastrulation and decreased again at the prism stage. PMID- 2983675 TI - Inhibition of class C beta-lactamases by (1'R,6R)-6-(1' hydroxy)benzylpenicillanic acid SS-dioxide. AB - beta-Lactamases, enzymes that catalyse the hydrolysis of the beta-lactam ring in beta-lactam antibiotics, are divided into three classes, A, B and C, on the basis of the structures so far determined. There are relatively few effective inhibitors of class C beta-lactamases. A beta-lactam sulphone with a hydroxybenzyl side chain, namely (1'R,6R)-6-(1'-hydroxy)benzylpenicillanic acid SS-dioxide (I), has now been studied. The sulphone is a good mechanism-based inhibitor of class C beta-lactamases. At pH8, the inhibition of a Pseudomonas beta-lactamase is irreversible, and proceeds at a rate that is about one-tenth the rate of concurrent hydrolysis. The labelled enzyme has enhanced u.v. absorption and is probably an enamine. At a lower pH, however, inhibition is transitory. PMID- 2983676 TI - The methane mono-oxygenase reaction system studied in vivo by membrane-inlet mass spectrometry. AB - A membrane-inlet mass spectrometer connected to an open-system cuvette was used for direct measurement of dissolved methane and O2 in bacterial samples of strain OU-4-1, a type II methanotrophic bacterium. A technique was applied for keeping the concentration of dissolved methane or O2 in the sample constant while the concentration of the other dissolved gas was varied. This allowed the reaction mechanism of methane mono-oxygenase to be studied in vivo. The enzyme was found to follow a random bi-reactant mechanism with respect to binding of methane and O2. Binding of one substrate decreased the affinity for the other. The true binding constants were 1 microM for methane and 0.14 microM for O2. Studies of HCN inhibition confirmed the random bi-reactant mechanism. HCN was found to be a non-exclusive inhibitor with a binding constant of 0.4 microM. PMID- 2983677 TI - Enhancement of DNA-mediated gene transfer by high-Mr carrier DNA in synchronized CV-1 cells. AB - Synchronized CV-1 cells were transfected with SV40 (simian virus 40) DNA-calcium phosphate co-precipitates. In the presence of carrier DNA, the transfection efficiency of SV40 DNA was decreased 5-fold in S-phase cells and was increased 4 fold in preparations of mitotically enriched cells as compared with asynchronous controls. No difference was observed when carrier DNA was omitted, when cells had progressed through S-phase and into G2-phase, or when the infectivity of cells to intact SV40 virus was tested. These results highlight the importance of cell cycle-dependent factors on DNA-mediated gene transfer. PMID- 2983678 TI - The lanthanide-enhanced affinity chromatography of clostridial collagenase. AB - Clostridiopeptidase A (EC 3.4.24.3) did not bind to a collagen affinity column in the absence of Ca2+, but did so in the presence of lanthanide ions (Ln3+). The sequestered enzyme could be eluted with EGTA. For the four Ln3+ ions tested, the order of efficiency in promoting enzyme binding, Sm3+ greater than Lu3+ greater than Er3+ much greater than La3+, reflected their relative abilities to inhibit clostridiopeptidase A. By using Sm3+ as an adjunct, it proved possible to separate a highly active preparation of collagenase from crude clostridial collagenase. Sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic analysis of the preparation revealed a major protein of Mr 95000 and a minor component of Mr 82000. As both were stained by periodic acid/Schiff reagent, they were probably glycoproteins. PMID- 2983680 TI - Contribution of different protein phosphatases to the dephosphorylation of 6 phosphofructo-1-kinase and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase in rat liver. AB - The nature of rat liver protein phosphatases involved in the dephosphorylation of the glycolytic key enzyme 6-phosphofructo-1-kinase and the regulatory enzyme 6 phosphofructo-2-kinase/fructose 2,6-bisphosphatase was investigated. In terms of the classification system proposed by Ingebritsen & Cohen [(1983) Eur. J. Biochem. 132, 255-261], only the type-2 protein phosphatases 2A (which can be separated into 2A1 and 2A2) and 2C act on these substrates. Fractionation of rat liver extracts by anion-exchange chromatography and gel filtration revealed that protein phosphatase 2A is responsible for most of the 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase phosphatase activity (activity ratio 2A/2C = 4:1). On the other hand, 6-phosphofructo-1-kinase phosphatase activity is equally distributed between protein phosphatases 2A (2A1 plus 2A2) and 2C. In addition, the possible role of low-Mr compounds for the control of purified protein phosphatase 2C was examined. At near-physiological concentrations, none of the metabolites studied significantly affected the rate of dephosphorylation of 6 phosphofructo-1-kinase, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase, pyruvate kinase or fructose-1,6-bisphosphatase. PMID- 2983679 TI - Extracellular-matrix synthesis by skeletal muscle in culture. Major secreted collagenous proteins of clonal myoblasts. AB - We have previously shown that G8-1, a murine clonal skeletal-muscle cell line, produces a substrate-attached extracellular matrix [Beach, Burton, Hendricks & Festoff (1982) J. Biol. Chem. 257, 11437-11442]. To examine further the expression of extracellular-matrix proteins by muscle cells, we have analysed the collagenous proteins secreted by G8-1 myoblasts. We have found that collagens and/or procollagens, corresponding to genetic types I, III and IV (and possibly V), are produced and secreted by G8-1 myoblasts. The major secreted collagenous polypeptides were identified as alpha 1 type I and its precursors by using pulse chase studies, pepsin and collagenase digestions and CNBr fragmentation. The presence of lesser amounts of the other collagens was determined by immunoprecipitation. These results demonstrate that clonal skeletal-muscle cells, in the absence of fibroblasts and an exogenous collagen substrate, are able to synthesize and secrete several extracellular-matrix collagenous proteins in proportions similar to those which are commonly found in muscle tissue and mixed cultures of muscle cells and fibroblasts. PMID- 2983681 TI - Inhibition of hyaluronate synthesis. AB - UDP-GlcNAc (UDP-N-acetylglucosamine) and UDP-GlcA (UDP-glucuronic acid) were oxidized by periodate in the ribose ring and utilized as inhibitors for hyaluronate synthase in membrane fractions from the B6 cell line and in cell cultures of B6 cells and human fibrosarcoma HT 1080. Inhibition was irreversible and concentration-dependent and could be prevented in the case of periodate oxidized UDP-GlcNAc by UDP-GlcNAc. Periodate-oxidized UDP-GlcNAc was shown to block chain elongation of hyaluronate. Introduction of periodate-oxidized UDP GlcA into B6 cells by hypo-osmotic lysis of pinocytotic vesicles decreased hyaluronate synthesis by direct inhibition of the synthase. In HT 1080 cells the synthesis of hyaluronate, chondroitin sulphate and heparan sulphate was inhibited simultaneously. PMID- 2983682 TI - Affinity-labelling of the thyrotropin receptor. Characterization of the photoactive ligand. AB - Thyrotropin (TSH) has been coupled to the photoactive heterobifunctional reagent N-hydroxysuccinimidyl 4-azidobenzoate (HSAB) and the properties of the product (HSAB-TSH) investigated. Preparations of HSAB-TSH containing two molecules of HSAB per molecule of TSH were used in most experiments and these preparations retained about 40% of the original receptor-binding activity of the TSH. HSAB-TSH could be labelled with 125I and cross-linked to porcine and human TSH receptors. Analysis of the cross-linked complexes indicated that the receptors consisted of two subunits (designated A and B) linked by a disulphide bridge. In the case of the human TSH receptor, the A- and B-subunits had approximate Mr values of 50 000 and 30 000 respectively, whereas the Mr values for porcine TSH-receptor A- and B subunits were approx. 45 000 and 25 000 respectively. Only the A subunit was cross-linked to TSH. Comparison of the effects of trypsin and mercaptoethanol on the TSH-TSH-receptor complexes suggested that the trypsin cleavage point on the A subunit was at a point close to the disulphide bridge. PMID- 2983684 TI - The respiration and calcium content of heart mitochondria from rats with vitamin D-induced cardionecrosis. AB - Mitochondria were isolated from the heart and skeletal muscle of rats treated with three consecutive daily doses of 100 000 i.u. of calciol (cholecalciferol; 'vitamin D3'). On the fourth day after the last dose, cardiac necrosis developed. At that time mitochondria isolated from heart displayed a 10-fold higher Ca2+ content and a 6-fold lower respiratory rate with pyruvate-plus-malate as substrate as well as with other NAD-dependent substrates. No decrease in respiratory rate with succinate as substrate was observed. EDTA (5 mM) added to the medium during the isolation procedure restored both the high respiratory rate with pyruvate + malate and the low Ca2+ content of the heart mitochondria. The addition of 1 mM-CaCl2 to the medium in which a healthy (control) rat heart had been homogenized caused the same impairment of the mitochondria as did calciol treatment of the animals. No changes of mitochondria isolated from skeletal muscle were observed in rats treated with calciol. It is concluded that the heart mitochondria in vivo fail to accumulate Ca2+ from the cardiac cell overloaded with Ca2+ as the consequence of calciol treatment. Mitochondrial Ca2+ accumulation occurs during the isolation procedure unless an appropriate amount of chelating agent is added to the homogenization medium. The implication of these findings for the biochemical sequence of events in the calciol-induced cardiac necrosis is discussed. PMID- 2983683 TI - Redistribution of membrane-bound and cytosolic action in rabbit polymorphonuclear leucocytes during phagocytosis. AB - In the analysis of highly purified surface membrane from both resting and phagocytosing neutrophils an increase in the surface membrane associated actin has been demonstrated. This change at the cell periphery is associated with a coincident increase in the F-actin content of the cells following stimulation of the cells by exposure to opsonized Oil Red O droplets. The actin which is newly associated with the surface membrane of the phagocytosing cells was more susceptible to removal by detergent than the membrane-associated actin in resting cells and it was also noted that the F-actin associated with phagosomes was readily disrupted by detergent. A redistribution of the surface membrane glycoprotein 5'-nucleotidase was observed during phagocytosis, but no change in distribution of a 125I-labelled Lens culinaris lectin was observed during the entire phagocytic process. PMID- 2983685 TI - The role of adenosine kinase in regulating adenosine concentration. PMID- 2983686 TI - Effect of thyroid state on cyclic AMP-mediated induction of hepatic phosphoenolpyruvate carboxykinase. AB - Hepatic phosphoenolpyruvate carboxykinase (PEPCK) is significantly increased in the hyperthyroid starved rat, and moderately decreased in the hypothyroid starved rat. As tri-iodothyronine by itself has only a small and sustained effect on the induction of this enzyme, as was previously shown in the isolated perfused organ, the effect of hypo- and hyper-thyroidism on the increase in cytosolic PEPCK provoked by dibutyryl cyclic AMP (Bt2cAMP) was investigated in vivo and in the isolated perfused liver. Compared with euthyroid fed controls, in hypothyroid fed rats Bt2cAMP provoked in 2 h only a small increase in translatable mRNA coding for PEPCK. In contrast, in hyperthyroid animals PEPCK mRNA as measured by translation in vitro was already increased in the fed state, and further enhanced by Bt2cAMP injection to values as in euthyroid controls. Under all thyroid states a close correlation between PEPCK mRNA activity and PEPCK synthesis was observed. In the isolated perfused liver from the hyperthyroid fed rat, the increase in PEPCK provoked by Bt2cAMP or Bt2cAMP + isobutylmethylxanthine was considerably enhanced compared with those obtained in livers of hypothyroid rats. Also, adrenaline provoked a stimulated induction of PEPCK in hyperthyroid rats compared with hypothyroid rats. To summarize, our data indicate that the primary action of thyroid hormones on the synthesis of hepatic cytosolic PEPCK is to accelerate the cyclic AMP- or adrenaline-induction of the enzyme, acting primarily at a pretranslational level. PMID- 2983687 TI - The isolation of nuclear envelopes. Effects of thiol-group oxidation and of calcium ions. AB - The effects of (a) oxidative cross-linking of protein thiol groups and (b) the presence or absence of Ca2+ ions on rat liver nuclear-envelope isolation were studied. Two envelope-isolation procedures were compared: a well characterized low-ionic-strength method and a recently developed high-ionic-strength method. The latter method seems preferable to the former in respect of lower intranuclear contamination of the envelopes, suppression of endogenous serine proteinase, and maintenance of high specific activities of envelope-associated enzymes. In both procedures, however, the presence of Ca2+ gave rise to a rapid, apparently irreversible, contamination of the envelopes by intranuclear material. This effect was half-maximal at 20 microM-Ca2+. In addition, the envelopes became contaminated with intranuclear material by a Ca2+-independent mechanism, apparently resulting from N-ethylmaleimide-sensitive intermolecular disulphide bond formation. This oxidative process seemed to have two major kinetic components (half-life, t1/2, approx. 2 min and 10 min). In view of these findings, it is recommended that (i) for most purposes, nuclear envelopes be isolated by the newly developed high-ionic-strength procedure, (ii) irrespective of the method used, Ca2+-chelators be included in all the buffers, (iii) thiol group oxidation be prevented or reversed during the procedure. PMID- 2983688 TI - Difluoromethylornithine-induced amplification of ornithine decarboxylase genes in Ehrlich ascites carcinoma cells. AB - Stepwise increments of the concentration of 2-difluoromethylornithine, a mechanism-based irreversible inhibitor of mammalian ornithine decarboxylase (EC 4.1.1.17), resulted in a selection of cultured Ehrlich ascites carcinoma cells capable of growing in the presence of up to 50 mM difluoromethylornithine. Dialyzed extracts of drug-resistant tumor cells exhibited a very high ornithine decarboxylase activity and contained large excess of immunoreactive ornithine decarboxylase protein. Hybridization analyses with cloned complementary DNA revealed that the difluoromethylornithine-resistant tumor cells also expressed mRNA of the enzyme at greatly enhanced rate. The overproduction of ornithine decarboxylase by the tumor cells grown under the pressure of difluoromethylornithine was at least partly attributable to a 10 to 20-fold increase in the total gene dosage of ornithine decarboxylase involving an amplification of several genes of the gene family. The gene amplification developed appeared to be stable, as the gene dosage only slowly (during a period of several months) returned towards the normal level upon the removal of difluoromethylornithine. The overproduction of ornithine decarboxylase was accompanied by an enhanced resistance of the enzyme towards difluoromethylornithine in vitro. PMID- 2983689 TI - Effects of dimethylsulfoxide on sphingomyelinase in cultured human fibroblasts and correction of sphingomyelinase deficiency in fibroblasts from Niemann-Pick patients. AB - The effects of dimethylsulfoxide (DMSO) on sphingomyelinase activity were studied using human skin fibroblasts from normal individuals and Niemann-Pick patients. Sphingomyelinase activity in normal fibroblasts increased up to 230% of controls by 2% DMSO while the cell growth was inhibited. Other lysosomal hydrolases showed a rather smaller extent of increment in activity. There was no direct effect of DMSO in cell-free system and no evidence of any activating factor of sphingomyelinase in homogenates. Sphingomyelinase deficiency in fibroblasts from a Niemann-Pick patient (type C) was corrected by 2% DMSO with regaining its enzyme activity. PMID- 2983690 TI - Coupled endo-exodeoxyribonuclease activity from HeLa cell nuclei interacts with specific regions of the SV40 mini chromosome. AB - A recently described endo-exodeoxyribonuclease activity (1) was determined to interact preferentially with the origin/enhancer region and the 5' end of the late promoter region of the SV40 mini chromosome. The enzymatic activity has no preferential site of interaction with naked DNA, which suggests, the chromatin organization is responsible for this site directed interaction. PMID- 2983691 TI - Coupled endo-exodeoxyribonuclease activity from HeLa cell nuclei can function in an excision repair mode in vitro. AB - A single strand specific coupled endo-exodeoxyribonuclease activity has been purified and characterized from the nuclei of HeLa cells(7). The novelity of this activity is in the nature of its associated endo and exonuclease activities. Due to this aspect the ability of this activity to work in unison with DNA polymerase molecules in the process of DNA repair synthesis was investigated. PMID- 2983692 TI - Interaction of calmodulin and other calcium-modulated proteins with mammalian and arthropod junctional membrane proteins. AB - Calmodulin and other calcium-modulated proteins bind in vitro to purified junctional polypeptides from rat liver gap junctions, bovine lens fiber junctions, a chymotryptic fragment from bovine lens junctions, and crayfish hepatopancreas gap junctions. The potential biological relevance of the interaction of calmodulin with junctional proteins is suggested by immunocytochemical localization of endogenous calmodulin in cortical regions of the cell where gap junctions exist. These observations provide a molecular basis for understanding the potential regulatory role of calmodulin on cell-cell communication channels in vivo. In addition, the calmodulin binding represents the first molecular homology that has been found for junctional channel proteins from mammalian and arthropod tissues. PMID- 2983693 TI - Spin-trapping of methyl radical in the oxidative metabolism of 1,2 dimethylhydrazine. AB - A carbon-centered free radical formed during oxidative metabolism of 1,2 dimethylhydrazine has been spin-trapped with alpha-(4-pyridyl-1-oxide)N-tert butyl nitrone and 2-methyl-2-nitrosopropane. In the horseradish peroxidase/H2O2 catalyzed oxidation, the trapped species was identified as the methyl radical by the characteristic 1:3:3:1 quartet pattern of the 2-methyl-2-nitroso propane adduct. A carbon-centered radical is also formed during microsomal oxidation of 1,2-dimethylhydrazine in the presence of NADPH. However, the alpha-(4-pyridyl-1 oxide)N-tert-butyl nitrone trapped radical has not been unambiguously identified in this latter instance. These results may be of importance in regard to both carcinogenic and antitumor properties of 1,2-disubstituted hydrazine derivatives. PMID- 2983695 TI - Spectroscopic studies on the reaction of superoxide ion with tocopherol model compound, 6-hydroxy-2,2,5,7,8-pentamethylchroman. AB - The reaction of superoxide ion, O2-, with alpha-tocopherol model compound, 6 hydroxy-2,2,5,7,8-pentamethylchroman (1b), was investigated spectrophotometrically in acetonitirle. The transient absorption (lambda max = 330 nm) observed at the initial stage of the reaction was ascribed to the chromanoxyl-type radical, one-elecron oxidation product of compound 1b, on the basis of the spectroscopic [ultraviolet (UV)/visible and electron spin resonance (ESR)] data. Further, the final product observed was ascribable to the tocopherol quinone (3). PMID- 2983694 TI - Nuclear ADP-ribosylation in the chick lens during embryonic development. AB - Nuclear ADP-ribosyltransferase is present in cells from the chick lens throughout embryonic development. The activity does not decrease when the cells become post mitotic and commence terminal differentiation but declines slowly in both epithelia and fibre cells. At all stages studied the enzyme retains its ability to be activated by DNA strand breaks induced either by X-irradiation or by the action of an endogenous endonuclease. There is no correlation between the enzyme activity or the levels of its substrate NAD+ and the changes in DNA repair capacity which have been observed during the development of the lens. PMID- 2983696 TI - Inhibitory action of phosphatidylinositol on synaptosomal (Na+ + K+)-ATPase activity. AB - Phosphatidylinositol and several other phospholipids were tested for their ability to influence the (Na+ + K+)-ATPase activity of the cortical synaptic membrane from rats at various levels of free Ca2+. Phosphatidylinositol, but not phosphatidylethanolamine, phosphatidylcholine nor phosphatidylserine, markedly inhibited this enzyme activity, when the free Ca2+ concentration in the incubation media was less than 2.5 X 10(-6) M. This result suggests that phosphatidylinositol may play a role in the depolarization and/or the release of neurotransmitters or intracellular substances in the brain. PMID- 2983697 TI - Cyclic AMP and the heat shock response in Chinese hamster ovary cells. AB - Heat shock leads to transient increases in cAMP levels in HA-1-CHO cells. Such pulses are correlated temporally with the induction of heat resistance (thermotolerance) and with heat shock protein synthesis. Although the kinetics of cAMP increase after heating suggest a role in thermotolerance induction, raising cAMP levels directly using dBcAMP did not produce full thermotolerance. The resistance induced by dBcAMP may thus be either a component of or different to heat-shock triggered resistance. Cells which had been made thermotolerant by heat shock did not produce a pulse in cAMP level on heating. The cAMP producing system thus seemed desensitized to heat in thermotolerant cells. PMID- 2983698 TI - Heme-copper relationship of cytochrome oxidase in rat brain in situ. AB - The role of copper aa3 in relation to heme aa3 of cytochrome oxidase in electron transfer and oxygen utilization is poorly understood in vitro. In an attempt to study this in situ, we have simultaneously monitored the steady state redox changes of heme aa3 and copper aa3 in an isolated perfused rat head model (skull intact, muscle removed). By means of reflectance spectrophotometry the redox reactions of heme aa3 and copper aa3 were continuously monitored using 605-625 nm and 815-920 nm wavelength pairs, respectively. The reaction kinetics of these components in response to transient perfusion interruption in energized and de energized preparations were then examined. We found that in response to perfusion interruption, soon after full reduction, the copper signal begins to change toward oxidation despite continuation of anoxic insult and progressive reduction of heme aa3. This phenomenon disappeared by pretreatment of the preparation with 2,4-dinitrophenol. A schematic sequence of electron transport in situ is proposed which emphasizes an active role for Cua in this sequence. PMID- 2983699 TI - Platelet membrane molecular organization: relationship with membrane bound calcium. AB - The fatty acid spin label 5 nitroxide stearate has been used to determine the membrane organization changes induced by platelet aggregation. A decrease in order is observed with thrombin, even in the presence of EDTA, when aggregation is inhibited. Conversely, after aggregation by the calcium ionophore A23187 the rigidity of the phospholipids is not modified. These effects are discussed in relation to the release of membrane bound calcium induced by thrombin. PMID- 2983700 TI - Phosphorylation of the band 4.1-like proteins of the bovine lens. AB - Crude membrane fractions prepared from the superficial cortical fiber cells of bovine lenses were incubated with [gamma-32P]-ATP. Membrane-associated 4.1-like proteins were then immunoprecipitated and analyzed for 32P-incorporation. Band 4.1-like proteins of 150 kd, 80 kd, and 78 kd were found to be labeled in a time dependent fashion, with the 150 kd protein incorporating label at an 8-fold greater rate than the 80 kd and 78 kd proteins. Addition of cAMP or Ca2+ to the membrane preparations stimulated the phosphorylation of all three of these proteins. Our results indicate that the 4.1-like proteins of the lens fiber cell share with band 4.1 of the RBC the characteristic of being substrates for cAMP independent and cAMP-dependent protein kinases. PMID- 2983702 TI - Inhibition of sarcoplasmic reticulum Ca2+-ATPase by 2-mercaptoethanol. AB - The Ca2+-dependent ATPase activity of sarcoplasmic reticulum was inhibited when membrane vesicles were incubated at 0 degree C in presence of thiols. 2 mercaptoethanol was the most effective inhibitor from the thiols tested. The effect of 2-mercaptoethanol on the ATPase activity was biphasic; enzyme inhibition originally increased and then decreased with increasing thiol concentration. The inhibitory action of this thiol was significantly higher at low membrane concentrations and the rate of inactivation at 22 degrees C was considerably lower than that at 0 degree C. Ca2+-ATPase previously inhibited by 2 mercaptoethanol was partially reactivated by incubation with periodate. PMID- 2983701 TI - Pertussis but not cholera toxin inhibits the stimulated increase in actin association with the cytoskeleton in rabbit neutrophils: role of the "G proteins" in stimulus-response coupling. AB - Treatment of rabbit neutrophils with pertussis toxin, but not cholera toxin, inhibits the increases produced by formylmethionyl-leucyl-phenylalanine, leukotriene B4 and the calcium ionophore A23187 in the amounts of actin associated with the cytoskeletons. The increase in the cytoskeletal actin produced by phorbol 12-myristate, 13-acetate on the other hand is not affected by pertussis toxin. Incubation of the neutrophils with cholera toxin, unlike pertussis toxin, did not inhibit the fMet-Leu-Phe induced rise in the intracellular concentration of free calcium, and caused only a shift to the right of the dose-response curve of N-acetyl-beta-glucosaminidase release. This shift was more marked in the presence of 1-methyl-3-isobutylxanthine. In addition, the stimulated breakdown of phosphatidylinositol 4,5 bis-phosphate was inhibited by pertussis toxin. These results suggest that pertussis toxin acts at an early step in the signal transduction and does not affect the sequence of reactions initiated by the activation of the protein kinase C. Furthermore, the guanine nucleotide regulatory protein Gi, but not Gs, is closely involved in signal transduction in these cells. PMID- 2983703 TI - Tumor promoter phorbol-12-myristate-13-acetate induces poly ADP-ribosylation in human monocytes. AB - The tumor promoter phorbol-12-myristate-13-acetate (PMA) induces rapid poly ADP ribosylation and a drop in cellular NAD concentration in human monocytes. The antioxidants CuZn-superoxide dismutase, catalase, glutathione peroxidase and butylated-hydroxytoluene inhibit the reaction indicating that active oxygen species produced in the PMA-induced oxidative burst represent intermediates. The inhibitor of ADP-ribosyl-transferase, 3-amino-benzamide, inhibited poly ADP ribosylation but did not prevent the drop in NAD-levels. PMA also causes the slow accumulation of DNA strand breaks in monocytes. The difference in the kinetics of poly ADP-ribosylation and DNA breakage argues against a simple relationship between the two reactions. PMID- 2983704 TI - Characterization of pyrophosphate exchange by the reconstituted adenine nucleotide translocator from mitochondria. AB - The transport of inorganic pyrophosphate (PPi) by the adenine nucleotide translocator from beef heart mitochondria was studied in a reconstituted system. The transport of PPi is dependent on appropriate transmembrane substrates. The activity of PPi exchange is about one tenth as compared to the ADP/ATP exchange, whereas the transport affinity for PPi is very low (2-5 mM). The adenine nucleotide carrier catalyzes a strict counterexchange of PPi and nucleotides with an exchange stoichiometry close to 1. The inhibitor specificity of PPi exchange is comparable to that of ADP/ATP exchange. PMID- 2983705 TI - Peritoneal macrophages of guinea pig possibly lack LTC4 synthetase. AB - Peritoneal cells and adherent cells of mice and rats synthesized LTC4 and LTB4 when stimulated with A23187 in vitro. On the other hand, neither peritoneal cells nor adherent cells of guinea pigs generated LTC4, D4, and E4, but did the lower amounts of LTB4. Only generation of LTB4 was potentiated by simultaneous addition of 10 microM A.A. in this species. Enzyme solutions which were extracted from peritoneal cells of these three species were capable of converting DNCB to a colored product in the presence of glutathione and then these potencies were in the following order; guinea pig greater than mouse greater than rat. On the other hand, the potencies of converting LTA4 to LTC4 in the presence of glutathione were in the following order; mouse greater than rat much greater than guinea pig approximately equal to 0. These results suggest that macrophages of guinea pigs lack "LTC4 synthetase" and also this enzyme is different from usual GSH S transferases. PMID- 2983706 TI - Purification and properties of the phosphate eliminating enzyme involved in the biosynthesis of BH4 in man. AB - An enzyme catalyzing the elimination of triphosphate from 7,8-dihydroneopterin triphosphate in the presence of Mg2+ has been purified approx. 3000 fold from human liver. It has a molecular weight of approx. 63'000, a pI value of 4.4 - 4.6 and is stable at 80 degrees C for 5 min. This enzyme catalyzes the formation of tetrahydrobiopterin in the presence of sepiapterin reductase, Mg2+ and NADPH. It is thus possible, that it also catalyzes the internal oxidoreduction leading to formation of the intermediate 6-pyruvoyl-tetrahydropterin, suggesting that no further enzyme is obligatory for biosynthesis of tetrahydrobiopterin. PMID- 2983707 TI - Estrogen synthetase stimulation by hemin in human choriocarcinoma cell culture. AB - The ability of hemin to stimulate estrogen synthetase (aromatase) in cultured human trophoblast cells and in cellular homogenates was investigated and compared with aromatase stimulation by dibutyryl cAMP [(Bu)2 cAMP]. Cells grown with hemin for 24 h, or homogenates incubated for 45 min with hemin, showed maximal aromatase stimulation (150 to 200% of activities in the absence of hemin) at 25 microM and 0.1 microM, respectively. Aromatase stimulation in culture by 25 microM hemin was observed within 4 h after hemin addition, while (Bu)2 cAMP required more than 6 h. Intracellular heme and porphyrin levels were higher (160 to 185%) in 96 h (Bu)2 cAMP-grown cells than control cells. PMID- 2983708 TI - Enhancer-facilitated expression of a human H4 histone gene. AB - Cultured mammalian cells were transfected with a recombinant human H4 histone gene. S1 nuclease mapping of cellular RNAs from transfected cells revealed: (i) correct initiation of transcription at the cap site, with some transcripts originating from other sites in the 5' flanking region of this H4 gene; (ii) cis linkage of an SV-40 transcriptional enhancer element upstream of the H4 5' flanking region resulted in about a 50-fold increase in the level of correctly initiated H4 mRNA and (iii) in a heterologous murine system stability of human H4 mRNAs was apparently sensitive to inhibition of DNA-synthesis by hydroxyurea. Our results suggest that certain sequences required for the initiation of a human H4 histone gene transcript reside within the 210 nucleotides immediately upstream from the cap site and that the level of expression is influenced by the introduction of an enhancer element. PMID- 2983710 TI - Nuclear testicular 1,25-dihydroxyvitamin D3 receptors in Sertoli cells and seminiferous tubules of adult rodents. AB - 1,25(OH)2D3 receptors were studied in whole testes, Sertoli cells, seminiferous tubules, Leydig cells and spermatogonia of adult NMRI mice and SD rats. Specific reversible high affinity binding (KD 1.4 x 10(-10)M; Nmax 72 fmol/mg protein) by a 3.5 S macromolecule was demonstrated in whole testes, Sertoli cells and seminiferous tubules. With identical techniques, no receptors were found in Leydig cells despite previous reports of 1,25(OH)2D3 actions on Leydig cell function. PMID- 2983709 TI - Substrate specificities of insulin and epidermal growth factor receptor kinases. AB - The abilities of insulin and EGF stimulated protein kinases to phosphorylate a series of exogenous substrates were compared using wheat germ lectin purified preparations of solubilized rat liver membranes. Three different kinds of substrates were found: substrates phosphorylated primarily by insulin stimulated kinase, substrates phosphorylated primarily by EGF stimulated kinase and substrates phosphorylated by both kinases to a similar extent. These results indicate that the insulin and the EGF receptor kinase have different, but overlapping, substrate specificities. In vivo, phosphorylation of cellular proteins by various hormone receptor kinases may be part of the signal transmission process for actions of the hormones. Different substrate specificities of kinases of different hormone receptors may therefore represent an important mechanism to preserve the specificity of the hormonal signal at the post receptor level. PMID- 2983711 TI - High-affinity calcium-binding proteins in Escherichia coli. AB - Crude extracts of Escherichia coli contain at least three heat stable proteins of Mr, 33,000, 47,000, and 60,000, which bind 45Ca2+ in buffers containing micromolar calcium and physiological salt concentrations. Fractions containing these proteins neither activated the calmodulin-dependent enzyme, NAD kinase, nor inhibited the activity of this enzyme in the presence of brain calmodulin. Radioimmunoassay of crude extracts for calmodulin indicated the presence of a calmodulin-like antigen. Crude extracts also contain proteins that interact with 2-trifluoromethyl-10H-(3'-aminopropyl)phenothiazine-Sepharose in a calcium dependent manner, but proteins eluted from this resin did not bind calcium with high affinity. PMID- 2983712 TI - Tumor promoter phorbol 12-myristate, 13-acetate inhibits phosphoinositide hydrolysis and cytosolic Ca2+ rise induced by the activation of muscarinic receptors in PC12 cells. AB - Preincubation of PC12 cells (used both before and after differentiation by NGF) with phorbol myristate acetate (PMA) was without effect on the basal concentration of inositol phosphates (metabolites of phosphoinositide hydrolysis) and of free cytosolic Ca2+, but inhibited considerably the increases induced by the cholinergic agonist carbachol via the activation of the muscarinic receptor. Inasmuch as binding was unaffected, this inhibition might occur at the level of receptor coupling to its transduction mechanism(s). Inhibition appeared within 1 min and was maximal after 3 min. The concentrations of PMA needed (10(-9)-10( 8)M) were in the range believed to cause specifically the activation of protein kinase C. The muscarinic receptor, via the hydrolysis of phosphoinositides and the generation of diacylglycerol, participates in the regulation of the latter enzyme. Our data suggest therefore that the receptor operates under stringent feedback control by the metabolites generated as a consequence of its activation. PMID- 2983713 TI - Modulation of adenylate cyclase by guanine nucleotides and Kirsten sarcoma virus mediated transformation. AB - Certain tumour cells contain activated ras genes that code for 21 000 dalton proteins (p21). These proteins associate with the inner face of the plasma membrane and bind guanine nucleotides specifically. In order to determine whether p21s have functions similar to other GTP binding proteins, we investigated the regulation, by guanine nucleotides, of adenylate cyclase (AC) activity in membrane preparations isolated from fibroblasts (C127) transformed by a temperature sensitive mutant of Kirsten sarcoma virus (Ts 371). The degree of AC stimulation by GMP P(NH)P increased when these cells were shifted from the permissive temperature (33 degrees C) to the non-permissive temperature (39 degrees C). This effect was more pronounced at low Mg++ and low GMP P(NH)P concentrations. AC stimulation remained unchanged in rat fibroblasts infected with a temperature sensitive mutant of Rous Sarcoma virus. AC activity was depressed in C127 cells infected with wild type KiMSV. Our data illustrate the feasibility of correlating alterations in the AC system with ras gene expression and using such experimental approaches to elucidate the physiological functions of the p21 proteins. PMID- 2983714 TI - Cyclic AMP inhibits platelet activation independently of its effect on cytosolic free calcium. AB - Stimulation of platelets with the ionophore A23187, thrombin, ADP or PAF-acether resulted in a rapid increase of cytosolic free Ca2+, as measured with Quin-2, and in aggregation, 5HT secretion and - in the case of the first two agonists - thromboxane generation. PGI2 and dibutyryl cyclic AMP inhibited all these responses, except in the case of A23187, in response to which the increase in Ca2+ was unaffected, although the other responses were inhibited. The inhibition of aggregation and secretion in response to the combination of thrombin and A23187 was indistinguishable from that in response to thrombin alone. It thus appears that cAMP inhibits these responses independently of its effect in lowering cytosolic free Ca2+. PMID- 2983715 TI - Measurement of free radical oxygen generation by cytochrome c reduction requirement for cytochrome c oxidase blockade. AB - Data is presented showing that one commercial preparation of cytochrome c, used to trap and measure free radical superoxide anion, can be contaminated with cytochrome c oxidase activity. This activity can vary from lot to lot, can introduce variability into the measurement of superoxide anion and can result in falsely low estimations of free radical formation. This cytochrome c oxidase activity can be inhibited by low (0.2 mM) concentrations of KCN. Blockade of the cytochrome c oxidase activity allows reproducible measurement of superoxide anion formation at low levels by red cells. PMID- 2983716 TI - The association of calmodulin with subcellular fractions isolated from rat liver. AB - Calmodulin associated with rat liver mitochondria has been found to belong to a contaminant membranous fraction which contains different subcellular membranes. The concentration of calmodulin in this fraction is relatively high, about 1.6 micrograms/mg protein, and can not be decreased with EGTA. The calmodulin-rich membranous fraction seems to contain cytoskeletal proteins which could be responsible for the binding of calmodulin. PMID- 2983717 TI - The reactive cysteine residue of pig kidney fructose 1,6-bisphosphatase is related to a fructose 2,6-bisphosphate allosteric site. AB - Modification of a highly reactive cysteine residue of pig kidney fructose 1,6 bisphosphatase with N-ethylmaleimide results in the loss of activation of the enzyme by monovalent cations. Low concentrations of fructose 2,6-bisphosphate or high (inhibitory) levels of fructose 1,6-bisphosphate protect the enzyme against the loss of monovalent cation activation, while non-inhibitory concentrations of the substrate gave partial protection. The allosteric inhibitor AMP markedly increases the reactivity of the cysteine residue. The results indicate that fructose 2,6-bisphosphate can protect the enzyme against the loss of potassium activation by binding to an allosteric site. High levels of fructose 1,6 bisphosphate probably inhibit the enzyme by binding to this allosteric site. PMID- 2983718 TI - Chlordiazepoxide is a competitive thyrotropin-releasing hormone receptor antagonist in GH3 pituitary tumour cells. AB - Addition of thyrotropin-releasing hormone (TRH) to [3H]-inositol pre-labelled GH3 pituitary tumour cells suspended in medium containing 10mM lithium chloride led to a rapid diminution in cellular [3H]-inositol and increase in [3H]-inositol 1 phosphate (InslP), [3H]-inositol bisphosphate (InsP2) and [3H]-inositol trisphosphate (InsP3). In the presence of the benzodiazepine tranquillizer, chlordiazepoxide, the TRH concentration-response curves for these effects were shifted to the right in a parallel fashion. The Ki for chlordiazepoxide in inhibiting all four responses was 1.5 X 10(-5)M. Chlordiazepoxide did not inhibit the small bombesin-induced rise in [3H]-InslP. Another benzodiazepine, diazepam, was less active. The TRH-induced rise in cytosolic free calcium monitored in Quin 2-loaded GH3 cells was also blocked by chlordiazepoxide in a competitive manner, while that induced by high K+-induced depolarisation was unaffected. It is suggested that chlordiazepoxide acts as a competitive antagonist at the level of the TRH receptor. PMID- 2983719 TI - Unexpected specificity in the thioredoxin activation of fructose-bis-phosphatases from different plants. AB - Green seedlings of soy bean and wheat contain, like the plant seeds, multiple thioredoxin proteins which possess all typical thioredoxin properties but are inactive in the stimulation assay with spinach fructose-bis-phosphatase. However the pure proteins do have thioredoxin f activity when tested with homologous enzymes isolated from soy bean or wheat leaves, respectively, in the presence of Mg++. This new type of species specificity, unknown in all other in vitro assays of reduced thioredoxins, has to be considered in characterizing complete thioredoxin profiles in plants. PMID- 2983720 TI - Distinct atrial natriuretic factor receptor sites on cultured bovine aortic smooth muscle and endothelial cells. AB - Cultured bovine aortic smooth muscle and endothelial cells each display distinct specific binding sites for radiolabeled atrial natriuretic peptide (ANF). 125I pro-rANF (103-126)I binding to both cell types is rapid, reversible and competitive. Scatchard plots of the binding data show Bmax values of 5.5 and 0.1 2.1 X 10(5) sites/cell and Kd values of 2.1 and 0.3 nM for smooth muscle and endothelial cells, respectively. In addition, ANF elevates levels of cGMP substantially in both cell types at concentrations of ANF close to its Kd and Ki for binding. Sodium nitroprusside, however, has essentially no effect on cGMP levels in either cell type. These results show that distinct functionally active receptor sites for ANF exist on both vascular smooth muscle and endothelial cells. PMID- 2983721 TI - Potential role for a guanine nucleotide regulatory protein in chemoattractant receptor mediated polyphosphoinositide metabolism, Ca++ mobilization and cellular responses by leukocytes. AB - Islet activating protein from Bordetella pertussis toxin which ribosylates certain guanine nucleotide regulatory proteins causes a marked reduction of chemoattractant-elicited responses such as chemotaxis, O2 production and cAMP elevations in human polymorphonuclear leukocytes. The toxin appears to exert its effects by preventing the rapid breakdown of phosphatidylinositol 4,5 bisphosphate induced by the chemotactic peptide N-formyl-methionyl-leucyl phenylalanine, thereby inhibiting the increase in intracellular [Ca++] which normally follows chemoattractant stimulation. Responses of leukocytes exposed to Concanavalin A, the Ca++ ionophore A23187, or phorbol myristate acetate were not affected by the toxin. Thus the chemoattractant receptor appears to be coupled to a phosphoinositide specific phospholipase C through a guanine nucleotide regulatory protein. We propose that this complex of receptor-guanine nucleotide regulatory protein-phospholipase C may be applicable to the class of receptors which mobilize intracellular Ca++ by stimulating polyphosphoinositide breakdown. PMID- 2983722 TI - Phosphorylation of fodrin (nonerythroid spectrin) by the purified insulin receptor kinase. AB - Fodrin (nonerythroid spectrin) from porcine brain was found to be phosphorylated on tyrosine residues by the purified insulin receptor kinase. The phosphorylation occurred in an insulin-sensitive manner with a physiologically relevant km. The beta(235 K) subunit of fodrin, but not the alpha(240 K) subunit, was phosphorylated by the kinase. Neither the alpha(240 K) subunit nor the beta(220 K) subunit of erythrocyte spectrin was phosphorylated under the same conditions. Fodrin phosphorylation by the purified insulin receptor kinase was markedly inhibited by F-actin. These data raise the possibility that tyrosine phosphorylation of fodrin plays some roles in the regulation of plasma membrane microfilament interaction. PMID- 2983723 TI - Sialic acid moiety is responsible for the charge heterogeneity and the biological potency of rat lutropin. AB - To elucidate a possible role of sialic acid moiety in the electrical heterogeneity of rat pituitary lutropin, seven components separated were individually treated with neuraminidase. Some intermediates with isoelectric points corresponding to the native components were concomitantly seen at the serial stages of the enzyme treatment. All the treated components showed an isoelectric point of about 10.0 which was the same to the isoelectric point of one of the seven components. Desialylation of the components with less biological activity caused enhancement of the in vitro cyclic AMP producing- and testosterone producing-activities as well as the binding activity to the receptor. It is concluded that the number of sialic acid moiety in lutropin is responsible for the charge heterogeneity and the biological potency of the hormone. PMID- 2983724 TI - Partial purification and functional reconstitution of GDP sensitive brown adipose tissue mitochondria uncoupling protein using octyl glucoside. AB - A partial purification of the uncoupling protein of brown adipose tissue mitochondria (BATM) was achieved by extraction of BATM with 40 mM octyl glucoside, followed by affinity chromatography on ATP-agarose. The isolated protein was functionally reconstituted into liposomes using octyl glucoside dialysis. Proteoliposomes containing the uncoupling protein had an increased proton or chloride conductance when subjected to a valinomycin-induced potassium diffusion potential. The increased ion conductance was consistently found to be inhibited by 200 microM GDP. PMID- 2983725 TI - Detection of caldesmon in muscle and non-muscle tissues of the chicken using polyclonal antibodies. AB - Polyclonal antibodies raised in rabbits against chicken gizzard caldesmon have been purified and used in immunoblotting experiments to study the distribution of this actin- and calmodulin-binding protein in diverse tissues of the chicken. Total homogenates and heat-treated homogenate supernatants derived from each tissue were subjected to sodium dodecyl sulfate-polyacrylamide gradient slab gel electrophoresis and immunoblotting using the horseradish peroxidase method. All chicken tissues examined contained caldesmon of Mr = 141,000. The amount of caldesmon found in the different tissues varied considerably and semi quantitative comparison of stained immunoblots indicated the following relative caldesmon contents: gizzard greater than oesophagus greater than duodenum = small intestine greater than lung greater than aorta greater than heart = skeletal muscle greater than kidney = trachea greater than brain greater than liver. Each tissue revealed small amounts of lower Mr immunoreactive proteins, predominantly bands of Mr 94,000 and 70,000, which appear to be proteolytic fragments of caldesmon. Isolated caldesmon was found to be highly sensitive to proteolysis. The widespread distribution and similarity of caldesmon in different tissues of the chicken suggest its functional importance and structural conservation. PMID- 2983726 TI - A novel cyclic opioid peptide analog showing high preference for mu-receptors. AB - The side-chain to side-chain cyclized opioid peptide analogs H-Tyr-D-Orn-Phe-Asp NH2 (I) and H-Tyr-D-Lys-Phe-Glu-NH2 (II) were synthesized and tested in the guinea pig ileum and mouse vas deferens assays and in binding assays based on displacement of mu- and delta-opioid receptor-selective radioligands from rat brain membranes. The more rigid cyclic analog I containing a 13-membered ring structure showed very high preference for mu-receptors over delta-receptors, whereas the more flexible cyclic peptide II (15-membered ring) was non-selective. These results indicate that variation in the degree of conformational restriction of opioid peptides can produce drastic shifts in their receptor selectivity profile. Because of its high mu-receptor selectivity and rigidity cyclic analog I will be useful for determining the conformational requirements of mu-opioid receptors. PMID- 2983727 TI - Dibromothymoquinone (DBMIB)-mediated reduction of cytochrome in solution and formation of DBMIB-cytochrome complex. AB - Addition of dibromothymoquinone (DBMIB) to cytochrome f and cytochrome c in solution caused a reduction of the cytochromes. DBMIB formed a complex with cytochrome at a stoichiometry of 2 to 3 DBMIB per cytochrome molecule. These observations suggest a possibility that DBMIB acts at or near the function of cytochrome f in the photosynthetic membranes. PMID- 2983728 TI - Human myeloid leukemia cells metabolize 25-hydroxyvitamin D3 in vitro. AB - Human promyelocytic leukemia cells (HL-60) converted 25-hydroxyvitamin D3 to two more polar metabolites during in vitro incubations. A two-step high pressure liquid chromatography system revealed two unique elution positions of those leukemic cell-derived metabolites that exactly co-migrated with the elution positions of 5(Z)-19-nor-10-oxo-25-hydroxyvitamin D3 and 5(E)-19-nor-10-oxo-25 hydroxyvitamin D3, respectively. These unique metabolites did not bind specifically to a protein receptor for 1,25-dihydroxyvitamin D3. PMID- 2983729 TI - Inhibition of analgesia by C-terminal deletion analogs of human beta-endorphin. AB - Human beta-endorphin (beta h-EP) analogs of variable chain lengths have been investigated for their potency in inhibiting analgesia induced by beta h-EP or by the potent opiate etorphine. It was found that beta h-EP-(1-28) inhibits the analgesic effect of beta h-EP and etorphine when co-injected intracerebroventricularly into mice. Antagonism by competition at same opioid receptor subtypes is suggested from parallel shifts of the dose-response curve of etorphine or beta h-EP in the presence of increasing doses of beta h-EP-(1-28). On a molar basis, beta h-EP-(1-28) is nearly 10 times more potent than naloxone. The reduction of the chain length from residues 1-28 to 1-27 lowered the antagonist potency while further reduction of the peptide chain led to a complete loss of inhibitory activity. From comparison of the opioid-receptor binding affinity, analgesic activity and antagonist potency, it is concluded that the C terminus of beta-EP is critical to the biological efficacy of the molecule and that the antagonist activity of C-terminal deletion analogs is probably mediated through residues 27 and 28. PMID- 2983730 TI - Aggregation state of the gonadotropin receptor. AB - The aggregation state of the gonadotropin receptor has been examined by coupling fluorescence energy transfer donor and acceptor fluorophores to hCG and LH. Energy transfer is observed at low (4 degrees C) but not at high (37 degrees C) temperature. Energy transfer could also be detected with receptor solubilized in the presence of hormone at the lower temperature only. Solubilization of receptor in the absence of hormone and subsequent addition of hormone conjugates revealed no energy transfer. These results are consistent with stabilization of receptor complexes at low temperatures, but presumptive hormone induced receptor dissociation under physiological conditions. PMID- 2983731 TI - Evidence for a decrease in the efficiency of beta-receptor coupling to adenylate cyclase in liver membranes from sucrose-fed rats. AB - Sucrose feeding has been shown previously to alter the plasma concentration of several factors which may regulate beta-adrenergic receptors, including corticosteroids and insulin as well as altered sympathetic nervous system (SNS) tone. For this reason we initiated a study of the effects of sucrose feeding on the beta-adrenergic receptor-adenylate cyclase system in rat liver plasma membranes. Beta-Adrenergic responsiveness was monitored by measuring isoproterenol stimulation of adenylate cyclase activity, while beta-adrenergic receptor characteristics were evaluated by analyzing [125I]iodocyanopindolol [( 125I]CYP) binding. Rats fed rat chow ad lib. supplemented by drinking water containing 10% sucrose solution exhibited a 50-75% reduction in hepatic isoproterenol-sensitive adenylate cyclase activity. This effect of sucrose was also observed in adrenalectomized (ADX) and 6-hydroxydopamine-pretreated animals, ruling out a causal role for corticosteroids or the sympathetic nervous system respectively. No effect was observed on basal, glucagon-, fluoride- or GTP stimulated adenylate cyclase. A small but significant decrease in [125I]CYP specific binding capacity was observed in liver membranes prepared from sucrose fed ADX rats, whereas no change in [125I]CYP binding capacity was observed in in sucrose-fed normal rats. These observations suggest that beta-receptor to adenylate cyclase coupling efficiency is decreased by the sucrose diet. The activities of two membrane-associated phospholipid methyltransferases and the content of endogenous S-adenosylmethionine in liver were reduced by sucrose feeding, implying a defect in the methylation pathway for phosphatidylcholine synthesis. The possible relationship between this latter finding and the observed decrease in beta-adrenergic receptor to adenylate cyclase coupling efficiency is discussed. PMID- 2983732 TI - Dual effect of isoprenaline on glucose transport and response to insulin in isolated adipocytes. AB - Glucose transport as assessed by the uptake rate of 3-O-methylglucose was stimulated in isolated rat fat cells by preincubation with isoprenaline or orciprenaline. The effect was apparently mediated by beta 1-receptors, since (1) it was abolished by propranolol, (2) it closely paralleled the stimulation of lipolysis, and (3) isoprenaline was 10(2) times more potent that orciprenaline. Isoprenaline enhanced the effect of submaximal insulin concentrations as well as the basal transport rate but failed to increase the maximal effect of insulin. The stimulatory effect of isoprenaline was antagonized by adenosine deaminase which removes adenosine spontaneously released from the cells, and by bordetella toxin (IAP) which blocks the inhibitory coupling component of adenylate cyclase. Moreover, bordetella toxin uncovered an inhibitory effect of isoprenaline on insulin stimulated glucose transport. There was no apparent correlation between the effects on glucose transport and the response of cellular cyclic AMP levels to the agents investigated. It is suggested that a step in the coupling of beta receptors and adenylate cyclase, but not total cellular cyclic AMP levels, may mediate stimulatory as well as inhibitory effects of catecholamines on glucose transport in the adipocyte. PMID- 2983733 TI - Interaction of local anaesthetics with cytochrome oxidase studied with fluorescence quenching. AB - The interaction of a series of eight local anaesthetics with cytochrome oxidase chosen as a membrane model protein has been studied with fluorescence technique using quinacrine as a fluorescent probe. The existence of hydrophobic interactions with a non polar region of cytochrome oxidase complex has been shown. The ability of the drug molecules to displace quinacrine bound to cytochrome oxidase correlate as closely with their anaesthetic potency as with their octanol-water partition coefficient. Our results are in good agreement with a recent model of local anaesthetic action on nerve membranes presenting a site of anaesthesia including both lipid binding and protein binding environments. PMID- 2983734 TI - Free radical production from normal and adriamycin-treated rat cardiac sarcosomes. AB - The production of hydroxyl radicals in rat myocardial sarcosomes treated with adriamycin was demonstrated by the electron spin resonance technique of spin trapping. Using the spin trapping agent 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), the formation of a hydroxyl radical spin adduct was observed in adriamycin treated rat heart sarcosomes with NADPH as co-factor. Oxygen, NADPH and sarcosomal protein were absolute requirements for hydroxyl radical production. Hydroxyl radical spin adduct formation was not inhibited by the metal ion chelators diethylenetriaminepenta-acetic acid (DETAPAC) or desferrioxamine, or by addition of superoxide dismutase but could be inhibited by addition of catalase and high concentration of the hydroxyl radical scavengers mannitol and N acetylcysteine. Hydroxyl radical production in adriamycin-treated rat myocardial sarcosomes appears to arise from the reductive metabolism of adriamycin by an NADPH-dependent quinone reductase--NADPH: cytochrome P450 reductase; the reduced quinone (semiquinone) reduces oxygen to hydrogen peroxide, probably via superoxide, although this was not detected. The hydrogen peroxide appears to react directly with adriamycin semiquinone, although involvement of traces of iron in a Fenton type of reaction cannot be excluded. From the observations it is suggested that adriamycin-induced cardiotoxicity is an oxidative pathology arising from intracellular generation of relatively high levels of hydroxyl radicals. PMID- 2983736 TI - Can ribonucleotide reductase be considered as an effective target for developing antiherpes simplex virus type II (HSV-2) compounds? AB - Herpes simplex viruses are known to induce virus specified ribonucleotide reductase (RR) in infected cells. RR is considered as a possible target for the development of antiviral agents. In this study, the role of RR in virus replication has been investigated. The sensitivity of RR to hydroxyurea (HU) from virus infected and uninfected HeLa S3 cells was similar with IC50 values of 0.12 and 0.14 mM. In the presence of 2 mM HU, and 10 microM tetrahydrouridine (THU), a cytidine deaminase inhibitor, the incorporation of [14C]cytidine into viral DNA was found to be inhibited by 95%; [32P]-incorporation into viral DNA under the same conditions was inhibited by 75%. The pool size of dCTP and dGTP was 50 and 70%, respectively, with no significant effect on dATP and dTTP pools in virus infected cells treated with 2 mM HU, as compared with virus infected cells receiving no drug treatment. HU at 2 mM could not inhibit HSV-2 yield by more than one log. These results suggest that virus RR is not an effective target for developing anti HSV-2 compounds. PMID- 2983735 TI - Effect of chronic desmethylimipramine or electroconvulsive shock on selected brain and platelet neurotransmitter recognition sites. AB - Rats were treated with electroconvulsive shock (ECS), desmethylimipramine (DMI), ECS plus DMI, or diazepam. In vitro analyses showed that chronic ECS produced an elevated density of recognition sites for [3H]imipramine (IMI) in platelet membranes, but had no effect on membrane preparations derived from cortical tissue. A similar elevation in receptor binding was seen exclusively in platelets after chronic ECS plus DMI, whereas no effect was observed with DMI alone. Equilibrium dissociation constant (KD) values for [3H]IMI were also increased in platelet membranes from rats given chronic ECS or ECS plus DMI treatment. Chronic ECS or DMI administration produced a decreased density of beta-adrenergic recognition sites in frontal cortex and cerebellum as assessed by [3H]dihydroalprenolol (DHA) binding. The combination of ECS plus DMI produced a similar decrease. In addition, chronic diazepam administration produced a down regulation of the beta-adrenergic receptor only in the cerebellum. These data provide evidence for the differential regulation of brain and peripheral neurotransmitter recognition sites. PMID- 2983737 TI - Preparation and characterization of subcellular fractions from the head kidney of the Northern pike (Esox lucius), with particular emphasis on xenobiotic metabolizing enzymes. AB - The present study was designed to prepare and characterize subcellular fractions from the head kidney of the Northern pike (Esox lucius), with special emphasis on the preparation of a microsomal fraction suitable for studying xenobiotic metabolism. The purity of the different fractions obtained by differential centrifugation as well as the recovery of different cell components was determined using both enzyme markers and morphological criteria. Finally, the subcellular distributions of several drug-metabolizing enzymes (NADPH-cytochrome c reductase, NADH-ferricyanide reductase, glutathione transferase, epoxide hydrolase) were determined. With the exception of NADPH-cytochrome c reductase, the subcellular distributions obtained here for drug-metabolizing and marker enzymes closely resembled those reported for rat liver. NADPH-cytochrome c reductase was apparently partially solubilized here from microsomal vesicles by an endogenous protease, which reduced its usefulness as a marker enzyme and raises questions concerning the measurement of activities catalyzed by the cytochrome P-450 system in these subfractions. In other respects the microsomal fraction prepared here from the pike head kidney seems well-suited for studies of drug metabolism. PMID- 2983738 TI - Rate-limiting steps in isoproterenol and forskolin stimulated lipolysis. AB - Using the flask-incubated fat cell system, effects of isoproterenol and forskolin on glycerol release, cyclic AMP levels and protein kinase were studied. Isoproterenol increased cyclic AMP levels, protein kinase activity and glycerol release over the same concentration range (10(-9) M to 10(-6) M). Forskolin also increased all three variables in a concentration-dependent manner (10(-7) M to 10(-4) M). The maximum response for each variable was significantly greater with forskolin than with isoproterenol. A combination of isoproterenol and forskolin resulted in an additional increase in cyclic AMP over forskolin alone, but no significant increase in protein kinase activity or glycerol release. These results support the concepts that the maximum lipolytic response to isoproterenol is limited by the accumulation of cyclic AMP and the maximum lipolytic response to forskolin is limited by some step distal to cyclic AMP production, possibly activation of protein kinase. At high concentrations of forskolin or with a combination of forskolin and isoproterenol, cyclic AMP levels were in excess of those needed to maximally activate protein kinase and lipolysis. PMID- 2983739 TI - Autoantibody specificities of immune complexes sequestered in articular cartilage of patients with rheumatoid arthritis and osteoarthritis. AB - To define autoantibody specificities of immune complexes sequestered in articular cartilage of patients with rheumatoid arthritis and osteoarthritis, extracts were obtained from articular cartilage specimens from 16 patients with rheumatoid arthritis, 11 patients with osteoarthritis, and 6 normal controls. Radioimmunoassays of the extracts revealed that rheumatoid cartilage contained 37 times more IgM and 14 times more IgG than did normal cartilage extracts. In addition, osteoarthritic cartilage contained 3 times more IgM and IgG than the normal tissues. IgM rheumatoid factor was found in 13 of 16 rheumatoid cartilage extracts but in none of 11 osteoarthritic or 6 normal control extracts. IgG rheumatoid factor was detected in 4 of 7 seropositive rheumatoid but in none of 5 osteoarthritic cartilage extracts. More than 60% of the rheumatoid cartilage extracts were positive for native and denatured collagen II antibodies. Surprisingly, 50% of the osteoarthritic specimens also contained significant titers of collagen antibodies. Similar results were obtained with osteoarthritic menisci extracts. These findings indicate that the immune complexes sequestered in rheumatoid cartilage contain autoantibodies that are probably synthesized locally by cells infiltrating the inflamed synovium. If immune complexes trapped in cartilage play an important role in cartilage damage, our findings would provide a possible pathogenic mechanism that explains the self-perpetuating and chronic nature of cartilage degradation in rheumatoid arthritis and osteoarthritis. PMID- 2983740 TI - A subset of rheumatoid arthritis patients with a pattern of Epstein-Barr virus antibodies similar to that found in primary and secondary immunodeficiency diseases. PMID- 2983741 TI - N-ethylmaleimide as oxidizing agent in biological and non-biological systems. AB - The hypothesis that N-ethylmaleimide (NEM) may function as an oxidizing agent in biological and non-biological systems, has been tested. Spectrophotometric determination of cytochrome a-redox-state have revealed that NEM promotes the transition of this respiratory chain component in a more oxidated state. To overcome the possibility that the NEM effect may be determined by the inhibition on primary dehydrogenase, duroquinol (QH2) has been used as substrate in the presence of rotenone and malonate. The stimulation of cytochrome a-oxidation is correlated to the one on the QH2 oxidation determined by following the formation of duroquinone. In the absence of any biological system, spontaneous oxidation of ferrocyanide and TMPD is greatly increased by NEM. The differential stimulation induced by maleimide and succinimide indicate that the oxidizing effect of NEM may be considered a chemico-physical property of its molecule mainly due to the presence of a double bond. It is proposed that besides a sulfhydryl reagent, NEM behaves as an oxidizing agent with an interacting site in the region QH2 cytochrome a of the respiratory chain. PMID- 2983742 TI - An indirect and highly sensitive method for the determination of the flow of reducing equivalents in the respiratory chain. AB - Reactions leading to oxido-reduction of TMPD have shown that, in its oxidized form, this compound has among others an extinction maximum at 610 nm; with the exception of cytochrome a, at this wavelength none of the respiratory chain intermediates has the ability to absorb the incident light. This property together with the one of reacting with cytochrome c, has given us the possibility to use TMPD as a "probe" of the reducing equivalents flow in the respiratory chain. Added to mitochondrial suspension, TMPD undergoes redox cycles in relation to the activity of the respiratory chain, modulated by increasing concentrations of succinate or respiratory inhibitors. NEM-induced reversible oxidation on the respiratory intermediates can also be determined by following the TMPD oxidation. The preliminary data obtained are thus consistent with the hypothesis that in appropriate conditions, the TMPD redox state can be used as a probe of the respiratory chain activity. PMID- 2983744 TI - [Prolactin after baclofen in healthy subjects and prolactinoma patients]. AB - Serum prolactin (PRL) levels in basal conditions (two samples) and 30, 60, 90, 120, 150 e 180 minutes after oral administration of baclofen (20 mg) were evaluated in 6 healthy subjects and in 10 patients with prolactinoma. The effect of baclofen (20 mg by mouth) on the PRL secretion cimetidine (400 mg i.v.) or domperidone (20 mg i.v.) induced were evaluated in 9 healthy women by administration of baclofen 60 minutes before cimetidine or domperidone. Baclofen was unable to significantly rise serum PRL levels in healthy subjects and in patients affected by prolactinoma and furthermore did not interfere with PRL rise domperidone induced. On the contrary baclofen decreased PRL rise cimetidine induced. It was concluded that: in basal condition, GABAb receptor don't play an obvious role in modulation of PRL secretion; when H2 istaminergic inhibition on PRL secretion is blocked (at an hypothalamic site), a GABA inhibition, b receptor mediated, on PRL secretion became more clear; the domperidone blockade of hypophysial dopaminergic receptors suggests that GABAb modulation of prolactin secretion don't obtain itself by dopaminergic pathways. PMID- 2983743 TI - [Growth hormone after baclofen in normal and acromegalic subjects]. AB - Serum growth hormone (GH) levels in basal conditions (two samples) and 30, 60, 90, 120, 150 and 180 minutes after oral administration of baclofen (20 mg) were evaluated in 6 healthy subjects and in 6 acromegalic patients. The effect of cimetidine (400 mg i.v.) administrated 45 minutes after baclofen (20 mg by mouth) were evaluated in 9 healthy women. Baclofen was able to significantly rise serum GH levels in normal subjects but no in acromegalic patients. Cimetidine suppress GH increase induced by baclofen. It was concluded that: 1) baclofen, GABAb receptor agonist, stimulate GH secretion by inhibition of GIF secretion or by stimulation of GRF secretion; 2) istamine, through H2 receptors in the hypothalamus, is important to mediate GH release induced by stimulation of GABAb receptors. PMID- 2983745 TI - [Effect of the globus pallidus on hippocampal epilepsy in the cat]. AB - The action of the pallidum on electrically induced afterdischarge of the hippocampus (HAD) was studied. Significant facilitatory influences on HAD duration were observed when pallidal conditional stimulation immediately preceded hippocampal test stimulation. The phenomenon appeared to be into correlation with the interval between conditioning and test stimulation. Experimental data are discussed as strongly presumptive of a functional interrelationship between the inhibitory role played by the caudate on both pallidum and hippocampus; facilitatory influence of the pallidum on HAD duration is discussed too. PMID- 2983746 TI - [Weak chemiluminescence from infarcted rat brain and basic studies on the generation of chemiluminescence]. AB - Chemiluminescence measurement in vivo system and its related studies on free radical reaction were reported. Weak chemiluminescence from rat brain surface subjected to ischemic cerebral stroke could be detected. It was observed just after embolization and lasted at least four hours. In order to investigate the mechanism of the generation of chemiluminescence and a side reaction of lipid peroxidation, several studies were made using 10% brain homogenate under oxygenated conditions at 37 degrees C. Brain homogenate showed increasing chemiluminescence that reached a plateau level a few hours later. Chemiluminescence, accompanying with thiobarbituric acid reactive substance (TBARS) formation, was based on free radical reaction with requirement of oxygen. Time course increase in TBARS formation, however, revealed some differences from that of integrated light intensity. Chemiluminescence spectrum in the visible region showed that excited indole chemicals (triplet states) generated in the present system emitted light during their return to ground state, but not singlet molecular oxygen. Suspension of acetone powder in Tris-HC1 buffer prepared from brain homogenate also disclosed chemiluminescence to some extent, whereas liposomes made of extracted brain lipid did not. These results probably indicate that proteins are essential for luminescence, but not lipid only, and involves the lipid-protein interaction in oxygenated brain homogenate. In the process of lipid-protein interaction, neither fatty acids composition nor protein molecules significantly altered, except for a newly-appeared high molecular weight protein. On the other hand, SH group was suggested to be quite vulnerable to free radical attack.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983747 TI - Carbon dioxide laser surgery for benign oral lesions. PMID- 2983748 TI - The effects of dazoxiben, an inhibitor of thromboxane synthetase, on cold-induced forearm vasoconstriction and platelet behaviour in different individuals. AB - The effect of dazoxiben, a thromboxane synthetase inhibitor, on cold-induced forearm vasoconstriction was determined in two groups of human volunteers, those in whom dazoxiben abolished the platelet aggregation and release reaction induced by sodium arachidonate (group I) and those in whom it did not (group II). Dazoxiben abolished cold-induced forearm vasoconstriction in group I volunteers but not in those of group II. These results imply a correlation between platelet behaviour and cold-induced changes in vascular tone. In the group I volunteers the effect of dazoxiben on cold-induced vasoconstriction was abolished by 1800 mg of aspirin, but not by 40 mg. Since the lower dose of aspirin inhibits platelet cyclo-oxygenase but has no effect on cyclo-oxygenase in blood vessel walls, it is possible that platelets play no part in the modulation of vascular tone by dazoxiben. It is more likely that the effects of dazoxiben are confined to the vessel wall. PMID- 2983749 TI - Cardiac effects of sodium stibogluconate. AB - Sodium stibogluconate although potentially cardiotoxic is the drug of choice for Kalaazar and cutaneous leishmaniasis due to Leishmania braziliensis. Increasing use of this drug in the British Army has necessitated a formal evaluation of its cardiac side-effects. Consequently a detailed study of the cardiac effects of sodium stibogluconate was undertaken in 22 male soldiers using for the first time modern non-invasive techniques. Intravenous sodium stibogluconate 600 mg daily for 10 days did not affect blood pressure, heart rate, left ventricular contractile function or rhythm. Electrocardiography showed a reversible reduction of T wave amplitude. PMID- 2983750 TI - Lupus pernio: a clinico-radiological study of thirty-five cases. AB - Thirty-five patients with lupus pernio were observed in a series of 818 patients with clinical and histological evidence of sarcoidosis. This analysis provides the natural history of lupus pernio and its associated clinico-radiographic features. There was intrathoracic involvement in 74% of patients, upper respiratory tract disease in 54%, reticulo-endothelial involvement in 54%, bone cysts in 43% and ocular lesions in 37%. It predominated in West Indies-born women. Histological confirmation was easily obtained by nasal mucosal biopsy and this could be repeated serially to evaluate the response to various treatments. Nasal bone radiographs showed destruction of the nasal bones, porosis and alteration in bone texture. Peripheral bone cysts were evident in 50% of patients who were radiographed, and in about 50% of these patients, the cysts were found in both hands and feet. PMID- 2983751 TI - Mortality of workers in a French asbestos cement factory 1940-82. AB - The mortality of a complete cohort of 1506 French asbestos cement workers employed for at least five years is related to the time elapsed since first exposure. The mortality from all causes (analysed by the "man-years method") has been found to be above normal only in those subjects employed for more than 20 years, with more than 35 years of follow up. Standardised mortality ratios for cancers of all sites (ICD 140-209) and pulmonary cancer (ICD 162-163.0) have been assessed in subjects whose first exposure dates go back more than 20 years. Mortalities from cancer of all sites and from pulmonary cancer have been detected in excess in workers employed for more than 20 years and originally hired when aged 25 or under. PMID- 2983752 TI - A spin label study of the effects of asbestos, quartz, and titanium dioxide dusts on the bovine erythrocyte membrane. AB - The effects of five UICC asbestos samples, titanium dioxide, and quartz on the bovine red cell membrane have been studied in erythrocyte ghosts by the spin labelling technique. Analysis of the electron paramagnetic resonance (EPR) spectra of two sulphydryl reactive spin labels and one fatty acid spin label in red cell ghosts showed modifications in membrane protein after asbestos treatment but no alterations in membrane lipids. In experiments with quartz no membrane changes were noted but titanium dioxide altered the proteins bound with the protein reactive spin label used in the present study. The possible mechanism for these effects is discussed. PMID- 2983753 TI - Iron(III)-adriamycin and Iron(III)-daunorubicin complexes: physicochemical characteristics, interaction with DNA, and antitumor activity. AB - Fe(III) complexes of two anthracyclines, adriamycin and daunorubicin, have been studied. Using potentiometric and spectroscopic measurements, we have shown that adriamycin and daunorubicin form two well-defined species with Fe(III), which can be formulated as respectively Fe(HAd)3 and Fe(HDr)3. In these formulas, HAd and HDr stand for adriamycin and daunorubicin in which the 1,4-dihydroxy anthraquinone moiety is half-deprotonated. Both complexes are six-membered chelates. The stability constant is beta = (2.5 +/- 0.5) X 10(28) for both complexes. Interaction with DNA has been studied showing that, despite strong coordination to Fe(III), anthracyclines are able to intercalate between DNA bases pairs, releasing the metal. These complexes display antitumor activity against P 388 leukemia that compares with that of the free drug. Fe(HAd)3, unlike adriamycin, does not catalyze the flow of electrons from NADH to molecular oxygen through NADH dehydrogenase. Moreover, it is shown that the triferric adriamycin compound so called "quelamycin" is in fact a mixture of Fe(HAd)3 and polymeric ferric hydroxide. PMID- 2983754 TI - Structures of manganese(II) complexes with ATP, ADP, and phosphocreatine in the reactive central complexes with creatine kinase: electron paramagnetic resonance studies with oxygen-17-labeled ligands. AB - Coordination of Mn(II) to the phosphate groups of the substrates and products in the central complexes of the creatine kinase reaction mixture has been investigated by electron paramagnetic resonance (EPR) spectroscopy with regiospecifically 17O-labeled substrates. The EPR pattern for the equilibrium mixture is a superposition of spectra for the two central complexes, and this pattern differs from those observed for the ternary enzyme-Mn(II)-nucleotide complexes and from that for the dead-end complex enzyme-Mn(II)ADP-creatine. In order to identify those signals that are associated with each of the central complexes of the equilibrium mixture, spectra were obtained for a complex of enzyme, Mn(II)ATP, and a nonreactive analogue of creatine, 1-(carboxymethyl)-2 iminoimidazolidin-4-one, which is a newly synthesized competitive inhibitor. This inhibitor permits an unobstructed view of the EPR spectrum for Mn(II)ATP in the closed conformation of the active site. The EPR spectrum for this nonreactive complex with Mn(II)ATP matches one subset of signals in the spectrum for the equilibrium mixture, i.e., those due to the enzyme-Mn(II)-ATP-creatine complex. Chemical quenching of the samples followed by chromatographic assays for both ATP and ADP indicates that the enzyme-Mn(II)ADP-phosphocreatine and the enzyme Mn(II)ATP-creatine complexes are present in a ratio of approximately 0.7 to 1. A similar value for the equilibrium constant for enzyme-bound substrates is obtained directly from the EPR spectrum for the equilibrium mixture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983755 TI - Labeling of the glycoprotein subunit of (Na,K)ATPase with fluorescent probes. AB - Sodium plus potassium activated adenosinetriphosphatase [(Na,K)ATPase] is composed of a catalytic subunit (alpha) and a glycoprotein subunit (beta) of unknown function. A method has been developed to label the beta subunit of purified dog kidney (Na,K)ATPase with fluorescent probes. The method consists of oxidation of beta-subunit oligosaccharides, reaction of the resulting aldehydes with fluorescent hydrazides, and reduction of the hydrazones and unreacted aldehydes with NaBH4. Two oxidation methods were compared. Simultaneous treatment with neuraminidase and galactose oxidase did not inhibit significantly (Na,K)ATPase activity and allowed insertion of up to 11 mol of probe per mol of beta. In contrast, oxidation of (Na,K)ATPase oligosaccharides with periodate resulted in 50-80% inhibition of the (Na,K)ATPase activity with low or undetectable labeling. Eleven commercial probes and two novel hydrazides were tested for labeling of (Na,K)ATPase treated with galactose oxidase and neuraminidase. Eight probes did not label (Na,-K)ATPase but labeled red cell ghosts oxidized with periodate. Four probes labeled beta specifically but either adsorbed to the membrane tightly, or cross-linked the beta subunits, or formed unstable adducts. Lucifer yellow CH labeled beta specifically without membrane adsorption. Labeling stoichiometries from 1 to 11 mol of lucifer yellow CH per mol of beta were obtained without inhibition of (Na,K)ATPase activity and without significant alteration of the anthroylouabain binding capacity or its association and dissociation kinetics. Anthroylouabain specifically bound to the lucifer labeled (Na,K)ATPase had a decreased quantum yield, probably due to resonance energy transfer. This suggests that the sites of lucifer attachment on beta are within energy transfer distance from the cardiac glycoside site on alpha.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983756 TI - An electron transfer dependent membrane potential in chromaffin-vesicle ghosts. AB - Adrenal medullary chromaffin-vesicle membranes contain a transmembrane electron carrier that may provide reducing equivalents for intravesicular dopamine beta hydroxylase in vivo. This electron transfer system can generate a membrane potential (inside positive) across resealed chromaffin-vesicle membranes (ghosts) by passing electrons from an internal electron donor to an external electron acceptor. Both ascorbic acid and isoascorbic acid are suitable electron donors. As an electron acceptor, ferricyanide elicits a transient increase in membrane potential at physiological temperatures. A stable membrane potential can be produced by coupling the chromaffin-vesicle electron-transfer system to cytochrome oxidase by using cytochrome c. The membrane potential is generated by transferring electrons from the internal electron donor to cytochrome c. Cytochrome c is then reoxidized by cytochrome oxidase. In this coupled system, the rate of electron transfer can be measured as the rate of oxygen consumption. The chromaffin-vesicle electron-transfer system reduces cytochrome c relatively slowly, but the rate is greatly accelerated by low concentrations of ferrocyanide. Accordingly, stable electron transfer dependent membrane potentials require cytochrome c, oxygen, and ferrocyanide. They are abolished by the cytochrome oxidase inhibitor cyanide. This membrane potential drives reserpine sensitive norepinephrine transport, confirming the location of the electron transfer system in the chromaffin-vesicle membrane. This also demonstrates the potential usefulness of the electron transfer driven membrane potential for studying energy-linked processes in this membrane. PMID- 2983757 TI - Effect of hormones on transcription of the gene for cytosolic phosphoenolpyruvate carboxykinase (GTP) in rat kidney. AB - The effect of hormones on the transcription rate of cytosolic phosphoenolpyruvate carboxykinase and level of mRNA for this enzyme in the rat kidney has been investigated. In renal nuclei isolated from rats given dibutyryladenosine cyclic 3',5'-phosphate (Bt2cAMP) or 8-bromoadenosine cyclic 3',5'-phosphate (8-Br-cAMP), [32P]UMP incorporation into hybridizable phosphoenolpyruvate carboxykinase mRNA increased severalfold within 1 h. Changes in the concentration of cytosolic phosphoenolpyruvate carboxykinase mRNA, measured by hybridization of [32P]cDNA to poly(A)+ mRNA, paralleled alterations in the transcription rate. Dexamethasone treatment of adrenalectomized rats increased the transcription rate and the level of phosphoenolpyruvate carboxykinase mRNA 3-4-fold after 4 h. Both parameters then declined to control values by 8 h. When dexamethasone (5 mg/kg) and Bt2cAMP (25 mg/kg) were given together, the rate of phosphoenolpyruvate carboxykinase RNA synthesis and the level of cytosolic mRNA were not increased more than those with either drug alone. Transcription of the gene for renal phosphoenolpyruvate carboxykinase was not affected by diabetes or glucose refeeding but was increased 2-fold after 24 h of starvation and reduced by bicarbonate feeding after 2 h. We conclude that glucocorticoids and cAMP change the rate of transcription of the phosphoenolpyruvate carboxykinase gene in rat kidney, leading to changes of similar magnitude in mRNA level and, hence, enzyme activity. The results presented here and in previous work [Lamers, W., Hanson, R. W., & Meisner, H. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 5137] indicate that the transcription rate of the gene for phosphoenolpyruvate carboxykinase in liver and kidney responds to hormones in a tissue-specific manner. PMID- 2983758 TI - Differential effects of parathyroid hormone responsive cultured human cells on biological activity of parathyroid hormone and parathyroid hormone inhibitory analogues. AB - We have employed parathyroid hormone (PTH) responsive human cells cultured from dermis or giant cell tumors of bone (GT) to evaluate the biological properties of a newly developed in vivo PTH inhibitor, [Tyr34]bPTH-(7-34)-amide (PTH-Inh). Short periods of incubation of cells from dermis or GT with maximal stimulatory concentrations of PTH in the presence of increasing concentrations of PTH-Inh resulted in a dose-dependent inhibition of the adenosine cyclic 3',5'-phosphate (cAMP) response (Ki = 3 X 10(-7) M and 4.2 X 10(-7) M for GT and dermal cells, respectively). In both cell cultures, PTH-Inh alone did not increase cAMP levels, and in desensitization experiments, preincubation with PTH-Inh alone did not desensitize cells to PTH. Hence, the analogue displayed no agonist properties. Unexpectedly, when PTH-Inh was incubated with dermal cells in the presence of PTH, the PTH-Inh failed to block desensitization, suggesting a loss of biological effectiveness of the inhibitor. When medium containing PTH-Inh alone was removed from dermal cells and tested for inhibition of the acute PTH response in untreated cells, there was apparent loss of inhibitory efficacy (t1/2 = 20 h). In contrast, incubation of native PTH or bPTH-(1-34) with cells did not affect the biological activity of these ligands. Unlike the dermal cells, the PTH-Inh did block desensitization to PTH in GT, and there was no loss of inhibitor efficacy when medium containing PTH-Inh was incubated with GT (48 h) and then tested in untreated cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2983759 TI - Nuclear magnetic resonance determination of metal-proton distances in a synthetic calcium binding site of rabbit skeletal troponin C. AB - The binding of gadolinium to a synthetic peptide of 13 amino acid residues representing the calcium binding loop of site 3 of rabbit skeletal troponin C [AcSTnC(103-115)amide] has been studied by using proton nuclear magnetic resonance (1H NMR) spectroscopy. In particular, the proton line broadening and enhanced spin-lattice relaxation have been used to determine proton-metal ion distances for several assigned nuclei in the peptide-metal ion complex. These distances have been used in conjunction with other constraints and a distance algorithm procedure to demonstrate that the structure of the peptide-metal complex as shown by 1H NMR is consistent with the structure of the EF calcium binding loop in the X-ray structure of parvalbumin but that the available 1H NMR distances do not uniquely define the solution structure. PMID- 2983760 TI - Laser light-scattering characterization of mitochondrial complex III-Triton X-100 phospholipid mixed micelles. AB - Bovine-heart mitochondrial complex III was purified in the presence of Triton X 100, and the size and shape of the resulting protein-surfactant-phospholipid mixed micelles were investigated by laser light-scattering. The protein appears to be present in the form of a dimer, irrespective of temperature (between 25 and 40 degrees C) and protein concentration (between 0.5 and 5 mg/ml). The molecular weight of the micelle increases with temperature from 600 000 (25 degrees C) to 692 000 (40 degrees C). The variation of the solvent second virial coefficient in this temperature range suggests that, with increasing temperature, some of the free surfactant molecules become integrated in the mixed micelles. The average quadratic radius of gyration of these is of 42 +/- 5 nm, corresponding in our case to an ellipsoidal shape. PMID- 2983761 TI - Two modes of irreversible inactivation of the mitochondrial electron-transfer system by tetradecanoic acid. AB - Bovine heart submitochondrial particles were incubated for 2-6 h at 37 degrees C with various concentrations of tetradecanoic acid, and the effects on the activities, the total acid-labile sulphide content and EPR spectra of the electron transfer system were studied. Two distinct time-dependent processes of the slow irreversible inactivation of the electron-transfer system were found. They differ in the concentration of tetradecanoic acid required. The more specific effect, induced by 100-400 nmol tetradecanoic acid per mg protein, consists of a selective blockage of electron transfer between the Fe-S clusters of the NADH dehydrogenase and ubiquinone, without damage to any of the Fe-S clusters. Higher concentrations of tetradecanoic acid caused gradual destruction of all Fe-S clusters of NADH dehydrogenase and of the 3-Fe cluster of succinate dehydrogenase, leading to complete inactivation of both NADH and succinate oxidation. PMID- 2983762 TI - Proton-coupled transport of glycylglycine in rabbit renal brush-border membrane vesicles. AB - Transport of glycylglycine into rabbit renal brush-border membrane vesicles was found to be Na+-independent, H+ gradient-dependent and electrogenic. Marked overshoot uptake of the dipeptide was observed when an inward-directed proton gradient and inside-negative potential difference were imposed simultaneously across the vesicular membranes. Saturable depolarization of vesicular membranes could be demonstrated with glycylglycine by use of a fluorescent cyanine dye, di S-C3(5). The results indicate that glycylglycine is contransported with H+ across the membranes. PMID- 2983763 TI - Latency studies on rat liver microsomal glucose-6-phosphatase. Correlation of membrane modification and solubilization by Triton X-114 with the enzymatic activity. AB - Interrelationships between the catalytic properties of glucose-6-phosphatase and the membrane structure of rat liver microsomes were investigated. Membrane modification and solubilization employing the nonionic surfactant Triton X-114 were standardized and analysed by ultracentrifugation, surface tension- and turbidity measurements. The effect of Triton X-114 on the glucose-6-phosphatase activity was studied systematically and the whole magnitude of time- and temperature-dependent inactivation of this enzyme has been demonstrated. The results show that the activity measured is always a resultant of two processes, the beginning of inactivation and the release of latency. Maximal activation of about 600% (83% of apparent latency) was obtained at 0 degree C. A correlation between membrane modification and solubilization and the conditions under preincubation and test incubation reveals that studies on detergent-disrupted microsomes are performed on structures reassembled from solubilizates and this implies a modified microenvironment in the reconstitutes. Kinetic analyses suggest interrelationships between Triton X-114 and the permeability barrier of the glucose-6-phosphatase system. At 0 degree C 2-propanol and ethanol are more potent tools for membrane modification than Triton X-114 and release 88% and 86% latent activity corresponding to an activation of the glucose-6-phosphatase of about 850% and 700%, respectively. These observations suggest that detergent treatment of microsomes could not preserve the functional integrity of the glucose-6-phosphate phosphohydrolase, which is one dogma of the substrate transport hypothesis developed by Arion and his co-workers (Arion, W.J., et al. (1975) Mol. Cell. Biochem. 6, 75-83). PMID- 2983764 TI - Sequential onset of permeability changes in mouse ascites cells induced by Sendai virus. AB - The addition of haemolytic Sendai virus to cells induces membrane changes in the following sequence: (i) Increased permeability to ions, (ii) increased permeability to low molecular weight metabolites, (iii) increased permeability to proteins. The consequences of an increased permeability to ions are: (a) alteration of membrane potential, (b) net changes in intracellular cations and (c) cell swelling, in that order. Depending on virus: cell ratio, Ca2+ concentration and temperature, it is possible to observe ion leakage without metabolite or protein leakage, and ion and metabolite leakage without protein leakage. A model for the induction of permeability changes is presented. PMID- 2983765 TI - Canine neutrophil plasma membrane markers. AB - The purpose of this investigation was to determine which enzyme activities are true canine neutrophil plasma membrane markers. Three enzymes thought to be present on plasma membranes were chosen for study: 5'-nucleotidase, magnesium dependent adenosine triphosphatase (Mg2+-ATPase), and leucine aminopeptidase. Both 5'-nucleotidase and Mg2+-ATPase were found to be ectoenzymes in the canine neutrophil but additional Mg2+-ATPase activity was located intracellularly. An endogenous inhibitor of 5'-nucleotidase was found in the cytosol of canine neutrophils. The specific 5'-nucleotidase inhibitor, adenosine 5'-[alpha, beta methylene] diphosphate also inhibited the canine enzyme in intact cells. Leucine aminopeptidase was located solely in the myeloperoxidase-containing granules of the canine neutrophil. Plasma membrane, as identified by the presence of Mg2+ ATPase and 5'-nucleotidase activities, was separated from other cell organelles by Percoll-density gradient centrifugation of a 10 000 X g supernatant of nitrogen cavitated neutrophils. PMID- 2983766 TI - Active sodium transport in basolateral plasma membrane vesicles from rat kidney proximal tubular cells. AB - Inside-out vesicles prepared with basolateral plasma membranes from rat kidney proximal tubular cells can accumulate Na+ actively in two ways. Mode 1, which is K+-independent, is ouabain-insensitive and is inhibited by furosemide and mode 2, which is K+-dependent, is inhibited by ouabain and is insensitive to furosemide. The presence of Mg2+ and ATP in the incubation medium is essential for both modes of Na+ uptake to proceed and in both cases, the nucleotide is hydrolyzed during the process. These results are consistent with the idea of the existence, in these membranes, of two Na+ pumps: one, which can work in the absence of K+ (Na+ pump) and another, which needs K+ to work (Na+ + K+ pump). PMID- 2983767 TI - Lipid-protein interactions in frog rod outer segment disc membranes. Characterization by spin labels. AB - Freely-diffusing phospholipid spin labels have been employed to study rhodopsin lipid interactions in frog rod outer segment disc membranes. Examination of the ESR spectra leads us to the conclusion that there are two motionally distinguishable populations of lipid existing in frog rod outer segment membranes over a wide physiological temperature range. Each of the spin probes used shows a two-component electron spin resonance (ESR) spectrum, one component of which is motionally restricted on the ESR timescale, and represents between 33 and 40% of the total integrated spectral intensity. The second spectral component which accounts for the remainder of the spectral intensity possesses a lineshape characteristic of anisotropic motion in a lipid bilayer, very similar in shape to that observed from the same spin labels in dispersions of whole extracted frog rod outer segment lipid. The motionally restricted spectral component is attributed to those spin labels in contact with the surface of rhodospin, while the major component is believed to originate from spin labels in the fluid lipid bilayer region of the membranes. Calculations indicate that the motionally restricted lipid is sufficient to cover the protein surface. This population of lipids is shown here and elsewhere (Watts, A., Volotovski, I.D. and Marsh, D. (1979) Biochemistry 18, 5006-5013) to be by no means rigidly immobilized, having motion in the 20 ns time regime as opposed to motions in the one nanosecond time regime found in the fluid bilayer. Little selectivity for the motionally restricted population is observed between the different spin-labelled phospholipid classes nor with a spin-labelled fatty acid or sterol. PMID- 2983768 TI - The effect of fructose 2,6-bisphosphate on muscle fructose-1,6-bisphosphatase activity. AB - Rat and rabbit muscle fructose 1,6-bisphosphatase (D-fructose-1,6-bisphosphate 1 phosphohydrolase, EC 3.1.3.11) are inhibited by fructose 2,6-bisphosphate. In contrast with the liver isozyme, the inhibition of muscle fructose-1,6 bisphosphatase by fructose 2,6-bisphosphate is not synergistic with that of AMP. Activation of fructose-1,6-bisphosphatase by fructose 2,6-bisphosphate has been observed at high concentrations of substrate. An attempt is made to correlate changes in concentrations of hexose monophosphate, fructose 1,6-bisphosphate and fructose 2,6-bisphosphate with changes in fluxes through 6-phosphofructokinase and fructose-1,6-bisphosphatase in isolated epitrochlearis muscle challenged with insulin and adrenaline. PMID- 2983769 TI - Thermal denaturation of rat pulmonary and testicular angiotensin-converting enzyme isozymes. Effects of chelators and CoCl2. AB - In an attempt to assess the biochemical consequences resulting from structural differences between rat pulmonary and testicular angiotensin-converting enzyme, the thermal stability of crude and purified preparations of each enzyme was compared. Structural heterology was verified by molecular weight determinations and by peptide mapping after limited proteolysis with Staphylococcus V8 proteinase. Thermal stability was monitored by changes in catalytic activity following incubations at 55 degrees C in the presence of chelators and CoCl2. Purified pulmonary angiotensin-converting enzyme was more sensitive to inhibition by the chelators EDTA and 1,10-phenanthroline and by the site-directed inhibitor captopril than was the testicular isozyme. Although the pulmonary holoenzyme was unaffected by cobalt, the testicular holoenzyme was inhibited by cobalt in a concentration-dependent manner. Crude pulmonary angiotensin-converting enzyme was significantly more resistant to thermal denaturation than its crude testicular counterpart. The differences in the thermal lability of each isozyme were still present in purified preparations, although the purified enzymes appeared to be more thermally stable than their crude counterparts. Both chelators and cobalt markedly potentiated the thermal denaturation of each isozyme. These data suggest that the structural heterology of the pulmonary and testicular isozymes may affect the interaction of zinc with the respective enzymes and that zinc may contribute to the structural integrity and thermal stability of angiotensin converting enzyme in each tissue. PMID- 2983770 TI - Improved procedures for the synthesis of phosphomevalonate and for the assay and purification of pig liver phosphomevalonate kinase. AB - An improved procedure for the synthesis of phosphomevalonate using excess free ATP4-, and phenyl agarose to remove contaminating nucleotides, is described. A high-voltage electrophoresis assay, which separates phosphomevalonate from mevalonate 5-diphosphate at pH 3.5, was developed for the assay of phosphomevalonate kinase (ATP:5-phosphomevalonate phosphotransferase, EC 2.7.4.2). High-voltage electrophoresis, at pH 5, could also be used for the separation of mevalonate 5-diphosphate from isopentenyl diphosphate. An alternative method for the purification of phosphomevalonate kinase from pig liver was also developed. The high-voltage electrophoresis assay was used to reassess the metal ion and nucleotide specificity of the pig liver phosphomevalonate kinase. ATP could be partially replaced by ITP and GTP and poorly by CTP and UTP. Apparent activation of the enzyme by free ATP4- was observed as found for mevalonate kinase (C.S. Lee and W.J. O'Sullivan (1983) Biochim. Biophys. Acta 747, 215-224). PMID- 2983772 TI - Possible involvement of two mechanisms of signal transduction in alpha 1 adrenergic action. Selective effect of cycloheximide. AB - We have previously suggested that the effects of alpha 1-adrenergic agents on hepatocyte metabolism involve two pathways: (a) a calcium-independent, insulin sensitive process which is modulated by glucocorticoids; and (b) a calcium dependent, insulin-insensitive process which is modulated by thyroid hormones. Cycloheximide stimulated ureogenesis through a prazosin-sensitive mechanism in liver cells (alpha 1-adrenergic). The effect of cycloheximide was insulin insensitive and calcium-dependent. Furthermore, a clear effect of cycloheximide was observed in hepatocytes obtained from adrenalectomized animals, whereas no effect was observed in cell from hypothyroid rats. The effects of epinephrine and cycloheximide were blocked by phorbol esters in all the conditions tested. Binding competition experiments indicated that cycloheximide interacts with only a fraction of the alpha 1-adrenergic sites labeled with [3H]prazosin. It is suggested that cycloheximide activates preferentially one of the pathways involved in the alpha 1-adrenergic action in liver cells. PMID- 2983771 TI - The use of 36Cl- to measure cell plasma membrane potential in isolated hepatocytes--effects of cyclic AMP and bicarbonate ions. AB - The plasma membrane potential of hepatocytes was calculated from the distribution of 36Cl-. The potential observed under several conditions was equivalent to that previously measured using microelectrodes in perfused liver. Dibutyryl cAMP increased the membrane potential. Replacement of bicarbonate ions by morpholinosulphonate decreased the potential and reduced the effect of cAMP. The effect of both bicarbonate and cAMP was abolished by ouabain. Both bicarbonate and cAMP stimulated the activity of the (Na+ + K+)-ATPase as measured by ouabain inhibitable 86Rb+ uptake. It is suggested that the stimulation of alanine transport by these effectors is mediated by an increase in cell membrane potential via stimulation of the (Na+ + K+)-ATPase. PMID- 2983773 TI - 5'-Nucleotidase activity of isolated mature rat cardiac myocytes. AB - Specific location of 5'-nucleotidase in the heart has been uncertain, some authors citing evidence for an exclusively non-myocyte location, while other data point to the existence of cytoplasmic and membrane-bound fractions. Single myocytes isolated from mature rat heart, and free of endothelial or interstitial cells, have been used to establish that muscle cells of the myocardium are rich in 5'-nucleotidase, exhibiting activity sufficient to account for the total myocardial content of this enzyme. All 5'-nucleotidase is accessible to extracellular AMP. Inhibitors of 5'-nucleotidase and adenosine transport have been used to establish that only the adenosine component of adenine nucleotides is taken up by myocytes, but hydrolysis of AMP by 5'-nucleotidase does not commit the adenosine formed to transport across the sarcolemmal membrane. Myocytes also have ecto-phosphatases which hydrolyse ADP and ATP. PMID- 2983774 TI - Transmembrane modulation of the concanavalin A inhibition of 5'-nucleotidase is not due to a direct association of the enzyme with the cytoskeleton. AB - The inhibition of the cell surface enzyme 5'-nucleotidase by concanavalin A is being studied as a model for understanding transmembrane modulation of cell surface functions. Nucleotidase of 13762 MAT-C1 ascites rat mammary adenocarcinoma cells is inhibited by concanavalin A in a noncooperative process. When cells are treated with the cytoplasmic effectors cytochalasins, colchicine, energy poisons, calcium plus ionophore or hypotonic buffers, the concanavalin A inhibition of the enzyme becomes cooperative. 5'-Nucleotidase of isolated MAT-C1 microvilli is also inhibited by concanavalin A in a noncooperative process; however, treatment of the microvilli with the same cytoplasmic effectors does not induce cooperativity. Since previous studies in several systems have suggested an association of nucleotidase with actin-containing microfilaments or the cell cytoskeleton, one explanation for the cooperativity changes is that they result from a change in the association of the enzyme with the cytoskeleton. However, Triton X-100 extractability of nucleotidase is the same for MAT-C1 cells exhibiting cooperative or noncooperative concanavalin A inhibition. Moreover, enzyme from cells exhibiting cooperative inhibition can be extracted into the zwitterionic detergent Zwittergent in a cooperative form, while enzyme exhibiting noncooperative behavior can be extracted into Zwittergent in a noncooperative form. Gel filtration and rate-zonal sucrose density gradient centrifugation showed little discernible size or sedimentation difference between enzyme samples exhibiting noncooperative and cooperative inhibition. These results indicate that changes in the cooperativity of the concanavalin A inhibition of nucleotidase are not a result of changes in the association of the enzyme with the cytoskeleton. These studies emphasize the caution which must be exercised in interpreting the effects of cytoskeletal perturbants on cell surface functions. PMID- 2983775 TI - Vanadate, tungstate and molybdate activate rod outer segment phosphodiesterase in the dark. AB - Anionic activation of rod outer segment phosphodiesterase by vanadate, molybdate and tungstate is demonstrated. Comparisons are made to adenylate cyclase, which is known to be activated by vanadate and molybdate but not by tungstate. In view of the differences in anionic activation between these two important enzymatic regulators of intracellular cyclic nucleotide metabolism, it is possible that tungstate can be used as a selective probe for the effects of phosphodiesterase activity in photoreceptors and other cells. The known electrophysiological stimulation of Limulus photoreceptors by these anions is also interpreted in light of our results. If anionic production of quantum bumps in Limulus photoreceptors is mediated by changes in cyclic nucleotides, then the electrophysiological response of Limulus photoreceptors to tungstate may indicate a role for phosphodiesterase rather than adenylate cyclase in mediating light induced cyclic nucleotide alterations in this cell. PMID- 2983776 TI - [Mathematical model of the dynamics of transport of inert gases in the microcirculatory system]. AB - A mathematical model imitating transport of inert gases in the system of microcirculation under increased pressures was constructed. It has been shown that saturation of microareas nucleus of the brain cortex of average dimensions proceeds in about 90 sec. Effect of the blood flow velocity, gases tension in arterial blood and density of the capillary net on the dynamics of mass transfer of gases in a tissue was investigated. PMID- 2983777 TI - [Segmental motility of the COOH-terminal fragment of immunoglobulin G heavy peptide chains in a solution]. AB - The rotational correlation time of pFc' fragment of IgG1 determined by spin-label method was found to be equal to 5.2 nsec. This value points to the significant segmental mobility of Ch3 domains built fragment as well as to the structural lability of Ch3 domains themselves. PMID- 2983778 TI - [The lack of the effect of a strong constant magnetic field on isolated membrane preparations of Na,K-dependent ATPase]. AB - Effect of constant magnetic field (CMF) with induction 10 T on membrane preparations of Na,K-dependent ATPase of bovine brain (lipoproteid vesicules with 300-500 A diameter) were studied. No CMF effect on the activity of Na,K-dependent ATPase was observed under different experimental conditions (three temperature points 15, 20 and 37 degrees C and great variation of Na+,K+ concentrations ratio). CMF also produced no effect on the preparations of Na,K-dependent ATPase immobilized by adsorption on millipore filters. PMID- 2983779 TI - [Interaction of spin-labeled methacyne analog with butyrylcholinesterase]. AB - Interaction between spin-labeled methacyne (I) and butyrylcholinesterase (BChE) was studied by ESR and enzyme kinetic methods. The compound (I) was shown to be a competitive reversible inhibitor, the value of Ki appeared to be 1.3 X 10(-5) M. Insertion of nitroxyl fragment in the methacyne molecule results in a two-fold increase of its inhibitory activity. The ESR spectrum of the enzyme-inhibitor complex was registered. This complex dissociates under the action of eserine, tetramethylammonium and hexamethonium. Scatchard plot reveals two different types of binding sites with Kdiss values 1.5 X 10(-5) M and 2.6 X 10(-4) M. One type of binding sites is identified as the enzyme active centre. The restricted motion of (I) in complex with BChE proves the assumption that the enzyme active centre is located in the split of macromolecule surface. PMID- 2983780 TI - [Study of the structure of erythrocyte membrane in spontaneous genetic hypertension using the spin-label method]. AB - An analysis of ESR spectra of maleimid spin-labeled erythrocyte membranes of spontaneously hypertensive rats of SHR line and normotensive rats of the control line WKY showed differences in the structure of membrane proteins in the norm and pathology. These differences were compared with the differences between the erythrocyte membranes of SHR and WKY, found earlier by fluorescent probe method. An important role of membrane peripheric proteins in the appearance of the above differences in suggested. PMID- 2983781 TI - [The nature of thyroid hormone receptors. The role of serum thyroxine binding prealbumin in the realization of the hormonal effect]. AB - Data from determination of molecular weight and competitive displacement suggest that T3 and T4 are bound to the same protein in chromatin. It was shown that the antigenic determinants of T3 and T4 for the chromatin-binding protein coincide with those for blood serum thyroxine-binding prealbumin (TBPA). It was found also that the binding either to T3 and T4 decreases proportionally to the amount of the TBPA removed from the subcellular fractions. It may thus be concluded that blood serum TBPA is responsible for the binding to T3 and T4 as well as for the realization of the hormonal response. PMID- 2983782 TI - [Changes in the cAMP-dependent phosphorylation system of myocardial sarcoplasmic reticulum components during circulatory hypoxia]. AB - The dissociation of the holoenzyme of the cAMP-dependent protein kinase was induced by the adrenaline-stimulated endogenous cAMP synthesis. This dissociation was impaired in a hypoxic muscle. The elevated proteolytic activity in the hypoxic area was found to be associated with the reduction of the 33P incorporation into phospholamban in the presence of 10(-6) M cAMP. No changes were observed when protein kinase was added to the incubation mixture. A correlation between the degree of phospholamban phosphorylation and its susceptibility to proteolysis was demonstrated. PMID- 2983783 TI - [Kinetics of passive transport of Ca2+ in vesicular preparations of myocardial sarcolemma with inside-out oriented cytoplasm]. AB - Using affinity chromatography on a concanavalin A-Sepharose 4B column, two fractions of rabbit myocardium with oppositely oriented sarcolemmal vesicles have been obtained. Analysis of 45Ca2+ release from the vesicles with inside-out oriented cytoplasm demonstrates that this reaction is biphasic and obeys a pseudo first-order kinetics. The initial rate of Ca2+ release is equal to 0.57 nmol/mg/s. The release of Ca2+ from the vesicles is inhibited via phosphorylation of sarcolemmal proteins by the catalytic subunit of cAMP-dependent protein kinase; the initial rate of this process drops to 0.08 nmol/mg of protein/s. The reaction is also inhibited by Cd2+ greater than Mn2+ greater than Co2+, when the latter are present inside the vesicles. PMID- 2983785 TI - [Effect of phosphatidylglycerol on the incorporation process of cytochrome P-450 in liposomes from dimyristoylphosphatidylcholine]. AB - Using the electron paramagnetic resonance (EPR) method with spin-labeled fatty acids and gel-penetrating chromatography, the effect of phosphatidylglycerol on cytochrome P-450 incorporation into liposomes from dimyristoylphosphatidylcholine was investigated. An addition of phosphatidylglycerol caused an increase in the protein content of the proteoliposomes as well as their accelerated formation at temperatures below and above the liposome phase transition temperature (Ts). The dependence of the proteoliposome formation rate on the phosphatidylglycerol content in the liposome mixture is described by complex kinetics with a maximum in the presence of 10 mol.% of the negatively charged phospholipid. The mechanism of proteoliposome formation is discussed in terms of asymmetric distribution and phase state of the phospholipids in the original vesicles. PMID- 2983784 TI - [Use of spin-labelled substrate analogs for a comparative study of microsomal cytochrome P-450 and P-448]. AB - The previously described, iodine-labeled alkylating stable nitroxyl radicals located at different distances between the N-O. group and the iodine atom were used for a comparative study of the structure of microsomal cytochromes P-450 and P-448 active centers. The radicals were shown to change the optical spectra of Fe3+ located in the active site of the enzyme that are similar to those induced by cytochrome P-450 substrates. Some differences in the type of the radicals binding to control, phenobarbital- and 3-methylcholanthrene-induced microsomes were revealed. The alkylating radical substrate analogs covalently bound to microsomal cytochrome P-450 in the vicinity of the active center, resulting in the inhibition of oxidation of type I and II substrates (e. g., aniline and naphthalene). The value of the spectral binding constant (Ks) for naphthalene in the presence of the radical covalently bound to the cytochrome P-450 active center showed a tendency to increase. Using the ESR technique, the interaction between Fe3+ and the radical localized in the active site of cytochrome P-450 was demonstrated. The contribution of Fe3+ to the relaxation of the radicals covalently bound to cytochrome P-450 was evaluated from the values of the spin label ESR spectra saturation curves at 77K. The distances between the N-O. group of these radicals and Fe3+ in the enzyme active center for the three types of microsomes were determined. The data obtained point to structural peculiarities of the active center of cytochrome P-450, depending on the microsomal type. PMID- 2983786 TI - Induction in utero of hepatic glucose-6-phosphatase by fetal hypoinsulinemia. AB - The effect of fetal hypoglycemia and hypoinsulinemia on fetal rat hepatic glucose 6-phosphatase activity was studied. Fetal hypoglycemia and hypoinsulinemia were produced by inducing maternal hyperinsulinemia and hypoglycemia secondary to the exogenous administration of insulin via implantation of osmotically driven minipumps on day 15 of gestation into 15 experimental animals. 13 animals served as sham-operated controls. Cesarean sections were performed on day 20 or 21 of gestation under pentobarbital anesthesia. Liver glucose-6-phosphatase activity was increased in the hypoinsulinemic fetuses. In contrast, the hyperinsulinemic mothers had suppressed hepatic glucose-6-phosphatase activity. Hypoinsulinemia would appear to be the primary stimulus for enhanced fetal glucose-6-phosphatase in this model. PMID- 2983787 TI - Human plasma inhibitors of platelet serotonin uptake and imipramine receptor binding: extraction and heterogeneity. AB - Human plasma contains a considerable concentration of low molecular weight substances that inhibit, in a dose-dependent manner, both high-affinity imipramine receptor binding and serotonin uptake in platelets. Incubation of plasma with alumina was used to extract and to partly characterize these imipramine-like inhibitors. The human plasma extract inhibited imipramine binding and serotonin uptake with median inhibitory concentration (IC50) of 0.18 +/- 0.1 and 0.36 +/- 0.15 mg/ml, respectively. Imipramine-like activity of the extract was markedly degraded by carboxypeptidase B and leucine aminopeptidase, but was resistant to neurominidase and phospholipases A2, C, and D. The elution profile of the extract after gel chromatography on Bio-Gel P-2 showed two major peaks of serotonin uptake and imipramine binding inhibition and three additional peaks of serotonin uptake inhibitory activity that did not have a significant effect on imipramine binding. The possible mechanism of pharmacological action of the imipramine-like inhibitors and their relation to development of affective illnesses remain to be clarified. PMID- 2983788 TI - ACTH, cortisol, and corticosterone output after ovine corticotropin-releasing factor challenge during depression and after recovery. AB - Synthetic ovine corticotropin releasing factor (o-CRF) was administered as an intravenous bolus (100 micrograms) to eight patients suffering from a major depressive disorder, endogenous subtype. All patients showed inadequately suppressed cortisol levels after 1 mg dexamethasone. After clinical remission and normalized dexamethasone responses, these patients were reinvestigated with o-CRF stimulation. The mean adrenocorticotropic hormone (ACTH) release from the pituitary corticotroph cells was indiscriminate at both test sessions. Cortisol and corticosterone output after o-CRF tended to be higher during depression than after recovery. The o-CRF-induced increments observed with corticosterone were more marked in comparison with cortisol. Within the limitations of the current protocol, our preliminary data lend support to the view that an increased pituitary ACTH reserve or adrenocortical steroid reserve is not likely to be responsible for the defective pituitary-adrenal regulation in some dexamethasone resistant depressives. PMID- 2983789 TI - Alteration of platelet [3H]imipramine binding in mildly depressed patients correlates with disease severity. PMID- 2983790 TI - Solubility of lithium salts in organic solvents. PMID- 2983791 TI - Disassembly from both ends of thick filaments in rabbit skeletal muscle fibers. An optical diffraction study. AB - We show in this paper that the change of the internal structure of a sarcomere in a rabbit glycerinated psoas muscle fiber can be examined by analyzing the intensity change of the first- and the second-order optical diffraction lines. A unit-cell (sarcomere)-structure model has been applied to the estimation of the length of thick filaments in a muscle fiber while they undergo dissociation. The optical factors, except for the unit-cell-structure factor, hardly changed during the dissociation of the filaments. Our results show that thick filaments dissociate from both ends on increasing the KCl concentration in the presence of 10 mM pyrophosphate and 5 mM MgCl2. Micromolar concentrations of Ca2+ suppressed to some extent the dissociation of thick filaments. The disassembly of thick filaments occurred at higher KCl concentrations in the absence of pyrophosphate. There was a correlation between the stability of the thick filament structure and cross-bridge formation, which was induced either by the addition of micromolar concentrations of Ca2+ in the presence of Mg-pyrophosphate or by removal of Mg pyrophosphate. PMID- 2983792 TI - Disassembly kinetics of thick filaments in rabbit skeletal muscle fibers. Effects of ionic strength, Ca2+ concentration, pH, temperature, and cross-bridges on the stability of thick filament structure. AB - The kinetics of dissociation from both ends of thick filaments in a muscle fiber was investigated by an optical diffraction method. The dissociation velocity of thick filaments at a sarcomere length of 2.75 microns increased with increasing the KCl concentration (from 60 mM to 0.5 M), increasing the pH value (from 6.2 to 8.0) or decreasing the temperature (from 25 to 5 degrees C) in the presence of 10 mM pyrophosphate and 5 mM MgCl2. Micromolar concentrations of Ca2+ suppressed the dissociation velocity markedly at shorter sarcomere lengths. The dissociation velocity, v, decreased as thick filaments became shorter, and v = -db/dt = vo exp (alpha b), where b is the length of the thick filament at time t and vo and alpha are constants. The vo value was largely dependent on the KCl concentration but the alpha value was not. The stiffness of a muscle fiber decreased nearly in proportion to the decrease of overlap between thick and thin filaments induced by the dissociation of thick filaments. This indicates that cross-bridges are uniformly distributed and contribute independently to the stiffness of a muscle fiber during the dissociation of thick filaments. PMID- 2983793 TI - Electrochemical and electron spin resonance studies of actinomycin D and other phenoxazones. AB - Electrochemical studies on actinomycin D (1) and two analogs, 2-amino-3 phenoxazone (2) and 1,2,4-trichloro-7-nitrophenoxazone (3) were analyzed by polarography and ESR spectroscopy. The polarograms of the three compounds in acetonitrile all show two reduction waves. ESR experiments confirm that the first reduction wave corresponds to a one-electron transfer process which produces a phenoxazone free radical anion and the second wave corresponds to a subsequent one-electron transfer producing a diamagnetic dianion. Substitution with electron withdrawing groups such as NO2 (at C-7) and chloro (at C-1, C-2 and C-4)3 facilitated the reduction of the phenoxazone ring system to a free radical (i.e., half-wave potentials; 1, -0.815 V; 2, -0.920 V; 3, -0.135 V). It was found, by computer simulation of the ESR spectra, that the spin density in the electrochemically generated free radicals from 1, 2 and 3 was preferentially located in the benzenoid ring and at the N-10 nitrogen. For radicals obtained from 1 and 2, only a small residual spin density could be detected in the quinoid ring. Since 1 can be metabolized to a free radical in cells, these free radical forms of 1 and its analogs may represent reactive forms of the phenoxazone nucleus. PMID- 2983794 TI - Electron microscopy of the melting of sequenced DNA. PMID- 2983795 TI - Receptor distribution and the endothelial uptake of transcobalamin II in liver cell suspensions. AB - To determine the nature of binding of transcobalamin II (TC-II) to liver cells, we covalently coupled purified holo-TC-II to submicron latex minibeads using glutaraldehyde. Incubation of the probe with liver cell suspensions at 4 degrees C led to its binding by endothelial cells but not by hepatocytes or Kupffer cells, as visualized by scanning electron microscopy. At 37 degrees C, the probe was internalized by the endothelium through a system of coated pits and vesicles as shown by transmission electron microscopy. Inhibition studies by pre incubation with excess native TC-II demonstrated the specificity of binding. Fractionation of these cell suspensions on metrizamide gradients yielded large cell (hepatocyte-rich) and small cell (endothelium-rich) fractions. The binding of the minibead probe occurred again exclusively on endothelial cells in the small cell fraction. 125I-labeled holo-TC-II also bound to the small cell but not to the large cell fraction. Binding was saturable (Ka, 0.225 X 10(9) mol/L-1) and receptor number was calculated to be 1.33 X 10(3) per cell. Time-dependent incubation of 125I-labeled TC-II with the endothelium-rich fraction led to its uptake, reaching a steady-state plateau at 4 degrees C. At 37 degrees C, however, the initial uptake was followed by gradual release of the label into the medium. We conclude that in the liver, holo-TC-II binds initially to endothelium, where it is internalized and is subsequently released probably to the interstitial space. Thus, the endothelium may play a fundamental role in the regulation of the uptake of TC-II by the liver. PMID- 2983796 TI - Absence of a role for superoxide anion, hydrogen peroxide and hydroxyl radical in endothelium-mediated relaxation of rabbit aorta. AB - We tested the hypothesis that the endothelium-dependent relaxation of rabbit thoracic aorta in vitro is mediated by reduced metabolites of oxygen. Helical vascular strips were contracted with either norepinephrine or phenylephrine. Oxygen metabolites, generated by the xanthine oxidase reaction, completely relaxed norepinephrine-induced contractile tone but not tone induced by phenylephrine. A mixture of oxygen metabolite scavengers (superoxide dismutase, catalase and mannitol) eliminated the relaxation induced by the xanthine oxidase products. Acetylcholine caused a dose-dependent and endothelium-dependent relaxation of the strips; this was not inhibited by the presence of the scavengers. We conclude that reduced oxygen metabolites have little direct effect on rabbit aortic smooth muscle in vitro, although they indirectly but specifically relax norepinephrine-induced tone, presumably by oxidation of norepinephrine. Oxygen metabolites do not appear to mediate the endothelium dependent relaxation response of this tissue to acetylcholine. PMID- 2983797 TI - Estrogen receptor separation employing sucrose deuterium oxide gradients generated during sample centrifugation. AB - A quick and easy method of sucrose density gradient analysis is described in which prior preparation of gradients for protein separation is not required. We compare the refractive indices as well as the separation pattern of radioactively labeled proteins with either preformed gradients (prepared by layering or by gradient marker) or gradients that establish themselves during centrifugation. Our results show that an identical pattern of protein separation can be achieved by layering the nuclear extracted estrogen receptor either on top of one single sucrose concentration or on a preformed gradient. Centrifugation times are 40 hr in a swinging bucket type rotor (SW60) or 15 hr in a vertical tube rotor (VTi 65). This method avoids the onerous task of gradient preparation, and the separation itself can be achieved in a 15 hr centrifugation by use of a vertical tube rotor. PMID- 2983798 TI - Three-dimensional atypical structure in intraductal carcinoma differentiating from papilloma and papillomatosis of the breast. AB - The basic architectural pattern of intraductal proliferative lesions of the breast was established by reconstructing serial sections of luminal spaces and interluminal areas of glandular structures. The materials were surgical specimens from twenty patients with intraductal carcinoma, papilloma, papillomatosis, or so called borderline lesion. In papilloma and papillomatosis, the luminal spaces were tubular and interconnected forming a three-dimensional (3-D) network, whereas carcinoma was a porous structure with dispersed lumina. The latter represented 3-D atypical structure in intraductal carcinoma. In borderline lesion the architecture was an intermediate type, with separate lumina partially transformed into tubular shapes. It was also confirmed that the porous structure of carcinoma observed in 3-D reconstructions corresponded to the cribriform pattern seen in 2-D figures, while the network of papilloma produced a complex glandular pattern. A geometric parameter was devised to measure the different 3-D patterns of lumina and their 2-D expressions. It was concluded that the 3-D architectural pattern of intraductal proliferations was sufficiently characteristic to be of diagnostic value in differentiating these diseases. PMID- 2983799 TI - Effect of copper sulphate on respiration, electron transport, and redox potential in the digestive gland of the snail host, Lymnaea luteola. PMID- 2983800 TI - Atypical characteristics of the beta-adrenoceptor mediating cyclic AMP generation and lipolysis in the rat adipocyte. AB - The characteristics of the rat epidydimal adipocyte beta-adrenoceptor have been examined using lipolysis and cyclic AMP accumulation in adipocytes as well as adenylate cyclase activity in fat cell membranes. The pA2 values corrected for binding to bovine serum albumin of the selective antagonists betaxolol (beta 1 selective) and ICI 118.551 (beta 2-selective) against noradrenaline or fenoterol stimulated lipolysis were indicative of an atypical beta-adrenoceptor associated with the lipolytic response. Antagonism of isoprenaline-stimulated cyclic AMP accumulation in whole cells and adenylate cyclase activity in membranes yielded pA2 values to betaxolol, ICI 118.551 and (-)-propranolol, which suggested that the atypical beta-adrenoceptor was coupled to adenylate cyclase. Comparisons of the Ki values obtained in binding studies using [125I]-cyanopindolol with pA2 values obtained in adenylate cyclase experiments suggest that the typical beta 1 receptor identified with radioligand binding studies is not the only receptor site mediating stimulation of adenylate cyclase activity and lipolysis. PMID- 2983801 TI - Two distinct corticotrophin releasing activities of vasopressin. AB - The effect of various corticotrophin releasing factors (CRFs) on the secretion of corticotrophin (ACTH) by segments of rat anterior pituitary tissue has been studied in vitro. ACTH release was stimulated by CRF-41 (5.0 X 10(-9) - 2.0 X 10( 7) M), hypothalamic extracts (0.2 - 1.6 HE ml-1) and arginine vasopressin (AVP, 5.0 X 10(-9) - 8.0 X 10(-7) M). The slopes of the dose-response lines of CRF-41 were greater than those of AVP, less than those of hypothalamic extracts from control animals and resembled those of hypothalamic extracts from Brattleboro rats. Simultaneous addition of AVP (10(-10) M) to the incubation medium enhanced the response to CRF-41 and increased the slope of its dose-response lines. The adrenocorticotrophic response to CRF-41 was enhanced similarly by pretreatment of the tissue with AVP (1.25 X 10(-11) - 2.0 X 10(-10) M) as also was the response to hypothalamic extracts. In contrast, pretreatment of the tissue with CRF-41 (2.3 X 10(-11) - 2.3 X 10(-8) M) depressed the subsequent response to CRF-41 and to hypothalamic extracts. The marked difference between the concentrations of vasopressin required to facilitate maximally the response to CRF-41 and those necessary to evoke ACTH release suggests that the two effects may be mediated by different types of vasopressin receptor. PMID- 2983802 TI - Muscarine and luteinizing hormone releasing hormone attenuate adrenaline induced hyperpolarization in amphibian sympathetic ganglia. AB - The adrenaline-induced hyperpolarization (AdH) and the responses evoked by muscarine and luteinizing hormone releasing hormone (LHRH) were recorded from neurones in amphibian sympathetic ganglia by means of the sucrose gap technique. The amplitude of the AdH was reduced when 'M-channel' closure was promoted by superfusion of LHRH or muscarine. 4-Aminopyridine (4-AP, 1 mM) antagonized the AdH, but not the depolarization evoked by muscarinic agonists. This implies that the channels involved in the electrogenesis of the AdH have different pharmacological properties from 'M-channels' and that the AdH is not generated by the opening of 'M-channels' outside their normal voltage range. Possible explanations for the attenuation of the AdH by muscarine and LHRH might be that (i) intracellular biochemical changes produced by these substances somehow interfere with the generation of the AdH or that (ii) muscarine and LHRH have allosteric interactions with the adrenoceptor mediating the AdH. PMID- 2983803 TI - Cyclic nucleotides and contractility of isolated soleus muscle. AB - The effects of isoprenaline, terbutaline and forskolin were examined on cyclic nucleotide concentrations and contractile responses in guinea-pig isolated soleus muscles. Isoprenaline and terbutaline induced rapid, concentration-related reductions in the tension and degree of fusion of subtetanic contractions of the soleus muscle. These changes were associated with increases (about 2 fold) in the levels of adenosine 3':5'-cyclic monophosphate (cyclic AMP) in the muscle cells. Propranolol competitively inhibited these responses. Forskolin failed to elicit a sympathomimetic response in the soleus muscles despite increasing (by about 20 fold) the intracellular concentration of cyclic AMP. Forskolin also failed to potentiate the effects induced by isoprenaline. The levels of cyclic GMP in the soleus were increased by isoprenaline (about 1.5 fold) and forskolin (about 2.5 fold). Terbutaline was without effect on cyclic GMP levels. These data suggest either that cyclic AMP is not involved as the mediator underlying beta adrenoceptor-induced changes in contractility of slow contracting skeletal muscles or that forskolin does not stimulate the particular adenylate cyclase that leads to appropriate increases in cyclic AMP in those functional compartments associated with modulation of intracellular Ca2+ movements. Cyclic GMP is not involved in modifying changes in contractility of the soleus muscle. PMID- 2983804 TI - The determination of presynaptic KA values of methacholine and pilocarpine and of a presynaptic receptor reserve in the rat perfused heart. AB - Rat isolated perfused hearts with the right sympathetic nerves attached were loaded with [3H]-noradrenaline. The nerves were stimulated with up to 11 trains of 10 pulses at 0.1 Hz. The evoked increases of [3H]-noradrenaline overflow into the perfusate were measured in the presence of cocaine, corticosterone and propranolol. Activation of presynaptic muscarinic receptors by methacholine or pilocarpine inhibited the evoked transmitter release in a reversible and concentration-dependent manner. Preperfusion with phenoxybenzamine (5 microM) for 15 min (followed by a washout of 35 min) changed neither resting nor evoked overflow of [3H]-noradrenaline. The concentration-response curve of methacholine was shifted to the right after exposure of the hearts to phenoxybenzamine (1 microM) without depression of the maximum effect. Pretreatment with phenoxybenzamine (5 microM) reduced the maximum inhibition of release by about 50%. Analysis of the data gave a dissociation constant for the agonist-receptor complex (KA) of 4.0 microM and a receptor reserve of roughly 70%. Half-maximal inhibition of [3H]-noradrenaline release occurred when about 2% of the total receptor population was occupied. Comparison of the concentration-response data for methacholine and pilocarpine revealed a relative efficacy (methacholine/pilocarpine) of 16, a KA of 10 microM for pilocarpine and no receptor reserve for this agonist. The results show that KA values for methacholine and pilocarpine obtained at presynaptic receptors are similar to those obtained at postsynaptic muscarinic receptors. This is in agreement with the idea that muscarinic receptors located on postganglionic adrenergic nerves are not different from those located on effector sites of non-neuronal tissue. PMID- 2983805 TI - Reversal of paralysis in nerve-muscle preparations isolated from animals with hereditary motor endplate disease. AB - Motor endplate disease (med) in the mouse is an hereditary disorder of the skeletal neuromuscular system. Affected animals suffer a 'functional denervation' of skeletal muscle (Duchen & Stefani, 1971). Muscle fibres do not respond to indirect excitation, but motor nerve terminals release transmitter spontaneously. Spontaneous transmitter release can be enhanced by raising [K+]o or by exposing muscles to red-back spider venom and functional transmission following indirect stimulation may be restored by 4-aminopyridine. PMID- 2983806 TI - Fast deactivation of guinea-pig isolated ileum to C5adesArg: a possible cyclic AMP-dependent mechanism. AB - The fast component of deactivation of guinea-pig isolated ileum to the spasmogenic action of the complement peptide C5adesArg was further differentiated from the slow component which had been previously analysed (Damerau et al., 1985a, b). Fast deactivation differs from the slow component in the following characteristics: (a) it is unspecific in that it is also induced by C3a, another complement peptide, (b) it depends on the spasmogenic effect of the peptides, and (c) it does not occur at 16 degrees C. In contrast to the slow component, in which the deactivation is thought to be caused by blockade of C5a receptors, fast deactivation seems to be due to a transient increase of intracellular cyclic AMP evoked by C5adesArg and C3a; it is prevented by GDP beta S (5 X 10(-4) M) which blocks activation of adenylate cyclase, and prolonged by agents which sustain cyclic AMP elevations, namely 5 X 10(-4) M theophylline and 5 X 10(-4) M) GTP gamma S. PMID- 2983807 TI - Pre- and post-junctional actions of prostaglandin I2, carbocyclic thromboxane A2 and leukotriene C4 in dog tracheal tissue. AB - Effects of carbocyclic thromboxane A2 (cTxA2), prostacyclin (PGI2) or leukotriene C4 (LTC4) on the membrane and contractile properties of the smooth muscle cells and on the excitatory neuro-effector transmission in the dog trachea were observed by means of the microelectrode, double sucrose gap and tension recording methods. cTxA2, PGI2 or LTC4 at a concentration of 10(-7)M had no effect on the membrane potential of smooth muscle cells of the dog trachea. At 10(-6)M, cTxA2 and LTC4 slightly depolarized, and PGI2 hyperpolarized the membrane. cTxA2 (greater than 2.7 X 10(-10)M) evoked a sustained contraction, while the amplitude of the twitch contractions evoked by field stimulation in the presence of indomethacin (10(-6)M) and propranolol (10(-6)M) was inhibited, dose-dependently. PGI2 (greater than or equal to 2.7 X 10(-7)M) reduced the muscle tone and the amplitude of twitch contractions evoked by field stimulations. cTxA2 or PGI2 (10( 10)-10(-7)M) reduced the amplitude of the excitatory junction potentials (e.j.ps) evoked by field stimulation with no change in the membrane potential, input membrane resistance or the sensitivity of the muscle cells to acetylcholine (ACh). LTC4 (1.6 X 10(-8)M) evoked a sustained contraction of the dog trachea; however, this agent did not affect either the amplitude of the twitch contractions or the e.j.ps evoked by field stimulation. The amplitude of the e.j.p. was dependent on the external concentration of Ca2+, and the inhibitory actions of cTxA2 on e.j.ps were partly overcome by increasing the concentrations of [Ca]o. When the amplitudes of e.j.ps were plotted against [Ca]o on a double log scale, the above relation yielded a straight line with a slope of 1.7 or 1.0, in the absence or presence of cTxA2, respectively. After treatment with Ca2+-free 2 mM EGTA-containing solution, cTxA2 or LTC4 did not evoke a contraction in the dog trachea, whereas ACh (10(-7)-10(-6)M) did. These results indicate that cTxA2 and PGI2 have dual actions on pre- and post-junctional membranes of the dog tracheal tissue, i.e. both agents inhibit the excitatory neuro-effector transmission in the dog trachea, presumably by inhibiting the release of ACh from the vagal nerve terminal. cTxA2 and LTC4 or PGI2 evoke contraction or relaxation of the muscle tissue, respectively, apparently through direct actions on the smooth muscle cells. PMID- 2983808 TI - Ion dependence of the release of noradrenaline by tetraethylammonium and 4 aminopyridine from cat splenic slices. AB - Cat splenic slices prelabelled with [3H]-noradrenaline were incubated in oxygenated Krebs-bicarbonate solution at 37 degrees C, and the spontaneous total 3H release into different incubation media monitored. In normal Krebs bicarbonate solution, the spontaneous tritium fractional release amounted to 3.7% of the tissue radioactivity content per 5 min collection period. Tetraethylammonium (TEA) increased spontaneous transmitter release in a concentration-dependent manner; the release was maximal at 30 mM and was 3.5 times the basal release. 4 Aminopyridine (4-AP) also enhanced the spontaneous release of tritium. The response increased linearly with 4-AP concentration (1-10 mM). With 10 mM 4-AP, the release was as much as 6 times the basal transmitter release. Guanidine was much less potent than either TEA or 4-AP. The secretory response to TEA or 4-AP was little affected by changes in external Ca2+, Mg2+, Na+, Cl-, H2PO4- or by tetrodotoxin. However, transmitter release evoked by TEA or 4-AP strongly depended upon the concentration of HCO3- of the incubation solution; in fact, the secretory response varied almost linearly between 1 and 25 mM HCO3-. The mechanisms underlying these effects are probably related to the well-known ability of TEA and 4-AP to block K+ conductance that would cause depolarization of the splenic sympathetic nerve terminals. The HCO3- requirements for the secretory response are probably related to the ability of CO2/HCO3- solutions to mobilize and release Ca2+ from intracellular organelles. PMID- 2983809 TI - Some anticonvulsant drugs alter monoamine-mediated behaviour in mice in ways similar to electroconvulsive shock; implications for antidepressant therapy. AB - The effects in mice of administration of the anticonvulsants, progabide, sodium valproate, diazepam, carbamazepine and phenytoin on 5-hydroxytryptophan (5-HTP) induced head-twitch, apomorphine-induced locomotion, clonidine-induced sedation, and beta-adrenoceptor and 5-HT2 receptor number have been examined. Repeated progabide administration (400 mg kg-1, i.p. twice daily for 14 days) enhanced the head-twitch response the effect lasting for over 8 days after the last dose, and also increased 5-HT2 receptor number in frontal cortex. Progabide (400 mg kg-1, i.p.) enhanced the head-twitch response when given once daily for 10 days and when given intermittently (5 times over 10 days) but not after 1 day of administration. Repeated Na valproate (400 mg kg-1, i.p.) also increased the 5 HTP-induced head-twitch response and 5-HT2 receptor number in the frontal cortex when given twice daily for 14 days, but no behavioural enhancement was seen after 10 days' treatment. Diazepam (1.25 mg kg-1, i.p.) twice daily for 14 days increased the head-twitch response and 5-HT2 receptor number. Repeated progabide and valproate (but not diazepam) administration attenuated the sedation response to the alpha 2-adrenoceptor agonist, clonidine (0.15 mg kg-1) but neither drug altered beta-adrenoceptor number in the cerebral cortex. No changes in apomorphine-induced locomotor behaviour were seen after progabide, valproate or diazepam. Repeated carbamazepine (20 mg kg-1) or phenytoin (40 mg kg-1) administration failed to alter any of the biochemical or behavioural parameters listed above. Like repeated electroconvulsive shock (ECS), progabide altered the head-twitch response, clonidine-induced sedation response and 5-HT2 receptor number. Unlike repeated ECS, it did not alter beta-adrenoceptor number or the apomorphine-induced locomotor response. These data suggest that ECS may produce some changes in monoamine function by altering GABA metabolism as has previously been postulated. PMID- 2983810 TI - Comparison of the action of cholinomimetics and pentagastrin on gastric secretion in dogs. AB - Stimulation of acid secretion by muscarinic cholinomimetic agents depended on the periodic interdigestive activity of the stomach. This explains the peak and following fade. Pentagastrin stimulated gastric secretion after a fixed interval and did not depend on the interdigestive activity. Neither gastrin nor methacholine directly contracted the gallbladder in the doses used. A sustained secretion of gastric acid and pepsin, such as follows a meal, required both hormonal stimulation and gastric distension. The magnitude of the acid response from Heidenhain pouches following meals suggested that pentagastrin doses commonly used experimentally greatly exceed the physiological. PMID- 2983813 TI - Studies on the stereoselectivity of the P2-purinoceptor on the guinea-pig vas deferens. AB - ATP, 2-chloro-ATP, 2-methylthio-ATP and their unnatural L-enantiomers, Rp and Sp diastereoisomers of the ATP phosphorothioate analogues, ATP alpha S and ATP beta S, were tested on the guinea-pig vas deferens. The 2-substituted analogues of ATP were no more effective than ATP in causing contraction of the vas deferens. However, stereoselectivity was observed with each pair of enantiomers of ATP, 2 chloro-ATP and 2-methylthio-ATP. No stereoselectivity was observed for the phosphorothioate analogues. Rp- and Sp-ATP beta S were more effective than ATP at eliciting contractions of the vas deferens. These results show that unlike the P2 purinoceptor mediating excitatory responses in the guinea-pig bladder, the P2 purinoceptor mediating contraction in the guinea-pig vas deferens displays stereoselectivity. PMID- 2983812 TI - Hypothermia-induced supersensitivity to adenosine for responses mediated via A1 receptors but not A2-receptors. AB - Four isolated tissues were examined in which the responses to adenosine are mediated via either A1- or A2-receptors. The responses examined were the inhibition of cholinergic transmission of field-stimulated guinea-pig ileum (A1), inhibition of noradrenergic transmission of field-stimulated rat vas deferens (A1), inhibition of developed tension of rat paced left atria (A1) and relaxation of carbachol-contracted guinea-pig trachea (A2). Cumulative concentration response curves for adenosine and 2-chloroadenosine were constructed at 37, 30 or 27 degrees C. When plotted as a percentage of the maximum response, the concentration-response curves were displaced to the left by cooling in the ileum, vas deferens and atria, indicative of supersensitivity. This increase in sensitivity does not arise from inhibition of uptake or deamination by cooling, since it occurs equally for adenosine and 2-chloroadenosine, the latter being immune to these processes. In contrast, the sensitivity of the trachea was not affected (2-chloroadenosine) or reduced (adenosine) by cooling. Thus responses mediated via adenosine receptors of the A1 subtype exhibit hypothermia-induced supersensitivity, whereas those mediated via A2-receptors do not. This suggests a fundamental temperature-dependent difference between the two adenosine receptor subtypes. PMID- 2983811 TI - Nitroglycerine- and isoprenaline-induced vasodilatation: assessment from the actions of cyclic nucleotides. AB - To investigate the vasodilator actions of nitroglycerine and isoprenaline, the effects of these agents, dibutyryl cyclic AMP (db cyclic AMP) and 8-bromo cyclic GMP (8-Br cyclic GMP) on intact muscle tissue, and cyclic AMP and cyclic GMP on skinned muscle of the rabbit mesenteric artery were investigated. In porcine coronary artery, nitroglycerine (greater than 0.1 microM) increased the production of cyclic GMP with no change in the amount of cyclic AMP, while isoprenaline (greater than 0.1 microM) significantly increased the production of cyclic AMP with no change in the amount of cyclic GMP. In the rabbit mesenteric artery, nitroglycerine or isoprenaline inhibited the tonic component of the 39 mM [K]o-induced contraction to a greater extent than the phasic component. Nitroglycerine and 8-Br cyclic GMP showed a stronger inhibitory action on the K induced contraction than did isoprenaline and db cyclic AMP. The sources of Ca utilized for the generation of contraction by noradrenaline and caffeine were estimated to be the same as those determined from the amplitudes of contractions evoked in Ca-free solution by various concentrations of noradrenaline or caffeine. In intact muscle tissues, the effects of nitroglycerine or 8-Br cyclic GMP on the amount of Ca stored in cells were estimated from the caffeine-induced contraction in Ca-free solution. Both agents inhibited the contractions due to a reduction in the amount of Ca in the cells. When the effects of isoprenaline or db cyclic AMP were observed, both agents inhibited the caffeine-induced contraction but the accumulation of Ca into cells was greater than the control. In saponin skinned muscles, the pCa-tension relationship in the presence of cyclic AMP and cyclic AMP-dependent protein kinase (cyclic AMP-PK) shifted to the right and to a lower level in comparison with the control. Applications of cyclic GMP with cyclic GMP-dependent protein kinase (cyclic GMP PK) also inhibited the contraction induced by low concentrations of Ca. In skinned muscles, cyclic AMP exhibited dual actions on Ca store sites, i.e. in the presence of high concentrations of Ca or prolonged superfusion of Ca, cyclic AMP reduced the amount of Ca due to activation of the Ca-induced Ca release mechanism by excess accumulation of Ca. On the other hand, cyclic GMP consistently inhibited the amplitude of the caffeine-induced contraction due to a reduction in the amount of Ca in the store sites. 8 These results indicate that nitroglycerine and isoprenaline increase the amount of cyclic GMP and cyclic AMP, respectively. The main effect of cyclic GMP is activation of Ca extrusion, thus reducing the amount of Ca stored in the cell, while the main effect of cyclic AMP is to increase the amount of Ca stored in the cell. Both cyclic AMP with cyclic AMP-PK and cyclic GMP with cyclic GMP-PK inhibit the phosphorylation of myosin. Consequently both cyclic nucleotides reduce the free Ca in the myoplasm and promote relaxation, but by different mechanisms. PMID- 2983815 TI - Evidence for neuro-effector transmission through postjunctional alpha 2 adrenoceptors in human saphenous vein. AB - The effects of the alpha 1-adrenoceptor antagonist prazosin and the alpha 2 adrenoceptor antagonist yohimbine were examined against stimulation-evoked contractions in human isolated saphenous veins. The concentration of yohimbine producing 30% inhibition of stimulation-evoked contractions (IC30) was 13.2 nM, whereas the IC30 of prazosin was greater than 250 nM. The inhibition of stimulation-evoked contractions by yohimbine was not prejunctionally mediated since yohimbine (0.01-0.1 microM) significantly potentiated the stimulation evoked overflow of tritium in tissues pre-incubated with [3H]-noradrenaline. The high potency of yohimbine and the low potency of prazosin indicate that neuro effector transmission in human saphenous vein is mediated predominantly by postjunctional alpha 2-adrenoceptors. PMID- 2983814 TI - Effect of adenosine triphosphate on the sensitivity of the nicotinic acetylcholine-receptor in the bullfrog sympathetic ganglion cell. AB - The effects of adenosine triphosphate (ATP) and related compounds on the sensitivity of the nicotinic acetylcholine (ACh)-receptor of bullfrog sympathetic ganglion cells were analysed electro-physiologically. ATP in concentrations between 0.05 and 2 mM increased the amplitudes of the potentials and currents induced by ACh, and carbachol-induced currents. Compared with ATP, ADP was less potent in producing augmentation of the carbachol-induced current by one order of magnitude. AMP, cyclic AMP and adenosine had no appreciable effect. Analysis of this ATP effect, based on Michaelis-Menten type kinetics, revealed that ATP increased the maximum response (Vmax) of the dose-response curve of ACh currents without an appreciable effect on the affinity (Km) of ACh for its receptor. It is suggested that ATP increased the receptor sensitivity by acting on an allosteric site of the nicotinic ACh receptor-ionic channel complex which, thus, may be linked to an ATP receptor, probably of the P2-receptor type (Burnstock, 1981). PMID- 2983816 TI - Nuclear magnetic resonance (NMR): I. Imaging biochemical change. AB - Nuclear magnetic resonance (NMR) imaging of hydrogen in body water and fat is proving valuable in clinical investigation of the brain. An introduction to the technique and to the biological significance of the images is presented here. The 'multi-parameter' nature of these images is stressed, and the individual parameters described. NMR imaging may well be of value in investigating the pathology of organic and functional psychoses. PMID- 2983817 TI - Nuclear magnetic resonance (NMR). II. Imaging in dementia. AB - Proton NMR imaging of the brain is rapidly becoming established as a useful investigative tool in medicine. This paper examines the usefulness of the NMR parameters--spin-lattice relaxation time (T1) and proton density (PD)--in differentiating groups of patients with senile dementia of Alzheimer type (SDAT) and multi-infarct dementia (MID) from each other, and from elderly controls. T1 values increase with severity of dementia. NMR parameters may also be of use in localising regions of brain damage. PMID- 2983818 TI - Epidermal growth factor receptors on human breast cancers. AB - Epidermal growth factor (EGF) is a potent mitogen for many tissues including breast. The receptor for EGF (EGF-r) has the same peptide sequence as a known oncogene. Sixty-one human breast cancers and nine associated lymph node metastases have been examined for the presence of EGF receptors by competitive binding and immunocytochemical techniques. Oestrogen receptor analysis on the same specimens was performed by the dextran coated charcoal method and compared with EGF-r status. These were correlated with the clinical findings and histological data. An inverse relationship between EGF-r and ER was found (chi 2= 7.81, P less than 0.01) with a higher incidence of EGF-r positive tumours in the metastatic group (chi 2 = 14.51, P less than 0.001). EGF-r positive tumours were of poor differentiation and had characteristics associated with poor prognosis. PMID- 2983819 TI - The Klippel-Trenaunay syndrome: clinical, radiological and haemodynamic features and management. AB - Forty-nine patients with the Klippel-Trenaunay syndrome have been studied. Sixty eight per cent have a superficial, embryological venous channel on the lateral aspect of the limb, 25 per cent have had one or more severe spontaneous haemorrhages from dilated varices and 22 per cent have suffered a venous thrombo embolism. Twenty-nine per cent have had episodes of rectal bleeding or haematuria associated with pelvic angiomas. Foot volumetry (n = 31) showed that calf emptying was decreased in only five patients and only one of these had phlebographic evidence of deep venous obstruction, which conflicts with previous findings. The rate of calf refilling was increased in 81 per cent and phlebography demonstrated incompetent communicating veins in 45 per cent of patients. No patient had clinical evidence of an arteriovenous fistula, and arteriography (n = 22) and calf blood flow (n = 33) were normal, but lymphangiography (n = 14) showed lymphatic hypoplasia in 55 per cent of limbs. Eighty-eight operations have been performed on 38 patients but symptoms persist in 90 per cent. We suggest that surgery should be limited to the excision of localized symptomatic abnormalities and that the best form of control is provided by graduated compression stockings. PMID- 2983821 TI - Peripheral neuropathy during treatment with almitrine. PMID- 2983820 TI - Seroepidemiology of HTLV-III antibodies in a remote population of eastern Zaire. AB - A human retrovirus--human T cell lymphotropic virus-III (HTLV-III)--has recently emerged as the probable cause of acquired immunodeficiency syndrome (AIDS). In May 1984, 250 outpatients at a hospital in a remote area of eastern Zaire were surveyed for AIDS type illnesses and the prevalence of antibodies against HTLV III determined by an enzyme linked immunosorbent assay using disrupted whole HTLV III virus as the antigen. No clinical cases of AIDS were diagnosed among these patients. Overall, 31 (12.4%) had clearly positive ratios (greater than or equal to 5.0) and a further 30 (12.0%) had borderline ratios (3.0- less than 5.0). Western blots of serum samples from subjects with antibodies yielded bands consistent with HTLV-III as found in American patients with AIDS and members of groups at risk of AIDS. The prevalence of antibody was highest in childhood (p = 0.02); among adults prevalence rose slightly with age. HTLV-III antibodies were more common among the uneducated (p = 0.006), agricultural workers (p = 0.03), and rural residents (p = 0.006), but the Western blot bands were generally weak in this group. By contrast, one urban resident had strong bands. The relatively high prevalence of antibodies among the rural poor in this area of Zaire suggests that HTLV-III or a closely related, cross reactive virus may be endemic in the region. A different natural history of infection, perhaps in childhood, may also explain the findings. PMID- 2983822 TI - Air in the bones: multifocal anaerobic osteomyelitis associated with oat cell carcinoma. PMID- 2983823 TI - Pattern and extent of demyelination in the optic nerves of mice infected with Semliki Forest virus and the possibility of axonal sprouting. AB - The optic nerves of all mice infected with Semliki Forest virus (SFV) showed patchy demyelination, although the extent of demyelination varied between animals. There was a significant increase in small diameter unmyelinated fibres in the optic nerves of SFV infected mice, paralleled by a loss of small myelinated fibres, suggesting that the smaller myelinated fibres may be more susceptible to virus-induced demyelination. The small unmyelinated fibres were more numerous in the peripheral region of the optic nerve trunk, where blood vessels are found in greater numbers. In addition, a particularly large increase in the proportion of unmyelinated fibres which had diameters of less than 0.2 microns suggested the possibility of axonal sprouting. PMID- 2983824 TI - Time course of miniature postsynaptic potentials at the Mauthner fiber giant synapse of the hatchetfish. AB - The hatchetfish Mauthner fiber is presynaptic to 8-14 large myelinated axons in the medulla; the large ('giant') synapses formed by these fibers appear to be nicotinic cholinergic. Miniature postsynaptic potentials (mPSPs) were recorded from single identified synapses. The mPSPs were averaged to more accurately determine their shape; the rise time was approximately 70 microseconds, and the fall usually was biphasic with time constants of decay for the two phases of 280 and 800 microseconds. In 25% of the records analyzed a third, slow tail of decay was seen which had an average decay constant of 4.2 ms. The biphasic decay of mPSPs largely accounts for the similar shape of the postsynaptic current following a presynaptic impulse, which is described in the accompanying paper. PMID- 2983825 TI - Postsynaptic currents at the Mauthner fiber giant synapse of the hatchetfish. AB - Postsynaptic currents (PSCs) at the giant synapse between Mauthner and giant fibers of the hatchetfish Gasteropelecus were studied under voltage clamp. This axo-axonic synapse lies in the central nervous system beneath the floor of the 4th ventricle where electrodes can be closely positioned both pre- and postsynaptically. Transmission is nicotonic cholinergic. The PSCs produced by Mauthner fiber impulses rise rapidly to a peak and decay in two phases; an early more rapid phase is followed by a late slower phase. The slope conductance of the peak amplitude of the PSCs declines at more inside positive potentials. The late phase of decay is exponential and voltage dependent, becoming faster for PSCs evoked at more inside positive potentials. At potentials positive to about -40 mV the late phase merges with the early phase. The decay rate constant of the slowest phase is exponentially related to voltage for potentials negative to about -10 mV, but becomes less voltage dependent for more positive potentials. The peak current is independent of whether it is evoked during inward or outward active currents of the electrically excitable membrane, and two phase decays are observed in PSCs of reduced quantal content. Thus, changes in slope conductance and two phase decays are not due to series resistance or interactions between quanta. PSCs can be modeled by a 3 state reaction scheme in which closed channels open when they bind transmitter and then can pass to a second closed state with receptor still bound such that they must return through the open state before losing their transmitter and returning to the resting, closed state. PMID- 2983826 TI - The effects of a glycine antagonist (strychnine) on cortically induced rhythmical jaw movements in the anesthetized guinea pig. AB - We have investigated the effects of systemic administration of a glycine antagonist, strychnine, on rhythmical jaw movements (RJMs) induced by repetitive stimulation of the masticatory area of the guinea pig cerebral cortex. It was found that after strychnine administration (0.4 mg/kg), the frequency of the cortically induced RJMs was minimally affected, whereas the burst durations of the digastric EMG during the jaw opening phase of the RJM cycle were dramatically increased. These data suggest that the neuronal network (central pattern generator (CPG)) which is responsible for the production of rhythmical jaw movements is not critically dependent upon glycine for the genesis of the basic oscillatory rhythm. On the other hand, glycine synapses are involved with the neuronal mechanisms which are responsible for controlling the burst durations of the digastric muscle during the jaw opening phase of each rhythmic jaw movement cycle. PMID- 2983827 TI - Localization of 2',3'-cyclic nucleotide 3'-phosphodiesterase on cultured Schwann cells. AB - Primary cultures of Schwann cells were labeled by indirect immunofluorescence using an antibody directed against 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase). Schwann cells which had been maintained in culture for 8 weeks were labeled with this antibody. Immunoblot analysis of Schwann cell homogenates revealed a single band with a molecular weight of 54,000 daltons which corresponded to a single immunoreactive polypeptide present in myelin prepared from rat sciatic nerve. The subcellular localization of CNPase was examined by fractionation of cultured Schwann cell homogenates with linear sucrose gradients. The distribution of CNPase paralleled that of 5'-nucleotidase, a putative marker for plasma membranes. These results suggest that CNPase is localized on the plasma membranes of Schwann cells and is expressed by the cells in the absence of an axonal stimulus. PMID- 2983828 TI - Neural activity originating from a neuroma in the baboon. AB - Single nerve fiber recordings were obtained from traumatically induced neuromas of the superficial radial nerve in baboons 1-7 months after injury. Eight to 18% of the fibers had spontaneous activity, and 67% of these were unmyelinated. Myelinated as well as unmyelinated fibers responded to mechanical stimulation of the neuroma whereas no fibers responded to similar stimulation of the normal nerve. Apparent crosstalk of action potential activity between fibers at the neuroma was observed which could be due to electrical coupling, though retrograde sprouting is another possible explanation. These abnormalities in neural activity originating from a neuroma in the primate are qualitatively similar to those noted in other species and may provide an explanation for certain abnormal sensory phenomena associated with peripheral nerve injury. PMID- 2983829 TI - alpha-Melanotropin, beta-endorphin and adrenocorticotropin-like immunoreactivities are colocalized within duodenal myenteric plexus perikarya. AB - The opioid peptide beta-endorphin co-exists with alpha-melanotropin and adrenocorticotropin within myenteric neuronal cell bodies of the rat duodenum. Adjacent serial sections through the myenteric and submucous plexus have been stained alternately with antisera directed against alpha-melanotropin, beta endorphin and adrenocorticotropin. This resulted in nearly superimposable immunofluorescences for the 3 peptides within neuronal cell bodies of the myenteric plexus. An alpha-melanotropin staining was always linked by an immunofluorescence for beta-endorphin, adrenocorticotropin and vice versa. These staining patterns were not seen in the submucous plexus. Some tangentially cut nerve fibers running through the longitudinal muscle layer revealed a coexistence of adrenocorticotropin and alpha-melanotropin. The colocalization of beta endorphin, alpha-melanotropin and adrenocorticotropin within the same perikarya may reflect a physiological role for the 3 peptides in the nervous system of the rat duodenum. PMID- 2983830 TI - Synaptic amplification by active membrane in dendritic spines. AB - The suspected functional role of dendritic spines as loci of neuronal plasticity (possibly memory and learning) is greatly enriched when active membrane properties are assumed at the spine head. Computations with reasonable electrical and structural parameter values (corresponding to an optimal range for spine stem resistance) show that an active spine head membrane can provide very significant synaptic amplification and also strongly non-linear properties that could modulate the integration of input from many afferent sources. PMID- 2983831 TI - Development of the rat pineal alpha 1-adrenoceptor. AB - Pineal alpha 1-adrenoceptors in rats from 19 days of gestation until 11 months of age were studied using [125I]iodo-2-[beta-(4 hydroxyphenyl)ethylaminomethyl]tetralone ( [125I]HEAT). The number of specific [125I]HEAT binding sites increased markedly between 18 days of gestation (101.7 +/- 13.1 fmol/mg protein) and 10 days of age (336.2 +/- 34.3 fmol/mg protein). A significant decline occurred after 1 month of age. A saturation study showed similar changes in receptor density with age (Bmax; 20 days of gestation, 130.5 fmol/mg protein; 35 days old, 288.1 fmol/mg protein) but no difference in Kd (58.4 pM at both -1 and +35 days). The developmental appearance of the pineal alpha 1-adrenoceptor and the decline in its density with age are remarkably similar to changes reported for pineal beta-adrenoceptors. PMID- 2983832 TI - Activity dependent plasticity of postsynaptic density structure in the ventral cochlear nucleus of the rat. AB - Young adult male rats were anaesthetised with urethane and exposed to either 24 h of silence or 24 h of repetitive 77 dB tones in a sound-proofed anechoic chamber. The influence of these two conditions on the ultrastructure of the synaptic appositions made by auditory afferents in the anterior ventral cochlear nucleus (end bulbs of Held) was compared. Both the cross-sectional area and the mean thickness of the postsynaptic density (PSD) in the rats exposed to tones were significantly reduced when compared with rats maintained in silence. Similarly, the degree of curvature of the apposition was significantly reduced. The results of these experiments provide further evidence that the postsynaptic density material is a plastic structure significantly influenced by the amount of activity in the presynaptic element. PMID- 2983833 TI - Synaptic responses of neurons of the nucleus tractus solitarius in vitro. AB - Postsynaptic responses of neurons in the nucleus tractus solitarius (NTS) have been studied in an in vitro slice preparation using extra- and intracellular recording. Single or paired pulse stimulations were delivered to afferent fibers within the tractus solitarius (TS) to activate orthodromic responses in these neurons. Most NTS neurons displayed an initial synaptic excitation followed by inhibition of spontaneous or evoked firing lasting up to 150-200 ms after stimulation. Excitatory postsynaptic potentials (EPSPs), recorded intracellularly, were increased in amplitude by membrane hyperpolarization. Large afterhyperpolarizations followed action potentials triggered by the EPSPs or evoked by intracellular current injections. Intracellular evidence for synaptic inhibition within the NTS included: (1) the presence, after Cl-injection, of flurries of spontaneous PSPs likely to be inverted inhibitory postsynaptic potentials; (2) reduction of the size of a test EPSP by a previous subthreshold TS conditioning volley; and (3) hyperpolarizing PSPs recorded in some neurons. Other NTS neurons exhibited prolonged excitatory responses to TS stimulation and could be local inhibitory interneurons. These results may help specify synaptic mechanisms in the NTS that could play an integrative role in the relay of visceral sensory inputs to higher order effectors. PMID- 2983834 TI - Long-term potentiation in the dentate gyrus: effects of noradrenaline depletion in the awake rat. AB - The chronic rat preparation was utilized to study the effects of noradrenaline (NA) depletion on field potentials recorded from the hilus of the fascia dentata. Both single pulses and high-frequency trains were applied to the perforant path (PP). The effects of NA depletion on baseline responses as well as on long-term potentiation (LTP) were examined. Reduced NA levels resulted in an increase in the population spike amplitude and a depression of the population excitatory postsynaptic potential (EPSP). Depleted animals showed significantly higher levels of LTP of the population EPSP, but reduced levels of population spike LTP (measured 13-15 min after tetanization). There were, however, no differences in LTP levels 1 week after the potentiation tests. These results demonstrate that NA levels do not affect that component of LTP which can persist for several weeks. PMID- 2983835 TI - Noradrenergic inhibitors cause accumulation of nuclear progestin receptors in guinea pig hypothalamus. AB - A series of experiments was performed to study the possible behavioral relevance of the apparent regulation of hypothalamic cytosol progestin receptors by noradrenergic transmission in female guinea pigs. In the first experiment, ovariectomized guinea pigs were injected with estradiol benzoate followed 36 h later by the dopamine-beta-hydroxylase inhibitor, U-14,624 or vehicle. Twelve h later, they were injected with progesterone and tested hourly for sexual behavior. Six h after the progesterone injection, a time at which inhibition of sexual behavior by the U-14,624 was confirmed, they were killed, and cytosol and nuclear progestin receptors were assayed in the hypothalamus and cerebral cortex. No difference was seen in the concentration of progestin receptors after drug treatment in these animals that also received a progesterone injection. In subsequent experiments, it was found that the U-14,624-inhibition of progesterone facilitated sexual behavior is not accompanied by an inhibition of nuclear progestin receptor accumulation. Furthermore, it was found that the decreased cytosol progestin receptor level caused by U-14,624 prior to progesterone injection was accompanied by an increase in the concentration of nuclear progestin receptors. The increase in nuclear progestin receptors was also seen after treatment with the alpha-adrenergic antagonist, prazosin, U-14,624 does not compete with [3H]R 5020 for binding to the progestin receptor, suggesting that it does not directly cause translocation of progestin receptors. The results of these experiments suggest that the decrease in the concentration of cytosol progestin receptors caused by noradrenergic inhibitors is not due entirely to an interference with the formation of cytosol progestin receptors. Rather, it seems that these drugs, in some way, also cause the accumulation of progestin receptors in cell nuclei. Furthermore, they suggest that the mechanism by which these drugs inhibit sexual behavior may not be by interference with the progestin receptor system. PMID- 2983836 TI - Correlation of rates of calcium entry and endogenous dopamine release in mouse striatal synaptosomes. AB - The time course of simultaneous Ca2+ entry and endogenous dopamine release was examined in mouse striatal synaptosomes depolarized by 30 mM KCl. Ca2+ entry and endogenous dopamine release exhibited fast and slow phase processes. The fastest rates occurred between 0 and 1 s. Ca2+ uptake and dopamine release dropped off quickly with 5-15 s rates at 13 and 10%, respectively, of the 0-1 s rate. Both processes were maintained at relatively high rates at the 1-3 and 3-5 s intervals suggesting mixed fast and slow phase processes. Uptake of Ca2+ and release of dopamine occurred in parallel over the entire 30 s measurement period; however, approximately 70% of the Ca2+ uptake and dopamine release occurred within the first 5 s following depolarization. A calculated ratio of Ca2+ entry versus dopamine release showed that approximately 1-2 Ca2+ ions were required to cause the release of one dopamine molecule. This ratio remained constant from 1 to 15 s following depolarization. Our results suggest that Ca2+ entry is coupled to endogenous dopamine release for both the fast and slow phase process. PMID- 2983837 TI - The glycine receptor in the mutant mouse spastic (spa): strychnine binding characteristics and pharmacology. AB - There is a marked deficit in the binding of the glycine receptor antagonist strychnine to the CNS of the mutant mouse spastic. The characteristics and pharmacology of [3H]strychnine binding to washed homogenates of spastic and littermate control spinal cord and brainstem were investigated to determine the nature of this defect. The maximal binding of [3H]strychnine to spastic homogenates is approximately 20% the value obtained from littermate control homogenates; the affinity of [3H]strychnine binding is approximately 25% lower than littermate control values. The pharmacology of [3H]strychnine binding has the same rank order of potency in spastic and littermate control mice; however, there are small differences in the potency of several compounds in spastic vs littermate control animals. These results indicate that the glycine receptor alteration seen in the spastic mutant mouse is primarily due to a decrease in receptor number. They also suggest that the pharmacological characteristics of the glycine receptor in spastic are different from littermate control. The data do not permit a distinction between whether the strychnine binding sites found in spastic represent the same population seen in littermate control animals or are a subpopulation that is spared by the mutation. PMID- 2983838 TI - Benzodiazepine receptors in the visual structures of monocularly deprived rats. Effect of light and dark adaptation. AB - In 25-day-old rats with one eyelid sutured at the age of 10 days, the binding of [3H]flunitrazepam in the visual structures (retina, lateral geniculate nucleus, superior colliculus, visual cortex) and frontal cortex was determined. Monocular visual deprivation (MD) resulted in a significant decrease of the [3H]flunitrazepam binding in the retina of the open eye to about 76% of the control value. No changes in [3H]flunitrazepam binding were detectable under these conditions in the central visual structures examined and the non-visual cortical region. Scatchard analysis indicated that the changes found in the retina of the open eye of MD rats are due to a decreased binding affinity only, the maximum receptor number being unaffected. Eight hours after re-opening the sutured eyelid of 25-day-old MD rats, benzodiazepine binding in the open eye was increased to the control level, whereas the binding in the retina of the re opened eye remained unchanged in comparison to control animals. Dark adaptation of 25-day-old control rats resulted in an increased [3H]flunitrazepam binding in the retina by 28% compared to that detectable in the retina of light-adapted animals. In contrast, dark-adaptation of MD rats did not affect [3H]flunitrazepam binding in the retina of both eyes in comparison to that found in the corresponding retina of light-adapted MD animals. The data obtained suggest a physiological coupling between both retinas, possibly mediated through centres inside of the central nervous system. PMID- 2983839 TI - Topographical distribution of pro-opiomelanocortin-derived peptides (ACTH/beta END/alpha-MSH) in the rat median eminence. AB - The detailed distribution of adrenocorticotropin (ACTH), beta-endorphin (beta END) and alpha-melanotropin (alpha-MSH) immunoreactivity was examined in the rat median eminence (ME) and pituitary stalk using light microscopic immunocytochemistry and radioimmunoassay (RIA). Nerve fibers and varicosities immunoreactive for ACTH/beta-END/alpha-MSH had identical distributions in the ME suggesting that they are part of the same arcuate proopiomelanocortin neuronal (POMC) system. The quantitative image analysis of POMC immunoreactive varicosities in the ME indicates no significant differences between the various rostro-caudal segments. In the main (preinfundibular) portion of the ME, a moderate density of immunoreactive elements was located in the lateral part of the internal zone and throughout the postinfundibular ME. Very few scattered varicosities were observed in the neurohemal (external) zone and in the pituitary stalk. By RIA, alpha-MSH is present in a substantially higher concentration than ACTH and beta-END throughout the ME. Knife cuts between the arcuate nucleus and ME indicate that proopiomelanocortin (POMC) fibers enter the ME in its whole rostro-caudal extent. Thus POMC neurons seem to provide innervation of structures in the internal zone but not in the neurohemal/external/zone where the portal capillary system is located. Moreover, the observation that the density of immunoreactive elements is substantially lower in the pituitary stalk than in the ME, suggests that the majority of immunoreactive fibers in the internal zone are not fibers of passage directed towards the neurohypophysis. PMID- 2983840 TI - beta-Adrenoceptor stimulation-induced increase in the number of alpha 2 adrenoceptors of cerebral cortical membranes in rats. AB - Effects of pre-treatment of synaptic membranes with beta-adrenoceptor agonists and cholera toxin on [3H]clonidine and [3H]yohimbine binding were examined in rat cerebral cortex. Pre-incubation of cerebral cortical membranes with isoproterenol (10 or 200 microM) or dobutamine (1, 10 or 200 microM) at 37 degrees C for 40 min caused a significant elevation of specific [3H]clonidine binding but treatment with salbutamol (10 or 200 microM) did not. Scatchard analysis showed that 200 microM isoproterenol treatment resulted in a significant elevation of high affinity component of [3H]clonidine binding which was significantly decreased by the addition of 10 microM GTP. A significant elevation in high affinity [3H]clonidine binding was observed by treatment with 100 micrograms/ml cholera toxin, while a significant decrease in low affinity one was by the treatment. Specific [3H]yohimbine binding was also elevated by 10 or 200 microM isoproterenol treatment. It is suggested that stimulation of beta-receptors, presumably beta 1-subtype, could elevate the number of agonist and antagonist binding sites in alpha 2-receptors in synaptic membranes by partially mediated by stimulatory and/or inhibitory GTP binding regulatory proteins. PMID- 2983841 TI - Extracranial vasodilation mediated by vasoactive intestinal polypeptide (VIP). AB - Pooled antisera to vasoactive intestinal polypeptide were used to block neurogenic extracranial vasodilatation elicited from either brainstem (locus coeruleus) or pterygopalatine ganglion stimulation in the cat. Vasodilatation was not inhibited by sham immune sera, or by antisera to bradykinin or substance P. The efferent pathway for vasodilatation from the locus coeruleus traverses the facial nerve (greater superficial petrosal branch) and the pterygopalatine and otic ganglia. Its blockade demonstrates a novel action of a peptide transmitter in the expression of a central neurogenic response. PMID- 2983843 TI - Antagonist activity of methyl-substituted analogues of 2-amino-4 phosphonobutanoic acid in the hippocampal slice. AB - Four monomethyl-substituted analogues of 2-amino-4-phosphonobutanoic acid (APB), an antagonist of excitatory pathways in the central nervous system, were prepared in order to investigate the steric requirements of the APB receptor. Methyl groups were incorporated at the amino, alpha-, beta-, and gamma-positions. The beta- and gamma-methyl-substituted analogues of APB were found to be moderately potent antagonists in excitatory synapses of the hippocampal perforant path, as judged by extracellular recording techniques, while the N- and alpha-methyl substituted analogues had much lower potencies. All of these APB analogues had very low potencies in the Schaffer collateral pathway. The APB receptors in the perforant path displayed more tolerance of methyl-substitution at the beta- and gamma-positions of APB than at the amino or alpha-positions in this system. PMID- 2983842 TI - Cations differentially affect subpopulations of L-glutamate receptors in rat synaptic plasma membranes. AB - Several cations were examined for their ability to specifically affect one of the 3 L-glutamate (L-Glu) binding sites in rat forebrain synaptic plasma membranes (i.e. Na+-dependent, Cl--dependent and Cl--independent). Na+-dependent binding was potently inhibited by K+ and NH4+ ions. Other monovalent cations tested (Cs+, Li+, triethylammonium) had no effect on this binding site. Polyvalent cations (Co2+, Ni2+, Cu2+, Zn2+, Cd2+ and Cr3+) also had little effect on the Na+ dependent L-Glu binding site. Cl--dependent L-Glu binding was potently inhibited by Na+ ions but was not affected by other monovalent ions. All of the divalent cations were potent inhibitors of both Cl--dependent and -independent binding. The results show that these binding sites of L-Glu can be distinguished by their response to cations and suggest possible novel modes of regulation in vivo. PMID- 2983844 TI - Synaptic long-term enhancement (LTE) induced by a heterosynaptic neural input. AB - The slow excitatory postsynaptic potential (s-EPSP) response of cat stellate ganglion can be enhanced for hours (long term enhancement, LTE) following a conditioning preganglionic train of low frequency pulses, even when test s-EPSPs are elicited by an unconditioned input line. This provides another model for associational alterations of synaptic efficacy. PMID- 2983845 TI - Comparison of group I non-reciprocal inhibition of individual motoneurones of a homogeneous population. AB - Investigation of differences in input unrelated to the type of motoneurones was performed using intracellular recording from soleus motoneurones. It has appeared that non-reciprocal inhibition of individual soleus motoneurones may be evoked from group I afferents of different muscles and via different interposed interneurones. PMID- 2983846 TI - [Therapeutic effects of prostaglandin E1 on experimentally induced atherosclerosis]. PMID- 2983847 TI - [Influence of epidural and general anesthesia on human plasma cAMP]. PMID- 2983848 TI - [ELISA in the detection of hepatitis A: detection of anti-HAV IgM with F(ab')]. PMID- 2983849 TI - Cross correlation studies in primate motor cortex: synaptic interaction and shared input. AB - Awake, unrestrained monkeys were trained to reach out with the forelimb and touch a button. Extracellular spike trains were recorded from pairs of neurons in contralateral precentral cortex with the same or separate microelectrodes. The neurons were located in the same or different functional columns as defined by intracortical microstimulation and passive sensory stimulation. Cross correlation analysis showed patterns consistent with synaptic excitation and/or inhibition between members of the cell pairs during the voluntary movement. The strength of correlation was inversely related to distance between columns, with the strongest correlations found between cells within the same column. Inhibitory correlations were virtually restricted to cell pairs within a single column. Temporal analysis showed that direct synaptic interaction and shared input patterns could be clearly distinguished in this physiologic setting. Spatial analysis indicated that shared input was concentrated among columns in the same and adjacent joint controlling zones as well as within a single column. No directional preference of shared input was present, a finding which was consistent with the observed nested organization of the forelimb area. PMID- 2983850 TI - delta-Opioid modulation of striatal dopaminergic activity in mice. AB - Mu- and delta-opioid subtype receptor antagonists were tested in the mouse for their effects on vertical climbing activity, an index of striatal dopaminergic activity. The selective delta-opioid antagonist ICI 154 129 (I/l) by itself enhanced vertical climbing activity in a dose-related manner, whereas the mu opioid antagonist naloxone by itself was inactive on climbing behavior. Naloxone increased the climbing-stimulant effect of I/l. Unstimulated vertical climbing activity was reduced, and all opioid-antagonist enhancement of climbing behavior was antagonized, by the competitive dopamine antagonist haloperidol in dose related fashion. The observed motor-enhancement effect of a selective delta opioid receptor antagonist is the first demonstration of physiologically significant tonic activity on a central opioid receptor. Our observations suggest that I/l may be useful clinically in striatal dopamine-deficient disease conditions such as parkinsonism. PMID- 2983851 TI - Adenoid cystic carcinoma: an unusual neurosurgical entity. AB - Adenoid cystic carcinoma, or cylindroma, usually presents in the salivary glands or in the upper respiratory passages. We report an unusual case of adenoid cystic carcinoma in a 45-year-old woman who presented with a three-year history of progressive right trigeminal sensory palsy, most pronounced in the second division. There was, in addition, a right trigeminal motor palsy and a partial right 6th nerve palsy. CT scanning showed only a small soft tissue mass spanning an enlarged right foramen ovale. Solid adenoid cystic carcinoma was resected from Meckel's cave via a middle cranial fossa approach. A subsequent biopsy of the right maxillary antral mucosa found tumor tissue. We concluded that the tumor originated in the maxillary antrum and spread posteriorly along the infraorbital nerve to enlarge in the foramen ovale. Radiation to a total of 5,000 cGy was given. At 22 months there was neither radiographic nor clinical evidence of recurrence. At 30 months, the development of unsteady gait signalled the presence of recurrent tumor extending backwards from Meckel's cave into the right cerebello-pontine angle. PMID- 2983852 TI - Risk factors in pediatric AIDS. PMID- 2983854 TI - The hematopoietic toxicity of regional radiation therapy. Correlations for combined modality therapy with systemic chemotherapy. AB - Using circulating granulocyte-monocyte precursor colony-forming units in culture (CFUc) numbers as a probe along with standard blood count (CBC), the authors have quantitatively examined the hematopoietic toxicity of conventionally fractionated radiation therapy (RT) when combined with concurrent systemic chemotherapy or when used alone. Among 20 patients with limited stage small cell lung cancer receiving systemic chemotherapy with cyclophosphamide, CCNU, and methotrexate, the addition of involved field chest RT resulted in increased hematopoietic toxicity as judged by increased need for platelet transfusion (P less than 0.05) and decreased frequency of measurable CFUc (P less than 0.04). Among 22 patients receiving regional radiotherapy alone consistent hematopoietic toxicity was also observed. This toxicity, although generally of only mild to moderate clinical significance, was detected earlier and to a greater degree in patients who required radiation to larger treatment volumes, who had significant amounts of bone marrow in the port, and who had a high percentage of cardiac output flowing through the port. These data suggest that the hematopoietic toxicity of regional radiotherapy may be additive to that of concurrent systemic chemotherapy and may occur more promptly and to a greater degree when treatment volumes are larger or incorporate increased amounts of marrow volume or cardiac output. PMID- 2983856 TI - Nephroblastomatosis and Wilms' tumor. Clinical experience and management of seven patients. AB - Nephroblastomatosis is an uncommon abnormality believed to represent the persistence of embryonal renal tissue. Its association with simultaneous or eventual Wilms' tumor has led to the speculation that it is a premalignant lesion. The accumulated clinical experience with this process and its progression to Wilms' tumor in three of seven individuals is discussed. It is concluded that conservative tissue sparing surgery, chemotherapy, and ultrasonographic follow-up are the approaches most appropriate for the management of these difficult patients. PMID- 2983855 TI - Combined methotrexate and high-dose vincristine chemotherapy with radiation therapy for small cell bronchogenic carcinoma. AB - The addition of methotrexate to a previously described regimen of cyclophosphamide, Adriamycin (doxorubicin), and high-dose vincristine (VAC) was tested in 50 evaluable patients with small cell bronchogenic carcinoma. Prophylactic whole brain radiation therapy was given during the first chemotherapy course and consolidation radiation therapy was given to the mediastinum and primary site after achieving partial or complete remission. The addition of methotrexate did not improve the incidence of complete remission as compared to a previous regimen without it. The addition of radiation therapy improved the local control rate. The high-dose vincristine in this and a previous CAV study improved the incidence of complete remission in both limited and extensive disease presentation as compared with the authors' previous experience and induced an acceptable and reversible neurotoxicity. Moderate dose consolidation radiotherapy to the lung primary and mediastinum was effective in improving local control. The distinction between limited and extensive disease was found to be vague, as 22% of the patients could be shifted from one group to the other depending on definition. The evaluation of the various staging procedures indicates that bone scan gave a small number of truly abnormal tests. Isotopic brain and liver-spleen scan could be duplicated by computerized axial tomography (CAT). CAT scan of abdomen disclosed unexpected extension to the retroperitoneal nodes and adrenals. It is concluded that radionuclide studies of brain, liver-spleen, and bone can be eliminated and can be replaced by CAT scan of brain, chest, and abdomen. Site of recurrence indicate that most intrathoracic recurrences took place outside the radiation therapy field and in the pleural space. An incidental finding was the high incidence of intramedullary spinal cord recurrence. PMID- 2983857 TI - Paget's disease of the nipple. Immunohistochemical localization of milk fat globule membrane antigens. AB - The authors have used the indirect immunoperoxidase technique to examine the presence and distribution of milk fat globule membrane antigens in 12 cases of mammary Paget's disease using two monoclonal antibodies, HMFG-1 and HMFG-2. These stain breast epithelial cells but do not stain normal epidermis. The Paget's cells showed a similar pattern of cytoplasmic staining to that seen in the underlying intraduct or invasive carcinoma, therefore confirming them to be malignant ductal cells. To support this one of the cases was stained with the antibody LE 61 which is specific for nonepidermal epithelial cytokeratins. The result was strongly positive in Paget's cells in the epidermis but not in squamous cells. PMID- 2983858 TI - Biochemical measure of the volume doubling time of human pulmonary neoplasms. AB - The volume doubling time (DT) of human lung neoplasms, determined from sequential, presurgery roentgenograms, was compared with biochemical and histologic observations on biopsy samples of the same tumors obtained during surgery. The DTs of the 16 neoplasms ranged from 24 days in an oat cell carcinoma to over 500 in a pulmonary carcinoid tumor, and showed a statistically significant, inverse correlation to the TK (thymidine kinase) and the UK (uridine kinase) concentration of the biopsy samples per g wet weight or mg DNA. Log DT bore a linear relationship to log TK (r = 0.75, P = 0.0008) and to log UK (r = 0.69, P = 0.0067), and an even better fit to the straight line was found when plotting the log of the standardized average of the two enzymes against log DT. The results demonstrate the feasibility of a biochemical method for determining the unknown DT of the many tumors that are not amenable (on account of shape, location, or lack of prior chest x-rays) to the direct, radiologic determination of this useful, dynamic parameter of clinical malignancy. PMID- 2983859 TI - Immunohistochemical demonstration of neuron-specific enolase in gynecologic malignant tumors. AB - Gynecologic malignant tumors were studied by the immunoperoxidase method for neuron-specific enolase (NSE). They included 22 argyrophil cell carcinomas of the endometrium, 6 argyrophil small cell carcinomas of the cervix, 21 argyrophil cell adenocarcinomas of the ovary (endometrioid type, 10; mucinous type, 11) and 3 ovarian carcinoids (strumal type, 2; insular type, 1). NSE was demonstrated in all cases of argyrophil small cell carcinomas of the cervix and ovarian carcinoids. On the other hand, NSE was positive only in four cases of endometrial carcinomas with argyrophil cells. Argyrophil cell adenocarcinomas of the cervix and the ovary were immunohistochemically negative for NSE. The current results suggest that argyrophil small cell carcinoma of the cervix, ovarian carcinoid, and some endometrial argyrophil cell carcinomas are related to APUDoma. PMID- 2983860 TI - Salivary gland malignancies in children. AB - Malignant salivary gland neoplasms in children are rare. Such tumors usually occur in the parotid gland of older children. Sixteen cases of major salivary gland malignancy are reviewed, 14 occurring in the parotid gland and two in the submandibular gland. Adenoid cystic carcinoma was the most common malignancy encountered, occurring in five patients; four have died of disease. Low-grade mucoepidermoid carcinoma occurred in three patients and all remain alive and free of disease. An assorted variety of other cancers occurred in eight patients. One child died of reticulum cell sarcoma of the parotid gland, and two children have died of adenocarcinoma. The remaining five patients remain alive and free of disease. Salivary gland malignancies in children appear to have similar biological activity as those occurring in adults. Such tumors must be treated according to the same principles generally accepted for management of salivary gland malignancies in adults. PMID- 2983853 TI - The management of status asthmaticus in infants and children. PMID- 2983861 TI - Human papillomavirus type 16 related DNA in an anaplastic carcinoma of the lung. AB - Twenty-four biopsy specimens from various histologic types of human carcinomas in the lung were analyzed for the presence of human papillomavirus (HPV) DNA. DNA from the individual specimens was tested for the presence of homologous sequences to HPV genotypes 1, 2, 4, 8, 9, 10, 11, 13, 16 and 18. One anaplastic carcinoma in the lung contained multiple copies of DNA hybridizing under stringent conditions to HPV 16 DNA. The latter DNA has been found to be frequently associated with human genital cancer (cervical, penile, and vulval cancer) and genital Bowen's disease. The HPV 16 positive lung tumor originated from a 61-year old female patient who underwent hysterectomy due to cervical cancer 9 years earlier. PMID- 2983862 TI - Effect of histologically verified TNM stage on disease control in treated small cell carcinoma of the lung. AB - Pretreatment classification of patients with small cell carcinoma of the lung into categories of "limited" and "extensive" disease is inadequate; it does not identify the few having a good prognosis for disease control. Available reports from the literature were analyzed for (1) histologically verified TNM stage; (2) adequate treatment by current standards; and (3) number and percent of patients remaining in complete remission at 30 months after the start of treatment. Long surviving patients by histologically verified stage were as follows: Stage I, 5 of 6 patients (83%); Stage II, 3 of 4 (75%); T3 without N2 or M1, 2 of 4 (50%); and N2 with any T value but without M1, 1 of 16 (6.2%). Long survivors with most distant involvement in the supraclavicular nodes were the following: ipsilateral, 3 of 22 (13.6%); contralateral, 2 of 40 (5%). Histologically verified M1 elsewhere allowed less than 1% long survivors. Indirect evidence of M1 by abnormal bone scan allowed less than 5%. Contrary to general usage, TNM staging of patients with small cell carcinoma of the lung promises to correlate closely with the probability of long disease-free survival. PMID- 2983863 TI - Nonhepatocytic malignant mixed tumor primary in the liver. Report of two cases. AB - Among primary tumors of the liver, the group classified as malignant hepatic mixed tumor or hepatoblastoma in adult is among the least common. In this report, two rare cases of nonhepatocytic malignant mixed tumor primary in the liver are reported. Diagnosis was established after autopsy in one and surgery in the other, who is still alive 2 years and 8 months after removal of the tumor by hepatic resection and may be the longest survivor among the nonhepatocytic malignant mixed tumor primary in the liver. PMID- 2983865 TI - Frequency, diagnosis, and prognosis of spinal cord compression in small cell bronchogenic carcinoma. A review of 817 consecutive patients. AB - A retrospective review of 817 consecutive patients with small cell bronchogenic carcinoma disclosed 29 cases of spinal cord compression (frequency, 3.5%). Twelve patients (group 1) presented with symptom of cord compression, whereas the remaining 17 patients (group 2) developed this complication during therapy. Clinical features differed in the two groups: pain was present in 83% of the patients in group 1 and in 47% of those in group 2. Pain preceded motor dysfunction in 75% and 12% of the two groups, respectively. Radiologic bone destruction of the spine was found in 11 of 12 patients in group 1 and in 3 of 16 patients in group 2. Myelography was performed in 8 patients in group 1 and in 14 patients from group 2. A complete block was found in seven and none of the patients in each of the two groups, respectively. Treatment with irradiation and/or laminectomy rarely resulted in a significant improvement of preexisting neurologic impairment but it frequently prevented the progression of symptoms, leading to preservation of sphincter control and ambulatory function in eight of nine and seven of eight patients, respectively. Careful evaluation is mandatory for patients presenting with back pain and radiographic evidence of bone destruction, as 11 of 26 such patients were found to have spinal cord compression. It is concluded that spinal cord compression presents in two distinct ways. One with early onset, pain, evidence of bone destruction, and complete myelographic block. The second is characterized by motor impairment with no evidence of bone destruction and only partial myelographic block. Treatment rarely ameliorates symptoms but prevents deterioration, making early diagnosis important. PMID- 2983864 TI - Fine-needle aspiration biopsy diagnosis of round cell malignant tumors of childhood. A combined light and electron microscopic approach. AB - A series of fine-needle aspiration biopsies (FNAB) from 76 children with round cell malignant tumors was studied by both light and electron microscopy. In 14 cases the material submitted for electron microscopy was inadequate, whereas in the remaining 62 biopsies a definitive diagnosis could be made. These included 29 malignant lymphomas, 8 neuroblastomas, 12 Ewing's sarcomas, 3 metastatic retinoblastomas, 3 Wilms' tumors, 3 rhabdomyosarcomas, and 4 unclassified tumors. The results from this study indicate that FNAB studied by light and electron microscopy can be effective in diagnosing small cell tumors in children. PMID- 2983866 TI - Antitumor activity of protein A administered intravenously to pet cats with leukemia or lymphosarcoma. AB - Extracorporeal immunoadsorption with protein A (SpA) containing Staphylococcus aureus Cowan I (SAC) has previously been shown to induce an antitumor and antiviral response in some feline leukemia virus (FeLV)-infected, lymphosarcoma (LSA) cats. However, the mechanism by which this response is induced is unknown. Since it is possible that SpA dissociates from the SAC column during treatment, and that intravenous infusion of SpA could be a more efficacious form of treatment than extracorporeal immunoadsorption therapy, 6 normal cats and 15 FeLV infected pet cats with naturally occurring leukemia or LSA were infused with SpA. No toxic effects resulting from SpA infusion were observed in the normal cats. Antitumor effects were observed in one of the eight cats with LSA and in three of the six cats with leukemia, but there were no antiviral responses. The antitumor responses were objective regressions; however, all tumors recurred. These results demonstrate that infusion of SpA into FeLV-infected LSA or leukemic cats can result in an antitumor but apparently not in an antiviral response. PMID- 2983867 TI - The insensitivity of 99mTc pertechnetate imaging for Kaposi's sarcoma. AB - A total of 13 technetium 99m (99mTc) pertechnetate image studies were performed on 7 patients with a histologic diagnosis of Kaposi's sarcoma. Five patients with peripheral edema had diffusely increased radionuclide activity in the same areas. The two patients without edema had normal radionuclide images. The primary cutaneous lesions were only identified in one of the patients. These findings suggest that previously reported "occult metastasis" are probably due to peripheral edema in most patients; therefore, the radionuclide study has a much lower sensitivity than has been previously reported. PMID- 2983868 TI - Characterization of immunoreactive forms of pancreatic polypeptide in islet cell tumors using antisera with different regional specificities. AB - Tumor tissue from a heterogeneous group of patients with clinically and histologically diagnosed islet cell tumors has been assessed for pancreatic polypeptide (PP) content using radioimmunoassay. Immunoreactive forms of PP, obtained by gel filtration of tissue extracts, were detected using antisera with different regional specificities. Normal pancreatic tissue contained one form of PP coeluting with human PP whereas, tumor tissue from patients with insulinoma, Zollinger-Ellison, and WDHA (watery diarrhea hypokalemic achlorhydric) syndromes contained, in addition, higher molecular weight forms of immunoreactive PP. Furthermore, a lower molecular weight form was detected in tumor tissue from a Zollinger-Ellison syndrome patient. The high molecular weight forms of immunoreactive PP were not recognized using a C-terminal-specific antiserum, whereas the smaller form was only detected using this antiserum. The variation in cross-reactivity observed with two antisera in this study emphasizes the limitations of using a single antiserum to detect molecular forms of peptide hormones, particularly those in islet cell tumors. PMID- 2983870 TI - Malignant rhabdoid tumor of the pelvis. AB - An extrarenal malignant rhabdoid tumor (MRT) of the pelvis in a 14-year-old girl is described. The location of the tumor and age of the patient appear to be unique. The neoplasm is histologically and ultrastructurally identical to malignant rhabdoid tumor of the kidney seen in infants and very young children. The tumor had an aggressive clinical course, and the patient died with disseminated disease 13 months after diagnosis. The authors review previous reports of extrarenal MRT, discuss the occurrence of cytoplasmic filamentous inclusions, and contrast MRT with other pelvic sarcomas. Although the exact histogenesis is unknown, ultrastructural and immunohistochemical data suggest a mesenchymal origin. PMID- 2983869 TI - Mucinous tumors of the ovary. A clinicopathologic study of 70 cases. AB - Seventy cases of mucinous ovarian tumor were reviewed. All patients were followed for a minimum of 5 years. Clinicopathologically, three groups were defined: (1) mucinous cystadenoma, which demonstrated no nuclear stratification and no stromal invasion (15 cases); (2) mucinous tumor of uncertain malignant potential, which was characterized by nuclear stratification of two to three layers and no stromal invasion (21 cases); and (3) mucinous carcinoma, which showed stromal invasion and/or nuclear stratification in excess of three layers (34 cases; 15 with invasion, 19 without). All patients with mucinous cystadenomas remained tumor free after initial surgery. Two patients with mucinous tumors of uncertain malignant potential died of tumor at 55 and 72 months, respectively, whereas 18 with mucinous carcinomas died after intervals ranging from 2 to 71 months. All mucinous tumors of uncertain malignant potential were Stage I at presentation. Twenty-one mucinous carcinomas were Stage I (six tumor deaths), one was Stage II (tumor death), ten were Stage III (nine tumor deaths), one was Stage IV (tumor death), and one was of uncertain stage (tumor death). Patients with mucinous carcinomas having stromal invasion demonstrated poorer survival (10 of 15 dead) than those with mucinous carcinomas lacking this finding (8 of 19 dead); however, stromal invasion was related to higher stage (5 with invasion Stage I; 16 without invasion Stage I). PMID- 2983871 TI - Bone metastasis in hepatocellular carcinoma. AB - Bone metastasis was observed in 16.1% or in 14 of 87 male autopsy cases of hepatocellular carcinoma. The primary tumor within the liver was located in the right lobe in all but one case. There were six patients who first presented with signs attributable to bone metastasis, and lung metastasis subsequently became evident in five of them. These 6 patients lived significantly longer as compared with 8 other patients with bone metastases and 73 patients without. The possible route by which hepatocellular carcinoma cells were carried to the bone is discussed. PMID- 2983872 TI - Testicular neoplasms occurring in four brothers. A search for a genetic predisposition. AB - Four brothers who developed testicular neoplasms, one bilaterally, are described. Histologic examination showed four of the tumors to be seminomas and one to be a mixed germ cell tumor. Three of the brothers are alive. Apart from a late-onset bladder carcinoma in their father and a pulmonary cancer in a maternal uncle, cancers were not recorded in the extended kindred. One patient, a sister, and the parents had normal frequency of sister chromatid exchange (SCE) and chromosome aberrations, whereas the two patients sampled after radiation showed increase in one or both. The father was found heterozygous in 12 and the mother in 8 genetic marker systems among 25 tested. For the blood group gene loci JK and MNSs, and the erythrocyte enzyme locus GPT the father had given the same allele to all three affected sons examined. The mother had given different alleles to the sons in all of her informative markers. On the model of a recessively acting susceptibility gene, only JK and GPT remained consistent with linkage without recombination. These investigations did not add support to a genetic etiology for the unusual family occurrence of testicular cancer. An apparent birth-order effect on time at onset/diagnosis in this and published families suggests time limited environmental factors. Nevertheless, JK, MNSs, and GPT should be included in future testis cancer families to test the model of a "dominant" genetic predisposition. PMID- 2983873 TI - Bronchogenic carcinoma in Olmsted County, 1935-1979. AB - The incidence rates of bronchogenic carcinoma based upon 414 cases that occurred in Olmsted County, Minnesota, over a period of 45 years are described. Histologic preparations were available for 97% of cases and were reviewed by one pathologist. The overall average annual age-adjusted rates per 100,000 person years for the most recent 5 years were 45.6 and 14.0 for men and women, respectively. Squamous cell carcinoma was most common among men and adenocarcinoma was the most common cell type among women. Incidence rates for women have risen to approximately those of men 30 years ago, whereas the incidence for men in the last 5 years has ceased to increase. Age-specific rates show increases for women in all age groups 55 years and older, and for men 65 years and older, whereas the incidence in men younger than 65 years has declined. Survival for 5 years is best for women for all cell types except small cell carcinoma, and is essentially unchanged from the earlier 40-year report. PMID- 2983874 TI - Sequencing of drugs and radiation. The integrated alternating regimen. AB - The association of radiotherapy (RT) and chemotherapy (CT) constitutes one of the main avenues for research in therapeutic oncology. This association has two aims: (1) increase in control rate of primary tumor (this requires either the potentiation of one of the two modalities by the other or the additivity of their effect on tumor cells without a parallel increase in the toxic effects on critical normal tissues); (2) spatial cooperation (RT being used for the control of the primary tumor or of the sanctuaries, and CT for the control of the disseminated disease). In these two strategies, RT and CT should be given up to full doses in order to be effective. The main risk is an increase in the number and severity of the early and late side effects. To circumvent this problem, two possibilities are being explored: (1) use of drugs without serious toxic effects on those critical tissues which are included in the irradiated volume; and (2) avoidance of concomitant administration and introduction of a sufficiently long time gap between the completion of one modality and initiation of the other. However, in such sequential treatment, a delay of CT until after the completion of RT, or an interruption of CT cycle during the course of RT, allows the occult metastases to increase in size; a similar delay in initiation of RT is also detrimental, as drugs are often not effective on bulky tumors. Moreover, under CT, the cells which are resistant to the cytotoxic drugs may disseminate and initiate chemoresistant metastases. Taking these disadvantages into account, a treatment protocol was proposed in 1980 in which CT and RT are given alternately, without undue delay. Chemotherapy is started with the usual scheduling of one cycle every month. Radiotherapy courses are interdigitated between CT cycles. Each course is initiated 1 week after interrupting CT and continued until 1 week before beginning the subsequent cycle of chemotherapy, and so on until completion of RT. Such split-course RT should have an effect on a tumor comparable to that of a conventional fractionation. This protocol has been used on 24 patients with non-Hodgkin's lymphoma (NHL) of diffuse histology, and 63 patients with small cell carcinoma of the lung. The 2-year relapse-free survivals are promising (in clinical stage II NHL of diffuse histology, 75%; and in small cell lung carcinomas, 33%).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2983875 TI - The role of radiation therapy in the treatment of small cell lung cancer. AB - Patients with small cell lung cancer (SCLC) are candidates for aggressive therapy because of their potential for long-term survival, especially patients with limited-stage disease. Although no treatment protocol can be considered "standard", the best results in limited-stage SCLC appear to be produced by a combination of chemotherapy and thoracic irradiation. Ongoing protocols testing the efficacy of thoracic irradiation should be able to settle question of the optimal treatment approach in limited-stage SCLC over the next 1 to 2 years. Careful attention to volume treated and the use of shrinking fields produce the best results with the minimum of toxicity. Treatment of extensive-stage SCLC has not been substantially improved to date with the addition of local or systemic irradiation. Prophylactic cranial irradiation reduces the incidence of CNS failure in SCLC and should be given, at a minimum, to patients achieving complete response status. Whether patients with partial response should also receive prophylactic cranial irradiation remains controversial. Finally, half-body radiation in SCLC is an experimental research technique that has shown some promise but remains quite toxic when combined with systemic chemotherapy. PMID- 2983876 TI - Sensitizers and protectors in radiotherapy. AB - The rationale for hypoxic cell radiosensitizers, of which misonidazole (MISO) is the most widely used drug, is based on the premise that hypoxic cells limit the cure rate of tumors to conventional radiotherapy. There is evidence that this is the case for tumors of the head and neck and possibly also for carcinoma of the cervix and lung. Despite numerous trials, however, MISO has not shown a significant clinical benefit. However, it can be demonstrated that little or no effect would have been expected at the doses of MISO used. New and more efficient radiosensitizers are now available which are more likely to produce significant improvements in local control with radiotherapy. The sulfhydryl radioprotector WR 2721 protects normal more than malignant tissues in animals from damage by radiation and chemotherapy agents. Evidence for protection of bone marrow and kidney has now been obtained for cyclophosphamide and cisplatin, respectively, in Phase I clinical trials. Sensitizers and protectors demonstrate that chemical modification of the therapeutic index is possible for both radiation and chemotherapy treatment of cancer. PMID- 2983877 TI - Particle radiation therapy. AB - Current interest in attempting to identify any therapeutic advantages of beams of heavy particles (heavier than electrons) over photons is based on differences in physical absorption and radiobiologic interactions. The article discusses: dose distributions in tissue, which are markedly different for particles than for high energy photons and so may be clinically advantageous for the former; differences in radiobiologic responses, which could lead to increased tumor cell killing and a possible increase in the therapeutic ratio for particles; clinical experience to date; directions for and impediments to future research. PMID- 2983878 TI - Radiation therapy for brain tumors. AB - Results of radiation therapy obtained at the University of California, San Francisco over the last 25 years for various adult types of brain tumors are presented. Included are astrocytomas, ependymomas, pineal and suprasellar tumors, meningiomas, and malignant gliomas. For each tumor type considered, the disease free survival rate appeared to be improved when subtotal resection was followed by irradiation. The lack of improvement in survival with malignant gliomas has prompted investigation into more aggressive multimodality therapies. These are discussed along with a new program using high-activity iodine 125 sources to deliver high-dose radiotherapy to malignant gliomas. It is possible that this new approach will lead to improved survival rates and be applicable to many tumors within the central nervous system. PMID- 2983880 TI - Histone kinase activities in normal and transformed mouse cells. AB - Crude extracts from replicating normal and transformed cells were assayed for protein kinase activities specific for different sites in purified Hl histone in vitro. Extracts from normal cells favored the NH2-terminal region while extracts from transformed cells favored the COOH-terminal region. Analysis of phosphopeptides demonstrated that histone kinases from both normal and transformed cells catalyzed the phosphorylation of a number of sites in common, and these were typical of sites phosphorylated in replicating cells. The preference for the NH2-terminal region by extracts from normal cells was due to the extensive phosphorylation of a site previously shown to be phosphorylated by cyclic AMP-dependent protein kinase. This activity was very low in transformed cells. PMID- 2983879 TI - Detection of lecithin: cholesterol acyltransferase (LCAT) in a human hepatoma cell line. AB - A human hepatoma cell line (HepG-2) was probed for the presence of lecithin: cholesterol acyltransferase (LCAT) using an antiserum to human plasma LCAT. Double immunodiffusion analysis using antiserum to human plasma LCAT revealed a single precipitin line in the sonicated cell homogenate. This precipitin line showed a reaction of identity with highly purified plasma LCAT. The presence of LCAT within the hepatoma cells was also confirmed by an immunofluorescence test. In contrast, the cell culture supernate showed a weak and inconsistent precipitin line. These data suggest that HepG-2 cells synthesize LCAT but secretion of the enzyme by these cells into the culture medium may be partially or totally impaired. PMID- 2983881 TI - Synthesis and study of spin-labeled nitrosoureas. AB - For the first time we have synthesized spin-labeled nitrosoureas and have studied their properties--reduction of the iminoxyl group by vitamin C leading to the formation of the corresponding hydroxylamine derivatives and degradation in the presence of an aminoradical, leading to biradicals. The ESR spectra of biradicals in methanol have nine hyperfine resonance lines. The spin-labeled nitrosoureas have shown a high antitumor activity against the L 1210 lymphoid leukemia and P 388 lymphocytic leukemia in BDF1 mice. A study of a broad range of transplantable tumors is in progress. PMID- 2983882 TI - Hereditary cancer, oncogenes, and antioncogenes. PMID- 2983883 TI - Regulation of melanin synthesis of B16 mouse melanoma cells by 1 alpha, 25 dihydroxyvitamin D3 and retinoic acid. AB - Melanin synthesis of B16 mouse melanoma cells was found to be stimulated dose and time dependently by 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], the hormonal form of vitamin D3. The stimulation of melanogenesis resulted from an increase in the activity of tyrosinase, a key enzyme in melanin synthesis. The minimum dose required for this stimulation was as low as 0.05 ng/ml, or 0.12 nM, a physiological level of plasma 1 alpha,25(OH)2D3. The stimulation by 1 alpha,25(OH)2D3 was specific; other derivatives of vitamin D3 caused no stimulation at a concentration of 500 ng/ml. When the cells were plated on agar plates, the proportion of dark or black colonies was not increased by the exposure to 1 alpha,25(OH)2D3. Furthermore, this compound did not induce melanin synthesis of an amelanotic variant. Thus, its stimulatory effect seemed to be due to stimulation of melanin synthesis of melanotic cells, rather than to conversion of amelanotic clones to melanotic ones. 1 alpha,25(OH)2D3 did not induce intracellular cyclic adenosine 3':5'-monophosphate, while cholera toxin induced cyclic adenosine 3':5'-monophosphate and stimulated melanin synthesis and tyrosinase activity much more than did 1 alpha,25(OH)2D3, suggesting that 1 alpha,25(OH)2D3 stimulates melanin synthesis by a cyclic adenosine 3':5' monophosphate-independent mechanism. B16 melanoma cells contained specific receptors for 1 alpha,25(OH)2D3. Scatchard plot analysis revealed two types of receptor; the high-affinity receptor had a Kd of 18.3 pM and an Nmax of 10.6 fmol/mg of protein. The specificity of receptor binding was demonstrated by studies showing that, for 50% displacement of 1 alpha,alpha,25(OH)2D3 binding, other derivatives were required at 500 times higher concentrations or more. In contrast to 1 alpha,25(OH)2D3, retinoic acid inhibited melanin synthesis and tyrosinase activity of B16 melanoma cells dose and time dependently. On simultaneous treatment, 1 alpha,25(OH)2D3 and retinoic acid caused mutual interference, and a balance between their respective stimulating and inhibitory effects was obtained at a molar ratio of 10:1; i.e., with 10 nM 1 alpha,25(OH)2D3 and 1 nM retinoic acid. PMID- 2983884 TI - Presence and regulation of tyrosinase activity in human neuroblastoma cell variants in vitro. AB - The human neuroblastoma cell line SK-N-SH comprises cells that undergo morphological and biochemical interconversion between a primitive sympathoblast and a variant, epithelial-like cell type which does not express the neuronal characteristics of the SK-N-SH cell line. Since neural crest cells, from which neuroblastomas are presumed to arise, can undergo transdifferentiation in culture from a neuronal phenotype into other cellular phenotypes, particularly into neurilemmal cells and melanocytes, the present study was undertaken to determine whether this capacity is preserved in malignant cells of the peripheral nervous system. Activities for tyrosinase, a melanocyte marker enzyme, and 2':3'-cyclic nucleotide phosphohydrolase, a Schwann-cell marker enzyme, were measured in clones of the two cell types. While no significant differences in 2':3'-cyclic nucleotide phosphohydrolase activity were measurable, tyrosinase activity was detectable only in the flattened neuroblastoma variant cell lines and was comparable to that in some human melanoma cell lines. The tyrosinase activity in neuroblastoma cell variants increased with cell density and was significantly elevated by melanocyte-stimulating hormone and 8-bromo-cyclic adenosine monophosphate, similar to that seen in melanoma cells in culture. Thus, our findings show that human neuroblastoma cells can undergo bidirectional transdifferentiation in vitro between a neuronal and a melanocyte phenotype, possibly reflecting a process which occurs in the patient. PMID- 2983885 TI - DNA damage and differential cytotoxicity produced in human cells by 2-chloroethyl (methylsulfonyl)methanesulfonate (NSC 338947), a new DNA-chloroethylating agent. AB - 2-Chloroethyl (methylsulfonyl)methanesulfonate (CIEtSoSo) is of interest as a possible new chloroethylating agent because its simpler chemistry suggests that it will not generate the hydroxyethyl products which are produced by chloroethylnitrosoureas (CIEtNUs). The effects of CIEtSoSo on DNA were studied in IMR-90 and VA-13 human embryo cells by means of DNA alkaline elution analysis. IMR-90 are normal cells, whereas VA-13 cells are SV40 transformed and are deficient in DNA-guanine O6-alkyltransferase activity. The effects of CIEtSoSo were essentially the same as those of CIEtNUs in the following respects: DNA interstrand cross-links were produced in VA-13 cells but not in IMR-90 cells; the interstrand cross-links in VA-13 cells were formed after a delay of 6 to 12 h and appeared to undergo repair; DNA-protein cross-links were formed promptly in both cell types and appeared to be repaired; DNA strand breaks and alkali-labile lesions were produced and repaired, and differences were observed between the two cell types; and VA-13 cells were more sensitive than IMR-90 cells (dose modification factor, 5). The differential cytotoxicity against VA-13 cells was similar to that produced by noncarbamoylating CIEtNUs, and significantly larger than that produced by carbamoylating CIEtNUs. The results suggest that CIEtSoSo acts by chloroethylating guanine O6 positions in DNA. PMID- 2983886 TI - Thyroid hormone modulation of transformation induced by Kirsten murine sarcoma virus. AB - We have investigated the effect of triiodothyronine (T3) on the transformation of normal rat kidney (NRK) cells by the Kirsten strain of murine sarcoma virus (Ki MSV). When NRK cells were grown and infected with Ki-MSV in medium lacking T3, the yield of transformed foci was about one-half that observed in the cultures supplemented with T3. Individual foci appeared somewhat later in cells grown out in medium devoid of T3. The yield of Ki-MSV released from transformed NRK cells was lower when these cells were maintained in T3-depleted medium. The results cannot be attributed to cell growth modification by T3. Normal and Ki-MSV transformed NRK cells grew equally well in mono-layer culture in medium containing or lacking T3. Selective maintenance and removal of T3 during various phases of the transformation process indicated that T3 exerted its maximum effect on transformation rates when added to the medium 24 h prior to virus infection. T3 was less effective in modulating transformation when added simultaneously with virus infection and was ineffective if added 24 h after virus infection. The results indicate that thyroid hormone is a required factor for optimal transformation by Ki-MSV and that the hormone exerts its effects during the early phase of Ki-MSV-induced transformation. PMID- 2983887 TI - Amplification of the c-myc oncogene in a subpopulation of human small cell lung cancer. AB - We have examined a panel of human lung cancer cell lines for amplification and expression of the c-myc, N-myc, and c-myb oncogenes. The cell lines analyzed represent various histopathological types of lung cancer: small cell carcinoma with neuroendocrine properties; squamous cell carcinoma with epithelial markers; and large cell carcinoma with a mixed neuroendocrine-epithelial phenotype. Two of six cell lines, both of which were small cell carcinomas, showed about a 20-fold amplification of the c-myc oncogene. In both cell lines, the amplification is accompanied by an enhanced expression of c-myc. The N-myc or c-myb genes were not amplified in any of the cell lines, nor were they expressed in detectable amounts. The results confirm and extend earlier findings on c-myc amplification in small cell lung cancer. PMID- 2983888 TI - Induction of epidermoid differentiation by cyclic adenine nucleotide in cultured mammary tumors of mice. AB - In search of the degree of responsiveness of mammary adenocarcinomas to signals of differentiation, mouse mammary tumors were induced to undergo a course of development leading to multiple foci of squamous metaplasia, and subsequently a differentiation manifested by marked keratinization. The mammary tumors had spontaneously arisen in the preneoplastic mammary outgrowths of the transplantable lines, D1, MH5, and MH9, after their long-term implantation in gland-free mammary fat pads of virgin BALB/c mice. The inductions were produced in cultured fragments of mammary tumors by incubation for 9 days in the cyclic adenine nucleotide, N6-O2'-dibutyryl cyclic AMP, at 0.1 mM, without or with prostaglandins E1, E2, and B1, each at 5 micrograms/ml, and 1 microM papaverine. The N6-O2'-dibutyryl cyclic AMP alone was as active in the mammary tumors derived from the D1 and MH9 preneoplastic outgrowths as was the entire mixture of inducers. Intracellular cyclic adenine nucleotide may presumably be the specific mediator of the inductive process, presumably being elevated synergistically by entry of the N6-O2'-dibutyryl cyclic AMP, by induction of adenyl cyclase by prostaglandin E1 and E2, and through inhibition of phosphodiesterases by papaverine. Epidermidalization occurred to equal extent in well-differentiated and anaplastic mammary adenocarcinomas, indicative that mammary tumor progression did not affect the susceptibility to this course of development and differentiation. Mammary gland epithelium retains its susceptibility to multifocal epidermidalization in organ culture throughout the gradient of neoplastic transformation and progression toward decreasing growth regulation, starting from normal mammary gland, next preneoplasia (both reported previously), then well-differentiated neoplasia, and lastly anaplastic cancer. The findings support the existence of a common or closely associated pool of progenitor cells for the alveolar and epidermoid courses of development and differentiation in mammary gland. Induction of squamous metaplasia and abundant keratinization in both the well-differentiated and anaplastic mammary adenocarcinomas caused some of the cells to differentiate terminally and to die. PMID- 2983889 TI - Induction of rat pancreatic B-cell tumors by the combined administration of streptozotocin or alloxan and poly(adenosine diphosphate ribose) synthetase inhibitors. AB - Streptozotocin and alloxan were administered to Wistar rats in combination with poly(adenosine diphosphate ribose) synthetase inhibitors. Ten to 16 months after the injection of streptozotocin (50 mg/kg body weight i.v.) and 3-aminobenzamide (345 mg/kg i.v.), streptozotocin (50 mg/kg) and nicotinamide (350 mg/kg i.p.), streptozotocin (50 mg/kg) and picolinamide (250 mg/kg i.p.), alloxan (40 mg/kg i.v.) and nicotinamide (350 mg/kg), alloxan (40 mg/kg) and 3-aminobenzamide (345 mg/kg), and alloxan (40 mg/kg) and picolinamide (250 mg/kg), pancreatic islet cell tumors developed in 100, 98, 60, 26, 22, and 20% of surviving rats, respectively. However, after the single injection of streptozotocin and alloxan, islet cell tumors developed in 42 and 11% of surviving rats, respectively. The tumors were rich in B-granules on electron micrographs and contained as large amounts of proinsulin messenger RNA as normal pancreatic islets. The results indicate that poly(adenosine diphosphate ribose) synthetase inhibitors enhance the tumorigenic effect of streptozotocin and alloxan on islet B-cells. PMID- 2983890 TI - Progabide: a new GABA-mimetic agent in clinical use. PMID- 2983891 TI - Immunosuppressant treatment in multiple sclerosis. PMID- 2983892 TI - Combined-modality treatment of inoperable lung cancer (i.v. immunotherapy, chemotherapy, and radiotherapy). AB - Patients with inoperable non-small cell carcinoma of the bronchus were treated by iv methanol extraction residue of bacillus Calmette-Guerin (MER) followed by combination chemotherapy with methotrexate, doxorubicin, cyclophosphamide, and lomustine (MACC). Radiotherapy was added in patients with localized disease. MER was given iv in four weekly courses. The dose ranged between 0.1 and 0.8 mg/m2/course and was determined according to the skin reactivity to MER. None of the 19 patients treated with iv MER had an objective tumor response during immunotherapy. Five of 16 evaluable patients receiving MACC (31%) achieved partial response. The overall median survival for all 17 patients treated was 11 months according to the protocol. During immunotherapy the following side effects were observed: all patients had fever and chills and most had malaise, nausea and/or vomiting, and headache. Transient abnormalities of liver function tests were found in six patients, and in one patient roentgenographic changes suggestive of lung granulomas were seen. The side effects were more severe during the first course than in the further treatments. Side effects of the MACC combination were comparable to those observed in previously reported studies. Immunological monitoring performed during immunotherapy revealed that iv MER had an unfavorable effect on the immune system. Following four courses there was a decrease in the in vivo skin reactivity to MER and to five recall antigens. Twenty-four hours after the initiation of iv MER there was a tendency for decrease in the lymphoproliferation to phytohemagglutinin, an increase in the indices of the adherent cell suppressor system and the prostaglandin-related suppressor system, and an increase in the percentage of adherent mononuclear cells. No consistent changes in serum lysozyme were found. A transient increase in the total number of peripheral blood leukocytes and a decrease in the number of lymphocytes were noticed 24 hours after iv MER. Taking into account the results of the immunological studies and the overall therapeutic results, we do not feel justified in continuing to use this combined modality treatment. PMID- 2983893 TI - Etoposide and very high dose cisplatin: salvage therapy for patients with advanced germ cell neoplasms. AB - Twelve patients with refractory germ cell tumors were treated with etoposide (VP16) (100 g/m2/day X 5) and very high dose cisplatin (VHD-CDDP) (40 mg/m2/day X 5) every 28 days. All patients had progressed or relapsed after therapy with vinblastine (0.15 mg/kg/day X 2), CDDP (20 mg/m2/day X 5), and bleomycin (30 units/week). Thirty-three cycles of VP16 plus VHD-CDDP were administered. Five patients achieved complete response (two with chemotherapy only, two with chemotherapy + surgery, and one with chemotherapy + irradiation), five achieved partial response (two continue therapy), and two had disease progression. Only one patient achieved prolonged complete response (24+ months). Myelosuppression (median wbc count nadir, 1200/mm3; median platelet count nadir, 18,500/mm3), neurotoxicity (five patients with diminished hearing and six with substantial peripheral neuropathy), and hypomagnesemia (21 of 22 courses; magnesium, less than 1.5 mg/dl) were the dominant toxic effects. No important nephrotoxicity was seen (median maximum creatinine, 1.2 mg/dl; range, 0.8-2.2). VP16 plus VHD-CDDP is safe and tolerable and may provide a non-cross-resistant alternative to vinblastine, CDDP, and bleomycin. PMID- 2983894 TI - Arterial drug infusion with extracorporeal removal. II. Internal carotid carmustine in the rhesus monkey. AB - During cancer chemotherapy by intra-arterial drug administration, systemic toxicity often limits the tolerable dose. We evaluated the pharmacokinetic advantage obtained by infusing carmustine (BCNU) into the internal carotid artery during BCNU removal from the blood from the perfused region by hemoperfusion. A hemoperfusion column (XR-010, Extracorporeal Medical Specialties) was shown to remove BCNU quantitatively from sheep blood flowing at 300 ml/minute when the drug was infused at 13 mg/minute for 30 minutes. Under general anesthesia, adult rhesus monkeys underwent catheterization of the internal carotid artery and placement of a catheter in the ipsilateral jugular vein at its junction with the sigmoid sinus. BCNU (10 mg/kg) was infused over 20 minutes while blood was pumped from the jugular vein through a small column and back into the inferior vena cava. The procedure reduced systemic exposure by 46%-84% compared with iv infusion of the same dose. Brain-to-systemic exposure ratios ranged from 18:1 to 87:1, depending on the pump flow rate and method of calculation. Hematopoietic toxicity was prevented. It is suggested that tumor exposure to BCNU comparable to that associated with very high tumor cell kill in vitro may be feasible with little or no systemic toxicity. PMID- 2983895 TI - Phase II evaluation of amsacrine in small cell lung cancer: a Southeastern Cancer Study Group trial. PMID- 2983897 TI - Immunohistochemical localization of cyclic AMP and ultrastructural demonstration of adenylate cyclase activity in the testis of Esox lucius at time of spermiation. AB - In the testis of Esox lucius at the time of spermiation, activity of cyclic adenosine 3',5'-monophosphate (cAMP) was immunocytochemically localized at the level of the Sertoli cells. In these cells adenylate cyclase activity was also ultracytochemically demonstrated by using adenylyl imidodiphosphate as a substrate. Reaction products of adenylate cyclase were primarily detectable on the basal and adluminal plasma membranes and on the surface of protrusions of the cell body into the lumen. PMID- 2983896 TI - Evidence for an association between calmodulin and membrane patches containing gonadotropin-releasing hormone--receptor complexes in cultured gonadotropes. AB - participation of calmodulin, clathrin, and actin in receptor mediated endocytosis of gonadotropin-releasing hormone (GnRH) was studied in an in vitro system of dispersed pituitary cells with a triple staining procedure. Cells were incubated in D-Lys6-Pro9-Des10-GnRH-biotin and stained with avidin-peroxidase diaminobenzidine. Calmodulin, clathrin, and actin as well as luteinizing hormone were identified by indirect immunofluorescence with FITC- and rhodamine-labeled second antibody. The results indicate a close spatial association of calmodulin, but not of clathrin and actin, with GnRH-containing plasma membrane patches. PMID- 2983898 TI - Response of isolated nuclei to phospholipid vesicles: a two dimensional gel electrophoresis analysis of H2SO4 soluble nuclear proteins after treatment with phosphatidylserine. AB - H2SO4 soluble proteins extracted from nuclei incubated with phosphatidylserine multilamellar vesicles (PS MLV) have been analysed by means of two-dimensional gel electrophoresis with particular attention to the uH2A fraction. A reduction of H1, H1 degrees and proteins A5, B7, B15 and B23 has been observed in lipid treated nuclei, while the core histones, as well as uH2A are unaffected by liposome treatment. Since these proteins show in vitro the same binding affinity for PS, their behaviour appears to be related to difference in localization in the nucleosome, responsible for their variable accessibility in the chromatin. These results might explain how this phospholipid induces a decondensation of chromatin and a stimulation of RNA synthesis. PMID- 2983899 TI - [Measurement of gastric mucosa and plasma cAMP and cGMP in patients with chronic gastric diseases due to splenic insufficiency and its clinical significance]. PMID- 2983900 TI - Dehydroepiandrosterone and 16 alpha-Br-epiandrosterone inhibit 12-O tetradecanoylphorbol-13-acetate stimulation of superoxide radical production by human polymorphonuclear leukocytes. AB - Both dehydroepiandrosterone (DHEA) and the synthetic steroid 16 alpha-Br epiandrosterone (Br-Epi), a more potent inhibitor of glucose-6-phosphate dehydrogenase (G6PDH) than DHEA, inhibit the 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulation of superoxide anion (O2-) formation by human neutrophils. DHEA has previously been shown to inhibit the development of spontaneous breast cancer and chemically induced tumors of the lung and colon as well as TPA promoted skin tumors in the mouse. The inhibition of TPA stimulated O2- formation by DHEA may contribute to the cancer preventive activity of this steroid. PMID- 2983901 TI - Reaction of DNA with a mutagenic 3-N,N-acetoxyacetylamino-4,6 dimethyldipyrido[1,2-a:3',2'- d]imidazole (N-AcO-AGlu-P-3) related to glutamic acid pyrolysates. AB - 3-Amino-4,6-dimethyldipyrido[1,2-a:3',2'-d]imidazole (Glu-P-3), an analog amine of the potent genotoxic Glu-P-1 isolated from a glutamic acid pyrolysate, has been chemically synthesized. Glu-P-3 was found much more mutagenic than Glu-P-1 to S. typhimurium TA 98 and TA 100 with S-9 mix. 3-N,N-acetoxyacetylamino-4,6 dimethyldipyrido[1,2-a:3',2'-d] imidazole (N-AcO-AGlu-P-3), a possible metabolite of Glu-P-3, binds covalently to the C-8 position of guanine residues in DNA. The binding induces large conformational changes of the macromolecule. PMID- 2983902 TI - Shock-induced damage to mitochondrial function and some cellular antioxidant mechanisms in humans. AB - The effects of circulatory shock on skeletal muscle mitochondrial oxidative activity in various substrates and cytochrome oxidase activity have been investigated using samples of muscle obtained by the needle biopsy technique from human subjects. The effect of shock on superoxide dismutase activity and glutathione content of skeletal muscle was also examined. The results show that there is a large decrease in cytochrome oxidase activity during shock and also in the capacity of the mitochondria to oxidize either succinate, or pyruvate, or palmitoyl carnitine. There is a fall in the tissue content of superoxide dismutase and in the total glutathione present. Furthermore, an increased oxidized glutathione content causes a decrease in the molar ratio of reduced to oxidized glutathione present in the muscle. These findings suggest that mitochondrial electron transport chain (ETC) oxidative damage can play a relevant role in the pathogenesis of circulatory shock and support the hypothesis of oxygen-free radical involvement in the cellular injury. PMID- 2983903 TI - Neuroendocrine and cardiovascular changes in septic shock and after cardiac surgery: effect of high-dose corticosteroid therapy. AB - Plasma levels of beta-endorphin-like immunoreactivity (BLI) were similarly elevated in patients with septic shock (group A) and in normotensive subjects recovering from cardiac surgery (group B) (1231 +/- 483 pg ml-1 and 1,240 +/- 355 pg ml-1, respectively). In neither group was cardiac output reduced, but total peripheral resistance index (TPRI) was low in group A and low or normal in group B. Intravenous methylprednisolone (MP) 30 mg kg-1 variably suppressed BLI by a mean of only 30% in group A, while in group B, BLI usually rose and then fell following MP. In group A percentage changes in BLI were positively correlated with percentage changes in cardiac index (CI) and mean arterial pressure (MAP) (r = .83, P less than .01, r = .59, P less than .05 respectively). No such correlations were found in group B. These findings suggest that increases in circulating beta-endorphin are unlikely to be responsible for myocardial depression or hypotension in septic shock. ACTH levels were in general normal in group A but were consistently elevated in group B, although plasma cortisol was similarly elevated in both groups. Furthermore there was a good correlation between percentage changes in ACTH and BLI following MP in group B (r = .87, P less than .01) but not in group A. Possible explanations for these findings are discussed. PMID- 2983904 TI - Effect of membrane depolarization on binding of [3H]nitrendipine to rat cardiac myocytes. AB - Binding of the dihydropyridine calcium antagonist [3H]nitrendipine was studied in cardiac myocytes incubated in normal and high potassium buffer so that we might examine the voltage dependence of dihydropyridine binding. Hearts were obtained from adult male Wistar rats, and isolated calcium tolerant myocytes were dissociated by enzymatic dispersion. Cells in 5.6 mM extracellular potassium showed specific binding of [3H]nitrendipine with KD 587 +/- 50 (mean +/- SE) pM and maximum receptor density, Bmax, 10.8 +/- 1.3 fmol/mg wet weight. Cells depolarized in 50 mM potassium showed no change in KD, 661 +/- 77 pM, but approximate doubling of Bmax 25.6 +/- 3.7 fmol/mg wet weight. Binding equilibrium was reached within 5 minutes at 37 degrees C, and the KD determined by kinetic analysis was in good agreement with KD determined by saturation experiments. Unlabeled nitrendipine and nifedipine completely inhibited [3H]nitrendipine binding with slope factors less than one, whereas verapamil and diltiazem only partially inhibited binding with slope factors substantially less than one. As a function of increasing extracellular potassium concentration from 2.4-54.1 mM, the number of nitrendipine-binding sites increased gradually 115%. Aconitine also produced a 58% increase in binding sites over a concentration range of 1-30 micrograms/ml. The potassium-induced changes in number of binding sites occurred rapidly and were rapidly reversible with changes in extracellular potassium concentration. There was no change in the number of [3H]nitrendipine-binding sites as a function of potassium or aconitine concentration in dead or digitonin treated myocytes. We conclude that nitrendipine receptor density in viable myocytes is voltage dependent, but we detect no change in KD as a function of voltage. PMID- 2983905 TI - Direct potentiometric determination of sodium ion in blood. III. Influence of (bi)carbonate. AB - We measured the emf of a sodium ion-selective electrode in NaCl-NaHCO3 solutions (160 mmol of Na+, 160-120 mmol of Cl-, and 0-40 mmol HCO3- per liter) with a home built cell in steady-state after 10 min and with two commercial direct potentiometric analyzers about 20 s after the sample was introduced. Substitution of HCO3- for Cl- resulted in a small decrease in emf. We calculated the effect of chloride ion replacement by bicarbonate ion on the liquid-junction potential and found that this accounted only for one-third of the emf decrease. The exact composition of bicarbonate solutions is closely related to the pH. To control the formation of carbonate, we performed measurements with the home-built cell at two pH values (7.0 and 8.2), controlled by tonometry with carbon dioxide. From the differences in the slope of the emf vs the amount of bicarbonate added at the two pHs we calculated the thermodynamic association constants of the sodium bicarbonate and sodium carbonate complexes (KNaHCO3(0) = 0.53 and KNaCO3- = 24). We conclude that, for the direct potentiometric measurement of sodium ion, only fresh serum should be used, to avoid pH changes. Replacement of sodium chloride by sodium bicarbonate caused different results when emf was measured with commercial analyzers, owing to their short measuring time of about 20 s. PMID- 2983906 TI - An experimental study of the effect of zinc on the activity of angiotensin converting enzyme in serum. AB - The activity in serum of zinc-dependent angiotensin converting enzyme (ACE), is measured to aid in diagnosis and monitor treatment of certain diseases. This report shows the effect of dietary zinc deprivation on ACE activity in the serum of rats. The mean (and SE) of the zinc concentration (mumol/L) in serum was 3.5 (0.3) in rats deprived of dietary zinc for four days, 16.3 (0.2) in control rats, and 19.8 (0.9) in rats deprived of zinc for four days, then repleted with zinc for 12 h. The respective mean (and SE) of ACE activities (nmol/mL per min) in serum were 390 (15), 543 (13), and 545 (20). Serum ACE activity was restored also by adding zinc to the assay mixture in vitro. The Vmax for ACE was 1.4 times greater when serum was diluted 40-fold as compared with twofold dilution. There was a small effect on the Km for the substrate, but the Km for zinc was decreased by 22-fold when serum was diluted 40-fold. The Vmax under these conditions was decreased by only 9%. PMID- 2983907 TI - A scheme for determining the correct activity of the kinetic angiotensin converting enzyme. AB - The absorbance difference measured when angiotensin-converting enzyme (EC 3.4.15.1) hydrolyzes the substrate N-[3-(2-furyl)acryloyl]-L phenylalanylglycylglycine is the basis for measuring its activity. We show this difference to be instrument dependent, and describe a method for deriving it that is applicable to manual or automated procedures. PMID- 2983908 TI - Pituitary ACTH dependent Cushing's syndrome due to ectopic production of a bombesin-like peptide by a medullary carcinoma of the thyroid. AB - A 41-year-old man presented with Cushing's syndrome and the biochemical features of ectopic ACTH production. Investigation revealed mediastinal metastases from a medullary carcinoma of the thyroid. The peripheral plasma contained grossly elevated levels of bombesin-like immunoreactivity (irBombesin) as well as calcitonin; blood sampling via a venous catheter confirmed a gradient of irBombesin, but not of ACTH, in the mediastinal vein draining the tumour. On extraction the tumour contained a bombesin-like peptide, but not vasopressin or corticotrophin releasing factor and only very low levels of ACTH; immunohistochemical studies showed positive immunostaining for bombesin and calcitonin but none for ACTH or CRF. No ACTH was released from dispersed tumour cells in vitro. However an extract of the tumour stimulated ACTH release in vitro from perifused dispersed rat anterior pituitary cells. This is the first reported case of Cushing's syndrome due to ectopic production of a bombesin-like peptide, causing excessive pituitary ACTH secretion. PMID- 2983909 TI - Effects of high-dose and low-dose naloxone on plasma ACTH in patients with ACTH hypersecretion. AB - The effect of a high (5.4 mg/h) and a low (0.8 mg/h) dose of naloxone (i.v. over a period of 90 min) on ACTH secretion was compared with placebo in patients with Addison's disease, congenital adrenal hyperplasia, Cushing's disease or Nelson's syndrome. In seven patients with primary adrenal insufficiency the high dose of naloxone provoked a significant increase of plasma ACTH concentrations (P less than 0.02) whereas the low dose of naloxone failed to influence ACTH secretion. In six patients with ACTH dependent Cushing's disease or Nelson's syndrome both doses failed to alter plasma ACTH levels. These results support the concept of inhibitory delta- or kappa-opiate receptors in the regulation of ACTH secretion. In patients with Cushing's disease or Nelson's syndrome ACTH secretion is insensitive to naloxone, presumably because of an autonomous pituitary adenoma or hypothalamic derangement. PMID- 2983910 TI - Effect of a single bolus of etomidate upon eight major corticosteroid hormones and plasma ACTH. AB - In a prospective controlled trial we investigated the effect of an induction dose of etomidate (0.26 mg/kg i.v.) on plasma ACTH, progesterone, 17 alpha OH progesterone, 11-deoxycortisol, cortisol, cortisone, corticosterone, 11 deoxycorticosterone, and aldosterone in seven males undergoing general anaesthesia. Seven other male patients receiving thiopentone at induction (5.0 mg/kg i.v.) served as controls. Plasma ACTH concentrations rose higher in the etomidate group (346 +/- 124 vs. 117 +/- 74 pg/ml, mean +/- SEM), but the difference was not significant. After etomidate we found a clear suppression of plasma cortisol (P less than 0.01), cortisone (P less than 0.01), corticosterone (P less than 0.01), and aldosterone (P less than 0.05) compared to corticosteroid levels after induction with thiopentone. Plasma 11-deoxycortisol and 11 deoxycorticosterone concentrations were grossly elevated 210 min after etomidate (91 +/- 28 nmol/l and 7.04 +/- 0.47 nmol/l, respectively, P less than 0.01) demonstrating inhibition of 11 beta-hydroxylation of both glucocorticoid and mineralocorticoid intermediates. In contrast, no significant difference in plasma progesterone and 17 alpha-OH-progesterone levels was found between the two groups indicating that the cholesterol-side-chain cleavage enzyme is less sensitive to etomidate than 11 beta-hydroxylase. Our results suggest that after induction of anaesthesia with a single bolus of etomidate, inhibition of other enzymes in the corticosteroid-synthetic pathway (e.g. cholesterol-side-chain cleavage enzyme) is of little clinical relevance. PMID- 2983912 TI - Cellular sensitivity to collagen in liver disease. AB - Peripheral mononuclear cells from 29 patients with liver disease and from 15 normal subjects were examined for cellular sensitivity to purified bovine type I and type II collagens by the in vitro production of leucocyte migration inhibitory factor. Mononuclear cells from 21 (69%) of the patients responded to type I collagen, but cells from only one (3%) did to type II collagen. In normal subjects, there were no response to both collagens. As compared with the corresponding humoral antibody response, the cellular sensitivity was more specific for type I collagen in patients with liver disease. These data demonstrate that cellular sensitivity to type I collagen exists in liver disease, presumably in relation to hepatic fibrosis. PMID- 2983911 TI - Enhanced expression in vivo of HLA-ABC antigens and beta 2-microglobulin on human lymphoid cells induced by human interferon-alpha in patients with lung cancer. Enhanced expression of class I major histocompatibility antigens prior to treatment. AB - The effect of cloned human interferon-alpha (IFN-alpha) on the expression of HLA ABC antigens (HLA-ABC) and beta 2-microglobulin (beta 2m) on human peripheral lymphoid cells in vivo was studied by cytofluorometry using monoclonal antibodies and fluorescein-labelled rabbit anti-mouse immunoglobulin. A significant increase in the mean fluorescence intensity of HLA-ABC (median 59%, P less than 0.001) and beta 2m (median 57%, P less than 0.001) on small lymphoid cells was observed 24 h after initiation of IFN-alpha treatment (50 X 10(6) units IFN-alpha/m2 three times a week). The enhanced expression of these antigens in vivo was found in 11 of 12 examined patients with primary bronchial carcinoma. A concomitant increase in serum beta 2m (median 90%, P less than 0.001) was found in all patients. In contrast the amount of cell-associated HLA-ABC and beta 2m remained unchanged (P greater than 0.1 and P greater than 0.5, respectively) by day-to-day analysis of an untreated healthy control group. An increased expression of both HLA-ABC (mean 55%, P less than 0.0005) and beta 2m (mean 23%, P less than 0.01) was also observed prior to treatment in the lung cancer patients when compared to a group of age matched healthy individuals. Treatment with IFN-alpha caused a significant redistribution of mononuclear cells resulting in both absolute and relative lymphopenia. Pre-treatment lymphocyte counts were 1.09 X 10(9)/1 (range 0.49 1.73), post-treatment counts were 0.55 X 10(9)/1 (range 0.39-1.06). PMID- 2983913 TI - Glomerular transplant rejection: a distinctive pattern of early graft damage. AB - Five hundred seventy-six consecutive biopsy or nephrectomy specimens obtained during the first 6 months of transplantation from 300 grafts in 431 recipients were examined by light microscopy for focal or diffuse endocapillary hypercellularity. Forty-seven (8.2%) of the 576 specimens obtained from 37 (12.3%) of the 300 grafts exhibited segmental or global occlusion of capillaries by swollen cells in 40-100% of glomeruli per biopsy. The lesions occurred at any time after transplantation, but 34 (72.3%) were present by day 60 and 7 (14.9%) before day 10. Immunofluorescence in 39 affected biopsies revealed focal or segmental glomerular staining in 18 (46.2%), among which IgM was found most frequently, and was considered to be non-specific. Electron micrographs of 17 biopsies from 14 grafts revealed that glomerular capillaries were narrowed or occluded by mononuclear cells of uncertain type, possibly monocytes, as well as lymphocytes and a few neutrophils. Complement-fixing antibody titers to cytomegalovirus rose at least fourfold in 10 (45.5%) of the 22 patients studied, but glomerular lesions were no more severe in the seroconverters than in the non converters, and there was no consistent temporal relationship between the occurrence of glomerular changes and seroconversion. Cellular or vascular rejection was present in most biopsies. One year graft survival was 34% among 35 accessed grafts with glomerular lesions, compared to 55% among 243 biopsied grafts with no glomerular changes. We consider that these lesions do not have a consistent association with cytomegalovirus infection and that they represent a distinctive form of glomerular rejection. Whether they indicate a poor graft survival, as the present results suggest but do not prove, requires further studies of other series of cases. PMID- 2983914 TI - Cytomegalovirus infection in dialysis patients. AB - We have studied cytomegalovirus (CMV) infection in 197 patients on regular dialysis treatment and 170 healthy platelet donors. Evidence of past CMV infection was found significantly more often in patients than in controls (137 of 197 v 60 of 170; p less than 0.001) at the commencement of the study. During a 12 month period 4 of the 60 dialysis patients initially found to be seronegative, but none of the 110 seronegative controls, developed primary CMV infection. Five of the 137 dialysis patients and one of the 60 controls initially found to be seropositive showed evidence of recurrent infection. Typically, there was only a transient elevation of CMV IgM antibody titer in both primary and recurrent infection. However, one dialysis patient with recurrent infection and another 5 initially seropositive patients showed persistence of CMV:IgM antibody production suggesting that they were experiencing chronic active infection. Neither primary nor recurrent infection was invariably a consequence of transfusion of blood or blood products. There were no clear-cut clinical sequelae from any of the three forms of infection documented. PMID- 2983915 TI - Mycobacterium haemophilum and mycobacterium xenopi associated infection in a renal transplant patient. AB - The case is presented of a renal-transplant patient in Europe with a Mycobacterium haemophilum infection in association with M. xenopi infection. Clinical signs suggested the diagnosis of mycobacteriosis, which was confirmed by a skin biopsy. Despite antitubercular treatment which rapidly eliminated M. xenopi, the patient's condition did not improve until M. haemophilum was identified. Minimal inhibitory concentrations of various antimicrobial compounds showed a lack of efficacy of isoniazid, and rifampin had no clinical effect. The patient recovered only after careful surgical drainage of the lesions and the administration of minocycline. The pathogenesis of such mycobacterioses is discussed, with focus on the immunodepressive status which in our patient may have been partially induced by a cytomegalovirus reinfection. PMID- 2983917 TI - Centronuclear myopathy with unusual mitochondrial abnormalities. AB - The case of a 34-years-old man is described with a progressive myopathy characterized by limb weakness and atrophy, involvement of facial, masticatory and extraocular muscles. The prominent features of the muscle biopsy were the presence of centrally located nuclei in most fibers. There was also an atrophy and predominance of type I fibers. Both clinical and morphological features were consistent with the diagnosis of centronuclear myopathy. Electron microscopic studies showed the presence of mitochondria with paracrystalline inclusions near the centralized nuclei but not in the subsarcolemmal position. This hitherto unreported feature led the authors to re-evaluate the hypothesis on the pathogenesis and the nosological classification of this myopathy. PMID- 2983916 TI - Primitive neuroectodermal tumors of the cauda equina in adults with no detectable primary intracranial neoplasm--three case studies. AB - We studied three cases of primitive neuroectodermal tumors surgically removed from the cauda equina region of adults. There was no clinical or radiological evidence of cerebellar medulloblastoma, other intracranial tumor or paraspinal sympathetic neuroblastoma. Two patients died: autopsies revealed no primary intracranial neoplasm. One patient is alive 3 years after surgery with no detectable intracranial tumor. The tumors had the light and electron microscopic features of primitive neuroectodermal tumors as described earlier in the literature. In addition, in all three cases many tumor cells could be stained for cytoplasmic neurofilament antigen. PMID- 2983919 TI - Membranous lipodystrophy (Nasu disease): clinical and neuropathological study of a case. AB - We report a sibling case of Nasu disease. A 35-year-old housewife, whose parents were consanguineous and whose sister died of the same disease, developed dementia, followed by bone fracture, incontinence and convulsions. She died at age 41. Pathologically, characteristic membranocystic changes of the fat cells in bone marrow and adipose tissues were observed. Neuropathologically, there was demyelination associated with intense gliosis and numerous axonal spheroids in the cerebral white matter. At the electron microscope level, these spheroids were an accumulation of various cell organelles. In addition, some had Hirano bodies. Incontinence was correlated with reduction of the number of nerve cells in Onuf's nucleus of the sacral cord. PMID- 2983918 TI - Intramedullary metastases: report of 4 cases and review of the literature. AB - Intramedullary metastases are very rare. We report four cases verified by autopsy. The site of the primary tumor was the lung (three cases) and the breast (one case). One case was complicated by a Brown-Sequard syndrome. We found 96 cases of intramedullary metastases in the literature. The primary tumor sites were lung (49%), breast (14%), kidney (6%) and colo-rectal region (5%). Melanoma was noted in 6% and Hodgkin's disease in 4% of the cases. Other primary tumors comprised 9%. The intramedullary metastases were most commonly thoracic. We compare our findings to those in the literature. PMID- 2983920 TI - Management of thumb duplication. AB - Radial polydactyly, a common congenital problem, most often occurs as thumb duplication. It appears in pentadactylous, polydactylous, and hypoplastic hands and is simulated by complex syndactyly. Classically, there are three categories for radial polydactyly: severe hypoplasia, partial duplication, and total duplication (easily confused with pseudoduplication). The common second category is subcategorized by Wassel's seven types. Management techniques vary with these classifications. Until a decade ago, the standard recommendation was simple excision of one of the duplicates. Problems of deformity, instability, and weakness became apparent, and reconstructive options were developed. Review of a series of 54 duplicate thumbs, all surgically treated, reflects the changing pattern of reconstruction. With current techniques, good results are usual, although secondary reconstructive procedures may be required. Simple excision is seldom indicated. Combination procedures, involving core tissues of bone, joint, and tendon or peripheral tissues--neurovascular, subcutaneous, and skin--or sometimes all of these, are preferred for reconstruction. The major current problem is to achieve the maximal good result with a minimal number of surgical operations. PMID- 2983921 TI - Congenital thumb deformities. AB - The congenital deformities that affect the thumb are reviewed in terms of the clinical problem, the type of inheritance involved, the associated abnormalities seen with the particular deformity, and the indications for surgical treatment. The thumb is often the key to establishing the diagnosis of many rare syndromes. The congenital thumb may be classified for easy identification by pediatricians, family doctors, and hand surgeons as follows: inadequate thumb, fingerlike thumb, contracted thumb, crooked thumb, extra thumb, and unusual thumb. PMID- 2983922 TI - Is ECT imaging with Tc(V)-99m dimercaptosuccinic acid useful to detect lung metastases of osteosarcoma? AB - ECT imaging, using Tc(V)-99m dimercaptosuccinic acid [Tc(V)-DMS] was performed in two patients with lung metastasis of osteosarcoma, and the results were compared with those of CT scan. Clear accumulation of Tc(V)-DMS was recognized in all cases in the same area that CT scans demonstrated. Tc(V)-DMS was labeled under optimal pH 8, had very low SnCl2 concentrations, an equilibrium between a stable form and a dissociated form of anion TcO4(3-) structurally similar to PO4(3-), and was postulated for tumor uptake. Considering this proposed mechanism for Tc(V)-DMS uptake by tumor cells, ECT imaging using this tracer could be of use in the early detection of lung metastasis of osteosarcoma. PMID- 2983923 TI - Persistent left superior vena cava detected with radionuclide angiocardiography. AB - A 28-year-old man presented with paresthesias, fatigue, central cyanosis, and erythrocytosis. A first pass flow study with Tc-99m as free pertechnetate was done, among other tests, to exclude a central shunt when a persistent left superior vena cava was incidentally detected. The value of radionuclide angiocardiography to examine the central circulation noninvasively was again illustrated in this case. PMID- 2983924 TI - 2,3-Dimercaptosuccinic acid as an antidote for lead intoxication. AB - 2,3-Dimercaptosuccinic acid (DMSA) is an orally effective drug more specific and with a wider therapeutic index than currently available drugs for lead intoxication. Eighteen men with elevated blood lead (BPb) concentrations received either 30, 20, or 10 mg/kg DMSA for 5 days in three divided daily doses. The mean BPb level decreased 72.5%, 58.3%, and 35.5% of the pretreatment values, with a simultaneous elevation in urinary Pb excretion. Clinical symptoms and biochemical indices of lead toxicity also improved. Red blood cell d-aminolevulinic acid dehydratase activity increased, while urinary excretion of d-aminolevulinic acid and coproporphyrin fell. DMSA was well tolerated; the only observed adverse drug reaction was a mild, transient elevation of serum SGPT levels in two subjects. DMSA appears promising and may greatly simplify the treatment of lead intoxication. PMID- 2983925 TI - Superoxide radicals, immunodeficiency and xanthine oxidase activity: man is not a mouse! AB - The importance of intact adenosine deaminase (ADA) activity in the generation of superoxide anion by xanthine oxidase has been disputed in studies using human neutrophils or mouse macrophages. The latter demonstrated a positive correlation between ADA activity and superoxide production during phagocytosis. The immunodeficiency in inherited ADA deficiency was related to a defect in this process. Since there is considerable interspecies variation in the tissue distribution of xanthine oxidase, the metabolism of [8-14C]deoxyadenosine (dAR), the toxic metabolite which accumulates in inherited ADA deficiency, was investigated in human peritoneal macrophages. Evaluation of the distribution of radiolabel in both cell and medium demonstrated that human macrophages with intact ADA metabolize dAR under physiological conditions to deoxyinosine and hypoxanthine exclusively. The hypoxanthine is further metabolized within the cell to ATP and GTP, via IMP. No xanthine or uric acid could be detected, confirming that in human macrophages xanthine oxidase activity is insignificant, as it is in most other human cells and tissues, except liver and intestinal mucosa. Thus production of superoxide radicals in such cells via this route would be impossible, and consequently unaffected either by ADA deficiency or the xanthine oxidase inhibitor allopurinol. PMID- 2983926 TI - Current indications for vasodilator therapy for left ventricular failure. PMID- 2983927 TI - Copy number of the chicken type II procollagen gene. AB - The copy number of the type II procollagen gene has been determined by quantitative hybridization of cloned DNA to Southern blots of total cellular DNA from chicken. Two different DNA probes have been used. One contains the 289 base pair exon coding for the junction between the triple helical domain and the carboxyl-terminal propeptide and the other contains an 162 base pair exon coding for amino acids 712-765 of the triple helical domain. Both fragments hybridize to single bands present at one copy per haploid genome in genomic DNA from chicken liver or sternal cartilage after these DNAs are cleaved with either HinfI of PstI. When the hybridization stringency and washing conditions were modified sufficiently to stabilize hybridization of heteroduplexes with up to 31% mismatch, no additional hybridizing fragments were detected. These results show that there is not a second copy of this gene or a pseudogene which is more closely related to the type II procollagen gene than the alpha 1 type I procollagen gene. These studies suggest that the cartilage 3 alpha collagen chain, which appears to be very similar to the alpha 1 (II) collagen chain, is encoded by the type II procollagen gene but is differently processed than the major type II gene product, and that the alpha 1 (II) Minor chain identified in bovine type II collagen preparations most likely represents an allelic form of type II collagen rather than a product of a different gene. PMID- 2983928 TI - Interaction of type I collagen with sepiolite (magnesium silicate). AB - Type I collagen from calf skin interacts with magnesium silicate (sepiolite) resulting in a collagen-clay complex which is separated by centrifugation. The interaction primarily occurs with high molecular weight aggregates of the protein as indicated by the fact that collagen from the skin of lathyritic rats interacts to a lesser extent. Thus, when calf skin collagen is fully retained, 45% of protein from lathyritic animals remains in solution. Monomeric forms of collagen remain soluble after short periods of interaction with sepiolite; at 5 minutes, 34% of the calf skin collagen preparation is recovered in apparently monomeric form. In contrast protein-protein interaction produces the complete retention of the total collagen with longer periods of time. PMID- 2983930 TI - Pustular reactions to hexafluorosilicate in foam rubber. AB - A case is reported of a foam rubber carrier with pustular lesions on the arms, wrists, thighs and trunk. Scratch and patch testing with the foam rubber components was negative. Animal testing revealed sodium hexafluorosilicate (Na2SiF6), one of the ingredients of the foam rubber, to be a pustulogen on previously damaged skin. PMID- 2983929 TI - Effects of oestradiol, testosterone and medroxyprogesterone on subcellular fraction marker enzyme activities from rat liver and brain. AB - The following enzymes have been studied (subcellular fractions are shown between parentheses): NAG and beta-glucuronidase (lysosomes); SDH (mitochondrial); glucose-6-phosphatase (endoplasmic reticulum); 5'-nucleotidase and (Na+, K+)Mg2+ ATPase (plasma membranes). Alterations on their activities were observed after subcutaneous injection of sex hormones, compared with controls. NAG activity from liver was always significantly decreased in lysosomal and microsomal fractions after the hormonal treatment. In the same conditions, NAG from brain was always increased. beta-Glucuronidase behaves like NAG in brain; in liver it was not modified by testosterone and it was slightly increased in lysosomal fraction after oestradiol treatment. SDH activity was not modified in mitochondrial fractions from liver, but this activity was always significantly increased in brain. Glucose-6-phosphatase activity was always significantly decreased in microsomal fractions from liver. It was increased in brain after oestradiol and testosterone injection, but medroxyprogesterone treatment caused a decreased activity. 5'-Nucleotidase and (Na+, K+)Mg2+ ATPase from brain were significantly increased in microsomal fractions by oestradiol and testosterone. Medroxyprogesterone, however, caused an increase in ATPase, but did not affect 5' nucleotidase. Both activities in liver were decreased by oestradiol and increased by testosterone, but medroxyprogesterone caused (Na+, K+)Mg2+ ATPase to rise and 5'-nucleotidase to fall. PMID- 2983931 TI - Receptors for T-cell growth factor: structure, function and expression on normal and neoplastic cells. PMID- 2983932 TI - The peripheral nervous system in dialyzed uremic patients: regressive motor unit changes. PMID- 2983933 TI - Ruminant nutrition: a century of progress. PMID- 2983934 TI - Advances in swine nutrition. PMID- 2983935 TI - Functional changes in mitochondrial properties as a result of their membrane cryodestruction. II. Influence of freezing and thawing on ATP complex activity of intact liver mitochondria. AB - The influence of the freeze-thawing rates on ATP synthetase (ATPase) complex of intact liver mitochondria was investigated. It was shown that the increase in latent ATPase activity and decrease in ATP synthetase activity resulted from an influence on the inner mitochondrial membrane. An increase in freeze-thawing rates led to the preservation of ATP synthetase activity and ATP hydrolysis reduction. Kinetic parameter changes of the ATP synthetase reaction resulted from an insignificant nonspecific increase in the inner mitochondrial membrane permeability and changes in its electrochemical potential level. PMID- 2983936 TI - Acquired immune deficiency syndrome: the causative agent and the evolving perspective. PMID- 2983937 TI - Human T-cell leukemia virus. PMID- 2983938 TI - Human T-cell leukemia/lymphoma viruses--an introduction. PMID- 2983939 TI - Adult T-cell leukemia/lymphoma and its clinical subtypes from the viewpoints of viral etiology. PMID- 2983940 TI - A retrovirus associated with a human leukemia, adult T-cell leukemia. PMID- 2983941 TI - Biology of T-cell leukemia virus: search for an animal system. PMID- 2983943 TI - Primary structure and processing of gag and env gene products of human T-cell leukemia viruses HTLV-ICR and HTLV-IATK. PMID- 2983942 TI - Some perspectives on the molecular mechanism of in vitro transformation and in vivo leukemogenesis by HTLV. PMID- 2983944 TI - Envelope properties of human T-cell leukemia viruses. PMID- 2983946 TI - Epidemiology of adult T-cell leukemia/lymphoma in Japan. PMID- 2983945 TI - Human T-cell leukemia/lymphoma viruses (HTLV): a unique family of pathogenic retroviruses. PMID- 2983947 TI - Human T-cell leukemia/lymphoma viruses: clinical and epidemiologic features. PMID- 2983949 TI - Clinical aspects of adult T-cell leukemia/lymphoma. PMID- 2983948 TI - Isolation, characterization, and biological effects of the first human retroviruses: the human T-lymphotropic retrovirus family. PMID- 2983950 TI - The epidemiology of human T-cell leukemia/lymphoma virus. PMID- 2983951 TI - Bilateral nephroblastoma associated with a 3;17 translocation. AB - Cultured cells from the tumor of a child with bilateral nephroblastoma were studied cytogenetically. All mitoses observed showed the same male karyotype, 46,XY,t(3;17). This translocation constitutes a newly discovered rearrangement that has not been reported previously either in nephroblastoma or in other neoplastic processes. PMID- 2983952 TI - Lung cancer in the young. AB - A long-term retrospective study was carried out on 1,514 cases of lung cancer to assess whether the disease presents substantial differences in young as compared to older patients. Clinical, epidemiologic, surgical, and survival data were evaluated in all cases. A young group under 45 years of age was studied separately and compared with the remaining older patients. In contrast with the literature, our results showed no percentage increase or variation in the male/female ratio of lung cancer in the young group. No significant difference was found regarding clinical picture, operability, histotype, and prognosis. PMID- 2983953 TI - [An epidemiological study of viral hepatitis A in Beijing suburb]. PMID- 2983954 TI - [Cyclic-nucleotides and adrenergic antiglaucoma drugs]. PMID- 2983955 TI - Single-cell analysis of the relationship among transferrin receptors, proliferation, and cell cycle phase in K562 cells. AB - Multiparameter single-cell analysis by flow cytometry was used to distinguish between size-related changes in K562 cell transferrin receptor (TfR) expression and changes in membrane receptor density throughout the cell cycle and over time in culture. Light-scatter pulse-width time-of-flight, a direct and readily calibrated measure of cell diameter, was used to calculate receptor density as the average number of receptors per unit cell surface area. Cell surface TfRs were unimodally distributed over the cell population and were present throughout the cell cycle. The number of receptors increased as cells progressed through the cell cycle, but cell cycle phase was also correlated with cell volume. However, when size heterogeneity was factored out by reanalysis of listmode data, there was a clear cell-cycle effect: among cells of the same size, both the number of receptors per cell and the receptor density increased from G1 to S to G2/M. TfR expression was also followed over time in culture after dilution into fresh medium. A decrease in growth rate after four days was preceded by one to two days by a decrease in both number of TfRs per cell and mean receptor density, indicating that decreased TfR expression represented true "down-regulation" and not just decreased cell size or an increase in the proportion of smaller G1 cells. This type of analysis is generally applicable for resolving the effects of cell size heterogeneity and cell cycle on membrane protein distribution and for other studies of ligand-receptor interaction. PMID- 2983958 TI - [Cell fusion and activation of Epstein-Barr virus (EBV)]. PMID- 2983956 TI - Cholesterol absorption and sterol balance in normal subjects receiving dietary fiber or ursodeoxycholic acid. AB - Twelve normal subjects were placed on a constant diet and evaluated during three treatment periods, each lasting four weeks. Patients received regular diet alone during period A, regular diet plus 60 g/day wheat bran during period B, and regular diet plus 15 mg/kg/day ursodeoxycholic acid during period C. Cholesterol absorption, plasma lipid concentration, daily fecal excretion of neutral and acidic sterols, and sterol balance were determined following each treatment. No changes in serum lipid levels are observed in the three study periods. Bran administration increases fecal neutral sterol excretion and decreases cholesterol absorption (from 50.1 to 42.0%) with respect to period C. Ursodeoxycholic acid administration has no effect on fecal neutral sterol excretion, whereas bile acid excretion is markedly increased. Sterol balance (evaluated as neutral and acidic fecal sterols excreted per day minus dietary cholesterol plus ursodeoxycholic acid given per day) is lower during ursodeoxycholic acid feeding (360 +/- 145 mg/day) than during bran feeding (593 +/- 174 mg/day). In conclusion, ursodeoxycholic acid decreases sterol balance values; bran-rich diet increases sterol balance and reduces cholesterol absorption. PMID- 2983957 TI - Inhibition of cytochrome P-450-dependent mixed function oxidation by ethanol. AB - The mechanism of inhibition of cytochrome P-450-dependent mixed function oxidation by ethanol was studied. Ethanol competitively inhibited the binding of hexobarbital to liver microsomes, and increased the low spin signal of cytochrome P-450 in the electron spin resonance spectra. Therefore, ethanol decreased the substrates bound to ferric cytochrome P-450 in the first step of mixed function oxidation. The second step of mixed function oxidation is the reduction of ferric cytochrome P-450-substrate complex by NADPH-cytochrome P-450 reductase. The NADPH dependent reduction of liver microsomal cytochrome P-450 was biphasic and composed of two first-order reactions. Ethanol decreased the rate constants of the fast and slow phases of microsomal cytochrome P-450 reduction. Thus, it is concluded that the inhibition of drug oxidation by ethanol may be due to the displacement of substrates from cytochrome P-450 and to the inhibition of reduction of cytochrome P-450 by NADPH-cytochrome P-450 reductase. PMID- 2983960 TI - [Transforming activity of DNA from primary hepatocellular carcinoma and the 7402 cell line on NIH/3T3 cells]. PMID- 2983959 TI - [Epstein-Barr virus nuclear antigen (EBNA) assay of the puncture smears from neck masses for the differential diagnosis of nasopharyngeal carcinoma]. PMID- 2983961 TI - [Cytogenetic study of the QP1 and QP2 lymphoblastoid cell line carrying EB virus genes in nasopharyngeal carcinoma]. PMID- 2983962 TI - [Metastatic ovarian carcinoma from a primary lesion in the gastrointestinal tract -an analysis of 20 cases]. PMID- 2983963 TI - [Relation between the growth pattern and the quantity of plasma cells of nasopharyngeal cancer (NPC) and serum EBV antibody level]. PMID- 2983964 TI - [AIDS and its relation to a virus infection]. PMID- 2983965 TI - [Varicella-zoster virus infection and the serologic determination of first infection immunity]. AB - The infection rate of varicella-zoster virus was determined by three tests- complement fixation reaction, enzyme-immune test and indirect immunofluorescence. The results of the three tests largely agreed with one another in demonstrating that infection starts relatively late in young children, but reaches 60%-70% at the end of the first decade of life, 90% at the end of the second decade. Results of both the enzyme immune and the indirect immunofluorescence tests indicated persistence of first-infection immunity until the older age groups. The complement fixation reaction deviated in its results, especially for the oldest age groups. PMID- 2983966 TI - [Specific gastrointestinal manifestations of a generalized cytomegalovirus infection in a patient with acquired immune deficiency syndrome]. AB - In a patient with AIDS, severe cytomegalovirus (CMV) infection dominated with gastro-intestinal and retinal involvement. The gastro-intestinal signs--diarrhoea and recurrent vomiting--correlated with the destruction of the tunica muscularis of the digestive tract seen at post-mortem. The CMV retinitis caused an almost complete loss of sight, the consequence of extensive destruction of the retina. In addition to the generalized CMV infection and disseminated Kaposi's sarcoma, which also involved the gastro-intestinal tract and caused discrete histopathological changes, post-mortem examination revealed a malignant lymphoma of the brain. PMID- 2983967 TI - [Clinical aspects and therapy of trophoblastic tumors]. AB - Over a period of ten years 14 women with trophoblast tumours (rarely seen in Europe) underwent diagnosis, treatment and follow-up, appropriate for the time. The high specificity of human beta-chorionic gonadotropin as marker in the recognition and follow-up of this tumour contributed to the good prognosis, as well as the cytostatic treatment, varied according to risk factors. Twelve of the 14 women are either cured or free of recurrences for a long time. PMID- 2983968 TI - [Acute and long-term effect of captopril in severe chronic heart failure]. AB - In nine patients with severe, treatment-resistant heart failure (stages IV in the NYHA classification) the acute and long-term effect of captopril were studied. In the acute experiment, peripheral resistance fell by 27% after administration of 25 mg captopril, cardiac index rose by 25%, arterial pressure, pulmonary arterial pressure and mean right atrial pressure fell by a similar amount. This haemodynamic improvement increased slightly in the course of longterm treatment (cardiac index +30%, peripheral resistance -30%, mean pulmonary arterial pressure -42%). The fall in heart rate by 15% and 25%, respectively, was an expression of haemodynamic improvement and reduction in angiotensin II. The fall in peripheral vascular resistance coincided with a 50% reduction in angiotensin II concentration. Over the longer term, 2-42 weeks, the renin system stimulation regressed with the improvement in haemodynamics. Four of the nine patients in stage IV improved to stage II, while the remaining five patients improved from IV to III. PMID- 2983969 TI - [Combination of dopamine and dobutamine in the differential therapy of acute and chronic myocardial failure]. PMID- 2983970 TI - [Spontaneous rupture of the liver caused by hepatoma]. PMID- 2983971 TI - [Structural and functional state of the erythrocyte membrane in experimental carcinogenesis]. AB - Structural and functional state of erythrocyte surface membrane was studied in C3H and C57B1/6 mice as well as in Wistar rats after 20-methylcholanthrene administration. It is shown that mice genetically susceptible to tumor growth as compared with mice resistant to carcinogenesis are characterized by an increased level of spinous erythrocytes (echinocytosis), higher membrane microviscosity and a tendency to increase polarity of their membrane lipid bilayer in the region of hydrophobic spine sound localization. The growth of chemically induced sarcomas was also accompanied by echinocytosis but with opposite changes in microviscosity and polarity of the erythrocyte membrane lipid bilayer. It is supposed that spinous erythrocyte transformation induced by different causes is based on different physicochemical factors. PMID- 2983972 TI - [Molecular genetic bases of the oncogenic action of the mouse mammary tumor virus]. AB - Processes associated with the neoplastic action of the mammary carcinoma virus on cells of the mammary gland myoepithelium are reviewed. Possible mechanisms of this action, stages of neoplastic process, at which the oncogenic MMTV potential is realized, and problems of virus and hormone cocarcinogenic action are discussed. PMID- 2983974 TI - [Comparative evaluation of the accumulation of 99mTc-labeled phosphate compounds in the bones]. AB - Distribution of osteotropic radiopharmaceuticals was studied in bone, different organs and tissues of rats. 99mTc-pyrophosphate (USSR and France) and 99mTc diphosphonate (GDR and Hungary) were investigated. Differences in their behaviour were demonstrated. According to a degree of accumulation in the bone tissue, the preparations may be arranged as follows: 99mTc-diphosphonate (GDR), 99mTc diphosphonate (Hungary), 99mTc-pyrophosphate (USSR and France). PMID- 2983973 TI - [In vitro determination of the sensitivity of lung tumors to different drugs]. AB - The in vitro sensitivity of 193 lung tumour specimens to anticancer agents was investigated using the short-term culture method. It is shown that there are many anticancer drugs with correlative effectiveness. It often concerns preparations with different mechanism of action. Using the in vitro sensitivity test data of correlative effectiveness it is possible not only to investigate the activity of new synthesized compounds and well known preparations but also to draw poly chemotherapy schemes with these agents in relatively short time. PMID- 2983975 TI - Evaluation of EMG examination as an indicator of worker susceptibility to organophosphates exposure. PMID- 2983976 TI - An assessment of the fibrogenic potential of very short 4T30 chrysotile by intratracheal instillation in rats. AB - Three groups of five rats each received, respectively, a single intratracheal instillation of saline (control), 5 mg of UICC chrysotile B asbestos, and 5 mg of a preparation of very short chrysotile fibers (4T30, 100% less than 8 micron) isolated by a sedimentation procedure. At various intervals after the treatment (1 to 60 days), assessment of lung morphology was performed on each animal. Although the two types of chrysotile fibers have similar chemical composition, structure, and surface charge, the lung tissue reaction differed considerably. Lungs of animals exposed to UICC chrysotile B showed significant pathological alterations as early as 7 days following treatment. The lesions were localized in and around terminal bronchioles and consisted of inflammatory cells, fibroblasts and collagen deposition which distorted and obstructed small airways. Reaction to very short 4T30 chrysotile fibers was quite distinct. Seven days after treatment, lungs of these animals showed alveolar and interstitial accumulation of inflammatory cells. The alveolitis persisted 60 days after treatment and no fibrosis was apparent. It appears that very short 4T30 chrysotile fibers are much less fibrogenic than UICC chrysotile B and that intratracheal instillations in rats may represent a useful mean of rapidly assessing the fibrogenic potential of various dusts. These observations support the concept that fiber length is an important factor for fibrogenicity of asbestos. PMID- 2983977 TI - The effect of mercuric acetate on selected enzymes of maternal and fetal hamsters at different gestational ages. AB - This study establishes levels of activity of glucose-6-phosphate dehydrogenase (G6PD), glycogen phosphorylase (GP), and cytochrome c oxidase (cyt c ox) in maternal, placental, and fetal tissues at Days 9, 12, and 15 in the 16-day gestation period of the hamster, and following a single dose of either 8 or 15 mg/kg mercuric acetate on the eighth gestational day. Mercury significantly elevated maternal kidney G6PD activity and decreased GP activity. The increase in kidney G6PD strongly correlated with observed urine and kidney abnormalities. PMID- 2983978 TI - Selective exposure and analysis of the sheep tracheal lobe as a model for toxicological studies of respirable particles. AB - A conscious sheep model, recently developed to study sequentially the bronchoalveolar milieu, was further refined to use in the rapid in vivo assessment of the biological effects of respirable particles. In this model, the anatomically isolated tracheal lobe was selectively exposed to either 100 ml phosphate buffered saline (PBS) (control group of 12 sheep), 100 mg of 0.1-micron latex beads in 100 ml PBS (latex group of 12 sheep), or 100 mg of UICC Canadian chrysotile fibers in 100 ml PBS (asbestos group of 12 sheep). Bronchoalveolar lavages (BAL) of the tracheal lobe were obtained prior to exposure and at Days 1, 8, 15, 21, 29, 45, and 60 after exposure. Whole-lung detailed pulmonary function tests (PFT) were performed at the same times and the histopathology of the lobe was examined in six sheep in each group at Days 29 and 60. In the control sheep, there were no significant changes over time in any of the measures, and lung morphology remained normal. In the latex group, there was no significant change in PFT, the BAL analyses documented early transient increase in cellularity (macrophages and neutrophils at Day 1) and only macrophages after; lung histology documented an early macrophagic alveolitis which decreased to less than 10% of the initial inflammatory reaction at Day 60, without other distortion of the lung and airway architecture. In the asbestos sheep, the only change in whole-lung PFT was a 10-torr fall in arterial O2 pressure. BAL analyses documented persistent increases in macrophages, neutrophils, and lactate dehydrogenase as well as increasing gamma-globulins. Lung histology revealed a macrophagic and neutrophilic peribronchiolar alveolitis at Day 30, which regressed substantially by Day 60, but persistent peribronchiolar alveolitis, early fibrosis, and severe distortion of the small airways, lesions comparable to those of early asbestosis in sheep or humans. Thus selective exposure and sequential analyses of the sheep tracheal lobe in terms of BAL histomorphology should be valuable for rapid in vivo assessment of toxicity of respirable particles. PMID- 2983979 TI - Lead silicate toxicity: a comparison among different compounds. AB - The aim of this report is to point out the relative toxicity of different commercial products containing lead silicates by studying their in vitro solubility and rate of absorption via the respiratory and digestive systems, compared with those of red lead (lead tetroxide). In vitro solubility was tested, by a modified Brimsdown method, with bidistilled water, saline, blood serum, pleural fluid, and dilute hydrochloric and acetic acids as solvents. Experimental animals were administered 25 mg equivalent mean doses using an intragastric tube or endotracheal intubation. Results showed that in vitro solubility gradually increases from distilled water, through saline and biological fluids, to acidic media. Lead absorption via the respiratory system is rapid and persistent, with small differences among the tested compounds. Less information can be gained from intragastric experiments. No correlation was found between in vitro solubility and in vivo absorption. On the other hand, in vitro studies must be strictly standardized. The relative absorption of the different materials, compared with red lead showed that all of them have to be regarded as equally toxic. PMID- 2983980 TI - Preparing for equine arteritis. PMID- 2983982 TI - Cytochrome c oxidase in plant mitochondria. Electron transfer after flash photolysis at low temperature of the CO-bound half-reduced complex. AB - Rebinding of CO after flash photolysis of the carboxy mixed-valence cytochrome c oxidase in plant mitochondria has been monitored in the visible range at low temperature (160-210K). At 586/630 nm, the photodissociation of the CO-saturated suspension was followed by a largely incomplete rebinding of CO to Fea3. After a series of three to four flashes, the unbound species accumulated up to 85% of the total enzyme, the remaining 15% CO-bound form undergoing recombination repetitively. In the Soret region, the absorbance level after the first flash came back to the initial one, as if all the CO-bound form were restored. However, the amount of complex which could be photodissociated by a second flash was only part of the starting material, in agreement with observations in the alpha range. The unbound species exhibited optical features very similar to the ones of compound C. The lack of CO rebinding is interpreted as due to an electron redistribution induced by the flash photolysis. This electron transfer has been tentatively assigned to a concomitant CuB oxidation and CuA reduction on the basis of the 655 nm band as a probe for a3+3 [Beinert et al. (1976) Biochim. Biophys. Acta 423, 339-355] and of the optical features in the visible range. Results are discussed with respect to analogous investigations with beef heart mitochondria and the studies by Boelens and Wever (1979-1982) of electron transfer after photolysis of the carboxy mixed-valence state when using the isolated mammalian enzyme at room temperature. PMID- 2983983 TI - Cap accessibility correlates with the initiation efficiency of alfalfa mosaic virus RNAs. AB - The rate of cap removal from the alfalfa mosaic virus (A1MV) RNAs with tobacco acid pyrophosphatase (TAP) depends on the RNA species. At 37 degrees C and in the absence of divalent cation, RNA 3 reacts more slowly than the other three, which are decapped at similar rates. In the presence of magnesium, at 25 degrees C, TAP also discriminates against RNA 1. Thus the order of reactivity with TAP largely mimics the hierarchy of initiation efficiencies of the A1MV RNAs (Godefroy Colburn et al., preceding paper in this journal). Our interpretation of these findings is that cap accessibility is what limits the rate of reaction with initiation factors as well as with TAP. In this hypothesis, translational discrimination between naturally capped messages would be related to the rate of 'breathing' of their 5' ends. PMID- 2983981 TI - DNA-binding properties of phosphorylated and dephosphorylated D2-T antigen, a simian-virus-40 T-antigen-related protein. AB - I have examined the role of phosphorylation of D2-T antigen in its DNA-binding properties and ATPase activity. Treatment of partially purified D2-T antigen with alkaline phosphatase resulted in removal of maximally 90% of the phosphate label associated with the radio-labeled protein. The specific and nonspecific DNA binding properties of partially dephosphorylated D2-T antigen were identical to those of the untreated control. In contrast, acid phosphatase was able to dephosphorylate D2-T antigen quantitatively. The general affinity for DNA of the completely dephosphorylated protein was unchanged or eventually slightly increased. However, its specific affinity for a restriction fragment containing the canonical T-antigen-binding sites was drastically reduced as shown by competition with unlabeled salmon sperm DNA. The results imply that nonspecific DNA binding of D2-T antigen is unaffected by phosphorylation whereas a specific phosphorylation site seems to be involved in the formation and/or stabilization of the specific protein-DNA complex. On the other hand, the ATPase activity of D2 T antigen seems to be unaffected by the degree of phosphorylation. PMID- 2983984 TI - Structure of the fructose-containing K52 capsular polysaccharide of uropathogenic Escherichia coli O4:K52:H-. AB - The chemical structure of the K52 antigenic capsular polysaccharide (K52 antigen) of Escherichia coli O4:K52:H- was elucidated by composition, nuclear magnetic resonance spectroscopy, methylation, periodate oxidation before and after graded acid hydrolysis and by oligosaccharide analysis. The polysaccharide consists of a backbone of alpha-galactose units interlinked between C1 and C3 by phosphodiester bridges. This poly(alpha-galactosyl-phosphate) is substituted at C2 of each galactose unit by beta-fructofuranose residues. About 80% of the galactose units are O-acetylated at C4 and about 10% of the fructose units are both O-acetylated and O-propionylated at C1. The K52 polysaccharide has an average molecular mass of 34 kDa, thus consisting of approximately 65 fructosyl-galactosyl-phosphate repeating units. PMID- 2983985 TI - Solubilization from rat pancreatic plasma membranes of a cholecystokinin (CCK) agonist-receptor complex interacting with guanine nucleotide regulatory proteins coexisting in the same macromolecular system. AB - Using the non-denaturing detergent 3-[(3-cholamidopropyl)dimethylammonio]-2 hydroxy-1-propane sulfonate (Chaps), cholecystokinin (CCK) receptors were solubilized from rat pancreatic membranes as a reversible complex with the CCK 31 39 nonapeptide 125I-labelled by the Bolton and Hunter reagent. Bound ligand dissociation from this soluble complex was similar to that from the membranous receptors of origin and the marked increase in the rate of dissociation induced by GTP was preserved in the soluble state, indicating that the solubilized CCK receptors remained functionally coupled with the guanine nucleotide regulatory site modulating the affinity for CCK. In fact, two guanine nucleotide regulatory proteins, Ns and Ni, coexisted in the soluble complex as established by identifying the 42-kDa subunit of Ns and the 40-kDa subunit of Ni, after ADP ribosylation by cholera toxin and Bordetella pertussis toxin, respectively. PMID- 2983986 TI - Preparation of highly purified F-actin-depolymerizing factor of human serum. AB - F-actin depolymerizing factor (ADF) of human serum was purified 250-400-fold to more than 98% purity with high reproducibility. The purification included (1) 30 50% (NH4)2SO4 precipitation, (2) ion-exchange chromatography on DEAE-Sepharose, (3) chromatofocusing on Polybuffer exchanger 94 and (4) affinity chromatography on ConA-Sepharose. The recovery of ADF was estimated to be 20-30% whereas the ADF activity yield was 5-17%. The lower activity yield was thought to be due-partly to proteolysis and partly to destabilization of highly purified ADF. PMID- 2983987 TI - A high-affinity site for glutathione in the cytoplasm of Escherichia coli and its possible role in potassium retention. AB - Glutathione-deficient mutants of Escherichia coli, unlike the wild type, exhibit a fast leak and a fast turnover of K+ at steady state. In a medium with low K+ the growth rate is very slow when unsupplemented with glutathione; when supplemented with glutathione at a concentration as low as 0.1 microM the growth rate is similar to that of the wild type. The ability of glutathione to restore a wild-type growth rate can be accounted for by an immediate reduction of the K+ leak. Two analogs of glutathione also reduce the K+ leak: gamma glutamylaminobutyrylglycine (ophthalmic acid) and alpha gamma glutamylcystinylbisglycine. Glutathione binds with high affinity (Kd 50 nM) to a cytoplasmic protein; ophthalmic acid and alpha gamma-glutamylcystinylbisglycine compete with glutathione for the binding site (Ki 0.1 microM and 1 microM), thereby ruling out the possibility that the thiol is involved both in reducing the K+ leak and in binding to the high affinity binding site. For each of the three peptides the Kd for binding is similar to the minimal concentration that achieves the maximal reduction of the K+ leak. The results suggest that the binding of glutathione should play a role in the retention of K+ in E. coli. PMID- 2983988 TI - Specific involvement of calmodulin and non-specific effect of tropomyosin in the sensitivity to ouabain of Na+,K+-ATPase in murine plasmocytoma cells. AB - The Kd for ouabain for inhibition of Na+,K+-ATPase isolated from murine plasmocytoma MOPC 173 cells is 120 microM, but when isolated in the presence of EDTA, it is 100-fold lower (1.2 microM). Simultaneous addition of muscle tropomyosin and calcium to sensitive membranes restored the original insensitivity (tropomyosin bound to the membranes in an irreversible and saturable manner). For comparison 86Rb influx into intact cells, mediated by the Na+,K+-pump, is half-maximally inhibited at 50 microM ouabain. Calcium converts the enzyme to an insensitive form. This appeared to involve calmodulin because after extraction of calmodulin with EDTA and EGTA from sensitive membranes, they could not be made insensitive by the addition of tropomyosin and Ca2+. Addition of exogenous calmodulin to these calmodulin-depleted membranes was required, in addition to tropomyosin and Ca2+, to decrease the ouabain sensitivity. The involvement of calmodulin was further assessed by measuring the range of Ca2+ concentrations required to convert to the insensitive form. At saturating concentrations of tropomyosin, increasing free [Ca2+] up to 3 microM led to an heterogeneous population of Na+,K+-ATPase forms. The calcium dependency was a saturable process. The shift to the insensitive form was half maximal at 0.65 + 0.11 microM free Ca2+ and was abolished by the addition of troponin I or trifluoroperazine (0.1 mM). These results suggest that, in murine plasmocytoma cells, the intrinsic sensitivity of Na+,K+-ATPase to ouabain might be regulated by a calmodulin-dependent process within a submembrane contractile-like environment. PMID- 2983989 TI - Phosphate dependency of phosphofructokinase 2. AB - Experiments performed at micromolar concentrations of inorganic phosphate support the conclusion that liver phosphofructokinase 2 would be completely inactive in the absence of inorganic phosphate or arsenate. The concentration of inorganic phosphate that allowed half-maximal activity decreased with increasing pH, being approximately 0.11 mM at pH 6.5 and 0.05 mM at pH 8. The effect of phosphate was to increase V and to decrease Km for fructose 6-phosphate, without affecting Km for ATP. Citrate and P-enolpyruvate inhibited the enzyme non-competitively with fructose 6-phosphate and independently of the concentration of inorganic phosphate. Phosphorylation of the enzyme by the catalytic subunit of cyclic-AMP dependent protein kinase did not markedly modify the phosphate requirement and its effect of inactivating phosphofructokinase 2 could not be counteracted by excess phosphate. A nearly complete phosphate dependency was also observed with phosphofructokinase 2 purified from Saccharomyces cerevisiae or from spinach leaves. By contrast, the fructose 2,6-bisphosphatase activity of the liver bifunctional enzyme was not dependent on the presence of inorganic phosphate. Phosphate increased this activity about threefold when measured in the absence of added fructose 6-phosphate and a half-maximal effect was reached at approximately 0.5 mM phosphate. Like glycerol phosphate, phosphate counteracted the inhibition of fructose 2,6-bisphosphatase by fructose 6-phosphate, but a much higher concentration of phosphate than of glycerol phosphate was required to reach this effect. PMID- 2983990 TI - Use of tyrosine-containing polymers to characterize the substrate specificity of insulin and other hormone-stimulated tyrosine kinases. AB - Synthetic copolymers containing tyrosine residues were used to characterize the substrate specificity of the insulin receptor kinase and compare it to tyrosine kinases stimulated by epidermal growth factor, insulin-like growth factor-1 and phorbol ester. In partially purified receptor preparations from eight different tissues insulin best stimulated (highest V) phosphorylation of a random copolymer composed of glutamic and tyrosine residues at a 4:1 ratio (Glu/Tyr, 4:1). The insulin-stimulated phosphorylation of this polymer was highly significant also in receptor preparations from fresh human monocytes, where insulin binding and autophosphorylation were difficult to detect. Other tyrosine-containing polymers Ala/Glu/Lys/Tyr (6:2:5:1) and Glu/Ala/Tyr (6:3:1) were also phosphorylated by the insulin-stimulated kinase but to a lower extent. A tyrosine kinase stimulated by insulin-like growth factor-1, and one stimulated by phorbol ester also best phosphorylated the polymer Glu/Tyr (4:1). The three kinases differed only in their capability to phosphorylate Glu/Ala/Tyr (6:3:1) or Ala/Glu/Lys/Tyr (6:2:5:1). Glu/Tyr (4:1) was a poor substrate for the epidermal growth factor receptor kinase which best phosphorylated the polymer Glu/Ala/Tyr (6:3:1). Three additional polymers: Glu/Tyr (1:1), Glu/Ala/Tyr (1:1:1), and Lys/Tyr (1:1) failed to serve as substrates for all four tyrosine kinases tested. Taken together these findings suggest that. Hormone-sensitive tyrosine kinases have similar yet distinct substrate specificity and are likely to phosphorylate their native substrates on tyrosines adjacent to acidic (glutamic) residues. Tyrosine containing polymer substrates are highly sensitive and convenient tools to study (hormone-sensitive) tyrosine kinases whose native substrates are unknown or present at low concentrations. PMID- 2983991 TI - Chemical modifications of the Na+-H+ antiport in Escherichia coli membrane vesicles. AB - The effects of chemical modifications of the Na+-H+ antiport in Escherichia coli have been analyzed by studying the resulting variations of the energy-dependent, downhill Na+ efflux from membrane vesicles. The histidyl reagent diethylpyrocarbonate (EtO)2C2O3 prevents the activation of the Na+ efflux mechanism by delta microH+ or its components. Inactivation of the antiporter by (EtO)2C2O3 is completely reversed by hydroxylamine. The data suggest that histidine residues are involved in the molecular mechanism of the Na+-H+ antiport. In contrast, no conclusive evidence suggesting participation of carboxylic, tyrosine or sulfhydryl residues in the Na+-H+ exchange reaction has been obtained. PMID- 2983992 TI - Enzymic synthesis of the 4Fe-4S clusters of Clostridium pasteurianum ferredoxin. AB - Ex novo enzymic synthesis of the two 4Fe-4S clusters of Clostridium pasteurianum ferredoxin has been achieved by incubation of the apoprotein with catalytic amounts of the sulfurtransferase rhodanese in the presence of thiosulfate, DL dihydrolipoate and ferric ammonium citrate. This enzymic reconstitution procedure was compared to a chemical one, in which the enzyme was replaced by sodium sulfide. A further comparison was made with the results previously obtained in the enzymic synthesis of the 2Fe-2S cluster of spinach ferredoxin, allowing the following conclusions to be drawn. The nature of the cluster to be inserted into the reconstituted iron-sulfur protein is determined by the apoprotein itself. The refolding of the structure of the iron-sulfur proteins around the newly inserted cluster is the rate-limiting step in both chemical and enzymic reconstitution. Rhodanese appears to play a role in the recovery of the native architecture of the reconstituted iron-sulfur protein(s). The extension to the 4Fe-4S centers of the rhodanese-based biosynthetic system allows this enzymic route to be proposed as a general way to the in vivo synthesis of iron-sulfur structures. PMID- 2983993 TI - Purification of GTP:alpha-D-mannose-1-phosphate guanyltransferase. AB - The enzyme GTP:alpha-D-mannose-1-phosphate guanylyltransferase from porcine thyroid tissue has been purified 69 900-fold on columns of blue-Sepharose, DEAE Sepharose, phenyl-Sepharose and agarose-GTP affinity materials. Although it exhibits a tendency to aggregate, the enzyme travelled, upon sucrose velocity sedimentation, as a single oligomer with a molecular mass of 412 kDa. Michaelis constants were determined to be 1.0 microM, 1.0 mM, 3.5 microM and 0.4 microM for GDP-alpha-D-mannose, pyrophosphate, GTP and mannose-1-phosphate, respectively. The enzyme appears to be specific for the mannose moiety but will accept an inosine replacement for guanine and a deoxyribose replacement for ribose in GTP. PMID- 2983994 TI - Collagenolytic cleavage products of collagen type I as chemoattractants for human dermal fibroblasts. AB - The chemoattractive properties of collagen in native (triple-helical) and denatured (random coil) conformation were compared in a Boyden chamber type assay to those of collagen fragments derived from cleavage with mammalian or bacterial collagenase using human embryonic dermal fibroblasts as target cells. Chemotaxis to native collagen required low collagen concentrations because fibril formation at high concentrations and at physiological pH and ionic strength prevented chemoattractiveness. Chemotaxis of denatured collagen was comparable to that of native collagen in solution. Cleavage of native collagen with mammalian collagenase increased, digestion with bacterial collagenase abolished its chemotactic activity. It is thought that these data may reflect the in vivo situation during inflammation and wound repair. PMID- 2983995 TI - Laminated cisternae of the rough endoplasmic reticulum induced by coronavirus MHV A59 infection. AB - The infection of murine fibroblasts of the sac- line with a coronavirus, mouse hepatitis virus strain A59 (MHV-A59), results in a novel modification to some cisternae of the rough endoplasmic reticulum (RER). From 8 hours post infection (h.p.i.) we see in thin sections pairs of cisternae closely, stably and uniformly aligned. Serial sectioning shows that the regions of pairing or lamination extend for many thousands of nm in two dimensions, with the spacing between the juxtaposed membranes remaining very uniform at about 18 nm. These structures appear coincident with the onset of accumulation of the viral glycoprotein E1 in the RER membrane but 2 hours after the viral glycoprotein E2 can first be detected there. Ribosomes are excluded from the paired cisternal surfaces, while budding of progeny virions has never been seen at the cisternal membranes facing the cytosol, although ribosomes bind there. The lumina of paired cixternae are usually devoid of virions which, however, accumulate in areas where the paired cisternae diverge. Electron immunocytochemistry shows that both E1 and E2 glycoproteins are abundant in the paired cisternae. Following labelling for the E1 glycoprotein we see a periodic fine structure, rows of "beads" with a centre to centre spacing of about 7.5 nm, in the region between the paired membranes. In oblique sections of this region in cells fixed as if for the immunoperoxidase labelling, but omitting all its steps we see parallel rows of "beads" separated by about 7 nm. We suggest that the membrane spanning viral glycoprotein E1 together with viral nucleocapsids may be involved in laminating cisternae of the RER. PMID- 2983997 TI - Radionuclide determination of right- and left-ventricular mixing. AB - Previous models for the dynamics of the transit of the bolus of activity of labeled red cells through the chambers of the heart have assumed the complete and instantaneous mixing of tracer with the cardiac blood pool. This assumption can be conveniently tested with a radionuclide procedure. We present a model of the bolus transit through the right and left ventricle that allows the quantitation of the mixing of labeled cells with the residual blood of the cardiac chambers. This model was used to analyze the time-activity curves obtained from a variety of patients undergoing first-pass dynamic scans of the heart. It is shown that technetium-labeled red blood cells and technetium pertechnetate have indistinguishable mixing properties, and the blood in the left ventricle is more thoroughly mixed than that in the right ventricle. PMID- 2983996 TI - Detection of different cellular sides in rat liver and kidney by two monoclonal antibodies raised against the nucleotide-sugar hydrolyzing enzymes phosphodiesterase I and CMP-sialic acid hydrolase. AB - Six different monoclonal antibodies were generated after immunization of mice with a partially purified enzyme preparation of rat liver, containing nucleotide sugar hydrolase (NSH) I and II. These enzymes are also known under the names phosphodiesterase I and CMP-sialic acid hydrolase respectively [11]. In the enzyme-immunoassay the antibodies directed against NSH I displayed some cross reactivity with the enzyme preparation of NSH II, and to a much lower extent the reverse was also true. Two antibodies, C and D highly reactive with NSH II and NSH I respectively, were used for immunocytochemical studies on sections of various rat tissues, which were known to contain high activities of both enzymes. Both antibodies were shown to be highly specific domain markers for different sides of the various cells. Antibody C was bound exclusively to the sinusoidal side of liver hepatocytes and to the basal side of cells from kidney tubule and epididymis. For antibody D the binding pattern was completely different, showing exclusive binding to the canalicular side of the hepatocytes and to the brush border membranes of kidney tubule cells, whereas in epididymis only binding to connective tissues was observed. Our studies clearly demonstrate, at least for liver and kidney, that NSH I and II are located at different cellular sides and that the monoclonal antibodies C and D can be used as domain markers for basal and apical sides of these cells respectively. PMID- 2983999 TI - Direct demonstration that the monoclonal antibody AMT-13 and interleukin 2 bind to the same molecule. AB - 125I-labeled surface molecules from mouse T lymphoblasts were fractionated by affinity supports coupled with recombinant interleukin 2 (IL2) and the monoclonal antibody (mAb) AMT-13. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated that two molecules of approximately 55 kDa and of approximately 180 kDa were bound in both cases. Sequential precipitation and SDS-PAGE analysis of the precipitated molecules revealed that only the approximately 55-kDa molecule eluted from AMT-13 mAb support was rebound to IL2 affinity support. In addition, IL2 inhibited specifically the binding of 125I labeled AMT-13 mAb to T lymphoblasts. Thus the results directly demonstrate that the a approximately 55-kDa cell surface molecule represents the IL2-binding protein and that the mAb AMT-13 reacts with this molecule. PMID- 2983998 TI - Autoradiographical investigations on the affinity of acetyl-(103Ru)-ruthenocene for the adrenal glands of mice. AB - Whole-body autoradiography was used to investigate the distribution of acetyl-103 ruthenium-(103Ru)-ruthenocene in female mice 2 and 24 h after intravenous application. Two hours after the application of acetyl-(103Ru)-ruthenocene, the nasal mucosa, colon, lung, liver, spleen and especially the adrenal glands were labelled. After 24 h, apart from the absence of labelling of the colon, the labelling pattern did not differ from that obtained 2 h after application. Again, the adrenal glands were particularly strongly labelled. Microautoradiography was performed to investigate the distribution of acetyl-(103Ru)-ruthenocene within the adrenal glands. It was shown that acetyl-(103Ru)-ruthenocene labelling was restricted to the zona reticularis and the inner zona fasciculata of the adrenal cortex. After the stimulation of glucocorticoid synthesis in the adrenal cortex by adrenocorticotropic hormone (ACTH) pretreatment, the labelled area in the zona fasciculata was clearly enlarged. It is concluded that acetyl-(103Ru)-ruthenocene has an affinity for those regions of the adrenal glands where androgen and glucocorticoid synthesis occur. PMID- 2984000 TI - Vagally mediated reflex and cardiac slowing induced by loperamide in rats. AB - The intravenous injection of loperamide induced an immediate fall in blood pressure and heart rate in anaesthetized rats. Both effects were inhibited by the opiate antagonists naloxone and MRZ 2266 BS. Bilateral vagotomy also inhibited both effects whereas atropine only reduced the bradycardia, but the combination of atropine and tertatolol suppressed the bradycardia. A high dose of loperamide induced bradycardia in pithed rats. This effect was prevented by MRZ 2266 BS but not by naloxone. It is concluded that loperamide can elicit a vagally mediated reflex involving vagal and sympathetic mechanisms and could stimulate cardiac opiate receptors, probably kappa, both effects leading to bradycardia. PMID- 2984002 TI - Prevention of supersensitivity-like phenomena in rat vas deferens by colchicine. AB - Rat vas deferens has been shown to become supersensitive to alpha-adrenergic agonists on brief treatment with epinephrine (Epi-treatment). Epi-treatment increased the contractile response and the number of alpha-adrenoceptors in the tissue. The application of colchicine (2.5 mM) during Epi-treatment prevented the supersensitivity-like phenomena and increase in number of alpha-adrenoceptors. Vinblastine (10 microM) also counteracted the effect of Epi-treatment but cytochalasin B (100 microM) and strychnine (2.5 mM) did not. It is suggested that the supersensitivity-like phenomena induced in rat vas deferens by Epi-treatment were due to a change in microtubular components of the membrane. PMID- 2984001 TI - P2-purinoceptors mediate both vasodilation (via the endothelium) and vasoconstriction of the isolated rat femoral artery. AB - The distribution of P1- and P2-purinoceptors in isolated rat femoral artery was studied by comparing responses to ATP, alpha, beta-methylene ATP and adenosine, both when endothelial cells were intact and when they had been removed by mechanical rubbing (as confirmed by histochemical staining and abolition of relaxations to acetylcholine). alpha, beta-Methylene ATP and ATP (but not adenosine or acetylcholine) contracted preparations at resting tone. alpha, beta Methylene ATP was significantly more potent than ATP. The potency of both alpha, beta-methylene ATP and ATP was significantly increased in the absence of the endothelium. These contractions were unaffected by tetrodotoxin, phentolamine and methysergide in concentrations sufficient to abolish contractions to perivascular nerve stimulation, noradrenaline or 5-hydroxytryptamine. Acetylcholine, ATP and adenosine relaxed arteries whose tone had been raised by 10(-6) M noradrenaline. Removal of the endothelium abolished relaxations to ATP (contractions were seen instead) and acetylcholine, but not to adenosine. alpha, beta-Methylene ATP further contracted the high tone preparation and was again more potent when the endothelium was absent. ATP and alpha, beta-methylene ATP were more potent as contractile agents when noradrenaline was present. These results confirm that endothelial cells can mediate vasodilation. They also show that ATP can act at P2 purinoceptors at two locations in the isolated rat femoral artery; one on the endothelium leading to vasodilation, and the other on the smooth muscle leading to vasoconstriction. P1-Purinoceptors, however, appear to be located on the smooth muscle only and mediate vasodilation. At the smooth muscle P2 purinoceptor, alpha, beta-methylene ATP is more potent than ATP, whereas at the endothelial P2-purinoceptor, the reverse is true. PMID- 2984003 TI - Measurements of 45Ca2+ uptake and contractile responses after activation of postsynaptic alpha 1-adrenoceptors in the isolated canine saphenous vein: effects of calcium entry blockade. AB - The increase in 45Ca2+ content produced by the EC100 concentrations of a series of alpha 1-agonists in the canine saphenous vein (CSV) was determined in the presence of 10(-7) M rauwolscine and found to be correlated (r = 0.92) with the intrinsic activities of the alpha 1-agonists in CSV. The degree of inhibition of 45Ca2+ uptake by nifedipine (1 microM) was inversely correlated (r = -0.97) with the intrinsic activities of the alpha 1-agonists and also inversely correlated (r = -0.95) with the 45Ca2+ uptake induced by the agonists. In the presence of 5 mM LaCl3, there was no significant 45Ca2+ uptake elicited by KCl (80 mM) or activation of postsynaptic alpha 1-adrenoceptors in this tissue, and the alpha 1 agonists were found to have a varied ability to release internal Ca2+ for contractions. It is concluded that (1) activation of alpha 1-adrenoceptors in CSV utilizes both intracellular and extracellular Ca2+ for contractions; (2) the increase in 45Ca2+ content after activation of post-synaptic alpha 1 adrenoceptors in CSV is directly correlated with the intrinsic ability of the alpha 1-agonists to induce contractions; and (3) the sensitivity of the 45Ca2+ uptake to nifedipine is inversely related to the intrinsic ability of the alpha 1 agonists to translocate extracellular Ca2+. PMID- 2984004 TI - alpha 2-Adrenoceptors in the HT 29 human colon adenocarcinoma cell line: characterization with [3H]clonidine; effects on cyclic AMP accumulation. AB - In the present work, [3H]clonidine was used to characterize alpha 2-adrenoceptors on the human adenocarcinoma cell line HT 29. The effects of alpha 2-adrenergic stimulation on cellular cyclic AMP levels were also investigated. The binding of [3H]clonidine on HT 29 cell membrane preparations was rapid and reversible. Scatchard analysis of the saturation curves indicated the existence of a single class of non-interacting sites with a KD of 1.29 +/- 0.07 nM and a Bmax of 114 +/ 7 fmol/mg of cell membrane protein. The binding sites for [3H]clonidine showed the required specificity for alpha 2-adrenoceptors. The potencies of alpha adrenergic compounds to displace [3H]clonidine binding ranked as follows: yohimbine greater than phentolamine much greater than prazosin for antagonists and clonidine greater than epinephrine greater than norepinephrine greater than phenylephrine much greater than amidephrine for agonists. When tested on intact cells, epinephrine, norepinephrine and clonidine were found to counteract, in a dose-dependent manner, the increase of cyclic AMP triggered by vasoactive intestinal peptide (VIP). Such inhibitory effects were abolished by the addition of yohimbine but not of prazosin. The physiological amines were the most efficient agonists: both epinephrine and norepinephrine inhibited VIP-induced cyclic AMP accumulation by 50-55% with KD values of 50 nM and 300 nM respectively. Clonidine was a partial agonist only, provoking a weak (25-30%) inhibition of VIP-induced cyclic AMP accumulation even at high concentrations. These results indicate that, like normal colocytes, human colon adenocarcinoma cells HT 29 possess alpha 2-adrenoceptors, the stimulation of which is associated with an inhibition of cyclic AMP production.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984005 TI - Modulation of hippocampal serotonin (5-HT) release by endogenous adenosine. AB - Slices of rabbit hippocampus were preincubated with [3H]serotonin then superfused continuously and stimulated twice electrically. The stimulation-evoked overflow of tritium was Ca2+-dependent, tetrodotoxin-sensitive and subject to modulation by serotonin autoreceptors. It was decreased by various adenosine receptor agonists in an order of potency that was typical for A1-(Ri-) receptors: N6 cyclohexyladenosine greater than (-)N6-phenylisopropyladenosine greater than 5'-N ethylcarboxamideadenosine greater than (+)N6-phenylisopropyladenosine = adenosine. The effects of the agonists were antagonized by 8-phenyltheophylline. The hypothesis that endogenous adenosine influences hippocampal serotonin release is supported by the following findings: both the adenosine receptor antagonists (theophylline or 8-phenyltheophylline (10 microM, each)) and the enzyme adenosine deaminase (10 micrograms/ml) increased, whereas R-E 244 (3 microM), an inhibitor of adenosine uptake, significantly decreased the evoked tritium overflow. When 8 phenyltheophylline was present throughout superfusion the effects of both R-E 244 and exogenous adenosine deaminase were abolished. It is concluded that serotonin release in the rabbit hippocampus is depressed by endogenous adenosine via A1-(Ri ) receptors. PMID- 2984006 TI - Autoradiographic localisation of opiate receptors in rat small intestine. AB - [3H]Naloxone and [3H]dihydromorphine are selective ligands for opiate receptors. Using an in vitro autoradiographic technique, binding of these ligands has been demonstrated to the villi and crypts in rat small intestine. These results indicate the presence of opiate receptor sites in the small intestine which suggests further a role for endogenous opiates in the transport functions of intestinal mucosa. PMID- 2984007 TI - The effects of progabide (SL 76002) on locomotor activity and conditioned place preference induced by d-amphetamine. AB - The effect of prior treatment with a GABA mimetic, SL 76002 (100 mg/kg i.p.), on amphetamine-induced locomotor activity and conditioned place preferences with amphetamine (1.5 mg/kg i.p.) was investigated. SL 76002 significantly attenuated the motor stimulant effects of amphetamine, without influencing the rewarding properties at least as determined by the place preference procedure. When injected alone, SL 76002 did not affect handling-induced locomotor activation, nor did it exhibit any aversive or reinforcing properties. The results suggest that separate neural systems may subserve the motor and rewarding properties of amphetamine. PMID- 2984008 TI - The effect of freezing and thawing or of detergent treatment on peripheral benzodiazepine binding: the possible existence of an endogenous ligand. PMID- 2984009 TI - Possible function of alpha 1-adrenoceptors in the CNS in anaesthetized and conscious animals. AB - The influence of St 587 (2-(2-chloro-5-trifluoromethylphenylimino)imidazolidine), a selective alpha 1-adrenoceptor agonist which easily penetrates the blood-brain barrier, was tested on behavior and cardiovascular functions, respectively. The substance (up to 10 mg/kg subcutaneously (s.c.)) did not increase the exploratory activity of naive mice. The hexobarbitone 'sleeping' time in mice was reduced in a dose-dependent manner (St 587 ED50 = 14.4 mg/kg s.c.). Haloperidol 10 mg/kg s.c. induced catalepsy which was antagonized by St 587 in a dose-dependent manner (ED50 = 2.7 mg/kg i.p.). Conversely, the alpha 1-adrenoceptor-blocking agents prazosin and corynanthine elicited catalepsy in mice which had been treated with a subthreshold dose (2 mg/kg s.c.) of haloperidol; the ED50 values of the antagonists were 0.26 and 4.7 mg/kg i.p., respectively. In anaesthetized cats blood pressure and heart rate were not affected by 100 micrograms/kg St 587 injected into the left vertebral artery. In conscious dogs with beta adrenoceptors blocked, the drug was without effect (100 micrograms/kg intracisternally) on vagally mediated reflex bradycardia, as evoked by intravenous noradrenaline injection. As a positive control the alpha 2 adrenoceptor agonist B-HT 920 which is equipotent to St 587 with respect to peripheral vasopressor effects in rats was injected with 10 micrograms/kg intracisternally and facilitated the reflex bradycardia. It is concluded that alpha 1-adrenoceptors within the brain mediate behavioral activation in states of CNS depression but remain without effect on cardiovascular centers. PMID- 2984010 TI - Opioid receptor binding characteristics of the non-equilibrium mu antagonist, beta-funaltrexamine (beta-FNA). AB - beta-Funaltrexamine (beta-FNA) bound to mouse brain membranes in a reversible and an irreversible (not removed by washing of the membrane) manner, and a portion of each type of binding was opioid-specific. Addition of 100 mM NaCl to the incubating medium enhanced the binding of beta-FNA to membranes. Using membranes preincubated with beta-FNA (1 microM) and then washed three times, the maximum number of binding sites available to [3H]morphine was markedly diminished whereas the affinity of morphine for binding sites was not significantly altered. The binding of [3H]naltrexone was also reduced markedly by beta-FNA pretreatment. In similarly pretreated membranes, the binding of [3H]methionine enkephalin [3H][D Ala2,D-Leu5]enkephalin (DADLE) or [3H]ethylketazocine was reduced to a smaller extent. Using brain membranes from mice pretreated with a single subcutaneous injection of beta-FNA (100 mg/kg) 48 h prior to use, the binding of [3H]methionine enkephalin was unaffected whereas the number of binding sites available to [3H]morphine was significantly reduced. The inhibition by various ligands of the reversible binding of [3H] beta-FNA resembled the relative ability of the same ligands to inhibit the binding of [3H]ethylketazocine. It was concluded that the irreversible portion of the binding of beta-FNA demonstrates a selectivity for mu over delta binding sites, and that the reversible portion of the binding of beta-FNA demonstrates a selectivity for kappa binding sites over mu or delta binding sites. As such, the binding characteristics of beta-FNA are consistent with its profile in vivo and in isolated tissue studies in vitro. PMID- 2984011 TI - A kappa-selective opioidergic pathway is involved in the reversal of a behavioural effect of adrenalectomy. AB - We have shown previously that adrenalectomized rats are immobile for only approximately 30% of a 5 min retest period, 24 h after an initial 15 min swimming exposure, compared with approximately 70% immobility for intact animals. The administration of ketocyclazocine, dynorphin-(1-17) or [Met5]enkephalin[Arg6,Phe7] immediately after initial exposure reversed the effect of adrenalectomy, whereas equimolar doses of morphine sulphate, [D-Ala2,D Leu5]enkephalin and dynorphin-(1-8) were inactive. MR2266, a kappa-selective partial agonist/predominant antagonist, did not reverse immobility but antagonized the reversing effect of ketocyclazocine. In conjunction with our previous studies, we interpret these data to show that one of the opioidergic pathways for the incorporation of information post-stress is kappa-selective. This kappa selectivity in turn suggests a possible physiological role for prodynorphin-derived peptides in memory. PMID- 2984012 TI - Human calcitonin increases both cyclic AMP and cyclic GMP accumulation in human kidney cells. AB - Salmon calcitonin stimulates both adenylate cyclase and guanylate cyclase systems in human kidney cortical cells but does not modify the cyclic nucleotide levels of medullary cells. In order to compare the effect of mammalian calcitonin and of calcitonin from the ultimobranchial body on cyclic nucleotides, the action of both salmon and human calcitonin was compared in intact kidney cells. The role of the regulatory unit in relation to the stimulation exerted by both calcitonins on the adenylate cyclase activity of kidney plasma membranes was also studied. Low concentrations of human calcitonin produced a significant increase of cyclic GMP accumulation in human kidney cortical cells. Higher hormone doses, active in stimulating the adenylate cyclase system, resulted in a progressive decline of the response. In kidney medullary cells human calcitonin was a more efficacious and potent stimulator of cyclic AMP than of cyclic GMP accumulation. Neither of the two calcitonins stimulated kidney cortical or medullary plasma membrane adenylate cyclase in the absence of guanylyl 5'-imidodiphosphate. However, in the presence of guanylyl 5'-imidodiphosphate, both calcitonins stimulated the cortical adenylate cyclase system. Under the same conditions, medullary adenylate cyclase activity was stimulated only by human calcitonin. These observations suggest that human calcitonin stimulates cyclic nucleotide accumulation in human kidney cortex and medulla, while salmon calcitonin is active only at the cortical level. This phenomenon could be explained on the basis of hormone-receptor binding. PMID- 2984013 TI - P1-purinoceptors mediate relaxation of the bovine bronchial artery. AB - Adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP), adenosine 5' monophosphate (AMP) and adenosine all relaxed the bronchial artery. All four purines tested were more efficacious than isoproterenol. Dipyridamole (10(-6) M) enhanced the relaxations due to adenosine and ATP while theophylline (10(-6) M) inhibited the relaxations due to adenosine and ATP. The results suggest the presence of P1-type purinoceptors in the bronchial artery of cattle. PMID- 2984014 TI - The mechanism of cGMP-induced relaxation in vascular smooth muscle. PMID- 2984015 TI - Inhibition of neuroeffector transmission in the rabbit mesenteric artery by [Met5]enkephalin. PMID- 2984016 TI - Effects of some atypical antidepressants on beta-adrenoceptor binding and adenylate cyclase activity in the rat forebrain. AB - The effects of daily administration to rats of desipramine, talsupram, tomoxetine, maprotiline, nomifensine, mianserin and citalopram (each 10 mg kg-1 day-1) for 4 weeks on [3H]dihydroalprenolol ([3H]DHA) binding in the cerebral cortex and on the noradrenaline-sensitive adenylate cyclase in the limbic forebrain were determined. Of these compounds, desipramine was alone in reducing [3H]DHA binding and in attenuating the cAMP response. Two selective noradrenaline uptake inhibitors, talsupram and tomoxetine each reduced the cAMP response but without affecting [3H]DHA binding. The other drugs lacked effect on both measures indicating (except for citalopram) that reduction in sensitivity of beta adrenoceptors and of the noradrenaline-sensitive cAMP response might not be a simple consequence of noradrenaline uptake inhibition. The lack of effect of citalopram on the sensitivity of the beta-adrenoceptor system suggests that antidepressants with selective 5-HT uptake inhibitory properties owe their antidepressant activity to other mechanisms. PMID- 2984017 TI - Effect of cyclic AMP on mesaconitine-induced analgesia in mice. AB - The effects of cyclic AMP, dibutyryl cyclic AMP, theophylline, isoproterenol and propranolol on mesaconitine (MA)-induced antinociception in mice were investigated employing the tail flick and acetic acid-induced writhing methods for the evaluation of antinociceptive activity. MA-induced antinociception was significantly potentiated by cyclic AMP and dibutyryl cyclic AMP. The phosphodiesterase inhibitor theophylline also significantly potentiated the MA induced antinociception. Further, MA-induced antinociception was markedly increased by a beta-adrenoceptor agonist, isoproterenol, and reduced by a beta adrenoceptor antagonist, propranolol. These results suggest that the antinociceptive action of MA is potentiated through cyclic AMP and stimulation of the central beta-adrenergic system. PMID- 2984018 TI - Interaction of [3H]arylazido aminopropionyl ATP ([3H]ANAPP3) with P2-purinergic receptors in the smooth muscle of the isolated guinea-pig vas deferens. AB - Following its photolysis in the presence of the isolated guinea-pig vas deferens, the ATP photoaffinity label ANAPP3 produces a specific antagonism of adenine nucleotide-induced contractile responses which are mediated by P2-purinergic receptors. To characterize the site of covalent photoincorporation of ANAPP3, intact vasa deferentia were treated with [3H]ANAPP3 and samples of homogenate, cytosol and a crude membrane fraction were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Photolysis of [3H]ANAPP3 (10(-5) M; 3.0 mu Ci/ml) resulted in the incorporation of radioactivity into cellular components with apparent molecular weights of 54-66 and 43-57 kilodaltons. The photoincorporation of [3H]ANAPP3 was associated with the crude membrane fraction and not the cytosol, was reduced in the presence of ATP in an ATP-concentration dependent manner, was lessened following pretreatment of the tissues with photolyzed nonradiolabeled ANAPP3, and was unaffected by the nucleoside transport inhibitor, dipyridamole. In tension studies on the same tissues the presence of ATP resulted in a concentration-dependent reduction in the initial contractile response to [3H]ANAPP3 the response to 3H was antagonized in tissues which had been pretreated with nonradiolabeled ANAPP3, and dipyridamole had no effect on the contractile response to [3H]ANAPP3. According to several criteria these findings indicate that the antagonism by photolyzed ANAPP3 of adenine nucleotide induced responses is a direct result of the covalent insertion at or near the recognition site of cell-surface P2-purinergic receptors. PMID- 2984019 TI - Are the toxicities of pentobarbital and ethanol mediated by the GABA benzodiazepine receptor-chloride ionophore complex? AB - Both barbiturates and ethanol have been reported to interact with the GABA benzodiazepine receptor-chloride ionophore 'supramolecular complex'. These observations raise the possibility that some of the pharmacologic actions of barbiturates and ethanol may be mediated through this complex. In this study we have administered a series of drugs which bind to various components of the complex in an attempt to antagonize the lethality of sodium pentobarbital, and ethanol-induced loss of righting reflex in mice. It was found that isopropylbicyclophosphate (IPPO), a cage convulsant which binds at or near the chloride ionophore, greatly reduces the overall mortality (and increases latency to death) of animals pretreated with a lethal dose of pentobarbital. Picrotoxin also decreases pentobarbital lethality, but only at doses which were usually lethal when given alone. Picrotoxin shortened, rather than increased, latency to death. Strychnine did not prevent pentobarbital lethality, suggesting that the IPPO effect is not shared by convulsants in general. IPPO did not prevent ketamine-induced deaths, which supports the notion that the protective actions of IPPO are specific for depressant drugs which act at the chloride ionophore. IPPO also significantly reduced the duration of loss of righting reflex induced by ethanol. These observations suggest that the use of compounds which have a high affinity for the chloride ionophore in vitro might be fruitful in developing a clinical treatment for barbiturate or ethanol toxicity. PMID- 2984020 TI - Altered binding properties of beta-adrenergic receptors and lack of coupling to adenylate cyclase in P815 mastocytoma cells. AB - P815, a murine mastocytoma cell line, possesses beta-adrenergic binding sites as assessed by using [3H]dihydroalprenolol (antagonist) and [3H]hydroxybenzylisoproterenol (agonist). The number of binding sites per cell was 29 000 for the agonist and 75 000 for the antagonist, as determined by direct binding assays and inhibition experiments on intact cells. On membrane preparations from the same cells, binding of alprenolol was only displaceable by antagonists, while stereospecific binding of hydroxybenzylisoproterenol was only displaceable by agonists. The P815 membranes also possessed an adenylate cyclase stimulated by Gpp(NH)p and NaF but not by 1-isoproterenol. The intracellular cAMP level of intact cells was not modulated by 1-isoproterenol or by 1-epinephrine, but was increased by forskolin. These results suggest that the beta-adrenergic receptor of P815 mastocytoma cells is non-functional. This may explain the failure of agonists to stimulate adenylate cyclase activity in these cells. PMID- 2984021 TI - Evaluation of the alpha 1- and alpha 2-adrenoceptor effects of detomidine, a novel veterinary sedative analgesic. AB - The in vitro receptor interactions of detomidine, a novel veterinary sedative analgesic, were studied. Detomidine caused a concentration-dependent inhibition of the twitch response in electrically stimulated mouse vas deferens with a pD2 value of 8.8. Clonidine and xylazine had the same effect with pD2 values of 8.7 and 7.5, respectively. The effect of detomidine was competitively antagonized by the alpha 2-blocking agents yohimbine, rauwolscine and idazoxan but not by the alpha 1-antagonists prazosin and corynanthine. The effect of detomidine was not antagonized by the opioidergic antagonist naloxone, the dopaminergic antagonist sulpiride, the serotonergic antagonist methysergide, the histamine H2-antagonist cimetidine, the histamine H1-antagonist diphenhydramine and the cholinergic muscarine antagonist atropine. Detomidine, as well as clonidine and xylazine, produced concentration-dependent contractions of rat anococcygeal muscle and rabbit aortic strips with pD2 values between 2.5 and 6.4. Intrinsic activities (compared to phenylephrine) varied between 0.5 and 0.7. The effects of detomidine in these two muscles could be antagonized by low concentrations of prazosin. In receptor binding experiments detomidine showed strong affinity to alpha 2 receptors. There was some binding affinity towards alpha 1-receptors also but only negligible or no affinity towards dopamine, opiate and adenosine receptors. In conclusion, the present results suggest that detomidine is a potent alpha 2 adrenoceptor agonist and that at high concentrations it can also stimulate alpha 1-adrenoceptors. PMID- 2984022 TI - Opiate receptor thermodynamics: agonist and antagonist binding. AB - The equilibrium thermodynamics of [3H]etorphine and [3H]diprenorphine binding to rat brain membranes were studied. In the absence of NaCl, the binding of [3H]etorphine was endothermic (delta Ho = +2.31 kcal/mol) and driven by a large increase in entropy (delta So = +51.8 e.u.). Under similar conditions, the binding of [3H]diprenorphine was exothermic (delta Ho = -2.78 kcal/mol). In the presence of 100 mM NaCl, [3H]etorphine binding was relatively isothermic (delta Ho = +0.61 kcal/mol) and driven by a large increase in entropy (delta So = +45.6 e.u.). NaCl significantly decreased both delta Ho (-2.78 to -7.48 kcal/mol) and delta So (33.0 to 18.5 e.u.) for [3H]diprenorphine binding. The data suggest that the agonist configuration of the opiate receptor exists in a more open and mobile (higher entropy) conformation than does the antagonist form of the receptor. PMID- 2984023 TI - Effect of urapidil on beta-adrenoceptors of rat atria. AB - The effects of the antihypertensive drug urapidil were studied on chronotropic responses to isoprenaline in rat isolated atria. Urapidil (1-30 microM) antagonized the responses to isoprenaline competitively, and the pA2 value was 6.05. Thus, in addition to its alpha 1-adrenoceptor blocking action for which the pA2 value is about 7, urapidil has beta-adrenoceptor blocking activity. PMID- 2984025 TI - ACTH-(1-24) antagonizes the contractile effect of morphine on the isolated rat colon. PMID- 2984024 TI - L-AMP-PCP, an ATP receptor agonist in guinea-pig bladder, is inactive on taenia coli. AB - L-Adenyl 5'-(beta, gamma-methylene)-diphosphonate (L-AMP-PCP), a potent ATP receptor agonist in the guinea-pig bladder, was tested on the guinea-pig taenia coli. L-AMP-PCP, unlike ATP, did not relax the taenia coli, and it neither enhanced nor inhibited the action of ATP. Unlike ATP, L-AMP-PCP was not degraded by ectonucleotidases on the taenia coli. The lack of pharmacological effect of L AMP-PCP on the taenia coli supports the suggestion that the ATP receptors here differ from those in the guinea-pig bladder. PMID- 2984026 TI - Activities of cAMP-dependent protein kinase and enzymes of 2',5'-oligoadenylate metabolism in NIH 3T3 cells deepening into the resting state. AB - The activity of cAMP-dependent protein kinase was found to increase continuously in the NIH 3T3 cells, deepening into the resting state. The increase correlated with intracellular level of heat-stable protein inhibitor of the protein kinase rather than with the cAMP content. The elevation of 2',5'-oligo(A) synthetase activity and the decrease in 2'-phosphodiesterase activity were also observed in the cells sinking into the resting state. The variations in enzyme activities were similar to those caused by the increase in the intracellular cAMP content described elsewhere. These results agree with the idea that the cAMP-dependent protein kinase is involved in the regulation of the enzymes of 2',5'-oligo(A) metabolism. PMID- 2984027 TI - Effects of cyclic AMP on limb bud chondrogenesis in low cell density culture. AB - In a previous paper, it was shown that the limb bud mesodermal cells differentiated into cartilage even at low cell density by lowering the serum content in the culture medium (Hattori & Ide, Exp cell res 150 (1984) 338) [20]. The present paper describes the effects of cAMP on limb bud chondrogenesis at low cell density. cAMP promoted chondrogenesis at low cell density in cultures with various concentrations of serum. The limb bud cells differentiated into cartilage cells without forming aggregates. cAMP inhibited the loss of chondrogenic capability in serum-rich medium. The relationship between cAMP level and serum content is also discussed. PMID- 2984028 TI - Studies on calmodulin-binding proteins (CaMBPs) in the cilia of Tetrahymena. AB - As a first step to elucidate the involvement of calmodulin in Ca2+-dependent regulation of ciliary motility, molecular species and properties of calmodulin binding proteins (CaMBPs) in Tetrahymena cilia were investigated by a modified [125I]calmodulin overlay method. At least 36 kinds of CaMBPs were detected. All the CaMBPs bound to calmodulin in Ca2+-dependent and calmodulin-specific manners, but they showed different Ca2+-dependencies. Several of CaMBPs bound to calmodulin in the presence of 100 microM trifluoperazine, several did in the presence of 8 M urea, and a few of them were highly sensitive to trypsin digestion. Among these CaMBPs, we noticed a 95 000-dalton (D) CaMBP present in the outerdoublet microtubule fraction, which possessed some attributes of the calmodulin counterpart suggested from the results of our previous paper [12]. We discussed a possibility that this protein might correspond to one of the protein components of the interdoublet link. PMID- 2984029 TI - Differentiation of U-937 human monocyte-like cell line by 1 alpha,25 dihydroxyvitamin D3 or by retinoic acid. Opposite effects on insulin receptors. AB - The monocyte-like human cell line U-937 has been differentiated in vitro by incubation with either 1 alpha,25-dihydroxyvitamin D3 or retinoic acid (RA) plus dibutyryl cyclic AMP (db-cAMP). Both methods were effective in inducing the appearance of maturation markers. Their actions on insulin receptors were the opposite, however; 1 alpha,25-dihydroxyvitamin D3 increased the binding of the hormone, while RA plus db-cAMP decreased the binding. These effects were specific for insulin, since the transferrin receptors were reduced by both methods of differentiation. Thus, the changes in insulin receptors during maturation in vitro depend on the inducing agent and are not causally related to the differentiation process. PMID- 2984030 TI - Transforming NRK cells with avian sarcoma virus reduces the extracellular Ca2+ requirement without affecting the calcicalmodulin requirement for the G1/S transition. AB - NRK rat cells infected with a transformation-defective, temperature-sensitive (ts) mutant of the avian sarcoma virus could not proliferate in Ca2+-deficient medium at a nonpermissive temperature (40 degrees C) that inactivated the viral pp60v-src-transforming product and rendered the cells phenotypically untransformed. However, these arrested cells were stimulated to initiate DNA replication with little or no delay while still in the Ca2+-deficient medium, either by adding Ca2+ or calmodulin at 40 degrees C or by reducing the temperature to 36 degrees C which restored the transformed phenotype by rapidly reactivating pp60v-src. The G1/S transition triggered by restoring the transformed phenotype was suppressed by three different anticalmodulin drugs (R24571, trifluoperazine, W7). The suppression by one of these drugs, trifluoperazine, was overcome by adding calmodulin. Thus, neoplastic transformation by the avian sarcoma virus sharply reduces the extracellular Ca2+ requirement for the initiation of DNA replication without bypassing a calcicalmodulin-dependent mechanism also needed for the G1/S transition. PMID- 2984031 TI - Characterization of beta-adrenergic receptors in fresh and primary cultured bovine corneal endothelium. AB - Homogenates of fresh bovine corneal endothelium and of cells from primary cultures exhibited high affinity binding of [3H]-dihydroalprenolol, a specific beta-adrenergic antagonist. The binding was rapid and reversible. Specific binding of the radioligand in each preparation was saturable with half-maximal binding occurring at 0.5 nM. Homogenates of fresh tissue consistently showed a higher maximal binding capacity than did those from cell cultures. Both homogenates bound adrenergic agents in a manner consistent with the labelling of beta-adrenergic receptors. The relative affinities of epinephrine and norepinephrine indicated that beta-adrenergic receptors in both preparations are of the beta 2 subtype. The beta-adrenergic agonist, isoproterenol, increased the cAMP content of intact, cultured endothelial cells 27-fold over control. This effect was completely blocked by the beta-adrenergic antagonist, propranolol. Bovine corneal endothelial cells in primary culture contain beta-adrenergic receptors which are essentially identical to those from in vivo sources and appear to be linked functionally to cAMP synthesis in these cells. PMID- 2984032 TI - Adrenergic receptor subtypes in rabbit iris-ciliary body membranes: classification by radioligand studies. AB - Receptor subclasses in iris-ciliary body cell membranes were determined by direct binding of radioactive dihydroalprenolol (DHA), yohimbine (YOH), WB-4101 WB) and prazosin (PRZ), classified respectively as beta 1 + beta 2, alpha 2, alpha 1, and alpha 1 subtype selective ligands (based on binding to brain adrenergic receptors). Specific binding was defined with appropriate unlabelled agonist and antagonist drugs in each case. Binding data were analysed by library programs of the PROPHET computer system. Subclass specificity was also determined indirectly by binding competition of the labelled ligand (at a three- to 10-fold Kd concentration) with increasing concentrations of 'cold' agonist or antagonist. Ligand binding parameters of unlabelled drugs were obtained from Dixon and Scatchard plots of the data. The adrenergic receptor density of iris-ciliary body is approx. 600 fmol mg-1 protein, of which 20-25% are primarily beta 2 receptors. Three distinct subpopulations of alpha-receptors, representing 10, 25 and 40-45% of the total, bind PRZ, WB and YOH, respectively, each with high specificity for its corresponding ligand but with 10- to 1000-fold lower specificity for the other two ligands. The majority of alpha-adrenergic receptors are of the alpha 2 subtype. A small population of receptors are similar to vascular postsynaptic alpha 1-receptors, while a larger subpopulation may have characteristics intermediate between alpha 1- and alpha 2-subtypes. PMID- 2984033 TI - Extracellular cytotoxicity by phagocytosing polymorphonuclear neutrophilic leukocytes: enhancement by a chemotactic stimulus. AB - This study investigated the influence of a chemotactic stimulus on the extracellular cytotoxicity mediated by phagocytosing polymorphonuclear neutrophilic leukocytes (PMN). We used N-formyl-methionyl-leucyl-phenylalanine (FMLP) as chemotactic peptide, opsonized zymosan as phagocytosable particle, and ox red blood cells (ORBC) as extracellular bystander targets. Phagocytosing PMN were found to kill ORBC efficiently, as determined by the 51Cr-release assay. FMLP, at the concentration of 100 nM, significantly enhanced the target cell lysis. PMN from two patients with chronic granulomatous disease and normal PMN plus catalase or free radical scavengers (mannitol, benzoate, histidine) were completely devoid of cytolytic activity both in the presence and in the absence of FMLP. The results indicate that the target cell lysis by phagocytosing PMN as well as the chemotactic peptide-related amplification of the lysis itself depend on the expression of the PMN oxidative cytotoxic potential. A similar response to a chemotactic stimulus in vivo could provide a mechanism for regulating PMN dependent cytotoxic and inflammatory processes. PMID- 2984034 TI - Bronchoalveolar lavage diagnosis of bronchiolo-alveolar carcinoma. AB - Differentiation between diffuse lung tumours and interstitial pulmonary diseases often requires open lung biopsy. We report two cases of bronchiolo-alveolar carcinoma presenting as diffuse pulmonary disease, in which the diagnosis was made by cytological examination of bronchoalveolar lavage. Bronchoalveolar lavage can, thus, differentiate between diffuse lung tumours and interstitial pulmonary diseases. PMID- 2984035 TI - Alterations of inhibitory processes in the dentate gyrus following kindling induced epilepsy. AB - In vitro hippocampal slices were prepared from control and commissural-kindled rats and the inhibitory processes of the dentate granule cells were assessed using paired perforant path stimulation. An early (20 ms) Cl(-)-dependent and a late (200 ms-8 s) Cl(-)-independent inhibition were shown to be present in the dentate. Enhancement of the late Cl(-)-insensitive inhibition was observed following the establishment of commissural kindling and persisted for periods of up to eight weeks following the last seizure. PMID- 2984036 TI - On the Purkinje cell activity increase induced by suppression of inferior olive activity. AB - Previous experiments performed in rats under barbiturate anaesthesia have shown a remarkable increase of simple spike firing rate in cerebellar Purkinje cells following inferior olive lesion or inactivation. The increase is due, at least in part, to the withdrawal of the tonic background activity of corticocerebellar interneurones, which have GABA as a chemical transmitter. Since barbiturates potentiate GABAergic inhibition, it is possible that the effect is due to the presence of this type of anaesthesia. In absence of general anaesthesia, we have compared the simple spike firing rate of the Purkinje cells in rats with intact inferior olive and 3-5 days after inferior olive lesion by 3-acetylpyridine. In the latter condition, the firing rate is significantly higher. In other rats, under urethane anaesthesia, which is not known to interfere with GABAergic transmission, the inferior olive has been reversibly inactivated by applying a cooling probe to the ventral surface of the medulla. Following cooling of the inferior olive on one side, a remarkable increase of simple spike activity, parallel to the disappearance of complex spike activity, has been observed in the Purkinje cells of the contralateral side. These results show that the presence of the simple spike firing increase, which follows the removal of the climbing fibre activity, does not depend on an anaesthetic which potentiates GABAergic transmission, although its amplitude is affected by the same anaesthetic. They suggest, therefore, that the tonic inhibition exerted by the olivocerebellar pathway on the Purkinje cells operates also in physiological conditions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984037 TI - Effects of metabolic inhibitors on evoked activity and the energy state of hippocampal slices superfused in vitro. AB - The effects of the metabolic inhibitors, arsenate (1,10 mM), iodoacetate (1 mM), alpha-cyano-4-hydroxycinnamate (alpha C4HC: 0.05, 0.15, 0.5 mM), malonate (10 mM) and 2,4-dinitrophenol (10 microM) on granule cell evoked activity and levels of energy metabolites of superfused hippocampal slices were investigated. Every inhibitor tested decreased the amplitudes of the population spikes, and also to a lesser extent, the rates of rise of EPSP. The effects were essentially reversible except in the case of iodoacetate. Concentrations of inhibitors, sufficient to depress evoked activity by at least 75%, did not significantly decrease tissue levels of phosphocreatine, ATP or total K+, with the exception of 1 mM iodoacetate. In slices superfused with 10 mM arsenate, 0.5 mM alpha C4HC, 10 mM malonate or 10 micron 2,4-dinitrophenol, the threshold EPSP for population spike generation was significantly higher than in inhibitor-free medium. These results are discussed in relation to the potential importance of non-oxidative glucose metabolism in maintaining evoked activity, and the possibility that during metabolic insults, tissue ATP utilisation is decreased to match reduced cytoplasmic ATP supply. PMID- 2984038 TI - Responses of cat motor cortex neurons to cortico-cortical and somatosensory inputs. AB - Intracellular techniques were used to investigate a cortico-cortical path from sensory cortex to motor cortex of cats. Cortico-cortical epsps were evoked in motor cortex neurons by microstimulation of area 3a. Epsps with latencies between 1.2 and 2.4 ms were identified as monosynaptic. These short latency cortico cortical effects were recorded in layers II through VI of the motor cortex. Neurons with monosynaptic cortico-cortical epsps also received excitatory inputs from forelimb nerves, usually from both muscle and cutaneous afferent fibers. The epsps evoked from forelimb nerves in motor cortex neurons were preceded by neural activity in somatosensory cortex. Time delays between arrival of inputs in sensory cortex and in motor cortex were compared to the latencies of cortico cortical epsps in the same motor cortex neurons. It was apparent that the timing was appropriate for the identified cortico-cortical path to have relayed some sensory inputs to motor cortex. PMID- 2984039 TI - Aminopyridine-sensitivity of spinal cord white matter studied in vitro. AB - Sections of dorsal columns of the spinal cord were removed from rats of various ages and studied in a sucrose gap chamber. The potassium channel blocking agent, 4-aminopyridine (4-AP), led to a pronounced increase in the area of the compound action potential of immature dorsal column axons. During the course of maturation this effect was attenuated but not lost. Occlusion experiments indicate that the 4-AP-elicited increase in area of the response was likely the result of multiple action potential discharge, an effect also present in dorsal root but not ventral root fibers (Bowe et al. 1985). These results indicate that the 4-AP-elicited changes in action potential characteristics previously described for sensory fibers in the peripheral nervous system are also present in the central nervous system extensions of these axons. PMID- 2984040 TI - Developmental changes in the cerebellar cortex after locus ceruleus lesion with 6 hydroxydopamine in the rat. AB - Three-day-old rats received an infusion of a small dose of 6-hydroxydopamine (6 OHDA) (12 micrograms) directly into the locus ceruleus nuclei, in order to induce a subtotal lesion of the noradrenergic system. Significant modifications in cerebellum maturation were observed, mainly a regression failure in the perisomatic Purkinje cell processes in the depth of the fissura prima of 10-day old rats, a reduction in the whole vermis surface area and in the total number of internal granule cells at 15 days of age, the number of external granule cells was slightly enhanced. To discriminate between the effects of the noradrenergic deafferentation and the nonspecific toxic effects of 6-OHDA, these results were compared to those obtained after an infusion of 6-OHDA + bovine serum albumin (BSA) into the locus ceruleus (which produced a selective deafferentation) and after an injection of 6-OHDA directly into the IVth ventricle (which increased the nonspecific toxic effects). Whatever the procedure used, the same reduction in the size of the vermis was observed. However, the fissuration index was not modified when BSA was added to the 6-OHDA solution. We conclude that noradrenergic deafferentation by itself was sufficient to reduce the growth of the cerebellum at certain periods of postnatal life. PMID- 2984041 TI - [Aspects of the mediator nature of the effect of delta-9-tetrahydrocannabinol on evoked potentials of the cerebral cortex]. AB - It has been shown on cats with the method of the conditional defensive reflexes and parallel recording of the bioelectric activity that the serotonin-negative and dopamine-positive substances restore the similarity of the amplitude temporary parameters of evoked potentials in the visual, associative and the second comatosencory fields of the brain cortex, disturbed by delta-9 tetrahydrocannabinol injection. It has been also demonstrated that the serotoninergic substances influence the action of delta-9-tetrahydrocannabinol both on the early and late waves of evoked potentials whereas the dopaminergic substances influence the drug action only on the late waves. PMID- 2984042 TI - The apparent non-linearity of the relationship between the rate of respiration and the protonmotive force of mitochondria can be explained by heterogeneity of mitochondrial preparations. AB - The apparent non-linear relationship between the rate of respiration and the protonmotive force in mitochondria under resting state conditions is an observation which has led to concepts such as non-chute characteristics of the proton leak through the mitochondrial membrane, or a slippage of proton pumps. We propose that this non-linearity may be a consequence of a heterogeneity of isolated mitochondria concerning the degree of coupling, since a small proportion of totally uncoupled, or loosely coupled, organelles may considerably contribute to the total respiration but not, or only slightly, to the protonmotive force. This hypothesis is supported by a fairly good fitting of computed relationships with those determined experimentally. PMID- 2984043 TI - Resonance Raman spectroscopy and enhanced photoreducibility for the 420 nm pulsed form of cytochrome oxidase. AB - Resonance Raman (RR) spectra, with 413.1 nm Kr+ laser excitation, are reported for cytochrome oxidase in resting, reduced, and 428 nm (oxygenated) forms, and for the first time, in the 420 nm (pulsed) forms [(1984) J. Biol. Chem. 259, 2073 2076]. The differences between the resting, 420 nm, and 428 nm forms' RR spectra are small. All these forms contain FeIII only, as indicated by single v4 bands at approximately 1371 cm-1, and the reoxidized forms show partial conversion from high- to intermediate- or low-spin heme a3 (intensity shift from 1575 to 1588 cm 1 for v2). The 420 nm form differs strikingly from both the 428 nm and resting forms, however, in being much more readily photoreduced by the laser illumination. This property is linked to the protein conformational change believed to be responsible for the greater accessibility to exogenous ligands of the heme a3 in the 420 nm form. PMID- 2984044 TI - Low affinity of the receptor for 1 alpha,25-dihydroxyvitamin D3 in the marmoset, a New World monkey. AB - Circulating levels of 1,25-dihydroxyvitamin D are 10-fold higher in the marmoset, a New World monkey, than in man; to assess hormone receptors, we evaluated interactions of 1,25-dihydroxyvitamin D3 with virus-transformed lymphocytes. Soluble extracts of transformed lymphocytes from humans showed hormone binding with affinity and capacity similar to that of receptors for 1,25-dihydroxyvitamin D from other human tissues. However, soluble extracts of transformed lymphocytes from the marmoset showed a strikingly lower affinity for 1,25-dihydroxyvitamin D (Kd 2.2 vs 0.27 nM in marmoset vs human) and a mildly lower binding capacity (6.9 vs 16 fmol/mg protein). A defective receptor for 1,25-dihydroxyvitamin D3 could account for resistance of target tissues to this hormone in the marmoset. PMID- 2984045 TI - Sialosyllactotetraosylceramide, a novel ganglioside antigen detected in human carcinomas by a monoclonal antibody. AB - A novel ganglioside was detected in a small cell lung carcinoma by TLC immunostaining of gangliosides with a monoclonal antibody, the C-50 MAb. Structural characterization showed this ganglioside to be IV3NeuAc-LcOse4Cer, a hitherto unknown ganglioside. This ganglioside has also been detected as a minor component in many different carcinomas using the C-50 MAb. The normally dominant CA-50 ganglioside antigen is IV3NeuAc, III4Fuc-LcOse4Cer. Based upon solid-phase binding to IV3NeuAc, III4-LcOse4Cer and IV3NeuAc-LcOse4Cer it is concluded that the C-50 MAb recognizes an epitope present in sialylated type I carbohydrate chains. PMID- 2984046 TI - 'Red pigment' from ADE-2 mutants of S. cerevisiae prevents DNA cleavage by restriction endonucleases. AB - Protection of DNA from cleavage by restriction endonucleases EcoRI, HindIII, BamHI, and Bg/II with red pigment, produced by ADE-2 mutants of Saccharomyces cerevisiae is demonstrated. Purification of yeast DNA from pigment can be achieved by chromatography on hydroxyapatite columns. PMID- 2984047 TI - Characterization of restriction-modification enzymes Cfr13 I from Citrobacter freundii RFL13. AB - This communicatiopn describes some properties of RCfr13 I and MCfr13 I, isolated from Citrobacter freundii RFL13. RCFfr13 I restriction enzyme recognizes the 5'-G GNCC sequence and cleaves, as indicated by the arrow. MCfr13 I methylase modifies the internal cytosine producing m5C (5'-GGNm5CC). RCfr13 I is sensitive not only to this type of substrate modification but also to hemimethylation in overlapping sites by MCfr10 I (internal cytosine of RCfr13 I recognition is methylated) and MHpa II (external cytosine is methylated). From these results the sensitivity of RCfr13 I to methylation by dcm methylase of E.coli in overlapping sites is deduced. PMID- 2984048 TI - Three-dimensional structure of the ribonuclease T1 X 3'-guanylic acid complex at 2.6 A resolution. AB - The mother enzyme of RNase T1 was co-crystallized with its natural product, 3' GMP at pH 4.0. The X-ray structure of this complex was refined with 2432 reflections in the 5.4-2.6 A range using a stereochemical restrained method (conventional R = 27.4%). The overall polypeptide chain folding is very similar in the secondary structure elements to the RNase T1 in the complex with 2'-GMP crystallized also at pH 4.0, but larger conformational changes occur in the loop regions. The base recognition scheme is identical in both complexes but in RNase T1 X 3'-GMP, the ribose phosphate is not seen in the electron density, probably due to static disorder. PMID- 2984049 TI - Alcohol dehydrogenase II and fructose-1,6-bisphosphatase appear to be co regulated in wild-type yeast. AB - An activity gel assay for fructose-1,6-bisphosphatase (FBP), the enzyme catalyzing the final step in gluconeogenesis in yeast, has been developed which can be used in conjunction with spectrophotometric assays to show that it is tightly co-regulated with the inducible alcohol dehydrogenase, ADHII. Both enzymes are repressed coordinately in aerobically grown yeast by the addition of high levels of glucose or ethanol, and induced on minimal medium by the addition of yeast extract. A mutant deficient in FBP segregates independently of the ADHII structural gene locus. This phenomenon is of interest because of the discovery of Ciriacy [(1979) Mol. Gen. Genet. 176, 427-431] of mutants (ccr, or carbon catabolite repression) which repress both FBP and ADHII simultaneously, along with several other enzymes. PMID- 2984050 TI - Lipid requirements for reconstitution of the delipidated beta-adrenergic receptor and the regulatory protein. AB - The role of lipids in the interaction of the beta-adrenergic receptor (R) with the regulatory protein (Gs) was investigated. Solubilized preparations of R and of Gs from turkey erythrocytes were delipidated by gel filtration. They were subsequently combined and reconstituted by the addition of various lipids. When reconstitution was carried out in the presence of soybean lipids, Gs could be fully activated via R by addition of hormone plus GTP gamma S. In contrast, purified phospholipids or a phospholipid fraction from soybean failed to produce an active system. Fractionation of soybean lipids revealed that acetone-soluble neutral lipids are essential for the reconstitution of a hormone responsive system. The acetone fraction could be replaced by specific neutral lipids such as alpha-tocopherol or cholesteryl arachidonate while a mixture of phosphatidylethanolamine, -choline and -serine satisfied the phospholipid requirement of the system. PMID- 2984051 TI - Adenylate cyclase and a fatty acid spin probe detect changes in plasma membrane lipid phase separations induced by dietary manipulation of the cholesterol:phospholipid ratio. AB - Rats fed with a cholesterol supplement to their diet exhibited an increase in their plasma membrane cholesterol phospholipid (C/P)-lipid molar ratio from 0.72 to 0.98, whereas those fed the hypocholesterolaemic drug clofibrate in their diet exhibited a decrease in this ratio to 0.62. The properties of these membranes were analysed with regard to ligand-stimulated adenylate cyclase activity and the mobility of a fatty acid spin probe which allowed lipid phase separations to be identified. Membranes with elevated C/P ratios exhibited two distinct lipid phase separations, one at around 36 degrees C that was attributed to the external half of the bilayer and one at around 22 degrees C which was attributed to the inner half of the bilayer. Membranes with lowered C/P ratios exhibited a single lipid phase separation occurring at around 21 degrees C which was attributed to the lipids of the inner half of the bilayer. These results were compared with those obtained by manipulation of C/P ratios in vitro using liposome-cholesterol exchange techniques. Dietary manipulation of the C/P ratio of plasma membranes in vivo led to alterations in the fold stimulation of adenylate cyclase by various stimulatory ligands. PMID- 2984053 TI - Sperm parameters and ejaculation before and after operative treatment of patients with germ-cell testicular cancer. AB - In testicular tumor patients, the stage of disease is most accurately determined by retroperitoneal lymph node dissection (RLND) and histologic examination of the operative specimen. The resection of possibly metastatic nodes is essential for securing a good prognosis. However, the most frequently encountered loss of ejaculatory function is a major disadvantage of radical RLND. In patients who intraoperatively proved to be free of metastases, we successfully employed a modified technique aiming at the preservation of sympathetic nerve fibers which mediate ejaculation. In 29 of 37 patients (78%), postoperative ejaculation was antegrade. It was retrograde in 5 of 37 patients (13.5%); 3 patients had no emission. These results are retrospectively compared with results of radical RLND in 87 patients of whom 64% suffered a total loss of ejaculation and only 23% had antegrade ejaculation. RLND did not affect sperm count, motility, and cell morphology. Potentia coeundi was preserved in all cases, although 27% of patients experienced diminished erectility or libido. PMID- 2984052 TI - Identification of the stable free radical tyrosine residue in ribonucleotide reductase. A sequence comparison. AB - The small subunit of ribonucleoside diphosphate reductase contains a unique tyrosine radical and a binuclear iron center. An alignment of different primary structures of the small subunit in Escherichia coli, the marine mollusc Spisula solidissima, Epstein Barr and Herpes simplex viruses shows that regions comprising residues 115-122, 204-212 and 234-241 (in E.coli numbering) are strikingly similar and are likely to be recognized as functionally important. Two of 16 tyrosine residues and 2 of 8 histidine residues are conserved. We propose that Tyr-122 is responsible for radical stabilization and that His-118 and His 241 together with Glu-115 and Asp-237 or Glu-238 are ligands of the iron center. PMID- 2984054 TI - [Effect of alpha- and beta-adrenoreceptor blockade on the secretory process in the rat pancreas]. PMID- 2984055 TI - Smoking cessation by patients with coronary artery disease. PMID- 2984056 TI - Establishment and characterization of an Epstein-Barr virus nuclear antigen negative hairy cell line. AB - Continuous cell lines have been established from spleen cells of patients with confirmed hairy cell leukemia (HCL). One cell line, HCL-Z1, lacks Epstein-Barr virus nuclear antigen (EBNA), grows attached to the substratum and retains typical features of hairy cells as revealed by transmission and scanning electron microscopy. HCL-Z1 differs morphologically from the three other EBNA-positive lymphoblastoid cell lines obtained (HCL-Z2, HCL-Z3, HCL-Z4) as well as from normal spleen cells or lymphocytes. The three lymphoblastoid cell lines derived from HCL patients show similar surface features as a line from a myeloma patient. Therefore, not all cell lines derived from HCL patients may be considered as representative of the patients leukemia cells. PMID- 2984057 TI - Glucagon-stimulated cyclic AMP production and formation of estradiol in Sertoli cell cultures from immature rats. AB - Addition of glucagon to the incubation medium of cultured Sertoli cells isolated from immature (19-day-old) rats resulted in a time- and concentration-dependent stimulation of cAMP accumulation measured both in the cells and in the medium. Maximal intracellular levels of cAMP were reached after 30 min, after which the levels decreased. In the medium cAMP levels reached a plateau after 6 h. The magnitude and kinetics of the responses were comparable to those observed with FSH in the same culture preparations. 1-Methyl-3-isobutylxanthine (MIX), a phosphodiesterase inhibitor, greatly potentiated the magnitude of the effects of glucagon and FSH. Glucagon stimulated adenylate cyclase activity in isolated membrane preparations from similar cultures, and the concentration causing half maximal stimulation (EC50) was approximately 300 ng/ml. Glucagon also stimulated aromatization in cultured Sertoli cells to the same extent as FSH. It is concluded that cultured Sertoli cells isolated from immature rats contain receptors for glucagon, coupled to adenylate cyclase, and that glucagon also stimulates aromatization of testosterone to estradiol. PMID- 2984058 TI - Characterization of the prolactin receptor in cell fractions from rat liver. AB - [125I]Prolactin (PRL) was covalently cross-linked to its binding sites in subcellular fractions of female rat livers using NHSAB and UV irradiation. Analysis by non-reducing SDS-PAGE showed that all fractions with specifically bound radioactive hormone contained a major autoradiographic band (eliminated with unlabeled PRL) of similar electrophoretic mobility consistent with a MW of 36K for the receptor. In addition, microsomal membranes were treated with a zwitterionic detergent (CHAPS), solubilizing 30-60% of the specifically bound radioactivity. SDS-PAGE analysis of [125I]PRL cross-linked to CHAPS-soluble and CHAPS-insoluble material showed an autoradiographic band with a similar MW. These results suggest that prolactin receptors in different hepatocyte membranes are similar in MW and do not appear to be linked by disulfide bonds to other membrane proteins. Some of the binding sites appear to interact with membrane constituents in ways that affect their solubility by CHAPS. PMID- 2984059 TI - Involvement of the regulatory protein (Ns) in the maturation of ACTH-sensitive adenylate cyclase of ovine fetal adrenal during late gestation. AB - The responsiveness to several stimuli of the adenylate cyclase of crude adrenal membranes from fetal and neonate lambs has been measured under different conditions. The response to Gpp(NH)p, ACTH1-24 + Gpp(NH)p and NaF was significantly lower in membranes from fetuses than from neonate lambs, whereas the response to forskolin was similar. Addition of human erythrocyte ghosts to fetal adrenal membranes enhanced the stimulation of cyclase by NaF and Gpp(NH)p, which became similar to that observed in membranes from neonates. However, such a "complementation' did not enhance the response of the enzyme to ACTH1-24. The adenylate cyclase activity displayed a biphasic response to GTP under basal conditions or when stimulated by forskolin. The magnitude of the inhibition achieved with 10(-3) M GTP was similar in fetal and neonatal adrenal membranes. These data, together with previous results (Endocrinology, 1981, 108, 2114-2119), show that the maturation of the ACTH-sensitive adenylate cyclase system of the ovine fetal adrenal gland involves an enhancement of the number of ACTH receptors and a development of the Ns subunit, whereas the catalytic site is hardly (if at all) involved in this process. PMID- 2984060 TI - Regulation of the biosynthesis of cytochromes P-450 involved in steroid hormone synthesis. AB - The actions of ACTH to regulate the synthesis of the various enzymes involved in steroid hormone biosynthesis have been studied using bovine adrenocortical cells in monolayer culture. ACTH causes an increase in the synthesis of both the mitochondrial and the microsomal forms of cytochrome P-450 involved in steroid hormone biosynthesis, as well as of the iron-sulfur protein involved in transferring electrons to the mitochondrial forms of cytochrome P-450, namely, adrenodoxin. This increased synthesis is reflective of an increase in translatability of mRNA species specific for these various proteins, and appears in each case to be mediated by cyclic AMP. Whereas the mitochondrial proteins are synthesized as precursors of higher molecular weight which are processed upon insertion into the mitochondria, the microsomal proteins are synthesized as species identical in molecular weight to the mature forms. In order to determine whether the action of ACTH to increase the rate of synthesis of these proteins is the result of an increase in the levels of specific mRNA species, cDNA clones complementary to these mRNA species are being isolated. These probes will also make it possible to characterize the genes encoding the steroidogenic enzymes, as well as to identify regulatory elements which control their transcription. PMID- 2984061 TI - Platelet-derived growth factor. PMID- 2984062 TI - Characteristics of somatotropic growth hormone binding to homologous liv plasma membranes. AB - The binding of 125I-labeled oGH, rGH and hGH to homologous liver membranes is compared to define the characteristics of somatotropic hormone-receptor interactions. The effect of pH and cations and the time-course of binding of these different GHs was similar when binding was determined in liver membranes from the homologous species. The time-course of binding was characterized by stable binding by 90-120 min of incubation at 22 degrees C with relatively high nonspecific binding that continued to increase with time. This resulted in a peak of specific binding for each hormone. Addition of excess unlabeled hormone at 30 min of the binding incubation resulted in partial displacement of the bound GH. The binding of rGH was somewhat dependent upon the presence of divalent cations (Mg2+, Ca2+) in the incubation medium. In the absence of added divalent cation, a small but appreciable amount of binding was detected at pH 7-8. With 10 mM calcium, binding increased several-fold with a definite peak of rGH binding at pH 4-5. Both oGH and hGH exhibited a broad peak of binding at pH 5-7. Monovalent cations (Na+, K+) were either ineffective in enhancing GH binding or produced a slight enhancement at higher concentrations (100 mM). Competitive displacement curves confirm the somatotropic nature of the GH-receptor interaction. Human P1 was ineffective in competing for GH binding. We conclude that 'somatotropic' growth hormone binding to its homologous liver receptor exhibits phylogenetically consistent and distinguishing characteristics of binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984063 TI - Cellular location of cytosolic triiodothyronine binding protein in primary cultures of fetal rat brain. AB - The evolution of a cytosolic triiodothyronine (T3) binding protein was studied in primary cultures of fetal rat brain. These cultures exhibited neuronal characteristics during the first week. T3 binding activity in cell supernatants increased during this period from 39 +/- 7 (mean +/- SD) to 159 +/- 24 fmoles T3/culture flask. A similar increase was observed in the soluble proteins. After day 8, neuronal death occurred and glial cells multiplied and differentiated. On day 11 an 86% drop in the binding activity was observed (24 +/- 7 fmoles T3/culture flask); the pool of soluble proteins remained stable. Scatchard analysis revealed two types of binding site in both 7- and 14-day cultured cell cytosols. Binding affinities were similar in both cytosols (KA1 approximately 1.5 X 10(9) M-1, KA2 approximately 1 X 10(8) M-1); in contrast, the number of sites was 4-fold smaller in 14-day cytosols. In subcultures mostly composed of glial cells, almost the same affinities were measured, but the numbers of both types of sites were 20 times smaller than in 7-day cells. These results show that in cell cultures from embryonic rat telencephalon, cytosolic T3 binding protein is mainly located in the neurons. PMID- 2984064 TI - Influence of follicular atresia on LH-induced cAMP and steroid synthesis by bovine thecae interna. AB - The aim of the present study was to examine the interrelationships between the luteinizing hormone (LH) receptor, the LH-induced changes in adenosine cyclic 3', 5'-monophosphate (cAMP) and steroid synthesis in theca interna tissue of large antral follicles (greater than or equal to 8 mm diameter) from oestrous cycling cows. Three distinct types of theca interna were identified (types I, II and III), all of which contained an LH receptor: type I was capable of secreting increased amounts of cAMP dehydroepiandrosterone, androstenedione and testosterone when exposed to LH; type II was capable of secreting increased amounts of cAMP and progesterone but not the androgens when exposed to LH; type III was incapable of cAMP or steroid synthesis when exposed to LH. Follicles with type I thecae contained: a full complement of granulosa cells; high intrafollicular concentrations of oestradiol; and granulosa cells with a high capacity to metabolise testosterone to oestradiol. These follicles were considered to be non-atretic structures. Follicles with types III thecae contained: fewer granulosa cells; low intrafollicular concentrations of oestradiol; and granulosa cells with a low capacity to metabolise testosterone to oestradiol. Moreover, follicles with type III thecae contained the highest concentrations of progesterone and the lowest concentrations of androstenedione and testosterone. These follicles were considered to be severely atretic structures. Follicles with type II thecae contained granulosa cell populations and progesterone, and androgen concentrations which were intermediate between those with thecae of types I and III. These follicles were considered to be at an intermediate stage of atresia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984065 TI - The functional activity of adult mouse Leydig cells in monolayer culture: effect of lipoproteins, pregnenolone and cholera-toxin. AB - Further studies were carried out on purified mouse Leydig cells to determine why they lose their hormone responsiveness after several days in monolayer culture. The effects of cholera-toxin on cyclic AMP and testosterone production were examined. It was found that cyclic AMP production could still be maximally stimulated by cholera-toxin after 4 days in culture when response to luteinizing hormone (LH) has declined. Testosterone production was, however, not maintained. Because this decline in testosterone production may have been due to the lack of a suitable substrate after several days in culture, cells were cultured initially in the presence of exogenous pregnenolone and low-density lipoproteins (LDL). Both substances were found to enhance basal and LH-stimulated testosterone production and to extend responsiveness of the cells until at least day 4, but by day 7 response was lost. Cells were then cultured in the presence of rat and human LDL and HDL and in both cases LDL was found to enhance consistently testosterone production, but HDL was much less effective. Scanning and transmission electron micrographs showed that changes in cell shape occurred during culture, but indicated that the cells were not depleted of lipid droplets by the end of culture or after LH stimulation. It is concluded that the eventual decline in testosterone synthesis is not due to lack of substrate, although the addition of exogenous substrate does extend the period of responsiveness. Nor is it due to a decrease in adenylate cyclase activity. At least part of the lesion is caused by a decrease in the enzymes required for the conversion of pregnenolone to testosterone. PMID- 2984066 TI - Characterization of immunoreactive beta-endorphin secreted from cultured human corticotropin-secreting adenomas. AB - Seven human corticotropin-secreting adenomas causing Cushing's disease or Nelson's syndrome were maintained in long-term culture. Pooled media from the individual adenomas were analyzed for the composition of their secretory products. From a radioimmunoassay (RIA) with 100% cross-reactivity for human beta endorphin (beta h-EP) and beta-lipotropin (beta h-LPH), immunoreactive beta h-EP (IR X beta h-EP) was found to be the predominant secretory product after Sephadex G-50 analysis in 4 cases (40-80% of total IR), immunoreactive beta h-LPH (IR X beta h-LPH) predominated in 1 case, and both were equipresent in 2-cases. IR X beta h-EP was further purified by high-performance liquid chromatography (HPLC) and analyzed in 4 cases with ion-exchange chromatography on SP-Sephadex C-25 and a RIA which completely cross-reacts with beta h-EP, [N alpha-Ac]-beta h-EP, beta h-EP-(1-27) and [N alpha-Ac]-beta h-EP-(1-27). In all cases, the IR X beta h-EP was the main component (40-70%); the remaining IR material was attributable partially to [N alpha-Ac]-beta h-EP or other, less defined immunoreactive material. In 3 cases, enough IR X beta h-EP material was available for HPLC and to perform a radioreceptor assay using tritiated beta h-EP as primary ligand. The displacing potency of these preparations relative to synthetic beta h-EP was related to the content of the immunoreactive component eluting in the position of synthetic beta h-EP. PMID- 2984067 TI - Morphological and functional characteristics of islets neoformed during tissue culture of fetal rat pancreas. AB - Cell suspensions prepared by collagenase digestion of the pancreas of rat fetuses (21.5 days) were cultured for 7-9 days in RPMI medium containing 10 mM glucose. Exocrine cells disappeared rapidly, whereas fibroblasts and endocrine cells proliferated. These latter were first arranged in monolayers but progressively reorganized in neoformed islets essentially composed of B-cells. Total insulin content of the culture dishes increased until day 9, and fractional insulin release was about 20% per day. After 1 week, islets incubated in glucose-free medium released less than 1% of their insulin content over 2 h. Glucose (16.7 mM) caused a slower and weaker (3-fold) stimulation than 10 mM leucine or arginine (3 5-fold). The effects of the three secretagogues were potentiated by theophylline, but only those of glucose and leucine were inhibited by diazoxide. These neoformed islets thus retain a fetal character (relatively low responsiveness to glucose), but the stimulus-specificity of the inhibition by diazoxide is the same as in adult islets. This technique may be useful for studying the mechanisms which govern the organization of pancreatic endocrine cells in islets, and which underlie their functional maturation during the perinatal period. PMID- 2984068 TI - Activity of alkylated prolactin and human growth hormone in receptor and cell assays. AB - The disulfide bonds of two lactogenic hormones, ovine prolactin (oPRL) and human growth hormone (hGH), were reduced with dithiothreitol under denaturing conditions and alkylated with iodoacetic acid. The modified hormones were assayed for their ability to bind the plasma membrane-bound receptor for lactogenic hormone found in the rabbit mammary gland. S-Carboxymethylated ovine prolactin (SCM-oPRL) with all six cysteine residues modified had a nearly 300-fold decrease in binding as compared to native oPRL in a competitive binding assay using [125I]ovine prolactin. The S-carboxymethylated human growth hormone (SCM-hGH) had all four of its cysteine residues modified. It showed only a slightly reduced ability to bind the rabbit mammary gland prolactin receptor in a competitive binding assay with [125I]ovine prolactin. The two modified hormones were assayed for their ability to stimulate proliferation of the lactogen-dependent Nb 2 lymphoma cell line. SCM-oPRL required concentrations greater than 1 X 10(5) that of native oPRL to stimulate 50% of the maximum cell growth. SCM-hGH retained a significant amount of its ability to stimulate the Nb 2 lymphoma cells. PMID- 2984069 TI - Sodium-potassium-ATPase activity in the dorsal root ganglia of rats with streptozotocin-induced diabetes. AB - Sodium-potassium-ATPase activity was measured in excised dorsal root ganglia of streptozotocin-diabetic rats, 2 months after induction of diabetes. In comparison with age-matched controls, there was a decrease in both the total and ouabain insensitive activity, indicating an overall reduction in ouabain-sensitive activity of 46%. This decrease may explain the reduced amino-acid uptake exhibited by diabetic sensory ganglia and could be relevant to the development of diabetic neuropathy. PMID- 2984070 TI - [Adrenocorticotropic effect on the activities of human kidneys]. PMID- 2984071 TI - Activation of c-myc gene transcription by Rous sarcoma virus infection. AB - Transcription of the c-myc gene in chick embryo fibroblast cells was activated 50 to 100-fold by Rous sarcoma virus (RSV) infection. More than 30-fold activation of c-myc gene transcription was also observed in RSV-transformed rat cells as compared with the control cells. The increased expression of c-myc gene in these cells was not due to gene amplification or gene rearrangement. This striking increase in expression of c-myc gene may be relevant to neoplastic transformation of RSV-infected cells. PMID- 2984072 TI - Experimental infection of rabbits with human T-cell leukemia virus type I. AB - Rabbits were successfully infected with human T-cell leukemia virus type I (HTLV I) and produced antibodies to adult T-cell leukemia-associated antigens (ATLA) on intravenous inoculation of the HTLV-producing human cord T-cell line MT-2, or autologous cell lines established by cocultivation with MT-2 cells. Lymphocytes taken from the rabbits between 4 and 14 days after the inoculation of MT-2 cells, but not lymphocytes obtained in earlier or later periods, could be immortalized in vitro and expressed ATLA and type C virus particles. However, lymphocytes harvested during an early culture period (5 to 8 days) were found to be negative for ATLA. The transformed cells had the karyotype of a normal male rabbit. Two rabbits inoculated with the autologous HTLV-producing cell lines did not allow their growth in vivo, but some peripheral blood lymphocytes could be immortalized in vitro. One of these transformed cell lines was examined for the integration of HTLV-I provirus genome. The transformed cells were found to contain the provirus genome and also to be monoclonal with respect to the integration site of the provirus genome, unlike the inoculated cells. PMID- 2984073 TI - Isoferritins from tumorous and non-tumorous human liver. AB - Human ferritins purified from normal liver (LF) and from hepatocellular carcinoma (HF) showed different biochemical and immunological characteristics. Subunit preparations from the isoferritins showed differences in isoelectric focusing patterns, although both HF and LF were found to be composed of H and L subunits with apparently the same molecular weights. It is postulated that heterogeneities between the two isoferritins are not merely due to different proportions of H and L subunits, but also result from the existence of distinct species of the subunits bearing different surface electric charges. PMID- 2984074 TI - [Sensitivity of fine-needle puncture-biopsy in the histologic diagnosis of hepatic and pancreatic malignant tumors]. AB - Aspiration-biopsies under ultrasound guidance, using a fine needle (external diameter inferior to 1 mm) have been performed in 61 malignant hepatic tumors (group I) and 25 pancreatic carcinomas (group II). Malignancy was diagnosed in 83 p. 100 cases; in group I, the sensitivity is 87 p. 100; the difference between hepatocellular carcinoma and liver metastases could be made in 88 p. 100 cases and in case of metastases, the site of the primitive carcinoma could be suggested in 77 p. 100 cases. In group II, the sensitivity is 72 p. 100. No complications were noted. PMID- 2984075 TI - [Simultaneous detection of IgM antibodies against the hepatitis A virus and the viral capsid antigen of Epstein-Barr virus in acute hepatitis]. AB - The simultaneous detection of IgM antibodies to hepatitis A virus (anti-HAV IgM) and IgM antibodies to viral capsid antigen (anti-VCA IgM) of Epstein-Barr virus (EBV) in patients with acute viral hepatitis has led us to systematically study serological markers of EBV in patients with anti-HAV IgM positive acute hepatitis and to test for anti-HAV IgM in sera of patients with acute hepatitis associated with serological evidence of current primary EBV infection. All patients studied were HBsAg negative and were not drug-addicts, nor homosexuals. In 15 consecutive patients with anti-HAV IgM positive acute hepatitis, anti-HAV IgM and anti-VCA EBV IgM antibodies were simultaneously detected in 9 cases. Of these 9 patients, antibodies to nuclear antigen were positive in 8 cases, antibodies to early antigen were positive in 7 cases and rheumatoid factor was positive in 4 cases. In 5 consecutive patients with acute hepatitis associated with serological evidence of current primary EBV infection, anti-HAV IgM was not detected. Simultaneous presence of anti-VCA EBV IgM, early antigen IgG antibodies and nuclear antigen antibodies in 7 patients with acute hepatitis associated with anti-HAV IgM suggests reactivation of EBV or reactivation of clones secreting antibodies anti-EBV in HAV infections. Furthermore, these results show that anti VCA IgM only cannot be considered to be a specific marker of early EBV infection in patients with acute hepatitis. PMID- 2984076 TI - [Presence of papilloma group viral antigen in an esophageal condyloma in man]. AB - A case of esophageal Papilloma is reported. The viral condyloma was diagnosed by standard histological methods and confirmed by immuno-fluorescence and immuno peroxidase technics against genus specific Papilloma virus antigens (i. e. capsid specific proteic antigen). These antigens were localized in the nuclei of koilocytic cells. These results confirm the association between proliferative lesions appearing on the mucosal epithelium and Papilloma viruses, which are suspected to be oncogenic in other malpighian tissues. PMID- 2984077 TI - Esophageal infections in immunosuppressed patients after marrow transplantation. AB - We reviewed all cases of infectious esophagitis diagnosed after marrow transplantation during a 2-yr period. Twenty-one infections were found during 46 fiberoptic endoscopic examinations; most infections were viral, with equal numbers due to cytomegalovirus and herpes simplex virus. These infections presented with nonspecific esophageal symptoms weeks to months after transplantation. Diagnoses depended on histology and viral cultures. During the same time span, autopsies revealed 25 cases of infective esophagitis among 59 patients; most were again viral, with equal numbers of herpes simplex virus and cytomegalovirus infections. Mixed infections due to viral, fungal, and bacterial pathogens were found in both endoscopy and autopsy series. The prevalence of esophageal infection in marrow transplant patients with esophageal symptoms is high, but the organisms could not be predicted on the basis of symptoms, oropharyngeal surveillance cultures, or esophageal x-ray. Accurate diagnosis requires that endoscopic biopsy specimens and brushings be examined by both histologic and virologic methods. PMID- 2984078 TI - Role of oxygen-derived free radicals in hemorrhagic shock-induced gastric lesions in the rat. AB - This study was designed to determine whether oxygen-derived free radicals play a role in the pathogenesis of gastric lesions produced by hypotensive ischemia in the rat. To achieve this goal, allopurinol, an inhibitor of xanthine oxidase (the enzyme responsible for the formation of superoxide radicals); superoxide dismutase, a scavenger of superoxide radicals (O2-); and dimethyl sulfoxide, a scavenger of hydroxyl radicals (OH) were used. In the anesthetized rat, HCl (0.1 N) was instilled into the pylorus-ligated stomach, and the rat was bled to reduce the blood pressure to less than 30 mmHg. The blood pressure was maintained at less than 30 mmHg for 20 min and then the shed blood was retransfused. Twenty minutes after the retransfusion the rat was killed, the stomach was removed, and the area of gastric mucosal lesions was measured. Both allopurinol and superoxide dismutase, but not dimethyl sulfoxide, significantly protected against hemorrhagic shock-induced gastric lesions. These findings suggest that oxygen derived free radicals, particularly O2-, play an important role in the formation of gastric lesions produced by ischemia plus HCl. PMID- 2984079 TI - Effects of epidermal growth factor on acid secretion from guinea pig gastric mucosa: in vitro analysis. AB - Epidermal growth factor (EGF) has been tested on guinea pig gastric mucosa mounted in Ussing chambers to investigate the suitability of using in vitro methods for examining EGF's effects on acid secretion. Epidermal growth factor reduced the rate of histamine-induced acid secretion to near basal levels when applied to the serosal gastric surface at nanomolar concentrations. Inhibitory effects were evident 10-15 min after EGF treatment and were maximal by 40 min. Cyclic adenosine monophosphate-induced secretion was also reduced by EGF, although the effect occurred more slowly than in histamine-treated tissues. Epidermal growth factor increased transmucosal resistance in histamine-treated, but not cyclic adenosine monophosphate-treated mucosa; potential difference was unaffected. Nerve growth factor had no effect when tested in the in vitro system. The EGF binding protein was found to enhance slightly the inhibitory activity of EGF on acid secretion. When applied to the luminal (mucosal) gastric surface, EGF inhibited secretion marginally but only at micromolar concentrations. These results indicate that EGF acts directly upon cells within the gastric mucosa, and is most effective when applied to the serosal gastric surface. They further suggest that in vitro preparations of intact gastric mucosa can be used for analyzing the inhibitory effects of EGF on gastric acid secretion. PMID- 2984080 TI - Caerulein-induced acute necrotizing pancreatitis in mice: protective effects of proglumide, benzotript, and secretin. AB - The onset, course, and regression of the biochemical and structural alterations associated with pancreatitis induced by various doses of caerulein were studied in the mouse. In addition, the protective effect of secretin was compared with that of the cholecystokinin-receptor antagonists proglumide and benzotript. Subcutaneous or intraperitoneal injections of caerulein induced increases in serum amylase concentration and pancreatic weight and histologic evidence of acute pancreatitis, all effects being dose-related. Cytoplasmic vacuoles were the earliest histologic alterations. As the pancreatitis progressed these vacuoles increased to an enormous size. Interstitial inflammation and acinar cell necrosis were prominent after 6 h, reached a maximum after 12 h, and mostly disappeared after 4 days. During the course of pancreatitis approximately 40% of the acinar cells showed signs of severe degeneration or necrosis at the most effective doses of caerulein. Electron microscopy showed both intact and degenerating granules inside the vacuoles. Signs of basolateral exocytosis of zymogen granules were not observed. During the regression of pancreatitis, focal atrophy was a remarkable histologic finding. Repetitive initiation of pancreatitis (six courses of caerulein injections over 5 wk) produced marked focal atrophy and early fibrosis. High doses of proglumide or benzotript markedly ameliorated both the biochemical and structural alterations induced by caerulein. Secretin, even at very high doses, had only minor protective effects. This study presents a model of acute necrotizing pancreatitis in which the severity of the induced pancreatitis ranges dose-dependently from mild interstitial inflammation to severe necrosis. The ultrastructural alterations described herein support the hypothesis that the trigger mechanism of acute pancreatitis appears to be a primary intracellular event rather than an interstitial event that secondarily damages the acinar cells. PMID- 2984081 TI - Myeloperoxidase and intestinal inflammation. PMID- 2984082 TI - [Grading of epithelial ovarian cancers]. AB - Tissue slides of 230 epithelial ovarian cancers were examined by two different pathologic teams to determine the following details: histological pattern, grade of polymorphy, mitotic index, density of chromatin, presence of nuclei and macronuclei, lymphocytic infiltration, psammoma bodies, and necrosis. For statistical evaluations the data were presented together with stage, age and survival times in form of an information vector for each of the 230 patients and analysed using hazard-models (Cox 1972, Carter 1983) and contingency tables. RESULTS: The agreement between the 2 pathologic teams was good. There were many interrelated histological concomitants, such as polymorphy and chromatin density, and as nuclei and macronuclei. Necrosis was relatively independent. Significant effects on prognosis could be demonstrated for polymorphy, chromatin density, mitotic index, histologic pattern, (macro)nuclei. Polymorphy was the best histological prognostic indicator and, together with necrosis, produced the best prognostic grading system. PMID- 2984083 TI - [Thermobiologic evaluation of benign and malignant breast diseases]. AB - This paper is a review of the recent technical and clinical advances in the area of breast thermography. Emphasis is placed upon thermal imaging using liquid crystal films, and computer-assisted analysis of breast thermograms. New data are presented concerning the value of thermography for the identification of women at high risk of developing breast cancer, the early detection of mammary carcinomas, and the detection of cancer in fibrocystic breasts. PMID- 2984084 TI - [Comparative analysis of the protein composition of semi-finished products of human albumin treated with silochrome sorbent]. PMID- 2984085 TI - Chlorpromazine and trifluoperazine potentiate the action of adenosine on rat cerebral cortical neurons. AB - The effect of chlorpromazine and trifluoperazine on adenosine-evoked depressions of the spontaneous firing of rat cerebral cortical neurons has been studied. When applied by iontophoresis from multiple barrelled micropipettes, in amounts that did not affect neuronal firing rates, both substances enhanced the depressant actions of adenosine. Trifluoperazine did not enhance the depressant actions of the uptake resistant analog, adenosine 5'-N-ethylcarboxamide. Intravenously administered chlorpromazine and trifluoperazine (1 mg/kg) enhanced the depressant actions of iontophoretically applied adenosine. Caffeine antagonized the depressant effect on the firing of cortical neurons observed with larger applications of trifluoperazine. These findings support the conclusion, drawn from previous studies on the effects of phenothiazines on the uptake of adenosine by brain synaptosomes, that inhibition of adenosine uptake and potentiation of the actions of endogenously released adenosine may be a significant factor in the generation of the central actions of the phenothiazines. PMID- 2984086 TI - Identification of potential GABA-mimetics by their actions on brain GABA recognition sites. AB - In an attempt to identify potential anticonvulsant compounds, 18 structural analogues of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) were tested for their ability to inhibit GABA receptor binding, sodium-dependent GABA binding and GABA aminotransferase activity in synaptic membranes from mouse brain. Nine inhibitors of receptor binding were found. The most potent was N (thiocarbamoyl)glycine (Ki = 18 microM). However, this compound had no real effect on Na+ -dependent GABA binding nor on the activity of GABA aminotransferase. In addition, it was unable to enhance the binding of [3H]flunitrazepam as GABA agonists usually do. This could indicate that this inhibitor is, rather, a GABA receptor antagonist. Even though no particularly potent inhibitors of any of the GABA recognition sites were found, this technique nevertheless demonstrates how simple in vitro assays can be used to find drugs exhibiting potential GABA-mimetic activity. PMID- 2984087 TI - Protective effects of "cerebroactive drugs" in a model of acute hypoxia. AB - A systematic study of the hypobaric hypoxia method was carried out, using a wide range of vasodilators and metabolic modifiers. In general cerebral metabolic modifiers have a more effective antihypoxic action than cerebral vasodilators. After a discussion, the conclusion is that hypobaric hypoxia is useful as an initial screening procedure. PMID- 2984088 TI - Separation and partial characterization by high-performance liquid chromatography and radioimmunoassay of different forms of melanocyte-stimulating hormone from fish (Cyprinidae) neurointermediate lobes. AB - From neurointermediate lobe (NIL) extracts of two species of Cyprinidae, Carassius auratus and Cyprinus carpio, several peptides were separated by high performance liquid chromatography (HPLC) on a C18 muBondapak column eluted with a methanol/acetic acid/triethylamine mixture. Monitoring all fractions by radioimmunoassay (RIA) with an antibody against melanocyte-stimulating hormone (MSH) C terminal gave positive reactions for fractions 7, 11-12, 15-16, 23-24, and 25-27. For further characterization, the elution positions of these peaks were compared to those of known synthetic reference substances. Peak 7 elutes in the same position as oxidized alpha MSH, whereas peak 15-16 matches the elution position of des-acetyl alpha MSH and 23-24 that of alpha MSH. The product from peak 26-27 has several characteristics of the diacetylated form of alpha MSH: its immunoreactivity in RIA, its sensitivity to weak bases and to HCl and its mass spectrum which is identical with that of mammalian diacetyl alpha MSH. In both species, the diacetylated form is predominant in the intracellular pool. This study establishes the coexistence of three different forms of alpha MSH, a des acetylated, monoacetylated, and diacetylated in the cyprinid NIL extracts. PMID- 2984089 TI - RIA determination and immunofluorescence localization of cyclic nucleotides in rainbow trout (Salmo gairdneri) testes. AB - Using histological criteria, testicular development was divided into six stages (I-Va). The testicular amounts of 3',5'-cyclic adenosine- and guanosine monophosphate (cAMP, cGMP) were determined radioimmunologically, and their testicular distribution patterns were monitored by indirect immunofluorescence microscopy. During slow testicular growth (stages I-III), the nucleotide concentrations were high (0.62-1.2 pmol cAMP/mg, 0.17-0.24 pmol cGMP/mg). With the appearance of spermatozoa in stage IV, they fell to low levels which were maintained with some fluctuations until the end of the cycle (0.05-0.1 pmol cAMP/mg, 0.016-0.05 pmol cGMP/mg). The cAMP antiserum intensely stained spermatogonia, a portion of the spermatocytes, and spermatids. Spermatozoa showed almost no staining. Fluorescence labeling of somatic cells was observed in immature testes and during spermiation. Except for staining all spermatocytes, the same pattern was observed using the cGMP antiserum. PMID- 2984090 TI - Simian AIDS: evidence for a retrovirus etiology. PMID- 2984091 TI - Immunologic aspects of AIDS. PMID- 2984092 TI - Cytomegalovirus infection in homosexual men--relationship to AIDS. PMID- 2984093 TI - Gastrointestinal cryptosporidiosis and cytomegalovirus enterocolitis. PMID- 2984094 TI - [Air flow thermodestruction of chemicals produced on the basis of aromatic polyamines]. PMID- 2984095 TI - [Pathogenesis of pulmonary hypertension in pulmonary embolism]. PMID- 2984096 TI - Neuroendocrine islet cell tumour producing gastrin and ACTH in a patient with calcifying chronic pancreatitis. AB - A patient with a calcifying chronic pancreatitis was found to have a neuroendocrine islet cell tumour (a previously unreported association). The tumour secreted both gastrin and ACTH leading to clinical manifestations of both the Zollinger-Ellison syndrome and Cushing's syndrome. PMID- 2984097 TI - Immunopathogenesis of experimental Coxsackievirus induced myocarditis: role of autoimmunity. AB - The pathogenesis of cardiac injury in clinical myocarditis is unknown. Despite the association of the disease with recent viral infections, it is now assumed that immune rather than viral mechanisms are primarily responsible for myocyte destruction. Nonetheless, immunosuppressive therapy has not been universally effective in limiting myocardial damage. To better understand the mechanisms by which viral infections of the heart induce myocarditis, it has been necessary to resort to a murine model of the disease. When inbred Balb/c mice are infected with a cardiotropic variant of Coxsackievirus, group B, type 3 (CVB 3), the animals develop extensive interstitial and focal inflammatory cell infiltration of the heart similar to the lesions in humans. As in humans, a number of factors influence the severity of the disease. Males develop severe myocarditis while virgin females are generally protected. Female resistance does not persist during pregnancy, however, when resultant myocarditis is frequently worse than that observed in males. The susceptibility of males and pregnant females results from the influence of testosterone and progesterone on the immune response. Susceptible animals generate autoimmune cytolytic T lymphocytes which recognize normal myocyte cell surface antigens and are responsible for most of the cardiac damage in experimental myocarditis. Virgin females do not develop significant myocarditis apparently because the estrogens enhance suppressor cells which prevent the autoimmune T cell generation. Humoral (antibody-mediated) immunity to the heart antigens is also present, but apparently has no role in the pathogenesis of the disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984098 TI - Clinical findings in acute viral myocarditis. With special attention to experimental and immunological evidence. AB - According to analysis of 40,000 autopsies, the incidence of myocarditis is approximately 3.5%, the diagnosis of which had not been established in the majority prior to death. The clinical characteristics of myocarditis encompass a wide spectrum from an asymptomatic course to sudden cardiac death. Symptoms and findings of congestive heart failure or an inflammatory virus disease may be present. There is usually evidence of nonspecific clinical, radiologic, electrocardiographic, echocardiographic and blood chemistry abnormalities. In the presence of myocarditis, gallium scintigrams are generally pathologic. Virologic and serologic studies may have an important, albeit not decisive, role in establishing the diagnosis. In the acute phase, endomyocardial biopsy can document inflammatory changes. For the most part, treatment is symptomatic. Ventricular arrhythmias and high-grade conduction disturbances prerequisite vigilant monitoring as well as adequate medical and, if indicated, electrical therapy. In addition to conventional treatment, steroids, more recently, in combination with azathioprine, have been employed in patients with subacute and chronic myocarditis. The course of myocarditis is frequently benign although, in some cases, congestive heart failure may develop. Laboratory studies in DBA/2 mice with congestive heart failure subsequent to surviving myocarditis have shown myocardial fibrosis and hypertrophy without mononuclear cell infiltrates indicative of transition to dilated cardiomyopathy. PMID- 2984099 TI - Ovine corticotropin-releasing factor administration in normal men. Pituitary and adrenal responses in the morning and evening. AB - We administered ovine corticotropin-releasing factor (CRF) as a bolus intravenous injection (1 microgram/kg) at 09.00 and at 20.00 to assess the influence of circadian changes in the hypothalamic-pituitary-adrenal axis on the response to CRF. The increase in plasma ACTH levels after CRF was only slightly lower in the morning than in the evening. The plasma cortisol response to ACTH, however, was significantly greater in the evening than in the morning (p less than 0.005). At both times of day CRF administration had no effect on plasma concentrations of GH, PRL, LH, AVP, insulin, PRA or glucose. No effects were observed on the hematopoietic system, kidneys or liver. In addition, CRF had no effect on heart rate, blood pressure or respiratory rate at the dose employed. Approximately 10% of the subjects complained of a transient upper body and facial hot flush. These observations indicate that the magnitude of the plasma cortisol rise after CRF depends on the time of administration. PMID- 2984100 TI - Nifedipine does not impair the hormonal responses to graded exercise in healthy subjects. AB - The aim of this study was to assess whether the potent calcium antagonist nifedipine was capable of modifying the hormonal response to graded exercise in 7 healthy young men. After fasting overnight, each subject came to the laboratory on 2 consecutive mornings. On one day he was given 10 mg of nifedipine sublingually and on the other an identical placebo capsule; the order was randomised in a double-blind fashion over the 2 days. Thereafter each subject performed 2 successive short treadmill runs, equivalent to 60 and 100%, respectively, of maximal aerobic power. While significantly blunting the rise in mean systolic blood pressure and inducing a greater fall in diastolic blood pressure during and after exercise compared with the placebo, nifedipine did not impair the brisk response to pituitary-adrenal hormones (ACTH, cortisol and total catecholamines). Nifedipine also did not modify the effects of short-term exercise in raising mean plasma glucose levels, stimulating pancreatic glucagon secretion and producing a delayed increase in plasma insulin concentrations. Nor did the drug blunt the significant rise of growth hormone and prolactin levels occurring during and after the treadmill run. It was concluded that, apart from inducing significant changes in blood pressure, a single dose of nifedipine does not appear to suppress the counterregulatory hormonal responses to short-term physical activity in healthy men. PMID- 2984101 TI - Androgen and estrogen response to adrenal and gonadal stimulation in idiopathic hemochromatosis: evidence for decreased estrogen formation. AB - Gonadal function in idiopathic hemochromatosis (IHC) was evaluated by comparing clinical features and levels of sex hormones in 10 male patients with IHC (cirrhosis, 4; fibrosis, 6), 6 male patients with alcoholic cirrhosis (AC) and 10 healthy, age-matched controls. Impotence was present in 9 IHC and all AC patients and was associated with decreased plasma testosterone levels. However, gynecomastia, a feature in all patients with AC, was not present in IHC, and plasma sex hormone binding globulin was normal. Patients with IHC showed significantly lower basal estradiol levels (17.7 +/- 6.3 pg per ml) than did controls (28.5 +/- 8.5 pg per ml), and low LH levels (p less than 0.01), which were insufficiently stimulated by luteinizing hormone releasing hormone (n = 8) as well as a decrease in prolactin concentration (2.9 +/- 1.4 vs. 5.9 +/- 1.9 ng per ml in the controls) suggesting pituitary failure. Synthesizing capacity of sex hormones was determined by adrenocorticotropic hormone and human chorionic gonadotropin administration. Basal and stimulated levels of androstenedione and cortisol indicated normal function of the adrenals in IHC. However after adrenocorticotropic hormone, estrone levels increased to only 16.2 +/- 8.4 pg per ml (controls, 27.3 +/- 4.7 pg per ml; p less than 0.01). Increments of estrone (12.5 +/- 9.2 pg per ml) and estradiol (17.9 +/- 11.6 pg per ml) were also lower in IHC following human chorionic gonadotropin administration than in controls (26.0 +/- 7.2 and 37.5 +/- 11.4 pg per ml, respectively). In contrast, plasma human chorionic gonadotropin raised testosterone levels 3.3-fold in IHC and 2.2 fold in controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984102 TI - Storage of alpha-1-antitrypsin in intrahepatic bile duct cells in alpha-1 antitrypsin deficiency (Pi Z phenotype). AB - Storage of alpha-1-antitrypsin (AAT) has been found in a small number of bile duct cells in liver tissue specimens from patients with Pi MZ, Pi SZ and Pi ZZ phenotypes. The storage appeared in the form of intracellular AAT immunoreactive inclusions. On EM investigation, AAT-like material was detected within cisternae of the RER and SER. Such AAT inclusions were found in proliferating bile ductules in conditions such as cirrhosis, focal nodular hyperplasia and extrahepatic obstruction. They were also observed in normal biliary structures at the level of the canals of Hering, bile ductules and interlobular ducts in 13 out of 47 cases. These findings are interpreted as indicating that the intrahepatic bile duct cells are a further source of AAT, and that in case of defective export of AAT from the cell, as is the case for the Z protein, the protein accumulates not only in hepatocytes but in biliary cells as well. PMID- 2984103 TI - Oxytocin improves male copulatory performance in rats. AB - The role of oxytocin in male copulatory performance was reexamined in rats. Adult male rats were trained seven times for copulatory behavior, at weekly intervals. Oxytocin, either intraperitoneally injected (200 ng/rat) or intracerebroventricularly infused (1 ng/rat in 4 microliter saline) 60 and 5 min, respectively, before the eighth test, significantly shortened both the ejaculation latency and the postejaculatory interval. The intracerebroventricular infusion of saline alone (4 microliter/rat) had no effect at all. PMID- 2984104 TI - An estimate of unique DNA sequence heterozygosity in the human genome. AB - Fifteen different restriction fragment length polymorphisms (RFLPs) were detected in the human genome using 19 cloned DNA segments, derived from flow-sorted metaphase chromosomes or total genomic DNA, as hybridization probes. Since these clones were selected at random with respect to their coding potential, their analysis permitted an unbiased estimate of single-copy DNA sequence heterozygosity in the human genome. Since our estimate (h = 0.0037) is an order of magnitude higher than previous estimates derived from protein data, most of the polymorphic variation present in the genome must occur in non-coding sequences. In addition, it was confirmed that enzymes containing the dinucleotide CpG in their recognition sequence detect more polymorphic variation than those that do not contain CpG. PMID- 2984105 TI - Linkage studies between polymorphic markers on chromosome 4 and cystic fibrosis. AB - It has been suggested that a protein factor causing ciliary dyskinesis is a marker for the basic defect causing cystic fibrosis (CF), and that the structural gene for this protein may be (amongst others) on human chromosome 4. We have isolated two DNA sequences mapping to chromosome 4 which show restriction fragment length polymorphisms (RFLPs), and have followed their segregation in families in which cystic fibrosis occurs. Eleven families with a total of 30 children with CF and ten unaffected sibs were studied. We have also followed the inheritance of RFLPs revealed by two probes mapping to chromosome 4 and obtained from another laboratory, polymorphisms revealed by cloned coding sequences for albumin and fibrinogen, and the inheritance of the MNS blood group. Although the level of albumin is altered in children with CF, the gene does not segregate with CF, and therefore albumin can be excluded as the site of the basic defect. Tight linkage with CF was not found with any of the seven markers investigated, and therefore, assuming that the markers (excepting MNS and fibrinogen) are unlinked to one another, approximately half of the total genetic length of chromosome 4 may be excluded. PMID- 2984106 TI - A study of restriction fragment length polymorphisms at the human alpha-1 antitrypsin locus. AB - A cloned cDNA for alpha-1-antitrypsin (alpha-1-AT) was selected from a human liver cDNA library. The identity of the clone was established by hybrid-selected translation and partial DNA sequencing. The cDNA was used as a probe to search for restriction site polymorphisms (RSPs) near the alpha-1-AT gene. Only two RSPs were found using 29 different restriction enzymes. Each of these polymorphisms resulted from the loss of a restriction site, one for EcoRI and the other for Taq I. The frequency of polymorphic restriction was calculated to be 1.1% to 2.6% of all sites tested, a figure lower than the 9.3% value observed for 12 RSPs in the human beta-globin gene cluster. Since the corresponding figure for detectable polymorphisms at the alpha-1-AT locus at the protein level is 12%, restriction enzymes are comparatively inefficient in detecting genetic variability. The basis of this inefficiency was studied by computing the nucleotide diversity from the RSP data. On the average, one in 500 to 1000 bases is polymorphic around the alpha-1-At locus. This value is comparable to that which we have calculated for the human beta-globin gene cluster and the human growth hormone gene cluster (both one in 500). These data demonstrate the limited usefulness of linked RSPs for genetic linkage studies at the alpha-1-AT locus. PMID- 2984107 TI - DNA polymorphisms flanking the apo A-1 and insulin genes and type III hyperlipidaemia. AB - Two alleles identified by DNA restriction fragment length polymorphisms around the apo A-1/C-III and insulin genes have been shown to be associated with Type IV and V hyperlipidaemia. We have genotyped 19 patients with Type III hyperlipidaemia to establish whether this association is also found in the disorder. Our data show that these associations are not responsible for the majority of cases of Type III hyperlipidaemia, but cannot exclude the possibility that a small proportion (less than 50%) of cases of Type III are caused by interaction between these alleles and the apolipoprotein E2 phenotype. PMID- 2984108 TI - Anaerobic bacteria and herpes simplex virus in genital ulceration. AB - Of 91 patients with genital ulceration, herpes simplex virus was isolated from 52 (57%) and Haemophilus ducreyi from 12 (13%); none had syphilis. The difference in incidence of other aerobes in patients and controls was not significant. Anaerobes, predominantly Bacteroides spp, were isolated from a large proportion (77%) of men and women patients with ulcers but from few control men. The most common anaerobic species were B asaccharolyticus and B ureolyticus, with fewer isolates of the melaninogenicus/oralis group. The bacterial flora of herpetic and non-herpetic ulcers were similar, but Candida albicans was isolated significantly more often from non-herpetic ulcers. Anaerobic bacteria may contribute to the pathogenesis of genital ulcers. PMID- 2984109 TI - [Detection of macrophage activity in the blood and synovia of chronic polyarthritis patients using an activated oxygen microassay]. AB - A microassay was developed for the estimation of macrophage (M phi)-activation by released activated oxygen. From peripheral blood or synovia M phi are isolated. O 2 which is released by activated M phi is photometrically detected by cytochrome c reduction. Differences in O-2-production do not exist between monocytes of rheumatoid arthritis patients and controls. In contrast synovial M phi from rheumatoid arthritis patients show increased levels of O-2-production. PMID- 2984110 TI - Activation and proliferation signals in mouse B cells. V.A. comparison of the effects of intact (IgG) and F (ab')2 anti-mu or anti-delta antibodies. AB - This study compares the effects of soluble intact (IgG) and F(ab')2 rabbit anti mu and anti-delta antibodies on mouse B cells. The results show that while the F(ab')2 antibodies to both isotypes induce polyclonal B cell proliferation, the IgG antibodies are not mitogenic, but rather inhibit DNA synthesis induced by the homologous F(ab')2 fragments. Furthermore, intact anti-mu antibodies inhibit mitogenesis induced by F(ab')2 anti-delta, and vice versa. However, the intact antibodies to both isotypes promote early changes characteristic of B-cell activation, namely, increased Ia antigen expression, and priming for a facilitated proliferative response to F(ab')2 anti-Ig. In addition, F(ab')2 anti mu and anti-delta both induce the breakdown of phosphatidylinositol phospholipids in B cells, an early consequence of Ig receptor cross-linking which may be involved in the induction of cell growth. These results therefore indicate that stimulating mature B cells via IgM or IgD receptors produces indistinguishable early effects, and that cross-linking Fc and surface Ig receptors by intact anti Ig generates a dominant inhibitory signal, regardless of which isotype is involved. PMID- 2984111 TI - Orientation of the loci encoding RT1.B polypeptides in the major histocompatibility complex of the rat. AB - The major histocompatibility complex of the rat (RT1) contains genes that code for two class II histocompatibility antigens. The r12 rat strain (WRC) was derived from a mating which revealed a genetic recombination that defines the two loci, RT1.B and RT1.D, that code for the class II antigens. The RT1.B and RT1.D antigens of the RT1 complex are protein dimers consisting of an alpha and beta glycoprotein chain and are homologous to I-A and I-E genes, respectively, in the H-2 complex of the mouse. We have performed Southern blot analysis on liver DNA from the r12 and parental strains to examine the precise location of the recombinant event. After digestion with restriction enzymes, the DNA was separated on agarose gels, blotted onto nitrocellulose, and hybridized with mouse H-2 cDNA probes specific for I-A alpha and beta genes. The pattern of restriction fragment polymorphisms demonstrated that the site of recombination is between the RT1.B alpha and the RT1.B beta genes. As a result of these findings, we suggest that the sequence of genes within the RT1 complex consists of RT1.A ... RT1.B beta ... RT1.B alpha ... RT1.D (alpha, beta) ... Other class I genes, possibly corresponding to mouse Qa and Tla-like genes, were also apparent in these experiments. PMID- 2984113 TI - Restriction fragment length polymorphism of the major histocompatibility complex of the pig. AB - Human HLA cDNA probes were used to analyze the restriction fragment length polymorphism (RFLP) of the SLA major histocompatibility complex in swine. Cellular genomic DNA from 19 SLA homozygous pigs representing 13 different haplotypes was digested with restriction endonucleases Eco R1, Hind III, or Bam H1, separated by electrophoresis, and transferred onto diazobenzyloxymethyl paper by the Southern blot technique. The blots were probed with 32P-labeled class I or beta-DR class II cDNA. Depending on the haplotypes and the endonucleases used, seven to ten restriction fragments hybridized with the class I probe, and five to seven with the beta-DR probe. Their sizes ranged from 3.4 to 22 kilobase-pairs. Few bands were common to all 13 haplotypes. With all but one haplotype, identical autoradiogram patterns were obtained from unrelated, but phenotypically SLA identical pigs, suggesting that most of the RFLP revealed were controlled by the SLA region. Further polymorphism was found in a group of seven unrelated pigs which typed serologically as SLA A15 C1 B18 homozygotes but could be divided into two subgroups, with five animals in one subgroup and two in the other, when the genomic DNA was hybridized with the class I probe. When the class II beta-DR probe was tested on the same seven pigs, another subdivision was seen, and this correlated with MLR data. These results demonstrate that HLA class I and class II probes can be used to identify certain well-established SLA haplotypes and to identify subclasses within at least one SLA haplotype. PMID- 2984112 TI - Localization of C4 genes within the HLA complex by molecular genotyping. AB - Segregation of the complement component, C4, was analyzed in six families that each included an individual who inherited an HLA haplotype where a crossover event had occurred in the region between HLA-B and HLA-DR. Two cDNA clones corresponding to the C4 gene were utilized as probes in Southern blot analysis of DNA from members of each family. Restriction fragment length polymorphisms (RFLP) were observed and were assigned to haplotypes. In one family RFLP, hybridizing with the C4 probes, segregrated with HLA-B, and in four families RFLP segregated with HLA-DR; one family was not informative in this respect. These analyses have made it possible to localize the genes for C4 between HLA-B and HLA-DR by molecular genotyping and to characterize three different genomic configurations of C4 genes by limited restriction mapping. PMID- 2984114 TI - A 3.5 kilobase Taq I restriction fragment of genomic DNA segregates with HLA-B27. PMID- 2984115 TI - Blood pressure and hormonal responses to aerobic exercise. AB - Twelve patients with essential hypertension (WHO stages I-II) were subjected to mild aerobic exercise for 10 to 20 weeks. The time course of changes in the resting blood pressure and multiple hormonal responses (plasma catecholamines, prostaglandin E, renin-angiotensin system, kallikrein-bradykinin system) were monitored. Depressor response of both systolic and diastolic pressures was seen, and after 5 weeks of exercise blood pressure stabilized at a significantly lower level. Adjustment of work load in response to increased physical fitness at the 10th week produced further reduction of blood pressure especially in diastole. After exercise therapy we found significant reductions in plasma catecholamine levels, and increases in levels of plasma prostaglandin E and the urinary excretion of sodium. A reduction in systolic/diastolic (mean) pressures by more than 20/10 (13) mm Hg was seen in 50% of patients after 10 weeks and in 78% after 20 weeks of exercise. Those who achieved effective blood pressure fall after 10 weeks of training (n = 6) were compared with the rest (n = 6). This analysis revealed that the initial value of plasma renin activity of the former was significantly lower than that of the latter. Significant negative correlations (r = -0.78) also were observed between the blood pressure reduction and corresponding initial value of plasma renin activity. These results indicate that exercise therapy is a potent nonpharmacological tool for the treatment of essential hypertension, especially of the low renin type. Both diminished sympathoadrenergic activity and enhancement of prostaglandin mechanisms might be responsible for the falls in arterial pressure. PMID- 2984116 TI - Converting-enzyme inhibitor administration lowers urinary free 19-nor deoxycorticosterone levels. AB - 19-Nor-deoxycorticosterone (19-nor-DOC) is a human mineralocorticoid. The regulation of its secretion is poorly understood, as renin angiotensin II (ANG II) stimulation has minimal effects on 19-nor-DOC. This study sought to determine if ANG II inhibition would decrease 19-nor-DOC production. Six normal subjects on fixed electrolyte intake were admitted to a metabolic unit. After a 5-day control period to establish electrolyte balance, enalapril, p.o., 10 mg/day, was administered for 28 days. This treatment resulted in ANG II inhibition, which was reflected by a rise in plasma renin activity, a blunting of the postural plasma aldosterone increment, and a decrease in aldosterone secretion rate (ASR). Levels of urinary free (UF) 19-nor-DOC progressively decreased from 294 +/- 108 ng/day on Day 0 to 164 +/- 70 on Day 3, 141 +/- 62 on Day 7, 101 +/- 38 on Day 14, 68 +/ 18 on Day 21, and 106 +/- 31 on Day 28. The decrease in 19-nor-DOC levels was synchronous with the fall in ASR (R = 0.94, n = 5, p less than 0.005), but it was of greater magnitude (71% decrease in 19-nor-DOC levels versus 41% decrease in ASR). In addition, the decrease in 19-nor-DOC levels correlated with a fall in urinary potassium and an increase in both urinary sodium and chloride (R = 0.68, 0.79, -0.87 respectively; n = 6, p less than 0.05). The fall in ASR, on the other hand, was not significantly correlated with the changes in these urinary electrolyte levels (R = 0.65, 0.64, 0.57 respectively; n = 5).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984117 TI - Partial deficiency of adrenal 11-hydroxylase. A possible cause of primary hypertension. AB - Results of supraphysiological adrenocorticotropic hormone (ACTH) stimulation of biosynthetic pathways of adrenal zona fasciculata indicate that a deficiency of 11-hydroxylase exists in patients with essential hypertension. The deficiency is suggested by the much greater stimulus of synthesis of deoxycorticosterone (DOC) and deoxycortisol in hypertensive subjects than in controls (p less than 0.001). No significant difference in the synthesis of cortisol, corticosterone, progesterone, 17-hydroxyprogesterone (17-OHP), and delta-4-androstenedione (D4) was observed between the two groups. The ratios for synthesis of DOC and corticosterone and for deoxycortisol and cortisol found in hypertensive patients were significantly higher than those found in controls (p less than 0.001); no significant difference was observed in the synthesis of 17-OHP and progesterone. The synthesis of DOC and deoxycortisol was not significantly correlated with either blood pressure or plasma renin activity. Plasma renin activity was significantly lower in hypertensive subjects than in normotensive subjects (p less than 0.0001), while no difference was found in aldosterone secretion between the two groups. The 11-hydroxylase deficiency in the adrenal zona fasciculata may be one of the genetic factors causing hypertension together with environmental factors (particularly salt intake and work-related stress). The investigation performed in our study may be useful for the evaluation of adrenal zona fasciculata enzymatic activities during the study of hypertensive patients. PMID- 2984118 TI - An improved fluorometric assay of rat serum and plasma converting enzyme. AB - The most sensitive nonradiometric routine assay for angiotensin-converting enzyme (ACE) activity uses fluorometry to detect His-Leu released from Hip-His-Leu. Our results indicate that, in contrast to human serum, rat serum and plasma contain large and variable amounts of dipeptidase activity that lead to a subestimation of the ACE activity measured in 0.1 M potassium phosphate buffer, pH 8.3, containing 0.3 M NaCl, the most commonly used assay for human serum and tissue ACE. We describe and validate an assay for 1 to 10 microL rat and human serum or plasma using 5 mM Hip-His-Leu in 500 microL of 0.4 M sodium borate buffer, pH 8.3, containing 0.9 M NaC1 at 37 degrees C that reduced the subestimation error to less than or equal to 3% (rat serum) and less than or equal to 0.1% (human serum) and increased the ACE activity twofold to threefold. The Km and Vmax are reported for rat serum ACE (Hip-His-Leu) and dipeptidase (His-Leu) in borate buffer and phosphate buffer. Rat serum ACE hydrolysis of Hip-His-Leu measured by fluorometry correlated (r = 0.99, p less than 0.05) with the hydrolysis of angiotensin I measured by high-performance liquid chromatography. A direct method based on amino acid analysis is described for evaluating the dipeptidase error of complex mixtures such as tissue extracts and other physiological fluids. We have found that the assay can be used to measure ACE activity in 25 samples (in duplicate) in 2 hours with small intraassay (2.2%) and interassay (3.9%) coefficients of variation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984119 TI - Onset and dose relationships of ACTH effects on blood pressure in sheep. AB - The threshold and dose-response relationships for the blood pressure and metabolic effects of adrenocorticotropic hormone (corticotropin, ACTH) were examined in conscious sheep. Corticotropin was infused at five rates (0.5 micrograms/kg/day, n = 4; 1 micrograms/kg/day, n = 4;2 micrograms/kg/day, n = 6; 5 micrograms/kg/day, n = 5; and 10 micrograms/kg/day, n = 5) for 3 days, and the time of onset of the rise in blood pressure was assessed with a computer-based system. The effects of equimolar infusion of beta-endorphin and ACTH at 5 micrograms/kg/hour also were examined. Corticotropin infusion at 0.5 microgram/kg/day had no effect on mean arterial pressure. An ACTH infusion of 1.0 microgram/kg/day significantly increased mean arterial pressure (p less than 0.001), but the rise was less than that at the three higher doses, all of which produced similar effects. Changes in heart rate were significant at the 10 micrograms/kg/day level only (p less than 0.01). Initial urinary sodium retention was present at the three higher but not the two lower rates of infusion. Corticotropin infusion had no effect on urinary potassium excretion at any rate but produced hypokalemia at rates of 1.0 microgram/kg/day and above, which appeared to be dose related. Plasma sodium concentration was increased significantly only at the three higher rates (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984120 TI - Cyclic guanosine monophosphate mediates vascular relaxation induced by atrial natriuretic factor. AB - The biochemical mechanism of action of synthetic atrial natriuretic factor (atriopeptin II) was studied in vascular smooth muscle of the rabbit thoracic aorta. Atriopeptin II caused a time-dependent and concentration-dependent increase in tissue levels of cyclic guanosine monophosphate that corresponded in these same tissues with vascular relaxation. The elevation of arterial cyclic guanosine monophosphate levels preceded the onset of vascular relaxation. Atriopeptin II did not alter vascular levels of cyclic adenosine monophosphate. The presence of a functionally intact vascular endothelium was not necessary for atriopeptin II to elicit vascular relaxation. Atriopeptin II-induced vascular relaxation and elevation of cyclic guanosine monophosphate levels were inhibited by the guanylate cyclase inhibitor methylene blue. These data suggest cyclic guanosine monophosphate mediates vascular relaxation produced by atriopeptin II. PMID- 2984121 TI - Role of myeloperoxidase in respiratory burst of human polymorphonuclear leukocytes. Studies with myeloperoxidase-deficient subjects. AB - A comparative study of the respiratory burst [monitored as superoxide (O2-) production] of normal and myeloperoxidase (MPO) -deficient polymorphonuclear leukocytes (PMNs) was carried out on 11 MPO-deficient subjects that represent the largest sample of this kind ever studied. The rate of O2- production by isolated PMNs and whole blood from normal and MPO-deficient subjects was comparable during the initial 30-40 min of incubation with serum-treated zymosan (STZ). Afterwards, the amount of O2- produced became progressively higher in MPO-deficient cells at least until 120 min incubation with STZ. On the contrary the rate of O2- production by both cell types in response to 4-beta-phorbol-12-myristate-13 acetate (PMA) was the same. The PMNs of four MPO-deficient subjects were tested for their ingestion ability by counting the number of ingested particles on toluidine blue-stained sections of epoxy-embedded PMN suspensions. Both cell types ingested STZ particles at a comparable rate at early postphagocytic times, whereas on prolonged incubation MPO-deficient PMNs ingested more STZ particles than normal PMNs. These results suggest that the ingestion capacity of normal cells may undergo a more rapid deterioration than that of MPO-deficient cells during incubation with STZ. Evidence for a higher deterioration of normal PMNs with respect to MPO-deficient PMNs was obtained also from studies on the effect of storage on O2- generation. After standing at melting ice temperature for 3 h, normal PMNs produced less O2- than MPO-deficient PMNs in response to PMA, and the difference in O2- production by the two cell types in response to STZ was evident at earlier postphagocytic periods than with freshly isolated cells. Taken all together these results suggest that normal PMNs and MPO-deficient PMNs do not intrinsically differ in O2- generating potential and that the difference in the respiratory burst observed during phagocytosis may be accounted for by a more marked deterioration, in normal PMNs, of one or more functions related to the respiratory burst. PMID- 2984122 TI - Stimulus interactions in release of superoxide anion (O2-) from human neutrophils. Further evidence for multiple pathways of activation. AB - A wide variety of agents stimulate superoxide anion (O2-) release from human neutrophils. To determine whether the same or different cellular pathways are utilized, neutrophils were stimulated to release O2- with combinations of f-Met Leu-Phe (FMLP) (10(-7)M), C5a (25 nM), Con A (100 micrograms/ml), arachidonic acid (100 microM), and PMA (1 microgram/ml). These concentrations produced maximal O2- production when used alone. A synergistic response was observed when Con A was used in combination with FMLP or C5a. This response was twice the expected release in cytochalasin B-treated cells and three to five times the expected release in untreated cells. Additional studies showed that synergism was dependent upon the simultaneous presence of both agents. Additive O2- responses were observed when either FMLP, Con A, or C5a was tested in combination with arachidonic acid and when FMLP and C5a were tested together. When PMA was tested with C5a, FMLP, or Con A, a nonadditive O2- response resulted, whereas mixtures of PMA and arachidonic acid resulted in a less than additive response. These contrasting results using different soluble stimuli in combination suggest that multiple pathways exist for the stimulation of neutrophil O2- release, with some stimuli being totally independent of each other (possibly activating separate pools of oxidase), while other stimuli show cooperative effects on oxidase activation. PMID- 2984123 TI - Environmental silica in badger lungs: a possible association with susceptibility to Mycobacterium bovis infection. AB - Badger lungs contain dark granular foci (0.2 to 2.0 mm) comprising aggregates of enlarged macrophages containing birefringent crystalline particles. Particles were examined from the lungs of three badgers; many were silicates and a significant number were pure silica (SiO2). The particles and the accompanying pathology resembled mixed dust fibrosis and silicosis in humans, diseases associated with increased susceptibility to tuberculosis. PMID- 2984124 TI - Distinct bacteriocin groups correlate with different groups of Streptococcus mutans plasmids. AB - A correlation between the presence of 5.6-kilobase plasmids and bacteriocin activity was found in human-derived strains of Streptococcus mutans. Compared with bacteriocin activity of randomly selected clinical isolates of plasmid negative strains, bacteriocin activity of plasmid-positive strains significantly inhibited not only two laboratory strains of S. mutans, OMZ176 and AHT (P less than 0.0001 and P = 0.038, respectively), but most plasmid-negative clinical isolates as well (P = 0.0005). Comparisons of inhibition between pairs of plasmid positive strains revealed two groups, group I and group II, that produced distinct bacteriocins we designated as mutacin I and mutacin II, respectively. Within each group, a strain produced inhibitory activity against all the members of the other group but against no members of its own group. Plasmid DNA from plasmid-positive strains of each group was isolated and analyzed by restriction endonuclease digestion. Plasmids from the two groups that were apparently identical in size differed in digestion patterns for EcoRI, HaeIII, and TaqI, even though six TaqI fragments appeared to be common to all. Based on bacteriocin profiles and restriction enzyme digests, two distinct groups of plasmid-positive S. mutans strains emerged. Although the bacteriocin activity of plasmid-positive strain LM7 (serotype e) placed it in group I, clear differences in restriction digests distinguished it from the other plasmid-containing strains. PMID- 2984125 TI - Substitution reactions of 99mTcNCl-4--A route to a new class of 99mTc radiopharmaceuticals. AB - The preparation of sodium tetrachloronitridotechnetate Na[99mTcNCl4] is described. This complex may be used for the preparation by a substitution route of 99mTc-radiopharmaceuticals containing a Tc-nitrido(TcN) group. The labelling procedure was demonstrated by the preparation of [99mTcN]MDP, [99mTcN]DTPA and [99mTcN]cysteine. Biological distribution studies in mice of these complexes showed that the presence of the nitrido group produced a radiopharmaceutical with different biological behaviour to that produced when conventional reduction methods are used with the same ligand. HPLC studies confirmed that the use of the 99mTcNCl-4 substitution route produced radiopharmaceuticals of high radiochemical purity. PMID- 2984126 TI - Phosphatidylcholine synthesis in adult Dirofilaria immitis females. PMID- 2984127 TI - Radiation-induced decomposition of the purine bases within DNA and related model compounds. AB - This survey focuses on recent developments in the radiation chemistry of purine bases in nucleic acids and related model compounds. Both direct and indirect effects of ionizing radiation are investigated with special emphasis on the structural characterization of the final decomposition products of nucleic acid components. Available assays for monitoring radiation-induced base lesions are critically reviewed. PMID- 2984128 TI - Characterization of hydroxyl radical-induced damage after sparsely and densely ionizing irradiation. AB - The extent of hydroxyl radical mediated cell inactivation was measured for a variety of particle beams ranging from 8.5 Me V/u neon ions to 570 Me V/u argon ions. In general, the fraction of the total radiosensitivity caused by OH decreases from close to 60 per cent at low ionization density or low linear energy transfer (low LET) to close to 25 per cent at high LET for aerobically irradiated mammalian cells. The extent of OH induced cell lethality can be explained in terms of LET infinity only for low energy or low atomic number particles where fragmentations and complicated track structures do not contaminate the characteristic particle LET. For example, at a calculated LET infinity of 100 ke V/micron, the OH mediated fraction of the total radiation damage is about 25 per cent for low energy carbon but close to 40 per cent for high energy carbon ions. For low energy charged nuclei of approximately the same energy, as the 5.4-13.4 MeV/u He, Li, C and Ne ions in this report, there is a predictable diminution of the OH mediated effect with increasing LET infinity; however, the biological effect cannot be predicted accurately from calculated LET infinity values for high energy particle irradiation, nor indeed from a variety of low energy charged particles of quite different energies (incident velocities). This illustrates the unsuitability of using LET as a unifying parameter, except under specific circumstances. As more is learned about the energy deposition for energized charged particles in terms of track structure (core and penumbra), it may be possible to characterize the radiobiological data with a better physical parameter than LET infinity. PMID- 2984129 TI - WR-2721 inhibition of radiation-induced prostaglandin excretion in rats. AB - Pre-irradiation administration of the radioprotectant drug WR-2721 to rats resulted in a significant reduction in radiation-induced increases in excretion rates of prostaglandins (PGE and PGF2 alpha) and thromboxane (TxB2). In animals not irradiated. WR-2721 did not significantly alter these excretion rates. Dramatic reductions in the levels of urinary PGE and TxB2 were observed following exposure to 9.0 Gy of whole-body, unilateral gamma-radiation in WR-2721-treated animals, whereas changes in PGF2 alpha levels were less pronounced. Radiation induced diuresis was also significantly depressed in animals given WR-2721 before irradiation. Reduced prostaglandin excretion rates may reflect the general radioprotective capacity of the chemoprotector WR-2721 on the release of prostaglandins from radiation-damaged tissue. The decrease in diuresis may be related to the observed prostaglandin decreases. PMID- 2984130 TI - Reaction rate of OH radicals with phi X174 DNA: influence of salt and scavenger. AB - A derivation is given for the dependence of the rate constant of the reaction of OH radicals with a spherical macromolecule on the rate by which such radicals are scavenged by the medium. Experiments were carried out with oxygenated solutions of dilute single-stranded phi X174 DNA at 10(-4)M NaCl (large reaction radius of DNA) or at 10(-4)M NaCl + MgCl2 (small reaction radius) with t-butanol as a scavenger. The results of these experiments cannot be described by simple second order competition, but can be explained by the predicted dependence of the rate constant of the reaction OH + DNA on the concentration of t-butanol. Furthermore, the results show that only part of the reactions of OH radicals with phi X174 DNA leads to DNA inactivation, and that even at zero scavenger concentration OH radicals are scavenged by other molecules than DNA, presumably impurities remaining even after careful purification of the DNA. PMID- 2984131 TI - [Diagnosis of adenovirus infections]. PMID- 2984132 TI - [AIDS (acquired immunodeficiency syndrome)]. PMID- 2984133 TI - [Significance of genetic technics for the diagnosis and prevention of hepatitis B virus infections]. PMID- 2984134 TI - [Latent persistent infection caused by herpes simplex and varicella zoster virus]. PMID- 2984135 TI - Serial transplantation of a heterologous, mixed Mullerian tumor in the nude (athymic) mouse: growth characteristics and morphology. AB - In view of the poor prognosis for patients with mixed Mullerian tumors, transplantation of these tumors into nude mice offers an important model for the study of the physiologic processes occurring in this type of malignancy. We report the growth and serial transplantation of all elements of this tumor in the nude mouse. The lag period for establishment of the original tumor was 5 weeks. The tumor required 11 weeks to achieve a 2 X 2 X 1 cm volume at the initial pass, but on subsequent passes reached this volume in 6-12 weeks (mean = 8.3). Microscopic examination revealed that the epithelial, stromal and heterologous elements were serially transplanted. An osteoid matrix, confirmed by special staining for bone, was identifiable at both the light and electron microscopic levels. Throughout successive passages, the tumor retained the morphologic features of the original tumor. This tumor line is stable and reproducible, providing a readily available source of this ovarian carcinoma for clinical research studies. PMID- 2984136 TI - Rabbit corneal endothelial cell surface glycoproteins. AB - Sialic acid residues of plasma membrane glycoproteins of rabbit corneal endothelial cells were radiolabeled by oxidation with sodium periodate and reduction with sodium borotritide. Surface-labeled glycoproteins were resolved by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The major surface labeled glycoproteins were designated GP 1-8 in order of their increasing mobility on the gel (M.W. = 220K (GP-1), 200K (GP-2), 170K (GP-3), 135K (GP-4), 110K (GP-5), 95K (GP-6), 80K (GP-7), and 44K (GP-8). On the basis of the behavior of these glycoproteins on various carrier-bound lectins, preliminary information concerning their saccharide moieties was obtained. All 8 components bound to agarose-linked wheat germ agglutinin; GP 4-6 bound to concanavalin A and GP 6-7 bound to Ricinus communis agglutinin. No component bound to Bandeiraea simplicifolia I, Bandeiraea simplicifolia II, Ulex europeus or soybean agglutinin. These data suggest that in addition to the presence of sialic acid/N acetylglucosamine residues in all the eight glycoproteins, oligosaccharides with terminal beta-galactose residues occur in GP-6 and GP-7 while mannose (glucose) residues occur in GP 4-6. PMID- 2984137 TI - Studies of ocular murine cytomegalovirus infection. AB - The pathogenesis of ocular cytomegalovirus (CMV) infection in mice was studied in detail as a model of ocular involvement of human CMV infection. Smith strain of mouse CMV (MCMV) was inoculated into the anterior chamber of the eye and viral antigen was located by immunofluorescence. When salivary gland passaged MCMV (SG MCMV) was inoculated into the 12- to 18-day-old ICR/Sic mice, it elicited transient uveitis, retinitis, and scleritis during the early phase of infection followed by spread into the lacrimal glands, extraocular muscles and salivary glands. Mouse embryonic fibroblasts (MEF) passaged attenuated MCMV (CC-MCMV) caused mild uveitis with a short duration even inoculated into young mice, and the viral antigen was detected only in salivary glands thereafter. SG-MCMV titer in the eye taken from the young mice decreased from 10(4.5) TCID50/0.1 g at 2 days postinfection (PI) to 10(3.0) TCID50/0.1 g at 21 days PI, whereas in the salivary glands, it became detectable at 5 days PI with a titer of 10(5.0) TCID50/0.1 g, which increased up to 10(8.7) TCID50/0.1 g at 21 days PI. CC-MCMV was detectable only in the young mice salivary glands ranging from 10(4.7) TCID50/0.1 g at 9 days PI to 10(7.3) TCID50/0.1 g at 21 days PI. When the lens capsule had been damaged during the inoculation procedure, the mice developed cataracts with evidence of viral growth in the lens. When SG-MCMV was inoculated onto the cornea, viral antigen was not detected during the period of the present experiment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984138 TI - Suppression of secondary herpes simplex uveitis by cyclosporine. AB - The authors studied the effect of an immunosuppressive agent, cyclosporine (CyA), on experimental secondary herpes simplex (HS) uveitis. Secondary HS uveitis was induced in a rabbit eye that had recovered from primary HS uveitis by challenging it with an intravitreal injection of herpes simplex virus (HSV) antigen. Daily intramuscular injections of CyA (25 mg/kg body weight) for 7 days prior to the intravitreal challenge with HSV antigen significantly suppressed the induction of secondary HS uveitis, but daily injections of CyA after the challenge with HSV antigen was ineffective. Intravitreal injections of CyA (5 mg) 7 days and 3 days prior to the HSV challenge were less effective, but the combined treatment with seven daily intramuscular CyA and two intravitreal CyA injections prior to the HSV challenge was most effective in the prevention of the uveitis. The daily intramuscular treatment with CyA resulted in a marked reduction of cell-mediated immunity while leaving the level of circulating HSV specific antibody high. No reactivation of latent HSV was detected in trigeminal and superior cervical ganglia of CyA-treated rabbits. PMID- 2984139 TI - Latent and active plasminogen activator in corneal ulceration. AB - Previous studies of alkali burns have provided evidence for an important role of the plasminogen activator (PA)/plasmin system in corneal ulceration. Current studies have utilized a sensitive, plasminogen-dependent fluorescent assay to demonstrate that PA is present mostly in a latent (trypsin- or plasmin activatable) form (proactivator) in cultures of rabbit corneal epithelial cells or normal corneas. Cultures of ulcerating corneas demonstrate only active PA early in organ culture, whereas, latent PA levels increase later in culture. Thus, ulceration is correlated with the apparent conversion of latent to active PA. Moreover, profiles of proactivator and latent collagenase and of active PA and active collagenase in vitro, respectively, are similar, suggesting that activator and collagenase are under coordinate control. Cultures of normal epithelial cells and nonulcerating corneas contain PA molecular weight species of 72,000 and 46,000 MW, and ulcer corneas, species of 72,000, 46,000, and 35,000 MW. Double-diffusion analysis indicates that rabbit epithelial cells, fibroblasts, and ulcer corneas produce urokinase (UK)-like PA; and human cornea extracts and tears also contain PA immunoreactive with anti-UK antibodies. The existence of PA in a latent form identifies another level of regulation in the cascades that lead to stromal ulceration. PMID- 2984140 TI - Spontaneous ocular shedding of HSV-1 in latently infected rabbits. AB - The unscarified corneas of rabbits were inoculated with 50 microliter of 2-4 X 10(6) PFU/ml of herpes simplex virus, type 1 (HSV-1), McKrae strain in 10 separate experiments over a 12-month period. Sixty of 104 (57.7%) rabbits survived to postinoculation (PI) day 20. These sixty rabbits were swabbed with dacron-tipped swabs for twenty consecutive days (PI days 20-39). The tear film collected on the swabs was immediately placed in tissue culture tubes with confluent primary rabbit kidney (RK) cell monolayers. The RK monolayers were monitored for cytopathic effects indicative of HSV-1. Fifty-eight of the sixty rabbits (96.7%) inoculated had at least one positive episode. Ninety-three of the 120 eyes (77.5%) of the latently infected rabbits had at least one positive episode. Virus was detected in 72 of the 93 positive eyes (77.4%) between PI days 20 and 29 and in 21 of the 93 positive eyes (22.5%) between PI days 31-39. A total of 2400 swabs were taken and 324 were positive (13.5%). All of the 58 positive rabbits were used later for ocular induction of HSV-1 and all 116 eyes of the latently infected rabbits shed virus for at least four consecutive days during induction. PMID- 2984141 TI - CT evaluation of hepatic and splenic trauma with EOE-13. An experimental study in monkeys. AB - EOE-13 is a liver-spleen specific CT contrast agent currently undergoing clinical trials. It is an aqueous emulsion of iodinated fatty acids. We studied eight rhesus monkeys with surgically created blunt trauma to the liver and spleen. The animals were examined using CT scans without contrast material and CT scans with EOE-13. Scans were performed at times between 3 hours and 24 hours after trauma. The presence of parenchymal injury was shown better EOE-13 (P less than 0.01). The use of EOE-13 permitted earlier diagnosis in both the liver and spleen. This contrast agent has potential for the evaluation of trauma in clinical practice, and may be particularly useful in children. PMID- 2984142 TI - Comparison of four methods for typing low-passage herpes simplex virus isolates. AB - Clinical isolates of herpes simplex virus (HSV) were identified as HSV type 1 or type 2 by sensitivity to (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), by differential replication in chick embryo cells versus guinea pig embryo cells, by restriction endonuclease analysis, and by a direct fluorescent antibody technique using monoclonal antibodies. More than 550 isolates were typed by two or three of the systems with complete agreements as to virus type between systems for each isolate. In appropriately equipped laboratories, any of the above typing systems can be used with complete confidence. However, the BVDU sensitivity assay, particularly when used in a continuous cell line as described, can be economically utilized in any virology laboratory. PMID- 2984143 TI - Oncogene expression in isogenic, EBV-positive and -negative Burkitt lymphoma cell lines. AB - To elucidate the mode of action of Epstein-Barr virus (EBV), we have examined the possibility that EBV infection may lead to a change in oncogene expression. Levels of oncogene-related RNA were compared in the Burkitt lymphoma(BL)-derived, EBV-negative cell line BJAB and in two isogenic, EBV-converted sublines of BJAB whose growth characteristics differ from the EBV-negative parent line. The oncogenes c-myc, Ha-ras, and Blym were each found to be expressed at equally high levels in the three cell lines. This suggests that the mechanism of action of EBV in altering the growth properties of lymphoid cells is not dependent on the induction of cellular oncogenes. In addition, since the 8,14 chromosomal translocation has been related to increased c-myc expression in BL cells, a karyotype analysis was done to confirm the initial report of a normal karyotype in BJAB cells. Chromosomes 8 and 14 were seen to be normal. Thus, BJAB cells are a BL cell line in which the elevated expression of c-myc is independent not only of EBV but also of translocations involving chromosome 8. PMID- 2984144 TI - Morphology and morphogenesis of viruses of hemorrhagic fever with renal syndrome (HFRS). I. Some peculiar aspects of the morphogenesis of various strains of HFRS virus. AB - The morphology and morphogenesis of viruses of hemorrhagic fever with renal syndrome (HFRS) were studied by thin-section electron microscopy. Ten strains of HFRS virus isolated in China and one strain isolated in Korea were compared. The virions varied in size as well as in the shape and character of their inner components. Both intracellular and extracellular ribosome-like, electron-dense, virus-associated granules were seen. A viral antigen layer was often visualized on the surface of HFRS virus-infected cells, as confirmed by immune electron microscopy. Aberrant tailed particles and elongated rod-like particles were seen infrequently. The morphogenesis of HFRS viruses appears to be more complex than that of other members of the family Bunyaviridae. The prototype Hantaan virus shares a common morphology and morphogenesis with the other HFRS viruses studied. PMID- 2984145 TI - Cholecalciferol metabolites in polycythemic newborns. AB - Metabolites of cholecalciferol were measured in the sera of healthy and polycythemic newborns at 15.5 and 10.5 h of age (median), respectively. The serum concentrations of 1,25-dihydroxyvitamin D and 24,25-dihydroxyvitamin D were lower in the polycythemic group (P less than 0.003 and P less than 0.001, respectively). As the kidney is the main site of conversion of 25-hydroxyvitamin D to its dihydroxylated metabolites, it is possible that hyperviscosity and subsequent decrease in renal blood flow interfere with the efficiency of the process. PMID- 2984147 TI - The radiological impact of mining in a Th-rich Norwegian area. AB - The radiological impact of former mining activities in the Fen area in southern Norway has been assessed. The area is known to have enhanced concentrations of Th. A recent epidemiological study suggested a significant increment of lung cancer among the former miners, and the doses to the miners have been assessed. The main contributor to the effective dose equivalent in one of the mines was inhaled 220Rn progeny. Water in drilled wells and in nearby lakes has been analysed for 220Rn and Ra. Uranium and Th analyses were performed on mine and lake water. The mining activity does not seem to have contaminated drinking water significantly. The tailings from Nb production has enriched Ra and Th concentrations. The tailings and the possible use of waste rock from the mining are probably the most important environmental results of the mining activities. PMID- 2984146 TI - Theoretical models for determining 222Rn and 220Rn progeny levels in Canadian underground U mines--a comparison with experimental data. AB - Use has been made of several theoretical models to predict radiation levels in underground U mines. The models used are the Evans model, the Thomas-Epps mine model, the isolated mine model, the mine tunnel with no air flow and the mine tunnel with air flow (Beckman and Holub). Calculations based on the above models have been extended to include 220Rn gas and its progeny, a common occurrence in some Canadian U mines. Theoretical predictions include 222Rn and 220Rn progenies working levels and concentrations, as well as some ratios of great practical interest. The main differences between the models are pointed out and comparison is made with experimental data gathered during the last 3 yr in several Canadian underground U mines. In general terms, the mine models reported in the literature and presented here are neither satisfactory nor clearly distinguishable for practical application on the basis of the experimental data so far collected. The reason for such lack of agreement lies mainly in the unrealistic assumptions on which the models are based. Compounded is the gross oversimplification of quite complex dynamic situations encountered in actual practice. Deficiencies inherent to the models are noted and suggestions to improve the applicability of mine models to practical situations are indicated. PMID- 2984148 TI - Scattered radiation doses to some critical organs during pediatric radiotherapy. AB - The levels of scattered radiation doses imparted to the eyes, thyroid and gonads of pediatric patients treated with orthovoltage radiation (300 kVp, 2.0 mmCu HVL) and with a 4-MV linear accelerator, were determined by making thermoluminescent dosimeter (TLD) measurements in three paraffin phantoms of different sizes. These phantoms were made from molds of mannequins used for store display, of approximate heights 30", 40" and 50", representing children of ages 1-2, 4-5 and 8-10 yr, respectively. The sites chosen for irradiation were (1) the whole brain, (2) the chest, (3) the kidney bed, (4) the whole abdomen and (5) the spinal column. These sites are normally treated in such pediatric malignancies as medulloblastoma, neuroblastoma and Wilms' tumor. Some of the doses measured are less than 10 rad for an entire treatment regimen, and would therefore be categorized as low-level doses. Where radiation was the only mode of treatment for long-term survivors of such malignancies, especially those treated 20-30 yr ago with orthovoltage radiation, useful data may be extracted for contributing to our knowledge about the long-term effects of low levels of radiation. PMID- 2984149 TI - Liver carcinoma in an athlete taking anabolic steroids. PMID- 2984150 TI - Defeminizing ovarian tumor: report of a case. PMID- 2984151 TI - Fast neutrons and misonidazole for malignant astrocytomas. AB - Twenty-five patients with biopsy proven malignant supratentorial astrocytomas were entered into a Phase I/II study of misonidazole combined with neutron radiation at Fermilab Neutron Therapy Facility (NTF) between August 1979 and April 1981. The main objectives were to determine tissue tolerance in terms of acute and late effects, and to estimate tumor clearance and survival rates. The total dose was 18.0 Gy given in weekly fractions of 3.0 Gy over 39 days. Four hours before each irradiation, 2.5 gm/m2 misonidazole was administered orally. Patients' ages ranged from 28-69 years. Karnofsky status for most patients was 80 or 90; the lowest grade was 60. The majority of patients had glioblastoma multiforms. Most were already on steroids prior to initiation of therapy. The median survival for the whole group was 12.0 months; 25% were alive at 18 months with some neurological compromise. The median survival remained unchanged for subgroups of patients with ages between 40-60 years and with Karnofsky performance status above 80. Among the 19 patients with glioblastoma multiforme, the median survival was 10 months. Acute toxicity was within tolerable limits. Details of toxicity and tissue analysis from post mortems and second craniotomy samples are presented. PMID- 2984152 TI - Radiation therapy for pancreatic cancer: eleven year experience at the JCRT. AB - Radiation therapy (XRT) for 41 patients with unresectable pancreatic cancer resulted in a median survival of 7.0 months. There was no difference in median survival for patients receiving external beam alone (3500 to 5600 cGy) (n = 28), intraoperative (IORT) boost plus external beam (5040 to 6750 cGy) (n = 9), or a gold-198 implant +/- external beam radiation (n = 4). A pilot study using orthovoltage IORT boost indicates no acute toxicity with doses of 1250 to 1750 cGy. Serious late damage has not been observed in any patients followed to 2 years. Local recurrence in patients treated post-operatively after "radical" surgery occurred in one of 10 (10%). This adjuvant treatment is safe and appears to improve local control rates compared to historical data, but survival is still poor. The median survival for the post-operative group is 10 months; three patients are alive without disease 8 months to 8.3 years after treatment. PMID- 2984153 TI - Production and metabolism of lignans by the human faecal flora. AB - Lignans have, until recently, been found only in plants. Enterolactone and enterodiol are the major lignans present in the urine of humans and have a potential physiological protective role against cancer. It has been shown that these compounds can be formed in vitro by human faecal flora and that enterodiol is oxidized to enterolactone by bacteria that are present in stools at a concentration of up to 10(3)/g. It was also possible to produce both of these lignans in vitro from linseeds and from secoisolariciresinol, a precursor present in linseed, by bacteria present in stools, at a concentration of between 10(3) and 10(4)/g. Enterolactone was produced from matairesinol, a more abundant plant lignan than secoisolariciresinol, after incubation with a mixed faecal flora under both aerobic and anaerobic conditions. In each case conversion was dependent on the presence of viable bacteria. These findings indicate that a number of different pathways operate to produce enterolactone and enterodiol depending on the ingested dietary precursor. PMID- 2984154 TI - The scope and limitations of four methods for the rapid identification of Enterobacteriaceae in foods. AB - Four methods for the identification of Enterobacteriaceae in cooked frozen meat have been appraised. Although these methods could be modified to improve efficiency they could be clearly classified into two categories: (i) those that allow identification to genus level and which often cannot discriminate between two genera without the help of additional reactions; (ii) those that allow identification to species level and which only occasionally require additional reactions to differentiate one species from another. A different evaluation criterion was followed for each of these categories. In the first, as methods lead to final identification by means of data furnished by the manufacturer or via a number code which does not indicate probability, reactions were compared with those obtained by classical methods. In the second group, every number code admits three possibilities and indicates a probable biotype for each, thus reducing the risk of faulty interpretation on the part of the operator while also compensating for deficiencies of certain reactions. Evaluation was therefore based on the percentage of correctly identified cases per species. Efficiencies of the main biochemical reactions are also discussed. PMID- 2984155 TI - A new beta-lactamase plasmid in Neisseria gonorrhoeae. PMID- 2984156 TI - The use of bacteria producing the aminoglycoside inactivating enzyme ANT-(2") in an in-vitro model. AB - A series of transconjugants of Escherichia coli W3110 containing different plasmids coding for the aminoglycoside inactivating enzyme ANT-(2") were tested in an in-vitro model simulating the concentration time curves of aminoglycosides after intramuscular administration of 80 or 120 mg. There was good correlation of the MIC, which ranged from 2 to 32 mg/l, with the killing ability of the drugs. A second dose had no or only little effect on ANT-(2") producing strains. All strains, although of 'intermediate' sensitivity were found to be resistant in the in-vitro model. Only netilmicin could eliminate some of the strains but its activity on ANT-(2")-positive strains was not as good as on ANT-(2")-negative ones. PMID- 2984157 TI - Effect of cutaneous denervation of face and trunk on thermoregulatory responses to cold in rats. AB - The effect of eliminating afferent input from cutaneous thermoreceptors of the face and trunk on the ability of rats to regulate body temperature in cool environments was studied. Thermoregulatory ability in a cool environment was assessed first in a 25 degrees C environment and then during slow (20 min) and rapid (5 min) reductions of ambient temperature (Ta) to 15 degrees C by monitoring rate of heat production, rectal temperature, and skin temperature on the back, ear, and tail. These measurements were made in four rats while they were intact and during the 2 wk after cutaneous denervation. Rats were found to regulate body temperature well even after the cutaneous nerves of the trunk and face were sectioned. In eight rats the metabolic curve was determined before and 7-10 days after cutaneous denervation. Although the minimal resting metabolic rates did not differ in the two conditions, the lower critical temperature was significantly elevated from 26.8 to 28.9 degrees C and the rate of rise in metabolic rate per degrees celsius decrease in Ta was also significantly higher after cutaneous denervation. It appears that the higher rate of heat production is in compensation for an increase in the rate of heat loss in denervated rats. PMID- 2984158 TI - Chronic exercise and left ventricular structure and function in healthy human subjects. AB - Twelve healthy well-trained participants in a supervised exercise program (mean age, 41.3 yr) were compared with 12 sedentary control subjects (mean age, 38.9 yr) with physical characteristics similar to the exercised group (EG) before training. Resting echocardiograms revealed significantly lower heart rates (HR) in the EG compared with control group (CG) but no evidence for cardiac structural differences between groups. Radionuclide angiograms performed at rest and during two levels of supine cycling (HR targets: 120 and 140 beats X min-1) resulted in increases in background-corrected end-diastolic counts [EDC(bc)] and confirmed use of the Frank-Starling mechanism in the majority of subjects. Mean values (+/- SD) for ejection fraction (EF) and normalized peak systolic ejection rate (PSER) (P greater than 0.05 between groups) were the following. (Formula: see text) The results suggested that fitness training does not induce significant cardiac enlargement as apparent from measurements at rest or important changes in contractile state during exercise. Increases in exercise stroke volume with such training may be the result of an increased end-diastolic volume. PMID- 2984159 TI - On the relation between expiratory duration and subsequent inspiratory duration. AB - To characterize respiratory interphase relationships in dogs, inspiratory duration (TI) or expiratory duration (TE) was systematically altered by electrical activation of vagal afferents, and the effect on subsequent TE or TI values was measured from the phrenic discharge. A linear TI-subsequent TE relationship was found. Following a vagally mediated prolongation of TE, 1) TI was prolonged, and the TE-subsequent TI relationship was curvilinear, 2) the threshold for inspiratory termination by phasic vagal inputs was increased, 3) the amplitude of the time course of the phrenic discharge was reduced so that the peak discharge reached the same level at inspiratory termination independent of TI, 4) the effect on TI prolongation persisted for several breaths whereas the effect was minimal on subsequent TE values, and 5) for tonic inputs the direct shortening of TI was nearly offset by the indirect lengthening of TI. These studies suggest the existence of slow central mechanisms that provide adaptation to elevated levels of vagal input in the control of TI. These mechanisms may also be responsible for the interphase timing relationships. PMID- 2984160 TI - Epinephrine is unessential for stimulation of liver glycogenolysis during exercise. AB - To determine the role of adrenal medullary hormones in controlling the rate of liver glycogenolysis during exercise, adrenodemedullated (ADM) and sham-operated (SO) rats were run on a rodent treadmill at 21 m/min up a 15% grade for 0, 30, or 60 min. Rats were anesthetized by intravenous injection of pentobarbital sodium, and liver, muscle, and blood were collected and frozen. Liver glycogen decreased at similar rates in ADM and SO rats. Hepatic adenosine 3',5'-cyclic monophosphate (cAMP), plasma glucagon, and plasma free fatty acids increased to the same extent in both ADM and SO rats. The adrenodemedullation caused a reduction in glycogenolysis in the fast-twitch white region of the quadriceps, soleus, and lateral gastrocnemius during exercise. The normal exercise-induced increase in blood glucose and lactate and the decline in plasma insulin were not observed in the demedullated rats. During submaximal exercise the principal targets for epinephrine released from the adrenal medulla appear to be pancreatic beta-cells and skeletal muscle and not the liver. PMID- 2984162 TI - Effect of hypoxemia on the renin-angiotensin-aldosterone system in humans. AB - Hypoxemia was induced in five subjects older than 40 (group 1) and five younger than 35 yr (group 2) on normal and low-salt diets by having the subjects breathe hypoxic gas. The fractional inspired O2 of the hypoxic gas was regulated so that group 1 hemoglobin saturations fell to 90% for 1 h. Group 2 subjects had desaturation to 90% for 1 h followed by desaturation to 80% for a 2nd h. Plasma renin activity (PRA), angiotensin-converting enzyme activity (ACE), and plasma cortisol levels did not change during hypoxemia. Plasma aldosterone levels fell in both groups during the 1st h of hypoxemia. Decreases were greatest during salt restriction and were significant (P less than 0.01) for the combined groups. Plasma aldosterone levels plateaued during the 2nd h of more severe hypoxemia in group 2. Hepatic blood flow, measured by indocyanine green clearance, and the adrenal response to exogenous adrenocorticotropic hormone, measured by changes in plasma cortisol and aldosterone, were not changed by hypoxemia in group 2 subjects. These results indicate that plasma aldosterone falls during hypoxemia despite unchanged PRA, ACE, hepatic blood flow, and adrenal function. PMID- 2984161 TI - Carbon monoxide-cytochrome interactions in the brain of the fluorocarbon-perfused rat. AB - Reflectance spectrophotometry through the skull was used to investigate carbon monoxide (CO) binding by tissue hemoproteins in the brains of barbiturate anesthetized Sprague-Dawley rats. After splenectomy and extensive perfluorotributylamine exchange transfusion, steady-state spectral scans were obtained in Soret and visible wave-length regions during O2 ventilation, during subsequent exposure to O2-enriched gases containing 1, 3, or 5% CO, and finally after N2 anoxia. These CO exposures were well-tolerated and electroencephalograph (EEG) activity continued to be present. Initial difference spectra were influenced by CO binding to residual hemoglobin, but spectral evidence of CO mediated b-type cytochrome reduction was obtained in the visible region as CO concentration was increased to 3 or 5%. This was associated with Soret spectra compatible with formation of the reduced cytochrome a3-CO complex. Reduction of cytochrome a at 605 nm and cytochrome c + c1 at 550 nm was absent. These findings may indicate respiratory chain branching through b cytochromes, either to a separate a3-like oxidase independent of the classical cytochrome aa3 or to an unidentified alternative CO-sensitive oxidase. PMID- 2984163 TI - Model simulation of single-breath washout of insoluble gases from dog lungs. AB - Intrapulmonary gas mixing by convection and diffusion is analyzed on the basis of various analog lung models incorporating singly or in combination series inhomogeneity, parallel inhomogeneity, intercompartmental diffusion, sequential inspiration, and sequential expiration. The slope of the alveolar plateau of insoluble gases is used as an indicator for incomplete gas mixing. By use of the models, we have attempted to simulate experimental results obtained in artificially ventilated dog lungs with simultaneous single-breath washout of He and SF6 (Meyer et al., J. Appl. Physiol. 55: 1795-1802, 1983). Sufficient agreement with experimental data is obtained only when diffusion between serial parallel compartments and sequential expiration are incorporated, the first being mainly responsible for separation of He and SF6, the latter for the slope of the alveolar plateau of these gases. A combined operational model comprising two parallel compartments connected to a mixing compartment, with both convective and diffusive gas transport between the compartments, could account for most of the experimental observations. PMID- 2984164 TI - Cardiorespiratory reflexes from muscles during dynamic and static exercise in the dog. AB - Cardiorespiratory reflex responses during the initial phase of dynamic and static contraction of hindlimb muscles were studied in anesthetized dogs. Muscle contractions were elicited by stimulating the femoral and gastrocnemius nerves at 3 and 100 Hz with the intensity of 2.0-2.5 times the motor threshold for a 20-s period. Rhythmic contractions caused a decrease in arterial pressure (Pa) and heart rate (HR) and increased pulmonary ventilation (VE) by increasing frequency (f) without significantly changing VT. Tetanic contractions provoked an increase in Pa and HR and a hyperpnea resulting from a rise in both f and VT. Similar responses were also obtained in anesthetized dogs with carotid sinuses denervated and cervical vagi cut. The abrupt increase in VE at the start of both types of exercise was not associated with immediate significant decreases in end-tidal CO2 values. These two patterns of cardiocirculatory and respiratory responses were closely similar to those reported in anesthetized rabbits in previous studies. Both patterns of responses were reflexes initiated by activation of muscle receptors verified by interrupting the afferents from the contracting muscles. It is concluded that, during dynamic and static work, two distinct muscular reflex mechanisms might exert their drives, related to the muscular metabolic rate, on the circulatory and respiratory function. PMID- 2984165 TI - Modulation of cholinergic neurotransmission in airways by enkephalin. AB - We compared the effects of methionine enkephalin and leucine enkephalin on contractions of isolated canine tracheal smooth muscle strips induced by field electrical stimulation (ES) and exogenous acetylcholine (approximately 10(-5) M). Methionine and leucine enkephalin (10(-8) to 10(-5) M), when added at the peak of airway contractions induced by ES at 1 Hz, depressed the contractions in a concentration-dependent manner by a maximum of 95 and 99%, respectively. Acetylcholine-induced contractions of similar magnitude were depressed only 4% by methionine enkephalin and 12% by leucine enkephalin. Frequency-response curves (0.5-20 Hz) were also obtained before and after incubation of tracheal strips with 10(-5) M methionine and leucine enkephalin. Enkephalin depressed contractions induced by stimulation at 0.5 and 1 Hz by an average of 98 and 95%, respectively. The inhibitory effect of enkephalin progressively decreased at successively higher stimulus frequencies until at 20 Hz there was no significant difference between airway contractions obtained in the presence and absence of enkephalin. Naloxone (3 X 10(-5) M) antagonized the inhibitory effects of both enkephalins. We conclude that methionine and leucine enkephalins inhibit the release of acetylcholine from the postganglionic parasympathetic neurons that innervate airway smooth muscle. PMID- 2984166 TI - Inspiratory rhythm in airway smooth muscle tone. AB - In anesthetized paralyzed open-chested cats ventilated with low tidal volumes at high frequency, we recorded phrenic nerve activity, transpulmonary pressure (TPP), and either the tension in an upper tracheal segment or the impulse activity in a pulmonary branch of the vagus nerve. The TPP and upper tracheal segment tension fluctuated with respiration, with peak pressure and tension paralleling phrenic nerve activity. Increased end-tidal CO2 or stimulation of the carotid chemoreceptors with sodium cyanide increased both TPP and tracheal segment tension during the increased activity of the phrenic nerve. Lowering end tidal CO2 or hyperinflating the lungs to achieve neural apnea (lack of phrenic activity) caused a decrease in TPP and tracheal segment tension and abolished the inspiratory fluctuations. During neural apnea produced by lowering end-tidal CO2, lung inflation caused no further decrease in tracheal segment tension and TPP. Likewise, stimulation of the cervical sympathetics, which caused a reduction in TPP and tracheal segment tension during normal breathing, caused no further reduction in these parameters when the stimulation occurred during neural apnea. During neural apnea the tracheal segment tension and TPP were the same as those following the transection of the vagi or the administration of atropine (0.5 mg/kg). Numerous fibers in the pulmonary branch of the vagus nerve fired in synchrony with the phrenic nerve. Only these fibers had activity which paralleled changes in TPP and tracheal tension. We propose that the major excitatory input to airway smooth muscle arises from cholinergic nerves that fire during inspiration, which have preganglionic cell bodies in the ventral respiratory group in the region of the nucleus ambiguus and are driven by the same pattern generators that drive the phrenic and inspiratory intercostal motoneurons. PMID- 2984167 TI - Increasing gracilis muscle interstitial potassium concentrations stimulate group III and IV afferents. AB - Static muscular contraction reflexly increases arterial blood pressure and heart rate. One possible mechanism evoking this reflex is that potassium accumulates in the interstitial space of a working muscle to stimulate group III and IV afferents whose activation in turn evokes a pressor response. The responses of group III and IV muscle afferents to increases in interstitial potassium concentrations within the range evoked by static contraction are unknown. Thus we injected potassium chloride into the gracilis artery of anesthetized dogs while we measured both gracilis muscle interstitial potassium concentrations with potassium-selective electrodes and the impulse activity of afferents in the gracilis nerve. We found that increasing interstitial potassium concentrations to levels similar to those seen during static contraction stimulated 14 of 16 group III and 29 of 31 group IV afferents. The responses of the afferents to potassium were concentration dependent. The typical response to potassium consisted of a burst of impulses, an effect that returned to control firing rates within 26 s, even though interstitial potassium concentrations remained elevated for several minutes. Although our results suggest that potassium may play a role in initiating the reflex cardiovascular responses to static muscular contraction, the accumulation of this ion does not appear to be solely responsible for maintaining the pressor response for the duration of the contraction. PMID- 2984169 TI - Diffusion limitation in normal humans during exercise at sea level and simulated altitude. AB - The relative roles of ventilation-perfusion (VA/Q) inequality, alveolar-capillary diffusion resistance, postpulmonary shunt, and gas phase diffusion limitation in determining arterial PO2 (PaO2) were assessed in nine normal unacclimatized men at rest and during bicycle exercise at sea level and three simulated altitudes (5,000, 10,000, and 15,000 ft; barometric pressures = 632, 523, and 429 Torr). We measured mixed expired and arterial inert and respiratory gases, minute ventilation, and cardiac output. Using the multiple inert gas elimination technique, PaO2 and the arterial O2 concentration expected from VA/Q inequality alone were compared with actual values, lower measured PaO2 indicating alveolar capillary diffusion disequilibrium for O2. At sea level, alveolar-arterial PO2 differences were approximately 10 Torr at rest, increasing to approximately 20 Torr at a metabolic consumption of O2 (VO2) of 3 l/min. There was no evidence for diffusion disequilibrium, similar results being obtained at 5,000 ft. At 10 and 15,000 ft, resting alveolar-arterial PO2 difference was less than at sea level with no diffusion disequilibrium. During exercise, alveolar-arterial PO2 difference increased considerably more than expected from VA/Q mismatch alone. For example, at VO2 of 2.5 l/min at 10,000 ft, total alveolar-arterial PO2 difference was 30 Torr and that due to VA/Q mismatch alone was 15 Torr. At 15,000 ft and VO2 of 1.5 l/min, these values were 25 and 10 Torr, respectively. Expected and actual PaO2 agreed during 100% O2 breathing at 15,000 ft, excluding postpulmonary shunt as a cause of the larger alveolar-arterial O2 difference than accountable by inert gas exchange.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984168 TI - Ventilation-perfusion inequality in normal humans during exercise at sea level and simulated altitude. AB - To investigate the effects of both exercise and acute exposure to high altitude on ventilation-perfusion (VA/Q) relationships in the lungs, nine young men were studied at rest and at up to three different levels of exercise on a bicycle ergometer. Altitude was simulated in a hypobaric chamber with measurements made at sea level (mean barometric pressure = 755 Torr) and at simulated altitudes of 5,000 (632 Torr), 10,000 (523 Torr), and 15,000 ft (429 Torr). VA/Q distributions were estimated using the multiple inert gas elimination technique. Dispersion of the distributions of blood flow and ventilation were evaluated by both loge standard deviations (derived from the VA/Q 50-compartment lung model) and three new indices of dispersion that are derived directly from inert gas data. Both methods indicated a broadening of the distributions of blood flow and ventilation with increasing exercise at sea level, but the trend was of borderline statistical significance. There was no change in the resting distributions with altitude. However, with exercise at high altitude (10,000 and 15,000 ft) there was a significant increase in dispersion of blood flow (P less than 0.05) which implies an increase in intraregional inhomogeneity that more than counteracts the more uniform topographical distribution that occurs. Since breathing 100% O2 at 15,000 ft abolished the increased dispersion, the greater VA/Q mismatching seen during exercise at altitude may be related to pulmonary hypertension. PMID- 2984170 TI - Efficacy of high-frequency ventilation in presence of extensive ventilation perfusion mismatch. AB - Ten anesthetized normal dogs were each given two methacholine inhalational challenges to produce large amounts of low ventilation-perfusion (VA/Q) regions but little shunt. After one challenge, high-frequency ventilation (HFV) was applied, whereas after the other conventional mechanical ventilation (MV) was used, the order being randomized. Levels of both ventilatory modes were selected prior to challenge so as to result in similar and normal mean airway pressures and arterial PCO2 levels during control conditions. Gas exchange was assessed by both respiratory and multiple inert-gas transfer. Comparing the effect of HFV and MV, no statistically significant differences were found for lung resistance, pulmonary hemodynamic indices, arterial and mixed venous PO2, expired-arterial PO2 differences, or inert-gas data expressed as retention-excretion differences. The only variables that were different were mean airway pressure (2 cm higher during HFV, P less than 0.04) and arterial PCO2 (10 Torr higher during HFV, P less than 0.002). These results suggest that in this canine model of lung disease characterized by large amounts of low VA/Q regions, HFV is no more effective in delivering fresh gas to such regions than is MV. PMID- 2984171 TI - Cold sensitivity induced by overproduction of UmuDC in Escherichia coli. AB - The UmuD and UmuC proteins of Escherichia coli are essential for mutagenesis by UV and most chemicals. Their mode of action is presently unknown. Strains which lack the LexA repressor [lexA(Def)] and contain a pBR322-derived plasmid carrying the umuDC operon overexpress UmuD and UmuC and become cold sensitive (growth at 42 degrees C but not at 30 degrees C). Deletion mapping showed that the umuDC locus on the plasmid is responsible for conferring cold sensitivity. The conditional lethality appeared due to a rapid and reversible inhibition of DNA synthesis at the nonpermissive temperature. Cold sensitivity was enhanced by the increase of NaCl in the medium to 1% and eliminated by 4% ethanol in the medium. Cold sensitivity was partially suppressed by the lon-100 mutation and completely suppressed by the htpR165 mutation. PMID- 2984172 TI - Growth of Acinetobacter sp. strain HO1-N on n-hexadecanol: physiological and ultrastructural characteristics. AB - The growth of Acinetobacter sp. strain HO1-N on hexadecanol results in the formation of intracytoplasmic membranes and intracellular rectangular inclusions containing one of the end products of hexadecanol metabolism, hexadecyl palmitate. The intracellular inclusions were purified and characterized as "wax ester inclusions" consisting of 85.6% hexadecyl palmitate, 4.8% hexadecanol, and 9.6% phospholipid, with a phospholipid-to-protein ratio of 0.42 mumol of lipid phosphate per mg of inclusion protein. The cellular lipids consisted of 69.8% hexadecyl palmitate, 22.8% phospholipid, 1.9% triglyceride, 4.7% mono- and diglyceride, 0.1% free fatty acid, and 0.8% hexadecanol, as compared with 98% hexadecyl palmitate and 1.9% triglyceride, which comprised the extracellular lipids. Cell-associated hexadecanol represented 0.05% of the exogenously supplied hexadecanol, with hexadecyl palmitate accounting for 14.7% of the total cellular dry weight. Acinetobacter sp. strain HO1-N possesses a mechanism for the intracellular packaging of hexadecyl palmitate in wax ester inclusions, which differ in structure and chemical composition from "hydrocarbon inclusions" isolated from hexadecane-grown cells. PMID- 2984173 TI - Evidence that the outer membrane protein gene nmpC of Escherichia coli K-12 lies within the defective qsr' prophage. AB - Recombinants between phage lambda and the defective qsr' prophage of Escherichia coli K-12 were made in an nmpC (p+) mutant strain and in the nmpC+ parent. The outer membrane of strains lysogenic for recombinant qsr' phage derived from the nmpC (p+) strain contained a new protein identical in electrophoretic mobility to the NmpC porin and to the Lc porin encoded by phage PA-2. Lysogens of qsr' recombinants from the nmpC+ strain and lysogens of lambda p4, which carries the qsr' region, did not produce this protein. When observed by electron microscopy, the DNA acquired from the qsr' prophage showed homology with the region of the DNA molecule of phage PA-2 which contains the lc gene. Relative to that of the recombinant from the nmpC (p+) mutant, the DNA molecule of the recombinant from the nmpC+ parent contained an insertion near the lc gene. These results were supported by blot hybridization analysis of the E. coli chromosome with probes derived from the lc gene of phage PA-2. A sequence homologous to the lc gene was found at the nmpC locus, and the parental strains contained an insertion, tentatively identified as IS5B, located near the 3' end of the porin coding sequence. We conclude that the structural gene for the NmpC porin protein is located within the defective qsr' prophage at 12.5 min on the E. coli K-12 map and that this gene can be activated by loss of an insertion element. PMID- 2984174 TI - Regulatory region of the heat shock-inducible capR (lon) gene: DNA and protein sequences. AB - The CapR protein is an ATP hydrolysis-dependent protease as well as a DNA stimulated ATPase and a nucleic acid-binding protein. The sequences of the 5' end of the capR (lon) gene DNA and N-terminal end of the CapR protein were determined. The sequence of DNA that specifies the N-terminal portion of the CapR protein was identified by comparing the amino acid sequence of the CapR protein with the sequence predicted from the DNA. The DNA and protein sequences established that the mature protein is not processed from a precursor form. No sequence corresponding to an SOS box was found in the 5' sequence of DNA. There were sequences that corresponded to a putative -35 and -10 region for RNA polymerase binding. The capR (lon) gene was recently identified as one of 17 heat shock genes in Escherichia coli that are positively regulated by the product of the htpR gene. A comparison of the 5' DNA region of the capR gene with that of several other heat shock genes revealed possible consensus sequences. PMID- 2984175 TI - Cloning of the Escherichia coli recJ chromosomal region and identification of its encoded proteins. AB - A 9.6-kilobase BamHI-SalI fragment carrying recJ+ was cloned into vector pBR322. Deletion and transposon mutagenesis were used to map the recJ gene on this fragment. The maxicell protein-labeling technique was used to correlate a functional recJ gene with the presence of a polypeptide of 53,000 apparent molecular weight. Two additional genes, one encoding two proteins of 26,000 and 25,000 Mr and the other encoding a 31,000-Mr protein, were mapped on a 3.7 kilobase HindIII-SalI subfragment with recJ. Functions for these adjacent genes are not known; however, insertion mutations in these genes lessen the expression of the putative recJ protein detected in maxicells. A 9.6-kilobase BamHI-SalI fragment carrying the temperature-sensitive mutation recJ147 was also cloned and used for complementation studies to identify other recJ mutations. PMID- 2984177 TI - Vector insertion mutagenesis of Rhizobium sp. strain ORS571: direct cloning of mutagenized DNA sequences. AB - When the limited-host-range plasmid pVP2021 carrying Tn5 was mobilized into Rhizobium sp. strain ORS571 and stable acquisition of Tn5 was selected, ORS571 plasmid-genome cointegrates were exclusively obtained; direct Tn5 transposition was never observed. In every case, genomic cointegrates exhibited an additional (third) IS50 element that bordered VP2021 DNA sequences but maintained a single Tn5 element. Genomic cointegrates containing IS50 triplications were stable; neither phenotypic reversion nor resolution was detectable. Auxotrophic mutant strains (vector insertion mutants) were identified at expected frequencies among derivatives carrying ostensibly random genomic pVP2021 insertions; N2 fixation (Nif)-defective vector insertion mutants were observed among these derivatives at a frequency of 10(-3). The presence of integrated pVP2021 in ORS571 nif::VP2021 mutant genomes enabled VP2021 to constitute an endogenous cloning vector. After EcoRI or KpnI digestions, genomic nif::pVP2021 DNA sequences contiguous with integrated pVP2021 were directly cloned as new replicons without addition of an exogenous vector. Recombinant plasmids derived from two such nif::pVP2021 mutants hybridized to previously analyzed ORS571 Nif DNA sequences. Recombinant plasmid DNA and ORS571 Nif region DNA were found to be colinear; pVP2021 insertions could be accurately mapped. pVP2021 insertion-mutagenesis thus allows the direct cloning of ORS571 gene sequences for which mutant phenotypes can be selected or screened. PMID- 2984176 TI - Composite IS1 elements encoding hydroxamate-mediated iron uptake in FIme plasmids from epidemic Salmonella spp. AB - Eleven FIme plasmids representative of those identified in epidemic strains of Salmonella wien and Salmonella typhimurium isolated in North Africa, Europe, and the Middle East have been examined for the presence of determinants of toxigenicity, adherence, and iron-sequestering mechanisms. Chemical and genetic data indicated that all plasmids code for a hydroxamate-mediated iron assimilation system. Detailed analysis of derivative plasmids and cloned fragments of FIme plasmid pZM61 demonstrated that the general genetic and structural organization of the DNA region containing the genes for hydroxamate biosynthesis and cloacin DF13 receptor was virtually identical to that described for the aerobactin-mediated iron uptake system of pColV-K30. This DNA region is part of a composite element that is 16.7 kilobases long and carries its IS1 modules as inverted repeats. A very similar element is present in either orientation in all nine FIme plasmids analyzed. PMID- 2984178 TI - Isolation and characterization of transposon Tn5-induced symbiotic mutants of Rhizobium loti. AB - Rhizobium loti NZP2037 and NZP2213, each cured of its single large indigenous plasmid, formed effective nodules on Lotus spp., suggesting that the symbiotic genes are carried on the chromosome of these strains. By using pSUP1011 as a vector for introducing transposon Tn5 into R. loti NZP2037, symbiotic mutants blocked in hair curling (Hac), nodule initiation (Noi), bacterial release (Bar), and nitrogen fixation (Nif/Cof) on Lotus pedunculatus were isolated. Cosmids complementing the Hac, Noi, and Bar mutants were isolated from a pLAFR1 gene library of NZP2037 DNA by in planta complementation and found to contain EcoRI fragments of identical sizes to those into which Tn5 had inserted in the mutants. The cosmids that complemented the mutants of these phenotypic classes did not share common fragments, nor did cosmids that complemented four mutants within the Noi class, suggesting that these symbiotically important regions are not tightly linked on the R. loti chromosome. PMID- 2984180 TI - Identification of a bacterial sensing protein and effects of its elevated expression. AB - The Escherichia coli flaA gene product (also called cheC) plays a crucial role in switching flagellar rotational direction during chemotactic responses. Wild-type and mutant alleles have been cloned onto plasmid vectors, and the gene product has been identified as a 37,000-dalton protein. The flaA product appeared as a soluble protein in the cytoplasm when overproduced in minicells and maxicells. The protein could not be detected in flagellar basal structures purified from a wild-type strain. To assess the effects of altered flaA expression, the gene was fused to a synthetic tac promoter that could be regulated by the addition of an inducer. Overproduction resulted in strong counterclockwise flagellar rotational bias and partial paralysis of flagellar motors. These results suggest that the flaA protein provides the interface between the flagellar machinery and the chemotaxis signaling system in a motor structure external to the basal body. PMID- 2984179 TI - Energy recycling by lactate efflux in growing and nongrowing cells of Streptococcus cremoris. AB - Streptococcus cremoris was grown in pH-regulated batch and continuous cultures with lactose as the energy source. During growth the magnitude and composition of the electrochemical proton gradient and the lactate concentration gradient were determined. The upper limit of the number of protons translocated with a lactate molecule during lactate excretion (the proton-lactate stoichiometry) was calculated from the magnitudes of the membrane potential, the transmembrane pH difference, and the lactate concentration gradient. In cells growing in continuous culture, a low lactate concentration gradient (an internal lactate concentration of 35 to 45 mM at an external lactate concentration of 25 mM) existed. The cell yield (Ymax lactose) increased with increasing growth pH. In batch culture at pH 6.34, a considerable lactate gradient (more than 60 mV) was present during the early stages of growth. As growth continued, the electrochemical proton gradient did not change significantly (from -100 to -110 mV), but the lactate gradient decreased gradually. The H+-lactate stoichiometry of the excretion process decreased from 1.5 to about 0.9. In nongrowing cells, the magnitude and composition of the electrochemical proton gradient was dependent on the external pH but not on the external lactate concentration (up to 50 mM). The magnitude of the lactate gradient was independent of the external pH but decreased greatly with increasing external lactate concentrations. At very low lactate concentrations, a lactate gradient of 100 mV existed, which decreased to about 40 mV at 50 mM external lactate. As a consequence, the proton-lactate stoichiometry decreased with increasing external concentrations of protons and lactate at pH 7.0 from 1 mM lactate to 1.1 at 50 mM lactate and at pH 5.5 from 1.4 at l mM lactate to 0.7 at 50 mM lactate. The data presented in this paper suggest that a decrease in external pH and an increase in external lactate concentration both result in lower proton-lactate stoichiometry values and therefore in a decrease of the generation of metabolic energy by the end product efflux process. PMID- 2984181 TI - Construction and application of a promoter-probe plasmid that allows chromogenic identification in Streptomyces lividans. AB - We cloned a Streptomyces coelicolor A3(2) DNA fragment which directed synthesis of a brown pigment, presumably a shunt product in the actinorhodin biosynthetic pathway, on the plasmid vector pIJ41 in Streptomyces lividans. The pigment production was observed only when the DNA fragment was inserted downstream from a functional promoter sequence. By subcloning the fragment together with in vitro manipulation, a promoter-probe plasmid vector (pARC1) with a unique BamHI cloning site was constructed that allows chromogenic identification of transcriptional control signals in Streptomyces lividans based on the expression of the cloned pigment gene(s). The Escherichia coli tac (trp-lac hybrid) promoter, consisting of 92 base pairs and a promoter region including the leader sequence of erythromycin resistance gene (ermC) on staphylococcal plasmid pE194, when ligated in the correct orientation in the BamHI site of pARC1, promoted expression of the cloned pigment gene(s) in Streptomyces lividans, whereas the Saccharomyces cerevisiae GAL7 promoter did not. In the case of the ermC, induction of the pigment production by the addition of either erythromycin or lincomycin, but not virginiamycin, was observed. The system was also shown to be useful and convenient in isolating transcriptional control signals of Streptomyces chromosomal DNA and estimating their activities. PMID- 2984182 TI - Manipulation of intracellular magnesium content in polymyxin B nonapeptide sensitized Escherichia coli by ionophore A23187. AB - Escherichia coli B cells were sensitized to ionophore A23187 by polymyxin B nonapeptide, and the induced magnesium and potassium ion fluxes were studied. Combined ionophore treatment permeabilized the cytoplasmic membrane of E. coli in an ion-specific manner and allowed the manipulation of intracellular Mg2+ content from the outside. A23187-induced Mg2+ efflux or influx was dependent on the free Mg2+ concentration gradient between the outside and inside of the cytoplasmic membrane and on the pH gradient. Most of the intracellular Mg2+ was bound, whereas only 1 to 2 mM was free in solution in the cellular sap. PMID- 2984183 TI - Genetic locus (stmF) associated with cyclic GMP phosphodiesterase activity in Dictyostelium discoideum maps in linkage group II. AB - Previous attempts to map the stmF locus in Dictyostelium discoideum, by using only clone morphology as a marker, have led to equivocal results. Since strains carrying mutations at the stmF locus possess very low cyclic GMP phosphodiesterase activity, we have remapped this locus using both morphological and biochemical markers. Our results indicate that mutations producing a stable "streamer" phenotype and reduced cyclic GMP phosphodiesterase activity are located in linkage group II, probably centromere distal to acrA. PMID- 2984185 TI - Genetic and physical organization of the cloned gyrA and gyrB genes of Bacillus subtilis. AB - An 8-kilobase fragment already known to contain the gyrA gene of Bacillus subtilis was shown to encode the gyrB gene as well. Plasmids containing this fragment can rescue both B. subtilis gyrA and gyrB mutants and complement Escherichia coli gyrA mutants. Deletion analysis has indicated the gene locations on the cloned fragment. Under low-stringency conditions the cloned E. coli gyrA and gyrB genes each hybridized to the appropriate subfragments, confirming the assignment of the gene locations on the cloned DNA. In E. coli maxicells, proteins of 67,000 (gyrA) and 77,000 (gyrB) Mr were synthesized. Analysis of proteins encoded by various subfragments indicated the direction of transcription. Although the gyrA and gyrB genes are located adjacent to each other on the chromosome, they may be transcribed independently since expression of gyrA protein is not dependent upon the gyrB gene in maxicells. PMID- 2984184 TI - Dimeric porin from Paracoccus denitrificans. AB - Paracoccus denitrificans was shown to contain a 33,000-dalton porin, which produced pores of large (1.6 to 1.8 nm) diameter. Cross-linking studies showed that the porin existed as dimers in the outer membrane. PMID- 2984186 TI - Magnetic circular dichroism studies on the heme and tryptophan components of cytochrome c peroxidase. AB - We have measured the magnetic circular dichroism of cytochrome c peroxidase and some of its derivatives from 250-350 nm. Comparison of the changes observed on conversion to the catalytic intermediate (cytochrome c peroxidase-I) with spectra obtained from horseradish peroxidase and its derivatives and model compounds of protoheme leads us to the conclusion that the observed changes in the magnetic circular dichroism spectra reflect conversion of the heme to the ferryl state. No evidence was found for modification of tryptophan in cytochrome c peroxidase-I. PMID- 2984187 TI - The amino acid sequence of an active site peptide from the H,K-ATPase of gastric mucosa. AB - The gastric H,K-ATPase is an active transport protein that is responsible for the maintenance of a large pH gradient across the secretory canaliculus of the mammalian parietal cell. Acid secretion across these epithelial cell membranes is coupled to the potassium-stimulated hydrolysis of ATP catalyzed by H,K-ATPase, but the mechanism of coupling between ion transport and ATP hydrolysis is unknown. In order to investigate the enzymatic mechanism of this coupling, a peptide derived from the ATP binding site of H,K-ATPase has been purified and its amino acid sequence has been determined. The peptide was identified by the incorporation of a fluorescent probe, fluorescein 5'-isothiocyanate (FITC), into the active site before trypsin digestion of the protein. The labeling of the enzyme by FITC was associated with the irreversible inhibition of enzymatic activity, and both the labeling of the tryptic peptide and inhibition of activity were prevented when the reaction was performed in the presence of ATP. At 100% inhibition of activity, 3.5 +/- 1.6 nmol of FITC were incorporated per mg of protein. The amino acid sequence of the active site peptide is His-Val-Leu-Val Met-Lys-Gly-Ala-Pro-Glu-Gln-Leu-Ser-Ile-Arg, and FITC reacts with the lysine. This sequence is very similar to sequences of fluorescein-labeled peptides from the ATP binding sites of Na,K-ATPase and Ca2+-ATPase, and suggests that the active site structures of these ion transport ATPases are similar. PMID- 2984188 TI - Viral Harvey ras p21 expressed in Escherichia coli purifies as a binary one-to one complex with GDP. AB - The ras oncogene of Harvey murine sarcoma virus encodes a 21,000-dalton protein, p21, which mediates transformation. Viral Harvey ras p21, cloned into Escherichia coli HB101 lambda/pRAS1, has been purified to homogeneity by a three-step procedure. The purified E. coli p21 can be bound by a monoclonal antibody to viral Harvey ras p21 and has an amino acid composition consistent with that predicted from its DNA sequence. Purified E. coli p21 has been shown by HPLC analysis on an ion-exchange column to contain near stoichiometric amounts of GDP. This noncovalently associated GDP is seen in the UV absorption spectrum of the purified protein. The noncovalently associated GDP acts as a competitive inhibitor of the interaction of added guanine nucleotides with p21. PMID- 2984189 TI - Enzymatic basis for the structural changes of asparagine-linked sugar chains of membrane glycoproteins of baby hamster kidney cells induced by polyoma transformation. AB - Previous studies indicated that enrichment of the GlcNAc beta 1----6Man alpha 1-- - group with concomitant decrease of the GlcNAc beta 1----4Man alpha 1---- group occurs in the complex-type asparagine-linked sugar chains of the membrane glycoproteins of baby hamster kidney cells transformed by polyoma virus. The enzymatic basis of the chemical change is reported in this paper. By using oligosaccharides isolated from the urine of patients with a variety of exoglycosidase deficiencies, beta-N-acetylglucosaminyltransferases in the cell homogenate were successfully assayed separately. Both baby hamster kidney cells and their polyoma transformants contain beta-N-acetylglucosaminyltransferases I, II, IV, V, and VI, but not beta-N-acetylglucosaminyltransferase III. The beta-N acetylglucosamine residue added by each beta-N-acetylglucosaminyltransferase (GnT) is shown below. (formula see text) Comparative studies of the specific activities of the five beta-N-acetylglucosaminyltransferases in the two cell lines revealed that the value of beta-N-acetylglucosaminyltransferase V in the polyoma transformant was twice of that in the normal cells, while those of the other four transferases in the two cell lines were not significantly different. Therefore the increase in beta-N-acetylglucosaminyltransferase V may be the direct cause of the changes found in the sugar chains of surface glycoproteins in baby hamster kidney cells transformed by polyoma virus. PMID- 2984190 TI - Acetate biosynthesis by acetogenic bacteria. Evidence that carbon monoxide dehydrogenase is the condensing enzyme that catalyzes the final steps of the synthesis. AB - The purified carbon monoxide dehydrogenase from Clostridium thermoaceticum is the only protein required to catalyze an exchange reaction between carbon monoxide and the carbonyl group of acetyl-CoA. This exchange requires that the CO dehydrogenase bind the methyl, the carbonyl, and the CoA groups of acetyl-CoA, then equilibrate the carbonyl with CO in the solution and re-form acetyl-CoA. CoA is not necessary for the exchange and, in fact, inhibits the reaction. These studies support the view that CO dehydrogenase is the condensing enzyme that forms acetyl-CoA from its component parts. Carbon dioxide also exchanges with the C-1 of acetyl-CoA, but at a much lower rate than does CO. At 50 degrees C and pH 5.3, the optimal pH, the turnover number is 70 mol of CO exchanged per min/mol of enzyme. Low potential electron carriers are stimulatory. The Km app for stimulation by ferredoxin is 50-fold less than the value for flavodoxin. Neither ATP or Pi stimulate the exchange. The EPR spectrum of the CO-reacted enzyme is markedly changed by binding of CoA or acetyl-CoA. Arginine residues of the CO dehydrogenase appear to be involved in the active site, possibly by binding acetyl-CoA. Mersalyl acid, methyl iodide, 5,5-dithiobis-(2-nitrobenzoate), and sodium dithionite inhibit the exchange reaction. A scheme is presented to account for the role of CO dehydrogenase in the exchange reaction and in the synthesis of acetate. PMID- 2984191 TI - The effect of organic solvents on the activation and the activity of spinach chloroplast fructose-1,6-bisphosphatase. AB - A two-stage assay was used to study the effect of organic solvents on the activation of and the catalysis by chloroplast fructose-1,6-bisphosphatase. Irrespective of chemical structure, all the organic solvents tested had a dual effect on the enzyme. In the activation they stimulated and inhibited at low and high concentrations, respectively, in a process that required dithiothreitol, fructose 1,6-bisphosphate, and Ca2+. Conversely, organic solvents inhibited catalysis. The enhancement in fructose-1,6-bisphosphatase activity did not arise from a change in the molecular weight of the enzyme and correlated positively with the hydrophobic character of the organic solvent. In the presence of 2 propanol, all the activation constants for modulators (fructose 1,6-bisphosphate, a2+, thioredoxin-f) were lower than in a strictly aqueous medium. Monothiols were also functional in the activation of chloroplast fructose-1,6-bisphosphatase, although they were less effective than dithiols. Sulfhydryl compounds decreased the concentration of fructose 1,6-bisphosphate required for the activation of the enzyme, and 2-propanol lowered this requirement further. Arrhenius plots were nonlinear for the enzyme activation and linear for the hydrolytic step. The anomalous temperature dependence of the chloroplast fructose-1,6-bisphosphatase activation was indicative of a cooperative process. The data obtained in this study indicate that the concerted activation of chloroplast fructose-1,6 bisphosphatase is favored in a medium less polar than water. PMID- 2984193 TI - Spin trapping of the azidyl radical in azide/catalase/H2O2 and various azide/peroxidase/H2O2 peroxidizing systems. AB - The azidyl radical is formed during the oxidation of sodium azide by the catalase/hydrogen peroxide system, as detected by the ESR spin-trapping technique. The oxidation of azide by horseradish peroxidase, chloroperoxidase, lactoperoxidase, and myeloperoxidase also forms azidyl radical. It is suggested that the evolution of nitrogen gas and nitrogen oxides reported in the azide/catalase/hydrogen peroxide system results from reactions of the azidyl radical. The azide/horseradish peroxidase/hydrogen peroxide system consumes oxygen, and this oxygen uptake is inhibited by the spin trap 5,5-dimethyl-1 pyrroline-N-oxide, presumably due to the competition with oxygen for the azidyl radical. Although azide is used routinely as an inhibitor of myeloperoxidase and catalase, some consideration should be given to the biochemical consequences of the formation of the highly reactive azidyl radical by the peroxidase activity of these enzymes. PMID- 2984192 TI - Activation of the human neutrophil NADPH oxidase results in coupling of electron carrier function between ubiquinone-10 and cytochrome b559. AB - The enzymatic activity underlying the respiratory burst in human neutrophils was examined in a subcellular fraction with high specific activity and shown to be a membrane-associated complex of a flavoprotein, ubiquinone-10, and cytochrome b559 in an approximate 1.3:1:2 molar ratio. Study of the redox poise of these electron carriers indicated that electron flow in the intact complex from unstimulated cells proceeded: NADPH----E-FAD----ubiquinone-10. Similar studies on the complex prepared from stimulated neutrophils indicated that electron flow proceeded: NADPH----E-FAD----ubiquinone-10----cytochrome b559----oxygen. The active enzyme complex was inhibited by p-chloromercuribenzoate. Inhibition persisted after removal of excess inhibitor, was reversed by dithiothreitol, and could be blocked by prior addition of substrate (NADPH). Inhibition of the active oxidase complex by p-chloromercuribenzoate also inhibited electron flow from NADPH to all purported electron carriers in the chain (i.e. E-FAD, ubiquinone-10, and cytochrome b559). We conclude that activation of the oxidase enzyme complex in the intact neutrophil resulted in linkage of electron carrier function between endogenous ubiquinone-10 and cytochrome b559 and was without demonstrable effect on proximal electron flow. The p-chloromercuribenzoate sensitive site(s) proximal to the initial electron acceptor (E-FAD) did not appear to be altered by the cellular activation process. PMID- 2984194 TI - sn-1,2-Diacylglycerol kinase of Escherichia coli. Purification, reconstitution, and partial amino- and carboxyl-terminal analysis. AB - The sn-1,2-diacylglycerol kinase structural gene from Escherichia coli was demonstrated to be the dgkA locus previously sequenced (Lightner, V. A., Bell, R. M., and Modrich, P. (1983) J. Biol. Chem. 258, 10856-10861). The dgkA gene product was shown by maxicell analysis to be an Mr = 14,000 membrane-bound protein. When dgkA was placed on a hybrid plasmid under control of the lambda pL promoter, a 100-fold overproduction of diacylglycerol kinase activity was obtained. Diacylglycerol kinase was solubilized from membranes with 2 propanol/heptane/trifluoroacetic acid and purified to near homogeneity by high performance liquid chromatography. Activity was reconstituted in a mixed micellar assay containing beta-octylglucoside, cardiolipin, and sn-1,2-dioleoylglycerol. Amino acid analysis, partial NH2-terminal analysis and COOH-terminal analysis permitted alignment of the polypeptide on the sequenced gene. The data establish that dgkA is the structural gene for the diacylglycerol kinase and establish the primary structure of the enzyme of 122 residues, 13,245 daltons. Secondary structure analysis predicted a protein conformation consisting of three transmembrane alpha-helical segments, an amphipathic helix, and an alpha-helix. Taken together, the predicted helical segments comprise more than 75% of the polypeptide. PMID- 2984196 TI - Investigation of the selectivity of hydrogen abstraction in the nonenzymatic formation of hydroxyeicosatetraenoic acids and leukotrienes by autoxidation. AB - The biosynthetic conversions of arachidonic acid to hydroperoxyeicosatetraenoic acids (HPETEs) and the further conversion of leukotriene epoxides are accompanied by stereoselective hydrogen abstraction from the reaction substrate. Furthermore, this hydrogen removal has always been found to occur in fixed stereochemical relationship to carbon-oxygen chiral center(s) in the substrate or product. We have used stereospecifically labeled 10-3H-substrates with 14C internal standard to investigate whether the same relationships bear in HPETE and leukotriene formation during autoxidation. After autoxidation of labeled arachidonate, both the 8(R)- and 8(S)-HPETE enantiomers (resolved as diastereomer derivatives) and the 12(RS)-HPETE were observed to retain 41-47% 3H relative to the starting material. In autoxidative formation of leukotrienes from labeled 15(S)-HPETE the four main leukotrienes, including two derived from 14,15-leukotriene A4 hydrolysis, were observed to have retained an average of 45% 3H. Primary and secondary isotope effects were found to accompany these reactions. The results prove that stereorandom hydrogen abstraction occurs in autoxidation and that the hydrogen loss bears no stereochemical relationship to chiral oxygen center(s) in the HPETE product, (8(R) or 8(S], or the 15(S)-hydroperoxy substrate. We conclude that the chiral features of the biosynthetic reactions are a reflection of enzymatic control of stereochemistry. Nonetheless, the findings of primary and secondary isotope effects in autoxidation which are similar to those observed in the analogous biosynthetic reactions suggests that, except for stereochemical control, the autoxidative and enzymatic reactions may be mechanistically similar. PMID- 2984195 TI - Demonstration of different metal ion-induced calcineurin conformations using a monoclonal antibody. AB - It has been suggested that calcineurin, a calmodulin-stimulated phosphatase, may exist in different metal ion-dependent conformational states (Pallen, C.J., and Wang, J. H. (1984) J. Biol. Chem. 259, 6134-6141). Evidence in favor of this hypothesis comes from studies involving a monoclonal antibody, VA1, which is specific for the small (beta) subunit of calcineurin. This antibody inhibits Ni2+ stimulated but not Mn2+-stimulated phosphatase activity against p-nitrophenyl phosphate and phosphorylase kinase. Inhibition is not due to competition of the antibody with substrate or to interference with metal ion binding to the enzyme. Complex formation between the antibody and calcineurin can be demonstrated either in the presence of Mn2+ or Ni2+ or in the absence of metal ion activators. These results indicate that the active conformational states of calcineurin are metal ion dependent, that the monoclonal antibody VA1 affects the Ni2+-induced conformational change of the enzyme, and that the beta subunit of calcineurin plays a critical role in the expression of Ni2+-stimulated phosphatase activity. PMID- 2984197 TI - Investigation of the chemical conversion of hydroperoxyeicosatetraenoate to leukotriene epoxide using stereospecifically labeled arachidonic acid. Comparison with the enzymatic reaction. AB - A series of stereospecifically labeled polyunsaturated fatty acids were prepared by biosynthesis from [8-DR-3H]- and [8-LS-3H]stearic acids. The labeled stearic acids were synthesized by a novel scheme employing readily available alkyne and aldehyde starting materials. The stereochemical purity of the prochiral tritium labels was judged to be greater than 99%, as determined by analysis of the octadec-1-yn-8(R)- and 8(S)-ol intermediates in the synthesis. Previously, the labeled arachidonic acids were used to investigate the stereoselectivity of hydrogen abstraction in the biosynthesis of leukotriene epoxides. We have now investigated the selectivity of hydrogen abstraction in a chemical synthesis of 14,15-leukotriene (LT) A4 from mixtures of [3-14C]- and either [10-DR-3H]- or [10 LS-3H]15(S)-HPETE methyl esters. Reaction with either chirally labeled precursor led to 70-95% retention of 3H relative to 14C in the 14,15-LTA4 and 10-Z-14,15 LTA4 products after purification by high performance liquid chromatography. The 15-dienone obtained from this reaction was consistently enriched in 3H relative to 14C after isolation and purification. Evidence was obtained to indicate that the majority of the 3H in the products was retained in its original location and configuration. These results indicate that the biomimetic chemical reaction is stereo-random with respect to hydrogen loss from carbon 10 and that, in contrast to the reaction as it occurs in leukocytes and platelets, in the chemical model the reaction begins by decomposition of the hydroperoxide group, with hydrogen loss from carbon 10 occurring as a late or final step. PMID- 2984198 TI - DNA sequence analysis of the dye gene of Escherichia coli reveals amino acid homology between the dye and OmpR proteins. AB - Mutation of the dye gene of Escherichia coli results in sensitivity of dyes, envelope protein changes, loss of expression of alkaline phosphatase, and reduced transcription of sex factor F genes. We have determined the DNA sequence of a 1.4 kilobase pair fragment encompassing the dye gene. The coding sequence of dye was identified as an open reading frame coding for a protein of Mr 27,346. A sequence of 54 residues at the amino terminus was extremely acidic, with 12 aspartic plus glutamic acid residues and only 2 lysine plus arginine residues. A sequence of 19 adjacent residues near the center of the protein was identical, except for one mismatch, with a sequence in the OmpR protein, involved in controlling the amounts of the major outer membrane proteins OmpF and OmpC at the level of transcription. 28% of the Dye protein was homologous with OmpR. The positions of dye and ompR on the genetic map were indicative of a gene duplication. It seems likely, therefore, that the Dye and OmpR proteins are related, and Dye may thus be involved in the osmoregulation of envelope protein genes as well as being required for sex factor gene expression. The Dye protein itself, like OmpR, was shown not to be an envelope protein. A second open reading frame on the other DNA strand may use the same transcription termination site as dye. PMID- 2984199 TI - Phosphorylase phosphatase regulatory subunit. "Western" blotting with immunoglobulins against inhibitor-2 reveals a protein of Mr = 60,000. AB - Rabbit muscle phosphorylase phosphatase has been isolated in different laboratories as an inactive complex of Mr = 70,000, composed of separate catalytic (Mr = 38,000) and regulatory (Mr = 31,000) proteins. The regulatory protein is identical to one of two heatstable inhibitors called inhibitor-2 (I2). Antiserum raised in sheep against I2 by repeated immunization potently blocked inhibitory activity, whereas preimmune serum did not. Immunoglobulins which blocked inhibitory activity were purified by affinity chromatography with I2 as the immobilized ligand. Using a "Western" immunoblotting procedure, as little as 1-5 ng of pure I2, obtained by electroelution of the Mr = 31,000 band of I2 from a polyacrylamide gel segment, were detected. Immunoblotting of the immunogen revealed only a band at Mr = 31,000, indicating the absence of contaminating antigenic proteins. When extracts of skeletal muscle and other rabbit tissues were denatured directly in dodecyl sulfate for immunoblotting the most intensely stained band was present at Mr = 60,000, rather than at Mr = 31,000 as expected. A small amount of I2 and other bands were detected, in particular at Mr = 36,000 and 25,000. Subsequent to heat treatment of the tissue extracts, there was an enrichment of I2 content relative to the Mr = 60,000 band. The results indicate the existence of a Mr = 60,000 protein related to I2. Activation of phosphorylase phosphatase in a muscle extract by treatment with Co2+ plus trypsin exactly coincided with digestion of the Mr = 60,000 immunoreactive protein. Available data indicate that this protein may function as a regulatory subunit of phosphorylase phosphatase. PMID- 2984200 TI - Phosphorylase phosphatase catalytic subunit. Evidence that the Mr = 33,000 enzyme fragment is derived from a native protein of Mr = 70,000. AB - An active form of phosphorylase phosphatase of Mr = 33,000, referred to as the catalytic subunit for over a decade, was purified to near-homogeneity from rabbit skeletal muscle. Repeated immunization of a sheep produced immunoglobulins that blocked the activity of the phosphatase. These immunoglobulins were affinity purified on columns of immobilized phosphorylase phosphatase and used as macromolecular probes in a "Western" immunoblotting procedure with peroxidase conjugated rabbit anti-sheep immunoglobulins. Only one protein, of Mr = 33,000, was stained in samples of the immunogen, attesting to the specificity of the probes. However, the Mr = 33,000 phosphatase protein was not detected in muscle extracts or in partially purified preparations. Instead, a single protein of Mr = 70,000 was detected. Limited proteolysis, in particular by Staphylococcus aureus V8 protease and thermolysin, converted the immunoreactive protein from Mr = 70,000 to Mr = 33,000. Coagulation of the phosphatase preparation with 80% ethanol at room temperature rendered the Mr = 70,000 protein insoluble, but allowed extraction of the Mr = 33,000 protein from the precipitate. Thus, we conclude that the immunoreactive protein of Mr = 70,000 is the "catalytic subunit" of phosphorylase phosphatase with a catalytic domain of Mr = 33,000. Previous purification schemes have yielded only the fragment of Mr = 33,000 due to its relative resistance to proteolysis and coagulation. Gel filtration chromatography of the "native" form of phosphorylase phosphatase showed Mr approximately 230,000. Both the Mr = 70,000 catalytic subunit and a Mr = 60,000 protein related to inhibitor-2 were detected by immunoblotting in the same fractions that exhibited activity after treatment with Co2+ and trypsin. Only the Mr = 60,000 protein was degraded during this activation process. We propose that the native phosphorylase phosphatase is an elongated structure with two-fold symmetry, containing one catalytic subunit of Mr = 70,000 and one regulatory subunit of Mr = 60,000. PMID- 2984201 TI - Mechanism of interferon action: inhibition of vesicular stomatitis virus replication in human amnion U cells by cloned human gamma-interferon. I. Effect on early and late stages of the viral multiplication cycle. AB - The molecular basis of the inhibition of vesicular stomatitis virus (VSV) replication by pure recombinant gamma-interferon (IFN-gamma) in human amnion U cells was examined. A saturating concentration of IFN-gamma induced, at maximum, about a two log10 reduction in infectious VSV yield. The kinetics of induction of the antiviral activity by IFN-gamma were first order over the period of about 6 18 h, following a lag of about 3 h, after treatment with a saturating concentration of IFN-gamma. The relationship of the inhibition in VSV infectivity to the early and late events of the VSV multiplication cycle was investigated. IFN-gamma treatment had no detectable effect on the adsorption and penetration of VSV virions or on their uncoating to yield viral nucleocapsids. The polypeptides of adsorbed or uncoated VSV particles were neither preferentially degraded nor detectably altered in IFN-gamma-treated U cells, as compared to untreated U cells. Progeny virions isolated from IFN-gamma-treated U cells, although greatly reduced in number, were found to be equally as infectious as those isolated from untreated U cells. Progeny virions from IFN-gamma-treated cells also possessed the same composition of viral proteins as was observed for virions from untreated cells. These results suggest that conditions of IFN-gamma treatment sufficient to reduce the yield of infectious VSV progeny 100-fold do not detectably affect either the early or the late stages of the VSV multiplication cycle. PMID- 2984202 TI - Mechanism of interferon action: inhibition of vesicular stomatitis virus replication in human amnion U cells by cloned human gamma-interferon. II. Effect on viral macromolecular synthesis. AB - The effects of recombinant human gamma-interferon (IFN-gamma) on vesicular stomatitis virus (VSV) macromolecular synthesis in human amnion U cells were examined. Saturating concentrations of IFN-gamma caused only a 3 to 5-fold reduction of viral protein synthesis in wild-type VSV-infected cells, an extent insufficient to account for the 100-fold inhibition of viral infectivity. By use of the VSV mutant tsG41, which is competent in RNA transcription but defective in RNA replication at 40 degrees C, it was shown that the apparent IFN-induced inhibition of viral protein synthesis was likely due to a reduction in the synthesis of primary transcripts in IFN-gamma-treated U cells. Dot blot hybridization analysis revealed that saturating concentrations of IFN-gamma reduced both primary (measured with mutant tsG41-infected U cells) and total (measured with wild-type-infected U cells) viral RNA synthesis by about 4-fold, an extent of inhibition comparable to the observed reduction in viral protein synthesis. Analysis of RNA, fractionated by agarose gel electrophoresis after denaturation with glyoxal, with cDNA probes to individual VSV mRNAs did not reveal any detectable difference in the structural integrity of VSV mRNA isolated from IFN-gamma treated as compared to untreated U cells. These results suggest that IFN-gamma treatment causes a small reduction in the efficiency of transcript formation catalyzed by input parental virions. However, the results also indicate that the principal cause of the IFN-gamma-induced inhibition of VSV replication in U cells is the alteration of a step in replication other than viral macromolecular synthesis. This implies that the molecular mechanism of viral inhibition by IFN-gamma is fundamentally different from that of IFN-alpha in human amnion U cells. PMID- 2984203 TI - Glyceraldehyde-3-phosphate dehydrogenase from Drosophila melanogaster. Identification of two isozymic forms encoded by separate genes. AB - The enzyme glyceraldehyde-3-phosphate dehydrogenase from Drosophila melanogaster has been purified, and these preparations contain two subunits forms which have molecular weights of 37,000 and 35,500, respectively. Each subunit is found in crude extracts, and two activity bands are seen in nondenaturing acrylamide gels. Translation of Drosophila poly(A)-containing RNA results in two products which are precipitable with anti-glyceraldehyde-3-phosphate dehydrogenase serum. Two recombinant DNA clones have been isolated from a genomic library of Drosophila DNA. Each of these clones has the ability to hybrid select mRNAs which translate into both subunit forms. These clones have been genetically characterized by in situ hybridization and restriction mapping. One clone hybridizes to region 13F and the other to region 43E of the Drosophila cytogenetic map. Therefore, it appears that the Drosophila melanogaster genome contains two unlinked genes for glyceraldehyde-3-phosphate dehydrogenase; one of them encodes a protein of 37,000 daltons, the other a protein of 35,500 daltons. PMID- 2984204 TI - Structure of the promoter of the rat type II procollagen gene. AB - We have isolated several overlapping genomic clones which contain the 5' terminal portion of the rat pro-alpha 1 (II) chain gene. These clones span about 20 kilobases (kb) of contiguous DNA containing 15 kb of the gene and 5 kb of the 5' flanking sequence. Electron microscopic analysis of mRNA-DNA hybrids by R-looping shows that collectively these clones contain 16 exons which code for approximately one-third of the pro-alpha 1 (II) chain. The sizes of the exons are small, except for the first exon which is relatively large. The nucleotide sequence of the first exon and the 1000 base pairs (bp) preceding it was determined. The first exon contains a 150-bp untranslated segment and an 85-bp sequence coding for the signal peptide and a part of the NH2-terminal propeptide of type II collagen. The segment preceding the transcription initiation site contains the "TATA" box and several G + C-rich stretches, whereas the "CAT" box is not evident between -70 and -120. The hexanucleotide sequence 5'-GGGCGG-3' is found in three different places between -200 and the TATA box. The inverted complement sequence of this hexanucleotide, 5'-CCGCCC-3', is located around both 220 and -450. The hexanucleotide and its inverted sequence have been found previously in the promoter region of the tk gene of herpes virus. These sequences are known to function in a mutually dependent manner as transcription signals for the tk gene; thus, they may play a role in determining the level of transcription of this cartilage gene. The hexanucleotide, 5'-CCGCCC-3', is also found in the 21 base pair repeats of the SV40 promoter and the promoter region of hydroxymethylglutaryl-CoA reductase gene. The sequence 5'-GTGGTTAGA-3' located around -280 is identical to the "core" sequence that has been reported as enhancer element in both viral and cellular genes. These unusual structures may be related to the tissue-specific expression of this gene. PMID- 2984205 TI - The phi 80 and P22 attachment sites. Primary structure and interaction with Escherichia coli integration host factor. AB - Although the lambdoid bacteriophage phi 80 and P22 possess site-specific recombination systems analogous to bacteriophage lambda, they have different attachment (att) site specificities. We have identified and determined the nucleotide sequences of the att sites of phi 80 and P22 and have examined the interaction of these sites with purified Escherichia coli integration host factor (IHF). The sizes of the homologous core regions of the att sites vary greatly: P22 has a 46-base pair core, while phi 80 and lambda have 17- and 15-base pair cores, respectively. The core sequences of the three phage show no significant homology, although dispersed regions of homology in arm sequences indicate that the three phage att sites are related. All three att sites have a high A + T composition, and restriction fragments carrying these sites migrate anomalously upon polyacrylamide gel electrophoresis. IHF binds to a site to the left of the common core in the phi 80 and P22 phage att sites (attP) and to a site to the right of the core in P22 attP and attB (the bacterial att site). In the lambda system, IHF interacts with three regions on attP (designated H1, H2, and H') and none on attB (Craig N., and Nash, H.A. (1984) Cell 39, 707-716). Alignment of the IHF sites of all three phage results in a consensus sequence for IHF binding, Pyr AANNNNTTGATAT. Among the three phage, the number of IHF sites differs; however, the location and orientation of the binding sites in relation to the respective core regions are well conserved. An IHF site analogous to lambda H2 is present in both phi 80 and P22 attP, while a site analogous to lambda H' is present in P22 attP. This conservation suggests that IHF plays a very similar role in the site specific recombination pathways of all three phage, and that the flanking arm sequences are necessary for phi 80 and P22 attP function, as is the case for lambda attP function. These structural similarities presumably reflect a conservation of the mechanism of site-specific recombination for the three phage. PMID- 2984206 TI - Isolation and characterization of the chicken cardiac myosin light chain (L-2A) gene. Evidence for two additional N-terminal amino acids. AB - The contractile proteins of striated muscle are encoded by multigene families and constitute an excellent system to investigate differentiation and developmental control of gene expression. Different forms of myosin light chains are expressed in skeletal muscle as well as in the myocard. To study the gene structure and molecular mechanisms underlying differential gene expression, the structural cardiac myosin light chain 2 (MLC-2A) gene was isolated from a chicken genomic DNA library. Restriction enzyme mapping, electron microscopic analysis, and partial sequencing revealed that the gene coding for the MLC mRNA of 700 nucleotides in length extends over 4.2 kilobases of DNA and is interrupted by 5 introns. Sequence analysis led to the detection of two codons for additional amino acids at the N terminus which were not reported to be present in the mature protein and are presumably removed post-translationally. These two amino acids, methionine and alanine, are coded on two separate exons split by the largest intron of the entire gene. Southern blot analysis of genomic chicken DNA indicates the presence of one MLC-2A gene per haploid chicken genome. PMID- 2984207 TI - Relaxation of supercoiled phosphorothioate DNA by mammalian topoisomerases is inhibited in a base-specific manner. AB - The nucleotide preferences of calf thymus topoisomerases I and II for recognition of supercoiled DNA have been assessed by the relaxation and cleavage of DNA containing base-specific phosphorothioate substitutions in one strand. The type I enzyme is inhibited to varying degrees by all modified DNAs, but most effectively (by approximately 60%) if deoxyguanosine 5'-O-(1-thiomonophosphate) (dGMP alpha S) is incorporated into negatively supercoiled DNA. A DNA in which all internucleotide linkages of one strand are phosphorothionate is relaxed, most probably via the unsubstituted strand. The type II enzyme is inhibited when deoxyadenosine 5'-O-(1-thiomonophosphate) (dAMP alpha S) or deoxyribosylthymine 5'-O-(1-thiomonophosphate) is incorporated into the DNA substrate, and the course of the relaxation reaction changes from a distributive mode to a predominantly processive mode. A fully substituted DNA is very poorly relaxed by the type II enzyme, illustrating the strict commitment of the enzyme to relaxation via double strand cleavage. The sense of supercoiling does not affect the inhibition profile of either enzyme. DNA strand breaks introduced by type II topoisomerase in a normal control DNA or deoxycytidine 5'-O-(1-thiomonophosphate)-substituted DNA on treatment with sodium dodecyl sulfate at low ionic strength are prevented by pretreatment with 0.2 M NaCl. In contrast, breaks in DNA having either dAMP alpha S or all four phosphorothioate nucleotides incorporated in one strand are prevented only with higher NaCl concentrations. Thus indicating activity at the phosphorothioate linkage 5' to dA but not 5' to dC. We conclude that topoisomerase II activity occurs preferentially at sites possessing dAMP or dTMP, and that dGMP is involved in DNA recognition by topoisomerase I. PMID- 2984208 TI - Mercuric and cadmium ions stimulate phosphorylation of band 4.2 protein on human erythrocyte membranes. AB - In this study, we found that Hg2+ and Cd2+ enhanced the phosphorylation of human erythrocyte membranous proteins, especially band 4.2 protein, which was hardly phosphorylated in the absence of the metal ions. p-Chloromercuribenzenesulfonate and p-chloromercuribenzoate had effects similar to those of Hg2+ and Cd2+ on band 4.2 protein phosphorylation, while other metal ions and sulfhydryl agents, such as N-ethylmaleimide, 5,5'-dithiobis-(2-nitrobenzoic acid), or iodoacetate, did not. The Hg2+-stimulated phosphorylation of band 4.2 protein required a millimolar concentration of Mg2+, and it was inhibited by Ca2+ dose-dependently. Phosphoserine was identified from a hydrolysate of the phosphorylated band 4.2 protein by high-voltage electrophoresis. A specific protein inhibitor against cAMP-dependent protein kinase decreased the Hg2+-stimulated phosphorylation of band 4.2 protein. This protein had more binding sites for 203Hg2+ than any other membrane proteins. A spectrin complex from the Hg2+-treated membranes contained the band 4.2 protein, which was not detected in the complex from untreated membranes. Furthermore, protein kinase, which could phosphorylate the band 4.2 protein, was also contained in the cytoskeletal fraction from the Hg2+-treated membranes. These results suggest that Hg2+ may bind certain sulfhydryl groups of band 4.2 and other proteins to make band 4.2 protein susceptible to the endogenous cAMP-dependent protein kinase. PMID- 2984209 TI - Adjuvant chemotherapy for malignant fibrous histiocytoma in the femur and tibia. AB - Between 1974 and 1982, twelve patients with malignant fibrous histiocytoma of the femur and tibia were treated by excision and adjuvant chemotherapy. During the same time-period, eighteen patients were treated by excision alone. The disease free survival rate among the former was 59 per cent (seven of twelve). There was only one survival in the latter group. PMID- 2984210 TI - Pulmonary hyalinizing granulomas. AB - Little has been written about pulmonary hyalinizing granuloma as a cause of nodules in the lungs. In a patient with a past history of carcinoma of the breast, the diagnosis made a significant difference in the prognosis. PMID- 2984211 TI - SOS-type functions in mammalian cells. Enhanced reactivation of UV-irradiated SV 40 in UV-irradiated CV-1 cells. AB - The reactivation of UV-irradiated SV 40 was measured in UV-irradiated CV-1 cells as a function of time between irradiation of the cells and infection. To avoid the possible bias of multiplicity reactivation and/or virus propagation, infection was quantitated in terms of V-antigen-positive nuclei at the end of the first lytic cycle. In irradiated cell cultures enhanced reactivation of SV 40 was observed, which indicates induction of DNA repair enzymes. Maximal reactivation was obtained when the time interval between irradiation and infection was 72 h. The UV-inducible DNA repair enzymes might represent elements of an SOS-type response in mammalian cells. PMID- 2984212 TI - Preservation of high energy phosphates in hypertrophied human myocardium. AB - Hypertrophied hearts are extremely vulnerable to ischemia. In 61 patients with aortic valve disease undergoing surgery we investigated the quality of myocardial protection obtained with three different methods of crystalloid cardioplegia. To exclude the influence of temperature differences the regional myocardial temperatures were continuously measured and adjusted to the same level in all patients. Before and after ischemia and after 10 minutes reperfusion myocardial biopsies were taken and the high energy phosphates and lactic acid determined. In one group St. Thomas cardioplegia was used, in another Bretschneider cardioplegia and in the third the Hamburg method. There were no significant differences between the three groups at the end of ischemia and after 10 minutes reperfusion. After 10 minutes reperfusion the metabolic alterations caused by ischemia were in part reversible. From our results we conclude that each of the cardioplegias used is able to protect a hypertrophied heart adequately against ischemia during cardiac arrest. PMID- 2984213 TI - Endogenous and exogenous domain markers of the rat hepatocyte plasma membrane. AB - We have used a combined biochemical and morphological approach to establish the suitability of certain endogenous and exogenous domain markers for monitoring the separation of rat hepatocyte plasma membrane domains in sucrose density gradients. As endogenous domain markers, we employed two of the integral plasma membrane protein antigens, HA 4 and CE 9, localized to the bile canalicular and sinusoidal/lateral domains, respectively, of the hepatocyte plasma membrane in rat liver tissue (Hubbard, A. L., J. R. Bartles, and L. T. Braiterman, 1985, J. Cell Biol., 100:1115-1125). We used immunoelectron microscopy with a colloidal gold probe to demonstrate that HA 4 and CE 9 retained their domain-specific localizations on isolated hepatocyte plasma membrane sheets. When the plasma membrane sheets were vesiculated by sonication and the resulting vesicles were centrifuged to equilibrium in sucrose density gradients, quantitative immunoblotting revealed that the vesicles containing HA 4 and those containing CE 9 exhibited distinct density profiles. The density profile for the bile canalicular vesicles (marked by HA 4) was characterized by a single peak at a density of 1.10 g/cm3. The density profile for the sinusoidal/lateral vesicles (marked by CE 9) was bimodal, with a peak in the body of the gradient at a density of 1.14 g/cm3 and a smaller amount in the pellet (density greater than or equal to 1.17 g/cm3). We used this sucrose gradient fractionation as a diagnostic procedure to assign domain localizations for several other hepatocyte plasma membrane antigens and enzyme activities. In addition, we used the technique to demonstrate that 125I-wheat germ agglutinin, introduced during isolated liver perfusion at 4 degrees C, can serve as an exogenous domain marker for the sinusoidal domain of the rat hepatocyte plasma membrane. PMID- 2984214 TI - Effect of viral infection on host protein synthesis and mRNA association with the cytoplasmic cytoskeletal structure. AB - We studied the association of several eucaryotic viral and cellular mRNAs with cytoskeletal fractions derived from normal and virus-infected cells. We found that all mRNAs appear to associate with the cytoskeletal structure during protein synthesis, irrespective of their 5' and 3' terminal structures: e.g., poliovirus that lacks a 5' cap structure or reovirus and histone mRNAs that lack a 3' poly A tail associated with the cytoskeletal framework to the same extent as capped, polyadenylated actin mRNA. Cellular (actin) and viral (vesicular stomatitis virus and reovirus) mRNAs were released from the cytoskeletal framework and their translation was inhibited when cells were infected with poliovirus. In contrast, actin mRNA was not released from the cytoskeleton during vesicular stomatitis virus infection although actin synthesis was inhibited. In addition, several other conditions under which protein synthesis is inhibited did not result in the release of mRNAs from the cytoskeletal framework. We conclude that the association of mRNA with the cytoskeletal framework is required but is not sufficient for protein synthesis in eucaryotes. Furthermore, the shut-off of host protein synthesis during poliovirus infection and not vesicular stomatitis virus infection occurs by a unique mechanism that leads to the release of host mRNAs from the cytoskeleton. PMID- 2984216 TI - Phenotypic transformation of normal rat kidney cells in a growth-factor-defined medium: induction by a neuroblastoma-derived transforming growth factor independently of the EGF receptor. AB - Polypeptide growth factor activity in serum can be destroyed by treatment with dithiothreitol. When such growth-factor-inactivated serum is used as a supplement of culture media instead of regular serum, normal rat kidney (NRK) cells become quiescent unless defined polypeptide growth factors like insulin and epidermal growth factor (EGF) are added. On this basis a growth-factor-defined medium has been developed for NRK cells, which permits cell proliferation as rapidly as in media supplemented with serum, even at low cell densities. Moreover, cells can be serially passaged in this medium. NRK cells can be induced to grow in semisolid media when incubated with transforming growth factors. The growth-factor-defined medium permits soft agar growth experiments of NRK cells, without interference from polypeptide growth factors in serum. Using this assay system we have shown that EGF alone is unable to induce any degree of anchorage-independent growth in NRK cells. However, a recently identified transforming growth factor from mouse neuroblastoma cells which does not compete with EGF for receptor binding is able to induce progressively growing colonies of NRK cells in soft agar, even without additional EGF. PMID- 2984215 TI - Muscarinic receptor-induced phosphoinositide hydrolysis at resting cytosolic Ca2+ concentration in PC12 cells. AB - In PC12 cells, cultured in the presence of nerve growth factor to increase their complement of muscarinic receptors, treatment with carbachol induces muscarinic receptor-dependent rises in free cytosolic Ca2+ as well as hydrolysis of membrane phosphoinositides. Experiments were carried out to clarify the relationship between these two receptor-triggered events. In particular, since inositol-1,4,5 trisphosphate (the hydrophilic metabolite produced by the hydrolysis of phosphatidylinositol-4,5-bisphosphate) is believed to mediate intracellularly the release of Ca2+ from nonmitochondrial store(s), it was important to establish whether it can be generated at resting cytoplasmic concentration of Ca2+ (approximately 0.1 microM). Cells incubated in Ca2+-free medium were depleted of their cytoplasmic Ca2+ stores by pretreatment with ionomycin. When these cells were then treated with carbachol, their cytosolic concentration of Ca2+ remained at the resting level, whereas inositol-1,4,5-trisphosphate generation was still markedly stimulated. Our results demonstrate that an increase in the concentration of cytosolic Ca2+ is not a necessary intermediate between receptor activation and phosphoinositide hydrolysis, and therefore support the second messenger role of inositol-1,4,5-trisphosphate. PMID- 2984217 TI - Identification of a PDGF-like mitoattractant produced by NIH/3T3 cells after transformation with SV40. AB - It has previously been shown that fibroblastic cells transformed by SV40 exhibit a reduced requirement for PDGF for growth. In addition, NIH/3T3 cells lose both their chemotactic response to PDGF and specific cell surface binding of PDGF after transformation with SV40. We have now examined whether the SV40 transformed NIH/3T3 cells are producing a factor which acts similarly to PDGF. Our studies indicate that NIH/3T3 cells transformed with SV 40 produce a factor which shares many biological properties with PDGF. We were unable to detect this activity in conditioned media from nontransformed NIH/3T3 cells. The SV40/NIH/3T3 derived factor appears to possess both chemotactic and mitogenic activity for connective tissue cells but not endothelial or epithelial cells. Furthermore, in preliminary studies, this activity competes with 125I-PDGF for binding to smooth muscle cells. The biochemical properties of the SV40/NIH/3T3 derived factor are different from those of PDGF. The SV40 activity appears to reside in a heat labile acidic protein (pI less than 7.0) of MW less than 30,000 whereas PDGF is a heat stable basic protein (pI9.8) of 30,000 MW. Production of this factor may play a role in the decreased serum requirement for cell replication exhibited by SV40-transformed NIH/3T3 cells by supplying the cells with their own PDGF-like growth factor. PMID- 2984218 TI - Estrogen receptor expression in serially cultivated rat endometrial cells: stimulation by forskolin and cholera toxin. AB - Serially propagated with 3T3 feeder layer support, epithelial cells derived from normal rat endometrium expressed estrogen receptor activity. Specific binding of 17-beta-estradiol was in the range of 30-60 fmol/mg of protein and was of high affinity (Kd = 0.3 nM). A survey of cell lines derived from several other normal epithelia showed that rat vaginal and human cervical cultures also had high affinity estrogen receptors (6-13 fmol/mg of protein), while rat epidermal and esophageal cells had no detectable activity. In the endometrial cultures, receptor levels were elevated nearly two- to fourfold by cholera toxin or forskolin in the medium. This effect was detectable after 4 hr but not 1 hr of treatment and did not occur in the presence of cycloheximide. We conclude that serially cultivated rat endometrial cells retain hormonal properties expressed in vivo while exhibiting some keratinocyte character. These cells may provide a useful model for study of receptor modulation. PMID- 2984219 TI - Serum-free conditions for the growth of avian granulocyte and monocyte clones and primary leukemic cells induced by AMV, and the apparent conversion of granulocytic progenitors into monocytic cells by a factor in chicken serum. AB - The supportive activity of chicken serum for the soft-agar growth of chicken granulocyte and monocyte clones could be replaced with defined ingredients [bovine serum albumin (BSA), conalbumin, selenium, linoleic acid and for routine work, a liquid soy lecithin preparation (59% phospholipids, 39% linoleic acid, and less than 2% of inositol and choline)]. The lecithin preparation could be replaced with L-alpha-phosphatidylcholine. The source of colony-stimulating factor was medium conditioned by fibroblasts cultured under protein-free conditions. AMV-induced leukemic cells could be cloned under identical conditions. In the presence of both chicken serum (10%) and the replacement ingredients, most of the proliferative clones produced were monocytic (84%). In the presence of serum alone, all of them were monocytic. Under serum-free conditions, all the clones produced were granulocytic when a day-3 conditioned medium (CM) was used; monocytic ones were also present when a day-6 CM was used. When the serum was serially diluted in the presence of the replacement ingredients, the number of proliferative monocytic clones progressively decreased while the number of proliferative granulocytic clones progressively increased and the kinetics of each were essentially the same, only opposite in direction. Moreover, the total number of proliferative clones did not change more than 33% at any dilution (or in the absence of serum). We postulate the existence in the chick system of a serum macrophage differentiation factor (M-DF) which converts early granulocytic progenitors (or exclusively diverts a common progenitor) into monocytic cells. PMID- 2984220 TI - Antagonistic effects of insulin and dexamethasone on glucose-regulated and heat shock protein synthesis. AB - The present study extends our previous observation (Kasambalides and Lanks, J. Cell. Physiol., 114:93-98, 1983), that dexamethasone inhibits the alterations in heat shock protein (HSP) and glucose-regulated protein (GRP) synthesis caused by glucose deprivation. We now show that insulin, even in the presence of high extracellular glucose concentrations, will induce 95K and 82K GRP synthesis while suppressing 85K and 69K HSP synthesis. Heat shock of insulin-treated cultures causes induction of the 82K GRP rather than the 85K and 69K HSP's. All of the insulin effects are antagonized by dexamethasone. These data suggest that the changes in GRP and HSP synthesis induced by glucose deprivation and heat shock, respectively, may reflect the operation of a normal physiological mechanism that regulates glucose metabolism. PMID- 2984221 TI - Regulation of gonadotropin release, GnRH receptors, and gonadotrope responsiveness: a role for GnRH receptor microaggregation. PMID- 2984222 TI - Modulation of type alpha transforming growth factor receptors by a phorbol ester tumor promoter. AB - Epidermal growth factor (EGF) and an EGF-like transforming growth factor (eTGF) from retrovirally transformed cells bind to a common receptor type in A431 cells. We have investigated the effects of the tumor promoter phorbol myristate acetate [PMA] on EGF/eTGF receptors in intact A431 cells. Treatment with PMA at 37 degrees C induces a complete loss of high-affinity (Kd = 35-50 pM) binding sites for eTGF and EGF on the cell surface of A431 cells. This effect is half-maximal at 0.1 nM PMA, exhibits rapid kinetics, and persists for at least 4 hr in the presence of PMA. eTGF and PMA added to intact A431 cells induce the phosphorylation of immunoprecipitable 170kd EGF/eTGF receptors. The EGF/eTGF receptor isolated from control cells was found to contain phosphoserine and phosphothreonine. PMA and eTGF caused a marked increase in the level of these two phosphoamino acids. In addition, eTGF but not PMA caused the appearance of phosphotyrosine in the EGF/eTGF receptor in vivo. We conclude that the tumor promoting phorbol diester regulates both the affinity and phosphorylation state of the A431 cell receptor for the type alpha transforming growth factors, eTGF and EGF. PMID- 2984223 TI - [Secretion of gastric acid, gastrin and cyclic nucleotides in relation to the action of the vagus nerve]. AB - Changes on levels gastric acidity, on serum gastrin, cAMP and cGMP levels were studied in 18 healthy volunteers after either stimulation (injection of 0.2 IU of soluble insulin/kg body weight to a group of 9 subjects) or stimulation and inhibition (injection of insulin plus atropine per os to the other group of 9 subjects) of the vagus nerve. After vagus nerve stimulation, gastric acid levels, serum gastrin and cGMP were raised and cAMP reduced. After stimulation and inhibition of vagus nerve, gastrin and cAMP were increased, cGMP reduced and gastric acid levels remained unchanged. These findings suggest that the vagus nerve, and more particularly the acetylcholine released from its metaganglionic fibers, stimulate parietal cells provoking acid secretion, and also stimulate G cells with subsequent gastrin secretion through cGMP. PMID- 2984224 TI - [Conservative treatment of cancer of the breast. Apropos of 108 cases]. AB - Between 1975 and 1980, 108 cancers of the breast (T1 and small T2) were treated by associated conservative surgery and radiotherapy. Axillary glands were curetted in 92 patients; 17% showed lymph node invasion. Radiotherapy was by telecobalt: 45 grays to the whole breast and 60 grays to the tumor bed. Esthetic results were satisfactory. Intramammary recurrence was rare (6%) and was independent of site or size of tumor and of degree of lymph node involvement. Metastases not associated with local recurrence were very rarely observed (4% of cases) and were present only in patients without glandular involvement. The 5 year actuarial survival rate was 95%. PMID- 2984225 TI - [A frequent malignant tumor of the soft tissues, malignant fibrohistiocytoma. Current interest: apropos of 2 cases disclosed by vascular complications]. AB - Malignant fibrohistiocytoma represents one of the most common malignant tumors of soft tissues. Previously difficult to diagnose with precision, it can now be detected pathologically as a result of recent techniques for investigating cellular development. Treatment is by wide exeresis, prognosis remaining reserved and requiring prolonged postoperative surveillance. PMID- 2984226 TI - Reversed-phase chromatography of proteins with diphenyl-silica column and hydro organic eluents containing two organic solvents. AB - High-performance liquid chromatography of proteins with silica bonded diphenyl stationary phase and hydro-organic mobile phases with linear gradient elution has been carried out with binary organic modifiers. The use of a mixture of 2 propanol and 1-butanol facilitated elution at total organic modifier concentrations significantly lower than with the use of 2-propanol alone. Furthermore higher protein recovery and retention of biological activity was obtained with the binary organic modifier because the increase in eluent strength with binary organic modifier was greater than that of the denaturing strength of the eluent. The use of a short, 3 cm long, column, relatively high flow-rates and steep gradients was also advantageous in attenuating protein denaturation. The results were interpreted within the framework of a theoretical treatment of the combined effect of the retention process and denaturation reaction simultaneously occurring in the column. PMID- 2984227 TI - Estimation of DNA fragment size and generation of DNA restriction endonuclease maps using linear models. AB - A method for the estimation of DNA fragment size and for the generation of DNA restriction endonuclease maps using linear models is discussed, and a computer program which utilizes the SAS (SAS Institute Inc., 1982) statistical package is presented. PMID- 2984228 TI - Virus enzyme-linked cell immunoassay (VELCIA): detection and titration of rotavirus antigen and demonstration of rotavirus neutralizing and total antibodies. AB - Virus enzyme-linked cell immunoassay (VELCIA) for detection and titration of rotavirus antigen has been developed. Wild-type porcine rotavirus antigen can be detected and titrated directly from the fecal material within 24 h. Porcine OSU strain can be titrated higher than 10(-8). The method has also been introduced for the demonstration of rotavirus neutralizing and total antibodies. In VELCIA the advantages of the cell culture system for virus isolation are combined with enzyme immunodetection and spectrophotometrical reading of the test. PMID- 2984230 TI - A method for detecting human enteroviruses in aquatic sediments. AB - A method is described for detecting enteroviruses in both freshwater and marine sediments. Viruses were recovered from sediments by elution into 6% beef extract at pH 9.0 and concentration with polyethylene glycol 6000. The recovery efficiency ranged from 6 to 55% for marine sediments and 16 to 77% for freshwater sediments. Although the efficiency of the method was influenced by the composition and source of the sediments it was used successfully to detect viruses occurring in marine and freshwater sediments near sewer outfalls. PMID- 2984229 TI - Rocket line immunoelectrophoresis: an improved assay for simultaneous quantification of a mink parvovirus (Aleutian disease virus) antigen and antibody. AB - A rocket line immunoelectrophoretic assay (RLIE) was developed for the simultaneous quantification of viral antigens and antiviral antibodies of the important mink parvovirus, Aleutian disease virus (ADV). The sensitivity of the RLIE assay was found to be 5 log2 higher than that of the counter current immunoelectrophoresis which is the assay routinely used for diagnostic purposes. PMID- 2984231 TI - The immune response to poliovirus-specific synthetic peptides: effects of adjuvants and test animal species. AB - Carrier protein conjugates of five synthetic peptides containing amino acid sequences specific to capsid proteins VP1 and VP2 of poliovirus type 1 were tested for their abilities to elicit an immune response in the presence of either of two adjuvants and in several animal species. Freund's adjuvant induced significantly higher level anti-peptide antibody titers than A1(OH)3. However, no difference was noted between the two adjuvants in their abilities to aid in the induction of cross-reactive virus neutralizing antibody. The latter antibody was more readily produced by rabbits than by guinea pigs in spite of equivalent anti peptide titers. Rats failed to produce neutralizing antibodies and their anti peptide antibody levels were generally lower. The significance of these results for studies involving the development of synthetic peptide immunogens is discussed. PMID- 2984232 TI - Use of the dexamethasone-adrenocorticotropin test to assess the requirement for continued glucocorticoid replacement therapy after pituitary surgery. AB - We assessed the need for continued glucocorticoid replacement therapy in postsurgical pituitary tumor patients using a dexamethasone-ACTH test. The patients received 1 mg dexamethasone, orally, at 2300 h and 250 micrograms synthetic ACTH (Cosyntropin), iv, at 0800 h the next morning. The mean +/- SD integrated cortisol response for a 2-h period of the 31 pituitary tumor patients [1264 +/- 924 micrograms X min/dl (34.87 +/- 25.49 mumol X min/liter)] was significantly less (P less than 0.005) than that of 25 normal subjects [3331 +/- 544 micrograms X min/dl (91.90 +/- 17.04 mumol X min/liter)]. Replacement glucocorticoids were abruptly discontinued in 11 patients with responses above 1450 micrograms X min/dl (40.01 mumol X min/liter). No clinical or laboratory evidence of adrenal insufficiency occurred as long as 15 months after discontinuation. Metyrapone tests, however, in the 11 glucocorticoid-withdrawn patients revealed a reduced mean +/- SD serum 11-deoxycortisol level compared with that of 10 normal subjects [8.9 +/- 4.7 vs. 15.6 +/- 5.0 micrograms/dl (0.26 +/- 0.13 vs. 0.45 +/- 0.16 mumol/liter); P less than 0.005]. Our results indicate that the dexamethasone-ACTH test is useful in identifying patients in whom replacement glucocorticoid therapy can be safely withdrawn under nonstressed conditions. The test can be simplified to one plasma cortisol level determined 30 min after ACTH administration. PMID- 2984233 TI - Effect of subcutaneous and intranasal administration of ovine corticotropin releasing hormone in man: comparison with intravenous administration. AB - Long term use of ovine corticotropin-releasing hormone (oCRH) requires a convenient route of administration. The effects of 0.3, 3, and 30 micrograms/kg BW synthetic oCRH given as a sc injection and of 10 and 30 micrograms/kg given as an intranasal spray were studied in 10 normal men in the late afternoon. Basal plasma immunoreactive ACTH (IR-ACTH) and IR-cortisol levels were 14 +/- 1.9 pg/ml and 4.3 +/- 0.4 microgram/dl (mean +/- SEM). Peak IR-ACTH levels (mean +/- SEM) were 43 +/- 5.5, 53 +/- 8.1, and 64 +/- 8.9 pg/ml after the 0.3, 3, and 30 micrograms/kg doses of oCRH given sc, respectively, and 23 +/- 4.3 and 36 +/- 4.8 pg/ml after the 10 and 30 micrograms/kg doses of oCRH given intranasally, respectively. The lowest sc dose and both intranasal doses caused only single IR ACTH peaks. After 3 and 30 micrograms/kg sc oCRH, IR-ACTH rose by 15 min, reached an initial peak at 45-60 min, fell rapidly until 90-120 min, and rose to a second peak at 3-5 h. This biphasic response is similar to that previously found after iv administration. IR-ACTH levels remained elevated for 4, 10, and at least 16 h after 0.3, 3, and 30 micrograms/kg sc oCRH, respectively, and for 1.5 and 3 h after 10 and 30 micrograms/kg intranasal oCRH respectively. The effect on IR cortisol was similar, but more prolonged. Compared to the iv route, sc oCRH produced similar mean peak IR-ACTH and IR-cortisol levels and had a slightly longer duration of action. Intranasal oCRH was only about 1% as effective. Peak plasma IR-oCRH levels in 2 subjects receiving 3 micrograms/kg sc oCRH were 13 and 17 ng/ml at 90 min. These peaks were lower than those after iv administration of the same dose, but the levels remained elevated longer, probably accounting for the longer duration of action of sc oCRH. Peak plasma IR-oCRH levels in 4 subjects given 10 microgram/kg intranasal oCRH were only 64-122 pg/ml, presumably reflecting poor absorption through the nasal mucosa. These results demonstrate that sc injection of oCRH is at least as effective as the iv route with respect to plasma IR-ACTH and IR-cortisol responses. The convenience of this route of administration and the prolonged duration of action of oCRH suggest the feasibility of long term oCRH use. PMID- 2984234 TI - In vitro metabolism of 25-hydroxycholecalciferol by isolated cells from human decidua. AB - An increase in maternal serum levels of 1 alpha, 25-dihydroxycalciferol during pregnancy has been linked to enhancement of intestinal calcium absorption. Several sites of its synthesis have been proposed in different species, human decidua being one of them. Collagenase-dispersed decidual cells isolated from term placenta were fractionated on a Percoll gradient. The isolated cells were set in culture in the presence of 6 nM [3H]cholecalciferol. Two cell populations of similar morphology hydroxylated the substrate, yielding a compound that had a mass spectrum identical to and that comigrated with authentic 1 alpha, 25 dihydroxycholecalciferol in four chromatographic systems and bound to a specific rachitic chick receptor. These preparations, thus, provide a potential system by which the kinetics and regulation of the synthesis of the hormonal form of vitamin D by human placenta can be studied in vitro. PMID- 2984235 TI - Morphine inhibits the pituitary-adrenal response to ovine corticotropin-releasing hormone in normal subjects. AB - To determine the locus of opiate modulation of ACTH secretion, 11 normal subjects were given ovine corticotropin-releasing hormone (CRH) 30 min after receiving either placebo or morphine sulfate. Plasma ACTH, cortisol, arginine vasopressin (AVP), epinephrine, norepinephrine, and CRH were measured 30 min before and up to 150 min after CRH administration. Morphine blunted the ACTH response for the first 60 min and cortisol response for the first 90 min after CRH administration. Morphine did not lower arginine vasopressin or catecholamine levels. To determine whether morphine's effect on ACTH and cortisol was due to a direct action on the corticotroph cell, dispersed rat pituitary cells were perifused with medium containing 1 microgram/ml morphine sulfate or medium alone. Morphine had no effect on the ACTH response of these cells to 10 nM CRH pulses. Similarly, morphine had no effect on ACTH production by dispersed rat pituitary cells in monolayer culture in response to 90- and 180-min incubations with 5 nM CRH. We conclude that morphine blunts the early response of the pituitary gland to CRH in vivo. Based on the lack of a direct effect of morphine on rat pituitary cells in vitro, we postulate that morphine given in vivo may modulate the pituitary ACTH response to CRH through other suprapituitary factors. PMID- 2984236 TI - Ectopic beta-adrenergic receptors coupled to adenylate cyclase in human adrenocortical carcinomas. AB - The adenylate cyclase of an adrenocortical carcinoma of the rat is activated not only by ACTH but also by beta-adrenergic agonists, which bind to ectopic beta adrenergic receptors not present in normal rat adrenal cortex. Previous reports examining possible beta-adrenergic control of adenylate cyclase in human adrenocortical carcinomas failed to demonstrate beta-adrenergic receptor-linked enzyme activity. We studied six human adrenal carcinomas and normal adrenal cortex from three subjects for beta-adrenergic agonist-sensitive adenylate cyclase and beta-adrenergic binding sites. Three of the six carcinomas had adenylate cyclase responses to both ACTH and beta-agonists. Two tumors were ACTH responsive but not beta-agonist responsive; one tumor responded to beta-agonists but not to ACTH. Adenylate cyclase activity of normal adrenal cortex from three subjects was stimulated by ACTH but not by beta-agonists. In membrane preparations from three tumors with beta-agonist-sensitive adenylate cyclase, the radiolabeled beta-adrenergic antagonist [125I]pindolol bound specifically and with high affinity (Kd = 38-83 pM) to a single class of binding sites which showed saturation with ligand concentration, reversibility of binding, pharmacological specificity, and stereospecificity. Normal cortex and one tumor without beta-adrenergic agonist-sensitive adenylate cyclase had no specific binding of [125I]pindolol. These results indicate that malignant transformation of adrenal cortex in man is frequently but not invariably associated with the appearance of ectopic beta-adrenergic receptors functionally linked to adenylate cyclase. Loss of ACTH-responsive adenylate cyclase may also occur simultaneously with the development of beta-adrenergic receptor-linked adenylate cyclase. PMID- 2984237 TI - The vasopressin-sensitive adenylate cyclase in collecting tubules and in thick ascending limb of Henle's loop of human and canine kidney. AB - The major tubular effects of [8-Arg]vasopressin (AVP) in regulation of renal water excretion are initiated by stimulation of adenylate cyclase (AdC) coupled with V2 receptors. We explored whether the AVP-sensitive AdC is present in both collecting tubules and the thick ascending limb of Henle's loop of human and canine kidney. In cortical collecting tubule (CCT) and medullary collecting tubules (MCT) of human kidney, AdC was markedly stimulated by AVP [maximum change from basal level (delta), +2700%] and the the nonhormonal stimulatory agent forskolin (delta, +2000%). In human CCT, the effects of both compounds were synergistic. In contrast, AVP had no effect on AdC in either the medullary (MAL) or cortical (CAL) segment of the thick ascending limb of Henle's loop of human kidney; AVP also did not stimulate AdC in CAL or MAL in the presence of forskolin. Similar to that in the human kidney, in the canine kidney, AdC in CCT and MCT was markedly stimulated by AVP and forskolin (delta, +1000%), but AVP had no effect on AdC in CAL and MAL of the canine kidney. In intact tubules dissected from dog kidney and incubated in vitro, AVP markedly increased cAMP accumulation in MCT. AVP also elicited a small but detectable increase in cAMP accumulation in MAL. From these observations, we conclude that AVP-sensitive AdC is well developed in collecting tubules, but that AVP-sensitive AdC is absent in MAL and CAL of human kidney. Likewise, in canine nephron, the AVP-sensitive AdC of MAL and CAL is rudimentary or very labile. These findings suggest that the unresponsiveness of the AdC-cAMP system to AVP in segments of the thick ascending limb of Henle's loop may be a factor that accounts for a relatively low maximum osmotic concentration of urine which can be achieved by human or canine kidneys. PMID- 2984239 TI - An effect of gene dosage on production of human chorionic somatomammotropin. AB - The gene deletions responsible for isolated partial deficiency of fetal human chorionic somatomammotropin (hCS) production were characterized by restriction endonuclease analysis of genomic DNA prepared from the leukocytes of an affected child. The phenotypically normal child was the product of a 38-week pregnancy characterized by peak maternal hCS levels of 1.1 micrograms/microliter (normal, 3 9.2 micrograms/ml) and normal levels of other pregnancy-associated hormones. Two genes, termed hCS-A and hCS-B, specify the same mature hCS peptide and are responsible for fetal hCS production. Digestion of the child's DNA with the enzymes Hind III, Eco RI, Bam HI, Bgl II, Hinc II and Msp I disclosed absence of the restriction fragments that normally contain the hCS-A gene. However, the hCS B gene was present in the child's DNA. The child's DNA digests contained an abnormally large Eco RI fragment of 10.0 kb containing the hCS-L gene. This abnormal fragment is a marker for a deletion that is responsible for complete deficiency of hCS when present in the homozygous state. The child's DNA restriction patterns were consistent with heterozygosity for two different deletions involving hCS genes. The paternal hGH gene cluster lacked the hCS-A, human GH variant, and hCS-B genes, while the maternal cluster lacked only the hCS A gene. Thus, the child's DNA contained only one of the normal complement of four functional hCS genes per diploid genome. Material hCS levels approximately one fourth as great as those present at comparable stages of normal pregnancies indicated that there was no compensatory increase in expression of the remaining hCS gene. PMID- 2984238 TI - Isolation and structural identification of 1,25-dihydroxyvitamin D3 produced by cultured alveolar macrophages in sarcoidosis. AB - Hypercalcemia and hypercalciuria in sarcoidosis are thought to result from the endogenous overproduction of an active vitamin D metabolite. We employed primary cultures of pulmonary alveolar macrophages from two patients with biopsy-proven pulmonary sarcoidosis and a recent or current clinical abnormality in calcium metabolism to synthesize in vitro a 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]-like metabolite from 25-hydroxyvitamin D3 (25OHD3). The macrophage metabolite cochromatographed with [3H]1,25-(OH)2D3 on normal phase and reverse phase high performance liquid chromatography and was bound with high affinity by the chick intestinal receptor for 1,25-(OH)2D3. On UV spectroscopy, the metabolite possessed the carbon-5,7,10 (19) cis-triene chromophore characteristic of a vitamin D sterol. Electron impact mass spectrometry of trimethylsilyl ether derivatives of the metabolite revealed a mass fragmentation pattern similar to that of the trimethylsilyl ether derivative of authentic 1,25-(OH)2D3. The incubation of cultured macrophages from two patients with idiopathic pulmonary fibrosis and two with scleroderma with [3H]25OHD3 did not result in production of a metabolite with the chromatographic identity of 1,25-(OH)2D3. These data indicate that the metabolite of 25OHD3 synthesized by sarcoid macrophages in vitro is 1,25-(OH)2D3 and that the macrophage is a synthetic source of the sterol metabolite in sarcoidosis. PMID- 2984240 TI - Recognition of Staphylococcus saprophyticus in urine cultures by screening colonies for production of phosphatase. AB - Phenolphthalein diphosphate was incorporated into a primary blood agar medium for use in performing quantitative urine cultures. Phosphatase-negative staphylococci, such as Staphylococcus saprophyticus, were differentiated from phosphatase-positive species, such as Staphylococcus epidermidis, by spot testing colonies on filter paper saturated with 1 N NaOH. Phosphatase-positive colonies turned pink within seconds, and phosphatase-negative colonies showed no color. None of 55 S. saprophyticus isolates showed production of phosphatase on this medium. Of 193 consecutive coagulase-negative staphylococci isolated from the urine of 190 adolescent female patients, 84% were phosphatase positive, non-S. saprophyticus species; 16% were phosphatase-negative and indicated S. saprophyticus (22), Staphylococcus haemolyticus (4), Staphylococcus simulans (2), Staphylococcus warneri (1), and Staphylococcus hominis (1). Phosphatase activity was variable in the other flora encountered in the urine cultures. Mixtures of phosphatase-positive and -negative organisms did not cause false-positive reactions. PMID- 2984241 TI - Comparison of a latex agglutination test with five other methods for determining the presence of antibody against cytomegalovirus. AB - A latex agglutination test for determination of antibody against cytomegalovirus was compared with five other methods: a solid-phase fluorescent immunoassay, an indirect hemagglutination test, two solid-phase enzyme immunoassays, and an indirect fluorescent-antibody method, with sera collected from 210 random blood donors. Of the sera tested, 28% were positive for anti-cytomegalovirus by concordance of four or more methods. The latex agglutination test performed well, with a sensitivity of 100%, a specificity of 99%, and positive and negative predictive values of 97 and 100%, respectively. The methods were also evaluated for the number of sera requiring repeat testing, equivocal results after retesting, ease of performance, turnaround time, and technical demands. The tests which best met the requirements for a screening test were the solid-phase fluorescent immunoassay, the indirect hemagglutination test, and the latex agglutination test. The latex agglutination test is a valuable screening tool for detecting total anti-cytomegalovirus which has high sensitivity, high negative predictive value, and rare equivocal results and also has the added advantages of ease of performance and rapid turnaround time. PMID- 2984242 TI - Microcarriers in combination with enzyme immunofiltration and immunofluroescence for the detection of herpes simplex virus antigens in culture. AB - A microcarrier culture system in combination with enzyme immunofiltration with a herpes simplex virus (HSV) group monoclonal antibody was found to be as sensitive as immunofluorescence for the detection of HSV type 1 (HSV-1) in cell cultures and to give specific identification at the same time as the appearance of the cytopathic effect with very high infectious inocula and within 10 to 24 h of the appearance of the cytopathic effect with very high infectious inocula and within 10 to 24 h of the appearance of the cytopathic effect with low to high inocula. Multiplicities of infection from 10(-5) to 10(1) were tested at 8 to 96 h postinfection. When applied to the identification of HSV-1 and HSV-2 in cultures of clinical samples, the system detected HSV antigens in 50% of the samples after 2 days and in 100% of the samples after 3 days. With 2 ml of microcarrier suspension and with 50 to 300 microliter per sample, several portions are available for replicate and sequential sampling without destroying the culture. The system requires only that 2 X 10(5) microcarriers be added to the culture tube at the time or before it is seeded with cells, at an extra cost of 6 cents (U.S.) per tube and little extra labor. PMID- 2984243 TI - Porcine rotavirus-like virus (group B rotavirus): characterization and pathogenicity for gnotobiotic pigs. AB - A rotavirus-like virus (RVLV) was isolated from a diarrheic pig from an Ohio swine herd. This virus infected villous enterocytes throughout the small intestine of gnotobiotic pigs and induced an acute, transitory diarrhea. Complete virions were rarely observed in the intestinal contents of infected animals; the predominant particle detected by immune electron microscopy was a corelike particle 52 nm in diameter. The genome of the porcine RVLV was composed of 11 discrete segments of double-stranded RNA that produced an electropherotype distinct from the genome electropherotypes of reovirus, rotavirus, and porcine pararotavirus. Porcine RVLV was antigenically unrelated to rotavirus, porcine pararotavirus, or reovirus but was antigenically related to a bovine RVLV. PMID- 2984245 TI - Comparison of genomic homologies in the coxsackievirus B group by use of cDNA:RNA dot-blot hybridization. AB - Radioactive cDNAs to coxsackievirus B (CB) genomic RNAs were used to probe genomic homologies between RNAs of the CB group serotypes. CB5 shared approximately equivalent homology with the other five CB serotypes. CB1, CB2, and CB3 hybridized preferentially to CB5 in turn, suggesting that these three have diverged in one direction from CB5. CB4 and CB6 were more related to each other than to the other serotypes at a lower criterion, but at a more stringent criterion they did not demonstrate apparent homology with each other. This suggests that they have diverged from CB5 as well but perhaps earlier in time or at a greater rate than did CB1, CB2, or CB3. Some potential applications of these relationships to clinical CB detection assays based on nucleic acid hybridization are discussed. PMID- 2984244 TI - Detection of Epstein-Barr virus DNA in mouthwashes by hybridization. AB - An assay for the presence of Epstein-Barr virus (EBV) DNA was developed by using a cloned EBV DNA probe. After preliminary testing showed the assay to be sensitive and specific, it was applied to 135 mouthwashes from bone marrow transplant recipients, and 21 of these tests were positive. The concentration of EBV DNA in mouthwashes in some cases was as high as 10(8) genome equivalents per ml. When compared with the lymphocyte transformation assay on the same samples, the sensitivity was 75% and the specificity was 97%. In contrast to the lymphocyte transformation assay, the hybridization was semiquantitative and yielded results in 72 h. Potential applications include monitoring the effects of various interventions, such as immunosuppressive and antiviral chemotherapy, on EBV shedding. PMID- 2984246 TI - Rapid detection of bovine herpesvirus type 1 antigens in nasal swab specimens with an antigen capture enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of bovine herpesvirus type 1 (BHV-1) antigens in nasal swab specimens collected from infected animals. Development of the ELISA involved screening and selection of BHV-1-specific monoclonal antibodies for their ability to capture BHV-1 antigens and for their stability and activity after conjugation to horseradish peroxidase. Forty combinations of capture-conjugate monoclonal antibody pairs were screened for detection of nanogram amounts of purified BHV-1 by using a double-antibody sandwich ELISA in which antigen and conjugated antibody were simultaneously added to antibody-coated wells. Of the 40 monoclonal antibody pairs, 4 were analyzed further and 1 was selected for routine application to clinical specimens. Of 129 nasal swab specimens collected during the first 10 days after experimental infection with BHV-1, 66 were found to be positive by both virus isolation and ELISA and 34 were positive for infectious virus but negative by ELISA. One specimen was positive by ELISA but negative by virus isolation, and the remaining 28 specimens were negative by both tests. Quantitation of the virus-containing specimens showed that the ELISA had a lower detection limit of 10(3.5) median tissue culture infective doses. The ELISA was judged to be highly useful for diagnosis of BHV-1 infections, since all of the nasal swab specimens that were collected from 12 animals during the first 5 days of the infection, when the clinical signs were the most apparent, were positive. PMID- 2984248 TI - Evaluation of a commercial enzyme-linked immunosorbent assay for the detection of herpes simplex virus. AB - A total of 136 specimens were tested for the presence of herpes simplex virus by routine tissue culture and a commercial enzyme-linked immunosorbent assay (Ortho Diagnostic Systems, Inc.). Forty-six (33.8%) of the specimens were positive by tissue culture. The sensitivity and specificity of the commercial system were 69.6 and 93.3%, respectively. The commercial system was rapid and moderately specific but lacked the sensitivity necessary for direct specimen testing. PMID- 2984247 TI - Serotypic characterization of rotaviruses derived from asymptomatic human neonatal infections. AB - Nineteen rotavirus strains derived from asymptomatic neonates (seven from England, five from Australia, two from Venezuela, and five from Sweden) were successfully cultivated in primary African green monkey kidney cell cultures, serotyped by plaque reduction neutralization tests, subgrouped by indirect enzyme linked immunosorbent assay, and electropherotyped by polyacrylamide gel electrophoresis. All 19 strains were shown to fall into one of the four known human serotypes; serotype 1 (all Venezuelan strains), serotype 2 (all Swedish strains), serotype 3 (all Australian strains), or serotype 4 (all English strains). Hyperimmune guinea pig serum raised against the Venezuelan strain (M37) neutralized not only serotype 1 (strain Wa) but also serotype 4 (strain St. Thomas no. 3) viruses to a similar degree. The English, Australian, and Venezuelan isolates were found to belong to subgroup 2, and the Swedish strains were subgroup 1 viruses. The potential importance of these rotaviruses obtained from neonates as possible vaccine candidates is discussed. PMID- 2984249 TI - Problems associated with the use of (E)-5-(2-bromovinyl)-2'-deoxyuridine for typing herpes simplex virus. AB - When 0.5 microgram of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVdU) per ml was incorporated directly into cell culture medium inoculated with eight known positive specimens, one herpes simplex virus type 1 (HSV-1) isolate grew in the presence of BVdU and was misidentified. By plaque assay, the titers of 15 HSV-1 strains were reduced by more than 3 log10 by BVdU, and the titers of 16 HSV-2 strains were reduced by less than 2 log10. Titers of HSV-1 acyclovir-resistant strains were reduced by less than 1.5 log10, which was characteristic of HSV-2 strains. Thus, typing of HSV isolates in the presence of BVdU by plaque assay is reliable only if information regarding previous antiviral therapy is obtained. PMID- 2984250 TI - Gelsolin, a Ca2+-dependent actin-binding protein in a hamster insulin-secreting cell line. AB - Using a gel overlay technique we have previously described a 90,000-mol wt actin binding protein in a number of hormone-secreting tissues and tentatively identified this protein as gelsolin. Gelsolin is a protein that cuts or solates cross-linked actin filaments and can also serve as a nucleating site for actin polymerization. The objective of this study was to isolate this protein from a hamster insulin-secreting (HIT) cell line and compare the immunologic properties and peptide maps of purified rabbit macrophage gelsolin, human platelet gelsolin, and the HIT cell 90,000-mol wt protein. DNase I-Sepharose retained the HIT cell actin-binding proteins in 1 mM CaCl2; some of the 90,000-mol wt protein could then be eluted with 1 mM EGTA. The remaining actin-binding proteins were eluted using a buffer containing SDS. The EGTA peak fractions contained two major protein bands of Mr = 90,000 and 42,000, which suggested that a 90,000-mol wt actin complex was eluted from the DNase I-Sepharose column. Specific antibodies to the human platelet and rabbit macrophage gelsolins bound to the 90,000-mol wt bands in the eluates, but did not crossreact with other actin-binding proteins. Indirect immunofluorescence using an anti-human platelet gelsolin antibody localized the 90,000-mol wt protein to stress fibers that were also stained with phalloidin, which suggested that gelsolin is associated with actin in vivo. Tryptic peptide maps of all three radioiodinated gelsolins were virtually indistinguishable. Thus, gelsolin is a highly conserved gene product found in at least three diverse cell types, an insulin-secreting beta cell line, macrophages, and platelets, and may link a transient increase in Ca2+ cellular levels with changes in actin polymerization and/or the gel-sol state of these cells. PMID- 2984251 TI - Receptors and growth-promoting effects of insulin and insulinlike growth factors on cells from bovine retinal capillaries and aorta. AB - It has been suggested that elevated levels of insulin or insulin-like growth factors (IGFs) play a role in the development of diabetic vascular complications. Previously, we have shown a differential response to insulin between vascular cells from retinal capillaries and large arteries with the former being much more insulin responsive. In the present study, we have characterized the receptors and the growth-promoting effect of insulinlike growth factor I (IGF-I) and multiplication-stimulating activity (MSA, an IGF-II) on endothelial cells and pericytes from calf retinal capillaries and on endothelial and smooth muscle cells from calf aorta. We found single and separate populations of high affinity receptors for IGF-I and MSA with respective affinity constants of 1 X 10(-9) M-1 and 10(-8) M-1 in all four cell types studied. Specific binding of IGF-I was between 7.2 and 7.9% per milligram of protein in endothelial cells and 9.1 and 10.4% in the vascular supporting cells. For 125I-MSA, retinal endothelial cells bound only 1.7-2.5%, whereas the aortic endothelial cells and the vascular supporting cells bound between 5.6 and 8.5% per milligram of protein. The specificity of the receptors for IGF-I and MSA differed, as insulin and MSA was able to compete with 125I-IGF-I for binding to the IGF-I receptors with 0.01-0.1, the potency of unlabeled IGF-I, whereas even 1 X 10(-6) M, insulin did not significantly compete with 125I-MSA for binding to the receptors for MSA. For growth-promoting effects, as measured by the incorporation of [3H]thymidine into DNA, confluent retinal endothelial cells responded to IGF-I and MSA by up to threefold increase in the rate of DNA synthesis, whereas confluent aortic endothelial cells did not respond at all. A similar differential of response to insulin between micro- and macrovascular endothelial cells was reported by us previously. In the retinal endothelium, insulin was more potent than IGF-I and IGF-I was more potent that MSA. In the retinal and aortic supporting cells, no differential response to insulin or the IGFs was observed. In the retinal pericytes, IGF-I, which stimulated significant DNA synthesis beginning at 1 X 10( 9) M, and had a maximal effect at 5 X 10(-8) M, was 10-fold more potent than MSA and equally potent to insulin. In the aortic smooth muscle cells, IGF-I was 10 100 times more potent than insulin or MSA. In the retinal and aortic supporting cells, no differential response to insulin or the IGFs was observed. In the retinal pericytes, IGF-I, which stimulated significant DNA synthesis beginning at 1 X 10(-9) M, and had a maximal effect at 5 X 10(-8) M, was 10-fold more potent than MSA and equally potent to insulin. In the aortic smooth muscle cells, IGF-I was 10-100 times more potent than insulin or MSA. In addition, insulin and IGF-I at 1 X 10(-6) and 1 X 10(-8) M, respectively, stimulated these cells to grow by doubling the number of cells as well. In all responsive tissues, the combination of insulin and IGFs were added together, no further increase in effect was seen. These data showed that vascular cells have insulin and IGF receptors, but have a differential response to these hormones. These differences in biological response between cells from retinal capillaries and large arteries could provide clues to understanding the pathogenesis of diabetic micro- and macroangiopathy. PMID- 2984252 TI - Dietary and hormonal regulation of aldolase B gene expression. AB - Aldolase B is an enzyme of the glycolytic pathway whose activity and mRNA levels in the liver fluctuate according to dietary status. Both the enzyme activity and the mRNA concentration decline during fasting and increase four- to eightfold upon refeeding of a carbohydrate-rich diet. The mechanism, however, of the mRNA induction remains unknown. To elucidate the mechanisms that regulate this induction responsive to dietary stimuli, we have studied the roles of hormones and glycolytic substrates on aldolase B gene expression in three tissues that synthesize the enzyme. Using a cDNA probe complementary to rat aldolase B mRNA, we determined the amount of cytoplasmic RNAs in the liver, kidney, and small intestine of normal, adrenalectomized, thyroidectomized, diabetic, and glucagon- or cAMP-treated animals refed either a fructose-rich or a maltose-rich diet. The in vivo hormonal control of gene expression was found to be very different in the three organs tested. In the liver, cortisone and thyroid hormones were required for the induction of the specific mRNA by carbohydrates, while in the kidney none of the hormonal modifications tested altered the level of mRNA induction. In the liver, but not in the kidney, diabetes and glucagon administration abolished the induction of aldolase B mRNAs in animals refed the maltose-rich diets. In the small intestine, only diabetes and thyroidectomy affected the gene expression. Finally, no induction occurred when normal fasted rats were given any of the hormones. Thus, the in vivo hormonal control of liver aldolase B gene expression differs significantly from that of kidney and small intestine. In the liver, the mRNA induction requires the presence of dietary carbohydrates, of permissive hormones, and the cessation of glucagon release, while in the kidney, the induction of the mRNAs by fructose occurs regardless of the hormonal status of the animals. The hormonal control of aldolase B mRNA levels in the small intestine is intermediate. PMID- 2984253 TI - Role of transferrin, Fe, and transferrin receptors in myeloid leukemia cell growth. Studies with an antitransferrin receptor monoclonal antibody. AB - In previous studies, antitransferrin receptor antibody 42/6 inhibited growth of normal granulocyte/macrophage progenitors and some malignant myeloid cells. In these studies, leukemia cell lines cultured without serum and fresh leukemia cells were used to investigate the roles of Fe, transferrin receptors, and transferrin in leukemia cell growth, and mechanisms of 42/6 inhibition and resistance. HL60 and KG-1 leukemia cells grown in serum-free medium were inhibited by 42/6. In contrast to results in fetal calf serum (FCS), soluble Fe (ferric nitriloacetate) reversed 42/6 growth inhibition of serum-free HL60 cells. When HL60 cells were adapted for growth in serum-free, transferrin-free medium, they became refractory to 42/6 growth inhibition. By using radiolabeled transferrin and 42/6, HL60 cells cultured in FCS and transferrin displayed similar quantities of transferrin receptors (29,000-30,000/cell) and similar Kd's (3.8-4.9 X 10(-9) M). Cells grown in transferrin-free medium showed a similar Kd (3.1 X 10(-9) M), but fewer transferrin binding sites (5,000/cell). Transferrin independent cells contained a log higher concentration of intracellular ferritin. For both FCS and serum-free HL60 cells, calculated affinities for 42/6 were lower (5.7-10.0 X 10(-9) M), but the number of binding sites was three- to fourfold higher. To investigate further the relationship between receptor display and antibody inhibition in proliferating normal and malignant myeloid cells, simultaneous immunofluorescence was used to determine the cell cycle status of transferrin receptor-positive cells. Malignant cells in S + G2/M displayed approximately 50% of the amount of transferrin receptors detected in normal dividing colony-stimulating factor-stimulated marrow cells. Receptor display by dividing cells from two patients with acute nonlymphocytic leukemia was variable. When HL60 cells were exposed to dimethyl sulfoxide, transferrin receptor display decreased, and 42/6 growth inhibition was abrogated or greatly diminished. The presence of 42/6 did not prevent dimethyl sulfoxide-induced HL60 differentiation in serum-containing or serum-free cultures. We conclude that human leukemia cells require Fe for growth and that 42/6 inhibits transferrin-dependent cells by Fe deprivation. Some dividing normal and differentiating malignant cells display reduced transferrin receptors, and can also escape antibody inhibition. The increased ferritin levels and decreased transferrin receptors in transferrin independent HL60 cells confirm the inverse relationship between cell ferritin content and transferrin receptor display. These studies indicate a critical role for Fe in leukemia cell growth and possible roles in cellular differentiation. PMID- 2984254 TI - Elevated epidermal growth factor receptor gene copy number and expression in a squamous carcinoma cell line. AB - The human epidermal growth factor (EGF) receptor is known to be homologous to the v-erb B oncogene protein of the avian erythroblastosis virus. Overexpression of the EGF receptor gene in A431 epidermoid carcinoma cells is due to gene amplification. In this study, a variety of squamous cell carcinomas were examined and one, SCC-15, contained high levels of the EGF receptor as determined by immunoprecipitation via an EGF receptor-specific polyclonal antibody. Using a cloned EGF receptor complementary DNA as a probe, the level of EGF receptor RNA was found to be elevated four-fold in SCC-15 relative to normal cultured keratinocytes. When the same probe was used to identify EGF receptor gene fragments on a genomic DNA blot, the SCC-15 cell line was shown to possess an EGF receptor gene copy number amplified four to five times. Gene amplification results in the enhancement in the level of the EGF receptor in several carcinomas and could be responsible for the appearance of the transformed phenotype in these cells. PMID- 2984255 TI - Increased angiotensin-converting enzyme in peripheral blood monocytes from patients with sarcoidosis. AB - Angiotensin-converting enzyme (ACE) activity was measured in isolated peripheral blood monocytes and culture medium from 28 patients with sarcoidosis and compared with values obtained from monocytes of 25 normal control subjects. ACE activity was determined by radioimmunoassay of angiotensin II produced from angiotensin I. While there was no measurable ACE activity in monocytes or culture medium from normal controls under the conditions of our study, monocytes from patients with sarcoidosis all showed activity both in cells and culture medium. The mean ACE activity of monocytes from patients with sarcoidosis was 2.0 pg angiotensin II formed/min per 10(5) cells, and that released into medium over a 24-h interval was 30.4 pg angiotensin II/min per 10(5) cells. The monocyte ACE from patients with sarcoidosis was activated by chloride ions and inhibited by EDTA, captopril, and rabbit antiserum to purified human plasma ACE, indicating that enzymatic activity was effected specifically by ACE. Thus, our studies show a significant elevation and release of ACE by peripheral blood monocytes of patients with sarcoidosis under conditions where monocytes of normal control subjects do not demonstrate ACE activity. PMID- 2984257 TI - Morphology and laminar distribution of electrophysiologically identified cells in the pigeon's optic tectum: an intracellular study. AB - The responses of 65 cells to electrical stimulation of the contralateral optic nerve were intracellularly recorded in the pigeon optic tectum by using micropipettes filled with a solution of horseradish peroxidase. Nineteen of them were successfully labeled. Microscopic examination of the filled cells shows that our sample includes six pyramidal, ten ganglion, two stellate, and one bipolar horizontal cells. Thus, pyramidal and ganglion neurons constitute the most numerous types of cells in our sample. Pyramidal cells were located in layer II but mostly in its non-retinorecipient part, and they had restricted ascending dendritic trees oriented orthogonal to the tectal lamination. Ganglion cells were located in layer III with one exception, which was in sublayer IIi. These cells had non-oriented dendritic trees which ramify over considerable distances. Terminal dendritic branches from a number of pyramidal and ganglion cells extended superficially well within the region of optic fibers termination. In our study, ganglion cells constituted the efferent tectal elements. Pyramidal cells responded to optic nerve stimulation with a pure EPSP, with an EPSP-IPSP sequence, or with a pure IPSP. Ganglion cells always exhibited an IPSP either alone or preceded by an EPSP. Stellate and bipolar cells responded with a pure EPSP. The study of the laminar distribution of labeled and non-labeled cells shows from surface to depth, a gradual increase in the number of cells responding with an EPSP-IPSP or with a pure IPSP and a gradual decrease in the number of those exhibiting a pure EPSP. The analysis of the sensitivity of EPSPs and IPSPs to high frequency optic nerve stimulation shows that monosynaptic as well as polysynaptic EPSPs can be recorded from cells in the non-retinorecipient tectal region, a number of ganglion and pyramidal cells receive a direct retinal excitatory input as their dendrites pass through the region of optic endings, most IPSPs are polysynaptic, some cells located in the retinorecipient region may receive direct retinal inhibitory connections. PMID- 2984256 TI - Human thrombomodulin is not an efficient inhibitor of the procoagulant activity of thrombin. AB - The effect of human thrombomodulin isolated from placenta on the procoagulant activity of thrombin was studied and compared to that of rabbit thrombomodulin. The isolated protein was proved to be thrombomodulin because a rabbit antibody against the isolated protein blocked protein C activation by thrombomodulin in solution and also blocked the protein-C-activating cofactor activity of human umbilical vein endothelial cells. The affinity of human thrombomodulin for human thrombin in the presence of fibrinogen is 30 times less than that of rabbit thrombomodulin. This value is based on the measurements of the clotting time of human fibrinogen and thrombin in the presence of increasing amounts of thrombomodulin. Human thrombomodulin was also much less effective compared with rabbit thrombomodulin in inhibiting thrombin-induced human coagulation factor V activation. The ability to inhibit release of [3H]serotonin from washed human platelets was at least 10 times less using human thrombomodulin compared with rabbit thrombomodulin. A partially purified preparation of human lung thrombomodulin was also relatively ineffective in inhibiting thrombin-induced serotonin release from platelets, indicating that the difference between rabbit and human thrombomodulin is one of species rather than of tissue. Thus, while human thrombomodulin is a potent cofactor in protein C activation, it is not an efficient inhibitor of the procoagulant actions of thrombin. PMID- 2984258 TI - A combined Golgi and autoradiographic study of (3H)glycine-accumulating amacrine cells in the cat retina. AB - Golgi techniques were combined with electron microscopic autoradiography to identify four subpopulations of amacrine cell in the cat retina which accumulate (3H)glycine. These subpopulations include types A3, A4, A7(AII), and A8 amacrine cells. All are narrow-field cells with dendritic spreads of less than 100 micron. Quantification of silver grains showed that each subpopulation exhibits a consistent affinity for (3H)glycine. Type A8 cells were found to have the greatest affinity with normalized grain densities of 0.88-1.0 grains/micron 2 on a scale in which 1.0 represents the most heavily labeled cell. Type A4 cells were moderately labeled with grain densities ranging from 0.40 to 0.68 grains/micron 2. A7(AII) and A3 amacrines were lightly labeled with grain densities of 0.33 0.35 grains/micron 2 and 0.28-0.30 grains/micron 2, respectively. PMID- 2984259 TI - Nodular hidradenoma and hidradenocarcinoma. A 10-year review. AB - We studied eighty-nine nodular hidradenomas (NHs) and six clear cell hidradenocarcinomas (CCHs) reported in a 10-year period. NHs were more commonly seen in women (ratio of 1.7 to 1); the average age was 37.2 years, and they were located mainly on the head. One CCH had widely disseminated metastasis that led to death. Other CCHs underwent spontaneous regression. PMID- 2984260 TI - Alpha 1-antitrypsin and lysozyme in fibrous papules and angiofibromas. AB - The large stellate and polygonal cells observed in eleven fibrous papules and two angiofibromas were examined immunohistochemically for alpha 1-antitrypsin and lysozyme. The positive findings suggest that these cells are related to histiocytes rather than nevomelanocytes. PMID- 2984261 TI - Cyclic AMP phosphodiesterase activities in growing cells of baker's yeast (Saccharomyces cerevisiae). AB - The activities of two cyclic AMP phosphodiesterases of baker's yeast (Saccharomyces cerevisiae) were measured during diauxic batch growth on 2% glucose. The specific activity (units/mg of yeast protein) of the Mg-independent, high Km phosphodiesterase increased 20-fold throughout the 108 h cultivation. The specific activity of the Mg-dependent, low Km phosphodiesterase about doubled during glucose utilization and fell back to the initial level as the cells entered stationary phase. PMID- 2984262 TI - Adenylate cyclase activity as a function of forskolin concentration. AB - The activation of adenylate cyclase by forskolin was investigated in terms of the dissociation model of guanyl nucleotide binding protein (Ns). It was demonstrated that the biphasic forskolin concentration-response of adenylate cyclase could be explained by the dissociation of the beta subunit. The equations developed from such a theoretical approach gave an accurate description of concentration response data from S49 cultured mouse lymphoma cells. PMID- 2984263 TI - A glutathione adduct of prostaglandin A1 acts intracellularly to elevate cyclic AMP by inhibiting its extrusion. AB - Prostaglandin A1 inhibits the energy-dependent export of cyclic AMP from avian red cells. When exposed to PGA1, avian red cells rapidly accumulate the prostaglandin and convert it to a polar metabolite. Inhibition of cyclic AMP efflux correlates with the intracellular content of this metabolite. Data in the literature and our chemical and chromatographic analyses indicate that the prostaglandin metabolite is a glutathione adduct of PGA1. Thus, we hypothesize that a GSH adduct of PGA1 acts intracellularly to elevate cyclic AMP by blocking its extrusion. PMID- 2984264 TI - cAMP-dependent protein kinases of rat pituitary GH3 cells. AB - Two isoenzymes (type I and type II) of cAMP-dependent protein kinases were found in GH3 cytosol, isozyme type II activity being the predominant form (approximately 90%). Photoaffinity labeling of GH3 cell extracts with 8-N3 [32P]cAMP revealed three cAMP-binding proteins exhibiting molecular weights of 53,000, 51,000 and 48,000, respectively. The latter represents the regulatory subunit of type I isoenzyme whereas the 53,000 and 51,000 cAMP-binding proteins correspond to two different molecular forms of the type II isoenzyme regulatory subunit which are phosphorylated by a cAMP-dependent mechanism. PMID- 2984265 TI - beta-Adrenergic receptors and adenylate cyclase activity in murine lymphoid cell lines. AB - beta-Adrenergic receptors were determined in sixteen cultured tumor cell lines of T- and B-lymphoid derivation by the binding of the hydrophilic ligand [3H]CGP 12177. The number of receptors per cell varied from less than a hundred to over three thousand. Intracellular cAMP responses to isoproterenol and forskolin were measured for seven cell lines possessing a range of receptor numbers. No direct relationship was found between the isoproterenol-induced elevation of cAMP and the receptor number, and forskolin-induced increases in cAMP varied by more than 100-fold between cell lines. Adenylate cyclase activity was also measured in membrane preparations from the seven cell lines. The T-lymphoma line WEHI-22, the line with the highest receptor number (3700 per cell), showed very little response to any stimulatory agent. With the other four cell lines possessing receptors, the membrane-bound adenylate cyclase activity evoked by isoproterenol relative to that evoked by forskolin was found to be proportional to the number of receptors per cell. The number of receptors thus seems to determine the proportion of the inherent adenylate cyclase that can be activated by a beta adrenergic agent. The levels of adenylate cyclase induced in membranes by GppNHp and by fluoride, relative to those induced by forskolin, were substantially higher with some cell lines than with others. These data and the results from WEHI-22 cells indicate that lymphoid cell lines differ in their content of functional C and N proteins. PMID- 2984266 TI - Intracellular injection of cyclic GMP depresses cardiac slow action potentials. AB - Our laboratory has shown that intracellular injection of cyclic AMP (cAMP) transiently enhances slow APs in myocardial cells, presumably by phosphorylating slow channels. To test if cGMP also plays a role in cardiac slow channel function, superfusion with 8-Br-cGMP, and intracellular injections of cGMP were carried out in guinea pig papillary muscles (stimulated at 0.5 Hz at 37 degrees C). In normal (4.7 mM K+) Tyrode's solution, 0.1 mM 8-Br-cGMP depressed contractions and had variable effects on the duration of the fast APs. Slow APs were elicited by electrical stimulation (in 25 mM K+-Tyrode's solution) following the addition of 10 mM TEA and doubling the bath [Ca] (to 4.0 mM) or addition of 0.2 microM isoproterenol. Slow APs are dependent on the slow inward current carried through voltage- and time-dependent slow channels. 8-Br-cGMP (0.1 microM 1 mM) superfusion depressed or abolished slow APs and accompanying contractions. cGMP (5-100 mM Na+ salt in 0.2 M KC1) was injected by application of pressure pulses (40-75 psi, 1-30 sec duration) to the recording microelectrode. cGMP injection transiently depressed (n = 15) or abolished (n = 4) the slow APs. The effect began 1 min after the onset of the pulse, reached a maximum at 2 min and recovered fully within 5-6 min. Thus, it appears that the intracellular cGMP level can modulate the slow inward current in a direction opposite to that of cAMP. These effects may both be due to cyclic nucleotide-dependent phosphorylations. PMID- 2984267 TI - Epinephrine-induced sequestration of the beta-adrenergic receptor in cultured S49 WT and cyc- lymphoma cells. AB - Pretreatment of either intact wild type S49 lymphoma cells (WT) or the uncoupled variants, cyc-, H21a, or UNC with epinephrine results in the redistribution of 20 30% of the beta-adrenergic receptors into a light vesicle fraction in sucrose gradients. Since the variants are uncoupled with respect to hormonal stimulation of adenylate cyclase, it appears that productive interaction with Gs is not required for the sequestration of beta-adrenergic receptors. Characterization of the epinephrine-induced redistribution of the beta-adrenergic receptor has revealed the following: The EC50 for the redistribution in WT cells was between 100 and 200 nM. Pretreatment of WT cells with 50 nM epinephrine for 30 min induced only a slight redistribution of receptors in sucrose gradients but produced a significant desensitization of adenylate cyclase. The desensitization was characterized by an increase in the Kact of epinephrine stimulation of adenylate cyclase while the Vmax was unaltered. Pretreatment with 10 microM epinephrine resulted in a significant decrease in the Vmax (50%) of epinephrine stimulation of adenylate cyclase and a 3-fold increase in Kact in the heavy vesicles. The beta-receptors in the light vesicle fraction of WT were uncoupled from adenylate cyclase and displayed low affinity for epinephrine binding, comparable to the cyc-. The "desensitized" receptor in the light vesicle fractions of cyc- was capable of stimulating adenylate following reconstitution with cholate extracts of WT membranes containing Gs. The molecular weight of the photolabeled beta-receptor in the light vesicle fractions (65,000 +/- 2,000) was not significantly different from the Mr 65,000 polypeptide photolabeled in the heavy fractions. The Mr 55,000 beta-receptor polypeptide was not detected in the light vesicles. Our results suggest first that the redistribution of the beta receptor into light vesicles may follow an earlier stage of desensitization, and second that the beta-receptor in light vesicles while sequestered from Gs is capable of activating adenylate cyclase. PMID- 2984268 TI - Dietary sodium bicarbonate for high-producing Holstein cows over complete lactations. AB - Several concentrates with sodium bicarbonate (experimental) were compared with the same concentrates without sodium bicarbonate (control) throughout two lactations. During first lactation, two control concentrates were used in sequence in one comparison and three in another. Toward the end of first lactation, sodium bicarbonate was increased from 1.5 to 2.5% in concentrates, and that concentration was continued throughout the second lactation. During second lactation high-moisture corn with soybean meal was one concentrate; the other included half high-moisture barley and half dry corn and soybean meal. Fifty-two and 60 lactations of cows fed control and experimental rations contributed feed intake and production data. Ninety-four and 95 lactations of cows fed control or experimental rations contributed 305-day, mature-equivalent production data over three lactations. Inclusion of sodium bicarbonate in these rations had little effect on feed consumption, milk production, fat content of milk, efficiency of milk production, or change of body weight. The kind of concentrate did not alter the effect of sodium bicarbonate. Cows consuming control rations produced 8898 kg milk with 312 kg fat (305-day mature equivalent). Cows using bicarbonate produced 8972 kg milk with 312 kg fat. PMID- 2984269 TI - Effect of adrenocorticotropic hormone and associated hormonal responses on semen quality and sperm output of bulls. AB - The objective was to determine if semen quality and output could be affected by pharmacological doses of adrenocorticotropin. Three Holstein bulls, one 7 yr old and two yearlings, were treated with 200 IU adrenocorticotropic hormone every 8 h for 6 days. Effects of treatment on semen traits and peripheral concentrations of glucocorticoids, testosterone, and androstenedione were measured. Viability of spermatozoa (percentage motility and percentage intact acrosomes), ejaculate volume, sperm concentration, and weekly sperm output were unaffected by adrenocorticotropin treatment up to 8 wk posttreatment. The proportion of spermatozoa with cytoplasmic droplets and head abnormalities was elevated slightly from .8 +/- .1 and 4.2 +/- .3% to 1.3 +/- .2 and 5.8 +/- .4% during treatment. Total glucocorticoids increased from 11.9 +/- 2.7 ng/ml before treatment to 73.5 +/- 4.1 ng/ml during treatment. Testosterone decreased in the yearling bulls from 5.5 +/- .9 ng/ml plasma before treatment to .5 +/- .5 ng/ml afterward; the decrease began 8 h following the initial adrenocorticotropin injection and persisted until 24 h following last injection. The mature bull had normal testosterone concentrations for the first 4 days of injection, and decreased concentrations for the last 2 days of injection. Semen viability, concentration, and sperm output are unaffected by a pharmocological administration of adrenocorticotropin and subsequent marked increase of glucocorticoids and decrease of testosterone. Only a small increase of semen content of immature sperm or sperm with abnormal heads may be associated with these marked endocrine changes. PMID- 2984270 TI - Enteric viral infections of calves and passive immunity. AB - At least eight viruses have been identified, four within the last 5 yr, that produce diarrhea and pathological intestinal lesions in experimentally inoculated calves. Coronavirus and rotavirus frequently are associated with the neonatal calf diarrhea syndrome, but the etiologic role of the newly identified viruses is undefined. All diarrheal viruses replicate within small intestinal epithelial cells, resulting in variable degrees of villous atrophy. Immunity against these viral infections, therefore, must be directed toward protection of the susceptible intestinal epithelial cells. Because most of these viral infections occur in calves less than 3 wk of age, passive lactogenic immunity within the gut lumen plays an important role in protection. This report reviews methods of boosting rotavirus antibody responses in bovine mammary secretions and analyses of passive and active immunity in calves supplemented with colostrum and challenged by rotavirus. Results indicate rotavirus immunoglobulin G1 antibodies in colostrum and milk were elevated after intramuscular and intramammary vaccination of pregnant cows with an Ohio Agricultural Research and Development Center rotavirus vaccine but not after intramuscular immunization with a commercial rota-coronavirus vaccine. Feeding colostrum from intramuscular plus intramammary immunized cows to newborn calves challenged by rotavirus prevented diarrhea and shedding of rotavirus. PMID- 2984271 TI - Effect of forage: concentrate on kinetics of forage fiber digestion in vivo. AB - With five rumen-fistulated Holstein steers in a Latin square design, we determined the effect of dietary concentrate (0, 20, 40, 60, or 80% cracked corn) on kinetic characteristics influencing forage fiber digestion in vivo. Rate and potential extent of neutral detergent fiber degradation were determined for fescue hay in situ by nylon bag technique. Rate of fiber passage from the rumen was measured by fecal excretion of chromium-mordanted fescue cell walls. Apparent extent of forage fiber digestion was predicted by a model in which fiber disappearance from the rumen is conceptualized as the sum of two competing first order processes, digestion and passage, modified further by a discrete lag time during which fiber passes from the rumen before digestion commences. Stepwise multiple regression analysis revealed that potential extent of fiber degradation in situ was the primary determinant of depression of fiber digestibility as dietary concentrate increased. Rate of digestion, rate of passage, and lag effects collectively accounted for a small portion of the depression of fiber digestibility. PMID- 2984272 TI - Vasoactive intestinal peptide and bipolar affective illness. Evidence for an effect of lithium. AB - In an attempt to evaluate the role of VIP in affective disorder, measurements of lymphocyte VIP receptors, and plasma and CSF VIP levels were made in unmedicated and lithium-treated euthymic bipolars and controls. Lithium decreased plasma (P = 0.01) and CSF (P = 0.05) VIP levels and increased the affinity (decreased the KD) of the VIP lymphocyte receptor (P less than 0.01). This effect may be relevant to the psychotropic action of lithium in manic-depressive illness. PMID- 2984273 TI - Reversal of vanadate-induced inhibition of Na-K ATPase. A possible explanation of the therapeutic effect of carbamazepine in affective illness. AB - There is evidence that carbamazepine is of therapeutic benefit in manic depressive illness. There is also evidence that raised vanadium levels may be of aetiological importance in manic depressive illness. The present study examined the effect, in vitro, of therapeutic concentrations of carbamazepine on the inhibition by ammonium metavanadate of the Na-K ATPase of erythrocytes from normal and from manic depressive subjects. The inhibition by vanadate was largely reversed by carbamazepine. This effect may be related to the therapeutic action of carbamazepine in manic depressive illness. PMID- 2984274 TI - Preservation of the alveolar ridge with hydroxylapatite tooth root substitutes. AB - A total of 49 patients, who were followed for 3 to 31 months, have had 92 hydroxylapatite root implants placed in fresh extraction sockets without soft tissue closure. Hydroxylapatite root implants maintain approximately twice as much alveolar bone as the contralateral control sites without implants. It is believed that this method is a more effective and efficient procedure to preserve alveolar bone for retention of dentures than previously described procedures. PMID- 2984275 TI - Nutrition evaluation of published weight-reducing diets. AB - The objective of this study was to determine the nutritional adequacy of some of the popular published diet plans. Diet analyses were made using the University of Massachusetts Nutrient Data Bank. Not one of the 11 diets evaluated provided 100% of the U.S. Recommended Daily Allowances for the 13 vitamins and minerals studied. The nutrients most often below recommended levels were thiamin, vitamin B-6, vitamin B-12, calcium, iron, zinc, and magnesium. Vitamin and mineral supplementation may be warranted for individuals following some diet plans. PMID- 2984276 TI - Reticular formation of the lower brainstem. A common system for cardio respiratory and somatomotor functions. Cross-correlation analysis of discharge patterns of neighbouring neurones. AB - Temporal relations of discharges of 73 pairs of neurones located in the medial parts of the reticular formation of the lower brainstem were studied by cross correlation analyses in chloralose-urethane anaesthetized dogs. The action potentials of 2 or 3 neighbouring neurones were recorded with one electrode simultaneously. Uncorrelated discharges of neurones and 4 different types of correlated discharges were observed in cross correlation histograms: they were: (1) rhythmic couplings with frequencies between 2 and 5 Hz related to the same rhythm in the EEG; (2) strong, non-rhythmic couplings with short latencies up to 5 ms; (3) a combination of strong and rhythmic couplings, and (4) high-frequency oscillation couplings. Most pairs of neurones showed different types of correlation during the recordings. The different forms of correlated discharge behaviour could be related to different types of functional organization of the neuronal network in the reticular formation. PMID- 2984278 TI - Multiple recurrence of digital pigmented villonodular tenosynovitis: a case report. AB - The propensity for digital pigmented villonodular tenosynovitis to recur is well known. A patient who has had 11 consecutive recurrences of the tumor over a 14 year period has retained a functional finger. PMID- 2984277 TI - Inhibitory effects on steroid production from isolated adrenal cells of rhesus monkey (Macaca mulatta) of pro-opiomelanocorticotrophic peptides. AB - Adrenal glands from Rhesus monkeys (Macaca mulatta) of 160 days gestation, newborn, 2 months-old infants or 6 months-old infants were excised and prepared, by a collagenase digestion, as a cell suspension. The cells were incubated with 10 pg/ml, 100 pg/ml or 1 ng/ml of a peptide of the ACTH/pro-opiomelanocortin 'family', 57K, 31K, 20K, alpha MSH, ovine-CLIP or gamma LPH either in the presence or absence of 166 pg/ml ACTH1-39. The production by cortisol and androstenedione was measured by radioimmunoassay. Using the steroid production by aliquots of the cell suspension with either no stimulating agent or ACTH1-39 alone as controls, the net influence of these different peptides on basal or ACTH1-39-stimulated production was observed. alpha MSH, ovine-CLIP and gamma LPH had no influence on either basal or stimulated cortisol or androstenedione production. Corticotrophic peptides of 57K, and 20K and pro-opiomelanocortin each had a steroidogenic activity alone, in all age groups. In the fetal and newborn monkeys' adrenal cells, peptides of 57K and 20K at 1 ng/ml had an inhibitory influence on ACTH1-39 stimulated cortisol and androstenedione production. The influence of the 20K peptide is partially inhibitory as the steroidogenic potential of this peptide is not additive with that of ACTH1-39. These results show that, as observed in other species, that the ACTH/pro-opiomelanocortin range of peptides are inhibitory to the action of ACTH1-39 in the developing adrenal. PMID- 2984279 TI - Isolation of stable mouse L cell transfectant expressing human T cell differentiation antigen, Tp40. PMID- 2984280 TI - BLAST-2 [EBVCS], an early cell surface marker of human B cell activation, is superinduced by Epstein Barr virus. AB - Activation of human B cells by pokeweed mitogen (PWM), protein A, anti-IgM, or EBV infection results in the expression of a new surface antigen, termed BLAST-2 [EBVCS]. This marker appears before the cells undergo blast transformation as assessed by the initiation of DNA synthesis and expression of the BLAST-1 antigen. Thus, the BLAST-2 [EBVCS] antigen is expressed on both activated and lymphoblastoid cells. The antigen is, in addition, restricted to B cells, as it is not found on cells of T or myeloid lineage derived from peripheral blood, cell lines, or neoplastic cells. However, it is readily detected on chronic lymphocytic leukemia cells of B cell origin and in the germinal centers of tonsils and lymph nodes. Like the BLAST-1 antigen, BLAST-2 [EBVCS] is expressed at a high level only on EBV-transformed B lymphoblasts and has a m.w. close to 45,000. Immunoprecipitation experiments show, however, that the two antigens are expressed on distinct populations of molecules. PMID- 2984281 TI - Prolactin receptors on human T and B lymphocytes: antagonism of prolactin binding by cyclosporine. AB - Prolactin (PRL) receptors have been identified recently on human peripheral blood mononuclear cells (MNC) and may be involved in the regulation of cell-mediated immunity. Cyclosporine (CsA), an immunosuppressive cyclic endecapeptide utilized to prolong graft survival in human organ transplant patients, affects PRL binding to MNC. At concentrations of CsA from 10(-10) through 10(-8) M, the amount of PRL bound to MNC markedly increased to ca. 400% of controls, whereas CsA concentrations of 10(-6) and 10(-5) M totally inhibited PRL binding to lymphocytes. The ability of low concentrations of CsA to enhance PRL binding was temperature-dependent and did not occur when binding assays were conducted at 4 degrees C. PRL displaced [3H]CsA from lymphocytes with ca. 50% displacement at 10(-9) M PRL and total displacement at concentrations of 10(-7), 10(-6), and 10( 5) M. Growth hormone did not displace [3H]CsA in similar experiments. CsA also did not alter the binding of a beta-receptor antagonist to MNC, again suggesting that CsA was specific in its antagonism of PRL binding. A CsA analog with no immunosuppressive action, cyclosporin H, did not alter PRL binding to MNC. Furthermore, PRL receptors were demonstrated on four cell lines of human and mouse origin. Finally, PRL receptors were identified on purified populations of T and B lymphocytes isolated from human spleens, and CsA again inhibited PRL binding at concentrations of 10(-7) and 10(-6) M. The presence of PRL receptors on T and B lymphocytes suggests that PRL may be involved in the regulation of humoral and cell-mediated immunity, and that one effect of CsA on immune function may be its ability to inhibit the effects of PRL action on these lymphocytes. PMID- 2984282 TI - Characteristic T cell dysfunction in patients with chronic active Epstein-Barr virus infection (chronic infectious mononucleosis). AB - We evaluated immune functions in 16 patients with chronic active Epstein-Barr virus (EBV) infection (chronic infectious mononucleosis). Chronic infectious mononucleosis is an illness characterized primarily by chronic and occasionally disabling fatigue and other constitutional complaints, only sometimes beginning with an episode of acute infectious mononucleosis, and associated with an abnormal pattern of serum antibodies to EBV. In these patients, the frequency of circulating EBV-infected B cells that manifested spontaneous outgrowth in vitro was comparable to that found in EBV-seropositive normals, and the levels of EBV specific suppressor activity were also normal. Upon stimulation with polyclonal activators, unseparated cells from these patients produced a relatively normal number of immunoglobulin-secreting cells. However, when purified T cells from these patients were mixed with normal mononuclear cells in co-culture, immunoglobulin production was strikingly suppressed. The degree of this T cell suppression correlated directly with the abnormally elevated titer of antibody to the early antigens of EBV. Interestingly, during normal convalescence from acute EBV-induced infectious mononucleosis a period is also seen during which T cells suppress the response of allogeneic but not autologous cells. Thus, from an immunologic viewpoint, patients with chronic active EBV infection appear "frozen" in a state typically found only briefly during the convalescence from acute EBV infection. PMID- 2984283 TI - Generation of activated killer (AK) cells by recombinant interleukin 2 (rIL 2) in collaboration with interferon-gamma (IFN-gamma). AB - Activation of human peripheral blood mononuclear cells (PBMC) by interleukin 2 (IL 2) and the role of interferon-gamma (IFN-gamma) in the IL 2-induced activation were investigated. Activated killer (AK) cells against NK-resistant tumor cell lines were induced in the medium containing recombinant IL 2 (rIL 2) and autologous serum without any other stimulating agents. AK activity was induced by doses of rIL 2 as low as 3 U/ml, and reached a maximum at 10(3) U/ml. Incubation of PBMC with rIL 2 resulted in IFN-gamma production and augmented NK activity after 1 day of culture, and in induction of AK cells and proliferative response after 2 days of culture. These results suggested that endogenous IFN gamma was required for rIL 2-induction of AK cells and proliferative response. To prove this, PBMC were cultured with rIL 2 and rIFN-gamma or were pretreated with rIFN-gamma before culture with rIL 2. Both rIFN-gamma treatments of PBMC augmented rIL 2-induced AK activity and proliferative response. rIL 2-induced IFN gamma production was also enhanced by the rIFN-gamma pretreatment of PBMC. The addition of anti-IFN-gamma antibody to rIL 2 cultures abrogated the rIL 2-induced NK augmentation, AK generation, and proliferative response in proportion to the decreased amounts of endogenous IFN-gamma detectable in culture. rIFN-gamma and/or rIL 2 cultures of PBMC increased Tac antigen expression on cell surfaces as measured by flow cytometry. Enhanced Tac expression by rIL 2 was abrogated by adding anti-IFN-gamma antibody. These data indicate that: 1) AK generation and IFN-gamma production are mediated by IL 2, and 2) IFN-gamma production may be required for IL 2 induction of AK cells and proliferative response. These finding are consistent with the hypothesis that AK generation involves a collaboration between IL 2 and IFN-gamma, in which IL 2 stimulates PBMC to produce IFN-gamma, which in turn acts as a differentiation signal that may be involved in the IL 2 initiated AK generation and proliferative response. PMID- 2984284 TI - Lyme disease spirochetes induce human and murine interleukin 1 production. AB - IL 1 is a major immunoregulatory molecule produced by macrophages, and it appears to be the molecular orchestrator of nonspecific host defense mechanisms against a variety of environmental insults. Many investigators have used artificial agents to stimulate macrophages to produce IL 1. We now report production of large quantities of IL 1 after a physiologic stimulus. The Lyme disease spirochete, recently isolated and adapted for growth in vitro, was used to stimulate P388D1 cells or human peripheral blood monocytes. Spirochetes were added to confluent macrophage cultures in serum-free RPMI at a ratio of 10:1. The release of IL 1 was dose-dependent. The 24-hr supernatant IL 1 activity was determined by using the thymocyte Con A co-mitogenesis assay. Activity was not due to an endotoxin on, or produced by, the spirochete. A polymyxin B affinity column failed to remove activity, and polymyxin B in the spirochete-macrophage culture had no effect on IL 1 production. Supernatants were collected, were concentrated, and were subjected to size exclusion HPLC. Three areas of activity were found in P388D1 cell supernatants (Mr greater than 60,000, 40,000, and 20,000), whereas two peaks (Mr 23,000 and 13,000) were found in human monocyte supernatants. The Mr 20,000 and 13,000 peaks from murine and human cell supernatants, respectively, were subjected to SDS-PAGE and were shown to be single bands (Mr 12,400 for the mouse IL 1 and Mr 13,500 for the human IL 1). Isoelectric focusing of column purified IL 1 preparations showed two different pI in both human (pI 7.25 and 4.4 to 5) and murine (pI 7.25 and 5.55) IL 1. Fibroblasts cultured with murine or human IL 1 preparations demonstrated both an increase in secreted collagenase and increased cell proliferation. Thus, a physiologic stimulus and simple biochemical techniques produce large amounts of very pure mouse or human IL 1. That this IL 1 is produced by Lyme disease spirochete-stimulated macrophages may explain some of the clinical manifestations of Lyme disease. PMID- 2984285 TI - Polymorphisms within the HLA-DRw6 haplotype. I. Restriction fragment length variation and its correlation with serology. AB - The Dw6/DRw6 complex, one of the MHC class II specificities that can be defined by cellular techniques and by serology, probably has one or more immunoregulatory functions. To obtain information on the molecular structure of the DRw6 region, we studied several DRw6 homozygous cell lines, of which three were of consanguineous origin. DNA-DNA hybridization comprised the use of seven restriction enzymes in combination with three DR beta cDNA probes. The obtained results were compared with similar analyses of an HLA homozygous cell panel, expressing DR1-w8 specificities. This comparison indicated that in DRw6 homozygous individuals the coding potential for DR beta chains resembles closely that of all other DR specificities, thus identifying DRw6 as a regular DR region. In addition, we found a restriction fragment length pattern unique for DRw6, indicating the possibility to type for DRw6 by DNA-DNA hybridization. Comparisons within the DRw6 cell panel revealed the occurrence of several HLA class II DNA subtypes. These subdivisions partly correlated with serologically obtained reaction patterns. No correlation, however, could be observed between the different DNA subtypes and cellular reaction patterns as obtained by MLC and T cell cytotoxicity. PMID- 2984286 TI - Membrane complement receptor type three (CR3) has lectin-like properties analogous to bovine conglutinin as functions as a receptor for zymosan and rabbit erythrocytes as well as a receptor for iC3b. AB - Human leukocyte complement receptor type three (CR3) was shown to be lectin-like and to resemble bovine serum conglutinin (K) in that it bound to both iC3b and unopsonized yeast (Saccharomyces cerevisiae), and was inhibited by EDTA or N acetyl-D-glucosamine (NADG). CR3 and K also bound to zymosan (Z), a yeast cell wall extract that contains primarily polysaccharide and no detectable protein. However, structural differences and the absence of K on bovine phagocytes indicated that CR3 was not the human homologue of bovine K. Phagocytic and respiratory responses to unopsonized Z were CR3 dependent because they were inhibited by monoclonal antibodies specific for the alpha-chain of CR3 and did not occur with phagocytes from patients with a genetic deficiency of CR3. The binding of CR3 to Z did not require opsonization of the Z with neutrophil secreted C3, as Z binding and responses were not inhibited by Fab anti-C3. In addition, CR3-dependent binding of yeast occurred with neutrophils from which protein secretion was blocked by fixation with paraformaldehyde. Rabbit erythrocytes (RaE) also bound weakly to neutrophil CR3 and triggered ingestion. Anti-CR3 not only blocked the binding and ingestion of RaE but also blocked selectively the ingestion of RaEC3b without affecting the strong binding mediated by CR1. Even though sheep E and sheep EC3b were not ingested by neutrophils, a weak binding of CR3 to sheep E was suggested by the finding of 20 to 40% inhibition of sheep EAIgG ingestion by anti-CR3. Such inhibition was only observed in buffers that allowed activity of the CR3 binding site and not in buffers containing either EDTA or NADG. An apparently contradictory finding was that the weak CR3-dependent binding of Z triggered neutrophil ingestion and a superoxide burst, whereas the avid CR3-dependent binding of sheep EC3bi did not induce significant ingestion or a respiratory burst. Blocking studies with monoclonal antibodies specific for different epitopes of the alpha-chain of CR3 suggested that this might result from the presence of two distinct binding sites in CR3: one site for fixed iC3b that did not trigger functions, and a second function-triggering site for Z that did not bind to fixed iC3b. PMID- 2984287 TI - Studies of leukotriene B4-specific binding and function in rat polymorphonuclear leukocytes: absence of a chemotactic response. AB - Rat PMN isolated from peripheral blood show a small amount of high-affinity (specific) binding of [3H]-LTB4 at nanomolar concentrations. This binding is reversible and has a stereospecificity similar to rat PMN aggregation in response to several LTB4 analogs. This population of binding sites shares many characteristics with a population of high-affinity binding sites in human PMN; however, human PMN bind a significantly greater amount of [3H]-LTB4 to a second population of specific binding sites that is not present in rat PMN. The aggregation responses of human and rat peripheral blood PMN to LTB4 are similar in magnitude and specificity, but unlike human PMN, LTB4 fails to elicit a chemotactic response in rat PMN at concentrations from 10(-10) M to 10(-6) M. Rat PMN also fail to metabolize exogenous LTB4 when compared with human PMN. These data suggest that different PMN functions, such as chemotaxis and aggregation, may involve different classes of specific receptors. The finding that rat PMN do not exhibit chemotaxis to LTB4 calls for a reevaluation of the relevance to inflammation in humans of studies of inflammation performed in rat models. PMID- 2984288 TI - D-Penicillamine: analysis of the mechanism of copper-catalyzed hydrogen peroxide generation. AB - Recent studies have suggested that the inhibition of lymphocyte mitogenesis by D penicillamine in the presence of copper could be mediated by the formation and action of hydrogen peroxide. To explore this possibility further, we first sought evidence of H2O2 generation by D-penicillamine in a cell-free system by a) measurement of copper-catalyzed D-penicillamine oxidation and the requirement for oxygen in this process; b) direct measurement of H2O2 formation during D penicillamine oxidation by the peroxidase-mediated oxidation of fluorescent scopoletin; and c) evaluation of the possible synthesis of O2- during D penicillamine oxidation. The addition of copper to D-penicillamine in physiologic buffer catalyzed D-penicillamine oxidation in a dose-dependent fashion. D penicillamine oxidation was accompanied by O2 consumption with a molar ratio of approximately 2:1, but did not occur under anaerobic conditions. Furthermore, D penicillamine oxidation resulted in the formation of amounts of H2O2 stoichiometrically equivalent to oxygen consumption (i.e., 1:1). Copper-catalyzed D-penicillamine oxidation caused reduction of nitroblue tetrazolium in a reaction blocked by superoxide dismutase, suggesting the formation of O2-. Additional studies confirmed that D-penicillamine inhibited PHA-induced mitogenesis of lymphocytes in the presence of copper, and that catalase protected the cells from this action. Furthermore, when polymorphonuclear leukocytes were incubated with D penicillamine plus copper, hexose monophosphate shunt activity increased up to threefold with abrogation of this stimulation by catalase. None of the effects of D-penicillamine plus copper on cells were diminished by hydroxyl radical scavengers mannitol or benzoate. These results are consistent with oxygen dependent copper-catalyzed oxidation of D-penicillamine in aqueous solutions leading to the formation of O2- and H2O2. H2O2 produced by this reaction can inhibit lymphocyte mitogenesis and stimulate neutrophil hexose monophosphate shunt activity in vitro and may be relevant to the therapeutic effects of D penicillamine in vivo. PMID- 2984290 TI - Polymorphism of kappa variable region (V kappa-1) genes in inbred mice: relationship to the Ig kappa-Ef2 serum light chain marker. AB - We describe here the cloning and complete sequence analysis of the rearranged kappa variable region gene from the V kappa-1A-producing myeloma (C.AL20-TEPC 105). A 5'-flanking region probe from the V kappa-1A gene has been used to study the V kappa-1 germ-line gene family in strains of mice differing at the Ig kappa Ef2 locus. All Ef2a strains examined possess an identical pair of BamHI restriction fragments strongly hybridizing to the 5' probe. Surprisingly, only two of the six strains of mice previously designated Ef2b (NZB and C58) possessed clearly altered restriction fragment sizes for V kappa-1 genes. The remaining four Ef2b strains, namely BDP/J, CE/J, I/LnJ, and P/J, appear to carry V kappa-1 genes similar to those of Ef2a strains. It is suggested that these strains may carry a third form of the V kappa-1A gene, differing in the protein coding region but indistinguishable at the DNA level with the use of BamHI or EcoRI. Alternatively, these strains may fail to express V kappa-1A light chains due to a regulatory defect involving this subgroup of kappa genes. PMID- 2984289 TI - Retrovirus-mediated immunosuppression. I. FeLV-UV and specific FeLV proteins alter T lymphocyte behavior by inducing hyporesponsiveness to lymphokines. AB - Murine splenocytes were used to study the in vitro immunosuppressive effects of UV-inactivated feline leukemia virus (FeLV-UV). FeLV-UV blocks both alloantigen (DBA/2)-induced and Con A-induced proliferation of C57BL/6 splenocytes in a dose dependent manner. Furthermore, C57BL/6 anti-DBA/2 mixed lymphocyte cultures containing FeLV-UV fail to develop detectable DBA/2-specific cytolytic activity, although FeLV-UV has no effect on the cytolytic activity of preformed C57BL/6 anti-DBA/2 cytolytic T cells (CTL). Disruption of lymphocyte proliferation and CTL generation by FeLV-UV could not be overcome by the addition of exogenous lymphokines. These data suggest that FeLV-UV can interfere with the lymphokine reactivity of alloactivated lymphocytes. In fact, FeLV-UV blocks the lymphokine induced proliferation of the murine IL 2-dependent cell line CTLL-20. The CTLL-20 cells were subsequently used to study the mechanism(s) by which retroviruses alter T lymphocyte function. Normally, CTLL-20 cells undergo significant proliferation when cultured in EL4 SN, an IL 2-containing culture supernatant from PMA-stimulated EL4 cells. This lymphokine-induced CTLL-20 proliferation is abrogated in a dose-dependent manner by UV-inactivated murine leukemia virus (MuLV-UV), FeLV-UV, and a purified 15,000 dalton viral protein, p15, derived from FeLV. Suppression of CTLL-20 proliferation requires only brief contact (6 hr) with FeLV-UV or with p15, but is most efficient after prolonged (24 hr) contact with these agents. Furthermore, suppression of CTLL-20 proliferation by FeLV-UV and p15 is reversible, because CTLL-20 cells which have been pretreated for 24 hr with FeLV-UV or p15 are equally as efficient at responding to EL4 SN as untreated CTLL-20. Additional studies indicate that CTLL-20 cells continue to remove IL 2 activity from EL4 SN in the presence of suppressive concentrations of FeLV-UV, and that suppressive concentrations of FeLV-UV do not remove IL 2 activity from EL4 SN. This suggests that FeLV does not block CTLL-20 proliferation by absorbing or inactivating IL 2, or by occluding IL 2 receptors, and that T lymphocytes develop an insensitivity to lymphokines after contact with FeLV-UV, which may be caused by a metabolic, rather than an immunologic, defect. Because lymphokines are requisite signals for T cell function, considerable immunosuppression would be associated with acquired lymphokine insensitivity. PMID- 2984291 TI - cDNA cloning of the murine transferrin receptor: sequence of trans-membrane and adjacent regions. AB - We previously purified the murine transferrin receptor from cultured myeloma cells and determined the amino acid sequence of six tryptic peptides. We now report the cloning of cDNA encoding the murine transferrin receptor. A tryptic peptide containing a region of six consecutive amino acids, all encoded by four or fewer codons, was used to design two overlapping oligonucleotide probes, 17 and 14 nucleotides in length. These probes were used to screen a lambda gt10 cDNA library from NS-1 myeloma cells. Of approximately 400,000 plaques screened, two hybridized strongly to both probes. A subfragment of one clone that hybridized with both oligonucleotide probes was found to encode the tryptic peptide from which the probes were derived, as well as another sequenced tryptic peptide. Comparison of the sequence with that of the human transferrin receptor shows a high degree of conservation of the sequences surrounding and penetrating the membrane, including cysteine residues that may be involved in interchain disulfide bonding and/or covalent attachment of lipid. The current data, when combined with the published sequence of the human receptor, allow assignment of all six tryptic peptides to a single chain, supporting the idea that the receptor is a homodimer. A 4.9-kb messenger RNA was found in several cultured murine and human tumor cell lines, but transferrin receptor messenger RNA was not detectable in murine spleen. An additional RNA species of 2.7 kb was present in approximately equal abundance in the murine myelomas NS-1 and C118 but was absent from T lymphomas TIKAUT, ST-1, and ST-4. PMID- 2984292 TI - Germline VH genes in an a3 rabbit not typical of any one VHa allotype. AB - We have undertaken investigations in the rabbit of VH genes that may be responsible for the observations of VHa allotypes unexpected from an animal's pedigree (latent allotypes). A short cDNA probe was prepared and shown to be specific for VHa2 mRNA. Southern analyses with short and large probes were unrevealing but screening of a lambda phage library from a VHa3-expressing animal identified a number of unusual genes. These VH genes are remarkable in that they are far closer to one another in the genome (in one case 3085 bps) than VH genes reported in mouse or man, they are highly homologous over long stretches of sequence, and they encode proteins not typical of any one VHa allotype. Proteins similar to the sort encoded by these genomic V-regions may explain some of the observations of latent allotype. Our data suggest that the allelic behavior of VHa allotypes is not due to allelism of a regulatory mechanism that acts upon identical VH genes in rabbits of different VHa phenotypes. PMID- 2984293 TI - Rapid microassays of phagocytosis, bacterial killing, superoxide and hydrogen peroxide production by human neutrophils in vitro. AB - Simple, rapid microassays for simultaneous measurement of phagocytosis, bacterial killing, superoxide and hydrogen peroxide production by human neutrophils in vitro are described. All assays employ 96-well flat bottom tissue culture plates which were incubated on a microtitre plate shaker at 37 degrees C. The separate evaluation of ingestion and intracellular killing of E. coli and S. aureus was based on the incorporation of [3H]uridine into viable extracellular bacteria. There was good correlation between plate counts of viable bacteria and amount of radiolabel incorporation. Phagocytosis and killing can be measured in a maximum of 100 microliter reaction mixture, requiring only 2.5 X 10(5) neutrophils per test and the assay is complete within 60 min. Assay of superoxide production by stimulated neutrophils was based on superoxide-dependent reduction of ferricytochrome c as measured spectrophotometrically at 550 nm in wells of tissue culture plates containing 150 microliter of reaction mixture. The assay requires only 1.25 X 10(5) neutrophils per test and is complete within 50 min. Quantitation of hydrogen peroxide was based on horseradish peroxidase-dependent oxidation of phenol red. The technique is as for superoxide detection except that the reaction must be terminated by the addition of 1 M NaOH at the desired time intervals. None of the assays require sampling during the incubation period. The principal advantages of the described techniques are increased simplicity and speed, requirement of low numbers of neutrophils and applicability to analysis of large number of samples in parallel. PMID- 2984294 TI - Nosocomial infection with hepatitis A. AB - Nosocomial spread of hepatitis A is very uncommon but may occur under unusual circumstances, as shown by the incident described here and by the few other published reports. In this incident it is concluded that the patient, who was the index case, was excreting hepatitis A virus in the faeces 16 days before jaundice developed and 17 days before alanine aminotransferase (ALT) values reached a peak. PMID- 2984295 TI - Prevalence of antibodies to hepatitis A virus among urban children aged 0-7 years in Yugoslavia. AB - In a study of the prevalence of hepatitis A virus antibody in urban Yugoslav children aged 0-7 years, 18.2% were positive; most of them were only a few months old. The maternal origin of antibody in these very young children could be inferred from the rapid decrease of antibody during the first 5-7 months of life, following which period until the age of 7 years, there was no significant increase in the proportion with antibody. It was concluded that the age range included in this study (0-7 years) does not represent a time of life when most urban Yugoslav children come into contact with hepatitis A virus. PMID- 2984296 TI - Influenza C virus in the United Kingdom. PMID- 2984297 TI - [Histopathological study of carcinoma of the cervix stage Ia]. AB - Seventy-three cases of microinvasive carcinoma of the cervix (restricted to Japanese criteria) were observed microscopically and compared with 67 cases of CIS. The lesion is more extended in stage Ia cases than those of CIS. There is a positive correlation between the depth of permeation and circular and longitudinal extensions of stage Ia cancer. Multiple invading lesions are observed in 52.9% of stage Ia cases. The lesion adjoining the microinvasive area is occupied by CIS in 50%. A tendency to differentiation is observed in 92.6% of Ia carcinoma, but in only 38.6% of CIS. We classified the invasive area into 3 grades: keratinizing type, large cell non-keratinizing type and small cell non keratinizing type. Incidences among 654 invasive areas were calculated to be 12.4%, 68.3% and 19.3% respectively. Round cell infiltration around the invasive area is observed in 81.3% of cases. Mitotic frequency is lowest in the microinvasive area and highest in CIS cases. Cancer cells with a prominent nucleolus are found more often in the microinvasive area, than CIS and CIS areas of Ia cases. Among 95 cases with stromal invasions not longer than 3mm, vessel permeations are found in 4(4.2%) and confluent invasions in 7(7.4%). PMID- 2984298 TI - [Immunoperoxidase demonstration of papillomavirus antigen in dysplasia of the uterine cervix]. AB - Recently, it has been widely recognized that human papillomavirus (HPV) infection plays an important role in the etiology of carcinoma of the cervix. Paraffin sections of cervical malignancies and premalignancies were screened for the presence of papillomavirus antigen by means of peroxidase-antiperoxidase (PAP) method. No case of invasive carcinoma, carcinoma in situ or normal cervix revealed positive HPV staining. In 81 cases (18%) of 461 cervical dysplasias, positive reaction for HPV was detected as brown intranuclear precipitates. The mean age of the women with HPV was significantly lower than that of women without HPV. Characteristic findings of histology of the dysplasias with HPV were koilocytosis and binucleation. PMID- 2984299 TI - [Acute onset of insulin-dependent diabetes mellitus caused by Epstein-Barr virus infection]. PMID- 2984302 TI - Differential ability of human blood monocyte subsets to release various cytokines. AB - We have shown that two human monocyte subsets can be isolated from the peripheral blood of healthy donors; these subsets possess different morphological, cytochemical, functional, and in vivo trafficking properties [1]. In this report, these two subsets were further characterized. One subset (intermediate monocytes, IM) has been shown to have significantly lower acid phosphatase activity and total cellular protein content as well as lower peroxidase activity when compared with another subset (regular monocytes, RM). The overall activation status of the two subsets (as determined by their alkaline phosphodiesterase activity) was identical. We also examined the capacity of these subsets to release various cytokines with or without polyriboinosinic and polyribocytidylic acid (Poly I:C) stimulation. There was no appreciable difference in their ability to release interferon (IFN), interleukin 1 (IL-1), and prostaglandin E (PGE) without stimulation, while IM produced slightly, but significantly, higher amounts of colony-stimulating factor (CSF) than RM. The amount of IFN released by IM in response to poly I:C was approximately three times higher than the amount of IFN released by RM. IL-1 was also released in higher amounts by IM than by RM in response to poly I:C. IM were also found to release more CSF than RM in response to poly I:C. In contrast, it was noted that IM secrete significantly less PGE response to poly I:C than do RM. These findings indicate that two purified human monocyte subsets, distinguishable by maturation markers, differ significantly in their ability to release various cytokines after stimulation; this difference may be relevant to potential in vivo roles of these immunoregulatory cells. PMID- 2984301 TI - Effects of nicotine on the functions of human polymorphonuclear leukocytes in vitro. AB - Effects of nicotine on migration, extracellular release of lysosomal enzymes, and superoxide anion (O-2) production of human polymorphonuclear leukocytes (PMN) were studied. Nicotine (5 X 10(-6) to 5 X 10(-4) M) had no effect on random migration, chemotaxis to fMet-Leu-Phe, nor on chemokinesis induced by fMet-Leu Phe. Nicotine, however, inhibited both extracellular release of lysosomal enzymes from PMN and O-2 production of PMN, both of which were induced by fMet-Leu-Phe and cytochalasin B. The inhibition of enzyme release and O-2 production by nicotine was not affected by atropine, hexamethonium, or acetyl beta methylcholine, suggesting a direct action of nicotine on PMN functions. It is presumed that nicotine does not affect PMN migration to inflammatory sites, but inhibits the microbicidal functions of PMN. Exposure to PMN to nicotine introduced into the body by smoking could suppress their functions. This might result in harmful influences on the host defense mechanism, including antitumor function. PMID- 2984300 TI - Loss of fibrinogen receptors from the platelet surface during simulated extracorporeal circulation. AB - In vitro recirculation of fresh human heparinized blood in an extracorporeal circuit with a membrane oxygenator decreased fibrinogen-induced platelet aggregation and diminished the number of fibrinogen receptors and glycoprotein IIb/IIIa (GPIIb/GPIIIa) antigenic sites on the platelet surface. In seven experiments, the mean +/- SD Km value for fibrinogen (i.e., molar concentration of fibrinogen required to cause 50% of the maximal rate of aggregation) was 1.58 X 10(-7) mol/L +/- 0.68 X 10(-7) mol/L. After recirculation, this value increased to 3.8 X 10(-7) mol/L +/- 1.94 X 10(-7) mol/L (P less than or equal to 0.025). The maximal aggregation rate of chymotrypsin-treated platelets decreased by 40% after 2 hours of recirculation (P less than or equal to 0.025). The number of fibrinogen receptors on platelets, which were treated with chymotrypsin after a recirculation, decreased from 41,370 +/- 24,000 to 13,230 +/- 10,230/platelet under the same conditions (P less than or equal to 0.025). The number of antigenic sites for monoclonal antibody reacting with GPIIb/GPIIIa complex of adenosine diphosphate-stimulated platelets decreased from 34,200 +/- 5,940 to 19,500 +/- 9,680/platelet after recirculation (P less than or equal to 0.025). Prostaglandin E1 (0.3 mumol/L) in the perfusion circuit preserved the ability of platelets to react with fibrinogen. In conclusion, the loss of fibrinogen receptors from the surface of platelet membranes results from the interaction of platelets with the surfaces of perfusion circuits. PMID- 2984303 TI - Regulation of macrophage lysosomal secretion by adenosine, adenosine phosphate esters, and related structural analogues of adenosine. AB - Zymosan particle-stimulated beta-galactosidase secretion by mouse peritoneal macrophages was found to be inhibited by micromolar concentrations of adenosine, AMP, ADP, and ATP. Inhibition by all four agents was increased to approximately 80% by adding erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA; 10 microM) an adenosine deaminase inhibitor, to the incubation medium. The inhibition of lysosomal enzyme secretion by ATP, ADP, and AMP was reversed by adding alpha, beta -methylene ADP (100 microM), a 5'-nucleotidase inhibitor, to the incubation medium. Inhibition by adenosine, however, was unaffected by alpha, beta methylene ADP indicating that the inhibition by AMP, ADP, and ATP only occurred after they had been converted to adenosine by cell surface phosphohydrolases, including 5'-nucleotidase. Theophylline, a competitive antagonist of the binding of adenosine to plasma membrane adenosine receptors, failed to reverse the inhibitory effect of adenosine indicating the probable site of adenosine action to be intracellular. Other purine nucleosides, e.g., guanosine, and several purine and ribosemodified structural analogues of adenosine also inhibited zymosan-stimulated beta-galactosidase secretion, while xanthosine and certain pyrimidine nucleosides, e.g., thymidine, were inactive in this respect. PMID- 2984304 TI - Cultured human alveolar macrophages from smokers with lung cancer: resolution of factors that stimulate fibroblast proliferation, production of collagenase, or prostaglandin E2. AB - Marked connective tissue remodelling involves both destruction and repair in inflammatory lung diseases. Throughout the remodelling event, it was reasoned that alveolar macrophages may release substances similar to those produced by blood monocyte-macrophages that affect fibroblast functions, ie, the interleukin 1 family of monokines (or cytokines). We have examined human alveolar macrophage cultures obtained after bronchoalveolar lavage of freshly excised lungs from heavy smokers with bronchial carcinoma. Crude culture media contained fibroblast proliferative activity and collagenase- and PGE2- production-stimulating activity. The main peak of these biological activities was located around approximately 18 kilodaltons (kD) on gel filtration chromatography. Resolution of this peak by high performance liquid chromatography showed the presence of three distinct peaks, with quantitative and qualitative differences in biological activities. This suggests the presence of heterogeneous factors. PMID- 2984305 TI - Retroviral induction of acute lymphoproliferative disease and profound immunosuppression in adult C57BL/6 mice. AB - We have shown that a mixture of murine leukemia viruses (MuLV) causes the acute onset of lymphoproliferation and immunosuppression when injected into adult C57BL/6 mice. The ecotropic/MCF (mink cell focus-inducing) mixture of MuLV stimulates polyclonal B lymphocyte proliferation and differentiation to antibody secreting cells. Serum Ig levels are elevated for all isotypes except IgA. The viral infection leads to a rapid decline in T lymphocyte responses to mitogens and alloantigens, as well as a decrease in helper cell activity. Specific antibody responses to both T-dependent and T-independent antigens are impaired, and the response of B lymphocytes to mitogens is abolished. The profound immunosuppression seems to be due to the MuLV-induced polyclonal activation of lymphocytes. No active suppression of normal lymphocyte responses by cells from virus-infected mice was observed. The disease induced by the LP-BM5 MuLV isolate thus seems a promising model for the study of lymphocyte activation and the mechanisms of retrovirus-induced immunosuppression. PMID- 2984306 TI - Human J chain gene. Structure and expression in B lymphoid cells. AB - As part of an ongoing investigation of the regulation of gene expression in B cell development, we have obtained a genomic DNA clone encoding the human J chain protein. The nucleotide sequence of exons encoding the mature protein defines a 137 amino acid primary sequence similar to that previously determined at the protein level. Probes from the gene have been used to analyze J chain expression in human cell lines corresponding to pre-B and B lymphocytes. J chain RNA was detected in two of six human pre-B cell lines and in 8 of 10 B cell lines expressing various Ig isotypes. The expression of the J chain gene is, thus, not tightly linked to IgM or IgA secretion. Our data do not, however, support the recent suggestion (7) that synthesis of J chain precedes that of mu chain in B lymphocyte differentiation. Because of the presence of nine candidate polyadenylation signals (AATAAA or AATTAAA) downstream of the C-terminal coding block of the J chain gene, the 3' end of the gene could not be determined from sequence data alone. To define the 3' end, J chain RNA from a human B lymphocyte line was used to protect an end-labelled DNA fragment from S1 nuclease digestion. The sequence 40 basepairs 5' of the functional polyadenylation site identified by these S1 experiments is homologous the same region of a previously reported mouse J chain complementary DNA clone. PMID- 2984308 TI - Altered distribution pattern of Na+-K+-ATPase and succinate dehydrogenase activities along the nephron in human acute post-transplant renal failure. AB - The catalytic activities of Na+-K+-ATPase and succinate dehydrogenase, marker enzymes for active salt reabsorptive capacity of renal basolateral plasma membranes and for respiratory capacity of mitochondrial cristae membranes, were studied in the maintenance phase of human acute post-transplant renal failure. Biopsies of 4 kidney-allografts taken at transplantation operation and additionally at different post-transplantation periods, either with good function or in various stages of dysfunction, were compared with the unaffected part of a human kidney nephrectomized due to hypernephroma. In single nephron segments, Na+ K+-ATPase activity was determined after microdissection by microfluorometry, and succinate dehydrogenase activity was determined by a microphotometric procedure in stained cryosections. In intraoperative and postoperative biopsies of a well functioning allograft, both Na+-K+-ATPase and succinate dehydrogenase activities did not differ from those of normal renal tissue. In contrast, the catalytic activities were found to be decreased in the distal tubules of 2 anuric allografts when compared with their intraoperative controls. In addition, succinate dehydrogenase activity was reduced in distal tubules of a recovering allograft. Catalytic activities appeared to be unaffected in glomeruli, proximal tubules, and collecting ducts. It is suggested that the predominant distal tubular alterations with regard to these parameters are a consequence of increased distal tubular vulnerability due to circulatory and metabolic conditions. PMID- 2984307 TI - Frequent biclonality and Ig gene alterations among B cell lymphomas that show multiple histologic forms. AB - Configurations of Ig gene DNA were examined in multiple biopsy specimens from seven cases of human B cell lymphoma that showed histologic differences among the specimens within each case. Analysis by Southern blot hybridizations with DNA probes for each of the three Ig loci revealed that the configurations of DNA within these loci were identical among the specimens in two of the cases. This result indicated the monoclonality of these lymphomas, despite differences in histology between biopsy specimens. In contrast, no common nongermline configurations of Ig gene DNA were detected among multiple biopsies in each of three other cases. Therefore, different histologies correlated with separate clones of proliferating B cells in these cases. In the last two cases, the configurations of light chain gene DNA were the same among biopsies in each case, consistent with a monoclonal origin in both lymphomas. However, differences were detected in the configuration of the heavy chain gene DNA. Analysis with a series of DNA probes of the mu heavy chain region indicated that the differences in the DNA configurations of the heavy chain genes from the biopsies probably arose from postrearrangement deletions of either the switch or constant regions of the mu gene. These studies indicate that, contrary to the conventional belief, individual tumors that contain different histologic types of lymphoma within the same patient frequently arise from separate clones of neoplastic cells. Furthermore, the heavy chain genes of monoclonal tumors may show postrearrangement deletions, often resulting from instability of DNA sequences within or around the mu switch region. PMID- 2984309 TI - Observations on ToxTrap silica gel breath capture tubes for alcohol analysis. AB - Experimental studies were carried out to investigate the accuracy, precision, and reliability of ToxTrap silica gel tubes relative to the capture, from Intoxilyzers, and subsequent analysis of alcohol derived from Simulator vapors or breath samples. Factors influencing analytical results, such as the presence of moisture in the tubes, were investigated. Comparisons were made between immediate, direct Intoxilyzer results and ToxTrap tube results obtained by a gas chromatographic technique. PMID- 2984310 TI - Free radical production from controlled low-energy fires: toxicity considerations. AB - Most fire departments respond within the first 5 min of notification of a fire. If fire victims are found at that stage by the firefighters, then incapacitation or death has occurred during the initial low-energy phase where smoke is being produced. Studies have shown that during this initial low-energy phase of the fire, gases commonly thought responsible for incapacitation or death are frequently not present in concentrations adequate to cause this result. In the current study free radicals, measured by electron spin resonance spectroscopy, were trapped in concentrations that we consider incapacitating, thus providing an explanation for "incapacitation without cause." This finding points the way to the design of more efficient temporary protective equipment for those who are in a high fire hazard environment, such as airline passengers, and suggests the idea of establishing a thermodynamic marker for the relative toxicity of building materials. PMID- 2984311 TI - Association of the encapsulation of Bacillus anthracis with a 60 megadalton plasmid. AB - Virulent typical strains (Shikan, Morioka, Shizuoka) and Pasteur vaccine strains (no. 1, no. 2-H, no. 2-17JB) of Bacillus anthracis harboured two plasmid species with molecular masses of 110 MDal and 60 MDal. All of the 110 MDal plasmids isolated from the various strains showed indistinguishable patterns of digestion with restriction endonucleases. All the 60 MDal plasmids were also indistinguishable. Strain Davis, which is encapsulated but is asporogenous and avirulent, harboured only the 60 MDal plasmid while three non-encapsulated vaccine strains (34F2, Smith, Mukteswer) harboured only the 110 MDal plasmid. Four non-encapsulated variant strains obtained from the encapsulated strains Shikan, Pasteur no. 1, Pasteur no. 2-17JB and Davis had lost the 60 MDal plasmid, suggesting that encapsulation of B. anthracis may be associated with the 60 MDal plasmid. PMID- 2984312 TI - Protein kinase activities associated with the virions of pseudorabies and herpes simplex virus. AB - Protein kinase has been extracted in soluble form from virions of pseudorabies virus using 10% NP40, 0.6 M-NaCl. Chromatographic analysis of the extract on DEAE cellulose and on phosphocellulose showed it to contain more than one kinase. The activity responsible for the phosphorylation of the major phosphoproteins (mol. wts. 120 000, 115 000 and 72 000) of virions was found to be similar in its properties to the host enzyme casein kinase II. Purified casein kinase II from ascites cells or from pig liver was able to phosphorylate heat-inactivated virions. In addition to the major phosphoproteins, active virion preparations were able to phosphorylate a minor low molecular weight phosphoprotein, incorporation into which could be stimulated by the addition of cyclic AMP to the assay. Purified host cyclic AMP-dependent protein kinase also phosphorylated this protein in heat-inactivated virions. Analysis of herpes simplex virus type 1 showed that the major phosphoproteins (VP12 and VP23) could be phosphorylated in heat-inactivated virions by added casein kinase II. One of these (VP12) together with a further minor phosphoprotein (VP14) could be phosphorylated by cyclic AMP dependent protein kinase. PMID- 2984313 TI - On the intracellular transport and the nuclear association of human cytomegalovirus structural proteins. AB - In cells productively infected with human cytomegalovirus (HCMV) AD169, large amounts of two viral proteins, the 150K major capsid and the 68K major matrix proteins, are continuously produced during the late phase of infection. In the present study, the mechanism for the intracellular transport of the 150K and 68K proteins was investigated. Infected cells were labelled for 30 min at 72 h post infection with [35S]methionine, chased for various periods of time at 37 degrees C, and fractionated into cytoplasmic and nuclear fractions. Immediately after 30 min of labelling, the 68K protein was already associated with the nuclear fraction. In contrast, the major proportion of the 150K protein remained in the cytoplasm for more than 1 h; the migration of the 150K protein was much slower than that of the 68K protein. Both the 150K and the 68K proteins were associated with the perinuclear cytoskeletal fraction in the process of migration. After migration into the nucleus, these proteins were resistant to extraction with DNase and high salt, indicating that they were associated with the nuclear skeleton (nuclear matrix). Effects of various inhibitors on the migration of the 150K protein showed that cycloheximide inhibited the transport of the 150K protein, but other inhibitors such as arabinosyl cytosine, cytochalasin D, colchicine or sodium azide did not. The results suggest that the cytoskeletal structure may play a role in the intracellular transport of HCMV structural proteins from the cytoplasm into the nucleus. PMID- 2984314 TI - Cloning and sequencing of the gene encoding the spike protein of the coronavirus IBV. AB - RNA sequences encoding the surface projection (spike) of the coronavirus infectious bronchitis virus, strain Beaudette, have been cloned into pBR322 using cDNA primed with a specific oligonucleotide. A 5.3 kilobase viral insert in the clone pMB179 has been identified. The region of this clone coding for the spike gene has been sequenced by the chain termination method, and we present here the first report of DNA sequence data for a coronavirus spike protein, the protein which forms the characteristic 'corona' after which the group is named. The amino acid sequence of the primary translation product, deduced from the DNA sequence, predicts a polypeptide of 1162 amino acids with a molecular weight of 127 006. This has many interesting features which confirm and extend our knowledge of this recently characterized membrane glycoprotein. The polypeptide is subsequently cleaved to S1 and S2, and partial amino acid analysis of the amino-terminus of the S1 polypeptide has been employed to locate the position of this terminus of S1 within the large open reading frame. The amino acid analysis also reveals the presence of an 18 amino acid putative signal sequence on the primary translation product which is not present on the mature S1 polypeptide. PMID- 2984315 TI - Encephalomyocarditis virus and diabetes mellitus: studies on virus mutants in susceptible and non-susceptible mice. AB - The so-called M-variant (especially subtype D) of encephalomyocarditis virus (EMCV) induces a diabetes-like syndrome in certain mouse strains which may serve as a model of insulin-dependent diabetes mellitus (IDDM) in man. The development and course of diabetes was influenced by a number of virus and host factors, among these being virus strain, virus dose, mouse strain, age, sex, and the host's immunological status. In a D-variant stock of EMCV, we found a virus plaque variant (PV 2) diabetogenic for DBA/2 mice, and at least one variant (PV 7) that did not affect carbohydrate metabolism. Although the diabetogenicity of PV 2 proved to be a genetically stable characteristic after further passages in vivo and in vitro, the incidence of diabetes varied somewhat (mean value 65% in 10-week-old DBA/2 mice infected with 10(5) p.f.u.). Both lower (10(1) or 10(3) p.f.u.) and higher (10(7) or 10(8) p.f.u.) virus doses led to a diminished incidence and severity of diabetes. In younger animals (5 weeks) transient hyperglycaemia often appeared, whereas in older animals (20 weeks) there was a higher rate of mortality. Histological examination of the islets of Langerhans in diabetes-susceptible (DBA/2) and resistant (C57BL/6) mice revealed that EMCV induced hyperglycaemia appeared to develop in parallel to islet cell damage. Even in diabetic animals, some unaffected islets were regularly found. This study demonstrates that EMCV mutants may have completely different biological effects and produce diabetes only in special circumstances. Host factors play a significant role in the development of diabetes. PMID- 2984316 TI - Herpes simplex virus ribonucleotide reductase induced in infected BHK-21/C13 cells: biochemical evidence for the existence of two non-identical subunits, H1 and H2. AB - In nearly all systems studied, ribonucleotide reductase consists of two non identical subunits. We present here the results of our study on herpes simplex virus (HSV) ribonucleotide reductase in favour of the existence of two subunits, H1 and H2, different from the mammalian subunits, M1 and M2. First, although the viral subunits could not be separated by Blue Sepharose chromatography (unlike mammalian subunits), they seemed to dissociate at very low protein concentration as suggested by the non-linear relationship between activity and low protein concentration. Second, pyridoxal phosphate (Pyr.P)-NaBH4 treatment and 4-methyl-5 amino-1-formylisoquinoline thiosemicarbazone (MAIQ) treatment of partially purified extract of mammalian ribonucleotide reductase which inactivated M1 and M2 respectively also inhibited the HSV ribonucleotide reductase. This activity could be restored by mixing Pyr.P-NaBH4-treated extracts with MAIQ-treated extracts of viral ribonucleotide reductase, suggesting that each treated extract contains one active subunit. Moreover, the addition of exogenous M1 or M2 subunits to one or the other of these two treated extracts did not produce any detectable reductase activity. Our interpretation of these results is that the two subunits H1 and H2 which could dissociate upon treatment did not form enzymically active hybrids with the mammalian subunits. Also, the higher degree of resistance to heat inactivation and to hydroxyurea of the viral reductase as compared to the mammalian enzyme suggests that H1 differs from M1 and H2 from M2. PMID- 2984317 TI - The cytotoxic response to murine cytomegalovirus. I. Parameters in vivo. AB - Combined in vivo and in vitro protocols for the generation of anti-murine cytomegalovirus (anti-MCMV) cytotoxic responses were investigated using BALB/c mice and syngeneic mouse embryo fibroblast target cells. Injections of doses of MCMV from 10(2.6) up to 10(6.1) p.f.u. into the hind footpad, harvest of draining popliteal lymph node cells after 6 to 8 days, followed by 4 days culture of these cells gave similar and optimal cytotoxic activity against MCMV-infected target cells. After injection of 10(4.6) p.f.u. into the hind footpad, MCMV was detectable at about 10(3) to 10(4) p.f.u. per popliteal node after 3 days, but became undetectable in most animals by 6 days. Cell numbers in the draining popliteal lymph node increased following MCMV inoculation into the hind footpad, but the extent of the increase was inversely related to virus dose and bore no relationship to the anti-MCMV cytotoxic potential of the cells. Lymph node cells applied directly to target cells upon harvest from MCMV-infected mice never gave detectable anti-MCMV cytotoxic activity at 2, 4, 6, 8, 10, 12, 17 or 19 days post infection, but lysis of uninfected syngeneic targets was obtained 4 to 8 days post-infection. PMID- 2984319 TI - Permeability changes elicited by influenza and Sendai viruses: separation of fusion and leakage by pH-jump experiments. AB - Permeability changes elicited in Lettre cells by influenza virus at pH 5.3 were maintained when the pH was shifted to 7.4. Permeability changes elicited by Sendai virus at pH 7.4 were maintained when the pH was shifted to 5.3. In each case permeability changes at the new pH were sensitive to inhibition by extracellular Ca2+. The time at which the pH was shifted was critical: if the shift was made prior to the onset of permeability changes, no subsequent changes occurred. It is concluded that the pH-sensitive event, namely virus-cell fusion, is related to the induction of permeability changes through attainment of some type of 'threshold' level of membrane damage. PMID- 2984318 TI - The cytotoxic response to murine cytomegalovirus. II. In vitro requirements for generation of cytotoxic T cells. AB - A cytotoxic response to murine cytomegalovirus (MCMV) was obtained by the culture of lymph node cells from mice inoculated with MCMV into both hind footpads 7 days previously. The cytotoxicity was mediated by Thy1.2+, Lyt2+, H-2-restricted effector cells and was virus-specific. Investigation of the in vitro conditions established that T cell proliferation was necessary for optimal generation of cytotoxicity, that proliferation was dependent upon Thy1.2+, Lyt2+ cell populations and that supernatants from concanavalin A-activated spleen cells enhanced the levels of cytotoxicity obtained. PMID- 2984320 TI - The experimental infection of chickens with mixtures of infectious bronchitis virus and Escherichia coli. AB - By inoculating chickens intranasally with a collection of strains of infectious bronchitis virus (IBV) of the Massachusetts serotype and of Escherichia coli of different serotypes, a pool of viral and bacterial strains was selected which, on inoculation, consistently produced a highly lethal disease closely resembling the natural disease produced by these two organisms. The conditions for reproducing the experimental disease were not rigorous in that, within broad limits, the size of the viral and bacterial inocula were not important; neither were the times at which both organisms were administered in relation to each other. The breed or strain of chicken used was important and the resistance of chickens to fatal infection increased with age. When the E. coli strains of the pool were inoculated intranasally without the IBV component, the chickens remained well; bacteriological examination of chickens inoculated with one of the E. coli strains, O18, revealed little evidence of invasion of the tissues or even of persistence of the inoculated E. coli strain in the upper respiratory tract. A minority of the IBV strains examined were lethal for chickens when inoculated without E. coli but many of them only produced a substantial mortality when the E. coli were included in the inoculum; IBV strains in this latter category included the vaccine strains H52 and H120. High concentrations of IBV strain M41 and E. coli O18 persisted in the upper respiratory tract for a number of days after they had been inoculated together. Much lower concentrations of IBV M41 were found in the internal organs, such as the spleen; E. coli O18 was only found in these sites in some of the inoculated chickens. Coliform organisms proliferated in the upper respiratory tract of chickens inoculated with IBV alone; they were rarely found in their internal organs. PMID- 2984321 TI - Enhancing antibodies, macrophages and virulence in mouse cytomegalovirus infection. AB - The infectivity of tissue culture-passed mouse cytomegalovirus (MCMV) for resident mouse peritoneal macrophages in the presence of serial dilutions of antiviral antibody was studied by fluorescent antibody staining and virus yields. Although MCMV was neutralized by high concentrations of antiserum, there was a twofold enhancement of infectivity by subneutralizing antibody concentrations. On further dilution of antiserum, significant neutralization appeared again. When F(ab')2 fragments of anti-MCMV IgG were used or when macrophages were pretreated with monoclonal antibody to Fc receptor, there was no enhancement, and no neutralization at high dilutions of antiserum. This suggests that both enhancement and high dilution neutralization are mediated via the Fc portion of IgG and Fc receptors of macrophages. Tissue culture-passed virus whose infectivity for macrophages was reduced by high dilutions of antibody was converted to a more infectious state by addition of anti-mouse immunoglobulin. Similar results were obtained with salivary gland virus, which is less infectious for macrophages and is coated with non-neutralizing antibody. Tissue culture passed virus is known to be less virulent for suckling mice and more infectious for macrophages than salivary gland-passed virus. When tissue culture-passed virus was coated with appropriately diluted antiviral antibody, not only was its infectivity for macrophages reduced, but it also became more virulent than control virus treated with normal mouse serum. These results are interpreted in terms of the optimal density of Fc on the virus-immunoglobulin complex in relation to the density of Fc receptors on macrophages. PMID- 2984322 TI - Association of virulence of murine cytomegalovirus with macrophage susceptibility and with virion-bound non-neutralizing antibody. AB - A comparison was made of the virulence in vivo and the infectivity in vitro for macrophages of (i) tissue culture-passed mouse cytomegalovirus (MCMV), (ii) salivary gland virus taken 3 weeks after infection (SGV 3w), and (iii) salivary gland virus taken 1 week after infection (SGV 1w). Salivary gland virus (3w) is known to be coated with non-neutralizing antibody, and is more virulent for newborn suckling mice and less infectious for macrophages than tissue culture passed virus (TCV). Properties of SGV 1w were similar to those of TCV. Infectivity of SGV 3w for macrophages was significantly enhanced by treatment with trypsin (10 micrograms/ml) and at the same time virulence was lost. When SGV 1w or TCV were treated with trypsin the infectivity for macrophages was unaltered as long as the inoculum was adjusted to contain the same number of p.f.u. as assayed in MEF. Trypsin-treated SGV 3w was neutralized not by rabbit anti-mouse IgG Fc, but by anti-Fab, whereas untreated virus was neutralized by both of these anti-mouse immunoglobulins. These results are discussed in terms of the association of virulence with virion-bound antibody and Fc receptors on macrophages. PMID- 2984323 TI - In situ localization of the major capsid protein during lytic infection by herpes simplex virus. AB - The intracellular localization of the major capsid protein (ICP5) of herpes simplex virus was studied during virogenesis. Except for a brief period at the onset of synthesis, this protein was found almost exclusively inside the nucleus. Its localization was not at random since 80% was tightly bound to the nuclear matrix as early as 4 h after infection. Discrete modifications of the fluorescence pattern occurred in an orderly fashion during the progression of the infection. Immunoelectron microscopic studies using Protein A-gold labelling demonstrated that this protein is synthesized on cytoskeleton-bound polyribosomes and accumulates in the central part of the nucleus where formation of viral capsids occurs; no gold particles were found in association with the peripheral chromatin or with the nucleolus. PMID- 2984325 TI - Comparison of Western Blot Analysis to microneutralization for the detection of type-specific herpes simplex virus antibodies. AB - A newly developed system (as described in the previous paper) employing separated herpes simplex virus (HSV) polypeptides electrophoretically transferred to nitrocellulose paper, Western Blot Analysis (WBA), was compared to microneutralization (MN) for the detection of type-specific antibody to HSV types 1 and 2 in 98 human sera. Sera containing HSV1-specific antibodies reacted at least twofold more densely (as quantitated by densitometric scanning) to HSV1 than HSV2 polypeptides, while sera with HSV2-specific antibodies reacted twofold more densely to HSV2 polypeptides. The reliability of this system was determined (1) by analysis of sera from patients with true primary herpes simplex infections, (2) by adsorption studies that removed cross-reacting antibodies, and (3) by comparison with neutralization. Of 36 sera found to have HSV1-type specific antibody by WBA, 83% had MN ratios of less than 85 (indicating HSV1 antibody), while of 28 sera found to have HSV2-type-specific antibody by WBA, 89% had MN ratios of greater than 85 (indicating HSV2 antibodies). The MN ratio obtained on sera felt to have both HSV1-type-specific and HSV2-type-specific antibody by WBA ranged from 67 to 160. WBA agreed more closely with clinical isolates and adsorption studies than MN. This system not only accurately determines the HSV-type-specific antibody status of patients but provides important information on the specific immunogenic proteins. PMID- 2984324 TI - Identification and typing of herpes simplex virus types 1 and 2 by monoclonal antibodies, sensitivity to the drug (E)-5-(2-bromovinyl)-2'-deoxyuridine, and restriction endonuclease analysis of viral DNA. AB - With development of antiviral drugs, the need to identify a virus as to drug sensitivity becomes increasingly of importance. The compound (E)-5-(2-bromovinyl) 2'-deoxyuridine (BVDU) has been shown to be much more inhibitory to the replication of herpes simplex virus type 1 (HSV-1) and varicella-zoster virus as opposed to herpes simplex virus type 2 (HSV-2). We have typed over 170 isolates, using an immunofluorescent technique and sensitivity to the drug BVDU. These results were then compared to the typing of isolates by analysis of viral DNA after restriction endonuclease digestion (EcoRI). Without exception the results were in agreement between the monoclonal antibody results and sensitivity to the drug BVDU. Furthermore, the typing with monoclonal antibodies was also in excellent agreement with the DNA analysis. Only those isolates inhibited with BVDU showed DNA characteristics of HSV-1 and reacted only with the S-200 antibody. On the other hand, those isolates which reacted with the monoclonal antibody S-141 were insensitive to BVDU, and again this was in agreement with the DNA analysis. These results could provide the basis for developing a diagnostic test using the two monoclonal antibodies to type either isolates or direct smears and to use the results as a basis for possible drug therapy. PMID- 2984326 TI - Genetic heterogeneity of the human papovaviruses BK and JC. AB - We have examined the structure and infectivity of BKV and JCV genomes from prototype strains after cell culture passage and of BKV genomes from primary isolates. Genomic structures were determined by restriction endonuclease analysis of molecularly cloned DNA. Infectivity was determined by transfection of the cloned genomes into urine-derived epithelial cells and assaying for viral proteins and virus production. Prototype BKV DNA, which was cloned after 14 passages in three different cell lines, contained no alterations in restriction enzyme sites and was infectious. In contrast, prototype JCV acquired changes in the late region of the genome during passage in cell culture and the cloned DNA was not infectious. Urine-derived cells were used to isolate virus from the urine of two renal transplant patients and one asymptomatic individual. The genome of the virus isolated from the normal individual was indistinguishable from prototype BKV except for a 60-base pair deletion, which was localized between 0.62 and 0.72 map units. Two isolates from transplant patients differed from each other and from prototype BKV at a number of restriction enzyme cleavage sites located in the early region and were infectious. Genomes containing deletions from 100 to 600 base pairs were also cloned but were not infectious. PMID- 2984327 TI - Transfusion-acquired cytomegalovirus infection in cardiac surgery patients. AB - The incidence of transfusion-acquired primary cytomegalovirus (CMV) infection was studied in 483 cardiac surgery patients. Ninety-six patients (20%) were found to lack antibody to CMV [CMV Ab(-)] as measured by radioimmunoassay. Sixty-eight CMV Ab(-) were followed by viral culture and/or serology from eight weeks to one year after transfusion. Transfusion requirements in CMV Ab(-) patients were as follows: whole blood/packed red blood cells, mean 4.7 +/- 2.6 units; platelets (20 patients), 6.9 +/- 3.8 units; fresh frozen plasma (25 patients), mean 3.3 +/- 1.6 units. Forty-nine percent of 235 donor units tested had antibody to CMV. One donor unit (0.4%) had CMV-specific IgM. This was not associated with CMV infection in the recipient. One patient (1.5%) demonstrated evidence of seroconversion to CMV during the follow-up period. This is significantly less than reported in previously published studies (P less than .01). Serological methods used, the age of the transfused blood, the immune status of the transfusion recipient, and the administration of passive antibody in fresh frozen plasma are factors that may be responsible for the low incidence observed. PMID- 2984328 TI - Secretion of polyalbumin receptors in vitro. AB - The nature of hepatitis B surface antigen (HBsAg)-associated receptors for polymerized human serum albumin (pHSA-R) and their relationship to hepatitis B e antigen (HBeAg) and human serum proteins have not been defined. We studied by radioimmunoassay and by electron microscopy HBsAg-associated pHSA-R secreted in vitro by a human hepatocellular carcinoma cell line (PLC/PRF/5) and by mouse 3T3 fibroblasts after transfection with cloned hepatitis B virus (HBV) DNA (4.10 cells). PLC/PRF/5 cells expressed only HBsAg, whereas 4.10 cells secreted also HBeAg. There was no significant difference in the production of HBsAg, HBeAg, and pHSA-R when the cells were cultured in the presence or absence of fetal calf serum. Secretion of pHSA-R by the two cell lines for a given amount of HBsAg was equal irrespective of the presence or absence of HBeAg. Supernatants from both cell lines grown in serum-free medium did not contain any Clq or albumin when tested by immunodiffusion. The ability of a transfected mouse cell line to produce HBsAg with pHSA-R activity strongly suggests that pHSA-R is coded by the HBV genome and does not depend on the presence of human serum proteins. In addition, our findings fail to demonstrate any correlation between HBeAg production and pHSA-R. PMID- 2984329 TI - Polyalbumin receptors, hepatitis B surface antigen (HBsAg), and HBsAg/IgM complexes in HBsAg positive patients with and without delta superinfection. AB - Receptors for polymerized human albumin are found at high titres during high level hepatitis B virus (HBV) replication and in small amounts in chronic low level infection. Complexes between hepatitis B surface antigen (HBsAg) and IgM without specificity for HbsAg are expressed in a pattern similar to that of receptors. Anti-albumin antibodies could be involved in their formation. Delta infection depresses the synthesis of gene products of HBV. To assess whether delta modifies the expression of receptors on HBsAg and the level of HBsAg/IgM complexes, and if anti-albumin antibodies are actually part of the complex, we tested sera from 86 subjects with acute and chronic HBV infection. Our findings show that the amounts of circulating receptors and HBsAg/IgM are proportional to the concentration of HBsAg and thus probably to the degree of viral replication. We did not find any correlation between circulating anti-albumin antibodies of the IgM class and HBsAg/IgM complexes. Delta infection depresses HBsAg synthesis and causes a related decrease in receptors and HBsAg/IgM titres if superimposed on the more active stage of HBV infection. When HBsAg, receptors, and HBsAg/IgM are at low levels, no further depression is caused by delta infection. HBsAg/IgM titre is not enhanced by presence of anti-delta antibodies, thus excluding a role of the latter in forming these complexes. PMID- 2984330 TI - In vivo and in vitro studies on the potentiation of muscarinic receptor stimulation by alaproclate, a selective 5-HT uptake blocker. AB - Alaproclate (10-60 mg/kg) injected i.p. into male mice potentiated and prolonged the oxotremorine and physostigmine-induced tremor in a dose-dependent manner. Atropine completely blocked the tremor caused by oxotremorine or physostigmine both in the presence and absence of alaproclate. Pretreatment with the 5-HT receptor antagonist metitepine completely blocked the enhancement of oxotremorine induced tremor caused by alaproclate. Biochemical studies indicated that the above effects cannot be explained by assuming that alaproclate a) acts as a cholinergic agonist, b) inhibits the acetylcholine esterase, c) interferes with choline uptake or acetylcholine synthesis, or d) directly potentiates the release of acetylcholine. In ligand binding studies alaproclate was found to be a weak competitive inhibitor of muscarinic antagonist binding to membranes from the rat cerebral cortex, rat striatum, human cerebral cortex and human striatum. (Ki approximately 28-40 microM in all four tissues). The present results suggest that alaproclate may potentiate muscarinic responses by a mechanism involving serotonergic receptor mechanisms rather than by a direct interaction with the muscarinic cholinergic receptors. PMID- 2984331 TI - Fenfluramine stimulation of serum cortisol in patients with major affective disorders and healthy controls: further evidence for a central serotonergic action of lithium in man. AB - In order to investigate the influence of lithium long-term medication on serotonergic neurotransmission, fenfluramine stimulation (FFS) was used for the assessment of hormonal effects under serotonergic control. The cortisol plasma concentration following FFS was examined between 8 a.m. and 1 p.m. in 11 manic depressive subjects under lithium prophylaxis and in 8 untreated euthymic patients. In addition, 11 healthy subjects with FFS, and 12 other subjects without FFS were investigated. The basal cortisol concentrations show considerable variation. Those of the lithium patients were in general found lower than those of the control groups. In both, the controls and the manic-depressive patients without lithium medication, no gross deviation from the expected physiological decline of morning cortisol values was found. A subtle effect of FFS in healthy subjects could be observed. In the lithium patients, however, a significant inversion of the cortisol secretion pattern with a steep increase between 10 and 12 a.m. could be demonstrated. It is concluded that FFS and lithium long-term medication exert an agonistic influence onto central serotonergic neurotransmission. Pharmacological challenge with fenfluramine may prove to be a useful tool for the investigation of serotonergic mechanisms in biological psychiatry. PMID- 2984332 TI - Intravenous methylprednisolone for multiple sclerosis in relapse. AB - A randomised comparison is made between methylprednisolone, 1 g intravenously daily for 7 days, and a standard ACTH regime for the treatment of multiple sclerosis in acute relapse. It is found that methylprednisolone produces a more rapid clinical improvement than ACTH but confers no longer term benefit when the two treatments are compared at 3 months. It is proposed that intravenous methylprednisolone does have a role to play in the management of a patient with an acute relapse of multiple sclerosis. PMID- 2984333 TI - Hemifacial spasm associated with a mixed benign parotid tumour. PMID- 2984334 TI - Familial clustering of the ataxic form of Creutzfeldt-Jakob disease with Hirano bodies. AB - A family cluster of the ataxic form of Creutzfeldt-Jakob disease with one probable and two autopsy proven cases that occurred in a single generation between 1974 and 1982 is reported. The clinical characteristics of the cases are closely similar to those of kuru patients, with a fair correlation between the prominent truncal ataxia and the intense devastation of the cerebellar cortex most marked in the vermis. Pathologically, the marked hippocampal involvement rarely seen in typical transmissible Creutzfeldt-Jakob disease and the finding of Hirano bodies in the Ammon's horn without specific Alzheimer's senile changes are noteworthy features. PMID- 2984335 TI - Inclusion body myositis: a case with associated collagen vascular disease responding to treatment. AB - Patients with inclusion body myositis demonstrate characteristic histological and electronmicroscopical abnormalities in muscle and are generally considered refractory to treatment. A patient with inclusion body myositis is described with evidence of associated autoimmune disease, who responded to steroids. PMID- 2984336 TI - A note on the mechanism of resistance to anoxia and ischaemia in pathophysiological mammalian myelinated nerve. AB - Computer simulation of the action potential in myelinated nerve fibres show that the metabolic cost of conduction of an impulse is less than normal in a slightly depolarised fibre. This would account, at least in part, for the greater resistance to ischaemia and anoxia of nerves from diabetics and other pathophysiological conditions. PMID- 2984337 TI - The distribution of 2':3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in the CNS of normal (+/+) and shiverer (Shi/Shi) mice. AB - Immunohistochemical techniques were utilized to localize the putative myelin enzyme 2':3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in the central nervous system (CNS) of normal (+/+) and Shiverer (Shi/Shi) mice (Mus musculus). CNP appeared to be only associated with myelinated nerve fibers in the CNS (corpus callosum, subcortical white matter, caudate nucleus, cerebellum and medulla oblongata) of +/+ mice. However, little or no immunostaining was observed in the same regions of the CNS of Shi/Shi mice, although these mice have normal levels of a CNS CNP activity. Unexpectedly, oligodendrocytes (cell periphery) were not stained for CNP in either +/+ or Shi/Shi mice, although erythrocytes were immunostained. PMID- 2984338 TI - Fabry's disease with partially deficient hydrolysis of ceramide trihexoside. AB - A report is presented on biochemical studies of the fibroblasts from a 26-year old man with Fabry's disease whose clinical picture was atypical. The patient had severe pain in the extremities, but no angiokeratomas, corneal clouding or hypohidrosis. The trihexosylceramidase activity in the fibroblasts in vitro was deficient. The level and Km value of the residual activity were similar to levels seen in typical Fabry's patients. However, fibroblasts from the patient cultured in medium supplemented with [3H]ceramide trihexoside hydrolyzed the labeled ceramide trihexoside much higher than did cells from typical Fabry's patients, implying that the patient has a partial defect in hydrolysis of trihexosylceramide in cultured fibroblasts. PMID- 2984339 TI - In situ activation of murine peritoneal macrophages by cyclohexyl-1,3-dioxepin and 4-methyl-2-pentenoyl maleic anhydride copolymers. AB - Peritoneal macrophages (PM theta) from mice treated intraperitoneally with the unique polyanionic compounds cyclohexyl-1,3-dioxepin maleic anhydride copolymer (CDA-MA) and 4-methyl-2-pentenoyl maleic anhydride copolymer (MP-MA) had tumoricidal activity against Lewis lung tumor cells. 5'-Nucleotidase ectoenzyme activity, which had previously been associated only with nontumoricidal, resident PM theta, was elevated in tumoricidal PM theta elicited with CDA-MA and MP-MA. Cell counts and differentials performed on peripheral blood leukocytes and PM theta populations from CDA-MA- and MP-MA--treated mice more closely corresponded to those of normal mice than mice treated with the conventional PM theta activating agents pyran and Corynebacterium parvum. In addition, the lysosomal peroxidase activity in PM theta after administration of CDA-MA and MP-MA remained at levels comparable with normal resident PM theta, while an influx of peroxidase positive macrophages was observed after administration of pyran and C. parvum. Inoculation of CDA-MA and MP-MA into mice bearing Lewis lung carcinoma showed a significant increase in median survival time compared with control mice, as well as an increase in the percentage of mice that survived greater than 90 days. Taken together, these data suggest that CDA-MA and MP-MA activated PM theta in situ and prolonged survival against primary transplanted Lewis lung carcinoma. PMID- 2984340 TI - Human leukocyte interferon as part of a combined treatment for previously untreated small cell lung cancer. AB - Human leukocyte interferon, HuIFN-alpha (LE), has been tested in combination with radiotherapy and chemotherapy for previously untreated small cell lung cancer. Nine patients with limited disease received high-dose IFN followed by a low-dose regimen; and six patients had a low-dose regimen from the beginning. The high dosage of IFN consisted of 800 X 10(6) IU given as a continuous intravenous infusion for 5 days, followed by 6 X 10(6) IU i.m. three times weekly. If the first site of disease progression was local or in a central nervous system location, radiotherapy (55 Gy/20 F/7 weeks locally and/or 30 Gy/10 F/2 weeks whole brain) was applied and IFN was continued. Chemotherapy was administered only if there was disease dissemination outside the chest. Three patients achieved minor response for as long as 20, 25, and 42 weeks, respectively, with IFN alone. Three of five complete responders to IFN-radiotherapy died 18, 33, and 41 weeks from the start of IFN treatment without chemotherapy. Autopsy did not reveal macroscopic or microscopic tumor at any site, but there was severe radiation pneumonitis. Four of nine patients were administered chemotherapy subsequent to IFN-radiotherapy because of disease dissemination. The median length of survival of the entire group was 41 weeks. On the low-dose regimen, one patient achieved partial response with IFN alone (duration, 12 weeks); of five evaluable patients three achieved complete remission and two partial remission to IFN-radiotherapy, and one of the three complete responders to IFN-radiotherapy died of severe radiation pneumonitis at 21 weeks from the start of IFN treatment. No tumor was detected at autopsy. The study is in progress. Average survival at present is 33 weeks. The results derived from both our studies suggest a growth delaying effect of HuIFN-alpha (Le) on small cell lung cancer. They also suggest potentiation of radiation by HuIFN-alpha (Le). Memory and psychomotor dysfunction, fatigue, and anorexia were dose limiting with both short-duration, high-dose and long-duration, low-dose IFN therapy. We feel that IFN, as part of a combined multimodality treatment of small cell lung cancer, may play a role by delaying metastatic dissemination. PMID- 2984341 TI - T cell growth factor required for optimal induction of T cell growth factor receptor expression in phytohemagglutinin-stimulated T cells. AB - Proliferation of normal human T cells in vitro requires activation of resting T cells by lectin or antigen. This stimulation initiates a series of events which includes elaboration of T cell growth factor (TCGF), expression of TCGF receptors, and, ultimately, cellular proliferation. We sought to determine if TCGF was required for expression of the TCGF receptor in phytohemagglutinin (PHA) stimulated normal human T cells. Utilizing dexamethasone (DEX), a known inhibitor of TCGF production, reductions in T cell proliferation, TCGF production, and TCGF receptor expression, as measured by TCGF adsorption and Tac acquisition, were demonstrated after PHA stimulation. When exogenous partially purified TCGF was added to DEX-containing cultures, the DEX inhibition of proliferation and TCGF receptor expression was completely reversed. These experiments were reproduced utilizing both highly purified TCGF from the Jurkat cell line and purified TCGF synthesized by bacteria from cloned TCGF DNA. Short-term experiments showed TCGF to be capable of restoring Tac antigen expression after DEX inhibition in the absence of cellular proliferation. These results indicate that TCGF is required for optimal expression of Tac antigen-associated TCGF receptors in PHA-activated T cells. PMID- 2984342 TI - Detection of bone marrow metastasis in small-cell lung cancer by monoclonal antibody. AB - A murine monoclonal antibody against a surface antigen of small-cell carcinoma of the lung (SM1 antibody) was investigated for its use in detecting bone marrow metastasis. Bone marrow cells of healthy volunteers and of patients with small cell carcinoma of the lung (SCCL) were examined for reactivity with SM1 antibody and indirect immunofluorescence and the results compared to conventional histochemical staining (Wright-Giemsa stain of bone marrow aspirates and hematoxylin-eosin stains of bone marrow biopsies). No SM1 reactivity was found in marrow cells of eight healthy volunteers. Thirty-six samples from 33 patients with SCCL were examined; tumor involvement was found in 69% by SM1 antibody and in 16% by histochemical stains. All bone marrow samples from patients with SCCL that were unreactive with SM1 antibody also showed no evidence of tumor involvement by histochemical stains. Samples of 29 patients were investigated at initial staging; SM1 reactive cells were found in 50% of 16 patients with limited disease and in 77% of 13 patients with extensive disease. Overall, the proportion of patients recognized to have disseminated disease at diagnosis was increased from 45% to 72% by monoclonal antibody staining. Indirect immunofluorescence with SM1 antibody allows detection of bone marrow metastasis of SCCL that cannot be seen by conventional morphology and can identify disseminated disease in patients otherwise staged limited disease. PMID- 2984343 TI - Risk-benefit relationships in cancer clinical trials: the ECOG experience in non small-cell lung cancer. AB - Although there is widespread recognition of the need to critically evaluate risks and benefits for patients participating in clinical trials, the actual implementation can be a difficult task. As an illustration of the analytic difficulties, we reviewed the experience of the Eastern Cooperative Oncology Group (ECOG) in advanced (inoperable) non-small-cell lung cancer over the past ten years (1973 to 1983). Of 2,714 ECOG patients analyzed, 15% showed objective tumor response. Median survival of all patients was 4.2 months, with approximately one half of patient's survival spent on protocol treatment. Thirty nine percent of patients experienced at least one episode of severe or worse toxicity from therapy. Chemotherapy impact on this disease was assessed by examining trends in patient outcomes over the decade studied and by comparisons with patients receiving no treatment from earlier Veterans Administration Lung Protocols. Introducing more intensive chemotherapy regimens over this period appears to have resulted in some improvement in survival and response to therapy, but at the expense of greater toxicity. Despite modest survival gains achieved by these evolving trials, the community benefit from such trials seems clear, both in identifying ineffective therapies (and avoiding their general use) and as an important step in developing effective regimens. However, the decision for an individual patient to participate in a trial may involve difficult trade-offs between risk and benefit. This study suggests the need to identify subgroups of patients unlikely to benefit from trial participation and stresses the importance of incorporating patient preferences in the final decision. Some of the problems in patient communication are discussed. PMID- 2984344 TI - Anaplastic Wilms' tumor: clinical and pathologic studies. AB - A review of almost 1,200 children participating in the first and second National Wilms' Tumor Study (NWTS-1 and -2) has demonstrated a number of significant differences in the clinical presentation and response to therapy of anaplastic and nonanaplastic Wilms' tumor. Compared to their counterparts, children with anaplastic Wilms' tumor were generally one to two years older at diagnosis, more were non-white, and more had lymph node metastases at diagnosis. Consistent with previous studies, children with anaplastic Wilms' tumor survived for a significantly shorter time than those with non-anaplastic Wilms' tumor. A hopeful outlook, however, was suggested by the NWTS-2 experience since the more aggressive chemotherapies used in this study appear to have substantially improved the survival of patients with diffusely anaplastic tumors. Also, the survival of NWTS-2 patients with anaplastic Wilms' tumor was determined in part by clinicopathologic stage. It may be possible therefore to refine therapy according to stage so as to provide children with localized disease a chance for cure with fewer untoward treatment-related sequelae. PMID- 2984345 TI - Prognosis for Wilms' tumor patients with nonmetastatic disease at diagnosis- results of the second National Wilms' Tumor Study. AB - Multivariate statistical methods were used to study prognosis for 632 patients entered on the second National Wilms' Tumor Study who had nonmetastatic, unilateral disease at diagnosis. Separate analyses were conducted for each of four endpoints: abdominal recurrence, distant metastasis, relapse without regard to site, and death. The two most important predictors for metastasis and general relapse were an unfavorable (anaplastic or sarcomatous) histology and the presence of microscopically confirmed disease in the regional lymph nodes. Operative spillage of tumor increased the rates of abdominal recurrence and death, even after accounting for histology and lymph node effects. The presence of a tumor thrombus in the renal vein or IVC increased the risk of metastasis, and intrarenal vascular invasion was associated with general relapse after accounting for histology, lymph nodes, and spillage. However, these latter associations were weaker, and some uncertainty remains regarding the true prognostic import of such findings due to a high degree of collinearity among variables. By contrast to the results of a similar data analysis for the first National Wilms' Tumor Study, specimen weight had no bearing on outcome, and the effects of age at diagnosis were entirely explained by the association of age with other more critical factors. PMID- 2984346 TI - Second malignant neoplasms in children: an update from the Late Effects Study Group. AB - This paper presents an update from the Late Effects Study Group on 292 cases of second malignant neoplasms (SMN) occurring in individuals who were diagnosed with their first neoplasm in childhood. Data are presented regarding the types of first and second neoplasm, the therapy administered, and the predisposing factors. Of the 292 cases (308 SMN), the most common primary was retinoblastoma followed by Hodgkin's disease, soft-tissue sarcomas, and Wilms' tumor. This is not similar to the relative frequency of these cancers in children but rather reflects specific risk factors. Bone sarcomas were the most common SMN among the 208 SMN developing in previously irradiated sites while acute leukemia was the most common SMN unassociated with radiation. Known predisposing conditions to cancer were present in 73 cases; retinoblastoma was the most common of these, followed by neurofibromatosis. There were ten patients with three and three patients with four malignant neoplasms. In 14 patients, the cause of SMN was not suggested by known risk factors as these patients had negative family histories and received no radiation or chemotherapy. We note, therefore, that although most cases of SMN in survivors of childhood cancer can be attributed to radiation, genetic disease, chemotherapy, or combinations of these, unrecognized predisposition or chance may also play a role. PMID- 2984347 TI - Signal transmission in the catfish retina. I. Transmission in the outer retina. AB - Extrinsic current, either pulsatile or white-noise modulated, was injected into the (cone) horizontal-cell soma and axon, and resulting responses were recorded from nearby points. In the case of white-noise inputs, signal transmission between the two points was characterized by Wiener kernels. The signal transmission within the lamina, the S-space, formed by the (cone) horizontal-cell somas and axons is quasi-linear and very fast, indicating that the laminae are purely resistive networks within the frequency range of the light-evoked response. There exists signal transaction between the lamina formed by the somas and axons. The forward transmission is constant gain, low pass, but there is a filter for the reverse transmission to impede the backflow of high-frequency components. Signals in the horizontal-cell soma are transmitted to the bipolar cells. The transmission is sign noninverting for the on-center bipolar cells and sign inverting for the offcenter cells. The transmission is quasi-linear excluding complex mechanisms in the transmission. We believe that the forward and direct transmission of signals from the horizontal to bipolar cells is the most straightforward interpretation of the observation. The transfer functions between the horizontal and bipolar cells differ considerably from one bipolar cell to the next. PMID- 2984348 TI - Signal transmission in the catfish retina. II. Transmission to type-N cell. AB - Responses from channel catfish type-N (sustained amacrine) cells were evoked either by step changes in illuminance, i.e. brightening or dimming from a mean illuminance, or by a white-noise modulated light stimulus. Current injected into the horizontal-cell soma or axon produced responses in type-N cells that were very similar to those produced by light stimuli. Light- and current-evoked responses had linear and second- and third-order nonlinear components; the former contributed 40-50%, whereas the latter contributed 20-30% to the total response. The remainder of the response could have been due to higher-order nonlinearities or to intrinsic as well as extrinsic noise. Nonlinear components in the light- and current-evoked responses were sharp transient peaks, which were prominent in white-noise-evoked responses, and oscillatory wavelets. The high-frequency components in the cell's response, which result from nonlinearity, were absent in the responses from bipolar and horizontal cells. The nonlinear responses were predicted by the second- and third-order kernels. The type-N cell response was complex because the response had both linear and nonlinear components, and because of the complexities of second- and, probably, third-order kernels. The cell's complex response reflects the complex nature of the cell's function as well as its synaptic organization. PMID- 2984349 TI - Signal transmission in the catfish retina. III. Transmission to type-C cell. AB - Current injected into horizontal-cell somas and axons produced transient (on-off) depolarizations from type-C cells (commonly known as transient amacrine cells) similar to those produced by light. Both the light- and current-induced responses had very small linear components and nonlinear components as represented by the second-order kernels, which reproduced the cell's response with a reasonable degree of accuracy. The second-order kernels were well defined and stereotyped. The quadratic nature of the nonlinear component is reflected in the frequency doubling response as well as the very steep input-output relationship of the cell. Type-C cell's responses evoked by light and current differed in a subtle but distinct fashion, and this difference appeared in the signature of the second order kernels. The light-produced kernels had two diagonal positive peaks and off diagonal valleys ("four-eye" structure), whereas the current-produced kernels had a single on-diagonal positive peak with off-diagonal negative valleys ("three eye" structure). The difference in the kernel configuration was reflected in the cell's step-evoked response. Some type-C cells produced faster responses whereas others produced slower responses, whether evoked by light or by current. Our past and present results show that type-C cells produce a very nonlinear response that is not necessarily complex. PMID- 2984350 TI - Mechanisms of pattern generation underlying swimming in Tritonia. IV. Gating of central pattern generator. AB - Swimming behavior in the marine mollusc Tritonia diomedea is episodic, consisting of a series of alternating dorsal and ventral flexions initiated by a brief sensory stimulus. The swim motor pattern is generated by a network formed of four groups of premotor interneurons: cerebral cell 2 (C2), dorsal swim interneurons (DSIs), and two types of ventral swim interneurons (VSI-A and VSI-B). The initiation and maintenance of swimming depends on the establishment of a long lasting ramp depolarization in both the premotor, pattern-generating interneurons, and the motor neurons (i.e., flexion neurons). Voltage clamp was used to measure the membrane current responsible for the ramp depolarization. In all cell classes the current had two components: a tonic inward current, which decayed as the swim progressed, and phasic inward current waves, which provided the synaptic drive during each swim burst. The ramp current in the flexion neurons and in C2 was generated largely by activity within the interneuronal pattern-generating network (PGN). The ramp current could be mimicked by driving activity in the pattern-generating interneurons. In VSI-B, the tonic component of the ramp current was independent of activity within the PGN and appeared to be derived from the long-lasting effect of an extrinsic input. The phasic components of the ramp, however, were dependent on PGN activity. The phasic inward current waves were blocked when pattern generation was prevented. In addition, phasic inward currents similar to those occurring during swimming could be produced by driving the C2. The tonic component of the ramp current in a DSI was dependent both on extrinsic inputs and PGN activity. Extrinsic inputs appeared to control the first 10-15 s of the tonic current. At longer times, activity within the DSI population itself maintained the ramp current. When one DSI was driven in a quiescent preparation, all other DSIs were inhibited, yet the DSIs are known to be coupled by monosynaptic, reciprocal excitatory synapses. This effect could be explained by the action of an unidentified inhibitory interneuron (I-neuron), which was excited by DSIs and in turn inhibited all other DSIs. The DSIs were therefore coupled reciprocally by both monosynaptic excitation and polysynaptic inhibition. Activity in C2 switched the DSI-DSI interaction from inhibition to excitation by inhibiting the I-neuron.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984351 TI - Distribution of oligosynaptic group I input to the cat medial gastrocnemius motoneuron pool. AB - To characterize the oligosynaptic group I afferent input to the cat medial gastrocneumius (MG) motoneuron pool, the medial branch of the tibial nerve (MTIB: flexor digitorum and hallucis longus, popliteus, tibialis posterior and interosseous nerves), the nerves to flexor digitorum and hallucis longus (FDHL), or the nerves to the quadriceps muscles (QUAD) were stimulated at submaximal group I strength while recording intracellularly from MG motoneurons. Since previous work indicates that stimulation of these nerves at group I strength produces no significant monosynaptic Ia excitation or Renshaw inhibition of MG motoneurons, group I effects were assumed to be predominantly, though not exclusively, due to the action of Ib-fibers. Evidence supporting this assumption is presented in the following paper. MTIB, FDHL, and QUAD postsynaptic potentials (PSPs) were most commonly inhibitory. Since the MTIB, FDHL, and QUAD nerves are composed predominantly of fibers innervating muscles with extensor action, their inhibitory effect on MG motoneurons is consistent with previous findings that stimulation of Ib-afferents in nerves to extensor muscles produces di- and trisynaptic inhibition of extensor motoneurons. However, excitatory effects were observed in about one third of the motoneurons, indicating that oligosynaptic group I input is not homogeneously distributed within the MG motoneuron pool. Variations in QUAD, FDHL, and MTIB PSP pattern and amplitude were correlated with variations in the PSP pattern evoked by stimulation of the sural nerve: excitatory oligosynaptic group I PSPs generally appeared in motoneurons receiving excitatory cutaneous (sural nerve) input, whereas inhibitory PSPs generally appeared in motoneurons receiving some inhibitory cutaneous input and were largest in motoneurons receiving predominantly inhibition from the sural nerve. These variations in QUAD, FDHL, and MTIB PSP pattern and amplitude were not due to variations in resting potential and were only partly due to variations in intrinsic motoneuron properties or motoneuron "type." Our results indicate that activation of these cutaneous and group I muscle afferents can exert similar effects on the MG motoneuron pool. Moreover, the presence of a strong correlation between the distributions of cutaneous and oligosynaptic group I PSPs within a single motoneuron pool is consistent with the results of previous studies that have shown that some of the input to motoneurons from these peripheral afferents is mediated through common interneurons. PMID- 2984352 TI - Use-dependent depression of IPSPs in rat hippocampal pyramidal cells in vitro. AB - We have used intracellular recording techniques to study the use-dependence of evoked inhibitory postsynaptic potentials (IPSPs) in rat CA1 hippocampal pyramidal cells. We determined reversal potentials and conductance changes associated with IPSPs and responses to directly applied gamma-aminobutyric acid (GABA). The IPSP depression could be seen after a single conditioning stimulus. This depression appeared to be due primarily to a 50% decrease in IPSP conductance (gIPSP). Trains of stimulating pulses (50 pulses at 5 or 10 Hz) produced more pronounced effects than a single conditioning pulse. Suprathreshold repetitive stimulation of stratum radiatum (SR) produced epileptiform burst firing and greater depression of IPSPs than did alvear (ALV) or subthreshold SR stimulation. During suprathreshold SR stimulation the IPSP was nearly abolished and the membrane potential could become less negative than the resting potential. A masking effect of facilitated depolarizing potentials on IPSPs was unlikely since IPSPs accompanied by little or no depolarizing potential were also depressed by SR trains. The 75% reduction in IPSP conductance found after repetitive stimulation confirmed that an overlapping conductance was not responsible for the depression of the IPSP. The GABA-induced conductance increase was not depressed by identical trains. Trains of stimulation induced depolarizing shifts in equilibrium potentials for the IPSP (EIPSP) and GABA (EGABA) of approximately 10 mV. These shifts were always greater after SR trains than after ALV trains. Simultaneous recordings of membrane potential and extracellular potassium concentration ([K+]o) with K+-sensitive microelectrodes revealed a direct correlation between the two during a stimulus train. Membrane potential depolarized as much as 18 mV from the peak of the IPSP and [K+]o could increase to a maximum of 10 mM during some trains. A depressant effect (of approximately 50%) of K+ on IPSPs was demonstrated by brief pressure ejection of K+ near the soma. We conclude that repetitive stimulation depresses gIPSP and shifts EIPSP in the depolarizing direction. Whereas gIPSP began to decline after a single conditioning pulse, the additional depression of IPSPs produced by stimulus trains was due in large part to shifts in EIPSP. Depression of gIPSP was not due to desensitization or block of ionic conductances, since gGABA was not reduced. The EIPSP may change as a result of increases in [K+]o. PMID- 2984353 TI - Analysis of decreased conductance serotonergic response in Aplysia ink motor neurons. AB - Micropressure ejection of serotonin (5-hydroxytryptamine, 5-HT) produced excitatory responses in the L14 ink motor neurons of Aplysia that depended on the site of application. Ejection of 5-HT onto the cell body produced a slow response that showed variability in voltage sensitivity between preparations. In contrast, ejection of 5-HT onto the neuropil underneath the cell body produced a response whose amplitude was consistently a linear function of the holding potential, reversing near the predicted potassium equilibrium potential. Subsequent analyses focused on this second response. The neuropil response induced by 5-HT had a linear current-voltage relationship (reversing at ca. -80 mV), was associated with a decrease in input conductance, and was sensitive to changes in the concentration of extracellular K+. Serotonin application in artificial seawater (ASW) containing 30 mM K+ produced a response that reversed close to the altered Nernst potential for K+. The 5-HT response did not appear to be due to secondary activation of interneurons or to depend primarily on extracellular Ca2+, since ejection of 5-HT onto cells bathed in ASW containing 30 mM Co2+ produced responses comparable to, although somewhat attenuated from, those observed in ASW. Serotonin responses similar to those produced in ASW were obtained after perfusing the ganglion with ASW containing Co2+, 4-aminopyridine (4-AP), and tetraethylammonium (TEA). This suggests that the 5-HT-sensitive current is separate from the Ca2+-activated, fast, and delayed rectifying K+ currents. The 5 HT response appeared to be mediated by changes in levels of cAMP. Bath application of the phosphodiesterase inhibitors IBMX (3-isobutyl-1 methylxanthine) or Ro 20-1724, or the adenylate cyclase activator forskolin mimicked the 5-HT response by producing a slow inward current associated with a decrease in membrane conductance. Alteration of cellular cAMP metabolism modulated the response to 5-HT. Exposure of the ganglion to low concentrations of either Ro 20-1724 or forskolin potentiated the 5-HT response. Higher concentrations of these agents largely blocked the response to subsequent 5-HT applications. Bath application of the 8-bromo derivative of either cAMP or cGMP produced a slow inward current associated with a decrease in membrane conductance in cells voltage clamped at the resting potential. Responses to 5-HT were blocked, however, after exposure to 8-bromo-cAMP, but not to 8-bromo-cGMP. These results suggest that 5-HT produces a voltage-independent decrease in a steady state potassium conductance that may be mediated by cAMP.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984354 TI - Comparable patterns of muscle facilitation evoked by individual corticomotoneuronal (CM) cells and by single intracortical microstimuli in primates: evidence for functional groups of CM cells. AB - We compared the averaged responses of forelimb muscles to action potentials of single motor cortex cells and to single intracortical microstimuli (S-ICMS). Activity of precentral neurons and 12 identified forelimb muscles (6 flexors and 6 extensors of wrist and fingers) was recorded in macaques while they performed alternating ramp-and-hold wrist movements. Action potentials of cells that covaried reliably with wrist flexion or extension were used to compile spike triggered averages (spike-TAs) of rectified electromyographic (EMG) activity of six synergistically coactivated muscles. Cells whose spikes were followed by a clear postspike facilitation (PSF) of rectified muscle activity were designated corticomotoneuronal (CM) cells. CM cells typically facilitated a subset of the coactivated muscles called the cell's target muscles. The relative strength of the PSF in different target muscles ranged from clear increases above base-line fluctuations to weak but significant effects. For each CM cell we characterized the "PSF profile" of facilitation across different muscles, defined as the relative strength of PSF in each of the coactivated agonist muscles. After identifying the CM cell's target muscles, we delivered S-ICMS through the microelectrode at the same site. Biphasic stimuli were delivered during the same wrist movements in which the recorded CM cell had been active. Stimulus intensities were too weak (typically 5-10 microA) and their repetition rate too slow (5-15 Hz) to evoke muscle excitation evident in the raw EMG record. However, stimulus-triggered averages (stimulus-TAs) of the rectified EMGs of coactivated muscles revealed consistent patterns of poststimulus facilitation (PStimF). In most cases the muscles facilitated by the CM cell in spike-TAs (n = 60) were also facilitated by S-ICMS in stimulus-TAs. At sites of CM cells the threshold stimulus intensities for evoking a statistically significant effect were between 0.5 and 2 microA. S-ICMS of 5 microA evoked PStimF that was, on the average, six times stronger than the PSF of the CM cell. The height of the facilitation peak relative to base-line fluctuations was 5-60 times greater for the stimuli than the spikes of the CM cell. The average onset latency of PStimF (8.0 +/- 1.2 ms) was 1.3 ms longer than the mean latency of PSF (6.7 +/- 1.4 ms). At two-thirds of the cortical sites where both spike- and stimulus-TAs were computed (n = 30), the PStimF profile exactly matched the PSF profile.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984355 TI - Patterns of facilitation and suppression of antagonist forelimb muscles from motor cortex sites in the awake monkey. AB - Patterns of excitatory and inhibitory effects were produced in antagonistic forelimb muscles by single intracortical microstimuli (S-ICMS) applied to motor cortex sites in macaque monkeys performing ramp-and-hold wrist movements. Stimulus-triggered averages (stimulus-TAs) of rectified electromyographic (EMG) activity revealed poststimulus facilitation and/or suppression in identified flexor and extensor muscles of the wrist and fingers. At 22 cortical sites the action potentials of single cells were also recorded and used to compute spike triggered averages (spike-TAs) of covarying muscles. The set of muscles activated during the movement in which the cell was active are referred to here as "agonists"; those muscles active during wrist movement in the opposite direction are called "antagonists." (At sites where cells were not isolated the muscles showing poststimulus facilitation were called agonists.) Poststimulus effects in agonist muscles typically consisted of facilitation in a subset of the agonists. For 48 sites from which poststimulus effects were tested on both flexors and extensors, the following combinations of effects were observed: 1) pure facilitation of agonist muscles with no effect on antagonists; 2) facilitation of both agonists and antagonists; 3) facilitation of agonist muscles with reciprocal suppression of antagonists; 4) "mixed" facilitation and suppression of synergist muscles; and 5) pure suppression of some muscles with no effect on their antagonists. The suppression effects appeared most commonly in flexor muscles; conversely, facilitation was generally stronger in extensors. Cortical sites eliciting pure suppression of flexor muscles with no facilitation of extensor muscles were found in two monkeys. These purely suppressive effects were observed not only in stimulus-TAs but also in spike-TAs computed from single cells at these sites. Some of these cells increased their activity during wrist extension (but had no detectable effect on the extensor muscles); others discharged during flexion. Several observations suggest that the cortically evoked suppression is mediated by polysynaptic relays. The mean onset latency of the postspike suppression (7.4 ms) produced by inhibitory cells was longer than the mean onset latency of postspike facilitation (6.7 ms) produced by CM cells. Similarly, the mean onset latency of poststimulus suppression (8.9 ms) was longer than that of poststimulus facilitation (8.0 ms). Moreover, suppression was usually weaker than facilitation in the spike-TAs, as well as in stimulus-TAs compiled for the same stimulus intensity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984356 TI - Physiology of a bidirectional, excitatory, chemical synapse. AB - Neurons of the motor nerve net of the jellyfish Cyanea are connected by chemical synapses that, from their ultrastructure, appear to be bidirectional chemical synapses. These synapses were examined physiologically, by recording intracellularly from synaptically connected cells, with the whole cell configuration of the patch-clamp recording technique. Subthreshold depolarizations produced neither small voltage responses indicative of electrical coupling, nor unitary depolarizations suggestive of excitatory postsynaptic potentials (EPSP). Synaptic transmission was affected only when the presynaptic cell was depolarized above spike threshold. The synaptic delay was slightly less than 1 ms at room temperature. The postsynaptic response was initially suprathreshold, resulting in an action potential, but with time this gave way to a large 60 mV amplitude EPSP that did not produce action potentials. The amplitude of the EPSP was directly related to the postsynaptic membrane potential and extrapolated to a reversal potential close to zero mV. Reversal of the EPSP was never observed, even in the presence of intracellular tetrathylammonium (TEA). The relationship between presynaptic depolarization and postsynaptic response was difficult to examine in normal conditions, but in the presence of extracellular lidocaine, which blocked the Na+ and K+ channels in these membranes, a distinct relationship was apparent. The synapse was physiologically nonpolarized and conducted equally well in either direction with a constant synaptic delay. PMID- 2984358 TI - Development of the ipsilateral retinothalamic projection in the frog Xenopus laevis. I. Retinal distribution of ipsilaterally projecting cells in normal and experimentally manipulated frogs. AB - The distribution of ipsilaterally and contralaterally projecting cells within the retina in Xenopus laevis was studied by injection of horseradish peroxidase into the thalamus on one side of the brain and subsequent determination of the locations of retrogradely labeled cells in both retinas. In normal animals, contralaterally projecting cells were found throughout the retina. Ipsilaterally projecting cells, in contrast, were most frequent in temporoventral retina and largely absent from dorsonasal retina as well as from a region surrounding the nerve head. A similarly restricted distribution of ipsilaterally projecting cells was observed in retinas of animals after regeneration of one optic nerve as well as in animals from which one eye was removed prior to the time when the ipsilateral projection first develops. The restricted distribution of ipsilaterally projecting cells in normal animals raises the possibility that these cells may be produced relatively late in development. This hypothesis is explored in the following paper (Hoskins, S.G., and P. Grobstein (1985) J. Neurosci. 5: 920-929). The fact that similar distributions were seen in normal and experimental animals implies that organization of the ipsilateral retinothalamic projection in X. laevis is not critically dependent either on particular patterns of axonal organization which may be present during normal development or on interactions among fibers from the two eyes. PMID- 2984357 TI - Localization of type I benzodiazepine receptors to postsynaptic densities in bovine brain. AB - The subcellular localization of central-type benzodiazepine receptors in bovine cerebral cortex, cerebellum, hippocampus, and corpus striatum has been studied. In all regions except for the corpus striatum, benzodiazepine receptors are most highly enriched in purified postsynaptic densities (PSDs) prepared by Triton X 100/hypotonic lysis of purified synaptosomal plasma membranes. Benzodiazepine receptor enrichment in PSDs varies regionally, following the order cerebellum (approximately 8.5-fold enriched relative to crude P2 membranes) greater than cerebral cortex greater than hippocampus greater than striatum (no significant enrichment); the percentage of putative type I benzodiazepine receptors in each of these brain regions follows the same rank order. In cerebral cortex, analysis of displacement of the benzodiazepine antagonist [3H]Ro-15-1788 by the type I selective drug CL-218,872 reveals that PSDs contain type I benzodiazepine receptors exclusively; other subcellular fractions contain mixtures of type I and type II benzodiazepine receptors. Benzodiazepine receptors in PSDs resist further extraction with detergent but can be solubilized with detergent containing greater than or equal to 0.2 M NaCl. The enrichment of detergent resistant/detergent-plus-salt extractable type I benzodiazepine receptors in PSDs might account in part for the differential solubilization of type I and type II benzodiazepine receptors from crude brain membranes previously reported. The benzodiazepine-binding protein in cerebral cortical PSDs was identified by photoaffinity labeling with [3H]flunitrazepam followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. The PSD benzodiazepine-binding protein is identical in molecular weight to the binding protein from whole brain; partial tryptic and alpha-chymotryptic fingerprints are also very similar in PSDs and whole brain. PMID- 2984359 TI - Development of the ipsilateral retinothalamic projection in the frog Xenopus laevis. II. Ingrowth of optic nerve fibers and production of ipsilaterally projecting retinal ganglion cells. AB - We have studied the development of the ipsilateral retinothalamic projection in the frog Xenopus laevis by analyzing patterns of histochemical reaction product resulting from anterograde transport of horseradish peroxidase (HRP) applied to cut optic nerves in animals of various ages. We have also determined the stages during which ipsilaterally projecting ganglion cells are born using a combination of [3H] thymidine autoradiography and retrograde marking of ganglion cells following injection of HRP into the thalamus. Projections to ipsilateral thalamic terminal zones were first detectable beginning at about larval stage 54. There was a clear asynchrony in innervation, with projections to some terminal zones appearing before projections to others; projections to all terminal zones were present by late metamorphic stages. Within individual terminal zones there were progressive increases in the density of the projections as well as changes in their distribution. By these criteria, development of the ipsilateral projection was not complete at the end of metamorphosis but continued for some months thereafter. Our birth dating studies show that ipsilaterally projecting cells are produced relatively late in development and that, like the development of the projection, the production of ipsilaterally projecting ganglion cells continues postmetamorphically. The vast majority of ipsilaterally projecting cells are born over a period beginning at stage 54/55, when the projection first appears. This stage is significant, since it is at approximately this time that thyroxine dependent metamorphic events begin. In the following paper (Hoskins, S.G., and P. Grobstein (1985) J. Neurosci. 5: 930-940) we report studies on the involvement of thyroxine in the development of the ipsilateral retinothalamic projection. PMID- 2984360 TI - Development of the ipsilateral retinothalamic projection in the frog Xenopus laevis. III. The role of thyroxine. AB - The ipsilateral retinothalamic projection in Xenopus laevis normally first appears at about stage 54, at a time when a number of other changes known to be dependent on a rise in circulating levels of thyroxine begin to occur. We have investigated the role of thyroxine in the development of the ipsilateral retinothalamic projection by studying retinal projections and patterns of retinal histogenesis in tadpoles whose ability to produce thyroxine was blocked by treatment with propylthiouracil (PTU), and in similar tadpoles in which thyroxine was restored by injection of small amounts of thyroxine into one eye. PTU-reared tadpoles continue to grow and to add neurons to the retina in a symmetric pattern like that of normal tadpoles at early developmental stages. The PTU-reared tadpoles remained by external criteria at stage 54. Like normal stage 54 tadpoles, the PTU-reared tadpoles either lacked an ipsilateral projection entirely or had an extremely sparse projection. Injection of thyroxine into one eye of PTU-reared tadpoles resulted in the production of substantial ipsilateral projections from the treated eyes as well as shifts to the asymmetric pattern of retinal cell addition which normally begins after stage 54. Such changes were much more prominent in hormone-treated than in untreated eyes, suggesting that they are caused by local action of thyroxine on the treated eyes. With low doses, thyroxine-induced effects on the development of the ipsilateral projection and on retinal histogenesis were restricted to the treated eye. These results suggest that the presence of thyroxine in one eye alone is sufficient to cause the development of the ipsilateral projection. PMID- 2984361 TI - Suppression of hepatic hematopoiesis with radioactive gold (198Au). AB - A patient with idiopathic myelofibrosis of some 20 yr duration developed esophageal varices and ascites. No explanation for increased portal pressure other than hepatic hematopoiesis was found. Consequently, a trial of cobalt irradiation to the liver was undertaken with definite but transient decrease in ascites. Subsequently, two courses of radioactive colloidal gold were given, again with definite but transient beneficial effects on the degree of ascites. This latter benefit occurred without suppression of marrow function. PMID- 2984362 TI - Adverse allergic reaction to technetium-99m methylene diphosphonate. AB - Adverse allergic reactions to radiopharmaceuticals are rare but have been documented in the literature. This report presents data consistent with a definite adverse reaction to the radiopharmaceutical [99mTc]MDP. PMID- 2984363 TI - Uptake of technetium-99m methylene diphosphonate by fractured and osteoporotic bone after a pulse dose of vitamin D3. AB - The effect of a pulse dose of Vitamin D3 on uptake of [99mTc]MDP by fractured and osteoporotic bone, respectively, was compared with D3's effect on uptake by normal bone in rats. At 4, 7, and 14 days, respectively, after femoral fracture, basal uptake was significantly (p less than 0.005) increased at the fracture site by 336.8, 276.1, and 183.5%, respectively, over the contralateral control site. D3-treated rats had lower uptakes than untreated controls at all three fracture sites and at 12 of 15 normal bone sites but analysis of variance showed the uptake differences were not significant. Cortisone-induced osteoporosis caused a significant (p less than 0.05) decrease in basal uptake. The decrease occurred in all nine bone areas studied. D3 caused a significant (p less than 0.05) increase (mean 16.2%) in uptake by these osteoporotic bones, but a significant (0.1 greater than p greater than 0.05) decrease (mean 13.0%) in uptake by the same bones in normal controls. Thus, D3 had an effect on uptake by the bone lesion, osteoporosis, that differed from D3's effect on uptake by fracture or normal bone. PMID- 2984364 TI - The buildup factor: effect of scatter on absolute volume determination. AB - We have developed a new method for generating attenuation-corrected images for use in absolute volume and activity measurements. The technique relies on the use of a set of measured buildup factors to correct for the effects of scatter inherent in the broad-beam conditions of clinical nuclear medicine and requires anterior and posterior count-rate measurements. The scatter correction requires that the well-known attenuation factor e-mud be replaced by 1-(1-e mud)B(infinity), where B(infinity) is the buildup factor at infinite depth. The buildup factors for four different scintillation camera window settings and three different source sizes are reported. The method was validated by calculating phantom volumes and comparing the results to a previously reported technique which does not account for the scatter contribution by assuming mu = 0.15 cm-1. The results showed that the buildup factor method provides less than 7.3% error for volume determinations at all investigated depths, window settings, and source sizes, whereas errors of 3.3-26.7% were found with the other technique. PMID- 2984365 TI - The effect of social support on educational outcomes. AB - A review of the social support literature identified the theoretical framework for this prospective descriptive correlational study. The relationship of social support to educational outcomes was investigated in a convenience sample of 67 junior and 60 senior nursing students. Social support was measured by the Inventory of Socially Supportive Behavior (ISSB), indicating the frequency with which respondents were recipients of supportive actions from their total support network. Outcomes included the cumulative grade point averages of all participants and the NCLEX scores of graduating seniors. Social Support was also examined in relation to self-esteem, measured by the Coopersmith Self-Concept scale and test anxiety, measured by the Spielberger Test Anxiety Inventory. Hypotheses tested were: (1) There is a positive relationship between social support and both cumulative grade point average and NCLEX scores. (2) There is a positive relationship between social support and self-esteem. (3) Social support will have greater influence on outcomes under conditions of stress, defined as high test anxiety. Hypotheses 1 and 3 were not supported. Hypothesis 2 was supported at the .05 significance level, using a Pearson Correlation Coefficient. Additional information included a significant inverse correlation for test anxiety and both grade point average (p = .01) and NCLEX results (p = .05). Implications for educational experiences and future research are discussed. PMID- 2984366 TI - The evolution of a baccalaureate program for registered nurses. AB - Many collegiate schools of nursing are attempting to meet the needs of ever increasing numbers of RNs who are returning to school for the purpose of earning the baccalaureate degree. As non-traditional learners their professional and personal needs vary. Mechanisms have been developed and evaluated to assist the assessment of prior knowledge and skills. Support systems and program changes have been put in place to facilitate accomplishment of the goals of the RN student. The authors discuss ten years' experience with one RN-BSN program, and describe procedures for advance placement credit and special RN courses. Descriptive data and success measures of the 198 graduates of the original curriculum are presented as well as support systems and program changes that proved helpful. More than half of the graduates assumed positions with managerial responsibility. They were more active in professional organizations and nearly half had completed or were enrolled in graduate study. Nearly two-thirds responded positively about their derived benefits of the program listing specific courses, self-confidence, and improvement in critical thinking as some of the gains. Many noted an increased awareness of the need for nurses to work collectively in the pursuit of professional issues. This potential for development of strong leadership from this group in nursing is seen as an asset to our profession. PMID- 2984367 TI - Cognitive style and the behavioral differences of nursing students in the clinical setting. AB - This study examined the applicability of the naturalistic method of ecological psychology to the study of student nurse behavior in a hospital setting in relation to cognitive style. Naturalistic data were collected during a three-hour clinical experience of 12 students selected from a baccalaureate, medical surgical class of 62 junior level students differing on field independence and field dependence as determined by scores on the Group Embedded Figures Test. Field independent and field dependent students were found to differ significantly in some of the following ways: frequency and duration of interactions with the instructor and with the patient, tendency to initiate behavioral units, and type of activities performed during the clinical experience. The naturalistic method was found to be a fruitful approach for the study of student nurse behavior. PMID- 2984368 TI - Self-identified stressors in the role of nursing faculty. AB - A card-sort methodology was found to be a rapid and systematic approach for the identification of stressors for a baccalaureate nursing faculty. It would seem that multiple stressors exist within the role of nursing faculty. These may be categorized into four main sections: stressors related to Academia, Administration, Clinical and Classroom. The methodology provided assistance with prioritizing such stressors for problem solving by the faculty. An issue in conducting this type of research is confidentiality as demonstrated in faculty reluctance to reveal demographic data that might serve to connect responses with the source. PMID- 2984369 TI - Can the generic curriculum function for the returning RN student? PMID- 2984370 TI - Effects of computer-assisted instruction on nursing student learning and attitude. PMID- 2984371 TI - The clinical contract--an approach to competency-based clinical learning and evaluation. PMID- 2984372 TI - Role issues and the nurse educator. PMID- 2984373 TI - Integrating gerontology into the nursing curriculum: process and content. PMID- 2984374 TI - Locus of decision making in schools of nursing. PMID- 2984375 TI - Preparing faculty to teach nursing research. PMID- 2984376 TI - Systematic faculty evaluation: a growing critical concern. PMID- 2984377 TI - Legal implications of academic dismissal and educational malpractice for nursing faculty. AB - Recent legal decisions affect nursing education not only in the area of academic dismissal but also in the area of educational malpractice. Nursing faculty may be caught in an interesting "catch-22" phenomenon. There exists a possibility where an educator may be sued for both failing, and not failing, a student. An incompetent graduate may sue for inadequate education, and a dismissed student may sue the faculty member for failing to treat the student in a logical and reasonable manner. Although courts have recognized that faculty members are uniquely qualified to observe and judge all aspects of their students' academic performance, nursing instructors have a double responsibility; one to the client and the other to the student. Clinical nursing faculties must meet a standard of care with respect to the client and a standard of conduct with respect to the student. PMID- 2984378 TI - The client base of nursing, 1920-1930. AB - History should serve two functions: it should provide a more complete picture of the development of civilization, and it should make us aware of our beginnings, where we stand today, and the direction in which we are heading. People are the dynamic element in history. This study provides a historical perspective of nursing through an investigation of social attitudes, economic status and educational opportunities in New England from 1920-1930. Theories from Clarke, Shryock and Baldridge were utilized. An analysis of historical data suggests that nursing educators need to address the current climate of opinion concerning nursing, to analyze the market place to determine the type nursing leaders needed to guide the future of nursing, and to re-evaluate the client-base of professional nursing with a view to attracting and retaining highly qualified people capable of achieving positive collegial relations with others while improving nursing autonomy. PMID- 2984379 TI - Stress inoculation: a method of helping students cope with anxiety related to clinical performance. PMID- 2984380 TI - Nursing care analysis: a tool to develop problem solving. PMID- 2984381 TI - Ambulatory pediatric screening: a valuable learning experience for nursing students. PMID- 2984382 TI - Bringing baccalaureate nursing education to the rural setting. PMID- 2984383 TI - Therapeutic swimming: a different kind of nursing. PMID- 2984384 TI - Nursing students' impact on a system of care for the elderly. PMID- 2984385 TI - Assimilating the learning needs of RN students into the clinical practicum. PMID- 2984386 TI - Are complex multiple-choice options more difficult and discriminating than conventional multiple-choice options? AB - Differences in item difficulty between conventional and complex multiple-choice (M-C) items measuring objectives written at the six levels of the cognitive domain and three content areas within a basic health professions subject matter area were investigated. Conventional and complex M-C items were also compared in terms of item discrimination. Five hundred fifteen senior baccalaureate nursing students randomly responded to one of two packages of 18 one-item pharmacology achievement tests. Items in the two packages were matched in all respects except for the type of option. A 2 X 3 X 6 repeated measures ANCOVA with method of pharmacology as the covariate showed a statistically significant (p less than .01) interaction of option X content X cognitive level. Scheffe tests revealed that complex items were more difficult than conventional option items at the knowledge level only across all content areas investigated. No statistically significant differences were found between the two option types discriminating ability. Results of this research question the validity of the assumption that complex items are more difficult and discriminating than conventional M-C items. PMID- 2984387 TI - Professional nursing education: what is its purpose? AB - Rogers (1970, p. 88) has written "The purpose of professional education is to provide the knowledge and tools whereby an individual may become an artist in his field. It is not to prepare the skilled practitioner." This statement is used to provide a basis for discussion of some issues involved in relation to nursing and nursing education. The problem area is first clarified before discussion moves on to show the relevance of conceptual models and theory to professional nursing education and to look at the kinds of knowledge and tools which could be provided to promote artistry in nursing. PMID- 2984388 TI - Evaluation of a subcutaneously implanted hydroxylapatite-avitene mixture in rabbits. AB - Avitene, a microfibrillar collagen hemostatic agent was mixed with hydroxylapatite and implanted subcutaneously along the inferior border of the mandible in ten rabbits. This mixture clearly enhanced surgical manipulation of the implants. Radiographic and histologic examination of the implant sites at one, three, and six months showed an acceptable host response with no apparent alteration of the favorable characteristics of hydroxylapatite implants. PMID- 2984389 TI - Plasmacytoid myoepithelioma of a minor salivary gland. AB - Plasmacytoid myoepitheliomas are rare salivary gland neoplasms that probably represent a variant of the pleomorphic adenoma. They appear to occur more frequently in the minor salivary glands of the mouth. The mean average age at time of diagnosis of intraoral lesions is 18.8 years. Cytologic pleomorphism is a frequent histologic feature, which has led to a questionable diagnosis of malignancy in five out of ten cases. Caution is advocated in the evaluation of a salivary gland neoplasm with such features, and confusion with primary squamous cell or undifferentiated carcinoma of the salivary glands should be avoided. Patients who have cytologically pleomorphic lesions should be assessed for evidence of metastases. Rapid growth, neurologic evidence of sensory or motor dysfunction, and bone invasion should be regarded as ominous clinical parameters. Therapy should be directed toward complete surgical extirpation. PMID- 2984390 TI - Steindler lecture. Binding sites in fetal and growth plate cartilage. AB - In addition to genetic and nutritional factors, linear growth during the prenatal and postnatal periods is controlled by peptide, steroid, and thyroid hormones interacting with the receptors present on the membrane or in the cytosol and nuclei of growth plate cartilage. Using standard procedures, insulin and "nonsuppressible insulin-like activity" (a somatomedin) showed significant binding in 600, 15,000, and 105,000 g membrane fractions of epiphyseal cartilage of immature animals. The binding of growth hormone and prolactin was small and probably not significant. Specific uptake of glucocorticoid was demonstrated in viable canine chondrocytes, but not of androgen, estrogen, or vitamin D3 metabolite. A triiodothyronine receptor was present in nuclei from dog epiphyseal cartilage. Hormones that lack binding may affect cartilage only indirectly. Hormone receptors were studied in those portions of fetal growth cartilage that will later evolve into an ossification center, articular cartilage, and epiphyseal cartilage. Cytosol fractions contained a receptor for glucocorticoid but not for androgen or estrogen. Zonal analysis showed a higher level in the peripheral and central sections than in the palisade section. Triiodothyronine binding was also detected in nuclei prepared from whole fetal cartilage. Heterogeneity of cell function was obvious in fetal cartilage. Cell division was high in the central and peripheral zones as well as the upper half of the palisade zone, but low in the lower palisade section. Proline and sulfate incorporation predominated in the palisade section compared with the central and peripheral sections. Disease states with changes of metabolic activities in the cartilage may perhaps be better understood with a clearer knowledge of receptor levels and interactions. PMID- 2984391 TI - Increased collagenolytic activity in severed and sutured tendons following topical application of exogenous collagen in chickens. AB - To verify the role of collagenase in reduction of peritendinous adhesion by topical application of exogenous collagen, the flexor tendons of 30 chickens were severed and sutured. Exogenous, native enriched collagen solution (ECS) was introduced in the tendon sheath via a polyethylene catheter. The effect of ECS on collagenolytic activity in the healing tendon was assessed 1, 2, and 3 weeks later both by determining the relative amounts of dialyzable protein and hydroxyproline and by using the collagen film collagenase assay. The results obtained indicated a significant increase in both dialyzable hydroxyproline level and collagenolytic activity in the ECS-treated tendons as compared with the untreated controls. It is suggested that the effect of the topically applied exogenous collagen on increasing the collagenolytic activity may be directly related to previously observed increased gliding capacity of the tendons in the same experimental model. PMID- 2984392 TI - Bone ingrowth into three different porous ceramics implanted into the tibia of rats and rabbits. AB - Three different porous ceramics--calcium aluminate, calcium hydroxyapatite, and tricalcium phosphate--were implanted into the proximal tibia in rats and rabbits to study the interactions between these ceramics and a bony site that is abundant in bone marrow. New bone was consistently formed within the bone marrow surrounding and adjacent to all three types of ceramics. Calcium hydroxyapatite and tricalcium phosphate ceramics permitted bone ingrowth into their pores. The newly formed bone was found only in the part of the ceramic intruding into the bone, not in the portion protruding in the soft tissues outside the tibia. Partial replacement of the new bone opposite the medullary portion by new hemopoietic marrow occurred with longer implantation times. In contrast, no bone was seen within the pores of any of the calcium aluminate implants. PMID- 2984393 TI - Human papillomavirus infection and cancer of the uterine cervix. PMID- 2984394 TI - Induction of nephroblastoma-like renal tumours in rats by a single dose of 1,2 dimethylhydrazine. AB - A single injection of 1,2-dimethylhydrazine (200 mg/kg s.c.) resulted in the development of renal tumours in 70 per cent of treated rats within 1 year. The tumours were frequently bilateral (36 per cent) and grew to a large size (up to 100 g) before causing death. Lung metastases were found in one case. Histologically, the majority of the tumours (73 per cent) exhibited the classical triphasic morphological features of nephroblastoma (Wilms' tumour) with varying proportions of epithelial (glomeruloid and tubular structures), stromal (fibrous tissue, smooth muscle, cartilage) and blastematous elements. This pathological resemblance to human nephroblastoma suggests the possibility of using dimethylhydrazine-induced renal tumours as a model of Wilms' tumour. PMID- 2984395 TI - Pseudohermaphroditism, glomerulopathy, and Wilms tumor (Drash syndrome): frequency in end-stage renal failure. AB - The sporadic concurrence of male pseudohermaphroditism and chronic glomerulopathy is associated with an extremely high risk of Wilms tumor. We report our experience with an infant who developed this triad (Drash syndrome) and review the 21 patients described in the literature, to emphasize the importance of early diagnosis and to suggest guidelines for management. The dysgenetic gonads are always intra-abdominal and carry a 20% to 30% risk for malignancy. The external genitalia are frequently ambiguous (77%); some children are phenotypically normal females. The glomerulopathy typically leads to end-stage renal failure in infancy; the subsequent death rate has, to date, been 68%. The clinical presentation of renal disease is variable and includes congenital nephrotic syndrome (14%) and infantile nephrotic syndrome (41%); 27% of patients develop proteinuria and renal insufficiency between the ages of 1 and 3 years. The high risk of Wilms tumor (55% in this review) mandates regular tumor surveillance, and prophylactic bilateral nephrectomy and gonadectomy once irreversible renal failure develops. PMID- 2984396 TI - Changing concepts: liver replacement for hereditary tyrosinemia and hepatoma. PMID- 2984398 TI - Survival of cytomegalovirus on environmental surfaces. PMID- 2984397 TI - Normal macrophage function in infants receiving Intralipid by low-dose intermittent administration. AB - The effect of soybean oil emulsion (Intralipid) therapy on serum complement levels was determined in infants who received Intralipid therapeutically (1 gm/kg over 12 hours, every other day). The effect of Intralipid on macrophage priming for increased superoxide anion production was studied in a mouse model. Intralipid administration did not affect either macrophage function. Serum levels of C2 and C4, complement components synthesized and secreted exclusively in macrophages, were not decreased either during the week the infants received Intralipid or in the week following administration. Macrophages from mice that had received Intralipid produced similar amounts of superoxide anion, as did macrophages from mice that had received saline solution. Our data suggest that macrophages in infants receiving Intralipid in this regimen will function normally. PMID- 2984399 TI - Renal toxicity of cisplatin in children. AB - We measured renal function in 22 children receiving cisplatin as initial treatment for neuroblastoma or malignant germ cell tumors. Glomerular filtration rates were estimated from the plasma clearance of 51Cr-EDTA and were compared with measurements of plasma creatinine concentration and creatinine clearance. The degree of cisplatin-induced renal damage varied widely, and plasma creatinine measurements and creatinine clearances were not reliable guides to glomerular filtration rate. Renal function in children receiving cisplatin should be monitored by measurement of glomerular filtration rate with an isotope clearance technique. PMID- 2984400 TI - Factors influencing excystation in Cryptosporidium oocysts from cattle. PMID- 2984401 TI - Developmental physiology of cestodes: cyclic nucleotides and the identity of putative crowding factors in Hymenolepis diminuta. AB - Worm-conditioned saline (WCS) was prepared by incubating Hymenolepis diminuta from crowded infections for 12 hr in a balanced salt solution. The effect of the WCS on the incorporation of [3H] thymidine into DNA in the anterior regions of fresh H. diminuta was compared to effects produced by the cyclic nucleotides in the WCS. Cyclic AMP and cGMP were found in the WCS, and cGMP but not cAMP (at the concentration in WCS) caused some inhibition of DNA synthesis. For further study of the effects of cyclic nucleotides, worms were incubated with theophylline, caffeine, 3-isobutyl-1-methyl xanthine, 2-deoxy cGMP, and L-ascorbic acid, all of which produced some inhibition of [3H] thymidine incorporation. Treatment of WCS with 3',5' cyclic nucleotide phosphodiesterase abolished part of its inhibitory activity, i.e., that part presumed to be due to cGMP. When worms were incubated in the presence of succinate, acetate, D-glucosaminic acid, and cGMP simultaneously and in the concentrations each was found in the WCS, DNA synthesis was inhibited to a degree equal to that found in the WCS. Thus these substances apparently represent the putative crowding factors in the WCS. WCS prepared with worms from different population densities contained the same levels of cAMP but varied in content of cGMP, which decreased as the worm density increased. WCS prepared with patent worms contained high levels of cAMP, but the same amounts of cGMP as WCS prepared with 10-day-old worms. At least some inhibitors of cyclic nucleotide phosphodiesterase inhibited the secretion of cGMP by the worms. Levels of cGMP in the host intestine varied with the presence or absence of worms, number of worms, and area of the intestine. PMID- 2984402 TI - Intestinal and serum immune response to a naturally acquired rotavirus gastroenteritis in children. AB - Seventeen children (mean age: 2.0 years, range: 36 days-8 years) hospitalized with acute gastroenteritis were investigated. Thirteen children had a rotavirus infection while four did not. Rotavirus serum IgA as well as ScIg, i.e., antirotavirus immunoglobulin containing secretory component, increased rapidly after rotavirus infection. While rotavirus IgA persisted in serum for at least 6 months, rotavirus ScIg disappeared from serum in less than 4 months. Rotavirus IgG could be detected in serum during the early stage of the infection and was still high after 6 months. The patients with nonrotavirus acute gastroenteritis did not show any of the above-mentioned serological hallmarks of those with rotavirus infection. The amounts of rotavirus ScIg found in serum about 1 week after the infection correlated to the amounts of rotavirus ScIg in duodenal fluid. Six months after the infection, rotavirus IgA was found in the feces of the majority of the patients while rotavirus ScIg could be detected only in one patient. The amounts of rotavirus IgA in sera and intestinal secretions showed identical patterns in the acute phase of the disease as well as after recovery. The same applied to rotavirus ScIg. These findings could be useful in future evaluations of vaccines and immunity against rotavirus infections. PMID- 2984403 TI - Hydroxylapatite as an alloplastic graft in the treatment of human periodontal osseous defects. AB - Twelve patients, 32 to 60 years of age, received a polycrystalline ceramic form of pure dense hydroxylapatite as an alloplastic bone implant material in intrabony defects following reflection of full mucoperiosteal flaps, root planing and defect-curettement. The defects were measured from an acrylic stent, using an endodontic silver point which was placed to the base of the defect. Similarly, debrided and curetted defects in the same patients were not implanted and served as controls. Recalls for documentation and plaque control were at 1, 2, and 4 weeks, and at 3, 6 and 9 months. Measurements relating to changes in defect-depth were made upon reentry at 9 months. The twelve defects, serving as controls, showed very little difference between the pretherapy and 9-month measurements. The initial mean measurement from the base of the defect to the highest alveolar crest was 4.27 mm and the 9-month mean measurement after curettage only was 3.36 mm. In terms of resolution of the original defect this amounted to 19.49% reduction, but a 0.46-mm mean loss in height of the alveolar crest provided an actual percentage fill of the original defect of 9.91%. Of sixteen experimental defects, the same initial mean measurement from the base of the defect to the highest alveolar crest was 5.18 mm and the 9-month mean measurement after grafting was 2.43 mm. In terms of resolution of the original defect, this amounted to a 53.57% reduction, but in contradistinction to the curettage sites, a mean increase in height of the highest alveolar crest of 0.61 mm gave a true percentage fill of the original defect of 66.89%. At the 9-month reentry, the implanted mass seemed to be partially "calcified" and was resistant to penetration with a probe or removal with a curette. The data and clinical impression strongly suggest that hydroxylapatite has a potential as an alloplastic implant with clinically apparent acceptance by the soft and hard tissues. PMID- 2984404 TI - The use of a porous hydroxylapatite implant in periodontal defects. I. Clinical results after six months. AB - Twenty-five patients with advanced periodontal destruction were used in the study. Following initial therapy, two angular interproximal defects were selected in each patient. During flap surgery a porous hydroxylapatite implant shaped to fit the periodontal defect was placed in one defect, the other defect was used as nonimplanted control. The material used for implantation was a hydroxylapatite replicate of coral from the genus Porites, with a pore size of 190 to 220 micron. Clinical parameters were measured prior to flap surgery for each of the defects. An occlusal acrylic stent was used to give a stable reference point for pocket depth, attachment level and gingival margin height measurements. Also gingival fluid, gingival inflammation, plaque index and tooth mobility were recorded. Periapical radiographs using a standardized positioning device were also taken. At the time of surgery, the depth of the osseous defect and the height of the alveolar crest were recorded. After 6 months the clinical measurements were repeated and a re-entry surgery was carried out in 15 selected sites. Results showed that the porous implant produced statistically significant reduction in pocket depth, in the depth of osseous lesion, and a statistically significant gain in attachment level, as compared to control areas. PMID- 2984405 TI - Nutritional polyneuropathy and its manifestations in the lower extremity. PMID- 2984407 TI - Contact dermatitis caused by plaster casting material. PMID- 2984406 TI - Use of Cortisporin Otic Solution in phenol nail surgery. PMID- 2984409 TI - Sepsis and calcium pyrophosphate deposition disease in the same joint. A case report. PMID- 2984408 TI - Nuclear medicine: implications for podiatry. PMID- 2984411 TI - Lipoxygenase inhibitors and cyclic AMP-mediated insulin secretion caused by forskolin, theophylline and dibutyryl cyclic AMP. AB - Forskolin caused a marked and a concentration-dependent elevation of cyclic AMP content in isolated pancreatic islets (EC50, 10 microM). Cyclic AMP level reached a plateau within 30 min after the addition of 10 microM forskolin. In a low glucose (3.3 mM) medium, forskolin induced slight but significant insulin secretion in a concentration-dependent manner (EC50, 0.3 microM). When the glucose concentration was increased to 5.5 mM, marked enhancement of insulin secretion was observed with forskolin (EC50, 0.5 microM). Lipoxygenase inhibitors, such as nordihydroguaiaretic acid, 3-amino-1-(trifluoromethylphenyl) 2-pyrazoline and 1-phenyl-3-pyrazolidinone failed to affect the forskolin stimulated cyclic AMP generation. The selective cyclooxygenase inhibitor indomethacin also had no influence on forskolin-stimulated cyclic AMP generation. Insulinotropic effects of forskolin, however, were suppressed by these lipoxygenase inhibitors but not by indomethacin. Both nordihydroguaiaretic acid and 1-phenyl-3-pyrazolidinone also prevented the insulinotropic effects of theophylline and dibutyryl cyclic AMP, whereas indomethacin failed to inhibit them. It seems conceivable that a lipoxygenase product(s) is involved in the insulin secretory process distal to cyclic AMP generation, or that alternatively a lipoxygenase product(s) is permissively involved in the insulin secretory process independently from the cyclic AMP-mediated process. PMID- 2984410 TI - Effects of pentobarbital on t-[35S]butylbicyclophosphorothionate and [3H]flunitrazepam binding to membrane-bound and solubilized preparations from rat forebrain. AB - The binding sites for t-[35S]butylbicyclophosphorothionate ([35S] TBPS) and [3H]flunitrazepam ([3H]Flu) were solubilized from freshly prepared and washed membranes from rat forebrain with 20 mM 3-[(cholamidopropyl)-dimethylamonio]-1 propanesulfonate in the presence of protease inhibitors. Approximately 64% of the protein, 56% of the [35S]TBPS sites and 45% of the [3H] Flu sites were solubilized. The soluble and membrane-bound binding sites for [3H]Flu were relatively stable on storage at 0 degree C or at -65 degrees C for 11 days. Binding of [35S]TBPS to membranes was increased after freezing the membranes at 65 degrees C for 1 to 11 days and was relatively unchanged if the membranes were stored at 0 degree C for 1 to 11 days. Solubilized [35S]TBPS binding sites were stable when stored at -65 degrees C but declined to 20% of control when stored at 0 degree C for 11 days. Tested shortly after preparation, [35S]TBPS binding to both soluble and membrane preparations demonstrated similar affinity, temperature dependence and noncompetitive inhibition by pentobarbital (pb). Stimulation by pb of [3H]Flu binding to the soluble fraction decayed on storage of the solubilized material at 0 degree C with a time course similar to the loss of [35S]TBPS binding. This finding might suggest that the stimulation of [3H]Flu binding by pb is mediated via its action on TBPS sites. However in membrane preparations stored at 0 degree C, [35S]TBPS binding was stable whereas the pb effects declined with time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984412 TI - H+ gradient-dependent transport of aminocephalosporins in rat renal brush border membrane vesicles: role of H+/organic cation antiport system. AB - The transport of aminocephalosporin antibiotics such as cephalexin, zwitterion, by rat renal brush border membrane vesicles has been studied in relation to the transport system of organic cation. Cephalexin uptake was inhibited by organic cations such as tetraethylammonium, 1N-methylnicotinamide and choline, whereas cefazolin uptake was not. Tetraethylammonium uptake was inhibited by aminocephalosporins and stimulated by the countertransport effect of cephalexin. In agreement with the transport mechanisms of organic cation, the presence of an H+ gradient ([H+]i greater than [H+]o) induced a marked stimulation of aminocephalosporin uptake, and this uptake was inhibited by HgCl2 and tetraethylammonium. These results suggest that aminocephalosporins can share a common carrier transport system with organic cation in renal brush border membranes, and this system could be a part of the mechanisms of tubular secretion for aminocephalosporins. PMID- 2984413 TI - Two distinct populations of [3H]prazosin and [3H]yohimbine binding sites in the plasma membranes of rat mesenteric artery. AB - Postsynaptic alpha adrenoceptor subtypes have been investigated by radioligand binding studies in plasma membrane vesicles prepared from rat mesenteric arteries using [3H]prazosin and [3H]yohimbine. Both the radioligands displayed monophasic saturation in binding with a single component on Scatchard analysis. In the estrogenized female rat mesenteric artery, the specific binding of [3H]prazosin was rapid, saturable, reversible and of high affinity (0.65 +/- 0.05 nM) with a maximum binding capacity (Bmax) of 177 +/- 14 fmol/mg of protein. The maximum number of [3H]yohimbine binding sites were 427 +/- 31 fmol/mg of protein with the Kd equal to 34.5 +/- 3.8 nM. There was no evidence of cooperativity in the binding of both the ligands. The Kd values of [3H]prazosin and [3H]yohimbine, calculated from their respective kinetic analyses of binding, were in good agreement with the Kd values estimated from Scatchard plots. Prazosin was 15,000 times more potent in competing at the [3H]prazosin binding sites than at the [3H]yohimbine sites. In contrast, unlabeled yohimbine was 100-fold more potent in competing at the [3H]yohimbine binding sites than at the [3H]prazosin sites. The affinity of BE 2254 was 10,500 times higher for the [3H]prazosin binding sites than its affinity for the [3H]yohimbine binding sites. Non-alpha adrenoceptor antagonists competed poorly for both the radioligand binding sites. The Kd and Bmax of [3H]prazosin and [3H]yohimbine binding in the membranes of male rat mesenteric arteries were not significantly different from the corresponding values in the membranes of estrogenized female rat mesenteric artery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984414 TI - Muscarine stimulates the hydrolysis of inositol-containing phospholipids in the superior cervical ganglion. AB - Previous studies have shown that muscarine increases the incorporation of 32Pi and [3H]inositol into phosphatidylinositol in the superior cervical ganglion of the rat. Because the first event in agonist-stimulated phospholipid turnover is thought to be the hydrolysis of phosphatidylinositol or of phosphatidylinositol phosphates, we measured the accumulation of [3H]inositol phosphates in ganglia in which these lipids had been labeled by preincubation with [3H]inositol. The production of [3H]inositol phosphates under these conditions presumably reflects the activity of a phospholipase C in the ganglion. Muscarine caused a large increase in the formation of [3H]inositol phosphates. Most of this increase was in the form of [3H]inositol-1-phosphate. The stimulation of [3H]inositol phosphate accumulation by muscarine was not dependent upon the presence of extracellular Ca++. Agents that increase Ca++ influx caused only a small increase in the accumulation of [3H]inositol phosphates. We also measured the formation of [3H]inositol phosphates in extracts of the ganglion. These extracts contained a phospholipase C activity that was stimulated by deoxycholate and that hydrolyzed phosphatidylinositol phosphates more actively than phosphatidylinositol. This phospholipase C activity was Ca++-dependent. We propose that muscarine may activate this phospholipase C in the intact ganglion and that muscarine increases phospholipase C activity by some mechanism other than by increasing the influx of Ca++. PMID- 2984416 TI - Abnormalities in central and peripheral nerve conduction in patients with anorectal incontinence. AB - Thirty women with idiopathic anorectal incontinence were investigated by nerve conduction studies. Twenty-two (73%) patients had abnormal nerve conduction studies. In 7 (23%) of these patients there was conduction delay in the cauda equina between L1 and L4 vertebral levels but in the remainder the distal innervation of the pelvic sphincters was abnormal. PMID- 2984415 TI - Autoradiographic characterization of N-methyl-D-aspartate-, quisqualate- and kainate-sensitive glutamate binding sites. AB - Quantitative autoradiography was used to characterize the pharmacological specificity and anatomical distributions of subtypes of L-[3H]glutamate binding sites in rat brain. One population of sites was sensitive to N-methyl-D-aspartate (NMDA) and other compounds thought to be specific for the NMDA receptor. This site was enriched in stratum radiatum of hippocampus (CA1) where it constituted about 80% of glutamate binding sites and it represented a variable portion of glutamate binding sites throughout the brain. A second population of sites had a high affinity for quisqualate. Approximately 80% of glutamate binding sites in cerebellar molecular layer were of the high affinity quisqualate type. The number of these sites was greatly increased in the presence of Cl- and Ca++ ions. A subset of the high affinity quisqualate sites was sensitive to competition by kainate, particularly in stratum lucidum of hippocampus; the density of these high affinity kainate-sensitive sites was decreased in the presence of Ca++ but not Cl- ions. At high concentrations quisqualate competes for all glutamate binding sites, as reported previously. There was a good correspondence between the density and distribution of low affinity quisqualate sites and NMDA-sensitive sites. Pharmacological analysis suggested that the low affinity quisqualate site and the NMDA site are equivalent. Anatomical and pharmacological evidence suggests that the NMDA-, (high affinity) quisqualate- and kainate-sensitive glutamate binding sites may correspond to the physiologically defined NMDA, quisqualate and kainate receptors. PMID- 2984417 TI - Synthesis and antitumor and antiviral activities of a series of 1-beta-D ribofuranosyl-5-halocytosine (5-halocytidine) cyclic 3',5'-monophosphates. AB - A series of 1-beta-ribofuranosyl-5-halocytosine cyclic 3',5'-monophosphates (1-4) has been prepared. Direct halogenation of cytidine 3',5'-monophosphate (cCMP) yielded the Cl, Br, and I compounds while 5-F-cCMP (1) was obtained on cyclization of the 5'-monophosphate. On in vitro testing of 1-4 against L1210 and P388 leukemias, only 1 showed significant low-level activity (ID50 = 3.1 X 10(-4) mmol/L). Derivatives 2-4 were inactive at 10(-1) mmol/L and also proved to have low viral ratings against a series of RNA and DNA virus strains in vitro. By contrast the 5-F-cCMP showed moderate activity against VV, HSV-1, and HSV-2 strains (VR = 0.6-0.9). Both 5-fluorocytidine and 5-fluorocytidine 5' monophosphate had marked antiviral activity (VR = 1.0-2.1) with the above viruses as well as with parainfluenza virus type 3. The nucleoside and nucleotide also were more active than 5-F-cCMP against L1210 and P388 cells. However, comparison of the cytotoxicities and antiviral ED50 values of 5-F-cCMP, 5-fluorocytidine 5' monophosphate, and 5-fluorocytidine suggests a potential therapeutic advantage for 5-F-cCMP. Possible rationales for these activities are discussed in terms of 5-F-cCMP and the corresponding 5'-monophosphate as potential prodrugs and as sources, following enzymatic deamination, of cytotoxic 5-fluorouridine or its 5' monophosphate. PMID- 2984418 TI - Aldosterone antagonists. 1. Synthesis and biological activities of 11 beta,18 epoxypregnane derivatives. AB - Several steroid derivatives having the 11 beta,18-epoxypregnane skeleton, 7, 8, 19, 20, 21, and 31, were synthesized to evaluate their antialdosterone activity. Among them, 3-(9 alpha-fluoro-17 beta-hydroxy-3-oxoandrost-4-en-17 alpha yl)propionic acid gamma-lactone (31) possessed fairly strong binding affinity for the cytoplasmic mineralocorticoid receptor of rat kidney and exhibited good aldosterone antagonist activity in an in vivo assay. However, its agonistic nature cannot be ignored. The properties of 31 as an aldosterone antagonist were enhanced by its very low to negligible binding affinity for the androgen, progestin, estrogen, and glucocorticoid receptors. PMID- 2984419 TI - Angiotensin-converting enzyme inhibitors: synthesis and biological activity of acyl tripeptide analogues of enalapril. AB - The synthesis and biological activity of a series of inhibitors of angiotensin converting enzyme (EC 3.4.15.1) are described. Incorporation of the substituted N carboxymethyl dipeptide design of enalapril (MK-421) into acyl tripeptides and larger peptides yielded potent inhibitors of the enzyme. These can be viewed as substrate analogues in which the carbonyl of the scissile peptide bond is replaced by a CHCO2H group. Several of the analogues described possess inhibitory potency equal to that of enalaprilat (MK-422), but none achieves an increase in potency which would demonstrate additional binding interactions contributed by the extended peptide chain. Application of the design described may be useful for inhibition of other metallopeptidases. PMID- 2984420 TI - 1,3-Dialkyl-8-(p-sulfophenyl)xanthines: potent water-soluble antagonists for A1- and A2-adenosine receptors. AB - A series of 8-(substituted phenyl) derivatives of theophylline and other 1,3 dialkylxanthines were evaluated for potency and selectivity as antagonists at A1- and A2-adenosine receptors in brain tissue. Theophylline has a similar potency (Ki = 14 microM) at both A1 and A2 receptors. 8-Phenyltheophylline is 25-35-fold more potent as an adenosine receptor antagonist than theophylline, while 8 phenylcaffeine is only 2-3-fold more potent than caffeine. A p-hydroxyaryl substituent enhances the potency of 8-phenyltheophylline as an adenosine antagonist. p-Carboxy- and p-sulfoaryl substituents reduce potency of 8 phenyltheophylline, yielding water-soluble adenosine antagonists, which are some 2-5-fold more potent than theophylline at adenosine receptors. None of the 8 (substituted phenyl)theophyllines are particularly selective as antagonists toward A1- and A2-adenosine receptors. 1,3-Dipropyl-8-phenylxanthine represents a potent and somewhat selective A1-receptor antagonist about 23-fold more potent at A1 receptors than at A2 receptors. A p-hydroxyaryl substituent further enhances potency of the 1,3-dipropyl-8-phenylxanthine at both A1 and A2 receptors. The 8 (2-amino-4-chlorophenyl)-1,3-dipropylxanthine is a very potent and selective antagonist for A1 receptors, being nearly 400-fold more potent at A1 than at A2 receptors. The water-soluble 8-(p-sulfophenyl)- and 8-(p-carboxyphenyl)-1,3 propylxanthines no longer exhibit marked selectivity. Both compounds are much more potent as adenosine antagonists than theophylline. The striking selectivity of 1-isoamyl-3-isobutylxanthine as an A1 antagonist is retained in the 8-phenyl derivative but is virtually lost in the 8-p-sulfophenyl derivative. PMID- 2984422 TI - Specific inhibition of benzodiazepine receptor binding by some N-(indol-3 ylglyoxylyl)amino acid derivatives. AB - Several N-(indol-3-ylglyoxylyl)amino acid derivatives were synthesized and tested for their affinity for the benzodiazepine receptor in bovine cortical membranes. From these compounds, the N-[(5-chloro-, 5-bromo-, or 5-nitroindol-3 yl)glyoxylyl]glycine or -alanine esters were clearly the most potent, while the 5 methoxy analogues were considerably less active. Moreover, esters were more active than the corresponding acids. It is concluded that the affinity of these derivatives for the benzodiazepine receptor is profoundly dependent on amino acid molecular size, as well as the hydrophobic and electronic properties of the compounds. PMID- 2984421 TI - 5-Cinnamoyl-6-aminouracil derivatives as novel anticancer agents. Synthesis, biological evaluation, and structure-activity relationships. AB - A biological evaluation in the series of 5-cinnamoyl-6-aminouracils has been undertaken. These compounds have been found to be in an extended planar conformation fitting well with a possible stacking interaction between the nucleic bases of DNA; thus an eventual anticancer activity by intercalation could be hoped. 1,3-Dimethyl-5-cinnamoyl-6-aminouracil was found to be active when administered ip against ip-implanted P388 leukemia in vivo (percent T/C = 124). Two other compounds, 1,3-dimethyl-5-cinnamoyl-6-[(2-morpholinoethyl)amino]uracil and 1,3-dimethyl-5-cinnamoyl-6-[(2-piperidinoethyl)amino]uracil, bearing a hydrophilic side chain on the 6-amino group, have exhibited cytoxic activity in vitro against L1210 leukemia. Structure-activity relationships have been determined from these results and from studies of biological interactions with DNA. PMID- 2984423 TI - Isolation of St. Louis encephalitis virus from adult Dermacentor variabilis (Acari: Ixodidae). PMID- 2984424 TI - De novo tandem duplication 9p (p12----p24) with normal GALT activity in red cells. AB - A 3 month old boy with a tandem duplication 9p (p12----p24) is reported. Both clinical and dermatoglyphic features were consistent with those of the trisomy 9p syndrome. However, the red cell galactose-1-P uridyl transferase (GALT) activity was normal despite the presence of the duplicated segment 9p13. PMID- 2984425 TI - Comparison of media with and without 'Panmede' for the isolation of Streptobacillus moniliformis from blood cultures and observations on the inhibitory effect of sodium polyanethol sulphonate. AB - Fastidious anaerobe broth and brain-heart infusion cysteine broth supplemented with 'Panmede' (a papain digest of ox liver) 2.5% supported the recovery of five Streptobacillus moniliformis strains from simulated blood cultures. Other media tested in parallel--brain heart infusion cysteine broth without 'Panmede' and Brewer's thioglycollate broth--were unreliable. Sodium polyanethol sulphonate (Liquoid) 0.05%, inhibited five isolates of S. moniliformis, including isolates from patients with Haverhill Fever. Occasionally, Liquoid 0.025% was also inhibitory and a heavy inoculum of one strain, NCTC11194, was completely inhibited by Liquoid 0.012% in simulated nutrient-broth blood cultures. These results suggest that the choice of media included in each blood-culture set is critical for the optimal isolation of S. moniliformis. Brain-heart infusion cysteine broth supplemented with 'Panmede', or commercially available fastidious anaerobe broth, without Liquoid, is recommended. PMID- 2984426 TI - Clinical comparison of anaerobic blood-culture media for detecting bacteraemia due to viridans streptococci and oral anaerobes. AB - Brain heart infusion cysteine broth, with and without Panmede (a papain digest of ox liver) and Fastidious Anaerobe Broth, with and without Liquoid, were compared by inoculating the broths with blood collected from each of 51 patients, 2 min after dental extraction. Bacteraemia caused by viridans streptococci or oral non sporing anaerobes or both was detected in 39 patients (76%). Detection of bacteraemia caused by viridans streptococci and anaerobes was more rapidly achieved by the addition of Panmede to brain heart infusion broth. Significantly more cases of bacteraemia caused by viridans streptococci were detected by use of the Panmede-containing medium than by use of Fastidious Anaerobe Broth after incubation of the broths for only 1 day. Use of brain heart infusion cysteine broth with and without Panmede, and Fastidious Anaerobe Broth permitted detection of bacteraemia caused by viridans streptococci in 26, 11 and 22 patients respectively during incubation for 2 weeks. Bacteraemia caused by anaerobes was detected by use of these three media in 24, 13 and 23 patients respectively. The addition of Liquoid to Fastidious Anaerobe Broth had no significant effect on the detection of bacteraemia caused by viridans streptococci or anaerobes. The Panmede-containing blood-culture medium should be a useful anaerobic broth in the investigation of patients with suspected endocarditis, because viridans streptococci are also rapidly detected. PMID- 2984427 TI - Abnormality of helper/suppressor T cell ratio in patients with hemophilia. AB - Lymphocyte subpopulations in patients with hemophilia were analyzed. The results were compared with those in age- and sex-matched controls. The patients had been receiving blood and blood products including Factor VIII or Factor IX for a number of years as required at the time of the onset of the disease. Heparinized peripheral blood was used in direct tests for cell marker analysis with OKT4, OKT8 and OKT11 monoclonal antibodies. There were less OKT4+ cells in the hemophiliac blood as compared with the controls, while both the groups had the same number of OKT8+ cells. As a result, the OKT4/OKT8 ratio was markedly depressed in the hemophiliac group and the conversion of the OKT4/OKT8 ratio was observed in the group of patients receiving long-term therapy with anti hemophiliac products. Alteration of lymphocyte subpopulations in patients with hemophilia in relation to antihemophiliac therapy was discussed. PMID- 2984428 TI - Radiological Seminar CCXLII: role of scintigraphy and ultrasound in evaluation of scrotal mass. PMID- 2984429 TI - Sequence determination and genetic content of the short unique region in the genome of herpes simplex virus type 1. AB - We have determined the complete DNA sequence of the short unique region in the genome of herpes simplex virus type 1, strain 17, and have interpreted it in terms of messenger RNAs and encoded proteins. The sequence contains variable regions whose length differs between DNA clones. The clones used for most of the analysis gave a short unique length of 12,979 base-pairs. We consider that this region contains 12 genes, which are expressed by mRNAs which have separate promoters, but may share 3'-termination sites, so that all but two mRNAs belong to one of four 3'-coterminal "families": 79% of the sequence is considered to be polypeptide coding. One pair of genes has an extensive out-of-frame overlap of coding sequences. The proteins encoded in the short unique region include two immediate-early species, two virion surface glycoproteins, and a DNA-binding species. Six of the genes have little or no previous characterization. From the nature of the amino acid sequences predicted for their encoded proteins, we deduce that several of these proteins may be membrane-associated. PMID- 2984430 TI - Sites of reaction of Escherichia coli DNA gyrase on pBR322 in vivo as revealed by oxolinic acid-induced plasmid linearization. AB - pBR322 DNA, linearized by lysis of an oxolinic acid-treated culture of Escherichia coli strain DK6recA- (pBR322) with sodium dodecyl sulfate, was purified, treated with DNA polymerase in the presence of the four deoxynucleoside triphosphates, and ligated to DNA linkers containing the XhoI recognition sequence. Most of the drug-resistant colonies resulting from transformation of E. coli with this material bore plasmids that appeared by restriction enzyme analysis to differ from pBR322 only by the introduction of an XhoI site. The XhoI sites in plasmids from 93 transformants were distributed unevenly around the pBR322 map. Maxam-Gilbert DNA sequence analysis of 36 of these plasmids, labeled at the 5' termini of the XhoI sites, revealed that 29 of them contained, in addition to the XhoI linker, a duplication of four base-pairs of the pBR322 sequence surrounding the linker. Therefore, oxolinic acid-induced linearization must have resulted in 5'-terminal extensions of four bases, the configuration known to result from oxolinic acid-induced DNA cleavage by DNA gyrase in vitro. The sequence data thus allowed the determination of the precise point at which linearization occurred, apparently by the abortion of a gyrase-DNA covalent intermediate that existed in vivo. When the 19 different sites of the 29 plasmids were compared, the following set of rules could be derived: (formula; see text) where N is any nucleotide, R is a purine, and Y is a pyrimidine. Cleavage occurred at the line between the eighth and ninth positions from the left. The parenthetical G and T were preferred secondarily to T and G, respectively, whereas T and G in the 13th position from the left were equally preferred. Several of these rules are similar to those proposed previously based on several in vitro gyrase cleavage sites. Some of our rules show dyad symmetry around the axis midway between the cleavage points in the two strands, while others are distinctly asymmetric. PMID- 2984431 TI - The KpnI family of long interspersed nucleotide sequences is present on discrete sizes of circular DNA in monkey (BSC-1) cells. AB - Discretely sized molecules of small circular DNAs in African green monkey kidney (BSC-1) cells contain nucleotide sequences homologous to the KpnI family of long interspersed repetitive nucleotide sequences. The size distribution of these KpnI family-containing circular DNAs differs markedly from those of BSC-1 cell circular DNAs containing either the Alu family of short interspersed nucleotide sequences or the alpha-satellite family of tandemly repeated sequences. The structures of several cloned, apparently whole, KpnI family-related circular DNAs of varying sizes were analyzed and compared with a compilation of chromosomal KpnI sequences. In general, it was found that the cloned DNAs all contained only KpnI sequences, and that the recombination events given rise to them did not involve any noticeable gain of nucleotides. PMID- 2984432 TI - Template-directed synthesis on oligodeoxycytidylate and polydeoxycytidylate templates. AB - Oligodeoxycytidylic acids and polydeoxycytidylic acid are effective templates for the polymerization of guanosine 5'-(phospho-2-methylimidazolide). They may be substituted for the corresponding ribo-oligomers without greatly changing the course of the reactions. Since oligomers of deoxynucleotides are much more easily synthesized than the ribo-oligomers, this finding, if it proves general, should greatly facilitate the study of the template properties of oligomers containing two or more bases. Oligodeoxycytidylates facilitate the synthesis of oligoguanylates up to one residue longer than the template in high yield, and oligoguanylates up to twice the length of the template in significant yield. The time-course and regiospecificity of these reactions suggest that "sliding" and "double-templating" are important factors in determining the pattern of reaction products. PMID- 2984433 TI - Analysis and optimization of recombinant DNA joining reactions. AB - The statistical segment length of duplex DNA was determined in phage T4 ligase (poly(deoxyribonucleotide): poly(deoxyribonucleotide) ligase (AMP forming), EC 6.5.1.1) buffer (50 mM-Tris . HCl (pH 7.8), 20 mM-dithiothreitol, 10 mM-MgCl2, 1 mM-ATP) at 12 degrees C to be 1030(+/- 116) A. This result was obtained by electron microscopic examination of the molecular distributions generated by T4 ligase-mediated joining of EcoRI-cleaved pBR322 DNA. This value of the statistical segment length was utilized in an extension of the Jacobson Stockmayer theory on the probability of intramolecular cyclization in order to optimize DNA joining reactions that are of great utility in recombinant DNA experiments. Five cloning systems were analyzed: circular plasmid vectors that had been linearized with one or two restriction endonucleases, circular plasmids that had been tailed with deoxyhomopolymers before joining, lambda-type cloning vectors and cosmids. The results are tabulated for convenient use in molecular cloning experiments. PMID- 2984434 TI - Dielectric studies of protein hydration and hydration-induced flexibility. AB - Dielectric measurements, as a function of hydration, are reported for collagen, cytochrome-c, elastin and lysozyme powders. The hydration dependence of the dispersion that occurs in the frequency range between 10 kHz and 10 GHz has been used to identify two classes of protein-bound water molecules (namely, rotationally hindered or unhindered), as well as the hydration level for the onset of an increasing protein flexibility. Such studies can aid an understanding of the relationship between enzyme activity and structural flexibility, and of hydration-induced changes in the structure and dynamics of protein structures. PMID- 2984435 TI - An essential replication gene, repA, of plasmid pSC101 is autoregulated. AB - Measurements of the rate of replication of a mutant pSC101 plasmid, cloned into a ColE1 vector, showed that insertions of the transposon Tn1000 into the repA gene of pSC101 abolished replication activity, but could be complemented in trans, albeit at a low level. The promoter of the repA gene was mapped by the construction of repA-lacZ gene fusions, and one of the fusions was used to demonstrate that repA protein, provided in trans, could repress expression of beta-galactosidase activity. This repression was primarily due to reduction of transcription of the repA-lacZ fusion. The sequence analysis of mutants of the repA-lacZ fusion gene which were no longer sensitive to the presence of repA protein showed that the site of action of repA was a 22 base-pair sequence, present as an inverted repeat, overlapping the repA promoter. The repA gene is thus autoregulated. PMID- 2984436 TI - Discharge instructions: providing continuity of care for ophthalmic patients. PMID- 2984437 TI - Use of repeated CO2 challenges to evaluate the pulmonary performance of guinea pigs exposed to toluene diisocyanate. AB - Guinea pigs were exposed to 1.4 ppm toluene diisocyanate (TDI) for 3 h/d on 5 consecutive days to produce chemical bronchitis. Following these exposures, the ventilatory response of the animals to challenge with 10% CO2 was greatly diminished. Response recovered during the next 40 d. In contrast, animals exposed to 0.02 ppm TDI daily for 70 d showed no change in ventilatory response to challenge with 10% CO2, nor did control animals which were not exposed to TDI. These results indicated both the absence of pulmonary toxicity following exposure to low TDI concentrations for an extended time and the effectiveness of the methodology utilizing the ventilatory response to CO2 to monitor recovery from acute pulmonary toxicity. PMID- 2984438 TI - Mixed-function oxidase system induction and propylene hepatotoxicity. AB - Propylene is hepatotoxic to male Charles River COBS Sprague-Dawley rats pretreated with polychlorinated biphenyls (PCB: Aroclor 1254). Four-hour inhalation exposure to 50,000 ppm propylene increased liver weight/body weight ratios and elevated serum enzyme activities in PCB-pretreated animals. Hepatic microsomal cytochrome P-450 content of PCB-pretreated rats dropped profoundly during propylene exposure and remained depressed for at least 24 h. In addition, PCB-pretreated, propylene-exposed rats exhibited a decrease in the specific activity of hepatic microsomal aniline hydroxylase. However, there was no change in activities of either hepatic microsomal aminopyrine demethylase or glucose-6 phosphatase. Propylene exposure of rats pretreated with beta-naphthoflavone (BNF), phenobarbital (PB), or a mixture of BNF and PB was not hepatotoxic. However, there was, in these animals, a substantial decline in hepatic microsomal cytochrome P-450 levels 24 h after the start of propylene exposure. Hence, the propylene-dependent process resulting in hepatic cytochrome P-450 destruction is qualitatively or quantitatively different from the process that causes acute hepatotoxicity. Preexposure fasting had no effect on the hepatotoxicity resulting from a 4-h exposure of PCB-pretreated rats to 50,000 ppm propylene. Administration of SKF-525A to PCB-pretreated rats immediately prior to propylene exposure completely prevented elevations in serum enzyme activities and liver weight/body weight ratios. In vitro incubation of hepatic microsomes prepared from either BNF-, PB-, or PCB-pretreated rats with an atmosphere of 20% propylene/80% air produced in NADPH-dependent decrease in cytochrome P-450 content. These results suggest that PCB pretreatment is a prerequisite for propylene hepatotoxicity in the rat. Cytochrome P-450-dependent bioactivation of propylene is associated with this hepatotoxicity, but further studies are needed to characterize the mechanism of the PCB-propylene interaction. PMID- 2984439 TI - NMR imaging and spectroscopy of experimental brain edema. AB - A canine model of experimental brain edema utilizing the classic cold lesion technique was used to identify the optimal nuclear magnetic resonance (NMR) spin echo imaging parameters needed to detect cerebral edema with maximum sensitivity. Each animal was studied with four separate spin-echo sequences, utilizing pulse intervals ranging from 0.5 to 2.0 seconds. The echo delays following each pulse interval were 28 and 56 msec. Cerebral edema was best identified with the longest pulse interval (2.0 seconds), and the second echo delay (56 msec). T1 and T2 relaxation values were obtained from in vitro spectroscopy. The relaxation times were prolonged in the edematous white matter; however, absolute T1 and T2 values proved of little use in comparing different animals. PMID- 2984440 TI - Hormonal influence on the neurogenic response of the hamster vas deferens. AB - The effect of castration on in vitro contractility of smooth muscle of the vas deferens and body of the bladder has been studied in the hamster. Castration produced supersensitivity to in vitro electrical stimulation and norepinephrine in the vas deferens, but had no effect on the body of the bladder. Castration also increased the maximum contractile response of the vas deferens to electrical stimulation, norepinephrine, ATP, acetylcholine and histamine. The changes in contractility of smooth muscle of the vas deferens developed slowly and may be explained by specific effects upon adrenergic and purinergic neurotransmission and/or non-specific effects upon smooth muscle cell membranes. PMID- 2984441 TI - Crisis in drug testing. Results of CDC blind study. AB - In response to questions about the reliability of the results of screening urine for drugs, we evaluated the performance of 13 laboratories, which serve a total of 262 methadone treatment facilities, by submitting prereferenced samples through the treatment facilities as patient samples (blind testing). Error rates for the 13 laboratories on samples containing barbiturates, amphetamines, methadone, cocaine, codeine, and morphine ranged from 11% to 94%, 19% to 100%, 0% to 33%, 0% to 100%, 0% to 100%, and 5% to 100%, respectively. Similarly, error rates on samples not containing these drugs (false-positives) ranged from 0% to 6%, 0% to 37%, 0% to 66%, 0% to 6%, 0% to 7%, and 0% to 10%, respectively. These blind tests indicate that greater care is taken with known evaluation samples than with routine samples, laboratories are often unable to detect drugs at concentrations called for by their contracts, and the observed underreporting of drugs may threaten the treatment process. Drug treatment facilities should monitor the performance of their contract laboratories with quality-control samples, preferably through blind testing. PMID- 2984442 TI - HTLV-I antibodies in childhood leukemia. PMID- 2984444 TI - [A case of signet ring cell carcinoma of the urinary bladder]. AB - A rare vesical carcinoma, arising in a 47-year-old male, is described. The tumor developed from the left ureteral orifice and posterior urethra, showing a nonpapillary appearance with a wide stalk. By light microscopy, the carcinoma was found to be mainly composed of signet ring cells and associated with small foci of transitional cell carcinoma. The previously reported cases of signet ring cell carcinoma of the urinary bladder were reviewed with a special reference to the manifold potentiality of the urothelium. PMID- 2984443 TI - [Current perspectives in the management of small cell lung cancer]. AB - Considerable progress has been made within the last decade in the management of small cell lung cancer (SCLC) resulting in prolongation of median survival by 4 to 5 times (about 14 and 9 months in limited and extensive disease, respectively), improved quality of life, and an increase of cure rates in 15 approximately 20% of the patients with limited disease. In this review, we are focusing on the details in update treatment schedules of combination chemotherapy against SCLC, prognostic features and staging of SCLC, the role of non-cross resistant alternating chemotherapy, radiotherapy to the primary site including hilar and mediastinal lymph nodes, prophylactic cranial irradiation and surgery in the treatment of SCLC, as well as the complications of treatment. PMID- 2984445 TI - An unusual case of gastric carcinoma initially presenting bone metastasis and later transverse myelopathy. AB - A 44-year-old man with Borrmann type III gastric carcinoma initially presenting bone metastasis and later transverse myelopathy is reported. Chest and skull roentgenograms showed multiple punched-out lesions. A bone scintigram showed multiple abnormal uptake. Histological examinations of biopsy and autopsy materials revealed signet-ring cell carcinoma of the stomach. Disseminated carcinomatosis of the bone marrow was ruled out histologically and neither microangiopathic hemolytic anemia nor disseminated intravascular coagulopathy was observed. Because this patient with bone metastasis did not have metastasis to the liver and lungs we propose that a nonportal route through the vertebral venous plexus might be an alternative to the portal route of bone metastasis from gastric carcinoma. During the last 20 years, only 10 such cases including this one have been reported in Japan. Their prognosis was quite poor and in most of them the malignancy was either undifferentiated adenocarcinoma or signet-ring cell carcinoma. If a bone metastasis is found as the initial sign, physicians should keep in mind that the primary lesion might be in the stomach, or in the thyroid, kidney, lung or prostate as another point of search. PMID- 2984446 TI - Malignant fibrous histiocytoma in a fox. PMID- 2984447 TI - Ascitic disease in nude mice infected with mouse hepatitis virus. PMID- 2984448 TI - Adenylate kinase from rat liver: molecular properties and structural comparison with yeast enzyme. PMID- 2984450 TI - [Reaction of cyclic plasma nucleotides of healthy subjects and essential hypertension patients to insulin stimulation]. AB - A total of 112 patients with borderline arterial hypertension and essential hypertension of the labile and stable forms were examined. The plasma levels of cyclic AMP and cyclic GMP as well as the renin activity of the blood were determined in all those studied both at rest and following insulin administration. The findings obtained point to a certain role of cyclic nucleotides in the development and stabilization of arterial hypertension. PMID- 2984449 TI - [Concentration of adrenocorticotrophic hormone and aldosterone secretion in essential hypertension and hyperaldosteronism]. AB - The authors studied the effect of adrenocorticotropic hormone (ACTH), potassium and plasma renin activity on blood aldosterone in normal subjects as well as in patients with essential hypertension (of a labile and stable course) and hyperaldosteronism (primary and idiopathic). It was demonstrated that in normal subjects and patients with labile essential hypertension, the secretion of aldosterone was simultaneously stimulated by the renin-angiotensin system (RAS) and the hypothalamus-adenopituitary. The RAS dominated in normal conditions whereas in labile hypertension the hypothalamus-adenopituitary system was predominant. In stable hypertension, the RAS and hypothalamus-pituitary influenced aldosterone secretion in an equal degree. Hyperaldosteronism was associated with the most pronounced deviations in the relationship between stimulants and aldosterone. In addition to decreased plasma levels of renin activity and potassium, the corticotropic activity of the hypothalamus adenopituitary was increased during the first 10 years of the disease, while later on the function of this system became inhibited. The highest ACTH levels were recorded in idiopathic hyperaldosteronism. PMID- 2984451 TI - Effect of cyclic AMP on acidification in the isolated turtle bladder. AB - Cyclic AMP (10 mM) has been demonstrated to inhibit hydrogen ion secretion in the isolated turtle bladder. These experiments were designed to study the effect of cyclic AMP on hydrogen ion secretion in the isolated turtle bladder using both the pH stat and the reverse short circuit current techniques. Sodium transport was measured as the short circuit current. Studies were carried out at 0% CO2 and 1% CO2 at pH 7.4. Cyclic AMP (10 mM) was added to either the serosal or mucosal solutions, and hydrogen ion secretion was measured from 0 to 120 min. In the presence or absence of carbon dioxide, cyclic AMP and dibutyryl cyclic AMP had no effect on hydrogen ion secretion in fasted turtles. The addition of theophylline (10 mM) to the serosal solution, with or without cyclic AMP had no effect on proton secretion. Sodium transport was unchanged from control following serosal or mucosal addition of 10 mM cyclic AMP in the presence or absence of carbon dioxide. In chronically bicarbonate-loaded turtles proton secretion was the same as control fasted turtles. In these animals, however, serosal administration of 10 mM cyclic AMP significantly stimulated bicarbonate secretion. Stimulation of bicarbonate secretion occurred in the presence of a 20 mM bicarbonate gradient. When there was no bicarbonate gradient, cyclic AMP was without effect; cyclic AMP had no effect on bicarbonate permeability when measured in the presence of acetazolamide. These results indicate that cyclic AMP has no effect on hydrogen ion secretion or sodium transport in the isolated turtle bladder when studied at two different rates of acidification (0 and 1% CO2). Cyclic AMP appears to stimulate active bicarbonate secretion. PMID- 2984452 TI - Effect of urinary pH and diatrizoate on Bence Jones protein nephrotoxicity in the rat. AB - Both low urinary pH and radiocontrast agents may intensify myeloma nephrotoxicity. To study the effects of these factors, we determined inulin clearances (CIn) before and after infusions of human Bence Jones protein (BJP) in male Sprague-Dawley rats in a dose previously shown to be nephrotoxic. Rats that drank 0.15 M NaHCO3 for 48 hr before study had no change in CIn (+3 +/- 20%) after BJP unlike those that drank 0.15 M NH4Cl (-33 +/- 14%, P less than 0.05); urinary pH differed (7.6 +/- 0.1 vs. 6.2 +/- 0.1, P less than 0.05), but urinary flow rates did not. The acidifying regimen was used in all subsequent groups. Infusion of diatrizoate (DTZ) after BJP produced a further decrease in CIn (-85 +/- 8%, P less than 0.05). In contrast, infusion of albumin, which raised plasma protein concentration to that seen in BJP-infused rats, did not change CIn (+39 +/- 17%). Infusion of beta-lactoglobulin also led to a greater decrease in CIn after DTZ (-35 +/- 9 vs. -67 +/- 8%, P less than 0.05), but myoglobin did not ( 58 +/- 7 vs. -54 +/- 12%). Urinary pH and flow rate did not differ between any DTZ-infused group and its appropriate control. These data suggest that aciduria independent of urinary flow rate increases the nephrotoxicity of BJP. In this setting, DTZ further intensifies the nephrotoxicity of BJP as well as some but not all filterable proteins. PMID- 2984453 TI - Effects of bilirubin on transepithelial transport of sodium, water, and urea. AB - Urinary concentrating defects and renal salt wasting have been described in the hyperbilirubinemic Gunn strain strain of rat. Homozygous animals demonstrate significant reductions in renal medullary urea and sodium ion concentrations. These observations are consistent with possible bilirubin associated disorders in the transepithelial transport of water and solute. To test this hypothesis, measurements of active sodium transport and passive water and urea fluxes were made in hemibladders isolated from the Dominican toad, Bufo marinus. Tissues were exposed to amphibian bicarbonate Ringer's solution containing 0.1 mM bilirubin with 0.05% bovine serum albumin (BSA) or BSA alone. Vasopressin-stimulated sodium transport, as reflected by short circuit current (SCC), was inhibited by 18 +/- 6% in the presence of bilirubin (N = 10; P less than 0.02). Cyclic AMP (p-Cl phenylthio cAMP 10(-5) M) stimulated SCC was inhibited to a similar degree in the presence of bilirubin. The inhibition was noted only when bilirubin was in the serosal bath, and it could be abolished with BSA 0.5%. Bilirubin had no effect on the increase in SCC induced by higher concentrations of cyclic AMP (10(-4) M), aldosterone, or amphotericin B. Furthermore, bilirubin had no effect on the hydro osmotic response to vasopressin and vasopressin-induced changes in urea permeability. These findings show that short-term exposure to bilirubin exerts a tissue-specific effect on the vasopressin-stimulated active transport of sodium but has no effect on the vasopressin-induced fluxes of water and urea. PMID- 2984454 TI - Detection of coronavirus-like particles in homosexual men with acquired immunodeficiency and related lymphadenopathy syndrome. AB - Coronavirus-like particles were identified by electronmicroscopy in the feces of homosexual men. The particles banded at a density of 1.21 g/ml after cesium chloride density gradient centrifugation. To determine whether the presence of this virus might be related to clinical symptoms, several patient groups were studied prospectively. In 8 of 16 (50%) homosexual males with acquired immunodeficiency syndrome (AIDS) or unexplained lymphadenopathy syndrome (LAS), coronavirus particles were found. In contrast, such particles were found in none of 18 heterosexual controls and in only 3 of 20 homosexual males without AIDS or LAS. Thus, coronavirus excretion correlated significantly (2 alpha less than 0.01) with the clinical diagnosis of AIDS or with syndromes belonging to the AIDS related complex. In addition, such particles identified in the serum of one patient with LAS and diarrhea suggest invasion and systemic spread of the agent and underline that this virus behaves differently from "common cold" human coronaviruses. PMID- 2984455 TI - Physical activity decreases the number of beta-adrenergic receptors on human lymphocytes. AB - On intact human lymphocytes a specific binding site (BS) for 125I-Cyanopindolol (125I-CYP), a derivative of the beta-blocking drug pindolol, was characterized. Inhibition of binding for catecholamines in the following order of potency: l isoprenaline greater than l-adrenaline greater than l-noradrenaline proves the BS as a beta 2-receptor subtype. In 77 healthy persons (36 females, 41 males) the number of BS amounted to 2,639 +/- 125 BS/cell without any significant correlation to age (17-86 years) or sex. The dissociation constant (KD) indicating the affinity of iodocyanopindolol to the BS on intact lymphocytes was KD = 1.9 +/- 1.1 X 10(-10) M. A change of the number of beta-adrenergic receptors on intact human lymphocytes has been measured previously in asthmatics and in the myocardium of patients with congestive heart failure. We investigated a possible change in the beta-receptors on lymphocytes by physical and mental activity ("stress") in physicians going about their daily routine work. Persons left alone in a hospital room reading or sleeping were defined as "inactive controls". The number of BS on intact lymphocytes was significantly higher in inactive persons at 8 a.m. (2,230 +/- 482 BS) compared to active persons (1,743 +/- 285 BS; P less than 0.05) and at 1 p.m. (2,394 +/- 253 BS vs 1,733 +/- 556; P less than 0.05) but not different at 6 p.m. (1,634 +/- 578 BS vs 1,768 +/- 588 BS; P less than 0.1). The KD remained unchanged under all conditions. The serum noradrenaline and adrenaline levels were also measured during the day.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984457 TI - [Study visit to a cancer after-care hospital: Sonneberg Hospital Bad Sooden Allendorf]. PMID- 2984458 TI - Kinetics of ectromelia virus (mousepox) transmission and clinical response in C57BL/6j, BALB/cByj and AKR/J inbred mice. AB - Research was undertaken to answer basic questions on susceptibility, clinical response and transmission of ectromelia virus in selected strains of inbred mice. C57BL/6J and AKR/J were found to be markedly more resistant to a virulent strain of ectromelia virus (isolated during the 1979-80 outbreak at the National Institutes of Health) than C57LJ, BALB/cByJ, DBA/2J, A.By/SNJ and C3H/HeJ when infected by footpad inoculation. In C57BL/6J and AKR/J the LD50 was about 7 logs higher than the ID50. With one exception, C57LJ, the LD50 and ID50 titers in the other strains were about equal. In C57LJ the LD50 titer was intermediate. Following intragastric inoculation, virus was isolated from feces of C57BL/6J mice for as long as 46 days and up to 29 days from BALB/cByJ mice. Transmission to cage mates from intragastrically infected C57BL/6J and BALB/cByJ occurred up to 36 and 30 days respectively after infection. Virus was isolated from the spleen in 2 of 5 BALB/cByJ mice and 1 of 7 C57BL/6J mice tested 95 days after gastric inoculation. Following footpad inoculation, BALB/cByJ mice consistently transmitted virus to cage mates before death at 10-12 days. C57BL/6J mice transmitted between days 8 and 17, but not beyond. Virus was maintained in C57BL/6J mice by exposure to infected cage mates for seven passages, which was the most attempted. Clinical signs in infected C57BL/6J mice were usually subtle or inapparent. PMID- 2984456 TI - [Importance of fibroblast chemotaxis in wound healing and tumor cell evasion]. AB - Fibroblast are responsible for the synthesis of the structural proteins of the connective tissue. A further property of these cells, their migratory ability, could be analyzed in the last years. A special form of migration is chemotaxis, which can be quantitatively measured in a modified Boyden chamber in-vitro. Using this method chemoattractive substances could be characterized, which are able to stimulate fibroblasts and tumorcells to chemotactic migration. Furthermore it could be proved, that benign and transformed cell lines react in a different manner towards these chemoattractive substances. The in-vitro results allow some hypotheses about both fibroblast migration in wound healing or chronic inflammation, and the mechanisms of tumor cell evasion in the tumor surrounding tissue or the metastasizing process in other organs. PMID- 2984459 TI - Diagnostic exercise: subcutaneous nodules in rhesus monkeys. PMID- 2984460 TI - Disseminated cytomegalovirus disease in the guinea pig. AB - Two guinea pigs (Cavia porcellus), which were housed conventionally in separate animal facilities and had not been experimentally manipulated, were found to have evidence of disseminated cytomegalovirus disease at necropsy. Microscopic examination revealed multiple focal areas of necrosis in numerous organs. The spleen, liver, kidney and lungs were affected particularly. These lesions contained numerous cells with large intranuclear and cytoplasmic inclusion bodies resembling cytomegalovirus inclusions. Characteristic cytomegalovirus virions were observed by electron microscopy in both cases. Antigens of guinea pig cytomegalovirus were detected in paraffin sections of lesions from both cases with an immunoperoxidase technique. Although subclinical infection is common, this is the only contemporary report of disseminated disease due to cytomegalovirus in guinea pigs. PMID- 2984461 TI - Some characteristics of hyperoxia-adapted HeLa cells. A tissue culture model for cellular oxygen tolerance. AB - By culturing HeLa cells at stepwise increased oxygen tensions over a prolonged period of time (approximately 21 months) we selected a substrain capable of growing under 80% O2/19% N2/1% CO2, an oxygen level that is lethal to normal HeLa cells, adapted to 20% O2/79% N2/1% CO2. The 80% O2-adapted cells exhibited the following characteristics. At the ultrastructural level an abnormal mitochondrial morphology was observed: compared to normal cells, mitochondria of the hyperoxia adapted cells exhibited a 3-fold larger mean profile area in sections and were slightly decreased in number; the relative mitochondrial volume was increased 2 fold, whereas the size of both cell types was the same. Mitochondrial matrix appeared less dense in the hyperoxia-adapted cells; no structural damage was detected. Compared to the 20% O2-adapted cells O2 consumption per cell was approximately 40% decreased in the 80% O2-adapted cells. Under hyperoxic conditions 20% O2-adapted and 80% O2-adapted cells exhibited very similar cyanide resistant respiration rates (0.16 +/- 0.04 and 0.15 +/- 0.02 fmoles/cell/minute, respectively), suggesting that the increased O2 tolerance of the 80% O2-adapted cells was not due to a decreased cellular production of activated oxygen species at hyperoxia. Cellular levels of the enzymes directly involved in protection against activated oxygen species, i.e., superoxide dismutases, catalase, and glutathione peroxidase, were normal or slightly below normal in the 80% O2 adapted cells, implying that these enzymes were of no significance for the increased O2 tolerance. In addition, the specific activity of glucose-6-phosphate dehydrogenase, a key enzyme for cellular production of NADPH, was not related to the degree of O2 tolerance. Our results suggest that the increased O2 tolerance of the 80% O2-adapted cells is neither based on cellular properties controlling the formation or removal of intracellular activated oxygen species nor on the cellular capacity to repair or replace damaged cellular components. We speculate that the increased O2 tolerance is largely due to a genetically determined increased resistance of oxygen-sensitive cellular targets. PMID- 2984462 TI - Neutral metalloendopeptidase in human male genital tract. Comparison to angiotensin I-converting enzyme. AB - High concentrations of neutral metalloendopeptidase (NEP) (enkephalinase) were found in human male genital tract immunohistochemically and by enzyme activity assays, and its distribution was compared with that of angiotensin-converting enzyme (ACE) (kininase II). Whereas the two enzymes colocalize on the luminal aspect of proximal tubular epithelium and are not found elsewhere in the nephron, their distribution in the male genitalia is different. Seminal fluid is rich in NEP and ACE, but after ultracentrifugation ACE remains soluble while NEP sediments. NEP activity is low in testicular homogenate but high in the particulate fraction of epididymides and prostates. ACE, on the other hand, is active in the particulate fraction of testes and in the soluble fraction of epididymides and prostates. Prostatic NEP had a slightly higher molecular weight than the renal NEP, which was reduced by neuraminidase in electroblotting. Testicular and seminal plasma ACE also had a slightly higher molecular weight than the purified renal enzyme (150,000), probably caused by removal of an "anchor" peptide during purification. In the prostate, NEP was found by three different immunohistochemical techniques in luminal epithelial cells and in lumina. The function of NEP in the genital tract may be related to sperm maturation and proacrosin activation. PMID- 2984463 TI - Detection of recent cannabis use by saliva delta 9-THC radioimmunoassay. AB - A non-invasive saliva sample delta 9-THC radioimmunoassay has been applied to 352 samples from 25 male and 10 female marijuana users after administration of one half to two standard cigarettes (27 mg delta 9-THC/cigarette) and 72 control negative subjects who ingested a large variety of foods, condiments, or medications in an attempt to demonstrate interferences. The shortest duration of a positive was 2 hrs and the longest was 5 hrs after administration of the cannabis. No positives occurred in control subjects. PMID- 2984464 TI - A semi-automated extraction and spotting system for drug analysis by TLC. I. Procedure for analysis of the major metabolite of delta 9-tetrahydrocannabinol in urine. AB - A procedure is described for the detection and quantitative analysis of the major metabolite of tetrahydrocannabinol in urine. Following alkaline hydrolysis of the conjugate, the metabolite is extracted, eluted, and spotted on a high performance silica gel thin layer chromatographic plate using a semi-automatic apparatus. The method has an average recovery of 95.9% and an average precision of 6.3%. A study of 32 drugs and metabolites showed no interferences. The method has a visual detection limit at 100% reliability of 2 ng/ml using 2 mL of spiked urine. It was 100% accurate at detecting authentic samples which were positive by the Enzyme Multiplied Immunoassay Technique. There was a good correlation between urine samples analyzed by gas chromatography/mass spectroscopy and the TLC procedure. The method is suitable for the simultaneous analysis of 60 samples. PMID- 2984465 TI - Suppression of murine immunologic functions by fresh and cultured human tumor extracts. AB - The role of human tumor-derived immunoregulatory factors (IRF) in the suppression of murine in vitro cell-mediated immune systems was investigated. IRF was extracted from a fresh human colon carcinoma and a liposarcoma using 3 M KCl. These extracts have previously been shown to suppress in vitro human immune responses. Both IRF extracts inhibited PHA-stimulated murine splenocyte [3H]Tdr uptake in a dose-dependent manner while extracts of normal tissue were not inhibitory. To further investigate in vitro immunosuppression a (C57BL/6 X A/J) F1 anti-B10. BR mixed lymphocyte reaction (MLR) was developed. Optimal [3H]Tdr incorporation was on Day 4 with 1 X 10(5) responders and 2 X 10(5) irradiated stimulators. Addition of IRF caused a 56% inhibition of this response but did not alter the kinetics of the MLR response. Induction of cell-mediated cytotoxicity (C57BL/6 X A/J F1 vs B10.D2) was significantly inhibited by addition of IRF during in vitro sensitization. Release of 51Cr from P-815 targets was decreased to spontaneous release levels at an effector:target (E:T) ratio of 20:1 when IRF was present during sensitization. At this E:T ratio, cells sensitized in the presence of a normal muscle 3 M KCl extract or medium caused 71 and 60% 51Cr release, respectively. IRF activity could also reproducibly be extracted from two small cell lung carcinoma tissue culture lines grown under a variety of culture conditions or passaged in nu/nu mice. The biochemical characteristics of the factor inhibiting human and murine lymphoid cell proliferation were identical. Thus, this system provides a convenient model for assessing the activity of human tumor-derived immunoregulatory factors. PMID- 2984466 TI - Bilateral Wilms' tumor: 18-year follow-up of patient treated only with surgery. PMID- 2984467 TI - Inhibitory or stimulatory effect of human genes on the expression of adrenal function in human Leydig X Y1 cell hybrids. AB - In order to investigate the expression and the regulation of steroidogenesis, human Leydig cells were fused with a functional mouse adrenal cell line (Y1). Six independent hybrid clones were analysed for hormone receptors and for cAMP and steroid response to ACTH, hCG, 8Br-cAMP or forskolin. All hybrids had lost hCG receptors and their ability to produce testosterone. With respect to the response of adenylate cyclase to ACTH and/or forskolin, hybrids could be classed into two groups. In the first group, the pattern of response was qualitatively similar to Y1 parental cells; The second group was far less responsive to ACTH than are Y1 cells, and when added together, forskolin and ACTH only had an additive effect. All hybrids responded to ACTH and 8Br-cAMP with an increased production of pregnenolone (P5). The amounts of P5 produced both under basal conditions and following 8Br-cAMP stimulation were significantly higher in three hybrids when compared to Y1 cells. However, the ability of two of these three hybrids to produce 20 alpha-dihydroprogesterone (20 alpha OHP4) was very low. The metabolism of [14C]P5 revealed that in one of these hybrids, there was a loss of 3 beta hydroxysteroid dehydrogenase/isomerase whereas in the other case, there was a low 20 alpha-hydroxylase activity. The inhibition of cell growth by ACTH was related to the ability of the hormone to stimulate cAMP. Conversely, the inhibitory growth effects of 8Br-cAMP were not always inversely correlated with the ability of this nucleotide to stimulate P5 production. Since hybrids contained two mouse genomes and retained variable human chromosomes, these results suggest that extinction or enhancement of murine genes coding for some of the enzymes involved in steroidogenic response to ACTH was due to the regulation by human genes. PMID- 2984469 TI - The role of ACTH in determining the metabolic pathways of deoxycorticosterone by newborn rat adrenal cells in primary culture. AB - The metabolism of deoxycorticosterone (DOC) by newborn rat adrenal cells in primary culture at various times after culture, with and without ACTH, was studied. After 5 days in culture before addition of ACTH, the main products of the metabolism of DOC were corticosterone and 18-hydroxy-11-deoxycorticosterone in a 2:1 ratio. Smaller amounts of 20 alpha-dihydrocorticosterone and 18 hydroxycorticosterone were also found. No reduced metabolites of DOC were detected. Without ACTH the conversion of DOC to corticosterone and 18-hydroxyDOC declined rapidly. After 13 days in culture, this conversion accounted for only half the metabolites. The reductive metabolism of DOC which yields products reduced at 20 alpha and/or 3 alpha/beta and 5 alpha accounted for the other half. When ACTH (22 mU/ml) was added to the culture daily for several weeks, the primary metabolism of DOC remained that of 11 beta- and 18-hydroxylation yielding corticosterone and 18-hydroxyDOC. A minor reductive metabolism was found. Both cultures produced 6 beta-hydroxyDOC. These results demonstrate that ACTH is needed to maintain the efficiency of the 11 beta/18-hydroxylating system. They also show that ACTH controls the type of metabolism predominant in the rat adrenal cell and may be responsible for the balance between the biosynthesis of glucocorticoids and their reductive catabolism in the fasciculata zone of the adrenal gland. PMID- 2984468 TI - Responses of rat adrenal glomerulosa and inner zone cells to synthetic ACTH analogs and proopiomelanocortin-derived peptides. AB - A comparison of the responsiveness of isolated rat adrenal decapsular and glomerulosa cells to corticotrophin 1-39 (ACTH 1-39), synthetic ACTH analogs (characterized by a shorter amino acid chain length, the substitution of certain amino acids in the natural sequence by other amino acid residues, the replacement of the C-terminal carboxyl group by an amide), and proopiomelanocortin-derived peptides was performed by measuring corticosterone and aldosterone production, respectively. The potencies of the synthetic ACTH analogs correlated closely with the length of the peptides, similarly in both zones. No activity was observed with the proopiomelanocortin-derived peptides in either zone, with the exception of beta-LPH and alpha-MSH. PMID- 2984470 TI - Isolated adrenocortical cells of the domestic fowl (Gallus domesticus): steroidogenic and ultrastructural properties. AB - Isolated adrenocortical cells from White Leghorn chickens (Gallus domesticus) were compared to those from rats (Rattus norvegicus). Cells were prepared from collagenase-dispersed adrenal glands of sexually mature male animals. Corticosterone was measured by radioimmunoassay after incubation for 2 h with steroidogenic agents. Of the four ACTH analogues used, three were 6-17 times more potent with rat cells than with fowl cells (potencies were indicated by half maximal steroidogenic concentrations). However, 9-tryptophan (O nitrophenylsulfenyl) ACTH was 8 times more potent with fowl cells than with rat cells, thus suggesting that ACTH receptor differences exist between the two cell types. In addition, cAMP analogues were 10 times more potent with rat cells than with fowl cells suggesting that fowl corticosteroidogenesis is less dependent on cAMP than is rat corticosteroidogenesis. At equal cell concentrations, rat cells secreted 20-40 times more corticosterone than did chicken cells when they were maximally stimulated. Although rat cells converted 8 times more pregnenolone to corticosterone than did fowl cells, the half-maximal steroidogenic concentration for pregnenolone-supported corticosterone synthesis was the same for both cell types (about 5 microM). This suggests that fowl cells have lower steroidogenic enzyme content rather than lower steroidogenic enzyme activity. An unusual feature seen in the isolated fowl adrenocortical cells was an abundance of intracellular filaments. PMID- 2984472 TI - Malignant fibrous histiocytoma of solitary jejunal diverticulum. AB - A rare case of malignant fibrous histiocytoma arising from a solitary jejunal diverticulum leading to its perforation is described in a 37-year-old male. The patient presented with peritonitis and on exploration revealed perforation of a jejunal diverticulum containing a soft, friable mass. Biopsy from the mass revealed a malignant fibrous histiocytoma. This appears to be the first description in literature of a malignant fibrous histiocytoma with that particular localization. PMID- 2984471 TI - Specific antisera for the radioimmunoassay of estradiol-3-sulfate. AB - Antisera were prepared against two types of estradiol-3-sulfate-bovine serum albumin (BSA) conjugates. The haptens were coupled to BSA through the C-6 position in the steroid molecule by the glutaraldehyde (A) or the carbodiimide method (B). In comparison the antiserum produced by method A had a high affinity for estradiol-3-sulfate (Ka = 5.64 X 10(8) M-1); that produced by method B had an even higher affinity (Ka = 2.62 X 10(9) M-1). Furthermore the latter had no significant cross-reaction with other estrogen sulfates (less than 3.83%), and no cross-reaction with other steroids (less than 0.03%). The former revealed a little cross-reactivity with some of related steroids. PMID- 2984473 TI - Paget's disease of the anal mucosa in association with carcinoma demonstrating mucoepidermoid features. AB - A case of Paget's disease of the anal mucosa with an underlying carcinoma demonstrating mucoepidermoid features is reported. The histopathologic characteristics of both extramammary Paget's disease and mucoepidermoid carcinoma are noted. Special histochemical staining procedures were consistent with the diagnosis. The actual histogenesis of extramammary Paget's disease and mucoepidermoid carcinomas are controversial topics that are briefly discussed in this paper. PMID- 2984474 TI - Giant intraductal papilloma of the breast: a case report. AB - A case of an unusually large solitary intraductal papilloma of the breast is reported. This benign neoplasm presented as a 15-cm breast mass, enlarging over a 2-year period. The case illustrates that benign intraductal papilloma may rarely present as a large breast mass and therefore should be considered in the differential diagnosis of very large breast lesions. PMID- 2984475 TI - Our experience with resection of primary and secondary liver tumours. AB - The authors retrospectively analyse the results achieved in recent years by surgical treatment of 103 malignant primary or secondary liver cancers: (Three patients have been operated on twice at different times for recurrence.) 23 hepatic resections were performed for benign lesions. The operative mortality is very low, the incidence of postoperative complications is minimal, and the survival up to 8 years is rather satisfactory. In all cases we used Ton That Tung's surgical technique, that is, "finger fracture with intraparenchymal ligature of the vasculobiliary pedicles." PMID- 2984476 TI - The rat urinary bladder. A novel preparation for the investigation of central opioid activity in vivo. AB - A novel method, which utilized spontaneous, volume-induced contractions of the urinary bladder in the anesthetized rat, was developed to assess the central activity of substances with opioid properties. It was discovered that intracerebroventricular (i.c.v.) injections made directly into the lateral ventricle or intrathecal (i.t.) injections made into the spinal subarachnoid space inhibited bladder contractions in a dose-dependent manner. The methods for spinal and supraspinal drug administrations are described as is the estimation of the spread of drug as determined by i.c.v. and i.t. dye injections. The pharmacology of the i.c.v. and i.t. effects of opioids was determined by use of selective opioid agonists (mu-agonists: morphine, DAGO; delta-agonists: DADLE, DPDPE; kappa-agonist: U-50, 488H) and antagonists (mu-antagonist, naloxone; delta antagonist, ICI 174,864). It was concluded that the i.c.v. and i.t. effects were mediated by mu- and delta-opioid receptors and not by kappa-receptors. A number of other agents such as pentobarbital, haloperidol, and desipramine also inhibited bladder activity following i.c.v. and i.t. administration, but their actions were insensitive to naloxone. This in vivo model was considered useful in the detection and evaluation of the central actions of substances with opioid properties. PMID- 2984477 TI - Cross-species gene transfer; implications for a new theory of evolution. AB - It has been established that genes can be transferred and expressed among procaryotes of different species. I am hypothesizing--and there is mounting evidence for this conclusion--that genes are transferred and expressed among all species, and that such exchange is facilitated by, and can help account for, the existence of the biological unities, from the uniform genetic code to the cross species similarity of the stages of embryological development. If this idea is correct, the uniformity of the genetic code would allow organisms to decipher and use genes transposed from chromosomes of foreign species, and the shared sequence of embryological development within each phylum would allow the organism to integrate these genes, particularly when the genes affect complex morphological traits. The cross-species gene transfer model could help explain many observations which have puzzled evolutionists, such as rapid bursts in evolution and the widespread occurrence of parallelism in the fossil record. PMID- 2984478 TI - Assessment of capacitor electrodes for intracortical neural stimulation. AB - Capacitor electrodes offer the potential for the safest method of stimulation of neural tissue because they operate without any faradaic process occurring at the electrode-electrolyte interface. Their use eliminates problems associated with metal dissolution or water electrolysis which may occur with electrodes of noble metals. This paper reviews recent work aimed at increasing the charge storage density of capacitor electrodes to allow their application with the small areas of 10(-4) mm2 required for intracortical stimulation of single neurons. Increased charge storage with electrodes using anodic films such as TiO2 and Ta2O5 has been obtained by increasing the real surface area of microelectrodes. Experiments have also been done with BaTiO3 films which have a much higher dielectric constant than the anodic film dielectrics. State-of-the-art electrodes made with these materials, however, have a charge storage density which at best is comparable to that obtained with Pt and is considerably lower than electrochemically safe charge densities that have been reported for activated Ir. It is concluded that for very small intracortical electrodes, capacitor electrodes will not be competitive with electrodes which operate using surface localized faradaic reactions. PMID- 2984479 TI - High-current and high-voltage stimulators. PMID- 2984480 TI - Signal averaging by microcomputer using a program written in a high-level language. AB - A program, written in the language Pascal, is described for signal averaging with a microcomputer. The program instructs the computer to acquire data from two channels on-line. For up to 3 channels of extracellular or intracellular recording analysed simultaneously, it was found not to be necessary to employ machine code or an assembler to provide adequate temporal resolution. PMID- 2984481 TI - A method for recording agonist-induced depolarizations in small autonomic ganglia. AB - A convenient extracellular method is reported for recording from autonomic ganglia as small as the chick ciliary ganglion. This technique allows recording of both the compound action potentials arising from synaptic transmission through the ganglion and ganglion depolarizations due to such agents as carbachol or potassium. A perfusion system is also described that allows rapid introduction and removal of pharmacological agents. The principal advantages of the technique are two-fold: (1) the method is simple and should work for any small ganglion as long as recordings can be made from the postganglionic nerve using a suction electrode; (2) the rapid introduction and removal of agonists allows frequent measurements of ganglionic depolarization with little receptor desensitization. PMID- 2984482 TI - The differential inhibitory effect of lymphotoxin and immune interferon on normal and malignant lymphoid cells. AB - Supernatants containing lymphotoxin (LT) and immune interferon (IFN-gamma) or IFN gamma alone were produced by antigen-stimulated murine T cell clones. These lymphokine preparations as well as cloned recombinant murine IFN-gamma (Genentech) were tested for antiproliferative activity on a variety of murine T, B, macrophage, and mastocytoma tumors and on T cell clones and LPS-stimulated B cells. The growth of every cell line tested was susceptible to the LT-containing supernatants. Cloned recombinant murine IFN-gamma inhibited the growth of A9 fibroblasts but not L929 cells. Neither the cloned murine IFN-gamma nor that produced by a T cell clone had any appreciable effect on the lymphoid cells except for one B cell lymphoma, A20. The sensitivity of nontransformed T and B cells to LT indicates that this lymphokine may play an immunoregulatory role. Indeed, the T cells that produce LT are sensitive to its cytotoxic activity and, therefore, regulate themselves. The differential inhibitory effects of LT and IFN gamma on lymphoid target cells allow us to distinguish between preparations that contain IFN-gamma alone and those that contain LT and IFN-gamma. The susceptibility of lymphoid tumors to the antiproliferative activity of the LT containing preparations indicates that LT either alone or together with IFN-gamma may be useful in tumor therapy. PMID- 2984484 TI - Can plasma human pancreatic polypeptide be used to detect diseases of the exocrine pancreas? AB - Plasma concentrations of human pancreatic polypeptide (HPP) parallel exocrine pancreatic secretion in response to stimulation with cholecystokinin. We determined prospectively the relationships among fasting HPP level, integrated HPP response to infusion of cholecystokinin, and output of trypsin and also the sensitivity, specificity, and predictive values of the fasting HPP level in the diagnosis of exocrine pancreatic disease. Our study group consisted of 19 patients with acute pancreatitis, 17 with chronic pancreatitis, and 25 with ductal adenocarcinoma of the pancreas and 27 control subjects. In the control patients and those with chronic pancreatitis, significant correlations were detected between HPP level and output of trypsin (P less than 0.001) in response to infusion of cholecystokinin and between fasting HPP and integrated HPP levels (P less than 0.004); no correlation was detected between HPP level and steatorrhea. The sensitivity, specificity, and negative and positive predictive values of the fasting HPP level for detection of either chronic pancreatitis or pancreatic cancer were similar and approximated 0.88, 0.67, 0.88, and 0.66, respectively. The HPP concentration had no value in detecting acute pancreatitis. Because the fasting HPP level has a high degree of negative predictability and is simpler to measure than the integrated HPP level or the output of trypsin, it may be a useful test in patients suspected of having either chronic pancreatitis or pancreatic cancer. A fasting HPP level of 125 pg/ml or greater could be used to exclude chronic pancreatitis or pancreatic cancer, but the finding of a value of less than 125 pg/ml necessitates use of other diagnostic tests for reliable determination of the presence of these diseases. PMID- 2984485 TI - Construction of a functional surgical stent for injectable hydroxylapatite. PMID- 2984483 TI - Surgical options for patients with deep-seated brain tumors: computer-assisted stereotactic biopsy. AB - The histologic nature of deep-seated intracranial lesions can be determined by using a computer-assisted stereotactic biopsy technique. The procedures are performed with use of local anesthesia. A data base consisting of stereotactic computed tomographic scans and stereotactic cerebral angiography is acquired. Target coordinates and trajectory approach angles are calculated by using a computer system in the operating room. Since July 1984, 36 patients with a variety of pathologic lesions in various intracranial sites have undergone this procedure at our institution. Of the 36 patients thought to have neoplastic lesions preoperatively, 6 were found to have nonneoplastic lesions, information that was of importance in the therapeutic management of these patients. Of the 30 patients with tumors, 24 had astrocytomas of various grades, 3 had metastatic lesions, and an additional 3 had lymphomas. Computer-assisted stereotactic biopsy with arteriographic control is an accurate and relatively safe method of determining the histologic nature of any suspicious intracranial lesion. PMID- 2984486 TI - [Clinical features and application of enzyme immunoassay in a herpes zoster meningitis series]. PMID- 2984487 TI - [Portuguese-type familial amyloidotic polyneuropathy: presentation of a case]. PMID- 2984488 TI - [Metastasis of breast cancer and secondary acute myeloblastic leukosis. Simultaneous bone marrow infiltration]. PMID- 2984489 TI - Chemical and biochemical aspects of platelet activating factor: a novel class of acetylated ether-linked choline-phospholipids. PMID- 2984490 TI - GABA receptor agonists: pharmacological spectrum and therapeutic actions. AB - From the data discussed in this review it appears that GABA receptor agonists exhibit a variety of actions in the central nervous system, some of which are therapeutically useful (Table V). GABA receptor agonists, by changing the firing rate of the corresponding neurons accelerate noradrenaline turnover without changes in postsynaptic receptor density and diminish serotonin liberation with an up-regulation of 5HT2 receptors. These effects differ from those of tricyclic antidepressants which primarily block monoamine re-uptake and cause down regulation of beta-adrenergic and 5HT2 receptors. The GABA receptor agonist progabide has been shown to exert an antidepressant action which is indistinguishable from that of imipramine in patients with major affective disorders. The fact that: (a) GABA receptor agonists and tricyclic antidepressants affect noradrenergic and serotonergic transmission differently; and (b) tricyclic antidepressants alter GABA-related parameters challenges the classical monoamine hypothesis of depression and suggests that GABA-mediated mechanisms play a role in mood disorders. Decreases in cellular excitability produced by GABAergic stimulation leads to control of seizures in practically all animal models of epilepsy. GABA receptor agonists have a wide spectrum as they antagonize not only seizures which are dependent on decreased GABA synaptic activity but also convulsant states which are apparently independent of alterations in GABA-mediated events. These results in animals are confirmed in a wide range of human epileptic syndromes. GABA receptor agonists decrease dopamine turnover in the basal ganglia and antagonize neuroleptic-induced increase in dopamine release. On repeated treatment, progabide prevents or reverses the neuroleptic-induced up-regulation of dopamine receptors in the rat striatum and antagonizes the concomitant supersensitivity to dopaminomimetics. Behaviorally, GABA receptor agonists diminish the stereotypies induced by apomorphine or L-DOPA suggesting that GABAergic stimulation results also in an antidopaminergic action which is exerted beyond the dopamine synapse. These effects of GABA receptor agonists may represent the basis of the antidyskinetic action of these compounds which, however, remains to be fully confirmed. GABA receptor agonists reduce striatal acetylcholine turnover, an effect which occurs at doses much lower than those which affect dopamine neurons. Since hyperactivity of cholinergic neurons plays a determinant role in the pathogenesis of some parkinsonian symptoms, it is conceivable that GABAergic stimulation is effective in ameliorating Parkinson's disease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984491 TI - Subepithelial space in otitis media. AB - The role of the subepithelial space (SES) has not received sufficient attention in assessing pathogenesis, pathology, and therefore, clinical diagnosis and treatment of the various forms of otitis media (OM). Temporal bones from patients with OM were classified as cases of acute purulent (POM), serous (SOM), mucoid or secretory (MOM), or chronic otitis media (COM). Controlled morphometric studies were made of cellular components of the SES, along with studies of the epithelium and middle ear space. Corollary studies of biochemistry, cellular components, and prostaglandins (PGs) were done on fluid from the human middle ear. Middle ear effusions (MEE) from animal models of SOM, MOM, and POM were analyzed biochemically. Findings are surprising in that the SES was more actively involved in all forms of OM than had been thought, especially in MOM and COM. Implications are discussed. PMID- 2984492 TI - Adenoid cystic carcinoma of the nasopharynx after previous adenoid irradiation. AB - In 1978, Pratt challenged the otolaryngology community to identify an incidence of malignancy in individuals who have previously received radium therapy to the nasopharyngeal lymphoid tissues. This case report is a direct response to that quest and presents a well documented adenoid cystic carcinoma evolving 23 years after radium applicator treatment to the fossa of Rosenmuller. Although a cause and-effect relationship cannot be scientifically proven, the case history raises several important questions concerning the stimulating effects of radiation on the later onset of frank malignancy. PMID- 2984493 TI - Control of sigmoid sinus in glomus jugulare tumor resection. PMID- 2984494 TI - Re: Bell's palsy: the steroid controversy revisited. PMID- 2984495 TI - The diurnal variation of immunoreactive adrenocorticotropin in rhesus monkey plasma and cerebrospinal fluid. AB - ACTH immunoreactivity (ACTH-IR) in the plasma and cerebrospinal fluid (CSF) collected simultaneously from rhesus monkeys was found to undergo significant diurnal variations. In plasma, the mean peak ACTH-IR was 15.4 +/- 1.95 pg/ml at 0500 h, and the mean minimum concentration was 9.05 +/- 1.80 pg/ml at 1800 h. In CSF, the mean peak ACTH-IR concentration occurred at 1900 h and was 4.64 +/- 0.41 pg/ml. The mean minimum CSF ACTH-IR concentration was 2.93 +/- 0.26 pg/ml, occurring at 0500 h. This is the first report of a diurnal variation in CSF ACTH IR concentration and is consistent with other work suggesting that plasma ACTH and CSF ACTH originate from different sources. PMID- 2984496 TI - Preservation of human chorionic gonadotropin and placental lactogen secretion and normal morphology following long-term culture of normal human placental cells. AB - In order to study the regulation of hCG and hPL secretion during gestation, a system for the preservation of the functional integrity of normal placental cells in long-term culture was established. Normal term placental cells were dispersed with 0.25% trypsin-500 units DNAse I and cultured in a monolayer in Dulbecco's modified Eagle medium with 10% fetal bovine serum. Normal cell morphology, basal hCG and hPL production and hCG responses to dibutyryl cAMP were preserved till 54 days of culture. This model may be useful for the study of long-term regulation of normal placental hCG and hPL synthesis and secretion. PMID- 2984497 TI - Distribution of calmodulin in taste buds. AB - Calmodulin is higher in particulate fractions from bovine taste buds containing taste bud membranes which specifically bind sweet tastants compared to corresponding fractions from control non-taste bud bearing lingual epithelial tissue. As biochemical purity (i.e., membrane enzyme marker activity) of these membrane enriched fractions increased (P4B greater than P3B greater than P2B) calmodulin correspondingly increased (P4B greater than P3B greater than P2B); these increases also correlated with increased membrane purity as demonstrated by electron microscopy. All PB subfractions from taste buds contained a greater membrane concentration than those from PD subfractions and calmodulin was significantly increased in each corresponding subfraction. The presence of calmodulin in taste bud membranes, its correlation with membrane purification and reports that numerous drugs which induce taste loss are potent inhibitors of calmodulin suggest a role for calmodulin in taste function. PMID- 2984498 TI - Behavioral stress decreases plasma oxytocin concentrations in primates. AB - Using rhesus monkeys, we studied the effects of a behavioral stress on plasma concentrations of adrenocorticotropin, oxytocin, and vasopressin. The stress resulted in significant increases in adrenocorticotropin and significant decreases in oxytocin concentrations. No significant changes were seen in vasopressin concentrations. To further explore the relationship between plasma oxytocin and pituitary-adrenal function, dexamethasone was administered to rhesus monkeys. This resulted in significant increases in plasma oxytocin concentrations, while adrenocorticotropin decreased. PMID- 2984499 TI - Effects of psychoactive and non-psychoactive cannabinoids on neuroendocrine and testicular responsiveness in mice. AB - Repeated oral administration of the non-psychoactive cannabinol (CBN; 5 or 50 mg/kg) significantly reduced the concentration of norepinephrine (NE) in median eminence and greatly reduced NE levels 1 and 2 hrs after administration of alpha methylparatyrosine (alpha-MPT). The levels of dopamine (DA) in median eminence were significantly different, as indicated by the differences in slopes obtained in CBN- treated and control mice before and after alpha-MPT. Plasma levels of luteinizing hormone (LH) were significantly reduced in CBN-exposed mice before alpha-MPT, elevated at 1 hr post-injection, but were also reduced 2 hrs post injection at 50 mg/kg CBN. Follicle-stimulating hormone (FSH) levels were increased at 1 hr post-alpha-MPT in mice receiving 50 mg/kg CBN. Oral administration of CBN at 50 mg/kg for 4 days enhanced testicular testosterone (T) production in response to intratesticular in vivo injection of 2.5 or 25 mIU human chorionic gonadotropin (hCG). A single oral dose of the psychoactive delta 9-tetrahydrocannabinol (THC) enhanced the production of T 15 min after intratesticular LH (10 ng) injection. However, at 45 or 60 min post-THC treatment, the response to LH was significantly attenuated. These studies demonstrate that both psychoactive and non-psychoactive components of marihuana alter testicular responsiveness to gonadotropins in vivo. These effects may be biphasic, involving stimulation and inhibition of responsiveness, and appear to be correlated with alterations in plasma LH levels. Alterations in plasma gonadotropins may be mediated by cannabinoid effects on catecholamine concentrations in median eminence and THC-induced alterations in testicular responsiveness to gonadotropin probably also involve direct effects of THC at the gonadal level. PMID- 2984501 TI - Prolonged in vivo antagonism of central mu- and delta-opioid receptor activity by beta-funal trexamine. AB - beta-Funaltrexamine (beta-FNA) was tested in the spinal cord and supraspinally against inhibition of reflex bladder contractions produced in the anesthetized rat by the opioid-receptor selective agonists [D-Ala2, MePhe4, Gly (ol)5]enkephalin (DAGO, mu-agonist) and [D-Pen2, D-Pen5] enkephalin (DPDPE, delta agonist). All agents were microinjected either intracerebroventricularly (i.c.v.) or intrathecally (i.t.). beta-FNA (1-8 micrograms) produced long-lasting antagonism of both DAGO and DPDPE. Complete recovery from its effects was only observed some 24-32 h later. Higher doses of beta-FNA (4 and 8 micrograms i.t.) produced short-lived agonistic activity though the selectivity of this was not determined. It was concluded that beta-FNA was a potent, long-lasting antagonist at central opioid receptors in vivo but was unselective for the mu and delta opioid receptor. PMID- 2984500 TI - Enantiomers of [R,S]-thiorphan: dissociation of analgesia from enkephalinase A inhibition. AB - The [R] and [S] enantiomers of the enkephalinase A inhibitor [R,S]-thiorphan have been prepared by asymmetric synthesis. The [S] isomer is principally responsible for the angiotensin converting enzyme inhibitory activity of [R,S]-thiorphan, whereas there were only small differences in the ability of the [R] and [S] isomers to inhibit enkephalinase both in vivo and in vitro. In contrast, the in vivo analgesic activity of [R,S]-thiorphan resided principally in the [R] isomer. These data indicate a surprising dissociation of enkephalinase inhibition from analgesic activity. The fact that the two enantiomers of [R,S]-thiorphan were effective inhibitors of enkephalinase, yet the [R] isomer had substantially greater analgesic activity, indicates that factors other than enkephalinase inhibition may be important for [R, S]-thiorphan's analgesic properties. PMID- 2984502 TI - Characterization of central cholecystokinin receptors using a radioiodinated octapeptide probe. AB - We have developed a binding assay for 125I-Bolton-Hunter-labeled cholecystokinin octapeptide (125I-(BH)CCK8) using mouse cerebral cortex membrane preparations. This ligand interacts with cortical membrane preparations in a saturable, high affinity manner, satisfying the requirements for specific cholecystokinin receptor labeling. Salt is required for maximal binding and BSA is specifically inhibitory with cerebral cortical but not with pancreatic sites. Cholecystokinin peptides as small as CCK30-33 displace binding at low nanomolar concentrations. Dissociation of 125I-(BH)CCK8 is biphasic in both mouse and guinea pig cortex. Pretreatment of membranes at 37 degrees C results in a marked loss of recognition sites, suggesting that the sites may be rapidly metabolized in vivo. After 37 degrees C pretreatment, the loss of CCK recognition sites corresponds to a selective loss of the slow component of dissociation curves. This selective elimination of one dissociation population, as well as the biphasic dissociation kinetics, suggests that at least two distinct CCK receptor subtypes exist in the brain. PMID- 2984503 TI - Structural requirements for cannabinoid-induced antinociceptive activity in mice. AB - The antinociceptive activity of delta 9-THC was found to be 45 times greater after i.v. (ED50 = 1.0 mg/kg) than after s.c. administration while its ability to depress spontaneous activity was increased only 8 fold. Route of administration had less of an impact on the antinociceptive potency of hydroxylated metabolites and analogs of delta 9-THC. The structural requirements for antinociception and depression of spontaneous activity were found to be similar. Both routes of administration and structural alterations dramatically changed the antinociceptive potency of delta 9-THC. PMID- 2984504 TI - Changes in coenzyme Q level in mitochondria of cirrhotic rat liver. AB - In the cirrhotic rat liver induced by carbon tetrachloride and phenobarbitone, the concentrations of mitochondrial Coenzyme Q were measured in comparison with other respiratory components. The concentration of cytochrome a(+a3) and Coenzyme Q significantly increased in the cirrhotic liver, without any changes in the ratio of Coenzyme Q to cytochrome a(+a3). It is suggested that such increase of Coenzyme Q plays an important role as one of the adaptive responses to compensate for the prolonged metabolic overload on the mitochondrial respiratory assembly. Also, from the findings that the concentrations of cytochrome a(+a3) in the mitochondria of cirrhotic liver increase concomitant with the severity of cirrhosis, it is suggested that the rise of Coenzyme Q levels may be one of the indicators for the decreased functional reserve capacity in liver cirrhosis. PMID- 2984505 TI - Prenatal chlordiazepoxide effects on metrazol seizures and benzodiazepine receptors density in adult albino rats. AB - Adult offsprings of rats treated with daily injection of chlordiazepoxide (10 mg/kg, i.p.) during pregnancy showed a significant decrease in benzodiazepine receptors in the cortex and the cerebellum without apparent changes in receptor affinity. A reduced susceptibility to metrazol-induced epileptogenesis paralleled this change. These findings are discussed in relation to the differential vulnerability of various types of benzodiazepine receptors. PMID- 2984506 TI - Neurotensin is produced by and secreted from classic small cell lung cancer cells. AB - The presence of neurotensin in various human tumor cell lines was investigated by radioimmunoassay. High concentrations (0.06-5.1 pmol/mg protein) were detected in 50% of the classic but not variant small cell lung cancer or other human tumor cell lines examined. Biochemical studies indicated that the main peak of immunoreactivity coeluted with synthetic neurotensin using gel filtration and high pressure liquid chromatography techniques. Also, the rate of neurotensin secretion increased approximately 2-fold when theophylline was added which elevated intracellular levels of cAMP 4-fold. Because neurotensin is present in and secreted from many classic small cell lung cancer cells, it may function as a regulatory peptide in this disease. PMID- 2984507 TI - Dopaminergic and serotoninergic anorectics differentially antagonize insulin- and 2-DG-induced hyperphagia. AB - The efficacy of anorectic drugs has been studied in rats made hyperphagic by injection of insulin or of 2-deoxy-d-glucose (2-DG). It was found that anorectics that act through a serotoninergic mechanism, i.e., d- and d-l-fenfluramine, p chloroamphetamine, quipazine and fluoxetine antagonize both insulin- and 2-DG induced overeating, while anorectics acting through the dopaminergic system, i.e., d-amphetamine, diethylpropion, lisuride, bromocriptine and mazindol, antagonize the hyperphagia induced by 2-DG but not that induced by insulin. Neither serotoninergic nor dopaminergic anorectics modified insulin-induced hypoglycaemia. The serotonin (5-HT) receptor blocker metergoline did not modify the hyperphagic response to insulin or 2-DG. The present results indicate that there are different neuronal or humoral circuits underlying the hyperphagic responses to insulin and 2-DG. In addition, these results, which show different effectiveness of anorectic drugs depending on what has provoked the hyperphagia, suggest that differences in the etiology of the hyperphagia of obese subjects must be taken into consideration when choosing therapy. PMID- 2984508 TI - Novel adenosine receptors in rat hippocampus. Identification and characterization. AB - 2-Chloro[3H]adenosine, a stable analog of adenosine, was used to investigate the presence of adenosine receptors in rat hippocampal membranes that may mediate the depressant effects of adenosine on synaptic transmission in this tissue. Equilibrium binding studies reveal the presence of a previously undescribed class of receptors with a KD of 4.7 microM and a Bmax of 130 pmol/mg of protein. Binding is sensitive to alkylxanthines and to a number of adenosine-related compounds. The pharmacological properties of this binding site are distinct from those of the A1 and A2 adenosine receptors associated with adenylate cyclase. The results suggest that this adenosine binding site is a novel central purinergic receptor through which adenosine may regulate hippocampal excitability. PMID- 2984509 TI - Role of arginine vasopressin in control of ACTH and LH release during stress. AB - To determine the role of arginine vasopressin (AVP) in stress-induced release of anterior pituitary hormones, AVP antiserum or normal rabbit serum (NRS) was micro injected into the 3rd ventricle of freely-moving, ovariectomized (OVX) female rats. A single 3 microliter injection was given, and 24 hours later, the injection was repeated 30 min prior to application of ether stress for 1 min. Although AVP antiserum had no effect on basal plasma ACTH concentrations, the elevation of plasma ACTH induced by ether stress was lowered significantly. Plasma LH tended to increase following ether stress but not significantly so; however, plasma LH following stress was significantly lower in the AVP antiserum treated group than in the group pre-treated with NRS. Ether stress lowered plasma growth hormone (GH) levels and this lowering was slightly but significantly antagonized by AVP antiserum. Ether stress also elevated plasma prolactin (Prl) levels but these changes were not significantly modified by the antiserum. To evaluate any direct action of AVP on pituitary hormone secretion, the peptide was incubated with dispersed anterior pituitary cells for 2 hours. A dose-related release of ACTH occurred in doses ranging from 10 ng (10 p mole)-10 micrograms/tube, but there was no effect of AVP on release of LH. The release of other anterior pituitary hormones was also not affected except for a significant stimulation of TSH release at a high dose of AVP. The results indicate that AVP is involved in induction of ACTH and LH release during stress. The inhibitory action of the AVP antiserum on ACTH release may be mediated intrahypothalamically by blocking the stimulatory action of AVP on corticotropin-releasing factor (CRF) neurons and/or also in part by direct blockade of the stimulatory action of vasopressin on the pituitary. The effects of vasopressin on LH release are presumably brought about by blockade of a stimulatory action of AVP on the LHRH neuronal terminals. PMID- 2984510 TI - Affinity chromatography for human Na+, K+-ATPase inhibitors in plasma and urine. AB - Semi-purified dog kidney Na+,K+-ATPase cross-linked with ovalbumin was used in batch-wise affinity chromatography for the detection of endogenous Na+,K+-ATPase inhibitor in human plasma and urine. Ammonium acetate 1 M washed off the endogenous inhibitor from the immobilized enzyme. The inhibitory activity of the eluate from hypertensive plasma and urine was significantly higher (p less than 0.0025, n = 5 and p less than 0.005, n = 6 respectively) than that of normotensive. This latter was correlated with the ability of plasma from the same subjects to compete with ouabain binding to erythrocytes. Plasma and urine extracts inhibited the activity of Na+, K+-ATPase in a dose-dependent manner as ouabain does and were shown to contain 3 or 4 active compounds by high pressure liquid chromatography. The activity of some of these compounds was lost after peptidase treatment. These data support the heterogeneity of endogenous inhibitors of Na+,K+-ATPase activity in plasma and urine. PMID- 2984511 TI - [Radiotherapy of malignant salivary gland tumors]. AB - The results of treatment of 230 patients with different malignant tumors of the greater salivary glands (adenocystic carcinoma, adenocarcinoma, mucoepidermoid tumor, undifferentiated carcinoma, carcinoma in pleomorphous adenoma, epidermoid carcinoma, etc.) were analysed. Altogether 177 patients received combination treatment and 53 radiation therapy as a single modality. In preoperative irradiation the summary focal dose was 45-50 Gy, in postoperative irradiation 60 Gy, when radiation therapy was used alone the dose was 65-70 Gy. A study was made of the long-term therapeutic results with relation to a therapeutic modality, tumor morphological structure and stage. In combined therapy, the highest 5-year survival rates were in adenocystic carcinoma (83%) and mucoepidermoid tumor (78.5%), the lowest rate in undifferentiated carcinoma (16.6%). In radiation therapy used as a single modality the 5-year survival rates were 38.8% in adenocystic carcinoma and 8.3% in adenocarcinoma. PMID- 2984512 TI - [Radionuclide-thermographic diagnosis of neck paragangliomas]. AB - Nonchromaffin paragangliomas (chemodectomas) are rare tumors. Diagnosis of paraganglion tumors is difficult. The main diagnostic method is carotid angiography. Radionuclide thermographic studies were conducted in 16 neck paraganglioma patients for comprehensive diagnosis. Malignant tumors were found in 6 out of 16 patients. Paragangliomas of the carotid corpuscle were noted in 14 patients, those of the vagus nerve in 2 patients. In benign paraganglioma hyperthermia of the neck in the tumor region did not exceed 1.2 degrees C in 8 out of 10 patients. In all 6 patients with malignant paragangliomas, hyperthermia from 1.3 degrees and upwards was noted in the tumor region on the thermograms. On the scintigrams foci of increased RP accumulation were noted in 4 patients with malignant and in 2 patients with benign paragangliomas. The use of the radionuclide and thermographic methods makes it possible to extend the potentialities of differential diagnosis of benign and malignant neck paragangliomas. PMID- 2984513 TI - Endorphins: "opiates for the masses". AB - Endogenous opioid peptides rose to prominence with the discovery of the enkephalins in 1975. Since then, a vast amount of research has been directed toward understanding their role in normal and pathophysiological situations. Although the place of endogenous opioids in psychiatry remains uncertain, there is good evidence that a variety of tumors may secrete endorphins or enkephalins, and these may contribute to the non-metastatic complications of malignant disease. In addition, changes in cerebrospinal fluid met-enkephalin and beta endorphin after acupuncture may be involved in the effectiveness of this therapy in the treatment of heroin withdrawal and severe pain. The hormonal effects of opiate agonists and antagonists are now well characterized; exercise-induced changes in circulating catecholamines are markedly enhanced by the opiate antagonist naloxone. It is possible that the opiate inhibition of catecholamine release during exercise is a reflection of endogenous opioid modulation of effort perception. PMID- 2984514 TI - Control of hepatic glucose production during exercise. AB - Major advancements have occurred recently in study of the mechanisms of regulation of glycogenolysis and gluconeogenesis in perfused livers and isolated hepatocytes. Many questions remain unanswered, however, with respect to the control of these processes in the live exercising animal. Additional studies will be necessary to determine the relative roles of alpha- and beta-adrenergic receptor-mediated effects of circulating catecholamines, of glucagon, glucocorticoids, angiotensin II, vasopressin, insulin, and of the direct sympathetic innervation in regulation of hepatic glycogenolysis and gluconeogenesis during exercise. PMID- 2984516 TI - Exercise and endorphins--male responses. AB - The discovery of the endogenous opiates in the mid-1970's has led to a tremendous scientific effort attempting to determine the physiological role of these peptides. An increase in the peripheral plasma levels of beta-endorphin in humans after exercise has been noted by all investigators to date. This indication that the endogenous opiate system is activated during exercise should stimulate investigators to use exercise and training as models to aid in the understanding of these peptides. Unlike most other endocrines, plasma levels of beta-endorphin do not increase proportionally to work intensity. This conclusion is based on only one study and requires corroboration. All human studies to date have used radioimmunoassays, with one exception, and interestingly that study showed no consistent change in plasma levels of leucine enkephalin-like radioreceptor assayable ligands. In males, essentially no information is available concerning the effects of training on either acute or chronic responses to exercise. Studies using opiate antagonists (receptor type-specific) in human and animal models should prove useful in establishing or disproving roles for these peptides in appetite, pain perception, temperature regulation, metabolism, ventilation, and blood pressure control during exercise. PMID- 2984515 TI - Endorphins and exercise in females: possible connection with reproductive dysfunction. AB - Currently available measurements of beta-endorphin and beta-lipotropin in exercising women are in excellent agreement and indicate a 2-3 fold increase over basal levels. Possible effects of exercise upon the transfer of endorphins from the peripheral circulation to the brain are examined, and evidence is presented that suggests the occurrence of a concomitant exercise-related increase of endorphins in both humoral and central nervous system compartments. Steady-state measurements of circulating luteinizing hormone and follicle-stimulating hormone levels in oligo-amenorrheic athletes, on the other hand, do not agree. It is felt that the lack of consensus may be attributable partly to technical inadequacies and partly to lack of awareness of the need for frequent sampling. The bulk of the findings suggest a tendency for luteinizing hormone levels to be low and follicle-stimulating hormone levels to be normal or low, a pattern compatible with repeated activation of the CRH-ACTH-POMC system as a result of exercise. PMID- 2984517 TI - The role of calmodulin in hormone-stimulated lipolysis. AB - To explore the role of calmodulin (CaM) in lipolysis, studies were carried out on effects of CaM inhibitors on hormone-stimulated lipolysis, the activity of cAMP dependent protein kinase, and phosphorylation of endogenous substrate proteins. When adipocytes were incubated with trifluoperazine (TFP) and W-7 but not with W 5, stimulation of lipolysis by epinephrine was blunted. W-7 also inhibited lipolysis induced by ACTH, 1-methyl-3-isobutylxanthine (MIX) or (Bu)2 cAMP. The binding of 3H-cAMP to its receptor protein (the regulatory subunit of protein kinase) as well as the activity of cAMP-dependent protein kinase was suppressed by W-7, and the anti-CaM antibody, but not by W-5. The CaM-dependence of the protein kinase was also proved by the fact that the protein kinase activity that was markedly reduced in CaM-depleted cell extracts, was significantly restored by addition of exogenous CaM to them. Furthermore, W-7 decreased cAMP-stimulated phosphorylation of endogenous substrate proteins (mol wt 230k, 200k, 130k, 85k, 75k, and 50kdalton), among which the one of 85kdalton is most likely to be the hormone-sensitive lipase. These findings suggest that CaM is involved in the mechanism of hormone-induced lipolysis by exerting stimulatory effects on the activation of cAMP-dependent protein kinase in cell extracts capable of phosphorylating substrate proteins including hormone-sensitive lipase. PMID- 2984518 TI - Genetic analysis of Tn7 transposition. AB - The purpose of this work was to localize the DNA regions necessary for the transposition of Tn7. Several deletions of Tn7 were constructed by the excision of DNA fragments between restriction sites. The ability of these deleted Tn7s to transpose onto the recipient plasmid RP4 was examined. All the deleted Tn7s isolated in this work had lost their transposing capability. The possibility of complementing them was studied using plasmids containing all or part of Tn7. Two deleted Tn7s could not be complemented by an entire Tn7 indicating that a DNA sequence greater than the 42 bp terminal sequence is needed for recognition of the transposon by a transposition function. Four other deleted Tn7s could be complemented by Tn7. One of these was studied intensively in complementation experiments using different parts of Tn7 to obtain transposition. The results obtained allow us to propose that all genes needed for transposition of Tn7 onto plasmids are contained in a DNA segment of between 6.0 and 7.4 kb. Furthermore, one essential function must be contained in a DNA fragment longer than 2.5 kb on the right-hand end of Tn7. The classification of Tn7 with regard to the other transposable elements is discussed. PMID- 2984519 TI - Cloning and complementation analysis of the "Frizzy" genes of Myxococcus xanthus. AB - Fruiting-body formation in Myxococcus xanthus involves the aggregation of cells into raised mounds, where they sporulate. "Frizzy" mutants fail to aggregate into mounds, but rather aggregate into "frizzy" filaments (D.R. Zusman 1982). The frizzy mutations (frz) were found to be genetically linked. The region of DNA carrying the frz genes was cloned in Escherichia coli by selecting for the kanamycin resistance element present on a transposon Tn5 insertion linked to the frz genes. Phage P1 mediated transduction of the cloned DNA into M. xanthus frizzy mutants showed that the cloned DNA could complement the frz mutations. The cloned DNA was analyzed by isolating and characterizing new Tn5 insertions at short intervals within the M. xanthus DNA and by constructing in vitro deletions. The mutated DNA was then transduced into M. xanthus where the cloned DNA became integrated into the bacterial chromosome as gene replacements or as merodiploids. The gene replacement strains allowed us to define the limits of the frz region, since Tn5 insertions in the frz genes resulted in the frizzy phenotype. The merodiploid strains allowed us to perform complementation analyses. Using appropriate crosses, we were able to identify 5-6 frz complementation groups on 7.5 kb of cloned DNA. One of the complementation groups was separated from the others by 1.4 kb of DNA, whereas the others were contiguous. The different frz loci behave as separate transcriptional groups although interactions between some of the gene products are indicated. PMID- 2984521 TI - Kinetoplast DNA segments function as promoters in Escherichia coli cells. AB - By means of coupled transcription-translation in Escherichia coli cell-free system, an open reading frame was found in the Crithidia oncopelti maxi-circle kDNA segment cloned in the hybrid plasmid pCo1. Subfragments from this region were tested for their ability to function as promoters in E. coli cells. For this purpose the vector pVE8 was constructed using the aminoglycoside phosphotransferase II (APTII) gene from the Tn5 transposon, and the pHC79 cosmid. After cloning of Sau3A fragments of pCo1 by insertion into pVE8 three types of plasmids containing promoter sequences were obtained. Two of these plasmids displayed promoter strength in E. coli cells greater than that of the normal promoter of the APTII gene. However the promoters found are not necessary for the coupled transcription-translation of kDNA in the E. coli cell-free system. PMID- 2984520 TI - Activation of expression of a cloned archaebacterial gene in Escherichia coli by IS2, IS5, or deletions. AB - A DNA fragment from the methanogenic archaebacterium Methanococcus voltae, when cloned into the PstI site of the plasmid vector pBR322, complements the Escherichia coli argG mutation strongly or weakly depending on its orientation. Faster-growing variants derived from a strain containing the poorly expressed fragment were found to harbor plasmids which had undergone genetic rearrangements. Some of the plasmids were shown to have acquired an insertion element (IS2 or IS5), derived from the E. coli chromosome, close to the region essential for complementing activity. Other plasmids exhibited no homology with E. coli chromosomal DNA. These were found to represent multimeric forms of the parental plasmid in which 2-3 kb of DNA between the tet promoter and the argG complementing region had been deleted. Growth rates of the variant strains in the absence of arginine varied significantly, suggesting differences in efficiency of activation of the cloned DNA. PMID- 2984522 TI - Competence related proteins in the supernatant of competent cells of Bacillus subtilis. AB - We report that centrifugation at relatively high g-forces reduces the ability of competent cells of Bacillus subtilis to bind and take up DNA, and to be transformed. The centrifugation supernatant from competent cells restores this reduction of competence; the supernatant from non-competent cells is inactive. Phosphocellulose chromatography of centrifugation supernatants from radioactive competent cultures gave rise to six sharp peaks, together, these were shown by subsequent SDS polyacrylamide gel electrophoresis to contain over 60 different polypeptide bands. Peak II, which showed competence restoring activity, produced three polypeptides. When these bands were further examined, one of these exhibited DNA binding activity and the other two each contained a different endonuclease. Competence restoring activity was not recovered from the SDS polyacrylamide gel of peak II. The three peaks from non-competent cultures produced altogether five faint bands in gel electrophoresis. None of these bands were similar to those found in peak II. PMID- 2984523 TI - Additional genes essential for replication of the mini-F plasmid from origin I. AB - We isolated a series of Tn5-insertional mutants from the mini-F plasmid, which has a deletion in the origin II region and replicates exclusively from origin I, and found that the mutants that had Tn5 in either the F4 or the F5 gene were defective in their replication. It is concluded that, in addition to the F3 gene on which we have reported previously, both the F4 and the F5 genes are essential for the replication from origin I. PMID- 2984524 TI - Prolonged outbreak of Norwalk gastroenteritis in an isolated guest house. AB - During November and December 1982, a persistent outbreak of gastroenteritis took place in an isolated guest house. An estimated 26% of the changing population of guests and staff developed symptoms. Laboratory studies implicated Norwalk agent as the cause of illness, although the original source of the outbreak could not be established. Information on clinical features of the illness, its mode of spread, and eventual control is presented. PMID- 2984525 TI - Oncogenes. A possible role for cancer genes in human malignant disease. AB - A small class of normal cellular genes has the potential to transform cells malignantly. These cellular oncogenes have been identified by their similarity to the transforming genes of oncogenic RNA viruses (retroviruses), and by their ability, under certain circumstances, to transform cells into which they have been transferred. Cellular oncogenes are not normally tumorigenic and, indeed, appear to be critically involved in normal cellular processes, possibly at the level of growth regulation. However, it appears that a number of mechanisms exist whereby these oncogenes can become malignantly activated, leading to uncontrolled cellular proliferation. The evidence implicating cellular oncogenes in human malignant disease is reviewed. PMID- 2984526 TI - Screening for AIDS. PMID- 2984527 TI - Mapping of the deletion in the genome of HSV-1 strain HFEM responsible for its avirulent phenotype. AB - To determine the exact map position of the deletion in the genome of HSV-1 strain HFEM, the Bam HI DNA fragment B of the HSV-1 strains HFEM, F, and of HSV-R-HFehx C19 were cloned in the Bam HI site of the bacterial plasmid vector pAT153 and analysed in detail. Analysis of the insert of the three recombinant plasmids harbouring the Bam HI DNA fragment B of HSV-1 strain F (pHSF-B-B), HSV-1 strain HFEM (pHSHF-B-B), and recombinant HSV-R-HFehx-C19 (pHSR19-B-B) was performed using a variety of restriction enzymes and Southern blot hybridization. These comparative studies revealed that the HpaI DNA fragment P of the intact viral genome of HSV-1 strain F is not present in the genome of HSV-1 strain HFEM, indicating that the deletion in HSV-1 strain HFEM corresponds to this fragment, which spans the coordinates 0.762 to 0.790 of the HSV-1 DNA. The size of the deletion was found to be 4.1 kbp, corresponding to 2.7 Md. PMID- 2984528 TI - Oncology seen through the prism of Wilms tumor. PMID- 2984530 TI - Changing patterns of acquired immunodeficiency syndrome in hemophilia patients- United States. PMID- 2984529 TI - Benign metastatic islet cell tumour of the pancreas. AB - A nine-year-old girl was fortuitously found to have a symptomless metastatic islet cell tumour (apudoma) of the head of the pancreas, the first to be described in a child. The tumour has not changed in size over 3 years and the patient has shown normal growth and development. Initial chemotherapy had no effect. PMID- 2984531 TI - Prevalence of cytomegalovirus excretion from children in five day-care centers- Alabama. PMID- 2984532 TI - Performance of laboratories in testing for rabies virus--United States. PMID- 2984533 TI - [Infiltration of bile duct carcinoma along the wall]. AB - The characteristics of the infiltration of bile duct carcinoma along the wall were studied from 25 cases with resection. The histologic mode of the spread of carcinoma was the mucosal infiltration in 1 case, mucosal and transmural infiltration in 2 cases, transmural infiltration in 22 cases. The average distance from the gross margin of the tumor to the margin of spread of growth in the bile duct as determined microscopically was 19.1 mm in the direction of the liver (the longest was 85 mm), and 5.8 mm toward the duodenum (the longest was 20 mm). The distance of the polypoid or nodular type of the gross appearance of the tumor is shorter than that of the infiltrating type. The polypoid type showed inverted U-type in obstructive or stenotic feature of bile duct radiologically and the infiltrating type showed U-type, V-type or constricted type. The distance of the tubular adenocarcinoma in histological cellular type tended to be longer than that of papillary or undifferentiated type. The severer the invasion of the carcinoma to the lymphatic vessels was, the longer the distance was. PMID- 2984535 TI - [Pituitary-adrenocortical response to surgical stress in male patients]. AB - Effects of major surgical stress on the plasma levels of twelve kinds of steroid hormones, ACTH and renin activity, and on the excretion rates of urinary free cortisol and acid labile aldosterone etc. were studied in eleven male patients for seven days following operation. In another groups of patients, ACTH tests were carried out in order to investigate the effect of ACTH on the adrenal steroid hormone production. Also, an in vitro assay of the effect of ACTH on the adrenal tissue was performed. Surgical stress provoked marked increase in plasma levels of cortisol, despite little change in aldosterone levels. The postoperative plasma testosterone concentration showed a profound decrease throughout the observation period. The plasma renin activity reached the highest level on the first postoperative day, when plasma aldosterone was at the lowest level. It was concluded that the postoperative changes in the plasma steroid hormone levels were markedly influenced by other physical and humoral disorders provoked with surgical stress. PMID- 2984534 TI - [A new method for preparation of islet cells from dogs]. AB - A new method for preparation of viable islet cells from the normal pancreas is introduced. After total pancreatectomy, the pancreatic duct is cannulated and perfused with Hanks' balanced salt solution (HBSS) containing 0.2% collagenase at 37 degrees C for 30-45 minutes. The gland is then chopped and dissociated by shaking in a water bath. The cell suspension is filtered through steel mesh and washed with cold HBSS by centrifugation. By this method, collagenase is applied preferentially into the acinar tissue and the islet yield is greatly enhanced with decreased acinar contamination. Estimation of insulin and amylase in pancreatic tissue and islet rich cell suspension (graft) indicates a 57% islet recovery and a six-fold enrichment in islet concentration. Graft prepared by this method is autotransplanted into totally pancreatectomised dogs and normoglycemia is achieved in ten of 13 animals. PMID- 2984536 TI - [Bilateral breast carcinoma]. AB - Out of 4,777 cases of female breast carcinoma operated at the Cancer Institute Hospital, Tokyo from 1946 to 1980, 170 cases, 3.56% had bilateral breast carcinoma. According to our criteria, 157 cases (92. 3%, 3.29%) were classified as primary and 103 cases (7.6%, 0.27%) were metastatic. Our criteria depend upon the histological proof whether intraductal cancer parts in the smaller or in the second cancer exist or not. Out of 157, 48 cases (1.01%) were simultaneous (within one year) and 109 cases (2.28%) were subsequent bilateral carcinoma cases. The clinical characteristics of primary bilateral breast carcinoma were earlier onset, earlier stage and better prognosis than those of unilateral cases. From the epidemiological point of view, bilateral cases showed high percentage in sterile case and familial disposition. In so called breast cancer family, bilateral cases were common and the tendency of earlier onset in succeeding generation was recognized. The annual incidence rate of bilateral breast carcinoma among amputated patients for unilateral carcinoma was calculated as 4%. So it is well said that the incidence of breast carcinoma among Japanese women is as low as one-sixth of that of Americans, but the rate of bilaterality is high and almost the same as Americans. PMID- 2984537 TI - [A case of membranous obstruction of inferior vena cava at the hepatic associated with hepatocellular carcinoma and liver cirrhosis]. AB - A case of inferior vena cava obstruction at the hepatic portion associated hepatocellular carcinoma with and liver cirrhosis is reported, which was treated with lateral segmentectomy of the liver after transcatheter angioplasty. A 36 year-old male, who had noticed venous dilatation in the abdominal wall and legs from his childhood, visited a doctor complaining of right upper quadrate pain and was diagnosed liver cirrhosis. One year later ultrasonography revealed a liver tumor, which was diagnosed as hepatocellular carcinoma by ultrasonically guided aspiration cytology. Inferior and superior vena cavography revealed complete membranous obstruction of inferior vena cava at the hepatic portion with marked collateral circulation through azygos, hemiazygos and phrenic veins. The caval pressure difference between above and below the obstruction was 16.5 cm H2O. The membranous obstruction was perforated and dilated by transluminal angioplasty using Dotter's balloon catheter. The obstructive segment of inferior vena cava changed into 8mm in diameter after the second angioplasty, and the caval pressure difference between above and below the stenosis decreased to 10 cm H2O. Lateral segmentectomy of the liver was performed. Histopathologic diagnosis was clear cell type hepatocellular carcinoma with liver cirrhosis. Marked postoperative liver damage was observed and transcatheter caval dilatation was performed again. The pressure of inferior vena cava below the stenosis decreased to 8 cm H2O. One year and 8 months after the operation, the patient is healthy without recurrence of cancer. PMID- 2984539 TI - Potential energy functions and the role of the conformational entropy of clonidine-like imidazolidines in determining their affinity for alpha-adrenergic receptors. AB - Energy as a function of the ring interplanar torsional angle was calculated for 12 2-(phenylimino)imidazolidines by the method of perturbation configuration interaction using localized orbitals. The potential energy functions indicate that the molecules may assume any conformation within rather broad limits. The functions were used in calculating the gas phase and solution conformational entropies. The latter were used as the independent variable in regression analysis to derive equations connecting the conformational entropy with pKi (pKi = -log Ki) for [3H]clonidine displacement (literature data). As a comparison, correlations between pKi and several other parameters (modified neglect of diatomic overlap-computed highest occupied and lowest unoccupied molecular orbital energies and dipole moments; pKa; log P; substituent steric parameters) were sought. Correlation coefficients C greater than 0.6 were obtained with the conformational entropy (-0.77), and the ortho steric parameters (-0.71). The correlation with the conformational entropy was not markedly improved by adding other parameters in multiple regression analysis. This result is discussed in terms of the contribution to the ligand-receptor complexation free energy arising from the conformational restriction of the ligands upon binding to the receptor. PMID- 2984538 TI - Cannabinoid inhibition of adenylate cyclase. Biochemistry of the response in neuroblastoma cell membranes. AB - The inhibition of adenylate cyclase activity by cannabimimetic compounds in a membrane fraction from cultured neuroblastoma cells has been examined. The inhibition was shown to be concentration-dependent over a nanomolar range for both delta 9-tetrahydrocannabinol and its synthetic analog, desacetyllevonantradol. Inhibition was rapid and reversible. The cannabimimetic compounds caused a decrease in Vmax of the enzyme, with no alteration in the Km for substrate. The effects of these compounds were related to the ability of the enzyme to be regulated by divalent cations and guanine nucleotides. The inhibition was greatest at micromolar Mg2+ or Mn2+ concentrations and was abolished at less than 1 mM MnCl2. In the hormone-stimulated state, the enzyme appeared to be regulated by one Mg2+ site. The addition of cannabimimetic or muscarinic cholinergic agents transformed the enzyme into one in which more complex regulation by divalent cations was observed. Half-maximal inhibition of adenylate cyclase was observed at 800 nM GTP for both cannabimimetic and muscarinic cholinergic agents. The substitution for GTP of a nonhydrolyzable analog resulted in activation of the enzyme and failure to respond to either class of inhibitory agents. If the Mg2+ concentration was reduced and exposure to the GTP analog was of short duration, inhibition by both cannabimimetic and muscarinic agents could be observed in the presence of forskolin. This study points to the similarities between the enzyme inhibition by cannabimimetic compounds and by muscarinic cholinergic compounds. It is inferred that the cannabimimetic compounds must act via regulatory mechanisms similar to those operating for receptor-mediated inhibition of adenylate cyclase. PMID- 2984540 TI - Interaction of DNA-intercalating antitumor agents with adrenoceptors. AB - The interaction between some examples of mononuclear and binuclear DNA intercalating antitumor agents and alpha- and beta-adrenoceptors has been studied using radioligand-binding assays. Competition for 125I-BE 2254, [3H]rauwolscine, and (-)-[3H]dihydroalprenolol binding was used to assess affinity for alpha 1-, alpha 2-, and beta-adrenoceptor-binding sites, respectively. Two homologous series of alkyl-linked diacridines and diquinolines were found to interact poorly with beta-adrenoceptors, with only the largest members having appreciable affinity. By contrast, these compounds bind strongly and in a complex manner to alpha 1- and alpha 2-adrenoceptors. The affinity of diacridines for both alpha adrenoceptor classes has a parabolic dependence on alkyl chain length with the hexyl and pentyl derivatives being the most potent at the alpha 1- (Ki = 11.5 +/- 2.3 nM) and alpha 2- (Ki = 143 +/- 26 nM) binding sites, respectively. The dependence of inhibition constants on linker chain length for the diquinolines is more complicated, with the ethyl- and heptyl-linked dimers having the greatest affinity for each alpha subclass. There is a nadir in affinity for the pentyl and butyl ligands and an increase in dissociation constant for octyl and longer homologues. Thus, the ethyl diquinoline has Ki values of 6.6 +/- 1.2 and 110 +/- 14 nM for the alpha 1- and alpha 2- adrenoceptors, respectively, and, correspondingly, the heptyl derivative has values of 39 +/- 4 and 51 +/- 1 nM. These findings are discussed with respect to a model of the alpha-adrenoceptor in which the radioligand-binding site is situated in a trench or cleft, surrounded by a flat surface bounded by walls. Daunomycin was found to have no affinity for adrenoceptors of any type and mitoxantrone similarly fails to interact with alpha 2- and beta-adrenoceptors, but binds to the alpha 1 subclass with an inhibition constant (Ki) of 3930 +/- 420 nM. Bisantrene also has no affinity for beta adrenoceptors but binds to alpha 1- and alpha 2- adrenoceptors with Ki values of 145 +/- 24 and 2310 +/- 430 nM, respectively. Among the mononuclear acridine drugs studied, only nitracrine shows detectable interaction with beta adrenoceptors (Ki = 760 +/- 50 nM). This compound, like bisantrene, has high affinity for the alpha 1-adrenoceptor (Ki = 131 +/- 17 nM) and moderate affinity for the alpha 2 subclass (Ki = 2180 +/- 500 nM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984541 TI - The interaction between calcium and the activation of Na+, K+-ATPase by noradrenaline. AB - The interaction of noradrenaline, various cation chelators and calcium on Na+, K+ ATPase from rat cerebral cortex plasma membranes was studied. It was shown that chelation of inhibitory cations by EGTA, EDTA and dipyridyl activated Na+, K+ ATPase to the same extent as noradrenaline but at higher concentrations; increasing concentrations of EGTA depressed the activation by noradrenaline; calcium in the form of a calcium-EGTA buffer depressed Na+, K+-ATPase at physiological concentrations; the inhibition of Na+, K+-ATPase by calcium is dependent on the magnesium concentration in the assay and the inhibition by calcium was partially reversed by noradrenaline. PMID- 2984542 TI - Multiple forms of synthase D phosphatase and phosphorylase a phosphatase in liver and regulatory effects of metabolites on their activities. AB - The smooth endoplasmic reticulum (ER) and cytosol fractions of liver homogenates exhibit phosphoprotein phosphatase activity towards glycogen synthase D and phosphorylase a. The following observations suggest that liver contains multiple forms of these phosphatases. Synthase phosphatase activity in either fraction was more readily inactivated by heating than phosphorylase phosphatase activity. Both synthase phosphatase and phosphorylase phosphatase activities in smooth ER were non-competitively inhibited by Mg2+, but were activated by this ion in the cytosol. Synthase phosphatase activities in cytosol and smooth ER were stimulated by a number of sugar phosphates, particularly glucose-1-phosphate, galactose-6 phosphate and fructose-6-phosphate. Erythrose-4-phosphate stimulated synthase phosphatase activity in the cytosol, but inhibited the microsomal enzyme. Phosphorylase phosphatase activities in either fraction were inhibited by most sugar phosphates. Adenosine mono-, di- and tri-phosphates inhibited phosphatase activities in both fractions. Low concentrations of AMP and ADP inhibited phosphorylase phosphatase activities to a greater extent than synthase phosphatase activities. Chromatography of the smooth ER fraction on DEAE cellulose resulted in the separation of synthase phosphatase from phosphorylase phosphatase, as soluble proteins. The elution profile for the microsomal phosphatase was different from that for the cytosol enzymes. It is concluded that: both synthase phosphatase and phosphorylase phosphatase in liver have at least two isoenzyme forms; synthase phosphatase and phosphorylase phosphatase are separate enzymes; the different behaviour of microsomal and cytosol phosphatases towards divalent cations and sugar phosphates provides a potential mechanism for the differential regulation of these activities in liver. PMID- 2984543 TI - Permeability changes resulting from virus-cell fusion: temperature-dependence of the contributing processes. AB - A new assay for membrane fusion, using the fluorescent probe pyrene-sulphonyl phosphatidyl ethanolamine, has been developed. Fusion between the envelope of Sendai virus and human erythrocytes or Lettre cells has a Q10 of approximately 4 at 37 degrees C, increasing to approximately 7 at 7 degrees C; there is no lag to onset of fusion. Viral neuraminidase has a Q10 of 2.3 between 37 degrees C and 4 degrees C. Its action limits the extent of fusion by causing the elution of virus; this effect is particularly marked at low temperature because of the difference in Q10 for fusion and neuraminidase. The temperature-dependence of the initiation of permeability changes following the removal of inhibitory amounts of Ca2+ is approximately 2; thus membrane fusion is the principal temperature sensitive step during the permeabilization of cells by Sendai virus. A recovery process, by which cells become insensitive to the removal of Ca2+ and which therefore limits the extent of permeabilization, has a Q10 of 7.4 between 37 degrees C and 21 degrees C. It is concluded that the lag to onset of permeability changes is not due to a lag in virus-cell membrane fusion, but to the gradual acquisition of a threshold level of membrane damage; the extent of permeabilization depends on the rate of fusion relative to the rates of neuraminidase and recovery. PMID- 2984544 TI - [The use of computers in molecular biology studies]. AB - The paper contains a brief review of the studies involving application of computers, carried out at the Institute of Molecular Biology (investigation of the structure and dynamics of nucleic acids; modelling of the DNA-ligand interactions; analysis of the spectral data of different types: electron paramagnetic resonance, magnetic circular dichroism, absorption and fluorescence spectra; analysis of the thin-layer samples labelled by radioactive elements; chromosome analysis). PMID- 2984545 TI - [Activation of adenylate cyclase system enzymes and lysosomal acid phosphatase in target cells interacting with T-killer cells]. AB - The interaction of T-killers with target cells was studied to reveal the biochemical changes in the latter. On specific binding of target cells with T killers the activity in target cells of cAMP phosphodiesterase increased 2.1 fold, the level of cAMP decreased 1.5-fold, the adenylate cyclase activity decreased 2.0-fold, the phosphorylation of intracellular proteins decreased 1.8 fold, the cAMP-dependent protein kinase activity decreased 1.7-fold. No change in the activity of lysosomal enzymes was observed. At the "independent target cells lysis" stage the level of cAMP increased 1.8-fold, the phosphodiesterase activity decreased 1.7-fold, the cAMP-dependent protein kinase activity increased 1.8 fold, the released activity of acid phosphatase increased up to 40% compared with the control cells. In the presence of 1 mM dibutyryl cAMP the released activity of the acid phosphatase in target cells was inhibited by 29%, the target cells lysis was decreased by 23,5%. The data obtained allowed to suppose that the activation of the host lysosomal enzymes causes target cells autolysis and that cAMP takes part in the regulation of these processes. PMID- 2984546 TI - [Specific protection of DNA by distamycin A, netropsin and bis-netropsins against the action of DNAse I]. AB - Interaction of netropsin, distamycin A and a number of bis-netropsins with DNA fragments of definite nucleotide sequence was studied by footprinting technique. The nuclease protection experiments were made at fixed DNA concentration and varying ligand concentrations. The affinity of ligand for a DNA site was estimated from measurements of ligand concentration that causes 50% protection of the DNA site. Distribution pattern of the protected and unprotected regions along the DNA fragment was compared with the theoretically expected arrangement of the ligand along the same DNA. The comparison led us to the following conclusions: 1. Footprinting experiments show that at high levels of binding the arrangement of netropsin molecules along the DNA corresponds closely to the distribution pattern expected from theoretical calculations based on the known geometry of netropsin- DNA complex. However, the observed differences in the affinity of netropsin for various DNA sequences is markedly greater than that expected from theoretical calculations. 2. Netropsin exhibits a greater selectivity of binding than that expected for a ligand with three specific reaction centers associated with the antibiotic amide groups. It binds preferentially to DNA regions containing four or more successive AT pairs. Among 13 putative binding sites for netropsin with four or more successive AT pairs there are 11 strong binding sites and two weaker sites which are occupied at 2 D/P less than or equal to 1/9 and 2 D/P = 1/4, respectively. 3. The extent of specificity manifested by distamycin A is comparable to that shown by netropsin although the molecule of distamycin A contains four rather than three amide groups. At high levels of binding distamycin A occupies the same binding sites on DNA as netropsin does. 4. The binding specificity of bis-netropsins is greater than that of netropsin. Bis netropsins can bind to DNA in such a way that the two netropsin-like fragments are implicated in specific interaction with DNA base pairs. However, the apparent affinity of bis-netropsins estimated from footprinting experiments is comparable with that of netropsin for the same DNA region. 5. At high levels of binding bis netropsins and distamycin A (but not netropsin) can occupy any potential site on DNA irrespectively of the DNA sequence. 6. Complex formation with netropsin increases sensitivity to DNase I at certain DNA sites along with the protection effect observed at neighboring sites. PMID- 2984547 TI - [Immunoglobulin kappa-genes. Cloning, hybridization and structural analysis]. AB - The principal problems in molecular and genetic immunology to be resolved are the structure of Ig-genes and the regulation of their expression. The isolation of mRNA for light and heavy Ig-chains would be a first step along this line. A combination of two approaches may be the best strategy in mRNA preparation. Affinity purification allows one to obtain pure mRNA in a one-step procedure whereas immunoprecipitation makes it possible to prepare mRNAs for both heavy and light immunoglobulin chains in considerable amounts. In the series of experiments, conditions for synthesis of long and short cDNA chains were elaborated and the clone containing the fragment of kappa-chain gene was isolated and characterized. This clone was used as a probe for hybridization with genomic DNA from myeloma and hybridoma cells. It was shown that both allelic genes on homologous chromosomes were rearranged in myeloma MOPC21 cells. The original cell line of hybridoma PTF02 contains the embryonic gene as well as the differentiated genes. However, only differentiated genes can be detected in a similar experiment conducted with the same hybridoma after passage on mice. In conclusion, the coordination of homologous and heterologous chromosome expression in B cells in discussed in terms of the feed-back control. PMID- 2984548 TI - [Problems of cellular activity regulation by secondary messengers]. AB - The paper surveys the data obtained during the last years in the Laboratory of Regulation of Cellular Activity of the Institute of Molecular Biology. The data mostly concern the enzymes taking part in metabolism and in biological action of the secondary messengers, as well as the mechanisms of regulation of some metabolic pathways in the cultured cells. The results pertinent to the mechanisms of chromatin condensation and based on this genetic control processes are also discussed. PMID- 2984549 TI - [A system of EcoRV restriction-modification: genes, enzymes and synthetic substrates]. AB - The genes, encoding the restriction endonuclease and modification methylase EcoRV have been cloned from the natural plasmid pLG13 into pBR32 derivative vector pIL233. A resultant clone, expressing both enzyme activities, was used as a source of DNA for sequencing these genes by a procedure, that employed construction of deletion derivatives used to locate borders (by means of a functional test) and to sequence ca. 300 bp near the deletion breakpoint. From the sequence data, we infer that the endonuclease, a 29 KDa protein, and the methylase, a 36 KDa protein, are transcribed from a 310 bp intergenic region in opposite directions. There is no apparent homology between the enzymes and genes of the EcoRI and the EcoRV systems. A synthetic decamer, containing the EcoRV endonuclease recognition sequence and a phosphoamide bond at the cleavage point, is not cleaved by the highly purified endonuclease; the unmodified synthetic decamer is cleaved at the same conditions, only that the cleavage occurs to produce a blunt end--GAT/ATC, and not in a place previously reported (GATAT/C). PMID- 2984550 TI - [Various novel properties of transcriptionally active mini-chromosomes of the SV40 virus]. AB - The treatment of isolated SV40 mini-chromosomes by DNA-topoisomerase I leads to relaxation of DNA within a small fraction (2-5%) of mini-chromosomes strongly enriched in endogenous RNA-polymerase. The DNA supercoiling in the bulk of mini chromosomes remained unchanged. The relaxable fraction proved to be specifically hypersensitive to DNAase I, but lost hypersensitivity after prior topoisomerase treatment. The DNA relaxation induced either by topoisomerase or DNAase I nicking and breaking led to almost a complete loss of proteins from this fraction while the DNA-protein interactions in the bulk of mini-chromosomes remained unchanged. Endogenous RNA-polymerase remained specifically enriched in these uncoated mini chromosomes. It is concluded that (1) there is an elastic torsional strain in DNA within transcriptionally active mini-chromosomes, (2) DNA-protein interactions are altered within transcriptionally active mini-chromosomes, (3) there is evidence to indicate that local DNA conformational transitions in transcriptionally active chromatin are caused by DNA torsional strain. PMID- 2984552 TI - Transcriptional and post-transcriptional control of PHO8 expression by PHO regulatory genes in Saccharomyces cerevisiae. AB - A DNA fragment bearing the PHO8 gene, which encodes repressible alkaline phosphatase of Saccharomyces cerevisiae, was cloned. Northern hybridizations with the PHO8 DNA as probe indicated that the PHO8 transcript is 1.8 kilobases in length and is more abundant in cells grown in low-phosphate medium than in high phosphate medium. The pho9 mutant, whose phenotype is defective in the activity of repressible alkaline phosphatase, produced as much of the PHO8 transcript as did the PHO9+ cells. Hence, the PHO9 product should act at the post transcriptional level. The pho4 mutant could not derepress the PHO8 transcript, whereas the pho80 mutant could, irrespective of the amount of Pi in the medium, as has been suggested by genetic study. PMID- 2984551 TI - A domain of methylation change at the albumin locus in rat hepatoma cell variants. AB - A well-differentiated rat hepatoma cell line, Fu5-5, yields variant clones whose rate of secretion of serum albumin ranges from 40 to less than 0.08 micrograms of albumin/mg of cell protein per 48 h. Clones were classified as high producers (10 to 40 micrograms/mg per 48 h), intermediate producers (1 to 10 micrograms/mg per 48 h), low producers (0.1 to 1.0 micrograms/mg per 48 h), and null variants (less than 0.1 micrograms/mg per 48 h). Albumin synthetic rates are proportional to secretion rates and range from 0.9 to less than 0.002% of total protein synthesis as measured by pulse-labeling. Steady-state albumin mRNA levels were measured by filter hybridization of fragmented, end-labeled mRNA and by Northern blotting. Message levels are proportional to albumin synthetic rates except for a high producer in which albumin mRNA is less elevated than the synthetic rate. The extent of methylation was quantitated at each of 24 CpG-containing sites or site clusters at the albumin locus. These sites span a region that contains the albumin gene as well as 10 kilobases of the 5' flank and 1 kilobase of the 3' flank. An 8-kilobase region is described, with boundaries in the 5' flank and in the middle of the gene, within which all 11 sites examined showed a correlation of undermethylation with the high-producer phenotype. In contrast, 12 of 13 sites outside of this region showed no phenotype correlation. Null variants derived from a high producer underwent de novo methylation of this domain. Six independent hybrid clones derived from the cross of a high producer with a null variant showed extinction of albumin production and hypermethylation of the domain. Apparently these cells retain the capacity for the de novo methylation of these specific sites. PMID- 2984553 TI - Stable transcription complex on a class III gene in a minichromosome. AB - We have constructed recombinant simian virus 40 molecules containing Xenopus 5S RNA and tRNA genes. Recombinant minichromosomes containing these genes were isolated to study the interaction of RNA polymerase III transcription factors with these model chromatin templates. Minichromosomes containing a tRNAMet gene can be isolated in a stable complex with transcription factors (IIIB and IIIC) and are active in vitro templates for purified RNA polymerase III. In contrast, minichromosomes containing a 5S RNA gene are refractory to transcription by purified RNA polymerase III in either the absence or the presence of other factors. PMID- 2984554 TI - Concerted evolution of dispersed Neurospora crassa 5S RNA genes: pattern of sequence conservation between allelic and nonallelic genes. AB - About 100 genes coding for 5S RNA in Neurospora crassa are dispersed throughout the genome (Selker et al., Cell 24:815-818, 1981; R. L. Metzenberg, J. N. Stevens, E. U. Selker, and E. Morzycka-Wroblewska, manuscript in preparation). The majority of them correspond to the most abundant species (alpha) of 5S RNA found in the cell. Gene conversion, gene transposition, or both may be responsible for the maintenance of sequence homogeneity (concerted evolution) of alpha-type 5S genes. To explore these possibilities, we isolated and characterized separate 5S regions from two distantly related laboratory strains of N. crassa. Restriction and sequence analyses revealed no differences in molecular location of allelic 5S genes between the two strains. However, the DNA sequences around the 5S genes are ca. 10% divergent. We concluded that transposition is not frequent enough to account for the concerted evolution of N. crassa alpha-5S genes. In contrast to sequence divergence in the flanking regions between the two strains, the 5S transcribed regions are identical (with one exception), suggesting that these genes are being corrected. We have found that flanking sequences of various N. crassa 5S genes within each strain are largely different. Thus, if the correction mechanism is based on gene conversion, it is limited to the transcribed regions of the genes. However, we did find a short region of consensus including the sequence TATA located 25 to 30 nucleotides preceding the position of transcription initiation. This region may be involved in the transcription of N. crassa 5S genes. PMID- 2984556 TI - Nonreciprocal exchanges of information between DNA duplexes coinjected into mammalian cell nuclei. AB - We have examined the mechanism of homologous recombination between plasmid molecules coinjected into cultured mammalian cells. Cell lines containing recombinant DNA molecules were obtained by selecting for the reconstruction of a functional Neor gene from two plasmids that bear different amber mutations in the Neor gene. In addition, these plasmids contain restriction-length polymorphisms within and near the Neor gene. These polymorphisms did not confer a selectable phenotype but were used to identify and categorize selected and nonselected recombinant DNA molecules. The striking conclusion from this analysis is that the predominant mechanism for the exchange of information between coinjected plasmid molecules over short distances (i.e., less than 1 kilobase) proceeds via nonreciprocal homologous recombination. The frequency of homologous recombination between coinjected plasmid molecules in cultured mammalian cells is extremely high, approaching unity. We demonstrate that this high frequency requires neither a high input of plasmid molecules per cell nor a localized high concentration of plasmid DNA within the nucleus. Thus, it appears that plasmid molecules, once introduced into the nucleus, have no difficulty seeking each other out and participating in homologous recombination even in the presence of a vast excess of host DNA sequences. Finally, we show that most of the homologous recombination events occur within a 1-h interval after the introduction of plasmid DNA into the cell nucleus. PMID- 2984555 TI - Role of specific simian virus 40 sequences in the nuclease-sensitive structure in viral chromatin. AB - A nuclease-sensitive region forms in chromatin containing a 273-base-pair (bp) segment of simian virus 40 DNA encompassing the viral origin of replication and early and late promoters. We have saturated this region with short deletion mutations and compared the nuclease sensitivity of each mutated segment to that of an unaltered segment elsewhere in the partially duplicated mutant. Although no single DNA segment is required for the formation of a nuclease-sensitive region, a deletion mutation (dl45) which disrupted both exact copies of the 21-bp repeats substantially reduced nuclease sensitivity. Deletion mutations limited to only one copy of the 21-bp repeats had little, if any, effect. A mutant (dl135) lacking all copies of the 21- and 72-bp repeats, while retaining the origin of replication and the TATA box, did not exhibit a nuclease-sensitive region. Mutants which showed reduced nuclease sensitivity had this effect throughout the nuclease-sensitive region, not just at the site of the deletion, indicating that although multiple determinants must be responsible for the nuclease-sensitive chromatin structure they do not function with complete independence. Mutant dl9, which lacks the late portion of the 72-bp segment, showed reduced accessibility to BglI, even though the BglI site is 146 bp away from the site of the deletion. PMID- 2984558 TI - [Gianotti-Crosti syndrome. HBsAG-negative papular acrodermatitis, an infantile papulovesicular acrolocalized syndrome]. AB - We report three cases of the Gianotti-Crosti syndrome (papulovesicular-acro located syndrome of childhood), a self-limited common disease of childhood, which may be caused by various viral infections. The hepatitis B virus, however, is excluded by definition as a causal agent. The syndrome is characterized by itching papulo-vesicular skin lesions measuring 1-5 mm in diameter and localized to the limbs and face. The clinical and terminological differences between the HBsAG negative Gianotti-Crosti syndrome and the HBsAG positive Gianotti disease are emphasized. PMID- 2984557 TI - Method of mapping DNA replication origins. AB - We have developed a method which allows determination of the direction in which replication forks move through segments of chromosomal DNA for which cloned probes are available. The method is based on the facts that DNA restriction fragments containing replication forks migrate more slowly through agarose gels than do non-fork-containing fragments and that the extent of retardation of the fork-containing fragments is a function of the extent of replication. The procedure allows the identification of DNA replication origins as sites from which replication forks diverge. In this paper we demonstrate the feasibility of this procedure, with simian virus 40 DNA as a model, and we discuss its applicability to other systems. PMID- 2984559 TI - [Experimental morphologic study of changes in the myocardium in alcoholic intoxication in the rat]. PMID- 2984560 TI - Role of the umuC gene in postreplication repair in UV-irradiated Escherichia coli K-12 uvrB. AB - The role of the umuC gene product in postreplication repair was studied in UV irradiated Escherichia coli K-12 uvrB cells. A mutation at umuC increased the UV radiation sensitivities of uvrB, uvrB recF, uvrB recB, and uvrB recF recB cells; it also increased the deficiencies in the repair of DNA daughter-strand gaps in these strains, but it did not affect the repair of DNA double-strand breaks that arose from unrepaired DNA daughter-strand gaps. We suggest that the umuC gene product is involved in a minor system for the repair of DNA daughter-strand gaps, possibly the repair of overlapping DNA daughter-strand gaps. PMID- 2984561 TI - Genetic control of heat resistance and thermotolerance by recA and uvrA in E. coli K12. AB - Several recA and uvrA derivatives of E. coli K12 AB1157 develop a transient increase in heat resistance, i.e. induced thermotolerance after a brief exposure to 43.5 degrees C (less than 1 h). Thermotolerance was identified from the appearance of an inflection in the survival curve or from the loss of heat resistance in the presence of chloramphenicol (CAM) or rifampicin. Heat resistance and induced thermotolerance were enhanced by recA and uvrA gene functions and their contribution was roughly as follows: AB1157 (recA+ uvrA+) greater than AB2463 (recA- uvrA+) greater than AB1886 (recA+ uvrA-) greater than AB2480 (recA- uvrA-). In heat resistance, uvrA and recA contributed approximately equally and their effects were additive. Induced thermotolerance developed sooner and was maintained at a higher level in the presence of uvrA as compared with recA. Since uvrA-dependent excision repair is scheduled prior to recA-dependent (postreplication) repair, induction of thermotolerance may be linked to DNA repair. Although recA and uvrA play a distinct role, they are not essential, and thermotolerance can develop in the absence of either one or both of these gene functions. Furthermore, since thermotolerance can be induced in recA mutants (AB2463 and AB2480), its biochemical pathway must be different from that of the recA-dependent SOS system. PMID- 2984562 TI - A unique type of cyclic AMP-binding protein of Trypanosoma cruzi. AB - On centrifugation on sucrose density gradients, the cyclic AMP-receptor protein of Trypanosoma cruzi was clearly resolved from the type II regulatory subunit of protein kinase from bovine heart (S20,W = 8.25 and 4.1, respectively). The binding of cyclic [3H]AMP to these two proteins was affected to different extents by several cyclic AMP analogues. Such differences between the cyclic AMP-receptor protein of T. cruzi and cyclic AMP-binding proteins of other eukaryotes might be exploitable by chemotherapy. PMID- 2984563 TI - Ketoconazole in the management of precocious puberty not responsive to LHRH analogue therapy. AB - Three boys 3.3 to 3.9 years old, who had precocious puberty that was unresponsive to an analogue of gonadotropin-releasing hormone, were treated with the antifungal agent ketoconazole for up to 12 months. Within 48 hours the serum testosterone concentration fell to normal in two boys and was significantly reduced in the third, paralleling major improvements in behavior. Reciprocal changes in serum levels of 17-hydroxyprogesterone suggested that C17-20 lyase was the principal site of drug action. Although there was evidence of a blunted cortisol reserve during the first week of treatment, the cortisol response to ACTH1-24 had returned to normal by one month of continuous treatment, and normal diurnal cortisol rhythm was preserved. No adverse clinical or biochemical side effects were noted during 9 to 12 months of continuing treatment. During that time, growth velocity was significantly reduced in all three boys, from a mean rate of 1.5 +/- 2.0 cm per year before treatment to 5.9 +/- 0.6 cm per year after ketoconazole therapy. There was a simultaneous retardation of the rate of skeletal maturation. The striking improvements in behavior were sustained for the duration of treatment. These preliminary data suggest that administration of ketoconazole may be a satisfactory treatment for precocious puberty in boys and possibly for other conditions characterized by androgen excess. PMID- 2984565 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 17-1985. A 13-year-old boy with aniridia and proteinuria 11 years after nephrectomy for a Wilms' tumor. PMID- 2984564 TI - Decreased receptor binding of biologically inactive thyrotropin in central hypothyroidism. Effect of treatment with thyrotropin-releasing hormone. AB - Previous studies have suggested that certain cases of idiopathic central hypothyroidism of hypothalamic origin may result from the secretion of biologically inactive thyrotropin. To investigate this possibility and to define the mechanism of defective hormone action, we measured the adenylate cyclase stimulating bioactivity (B) and receptor-binding (R) activity of purified immunoreactive serum thyrotropin (I) from seven patients with hypothalamic hypothyroidism. We found a strikingly decreased R/I ratio (less than 0.15) in patients as compared with controls (0.6 to 2.7) and a similarly decreased B/I ratio (less than 0.2 vs 2.8 to 5.6). After acute injection of thyrotropin releasing hormone (TRH, 200 micrograms intravenously), the R/I ratio increased in two of three patients, but the B/I ratio became normal in only one. After administration of TRH for 20 to 30 days, an increase in immunoreactive serum thyrotropin was observed in all patients. Moreover, both ratios returned to normal in all but one patient, who had apparent desensitization. The increase in the amount and bioactivity of secreted thyrotropin after long-term TRH therapy resulted in enhanced secretion of serum thyroid hormones in all patients studied. We conclude that in certain cases of hypothalamic hypothyroidism, secreted thyrotropin lacks biologic activity because of impaired binding to its receptor; TRH treatment can correct both defects. These data suggest that TRH regulates not only the secretion of thyrotropin but also its specific molecular and conformational features required for hormone action. PMID- 2984566 TI - Risk of transmission of HTLV-III by needle stick. PMID- 2984568 TI - Screening donated blood and plasma for HTLV-III antibody. Facing more than one crisis? PMID- 2984567 TI - Epstein-Barr virus-associated B-cell proliferations of diverse clonal origins after bone marrow transplantation in a 12-year-old patient with severe combined immunodeficiency. AB - A 12-year-old boy with severe combined immunodeficiency who had been kept in a gnotobiotic environment since birth received bone marrow from a histoincompatible sibling in an attempt to reconstitute immunologic function. To prevent graft versus host disease, the donor's marrow was treated in vitro with monoclonal antibody and complement to remove alloreactive T cells. Eighty days after transplantation, the patient had a systemic illness characterized by fever, thrombocytopenia, gastrointestinal pain, and bleeding; he died on the 124th post transplantation day. Postmortem examination revealed multiple tumor-like B-cell proliferations, recipient in origin, in numerous organs. Epstein-Barr virus (EBV) was isolated from the patient's pharyngeal secretions; EBV nuclear antigen was found in spontaneously transformed peripheral-blood lymphocytes, inflammatory cells from peritoneal fluid, and bone marrow cells; and EBV genomes were discovered in all tumor tissues. The donor's serum showed evidence of past EBV infection. Analysis of cellular immunoglobulin and immunoglobulin gene DNA from the tumors indicated both monoclonal and oligoclonal B-cell proliferations. These findings provide evidence for the evolution of EBV-induced polyclonal activation of B cells to oligoclonal B-cell proliferation and finally to monoclonal B-cell lymphoma. PMID- 2984569 TI - Stimulation of the Na/H exchanger of sea urchin eggs by phorbol ester. AB - On fertilization of a sea urchin egg, marked changes occur in the cytoplasmic concentration of calcium and hydrogen ions. These ionic signals represent the necessary and sufficient stimuli for the increased metabolism, protein synthesis and DNA synthesis that constitute egg activation. Cytoplasmic alkalinization, the major immediate cause of the increased rate of protein synthesis which occurs at fertilization, arises because the sperm-induced intracellular calcium transient activates a coupled flux of sodium ions and hydrogen ions across the oolemma. The experiments reported here suggest that the second messenger which links the activation of the Na/H exchange to the calcium transient may be a substance which stimulates protein kinase C8, as 12-O-tetradecanoyl phorbol acetate (TPA), a known activator of protein kinase C9, appears to stimulate protein synthesis by turning on the Na/H exchanger and causing a cytoplasmic alkalinization. Our data indicate that one consequence of treating other tissues with TPA, a tumour promoter, may be an increase in intracellular pH. PMID- 2984570 TI - Focus formation in rat fibroblasts exposed to a tumour promoter after transfer of polyoma plt and myc oncogenes. AB - The gene encoding the large-T protein of polyoma virus (plt), the E1A genes of adenoviruses, the viral myc gene (v-myc) or rearranged forms of the cellular c myc gene confer on rat embryo fibroblast (REF) cells in primary culture a series of new properties ('immortality', reduced serum requirement and sensitivity to transformation by viral and activated cellular oncogenes) but do not induce the appearance of transformed foci. We now report that focus formation can be induced after transfer of these genes into either REF or established FR3T3 rat cells by subsequent exposure to the tumour promoter 12-O-tetradecanoylphorbol 13-acetate (TPA). Frequencies of transformation are in the same range as those usually observed for transformation with complete polyoma DNA or with a mixture of cloned myc and ras oncogenes. These results further characterize the 'immortalized' state induced by plt and myc as one in which the cells maintain a normal growth control in many respects but can be further acted upon to produce a neoplastic progeny. PMID- 2984571 TI - Tissue-specific expression of rat myosin light-chain 2 gene in transgenic mice. AB - One approach to determining how the differential expression of specific genes is regulated in higher organisms is to introduce cloned copies of the genes (or parts of the genes) into the genomes of individual organisms from the very beginning of their development. The way in which the exogenous genetic information behaves during the development of the experimental organisms can then provide a means of defining the DNA sequences that restrict the expression of the gene to specific cell types and times of development. So far, several different genes have been introduced into the genomes of mice, but in only a few cases have the exogenous genes retained the tissue specificity of expression of the equivalent endogenous genes. I report here that in two out of three 'transgenic' mice carrying copies of the rat gene for skeletal muscle myosin light chain 2, the exogenous gene is expressed specifically in skeletal muscle cells. The sequences contained in the cloned copy of the myosin light-chain 2 gene used in these experiments are thus sufficient to confer a tissue-specific pattern of expression. PMID- 2984572 TI - Regulation of a collagen gene promoter by the product of viral mos oncogene. AB - Oncogenic transformation of cells produces important changes in the biosynthetic pattern of certain cellular proteins. For example, the synthesis of type I collagen in transformed fibroblasts is severely reduced as a result of changes in transcription. Here we report the results of DNA-mediated transfection experiments using recombinant plasmids in which the promoter region of the alpha 2(I) collagen gene is fused to an easily recognizable marker gene, and cell lines expressing the marker gene are isolated. Our data show that the expression of the marker gene fused to the cloned alpha 2(I) collagen promoter is strongly inhibited by v-mos transformation, suggesting that a common mechanism inhibits both the transfected and endogeneous alpha 2(I) collagen promoters. PMID- 2984573 TI - Activation of protein kinase C potentiates isoprenaline-induced cyclic AMP accumulation in rat pinealocytes. AB - The pineal gland has proven to be an excellent model for the study of adrenergic control systems. Noradrenaline, released from sympathetic nerve terminals in the pineal gland, regulates a large nocturnal increase in melatonin synthesis by stimulating the activity of arylalkylamine N-acetyltransferase (NAT, EC 2.3.1.87) 30-70-fold. An essential step in both the induction and maintenance of high NAT activity is an increase in intracellular cyclic AMP. Noradrenaline acts via beta adrenoceptors to increase pineal cyclic AMP by activating adenylate cyclase, and the activation of pineal alpha 1-adrenoceptors potentiates beta-adrenergic stimulation not only of NAT but of both cyclic AMP and cyclic GMP. Here we describe investigations designed to test whether alpha 1-adrenergic potentiation of beta-adrenergic stimulation of pineal cyclic AMP involves protein kinase C. Our results suggest that kinase activation is involved and the data provide the first demonstration of a synergistic interaction between Ca2+-phospholipid dependent protein kinase (protein kinase C) and neurotransmitter-dependent stimulation of cyclic AMP. PMID- 2984574 TI - Human granulocyte-macrophage colony-stimulating factor is a neutrophil activator. AB - The polymorphonuclear leukocyte (PMN), or neutrophil, is the major host defence cell protecting the body against invasion by bacteria and fungi. Products of oxidative metabolism mediate PMN microbicidal and tumoricidal activity but the mechanisms by which these pathways become activated are not well understood. We have previously described a human granulocyte-macrophage colony-stimulating factor (GM-CSF) of relative molecular mass (Mr) 22,000 that also inhibits neutrophil motility (NIF-T activity). Because of its direct action on granulocytes, this lymphokine is a candidate for a neutrophil-activating factor. We have studied the effect of GM-CSF/NIF-T on superoxide anion generation in response to the bacterial chemo-attractant N-formylmethionyl-leucylphenylalanine (f-MLP), and report here that PMNs preincubated with either purified natural GM CSF or biosynthetic (recombinant) GM-CSF showed increased (as much as fourfold) superoxide anion production in response to f-MLP. These results indicate that human GM-CSF is a neutrophil-activating factor. PMID- 2984575 TI - Isolation of a cDNA clone corresponding to an X-linked gene family (XLR) closely linked to the murine immunodeficiency disorder xid. AB - The striking number of human and murine immunodeficiency disorders which map to the X chromosome suggests that genes localized on this chromosome must have important roles in lymphocyte development. At least seven distinct disorders in the human and two in the mouse disrupt lymphocyte maturation, particularly that of B cells, at characteristic stages. As functional genes mapping to the X chromosome in one mammal are found on the X chromosome in all other mammals, the same genes regulating lymphocyte development are expected to be found on the X chromosome in mouse and man. Investigations into the possible mechanisms of these X-linked disorders have been hampered by the lack of molecular probes for the genes or gene products affected; because of this, and the possibility of correlating one or more of the several hundred B- or T-cell-specific genes with a specific mutation, we surveyed 15 different B- and T-cell-specific cDNA clones for localization to the X chromosome. We report here the characterization of one of these murine cDNA clones, which hybridizes with a large, X-linked gene family, designated XLR (X-linked, lymphocyte-regulated). We show that the XLR gene family is closely linked to the X-linked immunodeficiency described in the CBA/N mouse strain (xid), by restriction fragment length polymorphism (RFLP) analysis of DNA from mice congeneic for xid. This finding, together with data on the expression of the XLR locus in B cells, indicates that this gene family either includes the locus defined by the xid mutation or is adjacent to it in a gene complex which may be important in lymphocyte differentiation. PMID- 2984576 TI - Activation of major histocompatibility complex class I mRNA containing an Alu like repeat in polyoma virus-transformed rat cells. AB - Class I genes of the major histocompatibility complex (MHC) appear to be activated in mouse cells transformed by the DNA tumour virus simian virus 40 (SV40). Conversely, suppression of MHC class I genes has been reported in adenovirus-12-transformed baby kidney rat cells. We have now investigated the expression of genes encoded by the rat MHC locus in rat fibroblast cells transformed by polyoma virus (Py). Using a mouse genomic H-2 clone as a probe in Northern transfer hybridization analysis, we have observed a high level of expression of rat MHC class I messenger RNA in all the transformed rat cell lines analysed. The class I 1.6-kilobase (kb) mRNA activated in Py-transformed rat cells appears to contain an Alu-like type II repeat element, as the same 1.6-kb mRNA is detected using either the H-2 class I sequence or a repetitive Alu-like type II element as a probe. High levels of heterogeneous poly(A)+ transcripts of 0.5-0.8 kb are also observed in Py-transformed rat cells using probes containing an Alu-like type II repetitive element. PMID- 2984577 TI - Cigarette smoke induces DNA single-strand breaks in human cells. AB - Epidemiological evidence suggests that smoking is a major cause of human lung cancer. However, the mechanism by which cigarette smoke induces the cancer remains obscure, although in tobacco carcinogenesis, promotion and/or co carcinogenesis may have crucial roles. The epidemiological data show that if an individual stops smoking, the risk of his contracting lung cancer increases no further. Moreover, laboratory experiments show that cigarette smoke condensate (CSC) exhibits co-carcinogenic and promoting activities in tumour production and malignant transformation. Clastogenic action is thought to be intimately involved in tumour promotion, and it is therefore interesting that visible chromosome changes such as chromosome aberrations and sister chromatid exchanges are known to be caused by cigarette smoke. However, there has been no previous direct demonstration that cigarette smoke can cause single-strand breaks (SSB) in DNA. Here we report that cigarette smoke induces considerable numbers of DNA SSB in cultured human cells, and that such strand breaks may be ascribed to active oxygen generated from cigarette smoke. PMID- 2984578 TI - Clusters of CpG dinucleotides implicated by nuclease hypersensitivity as control elements of housekeeping genes. AB - DNA sequences of the X-chromosome-linked hypoxanthine phosphoribosyltransferase (HPRT) and glucose 6-phosphate dehydrogenase (G6PD) genes have revealed the presence of clusters of CpG dinucleotides, raising the possibility that such clusters are involved in the control of expression of these genes, which are expressed in all tissues. Although CpG clusters are not exclusive features of the X chromosome, the analysis of X-linked genes provides the means to determine whether CpG clusters are control elements; one of the two homologous X loci in female mammals is not expressed, so that active and inactive versions of the gene can be compared. In fact, it has been shown that these CpG clusters are undermethylated when the gene is active and extensively methylated when the gene is inactive. In addition to hypomethylation, chromatin hypersensitivity to endonuclease digestion is a known hallmark of regulatory sequences in eukaryotic genes. We report here that the CpG clusters of the active hprt and g6pd genes are not only undermethylated, but also hypersensitive to MspI, DNase I and S1 nuclease, further supporting the suggestion that they are involved in the control of expression of these genes. PMID- 2984579 TI - Specific interaction between phosphatidylinositol 4,5-bisphosphate and profilactin. AB - There is evidence that the polymerization of actin takes place at the plasma membrane, and that profilactin (profilin/actin complex), the unpolymerized form of actin found in extracts of many non-muscle cells, serves as the immediate precursor. Both isolated profilin and profilactin interact with detergent when analysed by charge shift electrophoresis, indicating that they have amphipathic properties and may be able to interact directly with the plasma membrane. We demonstrate here that isolated profilin, as well as the profilactin complex, interacts with anionic phospholipids. Phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) was found to be the most active phospholipid, causing a rapid and efficient dissociation of profilactin with a concomitant polymerization of the actin in appropriate conditions. These and other observations suggest the possibility of a relationship between the induction of actin filament formation and the increased activity in the phosphatidylinositol cycle seen as a result of ligand-receptor interactions in various systems. PMID- 2984580 TI - The anxiogenic action of RO 5-4864 in the social interaction test: effect of chlordiazepoxide, RO 15-1788 and CGS 8216. AB - RO 5-4864 (20 mg/kg), a benzodiazepine with high affinity for peripheral-type benzodiazepine binding sites in rat kidney and brain, but not for the "classical" CNS sites, reduced the time spent by pairs of rats in active social interaction, without reducing locomotor activity, possibly reflecting an anxiogenic action. This anxiogenic effect was not reversed by chlordiazepoxide (5 or 10 mg/kg) given acutely, but was reversed by chlordiazepoxide (5 mg/kg) given for 5 days prior to testing. RO 15-1788 (10 mg/kg), a drug that antagonises several effects of benzodiazepines but has little affinity for peripheral-type sites, had no action on the reduction in social interaction induced by RO 5-4864. However, CGS 8216 (10 mg/kg) which also antagonises the effects of benzodiazepines and has little affinity for RO 5-4864 recognition sites, significantly enhanced the reduction in social interaction caused by RO 5-4864, and the combination produced a significant decrease in locomotor activity. These results are discussed in terms of possible sites of action of RO 5-4864 on the GABA-benzodiazepine receptor complex. PMID- 2984581 TI - Lesioning of serotoninergic and noradrenergic nerve fibres of the rat brain does not decrease binding of 3H-clonidine and 3H-rauwolscine to cortical membranes. AB - alpha 2-Adrenoceptors located presynaptically on nerve terminals are known to modulate the release of neurotransmitters from noradrenergic and serotoninergic neurons. The pre- and/or postsynaptic localization of binding sites for alpha 2 adrenergic radioligands, the agonist 3H-clonidine and the antagonist 3H rauwolscine, was investigated in the rat cerebral cortex by the use of specific neurotoxins. Intracerebroventricular injections of 6-hydroxydopamine (6-OH-DA) and 5,7-dihydroxytryptamine (5,7-DHT) were used to destroy the noradrenergic and serotoninergic neurons, respectively, and the success of the treatment was controlled by measurement of tritium accumulation in cortex slices incubated with 3H-noradrenaline or 3H-serotonin. In cortical membranes, 3H-rauwolscine bound to a single site (KD about 5 nmol/l; Bmax 217-247 fmoles/mg protein), whereas 3H clonidine bound to a high affinity site (KD 0.6-1.4 nmol/l) and a low affinity site (KD 6-10 nmol/l). The total number of high plus low affinity 3H-clonidine binding sites was about two thirds of the number of 3H-rauwolscine binding sites. 6-OH-DA treatment significantly increased the number of high affinity 3H clonidine binding sites without reducing the number of high plus low affinity binding sites, indicating a denervation supersensitivity. KD- as well as Bmax values for 3H-rauwolscine remained unaltered after 6-OH-DA-treatment. Since an increase in postsynaptic alpha 2-adrenoceptors due to 6-OH-DA-administration might have masked a loss of presynaptic alpha 2-adrenergic binding sites, rats were chronically treated with high doses of clonidine in order to prevent a possible supersensitivity of postsynaptic receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984582 TI - In vivo demonstration of alpha-2-adrenoceptor-mediated inhibition of the excitatory non-cholinergic neurotransmission in guinea pig airways. AB - In anaesthetized guinea pigs, vagal nerve stimulation caused an atropine- and hexamethonium-resistant increase in the respiratory insufflation pressure. Clonidine (0.05 mg/kg i.v.) inhibited the increase in insufflation pressure caused by vagal stimulation. This effect of clonidine was antagonized by the alpha-2-antagonist yohimbine (0.5 mg/kg i.v.), but not by the alpha-1-antagonist prazosin (0.03 mg/kg i.v.). In conclusion, the increase in the respiratory insufflation pressure, caused by stimulation of noncholinergic nerves, seemed to be controlled by inhibitory alpha-2-adrenoceptors in guinea pig airways. PMID- 2984584 TI - A kinetic study of the release of noradrenaline by electrical stimulation: influence of presynaptic alpha-adrenoceptors. AB - Dog saphenous vein strips were incubated with 1.4 mumol/l 3H-(--)-noradrenaline for 60 min, after inhibition of the noradrenaline-metabolizing enzymes and of extra-neuronal uptake. At the end of the incubation period the strips were perifused for 150 min; cocaine (10 mumol/l) was added to the perifusion fluid from t = 75 min onwards. In some experiments either phentolamine (10 mumol/l) or clonidine (0.1 mumol/l) was also added at this time. Some strips were subjected to electrical stimulation from t = 100 to 150 min of perifusion (t = 0 being the start of perifusion), with frequencies ranging from 0.5 to 13.5 Hz. A compartmental analysis of spontaneous or electrically-induced efflux of 3H noradrenaline was made. The spontaneous efflux had a long half time (t/2 = 124 min) and most of the 3H-noradrenaline which had accumulated in the strips did not participate in the efflux ("bound fraction", representing 90% of tissue activity at t = 100 min of perifusion). Neither phentolamine nor clonidine modified the half time or the "bound fraction" observed for spontaneous efflux. Electrical stimulation (greater than 0.5 Hz) mobilized only one compartment of noradrenaline, which represented about 50% of the noradrenaline accumulated in the strips. The half time of 3H-efflux induced by electrical stimulation decreased when the frequency increased from 0.5 Hz up to 13.5 Hz. Phentolamine increased the rate of efflux for all frequencies of stimulation and decreased the half time of efflux. However, the releasable pool of noradrenaline was only increased by phentolamine at 0.5 Hz, but not at higher frequencies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984583 TI - Mechanism of alpha 2-adrenergic inhibition of neuroeffector transmission in the mouse vas deferens. AB - The process by which the activation of presynaptic alpha 2-adrenoceptors inhibits the release of noradrenaline from terminals of postganglionic sympathetic nerves was studied in the mouse isolated vas deferens. Clonidine was used as a prototypic agonist. Field stimulation-evoked excitatory junction potentials (e.j.p.s) were recorded from individual muscle cells. The e.j.p. amplitudes were taken as a measure of transmitter release. Changes in the external Ca2+ concentration from 2.5 to 1.25 or 5 mM caused corresponding changes in the size of e.j.p.s. When the normal Ca2+ concentration of the medium (2.5 mM) was substituted by equimolar quantities of Ba2+ or Sr2+, the e.j.p. amplitudes decreased considerably. Clonidine (0.3-30 nM) inhibited the nerve stimulation evoked e.j.p. amplitudes in a concentration-dependent manner, without altering appreciably the frequency of spontaneous e.j.p.s. Procedures known to enhance Ca2+ entry into nerve terminals, like a high Ca2+ medium (Ca2+ 5 mM) or 4 aminopyridine 30 microM reduced the effect of clonidine. Repetitive nerve stimulation at 3 Hz, which is supposed to lead to an accumulation of free Ca2+ inside nerve terminals, similarly counteracted the effect of clonidine 10 nM. Whereas the alpha 2-adrenergic inhibition of the first e.j.p. in a train was unaffected, the inhibition of all successive e.j.p.s was gradually decreased. At 5 mM Ca2+ only the time-course of facilitation became faster, the decrease in alpha 2-adrenergic inhibition proceeded with the same pulse-dependent rate as at a normal external Ca2+ concentration, although from a lower initial level.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984585 TI - Studies on the mechanism of clonidine-induced mydriasis in the rat. AB - Intravenous administration of clonidine hydrochloride (3-100 micrograms/kg) produced a dose-dependent pupillary dilation in anaesthetized rats. All experiments were carried out in rats in which vagosympathetic nerve trunks were sectioned bilaterally at the cervical level. Clonidine-induced mydriasis was present only in those preparations having intact parasympathetic neural tone to the iris. Depletion of CNS monoamines by more than 95% with reserpine (5 mg/kg) and alpha-methyl-para-tyrosine (2 X 300 mg/kg) failed to alter the dose-response relation to clonidine. Pretreatment with the alpha-2-adrenoceptor antagonist, yohimbine hydrochloride (1.5 mg/kg), produced about a 10-fold shift to the right in the pupillary dose-response curve to clonidine. Yohimbine administered after the highest dose of clonidine also antagonized the mydriatic response. The above results suggest that clonidine acts on CNS post-synaptic alpha-2-adrenoceptors to produce mydriasis by withdrawal of parasympathetic neural tone to the iris. In an attempt to assess the physiological substrate(s) involved, mydriatic responses, due to parasympatho-inhibition, were evoked by electrical stimulation of ascending (sciatic nerve and medullary) and descending (hypothalamic) pathways. Yohimbine (0.3 and 1.0 mg/kg) produced a dose-dependent inhibition of the pupillary dilation evoked by stimulation of the sciatic nerve and medullary loci, whereas these doses of yohimbine failed to alter the dilation in response to hypothalamic stimulation. Similarly, monoamine depletion greatly antagonized the pupillary dilation elicited by sciatic nerve and medullary stimulation without significantly affecting mydriasis due to hypothalamic stimulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984586 TI - Demonstration of Ri-type adenosine receptors in bovine myocardium by radioligand binding. AB - Adenosine has been shown to have negative inotropic, chronotropic and dromotropic effects on the heart. The pharmacological profiles of these effects suggest that they are mediated via Ri (A1) adenosine receptors, but a direct demonstration of these receptors is still missing. In the present study we report direct labelling of these receptors with (-)N6-[125I]-p-hydroxyphenylisopropyladenosine [( 125I]HPIA)1. The radioligand bound in a saturable and reversible manner to a crude membrane preparation, the Bmax-value was 30.5 fmol/mg protein and the KD value 1.1 nmol/l. A similar affinity of the ligand was obtained in kinetic and competition experiments. Competition experiments with a variety of adenosine analogues gave a pharmacological profile characteristic of Ri adenosine receptors with high affinities of N6-substituted derivatives and a marked stereospecificity for N6-phenylisopropyladenosine (PIA). Purification of the membrane preparation by density gradient centrifugation resulted in a 30-fold increase in the number of binding sites which was paralleled by a similar increase in the number of binding sites for [3H]ouabain. Guanine nucleotides decreased binding of [125I]HPIA in a dose-dependent manner, but the IC50-values were considerably higher than those reported in other tissues. Finally, binding of [125I]HPIA appeared to be entropy-driven which has been shown to be characteristic of agonist binding to Ri adenosine receptors. These results suggest the presence of Ri adenosine receptors in ventricular myocardium which may be responsible for the mediation of the effects of adenosine and its analogues. PMID- 2984587 TI - Forskolin-induced elevation of rat jejunal cyclic AMP levels and stimulation of active glucose transport in vitro. AB - The adenylate cyclase stimulator forskolin enhanced both mucosal cyclic AMP levels and D-aldohexose transport in rat jejunum in vitro in a concentration dependent manner. With mucosal cyclic AMP elevated to 400% of basal values, active D-glucose transport was at 200% of control values. Transport parameters, as determined in a three compartment model using a dual label method, indicate a) increased "uphill" glucose transport at the brush border membrane with higher intracellular accumulation, b) unchanged serosal to mucosal glucose fluxes and passive permeabilities. Transport stimulating effects of forskolin were also present with D-galactose in the rat. In mouse jejunum active transport of D glucose and 3-O-methylglucose was increased by forskolin 10 mumol/l. Phlorizin inhibited D-glucose transport as well as L-glucose transport in the rat were not changed by the persisting cyclic AMP elevation induced by forskolin. The results show a positive correlation of active intestinal hexose transport with a modest elevation of mucosal cyclic AMP. In accordance to current models of cyclic AMP induced changes in intestinal fluid absorption, rat jejunal net fluid transport was reduced by 40% with 20 mumol/l forskolin. PMID- 2984588 TI - beta-Adrenoceptor subtypes in sections of rat and guinea-pig kidney. AB - The present autoradiographical study examines the distribution of the two beta adrenoceptor subtypes in sections of rat and guinea-pig kidney. The radioligand [125Iodo]-(--)-cyanopindolol was used for the labelling of beta-adrenoceptors and the selective beta-adrenoceptor blocking agents ICI 89-406 (beta 1-antagonist) and ICI 118-551 (beta 2-antagonist) were utilized to differentiate both subclasses unequivocally. beta-Adrenoceptors in rat kidney were found to be almost exclusively beta 1. They were located mainly on glomeruli and to a lesser extent on the straight part of the distal tubules and on the cortical portion of the collecting ducts. Some beta 2-adrenoceptors were localized around the corticomedullary junction. Grain localization in the auto-radiograms was absent in the inner medulla and papilla. Glomeruli and distal tubules of the guinea-pig kidney also possess only beta 1-adrenoceptors, but, in contrast to the rat, extremely high concentrations of beta 2-adrenoceptors were associated with the straight part of the proximal tubules in the cortex and possibly with the cortical portion of the collecting duct. Labelling was not detected on the proximal convoluted tubule in either species. PMID- 2984589 TI - Forskolin inhibits potassium-evoked release of vasopressin from rat neurohypophyses. AB - The effects of forskolin, an activator of adenylate cyclase, on the release of vasopressin from isolated rat neurointermediate lobes during a 10 min period of potassium stimulation were investigated. Forskolin was added 5 min after the onset of potassium stimulation. A concentration-dependent reduction in the amount of hormone released during the remaining 5 min period of stimulation was observed. The results suggest that an increase in cyclic AMP following depolarisation inhibits hormone release. This contrasts with several other reports showing that an increase in cyclic AMP prior to stimulation enhances release. We therefore propose that cyclic AMP plays a dual role in stimulus secretion coupling in the neurohypophysis. PMID- 2984590 TI - [Bone scintigraphy in oncology, when is it appropriate?]. PMID- 2984592 TI - The diagnostic use of monoclonal antibodies against small cell lung carcinoma cells. PMID- 2984591 TI - Regulatory role of macrophages in tumor allograft development. AB - In the study of the role of macrophages in antitumor immune response the model of Sa I (H-2a) allograft development in B10 mice (H-2b) treated with xenogenous antithymocyte serum (ATS) was used. In the treated recipients tumor growth was enhanced as compared to untreated mice, subsequently in some of the mice the tumor permanently regressed while in the rest the temporary regression was followed by a secondary progressive growth. To determine the role of macrophages at different periods of tumor development, both silica (which is known to damage the function of macrophages) and double-stranded RNA (dsRNA; which, in turn, stimulated their function), were used. The administration of silica on the day of tumor cell transplantation slightly promoted tumor regression, whereas silica administered on days 7 or 14 after the cell transfer enhanced the secondary growth of the tumor. On the other hand, administration of dsRNA on the day of tumor transplantation promoted tumor growth, whereas, when administered on days 7 or 14, it enhanced the regression of the tumor. An administration of both substances at later periods had no effect on the growth of the tumor. From these results it can be assumed that macrophages at the time of induction of antitumor immune responses participate in the events enhancing the subsequent growth of the tumor allograft, whereas at later times they have an antitumor activity. PMID- 2984593 TI - The significance of the absence of 131I-Hippuran uptake by a kidney graft. AB - Anuria is a frequent complication during the immediate post-transplantation period in kidney transplant recipients. Radioisotopic studies with 131I-Hippuran and 99mTc-pertechnetate allow correct differentiation between viable and non viable grafts. PMID- 2984594 TI - Evidence for the participation of disulfide and sulfhydril groups in the specific binding of [3H]prazosin in cerebral cortex. AB - The tritiated alpha 1 antagonist prazosin [3H]PRZ binds specifically and with high affinity to postsynaptic adrenoceptors in membrane preparations from cerebral cortex. Since adrenoceptors are of protein nature, it was of interest to investigate the possible role of disulfide (--SS--) and sulfhydril (--SH) groups in the binding of [3H]PRZ. Pretreatment of the membranes with the disulfide and sulfhydryl reactives DL-Dithiothreitol, L-Dithiothreitol, Dithioerythritol or 5',5'-Dithiobis-(2-nitrobenzoic acid) (DTNB), alone or in combination with the alkylating agent N-Methylmaleimide (NMM), decreased specific [3H]PRZ binding, with minor changes in the non-specific counts. Saturation experiments revealed that all these reagents reduced the affinity of the binding site for [3H]PRZ, as judged by the Kd 25 degrees C, but only the alkylating agent NMM and the oxydizing reagent DTNB produced in addition to the increase in Kd, a decrease of the maximum binding capacity (Bmax). The present results provide evidence for a participation of --SS-- and/or --SH groups in the recognition site of the alpha 1 adrenoceptor of cerebral cortex. PMID- 2984595 TI - Postnatal ontogeny of uridine kinase in the cerebellum, hypothalamus, and cerebral cortex of the rat. AB - Postnatal developmental patterns of uridine kinase were determined in crude subcellular fractions of the rat cerebellum, hypothalamus and cerebral cortex at ages 3 through 60 days. The highest specific activity and predominant distribution of enzyme was in the 105,000 g supernatant of the 3 brain regions. Enzyme activity in hypothalamus and cerebral cortex was maximum at 3 days and decreased with age; in cerebellum it increased through 13 days and decreased thereafter. Thus, the pattern of activity in hypothalamus and cerebral cortex paralleled changes in DNA and RNA synthesis through age 60 days; in cerebellum, it more closely approximated changes in DNA synthesis during early development. Changes in Km with aging suggest that the brain regions contain more than one form of enzyme. The highest particulate activity was in the microsomal fraction of the cerebellum and hypothalamus at all ages and in the cortex at 35 and 60 days. Relative specific activity for microsomal fractions of the brain regions at 60 days indicate a concentration of the enzyme which may be relevant in the maintenance of RNA activity in adult brain. PMID- 2984596 TI - [Cerebral blood flow measured by Xenon enhanced CT in brain tumors]. AB - In the management of malignant brain tumors, it is important to know the extent and viability of tumors. However, an ordinary CT scan with iodine enhancement has only a limited ability to distinguish the tumor from surrounding normal tissue. Since the blood flow in tumor tissue was found to be relatively high in a previous experimental report, we have investigated the blood flow in a tumor and the surrounding brain. The Xenon enhanced CT method has several advantages over the conventional isotope method and enables us to evaluate rCBF with the same resolving power as with the CT scan. We evaluated rCBF in 15 brain tumor cases and obtained the following results. Mean rCBF value of the tumor is a little lower than that of gray matter and higher than that of surrounding edema. Our Xe CT method enables us to distinguish the demarcation between the tumor area and the surrounding edematous area and offers useful information for determining the extent of resection in surgery. Mean lambda value of the tumor which is not obtainable in vivo by radionuclide scanning, was 1.02 +/- 0.06 for gliomas and 0.72 +/- 0.09 for metastatic tumors. rCBF value and lambda value are important elements to know the uptake rate of anticancer drugs into the brain tumors. And to evaluate these value in each brain tumor is useful in the selection of chemotherapeutic agents. PMID- 2984597 TI - [Spinal epidural metastasis of glioblastoma multiforme: a case report]. AB - It is generally accepted that the metastases of intracranial glioma to extracranial location are rare. In such a case the minimal criteria proposed originally by Weiss should usually be satisfied if a report is to be considered as an acceptable case of metastasizing central nervous system glioma outside the central nervous system. We report a case of glioblastoma multiforme, fulfilling Weiss' criteria, metastasizing the spinal epidural space. The patient was a 32 year-old male, who underwent craniotomy and subtotal removal of a glioblastoma multiforme in the left parietooccipital area. He was additionally treated with irradiation and chemotherapeutic agents. Twelve months after the craniotomy, he was admitted again to our clinic because of sudden onset of severe lumbago, paraplegia and urinary disturbance. Diagnosis of a spinal epidural tumor was made and laminectomy (Th10-L1) was performed. At operation, an epidural mass was found, however no invasion to the spinal cord or dura was noted. Histological diagnosis of the tumor was glioblastoma multiforme. Although he was treated with radiation, pulmonary metastasis was manifested one month later, and the condition of the patient deteriorated. He died 21 months after the first operation and 8 months after the second operation. Even at the terminal stage, his consciousness was clear without any sign for recurrence of intracranial tumor. The general autopsy was done and multiple metastatic lesions of glioblastoma multiforme in paratracheal and paraaortic lymph node, left pleura, both lungs and spinal cord were observed. The present case suggests that the surgical intervention, irradiation, and chemotherapy may contribute to extracranial metastasis of a glioblastoma. PMID- 2984598 TI - Effect of (D-Met2, Pro5)enkephalinamide, a highly potent opiate agonist, on the drinking behaviour of rats. AB - The highly potent opiate agonist, (D-Met2, Pro5)enkephalinamide, markedly influenced drinking behaviour of the rat. It inhibited the drinking caused by water deprivation, the drinking response to intracerebroventricular application of angiotensin-II, as well as the drinking evoked by systemic administration of isoprenaline. The compound investigated was effective not only upon intracerebroventricular injection but also upon systemic application. The LD50 values were low; 0.42 mg/kg after intraperitoneal administration and about 1.1 micrograms after intracerebroventricular injection. The findings not only support the view that opioid receptors are involved in one way or other in the regulation of fluid intake of the rat, but indicate also an interesting peptide-peptide interaction within the brain. PMID- 2984599 TI - Postoperative prophylactic anticonvulsant therapy in cerebral gliomas. AB - A retrospective study was performed to evaluate the efficacy of prophylactic anticonvulsants in preventing seizures in 68 patients with supratentorial astrocytomas who had been treated with operation and irradiation and who had no previous history of convulsions. Thirty-three patients received prophylactic anticonvulsants and 38 patients did not. The incidence of all types of seizures (generalized convulsions or partial) was lower in patients receiving anticonvulsants. No seizures with an impairment of consciousness occurred in the patients with documented therapeutic anticonvulsant blood levels. The overall incidence of seizures was 39% in untreated patients and 21% in treated patients. The incidence of major seizures including tonic/clonic or partial complex seizures with impairment of consciousness was zero in patients with therapeutic anticonvulsant levels and 18% in untreated patients. Regarding the overall incidence of seizures in both groups, there tend to be fewer seizures in older patients, females, patients with a higher grade of malignancy, and patients who had a more radical resection of the tumor. This study suggests that seizures are a frequent occurrence after operation and irradiation for supratentorial glioma and that anticonvulsants may be effective in reducing the incidence of those seizures. PMID- 2984600 TI - Neuronal antinuclear antibody in sensory neuronopathy from lung cancer. AB - We found an antinuclear antibody highly restricted to nuclei of neurons in two patients with subacute sensory neuronopathy complicating oat cell carcinoma of the lung. Serum was tested by indirect immunofluorescence and immunoperoxidase staining. At low concentrations of antibody, only the nuclei of the neurons were stained. At high concentrations, there was also staining of the nuclei of glial cells and fetal nonneural tissues. The cytoplasm of most neurons was stained with the immunoperoxidase method. PMID- 2984602 TI - Relapsing ophthalmoparesis-sensory neuropathy syndrome. AB - We studied two patients with recurrent sensory neuropathy and weakness of extraocular muscles. There was electrophysiologic evidence of multifocal demyelination of sensory nerves, with relative sparing of somatic motor nerves. Sural nerve biopsy in one patient showed segmental demyelination. We believe that these patients had an unusual form of inflammatory polyneuropathy--possibly a relapsing variant of the Miller-Fisher syndrome of acute idiopathic polyneuritis. PMID- 2984601 TI - Cytomegalovirus meningoencephalitis in a homosexual man with Kaposi's sarcoma: isolation of CMV from CSF cells. AB - A 36-year-old homosexual man developed a lymphocytic meningoencephalitis, lymphadenopathy, decreased helper/suppressor ratio in peripheral blood, and Kaposi's sarcoma. After repetitive evaluations for bacterial and fungal agents were negative, viral cultures were positive for cytomegalovirus. Virus was isolated only from CSF cells obtained from a large volume of fresh CSF. We present this case to document the association of CMV meningoencephalitis and acquired immunodeficiency syndrome (AIDS), and to emphasize that aggressive attempts to isolate CMV from CSF cells may be warranted in undiagnosed meningoencephalitis in patients at risk for AIDS. PMID- 2984603 TI - Waldenstrom's macroglobulinemia and peripheral neuropathy: deposition of M component and kappa light chain in the endoneurium. AB - Some cases of peripheral neuropathy associated with benign IgM monoclonal gammopathy, or Waldenstrom's macroglobulinemia, are probably of autoimmune origin; in some cases, anti-IgM serum reacts with the myelin sheaths of peripheral nerves. However, mechanisms may differ in other cases. In one case of neuropathy with macroglobulinemia, we found deposits of IgM immunoglobulin in the endoneurium. On ultrastructural examination, the deposits had erased the basement membrane of some nerve fibers that showed damage of both myelin and axon. PMID- 2984604 TI - Synovial sarcoma, a nasopharyngeal location. PMID- 2984605 TI - [Long-term treatment of secondary hyperparathyroidism in the chronic hemodialysis patient with 1,25-(OH)2D3]. PMID- 2984606 TI - Increased binding of calcium in the hippocampal slice during long-term potentiation. AB - Long-term potentiation (LTP) of CA1 pyramidal neurons was induced by tetanic stimulation in the stratum radiatum of hippocampal slices from guinea pigs. Unstimulated and stimulated slices were treated using a histochemical procedure enabling the electron microscopic (EM) visualization of Ca binding sites. Electron-dense, Ca-containing deposits were found in low numbers in unstimulated slices on pre- and postsynaptic sites. In the stratum radiatum of tetanized slices the overall number of deposits as well as the number of deposits in dendrites was clearly increased. The results support the hypothesis that Ca dependent postsynaptic mechanisms are important for the generation of LTP. PMID- 2984607 TI - Functional alteration of opioid receptor subtypes in the mice exhibited conditioned suppression in motility. AB - Mice exhibit a marked suppression of motility (conditioned suppression) when placed in the same environment in which they had previously received the electric footshock. The present study was designed to investigate the functional change of opioid receptor subtypes in the conditioned suppression group using an opioid binding assay technique. In the synaptic membrane of the conditioned suppression group, the binding capacities of [3H]naloxone at high and low affinity binding sites and of [3H]phencyclidine at high affinity binding site were significantly increased compared to those of the control group. On the other hand, the binding capacity of [3H]ethylketocyclazocine at both affinity binding sites in the conditioned suppression group was not changed. These results suggest that the binding function of different opioid receptor subtypes may be altered differently by stress. PMID- 2984609 TI - Tonic depolarization of excitatory nerve terminals in crayfish muscle by high concentrations of extracellular potassium. AB - At voltage-clamped fibres of the claw opener muscle of small crayfish, spontaneous quantal release of excitatory transmitter elicited by raising extracellular K+ to 100 mM was investigated. On application of the high K+ concentration, the rates of quantal release increased to n = 10,000-25,000 quanta/s within 10 s, and thereafter declined exponentially, either with a single (tau congruent to 15-40 s) or with two (tau 1 congruent to 15-40 s, tau 2 greater than 70 s) time constants. The total number of quanta released per trial ranged from s = 200,000 to 800,000 quanta. The results were derived by means of the fluctuation analysis technique. PMID- 2984608 TI - Evidence that the anticonflict effect of midazolam in amygdala is mediated by the specific benzodiazepine receptors. AB - The benzodiazepine midazolam produced an anticonflict effect in rats measured in a water lick paradigm following local injection into the basolateral and lateral complex of the amygdala. This effect of midazolam seems to involve specific benzodiazepine receptors, since the systemic injection of benzodiazepine antagonists Ro 15-1788, ZK 93426, FG 7142 and CGS 8216 produced strong antagonism of the effect of midazolam in doses not affecting the non-punished drinking behaviour. PMID- 2984610 TI - Localization of aspartate aminotransferase and cytochrome oxidase in the cat retina. AB - Using immunohistochemical techniques, we demonstrate aspartate aminotransferase (AAT)-like immunoreactivity in cone pedicles and ganglion cells of the cat retina. An identical pattern was seen when we stained for cytochrome oxidase activity, a marker for neurons which have a high metabolic activity. Tetrodotoxin selectively blocked the cytochrome oxidase labeling of ganglion cells. AAT is a key enzyme in the metabolism of aspartate and glutamate and has been proposed as a marker for neurons which use aspartate/glutamate as a neurotransmitter. Due to the close correlation between AAT-like immunoreactivity and cytochrome oxidase activity, we suggest that, at least in the retina, AAT-like immunoreactivity in fact labels cells which have a high metabolic activity. PMID- 2984611 TI - Bartsocas-Papas syndrome. A case report. PMID- 2984612 TI - The transfer of information into cells. AB - Our understanding of cellular events involved in the transfer of information between and within cells has increased dramatically in the last decade. The application of this information to clinical medical research is just beginning. The relevance of these processes to the action of hormones and drugs and developmental signals makes it inevitable that increased understanding will lead to rational and innovative therapeutic approaches. PMID- 2984613 TI - Primary nonepithelial cancers of the ovary. PMID- 2984614 TI - Colposcopic findings associated with human papillomavirus infection of the vagina and the cervix. PMID- 2984615 TI - A review of lower genital intraepithelial neoplasia and the use of topical 5 fluorouracil. PMID- 2984616 TI - Amenorrhea and endometrial atrophy with continuous oral estrogen and progestogen therapy in postmenopausal women. AB - An oral regimen of continuous conjugated equine estrogens (Premarin 0.625 or 1.25 mg daily) and low-dose progestogen (Norethisterone 0.35 to 2.1 mg daily) have been used to treat 95 nonhysterectomized postmenopausal women for up to 2.5 years. This method of hormone replacements was undertaken in an attempt to avoid the withdrawal bleeding and progestogenic side effects associated with conventional cyclical therapy with estrogen and progestogen, while simultaneously protecting the endometrium from estrogenic over-stimulation. With the lower dose of estrogen, amenorrhea was achieved immediately in 30 of 46 patients (65%), and after adjustments to the dose of the progestogen in all ten patients observed for at least one year (maximum 2.5 years). With the higher dose of estrogen, irregular spotting during the first three months resulted in the cessation of treatment by six of the 49 patients (12%), but 23 (47%) women had no bleeding during that time; by 15 months, all 13 patients who had remained in treatment had become amenorrheic (maximum 2.25 years). Endometrial biopsy specimens after six months of combined treatment in 56% of patients revealed atrophic histology regardless of the dose of the estrogen. PMID- 2984617 TI - A comparative clinical trial of the contraceptive sponge and Neo Sampoon tablets. AB - Neo Sampoon, a foaming vaginal tablet containing 60 mg of the spermicide menfegol, and the Collatex sponge (now marketed in the United States as the Today sponge), a dome-shaped polyurethane device that contains 1 g of nonoxynol-9, were compared in terms of effectiveness, safety, and acceptability. Both methods were new to the Maribor General Hospital, Yugoslavia, where the trial was conducted among 450 volunteers randomly assigned to one of the two methods. At 12 months, the life-table pregnancy rate per 100 women for the Neo Sampoon group was 12.8, compared with a rate of 10.4 among the sponge users (P greater than .10). After pregnancy, the second most frequent reason for termination was discomfort, with a 12-month termination rate due to this cause of 6.9 per 100 women in the Neo Sampoon group and 6.2 in the sponge group. Although fewer than a quarter of the volunteers had any experience with barrier methods before this trial, the life table continuation rate was high in both groups, with more than 70% using their assigned method for the full 12 months. Also, upon conclusion of the study, 41% of the volunteers chose another barrier contraceptive method. Although the effectiveness of the sponge and Neo Sampoon is not comparable to that of the pill or IUD, both vaginal methods appear to be safe and acceptable additions to the range of contraceptive choices. PMID- 2984618 TI - Clinical and laboratory investigation of a virilized woman with placental-site trophoblastic tumor. AB - The novel association of virilization and placental-site trophoblastic tumor, an uncommon form of gestational trophoblastic disease (formerly called "trophoblastic pseudotumor"), is described in a 32-year old woman. Multiple agent chemotherapy lowered serum concentrations of chorionic gonadotropin (hCG) (8.7 to 2.1 ng/mL), pregnancy-specific beta 1-glycoprotein (32 to 3.9 ng/mL), and testosterone (400 to 74 ng/dL). Subsequent hysterectomy revealed a 2-cm tumor nodule within the myometrium, and one week postoperatively serum concentrations of hCG (0.5 ng/mL), pregnancy-specific beta 1-glycoprotein (1.3 ng/mL), and testosterone (20 ng/dL) had all returned to normal. Percutaneous catheterization of the ovarian veins before chemotherapy demonstrated a 50-fold elevation of ovarian vein testosterone concentrations compared with normal women, whereas ovarian vein 17 beta-estradiol concentrations were only twofold higher than normal. Direct sampling of the uterine veins at the time of hysterectomy documented testosterone and estradiol production, presumably by the placental site trophoblastic tumor. Ovarian vein testosterone concentrations at this time were only fourfold above normal, probably the result of falling serum concentrations of hCG. Wedge biopsy of the ovaries disclosed minimal histologic changes (thecal cell luteinization, focal thecosis) compatible with hCG stimulation. Failure of the placental-site trophoblastic tumor to produce large amounts of estrogen, in contrast to normal pregnancy and hydatidiform mole, resulted in marked androgen/estrogen imbalance, high circulating concentrations of free testosterone, and virilization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984619 TI - Microorganisms in semen used for artificial insemination. AB - The effect of freezing semen in a cryopreservative media consisting of egg yolk glycerol with or without erythromycin was tested for its effect on the viability of microorganisms present in donor semen and on sexually transmitted pathogens seeded into semen. All donor semen contained two or three species of microorganisms that could be considered skin flora. Five of ten donor semen specimens contained Ureaplasma urealyticum that was not affected by either freezing or antimicrobial treatment. Some strains of Neisseria gonorrhoeae, when seeded into semen, survived all conditions except freezing in egg yolk glycerol containing erythromycin. Chlamydia trachomatis was erradicated when erythromycin was present in the cryopreservative. There was no detectable effect of any treatment tested on the survival of herpes simplex virus. PMID- 2984620 TI - Primary sarcoma of the adult vagina: a clinicopathologic study. AB - Combination chemotherapy has dramatically improved the results of treatment for embryonal rhabdomyosarcoma of the vagina in children, but little attention has been directed toward vaginal sarcomas in adults. This report of 17 cases of primary sarcoma of the adult vagina includes ten leiomyosarcomas, four mixed mesodermal tumors, one undifferentiated sarcoma, one stromal sarcoma, and one neurofibrosarcoma. The leiomyosarcomas, mixed mesodermal tumors, and the stromal sarcoma were morphologically similar to their uterine counterparts. Thirty-five percent of the patients had received pelvic irradiation for carcinoma of the cervix. Three of four patients treated by pelvic exenteration are alive and disease free after 84 to 161 months of observation. All 13 patients who received other forms of primary therapy died of recurrence. The pelvis was the first site of recurrence in all treatment failures and the only site of failure in 50%. PMID- 2984621 TI - Malignant mixed mesodermal tumor of the ovary: a report of 22 cases. AB - Clinicopathologic data are presented for 22 cases of malignant mixed mesodermal tumors of the ovary of both homologous and heterologous types. The results substantiate previous reports of low parity in the almost exclusively postmenopausal women with this tumor. Symptoms were the same as for ovarian malignancy in general. More than half the patients presented with stage III or IV disease, according to the International Federation of Gynecology and Obstetrics (FIGO) classification. Only four (19%) of 21 patients were alive at 18 months and had survived from three to more than 13 years. Their survival was associated strictly with stage I or II disease and with the purely homologous stromal pattern. Two tumors were contiguous with remnants of ovarian endometriosis. Differentiated malignant epithelium was most often of the serous/endometrioid type, frequently with squamous zones, and was of the mucinous type in only one case. Whereas spindle cell stroma was present in many tumors of both stromal types, atypical cartilage was the predominant heterologous element. PMID- 2984622 TI - Studies on cataractogenesis in humans and in rats with alloxan-induced diabetes. I. Cation transport and sodium-potassium-dependent ATPase. AB - Changes in the cation balance cause hydration and initiate the process of lens opacification. Such alterations were studied in human cataractous lenses and during the development of alloxan-induced diabetic cataract in rats by biochemical and histochemical techniques. The development of alloxan-induced cataract in rats was examined in vivo which showed cortical opacities beginning after 32 days. These opacities did progress to maturity after 64 days and finally the lenses were completely opacified after 96 days of alloxan treatment. The histochemical localization of sodium-potassium-activated adenosine triphosphatase using three different methods provided information on the possible role of this enzyme in normal and cataractous lenses. In human cataractous lenses, sodium potassium adenosine triphosphatase activity was found to be considerably decreased, whereas no activity of this enzyme was localized in human diabetic cataractous lenses. An animal model provided evidence that an apparent decrease of sodium-potassium adenosine triphosphatase may be involved in the initiation of alloxan-induced diabetic cataract in rats. PMID- 2984623 TI - Studies on cataractogenesis in humans and in rats with alloxan-induced diabetes. II. Histochemical evaluation of lenticular enzymes. AB - The effect of cataractogenesis on the behavior of some enzymes involved in glucose metabolism was examined histochemically both in human lenses and in rat lenses from rats with alloxan-induced diabetes. Several modifications in the currently available techniques were made in order to localize glucose-6-phosphate dehydrogenase, aldose reductase, sorbitol dehydrogenase, hexokinase and ketohexokinase in ocular lens. Human cataractous lenses showed a precipitous drop in glucose-6-phosphate dehydrogenase activity, whereas the lenticular tissues of alloxan-treated rats showed a gradual decrease of this enzyme with the prolongation of diabetes. Aldose reductase activity increased in hypermature and senile diabetic cataracts, whereas sorbitol dehydrogenase activity decreased in these lenses. Similarly, in alloxan-diabetic rat lenses the activity of aldose reductase increased while that of sorbitol dehydrogenase decreased with the prolongation of diabetes. Attempts were made to localize hexokinase and ketohexokinase in ocular lens. PMID- 2984625 TI - Current results of proton beam irradiation of uveal melanomas. AB - Proton beam irradiation has been used for the treatment of 241 uveal melanomas over the past 7 1/2 years. Twelve melanomas (5%) were small, 99 (41%) medium, 103 (43%) large and 27 (1%) extra-large melanomas. The mean length of follow-up was 21 months and the median 15 months. Ninety-four percent of the treated lesions with a follow-up more than two years and 65% of tumors with shorter follow-up showed regression. The most recent visual acuity was 20/40 or better in 47% and 20/100 or better in 66%. Ten eyes were enucleated because of complications (9) or continued tumor growth (1). Thirteen patients developed metastases from 4 to 50 months of treatment. Our data indicate that proton irradiation can be used to treat melanomas of various sizes and in a variety of locations, and preliminary results suggest that proton therapy has no deleterious effect on the likelihood of the development of metastases. PMID- 2984624 TI - Adenoid cystic carcinoma of the orbit diagnosed by means of aspirative cytology. AB - A case of intraorbital adenocystic carcinoma, which was diagnosed preoperatively by means of fine needle aspiration, is described and discussed. PMID- 2984626 TI - Hydroxylapatite reconstruction of alveolar ridge deficiency with an open mucosal flap technique. AB - A preliminary report on an open surgical technique for mandibular augmentation with hydroxylapatite has been presented. Twelve patients were evaluated from 2 to 20 months postoperatively. Advantages are thought to include better ridge form, maintenance of vestibular depth, and visualization of the mental nerve. In addition, splints usually are not necessary. Disadvantages include increased surgical time, possible lower lip inversion, and soft-tissue characteristics that may require secondary revisions. PMID- 2984628 TI - Primary giant cell carcinoma of the larynx. A clinico-pathological study of four cases. AB - The clinical and pathological findings in 4 cases of primary giant cell carcinoma of the larynx are reported. This variant of large cell carcinoma resembles the well-defined giant cell carcinoma of the lung. It has to be distinguished from other types of carcinoma of the larynx. This can be difficult on small biopsy specimens and the definite diagnosis may only be established after the detailed examination of the excised tumour. The highly malignant behavior of the laryngeal type was noted. To our knowledge this is only the second report on giant cell carcinoma of the larynx. PMID- 2984627 TI - Combined effect of herpes simplex virus and tobacco on the histopathologic changes in lips of mice. AB - In the present study we have examined the combined effect of HSV-1 inoculation and tobacco application (snuff water extract or smoking tar condensate) on the histopathologic changes of mouse labial mucosa. Two months' exposure to tobacco or HSV-1 inoculation alone did not induce dysplasia in the epithelium of labial mucosa, while HSV-1 inoculation combined with snuff water extract or smoking tar condensate produced epithelial dysplasia and other histomorphologic changes (that is, hyperkeratosis, increased granular cell layer thickness, acanthosis, and increased inflammatory cell infiltration in a significant number of animals). This result indicates that HSV-1 and tobacco could possibly act synergistically in the development of precancerous oral lesions and oral cancer. PMID- 2984630 TI - Granular cell tumor of facial nerve diagnosed at surgery for idiopathic facial nerve paralysis. PMID- 2984629 TI - [Polymyxin B sulfate-induced non-specific bronchial provocation in bronchial asthma and chronic bronchitis]. PMID- 2984631 TI - Diagnosis of hyperparathyroidism. AB - Availability of immunoassays for specific regions of the parathyroid hormone (PTH) molecule allows discrimination with a high level of surety between primary hyperparathyroidism and tumoral hypercalcemic states associated with circulating PTH-like substances. Assay for intact, N-terminal PTH currently has the highest discriminant function. Prostaglandin-dependent and osteoclast-activating factor mediated hypercalcemic states associated with neoplasia have suppressed serum PTH levels. PTH-like substances are detected by immunoassays, but in the intact, N terminal system they are seen as normal-range or low values. The frequency with which any tumor produces only authentic PTH is very low. The serum chloride:phosphate ratio has limited clinical utility in distinguishing tumoral hypercalcemia from hyperparathyroid hypercalcemia, and measurements of nephrogenous cyclic AMP do not distinguish between the effects of circulating authentic PTH and PTH-like substances elaborated by tumors. Additional measures that, in the future, may help to distinguish between parathyroid and tumoral hypercalcemias include quantitative bone biopsy histomorphometry and in vitro bioassays for PTH activity in the separate plasma fractions, obtained by gel filtration, in which PTH and PTH-like substances are found. PMID- 2984633 TI - The effect of laminectomy on spinal cord blood flow, energy metabolism and ATPase activity. PMID- 2984632 TI - Evaluation of a low dose administration of aspirin, dipyridamol and steroid. Therapeutic effects on motor function and protective effects on Na+-K+-activated ATPase activity against lipid peroxidation in an experimental model of spinal cord injury. PMID- 2984634 TI - [Effect of reflexotherapy on the function of the hypophyseal-adrenal system]. PMID- 2984635 TI - Glycerol kinase deficiency inhibits glycerol utilization in phosphoglyceride and triacylglycerol biosynthesis. AB - Glycerol 3-phosphate is an initial metabolite in the biosynthesis of phosphoglycerides and triacylglycerols. Both glycerol and glucose are precursors of glycerol 3-phosphate. Cultured skin fibroblasts from patients with glycerol kinase deficiency utilized glucose, but not glycerol in the biosynthesis of phosphoglycerides and triacylglycerols. Phosphoglyceride and triacylglycerol biosynthesis in glycerol kinase deficiency fibroblasts is not diminished by the inability to use glycerol as a precursor of glycerol 3-phosphate. PMID- 2984636 TI - Long-term protective immunity of recipients of the OKA strain of live varicella vaccine. AB - In spite of close contacts with patients who had varicella, 101 of 106 (95%) healthy and sick children (142 of 147 (97%) exposures of these children) who had received the OKA strain of live varicella vaccine 7 to 10 years earlier were protected against the disease completely. Among them, 37 of 38 (97%) vaccine recipients who received immunologic testing had varicella-zoster virus (VZV) antibodies tested by fluorescent antibody to membrane antigen method with a geometric mean titer of 1:9.3, and 37 of the 38 (97%) showed positive skin reaction to varicella-zoster virus antigen with erythema (mean diameter 13.4 mm). These findings were compared with those for 29 children who had contracted typical varicella 7 to 10 years earlier, whose seropositive rate was 100% with a geometric mean titer of 1:10.5, and 97% of whom (28/29) had positive skin reaction with mean diameter of 12.9 mm. These results indicate that the vaccine induced protective immunity persists for approximately one decade and is almost equal to the long-term immunity following natural infection. PMID- 2984637 TI - Neurologic manifestations of Schoenlein-Henoch purpura: report of three cases and review of the literature. AB - Three patients developed prominent neurologic symptoms and signs associated with Schoenlein-Henoch purpura. A 7 1/2-year-old boy was seen with status epilepticus after a 2-week history of generalized headaches, irritability, and intermittent colicky abdominal pain. A left hemiparesis and a left homonymous hemianopia with a right gaze preference that were present on initial examinations gradually resolved, but a mild left arm paresis persisted. Cutaneous, renal, and joint involvement followed initial CNS manifestations. The second patient, a 7-year-old girl, had a complex partial seizure with secondary generalization and a postictal hemiparesis seven days after presentation with classic signs of Schoenlein-Henoch purpura. Behavioral changes were noted during the acute phase of the illness. The third patient, a 13-year-old boy, developed signs of a left brachial plexopathy and transient weakness of his right leg during a complicated course of Schoenlein Henoch purpura. Review of the world literature indicates that headaches and mental status changes are the most frequent neurologic complications of Schoenlein-Henoch purpura, followed by seizures, focal neurologic deficits, mononeuropathies, and polyradiculoneuropathies. The vasculitis of Schoenlein Henoch purpura can involve the nervous system and may add significantly to the morbidity of the illness. PMID- 2984638 TI - Halopemide, a new psychotropic agent. Cerebral distribution and receptor interactions. AB - Halopemide is a new psychotropic agent, a structural analogue of the neuroleptics of the butyrophenone type but with different pharmacological and clinical properties. Preliminary clinical findings indicate that halopemide lacks the ability to induce parkinsonism and may be an effective drug in the treatment of psychosis characterized by autism, emotional withdrawal or apathy. Its pharmacological effects at a molecular level in comparison to structurally related neuroleptics and putative metabolites are reviewed. PMID- 2984641 TI - Nutrition. 4. High fibre diets. PMID- 2984639 TI - Short term effect of low doses of tri-iodothyronine on proximal tubular membrane Na-K-ATPase and potassium permeability in thyroidectomized rats. AB - Tri-iodothyronine (T3), even when administered for short time and at low doses, induces a large increase in the isotonic fluid reabsorption (Jv) in proximal tubules of thyroidectomized rats (TX). In order to investigate the role of the Na K-ATPase in this process, we measured the Na-K-ATPase activity in early proximal convoluted tubules (S1) and proximal straight tubules (S2) microdissected from TX rats and rats treated with low doses of T3 (10 micrograms/kg body wt), either for 3 days (TX + 3T3) or for 7 days (TX + 7T3). In both segments no changes in Na-K ATPase activity were found in TX + 3T3 rats versus TX rats, while an increase was registered in TX + 7T3 rats. Using micropuncture techniques, Jv measured on the same tubular segments increased by 68% in TX + 3T3 rats versus TX. Thus, no correlation between Jv and Na-K-ATPase activity measured in vitro could be detected after short term treatment of TX rats with T3. Na-K-ATPase activity in vivo is also regulated by the potassium permeability of the membrane, which might be altered by tri-iodothyronine. This hypothesis was tested by perfusing intraluminally and peritubularly proximal tubules of TX rats with the K ionophore, valinomycin (1 microgram/ml). In the dual perfusion experiments valinomycin elicited 40% of the action induced on Jv by 3 days treatment with T3. On the other hand, no further increase in Jv was recorded when valinomycin was applied in TX rats pretreated with T3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984640 TI - Adenosine-induced depression of synaptic transmission in the isolated olfactory cortex: receptor identification. AB - We have investigated the type of purine receptor in the guinea-pig olfactory cortex, using pial surfaces slices maintained in vitro. Adenosine (0.1 to 100 mumol/l) bath applied in the presence of the uptake inhibitor nitrobenzylthioinosine, depressed the evoked potentials in a dose related fashion. Synthetic and uptake resistant adenosine analogues had the same effect as adenosine and the order of potency of these was: 5'-N-ethylcarboxamide adenosine greater than L-N6-phenylisopropyl adenosine (L-PIA) = N6 cyclohexyladenosine = 2-chloroadenosine greater than adenosine greater than D-N6 phenylisopropyladenosine (D-PIA). The D-stereoisomer of PIA was 45 times less potent than L-PIA. The methylxanthine compounds 8-phenyltheophylline (3 mumol/l) and 3-isobutyl-1-methylxanthine (50 mumol/l) antagonised the depression produced by L-PIA. Rolipram, a phosphodiesterase inhibitor, in concentrations up to 100 mumol/l had no effect on the evoked potentials or on adenosine action. Forskolin, a cAMP stimulant, slightly increased the amplitude of the evoked potential, and partly reversed the depressant effect of adenosine. Noradrenaline had no effect either alone or in the presence of adenosine. The results of these experiments indicate the existence of A1 subtype adenosine receptors in the guinea pig olfactory cortex probably linked to a depression of intracellular cAMP. PMID- 2984643 TI - [Pancreatic insulinoma in a patient with Hodgkin's disease]. PMID- 2984642 TI - Epidemiology of cytomegalovirus infections in young children: day care vs. home care. AB - Infection rates with cytomegalovirus among children in three day care centers were compared to that found in a group of children cared for in the home who were from a similar socioeconomic background. Rates of viral excretion for Day Care Centers 1, 2 and 3 were 41% (28 of 68), 26% (15 of 58) and 55% (34 of 62), respectively, with a combined rate of infection for all children in day care of 41% (77 of 188). In contrast 15% (10 of 66) of children in home care were seropositive to cytomegalovirus, and only 2 of 25 (8%) were shedding virus (P less than 0.001). Although the median age of the group in home care was slightly lower, the two groups of children were similar in sex, race, breast-feeding and parental ages and educational background. The only other notable difference between the children in day care and those in home care was the environment in which they received care for almost 40 hours every week. Rates of infection varied also among the age groups within each center, with the highest rate within each center occurring in children 25 to 36 months old. Day care for young children is likely to be associated with high rates of infection with cytomegalovirus. The mechanisms of transmission that are responsible for these higher than expected rates of infection remain to be defined. PMID- 2984644 TI - [Effect of polymyxin B on the bronchi and peripheral blood basophils (in vitro) in patients with atopic asthma]. PMID- 2984645 TI - Cervical dysplasia: cytology class III D and CIN I-II. AB - Some aspects of class III D-smears (suggesting slight to moderate dysplasia) have been investigated. Results of a cytological follow-up in 481 patients and of the histological findings immediately or during the follow-up in 142 patients are presented. In 112 patients a triple cervical smear approach has been used to determine the rate of false negative smears and errors of sampling. This approach includes the preparation of three slides with cellular material obtained separately from the anterior cervical lip, the posterior lip and from the endocervical canal, using three cotton applicators. To compare the results of cytology with the grade of CIN in different areas of the conization specimens triple cervical smears have been taken preoperatively. Finally 28 patients with a conization following a class III D-smear have been investigated for HPV-induced cell changes which were found in six cases. PMID- 2984646 TI - What's new in colon carcinogenesis? AB - Among precancerous conditions in the colon and rectum, adenomas are the most important, paradigmatically so in adenomatosis coli, a genetic predisposition of autosomal dominant type. Statistical relevance is also attributed to long-term ulcerative colitis, Morbus Crohn, and Gardner's syndrome. Dietary factors seem to play an essential role. Highly developed countries in Western Europe and North America whose prevalence rates of colorectal carcinoma are highest, also have a particularly high daily intake of fat and meat. The proportion of fibers in a diet shows an inverted correlation with the prevalence of colorectal carcinoma. Many epidemiologic data are still rather inconsistent and in some cases rather weak. Detailed case-control studies are needed to substantiate the general statement: Our life style, especially our daily food, is obviously related to the rise of colorectal carcinoma. PMID- 2984647 TI - Management of multiple sclerosis. How to improve the quality of life. AB - Multiple sclerosis manifests itself in a number of ways, affecting many body functions and systems. It is not possible to change the pathologic course of multiple sclerosis at this time, but this does not mean that physicians cannot improve the quality of life for patients with this disease. By participating in regular exercise, patients can maintain their strength and mobility. Spasticity may be controlled with medications, and urologic problems, which not only are often overlooked but also have a major bearing on patients' social and vocational life, should be controlled to the greatest extent possible. PMID- 2984648 TI - Testicular germ cell tumours--a model for a new approach to treatment of adult solid tumours. PMID- 2984649 TI - Six year remission of ACTH-dependent Cushing's syndrome using bromocriptine. AB - A patient with ACTH-dependent Cushing's syndrome remained in clinical and biochemical remission six years after pituitary irradiation and while on bromocriptine therapy. When bromocriptine was discontinued urinary free cortisol values became elevated, and were not suppressed by dexamethasone. After reintroduction of the drug, remission was again obtained. It is concluded that bromocriptine is responsible for continuing longterm remission in this case. The possible use of bromocriptine as an adjunctive therapy in ACTH-dependent Cushing's syndrome is discussed. PMID- 2984651 TI - The management of congenital hand deformities. PMID- 2984650 TI - Glutamate receptors have key role in new memory theory. PMID- 2984652 TI - [Discrepancy between the cytologic and histologic classification of bronchial cancer]. PMID- 2984653 TI - [Determination of tartrate-resistant acid phosphatase in malignant fibrous histiocytomas. Studies of routine formalin-fixed and paraffin-embedded material]. PMID- 2984654 TI - [Beethoven's disease--Paget's disease? New sources, new interpretations]. PMID- 2984655 TI - [Hematogenous metastasis of bronchial carcinoma depending on the tumor size and the presence of lymph node metastases. An autopsy study]. PMID- 2984656 TI - [Polyneuropathy and monoclonal gammopathy in Parker-Jackson tumor]. PMID- 2984657 TI - [Multiple or multicentric cerebral glioma. Diagnostic and prognostic problems]. PMID- 2984658 TI - [Determination of kinase activity by using an enzyme electrode]. AB - A method for determining the enzymatic activity of hexokinases, acetate kinase and pyruvate kinase using an enzyme electrode was developed. The assay time is 2 3 min. The lower limit of the activity determining is 0,054 U/ml. The proposed method was applied to investigation of pyruvate kinase and acetate kinase reactivation under the action of mercaptoethanol. PMID- 2984659 TI - [Hyperplasia of the thymus gland in children: physiology or pathology?]. AB - A critical analysis of the literature data and of the results of clinicoroentgenological, hormonal and immunological studies in children with thymomegaly makes it possible to regard hyperplasia of the thymus gland at early childhood as a sign of functional incompetence of the thymus and of the secondary immunodeficient state. PMID- 2984660 TI - [Comparative effects of prostaglandins and ACTH on the production of corticosterone and cyclic AMP in isolated adrenal gland cells of rats]. AB - The authors compared the effect of prostaglandin PGE2 and ACTH on corticosterone and cAMP production in isolated adrenal cells of rats. PGE2 stimulated steroidogenesis in the concentrations of 0.01-1.10 micrograms/ml and the maximum stimulation (2.5-fold) was observed in the PGE2 concentration of 0.1 micrograms/ml. ACTH stimulated steroidogenesis 12-fold as compared to PGE2. In combined addition of different PGE2 doses and the physiological ACTH dose (5 pg/ml) their steroid effects were summated. The results obtained are indicative of the fact that PGE2 is no mediator of the ACTH effect but that it is capable of potentiating this effect. Like ACTH, PGE2 raised the cAMP level in the adrenal gland. In concentrations causing a similar to ACTH increase in the cAMP level, PGE2 stimulated steroidogenesis to a lesser degree than ACTH, and in this connection a possibility of the existence of two cAMP foci in the adrenal gland is discussed. It is assumed that one of the foci is related and the other is not related with steroidogenesis, and PGE2 stimulates mainly the second focus. PMID- 2984661 TI - Genomic structure of the cottontail rabbit (Shope) papillomavirus. AB - The nucleotide sequence of the cottontail rabbit papillomavirus (CRPV or Shope papillomavirus) genome has been determined. The overall organization of the genome is similar to that of the other three papillomaviruses already sequenced. The amino acid sequences of the putative viral proteins were compared to the available protein data banks. Of particular interest is the homology found between the COOH terminus of the E2 putative protein of the different papillomaviruses and the viral or cellular mos oncogene product. This analysis also reveals a specific feature of the rabbit virus that may be related to its natural oncogenic potential. One of its open reading frames (E6) shows significant homologies with the beta subunit of a family of ATP synthases from mitochondria, chloroplasts, and bacteria, including the conservation of amino acids residues involved in nucleotide binding. The viral noncoding region includes a highly A+T-rich segment and shows a complex array of repetitions and inverted repeats that may act as control elements for gene expression and genome replication. PMID- 2984662 TI - Correlation of ouabain-sensitive ion movements with cell-cycle activation. AB - The role of tumor-promoter-induced Na+, K+-ATPase activity in cell proliferation and the extent of coupling of Na+ and K+ movements to cell-cycle control under differing physiological states was examined. Earlier studies indicated that staging of cells in G1 by serum deprivation in the presence of phorbol esters such as phorbol 12-myristate 13-acetate (PMA) induced a state(s) in which postconfluent C3H 10T1/2 fibroblasts were refractory to ouabain inhibition of DNA synthesis, and the present study further examines this property. Previous findings suggested that the promoter can act in either of two ways: (i) it can act by inducing an alternative pathway to S-phase independent of Na+,K+-ATPase activity, or (ii) the promoter can advance the cells in G1 to a point beyond which Na+, K+-ATPase activity is no longer required for the induction of DNA synthesis. When ouabain (0.3 mM) was added simultaneously with tumor promoters, such as dihydroteleocidin B, to cells arrested in the G1 phase, [3H]thymidine incorporation was inhibited greater than 90%. These data suggest that an alternative pathway is not likely the explanation but that tumor promoters advance cells through a dynamic state in G1, during which progression toward S phase entry is independent of a Na+,K+-ATPase dependent regulatory step. Ouabain sensitivity kinetics measured by two independent methods indicated that the development of ouabain insensitivity is found in G1 approximately equal to 2 hr prior to S phase. This study indicates that the measured ion movements are markedly dependent on cell-cycle state and describes the criteria required to obtain reproducible responses. PMID- 2984664 TI - Tandemly repeated nonribosomal DNA sequences in the chloroplast genome of an Acetabularia mediterranea strain. AB - A purified chloroplast fraction was prepared from caps of the giant unicellular green alga Acetabularia mediterranea (strain 17). High molecular weight DNA obtained from these chloroplasts contains at least five copies of a 10-kilobase pair (kbp) sequence tandemly arranged. This unique sequence is present in DNA from chloroplasts of all stages of the life cycle examined. A chloroplast rDNA clone from mustard hybridized with some restriction fragments from Acetabularia chloroplast DNA but not with the repeated sequence. An 8-kbp EcoRI-Pst I fragment of the repeated sequence was cloned into pBR322 and used as a hybridization probe. No homology was found between the cloned 8-kbp sequence and chloroplast DNA from related species Acetabularia crenulata or chloroplast DNA from spinach. PMID- 2984663 TI - Fine structural mapping of a critical NH2-terminal region of p60src. AB - We have recently demonstrated that an NH2-terminal sequence required for myristylation and membrane association of the Rous sarcoma virus transforming protein, p60src, is contained within amino acids 2-14 [Cross, F.R., Garber, E. A., Pellman, D. & Hanafusa, H. (1984) Mol. Cell. Biol. 4, 1834-1842]. This sequence is also required for cell transformation. We have now constructed five mutants of Rous sarcoma virus that contain alterations in the src sequence coding for these 14 amino acids. Mutants encoding src proteins with a peptide insertion between amino acids 1 and 2, or peptide substitutions for amino acids 2-4, 3-4, or 7-15, were transformation-defective. The src proteins of these mutants differed from the wild-type protein in that they were not myristylated and did not fractionate with the plasma membrane of infected cells. The fifth mutant encoded a src protein with a short peptide substituted for amino acids 11-15. This protein was myristylated and plasma membrane associated, and the virus transformed cells. We therefore conclude that a sequence required for myristylation and membrane association of p60src is located within the first 7-10 amino acids of the src protein, and that p60src myristylation and membrane association are required for cell transformation. Consistent with this idea, we have isolated four transforming revertants from one of the transformation defective mutants. The src proteins of all four revertants were found to be myristylated and membrane associated. PMID- 2984665 TI - Protonmotive force-driven active transport of D-glucose and L-proline in the protozoan parasite Leishmania donovani. AB - Midlogarithmic phase Leishmania donovani promastigotes accumulate 2-deoxy-D glucose (2-dGlc) and L-proline, maintaining concentration gradient factors across the surface membrane of 78.7 and 60, respectively. Cyanide (1 mM) and iodoacetate (0.5 mM) inhibited the transport of both substrates. L-proline uptake was also inhibited by 2-dGlc (10 mM). Transport of neither substrate was affected by Na+, phlorizin, or ouabain, indicating the sodium-independent transport of both systems. However, N',N'-dicyclohexylcarbodiimide (DCCD; 20 microM) significantly inhibited the transport of both 2-dGlc and L-proline (70% and 90%, respectively). The ionophores valinomycin (1 microM) and nigericin (5 microM) each partially inhibited the uptake of both substrates. In parallel experiments, nigericin and valinomycin were added concomitantly to promastigotes, each at a concentration that individually inhibited the transport of 2-dGlc and L-proline by less than 30%. Under such conditions, the transport of 2-dGlc and L-proline was inhibited by 69% and 78%, respectively. However, these ionophores had no significant effect on the promastigotes cellular ATP level. Carbonylcyanide p (trifluoromethoxy)phenylhydrazone (FCCP; 1 microM) inhibited 2-dGlc (79%) and L proline (85%) transport, whereas ATP levels of such cells were diminished by only 20%. Symport of D-glucose/H+ and L-proline/H+ was measured directly in cells pretreated with KCN and DCCD. Upon addition of D-glucose to such cells, a rapid movement of protons into the organisms occurred and was reversed upon addition of FCCP. Conversely, no proton movement was observed when L-glucose was added to such cells. L-proline, as D-glucose, caused a rapid influx of protons into the promastigotes, indicating that both substrates were cotransported with protons. We conclude that transport of D-glucose and L-proline in L. donovani promastigotes is protonmotive force-driven and is coupled to both delta pH and delta psi. PMID- 2984667 TI - Molecular genetics of a transposon-induced dominant mutation in the Drosophila locus Glued. AB - The organization of the Drosophila locus Glued containing the dominant allele Gl was shown to differ from that of the normal locus by an insertion of a 9-kilobase pair DNA segment near the 3' end of a transcribed region. The insertion causes the formation of a truncated polyadenylylated transcript of 5.1 kilobases instead of the normal 6.0 kilobases. The inserted DNA segment has the properties of a transposon and was identified by its corresponding restriction map as B104, which is a retrovirus-like transposon with direct terminal repeats. B104 appears to be oriented in Gl with the same polarity of transcription as Gl. The truncated Gl transcript terminates prematurely inside the 5' terminal repeat of B104, in the region of a putative polyadenylylation signal. We discuss the general implications of this finding for transposon- and retrovirus-induced mutagenesis and for the origin of dominant mutations. PMID- 2984666 TI - Platelet-derived growth factor agonist activity of a secreted form of the v-sis oncogene product. AB - We have compared the functional properties of a growth factor partially purified from medium conditioned by simian sarcoma virus-transformed cells with those of platelet-derived growth factor (PDGF). The factor mimicked the effects induced by PDGF: it bound to and activated human fibroblast PDGF receptors and stimulated DNA synthesis. These activities were specifically inhibited by PDGF antibodies and thus elicited by a factor(s) immunologically related to PDGF. The factor behaved as a secretory protein, since about 95% of the receptor-binding activity was found in the medium after a 48-hr serum-free incubation. Structural characterization of the PDGF-like activity revealed a Mr 24,000 intracellular protein and two polypeptides of Mr 13,000 and 11,500 released into the medium. The Mr 13,000 component bound to human fibroblasts; this binding was competitively inhibited by PDGF. The data support the possibility that oncogene products may elicit transforming activity by interacting with the normal cellular mitogenic pathway. PMID- 2984668 TI - Transposition of Tc1 in the nematode Caenorhabditis elegans. AB - We have identified a strain of Caenorhabditis elegans in which the transposable element Tc1 is genetically active. Most spontaneous mutations affecting the unc 54 myosin heavy chain gene of C. elegans variety Bergerac are due to insertions of Tc1 within unc-54. The Bergerac genome contains an unusually high number of Tc1 elements, but this is not responsible for transpositional activity. Another variety of C. elegans, strain DH424, contains an equally high number of Tc1 elements, but transpositions are not detected. Tc1 insertion mutations are genetically unstable. They revert to unc-54+ in both germ-line and somatic cells. Germ-line revertants are wild type and contain precise or nearly precise excisions of Tc1. Somatic revertants are genetic mosaics; they contain small patches of revertant muscle tissue in otherwise mutant animals. The pattern of mosaicism often allows us to know when and where during muscle development the excisions occur. Somatic reversion can be over 1000-fold more frequent than germ line reversion. PMID- 2984669 TI - An arylaminopyridazine derivative of gamma-aminobutyric acid (GABA) is a selective and competitive antagonist at the GABAA receptor site. AB - In view of finding a new gamma-aminobutyric acid (GABA) receptor ligand we synthesized an arylaminopyridazine derivative of GABA, SR 95103 [2-(carboxy-3' propyl)-3-amino-4-methyl-6-phenylpyridazinium chloride]. SR 95103 displaced [3H]GABA from rat brain membranes with an apparent Ki of 2.2 microM and a Hill number near 1.0. SR 95103 (1-100 microM) antagonized the GABA-mediated enhancement of [3H]diazepam binding in a concentration-dependent manner without affecting [3H]diazepam binding per se. SR 95103 competitively antagonized GABA induced membrane depolarization in rat spinal ganglia. In all these experiments, the potency of SR 95103 was close to that of bicuculline. SR 95103 (100 microM) did not interact with a variety of central receptors--in particular the GABAB, the strychnine, and the glutamate receptors--did not inhibit Na+-dependent synaptosomal GABA uptake, and did not affect GABA-transaminase and glutamic acid decarboxylase activities. Intraperitoneally administered SR 95103 elicited clonicotonic seizures in mice (ED50 = 180 mg/kg). On the basis of these results it is postulated that St 95103 is a competitive antagonist of GABA at the GABAA receptor site. In addition to being an interesting lead structure for the search of GABA ligands, SR 95103 could also be a useful tool to investigate GABA receptor subtypes because it is freely soluble in water and chemically stable. PMID- 2984670 TI - Mu and kappa opioids inhibit transmitter release by different mechanisms. AB - The actions of various opioids were examined on calcium action potentials in the cell somata of guinea pig myenteric neurones and on the release of acetylcholine at synapses onto these cells. The opioids morphine, normorphine, and [D-Ala2, MePhe4, Met5(O)]enkephalin-ol caused membrane hyperpolarizations resulting from an increase in potassium conductance; opioids that are more selective agonists for the kappa receptor subtype (dynorphin, tifluadom, U50488H) did not. Conversely, calcium action potentials were depressed or abolished by the kappa opioids but were not affected by morphine and [D-Ala2, MePhe4, Met(O)5]enkephalin ol. Both groups of opioids caused presynaptic inhibition of acetylcholine release in the myenteric plexus, depressing the amplitude of the fast excitatory postsynaptic potential. The presynaptic inhibition caused by [D-Ala2, MePhe4, Met(O)5]enkephalin-ol, morphine, and normorphine, but not that caused by the kappa opioids, was prevented by pretreatment with the selective mu site-directed irreversible antagonist beta-funaltrexamine. Furthermore, the presynaptic inhibitory action of morphine and [D-Ala2, MePhe4, Met(O)5]enkephalin-ol, but not that of the kappa-receptor agonists, was reversibly blocked by barium. The results suggest that presynaptic inhibition caused by mu receptor activation probably results from an increase in potassium conductance, whereas kappa receptor agonists may depress the release of acetylcholine by directly reducing calcium entry into the nerve terminals. PMID- 2984671 TI - Deletion of 43 amino acids in the NH2-terminal half of the large tumor antigen of simian virus 40 results in a non-karyophilic protein capable of transforming established cells. AB - We have characterized a simian virus 40 (SV40) mutant, derived from the viral DNA insertion present in simian cell transformants, which carries a deletion affecting the NH2-terminal region of the SV40 large tumor antigen. This mutant protein is 6% smaller than normal, has lost the typical nuclear localization of the SV40 large tumor antigen, and accumulates in the cytoplasm. The deletion begins at nucleotide position 4490 of the SV40 DNA and ends in-frame at nucleotide position 4362. The missing 43 amino acids begin with proline-110 and end with serine-152 of the predicted sequence; they include a cluster of basic residues, presumably important for the viral origin-DNA binding, and most of the phosphorylation sites present in the NH2-terminal half of the molecule. The protein can still be phosphorylated considerably in vivo. This mutant viral genome is replication-defective but has conserved the competence to transform established cells, such as NIH/3T3 cells. Transfection of cloned mutant DNA into such cells resulted in the production of full transformants. Full transformants were not produced in similar transfections carried out in primary rat embryo fibroblasts, although some primary transfectants expressing the non-karyophilic large tumor antigen might be considered minimally transformed. PMID- 2984672 TI - Autoregulation of collagenase production by a protein synthesized and secreted by synovial fibroblasts: cellular mechanism for control of collagen degradation. AB - Conditioned medium taken from cultures of resting rabbit synovial fibroblasts contained a protein that prevented the synthesis of the neutral proteinase collagenase. Conditioned medium was concentrated 10-fold and placed on cultures of rabbit synovial fibroblasts along with an inducer of collagenase (phorbol myristate acetate or latex particles) and [3H]leucine. Collagenase production was measured by immunoprecipitation of culture medium with monospecific antibody. Gel filtration showed that the inhibitory factor had MrS of 12,500, 25,000-50,000, and 150,000, suggesting that the protein may exist as aggregates. Activity was destroyed by boiling, by trypsin, and by dithiothreitol. Production of the inhibitory protein was prevented by cycloheximide. Isoelectric focusing purified the protein 100- to 150-fold and revealed pIs in the range of 3.2-3.7. Glycosylation was demonstrated by binding to Con A-Sepharose. Our data indicate that rabbit synovial fibroblasts autoregulate collagenase production and suggest that the low levels of collagenase seen in resting cultures result from an active suppression of collagenase synthesis. PMID- 2984673 TI - Epidermal growth factor precursor is related to the translation product of the Moloney sarcoma virus oncogene mos. AB - Murine epidermal growth factor (EGF) is synthesized as part of a large precursor (pro-EGF), which is thought to span the cell membrane. Comparison of the published pro-EGF sequence with the sequences of the translation products of viral oncogenes reveals that pro-EGF is related to the translation product of mos, the oncogene of Moloney murine sarcoma virus. Similarity is greatest between the COOH-terminal region of v-mos (residues 317-360) and part of the cytoplasmic domain of pro-EGF (residues 1127-1174). Statistical comparison of these sequences indicates that the probability of the similarity arising by chance is less than 2 X 10(-8). This similarity extends to the corresponding regions of the translation products of the cellular homologues (c-mos) of the v-mos gene present in normal murine and human DNA. Similarities are also observed between two other regions of the murine c-mos sequence (residues 48-134 and 196-275) and parts of the extracellular domain of pro-EGF (residues 565-651 and 741-817, respectively). All three mos genes are members of the tyrosine kinase family of oncogenes, as is erbB, the oncogene of avian erythroblastosis virus. Since the sequences of the erbB translation product and the EGF receptor are closely related, the relationship between mos and pro-EGF suggests that pro-EGF and the EGF receptor have evolved from a common ancestor. PMID- 2984676 TI - Tumor-promoting phorbol diesters cause the phosphorylation of epidermal growth factor receptors in normal human fibroblasts at threonine-654. AB - The effect of tumor-promoting phorbol diesters to potentiate the action of epidermal growth factor (EGF) on cell proliferation is associated with phosphorylation of EGF receptors, acute depression of EGF binding, and inhibition of EGF receptor tyrosine kinase activity. In the present studies, normal human fibroblasts and A431 carcinoma cells were labeled with [32P]phosphate and treated with and without 10 nM 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA). The EGF receptors then were isolated by immunoprecipitation and digested with trypsin. Analysis of the labeled receptor phosphopeptides by reversed-phase HPLC revealed that PMA induces the phosphorylation of a unique phosphopeptide containing [32P]phosphothreonine. Comparison of several chemical and physical properties of the 32P-labeled phosphopeptide with the primary structure of the EGF receptor suggested the identify Lys-Arg-Thr(P)-Leu-Arg. This was confirmed by direct demonstration that a synthetic peptide of this structure comigrates during HPLC and electrophoresis with the 32P-labeled phosphopeptide isolated from the EGF receptors of normal human fibroblasts. The phosphorylated site on the peptide corresponds to threonine-654 of the EGF receptor, which is located on the cytoplasmic side of the plasma membrane nine residues distant from the transmembrane domain. These data indicate that phosphorylation of the EGF receptor in human fibroblasts and A431 cells at threonine-654 may regulate the EGF receptor tyrosine kinase activity and the binding of EGF. PMID- 2984674 TI - cDNA-derived amino acid sequence of the gamma subunit of GTPase from bovine rod outer segments. AB - The sequence of the 74 amino acids in the gamma subunit of GTPase from bovine rod outer segments has been deduced from the cDNA sequence. Enriched GTPase mRNA was used to prepare a cDNA library in the expression vector lambda gt11. Clones encoding the gamma subunit were identified by screening the library with anti GTPase antibodies and by cell-free translation of hybrid-selected mRNA. The longest cDNA (448 base pairs) contained the amino acid coding region for the entire gamma subunit as well as 5' and 3' noncoding regions of 67 and 159 base pairs, respectively. The 3' noncoding region contained the sequence, A-A-U-A-A-A, the presumed recognition sequence for polyadenylylation, and a 14-unit long poly(A) tract. The amino acid sequence derived for the gamma-subunit is mostly in agreement with that previously determined by McConnell and co-workers [McConnell, D. G., Kohnken, R. E. & Smith, A. J. (1984) Fed. Proc. Fed. Am. Soc. Exp. Biol. 43, 1585] and the partial sequence deduced by Van Dop and co-workers from cDNA [Van Dop, C., Medynski, D., Sullivan, K., Wu, A. M., Fung, B. K.-K. & Bourne, H. R. (1984) Biochem. Biophys. Res. Commun. 124, 250-255]. PMID- 2984675 TI - Developmental expression of nuclear genes that encode mitochondrial proteins: insect cytochromes c. AB - To investigate tissue-specific developmental regulation of mitochondrial biogenesis, we studied the expression of the Manduca sexta (tobacco hornworm moth) thoracic muscle cytochrome c gene during adult eclosion and used this information to obtain a cDNA clone for this gene, which in turn was used to isolate the corresponding Drosophila melanogaster gene. Over the 3 days prior to adult Manduca emergence, mitochondrial inner membranes become progressively more electron dense and lamellar, and, while there is no accumulation of apocytochrome c, the amount of the holoprotein increases 40-fold per insect thorax. As determined by in vitro translation and blot hybridization analysis, the major thoracic muscle cytochrome c gene is primarily regulated at the transcriptional level, with cytochrome c mRNA increasing from less than 0.01% to 0.04% of total poly(A)+ RNA and declining to an undetectable level by day 2 after eclosion. Furthermore, the ratio of cytochrome c to the other cytochromes remains the same at all times, indicating that these components of the respiratory chain follow coordinated developmental programs. By using polysome immunoadsorption, a poly(A)+ RNA population of greater than or equal to 95% cytochrome c mRNA was isolated from thoracic muscle tissue and was used to construct a cDNA library, which was screened by hybrid selection/translation. We report the sequence of one of those clones, pMSc750, and its use to isolate the major thoracic muscle cytochrome c gene of Drosophila. PMID- 2984677 TI - 1H NMR studies of electron exchange rate of Pseudomonas aeruginosa azurin. AB - T1 values of the His-35 C-2 proton resonance of reduced Pseudomonas aeruginosa azurin were determined at 25 degrees C and pH values 4.5, 7.3, and 9.0 in the presence of different fractional amounts of oxidized azurin. The C-2 proton of His-35 undergoes very rapid spin relaxation in oxidized azurin because of its close proximity to the paramagnetic copper. In the presence of oxidized protein, the T1 values of this proton in reduced azurin depend on the lifetime of the reduced protein. From the T1 data, the electron self-exchange rate constant for azurin was calculated to be 1.4 X 10(4) M-1 X s-1, 4.3 X 10(3) M-1 X s-1, and 6.0 X 10(3) M-1 X s-1 at pH values 4.5, 7.3, and 9, respectively. At pH 7.3, the C-2 proton of His-35 is in slow exchange between the imidazole and imidazolium forms and gives rise to two separate resonances at 9.39 and 8.00 ppm. By using these two resonances, the electron self-exchange rate constants were determined separately for the two species of azurin for which the His-35 residue is in the imidazole or the imidazolium forms; results showed that both species participate in self-exchange of electrons with equal efficiency. PMID- 2984678 TI - Rat Sertoli cells acquire a beta-adrenergic response during primary culture. AB - Two-dimensional polyacrylamide gel electrophoresis and the radioligand (-) [125I]iodopindolol (125I-Pin) have been used to study isoproterenol-dependent protein phosphorylation and beta-adrenergic receptor availability, respectively, in cultured Sertoli cells and freshly isolated seminiferous tubular segments of sexually immature and mature rats. Sertoli cells prepared from sexually immature rats show progressive 125I-Pin binding in primary cultures that correlates with isoproterenol-induced cell shape changes, redistribution of immunoreactive vimentin, and phosphorylation of this intermediate filament protein. The development of 125I-Pin binding to Sertoli cell lysates is blocked by cycloheximide. Seminiferous tubules do not show significant isoproterenol dependent vimentin phosphorylation nor 125I-Pin binding. However, vimentin phosphorylation can be induced by follicle-stimulating hormone or a cyclic nucleotide analog. This study stresses the need for correlating pharmacological induced responses observed in Sertoli cell primary cultures with those in the intact seminiferous tubule. PMID- 2984679 TI - Transposon-like sequences in extrachromosomal circular DNA from mouse thymocytes. AB - Small polydisperse circular (spc) DNA was isolated from mouse thymocytes and cloned into the HindIII site of lambda vector Charon 7. Fifty-six recombinants from this spc DNA library were analyzed. R repeats, which were originally found near immunoglobulin genes, were enriched in spc DNA clones relative to their representation in the chromosome. In one clone, the R sequence was linked to Bam and MIF sequences and the contiguous arrangement was truncated from both ends. In another clone, composite Bam/R and R repeats existed as a pair in inverted repeat orientation. Truncation occurred from the 5' side without affecting the 3' ends. In both clones, short direct repeats flanked the repeated sequences. The possible role of R sequences in transposition and circular formation is discussed. PMID- 2984680 TI - Mechanisms determining aerobic or anaerobic growth in the facultative anaerobe Salmonella typhimurium. AB - We isolated mutant strains of the facultative anaerobe Salmonella typhimurium that grow either aerobically or anaerobically. Strict anaerobic mutants contained a defective DNA topoisomerase I gene (topI), while strict aerobic mutants contained a defective DNA gyrase subunit A gene (gyrA, also nalA). Topoisomerase I activity was detected in cell-free extracts of strict aerobic mutants but not of strict anaerobic mutant strains, whereas gyrase activity was detected in extracts of strict anaerobic mutants but not of strict aerobic mutants. Furthermore, extracts of wild-type cells, cultured under vigorous aerobic condition, contain topoisomerase I activity but no significant gyrase activity. In contrast, the extracts of anaerobically cultured wild-type cells contain gyrase activity but no significant topoisomerase I activity. Sucrose gradient centrifugation with ethidium bromide showed that chromosomal DNA in strict aerobic mutants and aerobically grown wild-type cells was relaxed, while the chromosomal DNA of strict anaerobic mutants and anaerobically grown wild-type cells was more supercoiled. Aerobic cultures of wild type and strict aerobic mutants produced both superoxide dismutase and catalase, whereas anaerobic cultures of wild type and strict anaerobic mutants did not. These results lead us to conclude that activity of topoisomerase I, associated with relaxation of chromosomal DNA, is necessary for expression of genes required for aerobic growth, whereas activity of gyrase, associated with supercoiling of chromosomal DNA, is necessary for expression of genes required for anaerobic growth. PMID- 2984681 TI - DNA sequences at the ends of the genome of bacteriophage Mu essential for transposition. AB - We have determined the minimal DNA sequences at the ends of the genome of bacteriophage Mu that are required for its transposition. A mini-Mu was constructed on a multicopy plasmid that enabled the manipulation of the DNA sequences at its ends without affecting the genes essential for transposition. The genes A and B, which were cloned outside the ends of the mini-Mu on the same plasmid, were both needed for optimal transposition. In our experimental system the predominant end products of the transposition are cointegrates both in the presence and in the absence of B. Two regions ending approximately 25 and 160 bp from the left end and one ending approximately 50 bp from the right end appear to be essential for optimal transposition. Overlapping with these regions, a 22-base pair sequence was recognized with the consensus Y-G-T-T-C-A-Y-T-N-N-A-A-R-Y-R-C-G A-A-A-A, where Y and R represent any pyrimidine and purine, respectively. At the left end these sequences occur as direct repeats; at the right end this sequence is inverted with respect to those at the left end. PMID- 2984682 TI - Monoclonal human thyroid cell line GEJ expressing human thyrotropin receptors. AB - By using the hybridoma technology, a monoclonal human thyroid cell line was obtained by fusing fresh normal human thyroid cells with a human lymphoblastoid cell line. The resultant cell line, called GEJ, has been selected for its expression of thyrotropin (TSH) receptors and has morphological and functional characteristics of normal human thyroid cells. In the presence or absence of human TSH, the GEJ cell line accumulates iodide, produces thyroid hormones, expresses thyroid membrane antigens, and binds approximately equal to 600 molecules of TSH per cell. The binding site for TSH has all the characteristics of specific receptor (i.e., temperature and time dependencies, dissociation of bound TSH only by high amounts of TSH, and a glycoprotein nature). Scatchard analysis described a curvilinear graph with two dissociation constants (Kd = 0.12 X 10(-9) M and 1.6 X 10(-9) M) with, respectively, 1.2 X 10(3) and 7.2 X 10(3) binding sites per cell. This human thyroid cell line that expresses TSH receptors could be a useful tool for the study of human thyroid disorders. PMID- 2984683 TI - Production of antibody associated with non-A, non-B hepatitis in a chimpanzee lymphoblastoid cell line established by in vitro transformation with Epstein-Barr virus. AB - A continuous cell line of chimpanzee lymphocytes producing an antibody specifically associated with non-A, non-B hepatitis (NANB) was established. Peripheral blood lymphocytes of a chimpanzee convalescent from experimental infection with NANB hepatitis were transformed in vitro by Epstein-Barr virus infection into lymphoblastoid cell lines. Supernatants of the cell cultures were screened by immunofluorescence for antibody activity against the liver tissue of a chimpanzee with NANB hepatitis. Nineteen of the 1402 cultures were found to be positive for the activity. Ten of these 19 gave cytoplasmic reactions and the remaining 9 gave nuclear reactions in hepatocytes. One culture (48-1) stably producing the antibody was further characterized. The antibody produced in 48-1 was IgM and gave granular cytoplasmic reactions in hepatocytes. Cloning of 48-1 was performed by the soft agar method and cloned cell lines stably producing the antibody were obtained. The 48-1 antibody reacted with liver biopsy specimens from 12 chimpanzees obtained during the acute or chronic phase of hepatitis caused by five different NANB strains, but not with biopsy specimens from chimpanzees with hepatitis A or B or from normal chimpanzees. In addition, examinations of serial liver biopsy specimens obtained from 2 chimpanzees experimentally infected with NANB hepatitis demonstrated that the antibody reacted with the biopsies obtained during the preacute, acute, and chronic hepatitis, but not with those obtained before inoculation, early incubation period, or during convalescence. The present results indicate the specific association of the antibody with NANB hepatitis. Immunoelectron microscopy revealed that the antibody reacted with the microtubular aggregates identical to those previously described in a patient and chimpanzees with NANB hepatitis. PMID- 2984685 TI - Nutrition and carcinoma of the large intestine. PMID- 2984684 TI - Sequence analysis of hepatitis A virus cDNA coding for capsid proteins and RNA polymerase. AB - We report here the nucleotide sequence corresponding to two large regions of the hepatitis A virus (HAV) genome. These comprise a sequence of 3274 bases corresponding to the 5' end of the genome, which includes the putative capsid protein region of this picornavirus, and 1590 bases corresponding to the 3' end of the genome, terminating in a 15-base poly(A) tract. These sequences revealed that HAV had the characteristic genomic organization of picornaviruses: an open reading frame beginning approximately 750 bases from the 5' end of the RNA and a termination codon 60 bases from the 3' poly(A) tract. The predicted amino acid sequences of both regions have been compared to analogous regions previously determined for other picornaviruses. There was sufficient homology to conclude that the 5' region of HAV codes for capsid proteins and that the 3' region codes for an RNA polymerase. However, these regions of HAV were not found to be closely related to analogous regions of poliovirus, encephalomyocarditis virus, and foot and mouth disease virus. PMID- 2984686 TI - Nutrition and cholesterol gallstones. PMID- 2984687 TI - The irritable bowel controversy. PMID- 2984688 TI - The potential of ruminants for the utilization of fibrous low-quality diets. PMID- 2984689 TI - Splenic leukocytes from chickens injected with Salmonella pullorum antigen stimulate production of corticosteroids by isolated adrenal cells. AB - Corticosteroid levels in the blood serum of White Rock pullets were significantly increased within 1 hr after an iv injection of heat-killed Salmonella pullorum antigen (SP-Ag), and reached levels 7 to 10 times that of PBS-injected controls within 2 hr after SP-Ag injection. Incubations with isolated adrenal cells indicated that serum from SP-Ag-injected birds had the ability to stimulate the synthesis or release of corticosteroids twice that of serum PBS-injected birds. Stripping the serum from SP-Ag-injected birds with activated charcoal and precipitated silica (Quso G-32) removed the corticosteroids and the adrenal stimulating ability. A 2-hr incubation of isolated adrenal cells with leukocytes from spleens removed from chickens 1 hr after injection with SP-Ag, using stripped serum as the medium, stimulated a two- to fivefold increase in corticosteroid as compared to splenic leukocytes from PBS-injected chickens incubated in the same medium. The results indicate that an "ACTH-like" substance was produced by the S. pullorum antigen-stimulated splenic leukocytes. PMID- 2984690 TI - Safety of human blood products: inactivation of retroviruses by heat treatment at 60 degrees C. AB - Acquired immune deficiency syndrome (AIDS) can be transferred to patients by blood transfusions or human blood preparations, such as cryoprecipitates or factor VIII concentrates. Retroviruses have been discussed as infectious AIDS agents and more recently human T-lymphotropic retroviruses designated as HTLV type III and LAV (lymphadenopathy-associated virus) have been isolated from AIDS patients. Whether heat treatment at 60 degrees C (pasteurization) of liquid human plasma protein preparations inactivates retroviruses was therefore investigated. Pasteurization had already been included in the routine manufacturing process of human plasma protein preparations in order to guarantee safety with regard to hepatitis B. Since high titer preparations of human retroviruses were not available, heat inactivation was studied using Rous sarcoma virus added to the various plasma protein preparations tested. This retrovirus which was obtained in preparations of 6.0 log10 FFU/ml was shown to be at least as heat stable as two mammalian retroviruses studied, i.e., feline and simian sarcoma virus. In all of eight different plasma protein preparations tested, Rous sarcoma virus was completely inactivated after a heat treatment lasting no longer than 4 hr. It is thus concluded that pasteurization of liquid plasma protein preparations at 60 degrees C over a period of 10 hr must confer safety to these products with respect to AIDS, provided that the AIDS agents are retroviruses of comparable heat stability as Rous sarcoma virus and the mammalian retroviruses tested. PMID- 2984691 TI - Adrenocortical function in deoxycorticosterone acetate (DOCA)-hypertensive Yucatan miniature swine. AB - Adrenocortical function was assessed in six normal and six chronic (greater than 12 weeks), DOCA-hypertensive Yucatan miniature swine; mean arterial pressures were 115.3 +/- 11.7 and 163.6 +/- 27.2 mm Hg, respectively (mean +/- SEM). Adrenocortical function was evaluated in vivo by measuring changes in plasma cortisol and aldosterone in response to exogenous ACTH (0.25 mg, iv), and in vitro by measuring the responses of collagenase-isolated adrenocortical cells to ACTH and angiotensin II. Corticoids were measured by specific radioimmunoassay. Basal plasma cortisol values of conscious DOCA-hypertensive swine were approximately 53% of the values of normotensive swine (P less than 0.05). However, ACTH induced a 419% increase in plasma cortisol values in DOCA hypertensive swine compared to a 261% increase in the normotensive swine (P less than 0.05). These differences between the two groups were not altered by anesthesia. There were no significant differences in ACTH-induced changes in plasma aldosterone between the normotensive and DOCA-hypertensive swine. Experiments in vitro showed that the corticoid secretory responses of adrenocortical cells from DOCA-hypertensive animals were 6 times more sensitive to ACTH and 3.2 times more sensitive to angiotensin II than those of cells from normotensive swine. Thus, despite the possibility of adrenocortical insufficiency due to suppressed plasma renin activity and the negative feedback of DOCA on the hypothalamic-hypophyseal-adrenal axis, adrenocortical function of DOCA hypertensive swine was hyperresponsive to trophic hormones. Results from this study suggest that the DOCA-hypertensive swine may be a valuable model in elucidating the relationship between hypertension and adrenocortical function and in investigating nonclassical control of the adrenal cortex, that is, control exerted during the hypertensive state that exists apart from or in addition to that exerted by ACTH and angiotensin II. PMID- 2984692 TI - Effect of (E)-5-(2-bromovinyl)-2'-deoxyuridine on the growth and viral capsid antigen expression of Epstein-Barr virus-associated tumor (B-95-8) cells transplanted to nude mice. AB - When persistently Epstein-Barr virus (EBV)-infected lymphoblastoid (B-95-8) cells were transplanted subcutaneously or intracerebrally to nude mice of either BALB/c or NIH background, tumors developed, and the tumor cells spontaneously expressed viral capsid antigen (VCA). This model was used to evaluate the in vivo anti-EBV activity of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU), a highly potent and selective antiherpes agent, which was recently shown to inhibit several parameters of EBV infection in vitro. When administered intraperitoneally at 200 mg/kg/day for 4 weeks, or 500 mg/kg/day for 2 weeks, starting immediately after B 95-8 cell inoculation, BVDU effectively reduced tumor growth and VCA expression of either subcutaneously or intracerebrally inoculated B-95-8 cells. PMID- 2984693 TI - Prolonged relaxation of detergent-skinned smooth muscle involves decreased endogenous phosphatase activity. AB - Since contraction of smooth muscle involves Ca2+-dependent phosphorylation of the 20 Kd myosin light chains, changes in endogenous phosphatase activity may participate in regulating smooth muscle contractility. We found that detergent skinned fibers from 7 of 10 chicken gizzards studied were characterized by relatively high endogenous light chain phosphatase activity (23 mU/mg protein) and rapid relaxation (t1/2 = 1-3 min) in the absence of Ca2+ (less than 10(-8) M). In contrast, skinned fibers from 3 of the gizzards exhibited very low phosphatase activity (3 mU/mg protein) and markedly prolonged relaxation (t1/2 = 50-200 min). However, such slow relaxing fibers were converted to a form resembling rapidly relaxing fibers (t1/2 = 4-10 min) when an aortic polycation modulable phosphatase was included in the incubation medium. Moreover this phosphatase-enhanced relaxation was associated with dephosphorylation of the light chains. Maximal isometric force (1 mN) and light chain phosphorylation (0.8 mol PO4/mol light chain) were similar in slowly and rapidly relaxing fibers. Thus, the two populations of skinned fibers, though dramatically different with respect to phosphatase activity and relaxation time, appeared to be very similar in terms of Ca2+-dependent contraction. These findings strongly suggest that prolonged relaxation of smooth muscle of the kind noted in this study, and perhaps in hypertensive or aging vascular smooth muscle, may reflect decreased endogenous phosphatase activity. PMID- 2984695 TI - Early and late developmental abnormalities associated with congenital cytomegalovirus and toxoplasma infections. PMID- 2984694 TI - Retrovirus-like particles in salivary glands, prostate and testes of AIDS patients. AB - AIDS associated retrovirus-like particles were identified in the salivary gland, prostate and/or testicle of two AIDS patients. These findings further suggest that saliva and semen may transmit the infection to susceptible individuals. PMID- 2984696 TI - Preventing neonatal herpes simplex infections: an obstetrician's perspective. PMID- 2984698 TI - Neural differentiation in embryonal carcinoma cells. PMID- 2984697 TI - A possible purification of mullerian inhibiting substance and a model for its mechanism of action. PMID- 2984699 TI - Pepsinogen synthesis and secretion by isolated gastric glands. AB - De novo synthesis of pepsinogens was demonstrated in gastric glands isolated from rabbit and human gastric mucosa. The isolated glands were incubated in an amino acid free, Minimum Eagles Medium supplemented with a 14C labelled amino acid mixture. The glands were centrifuged at different time intervals and aliquots of gland homogenates and medium were run on polyacrylamide slab gels. Newly synthesized pepsinogens were demonstrated by autoradiography. Incorporation of 14C was detected after 30 minutes of culture and increased almost linearily with time over 4 h. By comparing the electrophoretic patterns after autoradiography, protein and pepsinogen activity staining, it was concluded that the glands synthesize mainly pepsinogens. Cimetidine, at a concentration of 10(-4)M strongly inhibited pepsinogen synthesis. Spontaneous secretion of pepsinogens into the medium was very low and relatively constant. Cyclic dibutyryl AMP markedly stimulated the secretion of pepsinogens into the medium. The results show, that isolated gastric glands are capable of synthesis and secretion of pepsinogens and that each function can be stimulated and inhibited selectively. PMID- 2984700 TI - Chronic naltrexone treatment increases the heroin-produced facilitation of self stimulation. AB - The facilitatory effects of heroin HCl (0.25 mg/kg, SC) on self-stimulation (SS) of the lateral hypothalamus before and after chronic treatment of naltrexone (10 mg/kg, SC, for 20 days) or vehicle were compared. The group that received chronic naltrexone had a larger heroin-induced facilitation of SS than the group that received vehicle. These data suggest that the sensitivity to the facilitatory effect of heroin on SS may be related to the amount of opiate receptor binding which is increased following chronic antagonist treatment. However, neither acute nor chronic treatment with naltrexone produced any significant changes in SS thresholds, suggesting that the directly stimulated substrate for the rewarding effect of brain stimulation is unlikely to be endorphinergic but is apparently modulated by the endogenous opioid system. PMID- 2984701 TI - Effects of vasopressin on noradrenaline-induced cyclic AMP accumulation in rat brain slices. AB - Addition of arginine vasopressin (AVP) or 1-desamino-8-D-arginine vasopressin (DDAVP) to rat cortical slices resulted in significant inhibition of the rise in cyclic AMP produced by incubation with 50 microM noradrenaline. A single injection of DDAVP (20 micrograms/rat) produced a reduced response to noradrenaline in derived cortex and caudate slices. In animals pretreated at day 5 of life with IP desipramine and intracisternal 6-hydroxydopamine (6-OHDA), both acute and chronic treatments with DDAVP resulted in a reduction in response in derived cortical, caudate and hippocampal slices. The 6-OHDA pretreated animals also showed reduced open-field behavioural activity after both acute and chronic DDAVP, while animals which were not pretreated responded to acute treatment only. The relationship between the effects of vasopressin on noradrenaline-induced cyclic AMP accumulation and its action on learning and memory is discussed. PMID- 2984702 TI - Behavioral response to apomorphine and its interaction with opiates in domestic pigeons. AB - Domestic pigeons received peripheral injections of saline or the dopamine agonist apomorphine (AM) at doses of 0.025, 0.05, 0.1, 0.25, 0.5 or 1 mg and their behavior was studied for 30 min after these treatments. Given at a dose of 0.025 mg, AM decreased pecking, whereas doses ranging from 0.1 to 1 mg strongly stimulated this behavior. The frequency of headshaking was enhanced by the administration of each dose of AM; at the 3 higher doses, the drug also attenuated the frequency of preening. In another experiment, AM was administered 40 min after the injection of either naloxone (0.5, 1 or 4 mg), the opiate agonist levorphanol (0.25, 0.5 or 1 mg) or its dextroisomer, dextrorphan (0.25, 0.5 or 1 mg), while the birds were observed as before. No interaction between AM and either naloxone or dextrorphan was detected. By contrast, injection of each dose of levorphanol attenuated preening, and completely antagonized the stimulating effect of AM treatment on headshaking. At a dose of 1 mg, levorphanol also slightly decreased the frequency and increased the latency of occurrence of pecking. It is concluded that in pigeons, opiates modulate the behavioral response to apomorphine in a complex fashion. PMID- 2984703 TI - Differences of alteration in opioid systems induced by conditioned suppression and electric footshock in mice. AB - The motility, pain-threshold and opioid receptor activities of the synaptic membrane in mice showing conditioned suppression of motility were compared with those in mice given only electric footshock. Electric footshock caused analgesia and a decrease in motility, both of which were partially reversed by administration of high doses of naloxone. In contrast, mice exhibited a marked suppression of motility (conditioned suppression) but not analgesia when placed in the same environment 24 hr after the electric footshock in which the animals received the electric footshock. In the electric footshock group, the [3H] naloxone binding capacity at low affinity site was increased. These results suggest that the increase in [3H]-naloxone binding capacity may play an important role in the behavioral changes of electric footshock group, but not conditioned suppression group. PMID- 2984704 TI - Receptor stereospecificity in opiate-ethanol interaction using the preexposure conditioned taste aversion (CTA) paradigm. AB - In the first experiment, rats were conditioned with different doses of levorphanol or dextrorphan to a novel tasting saccharin solution. In the second experiment, rats were either preexposed to levorphanol or dextrorphan and conditioned with either morphine or ethanol to the saccharin solution. The results showed that levorphanol, but not dextrorphan, at 1, 5 and 10 mg/kg doses effectively induced a CTA. Preexposure to the 5 mg/kg dose of levorphanol blocked both morphine- and ethanol-induced CTAs. Dextrorphan at the same dose did not affect the CTAs. These findings are discussed in terms of the involvement of the opiate receptors in opiate-ethanol interaction. PMID- 2984705 TI - Effects of nicotine on beta-endorphin, alpha MSH, and ACTH secretion by isolated perfused mouse brains and pituitary glands, in vitro. AB - The effects of nicotine on secretion of the pituitary peptides beta-endorphin, alpha MSH, and ACTH were studied using the isolated perfused mouse brain (IPMB) and isolated superfused pituitaries of C3H mice. Nicotine (6.1 microM) stimulated secretion of beta-endorphin immunoreactivity from C3H IPMB approximately twofold. Secretion of alpha MSH immunoreactivity was stimulated approximately two- and sixfold by 6.1 microM and 12.2 microM nicotine, respectively. However, nicotine (6.1 microM) had no direct effect on the secretion of beta-endorphin, alpha MSH, or ACTH immunoreactivities from the isolated superfused pituitaries. The data suggest nicotine acts in the brain to stimulate pituitary secretion of alpha MSH and beta-endorphin. Electrocorticographic (ECoG) activity of the IPMB was monitored. Nicotine induced characteristic ECoG changes including a reduction of input voltage, a biphasic response of rapid desynchronization followed by prolonged synchronization, and seizure at high doses (12.2 microM). PMID- 2984706 TI - Changes of HDL-lipid composition as related to delta 9-THC action. AB - An attempt was made to investigate the possible action of delta 9-THC on HDL Lipid composition. Significant changes were observed in the serum total lipids, triglycerides and HDL subfractions after hashish smoking. The results are discussed in relation to the possible alterations of some enzymatic mechanisms regulating lipid metabolism in hashish users. PMID- 2984708 TI - Enhancement of diazepam activities induced by denzimol in mice. AB - Denzimol, a new anticonvulsant drug, enhances the depressant and antimetrazol activities of diazepam in mice, in a dose and time-dependent fashion. The depressant and anticonvulsant activities of phenobarbital were not affected by Denzimol. It is suggested that Denzimol induces an increase in the number of the benzodiazepine receptors, and that such increase might be responsible for the enhancement of the diazepam's activities. PMID- 2984707 TI - Captopril inhibits sodium and water transport in the toad skin, a model of the distal nephron. AB - Captopril (SQ 14225), an orally active angiotensin I-converting enzyme inhibitor (CEI), increases natriuresis and diuresis in man and experimental animals in vivo, as well as in the isolated perfused rat kidney, raising the possibility of a direct renal action of the drug. We tested this hypothesis by studying its effects in the isolated toad skin, a model of the distal nephron devoid of vascular and nervous influences. When added to the dermal bath, captopril caused a reversible, concentration-related decrease in short-circuit current (SCC), a measure of active transepithelial Na transport. Keeping the toads in 0.1 M NaCl for 4 or more days increased sensitivity to the drug, which then inhibited SCC maximally (49 +/- 12% at 3.4 X 10(-3) M, P less than 0.01, n = 10), suggesting its effect might be modulated by endogenous mineralocorticoid activity. Captopril also inhibited the increase in SCC and in osmotic water permeability caused by neurohypophyseal peptides (NHP). The increases in SCC by non-peptidic agents (nystatin, a polyene antibiotic, or norepinephrine, an adrenergic agonist) were not altered, ruling out a generalized toxic effect, or any significant inhibition of the Na pump by captopril. The apparently specific effect of the drug on the permeability responses to NHP seems to be exerted proximally to the apical border, since the response of the latter to other agents was preserved. The present data suggest SH groups may be involved, since other CEI lacking such groups (teprotide and MK-422) do not produce such effects. These observations support the notion that a direct tubular effect may be involved in the increased diuresis and natriuresis observed after administration of captopril. PMID- 2984709 TI - Anorectic effect of fenfluramine isomers and metabolites: relationship between brain levels and in vitro potencies on serotonergic mechanisms. AB - A study of the possible molecular mechanisms of action by which the isomers and metabolites of fenfluramine increase serotonin transmission, leading to anorectic activity, is presented. The actual brain levels of fenfluramine and norfenfluramine isomers after administration of equi-anorectic doses to rats are compared with their potencies in affecting serotonergic mechanisms in vitro. Isomers and metabolites of fenfluramine can have the same pharmacological action by influencing serotonin uptake, release and binding in a quantitatively different manner. PMID- 2984710 TI - Behavioral pharmacokinetics of marijuana. AB - Male volunteer subjects smoked one marijuana cigarette containing 100, 200, or 250 micrograms/kg delta-9-tetrahydrocannabinol (THC) and were tested on three perceptual-motor performance measures related to driving. Performance was measured and blood samples were collected for 24 h after smoking. The covariation between pharmacodynamics of performance and pharmacokinetics of THC in plasma was investigated for decrement in performance as the response to smoking a single marijuana cigarette. A significant linear correlation was found between tracking errors under divided attention and THC plasma levels over 5-25 ng/ml for approximately 2 h after smoking. A sigmoid relation was found between critical tracking breakpoint and log THC plasma levels over 2-25 ng/ml for approximately 7 h after smoking. PMID- 2984712 TI - Notes on the consensus development process. PMID- 2984711 TI - Effects of naloxone, beta-endorphin and ACTH on acquisition of schedule-induced polydipsia. AB - A series of three experiments examined the possible involvement of endogenous opioid peptides in the development of schedule-induced polydipsia in rats. Repeated pretraining treatment with 2 mg/kg naloxone impaired acquisition of schedule-induced polydipsia, whereas the same treatment injected after training increased drinking. This later effect was time dependent, since a 30-min delay in the injection of naloxone resulted in a disappearance of its effect. Post training injections of 10 micrograms/kg beta-endorphin or ACTH delayed the development of drinking. These findings are consistent with the hypothesis that endogenous opioid peptides modulate the development of schedule-induced polydipsia. PMID- 2984713 TI - Serotonin agonists and beta-adrenergic agonists as a treatment for depressive disorder: a direct clinical application. PMID- 2984714 TI - An NIMH view of consensus conferencing. PMID- 2984716 TI - Seventeen-year mortality experience of proton radiation in Macaca mulatta. AB - This is an interim report on the lifetime study of chronic mortality and its causes under investigation in 31 control (20 males, 11 females) and 217 survivors (124 males, 93 females) of an acute 90-day experiment in rhesus monkeys. Single acute whole-body exposures were made using 32-, 55-, 138-, 400-, and 2300-MeV protons in 1964-1965. Doses ranged from 25 to 800 rad and dose rates from 12.5 and 100 rad per minute. Tissue depths of partially penetrating 32- and 55-MeV particles were approximately 1 and approximately 2.5 cm, respectively, and depth doses at the respective distances were 115 and 122% of surface doses. Protons with energies greater than or equal to 138 MeV were totally penetrating and the depth doses were essentially homogenous. For pooled data: (1) mortality was significantly higher (P less than 0.01) in irradiated animals (48%) than in controls (19%); (2) mortality in animals exposed to partially penetrating 55-MeV protons (53%) was essentially similar to those given totally penetrating 138- (53%), 400- (49%), and 2300-MeV (44%) exposures; (3) proton energies and doses that were effective in producing life shortening were greater than or equal to 55 MeV and greater than or equal to 360-400 rad, respectively; (4) death rates for irradiated animals compared to controls began to increase after approximately 8 years, approximately 2 years, and approximately 1 year for those exposed to 360 400, 500-650, and 800 rad, respectively; (5) of the nine probable causes of death reported, the leading causes were primary infections in both irradiated (31%) and control (50%) animals, endometriosis (25% vs 0%, respectively), neoplasms (17% vs 0%), and organ degeneration (17% vs 33%); and (6) if endometriosis is included with the neoplastic group, deaths from all forms of neoplasms would be 42% in irradiated animals.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984715 TI - Biodistribution of the radioprotective drug 35S-labeled 3-amino-2-hydroxypropyl phosphorothioate (WR77913). AB - 3-Amino-2-hydroxypropyl phosphorothioate (WR77913), a less toxic phosphorothioate radioprotector than WR2721, has been labeled with 35S. The biodistribution of a radioprotective dose of 800 mg/kg was determined in C3H mice bearing RIF-1 tumors as a function of time after intraperitoneal injection and was expressed as percentage injected dose/gram (% ID/g). Levels of 35S in the blood peaked 10 min after injection, and radioactivity in most tissues was highest at 15 min. Label in most tissues declined markedly between 15 and 60 min, but in gut, salivary glands, tumor, and brain, the levels of radioactivity remained quite stable over 1 hr. At 30 min after injection the highest levels of labeled drug were found in submandibular salivary glands, gut, and kidney, with the lowest level in brain. Tumors had approximately the same amount of label as blood, muscle, skin, and esophagus. Two principal differences between the distribution of label from WR77913 and WR2721 were defined. Although blood levels of 35S-WR2721 also peaked 10 min after injection, the 10-min blood levels achieved for WR77913 were more than fourfold greater than those attained by WR2721. Maximum levels of WR2721 occurred in most tissues 30 to 60 min after administration of the drug, compared to 15 min for WR77913. The basis for these differences remains to be determined, but these results suggest that the optimum interval between administration of WR77913 and irradiation may be shorter than for WR2721. PMID- 2984717 TI - Comparison of cAMP with other radioprotectors against chronic damage to the rat parotid gland. AB - Radiation damage to the parotid gland is protectable by cAMP during the first week after irradiation (acute phase), though appreciable recovery occurred later with or without such protection. Further damage developed later (chronic phase, 60-90 days), and cAMP was still protective against this damage with a dose modification factor of 1.86 for gland weight. A summary of the protective factors, acute and chronic, for WR-2721, isoproterenol, and cAMP is included. Chronic damage is about 1.5 times as great as acute, and protection against acute and chronic damage is about equal for all three compounds. PMID- 2984718 TI - Basilar venous plexus of the posterior fossa: a potential source of error in petrosal sinus sampling. AB - Sampling of serum from the inferior petrosal sinus can provide important information about the source of elevated adrenocorticotropic hormone (ACTH) levels. This often leads to improved results of pituitary surgery for Cushing disease. The authors describe a successful catheterization technique and illustrate the venous anatomy of the inferior petrosal sinuses and basilar plexus. PMID- 2984719 TI - Hepatic tumors: differentiation by transverse relaxation time (T2) of magnetic resonance imaging. AB - Fifty-three patients who had hepatic tumors (24 hepatomas, ten metastases, and 19 cavernous hemangiomas) underwent MR imaging using a 0.35-T superconducting imager. The transverse relaxation time (T2) was calculated from a pair of spin echo images (repetition time [TR] of 1600 msec) with echo delay times (TE) of 35 and 70 msec. The computed T2 value was obtained in a fashion similar to that used to obtain CT numbers with region-of-interest cursors. The mean T2 was 59 +/- 9 msec in hepatomas, 64 +/- 15 msec in metastases, and 100 +/- 30 msec in hemangiomas. The difference between the T2 of hemangioma and that of liver malignancies was statistically significant (P less than .001); however, differentiation between hepatoma and metastases was not possible. The T2 was shorter than 80 msec in all 24 hepatomas and in nine of ten metastases, and was longer than 80 msec in 16 of 19 hemangiomas. Forty-nine of 53 cases (92%) were correctly classified when the borderline of T2 between hemangioma and hepatic malignancies was set at 80 msec. MR with T2 calculation was valuable in differentiating between hemangioma and hepatic malignancies. PMID- 2984720 TI - Focal and diffuse liver disease studied by quantitative microstructural sonography. AB - Quantitative microstructural sonography differs from conventional sonography in several ways. The frequency as well as the amplitude of echoes is recorded and assessed using spectral analysis techniques. The effects of diffraction of the ultrasound beam are calibrated, as are the transfer characteristics of the system. Studies of patients with alcoholic liver disease revealed significantly increased attenuation, heterogeneity of scatterers, and local scattering strength in those with severe disease; in contrast, patients with mild cirrhosis exhibited decreased local scattering strength compared to normal individuals. Using retrospective three-parameter discriminant analysis, 23 out of 24 patients were successfully categorized, suggesting that the parameters used were capable of measuring microstructural changes associated with these disease states. Parameters varied widely among 6 types of hepatic tumors, suggesting that this method may offer promise in distinguishing tumors from normal cases and alcoholic liver disease as well as from each other. Further applications and evaluation of this method appear justified. PMID- 2984721 TI - Hepatocellular carcinoma: US evolution in the early stage. AB - To study the sonographic evolution of hepatocellular carcinoma (HCC) in its early stage, 26 HCCs in 24 patients were observed regularly with real-time ultrasound for a period of 90 to 691 days. In the beginning, 21 tumors were hypoechoic, two isoechoic, and three diffusely hyperechoic. On follow-up, two of the 21 initially hypoechoic HCCs remained the same echodensity, 12 increased in internal echoes but were still hypoechoic, one became isoechoic, four changed to hyperechoic, and the remaining two shifted from hypoechoic to isoechoic and then to hyperechoic. The two initially isoechoic HCCs also gained echogenicity and became hyperechoic. By contrast, the three initially hyperechoic HCCs kept the same echo patterns. The "acquired" hyperechoic HCCs were inhomogeneous in echodensity and larger in size whereas the three originally hyperechoic HCCs were homogeneous and smaller. It is concluded that most small HCCs evolve progressively from hypoechoic to isoechoic and then to inhomogeneously hyperechoic patterns as they grow; a few HCCs have diffusely high echogenicity from the beginning and retain the same features thereafter. PMID- 2984722 TI - Formation of cyclic adenosine monophosphate (cAMP) in the preovulatory rabbit follicle: role of prostaglandins and steroids. AB - The preovulatory increase in follicular prostaglandins (PG) stimulated by luteinizing hormone (LH) is dependent upon 3'-5'-cyclic adenosine monophosphate (cAMP) and is essential for ovulation. It has been proposed that follicular PG stimulate a second rise in cAMP, independent of LH. This study examined the temporal relationships among PGE2, PGF2 alpha 6-keto-PGF1 alpha, estradiol-17 beta, progesterone, testosterone, androstenedione and the biphasic increases of cAMP in follicles of rabbits. Does received indomethacin (IN, 20 mg/kg, i.v.; n = 30) or phosphate buffer (C; n = 30), 0.5 h before 50 ug of LH. At laparotomy at 0, 0.5, 1, 2, 4 or 8 h after LH, blood was collected from each ovarian vein and two follicles per ovary were aspirated of fluid and excised. Plasma and follicular tissue and fluid were assayed for PG and steroids. Tissue and fluid were assayed for cAMP. In C does, cAMP (pmol/follicle) in tissue increased from 11.3 at 0 h to 14.2 at 0.5 h, decreased at 1 h (5.4) and increased linearly through 8 h to 14.5. In IN-treated does, cAMP remained high from 0.5 (13.2) to 2 h (16.3), decreased at 4 h (7.9) then increased again by 8 h (15.5). Indomethacin decreased all PG in follicular tissue but 6-keto-PGF1 alpha rose after 2 h, whereas PGE2 and PGF2 alpha did not. Estradiol-17 beta, progesterone, and androstenedione did not vary with treatment; testosterone was increased (P less than .05) by IN. PGE2 or PGF2 alpha may terminate the first phase of cAMP production, rather than initiate the second phase. PMID- 2984723 TI - [Percutaneous transhepatic bile duct endoprosthesis. Choice of material, technic, clinical results]. AB - Eighty biliary endoprostheses were introduced by the transhepatic route in sixty patients with obstructive jaundice. Complication rate was 21% (no mortality). Average survival time is sixteen weeks (maximum 53 weeks). In most patients, a 12 F teflon endoprosthesis was introduced four to five days after the initial catheter drainage. Patients in poor condition had a primary implant of a 9 F endoprosthesis. A combined transhepatic/transoral implantation was carried out five times. Results have shown that implantation of a prosthesis is as effective as a surgical bypass operation for palliation of obstructive jaundice and that it is better than catheter drainage. In-vitro experiments have indicated that failure of a 12 F prosthesis due to encrustation may be expected in about 23 weeks. This is in line with the survival time of patients with carcinomas. PMID- 2984724 TI - [Computed tomographic control of the course of primary brain tumors following radiotherapy]. AB - Two hundred and seventy-eight CT scans are presented which were carried out on 105 patients following radiotherapy for primary cerebral tumours. Of these, 90 had surgery and 15 did not. Amongst 63 patients who showed abnormal enhancement before irradiation, enhancement was reduced in 32, increased in ten and remained the same in 17 following treatment. Of the 42 patients who did not show enhancement before irradiation, this was observed in 24 patients after treatment, but not in the remaining 18. Changes in the pattern of contrast enhancement was very variable in both groups. Other signs of tumour (compression of ventricles, mid-line displacement, ventricular dilatation) showed no differences in the groups. Changes in the central nervous system due to irradiation were observed in 25% of cases. PMID- 2984725 TI - [Computed tomographic diagnosis of liposarcoma]. AB - One hundred and eighty-two fat-containing soft tissue tumours have been found in 27 400 CT examinations. Amongst these there were 22 malignant liposarcomas. CT is unable to give a tissue diagnosis but, nevertheless, provides some important criteria which make it possible to differentiate benign lipomas from lipomatous tumours, particularly liposarcomas, which require surgical intervention. PMID- 2984726 TI - [Lumbar computed tomography in a bicolor mode]. AB - The interpretation of lumbar computer tomograms depends on the qualitative and quantitative assessment of the contents of the spinal canal and of its surrounding structures. This is a time-consuming process. A bi-colour presentation is described which provides simultaneously qualitative and quantitative information. It speeds up the process of interpretation and improves accuracy. PMID- 2984728 TI - [Plasma cell granuloma of the lung]. AB - Plasma cell granuloma is a rare, benign and usually solitary round tumour of the lung. Although uncommon, it is found relatively more frequently among children and adolescents. The lack of symptoms and absence of characteristic features make diagnosis difficult. In adults, the differentiation from bronchial carcinomas is particularly difficult because of their radiological similarity. Consequently, thoracotomy and excision of the tumour is the treatment of choice. PMID- 2984727 TI - [Computed tomographic and roentgenologic findings in bronchiolo-alveolar carcinoma of the lung]. AB - Eighteen patients with alveolar carcinomas (ten solitary, eight disseminated) were examined pre-operatively by CT and conventional radiography and the results were compared. The 'pleura fingers' and air bronchogram were shown by conventional tomography as often as by CT in the solitary cases. In the disseminated form, CT is superior by showing small foci in the opposite lung. PMID- 2984729 TI - [ECG-triggered NMR-tomography of the heart. Pathological findings]. AB - Initial systematic examinations of the heart by ECG-triggered NMR are reported. Twenty-three abnormal findings were observed amongst 46 patients examined. Special indications are the search for cardiac aneurysms, intracardiac thrombi and changes from pressure and volume stress. PMID- 2984730 TI - [Magnetic resonance tomography (MRT) in temporal lobe lesions. Study of patients with psychomotor seizures]. AB - A comparative study between magnetic resonance tomography (MRT) and CT was carried out in 16 patients with temporal lobe epilepsy. The MRT studies were performed on a 0.35 T Magnetom with T.1 modes in a coronal plane. MRT proved to the superior to CT. CT demonstrated a discreet temporal lobe lesion in three patients and MRT in four patients. In addition, unilateral atrophy of the temporal lobe was demonstrated by MRT in six cases; these could not be diagnosed by CT. The lack of artifacts near the skull base, the possibility of producing coronal sections and the excellent tissue differential of MRT provide the basis for improved diagnosis of lesions in the temporal lobes. PMID- 2984731 TI - [Metal-dense structures in the paranasal sinuses--a reliable indication of aspergillosis?]. AB - The significance of mycotic concretions of 'metal density' in the diagnosis of aspergillosis of the paranasal sinuses has been analysed in 47 patients with this diagnosis. The incidence, types, radiological appearances and chemical composition of these very dense calcifications are discussed. Demonstration of these concretions, if metal foreign bodies and displaced dental fillings are excluded, are evidence for a mycosis of the paranasal sinuses. Absence of these dense structures does not exclude aspergillosis and was present in our clinical material in only 50% of cases. PMID- 2984732 TI - [Potentialities of radiological diagnosis of injuries to the dorsal pelvic ring]. AB - Thirty-five patients with fractures of the anterior pelvis, in whom there was clinical suspicion of trauma to the posterior part of the pelvis, were examined by conventional radiology, bone scintigraphy and computed tomography, and the results were compared. CT provided the most information and made it possible to evaluate the soft tissues and organs of the pelvis. Bone scintigraphy produced the same number of abnormal findings as CT (sensitivity 100%), but was unable to differentiate between lesions of the sacro-iliac joints, fractures of the sacrum, combined injuries or degenerative processes. A positive bone scintigram is a reliable finding, with a high predictive value. A negative result excludes a fracture or other lesions of the dorsal portion of the pelvis completely. Conventional radiographs provided a correct diagnosis in only 37.1% as compared with CT. PMID- 2984733 TI - Roentgenographical detection of cocaine smuggling in the alimentary tract. AB - We report five cases of cocaine smuggling, by hiding multiple drug-filled foreign bodies in the colon, either after oral ingestion or by placing them in the colon per rectum. Several smuggling methods in the gastrointestinal (GI) tract and their radiological findings on abdominal plain film are presented. Computed tomography (CT) is found to be a more accurate method in the detection of the drug-filled bags, because of its improved contrast resolution and the absence of projections of overlapping structures on the transversal sections. Subsequently, CT is highly recommended in cases of negative or doubtful findings on conventional abdominal radiographs. In all cases conservative management was used and the drug-filled capsules and condoms passed spontaneously. No complications, such as intestinal obstruction by the bags of cocaine or intoxication by rupture of their wall were observed. PMID- 2984734 TI - [Echographic differential diagnosis of scrotal contents. A contribution to evaluation of the method]. AB - Ten patients were selected from a clinical material comprising 336 sonograms of the scrotal contents in order to discuss the differential diagnosis of testicular and extra-testicular conditions. The decision whether a tumour is testicular or extra-testicular does not usually pose a problem, but may be impossible for small peripheral tumours. The shape of the epididymis and vas deferens and the structure of the testes is of significance in the differentiation between torsion and epididymitis. Rare diseases--myxoliposarcoma, infiltrating urothelial carcinomas from the bladder growing along the vas, lymphangiomas--are considered in the differential diagnosis of space-occupying lesions. Lymphangiomas produce a typical appearance on ultrasound. The criteria for the diagnosis testicular trauma, rupture, parenchymal tears and haematocoeles are defined. Differentiating liquid from solid is rarely a problem. A testicular abscess in the presence of chronic orchitis can simulate a solid tumour. PMID- 2984735 TI - [Sonography of the thyroid gland with high-resolution real-time equipment]. AB - The results of sonography of thyroid glands in 131 patients were compared with the findings on palpation and from scintigrams. High resolution sonography is the most accurate method of examination for demonstrating changes in the thyroid. In 40% cases sonography demonstrated lesions additional to the single abnormality found clinically. In 10% cases a suspected thyroid abnormality on palpation proved to be due to extra thyroid disease. Evaluation of the number, size and localisation of space-occupying lesions by means of sonography is superior to all other kinds of investigation. It is, however, not possible to determine whether a lesion is benign. An attempt has been made to define the usefulness of thyroid sonography and its place during routine diagnosis. PMID- 2984736 TI - [Diagnosis and differential diagnosis of breast cancer using intravenous DSA]. AB - Intravenous DSA of the breast is a new, hardly invasive, method for demonstrating carcinomas of the breast and for differentiating these from benign lesions. Carcinomas show distinct tumour-staining during the capillary phase, although the criteria which are used on conventional angiography are not recognisable. Semi malignant tumours show feeble staining. Benign diseases such as fibro-adenomas and focal sclerosing adenosis are not demonstrated by I-V DSA. The indication for intravenous breast DSA is for the clarification of suspicious findings where less invasive methods, such as mammography and cytology, have failed to give a definitive diagnosis. PMID- 2984737 TI - [111In-oxine-labeled granulocytes in Crohn disease and ulcerative colitis. Labeling and study technics]. AB - 111In-oxine "pure" granulocyte scanning allows an exact localisation of infiltrated bowel in Crohn's disease and ulcerative colitis. Fecal excretion of labeled granulocytes yields a good parameter of disease activity. In comparison to 111In-oxine "mixed" leukocyte scanning, this method is more specific in assessing disease activity and more sensitive in localising diseased bowel. Radiation exposure to blood cells is not as high as in "mixed" leukocyte scanning. Pure granulocytes were isolated by discontinuous density gradient centrifugation of an isolated "mixed" leukocyte fraction. PMID- 2984738 TI - [1 year's experience with a 57-cm image intensifier in a universal roentgen work station]. AB - 3342 patients were examined by means of a 57 cm image intensifier during one year. The installation permits examination with the patient standing or lying. The resolution was measured with a lead grid under conditions resembling actual practice and were found to be similar to those obtained with angiographic equipment. Using a medium format technique for examining the gut and vascular system resulted in radiographic quality which was not significantly different from the use of a large format technique. For examination of the skeleton, radiographic quality, using the intensifier, was markedly poorer than that resulting from Bucky examinations. There was no diagnostic difference when carrying out lymphograms. The large intensifier window of 53 cm was particularly advantageous for DSA of the pelvic and lower limb vessels when compared with smaller intensifiers. Possible savings of film and contrast medium costs are discussed. PMID- 2984739 TI - [Experimental and clinical determination of renal size after intravenous and intra-arterial administration of ionic and non-ionic contrast media]. AB - A quantitative comparison of changes in renal size after the intravenous injection of contrast media showed no difference between ionic and non-ionic contrast medium. Contrast media of similar osmolarity (Metrizamide and Ioxaglate with 470 and 490 mosn/kg H2O) did show significant variations. Evidently properties of the contrast medium, other that their osmotic behaviour, are of significance (chemical structure, viscosity, iodine content, specific toxicity, and the effect on the permeability of biological membranes, calcium-binding activity, effect on coagulation). Changes in renal size could be reproduced by repeating the contrast injection at 24 to 48 hours. PMID- 2984741 TI - [Combination lytic-endoscopic therapy of common bile duct retained stones. Irrigation procedures using a Seldinger catheter system]. PMID- 2984742 TI - [CT findings in fresh pelvic vein thrombosis]. PMID- 2984743 TI - [Asymptomatic calcified thrombus of the inferior vena cava]. PMID- 2984740 TI - [Aneurysmatic pancreatic pseudocyst. Angiography and dynamic computed tomography]. PMID- 2984744 TI - Sterno-clavicular hyperostosis in a case with psoriasis and HLA-B27 associated arthropathy. PMID- 2984745 TI - [Familial proximal and distal symphalangism with carpal and tarsal fusions]. PMID- 2984746 TI - Adenomyosis uteri diagnosticised by hysterosalpingography. PMID- 2984747 TI - [Small intestine enema using a rectosigmoid catheter]. PMID- 2984748 TI - [Secondary hyperthyroidism in a patient with acute myocardial infarct as a possible cause of a false negative study with technetium pyrophosphate]. PMID- 2984749 TI - Effects of diethyldithiocarbamate and selected analogs on cadmium metabolism following chronic cadmium ingestion. AB - Effects of ip treatment with diethyldithiocarbamate (DDTC), 4 carboxamidopiperidine-N-dithiocarboxylate (CAP-N-DTC), and N-methyl-N dithiocarboxyglucamine (MDCG) on cadmium (Cd) levels in selected mouse organs and tissues were assessed after mice were offered deionized water containing 0.05 mg/ml of CdCl2 X 2.5 H2O, 10 mg/ml of sucrose, and 0.125 microCi/ml of carrier free 109-CdCl2 as the sole drinking fluid for 15 days. Only 0.31 +/- 0.01 % of the ingested Cd was absorbed. Data obtained following treatment were compared with those obtained earlier in similar studies following ip Cd injection. In contrast to its action when administered after ip Cd injection, DDTC enhanced hepatic Cd burdens in mice which received Cd po. DDTC did, however, reduce renal Cd levels markedly after Cd ingestion, while enhancing brain Cd levels. CAP-N-DTC and MDCG, which were shown earlier to have no effect on Cd levels in striated muscle following ip Cd administration, effected significant reduction of muscle Cd concentrations after Cd administration po, while also reducing hepatic and renal Cd levels significantly. It was concluded that certain dithiocarbamates effectively mobilize and promote excretion of Cd which has been absorbed from one of the natural portals of entry. PMID- 2984751 TI - [Alveolar cell carcinoma and its histogenesis: a review]. PMID- 2984752 TI - [A case of pulmonary hypertension associated with liver cirrhosis and hepatoma]. PMID- 2984750 TI - DNA binding, cardiac superoxide production and cytotoxicity of daunomycin analogs. AB - Pyridoanthraquinones are potent antibacterial agents especially against gram positive organisms. We tested two major biologic actions of these compounds: DNA intercalation and superoxide (O2-) production in sarcosomes. Using the bathochromic and hypochromic shifts induced by intercalation, followed by Scatchard analysis, we calculated dissociation constants and the number of binding sites per base pair for several analogues. We compared O2- production using cytochrome c reduction. Unsubstituted compounds do not bind to DNA or change its melting temperature (Tm). Placing a morpholino or piperidyl group at C 5 enhances the binding to DNA. The tetracyclic compounds were equipotent at producing O2- and were 20-fold more active than daunomycin. These compounds were unusual in their solid tumor cytotoxicity. PMID- 2984753 TI - Phrenic afferent input to the lateral medullary reticular formation of the cat. AB - The afferent inputs from phrenic nerve stimulation to the lateral reticular formation of the lower brain stem were studied in anesthetized spontaneously breathing cats. The activity of reticular neurons was recorded by means of extracellular tungsten microelectrodes. Electrical stimulation of the central end of the right phrenic nerve evoked excitatory or inhibitory responses in the lateral reticular nucleus (LRN), in the nucleus ambiguus (AMB) and in a region dorsal to the AMB of ipsi- and contralateral sides. Phrenic afferents belonging to the flexor reflex afferent group were involved in these responses. The discharge pattern of the respiratory related units (RRU) of the AMB were exceptionally affected by phrenic nerve stimulations. It is concluded that high threshold phrenic afferents relay in the LRN before projecting to the cerebellar cortex. The overlapping of respiratory and non-respiratory afferents in the reticular formation may participate to the adaptations of respiratory and somatomotor functions during specific behaviors. PMID- 2984754 TI - [Case of multiple spinal cord metastases]. PMID- 2984755 TI - [Clinico-pathological conference: a 61-year-old patient with thoracic suffusion, weight gain and ankle edema]. PMID- 2984756 TI - [A case from practice (33). Patient G.R., born 1-30-1910]. PMID- 2984757 TI - Initiation and maintenance of latent herpes simplex virus infections: the paradox of perpetual immobility and continuous movement. AB - During the acute phase of infection, herpes simplex virus (HSV) is taken up by nerve endings and travels, probably as a noninfectious nucleocapsid, toward the neurons of sensory ganglia. Infectious virus can be detected in ganglia for a limited period, after which the virus enters into a latent phase. It appears that synthesis of deoxyribonucleic acid is not required and that an early viral protein and at least one additional late virus gene product are involved in the establishment of latency. The distinction between a "static" and a "dynamic" form of latency depends on the ability to detect viral activities in neurons and on whether these observed activities are expressed continuously or intermittently. The development of recurrent lesions following virus reactivation is an occasional event and is controlled by inducing agents and the state of the organism. The maintenance of latency depends on the number of neurons that become latently infected after the primary episode, the number of neurons in which reactivation takes place, the fate of the neuron after virus reactivation, and the possibility of renewed neuronal infections after each recurrent episode. Exogenous reinfections may also contribute to the maintenance of latency since they can lead to latent infections in nearby or distantly located sensory ganglia. Multiple latent infections may result also from a single primary infection by dissemination of the virus to distantly located ganglia. PMID- 2984758 TI - Hypothesis on the molecular basis of nononcogenic retroviral diseases. AB - Oncogenic and nononcogenic retroviruses have similar structures and replicate by reverse transcription of their viral RNAs. The nononcogenic retroviruses are distinguished from the oncogenic retroviruses by virtue of the chronic inflammatory or degenerative disease they cause. These diseases include visna maedi of sheep, caprine arthritis and encephalitis, equine infectious anemia, and chronic spongiform polioencephalomyelopathy of mice. The basis for the particular disease caused by the nononcogenic retroviruses seems to be the selective infection of specific end-stage cells. A common factor in these slowly progressing diseases is lifelong persistence of virus with viral spread either in the face of host immunity-antigenic drift or by modulation of the host immune response. The molecular basis for antigenic drift in visna virus and its biologic significance are discussed. PMID- 2984759 TI - Equine infectious anemia virus: immunopathogenesis and persistence. AB - Equine infectious anemia (EIA) is a chronic, relapsing infectious disease of horses caused by a nononcogenic retrovirus. Virus persists in infected animals for life and can be reliably detected by serologic tests that measure levels of antibody to the major structural protein of the virus. Periodic virus replication in macrophages leads to an immunologically mediated acute disease characterized primarily by severe anemia. Recrudescence of acute EIA is the result of antigenic variation of the surface glycoprotein of EIA virus. The frequency and severity of clinical episodes of EIA decrease in most horses, leading to an inapparent carrier state. This cessation of clinical illness is probably brought about by the ability of the infected animals to eventually achieve a threshold efficiency of the immune response against antigenic epitopes common to all EIA virus strains. PMID- 2984760 TI - Retroviral spongiform polioencephalomyelopathy. AB - Aging wild mice (Mus musculus domesticus) from several different trapping areas in southern California are uniquely prone to a naturally occurring hind-leg paralytic disease and/or lymphoma. Both conditions are caused by an indigenous ecotropic murine leukemia virus (MuLV). These mice have a lifelong persistent viremia with total immunologic tolerance to the virus. The characteristic pathologic features are centered on the anterior-lateral horns of the lumbosacral spinal cord and consist primarily of a noninflammatory spongiform change, with reactive gliosis and neuronal dropout. The main cause of neuronal death apparently is abortive intracytoplasmic replication of virus particles. Genetic control of the naturally occurring disease in wild mice is achieved by segregation of a dominant ecotropic virus restriction gene, Akvr-1R/Fv-4R. The neurologic and/or neoplastic diseases are readily reproduced by experimental inoculation of newborn susceptible laboratory mice with purified, cloned ecotropic virus derived from the affected wild mice. The biologic and pathologic features of the experimental paralysis closely resemble those of the natural disease. This nononcogenic retroviral disease is useful in understanding the molecular basis of direct, virus-induced, neuronal and glial cell degeneration and their sequelae. PMID- 2984761 TI - [Early diagnosis of villonodular synovitis and synovial sarcoma of the ankle]. PMID- 2984762 TI - [Cylindroma on a pleomorphic adenoma. Apropos of 2 case reports]. AB - Among a group of 863 patients with epithelial tumors of the salivary glands, including 470 with pleomorphic adenomas, the latter were associated with carcinomas in 20 cases and of these two were adenoid cystic carcinoma. In both cases these were isolated parotid tumors, lacking functional symptomatology and failing to recur 2 and 4 years respectively after parotidectomy. Diagnosis was mainly by histology and was based on the finding of a co-existing pleomorphic adenoma and an authentic adenoid cystic carcinoma, not to be confused with the benign pseudo-cylindromatous appearance sometimes detected in simple pleomorphic adenomas. This association of adenoid cystic carcinoma and pleomorphic adenoma increases the risk of recurrence and of metastases--long-term follow up is necessary. Prognosis is dependent mainly on the quality of the initial surgical excision. PMID- 2984763 TI - [Chemodectoma. Apropos of a case located on the course of the inferior dental nerve]. AB - The case of a chemodectoma on the inferior dental nerve is reported. This site can be rarely found because no recent observation of this kind was published. PMID- 2984764 TI - Crystallography of supragingival and subgingival human dental calculus. AB - Selected area electron diffraction of sections and individual crystal fragments of human dental calculus has demonstrated that octacalcium phosphate, hydroxyapatite and whitlockite form the inorganic part of both supragingival and subgingival dental calculus. However, the major constituents in supragingival calculus are platelet-shaped crystals of octacalcium phosphate and needle-shaped crystals of hydroxyapatite, while bulk crystals of whitlockite is the predominant component in subgingival calculus. The subgingival samples seemed to be better crystallized than the supragingival ones. The results obtained by the electron optical and X-ray powder investigations are in good agreement. PMID- 2984765 TI - Molecular cloning and characterization of the cDNA coding for hepatitis B virus surface antigen. AB - Hepatitis B virus surface antigen (HBsAg) mRNA has been enriched from a hepatoma cell line (PLC/PRF/5) by specific polysome immunoprecipitation and used for cDNA cloning. A HBsAg cDNA clone was identified by in situ hybridization with a cloned viral probe. It was characterized by restriction endonuclease mapping and DNA sequence analysis. Molecular hybridization of PLC/PRF/5 cellular DNA and RNA to [32P]-labeled HBsAg cDNA revealed the integration of at least six copies of the hepatitis B virus (HBV) DNA into the host genome and expression of three DNA species containing HBsAg-specific sequences. The possible role of HBV in the oncogenesis of primary hepatocellular carcinoma is discussed. PMID- 2984766 TI - Acquired immune deficiency syndrome--an overview. PMID- 2984767 TI - Twenty-eight year old caucasian female with passage of mucoid material in urine. PMID- 2984768 TI - Influence of buffer capacity and pH-dependent rheological properties of respiratory mucus on health effects due to acidic pollution. AB - Respiratory mucus is found to be a barrier to the penetration of H+ ions into the surrounding tissues. These ions are captured and bound in the mucus, e.g. as H bond cross-linkages between the molecules. They increase its viscosity, which has a bearing on the mucociliary transport, airway resistance and gas exchange. The ability to bind the H+ ions is dependent on the buffer capacity and pH value of the mucus. Persons with acid-saturated mucus (some asthmatics and bronchitics) constitute a risk group, for whom effects can be more easily triggered from the surrounding tissues than in persons with mucus of higher pH. The latter primarily react only by changes of the mucus viscosity, which has a minimum near pH = 7.4. This, together with the mucus buffer capacity, appears to constitute the basis of the health effects reported from experimental studies. A simple test for evaluating the expected sensitivity of persons to acidic pollution is described. PMID- 2984769 TI - The polyphosphoinositides revisited. PMID- 2984771 TI - NIMH emphasizes the basics. PMID- 2984770 TI - Construction and recovery of viable retroviral genomes carrying a bacterial suppressor transfer RNA gene. AB - The integration of retroviral genomes into cellular DNA can induce mutations by altering the expression of nearby cellular genes and can serve to identify the gene affected. The construction of a retrovirus that stably carries a suppressor transfer RNA gene from Escherichia coli has allowed facile recovery of the viral genome in vectors marked with amber mutations. This virus can be used for rapid isolation of cellular sequences at the site of proviral insertion. PMID- 2984772 TI - Protection of mice against fatal herpes simplex type 2 infection by liposomes containing muramyl tripeptide. AB - Intravenous administration of liposomes containing muramyl tripeptide phosphatidylethanolamine, a lipophilic derivative of muramyl dipeptide that activates macrophages to a cytolytic state in situ, significantly protected mice against lethal challenge with herpes simplex virus type 2. These findings suggest that the systemic activation of macrophages by liposomes containing an immunomodulator can lead to prophylaxis of severe infections caused by herpesviruses. PMID- 2984773 TI - Bond order and charge localization in nucleoside phosphorothioates. AB - In the recent literature on nucleoside phosphorothioate anions the structural formulas show a double bond between phosphorus and sulfur and a single bond between phosphorus and oxygen with a negative charge localized on oxygen. However, a review of physical data on these compounds shows the reverse to be the case; that is, in phosphorothioate anions the P-S bond is a single bond with a negative charge localized on sulfur, while the P-O bond order for exocyclic and nonbridging oxygens is greater than 1, approaching 2 in O-alkyl phosphorothioate monoanions and O,O-dialkyl phosphorothioates. The P-O bond orders in phosphorothioate dianions and trianions approach 1 1/2 and 1 1/3, respectively, owing to delocalization of negative charge among two or three oxygens. These conclusions are based on bond lengths obtained from x-ray crystallographic data and electron diffraction, the magnitudes of the effects of 18O on the 31P-nuclear magnetic resonance chemical shifts of phosphorus in nucleoside [18O]phosphorothioates, the pH-dependence of 17O-NMR chemical shifts in [17O]phosphate and [17O]thiophosphate, the vibrational spectra of thiophosphate di- and trianions, and the pKa (dissociation constant) values for phosphoric and thiophosphoric acids. PMID- 2984774 TI - Characterization of envelope and core structural gene products of HTLV-III with sera from AIDS patients. AB - The envelope (env) and structural (gag) gene products of human T-cell leukemia (lymphotropic) virus type III were identified by immunoaffinity chromatography, immunoprecipitation, and two-dimensional oligopeptide mapping methods. The env gene specifies a glycosylated polypeptide with a molecular weight of 160,000 (gp160) that is processed to gp120 and smaller gene products. The gag gene specifies two polypeptides of 70,000 and 55,000 molecular weight (p70 and p55), both of which contain p24, the major structural protein of the mature virion. The techniques in this study can be used to define the extent of variability of the env gene product among different virus isolates and may identify the nature and patterns of the humoral immune response that lead to an immunologically protected state. PMID- 2984775 TI - Hormone receptors and the liver. PMID- 2984776 TI - Growth hormone, somatomedins, and the liver. PMID- 2984777 TI - Properties of ouabain-resistant variants of a polarized established cell line. PMID- 2984778 TI - Vitamin D-dependent calcium transport. AB - Vitamin D must be metabolized to 25-OH-D3 or calcidiol in the liver and subsequently to 1,25-(OH)2D3 (calcitriol) in the kidney to produce its physiological actions. Calcitriol stimulates calcium and phosphorus transport reactions in the small intestine and together with parathyroid hormone stimulates calcium transport reactions in bone and kidney. In the small intestine calcitriol brings about its response in a complex manner. It provides a very rapid response on existing villus cells, causing a rise in calcium transport within 6 h postadministration. This transport response subsides by 18 h, and a second response makes its appearance at 24-48 h. This second response is likely to be an action on crypt cells which then migrate into the villus region and bring about calcium transport. The molecular mechanism of calcium transport in the villus cells has been examined. Calcitriol localizes specifically in the nucleus within 0.5 h postadministration, and it does not localize in any of the other cell fractions. A receptor has been discovered for calcitriol, and interaction with this receptor is required for calcitriol to bring about its action in stimulating calcium transport. It is therefore believed that calcitriol, together with receptor, binds to specific portions of nuclear DNA to bring about transcription of specific genes that code for calcium and phosphorus transport proteins. Only one protein induced by calcitriol has been described. It is the calcium-binding protein found in the cytoplasm. Exactly how it functions in calcium transport is in debate, although the time sequence of its appearance appears to be consistent with a role in that system. It is likely that other gene products are induced by calcitriol, and they are currently being characterized. In the neonatal rat, active calcium transport does not make its appearance until 14-16 days postpartum. The intestine is not sensitive to vitamin D or calcitriol prior to this time. This lack of sensitivity is because of the lack of a receptor for calcitriol. The receptor to calcitriol can be precociously induced by hydrocortisone injections, or it can be delayed by adrenalectomy. Incubation of intestine from neonatal rat pups at 14 days postpartum with hydrocortisone brings about in vitro appearance of the receptor molecule. These results and results of studies involving genetic vitamin D resistance strongly suggest that calcitriol must function by interaction with the receptor, presumably by a nuclear mechanism, to bring about intestinal calcium transport. The molecular mechanism, however, remains to be described. PMID- 2984779 TI - Coordinated changes in potassium fluxes as early events in the differentiation of the human promyelocyte line HL-60. PMID- 2984780 TI - Ion fluxes and differentiation in transformed cell lines. PMID- 2984781 TI - Glucose and cation transport during in vitro adipocyte differentiation. PMID- 2984784 TI - Developmental and regulatory aspects of the sodium- and potassium-ion-stimulated ATPase in avian nerve and muscle. PMID- 2984782 TI - Regulation of Na+/H+ exchange in cultured human fibroblasts. PMID- 2984783 TI - Developmental aspects of sodium-dependent transport processes of preimplantation rabbit embryos. AB - It is well established that the trophectoderm of the preimplantation rabbit blastocyst can actively transport solutes that, in turn, are believed to mediate the accumulation of blastocoelic fluid. Consequently, the rabbit blastocyst serves as an ideal model to investigate epithelial transport mechanisms during embryonic development. The trophectoderm is considered to be an electrically "tight" epithelium, as values for the transepithelial electrical resistance and potential difference are high for both day-6 and day-7 p.c. blastocysts. However, during development the trophectoderm displays marked changes in epithelial transport properties, particularly between day 6 and day 7 p.c. These changes (see Fig. 14.1) are characterized by (1) the loss of a Na+-dependent methionine transport mechanism, (2) the development of an amiloride-sensitive component to Na+ influx, (3) a decrease in ouabain-sensitive oxygen consumption with a concomitant increase in ouabain binding, and (4) the acquisition of a furosemide sensitive NaCl cotransport system. During this period the rate of fluid accumulation doubles, resulting in a five-fold increase in blastocoele volume. We have also presented results showing that individual proteins of expanding blastocysts can be grouped with respect to their synthetic rates and that plasma membrane proteins can be uniquely characterized by their apical and basolateral domains. We suggest that the dramatic changes in epithelial transport mechanisms observed during preimplantation development of the rabbit blastocyst are a consequence of the changing developmental expression of trophectodermal membrane constituents. The blastocyst tissue is thus a useful model not only for understanding epithelial transport function in general, but also for understanding the physiological events associated with normal embryonic development. PMID- 2984785 TI - Regulation of turnover of Na+,K+-ATPase in cultured cells. PMID- 2984786 TI - Mineralocorticoid regulation of sodium and potassium transport by the cortical collecting duct. AB - Chronic high-dose mineralocorticoid hormone treatment of rabbits results in marked alterations of the structure and function of the cortical collecting duct. Most importantly, the reabsorption of Na+ and the secretion of K+ are increased. Intracellular microelectrode measurements provide evidence consistent with the idea that the increased transport of these ions is a result of changes in the membrane conductances and electrochemical driving forces for passive ion movement and of the activity of the Na+,K+-ATPase. Specifically, the Na+ and K+ conductances of the apical membrane are increased, and the cellular potential profile is altered to promote transcellular movement of K+ from the peritubular to the luminal fluid compartments. The associated increase in the activity of the Na+,K+-ATPase facilitates extrusion of Na+ from and accumulation of K+ into the cell. The resistance of both the apical and basolateral cell membranes are reduced with DOCA treatment, while the resistance of the paracellular pathway is increased. Consequently, the electrophysiological properties of the tubule epithelium reflect to a greater degree the properties of the transcellular pathway. PMID- 2984787 TI - Syndactyly reconstruction by a modified Cronin method. AB - In 11 patients (1 to 28 years of age) having elective surgical release of congenital or burn syndactyly in 1982-1983, we used the modified Cronin method of web reconstruction with double opposing, inverted V-shaped flaps and full thickness skin grafts. Postoperatively, the hands were splinted for three weeks, at which time we changed the dressing, with the patient under anesthesia. The hands were then immobilized for an additional three weeks. At the last clinic visit (13 to 20 months postoperatively) the hands were evaluated for adequacy of web space reconstruction, cosmesis, and postoperative complications. There was one partial loss of a full-thickness skin graft, which required regrafting when the patient prematurely removed his dressing. One proximal interphalangeal joint contracture required splinting. The web space reconstruction was judged adequate in all hands, and all patients and parents were pleased with the cosmetic results. This method of reconstruction gives good results and its web space reconstruction is technically easier than other methods. PMID- 2984788 TI - Cytological localization of adenosine kinase, nucleoside phosphorylase-1, and esterase-10 genes on mouse chromosome 14. AB - We have determined the regional locations on mouse chromosome 14 of the genes for mouse adenosine kinase (ADK), nucleoside phosphorylase-1 (NP-1), and esterase-10 (ES-10) by analysis of rearranged mouse chromosomes in gamma-irradiated Chinese hamster X mouse hybrid cell lines. Irradiated clones were screened for expression of the murine forms of these enzymes; segregant clones that expressed only one or two of the three markers were karyotyped. The patterns of enzyme expression in these segregants were correlated with the presence of rearranged chromosomes. The Adk gene was localized to bands A2 to B, Np-1 to bands B to C1, and Es-10 to bands D2 to E2. PMID- 2984789 TI - Replication of pSV2-gpt in COS-1 cells: stability of plasmid DNA in the presence and absence of biochemical selection. AB - We have previously demonstrated that COS-1 cell lines transformed by pSV2-gpt and maintained under biochemical selection replicate multiple copies of extrachromosomal plasmid DNA (1). We have now examined the replication and stability of this DNA in a representative cell line. In situ hybridization analyses revealed that intense replication of pSV2-gpt occurs in only a small subpopulation of cells and results from bursts of plasmid replication that occur periodically and spontaneously in the cell population. This suggests that COS-1 cells are only semipermissive for pSV2-gpt replication. No correlation was observed between levels of pSV2-gpt replication and the presence or absence of biochemical selection for the Gpt marker. However, growth of cells under nonselective conditions led to a rapid and progressive loss of pSV2-gpt DNA. This loss correlated with segregation of Gpt- revertants that lacked detectable plasmid sequences. Hence, maintenance of pSV2-gpt in the cell line was dependent on continuous biochemical selection. Stable replication of pSV2-gpt could be observed as late as four months after transfection, suggesting that this system might be useful for propagation of cloned DNA in COS-1 cells for extended periods of time. However, by nine months, extensive rearrangements of pSV2-gpt sequences were detected, indicating ultimate instability of the plasmid in the host cells. PMID- 2984790 TI - Chromosomal assignment of human sequences encoding arginine vasopressin neurophysin II and growth hormone releasing factor. AB - Complementary DNA clones encoding bovine vasopressin and human pancreas growth hormone releasing factor have been used to map homologous sequences in the human genome. Assignment of both cloned sequences to human chromosome 20 was accomplished by hybridization of insert DNAs to a panel of human-rodent somatic cell hybrids. Both these probes have been used to examine the structure of their respective genes in DNA from various individuals. No restriction fragment variants for growth hormone releasing factor have yet been found. Analysis of populations for restriction fragment length polymorphisms associated with disease states involving arginine vasopressin is underway. PMID- 2984791 TI - An indirect fluorescent antibody test for antibodies against HTLV-III. PMID- 2984792 TI - Successive urinary papillomavirus excretion, condylomata acuminata and urogenital epithelial malignant disease after renal transplantation. AB - Of 3 renal allograft recipients who were found to excrete papillomavirus in the urine, 1 has since developed a condyloma, 1 a condyloma and a keratinizing squamous carcinoma and 1 a transitional cell carcinoma. Peroxidase-antiperoxidase staining has confirmed the presence of papillomavirus antigen in the condylomas. The significance of these findings is discussed. PMID- 2984793 TI - Serum antibodies to human T-cell leukaemia virus type I in different ethnic groups and in non-human primates in South Africa. AB - The prevalence of humoral antibodies to human T-cell leukaemia virus type I (HTLV I) was investigated in different ethnic groups and in non-human primates in South Africa. Serum antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) using either disrupted whole HTLV-I or purified p24 core protein (p24 HTLV-I) as antigens. ELISA was complemented by direct radio immunoprecipitation assays using either purified iodinated p24 HTLV-I or radiolabelled lysates of an HTLV-producing cell line as antigen followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis of the immunoprecipitates, and by immunofluorescence using the HTLV-I-producing cell line HUT-102 as antigen. Antibodies were demonstrated in 3,5% of Asians, 3,5% of blacks and 4,1% of coloureds, but not in whites, and also in 29% of vervet monkeys and 33% of baboons. We conclude that HTLV-I or closely related viruses cause widespread infection in non-human primates in South Africa and in a lower percentage of humans, including apparently healthy blood donors. We are currently isolating retroviruses from seropositive reactors and investigating the possible relevance to disease in South Africa. PMID- 2984794 TI - Prevalence of HTLV-III antibodies in homosexual men in Johannesburg. PMID- 2984795 TI - Thiamine deficiency-induced gestational polyneuropathy and encephalopathy. A case report. AB - A 22-year-old multigravida presented with polyneuropathy and encephalopathy at 18 weeks' pregnancy. After excluding other applicable conditions, the diagnosis of a hyperemesis-induced thiamine deficiency was made. With the necessary vitamin supplementation the patient gradually recovered over a period of 4 months and was delivered of a normal infant at term. Gestational polyneuropathy and encephalopathy due to thiamine deficiency has very rarely been reported. The literature is reviewed with discussion of the differential diagnosis, the treatment and the prognosis. PMID- 2984796 TI - Reactivity of Bushman sera with HTLV-III--what does it mean? PMID- 2984797 TI - New technique for removable implantation of radionuclides in central nervous system neoplasm by ultrasonic guidance. AB - Introduced here is a new technique for the insertion of removable tubes into a brain tumor guided by a real-time ultrasonic B-mode scanner as a preliminary stage for interstitial brachytherapy of malignant brain tumors. Because tube insertion is conducted while watching in real time the tumor echo displayed on the cathode ray tube, the tube-positioning locations for insertion into the tumor can be freely selected with this technique. Additionally, the length of the tube that remains within the tumor bulk can be easily measured and recorded during the operation. Moreover, in that all these procedures can be carried out in the operating room, the operation will be performed under aseptic conditions and in a shorter time. These are the major advantages of this technique. PMID- 2984798 TI - Distribution of arterial blood flow in human hepatic cancer during chemotherapy- examination by short-lived 81mKr. AB - Using the characteristics of short-lived radioisotope, 81mKr (half-life = 13 seconds), we studied the distribution of arterial blood flow of human hepatic cancers during intra-hepatic arterial chemotherapy. Sixteen patients with primary and metastatic cancer were catheterized into the hepatic artery during operation. 81mKr solution was continuously infused via the hepatic artery with the patient in the supine, upright, right-side down, and left-side down positions. The frontal and right-side images of the liver were taken with a scinticamera. The tumor/nontumor ratio of the hepatic arterial flow in the supine position was more than one in 14 of 15 patients. The patterns of arterial flow distribution of 81mKr in the four positions were compared. In 11 patients the tumor/nontumor ratio of one of the other positions was significantly higher than that of the supine position. We expect that this quantitative measurement will be a good indicator for evaluation of a drug's accessibility to the tumor in the first pass. Moreover, the choice of the most effective position for intra-arterial infusion chemotherapy by this test will be possible to increase the relative amount of drug reaching the tumor. This technique may be applicable mainly to short-term drug infusion. PMID- 2984799 TI - [Outpatient polyclinic care in renal diseases]. PMID- 2984800 TI - Inhibition of human platelet aggregation by vitamin K. AB - The effect of several vitamin K (Vit K) analogues on the aggregation of human platelets was examined. The analogues were potent inhibitors of aggregation induced by ADP, thrombin, collagen and arachidonate but were less active against aggregation induced by the calcium ionophore A23187. Vit K3 also prevented platelet membrane phosphatide breakdown induced by collagen. These effects were not due to a direct inhibition of enzymes involved in the liberation of arachidonate or its subsequent transformation. The analogues exerted no effects on enzymes regulating intraplatelet cAMP. However, these effects could be overcome by increasing extracellular Ca++ levels, indicating a possible interaction with Ca++ regulation in platelets. PMID- 2984801 TI - An analysis of inhibitory effect of cyclic GMP on arachidonic acid- or collagen induced aggregation in rabbit platelets. AB - Aggregation of washed rabbit platelets induced by arachidonic acid (AA) or collagen was inhibited by nitroprusside (NP) and 8-bromo cyclic GMP in a concentration-dependent manner. Although NP and 8-bromo cyclic GMP inhibited the AA-induced aggregation, these agents did not affect the conversion of exogenous AA to PG endoperoxides and TXA2 (which were observed as TXB2). On the other hand, collagen caused release of AA from phospholipids and sequential formation of TXB2 in [14C]AA prelabeled platelets. In contrast with the case in which exogenous AA was used, NP and 8-bromo cyclic GMP inhibited the collagen-induced formation of TXB2 by preventing the liberation of endogenous AA. These results indicate that cyclic GMP has at least two different inhibitory actions in platelets; one is the inhibition of AA release from phospholipids and the other is the inhibition of the action of TXA2 in platelets. PMID- 2984802 TI - Effect of a fibrin(ogen)-derived vasoactive peptide on polymorphonuclear leukocyte emigration. AB - A vasoactive peptide known to increase vascular permeability and corresponding to residues 30-43 of the human fibrinogen B beta-chain induced polymorphonuclear leukocyte emigration in rabbit skin in vivo. The leukocyte emigration was much stronger after 2 h than after 0.5 h. Addition of prostaglandin E2 (PGE2) did not influence the chemotactic activity, which might possibly be explained by a known PGI2 releasing capacity of this peptide. PGE2 enhanced the leukocyte emigration due to leukotriene B4. PMID- 2984803 TI - Prevention of coronary platelet aggregation with a phosphodiesterase inhibitor RX RA 69. AB - The formation and prevention of coronary platelet thrombi (CPT) was studied in a modified Folts model in 23 anaesthetized dogs. The left circumflex coronary artery was acutely damaged and narrowed until resting flow started to fall. Spontaneous sharp decrease of flow indicated the acute formation of CPT. Intravenous infusion of 30 ng/kg/min of PGI2 prevented the formation of CTP. The effect lasted 3-7 min after termination of the infusion. RX-RA 69 a potent inhibitor of platelet phosphodiesterase (IC50 of 1 X 10(-9) mol/1) inhibited the formation of CPT for 9 and 18 min when 60 and 120 micrograms/kg were administered i.v. The results demonstrate that platelet aggregation induced by acute damage of the vascular wall can be inhibited by a potent PDE inhibitor. PMID- 2984804 TI - [Organic solvents. Damage to the peripheral nervous system]. PMID- 2984805 TI - [EHV1, to vaccinate or not?]. PMID- 2984806 TI - Toxaphene: accumulation in the adrenal cortex and effect on ACTH-stimulated corticosteroid synthesis in the rat. AB - Uptake and distribution of [14C]toxaphene was studied in the adrenals of rats using whole-body autoradiography. An accumulation of radioactivity was seen in the adrenal cortex (zona fasciculata) 1-24 h after a single gavage of [14C]toxaphene (16 mg/kg b.w.). In in vitro studies toxaphene was found to inhibit ACTH-stimulated corticosterone synthesis in the cultured rat adrenocortical cells (IC50 2.8 X 10(-5) M). Moderate but significant inhibition (P less than 0.001) of ACTH-stimulated corticosterone synthesis was also observed in the adrenocortical cells isolated from rats after a prolonged exposure (5 weeks) to low levels (1.2 ppm) of toxaphene in feed. The results indicate a direct adrenotoxic effect of toxaphene. PMID- 2984807 TI - Effect of alcohol intake on some disturbances induced by chronic exposure to carbon disulphide in rats. II. Biochemical and ultrastructural alterations in the peripheral nerves. AB - The effect of chronic ethanol (EtOH) administration on CS2-induced peripheral neuropathy in rats was studied. Rats were exposed to 0.8 mg/l CS2 for 12-15 months and to 10% EtOH as only drink during the last 4 months of the experiment. Some biochemical correlates of Wallerian degeneration of the peripheral nerve were estimated and ultrastructural examinations of the peripheral nerves were performed. It was shown that EtOH augmented the alterations in lipid content provoked by CS2 in the peripheral nerves, i.e. an increase in cholesterol esters and in the ratio of cholesterol esters to free cholesterol (E/F ratio) and a decrease in phospholipid content. The magnitude of ultrastructural changes induced by CS2 in the nerves was increased by EtOH. PMID- 2984808 TI - Behavioral effects of drugs as a function of maternal polybrominated biphenyl body burden. AB - Prior to breeding, female rats were dosed orally for 20 days with either 0.5 mg/kg polybrominated biphenyl (PBB), 5.0 mg/kg PBB, or the lecithin liposome vehicle. Male offspring of these 3 treatment groups did not differ with respect to the learning of an operant discrimination task. However, administration of phenobarbital or d-amphetamine impaired the behavior of the offspring of low-dose PBB dams less than that of the offspring of controls. The behavioral effects of the drugs were generally inversely related to levels of liver PBB and activities of liver aryl hydrocarbon hydroxylase (AHH). PMID- 2984809 TI - Extension of platelet concentrate storage by addition of sodium bicarbonate. AB - Viability of platelet concentrate (PC) stored in polyvinylchloride bags in an elliptical rotator at 22 degrees C (standard PC) was assessed by in vitro tests, and an alternate approach to extending the shelf-life of PC by the addition of hypertonic sodium bicarbonate (test PC) was investigated. The fall in the pH which occurred during storage in standard PC was arrested in test PC. Furthermore, platelets stored under these test conditions maintained their morphology better than in standard PC as judged by their mean platelet volume and platelet distribution width. Recovery of stored platelets from hypotonic shock at 37 degrees C following resuspension in fresh plasma was better for test platelets. Results indicated that platelets in standard PC were viable up to day 3 but were not viable at day 7. Platelets store better in PC to which sodium bicarbonate has been added and behave as viable platelets up to 7 days. PMID- 2984810 TI - Faster immunological recovery after bone marrow transplantation in patients without cytomegalovirus infection. AB - The following findings were noted among 45 bone marrow transplant recipients. The patients without cytomegalovirus (CMV) infection or chronic graft-versus-host disease (GVHD) showed normal lymphocyte stimulation in vitro by concanavalin A (Con A) more than 3 months after transplantation, and normal stimulation by phytohemagglutinin (PHA), anti-beta 2-microglobulin (A-beta 2m) and protein A (SpA) after 6 months. In contrast, the patients who had CMV infection without chronic GVHD had Con A and SpA responses within the normal range after 12 months and reduced lymphocyte responses to PHA and A-beta 2m more than 12 months after transplantation. The patients with chronic GVHD had reduced responses to all of these four mitogens after more than 12 months. In comparison with other patients those who later developed chronic GVHD showed an increased mixed lymphocyte culture stimulation during the first 3 months that decreased between 6-12 months. Patients with chronic GVHD still had reduced IgA levels at 12 months after transplantation. Patients with CMV infection, but without chronic GVHD, had higher percentages of lymphocytes with surface membrane Ig than healthy controls during the first 3 months after transplantation. The data suggest that CMV infection, regardless of chronic GVHD, delays immunologic recovery after marrow transplantation. PMID- 2984811 TI - Primary cytomegalovirus infection and its prevention after autologous bone marrow transplantation. PMID- 2984813 TI - Radionuclide scintigraphy in cytomegalovirus-related renal allograft dysfunction. PMID- 2984812 TI - Viral infections in kidney transplant recipients: lack of correlation with skin cancer. PMID- 2984814 TI - [Possible role of the T antigen in inducing chromosome aberrations in SV40 virus transformed cells]. AB - A possible role of the simian virus 40 T antigen in chromosome damages in transformed cells was examined. Two lines of Golden hamster embryonal fibroblasts, transformed by SV40 tsA30 and ts239 mutants (He30 and He239, respectively), were incubated at nonpermissive (40.5-41 degrees C) or permissive (33 degrees C) temperatures. Chromosome aberrations were registered in either subline after 3, 6, 9 and 12 weeks of cultivation under the above conditions. In the both cell lines kept at 33 degrees the frequency of aberrant metaphases and the number of chromosome breaks per cell increased drastically by week 3 of cultivation, and such a state was preserved up to week 12. The frequency of aberrant metaphases in cells cultivated at 41 degrees was maintained at the constant level (He239) or at slightly higher than that in the original culture (He30). The sublines He239, originally incubated at 33 or 40.5 degrees, were then shifted to 40.5 and 33 degrees, respectively. As a result the number of chromosome aberrations either decreased (33----40.5 degrees) or increased (40.5-- -33 degrees) as early as on day 2, and these patterns were stabilized at the level corresponding to the new conditions. We assayed the induction of DNA breaks in cells, grown at the permissive or nonpermissive temperatures, by using DNA sedimentation in the alkaline sucrose gradient. The DNA sedimentation peaks of cells cultured at 37 and 41 degrees coincided, whereas the DNA of cells cultured at 33 degrees was represented by shorter fragments. PMID- 2984815 TI - Pulmonary infection with Mycobacterium gordonae in the presence of bronchial carcinoma. PMID- 2984817 TI - [Undifferentiated small cell carcinoma of the renal pelvis with massive invasion of the kidney]. PMID- 2984816 TI - Polyamines and nucleic acids in gestational trophoblastic tumors. AB - To test the relationship between tumor malignancy and content and distribution of polyamines and nucleic acids, 2 forms of human gestational trophoblastic tumors were examined: the hydatidiform mole (self-limited form) and the human chorio carcinoma (invasive form) xenografted into nude mice. The results indicate that there are 2 significant differences between the choriocarcinoma and the mole: 1) the choriocarcinoma is characterized by increased polyamine and nucleic acid levels, 2) tissues differ in their putrescine:spermidine and spermidine:spermine ratios. There is an increase in polyamines in the urine of mole-bearing patients over that of normal controls. The correlation between putrescine and spermidine with the chorionic gonadotropin indicates that these 2 polyamines reflect the biological activity of the mole. PMID- 2984818 TI - [Granulocellular myoblastoma of the breast: the hazards of extemporaneous peroperative frozen section]. PMID- 2984819 TI - Rhabdoid tumor of the kidney: a case report. AB - Rhabdoid tumor of the kidney may simulate other pediatric renal tumors radiographically but it has a characteristic clinical course with a high incidence of intracranial tumors. This clinical course and its implications for prognosis and radiographic work-up are discussed. PMID- 2984820 TI - Intrarenal hemangiomas in the Klippel-Trenaunay syndrome. AB - The Klippel-Trenaunay syndrome is composed of the triad of unilateral limb hypertrophy, abnormalities of the deep venous system, and port-wine hemangiomas. An interesting case is presented in which there were multiple abnormalities of the renal veins and intrarenal hemangiomas resulting in renal failure in addition to the usual peripheral abnormalities. PMID- 2984821 TI - Spinal cord ischemia after ligation of both internal iliac arteries during radical cystoprostatectomy. AB - In radical cystectomy where control of the blood supply is desirable, ligation of both internal iliac arteries is to be avoided. The lumbar part of the spinal cord and cauda equina receive arterial radicular branches via the posterior trunks of the internal iliac arteries. Thus, bilateral ligation of these vessels might cause ischemia of the lower part of the spinal cord. PMID- 2984822 TI - Brachial plexus neuropathy from metastatic testicular seminoma. Prolonged survival after surgery and radiation therapy. AB - Prolonged survival and cure of brachial neuropathy were accomplished following subtotal removal and radiation therapy to metastatic seminoma involving the soft tissues of the right upper arm. Metastasis developed six years after orchiectomy for seminoma, and the patient is free of disease six years after treatment of metastatic disease. PMID- 2984823 TI - Nonepithelial neoplasms arising within vesical diverticula. AB - Two cases of rare vesical tumors, a paraganglioma and a malignant fibrous histiocytoma, are reported. The occurrence of these tumors within the urinary bladder is rare; their occurrence within bladder diverticula appears unique. The clinical, radiologic, and pathologic findings are summarized. PMID- 2984824 TI - Glomus tumor of glans penis. AB - Glomus tumor is an infrequent skin lesion and a rare lesion on the male genitalia. This is the second reported case of this lesion on the penis. PMID- 2984825 TI - Clinical evaluation of 99mTc-DMSA renogram. AB - Two hundred-two 99mTc-DMSA renograms for urologic problems were evaluated. Some technical aspects of the examination and the value of the scintigraphic depth estimation are discussed. Pre- and postoperative uptake values in patients with renal surgery and sequential postoperative examinations are considered. The value of DMSA renograms in predicting recovery in obstructive uropathy and in deciding to opt for conservative therapy or nephrectomy is discussed. PMID- 2984826 TI - [Epidemic outbreak of conjunctivitis due to the Coxsackie B1 enterovirus]. PMID- 2984827 TI - Hepatic neoplasia. AB - The clinicopathologic features of hepatic neoplasms such as hepatocellular adenomas, hepatocellular carcinomas, bile duct carcinomas, and hepatic carcinoids are presented. The authors also discuss the diagnostic and therapeutic approaches to hepatic neoplasia. PMID- 2984828 TI - Canine ovarian tumors of germ cell origin. AB - Fourteen ovarian germ cell tumors were diagnosed in the dog--seven dysgerminomas and seven teratomas. Six of the teratomas had immature components and were judged malignant; three had metastasized. One dysgerminoma had metastasized. The dysgerminomas developed in dogs ten years of age and older, and the teratomas developed in dogs nine years old and younger. PMID- 2984829 TI - Ultrastructural morphology of leukemic cells in the cat. AB - Transmission and scanning electron microscopic characteristics of leukemic cells from eight cats with spontaneous leukemia are described. Nuclear blebs, myelin figures, cytoplasmic microfibrils and C-type virus were seen more frequently in leukemic cells. Surface ridges and ruffles seen with the scanning electron microscopy were helpful in distinguishing myelogenous from lymphogenous leukemia. PMID- 2984830 TI - Dysentery caused by Escherichia coli (S102-9) in calves: natural and experimental disease. AB - A dysentery syndrome was recognized among the Institute's calves at 18 to 21 days of age. It was reproduced experimentally in gnotobiotic calves with an atypical Escherichia coli (S102-9) isolated from the affected calves. In both natural and experimental disease the calves passed copious bright red blood in the feces and developed diarrhea. Walls of the colon and rectum were thickened, and the mucosa was reddened and covered by an exudate that contained mucus and blood clots. Bacteria were seen closely adherent to the luminal surfaces of enterocytes, often in cup-shaped depressions or on cytoplasmic pedestals. Microvilli were distorted, disorientated or absent. There was exfoliation of infected enterocytes and a mild acute inflammation of the underlying lamina. In two of five calves with natural disease, the adherent bacteria did not stain by the immunoperoxidase method with antisera raised against E. coli (S102-9). This indicated that there was possibly more than one bacterial cause of the syndrome. Lesions in experimentally infected calves were indistinguishable from those produced by some E. coli which are enteropathogenic for man, rabbits, and pigs. PMID- 2984831 TI - Maedi-visna in Britain. PMID- 2984832 TI - Vaccination against equine influenza. PMID- 2984833 TI - Infectious disease incidence among horses in France, Ireland and the United Kingdom during 1984. AB - An outbreak of equine viral arteritis in the American state of Kentucky led to a temporary ban being imposed by France, Ireland and the UK on the importation of horses from the USA during 1984. Sporadic cases of influenza caused by the type 2 strain of the virus were confirmed in France throughout the year and cases of strangles in thoroughbred mares and foals were reported from all three countries. No cases of contagious equine metritis were confirmed among the thoroughbred breeding population, although a number of outbreaks of the abortion form of rhinopneumonitis occurred. A small number of clinical cases of equine infectious anaemia were confirmed in France. PMID- 2984834 TI - Foot-and-mouth disease virus subtype A22 epizootic in Assam. PMID- 2984835 TI - A single dilution enzyme-linked immunosorbent assay for the quantitative detection of antibodies to bovine herpesvirus type 1. AB - Three serological assays were compared for detection of antibodies to bovine herpes-virus type 1. These were virus neutralization (VN), enhanced complement fixation (CF) and enzyme-linked immunosorbent assay (ELISA). The ELISA was developed using an infected cell lysate antigen and purified virus and was optimized in relation to antigen and antisera dilutions. The CF assay was enhanced by the addition of bovine complement. These 3 assays were compared for detection of: specific virus antibody titers; sero-conversions; early antibody response in experimentally-infected cattle. Both ELISA end-point titers and single dilution values were found to be more sensitive than the CF or VN assays for specific antibody level quantitation. With a single dilution ELISA test procedure a correlation was obtained between ELISA values and VN titers. Using the single dilution ELISA test the assay also detected antibodies in experimentally-infected cattle before either the VN or CF assays, and agreed with the VN test in 35/38 seroconversions found by 4-fold or more VN changes between acute and convalescent paired sera from naturally-infected animals. The single dilution ELISA was a rapid and sensitive test for routine antibody detection in bovine sera. PMID- 2984836 TI - Detection of antibodies to bovid herpesvirus 4 by ELISA. AB - An enzyme linked immunosorbent assay for antibodies to bovid herpesvirus 4 was developed using antigen prepared by detergent lysis of infected cell cultures. The assay was used to study the immune responses of experimentally-immunised calves. The results correlated well with the indirect fluorescent antibody method. A viral neutralizing antibody response could not be demonstrated in the calves. PMID- 2984837 TI - Ovarian lesions in heifers exposed to infectious bovine rhinotracheitis virus by non-genital routes on the day after breeding. AB - Twelve heifers were exposed to either a Colorado infectious bovine rhinotracheitis (IBR) virus isolate or an Iowa IBR isolate obtained from a bovine respiratory disease outbreak. All inoculations were made on the day after the heifers had been in estrus and bred by an IBR virus-negative bull. Pairs of heifers were inoculated with each virus isolate intravenously, intramuscularly or exposed by aerosol. The heifers were killed 11-15 days after inoculation and their reproductive tracts and ovaries subjected to virological and pathological study. Virus was isolated from the ovaries of all 4 heifers inoculated intravenously and from 3 of the 4 heifers inoculated intramuscularly, but not from the ovaries of heifers exposed by aerosol. Virus isolations and lesions were, with only 1 exception, confined to the ovary containing the corpus luteum. In ovaries from which IBR virus was isolated, lesions in the corpus luteum ranged from focal necrosis and infiltration of mononuclear cells to diffuse hemorrhage and necrosis. Most of these ovaries also had necrotic follicles and a diffuse mononuclear cell accumulation in the stroma. Lesions were not found in ovaries from which IBR virus was not isolated. It was concluded that lesions are readily induced in the ovaries of post-estrus heifers as a result of hematogenous spread of IBR virus and suggest that the differences in lesion development observed with the 3 routes are related to whether or not a viremia occurred. PMID- 2984838 TI - Histological study of a case of recurrent olecranon bursitis with mixed calcium pyrophosphate dihydrate and apatite crystal deposits. AB - This is the report of a light and transmission electron microscopic study of an olecranon bursitis and of the adjacent distal tricipital tendon in an 83 year-old man. The data are compared with those of a similar study in the same patient performed 2 years ago. Calcium pyrophosphate dihydrate crystals were observed in the bursal fluid, in the inner part of the bursal wall (extracellular localization and intracellular phagocytosis) as well as in the peripheral part of the tendon. In addition, small apatite deposits were observed in the bursa and tendon by electron microscopy. The origin of these bursal deposits is discussed; it is suggested that they may be related to an exchange from the tendon to the remodelled bursal wall. PMID- 2984839 TI - [Beta-adrenergic receptors in healthy individuals and in patients with essential hypertension]. PMID- 2984841 TI - [Study of the alpha2-adrenoreactive system in man]. AB - A new approach for evaluation of the sensitivity of alpha 2-adrenoreactive system in men, based on main effects of clonidine, is developed. It may be used in investigation of both physiological and pathological processes in human body. PMID- 2984840 TI - [Effect of nonspecific hyposensitization (histamine and intal electrophoresis, intal electroaerosols) on cyclic nucleotide levels in children with respiratory allergy]. PMID- 2984842 TI - [Modification of the modification-restriction system in staphylococci]. AB - A new system of host specificity of DNA, called Sau67 according to the available nomenclature, was identified in Staphylococcus aureus 6782 strain by means of cross titration with staphylophage 729 considering that the phage exhibited the highly effective absorption properties. A total preparation of Sau67 methylases was isolated using ammonium sulfate fractionation. The enzyme preparation contained methylases of cytosine and adenine, where the activity of adenine methylases constituted only for 5% of the total methylase activity. As shown by kinetics of methylation low content of unspecific cellular nucleases was found in the St. aureus 6782 strain; these reasons are important for isolation of restricting endonucleases containing in the strain. 100 micrograms of protein of the total enzymatic fraction enabled to methylate the acceptory DNA at the maximal rate within 1.5 hr of incubation in phosphate buffer, pH 7.9. The fraction of cytosine methylases free of adenine methylating activity was obtained after chromatography on Sepharose blue with NaCl concentration stepwise gradient. PMID- 2984843 TI - [Effect of botulinum toxin on the activity of transport ATPases in biological membranes]. AB - Activity of transport ATPases was studied in erythrocyte membranes and synaptosomal fraction of cervical department of spinal cord obtained from rats in dynamics of botulinic C intoxication Na+, K+-ATPase was inhibited by the competitive type in the synaptosomal brain fraction at the preclinical period of intoxication and by the noncompetitive type at the step of skeletal muscle paresis. In erythrocyte membranes activity of Na+, K+-ATPase was inhibited by the mixed type at the preclinical period of intoxication and the enzymatic activity was inhibited by the noncompetitive type at the step of skeletal muscle paresis. The Na+, K+-ATPase from biological membranes was reactivated by unithiol and nicotinamide in dynamics of intoxication. The toxin was shown to activate Mg2+ ATPase in brain synaptosomal fraction. PMID- 2984844 TI - [Cyclic nucleotides in experimental uveitis]. AB - Content of cyclic nucleoside monophosphates was decreased in all the eye tissues in experimental toxico-allergic uveitis as well as penetration of cAMP into the fluid of anterior chamber of the eye. In various eye departments a dissimilar decrease of cyclic nucleotides occurred that had to be taken into consideration in a medicine prescription. PMID- 2984845 TI - [Effect of total ischemia and 3',5'-cAMP on the activity of the thermostable cytoplasmic inhibitor of Ca2+ ion transport in rat heart mitochondria]. AB - Activity of cytoplasmic inhibitor of Ca2+ transport in rat heart mitochondria was studied after total ischemia and incubation of heart homogenates with cAMP. Distinct inactivation of the inhibitor occurred under these conditions. The decrease of the inhibitor activity in ischemic myocardium appears to serve as a compensatory mechanism: 1. pyruvate dehydrogenase and the enzymes of tricarboxylic acid cycle were activated due to increase in Ca2+ concentration in mitochondria, 2. as a result of Ca2+ accumulation in mitochondria the elevated concentration of Ca2+ was decreased in myoplasm, which developed after impairment of plasmatic membranes and of sarcoplasmic reticulum membranes. PMID- 2984847 TI - [A rare cause of septic syndrome]. AB - The case report is presented of a 18-year old patient, who was admitted to the Haematology Department of the Hanusch Hospital with septic fever, an enlarged spleen and suspected bone marrow failure. Since the patient reported a stay in Sicily prior to the onset of his disease, an infection with Leishmania was suspected. The serological test was highly positive and Leishmania was also isolated from the spleen aspirate. Chemotherapy with Pentostam was successful and the patient made an uneventful recovery. This paper deals with the epidemiology of the disease and discusses the diagnostic approaches. PMID- 2984846 TI - Failure of passively administered anti-Rh to prevent secondary Rh responses. AB - Rh-negative women, immunized to Rh by previous pregnancies, with only low concentrations of IgG anti-Rh(D) in their plasma were assigned at random to test and control groups (7 subjects in each group). Both groups were challenged with an intravenous injection of 0.28 ml of Rh-positive red cells; in addition, the test group received 500 micrograms anti-Rh intramuscularly. 2 weeks after the injections, all subjects showed an increase in plasma anti-Rh concentration; levels in test and control groups were similar. It is concluded that in Rh immunized subjects with low levels of IgG anti-Rh a secondary response to Rh cannot be prevented by giving passively administered anti-Rh with the red cells. PMID- 2984848 TI - [Immunologic phenotypes of germinal center cell tumors]. AB - A panel of monoclonal antibodies applied to frozen sections of non-Hodgkin's lymphomas was used to establish clear-cut differences among the different entities of malignant lymphomas of germinal centre cell origin. 51 cases (18 centrocytic, 25 centroblastic-centrocytic and 8 centroblastic lymphomas) were included in this study. A clear-cut difference in the expression of the T65 antigen (Leu 1+) and the common acute lymphoblastic leukaemia antigen (CALLA) was found. Thus, centrocytic lymphomas predominantly expressed Leu 1, but not CALLA, whereas centroblastic-centrocytic lymphomas were always positive for CALLA, but not for the T65 antigen. Centroblastic lymphomas are virtually never positive with respect to either antibody. These findings suggest that, perhaps, two different phenotypes of centrocyte exist in centrocytic and centroblastic centrocytic lymphomas. PMID- 2984849 TI - [Serum angiotensin convertase activity and clinical course of sarcoidosis. Course studies in Lofgren syndrome]. AB - Serial measurements of SACE were performed in 24 patients with Lofgren's syndrome during a period of 32,4 +/- 8,0 months. Initially only 21% (5 patients) had elevated SACE, but the percentage increased during the first weeks of disease reaching 46% (11 patients). After regression SACE activities were in the normal range. In two patients (8%) 18 and 21 months, respectively, after onset of disease pathologically elevated SACE values were measured. Clinically relapses were observed. Our observations demonstrate that serial measurements of SACE are suitable for controlling patients with Lofgren's syndrome, indicating relapses and change to chronic course of sarcoidosis. PMID- 2984850 TI - [Nosocomial virus infections]. PMID- 2984851 TI - [Stress and the endogenous opioid system. I. Physiology and pharmacology of opioid peptides]. AB - In this article is given a survey on physiology and pharmacology of the opioid system. Opioid peptides are naturally occurring morphine-like acting metabolites of glycoprotein precursors. Of the opioid peptides proved hitherto in the organism met-, leu-enkephalin, dynorphin and beta-endorphin are characterized more in detail. Opioids react directly with opioid receptors. The opioid receptors are not a homogeneous system. Their distribution is depending on organs and species. Opioid peptides and receptors were proved within as well as outside the central nervous system. The main quantity of the endogenous beta-endorphin is stored in the pituitary gland. High concentrations of met-, leu-enkephalin and dynorphin are present in the gastrointestinal tract and in the adrenal glands. Opioid peptides are inactivated by enzymatic hydrolysis, in which case the splitting of the N-terminal tyrosine is decisive. The inactivation may be performed by amino peptidases, peptidyl dipeptide hydrolases or the angiotensin converting enzyme. The effect of the opioid peptides can be inhibited by opioid antagonists (naloxone, naloxazone). Up to now there are contradictory findings as to the presence of an endogenous opioid antagonist. In general, the presence of different opioid peptides and their different receptor preference indicate multiple functions in the organism. However, their physiological function is up to now only little clarified. PMID- 2984852 TI - [Malignant transformation of cutaneous cylindromas. 2 case reports and a review of the literature]. AB - We report on two female patients (age 78 and 81) suffering from multiple hereditary cylindromas which had grown, changed in color towards a bluish note, and partly exulcerated. The lesions were widely excised; plastic surgical reconstruction resulted in satisfactory healing. Histological examination showed similar features in both patients: We observed decreased differentiation of structures otherwise typical for cylindromas, polymorphism of cells and nuclei, mitoses, and beginning infiltration of the galea. Although there were no metastases found, malignant transformation was deduced from sudden growth, exulceration, as well as dedifferentiation of histological structures in combination with infiltrative expansion. PMID- 2984853 TI - [Importance of immunohistochemistry of skin tumors]. AB - Immunohistological demonstration of cell- and tissue-specific antigens in microscopic slides has remarkably influenced the classification of certain cutaneous neoplasms. The following markers are of particular interest in dermatopathology: cell surface antigens of lymphocyte subpopulations, intracytoplasmic immunoglobulins, intermediate filaments, neural markers, epithelial cell membrane antigens, and endothelial products. During the last years, these methods have contributed to our understanding of histogenesis and classification especially of cutaneous lymphomas, epithelial neoplasms, neurogenic tumors, and angiosarcomas. PMID- 2984854 TI - [Syringocystadenoma papilliferum as a differential diagnosis of a carcinoma of the lower lip]. AB - A 56-year-old patient showed a skin-colored tumor of the lower lip which had been photographically documented since his 10th year of age. On account of the localisation, increase of size during the last few months, as well as irregular epithelial hyperplasia observed in a punch biopsy from the periphery, we suspected the tumor to be a squamous cell carcinoma. Histological examination, however, revealed syringocystadenoma papilliferum. PMID- 2984855 TI - Organization and state of methylation of endogenous type C retroviral sequences in 129 mouse differentiated and undifferentiated teratocarcinoma cell lines. AB - Attempts to activate type C endogenous viruses in 129 mouse fibroblasts and in teratocarcinoma-derived cell lines have never been successful, although the genome of these cells contains xenotropic virus-related sequences. We have investigated the arrangement of these sequences and their methylation state by DNA restriction endonuclease digestion, electrophoresis of digests in agarose gels, Southern blotting and hybridization with specific probes. Our results show that the majority of the sequences are organized into two complete provirus families integrated at multiple sites in the cell genome and that they are hypermethylated in embryonal carcinoma cells as compared with differentiated cells. Having previously found a higher expression of viral RNA in 129 derived embryonal carcinoma cells, our data indicate an apparent direct correlation between methylation and type C virogenes expression. PMID- 2984856 TI - Comparison of structural and nonstructural proteins of virulent and less virulent Theiler's virus isolates using two-dimensional gel electrophoresis. AB - The Theiler's murine encephalomyelitis viruses (TMEV) are important neurotropic picornaviruses because they persist in the central nervous system (CNS) and produce an inflammatory demyelinating disease in the mouse, their natural host. Insight into the pathogenesis of this disease may come from studying the genetic and biochemical compositions of these viruses; therefore, in this report, the structural and nonstructural proteins specified by both highly and less virulent TMEV were examined. Using two-dimensional gel electrophoresis, structural and nonstructural proteins, originating from each of the three regions of the picornavirus genome (Kitamura et al., 1981; Rueckert and Wimmer, 1984), from nine TMEV isolates were compared on the basis of isoelectric points (pI). Proteins of two virulent TMEV (GDVII and FA viruses) had almost indistinguishable pI values, whereas two of the three major capsid proteins of the less virulent TMEV varied considerably. For example, the structural proteins VP1 and VP3 from seven less virulent viruses ranged from pI 6.3 to 6.9 and 6.5 to 8.3, respectively. On the other hand, the pI values of VP2 and nonstructural proteins from the less virulent TMEV varied relatively little. In general, structural proteins of each TMEV group had pI ranges unique to their respective biological group, while most nonstructural proteins were similar for all TMEV. The virus-specified proteins of Vilyuisk virus, which is serologically related to the TMEV and a possible cause of encephalomyelitis in man, had pI values similar to the less virulent TMEV. Finally, VP3 not only showed the greatest variation in pI among the less virulent TMEV, but it also was preferentially radioiodinated in intact virus from each of the two biological groups using the lactoperoxidase technique. PMID- 2984857 TI - [Innate forms of behavior and energy metabolism in the brain of the rat after phenazepam administration in the presence of inescapable painful stimulation]. AB - Initially low level of motor search activity in test situations ("open field", maze) in rats previously grouped according to their passive behaviour with a partner-victim (Simonov method), does not essentially change after 3-week administration of phenazepam; the activity level of cytochrome oxidase in the cortex and hypothalamus is lowered. Phenazepam administration in conditions of prolonged unavoidable painful stimulation brings to a sharp increase of motor search activity and aggressive behaviour. Simultaneously the cytochrome oxidase activity increases in the same way as during a stress without phenazepam. This fact points to the independence of behavioural and biochemical effects in this case. PMID- 2984858 TI - [Activity of callosal neurons of the visual cortex in the cat]. AB - Activity of 28 identified neurones of the visual cortex was recorded in cats immobilized by d-tubocurarine. Stimulation of the callosal body with a single stimulus or high-frequency train elicited a short-latency antidromic reaction of neurones in the visual cortex whose axons constitute the main part of the large cerebral commissure. Some commissural neurones responded to a single callosal stimulation by two action potentials the first one being antidromic, the second one being of long-latency postsynaptic origin. The second action potential was generated as a result of activation of axonal collaterals of the same neurone or the neighboring callosal neurones. More than a half of callosal neurones responded to a single stimulation of the lateral geniculate body by short-latency antidromic discharges and by long-latency postsynaptic reactions. These data indicate the existence of the systems of two-way neuronal connections, i.e. calloso-geniculate and geniculo-callosal ones. PMID- 2984859 TI - [Locoregional recurrence following the operative treatment of rectal cancer. Basic principles of prevention and therapy]. AB - The frequency of local recurrences has been observed in dependence on sex, tumour specific parameters (typing, grading, staging, macroscopical form of growth, level of primary tumour) and technical parameters (method of operation, distal margin of resection). In case of early diagnosis, there is a possibility of differentiating technically caused local recurrences from those caused by the tumour because of their localization. Recurrences caused by the tumour may proceed from incompletely removed primary tumours. Technically caused recurrences may be primarily due to insufficiently resected tumours, which could have been removed completely. Only technically caused recurrences make a curative second operation possible. If the criteria of radicality are strictly observed during the primary operation, there should be no need for the so-called 'surgically curative' local recurrences to be treated anymore. PMID- 2984860 TI - [Etiology of colon cancer--current aspects]. AB - Multiple factors such as tumour initiating, tumour promoting, tumour inhibiting are known to be involved in the aetiological process of most colon tumours. Dominant factors are associated with life style habits. Yet in spite of many experimental and epidemiological data it has not been possible, hitherto to give convincing recommendations for the primary prevention of this type of tumours. PMID- 2984861 TI - [General conservative operation in malignant ovarian tumors of the stage Ia (T1aN0M0)]. AB - In the Gynecological Clinic of the Medical Academy Dresden 585 women with primary malignant ovarian tumors were attended from 1953 until 1979. 118 patients (20,2%) were classified as stage I, 97 (82,2%) of them as stage Ia. A unilateral adnexectomy was carried out in only 25 women (25,8%) at stage Ia, 13 (13,4%) of them were younger than 40 and did not undergo additional therapy because they wanted to have a child. Six women were not subjected to additional treatment due to old age and general condition. Five of the 13 younger women gave birth to viable children after an average of 50 months, one patient was confined for the second time after further 24 months. Eighteen (95%) of 19 exclusively conservative operated on women survived the 5-year-period free of tumor. All these women were younger than 40. -After an average of 73 months six young women had to undergo relaparotomy and radical operation under suspicion of recurrence. In two cases a tumor recidivation was found, one where of was precancerous. Benefits and risks of the conservative approach in primary treatment are discussed and attention is drawn to the necessity of secondary radical operation. PMID- 2984862 TI - [Electron microscopic detection of viruses in cervix papilloma]. AB - The authors describe the electron microscope findings in a case of papilloma of the uterine cervix. In the nuclei of single koilocytes spherical inclusions measuring 47-56 nm were found. The ultrastructural pattern of the particles is entirely identical with one of the human papilloma virus particles. PMID- 2984863 TI - [Use of magnetotherapy in clinical neurology (review)]. PMID- 2984864 TI - The chemical synthesis and antiviral properties of an acyclovir-phospholipid conjugate. AB - The synthesis of acyclovir-phospholipid conjugate (2) is reported through an unambiguous one-step preparation of L-alpha-dimyristoyl phosphatidic acid triethylammonium salt (5). The biological activity of 2 as an antiviral drug has also been investigated. PMID- 2984865 TI - Abrasive bronchial brushing cytology. A preliminary study of 200 specimens for the diagnosis of neoplastic and nonneoplastic bronchopulmonary lesions. AB - Two hundred subjects with chronic respiratory symptoms with a suspicion of malignancy were selected for bronchial brushing cytology. Prior sputum examination had shown malignant squamous cells in two cases only. The cytologic appearances of the brushing smears were divided into five categories: 41 (20.5%) smears with positively malignant cells; 20 (10%) smears predominantly showing chronic inflammatory features; 31 (15.5%) smears with mainly acute inflammatory changes; 60 (30%) smears with normal cytologic features; and 48 (24%) smears unsatisfactory for cytologic interpretation. Thirteen patients with a positive cytology had a positive tissue biopsy for malignancy. Among the group with chronic inflammatory changes, acid-fast bacilli were identified in nine cases, and one smear showed frank tuberculous granuloma. In the unsatisfactory group, two cases showed malignant cells in the postbrushing sputum. There was one false negative report for malignancy in the entire study. This study confirms the sensitivity and accuracy of bronchial brushing cytology in the diagnosis of various bronchopulmonary lesions, especially malignancy and pulmonary tuberculosis, in India. PMID- 2984866 TI - Signet-ring-cell (colloid) carcinoma of the urinary bladder. Cytologic, histologic and ultrastructural findings in one case. AB - A 65-year-old female presented with a one-and-a-half-year history of intermittent, painless, gross hematuria. She was found to have a large bladder neoplasm. A cytologic diagnosis of signet-ring-cell (colloid) carcinoma was made on a bladder washing and was confirmed histologically by a cystoscopic biopsy. Subsequently, a radical cystectomy was performed. The cytologic findings of this rare pure signet-ring-cell carcinoma of the bladder, not previously described, are presented along with the histologic and ultrastructural findings. PMID- 2984867 TI - Adenoma malignum/urothelial-cell viral infections. PMID- 2984868 TI - Usefulness of bile cytology in the diagnostic management of patients with biliary tract obstruction. AB - Eighteen patients with evidence of biliary tract obstruction had a total of 29 satisfactory bile samples submitted for diagnostic cytology during a two-year period. These 29 specimens were reviewed in order to determine if bile cytology is useful in the diagnostic management of patients with obstructive biliary tract disease. Twenty-one of the bile specimens were from patients with malignant biliary stricture, and eight were from patients with benign biliary obstruction. Bile cytology was positive for carcinoma in eight samples from patients with malignant stricture and was inconclusive for malignancy in two. There were no false positives. The diagnostic specificity of bile cytology was 100%, the diagnostic sensitivity was 48%, and the diagnostic accuracy was 62%. When carefully collected and promptly processed, bile proved an excellent specimen for cytologic evaluation and was a valuable adjunct to other diagnostic procedures for the detection of carcinoma causing biliary tract obstruction. PMID- 2984869 TI - Effect of pretreatment of long-term ovariectomized rats with an LRH antagonist on LH release in vitro by LRH, elevated K+ or N6-monobutyryl adenosine 3',5' monophosphate (mbcAMP) plus theophylline. AB - The effect of the LRH antagonist (Ac-D-p-Cl-Phe1,2,D-Trp3,D-Phe6-D-Ala10)LRH (Org 30093) on pituitary LH release was studied, using pituitary glands of ovariectomized rats. In vitro, the antagonist had no detectable agonist activity in the concentration used, had no effect on the maximal LH release which can be induced by LRH and shifted the dose-response line of LRH to the right, without changing its slope. By this the antagonist fulfilled the conditions of purely competitive antagonist. Also, when present in vitro, the antagonist had no effect on LH release induced by raised K+, whether alone or in combination with mbcAMP plus theophylline. A single injection of Org 30093 decreased serum LH without inducing a change of pituitary LH content measured 24 h later. Twenty-four h after the sc injection of the agonist, LH release in vitro induced by LRH was affected differently, depending on the concentration of LRH used: the effect of low concentrations of LRH was inhibited, whereas the effect of high concentrations of LRH was augmented. Pretreatment with the agonist had no effect on LH release by raised K+ combined with mbcAMP plus theophylline, but slightly increased LH release by raised K+. PMID- 2984870 TI - Forskolin stimulation of naphthylamidase in guinea pig thyroid sections detected with a cytochemical bioassay. AB - Forskolin, from the roots of the Indian medicinal plant Coleus forskohlii, has recently been shown to be a potent stimulator of adenylate cyclase in many systems, including endocrine tissues such as the thyroid gland. We describe forskolin activation of beta-naphthylamidase activity in guinea pig thyroid tissue using the cytochemical bioassay (CBA) for thyroid stimulators. This CBA is the most sensitive bioassay for TSH and LATS-B currently available, being able to detect stimulation by doses as low as 10(-5) mU TSH/l and 10(-9) mU LATS-B/l. The dose-response curve to forskolin was bell-shaped (as is seen with TSH and LATS-B) with the ascending limb of the curve produced by 10(-13) M to 10(-12) M forskolin after a 3 min exposure time. Maximal stimulation was observed with 10(-12) M forskolin. However, the dose-response curve to forskolin was not parallel to that given by TSH, the slope of the ascending limb being much greater. It has been suggested that stimulation of beta-naphthylamidase activity in the CBA is via cAMP. We report that dibutyryl cAMP at doses from 10(-16) M to 10(-11) M produces a bell-shaped dose-response curve with a very broad peak response, again not parallel to that produced by TSH. Forskolin activation of beta-naphthylamidase in the CBA is unaffected by a 1:10(6) dilution of 11E8, a monoclonal antibody raised against solubilised TSH receptors, which binds to the TSH receptor and inhibits TSH stimulation. Although the precise location of forskolin action is not known, this is further evidence that forskolin acts at a post-surface receptor site. PMID- 2984871 TI - Physiological dosing of exogenous ACTH. AB - We evaluated the ACTH and cortisol responses to several doses of exogenous ACTH, and compared these values to the physiologic responses obtained in normal subjects undergoing insulin-induced hypoglycaemia. We determined that a cosyntrophin dose of 0.2 micrograms/kg body weight produced both ACTH and cortisol levels indistinguishable from the 'physiologic' stress-induced values. Since this dose is approximately 4 per cent of the standard 250 micrograms dose employed in tests of adrenal function, our findings suggest a need for caution in the interpretation of such tests. PMID- 2984872 TI - Demonstration of hydroxysteroid dehydrogenases and testosterone in the Sertoli Leydig cell tumor (androblastoma) tissue of the human ovary: an enzyme histochemical and immunohistochemical study. AB - Gynecological, endocrinological and histological tests on a 19-year-old female patient led to the diagnosis of Sertoli-Leydig cell tumor (arrhenoblastoma) of intermediate differentiation. For enzyme histochemical purposes the tumor tissue, removed from the right ovary by laparatomy, was frozen in liquid nitrogen. The following enzymes were demonstrated: nonspecific esterases, 3 beta-hydroxysteroid dehydrogenase (HSDH), 17 beta-HSDH, 11 beta-HSDH, and NADH tetrazolium reductase. Cryostat sections, prefixed with formaldehyde vapors, were used to localize testosterone production immunohistochemically with the PAP method. A large number of pseudotubules with Sertoli cells were observed; the Leydig cells in the interstitial space were often arranged in the form of islands. Strong nonspecific esterase activity weak 3 beta-HSDH activity, moderate 17 beta-HSDH activity, and strong 11 beta-HSDH activity were observed largely in the Leydig cells. Testosterone synthesis, demonstrated immunohistochemically, took place predominantly in the Leydig cells, but also to a small extent in the Sertoli cells. PMID- 2984873 TI - A high-affinity folate binding protein in human cerebrospinal fluid. AB - Binding of 3H-folic acid to a protein in human cerebrospinal fluid (CSF) was studied in equilibrium dialysis experiments at 37 degrees C and pH 7.4. The binding was of a high-affinity type (10(10) l/mol), and the relative molecular mass of the protein was 25,000 as determined by gel filtration. The concentration of folate binding protein in CSF, expressed as maximum folate bound per 1, was 0.3 nmol/l. This value was of the same order of magnitude as that of serum. PMID- 2984874 TI - Short latency somatosensory evoked potentials in Charcot-Marie-Tooth disease. A family with an intermediate form. AB - 9 patients with Charcot-Marie-Tooth Disease (CMTD) of intermediate type (PMA type II, 10), all from the same family, presented with a significant increase of the interpeak N9-N13 latency. This increase is already present in the pre-symptomatic phase of the disease and there is no significant difference between the various patients of different ages and clinical severity, indicating that the lesions appears very early and tends to establish itself equally early. Similar behaviour is also seen in the distal conduction velocity along the sensitive fibres, while the more proximal areas seem to be relatively spared. The authors interpret these data as an expression of a distal central peripheral sensory neuropathy. In contrast, the lesion of the peripheral motor fibres, particularly in the legs, has a different and more severe pattern of evolution. Alterations in central conduction time (N13-N20) were not seen in any of the 9 patients studied. PMID- 2984875 TI - Effect of natural estrogen on the serum level of follicle-stimulating hormone (FSH), estradiol and estrone in post-menopausal women and its effect on endometrium. AB - Serum FSH, estradiol and estrone levels were assayed in 35 post-menopausal women who received the three commonly available estrogen preparations Premarin, Progynova and Harmogen. The results were compared with the levels during treatment with Dixarit. Hormone levels did not change when Dixarit was used. The levels of estrone and estradiol increased significantly (p less than 0.01) when estrogen preparations were used. There was no abnormality in the endometrium at the end of the treatment period. PMID- 2984876 TI - Viral infections in pregnancy. PMID- 2984877 TI - The pattern of distribution of laminin in neurogenic tumors, granular cell tumors, and nevi of the oral mucosa. AB - Oral tumors of presumably neuroectodermal origin were stained with anti-laminin antibody by a double layered immunofluorescence technique. A marked positive staining for laminin was found in neurofibromas and neurilemmomas although the pattern of laminin distribution was slightly different. Accentuated staining was seen in Verocay bodies. In granular cell myoblastomas (GCM), small groups of tumor cells were encircled by laminin-positive material, whereas individual tumor cells were unstained. In nevi, diffusely spread nevus cells were surrounded by a rim of laminin, whereas when arranged in nests whole groups of cells were encircled by laminin as seen in the GCM. Ordinary oral fibromas included as controls were negative except for the expected positive staining of basement membranes normally occurring in the tissues. Immunohistochemical demonstration of laminin seems to be a valuable aid in differential diagnosis of soft tissue tumors and may provide useful information about the pathogenesis of various lesions. PMID- 2984878 TI - Invasiveness of Salmonella typhimurium in HEp-2 cell cultures pretreated with UV inactivated coxsackie virus. AB - The invasiveness of Salmonella typhimurium was significantly enhanced in cell cultures pretreated with UV-inactivated virus. During the first 3 hours of virus infection there was no difference between the enhancement achieved with non inactivated and that achieved with UV-inactivated virus. After 4 and 5 hours pretreatment the effect of non-inactivated virus was more pronounced than that of UV-inactivated virus. The results indicate that during the early period of virus infection the enhancement of bacterial invasiveness by pretreatment with virus is the result of a direct interaction between the virus and the cell membrane. During the later phase of viral reproduction, viral RNA induced alteration of the cell metabolism, and these altered products might be involved in the interaction. PMID- 2984879 TI - Effects of lithium and antidepressant treatment on neurotransmitter receptors. International Symposium on the Neuropharmacology of Manic-Depressive Illness. Copenhagen, June 20-22, 1983. PMID- 2984880 TI - Effects of lithium and antidepressant treatment on neurotransmitter receptors. Introduction. PMID- 2984881 TI - Regulation of serotonin neurotransmission by antidepressant drugs and electroconvulsive shock: the fall and rise of serotonin receptors. PMID- 2984882 TI - Serotonin-selective antidepressants. PMID- 2984883 TI - A comparison of lithium effects on human brain and rat brain noradrenaline sensitive adenylate cyclase. AB - Lithium (Li) is a drug with numerous biochemical effects. Many of these biochemical effects occur only at high Li concentrations, or disappear after chronic Li treatment. Such effects are probably not related to Li's mechanism of therapeutic action. Inhibition of noradrenaline (NE)-sensitive adenylate cyclase has been proposed as a possible therapeutic mechanism of action for Li. Tolerance does not develop to this effect, which has been reported to occur reliably beginning at 2mM Li in rat cortex. Since 2mM Li is at the upper limit of therapeutic levels in humans, controversy has continued as to whether Li inhibition of NE-sensitive adenylate cyclase is a mechanism of Li's therapeutic action or a mechanism of Li toxicity. We hypothesized that human brain NE sensitive adenylate cyclase may be more sensitive to Li inhibition than rat brain NE-sensitive adenylate cyclase. Fresh cortical human grey matter was obtained from the edges of surgically removed brain tumors in seven patients. Results showed significant inhibition of NE-sensitive adenylate cyclase at 1mM Li. These results support the possibility that inhibition of NE-sensitive adenylate cyclase is a mechanism of Li's therapeutic action. PMID- 2984884 TI - Specific binding sites for cyclic antidepressants. PMID- 2984885 TI - The effect of antidepressants on circadian rhythms in brain neurotransmitter receptors. PMID- 2984886 TI - Opioid peptides and receptors in relation to affective illness. Effects of desipramine and lithium on opioid receptors in rat brain. AB - A brief review is given of clinical and experimental evidence supporting the notion that opioid peptides and opioid receptors play a role for the regulation of mood and activity, and that they could be involved in the pathophysiology of affective illness and the action of antimanic and antidepressant treatment modalities. We have carried out in vitro and in vivo studies on the effects of desipramine and lithium on opioid receptors in rat brain. In vitro desipramine inhibited the binding of 3H-enkephalinamide to neuronal membranes (P2-fractions) through mechanisms not yet known. Treatment with desipramine in vivo (10 mg/kg body weight/day) caused a down-regulation of 3H-enkephalinamide binding in the basal ganglia and the hippocampus, while no effects could be observed in the cerebral cortex and the rest of the forebrain. In vitro addition of lithium inhibited enkephalin binding to opioid receptors through a reduction in the number of binding sites, while the affinity remained unchanged or was changed only slightly. Treatment with lithium in vivo for three weeks with lithium doses providing serum lithium concentrations of about 1 mM also caused a down regulation in the number of opioid binding sites in the cerebral cortex, the hippocampus, and the basal ganglia, while no changes could be observed in affinity. The studies suggest that desipramine and lithium, both effective in the treatment of manic-depressive illness, may share certain actions on opioid receptors in the brain. PMID- 2984887 TI - Lithium, cyclic AMP and renal pathophysiology. PMID- 2984888 TI - Presynaptic and transynaptic mechanisms involved in the subsensitivity of rat cortical noradrenergic system after long-term antidepressant treatment. AB - Chronic treatment with antidepressants has been shown to produce a subsensitivity of noradrenergic neurons, both at presynaptic and postsynaptic sites. Important mechanisms, whereby the activity of noradrenergic neurons is regulated, could be the sensitivity of presynaptic alpha 2-adrenoceptors and the participation of transynaptic mechanisms involving other neurons. In this report we demonstrate that transynaptic factors involving the serotonergic system may be relevant to the regulation of the function of alpha 2-receptors in antidepressant chronically treated animals. In fact, we provide evidence of a markedly deminished responsiveness of noradrenergic neurons to an alpha 2-agonist (clonidine) or antagonist (mianserin) in biochemical and behavioral studies following serotonergic denervation with 5,7-dihydroxytryptamine. These results indicate that a functional interrelationship between serotonergic and noradrenergic systems might play an important role in the adaptive changes which bring the noradrenergic neurons to a lower level of activity after chronic antidepressant administration. PMID- 2984889 TI - Platelet alpha-adrenergic receptors in depression: ligand binding and aggregation studies. PMID- 2984890 TI - Alterations in monoamine-containing neuronal function due to administration of antidepressants repeatedly to rats. PMID- 2984891 TI - Effects of lithium and antidepressants on electrophysiological and biochemical processes in the CNS. PMID- 2984892 TI - Alpha-adrenergic receptor changes during antidepressant treatment. PMID- 2984893 TI - Beta adrenergic receptor function in depression and the effect of antidepressant drugs. AB - It has been suggested that alterations of monoamine receptor sensitivity in the central nervous system may be associated with some forms of affective illness. It has been observed by several investigators that chronic treatment with antidepressant drugs causes down regulation of NE receptor coupled adenylate cyclase and beta adrenergic receptor binding in rat brain. This observation has led to the suggestion that the therapeutic effects of antidepressant drugs may be related to the changes in the responsivity of beta adrenergic receptors. In order to examine if depressive illness may be associated with altered beta adrenergic function, we studied adenylate cyclase and its responsiveness to norepinephrine and isoproterenol in the leukocytes obtained from patients with psychiatric illness and normal controls as an index of beta adrenergic receptor function. We also studied the effects of antidepressant drugs, in vitro, on isoproterenol sensitive leukocyte adenylate cyclase. We observed that norepinephrine and isoproterenol sensitive leukocyte adenylate cyclase in depressed patients are significantly decreased as compared to normal controls. Our results appear to have been replicated by another group of investigators. We also observed that certain antidepressant drugs potentiate isoproterenol stimulated accumulation of cyclic AMP in human leukocytes. This potentiation was most pronounced in the case of iprindole. These results thus indicated a decreased beta adrenergic receptor function in patients with depressive illness. Whether or not such decreased receptor function is associated with depressive illness or is a manifestation of some other changes unrelated to the illness is not clear. Our results also indicate that some antidepressant drugs may enhance adrenergic transmission by potentiating the effects of neurotransmitters on beta adrenergic receptors. PMID- 2984894 TI - Influence of lithium in vitro and in vivo on the catecholamine-sensitive cerebral adenylate cyclase systems. PMID- 2984895 TI - Skeletal muscle and hormonal adaptation to physical training in the rat: role of the sympatho-adrenal system. AB - The main purpose of the present study was to test the hypothesis that adrenergic stimulation of muscle fibres during exercise is a major stimulus for the training induced enhancement of skeletal muscle respiratory capacity. Therefore, Sprague Dawley rats either underwent bilateral surgical ablation of the adrenal medulla or were sham-operated. Furthermore, unilateral surgical extirpation of the lumbar sympathetic chain was performed. Half of the rats were then trained for 12 weeks by swimming (up to 5.5 h X day-1, 4 days X week-1) and the remaining rats were sedentary controls. In the gastrocnemius muscle, training significantly increased the mitochondrial enzymes citrate synthase, succinate dehydrogenase, cytochrome c oxidase, and 3-hydroxyacyl-CoA dehydrogenase. In sham-operated rats, the increases were 40%, 43%, 66%, and 25%, respectively, in legs with intact sympathetic innervation. The training-induced enzyme adaptation after adrenodemedullation and/or sympathectomy was not significantly lower than these control values. In sham-operated rats, training decreased resting plasma insulin and glucagon levels and increased liver glycogen content. Similar changes were induced by adrenodemedullation, but training did not augment these changes in adrenodemedullated rats. In conclusion, the data suggest that neither adrenomedullary hormones nor local sympathetic nerves are prerequisites for the training-induced increase in muscle mitochondrial enzymes. The training-induced decline in resting plasma insulin and glucagon levels in intact rats may be mediated by adrenomedullary hormones. PMID- 2984896 TI - Actions of enprofylline in the rat hippocampus. AB - Enprofylline is a novel xanthine with a low adenosine antagonistic potency, but with a good bronchodilator activity. In order to further elucidate its central nervous actions we have studied its effects in the rat hippocampus, both electrophysiologically and biochemically. Adenosine was found to depress evoked field excitatory post-synaptic potentials (EPSPs) in the rat hippocampus. The effect of adenosine was antagonized by theophylline but not by enprofylline. Both xanthines caused a slight increase per se of the field EPSP amplitude. Cyclic AMP accumulation in the rat hippocampus was stimulated by the adenosine analogue NECA, by isoprenaline and by noradrenaline in the presence of propranolol. The cyclic AMP accumulation induced by these agents was not significantly altered by omitting calcium. Enprofylline antagonized the effect of NECA and noradrenaline (after beta-blockade) but not that of isoprenaline. The IC50 of enprofylline against NECA was about 20 mumol X 1(-1). The inhibitory effect of enprofylline against NECA induced cyclic AMP accumulation was not affected by omitting calcium or by the calcium channel antagonist felodipine, but was decreased by the calmodulin inhibitor stelazine and the intracellular calcium chelator Quin-2. The results show that enprofylline is much more potent as an inhibitor of adenosine A2-receptor-mediated cyclic AMP accumulation than against adenosine A1-receptor mediated inhibition of field EPSP. The difference may depend on a role of a special pool of calcium in the former response. PMID- 2984897 TI - Characterization of pre- and post-junctional adenosine receptors in guinea-pig ileum. AB - The receptors involved in adenosine-induced modulation of cholinergic neuroeffector transmission in guinea-pig ileum were explored by means of the non selective stable analogue, 2-chloroadenosine and analogues with preference for AI receptors, L-N6-phenylisopropyladenosine (L-PIA), and A2 receptors, 5'-N ethylcarboxamideadenosine (NECA) and D-N6-phenylisopropyladenosine (D-PIA). 2 chloroadenosine, L-PIA and NECA were equipotent in inhibiting contractile responses to nerve stimulation, whereas D-PIA exerted a similar activity only in high concentrations. The release of acetylcholine induced by nerve stimulation was inhibited to a similar degree by NECA, L-PIA and D-PIA. The phosphodiesterase inhibitor, ZK 62.7II, and the activator of adenylate cyclase, forskolin, enhanced the inhibitory effect of NECA, but not that of L-PIA, on contractile responses to nerve stimulation. Only NECA inhibited contractions induced by direct muscle stimulation and ZK 62.7II enhanced this inhibition. It is concluded that adenosine inhibits the neuroeffector transmission in guinea-pig ileum mainly by a prejunctional, cAMP-independent, mechanism, involving AI receptors and a supplementary activation of post-junctional A2 receptors. In addition there may be a prejunctional inhibitory effect of high agonist concentrations, exerted via A2 receptors and influenced by the prevailing levels of cAMP. PMID- 2984898 TI - Prognostic factors of papillary, follicular and medullary carcinomas of the thyroid gland. Retrospective multivariate analysis of 216 patients with a median follow-up of 11 years. AB - Various prognostic factors have been tested in multivariate analyses of 216 patients with papillary, follicular or medullary thyroid carcinomas without initial distant metastases. The median follow-up time was 11 years. The patient's sex was not found to be a significant predictor. Age at diagnosis seemed to be an important predictor for papillary as well as for follicular carcinomas, but when deaths in intercurrent disease were estimated, marked cellular atypia and tumour invasion beyond the thyroid capsule proved to be more important predictors. For medullary carcinomas tumour invasion beyond the thyroid capsule was the only significant predictor. PMID- 2984899 TI - Recombinant leukocyte A interferon as single agent therapy or in combination with cimetidine in patients with advanced colo-rectal carcinoma. A phase II investigation. AB - Recombinant leukocyte A interferon used as single agent therapy (7.5 X 10(6) units/m2 three times weekly intramuscularly) showed very limited clinical efficacy in patients with advanced colo-rectal carcinoma. No objective tumour regressions were documented in 15 evaluable patients, although three patients demonstrated a stable disease status after 12 weeks of treatment. Neither were any objective tumour regressions registered in 13 patients during subsequent combined therapy with Recombinant Leukocyte A Interferon and cimetidine (1 000 mg/day orally). Two of the 3 patients maintained their stable disease status. No change in natural killer (NK) cell activity or in antibody dependent cellular cytotoxicity (ADCC) of peripheral blood mononuclear cells was seen during the two treatment periods. PMID- 2984900 TI - Testicular carcinoma. A curable malignancy. AB - The progress in the management of testicular germ cell tumours is reviewed. A marked improvement of the treatment results has been obtained in non-seminomas, especially by the use of cis-platinum based chemotherapy. At present long term survival can be expected in 85 per cent of all non-seminomas and in 95 per cent of all seminomas after adequate treatment. The natural history of the disease, symptomatology, diagnostic procedures, staging and different treatment modalities, as surgery, radiation therapy, chemotherapy and their combination, are discussed. PMID- 2984901 TI - Invasive squamous cell carcinoma of the uterine cervix. VIII. Survival and malignancy grading in patients treated by irradiation in Lund 1969-1970. AB - The correlation between certain prognostic factors and the 10-year survival/lethality rate was analysed in 168 patients with invasive squamous cell carcinoma of the uterine cervix stage I A-II B treated by radiation therapy from 1969 through 1970. These factors included a malignancy grading system (MGS) consisting of 8 items: structure (P1), differentiation into cell type (P2), nuclear polymorphism (P3), mitosis (P4), mode of invasion (P5), stage of invasion (P6), vascular invasion (P7), and host-cellular response (P8). Histologic differentiation (1) and differentiation into cell type (11), the patient's age, the year of admission, the clinical stage (FIGO), and irradiation were also analysed. Many of these factors were correlated to the prognosis. However, the MGS system was superior as a predictive factor. Patients with a low MGS score had an extremely good survival rate at both the 5- and 10-year controls. The patients with a high MGS score had approximately 55 per cent lethality at 10 years. The MGS was significantly superior to each separate item as well as to each predictive factor (p less than 0.05). Further, no other important predictive factor could be identified after the MGS had been included in the multivariate analyses. PMID- 2984902 TI - Radiation treatment of primary carcinoma of the vagina. Patterns of failures after definitive therapy. AB - Patterns of failures after definitive radiation therapy were analyzed in 88 patients with primary carcinoma of the vagina treated between 1957 and 1975. The majority of the local failures in early stages of the disease (I, II and a few III) were due to inadequate treatment either by external beam therapy or brachytherapy. In some cases the inadequate treatments were unavoidable (previous radiation treatment) and in a few they were due to poor brachytherapy technique. Distant failures are still a problem and need to be given further attention. PMID- 2984903 TI - Relation between tumour size and plasma prolactin levels in premenopausal patients with breast carcinoma. A preliminary report. AB - In thirty-one premenopausal patients with carcinoma of the breast the plasma prolactin was measured after mastectomy. A highly significant correlation between tumour size and plasma prolactin levels (p less than 0.002) was observed after adjustment for age at diagnosis and parity. At the time of the prolactin determination no clinical signs of metastatic disease were evident, suggesting that the prolactin levels were unrelated to the tumour burden. PMID- 2984904 TI - Lethal acute gamma radiation accident at Kjeller, Norway. Report of a case. AB - On September 2, 1982, a 64-year-old man accidently received a heavy dose of gamma radiation in a plant for sterilization by radiation. He developed an acute radiation syndrome. The leukocyte count rapidly diminished to low values, and extensive chromosome injuries could be demonstrated in cultured lymphocytes. He was hospitalized in an isolated room, and received large doses of antibiotics, and transfusions of leukocytes, platelets and blood. He died anuric on the 13th day. From spectroscopic analyses of electron-spin resonance in irradiated material the mean dose in the whole body was estimated to be 22.5 +/- 2 Gy. The course of the illness and the findings at autopsy are briefly described. PMID- 2984905 TI - Selective shielding of a p-Si detector for quality independence. AB - p-silicon semiconductor detectors were partially shielded with lead filters of various geometries with the purpose to minimize the quality dependence in assessments of depth dose distributions in large fields of cobalt radiation. It is shown that a lead shielding of the detector, with the front side excluded, reduced the signal error from about 3 to less than one per cent of the maximum signal as compared with an ionization chamber. Shielding was also done by encapsulating the detector in a mixture of wolfram powder and epoxy resin, with similar results. The signal received at the surface was not affected by this filter geometry. The directional dependence was investigated and was far from uniform for the wolfram-shielded detector. This will however not cause any practical problems when the detector is oriented towards the radiation source. PMID- 2984906 TI - Characteristics of a p-Si detector in high energy electron fields. AB - Comparison of depth ionization distributions from a silicon semiconductor detector and depth dose curves from a plane parallel ionization chamber show that a semiconductor detector of p-type is well suited for relative electron dosimetry in the energy range of 6 to 20 MeV in Ep,0. Maximum deviations of the order of 1.5 per cent and of 1 mm were obtained down to a phantom depth of about 1 mm. The directional dependence of the detector was about 4 per cent. PMID- 2984907 TI - Electron beam dose planning using Gaussian beams. Energy and spatial scaling with inhomogeneities. AB - The penetration of an electron beam in matter is characterized by only two material dependent parameters within the range of validity of the small angle diffusion equation. These parameters characterize the relative importance of multiple scattering and energy loss for a given material and energy. By comparison with more accurate Monte Carlo calculations it is shown that the same two parameters can be used to scale the dose distribution from one material and energy to an equivalent energy in an arbitrary material with high precision also outside the limits of the small angle approximation. This scaling procedure has been generalized to the non-uniform case with plane parallel slab inhomogeneities by assuming the angular distribution on each side of the interfaces to be equal. The generalized recursive scaling procedure is finally tested by measurements on almost point monodirectional beams in water with inserted aluminum plates of varying thicknesses. The agreement with the scaling procedure is of the order of one per cent with regard to the radial dose profiles. PMID- 2984908 TI - Irradiation induced cell death as related to cell cycle. AB - Non-viable cells can be identified by their low density. In a Percoll density gradient we found in ascites tumour cells three fractions of cells. The first of these fractions at a low density of 1.020 g/ml contained morphologically dead cells and cells with a plating efficiency of less than 10 per cent, while the other two fractions contained pure viable cells. The proportion of cells in the cell cycle of the three fractions was measured using DNA flow cytometry. After whole-body irradiation with 5 Gy roentgen rays of ascites-bearing mice we calculated the dead cells in the various parts of the cell cycle up to 72 hours. The proportion of non-viable cells increased from about 5 per cent to about 20 per cent 20 to 25 hours after irradiation. The majority of these cells was in G1 and released from the irradiation induced G2-blockage. In addition, an increased number of non-viable S-phase cells preceded this G1 cell by some hours. These cells may represent cells damaged in G1 and S-phase at the time of irradiation and incapable of progressing further in the cell cycle. PMID- 2984910 TI - Surgical approach to large thalamic gliomas. AB - Fifteen cases of large thalamic gliomas were treated by laser surgery and CUSA under X8 magnification and subsequently they were subjected to radiotherapy. More than 50% of the tumours were located in the dominant hemisphere and therefore an interhemispheric approach was carried out to avoid disturbances of speech and field defects. Nine patients had astrocytomas grade II and the remaining six had astrocytomas grades III-IV. The quality and survival has been gratifying in the less malignant gliomas. The operative technique and use of Laser and CUSA is highlighted. PMID- 2984909 TI - Investigations of 90Sr in dogs. I. Pathogenesis of radiation-induced bone tumors. AB - Purebred beagle dogs given 90Sr and unirradiated controls were studied for over two decades. Pregnant females were fed different doses of 90Sr from day 21 post conception until the offsprings reached an age of 540 days. In an additional experiment two dose levels were given in a single intravenous dose to dogs 540 days old. Radiographically the earliest skeleton lesions were characterised by small linear, solitary, cortical lucencies. These as well as tumors were more frequently noted in the higher exposure levels. They affected the appendicular skeleton almost as frequently as the axial skeleton. The lesions were predominantly found in the diaphysis, at the angle and near the acetabulum in the tubular bones, mandible and pelvis, respectively. The lesions within the diaphysis originated in the cortical bone. Histologically these lesions were characterised by different types of porosities. These could be empty or filled by a defect and/or immature, dysplastic fibrous repair tissue, within the frame of which malignant transformations seemed to take place as evidenced by malignant clones and micro-osteosarcomas. A comparison is made of the histologic events in dogs and mice and a tentative pathogenesis of 90Sr induced bone tumors is discussed. PMID- 2984911 TI - Cerebral magnetic resonance image synthesis. AB - The authors previously described magnetic resonance (MR) image synthesis, a process that enables the investigator to manipulate imaging parameters retrospectively and generate or "synthesize" the image that corresponds to various arbitrary scanning factors. They demonstrate the validity and utility of synthetic spin-echo images in cerebral imaging. As a test of their method, spin echo images are synthesized for echo times identical to those of the original acquired images as well as for alternate values. Subjectively, the quality of synthetic and acquired images is comparable. It is shown quantitatively for several tissue types that the reconstructed synthetic signal matches the acquired signal within the uncertainty of the acquired images. Observed and measured noise levels in the acquired and synthetic images are comparable. Because of a signal averaging effect, the synthetic images can have a higher signal-to-noise ratio than the source images, thereby providing improved boundary definition. Applications of MR image synthesis are discussed with respect to potential reduction in scanning time. The advantages of image synthesis versus analysis of computed images are discussed. PMID- 2984913 TI - A parallel study of enalapril and captopril and 1 year of experience with enalapril treatment in moderate-to-severe essential hypertension. AB - Angiotensin converting enzyme (ACE) inhibitors, enalapril (5 to 20 mg twice daily) or captopril (25 to 100 mg thrice daily) and matching ACE inhibitor placebos were given to 32 moderate-to-severe essential hypertension patients who were already on 50 mg hydrochlorothiazide daily. Alpha-methyldopa (250 to 500 mg twice daily) was given to 16 patients following 6 weeks of ACE inhibitor therapy. Both enalapril and captopril significantly (p less than 0.05) lowered the supine and upright blood pressures (BPs) (acutely and long-term) without significant reflex heart rate changes. The BPs of enalapril patients were, however, significantly lower (supine diastolic p less than 0.03, supine systolic p less than 0.05, and upright diastolic p less than 0.04) than those of captopril patients when compared by repeated measures of analysis of variance. Eleven enalapril patients have been followed for 1 year with continued BP control. Skin rash occurred in one captopril patient and reversible renal insufficiency developed in two enalapril patients during the first 16 weeks. It is concluded that (although both ACE inhibitors lowered BP, enalapril was more effective than captopril and twice-daily enalapril was well tolerated during 52 weeks of treatment. PMID- 2984912 TI - House calls: current status and rationale. AB - House calls play a limited but important role in modern family practice. Family physicians make an average of two to six home visits a week for various reasons, including acute illness, terminal care, death pronouncement and hospital follow up. The rationale for house calls includes enhancing the doctor/patient relationship, providing medical therapy, evaluating and reinforcing the patient's support system, and making an ecologic or family assessment. PMID- 2984915 TI - The relations between vitamin D2 and D3 in the diet and plasma 25OHD2 and 25OHD3 in elderly women in Great Britain. AB - Vitamin D2 and D3 intake and plasma 25OHD2 and 25OHD3 were measured in 70 elderly women; 13 living at home and 57 long-stay patients with no access to sunlight. Vitamin D2 intake and plasma 25OHD2 were correlated in the whole group (p less than .005) and vitamin D3 intake and plasma 25OHD3 and total D intake and total 25OHD were significantly correlated (p less than .005) in the patients. In the whole group the plasma 25OHD2 increased by 4.5 nmol/l for every 1 microgram increase in vitamin D2 intake. This was also the increase observed in a longitudinal study of vitamin D2 supplements in 11 patients. Vitamin D intake is a significant determinant of plasma 25OHD and the relation between them suggests that stores of vitamin D can be maintained at 20 nmol/l in the elderly by a daily intake of 4 micrograms of vitamin D, even in the absence of sunlight. PMID- 2984914 TI - Cardiovascular effects and regional blood flow distribution associated with angiotensin converting enzyme inhibition (captopril) in essential hypertension. AB - Systemic and regional hemodynamics and cardiac structural changes were studied in 12 patients with mild to moderately severe essential hypertension before and then 90 minutes and 12 weeks after administration of captopril. Mean arterial pressure was reduced from 111 mm Hg to 96 mm Hg (p less than 0.001), and this was mediated through a fall in total peripheral resistance from 26 +/- 2 units to 23 +/- 2 units (p less than 0.01). The decreased total peripheral resistance was distributed to all circulations studied: kidney, skeletal muscle, skin and the splanchnic organs. Furthermore, left ventricular (LV) mass index diminished without altering myocardial contractility at rest. Thus, captopril lowered arterial pressure through systemic arteriolar dilatation in patients with mild to moderately severe essential hypertension and also reduced LV mass even in patients without evidence of LV hypertrophy. PMID- 2984916 TI - Estimation of dietary fiber supply. PMID- 2984917 TI - Histocytologic diagnosis of pancreatic cancer by percutaneous aspiration biopsy under ultrasonic guidance. AB - Percutaneous aspiration biopsy of the pancreas using a heparinized 22-gauge fine needle was performed under ultrasonic guidance in five patients with benign pancreatic diseases and in 18 patients with pancreatic cancer. Using a heparinized needle and syringe, it was possible to make good smears containing abundant tumor cells and to obtain small tissue specimens. Using egg albumin as binding material, a new cell-block technic was developed to conveniently obtain histologic specimens. In this way, a correct diagnosis was made cytologically in all 23 patients suspected of having a pancreatic malignancy. Histologic specimens were obtained in 22 (95.6%) our of 23 patients. A correct diagnosis was established histologically in all patients from whom histologic materials were obtained. This procedure thus has proved a very reliable method for diagnosing pancreatic cancer. PMID- 2984919 TI - Use of sonication for viral isolation. AB - A viral passage method using sonication to obtain cell-free virus was compared with the conventional viral cell culture passage technic. The recovery of 121 varicella zoster virus and cytomegalovirus isolates in human fibroblast vials from sonicated versus nonsonicated passage suspensions was studied. These fibroblast vials with cytopathic effect were identified using monoclonal fluorescent antisera. Twenty-eight (29%) of cytomegalovirus isolates were recovered only from sonicated passages, and an additional 28 isolates had faster recovery and/or a greater number of viral plaques from sonicated passages. Sonication did not increase significantly the varicella zoster virus recovery rate. PMID- 2984918 TI - Antibodies to human T-cell lymphotropic virus-I membrane antigens and inverted T4/T8 ratios in hemophiliacs. AB - Antibodies to the membrane antigens of human T-cell lymphotropic virus-I (anti HTLV-MA) have been detected in patients with the acquired immune deficiency syndrome (AIDS) and in patients with hemophilia. The authors examined sera from 71 AIDS patients and 46 hemophiliac children for the presence of anti-HTLV-MA using an indirect membrane immunofluorescence assay with flow cytometry analysis. Thirty-seven of the 71 (52%) AIDS patients and 7 of the 46 (15%) hemophiliac patients had high titered anti-HTLV-MA, using a T-lymphoid cell line infected with the leukemia virus. None of the 78 control subjects had high titered antibody. All seven hemophiliac patients with elevated anti-HTLV-MA used Factor VIII concentrates, and all had inverted T-lymphocyte helper-suppressor (T4 [Leu 3]/T8 [Leu 2]) ratios. No correlations were found between inverted T4/T8 ratios and antibody to cytomegalovirus, Toxoplasma gondii, or hepatitis B. This work supports contentions that HTLV-like organisms cause AIDS and that these organisms are transmitted by blood products such as Factor VIII concentrate. PMID- 2984920 TI - Babesiosis: problems in diagnosis using autoanalyzers. AB - A 76-year-old white man previous diagnosed as having Waldenstrom's macroglobulinemia continued with persistent fevers and sweats for two and a half years. Recently, repeated automated differentials during 11 days of hospitalization failed to note any intracellular inclusions in the RBCs. Blood sent to the Microbiology Laboratory was noted to contain Babesia species. A review of the hematology slides revealed that Babesia species was present on all the slides the analyzer had screened. This failure to note infected RBCs may pose serious diagnostic problems. PMID- 2984921 TI - Pseudotumor resulting from atypical mycobacterial infection: a "histoid" variety of Mycobacterium avium-intracellulare complex infection. AB - A 54-year-old immunosuppressed cardiac transplant recipient with a six-month history of progressive swelling of the hand, with nodules and linear lymph node chain enlargement, diagnosed as a sporotrichoid Mycobacterium avium intracellulare pseudotumor is described. The microscopic features closely resembled the previously described histoid variety of lepromatous leprosy. Routine hematoxylin and eosin staining suggested a spindle cell neoplasm rather than an infectious or inflammatory process. An infectious etiology was pursued on the basis of the clinical setting. PMID- 2984922 TI - Peculiar cytoplasmic inclusions in acute lymphoblastic leukemia: cytochemical and electron microscopic observations. AB - In acute lymphoblastic leukemia (ALL) of a 19-year-old woman, blast cells in the blood and bone marrow contained peculiar large cytoplasmic inclusions. The inclusions, measuring 0.5-3.5 micron in diameter, were stained with methylene blue, toluidin blue, pyronin, and periodic acid-Schiff. Activities for acid phosphatase and alpha naphthyl butyrate esterase were not detected in the inclusions. The cytochemical findings indicated that the inclusions contained basophilic material and polysaccharides and that they were not lysosomal in nature. In electron microscopy, the inclusions were membrane bound and contained densely packed microvesicles, measuring 40-70 nm in diameter; amorphous electron dense material; and whorled membranous figures. The microvesicles often had a nucleoid-like core and resembled viruses in size and structure. PMID- 2984923 TI - Laxative use in constipation. American College of Gastroenterology's Committee on FDA-Related Matters. PMID- 2984924 TI - Localization of DNA sequences in region Xp21 of the human X chromosome: search for molecular markers close to the Duchenne muscular dystrophy locus. AB - Panels of somatic cell hybrid lines carrying various structural rearrangements of the human X chromosome short arm were analyzed with 21 X-chromosome-specific cloned DNA fragments. We mapped these molecular markers to five different regions of the short arm of the X chromosome. The results were confirmed by gene-dosage studies of human lymphoblasts with structurally abnormal X chromosomes. The ornithine transcarbamylase gene and four anonymous DNA sequences map within band Xp21, flanking the presumed locus for Duchenne muscular dystrophy. PMID- 2984925 TI - DNA restriction fragment analysis of the proopiomelanocortin gene in schizophrenia and bipolar disorders. AB - The method of DNA restriction fragment analysis using gene probes for the proopiomelanocortin (POMC) gene was employed to detect possible molecular variation in the POMC gene in schizophrenia and bipolar illness. No gross structural abnormalities in restriction fragments were observed with the set of restriction enzymes used. Two allelic restriction sites were observed giving rise to fragment length polymorphisms. One of these is a new polymorphism, not previously reported, which will be of value as a linkage marker. The associations between the two DNA polymorphisms that are closely linked to the POMC gene and both schizophrenia and bipolar disorder were investigated. No association was found, thus adding weight to the evidence that there are no alterations in the POMC gene in schizophrenia and bipolar illness. PMID- 2984926 TI - A molecular basis for discrete size variation in human ribosomal DNA. AB - The tandemly repeated human ribosomal RNA (rRNA) genes contain a region of size heterogeneity that is present in the nontranscribed spacer of every individual examined. This heterogeneity has been previously examined by Southern analysis of BamHI-digested human DNA. Using a ribosomal DNA (rDNA) probe specific for the 3' end of the 28S rRNA gene, at least four discrete sizes of BamHI fragments were seen in human populations. Molecular analysis of the cloned DNA from this region reveals tandem duplication of a segment of spacer rDNA located 388 base pairs (bp) 3' to the end of the 28S ribosomal RNA gene. Five hundred fifty bp of DNA, flanked on either side by a 150-bp repeated element, is either duplicated or deleted to produce a series of spacers that differ in size by 850 bp. These duplications/deletions appear to be the product of unequal homologous exchange, mediated by the small repeated element. Thus, human rDNA fragments cloned in lambda vectors and propagated in E. coli generate the same apparent size variation seen in genomic DNA. This study suggests that unequal homologous exchange is the molecular basis for the observed length heterogeneity in the spacer rDNA and may be a common mechanism for the generation of human genetic diversity. PMID- 2984927 TI - Mapping of X-linked Becker muscular dystrophy through crossovers identified by DNA polymorphisms and by haplotype characterization in somatic cell hybrids. AB - The analysis of 10 X-linked DNA polymorphisms (five mapping on the short arm and five on the long arm) in two Becker muscular dystrophy pedigrees has been used to localize this gene in the known sequence of DNA polymorphic markers on the X chromosome. In the first pedigree, the carrier mother, whose phase for Becker and for five informative polymorphisms is known, has transmitted a double recombinant X chromosome to one of her two affected sons. The discordance between these two affected brothers for four of the five informative polymorphisms indicates that the Becker gene is located between RC8 or D2 on one side and pDP34 on the other. In the second pedigree, where the maternal grandfather is dead and two maternal first cousins are affected, the phase of DNA polymorphic alleles has been identified in somatic cell hybrids resulting from the fusion of hamster fibroblasts with lymphocytes of the mothers and aunt of the patients. The discordance between the two first cousins for two of the four informative DNA polymorphisms is best explained by the occurrence of a single recombination in the X chromosome carried by one of them. This result further restricts the localization of the Becker gene to a region of the short arm delimited by B24 and L 1.28. Regional and fine gene mapping through the approach described in this paper should become useful in the future for X-linked as well as for autosomal genes. PMID- 2984928 TI - Human salivary proline-rich protein genes on chromosome 12. AB - A DNA probe (PRP1) for the proline-rich protein (PRP) genes was used to analyze the segregation of human PRP genes in human X mouse somatic cell hybrids. Endonuclease restriction analysis of 22 independent hybrid clones segregating human chromosomes demonstrated that PRP genes segregate with human chromosome 12 only and were therefore assigned to that chromosome. The PRP1 probe should prove useful for further mapping studies of human chromosome 12. PMID- 2984929 TI - Competitive alternatives to hospital inpatient care. AB - Reasons for the heightened national interest in alternative health-care delivery systems, competitive forces and hospital-use patterns, and three major alternatives to inpatient hospital care are discussed. The Reagan Administration's policies on health-care incentives reform, the federal emphasis on prospective pricing systems, increased rate-regulation roles for the states, and growing employer concern over the cost of health benefits have all contributed to the increased interest in alternative delivery systems. Demand for inpatient hospital care is declining and will continue to decline. In addition to changes in the financing mechanisms for third-party payers, there are three broad competitive forces reducing the demand for hospital inpatient care. These are the growth and diversification of ambulatory care, the development of aftercare for chronically ill and elderly patients, and the expansion of alternative delivery systems. Ambulatory-care centers, preferred provider organizations, and health maintenance organizations are discussed, and their impact on hospitals, pharmacists, and the health-care system is evaluated. Trends in the development of these alternatives are described. Competition is forcing hospitals and health care providers to consider options such as aftercare, alternative delivery systems, ambulatory care, and multihospital systems in order to survive in today's health-care system. PMID- 2984930 TI - Second primary bronchogenic carcinomas after small cell carcinoma. PMID- 2984931 TI - Cigarette smoking in hypertensive patients. Blood pressure and endocrine responses. AB - The blood pressure and endocrine responses to cigarette smoking were studied in 19 hypertensive patients to determine whether smoking activates the renin aldosterone axis. Blood pressure rose from 140 +/- 7/99 +/- 3 (mean +/- SEM) to 151 +/- 5/108 +/- 2 mm Hg (p less than 0.01) within 10 minutes after smoking, and pulse rate also increased significantly (69 +/- 2 to 96 +/- 4 beats per minute). Plasma renin activity did not change but rose 15 minutes after ambulation. In contrast, plasma aldosterone and plasma cortisol levels increased significantly after smoking and peaked at 20 minutes: 13.9 +/- 0.9 to 20.2 +/- 2.0 ng/dl (p less than 0.01) and 10.2 +/- 1.0 to 22.0 +/- 2.2 micrograms/dl (p less than 0.01), respectively. These responses were closely correlated (r = 0.6467, p less than 0.01), suggesting a pituitary-adrenal mechanism is activated during smoking. Plasma ACTH levels rose from 58 +/- 6 to 87 +/- 10 pg/ml in 10 minutes (p less than 0.001) and to 90 +/- 14 pg/ml at 20 minutes (p less than 0.01). Total plasma catecholamine levels also rose from 468 +/- 60 to 624 +/- 73 pg/ml 10 minutes after smoking (p less than 0.01) and to 724 +/- 69 pg/ml (p less than 0.01) 15 minutes after ambulation. In hypertensive smokers, cigarette smoking is associated with an increase in blood pressure, pulse rate, and plasma ACTH, cortisol, aldosterone, and plasma catecholamine levels. The long-term significance of these acute hormonal changes in regard to blood pressure homeostasis and vascular disease in cigarette smokers remains to be determined. Smoking should be avoided prior to blood pressure and endocrine determinations. PMID- 2984932 TI - Anti-lymphocyte antibodies in patients with the acquired immune deficiency syndrome. AB - Human lymphotropic retroviruses lymphadenopathy-associated virus/human T lymphoma virus III have been recently implicated in the pathogenesis of the acquired immune deficiency syndrome (AIDS). The mechanisms leading to the complex immune deregulations of this disease, however, are still largely unknown. To investigate the possible presence of anti-lymphocyte antibodies, lymphocytes from a normal donor were incubated with serum samples from patients with AIDS. Substantial increases of up to 75 percent in the number of surface immunoglobulin-positive lymphocytes resulted from incubation with serum of patients with AIDS and AIDS related complex but not with serum of patients with non-AIDS-related diseases or of normal control subjects. Monoclonal antibodies to OKT11, OKT4, and OKT8 in conjunction with a double-labeling technique were then used to identify the type of surface immunoglobulin-positive lymphocytes. These experiments showed that binding of immunoglobulins to lymphocytes did not occur at random but was directed against OKT4- or OKT11-positive cells whereas OKT8-positive cells showed no detectable reactivity. The results of these studies indicate that patients with AIDS and AIDS-related complex have circulating antibodies capable of reacting selectively with a population of T cells that is predominantly composed of helper cells and does not include suppressor cells. The augmentation of surface immunoglobulin-positive lymphocytes in the patients studied consistently paralleled the marked decreases of the helper/suppressor cell ratios and the presence of circulating anti-human T lymphoma virus III antibodies. Binding of antibodies to the surface of helper T cells may be a determining event in the pathogenesis of this disease. PMID- 2984933 TI - Development of hypercalcemic hyperparathyroidism after long-term phosphate supplementation in hypophosphatemic osteomalacia. Report of two cases. AB - Orally administered phosphate supplements are the mainstay of therapy for hypophosphatemic osteomalacia of diverse causes and are generally believed to be free from harmful side effects. Two cases are reported, however, in which long term therapy (14 and 10 years, respectively) resulted in hypercalcemic hyperparathyroidism associated with surgically proved adenomatous hyperplasia. This complication occurred despite concomitant treatment with pharmacologic doses of vitamin D. Thus, long-term oral phosphate therapy can produce tertiary hyperparathyroidism in susceptible patients. PMID- 2984934 TI - Noncaseating pulmonary granulomas associated with small cell carcinoma of the lung. AB - Noncaseating pulmonary granulomas are rarely associated with primary carcinoma of the lung. The patient described herein presented with constitutional symptoms and nodular pulmonary infiltrates associated with noncaseating granulomas without evident neoplasm in both transbronchial and open lung biopsy specimens. Despite corticosteroid therapy for presumed sarcoidosis, chest roentgenographic findings worsened and repeated transbronchial biopsy 12 months after the onset of initial symptoms revealed small cell carcinoma of the lung. Twenty-two months after initiation of chemotherapy, the patient is well with no evidence for carcinoma. PMID- 2984935 TI - Immunotherapeutic studies of different clonal lines of herpesvirus-induced fibrosarcoma in a syngeneic rat. AB - Transformation of White Buffalo rat embryonic cells by dye-light-inactivated herpesvirus type 2 resulted in the development of two clones of transformed cells (G2 and rat fibrosarcoma) with significantly different tumorigenic capabilities. The G2 cell line was initially nontumorigenic, while the rat fibrosarcoma line was very highly tumorigenic in rats. These significant differences in transformed cell clones from the same initial culture offered an opportunity to study aspects of the immunobiology of oncogenicity. The development of fibrosarcomas in immunoincompetent nude mice with the same early passages of the G2 cell line which were nontumorigenic in the immunocompetent rat suggested that immunologic resistance developed more effectively in the competent host against the G2 line than against rat fibrosarcoma cells. Syngeneic rats which were first exposed to the early passage nontumorigenic G2 cells were completely protected against tumor development by the rat fibrosarcoma cell lines. In subsequent in vitro passages of the G2 cell line, it lost its ability to protect against rat fibrosarcoma challenge and gradually became oncogenic in rats. Modification of antigenic exposure, accomplished by treating the G2 cells with cholesteryl hemisuccinate, resulted in an increase in the protection by these cells and a delay in tumor development. PMID- 2984936 TI - Uterine sarcoma: analysis of prognostic variables in 71 cases. AB - The histories of 94 patients with a diagnosis of uterine sarcoma, treated from 1962 to 1982, at the Medical College of Virginia Hospital were reviewed. Histologic features were studied by one of the authors (V.S.), and cases that did not meet strict pathologic criteria were rejected. The 71 patients with uterine sarcoma accepted for this study had a survival rate of 22.5% from 1 to 11 years. Survival rates were similar for leiomyosarcoma, mixed mesodermal tumor, and endometrial stromal sarcoma. Clinical staging had some predictive value, since 45 patients with Stage I disease had a 27% survival rate, and 26 patients with Stages II, III, and IV disease had only a 12% survival rate (p less than 0.05). The type of treatment had no demonstrable effect on final outcome but did influence the recurrence patterns. Chemotherapy (used in 25 patients) was neither of benefit when used as adjuvant therapy nor effective in prolonging survival in patients with recurrence (mean survival, 5.4 months). PMID- 2984937 TI - Corticotropin-releasing factor--induced adrenocorticotropic hormone release in the sheep fetus: blockade by cortisol. AB - We administered intravenous injections of synthetic ovine corticotropin-releasing factor to chronically cannulated sheep fetuses and monitored fetal plasma adrenocorticotropic hormone and cortisol concentrations. The three doses of corticotropin-releasing factor used (10, 100, or 1000 ng X kg-1) increased fetal plasma adrenocorticotropic hormone; fetal plasma cortisol levels rose with the highest dose of corticotropin-releasing factor. Administration of corticotropin releasing factor at these concentrations did not change fetal heart rate or blood pressure. Elevation of fetal plasma cortisol levels to 40 to 80 ng X ml-1 by infusions of the steroid blocked the adrenocorticotropic hormone responses to all three doses of corticotropin-releasing factor. These data indicate that corticotropin-releasing factor can increase plasma adrenocorticotropic hormone concentrations in the late-gestation fetus and that these increases can be blocked by elevations in fetal plasma cortisol levels within a physiologic range. This suggests that cortisol modulates adrenocorticotropic hormone release by the fetal pituitary gland late in gestation. PMID- 2984938 TI - Compressive optic neuropathy induced by intranasal balloon catheter. PMID- 2984939 TI - Modulation of multiple neutrophil functions by preparative methods or trace concentrations of bacterial lipopolysaccharide. AB - Human neutrophils were isolated from peripheral blood by four methods: 1) Ficoll Hypaque gradients and erythrocyte lysis, 2) plasma-Percoll gradients, 3) a "lipopolysaccharide (LPS)-free" method yielding 85% neutrophils, and 4) by centrifugation of cells prepared by Method 3 through a plasma-Percoll gradient to produce pure neutrophils. The use of the Ficoll-Hypaque method resulted in spontaneous change of cell shape, enhanced formyl-methionyl-leucyl-phenylalanine (FMLP)-stimulated release of superoxide anion, increased release of lysosomal enzymes upon subsequent FMLP stimulation, and reduced chemotactic responsiveness, by comparison with the other methods. These effects were not due to erythrocyte lysis by NH4C1 but were reproduced by exposure of neutrophils prepared by the "LPS-free" method or the use of plasma-Percoll gradients to 10-100 ng/ml LPS. Neutrophil change of shape and stimulated O-2 production were particularly sensitive markers of these effects. The effects of trace concentrations of LPS in the modulation of neutrophil function may have relevance to the pathophysiology of endotoxemia and its resultant tissue injury. PMID- 2984940 TI - In vitro induction of cytologic and functional differentiation of the immature human monocytelike cell line U-937 with phorbol myristate acetate. AB - The tumor-promoting ester 4 beta-phorbol 12-myristate 13-acetate (PMA) has been shown to induce the differentiation of the immature monocytelike cell line U-937 c in vitro into a heterogeneous population of cells, including small "dense" cells, large vacuolized or "foamy" cells, spindle-shaped cells, and cells with multiple filopodia ("stellate" cells). The effect of PMA was dose- and time dependent, the optimal conditions being 40-162 nM PMA for 48 hours. The minimum time of exposure to PMA to ensure further differentiation of U-937 cells was about 5 hours. The PMA-stimulated cells acquired morphologic, ultrastructural, and functional characteristics typical of cells of the monocyte/macrophage lineage. The PMA-treated U-937 cells became adherent, ceased to proliferate, and exhibited increased expression of monocyte-specific antigens (Leu-M2, - M3, HLADr), surface receptors (FcR, C3bR), enzymes (nonspecific esterase, transglutaminase), and ability to mediate chemotaxis, phagocytosis, superoxide anion production, and antibody-dependent cytotoxicity reactions. The induced cells lost their morphologic differentiation and ability to attach to surfaces and regained proliferative capacity upon repeated subculture in PMA-free media. PMID- 2984941 TI - Type C retrovirus production by pancreatic beta cells. Association with accelerated pathogenesis in C3H-db/db ("Diabetes") mice. AB - C3H.SW/SnJ females (haplotype H-2b) were mated with C3HeB/FeJ males (haplotype H 2k), which were heterozygous for the recessive mutation, diabetes (db). The object of the study was to analyze whether the H-2b haplotype conferred diabetes resistance to db/db males in the F2 generation. Severity of diabetes did not segregate with H-2 haplotype in this mouse model of diabetes. Instead, all F2 male mutants developed a much more severe diabetes syndrome than did "grandparental-type" C3HeB/FeJ-db/db males. Most surprisingly, unlike grandparental-type db/db females, which were uniformly resistant to the diabetogenic action of the db mutation, 75% of F2 db/db females developed severe diabetes. Ultrastructural comparison of beta cells in islets of these diabetic females versus those in the diabetes-resistant F2 mutants showed expression in the susceptible females of a type C retrovirus which budded extracellularly and intracellularly into vacuoles. No other islet endocrine cell type showed this expression, but intraislet exocrine cells as well as some ductal epithelial cells did produce type C particles. Macrophages were frequently observed in close association with the virus-expressing cell types. Failure of the diabetes acceleration factor(s) to segregate according to Mendelian expectations further suggested that type C retrovirus induction was linked to the accelerated pathogenesis of diabetes in the F2 generation. PMID- 2984942 TI - Immunoreactive ACTH and gamma-MSH in rat plasma during early stages of adrenal regeneration. AB - Recent evidence suggests that in addition to ACTH the pro-gamma-melanotropins play a role in controlling both the steroidogenic activity and growth of the adrenal cortex. By using the regenerating gland as a model for rapid adrenal growth, studies were carried out to monitor plasma ACTH and pro-gamma melanotropins during the acute phase following adrenal enucleation. Adrenal enucleated rats (experimental group) were uninephroadrenalectomized; controls were uninephrectomized or uninephroadrenalectomized. At intervals up to 96 h after surgery, animals from each of the three groups were killed under quiescent conditions at the low and high points of their circadian rhythm. The plasma concentrations of both ACTH and 11 K gamma-melanocyte-stimulating hormone (MSH) in the adrenal enucleated rats were markedly elevated as compared with either control group at each time point monitored. However, there were no significant differences in plasma ACTH or 11 K gamma-MSH between the two control groups, and the levels of plasma 6 K gamma-MSH remained unchanged in all three groups throughout the experiment. These data are consistent with a role for pro-gamma melanotropins in adrenal regeneration. PMID- 2984943 TI - Tauroursodeoxycholate prevents biliary protein excretion induced by other bile salts in the rat. AB - Biliary excretion of various proteins (5'-nucleotidase, alkaline phosphatase, lactate dehydrogenase, and albumin) was investigated in pentobarbital sodium anesthetized rats infused with different bile salts [taurocholate (TC), taurochenodeoxycholate (TCDC), and tauroursodeoxycholate (TUDC)]. A TCDC infusion at 0.4 mumol . min-1 . 100 g body wt-1 caused much higher increases in the biliary excretion of these proteins compared with the respective values in rats that received an infusion of TC at a threefold higher rate (1.2 mumol . min-1 . 100 g body wt-1). In contrast, a TUDC infusion at 1.8 mumol . min-1 . 100 g body wt-1 showed the minimum effect on these protein leakages. A combined infusion of TCDC (0.4 mumol . min-1 . 100 g-1) and TUDC (0.6 mumol . min-1 . 100 g-1) resulted in drastic (8- to 20-fold) decreases in excretion of these enzymes and albumin compared with respective values in rats infused with TCDC alone. Similar preventive effects were observed with the addition of TUDC to the infusion of TC (1.2 mumol . min-1 . 100 g-1). These results suggest that the hepatic cytotoxic effects of TC and TCDC can be prevented by the simultaneous infusion of TUDC in rats. PMID- 2984944 TI - Interaction of human lactoferrin with the rat liver. AB - Binding of human lactoferrin (hLf) by purified rat liver plasma membranes was studied to clarify whether the liver possesses specific hLf receptors. The binding was rapid between 4 degrees and 37 degrees C, with a pH optimum close to 5.0. At 22 degrees C and in glycine-NaOH (5 mM, pH 7.4) containing 150 mM NaCl and 0.5% albumin, 1 microgram of membrane bound a maximum of 11.8 ng hLf. The dissociation constant of the interaction was 1.6 X 10(-7) M. Other proteins of high isoelectric points (lactoperoxidase, lysozyme, and particularly salmine sulfate) and a piperazine derivative inhibited hLf binding in a concentration dependent manner. In contrast, monosaccharides (galactose, N-acetylgalactosamine, mannose, and fucose) were ineffective. By omitting NaCl from the incubation buffer, binding was increased 3.6-fold. Erythrocyte ghosts bound hLf less firmly and alveolar macrophages more firmly than hepatic plasma membranes. Liver cell fractionations performed after the intravenous injection of labeled hLf showed that approximately 88% of the hepatic radioligand was associated with parenchymal cells. When binding was expressed per unit of cell volume, however, more hLf was present in nonparenchymal than in parenchymal cells, implying that the above value was determined by the relative cell masses rather than affinities alone. It is concluded that the binding of hLf by hepatic plasma membranes is electrostatic, i.e., is mediated by the cationic nature of the ligand, and that it is explicable in terms of a "specific nonreceptor interaction" of the generalized type proposed by Cuatrecasas and Hollenberg (Adv. Protein Chem. 30: 251-451, 1976). PMID- 2984945 TI - Glucocorticoid effects on Na-K-ATPase in rabbit nephron segments. AB - We determined the effect of dexamethasone on Na-K-ATPase activity in six nephron segments of the adrenalectomized rabbit. Treatment consisted of 1.4 micrograms dexamethasone X 100 g body wt-1 X day-1 for 7 days prior to the study of the nephron segments. Enzyme activity was determined in individual nephron segments by a microfluorometric assay. There was 40-50% less activity of Na-K-ATPase in the S1 portion of the proximal convoluted tubule (PCT, S1), the medullary thick ascending limb (MTAL), and the distal convoluted tubule (DCT) of adrenalectomized rabbits compared with that of control (sham-operated) animals. There was no significant difference in the enzyme activity in proximal straight tubules (PST, S2 and S3) and cortical thick ascending limb (CTAL) of adrenalectomized and control animals. Dexamethasone treatment produced a dexamethasone concentration of 5 +/- 0.8 nM in the plasma and increased Na-K-ATPase activity in PCT (S1), MTAL, and DCT of the adrenalectomized animals to the control levels without significantly affecting the enzyme activity in the PST (S2, S3) or CTAL. The concentration of dexamethasone in the plasma was such that the hormone should bind mainly to dexamethasone receptors (Kd = 5 nM) and very little to aldosterone receptors (Kd greater than 60 nM). Thus, glucocorticoids probably stimulate Na-K ATPase in PCT, MTAL, and DCT through glucocorticoid (Type II) receptors and not through mineralocorticoid (Type I) receptors. PMID- 2984946 TI - Na-K-ATPase in isolated rabbit tubules after unilateral nephrectomy and Na+ loading. AB - Na-K-ATPase activity was studied in tubule segments from the cortex and medulla of rabbit kidneys under normal conditions, after unilateral nephrectomy, and after chronic salt loading. After unilateral nephrectomy kidney weight increased by 37% and Na-K-ATPase activity rose significantly in all nephron segments by 36 200% (P less than 0.01). Oral salt loading for 2 wk with 0.5% NaCl caused an increase in GFR and in absolute sodium excretion as well as reabsorption; plasma aldosterone decreased by 44% (P less than 0.005). In the proximal segments (PCT, CPST, OMPST, and TDL) there were no marked changes in Na-K-ATPase activity, whereas along the whole length of the ascending limb of Henle's loop (iMTAL, MTAL, and CTAL) there was a significant rise in the enzymatic activity of 30-200% (P less than 0.02). In the distal segments (DCT, CCD, and OMCD) there was a marked decrease of 50-60% (P less than 0.005) in Na-K-ATPase activity after the salt loading. We conclude that unilateral nephrectomy caused a general increase in Na-K-ATPase activity along the whole length of the nephron, and salt loading caused a selective increase in enzyme activity along the ascending limb of Henle's loop and decrease in the distal segments. PMID- 2984947 TI - Anion transport inhibitors: effects on water and sodium transport in the toad urinary bladder. AB - Acidification of the medium bathing the serosal surface of the toad urinary bladder results in impairment of the water permeability response to vasopressin. The magnitude of the hydrosmotic response to a maximal concentration of either vasopressin or the cyclic nucleotide analogue 8-(p-chlorophenylthio)-cyclic 3',5' adenosine monophosphate (C1PhS-cAMP) was progressively reduced when serosal bath pH was decreased from 8.5 to 6.5. The disulfonic stilbenes SITS and DIDS and the diuretic furosemide, agents known to interfere with anion transport and with the regulation of intracellular pH in other tissues, inhibited the water flow response to vasopressin and C1PhS-cAMP in a pH-dependent manner when added to the serosal bathing medium. Inhibition of the hydrosmotic response to 10(-5) M C1PhS cAMP was estimated to be half-maximal at 1.5 X 10(-4) M SITS, 2 X 10(-5) M DIDS, and 1 X 10(-5) M furosemide. The degree of inhibition induced by the anion transport inhibitors varied inversely with the concentration of exogenous cyclic nucleotide. SITS, DIDS, and furosemide had no effect on either basal or vasopressin-stimulated short-circuit current at serosal pH 8.5; all three agents inhibited basal short-circuit current at pH 7.1 but had no effect on the natriferic response to vasopressin. These results are consistent with the view that changes in intracellular hydrogen ion and/or anion concentration can selectively inhibit the increase in water permeability elicited by vasopressin at a step(s) distal to the generation of cAMP. PMID- 2984948 TI - Adrenal corticoids in hamsters: role in circadian timing. AB - The 24-h patterns of circulating cortisol and corticosterone were determined in male hamsters housed under a 14:10 light-dark cycle. Corticoid levels varied significantly over the 24-h sampling period with peak levels of both hormones occurring near the onset of the daily dark phase. The ratio of cortisol to corticosterone changed dramatically during the day. Corticosterone levels were significantly higher than cortisol during the early part of the light phase; however, cortisol levels became significantly higher than corticosterone when both hormones began their daily rise. To examine whether the circadian rhythm of cortisol secretion could be involved in the physiological control of hamster circadian organization, cortisol was infused at approximately physiological levels into adrenalectomized hamsters either continuously or in a 24-h rhythm. No significant differences were observed in the timing of circadian wheel-running rhythms in hamsters housed in LD 16:8, LD 14:10, or LL when cortisol was infused continuously, in a 24-h rhythm that mimicked the cortisol rhythm of intact hamsters, or in a 24-h rhythm several hours out of phase with the rhythm of intact hamsters. Provision of cortisol in a 24-h rhythm appeared to promote the survival of adrenalectomized hamsters since hamsters receiving a 24-h pattern of cortisol survived the experimental protocol significantly longer than those receiving the same dose of cortisol continuously. PMID- 2984949 TI - Infant formulas and gastrointestinal illness. AB - Infants under age one in a pediatric practice were followed prospectively, and the determinants of acute gastrointestinal illness were evaluated in case-control pairs, matched by birth month. The risk of acute gastrointestinal illness in infants receiving formula was six times greater than in infants receiving breast milk and 2.5 times greater than in infants receiving cow milk. In the second six months of life, infants on formula had 0.38 more gastrointestinal illness episodes per child than infants on cow milk. Episodes without rotavirus or bacterial agents accounted for most of the increased risk of formula. The increased risk could not be explained by iron fortification of the formulas, prescription of non-milk based formulas to high-risk infants, case ascertainment bias, control selection bias, or numerous control factors. Non-antibody anti infection properties found in cow milk are one possible explanation for these findings. PMID- 2984950 TI - In vitro culture of two populations (dividing and nondividing) of exoerythrocytic parasites of Plasmodium vivax. AB - Plasmodium vivax sporozoites invaded human hepatoma cells and differentiated into 2 types of exoerythrocytic (EE) parasites. One group was composed of actively dividing schizonts, which released merozoites after 9 days of culture. The second group was nondividing and persisted after the primary schizonts disappeared from the culture. EE schizonts progressively lost reactivity to monoclonal antibodies to the surface-protective protein antigen of P. vivax sporozoites, but the persisting parasites remained strongly reactive. In the 2 strains of P. vivax studied, the ratio of schizonts to persisting parasites was approximately equal. PMID- 2984952 TI - Blood transfusions and survival after lung cancer resection. AB - The immunosuppressive effects of blood transfusions in renal transplantation patients are now well documented. The question arises as to whether the possible immunosuppressive effects of blood transfusions in cancer patients cause a more favorable host environment for tumor growth. One hundred fifty-five patients undergoing resection for lung carcinoma were analyzed retrospectively, and it was shown that the use of blood transfusions was associated with a significant decrease in survival time in patients undergoing curative resection of lung carcinoma despite multivariate adjustments for age, sex, cell type, right lung versus left lung location, type of operation, and stage. This association supports, but does not prove, the hypothesis that blood transfusions, possibly through an immunosuppressive mechanism, are responsible for a poorer prognosis in patients who undergo resection for carcinoma of the lung. PMID- 2984951 TI - The isolation of arboviruses including a new flavivirus and a new Bunyavirus from Ixodes (Ceratixodes) uriae (Ixodoidea: Ixodidae) collected at Macquarie Island, Australia, 1975-1979. AB - Pools of ticks, Ixodes (Ceratixodes) uriae collected between 1975 and 1979 at Macquarie Island, yielded 33 strains of at least 4 different viruses: Nugget virus (Kemerovo group), 1 strain; Taggert virus (Sakhalin group) 9 strains; a previously undescribed flavivirus, related to Central European Tickborne encephalitis virus, for which the name "Gadgets Gully" is proposed, 9 strains; a virus serologically related to the Uukuniemi serogroup, family Bunyaviridae, for which the name "Precarious Point" is proposed, 10 strains. Three isolates were mixtures of Nugget and Gadgets Gully viruses; the remaining virus strain remains unidentified. PMID- 2984953 TI - [Neurosurgical collaboration in the treatment of various juvenile hemorrhagic fibroangiomas]. PMID- 2984954 TI - Microdetermination of phosphoinositides in a single extract. AB - A method that allows the quantification of phosphatidylinositol (PI), phosphatidylinositol 4-phosphate (DPI), and phosphatidylinositol 4,5-biphosphate (TPI) on a nanomolar scale is presented. The method is based on the simultaneous separation of lipids on high-performance thin-layer chromatography plates, followed by a microassay for phosphorus of PI spots and a densitometric assay of DPI and TPI. The new procedure allows the determination of the phospholipids in small amounts (100 micrograms protein) of synaptosomes and synaptic plasma membranes, and in homogenates of microwave-fixed brain tissue (1 mg wet wt). The usefulness of the method is illustrated by showing the effect of Ca2+ on the breakdown of DPI and TPI in synaptosomal plasma membranes. PMID- 2984956 TI - Nonrandom DNA sequencing of exonuclease III-deleted complementary DNA. AB - The nonrandom DNA sequence analysis procedure of Poncz et al. [Proc. Natl. Acad. Sci. USA 79, 4298-4302 (1982)] was extensively modified to permit the determination of complementary DNA (cDNA) sequences containing G-C homopolymer regions. The recombinant cDNA plasmid was cleaved at a unique restriction enzyme site close to the cDNA and treated with Exonuclease III under controlled conditions to generate a set of overlapping fragments having deletions 50-1500 bases in length at the free 3' termini. After removal of single-stranded DNA regions by Bal31 and DNA polymerase I large fragment, the unique restriction enzyme site was recreated by blunt end ligation of synthetic oligonucleotides to the deleted DNA fragments and restriction enzyme digestion. The cDNA fragment was excised from the cloning vector using a second different restriction enzyme having a unique site that flanks the cDNA fragment and subsequently force-cloned into either M13 mp10 or mp11. This method should also be particularly useful for the sequencing of other types of DNA molecules with lengths 1500 bp or smaller. PMID- 2984955 TI - Protein blotting: detection of proteins with colloidal gold, and of glycoproteins and lectins with biotin-conjugated and enzyme probes. AB - Methods to detect "native" proteins immobilized on nitrocellulose membranes in spot tests or on blots prepared from polyacrylamide slab gels after electrophoretic separation are described. Gold sols were found to be useful as general stains for proteins: They are polychromatic, yield an indelible record, and are complementary to india ink as protein stains because these two stains have different sensitivities for a number of proteins tested. For detection of wheat germ lectin (WGL)-binding glycoproteins, avidin-peroxidase was an effective enzyme probe, because the glycoportion of the avidin moiety possesses binding affinity to WGL. Glycocomponents in human parotid saliva were detected with this probe and with the following biotin-conjugated lectins as intermediary probes: soybean lectin, Bandeiraea simplicifolia lectin, Lotus tetragonolobus lectin, and kidney bean lectin. Autoclaving blots prior to probing eliminated endogenous peroxidase activity. Concanavalin A and WGL were separated by isoelectric focusing and detected on blots with horseradish peroxidase and avidin-peroxidase, respectively. The versatility of the biotin/avidin system was used to detect other lectins on similar blots using biotin-conjugated glycoproteins as intermediary probes: Helix pomatia lectin and B. simplicifolia lectin were detected with biotinyl neoglycoproteins, and kidney bean lectin with biotin conjugated components of parotid saliva. PMID- 2984957 TI - Assay of diadenosine tetraphosphate hydrolytic enzymes by boronate chromatography. AB - A new procedure was described for assay of diadenosine tetraphosphate (Ap4A) hydrolases based on boronate chromatography. Potential reaction products, AMP, ADP, and ATP, of the hydrolysis of Ap4A were separated from residual substrate by chromatography on a boronate-derivatized cation-exchange resin, Bio-Rex 70. Separation was achieved by changing the concentrations of ethanol and ammonium acetate in the elution buffers. Picomole masses of products were detectable, blank dpm values were less than 0.5% of the total dpm, and auxiliary enzymes were not required. The procedure was specifically described for Ap4A pyrophosphohydrolase from Physarum polycephalum. The assay is generally applicable for dinucleoside polyphosphate hydrolases which hydrolyze other substrates such as Ap3A, Ap5A, Ap6A, and Gp4G. Dinucleotide polyphosphates are readily purified by chromatography on this boronate resin in a volatile buffer. Tes, Tricine, and Tris buffers significantly interfered with the chromatography of ATP. PMID- 2984958 TI - Method for the preparation of human erythrocyte membrane with low basal calcium ATPase, responsive to stimulation. AB - A simple procedure for preparing erythrocyte membranes with low basal Ca2+ ATPase activity is described, which is stimulated several-fold by the addition of hemolysate in the incubation mixture. The cells are hemolyzed in hypotonic imidazole buffer and resulting membranes are washed with hypotonic phosphate buffer (pH 8.0) and the hemolyzing medium. The membrane preparations also have Mg2+-stimulated and Na+-K+-stimulated ATPase activities. The method allows the comparison of basal Ca2+ ATPase as well as hemolysate- or calmodulin-stimulated Ca2+ ATPase activities and thus may be useful in studying Ca2+ ATPase activity in various physiopathological conditions. PMID- 2984959 TI - Determination of standard potentials and electron-transfer rates for halobiphenyls from electrocatalytic data. PMID- 2984960 TI - Distribution of the neurons of origin of the great cerebral commissures in the cat. AB - Large injections of horseradish peroxidase throughout major portions of the right cerebral hemispheres of four cats revealed extensive distributions of the neurons of origin of the corpus callosum, the anterior commissure and the hippocampal commissures in the uninjected left hemispheres. The distributions of labelled neurons were mapped by semiautomatic computer microscope. The radial and tangential neuron distributions presented here are of a higher density and greater extent than those in previously published studies based on injections of transportable label to more circumscribed areas of the cerebral cortex of the cat. Generally, commissural neurons in the cat were distributed in a bilaminar fashion with supragranular cells more numerous than infragranular cells. PMID- 2984961 TI - Ionized calcium, the heart, and hemodynamic function. PMID- 2984962 TI - The antidotal action of thiosulfate following acute nitroprusside infusion in dogs. AB - The authors previously demonstrated in dogs that a bolus dose of sodium thiosulfate maintained enhanced cyanide metabolism throughout a 1-h infusion of sodium nitroprusside (SNP). To further test this antidotal action, a bolus dose of thiosulfate (150 mg . kg-1) was given to eight dogs at the end of a 60-min near-lethal infusion of nitroprusside (3 mg . kg-1). Within 2 min of the antidote, mean plasma thiocyanate levels (70.3 mumol . l-1) were significantly higher than those of seven control dogs given nitroprusside only (45.9 mumol . l 1, P = 0.002) and plateaued at 153.8 mumol . l-1 within 60 min, while the control values only reached 79.1 mumol . l-1 (P less than 0.001). Although differences between plasma cyanide levels in the two groups only attained significance 1 h after administering the antidote (0.8 vs. 2.74 mumol . l-1, P = 0.03), red blood cell cyanide concentrations were significantly lower in the antidote group within 5 min (166 vs. 225 mumol . l-1, P = 0.004) and remained so throughout the 2-h observation period. Compared with the controls, there was an impressive reduction in mean half-lives of plasma cyanide (25.1 vs. 74.1 min) and red blood cell cyanide (22.4 vs. 203.6 min). Similarly, peak cyanide levels occurred much sooner following the antidote (mean times: plasma cyanide 2.9 vs. 5.9 min; red blood cell cyanide 0.25 vs. 11 min).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984963 TI - Acute adrenal insufficiency in a patient with appendicitis during anesthesia. PMID- 2984964 TI - Increased serum IgG4 levels in acute Epstein-Barr viral mononucleosis. AB - Changes in serum IgG4 levels during heterophil-positive infectious mononucleosis were recorded. Pre-illness serial acute and convalescent specimens were evaluated. Mean IgG4 levels increased by 75% during acute infectious mononucleosis, reached a peak 21/2 to 3 weeks post-onset, and declined to baseline by 60 days. IgG4 peaked independently of other immunoglobulin classes. PMID- 2984965 TI - [Acute adult respiratory distress syndrome after lymphography]. AB - Pulmonary complications of lymphography are usually described as radiological infiltrates without clinical symptoms. However, a case is here reported of an adult respiratory distress syndrome occurring after lymphography in a 60 year old female lymphoma patient. Pulmonary oedema developed within 48 h; haemodynamic study showed a normal capillary wedge pressure. The patient died from intractable low cardiac output within 24 h. Post-mortem examination showed pulmonary lymphocytic infiltration and multiple fat emboli. The lack of lymphatic drainage was probably responsible for the intravascular passage of lipid-soluble contrast medium, this giving endothelial lesions. In such patients with preexisting lung disease or pulmonary involvement in haematological disease, lymphography has to be considered carefully. PMID- 2984966 TI - Prolonged cardiac arrest and resuscitation in dogs: brain mitochondrial function with different artificial perfusion methods. AB - Clinical techniques for artificial perfusion have not previously been examined directly for their effects on brain high-energy metabolism. Our study involved 24 large mongrel dogs that were anesthetized, instrumented for central venous intravenous access, and subjected to craniotomy to expose the dura and underlying parietal cortex. The animals were divided into the following six experimental groups of four animals each: nonischemic controls; 15-minute cardiac arrest without resuscitation; 45-minute cardiac arrest without resuscitation; 15-minute cardiac arrest plus 30 minutes resuscitation with conventional cardiopulmonary resuscitation (CPR); 15-minute cardiac arrest plus 30 minutes resuscitation with interposed abdominal compression (IAC) CPR; and 15-minute cardiac arrest plus 30 minutes resuscitation with internal cardiac massage. Cardiac arrest was induced by central venous injection of KCl 0.6 mEq/kg, and it was confirmed by continuous ECG monitoring. The three active resuscitation models included administration of NaHCO3 and epinephrine, but no attempt was made to restart the heart by defibrillation during resuscitation. At the indicated time in each group, a 4- to 5-g sample of brain was removed through the craniotomy, immediately cooled to 0 C and processed for isolation of mitochondria. The mitochondria were studied for their content of superoxide dismutase and for quantitative oxygen consumption with glutamate/malate substrate during resting and ADP-stimulated respiration. Our results show a significant drop in brain mitochondrial superoxide dismutase activity during the first 15 minutes of cardiac arrest. There is minimal injury to brain mitochondrial oxygen consumption during both 15 and 45 minutes of complete ischemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2984967 TI - [Reye's syndrome and cytomegalovirus]. PMID- 2984968 TI - [Hemolytic anemia as a complication of infectious mononucleosis]. PMID- 2984969 TI - [Postinfection acquired immunohemolytic anemia in children]. PMID- 2984970 TI - [Vitamin D3 poisoning and irreversible sequela]. AB - Twenty-four children with vitamin D intoxication and a follow-up of one to thirteen years old (means: four years and seven months) are reviewed. Over-dosage was prescribed by medical order in 66.6% of patients and by the mother herself in 16.6%. Intensity of clinical symptoms (renal, neurologic, digestive) were related with daily dose administered whilst final secuelae depends on duration of overdosage. Hipercalcemia was easily corrected by association of low calcium diet, corticoesteroids and/or furosemide in least than a month in 81% of cases. Two patients died during the acute fase and 22.7% remain with permanent damage (five in chronic renal failure, one in haemodialysis and three with low IC). PMID- 2984971 TI - Hematologic malignancies associated with primary mediastinal germ-cell tumors. AB - Three men with primary mediastinal germ-cell tumors subsequently developed a malignant hematologic disorder characterized by pancytopenia and marrow infiltration with hematopoietic blast cells. Two of these patients were classified as having acute megakaryocytic leukemia and the third was believed to have a myelodysplastic syndrome with a prominent megakaryocytic component. Analysis of clinical characteristics of these patients and review of the literature suggest that the proximate association of mediastinal germ-cell tumors with malignant hematologic disorders is neither a coincidence nor a consequence of chemotherapy given for the germ-cell tumor. We believe this association represents the evolution of a neoplastic disorder that initially involves a totipotent germ cell. These germ cells, when located in the mediastinum, apparently acquire hematologic phenotypes and are manifested clinically as a hematologic malignancy. PMID- 2984972 TI - Antibody to human T-lymphotropic virus type III in wives of hemophiliacs. Evidence for heterosexual transmission. AB - To evaluate the risk of heterosexual transmission of the acquired immunodeficiency syndrome, lymphadenopathy, and infection with human T lymphotropic virus type III (HTLV-III), we studied 42 hemophiliacs and their wives. By early 1984, 9 of the hemophiliacs had asymptomatic lymphadenopathy and 1 had the acquired immunodeficiency syndrome. Twenty-one hemophiliacs, including all 10 with clinically overt disease, had antibody to HTLV-III. None of the 42 wives had lymphadenopathy or the acquired immunodeficiency syndrome but 2 had HTLV-III antibody. One of these women had evidence of immunologic dysfunction with a markedly reduced T-helper/suppressor cell ratio. The husbands of these 2 women both had HTLV-III antibody, but neither had overt acquired immunodeficiency syndrome-related disease. Thus, as of early 1984, the prevalence of HTLV-III antibody in wives of hemophiliacs seropositive for HTLV-III was 9.5% (2 of 21). We conclude that transmission of HTLV-III occurs between hemophiliacs and their heterosexual partners. PMID- 2984974 TI - Epstein-Barr virus and unexplained illness. PMID- 2984973 TI - Persistent infection with human T-lymphotropic virus type III/lymphadenopathy associated virus in apparently healthy homosexual men. PMID- 2984975 TI - Ultrasound in monitoring patients with hepatocellular carcinoma treated with adriamycin. PMID- 2984976 TI - Percutaneous irradiation in cholangio-carcinoma. PMID- 2984978 TI - Central pattern generators for locomotion, with special reference to vertebrates. PMID- 2984977 TI - Neuropeptide function: the invertebrate contribution. AB - The following is a list of generalizations that arise from considering the present state of knowledge concerning the functions of invertebrate peptides. Some of these clearly also apply to vertebrates. Invertebrate peptides can be classified into structurally related groups. Structural similarity of peptides may represent true evolutionary homology by selection acting on an original gene. Alternatively, independent evolution of similar genes may have occurred because certain amino acid sequences represent optimal solutions to complex functional problems. Invertebrate neuropeptides have multiple functions. Thus, proctolin is a cardioactive peptide, a skeletal neuromuscular transmitter, a hindgut neuropeptide, a peptide of CNS interneurons (Keshishian & O'Shea 1984) and may have humoral roles. Invertebrate peptides act through a variety of molecular mechanisms. Generalizations about the mechanism cannot yet be made. Thus, proctolin's action on crustacean skeletal muscle is not associated with stimulation of cyclic-AMP or protein phosphorylation, but the action of SCPB on molluscan skeletal muscle involves elevation of cAMP. Invertebrate peptide inactivation can be caused by proteolysis that can also function to enhance peptide bioactivity. Proctolin is made virtually biologically inactive by any proteolysis, but alpha-BCP bioactivity is enhanced by two steps of carboxy peptidase digestion before being functionally inactivated. Protease action on released peptides is not necessarily a "simple" form of transmitter inactivation. Protease action also involves functional processing whereby the temporal and spatial parameters of a peptide's action may be shaped. Invertebrate neuropeptides are frequently co-localized with other neuroeffectors. Peptides may be co-localized and released with other active peptides as in the bag-cell example, or may be coactive with more conventional transmitters, as in the Ds motoneuron example. In such circumstances there is no reason to view either transmitter as primary or secondary. Invertebrate neuropeptides are widely involved in the control of muscle contraction. These effects may be locally and directly mediated as in the Ds motoneuron example or may be humoral. The peptide may act directly on the muscle contractile system or function to modulate the muscles' response to other motor input. Muscle contraction may be induced by a neuropeptide without depolarization of the muscle cells, for example see proctolin. Invertebrate neuropeptides are frequently involved in the control of oscillatory functions. In several examples peptides activate rhythmic myogenic contractions of cardiac and skeletal muscle.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2984979 TI - Possible applications of recombinant DNA technology to the diagnosis and study of variants of alpha 1-antitrypsin. AB - We describe the basic principles of recombinant DNA technology and how this methodology has been used to isolate and characterise cloned genes coding for alpha 1-antitrypsin. We show how these probes can be used for diagnosis and to study molecular variants of alpha 1-antitrypsin which may predispose individuals to develop lung disease. PMID- 2984980 TI - Controlled trial of enviroxime against natural rhinovirus infections in a community. AB - The therapeutic effect of intranasally administered enviroxime was tested against naturally occurring common colds. The double-blind evaluation was carried out in Tecumseh, Mich., during a period when rhinoviruses are usually the principal pathogen. Rhinovirus transmission followed the typical pattern during this period of study. Although there were trends indicating greater therapeutic effectiveness for enviroxime when certain nasal symptoms were considered, there were no consistent statistically significant differences between treated and untreated groups. Results were unchanged when illnesses in different periods or associated with rhinovirus isolation were examined. It was concluded that no therapeutic effect of enviroxime was demonstrated. PMID- 2984981 TI - Survey of plasmids and resistance factors in Campylobacter jejuni and Campylobacter coli. AB - A total of 688 isolates of Campylobacter jejuni and Campylobacter coli were screened for the presence of plasmid DNA by agarose gel electrophoresis and were tested for susceptibility to ampicillin, chloramphenicol, erythromycin, streptomycin, and tetracycline. Of the isolates examined, 32% were noted to harbor plasmid DNA, ranging in size from 2.0 to 162 kilobases. Only tetracycline resistance was noted to correlate with the presence of plasmids. Plasmids capable of transferring tetracycline resistance via conjugation ranged in size from 42 to 100 kilobases. The Bg/II and Bc/I restriction endonuclease profiles of 31 plasmids examined showed marked diversity in their banding patterns. Although a high degree of DNA-DNA homology was noted among the Campylobacter spp. plasmids, no homology was noted between these plasmids and tetracycline R factors commonly found in the family Enterobacteriaceae. PMID- 2984982 TI - Duration of effect of interferon aerosol prophylaxis of vesicular stomatitis virus infection in mice. AB - Mice were exposed for 8 h to continuous small-particle aerosols containing natural mouse alpha interferon (estimated dosage 100 U per mouse) or one of two concentrations of hybrid recombinant alpha interferon A/D bgl (estimated dosages of 100 and 10,000 U per mouse, respectively). On days 1, 3, 5, 7, and 9 after exposure to these interferons, three mice from each group were inoculated intranasally with 100 PFU of vesicular stomatitis virus. Control mice were exposed to aerosols of saline or inoculated intraperitoneally with either natural mouse alpha interferon (350 U) or one of two doses of hybrid recombinant alpha interferon A/D bgl (350 or 35,000 U) and challenged similarly. Of mice injected intraperitoneally, only those given 35,000 U of hybrid recombinant alpha interferon A/D bgl 24 h before virus challenge were protected from pulmonary infection, compared with the saline-treated control mice. Of mice given 100 U of either interferon by small-particle aerosol, only those exposed 24 h before inoculation of vesicular stomatitis virus had reduced pulmonary titers of the virus. However, of mice given ca. 10,000 U of hybrid recombinant alpha interferon A/D bgl by small-particle aerosol, all groups except those exposed 9 days before virus inoculation had significantly reduced lung virus titers. PMID- 2984983 TI - Antibacterial effect of lactoperoxidase and myeloperoxidase against Bacillus cereus. AB - An oral periodontopathic bacterium, Bacillus cereus, was inhibited both by lactoperoxidase (LP) and myeloperoxidase (MP) antimicrobial systems. With the LP SCN--H2O2 system, the growth inhibition was directly proportional to the amount of OSCN- ions present. The OSCN-, which is the principal oxidation product of the LP (or MP)-SCN--H2O2 system at neutral pH, is a normal component of human saliva. The oxidation products of both peroxidase systems inhibited the growth of the bacteria. This inhibition was associated with reduced extracellular release of collagenase activity from the cells. With LP, the antimicrobial efficiency of the oxidizable substrates was SCN- greater than I-, and with MP, the efficiency was I greater than Cl- greater than SCN-, respectively. LP did not oxidize Cl-. PMID- 2984984 TI - Oral acyclovir therapy of genital herpes simplex virus type 2 infections in guinea pigs. AB - Oral acyclovir was evaluated for its effectiveness in treating guinea pigs with primary herpes simplex virus type 2 infections. Guinea pigs inoculated intravaginally with acyclovir-susceptible strains (for which 50% inhibitory concentrations of acyclovir in cell culture were found to be in the range of 0.15 to 1.2 micrograms/ml) and treated with 5.0 mg of acyclovir per ml in the drinking water beginning 48 h postinfection showed significant reductions in lesion severity. This dosage produced serum acyclovir levels of 1.3 micrograms/ml. Lower concentrations of oral acyclovir (less than or equal to 2.5 mg/ml in the drinking water), which produced serum acyclovir levels of less than 1.0 microgram/ml, were less consistently effective against these same virus strains. When an acyclovir resistant isolate (for which the 50% inhibitory concentration of acyclovir in cell culture was found to be 8.5 micrograms/ml) was used to initiate infection, treatment with 5 or 10 mg/ml (yielding serum levels of 1.3 and 3.5 micrograms/ml) in the drinking water had only minimal clinical benefit. However, the degree of response was difficult to determine because of the attenuated disease produced by the acyclovir-resistant virus. In vitro virus sensitivity may be predictive of the serum drug levels that need to be obtained to produce a successful response to therapy. PMID- 2984986 TI - Relatedness of tetracycline resistance plasmids among species of coagulase negative staphylococci. AB - Four isolates of Staphylococcus aureus and 98 isolates of coagulase-negative staphylococci representing six species all obtained from endocervical cultures were examined for antimicrobial susceptibility and for the presence of plasmids. More than 80% of the isolates were susceptible to each of 12 antimicrobial agents tested, whereas only 33% were susceptible to penicillin G, 30% were susceptible to cadmium chloride, and 41% were susceptible to tetracycline. Although no species-related susceptibility or plasmid patterns were detected, 77 isolates contained at least one plasmid and 43 contained a plasmid similar in mass to a 2.7-megadalton tetracycline resistance plasmid previously reported in staphylococci. Association of tetracycline resistance with plasmids of this size in four species was determined from curing experiments. No plasmids homologous with the tetracycline resistance locus of the Escherichia coli plasmid pBR322 were found among 11 isolates examined by DNA hybridization. Homology with a 2.7 megadalton plasmids (pRC701) from an endocervical isolate of S. aureus, however, was apparent for 2.7-megadalton plasmids harbored by six isolates as well as with larger plasmids harbored by three isolates. Restriction analysis revealed that pRC701 shared structural identity with two plasmids of a similar mass from two species of coagulase-negative staphylococci as well as with a previously characterized tetracycline resistance plasmid originating in S. aureus. PMID- 2984985 TI - Structure-activity studies on phosphonoacetate. AB - Phosphonoacetic acid is a selective antiherpesvirus agent. More than 100 congeners of phosphonoacetic acid were evaluated in vitro and in vivo to understand structure-activity relationships in the hope of designing a superior analog. Results showed that the antiherpesvirus activity had highly specific structural requirements. Neither the carboxylic nor the phosphono groups could be replaced. The distance between these two groups is important. Increase of this distance caused complete loss of activity. However, if this distance was maintained, the addition of groups to the methylene carbon resulted in a reduction, but not loss, of activity. On the other hand, decrease of the carbon chain to formic acid did not deteriorate its antiherpes activity. All analogs tested had lower activity than the parent compound. However, some compounds with decreased activity in vitro appeared to have favorable pharmacological properties in vivo. PMID- 2984987 TI - Distinctive properties of DNA polymerases induced by herpes simplex virus type-1 and Epstein-Barr virus. AB - The properties of DNA polymerases induced by two human herpesviruses, herpes simplex virus type-1 (HSV-1) and Epstein-Barr virus (EBV), have been compared. The HSV-1 and EBV polymerases can be distinguished from one another by differences in the elution profiles in phosphocellulose and single-stranded DNA cellulose columns. Although both enzymes require monovalent cations for optimum activity, the HSV-1 enzyme requires ammonium sulfate whereas the EBV enzyme activity is inhibited by it; on the other hand, the EBV polymerase requires KCl. Other reaction requirements are also different for the two viral enzymes. Thus, when the EBV DNA polymerase was assayed under conditions optimum for the HSV-1 DNA polymerase, only 15% of its activity was expressed. Differences were also noted in sensitivities of the two viral enzymes to the 5'-triphosphates of nucleoside analogs with antiherpesvirus activity such as BVdU, IVdU, ACV, FIAC and IdUrd. The HSV-1 polymerase was more sensitive than the EBV DNA polymerase to inhibition by phosphonoacetate, phosphonoformate, aphidicolin and N ethylmaleimide. However, the EBV DNA polymerase was more sensitive than HSV-1 DNA polymerase to heat treatment at 42 degrees C. Thus, the marked differences between the two viral enzymes can be useful in identifying enzyme activities in cells producing the virus and also in studying the biochemical mechanism of action of some of the antiviral agents. PMID- 2984988 TI - Combination chemotherapy: interaction of 5-methoxymethyldeoxyuridine with trifluorothymidine, phosphonoformate and acycloguanosine against herpes simplex viruses. AB - Methoxymethyldeoxyuridine (MMUdR) when used in combination with either trifluorothymidine (F3TdR) or phosphonoformate (PFA) showed synergistic activity against herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) in vitro, whereas MMUdR and acycloguanosine (ACG) combination was antagonistic against herpes viruses. HSV-1 mutants resistant to ACG, arabinofuranosyladenine (Ara-A), MMUdR or PFA were isolated. Drug-resistant HSV-1 virus mutants were analyzed for cross sensitivity to ACG, Ara-A, F3TdR, MMUdR, MMUdR-5'-monophosphate (MMUdR-MP) and PFA. The Ara-A-resistant (Ara-AR) virus exhibited 3-fold resistance to MMUdR MP (ID50 = 105 microM). The ACG-resistant (ACGR) mutant was 160-fold less sensitive to MMUdR (ID50 greater than 1138 microM). The MMUdR-resistant (MMUdRR) mutant remained sensitive to all other antiviral drugs in vitro. Ara-A provided protection against HSV-1 encephalitis in immunosuppressed mice inoculated with a low dose (200 PFU/mouse) of MMUdRR virus or wild-type HSV-1. F3TdR decreased incorporation of tritiated deoxyuridine [( 3H]UdR) in RK-13 cells by 50% at 0.068 microM. Under similar conditions, MMUdR (up to 600 microM) and PFA (up to 208 microM) were without effect on incorporation of [3H]UdR into DNA. In combination chemotherapy experiments, MMUdR (up to 300 microM) used along with F3TdR (up to 1.08 microM) neither decreased nor enhanced cytotoxicity of F3TdR as measured by incorporation of [3H]UdR into cellular DNA. Similarly, MMUdR (up to 300 microM) in combination with PFA (up to 166 microM) was nontoxic to host cells. PMID- 2984989 TI - Effects of humic and fulvic acids on poliovirus concentration from water by microporous filtration. AB - Because naturally occurring organic matter is thought to interfere with virus adsorption to microporous filters, humic and fulvic acids isolated from a highly colored, soft surface water were used as model organics in studies on poliovirus adsorption to and recovery from electropositive Virosorb 1MDS and electronegative Filterite filters. Solutions of activated carbon-treated tap water containing 3, 10, and 30-mg/liter concentrations of humic or fulvic acid were seeded with known amounts of poliovirus and processed with Virosorb 1MDS filters at pH 7.5 or Filterite filters at pH 3.5 (with and without 5 mM MgCl2). Organic acids caused appreciable reductions in virus adsorption and recovery efficiencies with both types of filter. Fulvic acid caused greater reductions in poliovirus recovery with Virosorb 1MDS filters than with Filterite filters. Fulvic acid interference with poliovirus recovery by Filterite filters was overcome by the presence of 5 mM MgCl2. Although humic acid reduced poliovirus recoveries by both types of filter, its greatest effect was on virus elution and recovery from Filterite filters. Single-particle analyses demonstrated MgCl2 enhancement of poliovirus association with both organic acids at pH 3.5. The mechanisms by which each organic acid reduced virus adsorption and recovery appeared to be different for each type of filter. PMID- 2984990 TI - Comparison of the airborne survival of calf rotavirus and poliovirus type 1 (Sabin) aerosolized as a mixture. AB - A mixture of a cell culture-adapted strain (C-486) of calf rotavirus and poliovirus type 1 (Sabin) was prepared in tryptose phosphate broth containing 0.1% uranine (physical tracer) and antifoam at a final concentration of 0.001%. By using a six-jet Collison nebulizer, the mixture was aerosolized into a 300 liter stainless-steel rotating (4 rpm) drum. The temperature of the air inside the drum was kept at 20 +/- 1 degrees C, and the virus aerosols were held at the following three levels of relative humidity (RH): low (30 +/- 5%), medium (50 +/- 5%), and high (80 +/- 5%). An all-glass impinger, containing 10.0 ml of tryptose phosphate broth with antifoam, was used to collect samples of air from the drum. Both viruses were propagated and quantitated in MA-104 cells. The calf rotavirus was found to survive well at mid-range RH, where 60% of the infectious virus could be detected even after 24 h of virus aerosolization. At the low RH, the half-life of the infectious rotavirus was ca. 14 h. On the other hand, no infectious poliovirus could be recovered from the drum air at the low and medium RH. At the high RH, more than 50% of the infectious rotavirus became undetectable within 90 min of aerosolization. In contrast to this, the half-life of the poliovirus at the high RH was about 10 h. These data, based on the aerosolization of virus mixtures, therefore suggest that there is a pronounced difference in the way RH influences the airborne survival of these two types of viruses held under identical experimental conditions. PMID- 2984991 TI - Virion conformational forms and the complex inactivation kinetics of echovirus by chlorine in water. AB - Aberrant inactivation kinetics were observed when monodispersed echovirus type 1 (Farouk) was inactivated with chlorine. An initial 1- to 2-log10-unit decrease in titer was followed by lag period, during which the titer stayed the same or increased, and this was followed by a final decline in titer. First-order kinetics were obtained with poliovirus type 1 under the same conditions. Isoelectric focusing studies of echovirus before chlorine treatment showed that the virus distributed into two pH-dependent and interconvertible isoelectric forms. After chlorine treatment all remaining virus infectivity was associated with a third pH-independent isoelectric form. The complex inactivation kinetics appeared to be due to shifts between these conformational forms during inactivation in certain ionic environments. Under certain conditions the conformational shifts resulted in substantial resistance of monodispersed echovirus to chlorine. PMID- 2984992 TI - Modified membrane-filter procedure for concentration of enteroviruses from tap water. AB - Enteroviruses added to 114 liters of dechlorinated tap water were recovered in a 16-ml sample by a two-stage concentration procedure in which different types of membrane filters were used in each concentration stage. Viruses in tap water at pH 3.5 were first adsorbed to 10-in. (ca. 25.4-cm) epoxy-fiber glass filters (Filterite). Viruses adsorbed to these filters were eluted with a solution of 0.2 M sodium trichloroacetate buffered at pH 9 with 0.2 M lysine. Viruses in this solution were adsorbed to 47-mm asbestos filters (Seitz) without pH adjustment or other modification of the solution. Viruses were recovered from the Seitz filters with 16 ml of either Casitone or fetal calf serum at pH 9. With these procedures ca. 45% of several types of enteroviruses added to 114 liters of tap water could be recovered in the final 16-ml sample. PMID- 2984993 TI - The appearance of 1,25-dihydroxyvitamin D3 receptor during chick embryo development. AB - The appearance of the 1,25-dihydroxyvitamin D3 receptor in intestine, kidney, and chorioallantoic membrane of chick embryo was followed by sucrose density gradient sedimentation analysis and Scatchard plot analysis. The receptor from each of these organs sediments as a single 3.7S component. At 19 days of embryonic life, intestine had the highest specific 1,25-dihydroxyvitamin D3 binding activity followed by kidney and chorioallantoic membrane. The 1,25-dihydroxyvitamin D3 binding activity increased gradually at 12-15 days and rapidly until 20 days in intestine. In kidney, this protein increased rapidly from 12 to 16 days and did not change subsequently. In chorioallantoic membrane, the receptor increased slowly from 8 through 15 days, rapidly until 19 days, and decreased at 20 days. The injection of hydrocortisone into the chick embryo at 10 days increased receptor number in intestine, kidney, and chorioallantoic membrane by a factor of 2 at 12 days. Injection of this hormone after this time had little or no effect. PMID- 2984994 TI - Effect of insulin secretagogues and potential modulators of secretion on a plasma membrane (Ca2+ + Mg2+)-ATPase activity in islets of Langerhans. AB - Studies were undertaken to determine whether factors which affect insulin secretion may exert their effects by altering the activity of an islet-cell plasma membrane Ca2+ extrusion pump. The insulin secretagogue, D-glucose, and a variety of phosphorylated hexoses, glucose 6-P, glucose 1,6-P, fructose 6-P, and fructose 2,6-P, were evaluated for their effect on an islet-cell plasma membrane (Ca2+ + Mg2+)-ATPase and were found to be ineffective in altering enzyme activity. D-Glucose also did not alter the rate of ATP-dependent Ca2+ uptake into plasma membrane vesicles. Similarly, cAMP, the catalytic subunit of cAMP dependent protein kinase, arachidonic acid, or prostaglandin E2 did not affect either the plasma membrane (Ca2+ + Mg2+)-ATPase or the rate of ATP-dependent Ca2+ uptake into plasma membrane vesicles. Whereas previous studies have suggested that D-glucose and/or cAMP may inhibit ATPase activities in islets, these results indicate that the agents, i.e., D-glucose and cAMP, which stimulate and/or potentiate insulin secretion from the islet cell, do not modify Ca2+ fluxes by directly regulating the islet-cell plasma membrane (Ca2+ + Mg2+)-ATPase. In contrast, the acidic phospholipids, phosphatidic acid and phosphatidylserine, stimulated the enzyme activity in a concentration-dependent manner whereas phosphatidylcholine had only a minimal effect. The diacylglycerol, dilinolein, stimulated the (Ca2+ + Mg2+)-ATPase activity in the presence of phosphatidylserine, but not in the absence of phospholipids. These effects were independent of phospholipid-stimulated protein phosphorylation in the islet-cell plasma membrane under the conditions of the ATPase assay. PMID- 2984995 TI - Localization of lysines acetylated in ubiquitin reacted with p-nitrophenyl acetate. AB - The protein ubiquitin undergoes extensive N epsilon-acetylation of some of its seven lysine residues when reacted with p-nitrophenyl acetate. Lysines 27 and 29 show little reactivity whereas residue 6 is the most readily acetylated. Residues 11, 33, 48, and 63 show intermediate reactivities. PMID- 2984996 TI - Direct effects of ethanol on bone resorption and formation in vitro. AB - In vitro studies indicate that low concentrations of ethanol can have direct effects on bone formation and resorption. Bone resorption was increased when embryonic chick tibiae were exposed to ethanol at 0.03-0.3% (v/v), and bone formation was inhibited when tibiae were exposed to 0.2% ethanol in the presence of NaF or parathyroid hormone (P less than 0.01 for each). Ethanol also had direct effects on isolated bone cells in vitro, increasing both cAMP and PGE2 production (P less than 0.001 for each), and affecting cell proliferation in a biphasic, time- and dose-dependent manner. After 24 h of exposure, 0.03% ethanol increased bone cell proliferation (P less than 0.001), but 0.3% ethanol was inhibitory (P less than 0.01). Paradoxically, mitogenic doses of ethanol prevented the effects of two other mitogens, NaF and human skeletal growth factor, to increase bone cell proliferation (P less than 0.001). But how were these effects produced? Several observations suggest that these direct effects of ethanol on skeletal tissues in vitro were mediated by changes in bone cell membrane fluidity. (a) Dimethyl sulfoxide, ethylene glycol, and lecithin, which act, like ethanol, to increase membrane fluidity, mimicked the effects of ethanol on bone cell proliferation. Dimethyl sulfoxide also mimicked the effect of ethanol to increase cAMP (P less than 0.001). (b) Cholesterol, which decreases cell membrane fluidity, acted oppositely to ethanol and enhanced the mitogenic response to human skeletal growth factor (P less than 0.001). (c) Preincubation of calvarial cells with ethanol or with cholesterol altered the in situ reaction kinetics of the membrane-bound enzyme, alkaline phosphatase. Together, these data demonstrate that ethanol has direct effects on skeletal tissue in vitro, and suggest that those effects may be secondary to changes in bone cell membrane fluidity. PMID- 2984997 TI - The presence of two heat-stable inhibitors of phosphoprotein phosphatases in the dimorphic fungus, Mucor rouxii. AB - Two heat-stable inhibitors (a and b) of phosphoprotein phosphatases I and II from Mucor rouxii were isolated from mycelium of the fungus. They were partially purified from extracts by heating, DEAE-cellulose chromatography, and Sephadex G 75 gel filtration. The molecular weights of inhibitors a and b, estimated by gel filtration, are 5,000 and 20,000 respectively. Inhibitor a acts similarly on both enzymes while inhibitor b is relatively more active on enzyme II. Storage of inhibitor b at -20 degrees C for several weeks resulted in a partial conversion to a lower-molecular-weight form with properties similar to those of inhibitor a. PMID- 2984998 TI - Properties of brain dolichol kinase activity solubilized with a zwitterionic detergent. AB - Dolichol kinase activity is effectively solubilized by extracting calf brain microsomes with 2% 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), a zwitterionic detergent. The solubilized kinase catalyzes the enzymatic phosphorylation of dolichols with either CTP or dCTP serving as phosphoryl donor in the presence of Ca2+. Similar Km values were calculated for CTP (7.7 microM) and dCTP (9.1 microM). Dolichol phosphorylation was inhibited by CDP and dCDP, but not CMP, ADP, GDP, or UDP. A kinetic analysis of the inhibitory effect of CDP revealed a pattern characteristic of competitive inhibition. Dolichol kinase activity was markedly stimulated by the addition of R-dolichol (C95) or S dolichol(C95). The apparent Km value for R-dolichol(C95) and S-dolichol(C95) was 9 microM, but the Vmax for the phosphorylation reaction was 40% higher with S dolichol(C95). Incubation of the CHAPS extract with [gamma-32P]CTP and exogenous undecaprenol(C55) resulted in the enzymatic synthesis of a radiolabeled product that was mild acid-labile and chromatographically identical to undecaprenyl monophosphate. An enzymatic comparison with a variety of polyprenol substrates indicates that the solubilized kinase prefers long-chain (C90-95) polyprenols with saturated alpha-isoprene units. The effect of exogenous phosphoglycerides on the kinase activity in the dialyzed CHAPS extracts has also been evaluated. These studies describe the properties and polyprenol specificity of stable, solubilized preparations of dolichol kinase that should be useful for further purification of the enzyme. PMID- 2984999 TI - [A member of the SRC gene family, the c-erbB-1 gene, is closely related to the EGF receptor gene]. AB - An avian erythroblastosis virus, AEV-H, induces both erythroblastosis and sarcomas in susceptible chickens. Since AEV-H carries the v-erbB as a sole oncogene, the erbB gene was suggested to be responsible for the induction of these tumors. Analysis of the amino acid sequence predicted from the nucleotide sequence of the v-erbB gene revealed that the gene product has a domain characteristic for tyrosine kinase. Recently in has been suggested has the v-erbB protein is a part of the chicken EGF (epidermal growth factor) receptor. Using antibody against either v-erbB protein or EGF receptor, we also demonstrated close similarity between the two proteins. Further studies on human genomic DNA revealed that the c-erbB-1 gene, a proto-oncogene of the v-erbB gene, is the EGF receptor gene. We were also able to identify the c-erbB-2 gene that seems to code for a EGF receptor-like protein with a domain for tyrosine kinase. Finally, we would like to show that cell lines established from human squamous cell carcinom are frequently associated with amplification of the c-erbB-1/EGF receptor gene. PMID- 2985000 TI - [Cellular responses and the H+ pump in the vacuolar system]. AB - Intracellular acidic compartments are now recognized to be ubiquitous in the vacuolar system of eukaryotic cells, where the unique ATP-dependent H+ pump (s) functions to maintain their internal low pH. Furthermore, various basic substances and acidic ionophores have been found to raise the internal pH of these compartments to neutrality. These chemicals, on the other hand, have been found to affect various biological reactions in multicellular organisms, including those involved in intercellular communication. The properties of the H+ pump of the vacuolar system and its role in cellular responses are described. PMID- 2985001 TI - [Somatic cell mutants with altered receptor and endocytosis activities]. AB - Binding and endocytosis are responsible for cellular uptake of a wide variety of physiologically active ligands. We have selected somatic cell mutants with altered endocytosis activity for low-density lipoprotein, insulin, and epidermal growth factor. Of these mutants, the one with altered response to epidermal growth factor shows aberrant growth behavior in soft agar. PMID- 2985002 TI - [Abnormal (constitutive) expression of interleukin 2 receptor (Tac Ag) in adult T cell leukemia cells]. PMID- 2985003 TI - [Mitogenic responsiveness to growth factor and tumor promoters and regulation of cell growth]. AB - Most of our current understanding of the molecular action of mitogens such as insulin, EGF and tumor promoters is derived from studies using biochemical, pharmacological and immunological methods. In the present study, however, this important problem has been approached from a genetic point of view. To facilitate genetic studies, attempts have been made to isolate variants having receptors and/or post-receptor functions which are deficient or extraordinary in their cellular responsiveness. The first selection method was to use toxic hybrid proteins such as insulin crosslinked to the A fragment of diphtheria toxin (DTa), EGF crosslinked to the A chain of toxic ricin (RICa) and monoclonal anti-EGF receptor antibody (IgG) crosslinked to the RICa. These conjugates have provided us with a variety of cell variants which are unable to either bind the ligand or take up the bound ligand and process it for mitogenic response. The second selection procedure utilized the mechanical removal of cells whose growth was stimulated by the mitogens but was arrested at mitosis by vinblastine sulphate. Variants of mouse 3T3-L1 cells which no longer responded to the indole alkaloid tumor promoter dihydroteleocidin B exhibited various characteristics in terms of the receptors for EGF, insulin and the tumor promoters. The results obtained from these cell variants are discussed in relation to the role of different receptor systems in the regulation of DNA replication, cell growth and differentiation. PMID- 2985004 TI - [Early cancer and related changes in the bronchial epithelium of former mustard gas workers]. AB - The bronchial epithelium taken in stepwise transverse sections was examined histologically in 66 autopsy cases, composed of groups consisting of 19 mustard gas (MG) ex-workers with lung cancer, 17 MG ex-workers with non-lung cancer, 10 non-MG lung cancer cases, and 20 non-MG non-lung cancer cases. An additional 5 surgical lung cancer specimens removed from MG ex-workers were also examined. From these groups, foci of moderate or severe atypia including cellular atypia, dysplasia and carcinoma in situ (CIS), detected in the total number of slides for each autopsy group, were counted as 146 out of 3,485, 72 out of 2,226, 70 out of 3,797, and 18 out of 4,611, respectively. Seven CIS lesions were detected from among all MG-exposed cases and 1 CIS lesion was found in a non-MG lung cancer case. Six of these occurred with dysplasia and 4 were associated with early invasion. Among 62 autopsy cases with known smoking histories, multivariate analysis revealed a significant correlation between the incidence rate of atypia and MG exposure only in non-lung cancer cases: the incidence rate of atypia was also influenced significantly by smoking and age. Among lung cancer cases, the incidence rate of atypia was significantly higher (p less than 0.01) in cases of squamous cell carcinoma than those of small cell carcinoma. PMID- 2985005 TI - [Tumor-targeted chemotherapy with lipid contrast medium and macro molecular anticancer agents theoretical considerations and clinical outcome]. AB - Theoretical considerations for tumor-selective chemotherapy are described which based on the unique character of the tumor neovasculature. Namely, most solid tumors possess four different unique features: hypervasculature, enhanced permeability even to macromolecules, architectural differences, and lack of the lymphatic recovery system. Lipid or lipid contrast medium and macromolecular anticancer agents using prototype drug smancs can be utilized for cancer selective targeting based on the above four features. Selective targeting with lipid contrast medium with smancs has offered two clinical benefits; definite and pronounced antitumor effect and diagnostic value. These effects can be primarily attributed to the tumor-selective accumulation of the agent, i.e., more than 1,000 times greater in the tumor than in the plasma. As a consequence very few side effects are observed clinically. Primary or secondary hepatoma and lung cancer showed size reduction in more than 90 % of treated patients. Very few side effects such as hematosuppression or inhibited liver function were observed in these cases. Prolongation of life-span was marked in the patients. The above results indicate a new future direction for the development of the tumor selective chemotherapy. PMID- 2985006 TI - [Phase I clinical study of NK 171 (etoposide)]. AB - A phase I clinical study of NK 171 (Etoposide), a semi-synthetic derivative of podophyllotoxin, administered by intravenous (i.v.) and oral (p.o.) routes was carried out in patients with malignant tumour who were resistant to standard therapy. As a starting dose of NK 171, 33mg/m2 (expressed as 1n) was administered for 5 consecutive days and the daily dose was increased step by step to a dose of 3.5n i.v. or 5n p.o. respectively. The dose-limiting factor of NK 171 given by the i.v. route was leukopenia and the nadir of leukopenia was observed about 2 weeks after the final day of drug administration in the case of both i.v. and p.o. routes. Recovery from leukopenia required about 3 and 2 weeks from the nadir in the case of i.v. and p.o. administration, respectively. Thrombocytopenia also occurred, but to a lower degree than that observed for leukopenia. A further side effect, alopecia, was observed in about half of the patients. From these results, it was concluded that the recommendable dose schedules of NK 171 for phase II clinical studies are 80-100mg/m2, i.v. and 110-130mg/m2, p.o., respectively, daily for 5 consecutive days and repeated every 4-5 weeks. PMID- 2985007 TI - [Phase II study of recombinant leukocyte A interferon (Ro 22-8181) in skin malignant tumors]. AB - A clinical phase II study of recombinant human leukocyte interferon A (rIFN-alpha A, Ro 22-8181) for various skin malignant tumors was jointly conducted at nine medical institutes across the country in order to study its clinical effect and side effects. Patients received Ro 22-8181 alone in doses ranging from 3 X 10(6) U/day to 50 X 10(6) U/day either by intramuscular injection or by local injection. Good response was obtained in one (4.8%) of 21 patients treated by intramuscular injection and in 26 (72.2%) of 36 patients treated by local injection. The percentage of good responses achieved by local injection for individual diseases was 55.6% (5/9) for metastatic malignant skin melanoma, 100% (11/11) for cutaneous malignant lymphoma, 100% (5/5) for extramammary Paget's disease, 75% (3/4) for intraepidermal cancer and 50% (2/4) for metastatic skin cancer. Main side effects were fever, anorexia, general fatigue, chills, nausea and vomiting. Abnormal laboratory data included leukopenia, and elevation of GOT and GPT, although their incidence was lower with local injection than with intramuscular injection. Side effects were mostly improved by reduction of the dose or discontinuation of the treatment. PMID- 2985008 TI - Normal peripheral-blood lymphocyte cAMP and cGMP resting levels in untreated patients with mycosis fungoides. PMID- 2985009 TI - Glucocorticoid-induced alteration of beta-adrenergic adenylate cyclase response of epidermis. AB - It has been reported that the beta-adrenergic adenylate cyclase system of the pig epidermis is regulated by glucocorticoids, resulting in the augmentation of epinephrine-induced cyclic-AMP accumulations. Using this phenomenon, we compared the glucocorticoidal potency of three typical glucocorticoids: hydrocortisone, prednisolone and dexamethasone. There was a considerable variation in the magnitude of the glucocorticoid-induced augmentation of the beta-adrenergic response when pig skin that had been obtained on different occasions was used. In each experimental series (using the same pig skin), however, the maximal augmentation effects obtained with these glucocorticoids were approximately the same. The potent glucocorticoid, dexamethasone, demonstrated its effect at lower concentrations than were required for prednisolone, while hydrocortisone required a much higher concentration before its effect was detectable. Thus, despite considerable variations in the magnitude of the glucocorticoid effects, the concentrations required for the glucocorticoid effect were closely associated with the established glucocorticoidal potency which has previously been described. PMID- 2985011 TI - Serum angiotensin-converting enzyme activity. Its use in the evaluation and management of hypercalcemia associated with sarcoidosis. AB - Serum angiotensin-converting enzyme (SACE) was measured in 14 patients (eight women and six men) with sarcoidosis and hypercalcemia. Thirteen patients were treated with prednisone, and 12 achieved normal or nearly normal serum calcium values. Two patients had coexistent hyperparathyroidism. Seven of eight patients with serial SACE measurements exhibited parallel falls in SACE and serum calcium levels. Eleven patients were successfully treated with alternate-day prednisone regimens. The data suggest that serial SACE measurements are useful in the evaluation and management of sarcoidosis with hypercalcemia. In patients with sarcoidosis, the reduction of SACE levels during glucocorticoid treatment may be due to a suppression of granuloma formation. Concomitant falls in serum calcium level suggest an important role of the granuloma or its cellular precursors in vitamin D metabolism. PMID- 2985010 TI - An in vitro model for the binding of polybrominated biphenyls in environmentally contaminated blood. PMID- 2985012 TI - [Severe combined immune deficiency with hypereosinophilia. Immunologic study of 5 cases]. AB - We herein report five new cases of severe combined immunodeficiency with hypereosinophilia, the so-called familial reticuloendotheliosis first described by Omenn. It is characterized by erythroderma, polyadenopathy, hepatosplenomegaly, severe and repeated infections, protracted diarrhoea with failure to thrive. There is marked eosinophilia as well as a profound immunodeficiency. The immunologic abnormalities consist of an increase in T cell number, a B cell lymphopenia and a complete lack of humoral and cellular immune responses to antigens. A deficiency of lymphocytes 5'-nucleotidase has been inconstantly found. Histologic findings are characteristic, consisting of severe T and B lymphocyte depletion in lymphoid organs with infiltration by histiocytes and, to a lesser extent, eosinophils. The outcome was uniformly fatal within the first year of life. Treatment by a combination of parenteral nutrition, steroids and epipodophyllotoxin was effective in obtaining the complete remission of clinical manifestations due to the histiocytic and eosinophilic infiltration in two patients. However, the treatment failed to correct the immunologic defect. These results indicate that the histiocytic infiltration is possibly not responsible for the immunologic detect observed in this condition. PMID- 2985013 TI - [Congenital cytomegalovirus infection in dizygotic twins]. PMID- 2985014 TI - Noradrenergic function and the mechanism of action of antianxiety treatment. II. The effect of long-term imipramine treatment. AB - Considerable preclinical and clinical evidence indicates that increased noradrenergic function is involved in the development of anxiety. Imipramine hydrochloride, which has complex effects on noradrenergic function in animals, is effective in patients with agoraphobia and panic disorder. To assess the effects of imipramine on noradrenergic function in patients, plasma levels of free 3 methoxy-4-hydroxyphenylglycol (MHPG) and yohimbine-induced increases in plasma MHPG levels, anxiety-nervousness, blood pressure, and somatic symptoms were studied before and during long-term imipramine treatment in 11 patients meeting DSM-III criteria for agoraphobia with panic attacks. Long-term imipramine treatment significantly decreased baseline plasma MHPG levels by 38% and modestly potentiated yohimbine-induced increases in blood pressure, but it did not alter yohimbine-induced increases in plasma MHPG levels or in patient ratings of anxiety-nervousness. The therapeutic effects of imipramine in panic disorder may relate more to the decrease in norepinephrine turnover than to alterations of alpha 2-adrenergic autoreceptor function. PMID- 2985015 TI - Tritiated imipramine binding distinguishes among subtypes of depression. AB - We studied 45 depressed patients and 20 healthy controls in order to determine if tritiated imipramine binding distinguished among subtypes of primary major depressive disorder. Mean (+/- standard deviation) values for maximal concentration of tritiated imipramine binding sites on platelet membranes were significantly lower in patients with bipolar and familial pure depressive disease (754 +/- 149 and 870 +/- 241 femtomoles [fmole]/mg of protein, respectively) than in patients with depressive spectrum and sporadic depressive disease (1,236 +/- 241 and 1,188 +/- 325 fmole/mg of protein, respectively), neither of which differed from healthy controls (1,238 +/- 201 fmole/mg of protein). Multiple linear regression analysis revealed that these differences could not be attributed to differences in age, sex, Hamilton Rating Scale score, presence of psychotic features, hospitalization status, or medication history. This association of a biological finding with distinct clinically defined subtypes of depression may lead to a classification of affective disorders useful in further research. PMID- 2985016 TI - Research funds are down--take heart! PMID- 2985017 TI - [Antineoplastic chemotherapy for bronchial carcinoma]. AB - With the non small-cell bronchial carcinoma, the results of tumor chemotherapy are, on the whole, still unsatisfactory. It remains to be seen whether the more recent preparations, such as Cisplatin, VP16, Vindesine or combinations there of which are at present extensively investigated, will change this situation. The combinations: Cyclophosphamide/Doxorubicin/Methotrexate/Procarbazine, Cyclophosphamide/Doxorubicin/Cisplatin or Vindesine/Cisplatin are considered most prospective, were remission rates are reported to range from 30-40% with a mean period of remission from 5-7 months. Such values are approximatively reached also by monochemotherapy, e.g. Cyclophosphamide, Adriamycin or Vinblastine. Thus, radiotherapy combined, if necessary, with tumor chemotherapy continues to be the therapy of choice for the non small-cell bronchial carcinoma. Entirely different is the situation with the small-cell bronchial carcinoma. Here, with polychemotherapy being clearly superior to monochemotherapy, 80% of objective remissions are achieved with limited extension of the tumor, and 65%, with advanced tumor progression. The remissions may last here up to 18 months. Combinations of proven value are: Doxorubicin/Cyclophosphamide/Vincristine and VP16/Doxorubicin/Cyclophosphamide, with Cyclophosphamide being contained virtually in all proposed combinations as an essential component. Repeatedly proposed has also been the alternating application of non cross-resistant therapeutic schedules, e.g. VP16/Procarbazine and Cyclophosphamide/Lomustine/Vincristine/Procarbazine. For the inoperable small cell bronchial carcinoma, chemotherapy is the therapy of choice. In how far additional radiotherapy will further improve the therapeutic results is now being intensively investigated. A high likelihood in this regard exists for a "prophylactic" irradiation of the CNS. PMID- 2985018 TI - Diagnostic pathology in the acquired immunodeficiency syndrome. Surgical pathology and cytology experience with 67 patients. AB - We report the pathologic findings in specimens submitted for histologic and cytologic evaluation from 67 patients with the acquired immunodeficiency syndrome. A wide variety of opportunistic pathogens were identified in 41 patients. Mycobacterium avium-intracellulare evoked only a mild host response: granulomas, if present, were poorly formed. Biopsy specimens showing cytomegalovirus gastroenteritis required sections at multiple levels to demonstrate inclusions. Combined histologic and cytologic evaluation can increase the diagnostic yield in pulmonary and esophageal infections. Kaposi's sarcoma was found in biopsy specimens from 29 patients. Early lesions were often extremely subtle, yet distinct from, benign vascular proliferations in involuted lymph nodes. Malignant lymphoma was diagnosed in ten homosexual men who were suspected of having the acquired immunodeficiency syndrome. The lymphomas were characterized by B-cell origin, a diffuse pattern, frequent extranodal presentations, and an aggressive clinical course with prominent central nervous system involvement. PMID- 2985019 TI - Fanconi's anemia, medulloblastoma, Wilms' tumor, horseshoe kidney, and gonadal dysgenesis. AB - An 18-month-old female infant with clinically and cytogenetically documented Fanconi's anemia was found to have two neoplasms previously unreported in this syndrome to our knowledge: a medulloblastoma and a Wilms' tumor, with the latter arising in a horseshoe kidney. An additional feature was pure gonadal dysgenesis. These unusual associations are discussed in the context of certain syndromes suggestive of an axial predisposition for neoplasia (kidneys-central nervous system and kidneys-gonads). PMID- 2985020 TI - Wilms' tumor of the endocervix. AB - We report a case of primary Wilms' tumor of the endocervix in a 13-year-old girl. The tumor was polypoid, filled the vagina, and was attached to the endocervix by a stalk. Microscopic examination disclosed blastematous, epithelial, and stromal elements characteristic of Wilms' tumor. The patient's kidneys were normal on intravenous pyelographic examination and on palpation at laparotomy, and she has remained free of disease for 9.6 years after hysterectomy. PMID- 2985021 TI - Benign glandular inclusions. PMID- 2985023 TI - [Thiophene as a structural element of physiologically active substances. 11. Substituted thieno[2,3-d]imidazoles as potential antiviral agents]. PMID- 2985022 TI - Lung carcinoma by histologic type in coal miners. AB - Histologic types of lung carcinoma were studied in 171 coal miners in the National Coal Workers' Autopsy Study. These miners had an average underground mining tenure of 29 +/- 14 years and an average smoking history of 31 +/- 23 pack years. The proportion of carcinomas by cell type were: squamous cell carcinoma, 30%; adenocarcinoma, 27%; small-cell undifferentiated carcinoma, 26%; large-cell undifferentiated carcinoma, 9%; and other carcinomas, 8%. More tumors were observed in the right lung and in the upper lobes of both lungs than in the left lung and in the lower lobes of both lungs, respectively. The majority of the tumors were centered on cartilaginous airways (81%) as compared with the peripheral regions of the lung (19%). Squamous cell carcinomas predominated in the older miners and in larger airways. Adenocarcinomas were more common in the peripheral lung. No significant interaction was demonstrated between cell type and years of underground mining. The data indicate that lung carcinoma in coal miners differs little in its pathologic features from men in the general population who smoke cigarettes. No effect of coal mine dust exposure on lung carcinoma histogenesis was demonstrated. PMID- 2985024 TI - [Thiophene as a structural element of physiologically active substances. 12. Thiophene analogs of antiviral chalcones]. PMID- 2985025 TI - [Thiophene as a structural element of physiologically active substances. 13. Thiophene analogs of antiviral flavanes]. PMID- 2985026 TI - Studies on potential antiviral compounds, XXIII. 2-(substituted benzoylamino)-3,5 dichloropyridines and isosteric benzamides. PMID- 2985027 TI - Relation of free silicone to human breast carcinoma. AB - Of 12 women with carcinoma of the breast and coexistent silicone mastopathy, nine had had injections of liquid silicone for breast augmentation; three had leaking silicone-gel prostheses. The clinical findings indicated that early diagnosis was obscured by the silicone-induced mastopathy, which rendered the interpretation of physical findings and mammograms difficult. The pathologic findings were suggestive of a possible adverse effect of the presence of free silicone within the breast tissue, axillary nodes, and axillary fat. Although no causal relationship between silicone and breast carcinoma is implied, a heightened awareness of the possible coexistence of silicone mastopathy and breast carcinoma is necessary. PMID- 2985028 TI - Membrane-bound virions of coxsackievirus B4: cellular localization, analysis of the genomic RNA, genome-linked protein, and effect on host macromolecular synthesis. AB - Hela cells infected with several group B coxsackieviruses contain, in addition to standard virions, a population of virus-specific ribonucleoprotein particles which we (5) designated membrane-bound virions (MBV). MBVs differ from standard virions in buoyant density, yield, appearance, protein composition and infectivity. Here we present several new features of MBVs of coxsackievirus B4. The MBVs are lighter (rho about 1.30) and are localized in rough membranes, intermixed with virions. They contain 35S virion RNA covalently linked with a small protein, VPg. The VPg contain two proteins of different charge. MBV VPg is considerably smaller than the 5300-dalton virion VPg. MBV RNA is homologous to the base sequence present in B4 virus double-stranded RNA. The T1 oligonucleotide fingerprint of MBV RNA is distinguishable from that of virion RNA by one oligonucleotide. Several oligonucleotides of virion RNA appear to occur in submolar quantities in MBV RNA. MBVs are 75 to greater than 200 times less infective; they inhibit host cell macromolecular synthesis less efficiently than virions. In coinfected cells, the extent of inhibition of host synthesis is less severe than in cells infected with virions alone, which suggest interference by MBV particles. PMID- 2985029 TI - Isolation from faecal specimens of new strains of human rotavirus primarily cytopathic for stationary cell cultures without trypsin. AB - Eight cytopathic human rotavirus strains were isolated in LLC-MK2 cells and in human embryo fibroblasts. The strains were isolated from faeces collected from pediatric and adult patients. Pretreatment of specimens with trypsin and trypsin incorporation in maintenance medium were not performed. Inoculated monolayers were not subjected to centrifugation and were incubated stationary at 36 degrees C. Viruses were identified by electron microscopy and by fluorescent antibody techniques. It is suggested that these rotaviruses are different from any previously recovered from man. PMID- 2985030 TI - Effect of DNA polymerase inhibitors on the replication of human cytomegalovirus. Brief report. AB - Aphidicolin, a specific inhibitor of cellular DNA polymerase alpha and of viral DNA polymerase, inhibits production of infectious virus and cellular and viral DNA synthesis of human cytomegalovirus (HCMV)-infected cells. On the other hand, 2',3'-dideoxynucleosides, inhibitors of DNA polymerases beta and gamma, do not affect HCMV replication. The data show that the alpha DNA polymerases of either viral or cellular origin are required for viral DNA synthesis, and cannot be substituted by the cellular DNA polymerase beta and gamma. PMID- 2985031 TI - Thermal-pH inactivation of herpes simplex virus: interdependence of the medium composition and the pH on the rate of virus inactivation. Brief report. AB - The composition and pH of the suspending medium were shown to be interdependent effectors of the in vitro thermal inactivation of herpes simplex virus (HSV). In addition, enhanced thermal sensitivity of HSV at alkaline pH was a medium dependent effect and, therefore, is not an inherent property of the virus. PMID- 2985032 TI - Receptor binding characteristics of strains of the influenza Hong Kong subtype, using a periodate sensitivity test. AB - Members of the Hong Kong influenza subtype were analysed for cell-receptor binding activity. This was done by a periodate sensitivity test, involving treatment of fowl erythrocytes with varying concentrations of potassium periodate, before their use in a virus haemagglutination assay. Three classes of receptor binding activity were clearly separated by this test. A 'standard' group comprised early members of the Hong Kong subtype. Two variant groups included later epidemic strains and some lateral terminal strains. Departures from the standard group were correlated with particular amino acid substitutions in the area of the haemagglutinin close to the proposed cell-receptor pocket. In particular, HA1 residue 226 was directly implicated. These amino acid changes were also detectable immunologically. The biological significance of this correlation is discussed. The periodate test is reproducible and could prove to be a useful tool in identifying particular residues in the haemagglutinin molecule which influence binding to the cellular receptor. PMID- 2985033 TI - Immunological and biochemical characterization of human adenoviruses from subgenus B. II. DNA restriction analysis. AB - 47 adenovirus strains related to Ad11, 14, 16, 21, 34, and 35 (including all subgenus B prototypes) were analyzed by DNA restriction analysis with Bam HI, Bgl II, Hind III, and Sma I. Typical fragments occurring in most of the strains were observed only with Hind III. Besides Ad3 and 7, 20 different genome types were distinguished. A pairwise analysis of comigrating fragments was performed for all genome types and summarized for the four enzymes. A medium rate of comigrating fragments (50 to 60 per cent) was found between Ad11, 14, 34, and 35 strains; Ad16 and 21 had 35 to 50 percent comigrating fragments. Within each group of closely related strains, Ad16 strains, Ad21 strains, and Ad34 + 35 strains showed a high rate (greater than 70 per cent) of comigrating fragments. Ad3 and 7 showed more comigration with Ad16 and 21 than with the others. Sixteen strains related to Ad21 were identical in their DNA restriction pattern and by serology. The relatedness determined by fragment comigration and by serology was compared by a quantitative estimate. The DNA relationship between Ad11 + 14 and Ad34 + 35 was not corroborated by serology. Ad21 was serologically related to Ad34 and 35, but not to Ad16; the opposite was found for DNA. The intermediate Ad21-16 showed a unique DNA pattern with some relation to Ad16. PMID- 2985034 TI - Bovine herpesvirus 1: differentiation of IBR- and IPV-viruses and identification and functional role of their major immunogenic components. AB - Infectious Bovine Rhinotracheitis (IBR) and Infectious Pustular Vulvovaginitis (IPV) virus strains of Bovine Herpesvirus 1 (BHV-1) can be differentiated by restriction endonuclease digestion of their DNAs. Antigens and polypeptide patterns of isolates of these different clinical entities are almost identical. Page analysis of immunoprecipitates revealed three major immunogenic components in BHV-1 infected cells. These are glycoproteins with apparent molecular weights of 93,000 (GP93), 74,000 (GP74) and 69,000 daltons (GP69), respectively. Bovine convalescent sera and antisera, which are directed against individual precipitates derived from crossed immunoelectrophoresis, contain antibodies reacting with one or more of these glycoproteins. The experiments with these antisera demonstrate that GP74 and possibly GP93, both structural components of the BHV-1 virion, induce neutralizing antibodies, whereas GP69, a non-structural protein, does not. PMID- 2985035 TI - Chemotherapy for aggressive juvenile nasopharyngeal angiofibroma. AB - Five patients with recurrent juvenile nasopharyngeal angiofibromas were treated with antineoplastic chemotherapy. All patients had tumor remission without recurrence; there were no sequelae after this therapy. PMID- 2985036 TI - Prostaglandins and other metabolites of arachidonic acid. An overview for the otolaryngologist. AB - Metabolites of arachidonic acid have important regulatory functions within several areas of concern to the otolaryngologist. Prostaglandins, composing one group, are involved in smooth-muscle contraction, regulation of renal glomerular blood flow, and in the modulation of immune and allergic responses, inflammation, fever, pain, and tumor growth. A second group, the leukotrienes, may be even more important than prostaglandins in allergy and inflammation. The elusive slow reacting substance of anaphylaxis belongs in this group. Two other metabolites, thromboxane and prostacyclin, seem to be critical in hemostasis and the metastatic spread of tumors. PMID- 2985037 TI - Effects of chemically or virus-induced immunodepression on response of chickens to avian leukosis virus. AB - The effects of chemically or virus-induced immunodepression on the infection profile (development of viremia and antibody) and shedding of avian leukosis virus (ALV) were studied in progeny chickens of experimental or commercial breeder flocks. Chickens were infected with ALV subgroup A by contact at hatching and by oral inoculation at 4-5 weeks of age. In the first experiment, chickens were inoculated with a virulent strain of infectious bursal disease virus (IBDV) at 1 day or 6 weeks of age. In the second experiment, chickens were neonatally treated with cyclophosphamide (CY), or were inoculated with strain T of reticuloendotheliosis virus (REV) at hatching, or were inoculated with strain JM of Marek's disease virus (MDV) at 2 weeks of age. The infection profile and cloacal shedding of ALV in chickens exposed to ALV and inoculated with immunodepressive viruses or CY were compared with those in hatchmates exposed only to ALV. In two of four chicken lines tested in the first experiment, shedding of ALV, as determined by virological assays of cloacal swabs at 22 weeks of age, was significantly higher in chickens infected with IBDV at 1 day of age than in uninfected hatchmates. The rate of shedding of ALV in one of these two lines was also significantly higher in chickens infected with IBDV at 6 weeks of age than in uninfected chickens. Further, the frequency of ALV-antibody detection at 22 weeks of age was significantly lower in chickens of these two lines infected with IBDV at 1 day of age than in uninfected chickens. In the second experiment, neonatal treatment with CY significantly increased the frequency of viremic chickens of both experimental and commercial flocks. The frequency of ALV viremic chickens at 22 weeks of age was considerably higher in the REV- and MDV inoculated groups (54% and 44%, respectively) than in control hatchmates (29%), but only in chickens of the commercial line. These findings suggest that chemically or virus-induced immunodepression may lead to an increase in rates of viremia and shedding of ALV in chickens infected with virus after hatching, especially in certain genetic lines. PMID- 2985038 TI - Adenovirus-like particles associated with intranuclear inclusion bodies in the kidney of a common murre (uria aalge). AB - Adenovirus-like particles were identified by transmission electron microscopy in intranuclear inclusion bodies in the renal collecting tubules of a male common murre. The bird had been trapped in an oil-spill and had been cleaned and held in captivity before euthanasia. The presence of the virus appeared to be causing little or no renal disease. It is thought that this bird suffered activation of a latent viral infection as a result of the stress of oil intoxication, handling, and confinement. PMID- 2985039 TI - Respiratory disease (rhinotracheitis) of turkeys in Brittany, France. III. Interaction of multiple infecting agents. AB - A candidate breeder flock of turkeys was studied during and after an outbreak of rhinotracheitis. Laboratory studies revealed the presence of three pathogens during the acute phase of the disease. These agents were hemorrhagic enteritis virus (HEV), paramyxovirus type 2 (PMV2), and chlamydia. Twenty-five turkeys in the flock were individually identified, and blood samples from these birds were collected for serological studies each week for 21 weeks. The serological results revealed high titers for HEV and chlamydia but very low titers for PMV2. PMID- 2985040 TI - Enzyme immunoassay studies on the serological response of turkeys to hemorrhagic enteritis virus. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of antibodies in turkey serum to hemorrhagic enteritis virus. The ELISA antigen was extracted from turkey spleens and partially purified with fluorocarbon. Antibodies were demonstrated in serum samples of breeding and meat flocks that had been naturally exposed to infection. These samples were also examined in parallel by agar-gel precipitin (AGP); most of the sera were AGP-positive. ELISA, however, was more sensitive in detecting antibodies in day-old sera that were AGP negative. The passively acquired antibodies were no longer detected by 4 weeks of age. A brisk but short-lived secondary response was detected by ELISA in the sera of turkeys immunized with beta-propiolactone-inactivated extract of infected spleens. PMID- 2985041 TI - The influence of endurance training on mechanical catecholamine responsiveness, beta-adrenoceptor density and myosin isoenzyme pattern of rat ventricular myocardium. AB - An investigation was carried out on the effects of 4 weeks' swimming training (2 X 90 min/day) on myocardial isometric tension development and rate of tension rise, and also on the changes induced therein by in vitro application of isoproterenol. This was done in 9 isolated papillary muscles of 9-week-old male Wistar rats and the results were compared with the data of age-matched sedentary controls. Ventricular beta-adrenoceptors [( 3H]-dihydroalprenolol binding) and the isoenzyme pattern of myosin (pyrophosphate gel electrophoresis) were examined in the same individuals. Isometric tension (T) and its first derivative (dT/dt) measured at the optimum of the length-tension diagram were moderately increased by long-term swimming training. Isoproterenol (10(-5) mol/l) induced a greater absolute and relative increase of both mechanical parameters in specimens of trained animals than in age-matched controls (delta T: 3.6 +/- 1.6 vs. 1.9 +/- 0.6 X 10(-2) N/mm2, p less than 0.05. delta dT/dt: 43.4 +/- 14.0 vs. 30.4 +/- 9.5 X 10(-2) N/mm2 X s, p less than 0.05). KD decreased significantly (4.23 +/- 1.0 vs. 2.44 +/- 0.3 nM, p less than 0.02), indicating an increase in receptor affinity, whereas receptor density revealed a tendency to decrease (98.8 +/- 22.6 vs. 67.1 +/- 18.0 fmol/mg protein, p less than 0.1). In addition, there was a shift in the isoenzyme pattern of myosin towards VM-1 after swimming training. Thus, under the conditions of the present experiments, the mechanical response to isoproterenol does not correlate to beta-adrenoceptor density. It is probable that, apart from the altered sensitivity of the receptors, other membrane or post membrane processes, are responsible for the increased mechanical responsiveness to catecholamines. Although a relationship between myosin isoenzyme pattern and mechanical responsiveness to catecholamines is apparent taking into account our results and the findings on hypertensive rats as reported in the literature, it cannot be accounted for simply by altered beta-adrenoceptor density. PMID- 2985042 TI - Low apparent aldose reductase activity produced by monosaccharide autoxidation. AB - Low apparent aldose reductase activity, as measured by NADPH oxidation, can be produced by the spontaneous autoxidation of monosaccharides. NADPH is oxidized to metabolically active NADP+ in a solution of autoxidizing DL-glyceraldehyde at rates of up to 15 X 10(-4) A340/min. The close parallelism between the effects of buffer salt type and concentration, monosaccharide structure and temperature activation on autoxidation and NADPH oxidation imply that autoxidation is a prerequisite for the NADPH oxidation, probably via the hydroperoxy radical. Nucleotide-binding proteins enhanced NADPH oxidation induced by DL glyceraldehyde, up to 10.6-fold with glucose-6-phosphate dehydrogenase. Glutathione reductase-catalysed NADPH oxidation in the presence of autoxidizing monosaccharide showed many characteristics of the aldose reductase reaction. Aldose reductase inhibitors acted as antioxidants in inhibiting this NADPH oxidation. These results indicate that low apparent aldose reductase activities may be due to artifacts of monosaccharide autoxidation, and could provide an explanation for the non-linear steady-state kinetics observed with DL glyceraldehyde and aldose reductase. PMID- 2985043 TI - Characterization and metabolic regulation of a liver-specific 5.4-kilobase mRNA whose synthesis is transcriptionally induced by carbohydrates and repressed by glucagon and cyclic AMP. AB - Four clones derived from a carbohydrate-induced rat liver cDNA library were found to hybridize with a 5.4-kilobase mRNA species encoding a 36 kDa protein. This mRNA was abundant in the liver, barely detectable in adipocytes and kidney, and absent from the other tissues tested. In the liver, the mRNA was fully induced by a carbohydrate-rich diet, but was undetectable during both starvation and feeding with a protein-rich or lipid-rich diet. Adrenalectomized, thyroidectomized and diabetic animals did not express the mRNA in their liver when re-fed with the carbohydrate-rich diet. When these animals were given the missing hormone, the amount of hybridizable RNA returned to normal values, but administration of the hormone alone failed to induce mRNA synthesis in starved animals. Both glucagon and its second messenger, cyclic AMP, abolished the induction of the mRNA in re fed animals. Exogenous insulin, whatever the dose, did not reverse the inhibitory action of glucagon. In an isolated nuclei transcription system, no detectable RNA transcripts were found in starved animals, whereas feeding the animals with the carbohydrate-rich diet led to a maximum rate of gene transcription. Although unidentified, this mRNA proves to be a remarkable marker of dietary and hormonal control of gene expression in vivo. It will provide a useful model for further analysis of the role of cyclic AMP in regulating the transcription of eukaryotic genes. PMID- 2985044 TI - 5'-Nucleotidase from rat heart membranes. Inhibition by adenine nucleotides and related compounds. AB - ADP and ATP and their analogues were evaluated as inhibitors of 5'-nucleotidase purified from heart plasma membrane. ADP analogues are more powerful inhibitors than the corresponding ATP analogues. The most powerful inhibitor found is adenosine 5'-[alpha beta-methylene]diphosphate (AOPCP) for which the enzyme shows a Ki of 5 nM at pH 7.2. Measurements of pKi values for ADP and AOPCP as a function of pH indicate that the major inhibitory species of both nucleotides is the dianion. In the physiological range of pH values, AOPCP is a more powerful inhibitor than ADP principally because a higher percentage of AOPCP exists in the dianion form. The methylenephosphonate analogue of AMP (ACP), though not a substrate, is a moderately effective inhibitor. The corresponding analogues of ADP (ACPOP) and ATP (ACPOPOP) are as good inhibitors as ADP and ATP respectively. The thiophosphate analogues of ADP all inhibit 5'-nucleotidase, although not as powerfully as ADP, the most effective of these analogues being adenosine 5'-O-(1 thiodiphosphate) diastereoisomer B (ADP[alpha S](B)]. Other nucleotides inhibit the enzyme, but none is as effective as AOPCP. Inorganic tripolyphosphate and methylenediphosphonate are better inhibitors of the enzyme than is inorganic pyrophosphate. Inorganic thiophosphate is a better inhibitor than is orthophosphate. Hill plots of the ADP and AOPCP inhibition yield slopes close to 1; Hill plots of the ATP inhibition yield slopes of about 0.6. MgADP- is not an inhibitor, and MgATP2- is at best a very weak inhibitor of the enzyme. PMID- 2985045 TI - The effect on photohaemolysis of variation in the structure of the porphyrin photosensitizer. AB - A comparison of the photosensitizing ability of a variety of porphyrins for photohaemolysis gives the following order of activity: protoporphyrin greater than deuteroporphyrin, mesoporphyrin, haematoporphyrin dimethyl ester much greater than haematoporphyrin diacetate, haematoporphyrin greater than haematoporphyrin monoacetate, coproporphyrin III, haematoporphyrin derivative, coproporphyrin III tetramethyl ester greater than uroporphyrin I, meso-tetra-(N methyl-4-pyridinium)porphyrin tetratoluene-p-sulphonate, meso-tetra-(p carboxyphenyl)porphyrin, protoporphyrin dimethyl ester, meso-tetra-(p-hydroxy sulphonylphenyl)porphyrin tetrasodium salt, uroporphyrin III, deuteroporphyrin 3,8-disulphonic acid and protohaemin. The results for the metal-free porphyrins are rationalized in terms of solubility and partition properties, and a model is proposed for the incorporation of amphipathic porphyrins into the membrane lipid bilayer. Experiments with erythrocytes from patients with erythropoeitic protoporphyria and with normal erythrocytes to which porphyrin was added in a deuterium oxide medium do not lead to an increase in the rate of photohaemolysis. A possible explanation for this somewhat surprising observation is outlined. PMID- 2985046 TI - Regulatory proteins (inhibitors or activators) affect estimates of Mr of enzymes and receptors by radiation inactivation. A theoretical model. AB - The radiation-inactivation method allows the determination of the Mr of enzymes and receptors by monitoring the decay of biological activity as a function of absorbed dose. The presence of regulatory or effector proteins (inhibitors or activators) associated with an enzyme or receptor, or released in the preparation after tissue homogenization, may affect the decay of biological activity. How the activity is affected, however, will depend on the type of inhibition (competitive or non-competitive), the inhibitor or activator concentration, the dissociation constant of the enzyme-effector system, and the effector Mr relative to that of the enzyme. Since little is known on how effector proteins influence radiation inactivation of enzymes and receptors, we have considered a theoretical model in an effort to provide a framework for the interpretation of experimentally obtained data. Our model predicts that competitive and non-competitive inhibitors of enzymes could be distinguished by analysing irradiated samples with various substrate concentrations. Inhibitors will decrease whereas activators will increase the apparent target size of enzymes or receptors. PMID- 2985047 TI - N-Oxidation of 4-chloroaniline by prostaglandin synthase. Redox cycling of radical intermediate(s). AB - 4-Chloroaniline undergoes N-oxidation in ram seminal-vesicle microsomal preparations supplemented with arachidonic acid to yield N-(4-chlorophenyl) hydroxylamine and 1-chloro-4-nitrosobenzene. H2O2 also supports metabolism of the amine substrate to the same organic-solvent-extractable products, suggesting that the hydroperoxidase activity of prostaglandin synthase is responsible for the co oxidation. Analysis of the reaction mixtures by e.s.r. spectrometry reveals the formation of a radical intermediate bearing the characteristics of a strongly immobilized nitroxide. Arylamine-stimulated O2.- release can be observed when the arachidonic acid-containing incubation media are supplemented with NADPH. Redox cycling of the nitroxide/hydroxylamine couple is presumed to represent the major source of O2.-, but additional mechanisms, such as redox changes of nitro anion radicals resulting from potential further metabolism of 1-chloro-4 nitrosobenzene, cannot be excluded. The concerted action of carrier-bound nitroxides and O2.- in initiating damage of cellular macromolecules is discussed. PMID- 2985048 TI - Cardiac sarcoplasmic-reticulum calmodulin-binding proteins. Modulation of calmodulin binding to phospholamban by phosphorylation. AB - The gel-overlay technique with 125I-labelled calmodulin allowed the detection of several calmodulin-binding proteins of Mr 280 000, 150 000, 97 000, 56 000, 35 000 and 24 000 in canine cardiac sarcoplasmic reticulum. Only two calmodulin binding proteins could be identified unambiguously. Among them, the 97 000-Mr protein that undergoes phosphorylation in the presence of Ca2+ and calmodulin, is likely to be glycogen phosphorylase. In contrast, the (Ca2+ + Mg2+)-activated ATPase did not appear to bind calmodulin under our experimental conditions. The second known calmodulin target is dephosphophospholamban, which migrates with an apparent Mr of 24 000. The dimeric as well as the monomeric form of phospholamban was found to bind calmodulin. Phospholamban shifts the apparent Kd of erythrocyte (Ca2+ + Mg2+)-activated ATPase for calmodulin, suggesting thus a tight binding of calmodulin to the proteolipid. Interestingly enough, phospholamban phosphorylation by either the catalytic subunit of cyclic AMP-dependent protein kinase or the Ca2+/calmodulin-dependent phospholamban kinase was found to inhibit calmodulin binding. PMID- 2985049 TI - Topoisomerase I is the predominant nuclear protein from avian erythrocytes that can be covalently linked to DNA. AB - Nuclear extracts of erythrocytes contain proteins which stably or possibly covalently bind to DNA. These proteins can be detected by an assay which was originally developed to quantify stable binding of topoisomerases to DNA [Trask, DiDonato & Muller (1984) EMBO J. 3, 671-676]. In this report, we show that the number of activities detected by this assay in crude extracts of nuclei is limited predominantly to various forms of topoisomerase I. One form, a 50 kDa protein, copurifies with histone H1. Western blotting experiments suggest that the 50 kDa topoisomerase exists in chromatin along with the 105 kDa form. In addition, the ratio between the high and low-Mr forms is relatively constant in erythrocytes and embryonic fibroblasts. These results imply that the multiple forms are not unique to one tissue setting. PMID- 2985050 TI - Mechanism of the superoxide-producing oxidase of neutrophils. O2 is necessary for the fast reduction of cytochrome b-245 by NADPH. AB - A soluble oxidase from phorbol-stimulated pig neutrophils contained FAD and cytochrome b-245. A typical preparation produced 13.03 mol of superoxide (O2-.) X S-1 X mol of cytochrome b-1 (348 nmol X min-1 X mg of protein-1). In the aerobic steady state, cytochrome b was 8.9% reduced. Steady-state cytochrome b reduction was absent from extracts of unstimulated cells; Km values for NADPH, for O2-. production and cytochrome b reduction were similar. The calculated aerobic rate of cytochrome b reduction was equal to the measured rate of O2-. production in a variety of preparations and in the presence of a range of inhibitors. Under anaerobic conditions the rate was slow: O2 is apparently required for rapid electron flow into the oxidase complex. Cytochrome b is shown to be kinetically competent to act as part of the O2-.-generating complex. PMID- 2985051 TI - Archaebacterial malate dehydrogenases. The enzymes from the thermoacidophilic organisms Sulfolobus acidocaldarius and Thermoplasma acidophilum show A-side stereospecificity for NAD+. AB - The stereoselective transfer of hydrogen from NADH to oxaloacetate catalysed by malate dehydrogenases (EC 1.1.1.37) from the thermoacidophilic archaebacteria Sulfolobus acidocaldarius and Thermoplasma acidophilum was studied by the p.m.r. method described by Zhou & Wong [(1981) J. Biochem. Biophys. Methods 4, 329-338]. Both enzymes are A-side (pro-R) stereospecific for NADH. PMID- 2985052 TI - The specificity of yeast low-Km cyclic AMP phosphodiesterase towards free bivalent metal ions and the diastereoisomers of cyclic adenosine phosphorothioate. AB - The relative activity of a zinc-containing cyclic AMP phosphodiesterase towards the (Sp)- compared with the (Rp)-diastereoisomer of cyclic adensine phosphorothioate varied with the identity of the free bivalent metal ion from more than 35 to 0.074 along the series Mg2+ greater than Mn2+ greater than Co2+ greater than Zn2+ greater than Cd2+, showing that this ion, and not the tightly bound zinc, bonds to the phosphorothioate moiety of the substrate. PMID- 2985053 TI - IHF stimulation of lambda cII gene expression is inhibited by the E. coli NusA protein. AB - The effects of E. coli proteins Integrative Host Factor (IHF) and NusA on the regulation of lambda cII gene expression are presented. As reported previously (Peacock et al. [1984] Proc. Natl. Acad. Sci. USA 81, 6009-6013), IHF stimulates the DNA-directed in vitro synthesis of cII protein or its first dipeptide, fMet Val. Whereas NusA, by itself, has no effect on cII expression, the presence of NusA inhibits the IHF-mediated stimulation of cII synthesis. PMID- 2985054 TI - Inhibition of aldosterone production by alpha-human atrial natriuretic polypeptide is associated with an increase in cGMP production. AB - Synthetic alpha-human atrial natriuretic polypeptide caused rapid and marked inhibition of aldosterone production in dispersed rat adrenal capsular cells. The polypeptide also slightly, but significantly, decreased cAMP production in the adrenal dispersed capsular cells, while markedly stimulating cGMP production. The cGMP production was accelerated at the concentration of alpha-human atrial natriuretic polypeptide lower than the threshold level to stimulate aldosterone production. These findings suggest that alpha-human atrial natriuretic polypeptide possibly plays a regulatory role in aldosterone production and an additional role in natriuresis through inhibition of aldosterone production. The stimulation of cGMP production by alpha-human atrial natriuretic polypeptide may be involved in the inhibitory effect of this peptide on aldosterone production. PMID- 2985055 TI - Cystatin, a protein inhibitor of cysteine proteases alters viral protein cleavages in infected human cells. AB - Chicken cystatin is a previously described small protein which has the property of inhibiting cysteine active site proteases. The protein, when added to cultured human cells, alters the intracellular proteolytic processing of poliovirus proteins, and causes a reduction in virus yield. It is suggested that the cystatin is able to penetrate to the cellular cytoplasm and inhibit the action of the poliovirus protease. PMID- 2985056 TI - Na+/H+ exchanger activity in the pig kidney epithelial cell line, LLC-PK1: inhibition by amiloride and its derivatives. AB - Rapidly growing pig-kidney-derived epithelial cells, LLC-PK1, lack detectable amiloride-sensitive Na+/H+ exchange activity when assayed directly. A large 22Na uptake is induced when the cells are acid-loaded prior to assay by incubation with buffer containing ammonium chloride or nigericin. The acid-stimulated sodium uptake is sensitive to amiloride, with half-maximal inhibition at 3.5-4.5 microM in buffer containing 15 mM sodium ion. There is simple competitive interaction between amiloride and sodium ion when the amiloride concentration is below 25 microM and the sodium ion concentration is above 20 mM. Derivatives of amiloride which carry substituents on the 5-amino group are 35- to 175-fold more inhibitory than amiloride itself. PMID- 2985057 TI - Spin-labeled erythrocyte membranes: direct identification of nitroxide-conjugated proteins. AB - The covalent incorporation of a spin-labeled analog of N-ethyl maleimide into erythrocyte membrane proteins has been monitored by electron paramagnetic resonance spectroscopy and the individually labeled proteins detected by immunoblotting techniques, using an anti-nitroxide antibody, following electrophoretic separation of the membrane components. Spin-label was primarily found in the high molecular weight bands (I and II) with no incorporation in proteins with molecular weights less than 35,000. Increasing the reaction time between the spin-label and ghosts altered both the observed labeling pattern and the epr spectra with an increase in the proportion of strongly-immobilized species. PMID- 2985058 TI - Platelet activating factor (PAF) involvement in endotoxin-induced hypotension in rats. Studies with PAF-receptor antagonist kadsurenone. AB - Evidence from three types of experiments indicates that platelet activating factor (PAF)1 is an important mediator of endotoxin-induced hypotension in rats. a) Endotoxin infusion stimulates the time-dependent appearance of PAF in the blood. b) PAF infusion results immediately (less than 30 sec) in hypotension while endotoxin-induced hypotension takes 3-5 min to occur, allowing time for PAF production. c) Infusion of the specific PAF-receptor antagonist kadsurenone (2.2 mumole/kg bolus, 0.9 mumoles/min/kg continuous infusion), which inhibits PAF induced hypotension by 67%, causes a 67% reversal of endotoxin-elicited hypotension. An additional finding of this study is that rats respond hypotensively to each of a series of low-dose PAF infusions but only to the first low-dose endotoxin infusion. These endotoxin-refractory rats do respond to subsequent PAF infusions. PMID- 2985059 TI - Human leukocyte interferon inhibits human chorionic gonadotropin stimulated testosterone production by porcine Leydig cells in culture. AB - Pretreatment of primary porcine Leydig cell cultures with human leukocyte interferon suppressed the subsequent hCG-stimulated testosterone production in a dose-dependent manner, with an ED50 at 13 IU/ml. The treatment had no effect on hCG-binding to its receptor, and the inhibition of testosterone production was not abolished by 8Br-cAMP addition. The results indicate that the site of interferon action on hCG-stimulated testosterone production in primary cultures of porcine Leydig cells is located distal to cAMP formation. PMID- 2985060 TI - Light-induced conformational changes in the extradiscal regions of bovine rhodopsin. AB - Light-induced conformational changes occurring at the cytosolic surface of rhodopsin were investigated by performing limited digestions of native and illuminated visual pigment with thermolysin, Arg-C endoproteinase, papain and proteinase K. A higher susceptibility of the extradiscal regions of the bleached pigment to the proteases were observed together with altered capacities of the digested bleached rhodopsins to activate the cGMP phosphodiesterase. The overall results strongly suggest that light induces conformational changes not only in the C-terminal end but also in the second and the third extradiscal loop of rhodopsin. PMID- 2985061 TI - Solubilisation of ferricytochrome c in methanol using a crown ether: absorption, circular dichroism and EPR spectral properties. AB - Ferricytochrome c is normally insoluble in methanol, but its solution is facilitated by complexation with 18-crown-6. Absorption, circular dichroism and EPR spectroscopy indicate that the solubilised protein in MeOH exists in at least three conformational states, all different from the native state in neutral aqueous solution. In two states the haem iron (III) is low spin and in one state it is high spin, but it seems likely that all three forms are globular. The proportion in the high spin form increases at increasing crown ether concentration and on ageing the protein solution. The protein appears to return to its native conformation when it is restored to an aqueous environment. PMID- 2985062 TI - Occlusion of Na+ by the Na,K-ATPase in the presence of oligomycin. AB - Oligomycin occludes Na+ in an E1-form of the Na,K-ATPase. The rate constants for the release of Na+ from the E1-form and for the transition to the E2-form are about 0.5 s-1. The effect of oligomycin is not seen using other cations which also have a Na+-like effect on the enzyme conformation. The inhibitory effect of oligomycin on the ADP-ATP dependent Na:Na exchange but not on the accompanying ADP-ATP exchange can be explained from a decrease in the rate of release of Na+ from an E1 approximately phosphoform with Na+ occluded, E'1 approximately P (Na3), i.e. with Na+ in the membrane phase, to an E"1 approximately PNa3 form with Na+ not occluded. E"1 approximately PNa3 is at a step before formation of E2 P, and disappears at a high rate when ADP reacts with E"1 approximately P (Na3). PMID- 2985063 TI - Comparison of ADP-ribosylation of chromosomal proteins between intact and broken cells. AB - ADP-ribosylation of nonhistone high mobility group (HMG) proteins and histone H1 in intact cells was markedly different from that in the broken cell systems using isolated nuclei and chromatin. (i) The amounts of (ADP-ribose)n on these proteins in intact cells were less 10(-2) - 10(-3) of those in the broken cell systems. (ii) The modified protein molecules in intact cells were about 2 orders of magnitude less extent than those in the broken cells. (iii) The (ADP-ribose)n chains synthesized in intact cells, nuclei and chromatin were mainly mono, oligo and poly, respectively. (iv) The principal acceptor molecules in intact cells were concentrated in HMG 1, 2, 14 and 17, and histone H1, whereas the four HMG proteins were minor acceptors in nuclei, and HMG 1 and 2 and histone H1 were major acceptors in chromatin. PMID- 2985064 TI - Inhibition of renin secretion by platelet activating factor (acetylglyceryl ether phosphorylcholine) in cultured rat renal juxtaglomerular cells. AB - Acetylglyceryl ether phosphorylcholine (AGEPC), commonly known as platelet activating factor, was found to strongly inhibit renin secretion in cultures rich in juxtaglomerular cells. This inhibitory action of AGEPC was accompanied by an enhanced calcium permeability of the cell membrane as evaluated from measurements of the uptake of 45Ca. Simultaneous addition of the calcium channel blocker verapamil abolished the effects of AGEPC on both renin secretion and calcium permeability. Furthermore, addition of AGEPC to the cell cultures led to a decrease of 32P-labeled phosphatidylinositol 4,5-bisphosphate and to an increase in 3H-labeled diacylglycerol, indicating an activation of phospholipase C by AGEPC. PMID- 2985065 TI - Interactions of arachidonate metabolism and protein kinase C in mediating neutrophil function. AB - Diglyceride activators of protein kinase C (i.e., 1-0-myristoyl-, 1-0-palmitoyl-, and 1-0-oleoyl-2-acetylglycerol) interacted synergistically with an arachidonate metabolite, 5-hydroxyicosatetraenoate, to stimulate neutrophil degranulation and superoxide anion generation. Contrastingly, combinations of 15 hydroxyicosatetraenoate with the glycerides or 5-hydroxyicosatetraenoate with a dialkylglyceride (1-0-hexadecyl-2-ethylglycerol) produced no such synergy. The data support a view of stimulus-response coupling wherein protein kinase C is activated in parallel with the mobilization of arachidonate. Respective products of these events, e.g., phosphorylated proteins and hydroxyicosatetraenates, then interact to mediate function. PMID- 2985066 TI - Lack of inhibition by atrial natriuretic factor on cyclic AMP levels in single nephron segments and the glomerulus. AB - Recently an inhibitory effect of atrial natriuretic factor (ANF) on the adenylate cyclase system has been reported in vascular tissue. In seeking similar affects in renal tissue, we studied the effect of ANF on cyclic AMP levels in single nephron segments and in glomeruli from the rat. Individual nephron segments or glomeruli were incubated in the presence of a phosphodiesterase inhibitor, with or without parathyroid hormone (PTH) or arginine vasopressin (AVP) and varying concentrations of ANF at 37 degrees C for 2 min. The capacity for alpha 2 adrenoceptor inhibition of adenylate cyclase was demonstrated in the proximal convoluted tubule, cortical collecting tubule and in glomeruli. Nevertheless, ANF could not inhibit cAMP formation in any of these nephron segments nor in the glomerulus. Thus, unlike the vasculature, ANF has no inhibitory effect on cAMP formation in these renal tissues. PMID- 2985067 TI - Rat hypothalamic GRF elicits its biologic action in GH3 cells by interaction with VIP- preferring receptor site(s). AB - GH3 cells can be used effectively to study the in vitro mechanism of action of GRF. In these cells, there is a time and concentration-dependent release of cAMP into the medium. Rat hypothalamic GRF, (rGRF) is 7 to 10 fold more active than human hypothalamic GRF (hGRF). VIP, a peptide which is structurally homologous to GRF, stimulates cAMP efflux in GH3 cells, with a higher affinity than hGRF or rGRF. We propose that in contradistinction to the normal rat pituitary, the stimulation of cAMP release by GRF in GH3 cells occurs via activation of VIP preferring receptors and that GRF (rGRF in particular) behaves as a partial VIP agonist. PMID- 2985068 TI - Identification of the short dispersed repetitive DNA sequences isolated from the zones of initiation of DNA synthesis in human cells as Alu-elements. AB - DNA of Xeroderma pigmentosum cells was crosslinked in vivo with trioxsalen and long wave length ultraviolet light and the cells were cultured in the presence of labelled thymidine for one hour. The nascent DNA chains synthesized during this period and containing the DNA replication origins were isolated from the high molecular weight chromosomal DNA by an alkaline sucrose density gradient centrifugation. They were 5-10-fold enriched in short dispersed repetitive sequences identified by dot-blot hybridization to BLUR 8 plasmid as members of the human Alu-family. PMID- 2985069 TI - Binding of cytochrome c2 to the isolated reaction center of Rhodospirillum rubrum involves the "backside" of cytochrome c2. AB - Lys 109, Lys 112 and Glu 1 of cytochrome c2 from Rhodospirillum rubrum G-9 are about 4-fold less reactive towards acetic anhydride when cytochrome c2 is bound to the isolated photosynthetic reaction center from the same organism. The three shielded residues are clustered together on the "backside" of cytochrome c2. This contrasts with mitochondrial cytochrome c where "frontside" lysines are protected by different physiological electron transfer partners. PMID- 2985070 TI - Ca2+-dependent stimulation of 3-O-methylglucose transport in mouse fibroblast Swiss 3T3 cells induced by phorbol-12,13-dibutyrate. AB - Binding of phorbol-12,13-dibutyrate (PDBu), a tumor promoter, to quiescent Swiss 3T3 cells increased the number of hexose carriers, resulting in stimulation of membrane transport of 3-O-methylglucose (3MeGlu) in a Ca2+-dependent fashion. Extracellular Ca2+ was necessary to initiate the binding of PDBu to its receptor, and intracellular Ca2+ was required to maintain it. The loss of PDBu-binding, caused by elimination of Ca2+, was accompanied by a loss of stimulation of hexose transport. These results indicated that Ca2+-dependent, continuous binding of PDBu to its receptor was essential to induce the stimulation of hexose transport. PMID- 2985071 TI - Ca2+- or phorbol ester- dependent effect of ATP on a subpopulation of cAMP cell surface receptors in membranes from D. discoideum. A role for protein kinase C. AB - D. discoideum cells contain surface receptors for the chemoattractant cAMP which are composed of fast and slowly dissociating binding sites with half-lifes of respectively about 1 s and 15 s (Van Haastert and De Wit, J. Biol. Chem. 259, 13321-13328). In membranes prepared by shearing the cells through a Nucleopore filter, ATP has no effect on cAMP-binding at equilibrium, but the number of slowly dissociating sites is increased about 2-fold by ATP while their apparent affinity and off-rate are not altered by ATP. The effect of ATP is stimulated about 3-fold by Ca2+ with a half maximal effect at 100 microM Ca2+. The tumor promoting phorbol ester, phorbol 12-myristate 13-acetate (PMA), increases this Ca2+-sensitivity of the ATP effect to about 0.2 microM Ca2+. These data suggest that a specific subpopulation of cAMP receptors in membranes from D. discoideum is altered by the action of protein kinase C. PMID- 2985072 TI - Low temperature magnetic circular dichroism spectroscopy as a probe for the optical transitions of paramagnetic nickel in hydrogenase. AB - A partially-purified sample of hydrogenase from Methanobacterium thermoautotrophicum (delta H strain) has been investigated by optical absorption, magnetic circular dichroism and electron paramagnetic resonance spectroscopy. Variable temperature magnetic circular dichroism studies reveal, for the first time, the optical transitions associated with the Ni(III) center in the oxidized enzyme. Low temperature magnetic circular dichroism spectroscopy provides a new method of assessing both the coordination environment of Ni in hydrogenase and the appropriateness of inorganic model complexes. PMID- 2985073 TI - The specific vulnerability of the substantia nigra to MPTP is related to the presence of transition metals. AB - We demonstrate that the high concentration of transition metals in the substantia nigra could be a major factor responsible for the specificity of cell damage by the Parkinsonism-causing neurotoxin MPTP. It will be shown that these metals in vitro, and MPTP, each potentiate the autoxidation of dopamine and the production of aminochrome through the generation of superoxide, hydroxyl radicals, hydrogen peroxide and reactive semiquinones. Moreover, the same metals contribute to the oxidation of MPTP itself, further enhancing dopamine autoxidation. PMID- 2985074 TI - Leukotriene C4 formation catalyzed by three distinct forms of human cytosolic glutathione transferase. AB - The ability of three distinct types of human cytosolic glutathione transferase to catalyze the formation of leukotriene C4 from glutathione and leukotriene A4 has been demonstrated. The near-neutral transferase (mu) was the most efficient enzyme with Vmax= 180 nmol X min-1 X mg-1 and Km= 160 microM. The Vmax and Km values for the basic (alpha-epsilon) and the acidic (pi) transferases were 66 and 24 nmol X min-1 X mg-1 and 130 and 190 microM, respectively. The synthetic methyl ester derivative of leukotriene A4 was somewhat more active as a substrate for all the three forms of the enzyme. PMID- 2985075 TI - Activation of NADPH oxidase in human neutrophils. Synergism between fMLP and the neutrophil products PAF and LTB4. AB - The induction of the respiratory burst in human neutrophils by combinations of fMLP and either PAF or LTB4 was studied. Pretreatment with PAF (0.0001 to 10 uM), which by itself did not elicit the burst, greatly enhanced the rate and extent of fMLP-induced superoxide production. A synergism of a different kind was observed with the reversed stimulus sequence: Pretreatment with fMLP made the neutrophils capable to respond to PAF with superoxide production. A moderate enhancement of the fMLP response was also obtained following pretreatment with LTB4. The response of the cells to LTB4, however, was not influenced by fMLP, and no synergism was observed between the two neutrophil products PAF and LTB4. The results of this study demonstrate a marked synergism between fMLP and PAF and suggest that PAF may function as an amplifier of the respiratory burst response of stimulated neutrophils. PMID- 2985076 TI - Presence of a 1,25-dihydroxy-vitamin D3 receptor in chick skeletal muscle myoblasts. AB - The presence of a specific receptor for 1,25-dihydroxy-vitamin D3 was investigated in myoblasts released from chick embryo skeletal muscle by trypsin and collagenase treatment. Density gradient analysis of the cytosol obtained from these muscle cell preparations showed that 1,25-dihydroxy-vitamin D3 binds specifically to a 3.7 S macromolecule. Scatchard analysis yielded an equilibrium dissociation constant of 2.46 x 10(-10) M and a Nmax of 74 fmol/mg of cytosol protein. The data is in agreement with previous evidence which indicates that the action of the vitamin D metabolite on muscle Ca uptake is mediated by de novo protein and RNA synthesis, and supports the concept that muscle is a target organ for 1,25-dihydroxy-vitamin D3. PMID- 2985077 TI - Binding to erythrocyte membrane is the physiological mechanism for activation of Ca2+-dependent neutral proteinase. AB - In the presence of micromolar concentrations of Ca2+ the catalytic 80 kDa subunit of human erythrocyte procalpain binds to the cytosolic surface of the erythrocyte membrane. Binding is rapid, highly specific and is reversed by the removal of Ca2+. In the bound form the 80 kDa catalytic subunit undergoes a rapid conversion to calpain, the active 75 kDa Ca2+-requiring proteinase. The activated proteinase produces extensive degradation of membrane components, particularly of band 4.1 and 2.1 proteins. Binding to membranes may represent an obligatory physiological mechanism for the conversion of procalpain to calpain. PMID- 2985078 TI - Corticotropin-releasing factor: immunohistochemical colocalization with adrenocorticotropin and beta-endorphin, but not with Met-enkephalin, in subpopulations of duodenal perikarya of rat. AB - By the use of two different double-staining techniques (simultaneous staining of adjacent serial sections and the double-staining elution method) it was possible to demonstrate that a corticotropin-releasing factor (CRF) immunofluorescence co existed with an adrenocorticotropin (ACTH) and beta-endorphin (beta-END) immunoreactivity, but not with a Met-enkephalin (Met-ENK) immunostaining, within perikarya subpopulations of both the myenteric and submucousal plexus of the rat duodenum. Not a single Met-ENK-positive neuronal cell body was stained also for CRF, ACTH or beta-END. Even nerve fibres, localized in both the myenteric plexus and closely to submucousal blood vessels (probably arterioles), revealed a CRF immunofluorescence, which is also colocalized with an beta-END staining. These results are quite different to the recent observations in the mammalian hypothalamus, suggesting that some myenteric and submucousal plexus neurons may synthesize CRF as well as beta-END and ACTH, but not Met-ENK. The colocalized peptides might be concomitantly released into the synaptic cleft after terminal stimulation. PMID- 2985079 TI - Human testicular angiotensin-converting enzyme is a mixture of two molecular weight forms. Only one is similar to the seminal plasma enzyme. AB - Two molecular weight (Mr) forms of angiotensin-converting enzyme are present in human testis. Both the high Mr 140,000 form and the low Mr 90,000 form are catalytically similar but immunologically distinct. After isoelectric focusing, the profile of sialylated Mr 140,000 isozymes resembled that of seminal plasma converting enzyme, whereas the nonsialylated Mr 90,000 isozymes were distinct. These data suggest that the Mr 140,000 testicular converting enzyme may be a source of converting enzyme in seminal plasma. PMID- 2985080 TI - 2-((2-Pyridylmethyl)sulfinyl)benzimidazoles: acid sensitive suicide inhibitors of the proton transport system in the parietal cell. AB - 2-((2-Pyridylmethyl)sulfinyl)benzimidazoles, selective inhibitors of the H+/K+ ATPase in the parietal cells of the stomach, have been investigated concerning their chemical behaviour in acidic medium. Protonation of the sulfoxide and subsequent elimination of water forms a sulfenium ion or a chemical equivalent thereof. If no external nucleophiles are present, a rearrangement process takes place. In the presence of mercaptans, the sulfenium ion is trapped giving rise to a variety of products. On the basis of these results, a mechanistic scheme is proposed for the inactivation of the H+/K+-ATPase by these compounds. PMID- 2985081 TI - Effect of cyclic AMP and cyclic GMP on the autophosphorylation of elongation factor 1 from wheat embryos. AB - Extensively purified EF-1H (EF-1 alpha beta beta' gamma) from wheat embryos had a protein kinase activity and phosphorylated EF-1 beta which is one of the two EF Ts-like factors (EF-1 beta and EF-1 beta'). In this reaction ATP and GTP were equally effective as phosphate donors, and threonine residue was phosphorylated. At 10(-7)M, 3', 5' cyclic GMP stimulated both the phosphorylation and phe-tRNA binding reactions, whereas 3', 5' cyclic AMP inhibited both reactions. These findings indicate that phosphorylation of EF-1H may play an important role in the translational control of protein biosynthesis at the elongation step. PMID- 2985082 TI - Why is a specific amino acid sequence of F glycoprotein required for the membrane fusion reaction between envelope of HVJ (Sendai virus) and target cell membranes? AB - Since homology among the fusogenic sequences of the F glycoprotein of paramyxoviruses is high compared with other hydrophobic sequences such as transmembrane sequences and signal peptides, specific binding of this segment to some membrane component was suspected. Thus, temperature-dependent binding of cholesterol to F protein was found. A virus inhibitory peptide (Z-D-Phe-L-Phe Gly) was found to bind to virus particles also temperature dependently. These dependency are very similar to that of the fusion reaction. Binding of cholesterol and the peptide to the fusogenic sequence of F protein was suggested based on construction of molecular models. Importance of these bindings for understanding the fusion mechanism is discussed. PMID- 2985083 TI - Nucleosidediphosphate-kinase induction by human recombinant IL-2 in mouse NK cells. AB - The ability of human recombinant IL-2 to induce NDP-kinase in mouse NK cells has been studied. A significant increase in the amount of NDP-kinase was observed when the cells were exposed to IL-2 (100 units/ml) for 3 h at 37 degrees C. The enzyme inducting ability of human recombinant IL-2 was similar to that of native mouse IL-2 in the cells. The enzymatic characteristics [chemical requirements for the phosphoenzyme formation and molecular size of two distinct subunits (18,000 and 20,000 daltons)] of NDP-kinase from IL-2 treated cells were similar to those of the enzymes from EAT cells. The enzyme's biological role in the initiation of cell proliferation by IL-2 has been discussed. PMID- 2985084 TI - Effects of caerulein-related peptides on cholecystokinin receptor bindings in brain and pancreas. AB - A number of caerulein (CLN)-related peptides were synthesized and compared in terms of their affinities for cholecystokinin (CCK) receptors. We have found that these peptides can be classified into three types according to their relative affinities for the brain and pancreatic receptors. The first group (type A) of peptides includes CLN and analogs retaining the Tyr(SO3H)4 residue and the COOH terminal amide group. Type A peptides were as potent as CLN in inhibiting [125I]BH-CCK-8 binding and showed almost the same affinities for pancreatic and brain receptors. When the Tyr(SO3H)4 residue was either deleted or desulfated (type B), the affinities of the peptides decreased remarkably for the pancreatic receptors but much less for the brain receptors. The type C peptides were deamidated, oxidized, or shortened in the COOH-terminal region and exhibited greatly decreased affinities for both brain and pancreatic receptors but a much greater decrease for the brain receptor. These results indicate that, although the Tyr(SO3H)4 residue and the COOH-terminal structure are both essential for CLN to bind to the CCK receptors, the former is of critical importance for the binding to the pancreas and the latter is rather important for the binding to the brain. PMID- 2985085 TI - Spin-trapping studies on the effects of vitamin E and glutathione on free radical production induced by 3-methylindole. PMID- 2985086 TI - Opiate receptor binding profile in the rabbit cerebellum and brain membranes. AB - The equilibrium dissociation constants (Kd) and maximal binding capacities of tritiated dihydromorphine (DHM), [D-Ala2,D-Leu5]enkephalin (DADLE), ethylketocyclazocine (EKC) and human beta-endorphin (beta-EP) in rabbit cerebellum and brain membranes have been investigated. Binding of tritiated DHM and DADLE was adequately described by a single affinity class of binding sites, while that of EKC required two affinity sites. Binding of tritiated beta-EP was also consistent with a single affinity class of binding sites by Scatchard analysis, but inhibition of the binding with type selective opiate receptor ligands revealed multiple sites. Sequential displacement of a broad spectrum opiate ligand, diprenorphine (DIP), by type selective ligands showed that cerebellum membranes are relatively rich in mu (40%) and deficient in K (12%) binding sites, while brain membranes are relatively rich in K (32%) and deficient in mu (12%) binding sites. beta-EP displaces 88 and 73% of tritiated DIP from cerebellum and brain membranes, respectively, suggesting multiple sites of beta EP binding. PMID- 2985087 TI - Inhibition by free radical scavengers and by cyclooxygenase inhibitors of the effect of acidosis on calcium transport by masseter muscle sarcoplasmic reticulum. AB - In vitro, arachidonic acid depressed calcium transport by sarcoplasmic reticulum (SR) in the homogenate of canine masseter muscle. This effect was inhibited by superoxide dismutase (SOD), a scavenger of the superoxide anion radial ( . O-2), at pH 7.0, and by SOD plus d-mannitol, a scavenger of hydroxyl free radical ( . OH), at pH 5.5. Indomethacin and 2-aminomethyl-4-tert-butyl-6-propionyl phenol (ONO-3144), a compound known to accelerate the conversion of prostaglandin G2 (PGG2) to PGH2 and scavenge free radicals, inhibited the effect of arachidonic acid at both pH 7.0 and pH 5.5. PGG2, but not PGH2, duplicated the effect of arachidonic acid. The effect of PGG2 on SR function was similar to that of exogenous free radicals generated from the xanthine-xanthine oxidase system. Incubation at pH 5.5, in the absence of an exogenous free-radical generating system, depressed SR calcium transport in the homogenate and in isolated SR. This effect in the homogenate was inhibited by indomethacin or by ONO-3144. At 10-min incubation at pH 5.5, SOD partially and temporarily reversed the depressant effect of acidosis. The addition of SOD plus d-mannitol completely reversed the system. d-Mannitol alone was ineffective. Arachidonic acid was able to mimic these effects of acidosis, except that arachidonic acid further depressed isolated SR calcium transport. These results demonstrate that acidosis can depress SR calcium transport in the homogenate of masseter muscle by an oxygen free radical mechanism by the generation of . O-2 and . OH. Our results also demonstrate that significant oxygen radical generation can occur through the cyclooxygenase pathway of arachidonic acid metabolism at an acidotic pH in the cellular environment outside of the SR of the muscle cell, and seems to be responsible for the generation of the . OH derived from . O-2. PMID- 2985088 TI - [Localization of mutation leading to resistance of E. coli RNA polymerase to the antibiotic streptolydigin in the gene rpoB coding for the beta-subunit of the enzyme]. AB - For the first time a mutation of streptolydigin resistance was localized. It was discovered to be a double substitution, namely Gly544----Asp, Phe545----Ser, in the region where most rif-r mutations are located. One may suppose that this region takes part in the formation of both elongation NTP binding site, blocked by streptolydigin, and RNA chain binding and translocation site that is blocked by rifampicin. PMID- 2985089 TI - [Synthesis and biological activity of analogs of a nonapeptide inhibitor of peptidyldipeptidase]. AB - The synthesis of 5 analogues of the effective inhibitor of peptidyl dipeptidase, teprotide, has been carried out. The inhibitory and bradykinin-potentiating activity of these compounds has been assayed. N-Terminal pyroglutamic acid and positive charge of arginine in position 4 were found to be essential for biological activity of the inhibitor. PMID- 2985090 TI - Cartilage nucleoside triphosphate pyrophosphohydrolase. II. Role in extracellular pyrophosphate generation and nucleotide metabolism. AB - Extracellular generation of inorganic pyrophosphate (PPi) in cartilage organ culture is markedly augmented by ATP.ATP, not an ATP metabolite (ADP, AMP, adenosine) is necessary for this augmentation. Excess PPi production is effectively blocked by known inhibitors of nucleoside triphosphate (NTP) pyrophosphohydrolase (EDTA, EGTA, dithiothreitol). Excess 32P-PPi is generated directly from gamma 32P-ATP by cartilage, as substrate and product have similar specific activities. These findings strongly favor ecto-NTP pyrophosphohydrolase as the source of extracellular PPi generation in the presence of NTP. Additionally, active nucleotide and nucleoside catabolism is demonstrated in these cartilage organ cultures. PMID- 2985091 TI - A persistent adenovirus type 1 infection in synovial tissue from an immunodeficient patient with chronic, rheumatoid-like polyarthritis. PMID- 2985092 TI - Angiotensin converting enzyme inhibitors and rheumatoid arthritis. PMID- 2985093 TI - [The pharmacologic effect of flupirtine, a structurally new analgesic]. AB - The analgesic potency of ethyl-N-[2-amino-6-(4-fluorophenylmethylamino)pyridin-3 yl]carb ama te (flupirtine, D 9998) in mice and rats in Haffner's test, electro pain test and Randall-Selitto test (inflammation induced pain) lies between the more potent dextromoramide and methadone and the more weakly active pethidine, dextropropoxyphene, codeine, phenacetin and paracetamol. In comparison to codeine flupirtine is up to 4 times more potent, up to 2 times more active than pethidine and 4 times more potent than dextropropoxyphene in the above-mentioned methods. With one exception of inflammation induced pain, where flupirtine shows an activity of about 1 1/2 times that of phenacetin and paracetamol, both analgesics are about 10 to nearly 30 times less active than flupirtine in other above mentioned tests. In the hot plate test flupirtine is twice as active as codeine and approximately 10 times more active than phenacetin and paracetamol. The weakest analgesic activity of flupirtine is seen in acetic acid test where it is about half as active as codein and approximately as active as dextropropoxyphene. Nevertheless, flupirtine is up to 10 times more potent than phenacetin and paracetamol. The acetic acid test is claimed to be non-specific according to our own experience and to other authors. Flupirtine is enterally absorbed at a higher degree than the other tested centrally acting analgesics. In regard to the results of various analgesic investigations in mice and rats flupirtine can be classified as a medium to strong acting analgesic. The duration of action of flupirtine is comparable to that of codeine. Experiments with flupirtine suggest that there are some convincing criteria for a pronounced central acting component of its analgesic activity. These criteria are the strong efficacy in the hot plate and Haffner's test, in which only centrally acting analgesics show distinct effects, and the finding that flupirtine increases the pain threshold for vocalisation in rats and mice excluding a pure reflex of the spinal cord. In current experiments concerning the mode of action flupirtine exhibits a distinct central analgesic component of action. In spite of its relatively high analgesic potency which corresponds to that of opiates flupirtine does not show any other signs of opiate properties and other potent analgesics. Thus, flupirtine does not develop tolerance in mice and rats after 19 or 17 days of daily administration.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985094 TI - Correlation between octanol solvent distribution ratio and liquid chromatographic retention for some 7- and 9-substituted 1-methyl-isoguanines. AB - Octanol/aqueous buffer solvent distribution ratio and liquid chromatographic retention in a primarily aqueous phase were determined and correlated to estimated substituent Hansch pi values for a series of 1-methyl-isoguanines. Substituent connectivity values (simple, valence, and bond length) were also computed and factored into the correlation equation between known in vitro activity data and these lipophilic parameters. The liquid chromatographic retention data were best correlated with measured activity accounting for 61% of the variation while distribution ratio and Hansch pi accounted for 56% or 52%, respectively, when each was considered alone. Valence connectivity accounted for 30% of the variation alone but did not add significantly to the equations when factored with the other parameters. PMID- 2985095 TI - Inhibition of [3H]clonidine binding to rat brain alpha-adrenoceptors by imidazolidines. Correlation of binding potency and hypotensive activity. AB - The inhibition of specifically bound [3H]clonidine (spec. activity 26.7 Ci/mmol; final concentration 0.4 nmol/l) to rat brain membranes was determined for clonidine (2-[2,6-dichlorophenylimino]-2-imidazolidine hydrochloride, Catapresan) and a series of 28 congeneric imidazolidines with different substituents attached to the phenyl moiety and to the bridge nitrogen. The affinity of the compounds for the binding sites labelled by [3H]clonidine greatly depended on these substituents. The potencies of the substances in vitro to inhibit [3H]clonidine binding were correlated with reported hypotensive activities in vivo obtained in anaesthetized rabbits and normotensive rats. Correlation coefficients of r = 0.92 and of r = 0.83, resp., resulted which could be improved upon inclusion of lipophilicity omitting substances with logarithms of octanol/buffer partition coefficient less than -0.5. This finding was explained by the difference in accessibility to the central alpha-adrenoceptor binding sites in vitro and in vivo. Whereas no penetration barrier of importance will affect the process of in vitro binding, the blood/brain barrier has to be crossed before a hypotensive effect can be initiated. The binding affinities of the imidazolidines for central sites identified by [3H]clonidine may provide an indication for potential blood pressure lowering efficacy. PMID- 2985096 TI - Differentiating aspects of opioid receptor binding by [3H](-) (1R,5R,9R,2''S)-5,9 dimethyl-2-tetrahydrofurfuryl-2'-hydroxy-6,7- benzomorphan hydrochloride ([3H]Mr 2034), a drug preferentially acting on kappa-receptors. AB - Receptor binding experiments with masking of mu- and delta-receptors in the presence of an excess of unlabelled dihydromorphine and [D-Ala2, D Leu5]enkephalin have been carried out. They indicate that 12 to 17% of original high affinity binding of [3H]Mr 2034 [-)(1R,5R,9R,2"S)-5,9-dimethyl-2 tetrahydrofurfuryl-2'-hydroxy-6, 7-benzomorphan hydrochloride), an opioid kappa agonist, could then be detected as kappa-receptor sites in brain membranes both from untreated rats and from rats pretreated with naloxazone, too. Because of the irreversible blockade of the high affinity binding sites by naloxazone (naloxone hydrazone) treatment, the masking effects of mu- and delta-selective ligands seem to be mediated by the low affinity binding sites of these opioid receptor types. As could be shown before with [3H]Mr 2034, another kappa-agonist, [3H]bremazocine does not seem to be affected in its binding properties by naloxazone treatment of the rat in vivo. Displacement studies with several opioid agonists and antagonists, [3H]Mr 2034 and [3H]ethylketocyclazocine as radioligands in brain membranes from naloxazone treated rats and untreated controls provided further support for the evidence of two different kappa-receptors. PMID- 2985097 TI - Acquired immuno deficiency syndrome (AIDS): experience in the Puerto Rico Medical Center. PMID- 2985098 TI - Synthesis and processing of human serum apolipoprotein AII in vitro and in Hep G2 cells. AB - The synthesis and structure of the primary translation product of apo AII in a human liver poly(A+) mRNA primed cell-free system and its cotranslational modification was studied parallel to studies in vivo with Hep G2 cells, a human hepatoma cell line. The primary translation product is a preproprotein containing 100 amino acid residues, which is cleaved by the signal peptidase of endoplasmic reticulum to pro-apo AII with the loss of the N-terminal pre-sequence consisting of 18 amino acid residues. Hep G2 cells contain about equal amounts of the proform of apolipoprotein AII and of mature apo AII. Approximately in the same ratio pro- and mature apo AII are secreted into the medium. Determination of the partial amino-acid sequence by automated Edman degradation of the labelled prepro and proforms of apo AII led to the segmentation of the N-terminus of the primary translation product, consisting of 23 amino acid residues, into the pre-sequence (18 residues) and the pro-sequence (5 residues) with terminal Arg-Arg-residues at the cleavage site to apo AII. We must therefore correct our previously postulated 17 and 6 residues containing segmentation. So far no information has been obtained in which compartment and at what stage of posttranslational events the dimerization occurs by formation of the single disulfide bond at position Cys6 in the mature apo AII structure, leading to the symmetrical molecule. PMID- 2985099 TI - Amphiphilic cationic drugs and phospholipids influence the activities of beta galactosidase and beta-glucosidase from liver lysosomal fraction of untreated rats. AB - The influence of amphiphilic drugs and phospholipids on the activities of beta galactosidase and beta-glucosidase from liver lysosomal fractions of untreated rats, isolated by affinity chromatography using castor bean lectins, was studied in vitro. Chloroquine (93 microM) inhibited beta-galactosidase activity by about 30%, while O,O'-bis(diethylaminoethyl)hexestrol showed no inhibitory effect. Neutral phospholipids (phosphatidylcholine, phosphatidylethanolamine, sphingomyelin) inhibited the enzyme slightly, while the enzyme activity was drastically reduced in the presence of acidic phospholipids (phosphatidylinositol, phosphatidylserine, bis-(monoacylglycero)phosphate). Lysosomal beta-glucosidase was strongly inhibited by chloroquine and O,O' bis(diethylaminoethyl)-hexestrol. The neutral phospholipids showed only a moderate inhibitory effect, whereas the acidic phospholipids were stimulators. Bis(monoacylglycero)phosphate was by far the best stimulating compound. PMID- 2985100 TI - The effect of unprocessed wheat bran on blood glucose and plasma immunoreactive insulin levels during oral glucose tolerance test in obese children. AB - Blood glucose and plasma immunoreactive insulin concentrations were measured during oral glucose tolerance test in 10 obese children. Oral glucose was given by itself or combined with 15 g unprocessed wheat bran. Bran significantly reduced the blood glucose and plasma immunoreactive insulin concentrations at 30 min of the tolerance test. It is concluded that supplementation of obese children's diet with unprocessed bran is advantageous. PMID- 2985101 TI - Ethnic differences in human lymphocytic cyclic AMP production after isoprenaline stimulation and propranolol blockade. AB - The stimulatory effects of isoprenaline on human lymphocytic cyclic AMP (cAMP) and blockade by propranolol were studied in vitro in healthy Black and White volunteers. Basal levels of lymphocytic cAMP were significantly higher in Blacks than in Whites. Stimulation with isoprenaline caused a dose-related increase in cAMP, which was in concentrations of 10(-9) to 10(-5)M significantly greater in Blacks than in Whites. Blockade by 10(-4)M propranolol did not affect basal cAMP levels significantly, but increases in cAMP levels were significantly smaller in both groups after 10(-9) to 10(-2)M isoprenaline, while differences between cAMP levels in Blacks and Whites were still significant at concentrations 10(-9) to 10(-3)M. The increased cAMP concentration in lymphocytes of Blacks probably reflects a higher degree of beta 2-adrenoceptor activity which could be due to either a greater number and/or greater sensitivity of lymphocytic beta 2 adrenoceptors in Blacks than in Whites. PMID- 2985102 TI - The effect of dietary supplementation with linoleic and gammalinolenic acids on the pressor response to angiotensin II--a possible role in pregnancy-induced hypertension? AB - Dietary supplementation with 3 g/day linoleic acid, 32 mg/day gammalinolenic acid and co-factors for prostaglandin synthesis was given to 10 pregnant and 10 non pregnant subjects for a week. Their pressor response to the infusion of three doses of angiotensin II (AII) (pregnant: 4, 8, 16 ng kg-1 min-1: non-pregnant: 1, 2, 4 ng kg-1 min-1) was then compared with that of 40 pregnant and 24 non pregnant controls who had not been given such supplementation. Dietary supplementation was not associated with changes in basal systolic or diastolic blood pressure or heart rate during the week of treatment in pregnant or non pregnant subjects. Basal systolic blood pressure, diastolic blood pressure and heart rate values did not differ between the treated and untreated subjects in each group. The diastolic pressor response to AII was significantly less after treatment at all doses; the effect was more marked in the pregnant subjects. The systolic response to AII, normally less than the diastolic, was somewhat blunted in the treated pregnant patients at the two higher infusion doses. No significant effect was found in the non-pregnant group. Evidence from other studies suggests that increasing plasma linoleic acid concentrations leads to increased plasma concentrations of epoprostenol (prostacyclin, PGI2) while increased availability of gammalinolenic acid is associated with a rise in prostaglandin E1 and E2 production. Pregnancy-induced hypertension is associated with a diminished tissue production of both epoprostenol and E series prostaglandins, and with an enhanced pressor response to AII.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985104 TI - Bronchioloalveolar cell carcinoma: a 21-year retrospective study of cases at the Western Infirmary, Glasgow. AB - A 21-year retrospective autopsy study of bronchioloalveolar cell carcinoma in the Western District of Glasgow is presented which includes a detailed review of the case notes and post-mortem pathology of 18 patients. Aspects of the histogenesis of the tumour and its prognosis are included in the discussion. It is concluded that bronchioloalveolar cell carcinoma is a valid term which describes a disease entity that is amenable to successful surgical resection when at a limited stage but that in pathological terms it represents a heterogeneous population of tumours. PMID- 2985103 TI - Effects of repeated doses of enalapril on renal function in man. AB - Ten healthy subjects received 10 mg oral enalapril (MK 421) daily for a period of 8 days. Renal clearances of electrolytes, urate and phosphate were monitored and factored for glomerular filtration rate, as measured by creatinine clearance, with particular emphasis on the first and eighth day of treatment. Apart from a fall of around 10% in creatinine clearance between 1-2 h on both days 1 and 8, GFR remained unchanged throughout the study. Fractional sodium excretion increased in a biphasic manner by approximately 50% over control between 1-2 h and 4-8 h on day 1. Significant chloruresis (+39.0 +/- 12.9%) and kaluresis (+26.5 +/- 10.3%) occurred between 4-8 h. Urinary pH increased between 0-1 h (+0.29 +/- 0.12; P less than 0.05), and between 4-8 h (+0.50 +/- 0.08; P less than 0.01). The biphasic saluretic effect was also seen between 1-2 h and 4-8 h on day 8. Enalapril caused significant increases in urate and phosphate excretion on day 8 of therapy. There was a biphasic increase in fractional urate excretion at 1-2 h (+28.1 +/- 6.9%; P less than 0.05) and at 4-8 h (+21.0 +/- 6.0% P less than 0.01). Significant phosphaturia (+36.8 +/- 5.2%; P less than 0.05) was also observed at 4-8 h on day 8. Urinary drug excretion was also biphasic; over the first 2 h the predominant drug form was unchanged enalapril, whilst the peak excretion of the diacid metabolite, enalaprilat, occurred at 4-8 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985105 TI - Upper abdominal lymphadenopathy as first presentation of relapse, identified by ultrasonography, in patients treated for small cell (oat cell) bronchogenic carcinoma. AB - Ten patients treated for 'limited' stage small cell lung cancer presented during follow-up with epigastric or back pain together with nausea and/or vomiting. Upper abdominal lymphadenopathy was shown to be the first symptomatic site of relapse. A single fraction of radiotherapy provided a convenient, effective form of palliation. Ultrasound scanning proved an accurate, readily available technique for the diagnosis of this form of intra-abdominal disease. PMID- 2985106 TI - Nitric acid-induced injury in the hamster lung. AB - Severe airway lesions can result following exposure to various pathogens or toxic gases and can show a variety of pathologic lesions including necrotizing bronchitis, bronchiolitis and bronchiectasis. The purpose of this study was to develop a chronic airway lesion in the hamster, a species recognized for its lack of endogenous pulmonary disease. We have successfully adapted a technique of inducing rabbit airway lesions with nitric acid to the hamster lung and have characterized the morphologic, morphometric and biochemical features of the model. Following a transorotracheal instillation of 0.5% HNO3 in saline, Syrian golden hamsters showed during a 60-day study period a spectrum of airway changes including acute bronchitis, acute bronchiolitis, obliterative bronchiolitis, bronchiolectasia and bronchiectasis. Morphometric changes in the HNO3-treated hamsters included decreased lung volumes and decreased internal surface areas. Biochemical changes showed increases in lung weight and in total collagen and elastin. The model is useful because a prolonged insult to the airways develops rapidly and persists over a long period of time, important features for investigations designed to study the effects of superimposed insults upon pre existent airway lesions. PMID- 2985107 TI - Lactoferrin in the myeloproliferative disorders: a search for granulopoietic regulator defects. AB - Evidence of a defect in the negative feedback control of granulopoiesis in the myeloproliferative disorders is presented. Neutrophil release of lactoferrin, plasma lactoferrin concentration, the number of lactoferrin receptor sites on mononuclear cells and granulocyte-monocyte colony stimulating factor (GM-CSF) release from peripheral blood cells were determined in patients with myeloproliferative disorders and compared with normal individuals. There was a significantly (P less than 0.001) decreased release of lactoferrin from the neutrophils of patients with myeloproliferative disorders which could be responsible for a reduced suppression of GM-CSF release from mononuclear cells which in turn may be responsible for the increased myeloid proliferative activity in patients with myeloproliferative disorders. PMID- 2985108 TI - CNS relapse in children with acute lymphoblastic leukaemia. PMID- 2985109 TI - Paget's disease of the vulva complicated by bladder carcinoma. Case report. PMID- 2985110 TI - Flavocytochromes c: transient kinetics of photoreduction by flavin analogues. AB - Kinetics of reduction of phototrophic bacterial flavocytochromes c by exogenous flavin semiquinones and fully reduced flavins generated by laser flash photolysis have been studied. The mechanisms of reduction of Chromatium and Chlorobium flavocytochromes c are more similar to one another than previously thought. Neither protein is very reactive with neutral flavin semiquinones (k less than 10(7) M-1 s-1), and the reactions with fully reduced flavins are slower than expected on the basis of comparison with other electron-transfer proteins of similar redox potentials. Deazaflavin radical is reactive with the flavocytochromes c by virtue of its low redox potential, but this reaction is also slower than expected on the basis of comparison with other electron-transfer proteins. These experiments indicate that the active site for reduction of flavocytochrome c is relatively buried and probably inaccessible to solvent. Fully reduced FMN does not show an ionic strength effect in its reaction with flavocytochrome c, which demonstrates that the active site is uncharged. Sulfite, which forms an adduct with protein-bound FAD, partially blocks heme reduction. This shows that heme is reduced via the FAD. The rate constant for intramolecular electron transfer between FAD and heme must be on the order of 10(4) s-1 or larger. PMID- 2985111 TI - Extracellular cGMP phosphodiesterase related to the rod outer segment phosphodiesterase isolated from bovine and monkey retinas. AB - A phosphodiesterase (PDE) has been characterized in the interphotoreceptor matrix (IPM) of light-adapted fresh bovine retinas. It is obtained through a gentle rinsing of the retinal surface under conditions where the light-activated rod outer segment (ROS) enzyme remains attached. The enzyme has an apparent native molecular weight of 350 000 by gel filtration and appears as a doublet at Mr 47 000 and 45 000 on sodium dodecyl sulfate-polyacrylamide gels. It has an apparent Km value for cGMP of 33 microM and an apparent Km value for cAMP of 2200 microM. It is activated 3-6-fold by protamine and over 40-fold by trypsin. Protamine has no effect on the Km for cGMP while trypsin decreases the Km for cGMP by a factor of 2. The enzyme occurs in at least two forms as evidenced by two distinct peaks of activity after gel electrophoresis under nondenaturing conditions. A heat stable inhibitor is tightly bound to the enzyme. The inhibitor obtained from the IPM PDE inhibits 98% of the activity of the trypsin-activated ROS PDE: conversely, the inhibitor obtained by boiling the ROS PDE completely inhibits the trypsin-activated IPM enzyme. A high-affinity monoclonal antibody to the active site of the ROS PDE, ROS 1 [Hurwitz, R., Bunt-Milan, A.H., & Beavo, J. (1984) J. Biol. Chem. 259, 8612-8618], quantitatively absorbs the IPM PDE. These observations indicate a clear relationship between these two PDEs even though their location, sizes, and specific functions in the retina appear to be distinct. PMID- 2985112 TI - Spin-label studies of lipid-protein interactions in (Na+,K+)-ATPase membranes from rectal glands of Squalus acanthias. AB - Lipid-protein interactions in (Na+,K+)-ATPase-rich membranes from the rectal gland of Squalus acanthias have been studied by using spin-labeled lipids in conjunction with electron spin resonance (ESR) spectroscopy. Lipid-protein associations are revealed by the presence of a second component in the ESR spectra of the membranes in addition to a component which corresponds very closely to the ESR spectra obtained from dispersions of the extracted membrane lipids. This second component corresponds to spin-labeled lipids whose motion is very significantly restricted relative to that of the fluid lipids in the membrane or the lipid extract. A stoichiometry of approximately 66 lipids per 265 000-dalton protein is found for the motionally restricted component of those spin labeled lipids (e.g., phosphatidylcholine) which show least specificity for the protein. This corresponds approximately to the number of lipids which may be accommodated within the first shell around the alpha 2 beta 2 protein dimer. A selectivity of the various spin-labeled lipids for the motionally restricted component associated with the protein is found in the following order: cardiolipin greater than phosphatidylserine approximately stearic acid greater than or equal to phosphatidic acid greater than phosphatidylglycerol approximately phosphatidylcholine approximately phosphatidylethanolamine approximately androstanol. PMID- 2985113 TI - Hierarchy of binding sites for chromosomal proteins HMG 1 and 2 in supercoiled deoxyribonucleic acid. AB - The interaction of chromosomal proteins HMG 1 and 2 with various DNA structures has been examined with plasmid pPst-0.9, which contains DNA sequences that can form the Z-DNA conformation and palindromic sequences that can form cruciform structures. Direct binding and competition experiments with 32P-labeled plasmid indicated that proteins HMG 1 and 2 preferentially bind to supercoiled form I DNA as compared to double-stranded linear DNA. The preferential binding to form I is due to the presence of single-stranded regions in this DNA. The binding of HMG 1 and 2 to the form I plasmid results in inhibition of S1 nuclease digestion in a selective manner. The B-Z junction is preferentially protected as compared to the cruciform, which in turn is more protected than other minor S1-sensitive structures present in pPst-0.9. Our results indicate that the binding of HMG 1 and 2 proteins to DNA is not random in that HMG 1 and 2 can distinguish between various S1 nuclease sensitive sites in the plasmid. The existence of a hierarchy of DNA binding sites for these proteins suggests that they can selectively affect the structure of distinct regions in the genome. PMID- 2985114 TI - Direct photoaffinity labeling of tubulin with guanosine 5'-triphosphate. AB - Irradiation of tubulin in the presence of [3H]GTP or [3H]GDP at 254 nm led to the covalent incorporation of nucleotide into the protein. The specific nature of the labeling was shown in the following manner: with tubulin depleted of exchangeable nucleotide, the amount of labeling increased to a plateau value as the [3H]GTP concentration was increased, with saturation being reached at a ratio of approximately 1.5; the same amount of labeling was obtained with GTP/tubulin ratios of 1 and 100; [3H]GMP was not incorporated into the dimer, nor did GMP inhibit the incorporation of [3H]GTP; [3H]ATP was not incorporated; [3H]GTP incorporation did not occur into denatured tubulin or into serum albumin. When [alpha-32P]GTP was used in the irradiation experiments, sodium dodecyl sulfate polyacrylamide gel electrophoresis of the carboxymethylated protein demonstrated that the incorporated label was associated with the beta subunit. The radiation treatment did cause changes in the tubulin molecule resulting in a decrease in assembly competence and in sulfhydryl groups, but these effects were minimized when a large excess of GTP was present during irradiation. Labeling of tubulin in the assembled state was much less than that observed in the free state. PMID- 2985115 TI - The involvement of cytoskeleton in the regulation of transbilayer movement of phospholipids in human blood platelets. AB - Activation of human platelets by different activators resulted in a different extent of degradation of the cytoskeletal proteins actin-binding protein and myosin, as well as of the non-cytoskeletal protein P235. The highest extent of proteolysis was observed with Ca-ionophore A23187 and decreased on going from A23187 greater than collagen plus thrombin greater than collagen greater than thrombin = ADP. The same order of potency has been found previously ((1983) Biochim. Biophys. Acta 736, 57-66) for the ability of platelet activators to induce exposure of aminophospholipids in the outer leaflet of the platelet plasma membrane, and to stimulate platelets to become procoagulant. Degradation of cytoskeletal proteins as a result of platelet stimulation by collagen plus thrombin was prevented in the presence of dibutyryl cAMP or EDTA but not in the presence of aspirin. This also runs in parallel with platelet procoagulant activity. Moreover, platelets from a patient with a partial deficiency in platelet procoagulant activity revealed a diminished extent of degradation of cytoskeletal proteins upon platelet stimulation with collagen plus thrombin. It is concluded that alterations in cytoskeletal organization upon platelet stimulation may lead to alterations in the orientation of (amino)phospholipids in the plasma membrane, and may therefore play a regulatory role in the expression of platelet procoagulant activity. PMID- 2985116 TI - Na+-like effect of imidazole on the phosphorylation of (Na+ + K+)-ATPase. AB - A high basal level of phosphorylation (approx. 70% of the optimal Na+-dependent phosphorylation level) is observed in 50 mM imidazole-HCl (pH 7.0), in the absence of added Na+ and K+ and the presence of 10-100 microM Mg2+. In 50 mM Tris HCl (pH 7.0) the basal level is only 5%, irrespective of the Mg2+ concentration. Nevertheless, imidazole is a less effective activator of phosphorylation than Na+ (Km imidazole-H+ 5.9 mM, Km Na+ 2 mM under comparable conditions). Imidazole activated phosphorylation is strongly pH dependent, being optimal at pH less than or equal to 7 and minimal at pH greater than or equal to 8, while Na+-activated phosphorylation is optimal at pH 7.4. This suggests that imidazole-H+ is the activating species. Imidazole facilitates Na+-stimulated phosphorylation. The Km for Na+ decreases from 0.63 mM at 5 mM imidazole-HCl to 0.21 mM at 50 mM imidazole-HCl (pH 7; 0.1 mM Mg2+ in all cases). Imidazole-activated phosphorylation is more sensitive to inhibition by K+ (I50 = 12.5 microM) than Na+-activated phosphorylation (I50 = 180 microM). Mg2+ antagonizes activation by imidazole-H+ and also inhibition by K+. The Ki value for Mg2+ (approx. 0.3 mM) is the same for the two antagonistic effects. Tris buffer (pH 7.0) inhibits imidazole-activated phosphorylation with an I50 value of 30 mM in 50 mM imidazole HCl (pH 7.0) plus 0.1 mM Mg2+. We conclude that imidazole-H+, but not Tris-H+, can replace Na+ as an activator of ATP-dependent phosphorylation, primarily by shifting the E2----E1 transition to the right, leading to a phosphorylating E1 conformation which is different from that in Tris buffer. PMID- 2985117 TI - Loss of latent activity of liver microsomal membrane enzymes evoked by lipid peroxidation. Studies of nucleoside diphosphatase, glucose-6-phosphatase, and UDP glucuronyltransferase. AB - The effects of lipid peroxidation on latent microsomal enzyme activities were examined in NADPH-reduced microsomes from phenobarbital-pretreated male rats. Lipid peroxidation, stimulated by iron or carbon tetrachloride, was assayed as malondialdehyde formation. Independent of the stimulating agent of lipid peroxidation, latency of microsomal nucleoside diphosphatase activity remained unaffected up to microsomal peroxidation equivalent to the formation of about 12 nmol malondialdehyde/mg microsomal protein. However, above this threshold a close correlation was found between lipid peroxidation and loss of latent enzyme activity. The loss of latency evoked by lipid peroxidation was comparable to the loss of latency attainable by disrupting the microsomal membrane by detergent. Loss of latent enzyme activity produced by lipid peroxidation was also observed for microsomal glucose-6-phosphatase and UDPglucuronyltransferase. In contrast to nucleoside diphosphatase, however, both enzymes were inactivated by lipid peroxidation, as indicated by pronounced decreases of their activities in detergent-treated microsomes. According to the respective optimal oxygen partial pressure (po2) for lipid peroxidation, the iron-mediated effects on enzyme activities were maximal at a po2 of 80 mmHg and the one mediated by carbon tetrachloride at a po2 of 5 mmHg. Under anaerobic conditions no alterations of enzyme activities were detected. These results demonstrate that loss of microsomal latency only occurs when peroxidation of the microsomal membrane has reached a certain extent, and that beyond this threshold lipid peroxidation leads to severe disintegration of the microsomal membrane resulting in a loss of its selective permeability, a damage which should be of pathological consequences for the liver cell. Because of its resistance against lipid peroxidation nucleoside diphosphatase is a well-suited intrinsic microsomal parameter to estimate this effect of lipid peroxidation on the microsomal membrane. PMID- 2985118 TI - Electron spin resonance studies of splenic ferritin and haemosiderin. AB - Preparations of haemosiderin and ferritin isolated from iron-loaded human spleens were studied by electron spin resonance (ESR) spectroscopy at X-band (approx. 9.2 GHz). The spectra were mainly composed of two overlapping, broad features, one extremely anisotropic with its major component occurring at 0.1-0.2 T (feature A), the other nearly isotropic and occurring at around g = 2 (feature B). There is relatively more feature A and less feature B in ferritin than in haemosiderin. Both features originate from the iron oxyhydroxide crystallites of these iron proteins which, due to their small size, are superparamagnetic. Feature B is maximal in small cores or at high temperatures, where superparamagnetic fluctuations average out anisotropic magnetic interactions; feature A is greatest at low temperatures or in large cores, for which such fluctuations are blocked and an ESR spectrum characteristic of a magnetically ordered system is observed. It is concluded that there is no evidence in the ESR spectra for 'loose' protein bound Fe3+ in ferritin or haemosiderin, and that haemosiderin cores are on average smaller than those of ferritin. The relationship of the ESR spectra between these two proteins supports the view that haemosiderin is derived from ferritin. PMID- 2985119 TI - Ionization of tyrosine residues in horse-heart ferricytochrome c and its guanidinated and acetylated-guanidinated derivatives. AB - Spectrophotometric titration curves were obtained at 242 nm for native and fully guanidinated horse-heart ferricytochrome c. The cytochrome c data were fit over the pH range 9-12 (I = 0.35) by a theoretical curve with pK' values of 10.35 and 11.70. The slope of the experimental data increases sharply above pH 12.5 suggesting that two tyrosine residues with pK' values greater than 12.5 are exposed by conformation change. The guanidinated cytochrome c data after correction for the alkaline spin-state transition were fit over the entire pH range 9-13.6 (I = 0.35) by a theoretical curve with pK' values 10.37, 10.78, 11.50, and 13.60. These results along with viscosity measurements indicate that the unfolding transition occurs at higher pH in the guanidinated derivative. N Acetylimidazole was used to acetylate specific tyrosyl groups of guanidinated cytochrome c. Assignments of acetylated tyrosine residues were confirmed by peptide mapping of 14C-labelled derivatives. Spectrophotometric titrations with rapid data acquisition of two monoacetylated derivatives allowed assignments of pK'1 (10.37) to Tyr-67 and pK'4 (13.60) to Tyr-97. The basis for the large differences in acidity and chemical reactivity of these two residues is not obvious from the crystallographic structure and may arise from differences in solvent access due to motions of the polypeptide chain. PMID- 2985120 TI - Quarter field resonance and integer-spin/half-spin interaction in the EPR of Thermus thermophilus ferredoxin. Possible new fingerprints for three iron clusters. AB - We describe two new characteristics of the EPR of the seven-iron containing ferredoxin from Thermus thermophilus. First, the reduced state of the 3Fe center, which has traditionally been considered to be EPR-silent, has been found to exhibit a delta m = 4 transition, which is unique for Fe-S centers. This signal is similar to that of high-spin Fe2+-EDTA and supports the suggestion that the ground electronic state of the 3Fe cluster is S = 2. Second, we have recorded the EPR spectrum of the fully reduced protein at 9 and 15 GHz and found that changes occur in the signal which are consistent with a weak electronic spin-spin interaction between the [4Fe-4S]+ (S = 1/2) and the reduced 3Fe center. A theoretical explanation is given for the observation of interaction signals with constant effective g values. PMID- 2985121 TI - Iron-mediated formation of an oxidized adriamycin free radical. AB - Electron paramagnetic resonance studies are reported which demonstrate that the reduction of Fe3+ to Fe2+ by adriamycin results in the formation of an oxidized adriamycin free radical with an EPR signal at g = 2.004. A transient iron adriamycin free radical complex is also observed at g = 2.34. The free radical is quantitated and its aerobic stability is determined. Observation of the oxidized adriamycin free radical signal confirms that adriamycin donates an electron to the bound Fe3+. In the presence of glutathione the drug-mediated reduction of Fe3+ to Fe2+ is bypassed, and the oxidized adriamycin radical signal is not observed. The oxidized adriamycin radicals and reduced oxygen radicals which are formed are two different mediators, whose relative concentrations could modulate the therapeutic and toxic effects of adriamycin. PMID- 2985122 TI - Potentiometric measurements of stoichiometric and apparent affinity constants of EGTA for protons and divalent ions including calcium. AB - The stoichiometric affinity constants of H+, Ca2+, Mg2+ and Sr+ for the ligand EGTA were determined using a modified version of the pH metric method developed by Moisescu and Pusch (Moisescu, D.G. and Pusch, H. (1975) Pfluegers Arch. 355, 243). The values obtained were slightly higher than those previously published. In addition, the shift in the H+ and Ca2+ stoichiometric constants with ionic strength was found to fit an empirical relationship if the total ionic content of the titration solutions was measured in terms of ionic equivalents, Ie (Johansson, L. (1975) Acta Chem. Scand. A29, 365-373), rather than the formal ionic strength, If. Finally, the apparent affinity of EGTA for Ca2+ ions was measured using an abbreviated form of the titration technique. The measured apparent affinity constant agreed with published results only if calculated with respect to pH (measured) of 7.0, rather than pH (concentration). PMID- 2985123 TI - [Study of the effect of the microenvironment on magnetic resonance parameters of spin-labeled human serum albumin in a 2-mm ESR range]. AB - Basic values of g-tensor and Azz component of HF tensor of two spin labels and spin probe on HSA and nitroxyl radicals HO-15, HO-34 in the solvents of different polarity were measured by 2 mm band ESR of 2 mm range. Magnetic-resonance parameters of liophylized and water-solved spin-labeled HSA were shown to correspond to the parameters of the solvents of the label HO-15 and HO-34 in ethyl alcohol and water. A conclusion was drawn concerning the identity of microenvironment of the nitroxyl fragment of liophylized HSA and frozen solution of the label HO-15 and HO-34 in ethyl alcohol and solvatation of the nitroxyl fragment of spin-labeled HSA and label HO-15 (HO-34) by water molecules. PMID- 2985124 TI - [Psoralen-photosensitized formation of free lipid radicals at 77 degrees K]. AB - It is shown by ESR method that 8-methoxypsoralen (8-MOP) under low temperature UV irradiation has no influence on free radical formation of saturated fatty acids (FA). Under the same conditions the quantum yield of free radical photogeneration for unsaturated FA (oleate and limolenate) after addition of 8-MOP increased 7.5 15 times. The ESR spectrum of free radicals generated after interaction of 8-MOP with polyunsaturated FA, may be presented as superposition of signals-CH-(CH = CH)2-, -CH2CHCH2- and -CH2CH2. The molecular mechanism of photosensitized reactions is discussed. PMID- 2985125 TI - [The sodium cycle--a new type of bacterial energetics]. AB - The literature data and experimental results of the author's laboratory on the role of Na+ in bacterial energetics are reviewed. It was shown that certain bacterial species utilize the transmembrane difference of Na+ electrochemical potentials (delta mu Na+) as a convertible membrane-linked form of energy. The membranes of such bacteria were found to contain delta mu Na+ generators (e. g., decarboxylases of some carboxylic acids of NADH-menaquinone reductase). It was shown that delta mu Na+ formed by these generators may support all the three main types of work of the bacterial cell, i. e., chemical (ATP synthesis), osmotic (substrate accumulation) and mechanical (motility). PMID- 2985126 TI - [Participation of the adenyl cyclase system in estradiol-activated growth of tumors in rat mammary glands]. AB - Early effects of estradiol on the adenylate cyclase system in target tissues were investigated. The proliferative processes in the estradiol-dependent mammary tumours and uterus of the rat were arrested by ovariectomy. Subsequent hormonal treatment resulted in adenylate cyclase activation and the increase of the cAMP level in these tissues, probably to be connected with the initiation of the cell growth. These events were followed by a decrease of the cAMP content necessary for active proliferation of tumour cells. cAMP phosphodiesterase is not responsible for the increase in the cyclic nucleotide content, but provides for its further hydrolysis. The stimulation of the adenylate cyclase system at early steps of estradiol action is typical both for normal and transformed estradiol dependent tissues. PMID- 2985127 TI - [Effect of the redox state of glutathione on acetate kinase activity in E. coli]. AB - Oxidized glutathione inhibits acetate kinase (EC 2.7.2.1) of E. coli. The rate of inactivation depends on ATP concentration. The rate constant for the glutathione induced inhibition is 0.17 min-1, Ki is 4.2 mM (pH 7.2, 25 degrees C). The inhibition of acetate kinase by glutathione is reversible, the equilibrium constant being equal to 4.4 or 0.09 at saturating concentrations of ATP (pH 8.0, 25 degrees C). The physiological level of reduced and oxidized glutathione can modulate the acetate kinase activity in vivo. PMID- 2985128 TI - [Cyclic nucleotide-dependent endogenous phosphorylation of rabbit myometrium membranes]. AB - The membrane-bound protein kinase activity in plasma membranes (PM) and sarcoplasmic reticulum (SR) of rabbit myometrium was revealed, which catalyzes the synthesis of protein phosphoester products. cAMP had no effect on the phosphorylation of membrane substrates by soluble protein kinases I and II as well as by the membrane-bound enzyme of SR. At the same time, cAMP (10(-8) stimulated by 200% the phosphorylation of sarcolemmal components at functional rest (FR). In preparations obtained from pregnant animals, cAMP (10(-8) and 10( 5) M) stimulated the phosphorylation of PM 7- and 3-fold, respectively. cGMP had no effect on the phosphorylation of PM and SR proteins at FR. At 10(-5) and 10( 8) M, cGMP stimulated endogenous phosphorylation of PM and SR 7- and 4-fold, respectively. In pregnancy, the degree of endogenous phosphorylation of PM and SR increased by 70% and 260% as compared to that at FR; the activity of soluble protein kinases decreased two times under these conditions. At FR, the sarcolemmal proteins with Mr 35 000, 57 000, 89 000 and 174 000 underwent phosphorylation. The phosphorylation of the proteins with Mr 35 000 and 57 000 was cAMP-dependent. In pregnant animals sarcolemma, the phosphorylation affected the proteins with Mr 47 000, 57 000 and 174 000 and was cAMP-dependent for the former two proteins and cGMP-dependent for the latter protein. At FR, two SR proteins with Mr 47 000 and 168 000, while in pregnant animals the proteins with Mr 47 000, 132 000 and 168 000 were phosphorylatable.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985129 TI - Plasma levels of adenosine 3',5'-monophosphate in the fetoplacental unit and maternal circulation at delivery. AB - Adenosine 3',5'-monophosphate (cAMP) was measured in matched samples of maternal plasma (MV) and umbilical artery (UA) and vein (UV) of 14 normal infants born following an uncomplicated vaginal delivery. There was good correlation between UA and UV cAMP levels, which seems to indicate that the fetoplacental unit maintains a constant equilibrium of the nucleotide concentration in the umbilical circulation. There was a markedly increased UA cAMP concentration at term relative to that of MV, as well as an arteriovenous gradient of plasma cAMP level (UA greater than UV) in the fetoplacental circulation, which strongly suggest that the high cAMP levels in the cord blood originate mainly from the fetus. PMID- 2985131 TI - Lack of efficacy of estrogen supplementation to imipramine in resistant female depressives. PMID- 2985130 TI - Platelet alpha-2-adrenergic dysfunction in negative symptom schizophrenia: a preliminary study. AB - The specific binding to platelet membranes (Bmax) of 3H-clonidine, an alpha-2 agonist, and 3H-yohimbine, an alpha-2 antagonist, was measured in nine drug-free male schizophrenic patients and repeated after 2 weeks of chlorpromazine (CPZ) treatment. Patients with a lower pretreatment Bmax for 3H-clonidine showed a significantly smaller change in Bmax after treatment, less improvement in their clinical state, as indicated by the change in the Global Assessment Scale (GAS), and a lower posttreatment GAS. Also, they had a significantly higher score for negative symptoms on the Affect Rating Scale both before and after treatment. These findings suggest that schizophrenic patients with relatively subsensitive platelet alpha-2-adrenergic receptors, as measured by 3H-clonidine binding, tend to have more negative symptoms and a diminished alpha receptor binding response and diminished clinical response to CPZ. There were no clinical correlations to 3H-yohimbine binding. PMID- 2985132 TI - Nonsuppression of ACTH in lung cancer. PMID- 2985133 TI - D-Ala6-des-Gly10-gonadotropin-releasing hormone ethylamide: absence of binding sites and lack of a direct effect in rabbit corpora lutea. AB - Rat ovarian tissue has been shown to contain high-affinity gonadotropin-releasing hormone (GnRH) receptors, and synthetic GnRH analogues have been shown to inhibit steroid production by rat corpora lutea in vivo and in vitro. These results raise the possibility that an ovarian GnRH-like peptide may be involved in normal luteal regression. We have examined binding of D-Ala6-des-Gly10-GnRH ethylamide (D-Ala) to rabbit corpora lutea, and have investigated the luteolytic activity of this analogue in hypophysectomized, pseudopregnant rabbits. Three hypophysectomized estrogen-treated rabbits were injected with 0.25 mg D-Ala s.c. every 6 h for 48 h during mid-pseudopregnancy, and three were injected with vehicle only. Treatment with D-Ala produced no acute changes in serum progesterone, nor was the time of luteal regression altered. Rabbit anterior pituitary tissue was found to contain high-affinity GnRH receptors (Ka = 7.0 X 10(9) M-1; 188.2 +/- 35.6 fmol/mg protein). However, no similar high-affinity GnRH receptors were detected in rabbit luteal tissue from any stage of pseudopregnancy. Some apparent low-affinity binding was observed, but this displaceable binding was subsequently observed in all control tissues tested. Thus, a potent GnRH analogue does not have any detectable direct effect on steroidogenesis in the rabbit corpus luteum, nor are high-affinity GnRH binding sites present in rabbit luteal tissue. PMID- 2985134 TI - Influence of follicular maturation on luteinizing hormone-, cyclic 3',5' adenosine monophosphate-, forskolin- and cholesterol-stimulated progesterone production in hen granulosa cells. AB - The influence of follicular maturation on progesterone production by collagenase dispersed hen granulosa cells was measured in short-term incubations. Granulosa cells of the largest follicle (F1) produced up to ten times more progesterone than cells from smaller follicles (F3-F5), not only in response to luteinizing hormone (LH), but also when stimulated by exogenous cyclic AMP or forskolin, both of which raise intracellular cyclic AMP levels by nonreceptor-mediated mechanisms. Moreover, when granulosa cell progesterone synthesis was stimulated by incorporating 25-hydroxy-cholesterol into the incubation medium, an identical pattern was obtained. This could be attributed to a corresponding increase in the specific activity of the mitochondrial cholesterol side-chain cleavage enzyme (20,22 desmolase). An increase in the apparent Vmax was observed without a change in the apparent Km values. Pregnenolone substrate at concentrations which raised progesterone production to levels similar to those observed in response to LH stimulation was utilized equally by granulosa cells of mature and developing follicles. However, at high pregnenolone concentrations, granulosa cells of mature follicles converted significantly more of the precursor to progesterone. Assay of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) showed that the enzyme has two Kms: a low Km present in cells of both mature and developing follicles, and a high Km found only in granulosa cells of more mature follicles. It is concluded that LH-promoted progesterone synthesis in granulosa cells of developing chicken follicles is restricted not so much by the availability of receptors and the competence of the adenylate cyclase/cyclic AMP system, but by the activity of key enzymes, principally the cholesterol-20,22 desmolase. PMID- 2985135 TI - Characterization of a seminal plasma-associated inhibitor of human seminal plasma protein kinase. AB - Human sperm-free seminal plasma (HSP) contains inhibitors (I) of the seminal plasma histone kinase activity (HK). One I is dialyzable and the other I is nondialyzable and precipitable by dialysis of HSP against a hypotonic buffer. When the nondialyzable, precipitable I fraction is resolubilized, it inhibits HK in a concentration-dependent manner. Sephadex G-25 column chromatography of whole HSP resolves I in both the void (Vo) and inclusion (Vi) volumes. Rechromatography of the VoI resolves I solely in the Vo. These and other data suggest that the ViI does not originate from the VoI, and that both I activities represent separate molecular entities. VoI was further characterized and found to be heat labile, trypsin and neuraminidase insensitive, and alpha-chymotrypsin sensitive. VoI is not soluble in CHCl3 or CHCl3:CH3OH (2:1) and is not adsorbed by charcoal. Chromatography of VoI on Sephadex G-100 yields a broad peak of I that migrates just past the Vo. VoI has no detectable cyclic AMP (cAMP) binding activity and VoI activity is not affected by coincubation of VoI and HK with cAMP. VoI also does not bind to zinc-chelate or phenothiazine affinity columns. These data suggest that VoI is protein in nature with properties distinct from the class of previously described protein kinase inhibitors. Although the identity of VoI is not known, it does not appear to be the regulatory subunit of a cAMP-dependent protein kinase, calsemin or a zinc binding protein. PMID- 2985136 TI - Trans/13-cis isomerization is essential for both the photocycle and proton pumping of bacteriorhodopsin. AB - We studied an analogue of bacteriorhodopsin whose chromophore is based on all trans retinal. A five-membered ring was built around the 13-14 double bond so as to prohibit trans to 13-cis isomerization. No light-induced photochemical changes were seen, other than those due to a small amount (approximately 5%) of unbleached bacteriorhodopsin remaining in the apomembrane used for regeneration. The techniques used included flash photolysis at room and liquid nitrogen temperatures and Fourier-transform infrared difference spectroscopy. When the trans-fixed pigment was incorporated into phospholipid vesicles, no evidence of light-initiated proton pumping could be found. The results indicate that trans to 13-cis isomerization is essential for the photochemical transformation and function of bacteriorhodopsin. PMID- 2985137 TI - Low density lipoprotein receptor regulation. Kinetic models. AB - The macromolecular species distribution in a receptor-mediated endocytotic pathway was computer simulated based on kinetic data reported in the literature. In the proposed model, the rapidity with which the recycled receptor is shuttled to the cell surface is indicated by the magnitude of k-3, the shuttling constant. The magnitude of k-3 will vary with the experimental conditions, but when this value is large, the internalized receptor is shuttled back to the cell surface with a traverse time of 14 min. Under steady-state conditions, after the cells have been incubated in the presence of LDL for 5 h (M.S. Brown and J.L. Goldstein, Cell 9 (1976) 663), the time required for a receptor to traverse the entire endocytotic pathway is 52 min. Our simulation suggests that normal LDL binding in such a short-term experiment may be independent of receptor synthesis. Thus, the degradation of LDL and resultant build-up of cholesterol would have no apparent inhibitory effect on the down-regulation of receptor synthesis. PMID- 2985138 TI - AMP interaction sites in glycogen phosphorylase b. A thermodynamic analysis. AB - The binding of AMP to activator site N and to inhibitor site I in glycogen phosphorylase b has been characterized by calorimetry, potentiometry and ultracentrifugation in the pH range 6.5-7.5 at 25 degrees C (mu = 0.1). Calorimetric titration data of phosphorylase b with adenosine 5'-phosphoramidate are also reported at pH 6.9 (T = 25 degrees C, mu = 0.1). Calorimetric curves have been analyzed on the basis of potentiometric and sedimentation velocity results to determine thermodynamic quantities for AMP binding to the enzyme. The comparison of calorimetric titration data of AMP and adenosine 5'-phosphoramidate at pH 6.9 supports the hypothesis previously suggested that the dianionic phosphate form of the nucleotide preferentially binds to the allosteric activator site. The thermodynamic parameters for AMP binding to site N are as follows: delta G0 = -22 kJ mol-1, delta H0 = -34 kJ mol-1 and delta S0 = -40 J mol-1 K-1. The binding of the nucleotide to site I was found to be strongly dependent on the pH. This behaviour may be explained in terms of coupled protonations of three groups having pKa values of 6.0, 6.0 and 6.1 in the unbound form and 7.0, 7.5 and 7.2 in the enzyme-nucleotide complex. The thermodynamic parameters for nucleotide binding to site I for the enzymatic form in which all the modified groups are completely deprotonated or protonated have been calculated to be: delta G0 = -7.7 kJ mol-1, delta H0 = -28 kJ mol-1 and delta S0 = -68 J mol-1 K-1 and delta G0 = 28 kJ mol-1, delta H0H = -10 kJ mol-1 and delta S0H = 61 J mol-1 K-1, respectively. These results suggest that attractive dispersion forces are of primary significance for AMP binding to activator site N, although electrostatic interactions act as a stabilizing factor in the nucleotide binding. The protonation states of those residues of which the pKa values are modified by AMP binding to site I highly influence the thermodynamic parameters for the nucleotide binding to this site. PMID- 2985139 TI - Molecular design of an amplification cascade in vision. PMID- 2985140 TI - Biochemical changes induced by Coxsackie B4 virus in short-term culture of mouse pancreatic islets. AB - Isolated mouse pancreatic islets were infected in vitro with two strains of Coxsackie B4 virus--a tissue culture-adapted strain and a mouse pancreas-adapted strain. Within 48 h of infection changes had occurred in the biochemical activities of islets infected with the mouse pancreas-adapted strain of virus. Basal insulin release was increased two-fold in these islets, while glucose induced insulin secretion remained unchanged. Insulin biosynthesis was greatly reduced at a stimulatory concentration of glucose (20 mM), thus leading to a reduced insulin content in these islets. These effects are of importance because they demonstrate that certain strains of Coxsackie B4 virus, like encephalomyocarditis virus, may selectively alter beta-cell function in vitro. PMID- 2985141 TI - Rapid large-scale purification of plasmid DNA by medium or low pressure gel filtration. Application: construction of thermoamplifiable expression vectors. AB - This paper describes a new method of plasmid DNA purification which is fast and reliable enough for most purposes in recombinant DNA technology. The present method does not require the use of toxic chemicals such as phenol or ethidium bromide, costly ultracentrifugation procedures or other processes which can modify the supercoiled structure of the plasmids, such as adsorption on glass fiber. This method is based on the principle of gel filtration chromatography, at low pressure (1 bar) or medium pressure (between 5 and 10 bars), using Sephacryl S1000 or Superose 6B. It permits recovery of plasmids: in preparative quantities (from 300 micrograms to 4 mg), exempt from RNA, DNA and protein contamination, and suitable for various common genetic engineering procedures immediately after purification. To test the reliability of the technique as well as the degree of purification, the plasmids were used to construct thermoamplifiable vectors, carrying the lacUV5 promoter and the 5' end of the beta-galactosidase gene with a single EcoR1 site in each of the three possible translational phases. This set of vectors is designed for the expression of foreign genes as hybrid proteins in Escherichia coli. PMID- 2985142 TI - The in vitro transcription of a rainbow trout (Salmo gairdnerii) protamine gene. II. Controlled mutation of the cap site region. AB - A series of plasmids containing new fusion genes in which the trout protamine gene is placed under the control of the complete herpes virus (HSV-1) tk promoter Pvu II-Bgl II fragment (pM8), or a shortened thymidine kinase (tk) promoter in which the region between the TATA box and the cap site is altered by using the Pvu II-Mlu I fragment (pM7), have been constructed. An additional recombinant plasmid was constructed in which the Bgl II-Ava II fragment of the protamine gene containing the entire protamine promoter but missing the protamine coding region was cloned into pBR322 between the Xho II 1666 and Hind III sites (pP5). For in vitro transcription, a HeLa cell lysate system was prepared and the RNA transcription products, after glyoxalation, were electrophoretically analyzed on 5% polyacrylamide gels. In constructing pM8 the DNA sequence between the tk promoter and the cap site was present while in pM7 it was deleted. Similar multiple transcripts were seen in both cases, indicating that the region between the promoter and the cap site has no effect upon transcription in vitro. The multiple transcripts appear to be due to the presence of a cryptic promoter in the complementary strand of the protamine gene. The activity of this cryptic promoter has been confirmed by comparison of the transcription of plasmid pP5, in which the protamine mRNA coding region has been deleted, with a previously described plasmid, pJBRP (Jankowski JM and Dixon GH (1984) Can. J. Biochem. Cell. Biol. 62, 291-300), containing the intact protamine gene. PMID- 2985143 TI - Molecular cloning of potato spindle tuber viroid (PSTV) cDNA synthesized by enzymatic elongation of PSTV-specific DNA primers: a general strategy for viroid cloning. AB - Different cDNAs were synthesized by primer extension from the RNA of the severe strain KF 440 of potato spindle tuber viroid (PSTV) with the aid of reverse transcriptase using three PSTV-specific DNA molecules as primers. The cDNAs were made double-stranded and cloned into plasmid pBR 322. Various overlapping subgenomic DNA fragments were prepared from these clones and recombined in two different ways. In both cases a PSTV DNA copy was obtained which represented the entire PSTV RNA genome. The sequence of the DNA of one of the resulting full length clones was identical with the original PSTV isolate, whereas the other clone showed one nucleotide change. On the basis of these results the advantages and problems of different strategies for the molecular cloning of the circular viroid RNA genome are discussed. PMID- 2985144 TI - Partial sequence homologies between cytoskeletal proteins, c-myc, Rous sarcoma virus and adenovirus proteins, transducin, and beta- and gamma-crystallins. AB - Computer based sequence comparisons indicate partial sequence homology between human c-myc, Rous sarcoma virus, adenovirus 7, and simian sarcoma virus proteins and the cytoskeletal proteins d9smin, keratin and vimentin. In addition, sections of the oncogene proteins showed partial but significant homology to alpha and gamma subunits of transducin. gamma-II and beta-BP crystallins showed partial but significant homology to the cytoskeletal proteins keratin, vimentin, desmin, alpha and beta-tubulin, and to adenovirus 7 and simian sarcoma virus transforming gene proteins. Beta-BP crystallin showed partial but significant homology to Rous sarcoma virus protein, and to alpha and gamma subunits of transducin. Both crystallins showed partial sequence homology to the GTP-binding protein elongation factor TU from Escherichia coli. These sequence homologies suggest a link between the mechanisms of normal lens cell differentiation, involving modifications to the cytoskeleton and subsequent changes to the pattern of protein synthesis, and mechanisms of neoplastic transformation. Furthermore, the transducin-like region on beta-crystallin may be important for its interaction with lens membranes and the maintenance of short-range order for lens transparency. PMID- 2985145 TI - Effect of the presence of hardened erythrocytes on deformation-orientation characteristics of normal erythrocytes in shear flow studied by the spin label method. AB - The effect of the presence of hardened red blood cells (HRBC) in a mixed suspension on the deformation-orientation characteristics of normal cells in flow is experimentally probed by the electron spin resonance (ESR) spin label method, using a phosphatidylcholine label which does not transfer between the cells. The average deformation-orientation of the normal cells is generally suppressed by the presence of HRBC to different degrees, depending upon the shape and the way the HRBC are prepared. The effects are qualitatively explained by disturbance of laminar shear flow due to the random tumbling of the HRBC. PMID- 2985146 TI - [Role of adrenergic structures in regulating the release by the intestines of hemocoagulating compounds into the blood stream]. AB - Experiments on cats were made to study the capability of adrenaline, tropaphen and propranolol of influencing the intensity of the release of hemocoagulating compounds and anticoagulants from the intestinal vessels and tissues to the bloodstream (perfusate). Adrenaline was found to increase the coagulative activity of the perfusate, provoking an enhanced release into it of thromboplastin, an analogue of plasma factor V and antiheparin compounds and suppressing the release of antithromboplastins. The blockade of the alpha adrenoreceptors was accompanied by a dramatic increase of antithromboplastins to the intestinal perfusate, whereas the depression of the activity of beta adrenergic structures by reduction of the release of tissue thromboplastin inhibitors. It is concluded that regulation of the release of antithromboplastins in the intestine is mediated by the structures similar in their characteristics to alpha- and beta-adrenoreceptors. PMID- 2985147 TI - [Dynamic changes in the response of molluscan neurons to cyclic AMP with repeated injections]. AB - Microiontophoretic injection of cAMP (but not of non-cyclic AMP) into Helix snail neurons induces rapid reversible membrane depolarization. The reversa potential of the cAMP response varies between +10 and -30 mV in different cells. The amplitude of the responses gradually increases (15 experiments), decreases (4 experiments) or remains unchanged (40 experiments) when the interval between the injections is less than 5 minutes. The identified neurons of the different preparations exhibit different types of dynamic changes in the amplitude of the responses to repeated cAMP injections. The possible role of the retaining current in the origin of the potentiation of cAMP responses was investigated. Potentiation of cAMP responses was preserved after the retaining current was switched off only in cases with no membrane depolarization due to spontaneous leakage of the substance from the microelectrode. It is suggested that potentiation or depression of cell responses to repeated cAMP injections is caused by dynamic changes in the cell cAMP system. PMID- 2985148 TI - [Demonstration of individual reactivity in the dynamics of changes in the concentration of mitochondrial cytochromes during the cirrhotic process]. AB - Using repeated operative biopsies the author examined changes in the concentration of cytochromes a, b, c1 and c in mitochondrial protein of hepatocytes during the development of cirrhosis in rabbit liver. Individual responsiveness was discovered at the level of enzymatic constitution of the mitochondria, that is in the shifts in the specific weight of the respiratory chains in the protein of these organelles. PMID- 2985149 TI - [Human erythrocyte prolyl endopeptidase II hydrolysing teprotid, an inhibitor of peptidyl peptidase from snake venom]. AB - The paper is concerned with the action of 1200-fold purified prolylendopeptidase II (PE-E) from human erythrocytes and the action of highly purified prolyl-D-L alanine peptidyl hydrolase (PE-A) from bovine adenohypophysis on teprotide (BPP9a, SQ 20881), a nonapeptide from venom of the snake Bothrops Jararaca--an inhibitor of peptidyl dipeptidase A (carboxycathepsin). Both the purified preparation PE-E and highly purified preparation PE-A split teprotide at the bonds Pro3-Arg4 and Pro5-Gln6. The Pro8-Pro9-OH bond was not split by the two enzymes. The comparative characteristics of the properties of PE-E and PE-A are presented and the possible physiological role of these enzymes is discussed. PMID- 2985150 TI - [Brain blood supply and beta-endorphin and corticotropin content on exposure to thyroliberin]. AB - Thyroliberin produces a marked depressant action on the reflex cerebrovascular constriction reactions. The lack of changes in the content of immunoreactive beta endorphine in blood and CSF indicates that apparently the cerebrovascular effects of the drug are not mediated via the opioid system but are due to a direct influence of thyroliberin on the central mechanisms by which brain circulation is regulated. Thyroliberin increases the blood corticotropin content, which causes an elevation of the arterial blood pressure and cerebrovascular tension. On the contrary, in the CSF the corticotropin level decreases after thyroliberin administration. The data obtained show that there is no correlation between the content of beta-endorphine and corticotropin in blood and CSF under the action of thyroliberin. PMID- 2985151 TI - [Lymphocyte receptors for trophoblastic beta 1-glycoprotein]. AB - The authors have devised a method for rosette-formation to determine TBG receptors on the surface of T lymphocytes. It has been established that among woman peripheral blood lymphocytes, 7.1% of the cells carry TBG receptors, with this percentage increasing maximally up to 50.2% (the first trimester). Among T gamma lymphocytes, 1.2% carry TBG receptors. The amount of such lymphocytes rises 10-fold by the end of pregnancy. PMID- 2985152 TI - [Cyclic AMP content of autonomic ganglia as affected by catecholamines]. AB - The relative content of cAMP was measured in the rat ganglion nodosum, lumbar ganglia of the sympathetic trunk, the main pelvic ganglion and intramural ganglia of the heart. It was observed that the basal level of cAMP in the cardiac ganglia was lower than in other ganglia. The process of stimulation of the cAMP content by noradrenaline was most pronounced in the main pelvic and lumbar ganglia, that by dopamine in the cardiac ganglia. The catecholamines failed to alter the cAMP content in the ganglion nodosum. PMID- 2985153 TI - [Effect of lithium preparations on cardiac arrhythmias due to strophanthin in waking rats]. AB - It was shown in experiments on conscious rats that intravenous injection of strophanthine in toxic doses provokes heart arrhythmias and death of the animals. Lithium drugs (lithium chloride and lithium hydroxybutyrate) injected during arrhythmias led to a short-lived effect of heart rhythm normalization. Lithium hydroxybutyrate was more effective if administered shortly after strophanthine injection, reducing the latter's cardiotoxic effect and preventing the death of the majority of the animals. PMID- 2985154 TI - [Effect of antihypoxic agents on the cyclic nucleotide content in different brain structures in normo-oxia and hypoxia]. AB - A study was made of the effects of isothiobarbamine and guthimine (10 and 50 mg/kg, respectively) on the content of cAMP and cGMP in the brain cortex (BC) and hippocamp under normal conditions and hypoxia. Isothiobarbamine did not change the content of both cyclic nucleotides under normoxia, whereas under hypoxia it reduced the level of the cyclic nucleotides in the BC and raised it in the hippocamp. Guthimine increased their content in the BC and did not change it in the hippocamp under normoxia, whereas under hypoxia it increased the cAMP content in the hippocamp and did not change it in the BC. The cGMP content descended in both the structures under study. PMID- 2985155 TI - [Structural changes in erythrocyte membranes in nephropathy]. AB - Spin probes were used to study alteration of red cell membranes in nephropathy of varying degree of gravity. Iminoxyl radicals of the lipid nature were applied as spin probes, in particular 2-(3-carboxypropyl)-4,4-dimethyl-2-tridecyl-3 oxazolidinyl (probe I). It was shown that in nephropathy, the orderliness parameter increases and the hydrophoby of probe I localized in a red cell suspension of nephropathy patients diminishes as compared with analogous parameters in healthy pregnant women. This attests to both immobilization of the fatty acid chains of phospholipids and to an increase in the polarity of the lipid bilayer in the area of probe localization. It was established that diminution of probe I hydrophoby is in a satisfactory agreement with the disease gravity and the degree of edema in patients. It was noted that alterations discovered in red cell membranes in nephropathy are similar to those seen during activation of lipid peroxidation in membranes. The possibility of lipid peroxidation involvement into the pathogenesis of nephropathy is discussed. PMID- 2985156 TI - [Measurement of the rate of O2 consumption by peritoneal macrophages using electron paramagnetic resonance]. AB - To study the process of activation of macrophages by silicon dioxide particles, use was made of an electrode-free method for measuring the O2 consumption rate. It was discovered that within the first minute of interaction with silicon dioxide particles the rate of O2 consumption by peritoneal macrophages rose 3-4 fold. PMID- 2985157 TI - [Regulation of the cell cycle of B-lymphocytes in mice by substances elevating the levels of intracellular cAMP and cGMP]. AB - Spontaneous velocity sedimentation of B lymphocytes activated by intraperitoneal injection of ovalbumin into mice was used to obtain cell cycle synchronized cells, evidenced by differences in the incorporation of labeled precursors of protein and nucleic synthesis (14C-methionine and 3H-thymidine). The effects of acetylcholine and adrenaline, cAMP and cGMP on the intensity of 3H-thymidine incorporation into mouse B lymphocytes and on the amount of the cells entering mitosis were examined. It was shown that acetylcholine is capable of stimulating whereas adrenaline of inhibitin B lymphocyte entry into the stage of DNA synthesis and egress of these cells from the stage of DNA synthesis to the stage of mitosis. Adrenaline was found to have a reciprocal action. The acetylcholine effect could be mimetized by exogenous cGMP, that of adrenaline by cAMP. Stimulation of the G1/S transition was mediated by intracellular calcium ions but did not depend on exocellular calcium. PMID- 2985158 TI - Reduced peripheral presynaptic adrenoceptor sensitivity following chronic antidepressant treatment in rats. AB - Responses of the isolated vas deferens of the rat to clonidine (inhibition of contractions to field stimulation at 0.1 Hz) and noradrenaline (contraction of longitudinal muscle) were determined after one or 21 daily injections of the animals with desmethylimipramine, clorgyline, selegiline or tranylcypromine. Desmethylimipramine (10 mg kg-1) and clorgyline (2 mg kg-1) increased the clonidine EC50 in the isolated vas deferens after 21 but not after one daily injection(s). Tranylcypromine (5 mg kg-1) increased clonidine EC50 after both one and 21 injections and selegiline (1 mg kg-1) did not affect clonidine EC50 after either one or 21 injections. Only desmethylimipramine had a significant effect on noradrenaline responsiveness, producing an inconsistent decrease in EC50 with a consistent increase in maximum contractile response after 21 but not after one daily injection(s). Clorgyline (10(-5) M) increased the contractile response of the isolated vas deferens to field stimulation and antagonized the inhibitory effect of clonidine when added directly to the isolated tissue preparation. Neither clorgyline (5 mg kg-1), selegiline (1 mg kg-1) nor tranylcypromine (2.5 mg kg-1) affected significantly the inhibitory response to clonidine (1 microgram kg-1) on contractions of in situ sympathetically stimulated vas deferens in pithed rats. These results show that down-regulation of alpha 2-presynaptic adrenoceptors by chronic treatment with desmethylimipramine and clorgyline occurs in peripheral organs as well as in the central nervous system. PMID- 2985159 TI - The adenosine receptor activity of EMD 28422, a purine derivative with reported actions on benzodiazepine receptors. AB - The effects of a novel purine derivative, N6-[2-(4-chlorophenyl)-bicyclo 2.2.2.octyl-(3)]-adenosine (EMD 28422), that has been found to influence central benzodiazepine receptors, has been compared to those of other adenosine analogues such as L-phenylisopropyladenosine (L-PIA), cyclohexyladenosine (CHA) and adenosine-5'-N-ethyl-carboxamide (NECA). EMD 28422 was about 30 times less potent than CHA and 4 times less potent than NECA in displacing bound [3H]-L-PIA from specific binding sites in the rat brain, presumably reflecting adenosine A1 receptors. A similar relative potency was found using depression of field e.p.s.p. in the hippocampal slice in vitro. In isolated fat cells EMD 28422 was antilipolytic, but some 1000 times less potent than L-PIA. In rat isolated hippocampal slices, which have adenosine A2-receptors, EMD 28422 was more than 300 times less potent than NECA and in guinea-pig thymocytes, which similarly have A2-receptors, EMD 28422 was about 60 times less potent. The results are compatible with the opinion that EMD 28422 is a rather weak agonist at adenosine receptors, with limited selectivity for A1- or A2-receptors. The compound is highly lipophilic, which plays a role in determining its potency in a given biological system. The results are discussed in relation to reported adenosine modulation of benzodiazepine receptors. PMID- 2985160 TI - Comparison of the effects of caffeine and procaine on noradrenergic transmission in the guinea-pig mesenteric artery. AB - The effects of caffeine and procaine on noradrenergic transmission in the guinea pig mesenteric artery were investigated by recording electrical responses of smooth muscle cells and by measuring the outflow of noradrenaline (NA) and 3,4 dihydroxyphenylglycol (DOPEG) induced by perivascular nerve stimulation. Caffeine possessed dual actions on the membrane, i.e., at low concentrations (2.5 X 10(-4) 5 X 10(-4)M), it hyperpolarized the membrane and decreased the membrane resistance and at high concentrations (over 2.5 X 10(-3)M) it depolarized the membrane and increased the membrane resistance. Procaine (over 10(-4)M) consistently depolarized the membrane and increased the membrane resistance. The amplitude of the excitatory junction potential (e.j.p.) produced by perivascular nerve stimulation was increased by low concentrations of procaine (2.5 X 10(-5) 10(-4)M) or high concentrations (10(-3)-5 X 10(-3)M) of caffeine and was decreased by low concentrations of caffeine (2.5 X 10(-5)-10(-4)M) or high concentrations of procaine (5 X 10(-4)-10(-3)M). Higher concentrations of caffeine (over 5 X 10(-3)M) induced a spike potential on the e.j.p., while higher concentrations of procaine (over 2.5 X 10(-3)M) inhibited the generation of e.j.ps. Facilitation of e.j.ps produced by repetitive stimulation of perivascular nerves remained unchanged by caffeine, while it was enhanced by procaine at any given concentration (caffeine 2.5 X 10(-4)-10(-3)M; procaine 10(-4)-10(-3)M). The membrane depolarization produced by exogenously applied NA (10(-5)M) was not blocked by pretreatment with procaine. Conduction velocity of perivascular nerve excitation remained unchanged by application of caffeine (up to 5 X 10(-3)M), and was reduced by application of procaine (over 2.5 X 10(-4)M). Outflow of NA during perivascular nerve stimulation remained unchanged by caffeine (10(-4)-3 X 10( 3)M), while it was enhanced by procaine (over 2.5 X 10(-4)M). The outflow of DOPEG was slightly reduced by caffeine (10(-3)-5 X 10(-3)M) and by lower concentrations of procaine (10(-4)-2.5 X 10(-4)M) but was not altered by higher concentrations of procaine (10(-3)-5 X 10(-3)M). It is concluded that in the guinea-pig mesenteric artery, high concentrations of caffeine (over 10(-3)M) increased the e.j.p. amplitude which might be due to an increase in membrane resistance of the smooth muscle cells. No marked effect of caffeine was observed on transmitter release from the nerve terminals. Procaine (over 2.5 X 10(-4)M) increased transmitter release from perivascular nerves and blocked the re-uptake mechanism of released NA. The mechanisms underlying the decrease in ej.p. amplitude by procaine remain to be determined. PMID- 2985161 TI - Sodium load and high affinity ouabain binding in rat and guinea-pig cardiac tissue. AB - An estimation of the actual Na/K-ATPase transport activity in intact cardiac cells was made by measuring the binding of [3H]-ouabain to rat and guinea-pig ventricular strips. At the low [3H]-ouabain concentration of 1 nM equilibrium binding was hardly obtained after an incubation time of five hours. Different procedures known to alter the sodium load of the cardiac preparations influenced [3H]-ouabain binding: the sodium ionophore monensin enhanced [3H]-ouabain binding, the local anaesthetic dibucaine and a reduction of external sodium ion concentration diminished [3H]-ouabain binding; [3H]-ouabain binding was similarly affected by these procedures in the rat and guinea-pig. Since [3H]-ouabain binding occurred predominantly at the high-affinity binding sites of rat myocardium under the applied experimental conditions, it was concluded that these binding sites represent Na/K-ATPase molecules involved in sodium ion transport. PMID- 2985162 TI - Central effects of nicotinamide and inosine which are not mediated through benzodiazepine receptors. AB - The actions of nicotinamide and inosine were investigated on rat cerebellar Purkinje cells using ionophoretic and extracellular recording techniques. Ionophoretic application of nicotinamide or inosine showed that they were potent inhibitors of Purkinje cell firing. This inhibition differed from that induced by benzodiazepines in that it was not reversed by the GABA antagonists bicuculline methiodide and picrotoxin. RO 15-1788, the specific benzodiazepine antagonist, did not reverse the effects of nicotinamide. Chlordiazepoxide has been shown to increase significantly social interaction between pairs of male rats and this increase can be reversed by RO 15-1788, 20 mg kg-1 i.p. Nicotinamide also caused a small increase in social interaction but this effect was not reversed by the benzodiazepine antagonist. Inosine did not increase social interaction. [3H] flunitrazepam binding studies showed that nicotinamide and inosine have only low affinities for the benzodiazepine binding site. These results suggest that while nicotinamide may exert some neuronal depressant and anxiolytic activity, its site of action appears not to be associated with the benzodiazepine receptor site. Similarly, inosine exerts a neuronal depressant effect dissimilar from that of benzodiazepines. PMID- 2985163 TI - In vivo interaction of prostacyclin with an inhibitor of cyclic nucleotide phosphodiesterase, HL 725. AB - Prostacyclin (PGI2) inactivates platelets by stimulation of adenylate cyclase, and its effect can be potentiated in vitro by simultaneous inhibition of cyclic AMP phosphodiesterase. The interaction of synthetic PGI2 and the potent phosphodiesterase inhibitor HL 725 was studied in a model of systemic platelet activation by intravenous injection of collagen. Platelet aggregate formation was evaluated by continuous on-line measurement of the circulating platelet count. Collagen injection in rabbits receiving vehicle caused a 30 +/- 3% decrease in the circulating platelet count. Infusion of PGI2 (0.05, 0.1 and 0.75 micrograms kg-1 min-1) dose-dependently inhibited this decrease. HL 725 (0.5, 1 and 3 micrograms kg-1 min-1) caused a slight but significant effect. Combinations of PGI2 and HL 725, respectively, at 0.25 + 1.0 and 0.1 + 0.5 micrograms kg-1 min-1 inhibited platelet aggregate formation to a greater extent than when either substance was used alone and produced a comparable inhibition to PGI2 at 0.75 micrograms kg-1 min-1. Collagen induced an acute fall in the mean arterial blood pressure (MABP) which also was inhibited by PGI2, HL 725 and their combinations. The infusion of a combination of PGI2 and HL 725 before collagen produced a decrease in the MABP which was greater than when either compound was used on its own. Thus, PGI2 and the phosphodiesterase inhibitor HL 725 interact in vivo to inhibit platelet aggregation and lower MABP. PMID- 2985164 TI - The aftercare of the patient with the neuroleptic malignant syndrome. AB - The neuroleptic malignant syndrome (NMS) is an idiosyncratic reaction to neuroleptic drugs, made up of hyperthermia, muscular rigidity, disturbance of level of consciousness and autonomic dysfunction. It is potentially lethal and should be kept in mind whilst using anti-psychotic drugs; as most patients treated by them require further anti-psychotic treatment, the clinician faces the problem of treating those patients after a NMS episode, yet reports in the literature have generally neglected the problem of late management. A patient suffering from a psychosis and NMS is presented, and a rationale for management offered. PMID- 2985165 TI - Thallium-technetium subtraction scintigraphy as an aid to parathyroid surgery. AB - We describe the application of thallium-technetium subtraction scintigraphy in nine patients with failed previous parathyroid surgery or with tertiary hyperparathyroidism due to chronic renal failure. The technique successfully located all adenomas, but only 45% of hyperplastic glands. The series included three abnormal glands located retrosternally. The technique appeared to be more useful in patients with primary hyperparathyroidism than in tertiary hyperparathyroidism, possibly related to differences in gland mass. We conclude that this method of scintigraphy is a valuable adjunct to the management of patients with parathyroid disorders, particularly those requiring revision surgery. PMID- 2985166 TI - Abdominal pain as a presenting symptom of male germ cell tumour. AB - Twenty-three patients who presented with abdominal pain and were found to have germ cell tumours have been reviewed. The overall survival of these patients was no different from the survival of those presenting with testicular swellings. Intra-abdominal germ cell tumour remains an important differential diagnosis of abdominal pain in men. PMID- 2985167 TI - Malignant fibrous histiocytoma involving the bladder. PMID- 2985168 TI - Chemotherapy for small cell carcinoma. PMID- 2985169 TI - Acute appendicitis and dietary fibre: an alternative hypothesis. AB - This paper examines the time trends in acute appendicitis in Britain in relation to changes in consumption of fibre, meat, and sugar. Knowledge of previous case fatality rates, diagnostic criteria, and management are sufficient to infer general trends in incidence from mortality data. The incidence rose steeply from around 1895 and fell from the 1930s onwards. Contrary to previous statements there is no evidence that disease rates were greatly influenced by the dietary changes in the second world war. It is concluded that dietary changes do not explain the time trends in appendicitis and that the epidemiology of the disease is more readily explained by a primarily infectious aetiology. PMID- 2985170 TI - Prevalence of antibody to human T lymphotropic virus type III by risk group and area, United Kingdom 1978-84. AB - Antibody to human T lymphotropic virus type III (anti-HTLV-III) was sought in 2150 patients in three groups at risk with a radioimmunoassay and an immunofluorescence test. Results by the two methods were closely concordant. Anti HTLV-III was already present in some British homosexuals in 1980 and in some British haemophiliacs in 1981, and since then its prevalence has increased. Of homosexual patients needing laboratory tests for hepatitis B virus infection in 1984, 34% of 282 in London and 5% of 955 in five centres outside London were positive for anti-HTLV-III. Of haemophiliacs sampled in 1984, 38% of 81 were anti HTLV-III positive. Most of the seropositive haemophiliacs were regular recipients of commercial factor VIII concentrates. Few British intravenous drug abusers sampled in 1984 (2.5% of 203) were positive for anti-HTLV-III. These results show that infection with HTLV-III has rapidly become widespread among homosexuals attending sexually transmitted disease clinics and among haemophiliacs receiving pooled blood products. Thus, while many anti-HTLV-III positive individuals may remain asymptomatic, there may soon be a considerable increase in the incidence of the acquired immune deficiency syndrome and related disease in Britain. PMID- 2985171 TI - Racial and other characteristics of human T cell leukemia/lymphoma (HTLV-I) and AIDS (HTLV-III) in Trinidad. AB - Adult T cell leukaemia/lymphoma was first recognised as a clinical entity in southwest Japan. Subsequently the Caribbean has been found to be another area where the disease is endemic, and sporadic cases have been identified in different parts of the world. The human T cell leukaemia/lymphoma virus (HTLV-I) is causally related to adult T cell leukaemia/lymphoma. A subgroup of HTLV, designated HTLV-III, has recently been isolated from many patients with the acquired immunodeficiency syndrome (AIDS) and preAIDS, and there is now evidence that this variant is the primary cause of AIDS. This is the first report from Trinidad to describe 12 cases of adult T cell leukaemia/lymphoma and 14 of AIDS. All were in patients of African descent. No cases were seen in subjects of East Indian descent, who, like those of African descent, comprise as much as 40% of the population. West Indians of African descent may have increased susceptibility to infection with both HTLV-I and HTLV-III. PMID- 2985172 TI - AIDS: the African connection. PMID- 2985173 TI - Electrophysiological study of the corticonuclear projection in the cat cerebellum. AB - Experiments were designed to examine the relationship between the responses of Purkinje cells to natural peripheral stimuli and the location of these neurons within identified zones of the corticonuclear projection in lobule V of the cat cerebellar cortex. It was hypothesized that the corticonuclear zones are sharply demarcated and that the responses of Purkinje cells to a restricted natural stimulus is not localized to only one zone but rather is present and varies in character across these 3 zones. Initially the spatial distribution of the antidromic field potential evoked by stimulating in the fastigial (FN), lateral vestibular (LVN), and anterior interposed nuclei (AIN) was determined in sublobules Va-Vc in unanesthetized decerebrate cats. In some animals the corticonuclear projection was further examined by evaluating the location of Purkinje cells responding antidromically to stimuli in the FN, LVN and AIN, or FN, AIN and the posterior interposed nuclei (PIN). Once a Purkinje cell was identified, its simple and complex spike responses to a step-like flexion extension passive movement of the ipsilateral forepaw were determined. The boundary based on the antidromic activation of Purkinje cells between the fastigial zone (FZ) and the anterior interposed zone (AIZ) in sublobules Va-Vc of the cerebellar cortex was highly reproducible from cat to cat, although there was a slight overlap between these zones based on the antidromic field potential. The FZ-AIZ border was located at 2.1 +/- 0.12 mm lateral and parallel to the midline. The FZ also contained a few cells projecting to the LVN. However, the AIZ only contained neurons projecting to the AIN. The boundary between AIZ and PIZ in lobule Va-Vc was between 3.3 and 3.8 mm from the midline and ran parallel to it. The peristimulus time histograms (PSTHs) of the simple and complex spike activity to a passive forepaw displacement revealed extensive modulation of neurons located across the mediolateral extent of the AI and PIZ. Both the simple and complex spike discharge of Purkinje cells projecting to the FN also were modulated, but to a lesser degree than cells in AIZ or PIZ. The spatial distribution of simple and complex spike responses recorded from Purkinje cells overlapped extensively. The data support previous findings that the corticonuclear projection is organized into longitudinally oriented sagittal zones. Electrophysiologically the boundaries were remarkably reproducible from animal to animal. The results also show that information processing involving the modulation of Purkinje cell activity in response to the forepaw stimulus occurs in all 3 zones examined.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985174 TI - Regional distribution of soluble calcium activated proteinase activity in neonatal and adult rat brain. AB - Calcium dependent proteolytic activity in the soluble fraction of various rat brain regions was assayed using 14C-radiolabelled denatured casein as a substrate. Two forms of activity, distinguishable by their calcium requirement for half maximal activation (5 and 80 microM), were found; both were blocked by sulfhydryl alkylating agents and thiol proteinase inhibitors. Preincubation at 58 degrees C for 10 min also eliminated the high threshold activity. These characteristics are identical to those reported for calcium activated neutral proteinases ('calpains') found in other tissues. Calpain activity varied markedly across brain regions. The greatest values for the high threshold enzyme were found in pons-medulla followed by cerebellum/mesencephalon and finally the telencephalon. The low threshold enzyme had low levels of activity throughout the brainstem and diencephalon and was barely detectable in telencephalic structures. In contrast, a previously described endogenous inhibitor of calpain, 'calpastatin', was found not to vary in its activity across brain regions. Calpain activity was high in the prenatal brain, but while the hindbrain maintained high levels of activity into adulthood, the activity in the forebrain dropped 80% during the early postnatal period. The differences between forebrain and hindbrain activity levels were evident during the first 5 days of the neonatal period, suggesting that glial cell differentiation is not responsible for the regional variations found in the adult. These results are discussed with regard to the possibility that the turnover of anatomical structures differs between brain regions. PMID- 2985175 TI - Ca2+-activated protease activity in frog sciatic nerve: characterization and effect on rapidly transported axonal proteins. AB - Protease activity was studied in the frog sciatic nerve. The activity was measured as the release of TCA-soluble radioactivity from either 3H-labelled proteins transported by rapid axonal transport (AXT) or 3H-labelled ganglionic proteins. In nerve homogenates containing transported substrates, protease activity exhibited two peaks, one around pH 5 and one around pH 8. Ca2+ at 100 microM or higher concentrations only stimulated the latter, which was inhibited by 1 mM parachloromercuric benzoate, a sulphydryl reagent, but unaffected by ATP (1 mM). The proteolytic activity was recovered in the 10(5) g supernatant of the homogenate. In desheathed nerves containing 3H-labelled transported proteins, the protease activity could be activated by exposing the nerve to a Ca2+-ionophore, X 537 A, or to an elevated Ca2+-concentration (50 mM). These conditions were also shown to increase the influx and efflux of 45Ca2+ in the nerves. The results indicate the presence within axons of a Ca2+-activated soluble protease, which degrades rapidly transported proteins. The finding that the protease degraded ganglionic soluble proteins to about the same extent suggests a broad substrate specificity. The present system should be useful for further characterization of protease activity during various physiological conditions. PMID- 2985176 TI - ON and OFF regions in layer IV of striate cortex. AB - In vertical penetrations through the striate cortex in the tree shrew (Tupaia belangeri), we found regions where neural activity was evoked predominantly by light ON. These were followed by regions where responses were evoked predominantly by light OFF. Histological reconstructions indicated that the ON regions were correlated with layer IVa and the OFF regions were correlated with layer IVb. Local application of cobalt chloride produced a transient cessation of visually evoked activity, suggesting that the electrodes sampled cortical activity rather than lateral geniculate nucleus afferents. These data demonstrate that separate ON and OFF regions are present in the tree shrew striate cortex and suggest that spatially separate, parallel ON and OFF afferent channels extend, in this species, at least through the first synapse in the striate cortex. PMID- 2985177 TI - Projections from the functional subdivisions of the frontal eye field to the superior colliculus in the monkey. AB - The frontal eye field (FEF) can be divided into the lateral subdivision representing the central visual field (VF), and the medial subdivision representing the peripheral VF. In this report, we examined neurally projected parts within the superior colliculus (SC) from those electrophysiologically identified subdivisions in the FEF using the horseradish peroxidase anterograde transport method. We found that the lateral subdivision projected into the anterolateral portion of the SC, whereas the medial one into the posteromedial portion, indicating that there is a correlation in the VF representation between the source and the target of this corticotectal system. PMID- 2985178 TI - Projection of phrenic afferents to the external cuneate nucleus in the cat. AB - The present study, performed on anesthetized, spontaneously breathing cats, deals with the projection of group I and II muscle afferents of the phrenic nerve (PN) to the external cuneate nucleus (ECN). Stimulation of the central end of the PN evoked a complex response in the ipsilateral ECN. Two principal components could be distinguished in this potential from the respective absolute refractory periods (ARP) and from the effect of antidromic stimulation in the ECN. Thus, the early group of waves may correspond to recordings of direct fibers and the later group to postsynaptic activations within the ECN. Similar to the forelimb nerves and intercostal nerves of the upper intercostal spaces, the larger muscle afferents of the PN project to the ECN. PMID- 2985179 TI - Electrophysiological studies of a rostral projection from the nucleus raphe magnus to the hypothalamus in the rat and cat. AB - Neurones in nucleus raphe magnus (NRM) and the adjacent reticular formation with rostral projections were identified by their antidromic responses to stimulation at periventricular forebrain sites in rats and cats. In subsequent experiments, the effects of stimulation in the midline of the ventral medulla were tested on the activities of periventricular forebrain neurones. Taken together, the results of these experiments suggest that a direct inhibitory projection may exist from NRM to ventromedial forebrain structures including the anterior hypothalamus/preoptic region in rats in addition to polysynaptic pathways which mediate both excitations and inhibitions in rats and cats. PMID- 2985180 TI - Suprachiasmatic nucleus ablation abolishes circadian rhythms in rat brain neurotransmitter receptors. AB - The effect of suprachiasmatic nucleus (SCN) ablation on rhythms in brain neurotransmitter receptors was studied. In control animals, the rhythms in the number of alpha-adrenergic and benzodiazepine receptors were circadian, whereas the rhythms in beta-adrenergic and acetylcholine receptors were ultradian. SCN ablation resulted in loss of circadian, but not ultradian, rhythms, without change in the 24-h mean numbers. SCN ablation appears to cause a selective loss of circadian function in the regulation of brain receptors. PMID- 2985181 TI - Inhibitory action of adrenocorticotrophin on rat arcuate neurons in vitro. AB - Extracellular recordings were made from 52 units in perifused 300 microns thick slices of rat hypothalamus taken parallel to the ventricular surface of the arcuate nucleus and kept at 37 degrees C in an organ bath. Addition of 8 nM adrenocorticotrophic hormone (ACTH) to the perfusate reversibly and rapidly inhibited 15 of the 38 units found within 300 microns of the ventricular surface (in the arcuate nucleus), and there were no comparable excitations. A prompt and reversible excitation of 6/14 units by ACTH was found when records were made from units found 300-600 microns lateral to the ventricle in the lateral arcuate region. It is suggested that ACTH may be an inhibitory transmitter in the arcuate nucleus and that this action may be an inhibition of ACTH-containing neurons. PMID- 2985182 TI - Comparative localization of neurotensin receptors on nigrostriatal and mesolimbic dopaminergic terminals. AB - Neurotensin (NT), a brain-gut peptide, possesses many biological actions similar to those reported for neuroleptics. Moreover, it has been shown that NT alters dopaminergic activity of both nigrostriatal and mesolimbic pathways. We now report that NT receptors are located on dopaminergic cell bodies in both systems. However, the proportion of NT receptors on presynaptic dopaminergic terminals appears to be different. NT receptor sites are mainly found on presynaptic dopaminergic terminals in the caudate-putamen while they are pre- and post synaptically located in the nucleus accumbens and the olfactory tubercule. NT receptors differential localization in these two pathways could be used as a model to study the comparative physiology of various dopaminergic brain systems. PMID- 2985183 TI - Cerebrospinal fluid levels of angiotensin-converting enzyme in Alzheimer's disease, Parkinson's disease and progressive supranuclear palsy. AB - Angiotensin-converting enzyme (ACE, E.C. 3.4.15.1) has been identified as a normal constituent of human cerebrospinal fluid (CSF). ACE activity in CSF from adult subjects without known neurologic disorder correlated positively (P = 0.002) with age between 50 and 90 years. Patients with moderate degrees of senile dementia of the Alzheimer's type and comparably demented patients with Parkinson's disease or progressive supranuclear palsy exhibited mean levels of ACE activity that were decreased 41, 27 and 53% respectively, compared to the mean level in an age and sex-matched group of neurologically intact individuals. These results raise the possibility that ACE activity in CSF may be an index of neuronal dysfunction in certain central neurodegenerative disorders. PMID- 2985184 TI - Distribution of the pro-opiomelanocortin derived peptides, adrenocorticotrope hormone, alpha-melanocyte-stimulating hormone and beta-endorphin (ACTH, alpha MSH, beta-END) in the rat hypothalamus. AB - Rat hypothalamic nuclei were removed and assayed for adrenocorticotropic hormone (ACTH), beta-endorphin (beta-END) and alpha-melanocyte-stimulating hormone (alpha MSH) content by radioimmunoassay, from the same samples. We also performed immunostaining for these 3 pro-opiomelanocortin (POMC) derived peptides on paraffin embedded serial sections of the hypothalamus. Areas known to project to the external zone of the median eminence receive a dense POMC innervation while those projecting to the posterior pituitary are not innervated. In addition, hypothalamic areas previously suggested to project to medullary autonomic centers are densely innervated. This innervation pattern may provide the morphological basis for the involvement of POMC peptides in neuroendocrine and autonomic functions. The biochemical data raise the possibility that the POMC precursor is processed differently in various brain regions. PMID- 2985185 TI - 4-Aminopyridine leads to restoration of conduction in demyelinated rat sciatic nerve. AB - Whole nerve and single axon recordings were obtained in vitro from rat sciatic nerve demyelinated with lysophosphatidyl choline. Compound action potentials recorded across the lesion site were attenuated in amplitude, and showed conduction slowing and temporal dispersion. Application of the potassium channel blocker 4-aminopyridine (4-AP), markedly enhanced the response recorded through the lesion demonstrating both an increase in amplitude and duration of the response. From single axon recordings, we observed two patterns of change in spike waveform following application of 4-AP: broadening of single axon spikes, and bursting of single axon spikes. Some axons that displayed conduction block along demyelinated segments before 4-AP was applied, showed restoration of conduction. PMID- 2985186 TI - Light evoked release of acetylcholine in response to a single flash: cholinergic amacrine cells receive ON and OFF input. AB - By pharmacologically blocking the inhibitory inputs to the cholinergic amacrine cells of the rabbit retina, we have been able to detect the light-evoked release of ACh in response to a single flash. Under these conditions ACh is released equally at light ON and light OFF. This implies that: the cholinergic amacrine cells receive ON and OFF input; they respond to light with depolarizing transients; and the inputs to this system have a basic symmetry. PMID- 2985188 TI - Estradiol and progesterone influence L-5-hydroxytryptophan-induced myoclonus in male guinea pigs: sex differences in serotonin-steroid interactions. AB - The effects of castration in males and sex differences in the effects of estradiol and progesterone on L-5-hydroxytryptophan (L-5-HTP)-induced myoclonus in guinea pigs were examined. Castration had no effect on L-5-HTP-induced myoclonus in males. There were sex differences in sensitivity to L-5-HTP. In the absence of steroids, L-5-HTP-induced myoclonus was higher in gonadectomized males than females. A low dose of estradiol benzoate (EB; 3.5 micrograms) given 46 h before L-5-HTP (100 mg/kg) enhanced myoclonus in gonadectomized females but not males. However, at a higher dose of EB (10 micrograms) and a lower dose of L-5 HTP (80 mg/kg), myoclonic responding was enhanced in males. These findings indicate that estradiol has a similar effect on L-5-HTP-induced myoclonus in males and females, but do not rule out the possibility of sex differences in sensitivity to L-5-HTP when both sexes are given estradiol priming. When L-5-HTP was given 6 h after 0.5 mg progesterone in estradiol-primed males, myoclonus was enhanced. Progesterone treatment reverses the facilitative effect of EB on L-5 HTP-induced myoclonus in females. Therefore, progesterone has opposite effects on L-5-HTP-induced myoclonus in males and females. These findings were discussed with respect to the interaction of steroids and 5-HT transmission in the regulation of steroid-dependent reproductive behavior. PMID- 2985187 TI - Presynaptic localization of B-50 phosphoprotein: the (ACTH)-sensitive protein kinase substrate involved in rat brain polyphosphoinositide metabolism. AB - This study describes the ultrastructural localization in rat hippocampal tissue in situ and in isolated synaptosomes of the brain-specific phosphoprotein B-50, using affinity purified anti-B-50 immunoglobulins (IgGs). Evidence is presented for the presynaptic localization of B-50 in rat brain. Given this specific localization a model is presented outlining the presumed function of the B-50 protein in the membrane and describing possible neuromodulation by adrenocorticotropin hormone (ACTH)-like peptides. PMID- 2985190 TI - Redox changes in the mouse carotid body during hypoxia. AB - The carotid body of the mouse is ideally suited for in vitro photometric and fluorometric studies using transillumination. It is small, thin and translucent. Photometric recordings were made by using a dual wavelength method. Wavelengths of 550 nm (maximal absorbance, MA) and 540 nm (isosbestic point, IP) were employed to study cytochrome c, 600 nm (MA) and 620 nm (IP) for a pigment presumed to be cytochrome aa3, 580 nm (MA) and 570 or 542 nm (IP) to control for possible effects of hemoglobin (Hb). Exposure of the organ to NaCN (10-50 nM) reduced cytochromes c and putative aa3. Hypoxia (produced by superfusing with a medium equilibrated with 100% N2) reduced cytochrome c but tended to oxidize the presumed cytochrome aa3. These effects were apparent at a medium pO2 of 100 torr or less. Fluorometry revealed reduction of NADH under both cyanide exposure and hypoxia. There was little or no interference by Hb when the preparations were carefully washed of remaining red cells. The precise site of the recorded mitochondria is unknown: they could have been located in the parenchymal cells (type I and/or II), nerve terminals, smooth muscle fibers, etc. Resolution of this point will need using dissociated carotid body cells. Further, the possible presence of a special respiratory pigment responding at a wavelength similar to that exciting cytochrome aa3 has not been discarded. Its study would require isolating carotid body mitochondria. PMID- 2985189 TI - Securinine alkaloids: a new class of GABA receptor antagonist. AB - Experiments were undertaken to determine the site of action of securinine and related convulsant indolizidines. All of these compounds induced tonic seizures in mice, with CD50 values ranging from 11 to 87 mg/kg. The CD50 for bicuculline was found to be 8 mg/kg. Equilibrium binding assays revealed that securinine and dihydrosecurinine inhibit [3H]GABA binding to rat brain membranes with an IC50 of approximately 50 microM, which is some 7 times less potent than bicuculline. Allosecurinine and virosecurinine have IC50 values greater than 1 mM. Both dihydrosecurinine and securinine inhibited GABA-stimulated benzodiazepine binding in rat brain membranes, though they were somewhat weaker than bicuculline in this respect. Other binding assays revealed that securinine and its analogs were inactive as inhibitors of bicuculline-insensitive GABA binding, benzodiazepine, cholinergic muscarinic, and beta-adrenergic receptor binding. In addition, while thiocyanate ion increased the apparent binding potency of bicuculline 10-fold, it had little effect on that of securinine. Extracellular electrophysiological studies on neurons in the cat spinal cord indicated that securinine and dihydrosecurinine blocked the inhibitory action of GABA while having no effect on that of glycine. Allo- and virosecurinine were much less active as GABA receptor antagonists in this test. These results suggest that, like bicuculline, securinine and dihydrosecurinine are selective antagonists of GABA recognition sites on mammalian central neurons. PMID- 2985191 TI - Alpha 1-noradrenergic receptor blockade decreases nuclear estrogen receptor binding in guinea pig hypothalamus and preoptic area. AB - Prazosin, a putative alpha 1-noradrenergic antagonist, reduced nuclear estrogen receptor levels in the hypothalamus and preoptic area of the estrogen-primed guinea pig. Noradrenergic transmission may alter steroid-dependent processes and neuroendocrine function through actions on steroid-concentrating cells. PMID- 2985192 TI - Regulation of biogenic amine methyltransferases by glucocorticoids via S adenosylmethionine and its metabolizing enzymes, methionine adenosyltransferase and S-adenosylhomocysteine hydrolase. AB - This report examines the possibility that glucocorticoids control the degradation of adrenal phenylethanolamine N-methyltransferase and pineal hydroxyindole O methyltransferase by regulating endogenous concentrations of the cosubstrate, S adenosylmethionine, via its metabolic enzymes, methionine adenosyltransferase and S-adenosylhomocysteine hydrolase. Assays for these latter enzymes were established and optimized in the adrenal and pineal glands. The effects of hypophysectomy and dexamethasone or ACTH treatment on these enzymes were monitored along with concomitant changes in methyltransferase activity. Hypophysectomy simultaneously decreases methionine adenosyltransferase and S adenosylhomocysteine hydrolase activity in both tissues. Dexamethasone administration to hypophysectomized animals does not alter either methionine adenosyltransferase activity or S-adenosylhomocysteine hydrolase activity in the adrenal gland. However, it does increase S-adenosylhomocysteine hydrolase activity in the pineal gland. In contrast, ACTH administration restores both enzymes in the adrenal while being ineffective in the pineal. These results suggest that glucocorticoids may be regulating S-adenosylmethionine levels and methyltransferase activity via the metabolic enzymes, methionine adenosyltransferase and S-adenosylhomocysteine hydrolase, but that glucocorticoid control may be both tissue- and drug-specific. PMID- 2985193 TI - Autoradiographic localization of calcium channel antagonist receptors in rat brain with [3H]nitrendipine. AB - In vitro autoradiographic techniques have been used to localize [3H]nitrendipine binding sites in the rat brain. The superficial cerebral cortex, the ventral, lateral and posterior nuclei of the thalamus, the molecular layer of the dentate gyrus, the substantia nigra and the external plexiform layer of the olfactory bulb, all contain high densities of silver grains. The level of binding sites are greatly reduced in areas low in synaptic connections. The corpus callosum, the fimbria, the alveus hippocampi and the dorsal commissure of the fornix all lack specific silver grains, as does the lateral olfactory tract of the olfactory bulb. Specific silver grains are not found in the habenula or the hypothalamus. Grains are not associated with blood vessel profiles. The discrete localizations of [3H]nitrendipine binding sites suggest a specific synaptic role. PMID- 2985195 TI - The regional distribution of receptors for vasoactive intestinal polypeptide (VIP) in the rat central nervous system. AB - The regional distribution of receptors for vasoactive intestinal polypeptide (VIP) was studied in the rat central nervous system (CNS). The specific binding was highest in cerebral cortex, limbic forebrain and cerebellum, whereas moderate to low binding was found in hypothalamus, thalamus, brainstem and pituitary. The lowest binding was observed in pons and spinal cord. Scatchard analysis showed curvilinear plots with upward concavity, which was interpreted as two classes of binding sites. The Kd values were similar in all regions and calculated as 2.4 and 62 nmol/liter, respectively. The variations of specific [125I]VIP binding were due to differences in the total amount of receptors and were in the range of 1.7-8.6 pmol per mg protein. The regional distribution of VIP receptors was parallel with the occurrence of VIP-containing nerve terminals with exceptions of cerebellum, olfactory areas and nucleus caudatus, where a greater number of receptors than expected from the VIP content was found. In these regions, VIP may interact with receptors for a different, but homologous neuropeptide. In conclusion, the regional distribution of VIP receptors in CNS gives further evidence for the role of VIP as a central neurotransmitter. PMID- 2985194 TI - Effects of kynurenate on root potentials evoked by synaptic activity and amino acids in the frog spinal cord. AB - The effects of kynurenate (Kyn) on synaptic- and excitatory amino acid-mediated responses in isolated, hemisected spinal cords of frog were examined. Kyn (0.5 mM) rapidly and reversibly blocked greater than 90% of the synaptically mediated ventral root potential (VRP) produced by stimulation of the dorsal root. Spontaneous activities recorded from both ventral and dorsal roots were also reversibly blocked by Kyn. However, Kyn had no effect on action potentials or excitability per se, nor was it a general inhibitor of synaptic transmission since Kyn concentrations as high as 2.5 mM had no effect on synaptically mediated dorsal root potentials produced by stimulation of the ventral root. In addition, Kyn had no effect on synaptic transmission in sympathetic ganglia of frog. Although Kyn (2.5 mM) by itself produced no ventral root response in spinal cords treated with tetrodotoxin, it antagonized those induced by the excitatory amino acids N-methyl-D,L-aspartate, quisqualate, kainate, aspartate, and glutamate. The ventral root responses to all concentrations of quisqualate tested were depressed by 2.5 mM Kyn. In addition, when Kyn was washed out, the rate of recovery from Kyn block was accelerated by the presence of quisqualate. These results indicate that quisqualate and Kyn compete for common binding sites. However, low concentrations of Kyn (e.g. 0.1 mM) potentiated the peak of the response to saturating concentrations of quisqualate by as much as 30%. The durations of the potentiated quisqualate responses were significantly shorter than the control responses. Thus, Kyn does not act simply as a competitive inhibitor of quisqualate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985196 TI - gamma-Aminobutyric acid-mediated inhibition at cholinergic synapses formed by cultured retinal neurons. AB - The purpose of this study was to investigate the effect of gamma-aminobutyric acid (GABA) on the function of synapses formed by cholinergic neurons derived from the chick retina. We used an experimental culture system in which striated muscle cells served as postsynaptic targets for cholinergic neurons of the retina. This cell culture system permitted the physiological monitoring of acetylcholine release at synapses formed by retinal neurons. By plating a low density of dissociated retinal cells with myotubes, it was possible to study relatively isolated, presynaptic cholinergic neurons. We found that GABA and its agonists, muscimol and isoguvacine, inhibited spontaneous transmission at retina muscle synapses. These inhibitory effects were reversibly blocked by bicuculline, a GABA receptor antagonist. The benzodiazepine, flurazepam, potentiated GABA mediated inhibition. Overall, our findings suggest a direct inhibitory action of GABA on the cholinergic retinal neurons studied in our cell culture system. PMID- 2985197 TI - The relationship between cross-correlation measures and underlying synaptic events. AB - The relationship between an excitatory postsynaptic potential (EPSP) and the cross-correlation derived from the spike train evoked by a Poisson stimulus train was studied by computer stimulation. The neuron model had a constant threshold and an absolute refractory period. Following a spike, the membrane potential was determined by ongoing activity, i.e., it was not 'reset'. Under all conditions studied, the rise time of the cross-correlation peak accurately reflected EPSP rise time. The agreement between the half-width of the cross-correlation peak and that of the underlying EPSP depended upon the EPSP amplitude, the rate of the driving Poisson input, and the time course of the EPSP. Over a broad range of experimentally relevant conditions, the half-width of the cross-correlation peak deviated from that of the underlying EPSP by less than 30% and bore little resemblance to that of the EPSP's time derivative. PMID- 2985198 TI - An enkephalinergic projection from the hypothalamic paraventricular nucleus to the hypothalamic ventromedial nucleus of the rat: an experimental immunohistochemical study. AB - The distribution and origins of enkephalin-like immunoreactive (ENK-IR) fibers in the nucleus ventromedialis hypothalami (vm) of the rat were examined using immunohistochemistry. A dense plexus was evenly distributed in the vm with no regional differences. A group of ENK-IR neurons was concentrated in the ventrolateral portion of the vm. The destruction of the hypothalamic paraventricular nucleus (pv) which contained numerous ENK-IR neurons, resulted in a marked decrease in ENK-IR fibers in the vm on the operated side. In addition, the destruction of the magnocellular portion of the pv, while leaving most of parvocellular portion intact, failed to substantially decrease the ENK-IR fibers in the vm, suggesting that these fibers originate from ENK-IR neurons located in the parvocellular portion of the pv. The present study further showed that the axons of these neurons first proceeded laterally to the perifornical area, next ran ventrolaterally to the ventrolateral portion of the anterior hypothalamic nucleus and finally turned medially to the vm. PMID- 2985199 TI - Circadian variations in radioligand binding to muscarine receptors in rat brain dependent upon endogenous agonist occupation. AB - Although total levels of muscarine receptors in the rat forebrain and brainstem are constant at different times of day, there are large circadian changes in the numbers and carbachol-affinities of unoccupied receptors, which appear to reflect varying levels of endogenous acetylcholine-receptor complexes and the activity of cholinergic synapses. PMID- 2985200 TI - Inhibitory influence of GABA on central serotonergic transmission. Involvement of the habenulo-raphe pathways in the GABAergic inhibition of ascending cerebral serotonergic neurons. AB - In order to explore the anatomical nature of the inhibitory GABAergic control of cerebral serotonergic neurons exerted at the level of the anterior raphe cells in the rat, we have studied the effect of GABA agonist agents (given systemically or infused locally into the raphe dorsalis or medianus) on cerebral 5 hydroxytryptophan (5-HTP) accumulation after lesion or pharmacological manipulation of various raphe inputs. Destruction of noradrenergic pathways by local injection of 6-hydroxydopamine in the pedunculus cerebellaris superior or by systemic injection of DSP-4 (50 mg/kg i.p.), or alteration of central dopaminergic transmission (by systemic administration of apomorphine or haloperidol) failed to modify the ability of progabide (400 mg/kg i.p.) or dipropylacetamide (150 mg/kg i.p.) to diminish 5-HTP accumulation in the striatum, hippocampus and substantia nigra. In contrast, electrolytic lesion of the habenular nuclei blocked the ability of these compounds (given systemically) to reduce 5-HTP accumulation both in serotonergic nerve terminal and cell body (raphe dorsalis and medianus) areas. A similar blockade of the effects of GABA mimetics was seen after ibotenate-induced lesion of the habenula but not after electrolytic lesion of the stria medullaris (which conveys most of the afferents to the habenula). Acute cessation of impulse flow in the habenulo-raphe tract also prevented the depamide-induced diminution of cerebral 5-HTP accumulation. Finally, interruption of nerve transmission in the habenulo-raphe pathways (by means of electrolytic lesion of the habenula or fasciculus retroflexus) blocked the ability of GABA (100 micrograms) or muscimol (50 ng) injected into the raphe dorsalis or medianus to reduce 5-HTP accumulation in the corresponding serotonergic nerve terminal areas. It is concluded that the GABAergic inhibition of ascending serotonergic neurons exerted in the raphe dorsalis and medianus depends upon an ongoing neuronal activity in the habenulo-raphe pathways; GABA may exert its inhibitory control over serotonergic neurons by tuning down a facilitatory influence on these cells exerted by the habenula. PMID- 2985201 TI - Inhibitory influence of GABA on central serotonergic transmission. Raphe nuclei as the neuroanatomical site of the GABAergic inhibition of cerebral serotonergic neurons. AB - Acute injection of the gamma-aminobutyric acid (GABA) mimetics progabide, aminooxyacetic acid, gamma-acetylenic GABA and dipropylacetamide reduced 5 hydroxytryptophan (5-HTP) accumulation in serotonergic nerve terminal regions (prefrontal cortex, olfactory tubercle, septum, striatum, hypothalamus, hippocampus, substantia nigra, cerebellum and spinal cord) as well as in corresponding cell body areas (raphe dorsalis, medianus, pontis and magnus). This effect was antagonized by bicuculline. The inhibition of serotonin (5-HT) synthesis induced by a single progabide administration was accentuated on repeated treatment in the striatum, prefrontal cortex and cerebellum but was similar to that seen after acute treatment in the other areas. Local infusion of high concentrations of GABA or GABA mimetics into the striatum, septum or substantia nigra failed to modify 5-HTP accumulation in these areas. Cerebral hemitransection antagonized the ability of progabide (1200 mg/kg i.p.) to diminish 5-HTP accumulation in the striatum, hippocampus and prefrontal cortex. Intra-raphe dorsalis infusion of muscimol (0.1-100 ng) or GABA (1-100 micrograms) decreased 5-HT synthesis in the corresponding projection areas (e.g. striatum, substantia nigra, cortex) but not in the hippocampus or cerebellum. Conversely, intra-raphe medianus infusion of these drugs diminished 5-HTP accumulation in the corresponding projection areas (e.g. hippocampus, septum, cortex) but not in the striatum or cerebellum. Intra-raphe dorsalis or medianus injection of GABA antagonists (bicuculline, picrotoxinin, RU-5135) was without effect on cerebral 5 HT synthesis but antagonized the diminution of the amine synthesis observed in corresponding projection areas after intra-raphe dorsalis or medianus infusion of muscimol or GABA. These results suggest that GABA exerts an inhibitory (non tonic) control over central serotonergic neurons which is mediated via GABA receptors located in the raphe nuclei. PMID- 2985202 TI - Differential modulation of gamma-aminobutyric acid receptors by caprolactam derivatives with central nervous system depressant or convulsant activity. AB - The effects of a series of caprolactam derivatives with central depressant, convulsant or muscle relaxant activity were investigated upon gamma-aminobutyric acid (GABA) receptor-ionophore binding to rat brain membranes using [3H]GABA, [3H]GABA, [3H]muscimol and [35S]-tert.-butylbicyclophophorothionate ([35S]TBPS) as ligands, and GABA responses in mouse spinal cord neurones in dissociated cell culture. Some caprolactams produced a picrotoxin-like chloride-dependent partial inhibition of muscimol binding and were potent inhibitors of TBPS binding. One compound that was further investigated (4,4,6,6-tetramethylhexahydro-2H-azepin-2 one), inhibited GABA responses and increased the frequency of paroxysmal depolarizations in cultured neurones. Other caprolactams enhanced muscimol binding and were relatively weak inhibitors of TBPS binding, and one (3,3-diallyl 6,6-dimethylhexahydro-2H-azepin-2,4-dione) was shown to enhance GABA responses and produced quiescence of activity in cultured neurones. There was a direct correlation between caprolactam effects on muscimol binding in the presence of chloride ions and their effects on TBPS binding suggesting a similar site of action for the caprolactams influencing the binding of these two ligands. For the two classes of caprolactams, with respect to inhibition or enhancement of muscimol binding, there appeared to be a relationship between in vitro effects and their convulsant or depressant activity in mice. Caprolactams may be useful low molecular weight probes for the study of GABA receptor-ionophore complexes. PMID- 2985203 TI - Postsynaptic firing during repetitive stimulation is required for long-term potentiation in hippocampus. AB - Long-term potentiation (LTP) in the hippocampus is a long lasting enhancement of the postsynaptic evoked response following high frequency, repetitive stimulation of afferents. The extracellularly recorded action potential (population spike) can be reversibly blocked, without affecting the extracellularly recorded excitatory postsynaptic potential, by focal application of gamma-aminobutyric acid, tetrodotoxin, or pentobarbital, to the CA1 pyramidal cells of the hippocampal slice. When the population spike is blocked during repetitive stimulation, LTP does not occur. It appears that postsynaptic firing of action potentials during repetitive stimulation is necessary to produce LTP. PMID- 2985204 TI - An analysis of the increase in granule cell excitability accompanying habituation in the dentate gyrus of the anesthetized rat. AB - Repeated low-frequency stimulation of the perforant path results in a decrement in the population EPSP and population spike recorded in the hilus of the dentate gyrus. The EPSP decrement is accompanied, however, by an increase in the population spike height/population EPSP slope relation, suggesting that an increase in granule cell excitability also occurs. The present experiments explored the mechanisms of this apparent increase in excitability using standard field potential recording techniques to assess perforant path input/output curves in rats anesthetized with sodium pentobarbital. Low-frequency homosynaptic stimulation (512 pulses, 1 Hz) of the perforant path resulted in a decreased spike threshold and overall shift to the left of the function relating population spike height to EPSP slope. These changes were consistently produced, even when granule cell discharge was inhibited by conditioning stimulation of the contralateral hilus. On the other hand, low-frequency heterosynaptic (lateral perforant path) or antidromic (mossy fiber) driving of the granule cells only slightly increased the medial path spike/EPSP relation, and did not alter the spike threshold. The excitability shift accompanying habituation was qualitatively different from that associated with long-term potentiation, but these shifts did not summate. The interpretation which best explains these various results is that granule cell excitability is increased during low frequency perforant path stimulation by a process of disinhibition, caused by habituation of perforant path excitatory synaptic drive onto feed-forward inhibitory interneurons. PMID- 2985205 TI - Rapid swelling of neurons during synaptic transmission in the bullfrog sympathetic ganglion. AB - Neurons in the lumbar sympathetic ganglion of the bullfrog were found to respond to presynaptic stimulation with rapid mechanical changes. These changes represent swelling of the presynaptic nerve terminals, followed by swelling of the somas of the postsynaptic neurons. After complete blockade of impulse transmission with D tubocurarine, swelling of the neurons associated with the excitatory postsynaptic potential is observed. PMID- 2985206 TI - Early adrenalectomy increases myelin content of the rat brain. AB - Rats were adrenalectomized (ADX) or sham-operated (SHAM) on the 11th day of life and killed on days 35-36, 63, or 151-153 for the isolation of cerebral myelin from each animal. Despite having lower overall body weights, ADX rats had heavier cerebra than SHAM control rats at all ages. Mean cerebral weight increases were 10.0% at day 35-36, 15.3% at day 63, and 16.7% at day 151-153. Recovered myelin dry weights were even more elevated in the ADX rats, but only at day 63 (41.7% increase) and 151-153 (42.1% increase). At both of these ages, there was a clear linear relationship between cerebral wet weights and the amount of myelin recovered from the cerebra. Analysis of the day-63 myelin samples showed no group differences in total cholesterol or protein concentration or in the specific activity of the myelin marker enzyme 2':3'-cyclic-nucleotide 3'-phosphodiesterase (CNP). However, myelin isolated from the ADX rats appeared to be deficient in both galactolipid and phospholipid. Optic nerve myelination was assessed in all animals by measuring CNP activity in homogenates prepared from this tissue. No difference between ADX and SHAM rats was observed at any age. These results indicate that early adrenalectomy stimulates myelin deposition in the rat brain as part of a more general, long-lasting enhancement of brain growth. Myelin from the brains of ADX animals may be slightly abnormal in its lipid composition. Finally, the optic nerve data may mean that myelination is not affected equally in all areas of the CNS by the loss of adrenal glands. PMID- 2985207 TI - Ontogenetic development of serotoninergic neurons in the brain of a teleost, the three-spined stickleback. An immunohistochemical analysis. AB - The ontogenetic development of serotoninergic neurons in the brain of the stickleback was investigated with the indirect immunocytochemical peroxidase antiperoxidase technique, using a specific antibody to serotonin (5 hydroxytryptamine, 5-HT). Formation of neuronal populations takes place during embryonic development. By 80 h after fertilization, the first 5-HT perikarya have appeared in the ventricular zone of the hypothalamus (nucleus recessus lateralis) and the raphe region. At 108 h the first 5-HT perikarya can be observed in area praetectalis. At 118 h a transient group of 5-HT neurons appears rostral to the nucleus recessus lateralis, and at this same age the first 5-HT perikarya may be visualized in nucleus recessus posterioris. A group of 5-HT neurons appears in the dorsolateral tegmentum at 166 h (one day after hatching, which occurs at 120 144 h after fertilization). Differentiation of the neuronal populations, in terms of migration and formation of subdivisions, starts between 80 h and 94 h, and seems to be completed between 1 and 5 days after hatching. Raphe nuclei form an anterior group comprising nuclei raphe dorsalis, raphe medialis and a ventrolateral group, and a posterior group comprising a nucleus raphe pallidus/obscurus complex, a lateral nucleus reticularis paragigantocellularis and a ventromedial nucleus raphe magnus. The posterior and ventral raphe nuclei, which are well developed at the time of hatching, have not been visualized in the adult stickleback. While formation of 5-HT neuronal systems, as well as their primary efferent pathways, takes place during early ontogenetic development, the establishment of terminal areas and their subsequent differentiation apparently takes place during later ontogenetic stages. Most presumptive target areas are penetrated by 5-HT axons at hatching, although terminal formation does not seem to start until later. A considerable number of 5-HT neuronal groups present in the embryonic and newly hatched stickleback have not been visualized in the adult stickleback. This may be due to selective cell death, changes in transmitter phenotype or maturation of axonal transport processes during development. PMID- 2985208 TI - Neonatal 6-hydroxydopamine lesion of spinal norepinephrine terminals fails to affect the ontogeny of swimming behavior. AB - Newborn rats were injected either systemically or intraspinally with 6 hydroxydopamine. Both treatments drastically reduced spinal norepinephrine (NE), while the former also reduced forebrain NE. Spinal NE loss did not affect the ontogeny or the adult expression of swimming, a sensitive test of complex patterned locomotion. Spinal NE is not essential to either the ontogeny of complex motor behavior or to the generation of such behavior in adult rats. PMID- 2985209 TI - Developmental changes in the passive membrane properties of an identified molluscan neuron. AB - We have studied developmentally the electrotonic fate of synaptic potentials in an identified giant neuron of the Pulmonate mollusc Achatina fulica. Our experimental preparation enabled us to sample spontaneously occurring excitatory postsynaptic potentials (EPSPs) from embryogenesis onward, while concomitantly analyzing the postsynaptic membrane properties. We observe non-monotonic changes in the electrotonic attenuation of synaptic inputs which invade the somata. Our data suggest that non-monotonic changes in the specific membrane resistance can account for these observations. PMID- 2985210 TI - Effect of delta 9-THC on brain and plasma catecholamine levels as measured by HPLC. AB - The effects of chronic administration of delta 9-tetrahydrocannabinol (delta 9 THC) on the plasma and brain catecholamine (CA) levels were measured using high performance liquid chromatography-electrochemical detection (LC-EC) system. Intact male rats were injected daily with vehicle (50 microliter oil) or with delta 9-THC (3 mg/kg body wt) over a period of 25 days. Trunk plasma and tissue from preoptic area (POA) and mediobasal hypothalamus (MBH) were collected and catecholamine levels were detected by LC-EC system coupled to an electronic integrator. Alumina extract of tissue and plasma samples, spiked with the internal standard (dihydroxybenzylamine), were injected into the LC-EC system; the CA were chromatographed and eluted within 12 minutes using sodium phosphate buffer as the mobile phase. delta 9-THC treatment resulted in a significant decrease in plasma and MBH levels of norepinephrine (NE), epinephrine (E), POA levels of NE; and significant increases in MBH levels of dopamine (DA) and dihydroxyphenylacetic acid (DOPAC). Our study indicates for the first time that delta 9-THC treatment significantly alters not only the POA and MBH CA levels, but also the plasma CA levels. PMID- 2985211 TI - [The effect of forskolin and triiodothyronine on the production of cyclic adenosine monophosphate in the thymocytes of rats]. PMID- 2985212 TI - Complexes of cytochrome caa3 from the thermophilic bacterium PS3 formed with ligands and during catalytic activity. AB - PS3 (thermophilic bacterium) cytochrome caa3 reacts slowly with cyanide which forms a low-spin complex with the CuBa3 centre. Partial reduction under catalytic conditions increases the rate of cyanide binding, and the reaction constant is rather similar to that of the mammalian enzyme, but the partially reduced complex dissociates more rapidly than does the corresponding eukaryotic complex. A simple biphasic reaction can account for the results obtained. The azide complex of partially reduced PS3 cytochrome caa3 shows an alpha-peak blue shift similar to that of the mammalian enzyme. PS3 cytochrome caa3 forms an oxyferri ("oxygenated") species like the mammalian enzyme, but does not undergo high- to low-spin changes during the aerobic steady state with ascorbate and N,N,N',N' tetramethyl-p-phenylenediamine as substrates. Interactions seen in cytochrome oxidase-ligand reactions and its spin-state changes are therefore intrinsic to the enzyme's large catalytic subunits and do not require the presence of the small nuclear-encoded subunits found in eukaryotic systems. PMID- 2985213 TI - A field trial to evaluate the efficacy of a combined rotavirus coronavirus/Escherichia coli vaccine in dairy cattle. AB - A field trial was designed to determine the efficacy of a combination rotavirus coronavirus/Escherichia coli vaccine on dairy farms in southwestern Ontario. In Part A of the trial, 321 cows on 15 farms were randomly assigned to either vaccination or placebo groups. On eight farms, 50% of the dams were vaccinated, while on the other seven farms, 80% of the dams were vaccinated. In Part B of the trial, 26 farms were randomly assigned to either a total vaccination program or to no vaccination program. Mortality, disease occurrence and weight gains were recorded on all calves for the first two weeks of life. In Part A, 23.5% of all calves were treated in the first two weeks of life, 20.9% were treated specifically for scours and 3.6% of live-born calves died. Enteropathogenic E. coli was identified on 13 of the 15 farms, rotavirus on 11 and coronavirus on ten. At least one of the three potential pathogens was found on every farm. There were no significant differences between calves from placebo-treated and vaccine treated dams with regard to the proportion treated for all diseases, or for scours, or the proportion which died. Neither were there differences in days to first treatment for all diseases (seven days on average), days to first scour (6.7 days), duration of treatments (3.9 days for all diseases, 3.7 days for scours), or estimated weight gains (0.5 kg/day to 14 days). These results were not altered when the presence or absence of enteropathogenic E. coli, rotavirus or coronavirus on the premises was accounted for.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985214 TI - Effect of maternal antibody upon vaccination with infectious bovine rhinotracheitis and bovine virus diarrhea vaccines. AB - This report presents the normal rate of decay of maternal antibody and the influence of maternal antibody on responses to a single vaccination with modified live bovine virus diarrhea and infectious bovine rhinotracheitis virus vaccines at 196 days of age and on response to vaccinations with the same vaccines given twice at 84 and 196 days of age. Passive immunity decreased to near zero over the first six months of life for both bovine virus diarrhea and infectious bovine rhinotracheitis controls. All calves seroconverted to bovine virus diarrhea vaccine at 84 days of age, even though high levels (greater than 1:32) of maternal antibodies were present. These calves did not seroconvert to infectious bovine rhinotracheitis vaccine at 84 days of age when high levels (less than 1:16) of maternal antibodies were present. Calves responded well to bovine virus diarrhea and infectious bovine rhinotracheitis vaccines given only once at 196 days of age after passive immunity disappeared. Calves which were revaccinated with infectious bovine rhinotracheitis seroconverted showing a more rapid response than the single vaccinates. Those revaccinated with bovine virus diarrhea showed an immediate response of small magnitude. PMID- 2985215 TI - Equine parvovirus: initial isolation and partial characterization. AB - A viral agent was isolated from the fetal liver of an aborted equine fetus. The isolate hemagglutinated red blood cells from guinea pig, rhesus monkey and rooster. By hemagglutination inhibition tests, the isolate was shown to be antigenically distinct from parvoviruses of bovine and canine origin. Specific hemagglutination inhibiting antibody against the viral isolate was exhibited by 26 of 136 horse sera tested. The isolated virus showed properties compatible with those of an autonomous parvovirus including size, morphology, stability to ether treatment and heating to 56 degrees C, the presence of a 5300 base DNA genome, characteristic protein composition and density (1.405 g/mL). The virus was classified as an equine parvovirus. PMID- 2985216 TI - Protection from parainfluenza-3 virus and persistence of infectious bovine rhinotracheitis virus in sheep vaccinated with a modified live IBR-PI-3 vaccine. AB - Ewes (N = 7) and their lambs (N = 12) were vaccinated with a commercial modified live infectious bovine rhinotracheitis-parainfluenza type 3 virus vaccine. Both the vaccinated ewes and lambs and a group of unvaccinated ewes (N = 8) and their lambs (N = 13) were subsequently challenged with virulent parainfluenza type 3 virus. Although absolute immunity to infection and clinical response was not conferred, the clinical response was less severe in vaccinated lambs. Vaccinated animals also shed parainfluenza type 3 virus in nasal secretions for a shorter time than nonvaccinated animals. Some vaccinated lambs developed a persistent infectious bovine rhinotracheitis virus infection that was recrudesced by treatment with dexamethasone. It was concluded that vaccination was of benefit in reducing the severity of infection with parainfluenza type 3 virus. However, the inclusion of infectious bovine rhinotracheitis virus in a vaccine for sheep respiratory tract disease is highly questionable as it might increase the risk factor associated with vaccination. The consequences of the persistence of infectious bovine rhinotracheitis virus are now known. PMID- 2985217 TI - Adjuvant enhancement of humoral immune response to chemically inactivated bovine viral diarrhea virus. AB - Potentiation of the antibody response to inactivated bovine viral diarrhea virus by immunological adjuvants was studied in guinea pigs and cattle. The inactivated bovine viral diarrhea virus alone was demonstrated to be a weak immunogen. Addition of either 2 mg per mL diethylaminoethyl-dextran or 5% alhydrogel to inactivated bovine viral diarrhea virus did not or only slightly stimulated the antibody response; the combined adjuvants induced a significantly higher titer. A higher concentration (20 mg per mL) of diethylaminoethyl-dextran, on the contrary, suppressed the immune potentiation by the dual adjuvants. Combination of Bordetella bronchiseptica and alhydrogel adjuvants stimulated a high titer of antibody. The titer was further elevated upon revaccination and was significantly higher than that induced by alhydrogel alone. In cattle, alhydrogel enhanced the immune response and the additional inclusion of diethylaminoethyl-dextran did not cause a significant potentiation of the immunity. However, the antibody decay rate was significantly slower when stimulated by the combined adjuvants. PMID- 2985218 TI - Metabolic and cyclic nucleotide enzyme activities in muscle and nonmuscle cells of rat heart during perinatal development. AB - Enzyme activities related to aerobic metabolism and cyclic nucleotides were evaluated in muscle and nonmuscle cells of rat heart. The perinatal period from weaning to adult was studied. Malate dehydrogenase, citrate synthase, and 3 hydroxyacyl-CoA dehydrogenase activities of nonmuscle cells equal or exceed muscle cell activities in the weanling heart. Aerobic enzymes remain unchanged in nonmuscle cells during growth; however, muscle cell activities are enhanced. Adenylate cyclase and guanylate cyclase activities are higher in heart homogenates of weanling than adult rats. Despite elevated adenylate cyclase activity, cyclic AMP levels are identical in weanling and adult rats. Cyclic GMP levels are twofold higher in weanling than in adult rats. Muscle cell metabolism and cyclic nucleotide levels are associated with growth-related changes in heart function and cellularity, respectively. PMID- 2985219 TI - Effect of weaning on the activities of glycogen synthase and phosphorylase in rat liver. AB - The effects of weaning on the level of glycogen and the activities of glycogen synthase and phosphorylase were determined in rat liver. Glycogen levels in rat liver increased at the start of the weaning period and reached a plateau on postnatal day 20. The active form of glycogen synthase increased until postnatal day 19 and then declined. Total glycogen synthase (active + inactive) remained high during the suckling period and declined to a new low level during the weaning period. The activity ratio (active/total) increased from day 16 to days 18-22 and then decreased to the same level as found during the suckling period. At the onset of weaning the active form of phosphorylase decreased, whereas total phosphorylase initially increased and then decreased after postnatal day 20. Both forms of phosphorylase increased again at the end of the weaning period. The activity ratio decreased at the start of weaning and remained low throughout the rest of the weaning period. The effects of premature weaning were similar to those observed in normally weaned animals, but the changes occurred sooner and were more pronounced. PMID- 2985220 TI - Benzodiazepine receptors and seizure susceptibility in epileptic fowl. AB - Benzodiazepine binding to brain membrane preparations obtained from epileptic and nonepileptic carrier fowl was compared. [3H]Flunitrazepam binding to whole brain homogenates from 2-day-old chicks and [3H]diazepam binding to synaptosomal membranes and homogenates from adult chickens were determined. Scatchard analysis revealed no differences in either the number of receptors or their affinity for the ligands when the epileptics were sacrificed in the interictal state. Evoked seizures in adult epileptics had no effect on the number or affinity of binding sites using [3H]diazepam as the ligand. Moreover, the ability of gamma aminobutyric acid to facilitate benzodiazepine binding was not different in epileptic fowl when compared with carriers. PMID- 2985221 TI - Oxygen radicals in the adult respiratory distress syndrome, in myocardial ischemia and reperfusion injury, and in cerebral vascular damage. AB - Recent work suggests that oxygen radicals may be important mediators of damage in a wide variety of pathologic conditions. In this review we consider the evidence supporting the participation of oxygen radicals in the adult respiratory distress syndrome, in ischemia reperfusion injury in the myocardium, and in cerebral vascular injury in acute hypertension and traumatic brain injury. In the adult respiratory distress syndrome there is active sequestration of polymorphonuclear neutrophils in the pulmonary vascular system. There is evidence that activation of these neutrophils results in the production of oxygen radicals which injure the capillary membrane and increase permeability, leading to progressive hypoxia and decreased lung compliance which are hallmarks of the syndrome. In acute arterial hypertension or experimental brain injury oxygen radicals are important mediators of vascular damage. The metabolism of arachidonic acid is the source of oxygen free radical production in these conditions. In myocardial ischemia and reperfusion injury, the ischemic myocyte is "primed" for free radical production. With reperfusion and reintroduction of molecular oxygen there is a burst of oxygen radical production resulting in extensive tissue destruction. Myocardial ischemia--reperfusion injury shares in common with the other two syndromes activation of the arachidonic acid cascade and acute inflammation. Thus it would appear that the generation of toxic oxygen species may represent a final common pathway of tissue destruction in several pathophysiologic states. PMID- 2985222 TI - Renal responses to atrial natriuretic factor during converting enzyme inhibition. AB - The effect of converting enzyme inhibitor (CEI) on the renal response to atrial natriuretic factor (ANF) was determined in the rat. In the absence of CEI, ANF produced rapid and significant increases in sodium, potassium, calcium, and urine excretions while blood pressure declined transiently. In the presence of CEI, ANF enhanced the excretion of sodium and potassium but not of calcium and urine. The activity of CEI was documented by observing that, in the presence of CEI, the elevation of blood pressure produced by angiotensin I was significantly attenuated. The potentiating effect of CEI on the natriuretic response to ANF supports the hypothesis that converting enzyme may be involved in the metabolism of ANF. PMID- 2985224 TI - Prolactin as a luteotrophin. AB - This review summarizes evidence suggesting a direct luteotrophic role for the hypophyseal hormone prolactin (PRL). This direct role consists of the capability to stimulate progesterone synthesis in vitro, the capability to maintain the membrane fluidity and receptors for luteinizing hormone and the capability to import substrate for progesterone synthesis. The time required for PRL-induced luteotrophic events is in the order of hours and sometimes days, and it appears that the effects are not associated with acute intracellular changes. The relatively slow responses and the stimulation of specific protein synthesis by PRL in target tissues other than the ovary suggest that PRL may function primarily through activation of the genome. PRL may induce the synthesis of specific luteal proteins, including enzymes for the regulation of intracellular substrate pools, membrane receptors for LH, or receptor proteins for lipoproteins, a major extracellular source of substrate. PMID- 2985223 TI - The regulation of steroidogenesis is different in the two types of ovine luteal cells. AB - Ovine luteal tissue contains two distinct steroidogenic cell types, small (8-20 microns) and large (greater than 20 microns), which differ based on morphological and biochemical criteria. Unstimulated small cells secrete low levels of progesterone, respond to LH or dibutyryl cAMP (dbcAMP) with enhanced secretion of progesterone, and contain most of the receptors for LH. The unstimulated large cells, conversely, secrete high levels of progesterone, have few, if any, receptors for LH, and do not respond to LH or dbcAMP with increased progesterone secretion. The lack of response to dbcAMP by large cells was investigated. Large cells incubated in the presence of cholesterol, ram serum, or 25 hydroxycholesterol did not demonstrate substrate limitation. Hormone-independent stimulation of adenylate cyclase by cholera toxin or forskolin resulted in increased adenylate cyclase activities (P less than 0.01), cAMP accumulation (P less than 0.05), and the binding of endogenous cAMP (P less than 0.05) by type I cAMP-dependent protein kinase in both small and large cells. These treatments were accompanied by enhanced secretion of progesterone (P less than 0.05) in small cells. In contrast, large cells did not respond with an increase in progesterone secretion under these conditions. These observations suggest that the high rate of secretion of progesterone in unstimulated large cells is not regulated by cAMP. PMID- 2985225 TI - The role of plasma lipoproteins in steroidogenic response of rat luteal cells during gonadotropin-induced refractory states. AB - Administration of human chorionic gonadotropin (hCG) to pregnant mare's serum gonadotropin--hCG primed rats results in the loss of in vitro responsiveness of the ovaries to exogenous gonadotropins for progesterone production. This state is associated with a loss of membrane receptors for hCG and a concomitant increase in lipoprotein receptors. Although lipoproteins potentiated gonadotropin response in ovaries from saline-injected rats, no stimulation was observed in hCG desensitized ovarian cells. Examination of the time course for the loss of lipoprotein response after hCG injection revealed that injection with 50 IU of hCG results in a loss of gonadotropin response as early as 1 h after injection, but exogenous cholesterol-carrying lipoprotein fractions, LDL and HDL, were capable of stimulating progesterone production up to 4 h after hormone injection. Measurement of endogenous cholesteryl ester content showed that there was a 72% decline during this period with a concomitant increase in the basal progesterone production. One hour after hCG injection there was no stimulation of steroidogenesis by hCG in the presence or absence of exogenous lipoproteins. The refractoriness to exogenous hCG appeared only 4 h later when the hCG dose was reduced to 10 IU, whereas with 25 IU of hCG, the effect was similar to that observed using 50 IU of hCG. Such diverse steroidogenic stimuli as hCG, LH, LDL, cAMP, and cholera enterotoxin failed to stimulate progesterone synthesis in vitro in luteal cells of rats injected with 50 IU of hCG 48 h prior to sacrifice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985226 TI - Seasonal distribution of enteroviruses and adenoviruses in domestic sewage. AB - A seasonal distribution of enteroviruses and adenoviruses in raw sewage effluents of Athens, Greece, was observed over a 15-month surveillance period. All 36 samples tested were positive for both virus groups. Adenovirus concentration levels ranged from 70 to 3200 cytopathic units per litre of sample, whereas the corresponding values for enteroviruses were 90-900 cytopathic units per litre. Peak values for adenoviruses were recorded during the months of April and June 1983, whereas for enteroviruses the peak was recorded in September 1983. All three types of poliovirus were present. Coxsackievirus types B-1, B-2, B-4, and B 5 and echovirus type 7 were also isolated. Adenovirus types 1, 2, 3, 5, 7, and 15 were detected as well. PMID- 2985227 TI - Long-term survival of human rotavirus in raw and treated river water. AB - This study was aimed at assessing the role of water as a vehicle for rotavirus spread by determining how well these viruses survive in the water environment. A cell culture adapted strain of human rotavirus subgroup 2, grown in MA-104 cells, was used as a model. Virus survival was tested in the following types of water samples, derived from the Ottawa River, at two different times of the year: (i) raw water (RW), (ii) muncipally treated tap water (TW), and (iii) raw water that had been filtered (FW) through a membrane (0.22 micron). The water samples, with approximately 5.0 X 10(4) plaque-forming units (PFU) of the virus, were held at either 4 or 20 degrees C and tested for infectious virus over a period of 64 days. The TW samples had a total and free chlorine content of 0.05 and less than 0.05 mg/L, respectively. The chlorine in these samples was not neutralized before virus contamination. Irrespective of the holding temperature, the virus titre in FW remained essentially unaltered throughout the test period. In TW held at 4 degrees C, there was no significant drop in the virus titre even after 64 days, whereas at 20 degrees C the titre in TW was reduced by about 2 log10 over the same period. Even though the loss of virus infectivity was most rapid in RW held at 20 degrees C, it took about 10 days for a 99.0% reduction in the plaque titre of the virus. These findings, therefore, indicate that rotaviruses can survive for several days in raw and treated river water thus making recreational and potable waters potential vehicles for the transmission of rotavirus infections. PMID- 2985228 TI - Detection of collagenase activity in oral bacteria. AB - Collagenolytic activity of 12 species of oral bacteria was assessed using two methods of detection. Except for two species, all bacterial strains tested were capable of degrading at least one general protein substrate. Results of collagenolytic activity in a growth assay indicate that Bacteroides gingivalis is the only bacterium capable of degrading collagen when the substrate is sterilized using ethylene oxide. However, if the substrate is sterilized by autoclaving, in the presence or absence of the growth medium, other bacterial species could be shown to be collagenolytic. Collagenolytic activity was also demonstrated when whole or broken cells were used in a [14C]collagen assay. Results from this assay and from inhibition studies indicate that collagenolytic activity can either be the result of the combined activities of both a specific collagenase and nonspecific proteases (B. gingivalis) or nonspecific proteases only (other strains in this study), although in the latter case, the time taken to hydrolyze collagen can be 10 times longer than with a specific collagenase. PMID- 2985231 TI - Viruses, genes, and cancer. I. The discovery of cellular oncogenes and their role in neoplasia. PMID- 2985230 TI - Human T-cell leukemia (lymphotropic) retroviruses and their causative role in T cell malignancies and acquired immune deficiency syndrome. PMID- 2985229 TI - Polyclonal activators in pulmonary immune disease. PMID- 2985232 TI - Ovarian dysfunction in patients with gestational trophoblastic neoplasia treated with short intensive courses of etoposide (VP-16-213). AB - The effect of oral Etoposide (VP-16-213) on the ovarian function in 22 patients with residual gestational trophoblastic disease was studied by serial weekly measurement of serum follicle stimulating hormone (FSH), 17 beta-estradiol (E2), progesterone (P) and prolactin (PRL), and monitoring of the menstrual patterns during and after drug treatment. Multiple courses of VP-16-213 at a dosage of 200 mg/m2 X 5 days were given 1 week apart. Following an average dose of 5.0 gm, five young patients developed transient ovarian failure for a mean period of 2.4 months and two elderly patients developed permanent ovarian failure (total incidence, 7/22, 31.8%) as evidenced by hypergonadotropic, hypoestrogenic amenorrhea. In addition, two patients each developed anovulatory cycles and hypomenorrhea. VP-16-213 should therefore be regarded a potent gonadotoxic drug and these hitherto unreported side effects should be recognized when the drug is used in menstruating females. PMID- 2985233 TI - Is mallory body formation a preneoplastic change? A study of 181 cases of liver bearing hepatocellular carcinoma and 82 cases of cirrhosis. AB - The hypothesis that Mallory body formation by hepatocytes is a sign of preneoplasia was tested. This hypothesis was based on animal experiments but has not been tested in man. The authors studied the livers of 181 human autopsies in which hepatocellular carcinoma (HCC) was present and 82 cirrhotic livers from patients with alcoholism, HB viral infection, or cryptogenic cirrhosis. The frequency of Mallory bodies in nonneoplastic hepatocytes was 40% in the HCC bearing livers with cirrhosis (LC). In HCC-bearing livers with pre-cirrhotic changes (PC), 25% showed Mallory body formation by nonneoplastic hepatocytes. In the cases of HCC, where there was no accompanying PC or LC, Mallory bodies were never found in the nonneoplastic hepatocytes. When the 82 cirrhotic livers without HCC and the 116 cirrhotic cases with HCC were combined, it was found that HCC was present in 70% of cirrhotic livers when the nonneoplastic liver cells contained Mallory bodies. When no Mallory bodies were found in the nonneoplastic liver cells, HCC was present in 53% of cases. The difference between the two groups was significant (P less than 0.05). The difference was significant for both HB viral hepatitis and cryptogenic cirrhosis but not for alcoholic cirrhosis. Likewise, when nonneoplastic hepatocytes formed Mallory bodies in cirrhotic livers, there was a statistically significant increase in the number of HCC cells that formed Mallory bodies (P less than 0.01). When nonneoplastic hepatocytes occurred in groups of Mallory body forming cells, the hepatocellular features were atypical and characteristic of dysplastic cells. The evidence indicates that when Mallory body formation was observed in HBsAg-positive and cryptogenic cirrhotic livers, they were associated with an increased frequency of HCC formation in man. PMID- 2985234 TI - Cellular vitamin A-binding proteins in liver tumors and adjacent tissues. AB - The levels of cellular retinol-binding protein (CRBP) and retinoic acid-binding protein (CRABP) were assessed in surgically resected ten human liver tumors and their adjacent tissues by sucrose density gradient centrifugation and Scatchard analysis. The tissues adjacent to hepatocellular carcinomas usually showed portal cirrhosis or hepatitis. There was no significant difference in the dissociation constant (Kd) values of binding proteins of CRBP or CRABP between tumors and adjacent tissues. Five of the ten liver tumors showed similar levels of CRBP as in their adjacent tissues whereas three of the ten liver tumors showed about 40% or less contents of CRBP in comparison with their adjacent tissues and two others had no CRBP activity. By contrast, CRABP activities were found in five liver tumors and in adjacent tissues of one case, whereas there were no detectable CRABP activities in the adjacent tissues except in one case. PMID- 2985235 TI - Induction of unusual pancreatic neoplasms, with morphologic similarity to human tumors, and evidence for their ductal/ductular cell origin. AB - A variety of pancreatic neoplasms, including giant cell, poorly differentiated or other unusual types, some morphologically similar to those observed in humans were induced in Syrian hamsters. Unclassified or anaplastic forms were also noted in the hamster. The finding of a blood group A-like antigen, which is produced by normal pancreatic ductal and ductular cells, in all of these induced tumors, but none in acinar, insular, and mesenchymal cells, suggests their ductal/ductular cell origin. PMID- 2985236 TI - Involvement of nipple and areola in early breast cancer. AB - In 141 mastectomy specimens, performed for invasive or noninvasive carcinomas, histopathologic study was performed to assess the extent of nipple-areola involvement by the tumor. In this study, patients were excluded if the tumor was located beneath the areola; and nipple and/or areola abnormalities were clinically present. Tumor involvement of the nipple and/or areola was found in 44 of 141 specimens (31%), with intraductal growth in 36 (82%) of 44, stromal invasion in 3 (7%), and ductal and stromal invasion in 5 (11%). Analysis of nipple-areolar involvement with consideration of the different variables indicates that it occurred in association with tumor size, tumor-areola distance, and histologic type. Such information provides clinically relevant guidelines in decision making for limited breast surgery. PMID- 2985237 TI - Cytogenetic studies of stimulated lymphocytes in hairy cell leukemia. AB - Using a sister chromatid differentiation (SCD) technique, cell cycle analysis in lymphocytes from two patients with hairy cell leukemia (HCL) revealed it to be similar to cell cycle progression of normal lymphocytes stimulated with lipopolysaccharide W from Escherichia coli 0.55:B5 (LPS). It appears that LPS can readily stimulate the leukemic cells of HCL into mitosis. In the two cases of B cell HCL studied, one (case 1) was revealed to have an abnormal clone with a missing chromosome #22 that was related to the production of lambda-chains. PMID- 2985238 TI - Defective interleukin-1 production by natural killer cells of patients with cancer. AB - Large granular lymphocytes (LGLs) from patients with malignant disease and from controls were activated by endotoxin or K562 cells, and the supernatants assayed for interleukin-1 (IL-1) activity. Normal LGLs produced significant amounts of IL 1, the activity of which could be neutralized by anti-human IL-1 antiserum. In patients with advanced cancer depressed IL-1 production was observed, which generally correlated with the degree of cytotoxicity produced by the LGLs. Prior treatment of the LGLs with interferon increased production of IL-1 by both control and patient cells. It is suggested that LGLs coming into contact with K562 cells produce IL-1, which is important in the effector-target cell interaction. The decreased cytotoxic activity of LGLs from cancer patients could be related to a defect in IL-1 production, an effect which can be partially corrected by in vitro interferon treatment. PMID- 2985239 TI - Elimination of small cell lung cancer cells in vitro from human bone marrow by a monoclonal antibody. AB - We report here a useful method for elimination of small cell lung cancer cells in vitro from bone marrow. A monoclonal antibody, TFS-2, which mediates complement lysis and recognizes an antigen present on small cell lung cancer cells but not lymphoid cells or bone marrow cells, was used to clear infiltrated bone marrow. The antibody in the presence of complement effectively killed tumor cells, but it was not cytotoxic to bone marrow cells. When mixed populations consisting of tumor cells and bone marrow cells were treated with antibody and complement, the tumor cells were also effectively killed, except when large numbers of bone marrow cells were present, whereas TFS-2 had no significant effect on bone marrow stem cells, as judged by colony-forming unit assays. PMID- 2985240 TI - Requirements for and kinetics of growth arrest of neoplastic cells by confluent 10T1/2 fibroblasts induced by a specific inhibitor of cyclic adenosine 3':5' phosphodiesterase. AB - This study was conducted to further characterize the previously described phenomenon of growth inhibition of neoplastically transformed C3H/10T1/2 cells (T10T1/2) by nontransformed C3H/10T1/2 clone 8 mouse embryo fibroblast (10T1/2) cells in the presence of inhibitors of cyclic adenosine 3':5'-monophosphate (cAMP) phosphodiesterase. The cAMP phosphodiesterase inhibitor dl-4-(3-butoxy-4 methoxybenzyl)-2-imidazolidinone (RO20-1724) was shown to be completely nontoxic to T10T1/2 cells at 10(-4) M, yet when added to mixed cultures of T10T1/2 cells and postconfluence growth-arrested 10T1/2 cells, colony formation and [3H]thymidine incorporation into T10T1/2 cells were virtually eliminated. This effect was dose dependent and was reversible upon drug withdrawal. In 10T1/2 cells, RO20-1724 caused a dose-dependent increase in cAMP levels from about 5 to 150 pmol/10(6) cells; in T10T1/2 cells, 10(-4) M drug treatment caused a 5-fold elevation in cAMP without a clear dose dependency. Cyclic guanosine 3':5' monophosphate levels in 10T1/2 cells fell by 50% with drug treatment but were unmeasurable in T10T1/2 cells. When intracellular cyclic AMP levels were elevated by the adenyl cyclase stimulator forskolin, growth inhibition of T10T1/2 cells was again induced in mixed cultures but was not observed when added to T10T1/2 cells alone. Addition of RO20-1724 to low concentrations of forskolin produced a greater than additive effect on growth inhibition. Growth inhibition of T10T1/2 cells by RO20-1724 was shown to (a) require contact with, or extremely close proximity to, a confluent monolayer of 10T1/2 cells, (b) be maximum when seeded upon a growth-inhibited monolayer and not an actively growing 10T1/2 culture, and (c) not be decreased by continuous agitation of the culture medium, indicating that readily diffusible inhibitory factors are not involved. A model is presented whereby transformed cells can respond to but cannot themselves generate growth inhibitory signals produced by post-confluence growth-inhibited nontransformed cells. The existence of these cellular interactions may well explain problems in the quantitation of transformed foci encountered in the use of this cell line as an assay system for chemical and physical carcinogens. PMID- 2985241 TI - Cull rates of dairy cattle with antibodies to bovine leukemia virus. AB - The relationship of cull rate to age was investigated retrospectively in dairy cows with and without antibodies to bovine leukemia virus (BLV). Banked sera from eight annual herd tests on one 200-cow herd were tested for presence of BLV antibodies by agar-gel immunodiffusion using the Mr 51,000 glycoprotein antigen of BLV. Age-specific cull rates were computed for BLV-antibody-positive and antibody-negative cows yearly from 2 to 7 years of age. Cull rates, transformed by the Arc-sin square root, were analyzed by weighted regression. Transformed cull rates increased significantly as BLV-antibody-positive cows aged (one-tailed P = 0.023) but not as antibody-negative cows aged (one-tailed P = 0.59). A Mantel Byar survival analysis showed significantly longer survival beyond 3.5 years of age among antibody-negative cows than among antibody-positive cows (P = 0.008). PMID- 2985242 TI - Lysis of tumor cells by human blood monocytes by a mechanism independent of activation of the oxidative burst. AB - Tumorilytic human blood monocytes recognize and destroy neoplastic cells by a mechanism that is nonphagocytic and requires cell-to-cell contact. The mechanism of cytolysis subsequent to binding is controversial. Release of reactive oxygen intermediates by activated rodent macrophages has been suggested as an important mechanism for tumor cell lysis in some short-term cytotoxicity assays. We examined whether oxygen intermediates are also responsible for mediating the lysis of adherent human tumor cells in a long-term (72-h) tumoricidal assay. Human blood monocytes were incubated with medium, concanavalin A-stimulated lymphokine [macrophage-activating factor (MAF)], lipopolysaccharide endotoxin, or human recombinant gamma interferon for 24 h prior to the addition of [125I] iododeoxyuridine-labeled A375 melanoma cells. The following evidence indicated that monocyte-mediated tumor cell lysis was independent of superoxide anion (O2-) and H2O2 production: (a) although human blood monocytes incubated for 24 h with gamma interferon produced twice as much O2- as control or MAF-treated monocytes, gamma interferon did not activate monocyte tumoricidal activity unless combined with lipopolysaccharide endotoxin, 0.2 ng/ml or more; (b) incubating the monocytes with 10 nM phorbol myristate acetate for 0.5 h stimulated O2- production but no cytotoxicity; (c) the cytolytic activity of MAF-treated monocytes was not decreased in the presence of catalase or superoxide dismutase; and (d) finally, peripheral blood monocytes were isolated from six patients with chronic granulomatous disease, activated by MAF or lipopolysaccharide endotoxin, and then assayed for tumoricidal activity. While these activated chronic granulomatous disease monocytes did not produce O2- or H2O2, tumor cell lysis was normal in all six patients. Hence, lysis of tumor cells in a 72-h assay is not dependent upon the generation of O2- and/or H2O2 and is intact in chronic granulomatous disease monocytes. PMID- 2985243 TI - Multistep change in epidermal growth factor receptors during spontaneous neoplastic progression in Chinese hamster embryo fibroblasts. AB - Whole Chinese hamster embryo lineages have been shown to undergo multistep spontaneous neoplastic progression during serial passage in culture. We have studied the binding, internalization, and degradation of 125I-labeled epidermal growth factor at four different stages of transformation. The whole Chinese hamster embryo cells lost cell surface epidermal growth factor receptors gradually during the course of neoplastic progression until only 10% of the receptor number present in the early-passage cells (precrisis) were retained in the late-passage cells (tumorigenic). No differences in internalization rates, chloroquine sensitivity, or ability to degrade hormone between the various passage levels were seen. No evidence for the presence in conditioned medium of transforming growth factors which might mask or down-regulate epidermal growth factor receptor was obtained. These results suggest that a reduction in cell surface epidermal growth factor receptor might be an early event during spontaneous transformation in whole Chinese hamster embryo cells. PMID- 2985245 TI - Inhibitors of poly(adenosine diphosphoribose) synthetase, examination of metabolic perturbations, and enhancement of radiation response in Chinese hamster cells. AB - 3-Aminobenzamide, a specific inhibitor of poly(adenosine diphosphoribose) synthesis, has been shown to enhance the response of mammalian cells to ionizing radiation and alkylating agents. Observations such as these usually have been taken to be an indication of the involvement of poly(adenosine diphosphoribose) in the repair of DNA damage. It has been reported that some inhibitors of adenosine diphosphoribosyl transferase (ADPRT) affect cell viability, glucose metabolism, and DNA synthesis when present at low concentrations in the growth medium for extended periods (e.g., lymphoid cells exposed to a few millimolar for 24 h [Milam, K. M., and Cleaver, J. E. Science (Wash. DC), 223: 589, 1984]). The latter report questioned previous interpretations of radiation results based on the use of ADPRT inhibitors which enhance cell killing. We have studied the enhanced radiation lethality of Chinese hamster cells using higher concentrations of these inhibitors, but for shorter periods, in an effort to determine if metabolic perturbations are produced and if they are relatable to enhanced cell killing. The compounds used were 2-aminobenzamide, 3-aminobenzamide, 4 aminobenzamide, benzamide, and nicotinamide, compounds which show a large variation in their potency as inhibitors of ADPRT. It was found that none of the compounds was toxic at the highest doses used (20 mM for 2 h) and that, during a 2-h period, the potent inhibitor 3-aminobenzamide had little or no effect on DNA synthesis. Two h is long enough to yield a near-maximum radiosensitization with 3 aminobenzamide. Although glucose metabolism was affected to varying degrees (up to a 50% inhibition by 4-aminobenzamide in 2 h), there was no correlation between effectiveness in inhibiting ADPRT and effectiveness in inhibiting glucose metabolism. A correlation was observed, however, between the inhibitory potential of ADPRT and the enhancement of radiation response. When used for sufficiently short times, we conclude that the effects at even high concentrations of a potent inhibitor of ADPRT (e.g., 3-aminobenzamide) are consistent with an involvement of poly(adenosine diphosphoribose) synthesis in the expression of a radiation induced end point like cell killing. PMID- 2985244 TI - Control of Polyomavirus T-antigen and DNA synthesis in mouse embryo fibroblast cells by vitamin A. AB - Vitamin A (retinoic acid, 10(-6) M) treatment of confluent mouse embryo cells for only 7 h resulted in optimal inhibition of Polyomavirus replication. Depending upon the input multiplicity of virus, one could wait until between 12 and 18 h postinfection to add vitamin A and still observe maximal inhibition of virus yields. Taken together, and assuming the same kinetics before and after virus infection, these results suggested that the inhibitory action of vitamin A occurred between 19 and 25 h into the Polyomavirus replication cycle. In this model system, such a time corresponded to the onset of T-antigen expression and virus-induced cellular DNA synthesis. Analysis of both viral and virus-induced cellular DNA synthesis by the method of Hirt (J. Mol. Biol., 26: 365-369, 1967) and by cesium chloride gradients suggested that vitamin A preferentially inhibited viral, more than virus-induced cellular, DNA synthesis in confluent cell monolayers. Vitamin A also concomitantly inhibited Polyomavirus T-antigen expression in such confluent cultures. In contrast, viral DNA synthesis and infectious virus yields were not significantly inhibited by vitamin A in subconfluent cell cultures. The antagonistic effect of vitamin A on Polyomavirus replication in confluent monolayers could be blocked with cycloheximide, a reversible protein synthesis inhibitor. This suggested that vitamin A inhibition of Polyomavirus replication was indirect and mediated by a newly synthesized protein. Taken together, these results suggest that vitamin A induced a protein in confluent, but not subconfluent, cells, which blocked the expression of Polyomavirus T-antigen. Decreased amounts of T-antigen most likely reduced Polyomavirus and cellular DNA synthesis and virus yield. PMID- 2985246 TI - Formation of cadaverine as an effect of alpha-difluoromethylornithine on chick embryo fibroblasts transformed by rous sarcoma virus. AB - Chick embryo fibroblasts grew normally in the presence of 1 X 10(-3) to 10 X 10( 3) M alpha-difluoromethylornithine (DFMO). This drug did not interfere with protein and DNA synthesis of normal fibroblasts and of cells transformed by Rous sarcoma virus. The morphological appearance of normal and transformed cells was not altered by DFMO, as determined by scanning electron microscopy. Flow microfluorometric analyses also confirmed the notion that normal or transformed cells were not blocked by DFMO in the G1 phase of the cell cycle. As expected, DFMO reduced cellular putrescine levels. This diamine, however, was replaced by the analogue cadaverine (1,5-diaminopentane), which accumulated mainly in the transformed cells. The increase in cellular cadaverine levels was also demonstrated during the infection of chick embryo fibroblasts with a temperature sensitive mutant of Rous sarcoma virus under permissive conditions. Under restrictive conditions (42 degrees C), less cadaverine accumulated in the infected cells. These findings suggest that diamines and polyamines are necessary for the transformation process and that blocking one pathway by DFMO leads to the activation of an alternative biosynthetic pathway. PMID- 2985248 TI - Coordinate inhibition of plasminogen activator and tumor growth by hydrocortisone in mouse mammary carcinoma. AB - A body of evidence has suggested that hormones which modulate plasminogen activator production by cultured tumor explants in vitro may have a qualitatively comparable effect on the growth of the same tumors in vivo. As a test of this correlation and to explore its potential for predicting the in vivo response of tumors to hormones, we have studied here the effect of hydrocortisone on the growth of primary and first generation transplants of mouse mammary tumor virus determined mammary tumors in BALB/c X DBA/8 F1 (hereafter called CD8F1) mice; hydrocortisone had been found previously to inhibit plasminogen activator production by explants of these tumors. The results were: (a) hydrocortisone reversibly blocked the growth of palpable primary tumors; growth resumed at control rates following withdrawal of exogenous hormone; (b) hydrocortisone inhibited the growth of first-generation tumor transplants when administered either before or after the appearance of palpable tumors; (c) pretreatment with hydrocortisone both delayed the appearance of primary tumors and greatly reduced tumor incidence in susceptible mice; a substantial part of the decrease in tumor incidence was apparently irreversible; (d) hydrocortisone reduction of tumor growth was accompanied by inhibition of tumor plasminogen activator content, and these effects displayed a similar dose dependence (enzyme content of tumor lysates was measured by the 125I-fibrin plate assay); the enzyme present in control and hormone-treated tumors was predominantly of the urokinase type. These findings suggest that plasminogen activator production and mammary tumor growth in CD8F1 mice are coordinately regulated and thus encourage the view that plasminogen activator might be useful as an in vitro marker for predicting the in vivo response of tumors to hormones. PMID- 2985247 TI - Carcinogenesis by nonmutagenic chemicals: early response of rat liver cells induced by methapyrilene. AB - Although methapyrilene (MP) produces hepatocellular carcinomas in rats, it does not elicit many of the cellular responses induced by other hepatocarcinogens. We have investigated the early changes induced in rat liver epithelial cell cultures by MP using morphological, cytochemical, and cytofluorometric techniques. Within 2 h of MP treatment, inclusion bodies which were stainable with lipid stains were observed in the cytoplasm. Ultrastructurally, they resembled lamellar bodies with alternating light and dark lamellae. These bodies were transient in nature, since they disappeared within 24 h of removal of MP. They were, however, retained in the cytoplasm as long as MP was present in the medium. Lamellar bodies appear to be induced in the presence of histamine H1 receptor-blocking agents, since methaphenilene and diphenhydramine produced this reaction, but cimetidine, an H2 antagonist, did not. Morphologically, mitochondria of control cells were long and rod-like, whereas they were short or bizarrely shaped in the MP-treated cells. Moreover, a quantitative increase was observed in the mitochondrial content of the treated cells, when intact liver cells vitally stained with Rhodamine 123 were analyzed with a fluorescence-activated cell sorter. A significant increase in binucleated cells was observed when liver cells were exposed for 10 to 12 days with MP. Collectively, these results suggest that MP might perturb the cytoskeletal elements leading to an alteration in the nuclear and mitochondrial makeup of rat liver cells. PMID- 2985249 TI - Relationship of histology of Wilms' tumor to growth characteristics of nude mouse heterotransplants. AB - Eighteen Wilms' tumors (WIT), including classical triphasic WIT (blastema, tubules, and mesenchyme) and WIT variants (blastema and tubules, monomorphous tubules, multiloculated cysts, rhabdomyomatous WIT, and clear cell sarcoma), were heterotransplanted in nude mice. Ten (56%) tumors grew and were serially passaged. With two exceptions, the histology of the surgically resected tumors and heterotransplants was found to be similar. Tumors showing a prominent blastema component grew rapidly, whereas those with tubular epithelial or mesenchymal differentiation grew more slowly. Tumors injected s.c. consisted almost entirely of blastema, while tumors injected i.p. consisted of blastema with large areas of tubular epithelium. These results demonstrate that nude mouse heterotransplants of WIT closely resemble the surgically resected tumors from which they derive, that growth rates of WIT heterotransplants depend on the identity of the tumor cells, and that differentiation of WIT heterotransplants can be modulated, depending on the route of administration of tumor cells. PMID- 2985250 TI - Induced secretion of a Mr 46,000 protein by cultured human hepatoma cells treated with tumor promoters. AB - Human hepatoma cells, HuH-6 Cl-5, were treated with 12-O-tetradecanoylphorbol-13 acetate at concentrations of 1 ng/ml to 10 micrograms/ml for 6 h in the presence of [35S]methionine. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the labeled proteins secreted into the medium showed that this treatment induced marked secretion of a polypeptide with a molecular weight of about 46,000 (p46). When the labeled proteins were analyzed by two-dimensional polyacrylamide gel electrophoresis, p46 was composed of three isoproteins which had different isoelectric points. The two new classes of tumor promoters, teleocidin and aplysiatoxin, also induced secretion of this protein, although the amount of p46 induced by debromoaplysiatoxin was less than that induced by 12-O tetradecanoylphorbol-13-acetate, teleocidin, and aplysiatoxin, judging from densitometric scanning of bands on a fluorogram. The nonpromoting phorbol esters, such as 4 beta-phorbol and phorbol-13-monoacetate, did not induce p46. The induction of p46 secretion involved de novo RNA synthesis, since actinomycin D (1 microgram/ml) completely and selectively blocked the incorporation of [35S] methionine into the protein. "Pulse-chase" experiments indicated that p46 was not a degradation product induced by these potent tumor promoters. In an attempt to identify p46, the total proteins released were treated with two kinds of rabbit anti-human whole serum antisera, but although some proteins were precipitated, p46 was not. PMID- 2985251 TI - Stability and utility of the unique human small cell carcinoma line SHP-77. AB - The human small cell (oat cell) carcinoma line, SHP-77, established by Fisher and Paulson in 1977 and originally described as a "large cell variant of oat cell cancer" has been evaluated by several different parameters and shown even after more than 200 passages to retain properties described for the original cell line. Karyotypic, histological, and biochemical features are retained, as well as tumorigenicity in nude mice. The original authors' suggestion that this is a propitious cell line for both in vitro and in vivo studies is supported by this report. Modulation of growth characteristics in vivo (in xenografts) emphasizes the plasticity of this unique line which serves as a valuable model for basic as well as therapeutic studies. SHP-77 can serve as an in vitro target in 51Cr and 111In release cytotoxicity assays as well as in in vivo nude mouse assays for evaluating immune reactivity of cells and serum from lung cancer patients. The potential histological variability of SHP-77, despite its biochemical stability, calls attention to the inadequacy of histological criteria for lung tumor classification. PMID- 2985252 TI - Inhibition of tumor cell growth by aggregation of a tumor-associated glycolipid antigen: a close functional association between gangliotriaosylceramide and transferrin receptor in mouse lymphoma L-5178Y. AB - The growth of mouse lymphoma L-5178Y cells with a high degree of gangliotriaosylceramide expression (high-expressor clone AA12) was inhibited by the addition of a biotinylated mouse immunoglobulin M monoclonal antibody (2D4) directed to gangliotriaosylceramide followed by cross-linking with avidin or a second antibody (anti-mouse immunoglobulin M). This growth inhibition was observed in both serum-containing medium and a chemically defined medium containing transferrin as the only growth factor. Cell growth was not inhibited by the addition of biotinylated antibody 2D4 alone, avidin alone, or the biotinylated derivative of an immunoglobulin M monoclonal anti-N acetyllactosamine antibody (1B2) cross-linked with avidin. The growth of lymphoma L-5178Y 27AV cells, which do not express gangliotriaosylceramide (non-expressor clone), was not inhibited by either of these monoclonal antibodies or their biotinylated derivatives plus avidin. In the presence of biotinylated antibody 2D4 and avidin, cells of the high-expressor clone (L-5178Y AA12) displayed a capping of gangliotriaosyl antigen. In contrast, the transferrin receptor and the major glycoproteins (concanavalin A receptors) were not capped in the presence of biotinylated antibody 2D4 and avidin but were homogeneously distributed on the cell surface. Cells whose growth was inhibited by the addition of biotinylated antibody 2D4 and avidin showed an inhibition of 125I-transferrin internalization, although binding of 125I-transferrin to the cell surface was similar to that of control cells. These results indicate that the tumor antigen gangliotriaosylceramide is functionally associated with the transferrin receptor and may regulate the process of internalization of transferrin. PMID- 2985253 TI - Proviral structure and differentiation antigen phenotype of spontaneous and chemically induced AKR lymphomas. AB - AKR mice develop spontaneous thymomas after 6 months of age due to a novel class of murine leukemia viruses that are generated by a series of genetic recombinations between endogenous proviral loci. AKR mice also are more susceptible to N-methyl-N-nitrosourea (MNU)-induced thymomas than are low leukemia-incidence mouse strains. To determine whether virally and chemically induced lymphomagenesis proceeds by similar pathways in AKR mice, spontaneous and MNU-induced thymic lymphomas were analyzed for a DNA restriction linkage generated during spontaneous tumor development by recombination between envelope genes of endogenous murine leukemia proviral loci. DNA from spontaneous thymic lymphomas invariably contained a restriction fragment characteristic of recombinant murine leukemia virus etiology, while four of five MNU-induced thymic lymphomas did not show this restriction linkage. In addition, analysis of lymphocyte differentiation antigen profiles indicated that MNU-induced lymphomas represent a more immature stage of T-cell differentiation than the majority of spontaneous lymphomas. These data suggest that there are fundamental differences in the mechanisms of induction of virally and chemically induced thymic lymphomas in AKR mice. PMID- 2985255 TI - Human T-cell leukemia-lymphoma virus type 1 and adult T-cell leukemia-lymphoma in Okinawa. AB - A serological survey for the presence of antibodies against the human T-cell leukemia virus, type 1 (HTLV-1) in patients seen at the Chubu Hospital in Okinawa was undertaken. All patients with the clinicopathological diagnosis of adult T cell leukemia-lymphoma were positive. These cases had the characteristic features of adult T-cell leukemia-lymphoma: diffuse histology, often mixed cell or pleomorphic, and a high frequency of hypercalcemia, leukemic phase, diffuse visceral involvement, and opportunistic infections. The median survival of these patients was short, being only 18 weeks. Of the other patients with cancers screened, two of five other non-Hodgkin's lymphoma patients were positive and three of eight patients with other hematological cancers were positive. In addition, three of the four immediate family members of one adult T-cell leukemia lymphoma case had antibodies. Of the other persons (both in- and outpatients) without hematological cancers, those under the age of 50 had a much lower antibody prevalence (4%) than those over 50 (30%). There was no significant difference in antibody prevalence between the two sexes in either the younger or older age group. These findings further document that Okinawa is an endemic area for HTLV-1. None of the 157 individuals screened for antibodies to HTLV-3 were positive, consistent with the fact that no cases of the acquired immune deficiency syndrome have been reported from Okinawa. PMID- 2985254 TI - Growth of human lung tumor in the brain of the nude rat as a model to evaluate antitumor agent delivery across the blood-brain barrier. AB - We have developed a brain tumor model in the nude rat utilizing NCI-N417D human small cell carcinoma of the lung grown both intracerebrally and s.c. The median latency period from the time of intracerebral tumor inoculation to the onset of neurological symptoms is 13 days with an intracerebral tumor take rate of 91% (29 of 32). The median survival is 13 days, and all animals were dead by Day 26. The tumor is discrete, well circumscribed, with occasional leptomeningeal spread and with minimal evidence of surrounding cerebral edema. Intracerebrally, this tumor is usually impermeable to Evan's blue:albumin (Mr 68,500) but not fluorescein (Mr 376). Although variable, the intracerebral tumor is less permeable to methotrexate than is the same tumor grown s.c. in the same animal (P less than 0.005). The intraarterial and i.v. routes of methotrexate administration in the presence and absence of blood-brain barrier opening were evaluated. Drug delivery to the intracerebral tumor and ipsilateral brain was significantly (P less than 0.025) greater when the methotrexate was given intraarterially and was significantly (P less than 0.0025) increased after osmotic blood-brain barrier opening. After barrier opening, methotrexate concentration was enhanced 3- to 4 fold in tumor and 10- to 20-fold in brain around tumor. Thus, the nude rat provides a model to investigate the biology and therapeutic responsiveness of human small cell carcinoma of the lung grown intracerebrally where it develops a blood-tumor barrier similar to that seen in humans. This model further provides the unique opportunity to investigate the role of osmotic blood-brain barrier opening in the treatment of a tumor which is sensitive to chemotherapeutic agents and for which tumor-specific monoclonal antibodies are available. PMID- 2985256 TI - Prognostic significance of laboratory parameters measured at diagnosis in small cell carcinoma of the lung. AB - An analysis of prognostic factors in small cell lung cancer has been carried out using data from 371 patients treated with identical chemotherapy in the context of a large prospectively randomized clinical trial. Prognosis was shown to be strongly correlated with initial performance status, disease extent, and routine biochemical tests at the time of diagnosis. Plasma albumin, plasma sodium, alkaline phosphatase, and gamma-glutamyl transpeptidase were all predictive of survival. An initial hemoglobin of less than 11 g/dl was also predictive, but age, sex, and initial WBC count were not. A multiple regression analysis identified performance status, plasma alkaline phosphatase, plasma sodium, disease extent, and plasma albumin as contributing independently to survival. Using these parameters, three prognostic groupings could be defined. The combination of performance status and the biochemical values more closely predicted survival than categorization on the basis of disease extent defined by clinical, radiological, and scanning criteria. Response to chemotherapy was strongly correlated with these prognostic groupings. A much higher response rate occurred in patients in the best prognostic category in whom the tumor mass is assumed to be smaller. These results provide a simple basis for predicting prognosis in small cell lung cancer and indicate that the better prognosis in chemotherapy responders is not solely due to the treatment. PMID- 2985257 TI - Establishment and identification of small cell lung cancer cell lines having classic and variant features. AB - Using a chemically defined medium containing hydrocortisone, insulin, transferrin, 17 beta-estradiol and selenium, with or without serum supplementation (2.5% v/v), continuous cell lines can be established from 72% of all fresh biopsy specimens of small cell lung cancer (SCLC) containing tumor cells. No differences were observed in the rate of establishing cell lines from newly diagnosed untreated patients, or from patients who have relapsed from prior therapy, or from a variety of different organ sites. Biochemical characterization of 50 SCLC cell lines for the expression of L-dopa decarboxylase; bombesin-like immunoreactivity; neuron-specific enolase, and the brain isozyme of creatine kinase, revealed that SCLC cell lines can be subdivided into two distinct classes: classic SCLC cell lines (35 lines), which express elevated levels of all four biomarkers; and variant SCLC cell lines (15 lines) which have undetectable levels of L-dopa-decarboxylase and bombesin-like immunoreactivity, but continue to express neuron-specific enolase and the brain isozyme of creatine kinase. The presence of the latter two markers distinguishes variant lines fron non-SCLC cell lines. In addition, four distinct classes were identified morphologically. The biomedical differences among established SCLC cell lines may account for the differences in response rates to cytotoxic therapy observed in newly diagnosed SCLC patients. A prospective study of biomarker characterization of SCLC tumors will determine if clinical differences exist between classic and variant SCLC tumors. PMID- 2985259 TI - Histomorphology of human lung cancer. AB - We have seen that neoplasms can arise at a number of sites through the lung. They arise from a variety of cells populating these sites. And, specific lesions of different cell types have a variety of biological behaviors. Some are very aggressive and spread rapidly throughout the body. Some are slower growing and are more amenable to therapy. PMID- 2985258 TI - Characterization of variant subclasses of cell lines derived from small cell lung cancer having distinctive biochemical, morphological, and growth properties. AB - We have described the establishment and biochemical characterization of 50 small cell lung carcinoma (SCLC) cell lines. Further analysis of these data, combined with studies of morphology and growth characteristics, indicates that 35 (70%) of the lines retained typical morphology (SCLC, intermediate subtype), growth characteristics (growth as tightly packed floating cellular aggregates, long doubling times and low colony-forming efficiencies), and biochemical profile (presence of L-dopa decarboxylase, bombesin-like immunoreactivity, neuron specific enolase, and high concentrations of brain isoenzyme of creatine kinase). They are referred to as classic SCLC lines. The remaining 15 (30%) lines had discordant expression of the biochemical markers; they retained high concentrations of brain isozyme of creatine kinase, but had significantly lower concentrations of neuron-specific enolase and lacked L-dopa decarboxylase and bombesin-like immunoreactivity. These cell lines are called variants. SCLC variant lines could further be divided into (a) biochemical variant lines having variant biochemical profile but retaining typical SCLC morphology and growth characteristics; and (b) morphological variant (SCLC-MV) lines having variant biochemical profile, altered morphology (features of large cell undifferentiated carcinoma) and altered growth characteristics (growth as loosely attached floating aggregates, relatively short doubling times and cloning efficiencies). Fifty-five clones derived from the three SCLC subclasses retained their parental phenotypes. In SCLC-MV lines there was a near constant relationship between variant morphology, altered growth characteristics and amplification of the c-myc oncogene; classic SCLC and biochemical variant SCLC lines were not amplified. Variant morphologies frequently are present in SCLC tumors at autopsy, and most SCLC-MV lines reflect changes that had occurred in the tumors from which they were derived. Because SCLC-MV tumors behave more virulently in the patient and are radioresistant in vitro, these findings are of considerable biological and clinical interest. PMID- 2985260 TI - Cellular mechanisms for tumor promotion and enhancement. AB - The concepts of tumor promotion discussed in this chapter demonstrate the complexity of the tumor promotion process, and illustrate the difficulty of determining which events are mechanistically important to promotion, and which ones are the result, or by-product of promotion. A number of possible mechanisms of promotion, both epigenetic and genetic, have been described on a molecular level. However, it should be stressed that none of these mechanisms is mutually exclusive; indeed, the enormous complexity of tumor promotion suggests that several of the mechanisms discussed above may very well be interrelated. The effects on epidermal differentiation, for example, may turn out to be the result of altered expression of particular oncogene whose product is actually a growth factor able to select for and amplify certain subpopulations of cells. Furthermore, the multiple steps which may occur at the molecular level, perhaps by way of sequential gene activation, serve to mirror the multiple stages which now delineate carcinogenesis in mouse skin. PMID- 2985261 TI - Tumor enhancement factors and mechanisms in the hamster respiratory tract carcinogenesis model. PMID- 2985262 TI - Cyclic AMP response to various haemopoietic regulators. AB - Diffusible inhibitors and stimulators are involved in the regulation of bone marrow pluripotent stem cell (CFU-S) proliferation. We have previously shown the existence of CFU-S inhibitors in foetal calf marrow and liver and have started their purification. The lack of a simple and time-saving test to determine the kinetic state of CFU-S and the activity of the inhibitors led us to explore the possibility of a biochemical proliferation marker that could be used for screening purpose. Since it was shown that cyclic AMP was implicated in the regulation of CFU-S proliferation, it was of interest to study the variations in cAMP levels after stimulation and inhibition of CFU-S entry into cycle. The results of in vitro experiments showed that the increase in cAMP levels observed in bone marrow cells after incubation with different haemopoietic stimulators was specific neither for bone marrow cells nor for the various haematopoietic regulators. In the in vivo experiments, an increased cAMP level was observed 8 hr after one injection of Ara-C at the time when CFU-S are recruited into S phase. However, no modification of cAMP levels has been observed after injection of CFU S inhibitors in the Ara-C-treated mice. Although cAMP does not seem to be a suitable marker for testing the activity of inhibitory fractions during the purification process, this work has contributed to the study of CFU-S stimulators. PMID- 2985263 TI - Increased gap junctional area in the rat liver after administration of dibutyryl cAMP. AB - Since cAMP has recently been reported to be a possible physiological modulator of cell-to-cell communication, we performed a quantitative freeze-fracture investigation on the hepatocyte gap junctions after administration of a membrane permeant derivative of this cyclic nucleotide. For this purpose, male rats received two intraperitoneal injections of 100 mg dibutyryl cAMP/kg body weight with a time interval of 2.5 h. Litter mates were injected with saline only. Five hours after the start of the treatment, tissue blocks of the left lateral liver lobe were fixed by immersion and processed for freeze-fracture. By point counting on negatives projected on a square double-lattice test system the relative gap junctional area on contiguous hepatocyte membranes was determined. As compared to control animals, the proportion of the membrane area occupied by gap junctions in dibutyryl cAMP-treated liver parenchyma significantly increased from 4.9% to 6.1%. Within the gap junctions no changes in shape, particle density or packing pattern were observed. Possibly, the enlarged gap junctional area provides structural pathways for the integration of the response of hepatocytes to messages mediated by cAMP. PMID- 2985264 TI - Platelet alpha-adrenergic receptor activity and pizotifen in migrainous children. AB - Adrenoceptors have been measured in the platelets from migrainous children, using (3H) yohimbine. Maximum binding (Bmax) was not significantly different in migrainous children and controls, 259 +/- 21 and 265 +/- 19 f moles per mg of protein respectively. Dissociation constants (KD) were significantly higher in the migraine group than in the controls 2.62 +/- 0.18 and 2.17 +/- 0.07 respectively (p less than 0.02). Bmax values were lower in 6 out of 9 cases following treatment with pizotifen. In 8 out of 9 cases KD values were lower following pizotifen treatment; in one case there was no change. PMID- 2985265 TI - A 43 kilobase cosmid P transposon rescues the fs(1)K10 morphogenetic locus and three adjacent Drosophila developmental mutants. AB - The K10 female sterility locus involved in establishment of the embryonic dorsoventral axis maps genetically to the 2E2-2F1 interval of the Drosophila X chromosome. We microdissected the 2E2-2F3 region from salivary gland chromosomes and used clones obtained from the microdissected fragments to establish a chromosomal walk covering more than 200 kb. To identify the K10 gene we used P mediated transformation with cosmid clones constructed in cos-P, a cosmid vector incorporating the terminal repeats of the P element. Clone cos9, containing a 43 kb insert, transformed the germ line of homozygous K10 females and allowed production of normal progeny. It also rescued three genes, crooked neck, pecanex, and kurz, which map genetically near K10. Transformation experiments using smaller fragments of cos9 localize the K10+ function within 11 kb. Northern blots hybridized with probes from this region indicate the presence of several mRNA species. Each transcript has been assigned to a complementation group. PMID- 2985266 TI - The Drosophila ninaE gene encodes an opsin. AB - The Drosophila ninaE gene was isolated by a multistep protocol on the basis of its homology to bovine opsin cDNA. The gene encodes the major visual pigment protein (opsin) contained in Drosophila photoreceptor cells R1-R6. The coding sequence is interrupted by four short introns. The positions of three introns are conserved with respect to positions in mammalian opsin genes. The nucleotide sequence has intermittent regions of homology to bovine opsin coding sequences. The deduced amino acid sequence reveals significant homology to vertebrate opsins; there is strong conservation of the retinal binding site and two other regions. The predicted protein secondary structure strikingly resembles that of mammalian opsins. We conclude the Drosophila and vertebrate opsin genes are derived from a common ancestor. PMID- 2985267 TI - Symbiotic mutants of Rhizobium meliloti that uncouple plant from bacterial differentiation. AB - Spontaneous mutants at a new symbiotic locus in Rhizobium meliloti SU47 are resistant to several phages and are conditionally insensitive to a monoclonal antibody to the bacterial surface, apparently because they are deficient in a wild-type exopolysaccharide. On alfalfa, the mutants do not curl root hairs, but penetrate the epidermis directly, forming nodules that contain no visible infection threads or "bacteroids," have a few bacteria in superficial intercellular spaces only and not within the nodule cells, and fail to fix nitrogen (Fix-). Evidently, infection threads are not essential for cell proliferation and nodule formation, which are here induced by a bacterial signal at a distance and uncoupled from the bacterial differentiation that normally goes on as well. PMID- 2985268 TI - The host protein required for in vitro replication of poliovirus is a protein kinase that phosphorylates eukaryotic initiation factor-2. AB - The HeLa cell protein (host factor) required for in vitro replication of poliovirus has been identified as a 67,000 dalton phosphoprotein. The purified protein displays three activities in vitro: stimulation of poliovirus RNA synthesis in the presence of poliovirus replicase, apparent self-phosphorylation, and phosphorylation of the alpha-subunit of eukaryotic protein synthesis initiation factor 2 (eIF-2). All three activities can be removed or inhibited by an antibody to host factor. Partially purified preparations of reticulocyte eIF-2 contain a similar phosphoprotein and display host factor activity in the viral RNA synthesis assay in vitro. In vitro phosphorylation of the 67 kd protein can be stimulated by low concentrations of double-stranded RNA. Addition of phosphorylated host factor in an in vitro RNA synthesis assay significantly changes the kinetics of viral RNA synthesis, indicating that protein phosphorylation may play an important role in viral RNA replication. PMID- 2985269 TI - Structure of the two distinct types of minichromosomes that are assembled on DNA injected in Xenopus oocytes. AB - DNA injected into germinal vesicles of Xenopus oocytes is assembled into two distinct types of minichromosomes. One type is soluble and behaves like conventional nucleosomal chromatin. The other type is insoluble, is sensitive to DNAase I and to micrococcal nuclease, lacks a canonical nucleosome repeat, and generates a half-nucleosome size limit digest with micrococcal nuclease. We suggest that these peculiar minichromosomes may be the ones that display the unconstrained, "dynamic" DNA supercoils in the living oocyte. PMID- 2985270 TI - Novel partitioning of DNA cleavage sites for Drosophila topoisomerase II. AB - We have examined the long-range distribution of double-stranded DNA cleavage sites for Drosophila melanogaster topoisomerase II. These studies reveal a novel partitioning of preferred topoisomerase II cleavage sites. In the eukaryotic DNAs examined, major cleavage sites were typically found in nontranscribed spacer segments and close to the 5' and 3' boundaries of genes. In contrast, there were few if any prominent cleavage sites within genes. In addition, most of the major topoisomerase II cleavage sites closely corresponded to naked DNA hypersensitive sites for the prokaryotic enzyme, micrococcal nuclease. PMID- 2985271 TI - Degradation of unassembled alpha- and beta-spectrin by distinct intracellular pathways: regulation of spectrin topogenesis by beta-spectrin degradation. AB - Analysis of the turnover of unassembled proteins during the assembly of the erythroid membrane skeleton has revealed that alpha- and beta-spectrin, two structurally related, high molecular weight proteins, are degraded in a selective manner by two distinct intracellular pathways. Unassembled alpha-spectrin (t1/2 approximately equal to 2 hr) is degraded by a system with all the pharmacological characteristics of a membrane-bound, lysosomal-type pathway. This result illustrates for the first time the selective degradation of an intracellular short-lived, unassembled protein by a lysosomal pathway. In contrast, unassembled beta-spectrin is degraded extremely rapidly (t1/2 approximately equal to 15-20 min at 38 degrees C) by a soluble cytoplasmic system in an apparently ATP independent manner. These observations suggest that the selective and rapid degradation of beta-spectrin serves an important regulatory role in the topogenesis of the spectrin-based membrane skeleton in the chicken erythrocyte. PMID- 2985272 TI - DNA-binding activity is associated with purified myb proteins from AMV and E26 viruses and is temperature-sensitive for E26 ts mutants. AB - Oncogene protein products from avian myeloblastosis virus, p48v-myb, and from avian leukemia virus E26, p135gag-myb-ets, are located predominantly in the nucleus of nonproducer bone marrow cell clones, as revealed by indirect immunofluorescence. Both oncogene proteins were purified by immunoaffinity chromatography using monoclonal antibodies against p19 and immunoglobulins specific for myb, which was expressed in bacteria for antibody production. The purified proteins bind to DNA in vitro. In contrast, purified p135gag-myb-ets proteins from several mutants of E26 virus, temperature-sensitive for myeloblast transformation, either lost their abilities to bind to DNA or exhibited highly thermolabile DNA-protein interactions in vitro. DNA binding of AMV and E26 oncogene proteins is inhibited by myb-specific immunoglobulins. Our results suggest that lesions in the myb oncogene affect transformation as well as DNA binding of myb proteins in vitro. PMID- 2985273 TI - EGF receptor affinity is regulated by intracellular calcium and protein kinase C. AB - Phorbol esters specifically reduce the binding of epidermal growth factor to surface receptors in intact cells, but not when added directly to isolated membranes. We show that after treatment of intact cells with phorbol myristate acetate, 125I-EGF binding is reduced in membranes prepared subsequently. High affinity binding of 125I-EGF is modulated by an intracellular calcium-dependent regulatory process. Preventing calcium entry with EGTA or enhancing intracellular calcium with A23187 in intact cells modulates EGF receptor affinity in membranes isolated subsequently. Also, EGTA attenuates the usual inhibition of EGF binding caused by phorbol esters. Membrane preparations do not respond to phorbol ester treatment because the calcium- and phospholipid-dependent protein kinase C is removed or inactivated during membrane isolation. Reconstitution of unresponsive membranes with purified C kinase alters phosphorylation of the EGF receptor and restores the inhibitory effect of phorbol esters on 125I-EGF binding previously observed only in intact cells. Thus, activation of the Ca++-dependent enzyme, C kinase, modulates EGF receptor affinity, possibly via altered receptor phosphorylation. PMID- 2985274 TI - Transgenic mice. PMID- 2985275 TI - Isolation of Z-DNA binding proteins from SV40 minichromosomes: evidence for binding to the viral control region. AB - Proteins dissociated from SV40 minichromosomes by increasing NaCl concentration were tested for their binding to Z-DNA [Br-poly(dG-dC)] and B-DNA [poly (dG-dC)]. Z-DNA binding proteins are largely released in 0.2 M NaCl whereas most B-DNA binding proteins are not released until 0.6 M NaCl. Incubation of SV40 minichromosomes with Z-DNA-Sephadex in low salt solution results in proteins with Z-DNA binding activity (PZ proteins). These proteins bind to negatively supercoiled DNAs containing left-handed Z-DNA but not to relaxed DNAs. They compete with anti-Z-DNA antibodies in binding to negatively supercoiled DNAs. The binding is tighter to negatively supercoiled SV40 DNA than to other plasmids, suggesting sequence-specific Z-DNA binding. PZ proteins binding to negatively supercoiled SV40 DNA interfere with cleavage at the Sph I sites, within the 72 bp repeat sequences of the viral control region, but not with cleavage at the Bgl I site, at the origin of replication. Removal of PZ proteins also exposes the Sph I sites in the SV40 minichromosomes while addition of PZ proteins makes the sites inaccessible. PMID- 2985276 TI - DNA intermediates and telomere addition during genome reorganization in Euplotes crassus. AB - Three gene-sized molecules cloned intact from macronuclear DNA served as hybridization probes to study excision of these molecules from chromosomes and their processing during macronuclear development in the hypotrich Euplotes crassus. These molecules occur in integrated forms within polytene chromosomal DNA during macronuclear developmental. After transection of the polytene chromosomes, the three molecules occur in intermediate forms. One of the three molecules first appeared in a large intermediate that was subsequently replaced by a second intermediate, approximately 140 bp larger than the final molecule. The other two macronuclear molecules were detected only in intermediates approximately 140 bp larger than the mature form. These penultimate intermediates are larger by virtue of oversized telomeres, which are pared to yield the mature gene-sized molecules. PMID- 2985277 TI - Forked, gypsys, and suppressors in Drosophila. AB - We have isolated DNA sequences defining the forked locus of Drosophila. All the alleles belonging to the right pseudoallelic series of M. M. Green map within a 5.4 kb DNA fragment that encodes four transcripts. These RNAs are only expressed in 2-4 day old pupae and their accumulation is affected by mutations at the forked locus. Also, homozygous mutations at the unlinked suppressor loci su(Hw) and su(f) cause the accumulation of these RNAs to return to the normal wild-type levels. The gypsy transposable element, which causes the mutant phenotype in the suppressible forked alleles, encodes a transcript that is maximally expressed in 2-3 day old pupae, the same time in development as transcription of the forked RNAs. We propose a model to explain the mutagenic effect of gypsy based on the influence of promoter elements in the long terminal repeats of the transposon on the transcription from other promoters located nearby. PMID- 2985278 TI - Greying with age in mice: relation to expression of murine leukemia viruses. AB - Some strains of C57BL/10 H-2-congenic mice were found to exhibit greying with age, whereas others did not. Two patterns of greying were observed, diffuse greying beginning at 4 to 6 months of age and patterned greying beginning at 4 to 6 weeks. Strains exhibiting either greying pattern expressed high levels of infectious ecotropic and mink cell focus-inducing murine leukemia viruses (MuLV) in tests of thymus and spleen and in cultures from skin or tail biopsies, whereas nongreying strains expressed little virus until late in life. Electron microscopy demonstrated large accumulations of MuLV in grey, but not in black areas, of skin from a mouse with patterned greying. Infectious MuLV was produced spontaneously by embryos of greying, but not of nongreying, mice and pups of nongreying strains fostered on greying mothers turned grey after 3 months. These results suggest that greying with age results from melanocyte dysfunction that occurs subsequent to pre- or early postnatal infection with MuLV. PMID- 2985279 TI - Delimitation and properties of DNA sequences required for the regulated expression of human interferon-beta gene. AB - The human interferon-beta (IFN-beta) gene contains sequences within its 5' flanking region that mediate the virus-induced activation of mRNA transcription. A series of mutant IFN-beta genes, including the 5' deletion mutants, was constructed and introduced into mouse L929 cells. Expression study of those mutant genes demonstrates that: sequences upstream, but not downstream, of -40 from the cap site are responsible for the viral induction of the gene; that the upstream boundary of the DNA sequences required to support the maximum level of induction lies between -117 and -105 from the cap site; and that this upstream sequence shows a property similar to enhancer elements as it functions in either orientation with a latitude in distance from the cap site. Within this sequence, we note the presence of repetitious hexanucleotides (consensus: A-A-AG-TG-G-A), each of which may play a role in the maximum induction of the IFN-beta gene. PMID- 2985280 TI - A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. AB - A strong transcription enhancer was identified in the genomic DNA (235 kb) of human cytomegalovirus (HCMV), a ubiquitous and severe pathogen of the herpesvirus group. Cotransfection of enhancerless SV40 DNA with randomly fragmented HCMV DNA yielded two SV40-HCMV recombinant viruses that had incorporated overlapping segments of HCMV DNA to substitute for the missing SV40 enhancer. Within HCMV, these enhancer sequences are located upstream of the transcription initiation site of the major immediate-early gene, between nucleotides -118 and -524. Deletion studies with the HCMV enhancer, which harbors a variety of repeated sequence motifs, show that different subsets of this enhancer can substitute for the SV40 enhancer. The HCMV enhancer, which seems to have little cell type or species preference, is severalfold more active than the SV40 enhancer. It is the strongest enhancer we have analyzed so far, a property that makes it a useful component of eukaryotic expression vectors. PMID- 2985281 TI - Cell-specific expression of a transfected human alpha 1-antitrypsin gene. AB - We have cloned the human alpha 1-antitrypsin (alpha 1-AT) gene and identified the promoter and the transcription initiation point. The cloned gene, following transfection, is expressed in a cell-specific manner, being transcribed in a human hepatoma cell line (Hep3B) but not in HeLa cells. We show that the 5' flanking region of the alpha 1-AT gene contains DNA sequences sufficient for efficient transcription in Hep3B but not in HeLa cells. This DNA sequence also activates, in a cell-specific manner, heterologous promoters such as that of SV40; however, the effect is only obtained in one orientation, suggesting that this cis-acting cell-specific element does not share all the features generally associated with enhancers. By cotransfection-competition experiments we also show the existence of a limiting trans-acting factor, essential for the expression of the alpha 1-AT gene in Hep3B cells. PMID- 2985282 TI - A high affinity topoisomerase I binding sequence is clustered at DNAase I hypersensitive sites in Tetrahymena R-chromatin. AB - Topoisomerase I is associated with DNAase I hypersensitive sites in the nontranscribed spacers flanking the rRNA genes in Tetrahymena thermophila. The endogenous topoisomerase I introduces site and strand specific single-strand cleavages in the rDNA spacers in situ. The cleavages occur base specifically within a hexadecameric sequence element present in two or three direct repeats at the hypersensitive sites. The sequence specificity and polarity of the cleavage reaction are identical when the enzyme is reacted with naked rDNA, indicating that the repetitive element functions as a high-affinity topoisomerase I attraction site in the r-chromatin. The biological mechanism associated with this phenomenon appears to be widespread among eukaryotes, since the topoisomerase I recognition sequence is conserved in the rDNA spacers of phylogenetically remote organisms, such as fungi, slime molds, ciliates, and insects. PMID- 2985283 TI - DNA topoisomerase II is required at the time of mitosis in yeast. AB - We have constructed five new temperature-sensitive DNA topoisomerase II mutations and have analyzed their physiological consequences in yeast. Several lines of evidence suggest that the activity of topoisomerase II is required specifically at the time of miosis. First, top2 mutations cause dramatic lethality at the restrictive temperature, but only if the mutant cells are actively traversing the cell cycle. Second, temperature-shift experiments with synchronized cultures show that the onset of inviability coincides with the time of mitosis. Third, fluorescence microscopy reveals that the normal progression of mitosis is disturbed in mutant cells at the restrictive temperature. Finally, inviability at the restrictive temperature is prevented by nocodazole, an inhibitor of tubulin polymerization that prevents formation of the mitotic spindle. These results are consistent with the hypothesis that the essential function of topoisomerase II is to allow the separation of intertwined chromosomal DNA molecules during mitosis. PMID- 2985284 TI - The termination region for SV40 DNA replication directs the mode of separation for the two sibling molecules. AB - Separation of the two newly replicated chromosomes in simian virus 40 late replicating intermediates (RI*) occurred by two pathways. The parental DNA strands were completely unwound, releasing circular DNA monomers with a gap in the nascent strand (Form II*), or duplex DNA in the termination region was not unwound, resulting in formation of catenated dimers. Under optimal conditions, both products were transient intermediates in replication, although Form II* was predominant. However, in hypertonic medium both RI* and catenated dimers accumulated, and Form II* was not observed. Hypertonic medium appeared to inhibit both DNA unwinding in the termination region and separation of catenated dimers. When the size of the genome or the position of the origin of replication was changed, termination occurred at sites other than that of wild-type SV40. Neither catenated dimers nor RI* DNA accumulated at these sites. Instead, RI* separated into Form II*. Unwinding parental DNA was more difficult at some termination regions than others. Therefore, although completion of DNA replication does not require a unique termination sequence, this sequence can determine the mode of separation for sibling molecules. PMID- 2985285 TI - A bidirectional rho-independent transcription terminator between the E. coli tonB gene and an opposing gene. AB - We identified an Escherichia coli gene, designated P14, that is adjacent to and in the opposite orientation to the tonB gene. The 36 base pair intercistronic region between tonB and P14 contains a novel rho-independent transcription terminator that functions bidirectionally, both in vivo and in vitro, to terminate tonB and P14 transcription. Transcription of tonB and P14 terminates at symmetrically equivalent nucleotides, such that the 3' ends of tonB and P14 transcripts are complementary. The terminator is 70% efficient in both directions in vitro. Interestingly, relative rates of in vivo RNA synthesis, immediately prior to and following the terminator, appear to indicate that it is more efficient in the tonB direction (95%) than in the P14 direction (70%). We discuss the possibility that this gene arrangement has regulatory consequences for the expression of tonB. PMID- 2985286 TI - Extended life span and tumorigenicity of nonestablished mouse connective tissue cells transformed by the fos oncogene of FBR-MuSV. AB - We have analyzed the transforming potential of two fos oncogene products in nonestablished cultures of mouse connective tissue cells: p55fos of FBJ-MuSV and p75gag-fos of FBR-MuSV. Although both proteins induced morphological transformation and colony formation at low cell density in a G418 resistance selection assay, p75gag-fos exhibited more pronounced transforming potential than p55fos. In addition, p75gag-fos-transformed cells overcame crisis with a high probability and were tumorigenic in syngenic mice. These properties of the FBR MuSV appear to be linked to structural alterations in the p75gag-fos oncogene product. Polyoma virus large T protein complemented the transforming potential of fos, in that it not only increased the probability of establishment of fos transformed cells but also enhanced fos-induced morphological transformation. Our results suggest that different oncogenes affect morphological transformation, low cell density growth, establishment, and tumorigenicity to various degrees. PMID- 2985287 TI - cAMP induces a transient elevation of cGMP levels during early culmination of Dictyostelium minutum. AB - Oscillatory cAMP signals very likely organize the cell movement which leads to fruiting body construction in Dictyostelium minutum [Schaap, P., Konijn, T.M. and Van Haastert, P.J.M.: Proc. Natl. Acad. Sci. USA 81, 2122-2126 (1984)]. Stimulation with cAMP induces a transient elevation of cGMP in cells at the early culmination stage, which peaks at 12-18 s. A half maximal cGMP response is induced by 10(-7) M cAMP and saturation of the response is reached at 10(-5) M cAMP. No cGMP accumulation was induced by stimulation of vegetative or aggregative cells of D. minutum by cAMP. Since the transient increase of cGMP is most likely involved in the transduction of chemotactic signals, our results indicate that cAMP signals organize fruiting body formation by inducing chemotaxis inside the aggregate. PMID- 2985288 TI - The interaction of resealed reticulocyte ghosts with Fe3+-transferrin-CO3(2-). AB - The present studies were designed to investigate the interaction of Fe3+transferrin-CO3(2-) with the transferrin receptors of the resealed reticulocyte ghost and to assess the degree to which the iron release reaction can be reconstituted in resealed ghosts supplemented with entrapped cytoplasmic components. Reticulocyte, but not erythrocyte, ghosts displayed an intact Fe3+transferrin-CO3(2-) binding capability. When ATP, NADH and ferritin were included during the resealing process, some iron release to the ghosts was observed. PMID- 2985289 TI - [Poliovirus type 2 in sewage canals in Prague in 1981-1982]. PMID- 2985290 TI - [Relation among different types of rectal cancer classified by traditional Chinese medicine from the observations on immunological function]. PMID- 2985291 TI - [Determination of cyclic nucleotide and sialic acid in patients with symptoms of blood stasis and its value in assessing the therapeutic effect of drugs for activating blood circulation and removing stasis]. PMID- 2985292 TI - In vitro transformation of rat pleural mesothelial cells by chrysotile fibres and/or benzo[a]pyrene. AB - Rat pleural mesothelial cells (RPMC) were treated, in vitro, in a two-stage model of carcinogenesis. RPMC obtained from passage 12th were treated once with 1 microgram/ml of benzo-[a]pyrene (BP) and from passage 13th to 40th with 0.4 microgram/cm2 of chrysotile fibres (Chr). Transformation was determined by the observation of the colonies formed when the cells were plated at low density. Colonies were classified into four classes according to the level of lack in contact inhibition between cells (0 to III). BP as well as chrysotile fibres were potent in inducing a high proportion of type III colonies which were not observed in control series. In addition, there was no synergistic effect between BP and Chr. These results would indicate that chrysotile fibres do not act as a promoter on rat pleural mesothelial cells in culture, but induce morphologically transformed colonies. PMID- 2985293 TI - Impaired inotropic responses to alpha-adrenergic stimulation in experimental left ventricular hypertrophy. AB - We have previously reported that left ventricular hypertrophy in two-kidney, one clip renal hypertensive rats (2K-1C RHRs) was associated with diminished inotropic responsiveness to isoproterenol and glucagon, suggesting an alteration in the receptor-adenylate cyclase cascade. The present study was performed to investigate the hypothesis that in these same hearts, inotropic responses to alpha-adrenergic stimuli could be enhanced as a compensatory mechanism. alpha Adrenergic stimulation was achieved by graded phenylephrine infusion (1.02 to 41.2 microM/min) in the presence of propranolol (10(-7) M). The inotropic response was evaluated in the isovolumetric isolated rat heart (Langendorff preparation) paced at 260 beats/min. Results showed a significantly reduced inotropic response to alpha 1-adrenergic stimulation in 2K-1C RHR hearts irrespective of perfusion pressure (50 or 80 mm Hg [PP50 or PP80]) (+427.5 +/- 62.1 vs +1236 +/- 216.4 mm Hg X sec-1 at PP50, p less than .01 and +339 +/- 98.3 vs +1440 +/- 254 mg Hg X sec-1 at PP80, p less than .001) even when comparison was made at equivalent myocardial flow rates (RHR hearts perfused at 80 mm Hg vs control hearts perfused at 50 mm Hg). Quantitative assessment of number of alpha 1-adrenergic receptors (3H-prazosin binding) showed a significant decrease compared with that in age-matched sham-operated normotensive control rats (45 +/- 2.5 vs 64 +/- 1.7 fmol/mg protein, p less than .001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985294 TI - Digoxin immunoreactivity in cord and maternal serum and placental extracts. Partial characterization of immunoreactive substances by high-performance liquid chromatography and inhibition of Na+, K+-ATPase. AB - Digoxin-like immunoreactive substances (DLIS) have been successfully extracted and concentrated from cord serum, mixed (cord and maternal) serum and placentas. Similar substances have also been extracted from normal adult serum, but DLIS in this medium are present in much lower concentrations. Concentrated DLIS have been separated into several immunoreactive fractions by use of reverse-phase high performance liquid chromatography. Immunoreactive fractions were tested for their ability to inhibit the Na+,K+-ATPase by measuring the 86Rb-uptake of red blood cells in the presence of these fractions. A potent inhibitor was identified in an immunoreactive fraction which also contains progesterone, but the results suggest that the pump inhibitor is not progesterone. Cross-reactivity studies utilising fluorescence polarization immunoassay have shown that cortisone is the most potent immunoreactive substance of cord serum. PMID- 2985295 TI - Serum ACTH levels in patients with polycystic ovarian disease. AB - Serum adrenocorticotrophic hormone levels were assayed in 8 patients with polycystic ovaries and a similar number of control subjects. Serum ACTH concentrations were normal in 6 patients and marginally elevated in 1. Investigations of markedly elevated readings in another patient showed this to be due to an interfering factor rather than the ACTH molecule itself. PMID- 2985296 TI - Postcoital test after vaginal washing with NaHCO3. AB - The Authors treated with vaginal washing of NaHCO3 (Sodium Bicarbonate) 55 patients whose postcoital tests were persistently poor or negative. The cervical pH increase made the test positive in about 30% of cases and three pregnancies occurred. This treatment with NaHCO3 proved to be useful, handy and welcome to patients. PMID- 2985297 TI - Copper and iron complexes catalytic for oxygen radical reactions in sweat from human athletes. AB - Sweat collected from 'explosive' and 'endurance' athletes immediately after exercise contains low molecular mass complexes of copper detectable by their ability to bind to o-phenanthroline. Concentrations of these copper complexes are much greater in arm sweat than in trunk sweat. The iron content of arm sweat, as determined by the ferrozine method, is also greater than that of trunk sweat. However, much of the iron in trunk sweat exists in a low molecular mass form that can bind to the antibiotic bleomycin, whereas the iron in arm sweat does not exist in this form. The metal complexes in human sweat are capable of stimulating the peroxidation of membrane lipids in the presence of ascorbic acid. The physiological significance of the presence of iron and copper complexes in sweat is discussed. PMID- 2985298 TI - Serum collagenase-like peptidase activity in rheumatoid arthritis and systemic lupus erythematosus. AB - Collagenase-like (CL) peptidase activity in serum, which was measured using a newly synthesized substrate, (succinyl-Gly-Pro-Leu-Gly-Pro)-4-methylcoumaryl-7 amide, was significantly lower in patients with advanced rheumatoid arthritis or with systemic lupus erythematosus than that in normal controls. Decrease of the serum enzyme activity was more pronounced in systemic lupus erythematosus. No significant change in serum CL-peptidase activity was found in other connective tissue diseases such as mixed connective tissue disease and Sjogren's syndrome. PMID- 2985299 TI - Phosphatidic acid phosphatase activity in subcellular fractions of normal and dystrophic human muscle. AB - Biopsy samples from normal and dystrophic human muscle (Duchenne type) were fractionated by differential centrifugation and microsomes, mitochondria and cytosol were assayed for phosphatidic acid phosphatase (EC 3.1.3.4) and marker enzymes of mitochondria and cytosol. The activity of phosphatidic acid phosphatase was significantly lower in microsomes and higher in cytosol and mitochondria of dystrophic muscle than in the corresponding subcellular fractions of normal muscle. The results support an explanation of earlier findings that there is reduced G3P incorporation into diglycerides and phosphatidylcholine and a qualitative and quantitative change in the amount of phosphatidylcholine in dystrophic microsomes. The possible reasons for the reduction in the activity of only microsomal PA-P-ase were discussed. PMID- 2985300 TI - Pituitary Cushing's disease arising from a previously non-functional corticotrophic chromophobe adenoma. AB - Severe pituitary Cushing's disease of sudden onset after 18 years of unsuccessful treatment for a previously non-functioning chromophobe adenoma is described in a middle-aged woman. Initial presentation with symptoms of optic nerve compression had been preceded by two years of amenorrhoea. Transfrontal resection of a chromophobe adenoma followed by radiotherapy, performed both at this time and again after a recurrence eight years later, failed to eradicate the tumour. Ten more years elapsed before she rapidly developed florid features of Cushing's syndrome. Plasma ACTH levels were markedly elevated and were only partially reduced by further transfrontal surgery, complete removal of the tumour proving impossible. Subsequent bilateral adrenalectomy was performed to control her hypercortisolism and a course of cytotoxic chemotherapy was administered in an attempt to treat the tumour recurrence. Immunocytochemical staining of tumour obtained at surgery demonstrated ACTH immunoreactivity both before and after the development of Cushing's disease. Although silent corticotrophic adenomas of the human pituitary, including chromophobic tumours, have been previously described, this is the first report of such a tumour becoming functional, sufficient to cause clinically evident Cushing's disease. PMID- 2985302 TI - A case of pituitary dependent Cushing's disease with clinical and biochemical features of the ectopic ACTH syndrome. AB - A case of atypical pituitary dependent Cushing's disease is reported. The patient presented with clinical symptoms similar to those of the ectopic ACTH syndrome; notably a marked hypokalaemic alkalosis, widely fluctuating plasma cortisol levels, greatly elevated plasma ACTH levels, and failure to suppress both plasma cortisol and ACTH levels following high dose oral dexamethasone. However, a large aggressive pituitary tumour was detected by skull X-ray and computed tomography. Removal of the pituitary tumour led to full remission of the patient's Cushing's syndrome. Pro-opiomelanocortin (POMC) related peptides in the plasma and tumour tissue extract of this patient have been characterized by gel-filtration and Concanavalin-A Sepharose affinity chromatography, indicating processing of POMC in a manner more usually associated with ectopic tumours. PMID- 2985301 TI - Endocrine effects of low dose aminoglutethimide as an aromatase inhibitor in the treatment of breast cancer. AB - The endocrine effects of low dose (62.5 mg, twice a day) aminoglutethimide (AG), without hydrocortisone (HC), escalating at monthly intervals to a conventional dose of AG (500 mg twice a day) combined with HC, were studied in 33 postmenopausal breast cancer patients. Pretreatment serum concentrations of oestrone (E1) and oestradiol (E2) were significantly suppressed by 62.5 mg of AG twice a day. Although further suppression of E1 appeared to occur with 125 mg of AG twice a day, this was not statistically significant. For E1 and E2, higher doses of AG or combined AG and HC failed to cause further significant suppression compared with that obtained at 125 mg of AG twice a day. Significant rises in serum androstenedione were found with all doses of AG alone, although pretreatment concentrations of androstenedione were not significantly altered by combined AG and HC treatment. Mean pretreatment concentrations of dehydroepiandrosterone sulphate (DHA-S) were significantly suppressed by 62.5 mg of AG twice a day and further marked suppression occurred on combined AG and HC therapy. Serum cortisol, aldosterone and plasma ACTH concentrations showed no significant alterations throughout treatment. Aminoglutethimide is as effective at 125 mg twice a day without HC in its suppression of oestrogen levels as at 500 mg twice a day with HC, and its use in this form warrants clinical evaluation. PMID- 2985303 TI - Iopanoic acid is of minimal benefit in the treatment of severe hyperthyroidism: conclusions from a case study. AB - Iopanoic acid (1 g/d) was used together with propylthiouracil (1200 mg/d) in the treatment of a patient with very severe hyperthyroidism and associated cardiac failure. Although serum total T3 decreased by 75% within 48 h and reached normal after 72 h, free T3 levels did not fall to normal. Total and free T4 remained markedly elevated and features of hyperthyroidism persisted. Estimations of theoretical in vivo occupancy of nuclear thyroid hormone receptors, based on serum free T4 and free T3, suggest that the marked decrease in total T3 would not result in a corresponding decrease in thyroid hormone action. Hence, estimates of potential benefit from oral cholecystographic contrast agents, based only on measurements of total T3, may be unduly optimistic. When temporary agranulocytosis developed in this patient, the prior use of iopanoic acid, by markedly reducing thyroidal iodine uptake, restricted the therapeutic options. Caution should, therefore, be exercised in the use of iodine-containing contrast media as adjunctive antithyroid agents. PMID- 2985304 TI - Immunoprecipitation of TSH-TSH receptor complexes. AB - The ability of Graves' sera to interact with the TSH receptor crosslinked to a 125I-labelled photoactive derivative of TSH has been investigated. Crosslinked complexes were prepared using non-purified detergent solubilized human thyroid and guinea pig fat TSH receptors. Affinity purified porcine TSH receptor preparations wee also used. After crosslinking, the crosslinked TSH-TSH receptor complexes were separated from aggregates and free TSH on Sephacryl S-300, incubated with test sera followed by immunoprecipitation using anti-IgG or Protein A. Using non-purified human TSH receptors crosslinked to TSH, a mean +/- SD of 12.1 +/- 4.9% of the crosslinked complex was immunoprecipitated with Graves' sera (n = 7) compared with 10.3 +/- 2.6% with Hashimoto sera (n = 6; P greater than 0.14) and 3.8 +/- 1.0% with normal sera (n = 6; P less than 0.004). These values were markedly reduced when TSH receptor preparations free of other thyroid autoantigens (guinea pig fat TSH receptors) were used. Under these conditions immunoprecipitation with Graves' sera (n = 24) was 1.6 +/- 1.3% compared with 0.8 +/- 0.6% for Hashimoto sera (n = 13) and 0.8 +/- 0.4% for normal sera (n = 12; P less than 0.003). In addition complexes formed between TSH and affinity purified porcine TSH receptors gave low immunoprecipitation values for Graves' (1.44 +/- 0.73%; n = 20) and Hashimoto sera (1.7 +/- 0.94; n = 11) which were not significantly different (P greater than 0.4). Overall, therefore, the effects of Graves' and Hashimoto sera were similar and the amounts of material immunoprecipitated were markedly reduced when TSH receptor preparations containing reduced amounts of other autoantigens were used. Consequently the Graves' sera did not appear to interact specifically with crosslinked TSH-TSH receptor complexes. However the Graves' sera studied did contain TSH receptor antibodies which could inhibit the binding of labelled TSH to TSH receptors in the preparations used and our results suggest that the binding of TSH and these antibodies to the receptor is mutually exclusive. There is considerable evidence that serum from patients with Graves' disease contains antibodies to the TSH receptor (Rees Smith, 1981). Several studies have suggested that binding of the receptor antibody and TSH to the TSH receptor is mutually exclusive (Manley et al., 1977; Petersen et al., 1977; Rickards et al., 1981) but recently the formation of termolecular complexes consisting of detergent solubilized receptors, labelled TSH and Graves' IgG has been reported (Konishi et al., 1982; De Bruin et al., 1984).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985305 TI - Lymphoid cells produce an immunoregulatory glucocorticoid increasing factor (GIF) acting through the pituitary gland. AB - Products are released in vitro by mitogen stimulation of human peripheral blood mononuclear cells or rat spleen cells which increase corticosterone blood levels when injected into normal rats. We report that an almost pure population of human peripheral blood lymphocytes in mixed lymphocyte culture stimulated with phytohaemagglutinin also produces a glucocorticoid increasing factor(s). The product equivalent to the amount released by 5 X 10(5) cells increased corticosterone levels four-fold and also increased ACTH levels. When rats were hypophysectomized or treated with dexamethasone to block ACTH output, no corticosterone increase was noted following administration of glucocorticoid increasing factor(s) (GIF). Similar results were obtained with supernatants of rat spleen cells stimulated with concanavalin A. We conclude that GIF has no direct action on the adrenals in vivo and that a functional pituitary gland is essential for its action. The presented data taken together with those described earlier suggest the existence of a glucocorticoid associated immunoregulatory feedback circuit. PMID- 2985306 TI - Biochemical and physiochemical characteristics of a pre-B cell stimulating factor produced by the plasmacytoma cell line LICR LON HMY2. AB - We have characterized the pre-B cell colony stimulating activity (pre-B cell CSA) from LICR LON HMY2 conditioned medium (CM) by a variety of biochemical techniques. Pre-B cell CSA was found to be associated with a heat stable glycoprotein which has an isoelectric point of 8.3 and a mol. wt, as determined by polyacrylamide gel electrophoresis, of 28-32 kD. The relationship of this activity to previously described factors acting on cells of the B cell lineage is discussed. PMID- 2985308 TI - Treatment of chronic renal failure with dietary fiber. PMID- 2985307 TI - Maintenance of cytomegalovirus (CMV) latency and host immune responses of long term renal allograft survivors. I. Prolonged suppression of in vitro lymphocyte responses against CMV infected fibroblasts related to previous secondary CMV infection. AB - Cytomegalovirus (CMV) specific humoral and cellular immunity was investigated in 16 renal allograft recipients with long term graft survival (26-122 months) who were shown to be CMV seropositive before transplantation. Results were compared with healthy individuals with latent CMV infections. Recipients (n = 9) who experienced a symptomatic secondary CMV infection shortly after transplantation (less than 6 months), showed a prolonged but finally temporary suppression of their in vitro lymphocyte memory responses against CMV infected fibroblasts (CMV FF; median SI: 1.9), a persistence of high antibody titres against intracellular CMV antigens and most of them also had antibodies against CMV membrane antigens (CMV MA). In contrast the recipients (n = 7) who could maintain their CMV in latency after transplantation, had lower antibody titres and their in vitro memory lymphocyte responses against CMV-FF (median SI: 9.3) were comparable to those of the healthy controls (median SI: 11.6). The memory lymphocyte responses against purified CMV virions were depressed in both recipient groups. These results suggest that cellular immunity against CMV infected target cells constitute an important mechanism in maintaining CMV in latency after allografting. PMID- 2985309 TI - Activity of helix: destabilizing protein in lymphocytes of patients with glomerulopathies and with chronic renal insufficiency. AB - The aim of this work was to determine the activity of helix-destabilizing protein (HD-protein), which is indispensable in basic genetic processes such as DNA replication, genetic recombination and transcription, in lymphocytes of patients with different morphological types of glomerulonephritis and with chronic renal insufficiency, treated conservatively and by maintenance hemodialysis. The estimation was done on 44 patients with glomerulonephritis and on 20 patients with chronic renal insufficiency. The control group consisted of 20 healthy individuals. The investigations prove the fact that in human lymphocytes helix destabilizing protein occurs, and its quantity enables us to determine its activity in the diseased and healthy state. Measurement of HD-protein activity seems to be a good marker of the altered lymphocyte function. Especially high HD protein activity was noticed in cases of acute glomerulonephritis, lupus glomerulonephritis and in the active forms of chronic glomerulopathies. Generally, the tested parameter showed a positive correlation with the activity of the glomerulonephritic process. In lymphocytes of patients with chronic renal insufficiency treated conservatively, a greater activity of helix-destabilizing protein occurred than in the control group. Lymphocytes from patients with chronic renal insufficiency, treated with repetitive maintenance hemodialysis, did not reveal significant differences in HD-protein activity, determined directly before hemodialysis and after its completion, in comparison to the control group of healthy individuals. PMID- 2985311 TI - Elevated CK-MB and CK-BB in serum and tumor homogenate of a patient with lung cancer. AB - Elevated serum levels of creatine kinase (CK) were found in a patient with small (oat) cell carcinoma of the lung. Fractionation of the enzyme showed markedly elevated CK-MB and CK-BB isoenzymes. Clinical and subsequent pathological examination showed no evidence of infarction, inflammation, or tumor involvement of the heart; however, analysis of tumor tissue for CK showed predominance of CK MB and CK-BB isoenzymes, thus implicating tumor as the source of the circulating levels of CK-MB and CK-BB. Our case is the first to document CK-MB from neoplastic tissue homogenates, and illustrates that markedly elevated circulating levels of CK-MB, or increased levels of CK-MB in combination with CK-BB may point away from a myocardial source, and toward the existence of a malignancy. PMID- 2985310 TI - Auto-oxidation and a membrane-associated 'Fenton reagent': a possible explanation for development of membrane lesions in sickle erythrocytes. AB - Sickle erythrocytes spontaneously generate approximately twice-normal amounts of activated oxygen species (superoxide, peroxide and the highly reactive hydroxyl radical). In addition, they contain excessive amounts of membrane-bound haemichromes, which presumably accumulate because of some degree of sickle haemoglobin instability, perhaps involving interactions between haemoglobin and the lipid bilayer. Since most erythrocyte antioxidant mechanisms are located in the cytoplasm, it is hypothesized that a physiologically significant hydroxyl radical generator would be located within or directly adjacent to the membrane. Indeed, sickle erythrocyte membranes appear to contain a biological 'Fenton reagent', the iron of which is able to cycle between ferrous and ferric states and thereby facilitate superoxide/peroxide-driven hydroxyl radical generation. Data suggest that at least some haemichromes could fulfil this function, although many other potential hydroxyl radical generators exist in sickle erythrocytes. A particularly interesting - but unproven - possibility is membrane-associated haem (without globin). It is further hypothesized that the various membrane abnormalities of sickle erythrocytes might be caused by excessive auto-oxidation. Although proof of this hypothesis does not yet exist, a number of observations are consistent with it. Certainly, evidence indicates that some oxidative modification of sickle erythrocyte membrane proteins and lipids has taken place. Few data exist regarding the state of the erythrocyte membrane in the unstable haemoglobinopathies, but evidence of oxidative perturbation of the HbKoln erythrocyte membrane has been reported. Of great interest is that the membranes of thalassaemic and sickle erythrocytes appear to be remarkably similar in terms of possible oxidative phenomena, suggesting that the membranes of these two cell types may be subjected to similar oxidative stresses. PMID- 2985313 TI - Herpes genitalis: clinical features and treatment. PMID- 2985312 TI - Familial hypocalciuric hypercalcemia: description of a new kindred with emphasis on its difference from primary hyperparathyroidism. AB - In order to better understand the difference between familial hypocalciuric hypercalcemia (FHH) and primary hyperparathyroidism (PHP), 4 adults with this disease (FHH(+], from the same kindred, and spanning 2 generations, were compared with 6 patients with PHP, 10 normal controls (N) and 3 unaffected members of the same kindred (FHH(-]. Clinically speaking, the FHH(+) patients were asymptomatic while those with PHP had recurring renal stones. Various phosphocalcic balance elements were measured in blood and urine, in a fasting state, following oral loading of 1 g calcium, as well as in 24-hour urine specimens. Serum calcium levels were slightly higher (11.90 +/- .14 vs 11.37 +/- .24 mg/dl; p less than .05), in patients with FHH(+), but serum phosphorus (2.47 +/- .38 vs 2.88 +/- .40 mg/dl), serum parathyroid hormone (301 +/- 147 vs 291 +/- 224 pg/ml) and serum 1,25(OH)2D (52.8 +/- 6.7 vs 55.0 +/- 12.0 pg/ml) values were similar to those observed in patients with PHP. Urinary calcium present in FHH(+), while fasting (.064 +/- .043 mg/100 ml G.F.), after the calcium loading (.150 +/- .071) and in the 24-hour specimen (.089 +/- .016), was below the 95% confidence limit of values obtained in patients with PHP (.225 +/- .026, p less than .001; .413 +/- .066, p less than .001; 314 +/- .080, p less than .001), but similar to those obtained in N and FHH(-).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985314 TI - Herpes simplex virus vaccines--where are we? PMID- 2985315 TI - Bovine brain gangliosides in the treatment of human sensory and motor neuronal degenerations. AB - Report of an open trial on the effect of bovine brain gangliosides on patients with sensorimotor neuronal degeneration. The results were encouraging and stress the need for well controlled trials. PMID- 2985317 TI - Incidental finding of intense thyroid radiogallium activity during febrile illness. AB - Two elderly adults presented with fever of undetermined origin. Radiogallium imaging showed intense concentration in the thyroid, without abnormal localization elsewhere. In the first case, a repeat radiogallium study performed three weeks later was within normal limits. The patient had 0% uptake of radioiodide, and no demonstrable iodine within the thyroid. This probably represented thyroiditis. In the second case, an elevated T4 (RIA) and borderline elevated T3 resin uptake suggested the "dumping phase" of thyroiditis. The two cases indicate that "benign" diseases of the thyroid can be associated with diffuse radiogallium concentration in the gland. PMID- 2985316 TI - Importance of markers in Tc-99m pyrophosphate myocardial images. AB - A false-positive Tc-99m PYP myocardial scan may result due to gastric activity from breakdown of radiopharmaceutical and accumulation of Tc-99m pertechnetate in gastric cells. Importance of anatomic landmarks during early blood pool images is emphasized to prevent this error. PMID- 2985318 TI - Scintigraphic characterization of amyloid cardiomyopathy. AB - Amyloidosis is an important entity in the differential diagnosis of cardiac failure of undetermined etiology. In this case report, the typical pattern of combined systolic and diastolic impairment in amyloid cardiomyopathy was demonstrated by analysis of the cardiac blood pool study. In addition, the patient described had mild uptake of Ga-67 citrate, as well as the characteristically intense myocardial uptake of Tc-99m pyrophosphate. Scintigraphic assessment may be particularly helpful when the diagnosis of amyloidosis is being considered in a patient with unexplained cardiac failure. PMID- 2985319 TI - Early detection of anthracycline-induced cardiotoxicity by stress radionuclide cineangiography in conjunction with Fourier amplitude and phase analysis. AB - Thirty-three cases of anthracycline related cardiotoxicity occurred in our institution in patients with a previously negative cardiovascular history, physical examination, and normal ECG. A total of 95 RNCA studies were performed in this group (73 studies included both rest and exercise RNCA). Twenty-one patients had two or more serial studies. Seventeen had a normal, baseline prechemotherapy study and 16 had studies done following the initiation of therapy. Fourier analysis, consisting of amplitude and phase images, were created for each study. In the subset with a baseline study, the rest LVEF became abnormal first in two of 17 patients (12%), the rest or stress LVEF in ten of 17 (59%), the rest Fourier image analysis in ten of 17 (59%), and the rest or stress Fourier image analysis in 16 of 17 (94%). In the subset without a baseline study, the rest LVEF was abnormal in ten of 16 (63%), the rest or stress LVEF in 15 of 16, (94%), the rest Fourier image analysis in 16 of 16 (100%), and the rest or stress Fourier image analysis in 16 of 16 (100%). The authors conclude that: 1) the exercise RNCA is superior to the rest RNCA alone in the early detection of anthracycline related cardiotoxicity, 2) the single most sensitive indicator of cardiotoxicity is Fourier image analysis; and 3) sequential rest and stress RNCA studies with Fourier amplitude and phase analysis is the most sensitive, noninvasive method of evaluating patients who receive potentially cardiotoxic agents. PMID- 2985321 TI - The diagnosis of thyroid disease on bone scans. AB - Three cases are reported in which visualization of the thyroid occurred during Tc 99m pyrophosphate bone scans. All were found to be hyperthyroid with elevated serum thyroid hormone and two patients also had elevated 4-hour radioactive iodine uptakes. PMID- 2985322 TI - Selective, symmetric, skeletal muscle uptake of Tc-99m pyrophosphate in rhabdomyolysis. PMID- 2985320 TI - A comparison of the tumor-seeking agent Tc-99m(V) dimercaptosuccinic acid and the renal imaging agent Tc-99m dimercaptosuccinic acid in humans. AB - Being aware of the ideal nuclear properties of Tc-99m, our interest has been focused on the design of the (+5) oxidation state Tc-99m(V) dimercaptosuccinic acid (Tc(V)-DMSA) as a tumor-seeking agent. Tc-99m(V) DMSA holds a TcO4(3-) core and, like PO4(3-), has excellent characteristics for tumor uptake, but has a different distribution than the well-known renal scanning agent, Tc-99m DMSA. The differences in chemical behavior of Tc-99m(V) DMSA and Tc-99m DMSA are discussed. Three cases in which neoplasms were studies with Tc-99m(V) DMSA and Tc-99m DMSA are presented. Tc-99m DMSA and Tc-99m(V) DMSA, having a common ligand and tracer but, with the metal ion core in a different oxidation state, the uptake characteristics are altered markedly. PMID- 2985323 TI - Carcinoma of the stomach with osseous and extra-osseous metastases. Visualization of primary and secondary sites with Tc-99m medronate. PMID- 2985324 TI - Effect of clavulanic acid on inactivated amoxicillin excretion in urinary tract infection. AB - Oral amoxicillin was taken with and without clavulanic acid by normal subjects and by patients with chronic complicated urinary tract infection to examine the in vitro protective effect of clavulanic acid on amoxicillin degradation. When amoxicillin alone was taken, urinary excretion of the penicilloic acid of amoxicillin in bacteria-positive patients was higher than that in bacteria negative patients and in normal subjects. There was no comparable change in urinary penicilloic acid excretion in the presence of clavulanic acid. There were significant in vitro protective effects of clavulanic acid on beta-lactamases in the urine. PMID- 2985326 TI - Treatment of cancer-associated hypercalcemia with mithramycin and oral etidronate disodium. PMID- 2985325 TI - Measurement of proteoglycans, elastase, collagenase and protein in synovial fluid in inflammatory and degenerative arthropathies. AB - The number of leucocytes and the concentrations of protein, proteoglycans (PG), elastase a1 proteinase inhibitor complexes (E-alpha 1 Pi) and collagenolytic activity were measured in the synovial fluid (SF) of 15 patients with rheumatoid arthritis (RA) and 18 with osteoarthritis (OA). The mean levels of protein, E alpha 1 Pi and collagenase and the number of leucocytes were higher in RA than in OA SF. However, the mean level of PG was higher of OA SF than in RA. In the latter, they were principally in the form of monomers and fragments while in the former they were in the form of aggregates and monomers. There was a direct relationship between the concentration of E-alpha 1 Pi and either the number of white cells or the concentration of synovial proteins, suggesting that the measurement of E-alpha 1 Pi complexes is a biochemical index of the local inflammatory reaction. There was an inverse correlation between the concentrations of PG and E-alpha 1 Pi which may reflect the effect of degradation in PG of elastase and other enzymes released at the same time. Finally, there was a direct relationship between the concentration of E-alpha 1Pi and collagenase which may be the reflection of a simultaneous release of various enzymes from leucocytes and macrophages. PMID- 2985327 TI - Computed tomographic demonstration of obstruction of the inferior vena cava caused by hepatocellular carcinoma. AB - A case of hepatocellular carcinoma obstructing the hepatic portion of the inferior vena cava by extrinsic compression demonstrated on computerized tomography is presented. To our knowledge, only one previous case has been reported which demonstrates actual invasion of the inferior vena cava by hepatic tumor. The signs of obstruction include: nonvisualization of the intrahepatic portion of the IVC, dilatation of the infrahepatic IVC with paravertebral venous collaterals and lack of changes in the diameter of the IVC after Valsalva maneuver. PMID- 2985328 TI - Computed tomographic detection of gas within spontaneously necrotic tissue. AB - Computed tomography (CT) is a widely accepted modality for the detection, localization, and potential therapeutic drainage of abdominal abscesses. The CT demonstration of extraluminal gas within the abdomen usually signifies the presence of an abscess. We report two cases of nonpurulent spontaneous tissue necrosis which simulated a gas containing abscess on CT. The significance of a gas-containing mass detected by CT must be interpreted cautiously since this appearance is not necessarily pathognomonic of a drainable abscess. Confirmation of purulent material within a suspected abscess collection requires documentation by percutaneous aspiration or surgery. PMID- 2985329 TI - Gestational trophoblastic neoplasia in southern Connecticut: the experience of the Yale Trophoblastic Center, 1971-1983. PMID- 2985330 TI - Serum stimulation of lysyl hydroxylase activity in cultured human skin fibroblasts. AB - In the absence of ascorbic acid, confluent human skin fibroblasts incubated in 0.5% serum-supplemented medium had one-third of the level of lysyl hydroxylase activity of cells incubated in media containing high serum concentrations (5 20%). This difference appeared to be due to a decline in the enzyme activity following serum deficiency, and was largely abolished by addition of ascorbic acid to the medium. The effect of serum deficiency was slow, manifesting in 48 h at the earliest, and was completely reversed by replenishing the medium with serum. Prolyl hydroxylase activity was independent of serum concentration, both in the absence and in the presence of ascorbic acid in the culture medium. PMID- 2985331 TI - The effects of the GnRH agonist, [(imBzl)-D-His6, Pro9-NEt]-GnRH, on the response to stress in rats. AB - The antigonadal activity of potent gonadotropin-releasing hormone (GnRH) analogs may be associated with undesirable secondary effects on other aspects of the endocrine system. In preliminary studies, rats treated with GnRH agonists were noted to have altered adrenal weights. To determine whether this was associated with alterations in the pituitary-adrenal axis we have studied male and female rats treated with the GnRH agonist [(imBzl)-D-His6, Pro9-NEt]-GnRH (GnRH-A). Animals receiving daily doses of this peptide showed normal adrenal corticosteroid responsivity to both ether and immobilization stress. Brain, pituitary, and plasma concentrations of immunoreactive (IR) beta-endorphin and ACTH were unaffected in treated animals after four weeks of daily injections, although after one week plasma levels of the two hormones were transiently elevated in female rats. Adrenal, thyroid, and pituitary lobe weights were unchanged with treatment, except for an increase in anterior pituitary weight in males receiving the lowest dose of GnRH-A. In conclusion, long-term treatment with GnRH-A, while significantly affecting gonadal and secondary sexual tissue, had little impact on the hypothalamo-pituitary-adrenal system or on stress responsivity in rats. PMID- 2985332 TI - Cutaneous histiocytoma in a goat. AB - A cutaneous histiocytoma involving the scrotum was diagnosed in a 14-month-old French Alpine buck. A wedge biopsy was taken for histologic examination. The remaining tumor regressed spontaneously and had completely resolved within six months of initial observation. PMID- 2985333 TI - Distinct rotaviruses isolated from asymptomatic calves. AB - Rotaviruses were isolated following cell culture of the intestinal contents of four non-diarrheic calves. The four isolates were serially propagated in MDBK and BSC-1 cells in the presence of trypsin and produced rotavirus particles morphologically similar to those found associated with diarrhea. They were antigenically related to the Nebraska calf rotavirus (Norden strain) as investigated by immunofluorescence. Three isolates could be distinguished from the reference Nebraska rotavirus by their thermal stability and/or their differential responses to intestinal neutralizing antibodies. Two isolates produced on BSC-1 cells plaques significantly different in size from the reference strain, No significant genomic variations were detected among the isolates. PMID- 2985335 TI - Computed tomography of renal masses. AB - Computed tomography (CT) has provided a valuable imaging modality for the study of renal masses. Simple cysts are the most common of renal masses, and they are usually readily differentiated by CT from the wide variety of solid tumors that affect the kidneys. This article describes the CT technique which is used for evaluation of renal masses; the CT appearances of such renal tumors as renal cell carcinoma, renal pelvic carcinoma, lymphoma, oncocytoma, and angiomyolipoma and the use of CT in the staging of malignant renal tumors. The CT findings in various renal cystic diseases are also described. The role of CT for evaluating renal masses in relation to such other imaging techniques as sonography and angiography is discussed. PMID- 2985334 TI - Nutrition and metabolism of the geriatric dog. AB - Sixteen 10-12-year-old and eight 1-year-old dogs were studied over a two year period to determine comparative differences in physiological response to 4 diets varying in protein content and percentages of energy contributed by protein. The ability of old dogs to utilize nutrients as supplied by these foods was not significantly different from that of young adult dogs. Except for indices of mitogenic stimulation and serum urea nitrogen (SUN) other physiological parameters studied were not affected by the diet fed. Regardless of diet, old dogs had significantly higher serum levels of cholesterol, phosphorus and alkaline phosphatase and had lower indices of mitogenic stimulation than did young dogs. PMID- 2985336 TI - Clinical evaluation of computed tomography images in esophageal cancer after submucosal injection of iodized oil. AB - A submucosal injection of iodized oil (Lipiodol) was administered to patients with esophageal cancer and computed tomography scanning was performed. Infiltration to the surrounding organs, especially the aorta, vertebra, trachea, bronchi, heart, and lymph nodes was examined and the results compared with those from plain computed tomography. Compared with plain computed tomography findings, invasion to the submucosa, adventitia, and surrounding organs was much more clearly demonstrated. This method accurately delineated the oral side of mucosal lesions, which could be useful in establishing a plan for radiotherapy. PMID- 2985337 TI - Hepatic excretion of urographic contrast medium: use in delineating intrahepatic cholangiocarcinoma on scans delayed for 2 hours. AB - Vicarious excretion of urographic contrast media by the liver has been detected by conventional radiographic methods in patients with renal impairment or unilateral obstruction. It occurs in patients with normal renal function, however, and can be detected by delayed postcontrast computed tomography scans. A case is presented in which an intrahepatic cholangiocarcinoma was best depicted on scans delayed for 2 hours. After 2 hours, the contrast medium remained within hepatic parenchyma but was virtually cleared from the vascular and interstitial spaces. PMID- 2985338 TI - Studies on HSV specific IgA antibodies in lacrimal fluid from patients with herpes keratitis by solid phase radioimmunoassay. AB - A highly sensitive solid phase radioimmunoassay (RIA) was adapted for determination of herpes simplex virus (HSV) specific IgA antibodies in lacrimal fluids. The RIA procedure was applied to examine HSV IgA antibodies in serial samples of lacrimal fluid and in sera from 14 patients with herpes keratitis. HSV specific IgA antibodies were detected in the lacrimal fluid of the affected eye in 12 of the 14 patients. HSV IgA antibodies were also detected in the lacrimal fluid of the unaffected eye of 7 of these patients in titers lower than those found in the corresponding affected eye. Evidence for local HSV antibody production in lacrimal fluid in herpes keratitis was supported by examination for antibodies to the unrelated measles antigen. Detection of lacrimal fluid HSV IgA antibodies might be a useful adjunct in rapid diagnosis of herpes keratitis. The advantage of characterizing the local immune response as well as viral expression in the affected vs. the unaffected eye in the same individual as a model for understanding the factors leading to subclinical vs. clinical manifestations of recurrent ocular herpes virus infections will be discussed. PMID- 2985339 TI - Na+-K+ ATPase distribution in frog and bovine lenses. AB - Na+-K+ ATPase activity was measured in the capsule-epithelium and decapsulated frog and bovine lens. The decapsulated lens contained approximately 20% of the whole lens activity in the frog and 30% in the bovine. These values were measured from the aqueous homogenate of the entire decapsulated lens, an approach which may have underestimated the activity by diluting the ouabain-sensitive component in the preparation. Subsequent determinations were done on separate portions of superficial (2 to 3 mm) anterior-equatorial, and posterior bovine cortex. The activities per gram of tissue were enriched with respect to the values for the entire decapsulated bovine lens. These activities were further enriched by a sucrose density gradient centrifugation. The anterior-equatorial cortical segment contained 1.6 times the activity found in the capsule-epithelium. The posterior cortex had a much smaller but statistically significant level of Na+-K+ ATPase. It is unlikely that the observed asymmetry of the anterior-equatorial segment with the posterior cortex is exclusively due to epithelial contamination for the result would require the adherence of 62% of the epithelium. Scanning electron micrographs of 6 decapsulated bovine lenses indicated an average contamination of about 9%. This asymmetry may have a physiological role in assisting the pump mechanism of the epithelium. PMID- 2985340 TI - Lung cancer and pleural effusion. Clinical significance and study of pleural metastatic locations. AB - We reviewed results of 78 diagnostic thoracoscopic examinations of patients with lung cancer and homolateral pleural effusion. The study stresses the main locations of pleural metastasis in cases of lung cancer and correlates these data with the pleural spread from extrathoracic primary neoplasms. Also pointed out is the operability of the present series and the different behavior in pleural metastatic spread according to the histologic type with correlation between gross appearance of the effusion and the cytology. PMID- 2985341 TI - Acyclic pyrimidine nucleoside analogues: influence on growth of L5178y mouse lymphoma cells and antiherpes activity in KB cells. AB - Several uracil and cytosine nucleoside analogues with 2-hydroxyethoxymethyl, 2 aminoethoxymethyl or 1,3-dihydroxypropoxymethyl side chains were synthesized and evaluated for cytostatic (L5178y mouse lymphoma cells) and antiviral (herpes simplex virus type 1 infected KB cells) activity. Two compounds exhibited antiherpesvirus activity. These were 1-(2'-hydroxyethoxymethyl)-5-aza-cytosine and 1-(1',3'-dihydroxypropoxymethyl)-5-iodouracil. The MIC values were 25.8 and 325.7 micrograms/ml, respectively. PMID- 2985342 TI - [Malignant tumors of the large vessels. 2 case reports]. PMID- 2985343 TI - [Endoscopic gluing of postoperative bronchial fistulas]. PMID- 2985344 TI - The mechanism and pattern of banding induced by restriction endonucleases in human chromosomes. AB - The mechanism of chromosome banding induced by restriction endonucleases was analyzed by measuring the amount of radioactivity extracted from [14C]thymidine labeled chromosomes digested first with restriction enzymes and subsequently with proteinase K and DNase I. Restriction enzymes with a high frequency of recognition sites in the DNA produced a large number short DNA fragments, which were extracted from chromosomes during incubation with the enzyme. This loss of DNA resulted in decreased chromosomal staining, which did not occur in regions resistant to restriction enzyme digestion and thus led to banding. Subsequent digestion of chromosomes with proteinase K produced a further loss of DNA, which probably corresponded to long fragments retained in the chromosome by the proteins of fixed chromatin. Restriction enzymes induce chromatin digestion and banding in G1 and metaphase chromosomes, and they induce digestion and the appearance of chromocenters in interphase nuclei. This suggests that the spatial organization and folding of the chromatin fibril plays little or no role in the mechanism of chromosome banding. It was confirmed that the pattern of chromosome banding induced by AluI, MboI, HaeIII, DdeI, RsaI, and HinfI is characteristic for each endonuclease. Moreover, several restriction banding polymorphisms that were not found by conventional C-banding were detected, indicating that there may be a range of variability in the frequency and distribution of restriction sites in homologous chromosome regions. PMID- 2985345 TI - [Physiological regulation and clinical significance of 18-hydroxy-11 deoxycorticosterone]. PMID- 2985346 TI - [Electron microscopy manifestations of pituitary adenoma with acromegaly and gigantism]. PMID- 2985347 TI - Influences of age on the positive inotropic effect mediated by alpha- and beta adrenoceptors in rat ventricular strips. AB - The positive inotropic effect of phenylephrine (in the presence of atenolol) and isoprenaline was studied in isolated right ventricular strips obtained from Wistar rats aged 2 weeks, 3 months, and 26 months. The positive inotropic effects induced by phenylephrine and isoprenaline were higher in the 2-week-old than in the 3- or 26-month-old rats. The typical 'alpha-type' or 'beta-type' responses were induced by phenylephrine (plus atenolol) or isoprenaline, in all the age groups. The EC50 values for the alpha-mediated positive inotropic effect decreased with age, while the EC50 value for the positive inotropic effect of isoprenaline increased in the senescent heart. These results suggest an enhanced alpha-adrenergic sensitivity in the senescent heart. PMID- 2985348 TI - [Ultrastructural organization of the neuromuscular synapses in red muscle fibers of rats during increased motor activity]. PMID- 2985350 TI - [Inverted interconnected base sequences in the Alu, HindIII and KpnI structure of DNA repetitive sequences]. PMID- 2985349 TI - [Interaction of oxygen anion radicals with eye melanins and ommochromes]. PMID- 2985352 TI - [Guar gum in the treatment of diabetes and hypercholesterolemia]. PMID- 2985351 TI - Haemodynamic, hormonal, and diuretic effects of felodipine in healthy normotensive volunteers. AB - Felodipine and placebo were infused in a double-blind, crossover study in 10 healthy normotensive volunteers. Compared with placebo, felodipine caused a significant decrease in diastolic blood pressure and forearm vascular resistance, while there was no change in systolic blood pressure. The rises in heart rate, plasma renin activity and plasma noradrenaline (norepinephrine) concentration further demonstrated the vasodilating activity of felodipine. Plasma aldosterone, adrenaline (epinephrine) and antidiuretic hormone concentrations were similar after a 90-minute infusion of felodipine or placebo. The response of plasma aldosterone levels to exogenous adenocorticotrophic hormone showed evidence of a slight blunting during felodipine infusion. Felodipine had a marked diuretic effect, probably secondary to an increase in natriuresis, which might be due to a direct tubular effect of the drug. PMID- 2985353 TI - Differing effects of low and high bulk maternal dietary supplements during pregnancy. AB - The study attempted to determine if the bulk of dietary supplements given to pregnant women after midgestation affects fetal growth. 127 Zulu women were randomly assigned to four groups, two of which received daily food supplements designed to raise their energy, protein and vitamin intakes to levels recommended by the U.S. Food and Nutrition Board. One of these supplements had a high bulk, the other a low bulk. The third supplement contained only zinc and the fourth a placebo. Women in all four groups had a similar weight gain and length of gestation. Birth weights were from 6.5 to 9.5% greater with the low bulk than with the other supplements. PMID- 2985355 TI - Re-evaluation of "frontal deafferentation" studies on the regulation of ACTH secretion in the light of CRF immunohistology. AB - Frontal afferents to the medial basal hypothalamus of the rat were interrupted by a Halasz knife, and 4 weeks later the brains were processed for immunostaining of CRF-fibers. It was found that such intervention saves most of the CRF immunoreactive fibers of the paraventriculo-infundibular tract. Some of the conflicting results of earlier deafferentation studies are discussed on the basis of CRF immunohistology. PMID- 2985356 TI - Lack of preventive effect of 6-hydroxy-dopamine on a decrease of beta-adrenergic receptors induced by repeated stress in the heart of rats. AB - Immobilization stress (2.5 h daily) repeated daily for 7 days resulted in a significant decrease in the number of beta-adrenergic receptors in heart compared to control values. Administration of 6-hydroxy-dopamine failed to prevent the stress-induced reduction in the number of beta-adrenergic receptors in the heart of rats as compared to intact stressed rats. These results support the concept that beta-adrenergic receptors in the heart of rats are primarily under the control of the noradrenergic nervous system. PMID- 2985354 TI - Changes induced in rabbit plasma and aqueous humour by ACTH4-10 and ACTH1-24. AB - In the rabbit the effect of a single i.m. dose of ACTH4-10 (130 micrograms kg-1) was compared with that of ACTH1-24 (20 micrograms kg-1). In contrast to ACTH1-24 the heptapeptide ACTH4-10 did not stimulate the adrenal secretion of corticosterone. Both ACTH fragments induced a slight hyperglycemia and in the aqueous humour they increased the concentration of proteins. PMID- 2985357 TI - Extracts and auto-oxidized constituents of certain plants inhibit the receptor binding and the biological activity of Graves' immunoglobulins. AB - Freeze-dried extracts (FDE) of the plants Lycopus virginicus, Lycopus europaeus, Melissa officinalis, and Lithospermum officinale, as well as products of the oxidation of certain of their constituents, have been shown to exert antithyrotropic activity by virtue of their ability to form adducts with TSH that bind weakly, if at all, to the TSH receptor. The thyroid-stimulating immunoglobulin G (IgG) found in the blood of patients with Graves' disease (Graves'-IgG) resemble TSH in their ability to bind to the thyroid plasma membrane, probably at the TSH receptor, and to activate the gland. In view of this similarity, and since some of the aforementioned FDE have been used in the treatment of hyperthyroidism in Graves' disease, we undertook studies of the effect of these FDE and their constituents on the binding and biological action of Graves'-IgG. In all samples of Graves'-IgG tested, incubation with antithyrotropic FDE or their antithyrotropic auto-oxidized constituents decreased their TSH-binding inhibitory activity in a dose-dependent manner. FDE from L. officinale also inhibited in a dose-dependent manner the direct binding to human thyroid membranes of a 125I-labeled preparation of receptor-purified Graves'-IgG. As judged from both stimulation of adenylate cyclase activity in vitro (thyroid stimulating Ig activity) and enhancement of thyroid iodine release in the McKenzie assay system (LATS activity), antithyrotropic FDE and their auto oxidized constituents also inhibited the biological responses to Graves'-IgG. FDE and constituents lacking antithyrotropic activity had little or no effect on the TSH-binding inhibitory activity, thyroid-stimulating Ig activity, or LATS activities of Graves'-IgG. Evidence of some degree of specificity of the inhibitory effects of the active compounds on Graves'-IgG was obtained in the demonstration that they failed to inhibit both the direct binding of [125I]insulin to its receptors in human lymphoblastoid IM-9 cells and the ability of IgG preparations containing antiinsulin receptor antibodies to inhibit the binding of labeled insulin. These observations suggest that the active principles in those FDE and their oxidized constituents with antithyrotropic activity may interact with the pathogenically important components of Graves'-IgG to inhibit their ability to bind to the TSH receptor and activate the thyroid, as they do with TSH. Our findings provide a possible rationale for the empirical, though poorly documented, use of FDE in the treatment of Graves' disease and some support for the suggestion that Graves'-specific IgG may have structural similarities to TSH itself. PMID- 2985358 TI - Prostaglandin E2-induced luteinizing hormone-releasing hormone release involves mobilization of intracellular Ca+2. AB - The present in vitro experiments were performed to examine the involvement of Ca+2 in the mechanism by which prostaglandin E2 (PGE2) induces LHRH release from the median eminence (ME) of the hypothalamus. Stepwise decreases in the Ca+2 concentration of the incubation medium reduced the LHRH response to PGE2. Nevertheless, neither complete omission of Ca+2 (residual Ca+2 concentration, 3.5 microM) nor chelation of residual Ca+2 with EGTA prevented the stimulatory effect of the PG, suggesting that a significant portion of the PGE2 effect on LHRH release is independent of extracellular Ca+2. Blockade of Ca+2 influx with verapamil, a Ca+2 entry blocker, demonstrated that this component of the PGE2 effect is completely independent of inward Ca+2 movement. Depletion of intraterminal Ca+2 stores by exposure of the MEs to the Ca+2 ionophore A23187 in medium without Ca+2 containing EGTA almost completely obliterated the subsequent LHRH response to PGE2, indicating that normal intraterminal Ca+2 levels are important for the PGE2 effect to occur. Preloading the ME terminals with 45Ca+2 and subsequent stimulation with PGE2 demonstrated that even in the absence of extracellular Ca+2, PGE2 stimulates Ca+2 efflux from the terminals, and this Ca+2 movement occurs temporarily associated with LHRH release. Depolarization of ME terminals with 56 mM K+ in the presence of normal Ca+2 concentration resulted in massive efflux of 45Ca+2 and a greater LHRH response than that produced by PGE2, suggesting that the effect of PGE2 is not the consequence of a nonspecific general depolarization of ME nerve terminals. Thus, although a full LHRH response to (exogenous) PGE2 necessitates normal extraterminal Ca+2 concentrations, the results indicate that translocation of Ca+2 from intracellular stores is an event involved in the mechanism by which PGE2 releases LHRH. PMID- 2985359 TI - Estrogen dependence of luteinizing hormone receptor expression in cultured rat granulosa cells. Inhibition of granulosa cell development by the antiestrogens tamoxifen and keoxifene. AB - In rat ovarian granulosa cells cultured for 48 h, addition of 10(-8) M estradiol (E2) enhanced choleragen-induced cAMP formation and LH receptor content by 2-fold and 6-fold, respectively. Two potent antiestrogens, tamoxifen and keoxifene, inhibited these effects of E2 in a concentration-dependent manner and significantly reduced cAMP production and LH receptors below the levels induced by choleragen. Both antiestrogens (greater than or equal to 1 microM) also reduced the effects of choleragen on cAMP levels and LH receptor content in the absence of exogenous E2. In addition, the antiestrogens (1 microM) inhibited the stimulatory effects of FSH and forskolin on granulosa cell maturation, as well as the enhancement of their actions by exogenous E2. FSH caused a concentration dependent rise in endogenous E2 accumulation during the 48-h culture period, suggesting that antiestrogens may prevent FSH-stimulated increases in LH receptors by inhibiting the actions of newly formed E2. Tamoxifen prevented the induction of LH receptors by 8-bromo-cAMP, indicating that its effects were on both cAMP production and cAMP action, whereas keoxifene predominantly altered granulosa cell development by its inhibition of estrogen effects on cAMP production. Although both exogenous E2 and the antiestrogens modified cAMP accumulation and LH receptor expression largely during the second 24 h of culture, their actions commenced during the first day. The antiestrogens had no effect alone and did not reduce the DNA content of granulosa cells. Also, they could be washed from the cells after 48 h of culture with complete recovery of forskolin-stimulated cAMP responsiveness by 72-96 h of culture. At a lower concentration (0.4 microM), tamoxifen, but not keoxifene, acted as a partial estrogen agonist since it enhanced choleragen action. These results indicate that the cAMP-mediated induction of LH receptors in cultured granulosa cells is dependent upon the continued actions of estrogen throughout the maturation process. PMID- 2985360 TI - Differential action of decidual luteotropin on luteal and follicular production of testosterone and estradiol. AB - Decidual tissue of the rat produces a hormone with physiological and biochemical characteristics similar to those of PRL. Because PRL affects both follicular and luteal production of testosterone and estradiol, it was of interest to determine whether decidual luteotropin affects basal and/or LH-stimulated ovarian secretion of steroids and whether it differentially affects follicular and luteal synthesis of testosterone and estradiol. The uteri of pseudopregnant adult rats were scratched on day 5 to induce decidual tissue formation. Pseudopregnant animals without decidua were used as controls. Rats were either hypophysectomized on day 8 or left intact. They were treated with 1.5 IU hCG/day or with vehicle between days 8-9. On day 9, blood was obtained from the ovarian vein, and both corpora lutea and large antral follicles were isolated and incubated in vitro. The presence of the decidua significantly suppressed both basal and hCG-stimulated ovarian secretion of estradiol, yet enhanced progesterone production. A similar inhibitory effect of decidual tissue on hCG stimulation of testosterone and estradiol was observed in the hypophysectomized rats. When the effect of decidua on follicles and corpora lutea was studied separately, it was found that follicles of rats with decidua produced significantly less testosterone and estradiol than follicles of rats without decidua. hCG administration to either intact or hypophysectomized rats markedly enhanced the follicular capacity to produce these two steroids. However, the degree of hCG stimulation of follicular steroidogenesis was significantly reduced by the presence of decidual tissue. In contrast, the decidua did not inhibit the in vitro steroidogenic capacity of corpora lutea. Luteal tissue of intact rats with or without decidua produced similar basal amounts of testosterone and estradiol and responded to a hCG challenge with comparable increases in the production of both steroids. After hypophysectomy, however, the responsiveness of corpora lutea to hCG stimulation differed in rats with or without decidual tissue. Whereas luteal cells of rats without decidual tissue gradually lost their responsiveness to hCG stimulation, luteal cells of rats with decidua remained highly responsive to hCG and produced high levels of testosterone and estradiol. In summary, the present investigation demonstrates that decidual luteotropin impairs ovarian secretion of estradiol and significantly inhibits the stimulatory effect of hCG on ovarian secretion of testosterone and estradiol.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985361 TI - Demonstration of a 60K molecular weight luteinizing hormone-releasing hormone receptor in solubilized adrenal membranes by a ligand-immunoblotting technique. AB - A new technique for the identification of LHRH receptors has been developed and applied to demonstrate an adrenal LHRH binding protein. Solubilized membrane proteins were separated electrophoretically and transferred to nitrocellulose paper. This was followed by sequential incubations with LHRH, anti-LHRH antiserum, peroxidase-conjugated second antibody, and 4-chloro-1-naphthol. Using an antiserum directed towards the middle region of LHRH, a 60K mol wt band was visualized in rat adrenal and pituitary membranes. A band of slightly higher molecular weight was present in membranes of bovine adrenal cortex but was absent in the medulla. The 60K band was not visualized when nonimmune rabbit serum was used. The 60K band was also not visualized when an antiserum requiring the NH2 and COOH termini of LHRH was used, suggesting that these regions of LHRH are not accessible to the antiserum after binding to the receptor. These studies have demonstrated the existence of LHRH binding protein in adrenal cortical tissue with a molecular size similar to that of the pituitary receptor. Adrenal membrane binding sites were less clearly demonstrated by conventional 125I-ligand binding techniques as nonspecific binding was high. The ligand-immunoblotting technique is a sensitive, specific and rapid procedure with potential application in screening normal and tumor tissues for LHRH receptors and studying LHRH interactions with its receptor. PMID- 2985362 TI - Turnover of growth hormone receptors in rat adipocytes. AB - Adipocytes isolated from the epididymal fat pads of normal rats specifically bound [125I]human GH [( 125I]hGH). Preincubation of cells with 20 micrograms/ml cycloheximide, an inhibitor of protein synthesis, produced a progressive loss of ability to bind [125I]hGH specifically. Loss of binding sites with time followed first order kinetics and had a half-time of about 45 min regardless of whether GH was present or absent during treatment with cycloheximide. Nonspecific binding of labeled hormone was unchanged by cycloheximide. Similar results were obtained when adipocytes were incubated with 200 micrograms/ml puromycin, another inhibitor of translation, but incubation with 5 micrograms/ml actinomycin D, an inhibitor of transcription, for 2.5 h had no effect on the binding of [125I]hGH by adipocytes. The findings are not attributable to cell death, since oxidation of [U-14C] glucose to 14CO2 and binding of [125I]insulin were unaffected in replicate cell populations exposed to the same treatments. Diminished binding could not be attributed to an effect of cycloheximide to hasten the degradation of receptor-bound hGH. Treatment of adipocytes with 0.1 mg/ml trypsin for 10 min virtually abolished their ability to bind [125I]hGH specifically, but binding capability gradually returned after removal of trypsin and was nearly restored to pretrypsin levels by 2 h. Addition of cycloheximide to the incubation medium after removal of trypsin completely prevented recovery of binding capability. Covalent binding of [125I]hGH to its receptors with disuccinimidyl suberate followed by sodium dodecyl sulfate-gel electrophoresis and autoradiography of proteins isolated from adipocyte membranes revealed three specifically labeled bands corresponding to mol wt of 250-300, 130, and 56 kilodaltons. Treatment of adipocytes with cycloheximide before cross-linking resulted in a proportional reduction in all three labeled bands, suggesting a similar half-life for all three entities. Similarly, all three labeled entities reappeared in parallel as adipocytes recovered from treatment with trypsin. The data strongly suggest that receptors for GH turn over rapidly on the surface of adipocytes and that ongoing protein synthesis is required to maintain binding capacity. The data do not permit distinction between rapid turnover of the receptor proteins themselves and a short-lived protein(s) which might be required to insert the receptors into the membrane. PMID- 2985363 TI - Release of luteinizing hormone and follicle-stimulating hormone from pituitaries of lactating rats by gonadotropin-releasing hormone and high potassium concentration. AB - Anterior pituitaries (APs) from lactating rats were studied in vitro to determine the mechanisms by which LH release, but not FSH release, is suppressed. Hemipituitaries were exposed for 4 h to GnRH (20 nM) or elevated potassium (K+; 57 mM) according to two different regimens: 6 min/h (pulsatile) or 60 min/h (continuous). Pulsatile K+ exposure stimulated similar amounts of LH release by APs from diestrous females and lactating females on day 5 or 10 who were nursing two or eight pups. Pulsatile GnRH stimulation resulted in significantly greater amounts of LH release from all groups than was observed after pulsatile K+, except by APs from females nursing eight pups. In this group, GnRH was no more effective than K+ in stimulating LH release by APs from day 5 of lactation, but was slightly but significantly more effective on APs from day 10 of lactation. The degree of suppression of GnRH-stimulated LH release was directly correlated with the intensity of the suckling stimulus, and removal of the eight-pup suckling stimulus for 48 h completely restored the LH response to GnRH to the response observed by APs from females on diestrous day 2. Exposure of the APs continuously to either K+ or GnRH produced results that were similar to those observed after the pulsatile regimen, except that the total amount of LH released during the 4-h incubation was greater. However, APs from females suckling eight pups still released significantly less LH in response to GnRH than did APs from females suckling two pups or diestrous females. FSH secretion by the APs was similar in response to either pulsatile K+ or GnRH stimulation and was not suppressed by suckling of a large litter. The total amount of FSH released by pituitaries from lactating and cycling animals did not differ significantly, except that by APs females deprived of their eight-pup litters for 48 h, in which case pulsatile GnRH stimulated a greater amount of FSH release. However, the pattern of FSH release varied, in that pulsatile GnRH stimulation of diestrous pituitaries showed a priming response, whereas no priming response was evident during lactation. Exposure of APs to elevated K+ continuously produced little more FSH release than exposure to pulsatile K+ stimulation. However, continuous stimulation by GnRH generally released more FSH than did K+. FSH responses to GnRH by pituitaries from lactating females were similar or greater than the FSH responses by pituitaries from diestrous females.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985364 TI - Assay of insulin mediator activity with soluble pyruvate dehydrogenase phosphatase. AB - The putative mediator of intracellular insulin action has been assayed quantitatively by its ability to increase the activity of solubilized pyruvate dehydrogenase (PDH) phosphatase. Conversion of soluble beef heart PDH b to PDH a by PDH phosphatase increased when incubation was carried out in the presence of a crude insulin mediator fraction generated from insulin-treated adipose tissue or liver plasma membranes. Increased PDH phosphatase activity was proportional to the concentration of added insulin mediator. Mediator generation was rapid, with a half-time of approximately 45 sec and was insulin dose dependent. Half-maximal mediator activity was produced at 0.3 nM added insulin, with maximal activity being generated at approximately 3 nM insulin. Mediator activity was significantly decreased at 7 nM insulin, but was increased 4-fold after ethanol extraction. Mediator behaved as an activator of PDH phosphatase, apparently by abolishing the inhibitory effects of ATP on phosphatase activity, but had no effect on PDH kinase activity. The assay of insulin mediator activity described here can be carried out under standardized conditions, in contrast to previously described methods using particulate mitochondrial preparations. PMID- 2985365 TI - The influence of temperature on the functions of cultured Sertoli cells. AB - Three functions of Sertoli cells were examined with cells from rats aged 13 and 25 days at 34 C, 38 C, and 40 C, namely protein synthesis (incorporation of amino acids into material precipitated by trichloroacetic acid), transport of alpha aminoisobutyric acid, and production of lactate. A fourth function, namely production of cAMP was examined at two temperatures (34 C and 38 C) in cells from rats aged 25 days. In all cases, activity was greater at 38 C than at 34 C and greater at 40 C than at 38 C. Lysate of Sertoli cells showed similar differences in protein synthesis at the three temperatures, again at both ages. Protein synthesis was higher at 38 C than at 34 C in Sertoli cell-enriched tubules from rats aged 25 days and in Sertoli cells from normal rats of the same age cultured on rat tail collagen. The difference in protein synthesis at the two temperatures was seen whether the cells were cultured, before the experiment, for 7 days at 38 C or at 34 C. Production of cAMP by Sertoli cells was greater at 38 C than at 34 C. It is concluded that Sertoli cells do not show in vitro any inhibitory effect of body temperature on any of the functions tested, but on the contrary, are more active at body than at scrotal temperature. The possible significance of the differences seen is discussed. PMID- 2985366 TI - Ovine pineal alpha 1-adrenoceptors: characterization and evidence for a functional role in the regulation of serum melatonin. AB - Plasma melatonin in sheep increases to nocturnal levels rapidly (10-20 min) after dark onset. This increase is blocked by iv prazosin (1 mg), but not propranolol (6 mg). Prazosin also blocks the elevation in pineal melatonin content after dark onset, but does not significantly alter the rise in N-acetyltransferase activity or the elevation in pineal N-acetylserotonin content. Since the nocturnal elevation in N-acetyltransferase, a neurally regulated event, was unaltered, this suggests that prazosin does not significantly impair the transmission of neural signals from the eye to the gland, but does act on pineal alpha 1-adrenoceptors to block melatonin production. This is supported by binding studies in ovine pineal membranes using [125I] iodo-2-[beta-(4 hydroxyphenyl)ethylaminomethyl]tetralone, which revealed that binding is rapid, reversible, saturable, and stereo-specific. Saturation studies indicated the presence of a single class of binding sites, with an equilibrium binding constant (Kd) of 32 +/- 6 pM and a maximum binding of 139 +/- 19 fmol/mg protein. The relative potencies of several adrenergic agonists and antagonists in competition studies indicated that the receptor belongs to the alpha 1-subclass of adrenoceptors. Together, these data suggest that melatonin synthesis in the sheep pineal gland is controlled in part by an alpha 1-adrenoceptor mechanism at a step beyond N-acetylation. PMID- 2985367 TI - In vitro 3,5,3'-triiodothyronine binding to rat cerebrocortical neuronal and glial nuclei suggests the presence of binding sites unavailable in vivo. AB - The maximal binding capacity (MBC) of the rat cerebrocortical nuclear T3 receptor, as determined by in vivo saturation techniques, is approximately half that measured in vitro on isolated nuclei or solubilized receptors. To investigate this disparity, the MBC values determined in vivo and in vitro for both rat cerebral cortex and liver were compared, taking into account nuclear receptor loss or inactivation and the presence of endogenous T3. By Scatchard analysis of T3 binding to isolated nuclei in vitro at 37 C, the uncorrected MBC values (mean +/- SEM; n = 3) for the cerebrocortical nuclear T3 receptor in euthyroid and hypothyroid rats were 0.80 +/- 0.14 and 0.66 +/- 0.07 ng T3/mg DNA, respectively, and were not significantly different. The Kd values were also not significantly different (5.6 +/- 0.3 and 5.2 +/- 0.9 X 10(-10) M, respectively). After corrections for incomplete dissociation and receptor inactivation under the in vitro conditions, the overall mean MBC increased by approximately 33% to 0.97 ng T3/mg DNA, or about 3.6 times the in vivo MBC. In addition, cerebrocortical nuclei prelabeled in vivo with +/- 131I]T3 at near-saturating levels and subsequently incubated with [125I]T3 in vitro at concentrations up to 10 times the Kd were shown to bind as much as 4 times more T3 in vitro relative to the amount of endogenous hormone which dissociated, thus exceeding the in vivo MBC by a factor of two. Parallel experiments with isolated liver nuclei did not show the existence of nuclear T3 receptors which were available only in vitro, even when the corrected MBC (0.77 ng T3/mg DNA) was compared with the MBC obtained by the in vivo saturation technique (0.76 ng T3/mg DNA). The experiments with liver nuclei were done at 25 C to reduce the rate of inactivation or loss of nuclear T3 receptors in this tissue. By fractionating isolated cerebrocortical nuclei into neuronal and glial subpopulations on discontinuous sucrose gradients, the high affinity, limited capacity nuclear T3 receptor could only be identified in the neuronal fraction. No consistent specific binding of T3 was observed in glial nuclei that were 80% pure, suggesting that either glial cells in the adult rat are not likely to be direct targets of thyroid hormone or that thyroid hormone may act via nonnuclear receptor-mediated pathways. We conclude that only neurons have specific high affinity, limited capacity nuclear T3 receptors and that as many as half of these receptors may not be accessible to plasma T3.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985368 TI - The effect of relaxin on cyclic adenosine 3',5'-monophosphate concentrations in rat myometrial cells in culture. AB - Relaxin, a uterine relaxant secreted by the corpus luteum, was able to elevate cAMP concentrations in the presence of 1-methyl-3-isobutyl xanthine (MIX) (0.1 mM) or forskolin (0.4 microM) in a time- and dose-dependent manner in rat myometrial cells in culture but not in stromal cells. The optimal culture conditions for the cAMP response were determined to be an initial plating density of 1-1.5 X 10(6) cells/ml (3 ml/35-mm dish) and a 2-day culture period. In the presence of MIX, the time course of cAMP elevation in response to relaxin exhibited a lag phase of more than 5 min before cAMP concentrations rose significantly. However, in the presence of forskolin, relaxin elevated cAMP within 1 min. The concentration-response relationships were almost identical in the presence of MIX or forskolin. Isoproterenol was able to increase cAMP concentrations in myometrial cultures in both the absence and presence of MIX and to elevate cAMP levels rapidly within 1 min. These data suggest that cAMP could play some role in the initiation of uterine relaxation mediated by relaxin. PMID- 2985369 TI - Stimulation of adenylate cyclase activity in rat thymocytes in vitro by 3,5,3' triiodothyronine. AB - In view of our previous demonstration that T3 promptly increases the cAMP concentration in freshly isolated rat thymocytes in vitro, we studied the effects of T3 on adenylate cyclase activity in a crude thymocyte plasma membrane preparation. In common with adenylate cyclase in other tissues, the enzyme in rat thymocytes was activated by NaF, GTP, 5'-guanylylimidodiphosphate, and beta adrenergic agonists and was inhibited by high concentrations of calcium. In the presence of 1 microM Ca+2, T3 induced a time-dependent increase in adenylate cyclase activity that was statistically significant between 1 and 2 min and maximum between 2 and 5 min after hormone addition. As judged from observations made at 5 min, the effect of T3 was dose dependent over the range 1 nM to 1 microM. The stimulatory effect of T3 was calcium dependent, since it was abolished by EGTA at a concentration (0.5 mM) that did not alter basal enzyme activity, and the effect of T3 in the presence of EGTA was restored by the addition of either 0.1 or 1 mM Ca+2. As judged from the lack of hydrolysis of added cAMP, phosphodiesterase activity in the assay mixture was nil in both the presence and absence of T3. Both epinephrine and the specific beta-adrenergic agonist isoproterenol, but not the alpha-agonist phenylephrine, increased adenylate cyclase activity, and their effects appeared to be additive to that of T3. The beta-adrenergic antagonist L-alprenolol, in doses that did not influence basal adenylate cyclase activity, produced a dose-related inhibition of the stimulatory effect of T3 and of the effects of epinephrine and isoproterenol as well. Neither D-alprenolol nor the alpha-antagonist phentolamine had any effect. Various thyronine analogs displayed a rank order of potency in stimulating adenylate cyclase activity very similar to their relative potencies in increasing cAMP concentration in the intact thymocyte. These findings reveal that T3 stimulates adenylate cyclase activity in rat thymocyte plasma membrane preparations. With respect to calcium dependence, inhibition by alprenolol, and response to thyronine analogs, this effect has properties similar to those of the increase in cellular cAMP concentration induced by T3 in the intact thymocyte. It can be concluded, therefore, that the effect of T3 to increase 2-deoxyglucose uptake by the rat thymocyte in vitro, a response consequent to an increase in thymocyte cAMP concentration, derives from a stimulatory effect of the hormone on adenylate cyclase itself. PMID- 2985370 TI - L-Triiodothyronine (T3) stimulates growth of cultured GC cells by action early in the G1 period: evidence for mediation by the nuclear T3 receptor. AB - Incubation with T3 results in a dose-dependent increase in growth rate of cultured GC cells, a GH-producing rat pituitary tumor cell line. The T3-induced increase in growth rate results mainly from shortening of the G1 period from 79.4 +/- 4.3 (SD) h in cells grown in T3-depleted medium (-T3) to 10.0 +/- 0.9 h. This effect can also be demonstrated in synchronized populations. Addition of T3 (0.3 nM) to cells synchronized in early G1 in the absence of T3 shortened the G1 period, estimated from graphic data, from more than 40-50 h to 13.4 +/- 2.1 h (n = 7). To determine the mechanism of this T3 effect, GC cells were grown in Dulbecco's modified Eagle's medium containing 10% serum plus or minus T3 (0.3 nM) and synchronized at the beginning of the G1 period by mitotic selection. Mitotic cells (85-100%), obtained by controlled mechanical shaking, were isolated by centrifugation and replated. The end of G1 was determined by the onset of DNA synthesis with [3H]thymidine as assessed by autoradiography (percent labeled nuclei). L-T3-induced shortening of G1 was detectable at 0.05 nM T3, half-maximal at physiological T3 (0.17 nM), and maximal between 0.3 nM and 1.0 nM T3. Addition of cycloheximide, 0.025 microgram/ml or 1.0 microgram/ml, decreased protein synthesis by 50% and 90%, respectively, and attenuated the T3 effect on G1 by 80 90%. The attenuation of the T3 effect on G1 by cycloheximide at a dose which inhibited protein synthesis suggests that T3-induced shortening of G1 may require new protein synthesis. Since glucocorticoids decrease the effect of T3 on induction of alpha-aminoisobutyric acid transport, their effect on T3-induced shortening of G1 was determined in G1-synchronized GC cells and in asynchronous cultures. Cortisol, 100 nM, significantly decreased the growth rate of asynchronous GC cells and attenuated the effect of T3 in G1-synchronized cells. Finally, T4 also decreased the length of G1 in a dose-dependent manner with a half-maximal effect at 40.0 nM. The half-maximal effect of T4 occurred at a nuclear iodothyronine concentration that was comparable to that achieved in incubations with 0.17 nM T3 (half-maximal dose). Thus, half-maximal shortening of G1 in synchronized GC cell cultures occurred at iodothyronine concentrations required for half-maximal occupancy of nuclear T3 receptors and for half-maximal induction of GH synthesis, growth rate, alpha-aminoisobutyric acid uptake, and depletion of the nuclear T3 receptor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985371 TI - Inhibition of hormone-induced steroidogenesis during cell proliferation in serum free cultures of rat granulosa cells. AB - Long term cultures of rat granulosa cells were grown in serum-free medium, consisting of Dulbecco's modified Eagle's medium mixed 1:1 with Ham's nutrient F 12 medium and supplemented with insulin, transferrin, hydrocortisone, and fibronectin (4F medium). In sparse cultures (10(4) cells/cm2), the granulosa cells were steroidogenically responsive to ovine FSH (NIADDK-oFSH-15) during days 1-2 and 10-14 (responsive periods). The major steroids produced were 20 alpha hydroxyprogesterone (20 alpha-OH-P) and 5 alpha-pregnane, 3 alpha,20 alpha-diol (pregnanediol). However, as of day 3, the cells gradually lost their steroidogenic responsiveness which was inhibited by 88% at day 7 (refractory period). Nevertheless, from day 8 onward, the cells regained their responsiveness which was fully restored at day 12. The transient loss of responsiveness was uniquely associated with progestin biosynthesis, since FSH-induced aromatase activity declined to background levels within 12 days and was never restored again. The loss of progestin responsiveness was not due to lack of cAMP because FSH induced increasing levels of cAMP accumulation, reaching maximal values on day 7 in culture. On the other hand, the onset of the refractory period occurred concomitantly with the entry of the cultured cells into a synchronous proliferation phase, during which the cell population doubled. Thereafter, as DNA synthesis ceased, the cells regained their steroidogenic responsiveness. A deliberate arrest of cell replication, in the presence of excess thymidine or in high density cultures, prevented the temporal loss of activity. The data presented favor the notion that cell proliferation and expression of differentiated functions are inversely related. It is suggested that growth related processes suppress steroidogenesis by an as yet unknown mechanism. PMID- 2985372 TI - The frequency of gonadotropin-releasing hormone stimulation determines the number of pituitary gonadotropin-releasing hormone receptors. AB - Gonadotropin-releasing hormone (GnRH) induces both synthesis and release of pituitary gonadotropins, but rapid or slow frequencies of stimulation result in reduced LH and FSH secretion. We determined the effects of frequency of GnRH stimulation on pituitary GnRH receptors (GnRH-R). Castrate male rats received testosterone implants (cast + T) to inhibit endogenous GnRH secretion. GnRH pulses were injected by a pump into a carotid cannula and animals received GnRH (25 ng/pulse) at various frequencies for 48 h. In control animals (saline pulses) GnRH-R was 307 +/- 21 fmol/mg protein (+/- SE) in cast + T and 598 +/- 28 in castrates. Maximum GnRH-R was produced by 30-min pulses and was similar to that seen in castrate controls. Faster or slower frequencies resulted in a smaller GnRH-R response and GnRH given every 240 min did not increase GnRH-R over saline controls. Equalization of the total GnRH dose/48 h (6.6 ng/pulse every 7.5 min or 200 ng/pulse every 240 min) did not increase receptors to the maximum concentrations seen after 30-min (25 ng) pulses. Serum LH responses after 48 h of injections were only present after 30-min pulses, and peak FSH values were also seen after this frequency. Serum LH was undetectable in most rats after other GnRH frequencies, even though GnRH-R was increased. These data show that GnRH pulse frequency is an important factor in the regulation of GnRH-R. A reduction of GnRH-R is part of the mechanism of down-regulation of LH secretion by fast or slow GnRH frequencies, but altered frequency also exerts effects on secretory mechanisms at a site distal to the GnRH receptor. PMID- 2985373 TI - Antilipolytic action of insulin: role of cAMP phosphodiesterase activation. AB - Exposure of 3T3-L1 adipocytes to 1 nM insulin for 10 min results in activation of particulate cAMP phosphodiesterase and suppression of lipolysis stimulated by 10 nM isoproterenol. When lipolysis was increased by cilostamide, a selective inhibitor of the particulate phosphodiesterase, the antilipolytic effect of insulin was not observed. Insulin did suppress lipolysis stimulated by Ro 20 1724, an inhibitor of soluble cAMP phosphodiesterase activity. Cilostamide did not interfere with insulin stimulation of glucose uptake, nor did it have any direct effect on cAMP-dependent protein kinase. Thus, inhibition of particulate but not soluble cAMP phosphodiesterase blocked the antilipolytic effect of insulin. Our findings support the idea that insulin inhibits lipolysis, perhaps in large part by activating particulate "low Km" cAMP phosphodiesterase, which seems to be functionally closely coupled with the hormone-sensitive lipase regulatory system influencing primarily a pool of cAMP utilized by the relevant protein kinase. PMID- 2985374 TI - TSH crosslinks to the TSH receptor through the beta subunit. AB - TSH binds specifically to the TSH receptor present on the surface of thyroid follicular cells and consequently activates adenylate cyclase. It was not known, however, which of the two subunits of the TSH bound to the receptor. By covalently crosslinking TSH to its receptor and characterizing the product with antisera specific for either the alpha or beta subunit of TSH, we have shown that the TSH beta subunit but not the TSH alpha subunit crosslinks to the TSH receptor. PMID- 2985376 TI - Diphenylbarbituric acid: its effects on neuromuscular and spinal cord function in the cat. AB - In studies of neuromuscular function in the cat soleus muscle, diphenylbarbituric acid (DPB; 10-80 mg/kg i.v.) depressed both the degree and the duration of posttetanic potentiation (PTP) in a dose-dependent manner. At doses of greater than 20 mg/kg, twitch strength was increased in both indirectly stimulated and directly stimulated chronically denervated preparations, indicating a direct effect of DPB on muscle. In the spinal cord, DPB (10-60 mg/kg) depressed both monosynaptic (2N) and polysynaptic discharges, with flexor and extensor reflexes being similarly affected. In addition, DPB was not selective for isolated 2N as compared with posttetanically potentiated monosynaptic responses. Thus, in its actions on neuromuscular function DPB resembles phenytoin; in the spinal cord the drug resembles phenobarbital. The data suggest that the capacity of a drug to curb high-frequency repetitive discharges is more important than curbing recruitment as exhibited by PTP of the 2N reflex. PMID- 2985375 TI - 1H and 15N NMR studies of protonation and hydrogen-bonding in the binding of trimethoprim to dihydrofolate reductase. AB - The binding of trimethoprim and [1,3,2-amino-15N3]-trimethoprim to Lactobacillus casei dihydrofolate reductase has been studied by 15N and 1H NMR spectroscopy. 15N NMR spectra of the bound drug were obtained by using polarisation transfer pulse sequences. The 15N chemical shifts and 1H-15N spin-coupling constants show unambiguously that the drug is protonated on N1 when bound to the enzyme. The N1 proton resonance in the complex has been assigned using the 15N-enriched molecule. The temperature-dependence of the linewidth of this resonance has been used to estimate the rate of exchange of this proton with the solvent: 160 +/- 10 S-1 at 313 K, with an activation energy of 75 (+/-9) kJ X mole-1. This is considerably faster than the dissociation rate of the drug from this complex, demonstrating that there are local fluctuations in the structure of the complex. PMID- 2985377 TI - Inducing potency of aryl hydrocarbon hydroxylase activity in human lymphoblastoid cells and mice by polychlorinated dibenzofuran congeners. AB - Aryl hydrocarbon hydroxylase (AHH)-inducing potency of eight polychlorinated dibenzofuran (PCDF) isomers, 3,4,5,3',4',5'-hexachlorobiphenyl (HCB) and 2,3,7,8 tetrachlorodibenzo-p-dioxon (TCDD) in two inbred mouse strains (AHH responsive and nonresponsive mouse strains) and eight human lymphoblastoid cell lines (four males and four females) was investigated to evaluate their relative toxic potency. In AHH nonresponsive DBA mouse strain, only TCDD induced hepatic AHH activity at a dose of 30 micrograms/kg, while in AHH responsive C57 mouse strain, six PCDF isomers besides TCDD could enhance the enzyme activity significantly. 2,3,7,8-Tetrachlorodibenzofuran (2,3,7,8-TCDF), 1,2,3,7,8-pentachlorodibenzofuran (1,2,3,7,8-PCDF) and 2,3,4,7,8-pentachlorodibenzofuran (2,3,4,7,8-PCDF) showed the highest AHH inducing activity among the PCDF isomers tested. In contrast with the results obtained from the mouse experiments, in human lymphoblastoid cells, 2,3,4,7,8-PCDF, 1,2,3,4,6,7-hexachlorodibenzofuran (1,2,3,4,6,7-HCDF) and 1,2,3,7,8-hexachlorodibenzofuran (1,2,3,4,7,8-HCDF) elicited the highest AHH induction and were as potent AHH inducers as TCDD. These observations suggest that toxicities of 2,3,4,7,8-PCDF, 1,2,3,4,6,7-HCDF and 1,2,3,4,7,8-HCDF in human tissues may be comparable to that of TCDD. It was also observed that in both male and female human cell lines, the degree of AHH inducibilities of these compounds were roughly parallel to that of 3-methylcholanthrene, possibly indicating that genetic susceptibility among human population to the toxic compounds are also present similar to those reported among mouse strains. PMID- 2985379 TI - Serum cyclic 3',5'-nucleotide phosphodiesterase in patients with various thyroid disorders. AB - Previous observations that cyclic 3',5'-nucleotide phosphodiesterase activity exists in mammalian sera including human serum prompted us to investigate the phosphodiesterase levels in sera of patients with various thyroid disorders. Both serum cyclic AMP phosphodiesterase (cAMP-PDE) and cyclic GMP phosphodiesterase (cGMP-PDE) activities measured in a low substrate concentration were elevated 3 fold in subacute thyroiditis and slightly in hyperthyroidism, compared to the normal. Slight decreases of these enzyme activities were observed in primary hypothyroidism. PDE activities were positively correlated with the value of T3 RSU and serum thyroid hormone levels in hyper- and hypothyroidism. Altered enzyme activities returned to normal during the course of recovery. Identical results were obtained when plasma was tested. These results suggest that serum PDE activities may be partly related to the thyroid function. PMID- 2985380 TI - Endoscopic and virological observations on respiratory disease in a group of young Thoroughbred horses in training. AB - A group of racehorses in training was examined on several occasions with a fibreoptic endoscope and monitored for viral infection. Only equine herpes virus 2 (EHV-2) infection was detected. Pharyngeal lymphoid hyperplasia (PLH) was present in all horses and decreased in severity with age. There was no association between PLH severity and antibody titres to EHV-1, or with the isolation of EHV-2. Finishing position in races was not affected by PLH severity. Exercise induced pulmonary haemorrhage (EIPH) was evident on 23 out of 49 (47 per cent) examinations after maximal speed training exercise. Eighteen out of 19 (95 per cent) horses examined on at least two occasions had EIPH but its occurrence was not predictable. Observable mucoid or mucopurulent exudate was present in the trachea in 60 out of 118 (50 per cent) examinations and the amount seen was increased following exercise. PMID- 2985381 TI - Plasma glucagon and catecholamines during exhaustive short-term exercise. AB - Plasma glucagon and catecholamine levels were measured in male athletes before and after exhaustive 15 min continuous running and strenuous intermittent short term exercise (3 X 300 m). Blood lactate levels were higher after the intermittent exercise (mean 16.7 mmol X 1(-1)) than after the continuous running (mean 7.1 mmol X 1(-1)). Plasma glucagon concentration increased during continuous running and intermittent exercise by 41% and 55%, respectively, and the increases in plasma noradrenaline concentration were 7.7- and 9.1-fold compared with the respective pre-exercise values. Immediately after the exercises plasma cyclic AMP, blood glucose and alanine levels were elevated significantly. The data suggest that the sympathoadrenal system is of major importance for liver glucose production during high-intensity exercises. Catecholamines directly stimulate liver glucose production and may indirectly stimulate it by enhancing the secretion of glucagon. PMID- 2985378 TI - Mechanism of action of toxic halogenated aromatics. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related halogenated aromatic hydrocarbons are a highly toxic class of environmental contaminants, as evidenced by numerous cases of accidental poisonings of human and animal populations and their extreme toxic potency in laboratory animals. The proposed model for the mechanism of action of TCDD and related compounds is analogous to that of the steroid hormones, which modulate gene expression through a receptor mechanism. In the steroid receptor model, the compound enters the cell cytoplasm where it acts as a specific ligand, binding selectively to a high affinity receptor protein. Bound to the appropriate ligand, the receptor concentrates in the nucleus where its increased association with chromatin leads to altered gene expression. This model has been useful in characterizing the Ah receptor; however, it does not provide a unifying hypothesis for all biochemical and toxic effects associated with exposure to halogenated aromatic hydrocarbons. Several findings suggest that a primary factor in determining TCDD toxicity might be tissue and species specific factors that control the actions of Ah receptor(s) in target tissues. Furthermore, numerous mechanisms might be involved. Clarifying the mechanism(s) for TCDD toxicity would enhance our ability to predict human health consequences to toxic halogenated aromatic hydrocarbons and would provide a more rational basis for risk analysis. PMID- 2985382 TI - Study on the mechanism of the appearance of noise effects. AB - This study was undertaken to investigate the dose-response relationship between the biological effect and noise exposure, and to consider the mechanism of the appearance of noise effects. Rats were exposed to noise at intensities of 60 dB (A), 80 dB (A) and 100 dB (A) for 240 min and examined for the change of activities of dopamine-beta-hydroxylase (DBH) in serum and adrenal glands. Plasma cyclic adenosine 3',5'-monophosphate (c-AMP) levels were also measured. Some rats were given 6-hydroxydopamine (6-OHDA) as a chemical sympathectomyzing agent 20 h before noise exposure in order to consider the mechanism of the appearance of noise effects. By noise exposure, serum DBH activity was significantly (P less than 0.01) increased at each intensity compared with the control group, but there were no remarkable changes in adrenal DBH activity. Plasma c-AMP level was also significantly elevated in response to the noise stress. When the rats, which had been pretreated with 6-OHDA, were exposed to noise with an intensity of 100 dB (A), the response of serum DBH activity was no longer observed. Therefore it is suggested that the effect due to noise exposure appears through the post ganglionic sympathetic nerve fiber. PMID- 2985383 TI - Cloning, nucleotide sequencing and expression of cDNAs encoding mouse urokinase type plasminogen activator. AB - Controlled extracellular proteolysis is catalyzed in part by the secretion of plasminogen activators. As a step in the study of the expression of these enzymes in mouse tissues, we have isolated five cDNAs encoding the mouse urokinase-type plasminogen activator from a cDNA library prepared with size-selected mRNA from MSV-transformed 3T3 cells. The longest cDNA insert contains the entire coding region of mouse urokinase, 58 base pairs of the 5' non-coding region, and the entire 3' non-coding region, which is 942 base pairs long. The deduced protein sequence, which starts with a signal peptide of 20 amino acids, shows extensive homology to that of human and porcine urokinase. However, in contrast to these enzymes, mouse urokinase contains no N-glycosylation site. Bacteria harbouring one of the recombinant plasmids synthesize and secrete into their periplasm a protease indistinguishable from mouse urokinase. PMID- 2985384 TI - The protein phosphatases involved in cellular regulation. 1. Modulation of protein phosphatases-1 and 2A by histone H1, protamine, polylysine and heparin. AB - The phosphorylase phosphatases in rat and rabbit liver cytosol that are markedly stimulated by histone H1, protamine and polylysine were identified as protein phosphatases-2A0, 2A1 and 2A2 by anion-exchange chromatography, gel-filtration and immunotitration experiments. Histone H1 and protamine also stimulated the dephosphorylation of phosphorylase kinase, glycogen synthase, fructose-1,6 bisphosphatase, pyruvate kinase, acetyl-CoA carboxylase and phenylalanine hydroxylase by phosphatases-2A1 and 2A2, and with several of these substrates activation was even more striking (20-100-fold) than that observed with phosphorylase (approximately 5-fold). Activation by basic polypeptides did not involve dissociation of these phosphatases to the free catalytic subunit. The dephosphorylation of phosphorylase by protein phosphatase-1 was suppressed by basic polypeptides, protamine and polylysine being the most potent inhibitors. However, the dephosphorylation of glycogen synthase, pyruvate kinase and acetyl CoA carboxylase were markedly stimulated by histone H1 and protamine (2-13-fold). Consequently, with the appropriate substrates, protein phosphatase-1 can also be regarded as a basic-polypeptide-activated protein phosphatase. Heparin stimulated (1.5-2-fold) the dephosphorylation of phosphorylase by phosphatases-2A0 and 2A1, provided that Mn2+ was present, but phosphatase-2A2 and the free catalytic subunit of phosphatase-2A were unaffected. Heparin, in conjunction with Mn2+, also stimulated (1.5-fold) the dephosphorylation of glycogen synthase (labelled in sites 3 abc), phosphorylase kinase and phenylalanine hydroxylase by phosphatase-2A1, but not by phosphatase-2A2. By contrast, the dephosphorylation of phosphorylase and phosphorylase kinase by protein phosphatase-1 was inhibited by heparin. However, dephosphorylation of glycogen synthase and pyruvate kinase by phosphatase-1 was stimulated by this mucopolysaccharide. The studies demonstrate that basic proteins can be used to distinguish protein phosphatase-1 from protein phosphatase-2A, but only if phosphorylase is employed as substrate. Optimal differentiation of the two phosphatases is observed at 30 micrograms/ml protamine or at heparin concentrations greater than 150 microM. PMID- 2985385 TI - The protein phosphatases involved in cellular regulation. 2. Purification, subunit structure and properties of protein phosphatases-2A0, 2A1, and 2A2 from rabbit skeletal muscle. AB - Protein phosphatases-2A0, 2A1 and 2A2 have been purified to homogeneity from rabbit skeletal muscle. Approximately 1 mg of phosphatase-2A0 and 2A1, and 0.5 mg of phosphatase-2A2, was isolated from 4000 g muscle within 10 days. Protein phosphatases-2A0 and 2A1 each comprised three subunits, termed A, B' and C (2A0) or A, B and C (2A1), while phosphatase-2A2 contained only two subunits, A and C. The A and C components of phosphatases-2A0, 2A1 and 2A2 had indistinguishable mobilities on sodium dodecyl sulphate/polyacrylamide gels and identical peptide maps. By these criteria, the C component was also identical to the catalytic subunit of phosphatase-2A purified from ethanol-treated muscle extracts. The electrophoretic mobilities of the B and B' subunits were slightly different, and their peptide maps were distinct. The molecular masses of the native enzymes determined by sedimentation equilibrium centrifugation were 181 +/- 6 kDa (2A0), 202 +/- 6 kDa (2A1) and 107 +/- 5 kDa (2A2), while those of the subunits estimated by sodium dodecyl sulphate/polyacrylamide gel electrophoresis were 60 kDa (A), 55 kDa (B), 54 kDa (B') and 36 kDa (C). These values, in conjunction with molar ratios estimated by densitometric analyses of the gels, suggest that the subunit structures of the enzymes are AB'C2 (2A0), ABC2 (2A1) and AC (2A2). Protein phosphatase-2A2 appears to be derived from 2A0 and/or 2A1 during purification through degradation or dissociation of the B' and/or B subunits. Protein phosphatases-2A0, 2A1 and 2A2 were the only phosphorylase phosphatases in rabbit skeletal muscle that were activated by the basic proteins, protamine (A0.5 = 0.25 microM), histone H1 (A0.5 = 0.3 microM) and polylysine (A0.5 = 0.04 microM). Activation by protamine varied over 5-20-fold for phosphatase-2A0 and 5 7-fold for phosphatases-2A1 and 2A2. The dephosphorylation of glycogen synthase was activated by basic proteins in a similar manner to the phosphorylase phosphatase activity. The isolated C subunit was also stimulated by histone H1 and protamine, but 5-10-fold higher concentrations were required, and with phosphorylase as substrate, maximum activation was only about 2-fold. Activation by basic proteins appears to involve their interaction with the A and/or C subunits, but not with the B or B' subunits, or substrates phosphorylase and glycogen synthase. PMID- 2985386 TI - Glycerol formation after the breaking of dormancy of Phycomyces blakesleeanus spores. Role of an interconvertible glycerol-3-phosphatase. AB - The breaking of dormancy of Phycomyces blakesleeanus spores by a heat shock was followed by a transient production of glycerol, which culminated within 5-10 min and was terminated at 20 min. Extracts of spores contained a magnesium-dependent glycerol-3-phosphatase active on both L-glycerol 3-phosphate and dihydroxyacetone phosphate but having more affinity for the first substrate than for the second. In extracts from dormant spores, the phosphatase was profoundly inhibited by physiological concentrations of inorganic phosphate, which induced cooperativity for the substrate, whereas the enzyme from heat-activated spores was much less inhibited and this difference in kinetic properties persisted after gel filtration of the enzymic preparation. When measured at 1 mM phosphate and 0.1 mM glycerol 3-phosphate, the phosphatase activity was undetectable in dormant spores, increased sharply during the heat treatment and the following 5 min at 25 degrees C, then fell again to a low value by 20 min. A similar transient activation of the enzyme was observed following the breaking of dormancy by incubation of the spores in the presence of 0.1 M ammonium acetate. Incubation of a cell-free extract or of the partially purified glycerol-3-phosphatase in the presence of ATP-Mg and the catalytic subunit of cyclic-AMP-dependent protein kinase released the enzyme from inhibition by phosphate and endowed it with the same kinetic properties as did the heat treatment of the spores. It appears therefore most likely that phosphorylation of glycerol-3-phosphatase by cyclic AMP-dependent protein kinase causes its activation and that this transient process explains the equally transient formation of glycerol by the spores after the heat shock. PMID- 2985387 TI - Radionuclide determination of right and left ventricular stroke volumes. AB - The relationship between radionuclide and thermodilution measurement of stroke volumes (SV) was investigated in 30 patients without valvular regurgitation or intracardiac shunt (group A) at rest and during exercise. Both attenuated radionuclide right ventricular (RV) and left ventricular (LV) SV measurements correlated well with the SV determined by the thermodilution method (r = 0.87 and r = 0.93, all P less than 0.001). The reliability of the radionuclide method to estimate SV was evaluated prospectively in two additional groups of patients. In 11 patients without valvular regurgitation or intracardiac shunt (group B) the radionuclide RVSV and LVSV closely approximated to thermodilution SV at rest and during exercise. In 15 patients with aortic regurgitation (group C) the radionuclide stroke volume ratio correlated well with the angiographic regurgitant fraction. Thus, both RVSV and LVSV and the severity of aortic regurgitation can be reliably measured with gated radionuclide ventriculography. PMID- 2985388 TI - Value of combined assessment of global and segmental ventricular contraction with right anterior oblique ECG-gated first-pass and left anterior oblique equilibrium radionuclide ventriculography. AB - A semi-automated, variable-region-of-interest method of analysis was used to measure both global and segmental left ventricular (LV) and global right ventricular (RV) contraction with ECG-gated first-pass and equilibrium radionuclide ventriculography. Normal values were defined in 20 healthy volunteers, and in 24 symptomatic patients, the results were compared with right anterior oblique (RAO) contrast left ventriculography. The global LV ejection fraction (LVEF) obtained by equilibrium imaging in the left anterior oblique (LAO) projection correlated closely with the results obtained by the gated first pass method in the RAO projection (r = 0.95) and those obtained with contrast left ventriculography (r = 0.94); furthermore, the interobserver variability was small (r = 0.985). The normal values for LVEF obtained using radionuclide techniques and contrast ventriculography did not differ, but with the equilibrium radionuclide method, the RV ejection fraction (RVEF) values were underestimated in comparison to those obtained by the RAO gated first-pass technique. In five patients with localised inferior segmental akinesis at contrast angiography, the RAO first-pass cine display demonstrated a corresponding wall-motion abnormality in all cases, but LAO equilibrium cine displays did so in only one out of five patients. For segmental quantitation of LV contraction, a computer programme defined the ventricular edge, divided the RAO LV images into five segments and determined both the segmental area contraction (SAC) and the counts-based segmental ejection fraction (SEF). Radionuclide SAC measurements correlated very strongly with SEF measurements (r = 0.94-0.99). Both radionuclide SAC and radionuclide SEF correlated well with contrast angiographic SAC, except in the inferobasal segment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985389 TI - Radionuclide assessment of right-ventricular involvement in inferior acute myocardial infarction: clinical correlations and in-hospital follow-up. AB - We studied 33 patients (28 males, 5 females) with first inferior acute myocardial infarction (AMI) and 12 normal volunteers. They underwent first-pass (FP) and equilibrium-gated radionuclide angiography (EGRA) within 4 days of the onset of symptoms. The parameters [ejection fraction (EF), peak ejection rate (PER), peak filling rate (PFR)] of the time-activity curve (TAC) of both ventricles [left ventricle (LV), right ventricle (RV)] were computed. The regional wall motion (RWM) was evaluated by parametric images (amplitude and phase). In 43% of the patients with AMI, we found a depressed RVEF, while the LVEF was decreased in only 33%. The sensitivity of diastolic parameters was shown to be clearly superior to that of the systolic parameters (RVPFR, 78%; LVPFR, 79%). The abnormalities of the overall performance of both ventricles were significantly correlated with those of the RWM. The abnormal RVEF allowed us to assess the necrotic involvement of the RV, while a depressed RVPFR suggested an impaired RV compliance mostly on an ischemic basis. Congestive heart failure (CHF) and shock syndrome were significantly correlated with depressed RV parameters, while the LVEF, despite RWM abnormalities, was in the normal range. EGRA with computation of the TAC parameters of both ventricles appeared to be useful for in-hospital prognostic evaluation, therapeutic planning and clinical follow-up of patients with inferior myocardial infarction. PMID- 2985391 TI - Cerebellar astrocytoma presenting as deterioration of handwriting in a child. AB - Deterioration of handwriting in an 11-year-old boy over a 2 month period was found to be caused by a cerebellar astrocytoma. The clinical picture was characterised by a lack of the classic symptoms of increased intracranial pressure. The only positive neurological findings pointed to an isolated right cerebellar symptomatology expressed by mild intention tremor and decreased tone of the right hand. Progressive deterioration of handwriting can be an ominous sign and it should be known to all professionals, as a lack of awareness can cause delay in expert referral and diagnosis. PMID- 2985390 TI - Concentration of 99mTc-Sn-N-pyridoxyl-5-methyltryptophan, a biliary agent, in distant metastases of hepatomas. AB - During the last 2 years, eight patients with hepatocellular carcinoma who were suspected of having distant metastases have been studied to determine whether a new biliary agent, 99mTc-Sn-N-pyridoxyl-5-methyltryptophan (99mTc-PMT), is taken up by extrahepatic tumors. In all eight patients, scintigrams showed a clearly increased uptake of 99mTc-PMT radioactivity by the extrahepatic tumors. In contrast, an increased uptake by the tumors of gallium citrate Ga 67 was only detected in four of the seven patients examined. The results obtained in this study suggest that 99mTc-PMT is useful both for characterizing the nature of extrahepatic tumors in patients with hepatoma and for detecting the metastases. PMID- 2985392 TI - Evaluation of four tests for detecting human rotavirus in feces. PMID- 2985393 TI - Biochemical study in 28 children with lactic acidosis, in relation to Leigh's encephalomyelopathy. AB - An enzymatic study of cultured skin fibroblasts was made in 28 patients with lactic acidosis. In three of these patients a diagnosis of Leigh's encephalomyelopathy was established from autopsy findings. Pyruvate decarboxylase (PDC) deficiency was found in four patients. In two of them, in whom Leigh's encephalomyelopathy was proved by autopsy, PDC activity was lower than 10% of the normal. The other two living patients, who showed 22%-25% of the normal activity, had clinical symptoms and courses different from Leigh's disease. These findings suggest that the patients with severe PDC deficiency develop Leigh's disease but those with mild deficiency may not. A deficiency of cytochrome c oxidase was found in two siblings. One of them, who was diagnosed as having Leigh's encephalomyelopathy by postmortem examination, showed a reduction of cytochrome c oxidase in the liver and brain. In the other sibling, who is living, the reduction of cytochrome c oxidase was demonstrated in the cultured skin fibroblasts and biopsied muscle. In an electron-microscopic study of biopsied muscle, two patients with mitochondrial myopathy were found. Their fundamental enzymatic defects were unclear. In two patients, in whom Leigh's disease was suspected following a brain CT, the production of 14CO2 from [3-14C] pyruvate was found to be low; suggesting a reduced activity of the TCA cycle. In another 18 patients, the fundamental defect was not clear. PMID- 2985394 TI - Isolated glucocorticoid deficiency: metabolic and endocrine studies in a 5-year old boy. AB - A 5-year-old boy is described who presented with episodes of hypoglycaemia triggered by mild infections or fever. Subnormal glucocorticoid production was confirmed by demonstrating low urinary excretion of free cortisol, low plasma cortisol concentrations that did not rise after glucagon and ACTH stimulation, and by elevated plasma ACTH levels. The selective nature of the abnormality was confirmed by demonstrating normal plasma electrolyte concentrations and blood pressure on a salt-restricted diet. Plasma renin activity and plasma aldosterone levels were also normal and responded appropriately to salt restriction and to frusemide-induced diuresis. Starvation-induced hypoglycaemia was associated with raised levels of blood ketone bodies and low blood alanine concentrations. Catecholamine secretion during hypoglycaemia was reduced. Glucocorticoid replacement therapy was effective in restoring normal glucose homeostasis. PMID- 2985395 TI - Influence of ACTH 4-10 and unilateral ECT on primary and secondary memory in depressive patients. AB - Primary and secondary memory were tested in depressive patients by means of immediate free recall of lists of words in a double blind intraindividual cross over comparison between ACTH 4-10 in a single s.c. dose and placebo, in connection with the second and third unilateral electroconvulsive treatment (ECT). The results of the study did not show any memory facilitating effect of ACTH 4-10 in immediate recall from primary and secondary memory, after ECT. The results indicate a negative effect of ECT on delayed recall from secondary memory 120 min after ECT. PMID- 2985396 TI - Solubility of inorganic kidney stone components in the presence of acid-base sensitive complexing agents. AB - Computation methods used to calculate the solubilities of the main inorganic kidney stone components in the absence or presence of acid-base-sensitive complexants as a function of pH are described. The solubilities of whewellite, brushite, whitlockite, octocalcium phosphate, hydroxyapatite and struvite were calculated in the presence of ethylenediaminetetraacetate (EDTA), trans cyclohexane-1,2-diaminetetraacetate (CDTA), citrate, tripolyphosphate and alpha hydroxyisobutyrate. The results explain why kidney irrigations with citrate give extremely variable results. The calculated solubility curves show that only EDTA and CDTA solutions with a pH of 8 or higher will have good litholytic properties for all compounds concerned. The computation methods presented in this paper can be used to calculate the solubility of a large number of salts. These salts may contain non-acid-base-sensitive cations capable of forming complexes, acid-base sensitive anions, hydroxyl ions, hydrogen ions or other acid-base-sensitive cations (e.g. NH4+), and the solubilities can be calculated in the presence or absence of acid-base-sensitive complexing agents. PMID- 2985397 TI - Superoxide anion generation by polymorphonuclear leukocytes enhanced in a patient with colony-stimulating activity-producing lung cancer. AB - We examined superoxide (O-2)-generating activity of polymorphonuclear leukocytes (PMN) from a patient with lung cancer in whom there was a marked granulocytosis. There was a high level of colony-stimulating activity (CSA) in both the serum and the supernatant of the culture of the primary tumor. The PMN from the patient produced almost three times more O-2 than did the PMN from healthy donors and other patients with lung cancers but with no granulocytosis. The binding of wheat germ agglutinin (WGA), used as a stimulus for O-2 generation, to the surface membrane of cells and the protein content in the cells were about double that noted in the case of healthy donors. Following total extirpation of the tumor, the level of CSA in the serum decreased, as did activity of PMN for O-2 generation, and the binding of WGA to the cells reverted to normal levels. PMID- 2985398 TI - Phenotyping of 60 cultured human gliomas and 34 other neuroectodermal tumors by means of monoclonal antibodies against glioma, melanoma and HLA-DR antigens. AB - The reactivity spectrum of three monoclonal antibodies (Mabs) to human malignant glioma, five Mabs to melanomas and one Mab anti-HLA-DR was investigated by an indirect antibody binding radioimmunoassay on a panel of cells derived from 60 glioma lines, including 47 malignant astrocytomas, 11 low-grade astrocytomas and two malignant ependymomas as well on cells from 12 melanoma, three neuroblastoma, three medulloblastoma, two schwannoma, two retinoblastoma, two choroid plexus papilloma, ten meningioma and 12 unrelated tumor lines. The anti-glioma Mabs BF7 and GE2 reacted preferentially with gliomas, while the anti-glioma Mab CG12 reacted with gliomas, melanomas, neuroblastomas and medulloblastomas. The five anti-melanoma Mabs reacted with gliomas, neuroblastomas and medulloblastomas. The anti-HLA-DR Mab D1-12 reacted with gliomas, melanomas and some meningiomas. On the basis of the data presented, we describe three different antigenic systems; the first one is glioma-associated, the second one is related to differentiation antigens expressed on cells derived from the neuroectoderm and the third is represented by HLA-DR antigens which are expressed not only on B-lymphoblastoid cells but also on melanomas and gliomas. PMID- 2985399 TI - Differences in basal and stimulated cyclic AMP content in calvaria bones from normal mice and mice with an impaired lysosomal function (beige mice). AB - Earlier we have shown that bone resorption is impaired in cultured calvaria from beige mice (an animal equivalent of the Chediak-Higashi syndrome in man). In the present study we have compared the concentrations of cyclic AMP and cyclic GMP in calvarial bones from beige mice with the nucleotide concentrations in bones from corresponding normal mice. In six independent experiments the basal concentrations of cyclic AMP was significantly (P less than 0.01) higher in bones from beige mice (on an average 50% augmented). The ratio of cyclic AMP/cyclic GMP was 2.43 times higher (P less than 0.01) in bones from beige mice. After stimulation with the phosphodiesterase inhibitor isobutylmethylxanthine and prostaglandin E2 no significant differences of cyclic AMP concentrations between beige and control mice could be registered. The response to adenosine was significantly higher (P less than 0.005) in bones from beige mice (4.3 +/- 0.4 fold of basal cyclic AMP concentrations, mean +/- SE) compared to control mice (1.9 +/- 0.4-fold of basal, mean +/- SE). The increased basal concentration of cyclic AMP in calvaria from beige mice may be due to increased sensitivity to some agonists, such as adenosine, rather than simply being a function of cell mass. PMID- 2985401 TI - Phorbol ester-induced expression and function of the interleukin 2 receptor in human B lymphocytes. AB - The expression of the interleukin 2 receptor (Tac) on normal B cells and Epstein Barr virus-transformed lymphoblastoid B cell lines is induced by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The receptor on B cells is indistinguishable from that on T cells by serological and biochemical analysis. It is functionally active so that TPA-induced B cells respond to interleukin 2 by increased DNA synthesis. These results suggest that the expression of Tac is not exclusively restricted to T cells and that interleukin 2 may play a role in B cell differentiation. PMID- 2985400 TI - Effect of renal function on renal responsiveness to parathyroid hormone in primary hyperparathyroidism and chronic renal failure. AB - The effect of renal function on the cyclic AMP (cAMP) response to exogenous parathyroid hormone (PTH) was examined in patients with chronic renal failure (n = 22) and primary hyperparathyroidism (n = 19). In the patients with chronic renal failure there was marked resistance to the effect of exogenous PTH. In primary hyperparathyroidism the cAMP responses were variable; most of the patients with an abnormally small response having impaired renal function. After parathyroidectomy, responsiveness improved to varying degrees. In three patients repeatedly tested up to several months after parathyroidectomy, the recovery of responsiveness was a gradual process which began within days but did not, however, return to normal. Thus, there was an irreversible component to the resistance to PTH in these patients. A strong negative correlation between plasma creatinine and the cAMP response to PTH (P less than 0.001) was found in a group of patients, some with treated primary hyperparathyroidism and some with chronic renal failure. Thus, renal impairment is an important, but probably not the sole, contributory factor involved in the irreversible resistance to the action of PTH in hyperparathyroidism. PMID- 2985402 TI - Specificity of P2-purinoceptor that mediates endothelium-dependent relaxation of the pig aorta. AB - ATP (EC50 5 microM) induced endothelium-dependent relaxation of the isolated aorta of the newborn pig, but the other naturally occurring nucleotides CTP, GTP, ITP and UTP were more than 100 times less potent. 2-Methylthio-ATP (EC50 0.1 microM) was 50 times more potent than ATP, but the unnatural enantiomers L-ATP and 2-methylthio-L-ATP were virtually inactive. beta,gamma-Imido-ATP and beta,gamma-methylene-ATP, both of which are resistant to degradation by ectonucleotidases on cultured pig endothelial cells, were much less potent than ATP. ADP beta S, which is also resistant to degradation, was equipotent with ATP at low concentrations but achieved a maximal relaxation of only 50% that of ATP. The Rp and Sp diastereoisomers of ATP beta S were both equipotent with ATP at low concentrations and both achieved approximately 60% of the maximal relaxation of ATP. The Rp and Sp diastereoisomers of ADP alpha S were both less potent than ATP and achieved only approximately 25% of the maximal relaxation of ATP. These results demonstrate that the P2-purinoceptor mediating endothelium-dependent relaxation of the pig aorta exhibits a high degree of specificity for the adenine base, is stereospecific for the D-ribofuranosyl moiety, requires a phosphate chain of 2 or 3 units but is not stereoselective toward this phosphate chain. These structural requirements have some features in common with the P2 purinoceptors on smooth muscle and on platelets, and are quite different from those of the ectonucleotidases present on pig endothelial cells in culture. PMID- 2985403 TI - Involvement of spot 35 protein, a cerebellar protein, in modulation of Purkinje cell activity of the rat cerebellum. AB - Spot 35 protein, applied iontophoretically, produced inhibition of spontaneous Purkinje cell activity in most cells and augmented the NE- or 5-HT-induced inhibition of spontaneous discharges whereas it hardly affected the inhibitory response of Purkinje cells to GABA, taurine, and beta-alanine, and the excitatory response to glutamate. It is suggested that spot 35 protein may be involved in normal physiological operation of the cerebellum through modulation of the NE or 5-HT system. PMID- 2985404 TI - Ca2+ antagonist receptor sites on human red blood cell membranes. PMID- 2985405 TI - Suppression by progabide of ethanol withdrawal syndrome in rats. AB - Progabide, a specific and clinically used GABA receptor agonist, was tested for its ability to suppress ethanol withdrawal syndrome. Male rats were rendered physically dependent on ethanol by feeding for 12 days on a liquid diet in which ethanol isocalorically replaced dextrose. Progabide (100-400 mg/kg i.p.), administered 8 h after ethanol was withdrawn, produced a dose-related inhibition of both tremors and audiogenically induced seizures. A single dose of 400 mg/kg of progabide completely suppressed all ethanol withdrawal reactions. Seizures were more sensitive to the drug than tremors. The results support the view that a decrease in GABA transmission plays a role in ethanol withdrawal symptoms and suggest that progabide may be tested as a possible treatment of ethanol withdrawal syndrome in man. PMID- 2985406 TI - Adrenergic hyposensitivity during long-term diuretic therapy--a possible explanation for the antihypertensive effect of diuretics? AB - The long-term effect of hydrochlorothiazide on beta 2-adrenoceptor density on mononuclear cells was investigated in 10 male patients with essential hypertension. There was a 40% reduction in beta 2-adrenoceptor density but no change in receptor affinity. This down-regulation of beta 2-adrenoceptors may explain the observed adrenergic hyposensitivity after long-term diuretic therapy. If lymphocytic beta 2-adrenoceptors represent presynaptic beta-adrenoceptors, a down-regulation of presynaptic beta 2-receptors may occur too, and result in a decrease of adrenergic transmitter release. Under this assumption long-term diuretic treatment causes its antihypertensive effect by modulating adrenergic sensitivity at the receptor level on the presynaptic side leading to an attenuated response to pressor hormones on the postsynaptic vascular side. PMID- 2985407 TI - Neuronal localization of substance P receptors in rat neostriatum. PMID- 2985408 TI - Benzodiazepine receptor photoaffinity labeling: correlation of function with binding. AB - Exhaustive photoaffinity coupling of flunitrazepam to living spinal cord neurons reduced the capacity of benzodiazepines to potentiate the electrophysiologically measured GABA response. In qualitative agreement with reversible binding data the dose-response curve for enhancement of the GABA response by benzodiazepines was shifted to the right, indicating that the remaining reversible benzodiazepine binding sites have lower affinity for benzodiazepines. Photoaffinity labeling did not reduce inhibition of the GABA response by beta-carbolines and there was only a small decrease in beta-carboline binding. In both control and photoaffinity labeled cultures, the inhibitory effect of beta-carbolines on the GABA response was reversed in the presence of excess benzodiazepine. The results indicate that the effects of photoaffinity labeling are confined to the BZD recognition site, and that coupling between benzodiazepine receptors and GABA receptors remains intact. PMID- 2985409 TI - Effects of Na+,K+-pump inhibitors and membrane depolarizing agents on acetylcholine-induced endothelium-dependent relaxation and cyclic GMP accumulation in rat aorta. AB - The purpose of this study was to investigate the effects of inhibitors of the Na+,K+-pump and membrane depolarizing agents on endothelium-dependent relaxation and elevated cyclic GMP levels induced by acetylcholine in rat thoracic aorta. Ouabain or exposure to K+-free or Mg2+-free Krebs-Ringer bicarbonate solution, agents and procedures known to inhibit the Na+,K+-pump, inhibited acetylcholine induced relaxation and the associated increased levels of cyclic GMP. However, the inhibitory effect of ouabain on cyclic GMP levels was abolished in the absence of norepinephrine or in the presence of norepinephrine and the alpha adrenergic receptor antagonist phentolamine. The membrane depolarizing agents KCl and tetraethylammonium also inhibited the acetylcholine-induced relaxation and the elevated cyclic GMP levels. Exposure to norepinephrine reduced the increased levels of cyclic GMP due to acetylcholine as compared to rested controls. This effect was inhibited by prior exposure to phentolamine, but not by the beta adrenergic receptor antagonist, propranolol. These results suggest that increased activity of the Na+,K+-pump may mediate, in part, endothelium-dependent relaxation; inhibition of relaxation may be due to membrane depolarization; the endothelium-dependent increased levels of cyclic GMP may increase Na+,K+-pump activity; a complex interaction exists between membrane polarization, the Na+,K+ pump and alpha-adrenergic stimulation in regulation of cyclic GMP accumulation and relaxation. PMID- 2985411 TI - Kinetics and displacement of [11C]RO 15-1788, a benzodiazepine antagonist, studied in human brain in vivo by positron tomography. AB - The brain regional distribution and kinetics of RO 15-1788, a benzodiazepine (BZD) antagonist labeled with 11C was studied by time-of-flight positron tomography after intravenous injection in four normal human volunteers. In two control studies, there was a high uptake of [11C]RO 15-1788 in gray matter structures initially (brain/blood ratio approximately 3), and subsequent retention that was highest in cerebral cortex, a structure known to have a high density of BZD receptors in vitro. Variation in tissue kinetics of [11C]RO among different gray matter structures may, however, suggest regional differences in binding characteristics or environment of BZD receptors. In two displacement studies, unlabeled RO 15-1788 was injected ten minutes after the radioligand: there was an immediate and marked washout of [11C]brain radioactivity that reached 70% in the occipital cortex with a 0.05 mg/kg dose (indicating a high specific to non-specific binding ratio) but was less prominent with a 0.01 mg/kg dose. These data suggest that [11C]RO 15-1788 may be useful for in vivo mapping of human brain BZD receptors using positron tomography. PMID- 2985410 TI - The aetiology of gastric ulceration induced by electrical vagal stimulation in rats. AB - Histamine and serotonin levels in gastric secretion and the effects of pharmacological antagonists were studied in rats in which stomach ulceration was induced by electrical vagal stimulation. Electrical vagal stimulation (2 and 5 V) produced a graded increase in haemorrhagic glandular mucosal ulcers. NaHCO3 perfusion completely neutralised the increased acid output but failed to prevent ulceration. Atropine inhibited gastric mast cell degranulation as well as histamine and serotonin release. Diphenhydramine, atropine and sub-diaphragmatic vagotomy antagonised the increase in intragastric pressure. Diphenhydramine, cimetidine, atropine or vagotomy but not methysergide reduced ulcer severity. It is concluded that gastric acid and serotonin do not play an important role in glandular ulceration induced by electrical vagal stimulation. The lesions probably result from increased intragastric pressure and release of gastric histamine which stimulates H1 and H2 receptors in the stomach. The similarities between the aetiologies of glandular ulcers due to electrical vagal stimulation and to stress are also discussed. PMID- 2985412 TI - Polymyxin B is a potent inhibitor of insulin hypoglycemia in mice. PMID- 2985413 TI - A cell line from pleomorphic adenoma of the rat salivary gland. AB - Pleomorphic adenoma of rat salivary gland is extremely rare, and culture of cells obtained from rat salivary gland tumors has not been reported. We have established a long-term cell line from a pleomorphic adenoma of a Sprague-Dawley rat submaxillary gland. The pleomorphic adenoma was composed of oval or spindle shaped cells occasionally forming a small duct. Alcian blue-positive intercellular matrices, consisting mainly of glycosaminoglycans, were abundant. The cultured cells showed characteristics similar to those of the original tumor. This cell line should be useful for biological and biochemical studies of glycosaminoglycan-synthesis of pleomorphic adenoma cells. PMID- 2985414 TI - Differences in molecular heterogeneity of glucagon-like immunoreactivity in plasma and liver metastases of a patient with alpha-cell tumor. AB - In the present study we characterized and compared the different molecular forms of glucagon-like immunoreactivity in extracts of peripheral plasma and hepatic metastases of a patient with pancreatic alpha-cell tumor. Plasma and tissue extracts were chromatographed on Sephadex G-50 columns. Immunoreactivity in the eluting fractions was assayed with an anti-glucagon antiserum that specifically recognizes the C-terminal region of the pancreas glucagon molecule. Total plasma glucagon-like immunoreactivity prior to surgery was 26.64 nmol/l and consisted of four peaks of immunoreactivity of apparent 9,000 mol wt, 5,800-5,400 mol wt, and 4,000 mol wt. Total glucagon-like immunoreactivity extracted from the hepatic metastasis was 47.41 nmol/g wet weight and eluted as two major peaks of immunoreactivity as follows: peak I, mol wt 3,800, corresponding to "true" 3,500 mol wt glucagon; peak II, mol wt 1,400, probably consisted of glucagon degradation products. The results clearly demonstrated that both plasma and glucagon-like immunoreactivity extracted from hepatic metastases were heterogeneous and comprised species corresponding not only to "true" glucagon but also to higher mol wt forms. The lack of higher mol wt forms of immunoreactivity in the hepatic metastases of the alpha-cell tumor suggests that this metastatic tumor tissue may contain an enzyme capable of converting the higher mol wt forms to smaller glucagon-like components whereas this degradative system seems to be defective in the primary tumor. PMID- 2985415 TI - Stem cell defects after cytoreductive therapy in man. AB - The study of stem cell defects in man after cytoreductive therapy appears to be of particular importance since hematotoxicity represents the major limiting side effect in many instances. On the other hand, such studies are met by difficult methodological problems. In particular, bone marrow sampling in man cannot be done on a quantitative basis. Furthermore, ethical considerations restrict the sampling of serial bone marrow specimens. We have studied the reaction of hematopoiesis including stem cells in man with the available methods, in particular CFU-GM (CFU-C), CFU-E, and BFU-E in bone marrow (BM) and peripheral blood (PB) during the course of various cytostatic regimens given for adjuvant and palliative chemotherapy of human cancer. Major findings of these studies were The acute reaction of BM stem cells and differentiated BM precursor pools corresponds to the mechanisms known to be effective in animal experiments. The PB CFU-GM may be of particular importance in man in demonstrating long-term derangements. In an attempt at quantification of human BM data, an index was developed from BM spicule morphometry and from standardized BM aspirate cellularity. Stem cell defects in BM and PB were observed that did not show up in the PB counts, in particular after nitrosoureas. Indication of prolonged derangements in stem cells up to five years after chemotherapy were observed after adjuvant therapy for breast cancer. From these data, it appears advisable to study stem cell data in man for newly developed chemotherapeutic regimens in order to minimize early and late side effects on hematopoiesis. PMID- 2985416 TI - Relationship between myelosuppression and chemotherapeutic response in small cell bronchogenic carcinoma. AB - Most cancerocidal agents have myelosuppression as their major toxicity. In some clinical studies it has been possible to show a relationship between the amount of administered drug and the therapeutic efficacy. Within any defined protocol, however, there may be much variability in the severity of myelosuppression. We attempted to determine whether the tumor response might be related to this toxicity. We evaluated a total of 177 patients with small cell bronchogenic carcinoma, treated by five successive regimens of combination chemotherapy, consisting of either cyclophosphamide and vincristine alone or with doxorubicin or doxorubicin plus bacillus Calmette-Guerin (BCG) or doxorubicin plus methotrexate, for a number of prognostic factors (age, sex, extent of disease, performance status, sites and number of metastases, serum LDH and alkaline phosphatase, weight loss, leukopenia, and thrombopenia). Leukopenia (mean 415 +/- 478/mm3, range 0-2000/mm3) had a weak influence on the incidence of complete remission, which was highest with the least severe nadir (P = 0.027). Thrombopenia was a nonsignificant factor (P = 0.738). Both leukopenia and thrombocytopenia had no influence on the overall survival. Because these drug combinations were based on cyclophosphamide, which requires metabolic activation, we evaluated the relationship of myelosuppression and the incidence of response in a second group of patients with small cell bronchogenic carcinoma treated with a VP16, cyclophosphamide, doxorubicin, vincristine sulfate protocol. In this analysis, no relationship could be detected between remission and myelosuppression. Granulocytopenia or thrombocytopenia also-showed no significant influence on the achievement of long-term survival beyond 36 months. PMID- 2985417 TI - Patterns of cytochrome oxidase activity in areas 17, 18 and 19 of the visual cortex of cats and kittens. AB - The laminar pattern of cytochrome oxidase activity was studied in visual cortical areas 17, 18 and 19 in adult cats and kittens, following electrophysiological determination of the boundaries of these areas in all but the youngest animals. The patterns of cytochrome oxidase staining and the cytoarchitectonic appearances of areas 17, 18 and 19 were compared. At all ages activity was especially high in the region of layers IV and VI in areas 17 and 18, and was low in all laminae in area 19. The results suggest that the degree of cytochrome oxidase activity in these regions of the visual cortex may be related to the strength and type of projection that they receive from the lateral geniculate nucleus. The cytochrome oxidase technique is a useful means of defining the 18/19 border, and may help locate the boundary between areas 17 and 18, in both adult cats and kittens. PMID- 2985418 TI - Interaction between inhibitory pathways to principal cells in the lateral geniculate nucleus of the cat. AB - Inhibitory interactions between interneurones of the lateral geniculate nucleus (LGN) of the cat were studied with an indirect method based on intracellular recordings of synaptic responses in principal cells. Recurrent inhibitory postsynaptic potentials (IPSPs), evoked by antidromic activation of principal cell axons in the visual cortex, were depressed by a preceding stimulation of the optic tract or the visual cortex. Disynaptic feed-forward IPSPs, evoked by optic tract stimulation, were likewise depressed after cortex stimulation. The duration of the depression was in both cases about 100 ms. The effect was not due to conductance changes in the recorded principal cells or to activation of cortico geniculate fibres. The observations indicate that perigeniculate neurones, the recurrent inhibitory interneurones of the LGN, have mutual inhibitory connexions and that they also project to intrageniculate interneurones, the inhibitory cells in the feed-forward pathway to principal cells. These conclusions were supported by intracellular recordings from a few interneurones. No evidence was found for interaction between feed-forward interneurones activated from separate eyes or for a projection from intrageniculate interneurones to perigeniculate cells. The results point to an unexpected similarity in the organization of the recurrent inhibitory system of principal cells in the LGN and of spinal motoneurones. It is suggested that the recurrent system of the LGN serves as a variable gain regulator in analogy with a recently proposed model for the spinal system. PMID- 2985419 TI - Nociceptive neurones in the superficial dorsal horn of cat lumbar spinal cord and their primary afferent inputs. AB - The morphology, background activity and responses to stimulation of primary afferent inputs of small neurones in the superficial dorsal horn which could only be excited from the skin by noxious stimulation were investigated by intracellular recording and ionophoresis of HRP. Neurons which gave similar responses to afferent stimulation were morphologically heterogeneous with respect to dendritic tree geometry and axonal projection, but were located around the lamina I/II border. Cutaneous excitatory receptive fields responding to noxious stimulation were generally small; most neurones had more extensive inhibitory fields responding to innocuous mechanical stimulation, in many cases overlapping the excitatory fields. Generally, stimulation of the excitatory field resulted in depolarization of the neurone and increased action potential firing, and stimulation of the inhibitory field resulted in hyperpolarization. Electrical stimulation of peripheral nerves revealed the existence of converging excitatory inputs carried by different fibre groups, and all neurones received an inhibitory input activated at low threshold. Excitatory responses were short-lived and occurred consistently in response to repeated stimulation. Central delay measurements gave evidence of a number of A delta monosynaptic inputs but only one A beta monosynaptic input; inhibitory inputs along A beta fibres were polysynaptic. The constant latency and regularity of the C response suggested monosynaptic connections. Low intensity stimulation of inhibitory inputs elicited a short-lived i.p.s.p. which increased in amplitude with increasing stimulus strength until it disappeared into a more prolonged hyperpolarization. This was associated with inhibition of background action potentials, and increased in duration with increasing stimulus strength up to C levels, indicating an A delta and C component. It is suggested that the level of excitability of these neurones depends on the relative amounts of concurrent noxious and innocuous stimulation, and that the resultant output, which is conveyed mainly to other neurones within the spinal cord, could modulate reflex action at the spinal level as well as affecting components of ascending sensory pathways. PMID- 2985420 TI - Horseradish peroxidase cross-reacts during cytochrome oxidase histochemistry. AB - Injections of horseradish peroxidase conjugated with wheat germ agglutinin (WGA HRP) were made into the cortex of kittens. When cytochrome oxidase histochemistry was performed on sections taken through the injection sites staining of the WGA HRP deposits occurred, demonstrating that WGA-HRP can cross-react during processing for cytochrome oxidase. PMID- 2985421 TI - The dynamic sensitivity of pyramidal tract neurones is influenced by previous activity. AB - The dynamic sensitivity of Pyramidal Tract (PT) neurones has been examined in silent cells and after pre-activation with an action potential evoked at different intervals before the discharge onset. In the resting conditions the dynamic sensitivity increases after the first interspike interval of the repetitive discharge. When a conditioning spike precedes the discharge onset by 50 ms (i.e. of a time sufficient for its afterhyperpolarization, AHP, to fade out) the dynamic sensitivity becomes uniform throughout the ramp. This linearizing effect progressively declines when the conditioning interval is increased to 100 and 150 ms and is attributed to the "depression" induced by the conditioning spike on the AHP of the first spike of the ramp discharge. In natural conditions, the effect would be entirely developed when the neurones fire at their minimal discharge rate (1/AHP duration). PMID- 2985422 TI - Dysfunction of the midbrain angular complex can accentuate or attenuate circling behaviour in the rat. AB - The role of the midbrain angular complex (AC) in the execution of motor behaviours was investigated in the rat. In an automated holeboard apparatus bilateral AC electrolesions attenuated exploration and increased locomotor performance of drug-free rats on the first and second test occasions respectively; the latter result may signify a retarding of between-session habituation. Apomorphine also decreased locomotion and almost abolished head dipping and rearing in the holeboard; bilateral AC lesions reinstated locomotion to a normal level without modifying the other behavioural parameters. An electrolesion of one AC did not affect the animal's posture or spontaneous locomotion in the open field, but gave rise to pronounced ipsiversive circling when coupled with systemic administration of apomorphine. In unilaterally 6 hydroxydopamine (6-OHDA) treated rats subcutaneous injection of apomorphine evoked robust contraversive circling. A concomitant lesion of the ipsilateral AC introduced an additional ipsilateral bias to these animals' movements; contraversive circling was initially curtailed and posture reduced (or reversed), while stereotyped activities (particularly grooming) were suppressed. Contralateral orientation and circling were restored by subsequently lesioning the contralateral AC as well; bilateral AC lesions significantly potentiated circling to systemic apomorphine. Contralateral locomotor asymmetry was also produced by depositing apomorphine stereotaxically into the supersensitive caudate, or by microinjecting one substantia nigra zona reticulata with muscimol (in naive rats). Both rotational responses were facilitated by injury to the ipsilateral AC. The effects of electrocoagulating the AC were generally duplicated by discrete microinjection of muscimol or gamma-vinyl GABA into this area, suggesting GABA-mediated synapses are normally operative in this part of the brain. These results do not support the claim that the AC is specifically engaged in mediating postural asymmetry in the unilaterally 6-OHDA denervated rat. Instead, we believe that impairment of neurotransmission through one AC imposes an independent and reciprocal tendency to move towards that side of the brain, as well as attenuating stereotypy and facilitating locomotion. The resultant behavioural response to systemic apomorphine shown by animals bearing these two types of lesion embodies these separate actions. PMID- 2985423 TI - Histochemical assessment of cytochrome oxidase activity for monitoring ischemic muscle injury. AB - Electromyographic detection of increased neuromuscular conduction latency that follows electrostimulation of spinal nerves has been used widely as a clinical tool for detection of ischemic muscle injury. We hypothesized that a biochemical marker for stagnant hypoxia provides more direct evidence for evaluating severity and monitoring resolution of ischemic muscle injury than the EMG. To detect intracellular changes in oxygen use during experimental ischemia, ultrathin sections of rat gastrocnemius muscle were treated with reducing agent, 3,3' diaminobenzidine (DAB), a histochemical marker for intramitochondrial cytochrome oxidase activity. The observed decrease in mitochondrial uptake of DAB suggested that a decrease in cytochrome oxidase activity was associated with experimental ischemia. Neuromuscular conduction latency in rat gastrocnemius muscle was also quantitated after electrostimulation of the sciatic nerve. Ligation of the femoral artery produced ischemic tissue injury, during which recordings of the EMG showed that conduction latency increased [from a mean +/- SEM control value of 3.09 +/- 0.13 to 3.92 +/- 0.22 ms (N = 11, P less than 0.001).] The changes in both histochemically detectable cytochrome oxidase activity and neuromuscular conduction latency were reversed by reperfusion. Response of the rat tissue to arterial occlusion was thereby shown to be a physiologic model for skeletal muscle response to ischemia. In addition, histochemical detection of cytochrome oxidase activity was shown to be a sensitive intracellular marker for decreased oxygen use during ischemic muscle injury. PMID- 2985425 TI - Blood pressure lowering effect of eicosapentaenoic acid-rich diet in normotensive, hypertensive and hyperlipemic subjects. AB - A mackerel diet or a herring diet in which two cans of fish fillet were consumed daily over 2 weeks within a prescribed regimen, in a crossover design, were given to 15 normotensive volunteers, 14 patients with mild essential hypertension and eight patients with type IV and V hyperlipoproteinemia (HLP). In normotensives a markedly lower systolic and diastolic blood pressure at the end of the period on the mackerel diet could be observed, whereas in hypertensive and hyperlipemic subjects only systolic blood pressure was significantly decreased. After the herring diet, which served as control, changes in blood pressure were of a minor degree. PMID- 2985426 TI - Involvement of reactive oxygen species in the microsomal S-oxidation of thiobenzamide. AB - Superoxide dismutase, catalase and methional proved capable of inhibiting the microsomal oxidation of thiobenzamide, which is most probably catalyzed by the flavin-containing monooxygenase. This indicates that excited oxygen species (e.g. X O-2,H2O2, X OH) are involved in the catalytic cycle of this enzymatic reaction. CO, which inhibits the cytochrome P-450-dependent oxygen radical formation, had no effect on the oxidation reaction, suggesting that the source of the reactive oxygen species is not the microsomal mixed-function oxidase. PMID- 2985427 TI - Interaction of BW755C, a potent inhibitor of lipoxygenase and cyclo-oxygenase, with mitochondrial cytochrome oxidase. AB - The interaction between BW755C (3-amino-1-[m-(trifluoromethyl)phenyl]-2 pyrazoline), a potent inhibitor of both lipoxygenase and cyclo-oxygenase, and respiratory chain in mitochondria and electron transport particles (ETP) from rat livers was examined. BW755C accelerated the oxygen uptake by mitochondria without the addition of substrate for the respiratory chain. Spectrophotometric study revealed that BW755C was quickly oxidized by cytochrome oxidase in mitochondria to a compound possessing an absorption maximum at 524 nm. p-Phenylenediamine (p diaminobenzene, PPDA), which, like BW755C, serves as an electron donor to cytochrome oxidase, was shown to inhibit the generation of active oxygen in macrophages; the inhibition was stronger than that of BW755C. These results strongly suggest that the oxidative conversion of BW755C by mitochondrial cytochrome oxidase is associated with its potentially inhibitory action on the active oxygen-generating system in phagocytes. PMID- 2985428 TI - Selective oxidative destruction of iron-sulfur clusters. Ferricyanide oxidation of Azotobacter vinelandii ferredoxin I. AB - The destructive oxidation of aerobically isolated 7Fe Azotobacter vinelandii ferredoxin I [(7Fe)FdI] by Fe(CN)3-6 is examined using low-temperature magnetic circular dichroism (MCD) and EPR. The results demonstrate that oxidation of the [3Fe-3S] cluster occurs only after essentially complete destruction of the [4Fe 4S] cluster. It is therefore feasible by controlled Fe(CN)3-6 oxidation to obtain a partially metallated form of FdI, (3Fe)FdI, containing only a [3Fe-3S] cluster. The MCD and EPR data demonstrate that the [3Fe-3S] cluster in (3Fe)FdI is essentially identical in structure to that in the native protein. PMID- 2985424 TI - Etiology of rheumatoid arthritis. AB - Definite genetic associations with immunological cooperative HLA-D(R) antigens have been demonstrated for rheumatoid arthritis (RA). Microbial etiology has not been proven, but some hope for the supporters of this view is still given by small viruses, plasmids of enteric bacteria or perhaps oncogen-like DNA sequences. Yet, electrophoretical analysis of membrane proteins or surface glycoproteins of RA synovial cells does not show any differences compared to reference cells. Autoimmunity to several tissue elements has been demonstrated, but most of it is of secondary nature. Antigenicities of type II and III collagens are probably only contributory factors for HLA-DR4 positive individuals. Proteoglycans or minor cartilage collagens have not been extensively studied, so far. Endocrine, dietary or psychological influences might be triggering events for otherwise 'preloaded' individuals. PMID- 2985429 TI - Molecular mechanisms regulating the synthesis of transferrin receptors and ferritin in human erythroleukemic cell lines. AB - The effects of either iron salts or iron chelators on the biosynthesis of transferrin receptors in human erythroleukemic lines was investigated. Addition of these compounds induced a rapid and marked decrease (iron salts) or increase (iron chelators) in the level of transferrin receptors synthesis. Both phenomena were inhibited by actinomycin D. In contrast, the increase in the synthesis of ferritin induced in these cells by addition of iron salts was not inhibited by actinomycin D. These results suggest that iron salts modulate the synthesis of transferrin receptors and ferritin via different molecular mechanisms, of transcriptional and translation type, respectively. PMID- 2985430 TI - Potentiation of the antagonistic effect of ACTH11-24 on steroidogenesis by synthesis of covalent dimeric conjugates. AB - Covalent dimerization of the adrenocorticotropin fragment ACTH11-24 increases its antagonistic potency on the ACTH-induced steroidogenesis in isolated bovine fasciculata/reticularis cells by 3 orders of magnitude when the C-termini are linked via a 10 A long spacer. This strong potentiation, probably mediated by cross-linking of the receptors, was shown to be dependent on the point of attachment of the monomeric fragment to the spacer, thus providing information about the position of the binding site in the hormonal segment and about the distance of the receptors on the cell surface. PMID- 2985431 TI - The oxidation of exogenous cytochrome c by mitochondria. Resolution of a long standing controversy. AB - Several reports in the past have dealt with the oxidation of cytochrome c added to suspensions of rat liver mitochondria. Yet, it is generally believed that the cytochrome cannot penetrate the outer membrane. Probably it has been assumed that the permeability of the outer membrane to cytochrome c is very low but finite, and that fast oxidation may be observed if time is allowed for sufficient penetration before initiation of electron flow. Here we show that this view is false. The main fraction of rat liver mitochondria, as isolated by conventional procedures, does not catalyse any significant oxidation of added cytochrome c, even after prolonged incubation. The observed appreciable oxidation of added cytochrome c is catalysed by a very small fraction (5-12%) of the mitochondria that apparently has a damaged outer membrane. Consequently, the turnover of cytochrome oxidase is very high in this fraction during oxidation of added cytochrome c. This finding readily explains why Moyle and Mitchell (e.g., FEBS Lett. 88 (1978) 268-272; 90 (1978) 361-365) have failed to observe proton translocation by cytochrome oxidase during oxidation of ferrocytochrome c added to rat liver mitochondria, which has been their main reason for rejecting the proton-pumping function of cytochrome oxidase. PMID- 2985432 TI - Pertussis toxin inhibits chemotactic factor-induced phospholipase C stimulation and lysosomal enzyme secretion in rabbit neutrophils. AB - Pertussis toxin suppressed [32P]polyphosphoinositide breakdown and lysosomal enzyme secretion induced by fMet-Leu-Phe in rabbit neutrophils. Likewise, fMet Leu-Phe- or leukotriene B4-evoked [3H]inositol trisphosphate accumulation was inhibited by the toxin. These findings, taken together with evidence that pertussis toxin specifically causes inactivation of the guanine nucleotide binding protein (Ni), suggests that guanine nucleotide binding proteins may mediate coupling between calcium-mobilising receptors and phospholipase C mediated reactions in rabbit neutrophils. PMID- 2985433 TI - Separation of kappa-opioid receptor subtype from frog brain. AB - Complete separation of the [3H]ethylketocyclazocine [( 3H]EKC) specific binding (kappa subtype) from tritiated Tyr-D-Ala2-Me-Phe4-Gly-ol5 enkephalin (DAGO) and Tyr-D-Ala2-L-Leu5-enkephalin (DALA) binding (mu-and delta-subtypes, respectively) was achieved by Sepharose-6B chromatography and sucrose density gradient centrifugation of digitonin solubilized frog brain membranes. The apparent sedimentation coefficient (s20.w) for the kappa receptor-detergent complex was 13.1 S and the corresponding Stokes radius 64 A. The isolated fractions exhibited high affinity for EKC and bremazocine, whereas mu- and delta-specific ligands were unable to compete for the [3H]EKC binding sites, indicating that the kappa subtype represents a separate molecular to compete for the [3H]EKC binding sites, indicating that the kappa subtype represents a separate molecular entity from the mu and delta receptor sites. PMID- 2985434 TI - Possible involvement of two proteins (phosphoprotein and CD9 (p24)) in regulation of platelet calcium fluxes. AB - The monoclonal antibody ALB6 directed against the leukocyte differentiation antigen CD9 (p24) increases the calcium incorporation into isolated platelet membrane vesicles enriched in internal membranes. The similarities of the effects of both the monoclonal antibody and the catalytic subunit of the cAMP-dependent protein kinase (C, subunit), which phosphorylates a protein of an apparent molecular mass of 23 kDa, led us to investigate the relationship between CD9 (p24) and the 23-kDa phosphoprotein (p23). ALB6IgG does not inhibit the C.subunit induced phosphorylation of p23 and the immunoadsorption by ALB6IgG of p24 associated to membrane vesicles does not alter the phosphorylation pattern. Thus, proteins of similar molecular mass appear to be involved in calcium fluxes: one is recognized by the ALB6 antibody while the other can be phosphorylated by the C subunit. PMID- 2985436 TI - Cloning and sequencing of the Pseudomonas aeruginosa PAK pilin gene. AB - A 1.2-kilobase (kb) HindIII restriction fragment containing the pilin gene from Pseudomonas aeruginosa PAK has been cloned and sequenced. The pilin protein is 144 amino acids in length with a positively charged leader sequence of 6 amino acids. There is probably only one copy of the gene per chromosome. PMID- 2985435 TI - A second human calcitonin/CGRP gene. AB - The calcitonin (CT) gene is alternatively expressed in a tissue-specific fashion producing either the calcium regulatory hormone CT in the thyroid or the neuropeptide calcitonin gene related peptide (CGRP) in the brain. In medullary carcinoma of the thyroid both peptides are produced. We present here evidence for the existence in the human genome of a second CT gene, which is also expressed in human medullary thyroid carcinoma. This gene encodes a second human CGRP, differing from the known human CGRP in 3 of the 37 amino acids. PMID- 2985437 TI - Alkaline phosphatase of the calf intestine hydrolyzes phospholipids. AB - Pure alkaline phosphatase of the calf intestine is able to hydrolyze phosphatidylinositol 4,5-diphosphate (TPI) to phosphatidylinositol and Pi and to dephosphorylate phosphatidic acid. This phosphomonoesterase activity shows a considerably high specific activity when an incubation medium at neutral pH containing 3 mM deoxycholate is used. The activity is inhibited by low concentrations of Ca2+. The enzyme has no detectable phosphodiesterase activity under the conditions tested. PMID- 2985438 TI - Insulin release independent of a rise in cytosolic free Ca2+ by forskolin and phorbol ester. AB - The role of cytosolic free Ca2+ in insulin release was evaluated using isolated rat pancreatic islets permeabilized with digitonin and incubated in Ca-EGTA buffers to fix free Ca2+ concentration at arbitrary levels. Ca2+ induced insulin release in a concentration-dependent manner with the threshold being between 0.1 and 1 microM. The hormone release was increased by forskolin and 12-O tetradecanoyl phorbol-13-acetate (TPA), a potent activator of adenylate cyclase and that of protein kinase C, respectively. The findings suggest that activation of both protein kinase A and protein kinase C modulate insulin release without a concomitant increase in cytosolic free Ca2+. PMID- 2985439 TI - The platelet activation induced by wheat germ agglutinin. AB - In human platelets, wheat germ agglutinin (WGA) induced serotonin release without cell agglutination. WGA induced the phosphorylation of both 40-kDa and 20-kDa proteins in a parallel manner, and at least, the phosphorylation of 40-kDa protein was preceded by transient formation of endogenous diacylglycerol (DG) accompanied by a decrease in phosphatidylinositol (PI). Both phosphorylation of these two proteins and serotonin release were inhibited by prior treatment of platelets with dibutyryl cyclic AMP, W-7, or TMB-8. These results suggest that both phosphatidylinositol turnover and Ca2+ mobilization play an essential role in WGA-induced platelet activation. PMID- 2985440 TI - Comparative binding properties of linear and cyclic delta-selective enkephalin analogues: [3H]-[D-Thr2, Leu5] enkephalyl-Thr6 and [3H]-[D-Pen2, D-Pen5] enkephalin. AB - The range of delta-selectivity of linear and cyclic analogues of enkephalin in rat brain was found to be: [D-Pen2, L-Pen5] enkephalin (DPLPE) greater than [D Pen2, D-Pen5] enkephalin (DPDPE) greater than [D-Thr2, Leu5] enkephalyl-Thr6 (DTLET) greater than [D-Ser2, Leu5] enkephalyl-Thr6 (DSLET). Saturation experiments performed with [3H]DPDPE and [3H]DTLET in NG108-15 cells and rat brain showed similar binding capacities for both the ligands, but the delta affinity of [3H]DTLET (KD approximately 1.2 nM) was much better than that of [3H]DPDPE (KD approximately 7.2 nM). The rather low delta-affinity of DPDPE induced high experimental errors cancelling the benefit of its better delta selectivity. Binding experiments in rat or guinea-pig brains showed, in both cases, the better delta-selectivity of [3H]DTLET compared to [3H]DSLET. The former peptide remains at this time the most appropriate radioactive probe for binding studies of delta-receptor. PMID- 2985441 TI - Selective localization of calpain I (the low-Ca2+-requiring form of Ca2+ dependent cysteine proteinase) in B-cells of human pancreatic islets. AB - An immunohistochemical study was performed to localize two distinct Ca2+ proteases (low-Ca2+-requiring calpain I and high-Ca2+-requiring calpain II) and their specific inhibitor (calpastatin) in human pancreas using the respective monospecific antibodies. Strongly positive staining by anti-calpain I antibody was found in pancreatic islets, specifically in B-cells, whereas the exocrine pancreatic tissue showed essentially no positive immunostaining. No such specific staining was found with anti-calpain II antibodies or anti-calpastatin antibodies. The results suggest that the Ca2+-dependent proteolysis in B-cells can be triggered by a small rise of the intracellular Ca2+ concentration without serious interference by the endogenous inhibitor. PMID- 2985442 TI - Polymerization of G-actin by caldesmon. AB - Electron microscopy of negatively stained samples indicates that caldesmon induces polymerization of G-actin into filaments. Polymerization takes place in a very low ionic strength solution and is accompanied by an increase of intensity of fluorescence of G-actin labelled with N-(1-pyrenyl)iodoacetamide. The effect of caldesmon is abolished by calmodulin in the presence of Ca2+. PMID- 2985443 TI - Further characterization of cardiodigin, Na+, K+-ATPase inhibitor extracted from mammalian tissues. AB - This study was undertaken to characterize endogenous digitalis-like activity in water extract from mammalian tissues. Prepurified samples obtained from guinea pig heart were analysed by reverse-phase HPLC using an acetonitrile gradient. The eluent was assayed for its activity as inhibitor of human heart Na+, K+-ATPase and digoxin-like immunoreactivity. Both activities were recovered in the same fraction after two successive chromatographic steps. These results provide further evidence for the presence of an endogenous digitalis-like factor, cardiodigin, in mammalian heart. PMID- 2985444 TI - Degradation mediated OH radical generation from synthetic cyclic peroxides: ESR studies. AB - Hydroxy radical generation on thermal decomposition of synthetic cyclic peroxides was determined by ESR using DMPO as a spin-trapping reagent. Both the 6- and 7 membered cyclic peroxides, 4-alkoxy-2,3-benzo-dioxan-1-ols and 4-alkoxy-2,3 benzodioxepin-1-ols, respectively, which were synthesized according to our idea of a radical-releasing drug, gave rise to DMPO-OH signals on heat treatment, i.e. under their degradation conditions. The signals were completely abolished with higher concentrations of OH scavenger. PMID- 2985445 TI - Probing the adenosine receptor with adenosine and xanthine biotin conjugates. AB - Biotin-containing analogs of a potent agonist (N6-phenyladenosine) and a potent antagonist (1,3-dipropyl-8-phenylxanthine) of adenosine receptor activity have been synthesized. A spacer chain to the biotin moiety is attached in both cases to the para-position of the phenyl ring. Two biotin conjugates of N6 phenyladenosine differing only in the length of the spacer chain bind to the adenosine receptor and to avidin simultaneously. The shorter-chain derivative was more potent in inhibiting binding of N6-[3H]cyclohexyladenosine to rat cerebral cortical membranes (Ki of 11 nM in the absence of avidin, 36 nM for the avidin complex). Three biotin conjugates of 1,3-dipropyl-8-phenylxanthine bound competitively to the adenosine receptor, but only in the absence of avidin. The results are interpreted in terms of the possible orientation of the ligands at the receptor binding site. PMID- 2985446 TI - Association of newly synthesized pro-opiomelanocortin with secretory granule membranes in pituitary pars intermedia cells. AB - The prohormone, pro-opiomelanocortin (POMC) is synthesized on ribosomes, subsequently routed to the Golgi apparatus and finally packaged into secretory granules where it is processed to various biologically active hormones (alpha melanotropin, adrenocorticotropin, beta-endorphin and beta-lipotropin). We report here that in frog and mouse pars intermedia cells, newly synthesized [3H]Arg labeled POMC is associated with the secretory granule membrane prior to processing. This association with the secretory granule membrane may be related to the intracellular transport and packaging of POMC and/or the facilitation of processing of the prohormone within the organelle. PMID- 2985448 TI - Rapid in vitro desensitization of the testosterone response in rat Leydig cells by sub-active concentrations of porcine luteinizing hormone. AB - We have studied in rat Leydig cells, the effect of sub-active concentrations of porcine LH on the subsequent stimulation of the cAMP and testosterone production by a sub-maximal concentration of pLH or hCG. We found that extremely low concentrations of pLH (0.01-2.0 ng/ml) were able to induce rapidly a partial but highly significative desensitization of the testosterone response without affecting the cyclic AMP response. These data indicate that desensitization of the steroidogenic response might be due to some lesion beyond cAMP formation or at the level of one discrete compartment of cyclic AMP, directly involved in the control of steroidogenesis. Moreover, our data strongly suggest that the basal circulating concentrations of LH can exert an inhibitory control on the testosterone response to LH pulses in vivo. PMID- 2985447 TI - Resonance Raman evidence of chloride binding to the heme iron in myeloperoxidase. AB - The resonance Raman spectra of ferric derivatives of myeloperoxidase at pH 8 show ligand-dependent differences. The data are consistent with the resting enzyme and the chloride and fluoride derivatives all having 6-coordinated high-spin configurations. At pH 4 we find that the resting enzyme is susceptible to photodegradation from our low power incident laser beam. Chloride binding inhibits this denaturation. Our data support direct binding of chloride to the enzyme under physiological conditions. PMID- 2985449 TI - The quaternary structure of E. coli inorganic pyrophosphatase is not required for catalytic activity. PMID- 2985450 TI - Purification of protein phosphatase from hen oviduct. AB - Two protein phosphatases of 103 and 29 kDa as determined by gel filtration, were purified from hen oviducts. The 103 -kDa phosphatase was purified 7300-fold to near homogeneity and dissociated into two polypeptides in the presence of SDS. Molecular masses of these polypeptides were estimated to be 60 and 38 kDa by SDS polyacrylamide slab gel electrophoresis using the buffer system of Laemmli, but 68 and 35 kDa using the buffer system of Weber and Osborn. The stoichiometry of these polypeptides was approx 1:1 according to the densitometric analysis of gels at 550 nm. The 29 -kDa phosphatase was purified 2900-fold. Both phosphatases dephosphorylated the alpha-subunit of phosphorylase kinase more rapidly than the beta-subunit. PMID- 2985451 TI - Growth-related variation of alpha 2-adrenergic receptivity in the HT 29 adenocarcinoma cell-line from human colon. AB - The human colon adenocarcinoma cell-line HT 29 has been shown to possess functional alpha 2-adrenergic receptors. Here, [3H] clonidine was used as radioligand to study the evolution of alpha 2-adrenergic receptivity during the time course of HT 29 cell culture. Scatchard analysis of the saturation curves indicates that the number of [3H]clonidine binding sites increases throughout the 17 day culture period. The maximal number of alpha 2-adrenoceptors is found during the stationary phase of growth, when cell density is high and mitotic rate low. Moreover, the use of adrenaline and clonidine as alpha 2-adrenergic agonists reveals a relationship between the number of receptors and the intensity of the biological effect associated with their stimulation (inhibition of the VIP induced cyclic AMP accumulation). PMID- 2985452 TI - Treatment of metastatic breast cancer with aminoglutethimide after progression on chemotherapy and/or hormonal therapy with tamoxifen. AB - Sixty patients suffering from metastatic breast cancer, whose disease progressed on chemotherapy and/or hormonal therapy with tamoxifen, were treated with aminoglutethimide. The overall objective response rate was 33%, including 3% complete responses. The median duration of response was 8 months. Even patients who failed to respond to prior treatment modalities responded objectively to aminoglutethimide. Objective response was observed in all metastatic sites, except for lung. Subjective improvement was achieved in 70% of the patients. Although initial toxicity was high (67%), side effects of aminoglutethimide were transient, and therapy had to be discontinued in only 3 patients. The results of this study indicate that aminoglutethimide in combination with hydrocortisone is a very effective treatment for metastatic breast cancer, especially in patients with positive hormone receptors. PMID- 2985453 TI - Carcinomatous meningitis associated with infiltrating lobular carcinoma of the breast. AB - The records of 365 patients with advanced carcinoma of the breast were examined to identify those with CNS involvement. Nineteen (5.2%) developed parenchymal cerebral deposits and 10 (2.7%) developed meningeal infiltration during the course of their disease. Parenchymal cerebral deposits were almost exclusively associated with infiltrating duct carcinoma (95%) and meningeal infiltration (carcinomatous meningitis) was almost exclusively associated with infiltrating lobular carcinoma. Meningeal infiltration occurred late in the course of advanced disease and was associated with diffuse involvement of the bone marrow and abdominal structures. PMID- 2985454 TI - Mechanisms of seizure control mediated by gamma-aminobutyric acid: role of the substantia nigra. AB - The substantia nigra has been identified as a critical site at which gamma aminobutyric acid (GABA) agonist drugs act to reduce susceptibility to a number of types of experimentally induced generalized seizures. Moreover, the ability of systemically administered GABA-elevating agents to protect against maximal electroshock seizures is directly correlated with an increase in GABA specifically in the nerve-terminal compartment of substantia nigra. The significance of these findings is discussed in terms of the role of specific nigral synapses for the control of seizure propagation. Evidence from lesion studies, as well as studies with opiates and substance P analogs, further supports the hypothesis that Inhibition of nigral efferents reduces susceptibility to generalized seizures. Inhibition of nigral outflow causes a decreased sensitivity to chemoconvulsants without precluding the animal's ability to exhibit any or all of the motor components of a seizure. We therefore propose that nigral outputs are capable of facilitating seizure propagation and can function as a gating mechanism for the generalization of convulsive activity. PMID- 2985455 TI - Gluconeogenesis in the chicken: regulation of phosphoenolpyruvate carboxykinase gene expression. AB - In chickens, the liver functions in gluconeogenesis to recycle lactate carbon (Cori cycle) and the kidney is the major organ for net gluconeogenesis from substrates such as pyruvate and amino acids. This is markedly different from mammalian systems where the liver is the primary gluconeogenic organ. The limited ability of chicken hepatocytes to synthesize glucose is explained, at least in part, by the observation that phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) in these cells is located exclusively in the mitochondria. The kidney possesses a cytosolic form of this enzyme that adapts to dietary and acid-base stimuli. The relative abundance of mRNA coding for the cytosolic enzyme has been detected by using a specific cDNA probe. Starvation increases the level of this mRNA in chicken kidney and also results in the appearance of the message in chicken liver. Isolated hepatocytes have been used to determine which hormones regulate expression of the hepatic gene. Incubations with glucagon, epinephrine, norepinephrine, dexamethasone, or dibutyryl cyclic AMP increase the relative abundance of the message in liver cells isolated from fed chickens. Despite considerable levels of this mRNA in the liver of starved chickens, functional cytosolic enzyme activity is not detected. This indicates some form of posttranscriptional regulation. The studies summarized illustrate the usefulness of isolated hepatocytes and recombinant DNA probes in the study of hormonal regulation of hepatic gene expression. PMID- 2985456 TI - [Cyclic AMP content of the blood, brain, stomach, adrenals and duodenum after administration of neurotensin and cholecystokinin]. PMID- 2985457 TI - Development of the major pathways for neurite outgrowth in the chick hindlimb. AB - To elucidate mechanisms that may control development of the gross anatomical nerve pattern, motoneuron outgrowth into the chick hindlimb was examined using orthograde labeling, scanning and transmission electron microscopy, and Alcian blue staining. Results show that growth cones are not guided by contact with oriented extracellular fibrils, aligned mesenchyme cells, the myotome, or the vasculature. Pathways are not delineated by cell-free space or channels of lower cell density; however, densely packed mesenchyme may form barriers that channel outgrowth. In addition, abundant mesenchymal cell death was seen at the nerve front. This cell death may provide space that encourages growth cone advancement. Pathways often lie along interfaces between areas that stain darkly and lightly with Alcian blue, which specifically stains glycosaminoglycans, and growth cones never penetrate areas that stain intensely, such as the pelvic girdle, which is known to be a barrier to outgrowth. Leading growth cones form specialized contacts with mesenchyme cells, but the predominant contacts are interneuronal. It is proposed that the anatomical pattern of outgrowth is determined by the distribution of preferred substrata, the most preferred substratum being other neurites. Further, neurites tend to prefer loose mesenchyme to dense mesenchyme or areas rich in glycosaminoglycans. PMID- 2985458 TI - The growth of motor axons in the spinal cord of Xenopus embryos. AB - The innervation of the myotomal muscles in the trunk region of Xenopus embryos has been examined to see how the path taken by motoneurons within the spinal cord is formed. The growth of motor axons has been studied by retrograde labeling with horseradish peroxidase and the growth of the spinal cord and myotomes has been studied by labeling with fluorescent beads. Results show that motoneurons initially innervate the nearest muscles. Then through a process of differential growth whereby the muscles elongate more than the spinal cord, the axonal terminals in the muscles become displaced caudally relative to their cell bodies. In this manner the central pathway taken by the motor axons develops after initial innervation of their peripheral targets. PMID- 2985459 TI - Correlations between in vitro and in vivo mechanisms of pyrethroid insecticide action. AB - Classical neuropharmacological procedures have been used to elucidate insecticide mode of action in vitro. A good deal has thus been learned about pyrethroids but novel techniques have been necessary to explain the toxicology and symptomology. The free-walking, electrode-implanted cockroach technique, which was developed for this purpose, is described. It enabled correlations to be made between symptomology and effects on specific nerves. The negative temperature coefficient of toxicity of allethrin (the first pyrethroid) was explained in terms of repetitive firing in peripheral (sensory) nerves rather than by nerve blockage, which had been suggested from previous in vitro studies. The elucidation of target sites in vivo and the most useful parameter to study, i.e., repetitive firing in nerve axons, enabled the definition of a pyrethroid resistance mechanism in a major insect pest. It also showed two modes of action for permethrin and cypermethrin, its alpha-cyano analog. A structure-activity relationship for a range of pyrethroids, combining in vitro and in vivo approaches, confirmed two distinct types of pyrethroid action. Studies of poisoning signs and nerve disruptions in vivo in the mouse and cockroach, using diazepam in conjunction with pyrethroids, implicated the GABA-receptor complex as a target for alpha-CN-phenoxybenzyl pyrethroids. This was confirmed by making conductance measurements in crayfish claw opener muscle fibers in vitro. PMID- 2985461 TI - Neuropharmacologic and neuropathologic effect of fenvalerate in mice and rats. AB - B6C3F1 mice and Sprague-Dawley rats displayed the characteristic signs of pyrethroid intoxication following single oral doses ranging from 56 to 320 and 133 to 1000 mg/kg fenvalerate, respectively. The LD50s for mice and rats were 180 and 776 mg/kg, respectively, with corn oil as the vehicle. Signs of neurologic deficit such as splayed gait, tremors, ataxia, and hind limb incoordination were observed at doses of greater than or equal to 100 mg/kg (mice) and greater than or equal to 133 mg/kg (rats) within 1-8 hr after dosing. These signs had disappeared in most animals within 72 hr. Slight peripheral nerve fiber damage was detected in surviving mice and rats sacrificed 10 days after dosing. The incidence and severity were dose related at doses greater than or equal to 56 and greater than or equal to 180 mg/kg; however, even at lethal doses, evidence was lacking for the presence of nerve lesions in several animals. Thus two distinct neurologic effects were observed: a reversible ataxia/incoordination and a neuropathologic effect manifested as sparse axonal damage in peripheral nerve. PMID- 2985460 TI - Cellular sources of fibrotic collagen. AB - Every somatic cell within vertebrate species contains genetic information and therefore the potential to synthesize collagen. Normally, when there is a requirement for new connective tissue following injury, cells of mesodermal origin are activated to meet that demand and supply sufficient amounts of collagen to return structure and function to the damaged tissue or organ. Under these conditions, nonconnective tissue cells either do not synthesize collagen or may produce only minute quantities which are barely detectable by our most sensitive techniques. However, when these nonmesodermal cells are exposed to an abnormal or toxic environment, collagen expression may become greatly enhanced. For example, when hepatocytes are removed from their normal in vivo environment and placed in cell culture, they demonstrate a dramatic increase in their potential to produce collagen. Hepatocytes may also express this ability to produce significant quantities of collagen when they are exposed to toxins or injury in vivo. The list of cells shown to produce collagen or contain enzymes related to collagen biosynthesis has expanded in recent years and now includes such diverse and specialized cells as glomerular mesangial, adipocytes, aortic endothelial, smooth muscle, Schwann cells, and lymphocytes. Therefore, the loss of structure and function in a tissue or organ due to fibrosis may result from excessive deposition of collagen from nonmesodermal sources as well as from connective tissue cells. PMID- 2985463 TI - Corticotrophin-like bioactivity in the pituitary gland and brain of the pacific hagfish, Eptatretus stouti. AB - The cytochemical bioassay for corticotrophin (ACTH) was used in an attempt to detect ACTH-like activity in the Pacific hagfish, Eptatretus stouti. Extracts of the pituitary gland and brain were active in this assay system but those of liver and skeletal muscles were not. The slopes of the dose-response lines of the pituitary extracts were less than those of the mammalian corticotrophin standard preparation but greater than those of the brain extracts. The results suggest that the corticotrophic factor in the hagfish differs from mammalian ACTH. PMID- 2985462 TI - Chronic inhalation of short asbestos fibers. AB - An animal inhalation study was initiated to study the chronic biological effects of inhalation of short chrysotile asbestos fibers. Rats and monkeys were exposed for 18 months, 7 hr/day, 5 days/week to a specially prepared, chrysotile asbestos aerosol. Based upon daily chamber measurements, the mean concentration of fibers in the chamber air was 1.0 mg/m3. By phase contrast microscopy, the number of fibers greater than 5 micron in length was determined to be 0.79 fiber per cubic centimeter. Rats were autopsied for pathological and histochemical examination at 1, 3, 6, 12, 18, and 24 months after initiating exposures. No significant differences in the histochemical data were seen between the exposed and control groups. Gross and histopathologic examination of exposed and control groups of rats indicated no compound-related lesions, including fibrosis. Open lung biopsies were performed on the chrysotile-exposed and the control monkeys 28 months after initiating exposures. Histopathologic evaluation of the lung biopsy tissue showed the presence of asbestos bodies adjacent to the terminal bronchioles of the asbestos-exposed monkeys. There was no observed fibrosis in pulmonary tissue. All monkeys are being maintained for an indefinite period and observed for signs of latent pulmonary disease. PMID- 2985464 TI - 3,5,3'-Triiodothyronine receptors and thyroxine 5'-monodeiodinating activity in thyroid hormone-insensitive amphibia. AB - Adult anuran Amphibia and neotenous urodeles, such as Necturus maculosus, rarely respond to thyroid hormone (TH). In this study, the possibility was examined that this lack of response is due either to an inability to convert thyroxine (T4) to 3,5,3'-triiodothyronine (T3) or to the absence of nuclear TH receptors. Following injection of [125I]T4, significant amounts of [125I]T3, analyzed by chromatography, were detected in the serum and liver of Necturus and Rana catesbeiana frog indicating that both possesses a T4 5'-monodeiodinating system. Nuclear binding of T3 was studied in suspensions of purified hepatic nuclei and intact red blood cells (RBC). Analysis of binding data revealed that frog liver nuclei contained two sets of saturable T3 binding sets with affinities comparable to those of the two sets of sites previously demonstrated in tadpole liver nuclei (Galton and Schaafsma, 1983). However, the number of sites per nuclei was small compared to tadpole; expressed as pmol/mg DNA, the maximum binding capacity of the high affinity set of sites (MBC1) was 2.2 +/- 0.31 versus 12.9 +/- 1.80 and MBC2 was 24.9 +/- 4.5 versus 42.2 +/- 54. Receptor number in RBC nuclei was also smaller in frog than tadpole: 90 +/- 26 versus 882 +/- 56 sites/nucleus (Kd less than 10(-11) M in both groups). No comparable high affinity binding sites were detected in Necturus liver, but some sites were found in Necturus RBC. These cells contained 2111 +/- 120 sites/nucleus, more than twice the number found in tadpole.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985465 TI - Human chorionic gonadotrophin and immature fish ovary: characterization and mechanism of the in vitro stimulation of cyclic adenosine monophosphate accumulation. AB - Human chorionic gonadotrophin (hCG) increased cyclic adenosine monophosphate (cAMP) in pieces of eel ovary in vitro. Although the ED50 (about 0.1 micrograms/ml) was the same as for carp gonadotrophin (cGTH), the maximal stimulation (close to 3 times basal level) was about 10 times lower than that for the fish hormone. The mechanism of this atypical action of hCG was studied using different approaches. Human CG did not inhibit the action of cGTH. Desensitization experiments (cGTH or hCG being injected in vivo 18 hr before the sampling of the ovary) were carried out; the in vitro action of hCG was suppressed by both pretreatments; the in vitro action of cGTH was largely reduced by in vivo cGTH and to a much lesser extent by in vivo hCG. Together with other data the results indicated that hCG acts via binding to adenylate cyclase-coupled receptors which also recognize cGTH; moreover, they suggested that among those systems only a fraction can be activated by hCG. We propose that in teleosts the pool of GTH receptors is not homogeneous but contains classes of "isoreceptors" differing by their recognition specificity at least. Parallel studies with ovine lutrophin (oLH) indicated that this hormone binds to the same adenylate cyclase coupled receptors which are sensitive to hCG but that it does it with a much lower affinity. Other characteristics of oLH/hCG-stimulated receptor-adenylate cyclase systems include an important release of cAMP into the incubation medium. The precise physiological significance of GTH "isoreceptors" still requires clarification. The ability of hCG to induce desensitization must be considered in fish culture experiments using this hormone. PMID- 2985466 TI - Effects of prolactin, adrenocorticotrophin, neurohypophysial peptides, cortisol, and androgens on some osmoregulatory parameters of the hypophysectomized catfish, Heteropneustes fossilis (bloch). AB - Daily injections of ovine prolactin (PRL), cortisol acetate (FA), arginine vasotocin (AVT), and adrenocorticotrophic hormone (ACTH), at doses ranging from 0.1 to 500 mU/g, 0.2 to 25 micrograms/g, 0.01 to 0.02 micrograms/g, and 5 to 10 mU/g respectively, for 5 days elevated plasma osmolarity and plasma sodium levels in the 3-day hypophysectomized catfish, Heteropneustes fossilis maintained in tap water. PRL enhanced urine production and decreased urine osmolarity and sodium concentration. Administration of FA and AVT increased urine output as well as urine osmolarity and sodium concentration, thereby resulting in severe natriuresis. Simultaneous administration of PRL and FA to hypophysectomized catfish restored plasma osmolarity and sodium concentration to normal levels even in the presence of increased urinary salt loss. However, if the fishes were maintained in deionized water, administration of PRL and FA had no effect on plasma osmolarity or plasma sodium levels suggesting that these hormones increase plasma osmotic pressure by stimulating active uptake of salts from the external medium. Neither PRL nor AVT evoked any increase in plasma cortisol level indicating that their effects on catfish osmoregulation are not mediated through cortisol production. Isotocin, testosterone, and delta 4-androstenedione had no effect on any of the plasmatic or urinary parameters. It is concluded that prolactin, cortisol, and AVT are the principal hormones for osmoionic homeostasis in this catfish and that they act through independent mechanisms. PMID- 2985467 TI - Immersion of rainbow trout in 3,5,3'-triiodo-L-thyronine (T3): effects on plasma T3 levels and hepatic nuclear T3 binding. AB - An immersion protocol was established for chronically administering thyroid hormone to fish. Rainbow trout were immersed in solutions of 3,5,3'-triiodo-L thyronine (T3) at 11 degrees C for various periods. Most plasma T3 adjustment occurred by 24 hr and within 5 days consistent plasma T3 levels were observed. Availability of unoccupied hepatic nuclear T3-binding sites was assessed in two experiments by uptake of nuclear radioactivity in T3-immersed trout injected 12 hr previously with [125I]T3. At low ambient T3 levels over 92% of nuclear radioactivity was identified chromatographically and immunologically as T3, but at high ambient levels (25 micrograms/100 ml) biliary [125I]T3 metabolites may contaminate the nuclear fraction. Displacement of [125I]T3 from nuclei was first detected at 0.25 micrograms T3/100 ml water, corresponding to a plasma T3 level of 0.55 ng/ml; maximum displacement, depending on experiment, occurred at 1.25 and 3.8 micrograms T3/100 ml, corresponding to plasma T3 levels of 14 and 9 ng/ml. In conclusion, chronic physiologic T3 treatment of these trout can be achieved by immersion in T3 solutions of 0.1-5 micrograms T3/100 ml water. PMID- 2985468 TI - [Chromosomal inversion accompanied by an enhancement of uridine phosphorylase gene expression in Escherichia coli K-12]. AB - In thymine requiring auxotrophs of Escherichia coli the uridine phosphorylase enzyme (udp gene) can catalyze nonspecifically conversion of thymine to thymidine. By selection for effective utilization of exogenous thymine, it is possible to isolate forms with increased expression of the udp gene. Mutants with increased gene expression were isolated from the strain with transposon Tn10 within the metE gene closely linked to udp. Some mutants (designated udpPf) losing Tn10 but retaining the Met- phenotype are characterized by disturbance of recombination in the metE-udp region: they do not form Met+ transductants in P1 transduction with the wild-type donor strain. However, recovery of homology in the chromosomal metE-udp region takes place with low frequency in P1 transduction using the strain with Tn10 insertion in metE as a donor. Data obtained in transductional and conjugational experiments demonstrate that the udpPf1 mutant studied is an inversion extending about 3 min of the E. coli chromosome and including the region of chromosomal replication origin (oriC). PMID- 2985469 TI - Recognition sequences of restriction endonucleases and methylases--a review. AB - The properties and sources of all known endonucleases and methylases acting site specifically on DNA are listed. The enzymes are crossindexed (Table I), classified according to homologies within their recognition sequences (Table II), and characterized within Table II by the cleavage and methylation positions, the number of recognition sites on the DNA of the bacteriophages lambda, phi X174 and M13mp7, the viruses Ad2 and SV40, the plasmids pBR322 and pBR328 and the microorganisms from which they originate. Other tabulated properties of the restriction endonucleases include relaxed specificities (Table III), the structure of the restriction fragment ends (Table IV), and the sensitivity to different kinds of DNA methylation (Table V). Table VI classifies the methylases according to the nature of the methylated base(s) within their recognition sequences. This table also comprises those restriction endonucleases, which are known to be inhibited by the modified nucleotides. Furthermore, this review includes a restriction map of bacteriophage lambda DNA based on sequence data. Table VII lists the exact nucleotide positions of the cleavage sites, the length of the generated fragments ordered according to size, and the effects of the Escherichia coli dam- and dcmI-coded methylases M X Eco dam and M X Eco dcmI on the particular recognition sites. PMID- 2985471 TI - [Method of detecting Enteroviruses in reservoir water using natural sorbents]. PMID- 2985470 TI - Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. AB - Three kinds of improvements have been introduced into the M13-based cloning systems. (1) New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors. Mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences. A new suppressorless strain facilitates the cloning of selected recombinants. (2) The complete nucleotide sequences of the M13mp and pUC vectors have been compiled from a number of sources, including the sequencing of selected segments. The M13mp18 sequence is revised to include the G-to-T substitution in its gene II at position 6 125 bp (in M13) or 6967 bp in M13mp18. (3) M13 clones suitable for sequencing have been obtained by a new method of generating unidirectional progressive deletions from the polycloning site using exonucleases HI and VII. PMID- 2985472 TI - [Changes in the functional state of neutrophils in rats exposed to noise]. PMID- 2985473 TI - [Body functions of operators of various types of plasma units]. PMID- 2985474 TI - Placental-site trophoblastic tumor (trophoblastic pseudotumor): case report demonstrating failure of chemotherapy, surgery, and radiotherapy to control metastatic disease. AB - A case of metastatic placental-site trophoblastic tumor (trophoblastic pseudotumor) is presented and compared to four previously reported cases. Chemotherapy with MAC (methotrexate, dactinomycin, cyclophosphamide), aggressive surgical debulking, and radiotherapy were ineffective in producing disease regression. Only two of the four similar cases in the literature showed short responses (4-7 months) to several different aggressive multiagent chemotherapy regimens. Optimal therapeutic outcome is likely to result from an early diagnosis and aggressive surgical treatment of localized tumor. PMID- 2985475 TI - Carcinoma of the cervix stage IV treatment problems. AB - Sixty-seven patients with cervical cancer stage IV were treated in 1967-72. Only 21 received a complete course of radiotherapy and none received surgery or chemotherapy. Two patients had mobile tumors with extension to the bladder. Both died from distant metastases without local recurrences. Two patients with metastases to the lymph nodes in the groin, died within 3 and 4 years, respectively. All the other patients with distant metastases survived less than 2 years. PMID- 2985476 TI - Endometrial "sarcomas" complicating ovarian thecoma, polycystic ovarian disease and estrogen therapy. AB - Unopposed endogenous and exogenous estrogenic stimulation has been considered by most investigators to have a role in the pathogenesis of carcinoma of the endometrium. Although a few cases of "sarcomas" of the endometrium that had developed in an estrogenic setting have been reported, a clear-cut association between estrogenic stimulation and these forms of endometrial cancer has not been established. We report six cases of endometrial sarcomas complicating ovarian thecomas, polycystic ovarian disease, or prolonged estrogen therapy. Three ovarian thecomas, which are considered to be estrogenic tumors, were associated with endometrial malignant mullerian mixed tumor, mullerian adenosarcoma, and low grade stromal sarcoma in postmenopausal women. Polycystic ovarian disease, a condition characterized by unopposed estrinism due to the peripheral conversion of excessive androstenedione to estrone, was found in a 27-year-old infertile woman with an endometrial malignant mullerian mixed tumor. A pure osteogenic sarcoma of endometrial stromal origin developed in a 28-year-old woman with gonadal dysgenesis (Turner's syndrome) who had received estrogens for 18 years. The sixth woman, with an empty sella turcica after radiation therapy of a pituitary adenoma, had an endometrial mullerian adenosarcoma at the age of 40 years after 16 years of estrogen therapy. None of these patients had had pelvic radiation therapy. The evidence from this series of cases and from six additional cases identified in the literature suggests that the risk of endometrial sarcomas may be increased by estrogen therapy or endogenous disorders that lead to unopposed estrogenic stimulation of the uterus. PMID- 2985477 TI - Determination of the intervillous perfusion index in pregnancies with intrauterine growth retardation. AB - The authors developed a new radioisotope technique to measure placental blood flow for early detection of placental insufficiency. Using this method placental perfusion has been measured in 20 healthy pregnant women and in 15 pregnancies complicated with intrauterine growth retardation (IUGR). The T-maximum pictures obtained made it possible to differentiate between the vascular and intervillous phases of placental blood flow. The time period of intervillous phase calculated as the percent of the whole placental T-maximum was given as the intervillous perfusion index (IPI). It has been demonstrated that IPI is significantly longer in IUGR pregnancies (67.0 +/- 14.6) than in the control group (31.6 +/- 10.7). These data suggest that the first sign of placental insufficiency is the prolongation of IPI, which is likely to precede the quantitative reduction of placental perfusion. PMID- 2985478 TI - [Bronchial cancer: diagnostic potentials and limits of chemotherapy. 1. Small cell bronchial cancer]. PMID- 2985479 TI - [Nuclear magnetic resonance in medicine--more than merely a new diagnostic imaging procedure]. PMID- 2985481 TI - [Experimental studies on lipoprotein metabolism in rats reared with liquid alcohol diet from the fetal life]. AB - Recently numerous reports show deleterious effects of alcohol abuse on pregnant women giving their children a high risk of stillbirth and/or several developmental abnormalities and mental retardation, i.e. the Fetal alcohol syndrome (FAS). In the present study, the effects of maternal alcohol consumption on lipid metabolism in the litter liver were investigated in rats. These rats showed not only quite less lipid deposition in spite of large amount of alcohol consumption up to adulthood, but also showed increased FFA oxidation in the livers. In addition, increased level of very low density lipoprotein and hypoglucagonemia were found. 40 micrograms/kg of glucagon which is known as an inhibitory factor of apoprotein production in the liver, was injected for 2 weeks into the rat tail vein and resulted in apparent fatty liver and hypolipoproteinemia. Norepinephrine injection (1 mg/kg) caused plasma glucagon to be depressed in the rat as compared with adult alcohol rats. Plasma cyclic AMP response to glucagon was also depressed in these rats. From these results, it is suggested that the deranged glucagon secretion from the pancreas and lowered glucagon-induced cyclic AMP response would relate to the abnormal lipoprotein metabolism in the rat. PMID- 2985480 TI - Primary hepatocellular carcinoma in the eastern province of Saudi Arabia: treatment with combination chemotherapy using 5-fluorouracil, Adriamycin and mitomycin-C. AB - An analysis is done of 40 evaluable patients treated with a combination of 5 Fluorouracil, adriamycin and mitomycin-C for primary hepatocellular carcinoma in Saudi Arabia. In only 5 patients (12.5%) objective partial remission was achieved. The duration of remission for responding patients was: 6, 6, 8, 12 and 20 weeks, respectively. Responders demonstrated a short survival time of 8, 8, 10, 15 and 33 weeks. The median survival time of non-responders was 8 weeks, which was not different from that of responders. An analysis of potential factors that might contribute to our poor results is presented. PMID- 2985482 TI - [The study of neuro-adrenolysis of pituitary gland on cancer pain and experimental approach to reveal its mechanism of pain relief]. AB - Neuro-adrenolysis of the pituitary gland by injection of pure alcohol was done to control intractable pain associated with wide-spread cancer, and pituitary functions were measured before and after the injection. Of a series of 46 patients undergoing a total of 57 neuro-adrenolysis of the pituitary gland, 22 (38.6%) of these procedures obtained complete pain relief 22 (38.6%) showed improvement, and the remaining 13 (22.8%) no effect. Hormonal changes of pituitary gland were not same among the cases and there was no correlation between hormonal changes and the occurrence of pain relief. Experimental study was done to investigate the effect of hypophysectomy and intracerebroventricular injection of AVP on pain threshold in rats. The study revealed that hypophysectomy and ventricular injection of AVP dose dependently raised pain threshold and these effects were inhibited by naloxone. These facts suggest that the analgesic effects of hypophysectomy and AVP injection into cerebral ventricle are mediated by beta-endorphin. PMID- 2985483 TI - Intraoperative sonography for brain tumor surgery. AB - Intraoperative ultrasonography was performed during craniotomies for nine cases of primary brain tumor. B-mode real-time linear scanner with a small probe head has proven to be extremely useful for detecting and localizing subcortical or deep seated brain tumors. The sonography clearly showed the tumors as hyperechogenic mass except in one case, calcifications as more echogenic particles, cyst as hypoechogenic area. Physical principles of echogenicity are discussed, and major advantages and disadvantages of intraoperative ultrasonography in neurosurgery are mentioned. PMID- 2985484 TI - Effects of continuous alpha (1-24) ACTH infusion in the dog. AB - Dogs chronically infused with alpha (1-24) ACTH for 2 weeks showed continuous elevations in plasma ACTH, cortisol, and progesterone levels. Haematologic changes included immediate increases in numbers of mature neutrophils and monocytes and reduced numbers of eosinophils and lymphocytes. Haematocrits were also reduced with ACTH infusion. Whereas serum potassium levels fell in association with ACTH, serum sodium was unchanged. Activities of two serum enzymes of probable liver origin, alkaline phosphatase and alanine aminotransferase, increased gradually with ACTH treatment. Histologic examination of liver tissue revealed prominent hepatocellular vacuolisation. The trophic action of ACTH infusion was manifested by an increased adrenal gland weight and an enhanced cortisol response to a bolus ACTH injection given 1 day after the infusion ceased. Long-term infusion of ACTH resulted in haematologic, biochemical and morphologic changes resembling those observed in dogs with spontaneous pituitary-dependent hyperadrenocorticism. PMID- 2985485 TI - Induction of glomerulopressin production by cyclic AMP. AB - Isolated rat livers were perfused with gassed Krebs-Ringer-Bicarbonate and different doses of theophylline and dibutyryl cyclic AMP were added to the perfusing solution. The perfusates were ultrafiltrated through Diaflo UM-05 membranes. The glomerulopressin activity of the ultrafiltrates were assayed in the tonic tension contraction (TTC) of isolated stomach fundus from rats. As glomerulopressin is known to be a glucuronide, it was inactivated with beta glucuronidase to confirm that the effect on the stomach fundus was due to the glomerulopressin and not to another substance. It was observed that doses of theophylline between 2 x 10(-3) M and 2 x 10(-5) M enhanced glomerulopressin production. However, there was no relationship between dose of theophylline and the response, and a dose of theophylline 2 x 10(-6) M has no activity. The perfusion with dibutyryl cyclic AMP at 5 x 10(-8) M increased the amount of glomerulopressin produced by the liver. This was a log-dose response of glomerulopressin production to dibutyryl cyclic AMP between 5 x 10(-8) M and 5 x 10(-4) M. Theophylline (2 x 10(-6) M) potentiated the activity of cyclic AMP (5 x 10(-8) M). These results support the view that cyclic AMP is intracellular mediator of the hepatic production of glomerulopressin. PMID- 2985486 TI - Opiate receptor blockade and diurnal pituitary and adrenal hormone levels. AB - Peripheral pituitary hormone levels exhibit circadian variations though the mechanism of these changes is unknown. In order to investigate the possible role of endogenous opiates in such changes we have studied the influence of opiate receptor blockade with naloxone (6.8 mg) on pituitary hormones in the morning and again in the evening in six normal male volunteers. Basal ACTH, cortisol, aldosterone and prolactin were higher in the morning than in the evening. Following naloxone at 0700h both ACTH and cortisol rose indicating a tonic inhibition of ACTH by endogenous opiates at that time. At 2230h cortisol rose following naloxone but ACTH did not, suggesting that endogenous opiates do not play an important role in the diurnal rhythm of this hormone and consistent with the suggestion that endogenous opiates can effect cortisol levels independently of their action on ACTH. Neither aldosterone nor prolactin were influenced by naloxone. In contrast TSH was unaffected by naloxone in the morning but fell in the evening (mean + SE decrement over 120 min -0.6 +/- 0.3 mU/l as compared with the control +0.6 +/- 0.4 mU/l; p less than 0.01). Thus, endogenous opiates probably tonically stimulates TSH levels in the evening when TSH may increase and possibly play a role in the circadian rhythm of TSH. PMID- 2985487 TI - Progesterone, 17-OH-progesterone, androstenedione and testosterone plasma levels in spermatic venous blood of normal men and varicocele patients. AB - Progesterone (P), 17-OH-progesterone (17-OH-P), Androstenedione (delta 4) and testosterone (T) plasma levels were measured in spermatic venous blood of twenty nine varicocele patients (V) and in twelve normal subjects (N). Our data reveal a significant decrease of the mean testosterone in the spermatic blood of varicocele patients with respect to normal controls: (N = 1708.7 +/- 223.8 (SEM) nmol/l, n = 10. V = 1190.9 +/- 101.1 (SEM) nmol/l, n = 29. P less than 0.03). An inverse correlation has been observed between the age of varicocele patients and 17-OH-P (n = 29. y = -33.38x + 1384.70, r = -0.59, P less than 0.01) and delta 4 values (n = 23, y = -1.62x + 85.65, r = -0.49, P less than 0.05). The 17-OH P/delta 4 ratio appears significantly augmented in varicocele patients with respect to normal controls (n = 4.80 +/- 0.86 (SEM), n = 12. V = 9.65 +/- 1.21 (SEM), n = 23.0.02 greater than P greater than 0.01). This indicates a deficiency in varicocele patients of 17-20 lyase activity. The positive correlation between the P/17-OH-P ratio and age of varicocele patients (n = 28, y = 0.007 x -0.090, r = 0.45, P less than 0.03) suggests a progressive impairment of 17-alpha hydroxylase in such patients as they grow relatively older. These data demonstrated that the reduced spermatic levels of testosterone in varicoceles are due to the enzymatic impairment of testosterone biosynthesis, concerning firstly 17-20 lyase activity and secondly 17-alpha-hydroxylase activity. The latter enzymatic impairment is age related as is seen from the significant increase of the P/17-OH-P ratio in older patients. PMID- 2985488 TI - Carcinoma of the breast with stromal multinucleated giant cells. AB - Three cases of invasive mammary carcinoma with stromal osteoclast-like giant cells are reported. All the specimens were grossly of reddish-brown colour. On microscopical examination two were invasive ductal carcinomas and the third was of mixed ductal and mucoid type. In the stroma all specimens showed numerous osteoclast-like giant cells, vascular proliferation, and accumulation of haemosiderin pigment. Supplementary immunohistochemical and electron microscopical investigations indicated that the giant cells originated from mononuclear stromal cells with only slight histiocytic properties. The giant cell formation was probably induced by the infiltrating carcinomatous tissue which is also believed to induce new blood vessel formation. However, there is no evidence of a direct relationship between these features. PMID- 2985489 TI - Liver cell dysplasia and hepatocellular carcinoma: a histological and immunohistochemical study. AB - Liver cell dysplasia (LCD) was found in 28 (60%) of 47 patients with hepatocellular carcinoma (HCC); 22 (79%) of them had associated liver cirrhosis. LCD was more frequently observed in posthepatitic cirrhosis (82%) than in the other forms. Carcinoembryonic antigen (CEA), alpha-1-antitrypsin (AAT) and alpha fetoprotein (AFP), as demonstrated by the peroxidase-antiperoxidase method, were similarly expressed both in normal and in dysplastic cells. Hepatitis B surface antigen was found in eight cases (17%), six of which were associated with LCD. HBsAg was rarely found in dysplastic cells and frequently displayed a peculiar perinuclear pattern. The possible preneoplastic role of LCD is stressed. PMID- 2985490 TI - Mapping parathyroid hormone, beta-globin, insulin, and LDH-A genes within the human chromosome 11 short arm by spot blotting sorted chromosomes. AB - Rearranged human chromosomes carrying segments of chromosome 11 were separated from the normal chromosome 11 by high-resolution chromosome sorting. Sorted chromosomes were tested with parathyroid hormone, beta-globin, insulin, and LDH-A gene-specific probes to determine the genes carried by each chromosome segment. Based on the gene content and karyotypes of these abnormal chromosomes, the parathyroid hormone, beta-globin, insulin, and LDH-A genes and the unique restriction fragment ADJ-762 are all located on the terminal band of the short arm of human chromosome 11 (band 11p15), with LDH-A proximal to the other loci. PMID- 2985491 TI - Localization by in situ hybridization of the coagulation factor IX gene and of two polymorphic DNA probes with respect to the fragile X site. AB - The coagulation factor IX gene and two other polymorphic loci corresponding to DNA probes 52 A and St 14 have been previously localized in the q27 to qter region of the human X chromosome. In order to study their localization with respect to the fragile site at Xq27-28, we have hybridized the three DNA probes to metaphase chromosomes of a boy with fragile X mental retardation. We show that probe 52 A is located in the proximal part of the Xq27 band, while the coagulation factor IX gene is on the distal part of this band, but proximal to the fragile site. The very polymorphic St 14 probe is located in the distal part of the Xq28 band, on the other side of the fragile site. PMID- 2985492 TI - DNA polymorphism in the 5' flanking region of the human carbonic anhydrase II gene on chromosome 8. AB - A restriction-fragment-length polymorphism (RFLP) is described which is associated with the human carbonic anhydrase II gene (CA2) that codes for one of the three genetically distinct carbonic anhydrase isozymes, CA I, CA II, and CA III. The isolated DNA was cleaved with several restriction enzymes and subjected to Southern blot hybridization analysis using a DNA probe containing the 5' end of the human CA II gene. A two allele RFLP which was detected with the restriction endonuclease, Taq I, is expressed phenotypically on Southern blots as either a 5.4 kilobase (kb) fragment or as 4.0 and 1.4kb fragments. These fragments result from the presence or absence of a Taq I recognition site in the 5' flanking region approximately 1.0kb from the initiation codon of the CA II gene. Segregation analysis showed that the alleles are inherited in a Mendelian fashion, with a frequency of 50%. PMID- 2985494 TI - Continuous in vitro production of IgA by a human colostral immortalized cell line. AB - Breast milk samples from 8 postpartum mothers were collected and incubated with Epstein-Barr virus, immortalizing their B-lymphocytes and giving rise to lymphoblastoid cell lines. Three samples showed no growing lymphocytes and five samples gave rise to transformed cell lines. IgA positive cells were selected from one such cell line by rosetting with anti-IgA-coated sheep erythrocytes. The emerging cell line was similarly reselected giving rise to an IgA-surface positive lymphoblastoid cell line. Monoclonal IgA-producing cell lines were obtained by cloning in the soft agar method. IgA is continuously secreted by the cell lines which have been growing in vitro for more than 24 mth. PMID- 2985493 TI - Isolation and characterisation of a cDNA clone for human apolipoprotein CI and assignment of the gene to chromosome 19. AB - We have synthesised a mixed oligonucleotide 17 bases long and used it to isolate cDNA clones for apolipoprotein CI (apo CI) from an adult liver cDNA library. The partial sequence of one of these clones confirms its identity. We have used this probe and Southern blotting techniques to identify the human apo CI gene in DNA from a series of rodent X human somatic cell hybrids. Our results provide evidence for the assignment of this gene to human chromosome 19. PMID- 2985496 TI - Thymocyte activating factors from SV40-transformed human embryo fibroblasts. AB - A novel factor was found in the medium conditioned by SV40-transformed human embryo fibroblasts, which stimulate concanavalin A-induced thymocyte DNA synthetic response. This activity was estimated to be 10-15 kD and divided into two activities by ion exchange chromatography. One of them is a protein molecule and the other is a glycoprotein. In addition, these activities are not derived from the growth factors reported previously such as interleukin 2 (Morgan, R., Ruscetti, F. and Gallo, R. C. (1976) Science 193, 1007-1008) and transforming growth factor (De Larco, J. E. and Todaro, G. J. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 4001-4005). PMID- 2985495 TI - Immunoglobulin C mu gene restriction fragment length polymorphisms associated with chronic renal failure. AB - The DNA's of 41 patients with various forms of renal disease and of 52 controls were investigated for restriction fragment length polymorphisms (RFLP), using a probe recognising the immunoglobulin Cmu heavy chain gene. With the restriction endonuclease Sst 1, 50 of the controls and 12 of the patients had the expected single 4.3 kilobase (kb) fragment. The remaining 29 patients and 2 controls displayed two patterns of banding, 8 patients and 1 control had a 6.8 kb band in addition to the 4.3 kb, and 21 patients and 1 control had a single band of 5.1 kb. In addition, a significant association between high creatinine levels (greater than 150 mumol/l) and abnormal bands was found (21/25 patients with high levels had abnormal bands compared with only 5/16 patients with normal levels). These results are evidence for an association between the human immunoglobulin heavy chain region and renal disease and they apparently confirm an association already reported at the protein level. However, the new RFLP bands, although reproducible and restricted to renal patients, occur in an area where few polymorphisms would be expected. Further, the association with high creatinine suggests some subtle interaction between the creatinine pathway and this area of the human chromosome. PMID- 2985497 TI - 5-Nucleotidase activity in isolated follicular dendritic cells. AB - Follicular dendritic cells isolated from mouse lymph nodes were incubated in the presence of AMP to test 5-nucleotidase (5-Nase) activity. Ultrastructural observations showed the presence of 5-Nase on external membranes but also some activity inside the nucleus. 5-Nase was found associated to Fc receptors labelled with homologous immunoglobulins fixed on colloidal gold particles. Lymphocytes and macrophages, found in association with the follicular dendritic cells, were either 5-Nase positive or negative. The hypothetical roles played by 5-Nase in germinal centers are discussed. PMID- 2985498 TI - Restriction fragment length polymorphism of C4 genes in mice with t chromosomes. AB - Genomic DNA was isolated from 29 t strains and 4 congenic lines of mice, digested with restriction endonucleases, and hybridized with a probe representing the complement component 4 (C4) gene. All but one of the enzymes revealed restriction fragment length polymorphism in this sample of C4-related genes. Double digestion analysis suggested the presence of three C4 gene copies in some of the t chromosomes and two copies in others. The enzymes distinguished 16 different haplotypes among the 33 strains tested. Based on their restriction fragment length patterns, the t strains could be divided into four groups with strains in each group more closely related to each other with respect to their C4-region genes than strains belonging to different groups. At least three of these four groups represent different branches of the evolutionary tree constructed for the t chromosomes. The C4-related genes of the chromosomes are in strong linkage disequilibrium with the class II genes of the H-2 complex. Typing for the Ss and Slp allotypes of C4 has revealed the presence of the Ss1 phenotype in two t strains and of the Slpa phenotype in one strain. PMID- 2985499 TI - Markers of viral hepatitis in pregnancy. PMID- 2985500 TI - Production of monoclonal antibodies to herpes simplex virus type 1 by hybridoma technique. PMID- 2985501 TI - Japanese encephalitis virus infection during pregnancy. PMID- 2985502 TI - Hepatitis A epidemic in Kerala state (India) in 1980. PMID- 2985503 TI - Tracheobronchial mucin receptor for Pseudomonas aeruginosa: predominance of amino sugars in binding sites. AB - Pseudomonas aeruginosa, a common respiratory tract colonizer and pathogen, adheres to injured tracheal cells and to tracheobronchial mucin. These phenomena suggest that there are specific receptors for this organism in the respiratory tract. The receptor on injured tracheal cells contains n-acetylneuraminic acid as the principal sugar, but the structure of the receptor in mucin has not been described. Using a microtiter plate assay to study bacterial adherence to mucin, we have partially characterized the mucin receptor for P. aeruginosa. The receptor for both nonmucoid and mucoid strains is sensitive to periodate oxidation, suggesting that it is carbohydrate in nature, and the amino sugars n acetylglucosamine and n-acetylneuraminic acid inhibited the adherence of both types of strains. Nonmucoid strains were more sensitive to inhibition by n acetylneuraminic acid than to inhibition by n-acetylglucosamine, but the mucoid strains varied in their sensitivities to inhibition by each amino sugar. Preincubation of mucin with heat-inactivated influenza A virus (which binds to neuraminic acid) significantly reduced the adherence of P. aeruginosa. Treatment of mucin with Clostridium perfringens neuraminidase also reduced bacterial adherence significantly. Treatment of mucin with pronase did not affect adherence. Our results suggest that n-acetylglucosamine and n-acetylneuraminic acid are important constituents of the binding sites for P. aeruginosa on human tracheobronchial mucin. PMID- 2985504 TI - Adaptation of Ehrlichia sennetsu to canine blood monocytes: preliminary structural and serological studies with cell culture-derived Ehrlichia sennetsu. AB - Ehrlichia sennetsu, the causative agent of human sennetsu rickettsiosis, was successfully propagated in primary canine blood monocyte cultures. The growth cycle of this organism appears to be similar to that of Ehrlichia canis. The antigen derived from our E. sennetsu cultures was used to develop an indirect fluorescent antibody test for detection and titration of serum antibodies to the organism. Using this test system, we found that five human serum samples obtained from patients clinically diagnosed as having sennetsu rickettsiosis were positive for anti-E. sennetsu antibodies. In addition, 29% of the serum samples obtained from 200 patients having a fever of unknown origin and residing in various regions of Malaysia were also serologically positive. All sera from apparently healthy individuals were negative in the test. Dogs inoculated with cell culture adapted E. sennetsu developed a significant specific antibody titer to E. sennetsu, and the organism was subsequently isolated from their blood. These animals showed no clinical evidence of disease. The possibility of a higher prevalence of human sennetsu rickettsiosis in Southeast Asia and the potential usefulness of the canine model for studies of human sennetsu rickettsiosis are discussed. PMID- 2985505 TI - Impairment of granulomatous inflammatory response to Histoplasma capsulatum by inhibitors of angiotensin-converting enzyme. AB - Systemic infection with Histoplasma capsulatum induced a granulomatous inflammatory response in the lymphoreticular organs of C57BL/6 mice that was associated with elevated levels of angiotensin-converting enzyme (ACE) in the spleens. To determine the influence of ACE on the granulomatous response, either captopril or MK 421, two inhibitors of ACE, were administered intraperitoneally to mice 6 h after intravenous injection of H. capsulatum and then daily for 1 week. Each ACE inhibitor sharply reduced ACE activity in the spleens of infected mice. Both drugs worsened the clinical severity of infection and significantly increased the growth of H. capsulatum in livers and spleens of mice infected for 1 week. The histopathological changes in mice given captopril were more severe, with massive infiltrates of macrophages in proximity to large aggregates of yeasts. Conversely, the administration of captopril for 2 weeks during the resolving phases of infection did not slow the healing of the granulomatous lesions, nor did it provoke a relapse of infection. Captopril did not promote the growth of H. capsulatum in artificial medium. This drug was not cytotoxic to peripheral blood leukocytes or to splenic leukocytes from normal and infected mice. Administration of captopril to normal mice for 1 week did not depress the response of splenocytes of concanavalin A or to phytohemagglutinin, nor did it diminish delayed-type hypersensitivity responses in vivo. Finally, captopril did not augment the growth of H. capsulatum within macrophages. Our results suggest that ACE may participate in the regulation of the granulomatous inflammatory response to H. capsulatum and that ACE inhibition impairs the protective effects of granulomatous inflammation during acute H. capsulatum infection. PMID- 2985506 TI - Inhibition of growth of Chlamydia trachomatis by human gamma interferon. AB - Treatment of HEp-2 cell cultures with highly purified human gamma interferon before infection resulted in the reduction of Chlamydia trachomatis (L2/434/Bu) infectious particle yield. Electron microscope studies showed that interferon did not affect chlamydial conversion to reticulate bodies but influenced the extent of maturation to elementary bodies. High interferon concentrations (greater than 350 IU/ml) inhibited inclusion body formation and resulted in the appearance of aberrant reticulate bodies. PMID- 2985507 TI - Symptoms and lung function in low-exposure to TDI by polyurethane foam manufacturing. AB - A cross-sectional investigation was performed in seven polyurethane foam manufacturing factories. Lung function tests were performed at the factories by spirometric and nitrogen wash-out methods. Sixty-seven persons were investigated before and after the work shift, and exposure to toluene diisocyanate (TDI) was evaluated by subject-carried sampling equipment on the same day. The day mean exposure to TDI was 0.008 mg/m3 (0.001 ppm). In the non-smokers there was an increased frequency of symptoms from the airways reported in the exposed subjects as compared to the unexposed controls. There were, however, no signs of acute or chronic effects on the lung function measured by spirometric and nitrogen wash out methods, which could not be explained by smoking. PMID- 2985508 TI - Distinctions between endemic and sporadic forms of Epstein-Barr virus-positive Burkitt's lymphoma. AB - Tumour cell lines were established in vitro from 16 cases of Epstein-Barr (EB) virus genome-positive Burkitt's lymphoma (BL), 7 of "endemic" origin (i.e. from holoendemic malarial areas of Africa and of New Guinea) and 9 of "sporadic" origin (i.e. from outside such high-incidence areas). All the BL cell lines thus established were monoclonal by immunoglobulin isotype expression and displayed a characteristic chromosomal translocation, t(8:14) or t(8:22), confirming their malignant origin. Clear differences observed between the individual BL cell lines appeared to be related to their endemic or sporadic status. All 7 endemic cell lines began growth as a carpet of single cells, often with small, loose clumps appearing in later passage. Whilst 3 lines of sporadic origin displayed a similar pattern to the above, the majority of sporadic lines grew as large, tight clumps of cells from the first passage onwards. These differences in growth pattern were reflected by differences in cell surface phenotype, as defined in indirect immunofluorescence tests using a panel of monoclonal antibodies (MAbs) specific for B-lineage-associated antigens. BL cell lines could be classified into 3 separate groups on the basis of their reactivity with 6 particular antibodies (MHM6, AC2, Ki-1, Ki-24, J5 and 38.13). All 7 endemic BL cell lines and 2 of the 3 sporadic BL cell lines which began growth as single cells showed a group-I cell surface phenotype (MHM6, AC2, Ki-1, Ki-24 negative; J5, 38.13 positive) in early passage. In contrast, all 6 sporadic BL cell lines which began growth in large clumps displayed a distinct group-II phenotype (MHM6, AC2, Ki-1 positive/negative; Ki-24, J5, 38.13 positive); in later passage most of these sporadic lines progressed to a group-III phenotype (MHM6, AC2, Ki-1, Ki-24 positive; J5, 38.13 negative) without loss of those immunoglobulin and chromosomal markers identifying the cells' malignant origin. These clear differences between endemic BL cell lines on the one hand and the majority of sporadic BL cell lines on the other suggest that endemic BL arises from a more restricted range of progenitor B cells than does the sporadic form of the disease. PMID- 2985509 TI - Papillomavirus infection of the lower genital tract: detection of viral DNA in gynecological swabs. AB - A total of 311 smears from the lower genital tract were examined by the filter in situ hybridization method to identify human papillomavirus (HPV) DNA. Of these 311 smears, 229 came from clinically and cytologically negative patients and served as a control group. In this group HPV-DNA was detected in 5 cases (2.2%). Of 82 cytologically positive cases (25 confirmed by histology) 56 (68%) contained HPV-DNA. A high prevalence of HPV 6/11 and absence of HPV 16/18 was found in cases with cytological signs of permissive HPV infection. In mild and moderate dysplasia all viruses occurred at almost the same frequency. In severe dysplasia/carcinoma in situ HPV 16/18 was found 5 times more frequently than HPV 6/11. HPV 16/18 was identified in all 4 invasive cancer cases. Cervical irrigation of colposcopically suspect areas was performed in 15 cytologically and HPV-DNA positive cases using the hydrodynamic filtration method. In 12 cases only the cells obtained from the colposcopically positive areas contained HPV-DNA. The sensitivity and reproducibility of the filter in situ hybridization was shown by: comparing the results obtained by HPV-DNA hybridization using Southern blot analysis of tumor biopsies; analysing the correlation of cytologic diagnosis and presence of HPV-DNA in follow-up examinations, and diagnosing presence or absence of HPV-DNA in parallel filters from the same patients. PMID- 2985510 TI - Spontaneous and induced primary oncogenesis in natural killer (NK)-cell-deficient beige mutant mice. AB - Spontaneous tumor development and primary oncogenesis were compared in a large number of NK4-deficient, homozygous C57Bl/6-bg/bg mice and their NK normal, heterozygous +/bg littermate controls. In a group of 167 retired breeders followed for spontaneous tumors, the probability of survival for mice eventually dying with a tumor was greater for the NK-competent, +/bg than for the NK deficient, homozygous C57BL/6-bg/bg mice (p = 0.0019), although the higher overall incidence of tumors in the bg/bg group (48%) was not significantly different from that in the +/bg group (37%). In the bg/bg group the incidence of tumor death appeared to increase relatively sharply in the 25- to 29-month age bracket compared to the fairly regular increase in incidence observed in the +/bg group. All the spontaneous tumors except 2 (discovered in +/bg mice) were classified histologically as widely disseminated malignant lymphomas. The other two were one squamous-cell carcinoma and one sarcoma. A total of 73 bg/bg mice injected s.c. with benzo[alpha]pyrene (BP) had a higher overall incidence of tumors (81%) (rhabdomyosarcomas) than 138 +/bg mice (64%) and in the largest group (0.3 mg, n = 85) the bg/bg group developed tumors, at a higher incidence (p = 0.01) and with a shorter latency (p = 0.025) than the +/bg group. On the other hand, mice injected with dimethylbenzanthracene or given 4 weekly doses of 160 rads of gamma irradiation showed no difference in overall tumor incidence. In addition, mice injected with various doses of DMBA-induced murine leukemia virus (DMBA-LV) also showed no significant difference in tumor incidence. Others have reported that some of these treatments (DMBA, split-dose irradiation) cause profound NK suppression, thereby reducing NK differences between the two groups of mice. These results suggest that a partial NK impairment in beige mutant mice early in life may lead to significantly greater rates of death with spontaneous malignant tumors late in life. Some primary oncogenesis treatments (BP) but not others (DMBA, split-dose irradiation, leukemia viruses) cause tumors with a greater incidence and shorter latency in beige mice. The results suggest, but do not prove, that NK cells play a role in surveillance against spontaneously arising, and possibly some types of carcinogen-induced, tumors. PMID- 2985511 TI - HSV- and chemical carcinogen-induced amplification of SV40 DNA sequences in transformed cells is cell-line-dependent. AB - Eleven simian virus 40-transformed cell lines from 5 different species were tested for their ability to amplify integrated simian virus 40 DNA upon infection with herpes simplex virus type I or treatment with various chemical carcinogens. Four cell lines were positive only for virus-induced gene amplification and two lines were positive for both carcinogen- and virus-induced gene amplification. Individual cell lines were assayed for the presence of an intact SV40 origin of replication, the expression of a functional SV40 T-antigen, and permissivity to herpes simplex virus replication. These parameters were found to be positive in all 6 amplification-competent cell lines. The ability of herpes simplex virus to amplify SV40 DNA sequences in transformed cells is greater than that of chemical carcinogens and can be suppressed by specific inhibitors of the herpes virus encoded DNA polymerase. PMID- 2985512 TI - Further study of beta-TGFs released by virally transformed and non-transformed cells. AB - Chicken embryo fibroblasts sensitized by ts RSV respond to TGFs present in the media of non-transformed FR3T3 and NRK-4 rat cells and of the same cells transformed by KiMSV or RSV. They also respond to TGFs present in the media of BHK hamster cells transformed by MoMSV, PyV or RSV. Two other indicator rat cell lines, untransformed NRK-4 and FR3T3, sensitized by ts KiMSV, respond to the same TGF-containing media, and this response is increased by exogenous EGF. Normal FR3T3 cells failed to respond to any of the media. The most sensitive target cells were the ts KiMSV-FR3T3 cells at the restrictive temperature (39.5 degrees C). All the media tested on NRK-49F target cells required EGF for their TGF activity which was essentially dependent on prior activation by acidification. These data show that the above media from non-transformed or transformed cells contain beta-TGFs, with no detectable accompanying alpha-TGF activity. The release of and the response to these TGFs are not interdependent. A function of ts mutant src and k-ras viral oncogenes, still expressed at the restrictive temperature, can sensitize non-responsive cells, without there being any specificity towards the TGF producer cells. PMID- 2985513 TI - The effects of aging after coronary arterial bypass grafting on the regulation of cardiac output during upright exercise. AB - We examined the effects of age on cardiac performance and the mechanisms that regulate cardiac output during upright exercise in patients free of myocardial ischemia after coronary revascularization. There were 90 subjects, aged 36 to 75 years, of whom 27 were greater than or equal to 60 years. There were no age related changes in resting heart rate, systolic blood pressure, left ventricular end-diastolic volume index, left ventricular end-systolic volume index, stroke volume index, cardiac index and left ventricular ejection fraction. There were, however, age-related changes in exercise capacity (y = 20 - 0.21x, r = -0.52, P less than 0.001); exercise heart rate (y = 185 - 1x, r = -0.42, P less than 0.001); exercise end-systolic volume index (y = 11 + 0.46x, r = 0.28, P less than 0.01) and exercise ejection fraction (y = 81 - 0.31x, r = -0.28, P less than 0.01). In a subgroup of 54 patients with comparable exercise workload (27 aged less than 60 and 27 greater than or equal to 60 years), the age-related differences in exercise end-systolic volume index, exercise ejection fraction and exercise cardiac index were not observed, but the exercise heart rate was still higher in the younger patients (y = 168 - 0.76x, r = -0.34, P less than 0.01). Thus, age modifies the compensatory mechanisms that regulate the cardiac output during exercise. Young and old patients alike show increases in end-diastolic volume and ejection fraction to maintain exercise cardiac output. The higher exercise heart rate in the younger subjects suggests a decrease in cardiac responsiveness to adrenergic stimulation associated with aging. PMID- 2985514 TI - Hypothalamic-pituitary-adrenal function in children with atopic dermatitis treated with clobetasone butyrate and its clinical evaluation. AB - Twenty-nine patients with symmetrical eczematous lesions were treated with 0.05% clobetasone butyrate cream. Clinical conditions and cortisol and ACTH concentrations were evaluated at the beginning and after 1 week in 12 children and after 2 weeks of treatment in 17 patients respectively. The lesions and symptoms improved steadily and no significant change in cortisol and ACTH plasma levels was observed throughout the study in all children. This steroid offers clinically effective topical antiinflammatory activity with a wide margin of safety. PMID- 2985515 TI - Basal cell carcinoma and breast carcinoma following repeated fluoroscopic examinations of the chest. AB - A 69-year-old white Italian woman was first seen at Memorial Sloan-Kettering Cancer Center in 1981 concerning several skin growths on her back. The patient had had several basal cell carcinomas surgically removed from her back during the preceding 5 years. There was no history of arsenic ingestion or prolonged sun exposure and her family history was negative for skin cancer. The patient had developed pulmonary tuberculosis in 1938 and was treated with pneumothorax therapy. She had had more than 50 fluoroscopic examinations of the chest following this therapy, as well as multiple diagnostic x-ray films since that time. She recalled that she had faced the fluoroscopy beam during the procedure. In 1959, she had a transabdominal hysterectomy for fibroid tumors. In 1980 she underwent a right modified radical mastectomy for adenoid cystic carcinoma of the breast; biopsies of lymph nodes were negative. Physical examination revealed a thin, white woman with a right mastectomy scar. On the back, clustered in the interscapular region, were multiple scars and nine erythematous nodules with pearly borders, telangiectasia, and translucent surfaces. Within several nodules there were areas of light and dark brown pigmentation. There were no other suspicious lesions on the head, chest, or extremities, nor did the patient show any evidence of the basal cell nevus syndrome. Biopsy of all lesions revealed basal cell carcinoma, some of which were pigmented, without evidence of chronic radiodermatitis. All lesions were treated with curettage and electrodesiccation three times with good cosmetic results (Fig. 1). PMID- 2985516 TI - Thalidomide therapy. An open trial. AB - In an open trial, 35 patients suffering from a variety of dermatologic conditions were treated with thalidomide. The drug was shown to be a valuable addition to the therapeutic armamentarium; however, the occurrence of a peripheral neuropathy will limit its use just as its teratogenic side effects have done for many years. PMID- 2985517 TI - Major colorectal cancer aetiological hypotheses do not explain mortality trends among Maori and non-Maori New Zealanders. AB - New Zealand colorectal cancer mortality rates are presented for the period 1947 1980. Mortality has been increasing and is now the highest in the world for both males and females in the age range 35-64; indeed New Zealand mortality rates for those aged 35-44 are approximately twice those of other countries with high mortality. By contrast, colorectal cancer mortality rates among Maoris, the indigenous New Zealanders of Polynesian descent, have been decreasing so that they are now less than half the non-Maori mortality rates. These findings cannot be explained by ethnic differences in consumption of the major proposed dietary risk factors: total fat, cholesterol, meat, fibre and beer. Possible differences in the prevalence of lactose malabsorption, in faecal mutagen activity, and in the prevalence of colorectal polyps warrant further investigation. PMID- 2985518 TI - Preparation and properties of tritiated human [Tyr18, Trp27]-beta-endorphin. AB - Tritiated [Tyr18, Trp27]-beta h-EP was prepared from the corresponding diiodotyrosine derivative by catalytic reduction in the presence of carrier free tritium gas. A photoaffinity probe for beta-endorphin (beta-EP) receptors was prepared by selective modification of [Tyr18, Trp27]-beta h-endorphin with 2 nitro-4-azidophenylsulfenyl chloride (2,4-NAPS-C1) under acidic conditions to yield [Trp18-2,4-NAPS-Trp27]-beta h-endorphin (NAPS-beta-EP). NAPS-beta-EP was purified by high performance liquid chromatography and characterized by ultraviolet absorption spectroscopy and peptide mapping. Tritiated NAPS-beta-EP was prepared from tritiated [Tyr18, Trp27]-beta h-endorphin with 2,4-NAPS-C1. The ability of NAPS-beta-EP to form covalent bonds to macromolecules due to photolysis was established using bovine serum albumin. The efficiency of photolytic cross-linking was 15% and the equilibrium dissociation constant was 1.3 X 10(-5) M. PMID- 2985519 TI - Adduct formation is involved in radiosensitization, mediated by cytochrome c, of phi X174 DNA by misonidazole. AB - Radiosensitization by misonidazole of biologically active phi X174 DNA, mediated by cytochrome c, is most probably at least partly due to formation of an adduct between sensitizer and DNA, which can be removed from the DNA by a mild alkaline treatment thereby restoring the activity of the DNA. PMID- 2985520 TI - Variability of the region of the herpes simplex virus type 1 genome yielding defective DNA: SmaI fragment polymorphism. AB - A SmaI map of class I defective DNA of herpes simplex virus type 1 (HSV1) was constructed using a set of deletion hybrid phages. Four SmaI fragments on the defective DNA had a variability in length common among 15 HSV1 isolates: the 1.45 kilobase (kb) fragment located within the BamHI-Z (map coordinates 0.936-0.949) fragment, the 0.92-kb fragment neighboring on the 'a' sequence, the 0.44-kb fragment containing the intervening sequence of immediate-early mRNA-5 gene, and the 0.205-kb fragment corresponding to the 'a' sequence. The four SmaI fragments have several sets of reiterated sequences, among which the 1.45- and 0.92-kb fragments hybridized with mammalian cellular DNAs (human, monkey, rabbit, and mouse). PMID- 2985521 TI - Molecular pathogenesis of equine coital exanthema: restriction endonuclease digestions of EHV-3 DNA and indications of a unique XbaI cleavage site. AB - Equine herpesvirus type 3 (EHV-3) DNA, isolated from purified virions of the large-plaque strain, was digested with the restriction endonucleases XbaI, Bg/II, EcoRI, and HindIII. Several lines of evidence indicated that the DNA extracted from purified virions was composed of long (L) and short (S) components and was present as two isomeric forms, P and IS. The evidence included: (i) after electrophoresis on agarose gels, the summed molecular weights of the digestion products exceeded that expected from intact, unit size DNA; (ii) quantitative measurements of radioactivity (molar ratios) indicated 'minor bands' (0.5 M) interspersed among the major (1.0 M) bands; and (iii) a brief digestion with lambda-5'-exonuclease, prior to digestion with restriction endonuclease, resulted in the loss of some submolar and molar ratio bands, indicative of three termini. A preliminary fragment linkage map of the XbaI digestion products revealed EHV-3 DNA to contain only one recognition site in the unique sequence of the S component. From this linkage map, the size of the S component was deduced to be (22.3 +/- 5) X 10(6) molecular weight. PMID- 2985522 TI - Induction of cytotoxic cell activity by peptide fragments of beta 2 microglobulin. AB - A heptapeptide fragment of beta 2-microglobulin has been described by Abiko in hemodialysates of uremic patients and found to inhibit E-rosette formation by human T cells. In order to study the possible immunoregulatory effects of this heptapeptide, we investigated its effect on human cytotoxic cell activity. Low concentrations (10(-8) to 10(-7)M) of the heptapeptide enhanced cytotoxic activity of human lymphocytes against herpes simplex virus-infected cells by 50 60% whereas higher concentrations (10(-4)M) depressed cytotoxicity. When lymphocytes were incubated with the heptapeptide during an in vitro sensitization assay, an even stronger enhancement was observed. In an attempt to define possible structure-activity correlations, synthetic peptide fragments were also tested for their effect on cytotoxic cell activity. Thus, the corresponding desHis-hexapeptide (2-7) had little enhancing effect on fresh lymphocyte cytotoxicity but significantly enhanced cytotoxic activity following presensitization. In contrast, further amino acid deletions resulting in 3-7, 4 7, 5-7 or 6-7 fragments had no significant activity. Alternatively, C-terminal deletions resulting in 1-6, 1-5 or 1-4 fragments also failed to exert any effect on lymphocyte-mediated cytotoxicity. We suggest that the heptapeptide and its 2-7 hexapeptide fragment may play a role in regulating host cytotoxic responses to viruses in health and disease. PMID- 2985523 TI - [What's new in American dermatology]. PMID- 2985524 TI - [Therapy of pemphigus. Critical remarks based on 44 clinical cases]. AB - The results are evaluated of therapy administered between 1957 and 1983 to 44 patients with pemphigus (28 pemphigus vulgaris, 5 pemphigus vegetans, 11 pemphigus seborrhoicus). The mean initial steroid dosage was 87 mg prednisone equivalent per day. The 5-year survival rate was 83%. Fifteen patients who were additionally treated with azathioprine during the initial phases of the disease had a 5-year survival rate of 100%. However, after having the disease for 5 years 41% of all pemphigus patients have to be rehospitalized. This percentage is only 21% in the group requiring less than 100 mg prednisone per day initially. Mortality related to pemphigus or infection occurs significantly earlier than cardiovascular deaths. The prognosis for survival is relatively good, but the prognosis for recovery remains uncertain. Combined therapy with corticosteroids and azathioprine in the early stages of the disease resulted in a 5-year survival rate of 100%, and is therefore recommended. However, even this therapeutic modality does not change the uncertain prognosis regarding recovery. PMID- 2985525 TI - Radioprotection of EMT6 tumor by a new class of radioprotectors based on a pseudo peptide cysteamine combination. AB - Although WR-2721 preferentially protects normal tissues against irradiation, it seemed desirable to find other drugs presenting a lower toxicity and the same radioprotective properties. A new compound, I 102, was selected; it was characterized on one hand by a coupling between cysteamine and an amino-acid, and on the other hand by an acetyl-group, which protects the thiol function. The effects of WR-2721 and of I 102 were studied on EMT6 tumors grafted on BALB/c mice. Whatever the size of the tumor, the cell survival increased as a function of the time elapsed between the injection of I 102 and the end of the irradiation (TI). In contrast, the radioprotection afforded by WR-2721 was found to be independent of TI. The survival curves suggest that, like WR-2721, I 102 protects essentially oxygenated cells. PMID- 2985526 TI - Isolation of cytopathic and noncytopathic bovine viral diarrhea virus from the spleen of cattle acutely and chronically affected with bovine viral diarrhea. AB - Both cytopathic and noncytopathic bovine viral diarrhea virus (BVDV) were isolated from 16 of 17 bovine spleens representing 11 herds that had experienced acute BVD and from 12 of 21 bovine spleens from 1 herd affected with chronic BVD. It was concluded that isolation of cytopathic and noncytopathic BVDV from the same spleen probably indicates that an animal with a persistent, noncytopathic BVDV infection was superinfected with a cytopathic BVDV. The prevalence (greater than 70%) of 2 viruses in the spleen of cattle with acute or chronic BVD suggested that persistent infection with noncytopathic BVDV may be an important factor in the pathogenesis of BVD. PMID- 2985527 TI - Prostaglandin dehydrogenase activity of rat and rabbit testicular tissues and accessory glands before and after castration. AB - Rat reproductive tissues contained both 15-hydroxy-prostaglandin dehydrogenase (PGDH) and delta 13-reductase activities, while rabbit tissues exhibited only PGDH activity. In the rat, delta 13-reductase activity was absent in those tissues that had a high specific activity of PGDH and was present in those tissues with a low specific activity. Total specific activity of PGDH was greatest in the testicular capsule, whereas total activity was greatest in the testicular parenchyma of both species. Total PGDH activity was highest in rat seminal vesicles, where it was second only to the testicular parenchyma. Castration significantly increased PGDH activity of the epididymis, prostate, and seminal vesicles of rats, while delta 13-reductase activity disappeared from those tissues. The specific activity of PGDH was greater in the rat than in the rabbit, except for the testicular parenchyma of the rabbit. The greater PGDH activity in rat testicular capsules, compared to those of the rabbit, correlated well with diminished contractility of the rat capsule. PGDH activity of the interstitial cells correlated well with the role of prostaglandins in androgen synthesis. PMID- 2985528 TI - Hexestrol residues and metabolites in the tissues of wethers injected with hexestrol dicaprylate or hexestrol. AB - Four young (23 kg body weight) and two mature wethers (52 and 92 kg body weight) were subcutaneously injected with hexestrol (HX) or HX dicaprylate (HX-D) and killed 41 d later. The HX residues, comprising free, glucuronide and KOH hydrolyzable forms plus metabolites were determined by gas chromatography after liquid-liquid extraction and silica gel chromatographic purification of tissue sample. The HX residues were observed to be at concentrations of .1 to 1.0 ppb in most of the tissues examined. Maturity of the animals and the two hormonal formulations resulted in little difference in residues. The KOH hydrolyzable fraction was hardly detected in the tissues examined. Free HX was a major residue in muscle, representing about 70% of HX residues. Glucuronide HX represented 70 to 80% of HX residues in liver and kidney. In lung, glucuronide and free HX were present in similar amounts. This study showed that the gross metabolic patterns of HX and its esters are similar to other estrogens and steroids. PMID- 2985529 TI - LHRH receptor, LH and FSH concentrations in anterior pituitaries of cycling, noncycling and early pregnant heifers. AB - In domestic animals limited data are available concerning levels of pituitary luteinizing hormone-releasing hormone (LHRH) receptors during various physiological states. The objectives of this study were to quantify anterior pituitary gonadotropin and LHRH receptor concentrations in cycling, noncycling and early pregnant beef heifers. To accomplish these objectives, five heifers each were slaughtered, after synchronization with prostaglandin F2 alpha (PGF2 alpha), on d 0 (estrus), 7 and 14 of the estrous cycle and d 40 of pregnancy. Four heifers determined to be noncycling were also slaughtered. Pituitaries were collected and analyzed for LHRH receptor and gonadotropin concentrations. Pituitary luteinizing hormone (LH) concentrations were low on d 0 (1.4 +/- .2 micrograms/mg pituitary, mean +/- SE) and remained low on d 7 (1.4 +/- .1 micrograms/mg pituitary) before increasing (P less than .01) on d 14 (2.6 +/- .5 micrograms/mg pituitary). Luteinizing hormone concentrations, compared with d 0, were also elevated (P less than .01) in noncycling (NC; 2.6 +/- .2 micrograms/mg pituitary) animals and in 40-d pregnant (PG; 2.5 +/- .2 micrograms/mg pituitary) heifers. Pituitary follicle stimulating hormone (FSH) concentrations, though similar (P greater than .05) for all groups, paralleled changes in LH concentration. Pituitary LHRH receptor binding affinity did not differ (P greater than .05) among groups, with an overall Kd = .64 +/- .02 X 10(-9) M. Luteinizing hormone-releasing hormone receptor concentrations were highest on d 0 (1.09 +/- .12 fmol/mg pituitary) and fell (P less than .01) to low levels on d 7 (.75 +/- .11 fmol/mg pituitary).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985530 TI - Effects of monensin, pyromellitic diimide and 2-bromoethanesulfonic acid on rumen fermentation in vitro. AB - The effects of monensin, 2-bromoethanesulfonic acid (2-BES) and pyromellitic diimide (diimide) on gas and volatile fatty acid (VFA) production by the rumen microbiota were compared in mixed culture. Oat hay, a hay-concentrate mixture (48% hay, 43% concentrate) and a soluble carbohydrate mixture were used as substrates for microbial growth. The highest concentrations of diimide (10 ppm) and 2-BES (30 microM) decreased methanogenesis by 97 and 76%, respectively, while H2 accumulation was increased 30- and 20-fold, respectively. The effect of monensin on methanogenesis was less dramatic as 10 ppm decreased CH4 accumulation 16% and H2 did not accumulate. Diimide and 2-BES decreased the acetate:propionate ratio with the hay (P less than .05) and soluble carbohydrate mixture (P less than .025). The numbers of saccharolytic, cellulolytic and methanogenic bacteria from sheep fed a diet with diimide (60 ppm) did not differ significantly from sheep fed a control diet. Semicontinuous culture incubations indicated that the mixed rumen microbial population could adapt and degrade diimide after 24 h of incubation. PMID- 2985531 TI - Effects of acetone in heparin on the multiple inert gas elimination technique. AB - We have detected acetone in several brands of heparin. If uncorrected, this leads to errors in measuring acetone in blood collected in heparinized syringes, as in the multiple inert gas elimination technique for measuring ventilation-perfusion ratio (VA/Q) distributions. Error for acetone retention [R = arterial partial pressure-to-mixed venous partial pressure (P-V) ratio] is usually small, because R is normally near 1.0, and the error is similar in arterial and mixed venous samples. However, acetone excretion [E = mixed expired partial pressure (P-E)-to P-V ratio] will appear erroneously low, because P-E is accurately measured in dry syringes, but P-V is overestimated. A physical model of a homogeneous alveolar lung at room temperature and without dead space shows: the magnitude of acetone E error depends upon the ratio of blood sample to heparinized saline volumes and acetone partial pressures, without correction, acetone E can be less than that of less soluble gases like ether, a situation incompatible with conventional gas exchange theory, and acetone R and E can be correctly calculated using the principle of mass balance if the acetone partial pressure in heparinized saline is known. Published data from multiple inert gas elimination experiments with acetone-free heparin, in our labs and others, are within the limits of experimental error. Thus the hypothesis that acetone E is anomalously low because of physiological mechanisms involving dead space tissue capacitance for acetone remains to be tested. PMID- 2985532 TI - Liquid chromatographic determination of vitamin D in milk and infant formula. AB - Vitamin D2 or vitamin D3 is determined by liquid chromatography (LC) in milk and infant formula. Vitamin D is extracted from the saponified sample, passed through an amino-cyano LC cleanup column to remove major interferences, and quantitated using normal phase LC. Within-day precision is 4.5% relative standard deviation (RSD); the overall method RSD (reflecting technician-to-technician, day-to-day, and within-day variability) is 7.7%. Overspike recoveries averaged 97% for milk, 98% for milk-based infant formula, and 93% for soy-based infant formula. The performance of the method is compared with that of the official AOAC vitamin D method (rat bioassay). The method is applicable to the determination of vitamin D in milk and in the major milk- and soy-based infant formulas available in the United States. The method can quantitate (but not distinguish) either vitamin D2 or vitamin D3. The method is applicable to milk and infant formula samples containing between 100 and 1500 IU vitamin D/L. Sample throughput is between 4 and 8 replicates per day. PMID- 2985533 TI - Liquid chromatographic determination of vitamins D2 and D3 in fortified milk and infant formulas. AB - A liquid chromatographic (LC) method was developed for determining vitamins D2 and D3 in fortified milk and infant formulas. The lipid-soluble components were extracted from the aqueous phase by homogenizing in isopropanol-methylene chloride with magnesium sulfate added to remove water. The vitamins were fractionated from the lipid material by using gel permeation chromatography (GPC) followed by further cleanup of the combined GPC fractions on a muBondapak/NH2 column. Four muStyragel (100 A) columns connected in series were used for GPC fractionation of sample extracts in methylene chloride. Injection and collection were repeated 3 times to collect enough vitamin D for quantitation. The muBondapak/NH2 column, using a mobile phase of methylene chloride-isooctane isopropanol (600 + 400 + 1), resolved vitamin D from other UV-absorbing compounds and soy sterols in infant formula and from cholesterol in milk. Vitamins D2 and D3 coeluted as one peak, with the resolution and vitamin level sufficient for visual monitoring (280 nm/0.02 absorbance unit full scale) in a collection time of 22-26 min. A Zorbax ODS (6 micron) column and a methylene chloride acetonitrile-methanol (300 + 700 + 2) mobile phase were used for LC quantitation; vitamins D2 and D3 were baseline resolved in about 11 min. The infant formula samples included ready-to-use and concentrated liquids prepared in nonfat milk base or soy base fortified with vitamins D2 or D3 at 400 IU/qt or L (10 micrograms). The mean percent recovery of added vitamin D3 (400-500 IU/qt) from infant formula (n = 7) was 89.6 +/- 6.7 (coefficient of variation (CV) 7.5%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985534 TI - Atomic absorption spectrophotometric determination of tin in canned foods, using nitric acid-hydrochloric acid digestion and nitrous oxide-acetylene flame: collaborative study. AB - Twenty-six collaborators participated in a study to evaluate an atomic absorption spectrophotometric (AAS) method for the determination of tin in canned foods. The 5 foods evaluated were meat, pineapple juice, tomato paste, evaporated milk, and green beans, each spiked at 2 levels. The concentration range of tin in the samples was 10-450 micrograms/g, and each level was sent as a blind duplicate. Statistical treatment of results revealed no laboratory outliers and 6 individual or replicate-total outliers, accounting for 3.3% of the data. Repeatability (within-laboratory) coefficient of variation (CVo) ranged from 2.2 to 48%, depending on the tin level and food evaluated. For samples containing greater than or equal to 80 micrograms/g of tin, repeatability CV averaged 5.6% including outliers and 3.7% after their rejection. Overall among-laboratories coefficient of variation (CVx) varied from 3.3 to 58%; at levels greater than or equal to 80 micrograms/g, it averaged 7.3% with outliers and 5.3% after their rejection. Recovery of tin, based on spiking levels, ranged from 100.0 to 112.8% and averaged 105.4%. Detection limit range is 2-10 micrograms/g, and lower quantitation limit is 40 micrograms/g. This method has been adopted official first action. PMID- 2985535 TI - Physical and genetic map of a Rhizobium meliloti nodulation gene region and nucleotide sequence of nodC. AB - Infection of alfalfa by the soil bacterium Rhizobium meliloti proceeds by deformation of root hairs and bacterial invasion of host tissue by way of an infection thread. We studied an 8.7-kilobase (kb) segment of the R. meliloti megaplasmid, which contains genes required for infection. Site-directed Tn5 mutagenesis was used to examine this fragment for nodulation genes. A total of 81 R. meliloti strains with mapped Tn5 insertions in the 8.7-kb fragment were evaluated for nodulation phenotype on alfalfa plants; 39 of the insertions defined a 3.5-kb segment containing nodulation functions. Of these 39 mutants, 37 were completely nodulation deficient (Nod-), and 2 at the extreme nif-distal end were leaky Nod-. Complementation analysis was performed by inoculating plants with strains carrying a genomic Tn5 at one location and a plasmid-borne Tn5 at another location in the 3.5-kb nodulation segment. Mutations near the right border of the fragment behaved as two distinct complementation groups. The segment in which these mutations are located was analyzed by DNA sequencing. Several open reading frames were found in this region, but the one most likely to function is 1,206 bases long, reading from left to right (nif distal to proximal) and spanning both mutation groups. The genetic behavior of this segment may be due either to the gene product having two functional domains or to a recombinational hot spot between the apparent complementation groups. PMID- 2985536 TI - Recombination in recA cells between direct repeats of insertion element IS1. AB - The IS1 sequences that flank the Tn9 chloramphenicol acetyltransferase gene as direct repeats recombine after transformation into an Escherichia coli recA strain. The recombination requires the lambda pL promoter on the plasmid. A plasmid that contains mutant IS1 elements does not recombine. These results indicate that this recombination requires an IS1-specific gene product. The recombinational activity of IS1 may resolve transient cointegrates formed during the transposition of IS1. I discuss a possible role for the lambda pL promoter. PMID- 2985537 TI - Repeated sequences similar to insertion elements clustered around the nif region of the Rhizobium japonicum genome. AB - Two different repeated sequences (RSs) were discovered in the Rhizobium japonicum genome: RSRj alpha is 1126 base pairs long and is repeated 12 times; RSRj beta is approximately 950 base pairs long and is repeated at least 6 times. Their arrangement in root nodule bacteroid DNA is the same as in DNA from bacteria grown in culture. Deletion analysis showed that many copies of alpha and beta are clustered around the nitrogenase genes nifDK and nifH, or, in general, they are found within a genomic region harboring genes that are nonessential for growth. One copy each of alpha and beta are located upstream of nifDK and are adjacent to each other. Neither of them, however, is involved in the expression of nifDK. Nucleotide sequence analysis of three copies of RS alpha revealed many characteristics of procaryotic insertion sequence elements: potential inverted repeats at their ends, potential target site duplication, and large open reading frames. Despite this, their genomic positions appear to be stable. One possible function of these RSs is in deletion formation probably via recombination between them. PMID- 2985538 TI - Physical analysis of deletion mutations in the ilvGEDA operon of Escherichia coli K-12. AB - DNA-DNA hybridization of cloned segments of the Escherichia coli K-12 ilvGEDA operon to genomic blots was used to determine the physical dimensions of a series of deletion mutations of the ilvGEDA operon. The smallest mutation resulted from the deletion of approximately 200 base pairs from within ilvD, whereas the largest mutation resulted from the deletion of 17 kilobases including the rep gene. The structure of three of these mutants indicates that formation of the deletions was mediated by Tn5 (or Tn5-131) that is retained in the chromosome. This is the first observation of this type of Tn5-mediated event. Our analysis of the total acetohydroxy acid synthase activity of strains containing deletions of ilvG indicates that the truncated ilvG polypeptide of wild-type E. coli K-12 lacks enzyme activity. The small 200-base-pair deletion of ilvD confirms the presence of a strong polar site 5' to ilvA. The detailed structure of these deletions should prove useful for the investigation of other genes in this region. This genomic analysis demonstrates that the ilv restriction site map that was established previously by the analysis of recombinant bacteriophage and plasmids is identical to that on the genome. PMID- 2985539 TI - Isolation of a novel transposon which carries the Escherichia coli enterotoxin STII gene. AB - The Escherichia coli heat-stable enterotoxin STII gene in P307 is flanked by inverted repeat sequences, suggesting that the STII gene is part of a transposon. To study the transposability, a DNA fragment containing the putative STII transposon has been cloned. Results of transposition assays indicated that the STII gene can transpose from one plasmid to another. The size of the transposon has been determined to be approximately 9 kilobases. The structure and the location of the STII gene in clinical isolates of Escherichia coli have been investigated by restriction enzyme analyses. The structural genes of STII from different clinical isolates appear to be uniform in size, but the flanking sequences are heterogeneous. This result suggests that the STII genes in different isolates are not on the same transposon as observed in P307. PMID- 2985540 TI - Evidence that the clindamycin-erythromycin resistance gene of Bacteroides plasmid pBF4 is on a transposable element. AB - We constructed a shuttle vector, pE5-2, which can replicate in both Bacteroides spp. and Escherichia coli. pE5-2 contains a cryptic Bacteroides plasmid (pB8-51), a 3.8-kilobase (kb) EcoRI-D fragment from the 41-kb Bacteroides fragilis plasmid pBF4, and RSF1010, an IncQ E. coli plasmid. pE5-2 was mobilized by R751, an IncP E. coli plasmid, between E. coli strains with a frequency of 5 X 10(-2) to 3.8 X 10(-1) transconjugants per recipient. R751 also mobilized pE5-2 from E. coli donors to Bacteroides uniformis 0061RT and Bacteroides thetaiotaomicron 5482 with a frequency of 0.9 X 10(-6) to 2.5 X 10(-6). The Bacteroides transconjugants contained only pE5-2 and were resistant to clindamycin and erythromycin. Thus, the gene for clindamycin and erythromycin resistance must be located within the Eco RI-D fragment of BF4. A second recombinant plasmid, pSS-2, which contained 33 kb of pBF4 (including the EcoRI-D fragment and contiguous regions) could also be mobilized by R751 between E. coli strains. In some transconjugants, a 5.5-kb (+/- 0.3 kb) segment of the pBF4 portion of pSS2 was inserted into one of several sites on R751. In some other transconjugants this same 5.5-kb segment was integrated into the E. coli chromosome. This segment could transfer a second time onto R751. Transfer was RecA independent. The transferred segment included the entire EcoRI-D fragment, and thus the clindamycin-erythromycin resistance determinant, from pBF4. PMID- 2985542 TI - Comparison of 10 IncP plasmids: homology in the regions involved in plasmid replication. AB - We have examined the DNA homology in the replication regions of 10 IncP plasmids independently isolated from several different countries. Two regions of RK2, the best-studied plasmid of this group, are required for vegetative DNA replication: the origin of replication (oriV) and the trfA region, which codes for a gene product necessary for replication. Six of nine IncP plasmids studied were identical to RK2 in the oriV and trfA regions as shown by Southern hybridization. Three P plasmids, R751, R772, and R906, showed weaker homology with the RK2 trfA, region and hybridized to different-sized HaeII fragments than the other six plasmids. R751, R772, and R906 hybridized to the region of the RK2 replication origin which expresses P incompatibility but differed markedly from RK2 and the other six plasmids in the GC-rich region of the origin required for replication. These data indicate that the P-group plasmids can be divided into two subgroups: IncP alpha, which includes the RK2-like plasmids, and IncP beta which includes the R751-like plasmids. PMID- 2985541 TI - Nucleotide sequence of ermA, a macrolide-lincosamide-streptogramin B determinant in Staphylococcus aureus. AB - The complete nucleotide sequence of ermA, the prototype macrolide-lincosamide streptogramin B resistance gene from Staphylococcus aureus, has been determined. The sequence predicts a 243-amino-acid protein that is homologous to those specified by ermC, ermAM, and ermD, resistance determinants from Staphylococcus aureus, Streptococcus sanguis, and Bacillus licheniformis, respectively. The ermA transcript, identified by Northern analysis and S1 mapping, contains a 5' leader sequence of 211 bases which has the potential to encode two short peptides of 15 and 19 amino acids; the second, longer peptide has 13 amino acids in common with the putative regulatory leader peptide of ermC. The coding sequence for this peptide is deleted in several mutants in which macrolide-lincosamide streptogramin B resistance is constitutively expressed. Potential secondary structures available to the leader sequence of the wild-type (inducible) transcript and to constitutive deletion, insertion, and point mutations provide additional support for the translational attenuation model for induction of macrolide-lincosamide-streptogramin B resistance. PMID- 2985543 TI - Isolation of a Bacillus stearothermophilus mutant exhibiting increased thermostability in its restriction endonuclease. AB - A procedure was developed for the selection of spontaneous mutants of Bacillus stearothermophilus NUB31 that are more efficient than the wild type in the restriction of phage at elevated temperatures. Inactivation studies revealed that two mutants contained a more thermostable restriction enzyme and one mutant contained three times more enzyme than the wild type. The restriction endonucleases from the wild type and one of the mutants were purified to apparent homogeneity. The mutant enzyme was more thermostable than the wild-type enzyme. The subunit molecular weight, amino acid composition, N-terminal and C-terminal amino acid residues, tryptic peptide map, and catalytic properties of the two enzymes were determined. The two enzymes have similar catalytic properties, but the molecular size of the mutant enzyme is approximately 6 to 7 kilodaltons larger than that of the wild-type enzyme. The mutant enzyme contains 54 additional amino acid residues, of which 26 to 28 are aspartate/asparagine, 8 to 15 are glutamate/glutamine, and 8 to 9 are tyrosine residues. The two enzymes contained similar amounts of the other amino acids, identical N-terminal residues, and different C-terminal residues. Tryptic peptide analyses revealed a high degree of homology between the two enzymes. The increased thermostability observed in the mutant enzyme appears to have been achieved by a mutation that resulted in the addition of amino acid residues to the wild-type enzyme. A number of mechanisms are discussed that could account for the observed difference between the mutant and wild-type enzymes. PMID- 2985544 TI - Isolation and characterization of Tn5 insertion mutants of Erwinia chrysanthemi that are deficient in polygalacturonate catabolic enzymes oligogalacturonate lyase and 3-deoxy-D-glycero-2,5-hexodiulosonate dehydrogenase. AB - Mutants of Erwinia chrysanthemi EC16 deficient in the polygalacturonate catabolic enzymes oligogalacturonate lyase (Ogl-) and 3-deoxy-D-glycero-2,5-hexodiulosonate (ketodeoxyuronate) dehydrogenase (KduD-) were obtained by Tn5 mutagenesis using the R plasmid pJB4JI. Ogl- Exu+ (Exu+, D-galacturonate utilization) and KduD- Exu strains macerated potato tuber tissue and utilized glucose, glycerol, and gluconate, but they did not utilize polygalacturonate, unsaturated digalacturonate, or saturated digalacturonate. Genetic and physical evidence indicated that the Ogl- mutants and a KduD- recombinant contained a single copy of Tn5 and that Tn5 (Kmr) was linked to the mutant phenotypes. In the Ogl+ parents, basal levels of oligogalacturonate lyase were present in glycerol-grown cells and induced levels were present with saturated or unsaturated digalacturonate, while oligogalacturonate lyase was undetectable under similar conditions in Ogl- strains. Pectate lyase, polygalacturonase, and ketodeoxyuronate dehydrogenase were induced in an Ogl- strain by 3-deoxy-D glycero-2,5-hexodiulosonate and by the enzymatic products of unsaturated digalacturonate but not by the digalacturonates. The KduD- strains lacked the dehydrogenase activity but in the presence of the digalacturonates produced higher levels of pectate lyase, polygalacturonase, and oligogalacturonate lyase than the KduD+ parents did. In the KduD- strains, pectate lyase and oligogalacturonate lyase were induced by unsaturated digalacturonate in a "gratuitous" manner, suggesting an intracellular accumulation of the inducer(s). We conclude that an intermediate(s) of the ketodeoxyuronate pathway induces pectate lyase, polygalacturonase, oligogalacturonate lyase, and ketodeoxyuronate dehydrogenase in E. chrysanthemi. PMID- 2985545 TI - Evidence for a common siderophore transport system but different siderophore receptors in Neurospora crassa. AB - Uptake and competition experiments were performed with Neurospora crassa and Penicillium parvum by using 14C-labeled coprogen and 55Fe-labeled ferrichrome type siderophores. Several siderophores of the ferrichrome family, such as ferrichrome, ferricrocin, ferrichrysin, and tetraglycyl-ferrichrome as well as the semisynthetic ferricrocin derivatives O-(phenyl-carbamoyl)-ferricrocin and O (sulfanilyl-carbamoyl)-ferricrocin were taken up by N. crassa. The ferrichrome type siderophores used vary in the structure of the peptide backbone but possess a common lambda-cis configuration about the iron center and three identical ornithyl-delta-N-acetyl groups as surrounding residues. This suggests that these ferrichrome-type siderophores are recognized by a common ferrichrome receptor. We also concluded that the ferrichrome receptor is lambda-cis specific from the inability to take up the synthetic enantiomers, enantio-ferrichrome and enantio ferricrocin, possessing a delta-cis configuration about the iron center. On the other hand, we found that coprogen, possessing a delta-absolute configuration and two trans-anhydromevalonic acid residues around the metal center, was also taken up by N. crassa and was competitively inhibited by the ferrichrome-type siderophores. We therefore propose the existence of a common siderophore transport system but the presence of different siderophore receptors in N. crassa. In addition, ferrirubin, which is very slowly transported by N. crassa, inhibited both coprogen and ferrichrome-type siderophore transport. Contrary to the findings with N. crassa, transport experiments with P. parvum revealed the presence of a ferrichrome receptor but the absence of a coprogen receptor; coprogen was neither transported nor did it inhibit the ferrichrome transport. PMID- 2985546 TI - Identification of three complementation units in the gerA spore germination locus of Bacillus subtilis. AB - The gerA locus, mutations in which affect the germination response of spores to L alanine and related amino acids, is contained within a 6-kilobase region of DNA cloned in phage and plasmid vectors. Fragments from this region, subcloned in the shuttle vector pHV33, were introduced into Bacillus subtilis, and their ability to complement chromosomal gerA mutations in a recE4 background was examined. Although the plasmids were somewhat unstable, it was possible to score complementation within spore-containing colonies on nutrient agar by their ability to reduce 2,3,5-triphenyltetrazolium chloride in an overlay. These studies have assigned the 10 gerA mutations tested to three complementation groups. An analysis of Tn1000 insertions into the cloned DNA of two relatively stable plasmids that together encompass the entire gerA region has identified more precisely the location and extent of the complementation units; recombination studies and in vitro mutagenesis were used to further delineate the extents of two of the units. The evidence suggests that the three complementation units are adjacent and that they are probably capable of separate transcription. PMID- 2985547 TI - Phasmid vectors for identification of genes by complementation of Escherichia coli mutants. AB - A bacteriophage lambda cloning vector was designed to facilitate the isolation of genes from procaryotic organisms by complementation of Escherichia coli mutants. This vector, lambda SE4, was constructed by attaching a very-low-copy-number replication system (from the plasmid NR1) and a spectinomycin resistance gene to the left arm of lambda 1059 (Karn et al., Proc. Natl. Acad. Sci. U.S.A. 77:5172 5176, 1980). This phasmid cloning vector is capable of growing lytically as a phage in a nonimmune host or lysogenically as a phasmid in an immune host. This phasmid utilizes the Spi- selection for insertions of DNA into the vector and has the ability to accept 2- to 19-kilobase Sau3A1, BamHI, BglII, BclI, or XhoII fragments; recombinants lysogenize immune hosts as single-copy-number selectable plasmids at 100% frequency. An E. coli library was constructed by using the initial vector lambda SE4, and clones of a number of representative genes were identified. A typical clone, lambda ant+, was shown to be readily mutagenized by a mini-Tn10 transposon. A general method for transferring cloned DNA segments onto bacteriophage lambda was developed. The method involves the use of in vivo recombination with a selection and was used to construct two derivatives of lambda SE4. Possible uses of these vectors and of the method for transferring cloned DNA onto phage lambda are discussed. PMID- 2985548 TI - Respiration-driven Na+ pump and Na+ circulation in Vibrio parahaemolyticus. AB - Sodium circulation in Vibrio parahaemolyticus was investigated. We observed respiration-driven Na+ extrusion from cells by using a Na+ electrode. The Na+ extrusion was insensitive to a proton conductor, carbonyl cyanide m chlorophenylhydrazone, and sensitive to a respiratory inhibitor, CN-. These results support the idea of the existence of a respiratory Na+ pump in V. parahaemolyticus. The respiration-driven Na+ extrusion was observed only under alkaline conditions. PMID- 2985549 TI - Allosteric regulation of glycerol kinase by enzyme IIIglc of the phosphotransferase system in Escherichia coli and Salmonella typhimurium. AB - The mechanism by which enzyme IIIglc of the bacterial phosphotransferase system regulates the activity of crystalline glycerol kinase from Escherichia coli has been studied, and the inhibitory effects have been compared with those produced by fructose-1,6-diphosphate. It was shown that the free, but not the phosphorylated, form of enzyme IIIglc inhibits the kinase. Mutants of Salmonella typhimurium were isolated which were resistant to inhibition by either enzyme IIIglc (glpKr mutants) or fructose-1,6-diphosphate (glpKi mutants), and each mutant type was shown to retain full sensitivity to inhibition by the other regulatory agent. Other mutants were fully or partially resistant to regulation by both agents. The two regulatory sites on the kinase are evidently distinct but must overlap or interact functionally. Kinetic analyses have revealed several mechanistic features of the regulatory interactions. (i) Inhibition by both allosteric regulatory agents is strongly pH dependent, with maximal inhibition occurring at ca. pH 6.5 under the assay conditions employed. (ii) Binding of enzyme IIIglc to glycerol kinase is also pH dependent, the Ki being near 4 microM at pH 6.0 but near 10 microM at pH 7.0. (iii) Whereas fructose-1,6-diphosphate inhibition apparently requires that the enzyme exist in a tetrameric state, both the dimer and the tetramer appear to be fully sensitive to enzyme IIIglc inhibition. (iv) Inhibition by enzyme IIIglc (like that by fructose-1,6 diphosphate) is noncompetitive with respect to both substrates. (v) The inhibitory responses of glycerol kinase to fructose-1, 6-diphosphate and enzyme IIIglc show features characteristic of positive cooperativity at low inhibitor concentration. (vi) Neither agent inhibits completely at high inhibitor concentration. (vii) Apparent negative cooperativity with respect to ATP binding is observed with purified E. coli glycerol kinase, with glycerol kinase in crude extracts of wild-type S. typhimurium cells, and with glpKr and glpKi mutant forms of glycerol kinase from S. typhimurium. These results serve to characterize the regulatory interactions which control the activity of glycerol kinase by fructose 1,6-diphosphate and by enzyme IIIglc of the phosphotransferase system. PMID- 2985550 TI - Preferential inhibition of plasmid replication in vivo by altered DNA gyrase activity in Escherichia coli. AB - The thermosensitive growth phenotype exerted by runaway-mutant plasmids was suppressed by sublethal doses of the DNA gyrase inhibitors novobiocin or nalidixic acid, although the latter drug was less efficient. A novobiocin resistant gyrB mutant Escherichia coli strain prevented expression of the runaway phenotype at 37 to 42 degrees C in the absence of any drug. PMID- 2985551 TI - A comparison of energy metabolism and the effects of beta-adrenergic stimulation in the tongue and attached gingiva of the dog. AB - Glucose-6-phosphate (G6P), ATP, phosphorylcreatine (CP) and lactate were assayed in the tongue attached gingiva of dogs. The G6P content was found to be significantly higher, and both the ATP and CP contents even more significantly higher in the tongue than in the attached gingiva. Lactate content was not significantly different in the gingiva and in the tongue. The changes in the concentrations of G6P, ATP, CP and lactate due to the intravenous infusion of isoproterenol hydrochloride (1 micrograms/kg/min) were as follows. In the tongue, G6P exhibited a significant increase, but the decrease in ATP and CP contents as well as the increase in lactate content were not significant. It is possible that these effects were the result of the activation of beta-adrenergic receptors by isoproterenol. In the attached gingiva, there was no significant change in the G6P, ATP, or CP content. Lactate increased to a greater degree than the other three components, but similarly did not reach the significant level. PMID- 2985552 TI - Effect of 1-p-bromotetramisole on phosphatases in neonatal hamster bone and tooth germs at different pH. AB - This study was designed to compare the various phosphatases and pyrophosphatases in bone with those in developing teeth. Moreover the alkaline phosphatase inhibitor 1-p-bromotetramisole (1-pBTM) was assessed for its ability to discriminate between the several phosphatase activities. Enzyme activities were determined over the pH range 3.50 to 11.25 in homogenates of calcifying hamster tibiae, and whole tooth germs using p-nitrophenyl-phosphate (pNPP) and pyrophosphate (PPi) as substrates. Characterization of alkaline phosphatase inhibition with 1-pBTM showed that in tooth germs the pI50 values of 1-pBTM for whole tooth germs, ectodermal and mesenchymal phosphatases were identical (pI50 = 5.36), which was slightly but significantly lower than that for bone alkaline phosphatase (pI50 = 5.64). In bone, two pNPP-ase activities were found with optima at pH 5.5 and pH 10.3 respectively. The alkaline phosphatase activity was completely inhibited by 10(-4) M 1-pBTM. Inhibition of the acid phosphatase was incomplete. With PPi as substrate, three activities were found with optima at pH 4.8, 7 and 8.7 respectively. All these PPi-ase activities were strongly inhibited by 1-pBTM in developing teeth, only one phosphatase activity was found, which exhibited an alkaline pH (10.3) optimum with both substrates. This activity was inhibited by 1-pBTM with both substrates although the effect on pNPP-ase activity was more marked. From these results we conclude that in bone there are insufficient differences in the extent of inhibition by 1-pBTM to distinguish between the various phosphatase activities when more physiological substrates such as pyrophosphate (PPi) are used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985553 TI - Stimulation by insulin-like growth factors is required for cellular transformation by type beta transforming growth factor. AB - Medium conditioned by BRL-3A cells, a known source of insulin-like growth factor II (IGF-II), induced phenotypic transformation (anchorage-independent proliferation) of mouse BALB/c 3T3 fibroblasts but not rat NRK-49F fibroblasts, in the presence of 10% calf serum. A specific radioreceptor assay and a bioassay indicated that BRL-3A conditioned medium contained 0.5-1 ng/ml of type beta transforming growth factor (beta TGF). Purified IGF-II and beta TGF acting together reconstituted the transforming activity of BRL-3A conditioned medium on BALB/c 3T3 cells. Insulin was 5-10% as potent as IGF-II in supporting the transforming action of beta TGF on BALB/c 3T3 cells. NRK-49F cells were phenotypically transformed by beta TGF in the presence of EGF and 10% calf serum as the sole source of IGFs. However, transformation of NRK-49F cells under these conditions was inhibited by addition of purified IGF-binding protein. Addition of an excess of IGF-II prevented the inhibitory action of IGF-binding protein. The different sensitivity of the two cell lines to IGFs was correlated with lower levels of type I IGF receptor and higher levels of type II IGF receptor in NRK 49F cells as compared with BALB/c 3T3 cells. The results suggest that cellular stimulation by IGFs is a prerequisite for transformation of rodent fibroblasts by beta TGF. We propose that transformation of fibroblasts by beta TGF requires concomitant stimulation by the set of growth factors that support normal cell proliferation. PMID- 2985554 TI - Studies on nuclease digestion of chromatin phosphorylated in vivo. AB - We have previously shown that, by culturing cells in hypertonic media, histone 2A becomes hyperphosphorylated (Pantazis, P., West, M. H. P., and Bonner, W. M. (1984) Mol. Cell. Biol. 4, 1186-1188). In the present study we have probed the effect of this histone modification on the overall chromatin structure by micrococcal nuclease and DNase I digestion. Although no significant quantitative differences in the extent of hydrolysis were observed between control and hyperphosphorylated chromatin by micrococcal nuclease, DNase I digested hyperphosphorylated chromatin at a 3- to 4-fold higher rate than unmodified chromatin. PMID- 2985555 TI - Homologous desensitization of beta-adrenergic receptor coupled adenylate cyclase. Resensitization by polyethylene glycol treatment. AB - Brief (approximately 20-min) exposure of S49 lymphoma cells to beta-agonists such as isoproterenol leads to a homologous form of desensitization in which beta agonist but not prostaglandin E1-sensitive or NaF-sensitive adenylate cyclase is reduced. The desensitized receptors (R) appear to be sequestered away from the effector system (guanine nucleotide regulatory protein (Ns) and adenylate cyclase (C)). Membrane perturbants such as polyethylene glycol are known to reorient membrane proteins and lipids. Thus, we fused agonist-desensitized S49 lymphoma cells to each other, using polyethylene glycol as fusogen, in an attempt to functionally reunite the R, N, and C components which might have become sequestered in microdomains of the plasma membrane during desensitization. Such treatment completely restored isoproterenol-stimulated adenylate cyclase to normal and re-established the ability of R and N to functionally couple as assessed by the ability to form a high affinity, guanine nucleotide-sensitive state of the receptor. These results support the concept that agonist-promoted sequestration plays a functionally significant role in the homologous desensitization of the beta-adrenergic receptor. PMID- 2985556 TI - Evidence for distinct catabolic pathways for deoxy-GTP and GTP in purine nucleoside phosphorylase-deficient mouse T lymphoblasts. AB - The catabolism of deoxy-GTP and GTP was compared in purine-nucleoside phosphorylase-deficient mouse T lymphoblasts. It was found that guanine ribonucleotides and deoxyribonucleotides are degraded by distinct pathways in cells cultured under both physiological and induced catabolic conditions. In T lymphoblasts, cultured under physiological conditions, 50% of the GMP formed during GTP catabolism was dephosphorylated and 50% was deaminated, whereas in the presence of the catabolic inducer deoxyglucose 90% of the GMP formed was dephosphorylated and only 10% was deaminated. These results indicate that GTP catabolism in lymphoblasts proceeds by alternative pathways, either via GMP dephosphorylation or via GMP reductive deamination, and physiological conditions determine with pathway will be used. In contrast, deoxy-GTP catabolism proceeds exclusively via deoxy-GMP dephosphorylation under both physiological and induced catabolic conditions. The lack of deoxy-GMP deamination may contribute to the accumulation of cytotoxic levels of deoxyguanosine found in purine-nucleoside phosphorylase-deficient patients. PMID- 2985557 TI - The gene for rat atrial natriuretic factor. AB - Atrial natriuretic factor (ANF), a peptide hormone recently isolated from heart atria, appears to play an important role in the regulation of extracellular fluid volume and blood pressure. Indeed, natural and synthetic ANF rapidly and markedly stimulate natriuresis and diuresis and produce smooth muscle relaxation. Consistent with the hypothesis that ANF is a novel hormone, it was recently shown that ANF is present in circulation, and high affinity membrane receptors specific for ANF have been described in renal, vascular, and adrenal tissues. These important biological activities suggest that conditions like hypertension could be associated with defective ANF gene expression. We and others have shown by cDNA cloning that ANF is part of a larger precursor, pro-natriodilatin (PND). We now describe the isolation and structural analysis of the rat PND gene. Southern blot analysis of rat genomic DNA suggests the presence of a single PND gene per haploid genome. The PND coding sequences are interrupted by two short introns. A long alternating purine-pyrimidine tract (GT)9GATG(GT)27 is found 111 base pairs downstream of the polyadenylation site; such sequences could adopt Z-DNA configuration and they have been associated with sequences that appear very active in intergenic recombination. Comparison of the rat and human PND genomic sequences shows highest homology in 5'-flanking as well as in coding sequences. The rat PND gene will be a useful model to study the physiology and pathology of this important regulator of the cardiovascular system. PMID- 2985558 TI - Linear electric field effect and electron spin-echo studies of uteroferrin. Evidence for iron coordination by a nitrogen-containing ligand. AB - Uteroferrin and semimethemerythrin, proteins possessing spin-coupled binuclear iron centers, exhibit large linear electric field effects in their mixed-valence, EPR-active states. This indicates that the paramagnetic center of each protein is noncentrosymmetric and suggests that charge may be localized on one of the iron atoms. The magnetic field dependence of the linear electric field effects for both proteins demonstrates that the direction of most facile polarization of the binuclear iron centers is near the orientation giving rise to gmin. Electron spin echo studies of uteroferrin reveal that its magnetic electron interacts with at least one and possibly two classes of nitrogen nuclei. Furthermore, comparison of echo envelope spectra for uteroferrin with that of ferric bleomycin suggests that one of these nuclei is from a histidine ligand. PMID- 2985559 TI - Omeprazole, a specific inhibitor of gastric (H+-K+)-ATPase, is a H+-activated oxidizing agent of sulfhydryl groups. AB - Omeprazole (5-methoxy-2-[[(4-methoxy-3,5- dimethylpyridinyl)methyl]sulfinyl]-1H benzimidazole) appeared to inhibit gastric (H+-K+)-ATPase by oxidizing its essential sulfhydryl groups, since the gastric ATPase inactivated by the drug in vivo or in vitro recovered its K+-dependent ATP hydrolyzing activity upon incubation with mercaptoethanol. Biological reducing agents like cysteine or glutathione, however, were unable to reverse the inhibitory effect of omeprazole. Moreover, acidic environments enhanced the potency of omeprazole. For example, in vivo pretreatment of rats with carbachol, a secretagogue, enhanced the activity of omeprazole to inhibit gastric (H+-K+)-ATPase, while pretreatment with cimetidine, an antisecretory agent, reduced its potency. In vitro, lowering pH of incubation media from 7.4 to 5.0 improved the ability of omeprazole to inhibit hog gastric (H+-K+)-ATPase almost 60-fold. The inhibitory effect of the drug was accompanied by a dose-dependently decreased amount of free sulfhydryl groups in the isolated hog gastric membranes. The chemical reactivity of omeprazole with mercaptans is also consistent with the biological action of omeprazole. The drug, only under acidic conditions, reacted with a stoichiometric amount of ethyl mercaptan (or beta-mercaptoethanol) to produce regio-isomers of N-sulfenylated omeprazole sulfide (5-methoxy-2[[(4-methoxy-3,5- dimethyl-2 pyridinyl)methyl]thio]-1- or 3-(ethylthio)benzimidazole). The N-sulfenylated compound reacted at neutral pH with another stoichiometric amount of ethyl mercaptan to produce omeprazole sulfide quantitatively. The gastric polypeptides of 100 kilodaltons representing (H+-K+)-ATPase in the rat gastric mucosa or isolated hog gastric membranes were covalently labeled with [14C]omeprazole. The radioactive label bound to the ATPase, however, could not be displaced by mercaptoethanol under the identical conditions where the ATPase activity was fully restored. These observations suggest that the essential sulfhydryl groups which reacted with omeprazole did not form a stable covalent bond with the drug, but rather that they further reacted with adjacent sulfhydryl groups to form disulfides which could be reduced by mercaptoethanol. PMID- 2985560 TI - Characterization of the chicken aldolase B gene. AB - Vertebrates possess three isozyme forms of fructose diphosphate aldolase. We have isolated two overlapping chicken genomic clones which encode the liver-specific form of this enzyme; we have identified the 5' and 3' ends of this gene by a combination of primer extension analysis and S1 mapping; and we have determined the entire nucleotide sequence of this gene including 1400 base pairs (bp) of sequence from the regions flanking the 5' and 3' ends of the gene. The transcriptional unit for the aldolase B gene spans 8700 bp and contains eight intervening sequences, including a 4600-bp intron in the 5' non-coding region. On the basis of results from Southern genomic hybridizations, the aldolase B gene appears to be present only once per haploid genome. No differences were detected in the mRNA structure between RNA from three tissues expressing aldolase B (liver, kidney, and small intestine). Various features of the 5' flanking region are discussed, including a partial homology with the 5' noncoding region from rabbit aldolase A. PMID- 2985561 TI - Signal recognition particle-dependent insertion of coronavirus E1, an intracellular membrane glycoprotein. AB - The membrane insertion of the E1 protein of a coronavirus, mouse hepatitis virus A59, was studied in a wheat germ cell-free translation system. E1 is a transmembrane protein spanning the lipid bilayer several times. It is synthesized without a cleavable signal sequence, localized intracellularly, and not transported to the cell surface. It thus represents a model intracellular protein. We found that the synthesis of E1 is specifically and stably blocked by the addition of signal recognition particle to the wheat germ system. Subsequent addition of salt-extracted pancreatic microsomes resulted in the full release of this arrest as well as the completion and the correct membrane integration of E1. Such signal recognition particle-induced arrests failed to produce shorter peptides of a defined length. Addition of signal recognition particle to a synchronized translation at any time during the synthesis of about the first two thirds of E1 (150 amino acids) blocked further translation, suggesting that the most C-terminal of the three internal hydrophobic domains of E1 could function as its signal sequence. PMID- 2985562 TI - Increase in cytosolic calcium and phosphoinositide metabolism induced by angiotensin II and [Arg]vasopressin in vascular smooth muscle cells. AB - Effects of angiotensin II and [Arg]vasopressin on cytosolic free Ca2+ concentration ([Ca2+]i) and phosphoinositide metabolism were studied in cultured aortic smooth muscle cells obtained from Wistar-Kyoto rats and their spontaneously hypertensive substrain. [Ca2+]i was measured using the fluorescent Ca2+ indicator quin2. No clear differences in basal [Ca2+]i were detected between cells derived from the two strains. High concentrations of angiotensin II (greater than or equal to 10 nM) and [Arg]vasopressin (greater than or equal to 100 nM) elicited large and rapid increases in [Ca2+]i, followed by a rapid return to control values. Low concentrations of these peptides (less than or equal to 1.0 nM) elicited small and slow increases in [Ca2+]i that persisted for minutes. These responses were blocked by specific antagonists for each of these peptides. Only high concentrations of angiotensin II caused [Ca2+]i increases in "Ca2+ free" medium, which suggested that high concentrations of angiotensin II could release Ca2+ from intracellular pools. A high concentration of angiotensin II and [Arg]vasopressin elicited progressive accumulations of inositol phosphates. Only high concentrations of angiotensin II caused inositol phosphate accumulation in Ca2+-free medium. Maximal accumulation of inositol phosphate elicited by angiotensin II and [Arg]vasopressin was found to be additive. A desensitization to the effects of both peptides on Ca2+ mobilization occurred despite the continued accumulation of inositol phosphates. These observations indicated that angiotensin II and [Arg]vasopressin interacted with independent receptors, both of which are linked to phosphoinositide breakdown and Ca2+ mobilization. PMID- 2985563 TI - The inactivation of ubiquitin accounts for the inability to demonstrate ATP, ubiquitin-dependent proteolysis in liver extracts. AB - The low molecular weight polypeptide required for energy-dependent proteolysis, ubiquitin, is rapidly inactivated by 100,000 X g supernatants of rabbit liver extracts. Ubiquitin inactivation results from limited proteolysis by an endogenous contaminating lysosomal thiol protease having trypsin-like specificity. Evidence for this includes a pH optimum of 5.0 for the first order constant of ubiquitin inactivation and observation that inactivation is inhibited by EDTA, o-phenanthroline, iodoacetamide, p-chloromercuribenzoic acid, phenylmethylsulfonyl fluoride, N alpha-p-tosyl-L-lysine chloromethyl ketone, leupeptin, soybean trypsin inhibitor, and aprotinin. Metals stimulate but are not required for ubiquitin inactivation with the effect apparently mediated by a low molecular weight heat-labile component of crude extracts. When this heat-labile component is removed by gel exclusion chromatography a number of metals inhibit ubiquitin inactivation. In the presence of excess dithiothreitol, inhibition is relatively specific for Zn(II). Inhibition by Zn(II) is specifically overcome competitively by Cd(II) or by a concentration of ubiquitin in excess of Zn(II). The responsible cathepsin possesses a molecular mass of 35 kDa by gel exclusion chromatography and shows marked thermal lability at neutral pH but stability at acid pH. Proteolytic inactivation of ubiquitin results from limited cleavage of the carboxyl-terminal glycine dipeptide required for isopeptide bond formation and is supported by data on isoelectric point changes on subsequent digestion with carboxypeptidase B and by direct amino acid analysis. When the responsible cathepsin is inactivated, liver extracts display ATP,ubiquitin-dependent proteolysis that cannot be ascribed to contaminating erythrocytes. Thus the previous inability to demonstrate energy-dependent proteolysis in liver extracts is accounted for by the artifactual inactivation of ubiquitin. PMID- 2985564 TI - Bacterial expression and characterization of proteins derived from the chicken calmodulin cDNA and a calmodulin processed gene. AB - Both normal chicken calmodulin (CaM) and a CaM-like mutant protein have been expressed in bacteria, isolated and evaluated with respect to several physical and biological properties. The mutant CaM is derived from a CaM-like gene that lacks intervening sequences and probably evolved from a CaM-processed gene (Stein, J. P., Munjaal, R. P., Lagace, L., Lai, E. C., O'Malley, B. W., and Means, A. R. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 6485-6489). The mutant CaM protein contains 16 of the 19 amino acids encoded by the CaM-like gene. Normal chicken CaM produced in bacteria is identical to rat CaM by all criteria tested except that it is not trimethylated. The protein product of the CaM-like gene has been termed CaML and exhibits properties which are very similar to CaM despite the presence of 16 amino acid substitutions. CaML binds Ca2+ as evidenced by Ca2+-dependent binding to phenothiazine- and phenyl-Sepharose affinity resins and a Ca2+-dependent electrophoretic mobility shift which is similar to but distinct from CaM. CaML cross-reacts with a monospecific CaM antibody and has an immunodilution curve which is identical to bacterially synthesized CaM. Finally, CaML can maximally activate rat brain phosphodiesterase but with altered kinetic parameters as compared to CaM. These data suggest that the nucleotide substitutions in the putative CaM processed gene are not random but are selected to retain CaM-like functions in the encoded protein. Such a mechanism may exist for other processed genes. PMID- 2985565 TI - A single amino acid substitution in the enzyme 5-enolpyruvylshikimate-3-phosphate synthase confers resistance to the herbicide glyphosate. AB - The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19), encoded by the aroA locus, is a target site of glyphosate inhibition in bacteria. A glyphosate-resistant aroA allele has been cloned in Escherichia coli from a mutagenized strain of Salmonella typhimurium. Subcloning of this mutant aroA allele shows the gene to reside on a 1.3-kilobase segment of S. typhimurium DNA. Nucleotide sequence analysis of this mutant gene indicates a protein-coding region 427 amino acids in length. Comparison of the mutant and wild type aroA gene sequences reveals a single base pair change resulting in a Pro to Ser amino acid substitution at the 101st codon of the protein. A hybrid gene fusion between mutant and wild type aroA gene sequences was constructed. 5-Enolpyruvylshikimate 3-phosphate synthase was prepared from E. coli cells harboring this construct. The glyphosate-resistant phenotype is shown to be associated with the single amino acid substitution described above. PMID- 2985566 TI - Characterization of an Escherichia coli mdoB mutant strain unable to transfer sn 1-phosphoglycerol to membrane-derived oligosaccharides. AB - A procedure for the isolation of mutants affected in components containing glycerol derived from phospholipids yielded two mutant strains that contain membrane-derived oligosaccharides (MDO) devoid of glycerol (Rotering, H., Fiedler, W., Rollinger, W., and Braun, V. (1984) FEMS Microbiol. Lett. 22, 61 68). MDO are found in the periplasmic space of Escherichia coli and other Gram negative bacteria, and they may comprise up to 7% of the cells dry weight. The biosynthesis of MDO is osmoregulated (Kennedy, E. P. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 1092-1095) and linked to the metabolism of phospholipids (van Golde, L. M. G., Schulman, H., and Kennedy, E. P. (1973) Proc. Natl. Acad. Sci. U. S. A. 70, 1368-1372). This leads to substitution of MDO with sn-1 phosphoglycerol and phosphoethanolamine (Kennedy, E. P., Rumley, M. K., Schulman, P., and van Golde, L. M. G. (1976) J. Biol. Chem. 251, 4208-4213). MDO also contain succinate in O-ester linkage. We now report that one mutant strain lacks phosphoglycerol transferase I activity and thus is unable to transfer sn-1 phosphoglycerol residues from phosphatidylglycerol to MDO. The mdoB gene affected in this mutant has been located at 99.2 min on the E. coli chromosome. The ethanolamine content of MDO isolated from the mutant strain is elevated, whereas the number of succinate residues is not affected. The only phenotype of mdoB mutants we found is a dramatic reduction of the diglyceride content observed in dgk mdoB double mutants when the beta-glucoside arbutin is present in the growth medium. PMID- 2985567 TI - The mechanism of pantothenate transport by rat liver parenchymal cells in primary culture. AB - The mechanism of pantothenate transport across the plasma membrane was investigated with initial velocity studies of [14C]pantothenate uptake and efflux in rat liver parenchymal cells maintained in primary culture. At 116 mM sodium, double-reciprocal plots of the initial velocity of uptake versus [pantothenate] were linear from 0.3 to 36.5 microM pantothenate and gave an apparent Km,pant of 11 +/- 2 microM. The rate of pantothenate uptake at 0 [sodium] was about 14% of the rate at 116 mM sodium, and the reciprocal of the apparent Km,pant was a linear function of [sodium]. Vmax obtained by extrapolation to infinite [pantothenate] was independent of [sodium]. Ouabain, gramicidin D, cyanide, azide, and 2,4-dinitrophenol inhibited uptake, but preloading cells with pantothenate did not. Pantothenate derivatives or carboxylic acids were only weak inhibitors of uptake. Efflux was measured in cells preloaded with [14C]pantothenate. The apparent Km for efflux was 85 +/- 29 microM, and the rate of efflux was unaffected by addition of pantothenate, sodium, ouabain, gramicidin D, or 2,4-dinitrophenol to the external medium. These features are consistent with a mechanism for pantothenate transport in which sodium and pantothenate are cotransported in a 1:1 ratio on a carrier highly specific for pantothenate; sodium decreases the apparent Km for pantothenate, and a sodium-carrier complex forms only on the intracellular side of the membrane. PMID- 2985569 TI - Role of a Na+-dependent Cl-/HCO3- exchange in regulation of intracellular pH in fibroblasts. AB - We previously reported that, in a HCO3(-)-free medium, cytoplasmic pH (pHi) of hamster fibroblasts (CCL39) is primarily regulated by an amiloride-sensitive Na+/H+ antiport (L'Allemain, G., Paris, S., and Pouyssegur, J. (1984) J. Biol. Chem. 259, 5809-5815). Here we demonstrate the existence of an additional pHi regulating mechanism in CCL39 cells, namely a Na+-dependent HCO3-/Cl- exchange. Evidence for this system is based on 36Cl- influx studies and on pHi measurements in PS120, a CCL39-derived mutant lacking the Na+/H+ antiport activity. 36Cl- influx rate is a saturable function of external [Cl-] (apparent Km approximately equal to 7 mM), is competitively inhibited by external HCO3- (KI approximately equal to 3 mM), and by stilbene derivatives (KI approximately equal to 20 microM for 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid). Measurements of pHi recovery after an acute acid load indicate that PS120 cells possess an acid extruding mechanism dependent on external HCO3-, which is inhibited by stilbene derivatives and requires external Na+. Since 22Na+ influx is stimulated upon addition of HCO3- to acid-loaded cells and this effect is completely abolished by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, we conclude that Na+ is co transported with HCO3-, in exchange for intracellular Cl-. In a HCO3(-) containing medium, this pHi-regulating mechanism appears to have two essential physiological functions for the Na+/H+ antiport-deficient mutant: protection of the cells against excessive cytoplasmic acidification and establishment of a steady-state pHi permissive for growth, at neutral or slightly acidic pHo values (6.6-7.2). PMID- 2985568 TI - Demonstration of a Na+/H+ exchange activity in purified canine cardiac sarcolemmal vesicles. AB - Purified canine cardiac sarcolemmal membrane vesicles exhibit a sodium ion for proton exchange activity (Na+/H+ exchange). Na+/H+ exchange was demonstrated both by measuring rapid 22Na uptake into sarcolemmal vesicles in response to a transmembrane H+ gradient and by following H+ transport in response to a transmembrane Na+ gradient with use of the probe acridine orange. Maximal 22Na uptake into the sarcolemmal vesicles (with starting intravesicular pH = 6 and extravesicular pH = 8) was approximately 20 nmol/mg protein. The extravesicular Km of the Na+/H+ exchange activity for Na+ was determined to be between 2 and 4 mM (intravesicular pH = 5.9, extravesicular pH = 7.9), as assessed by measuring the concentration dependence of the 22Na uptake rate and the ability of extravesicular Na+ to collapse an imposed H+ gradient. All results suggested that Na+/H+ exchange was reversible and tightly coupled. The Na+/H+ exchange activity was assayed in membrane subfractions and found most concentrated in highly purified cardiac sarcolemmal vesicles and was absent from free and junctional sarcoplasmic reticulum vesicles. 22Na uptake into sarcolemmal vesicles mediated by Na+/H+ exchange was dependent on extravesicular pH, having an optimum around pH 9 (initial internal pH = 6). Although the Na+/H+ exchange activity was not inhibited by tetrodotoxin or digitoxin, it was inhibited by quinidine, quinacrine, amiloride, and several amiloride derivatives. The relative potencies of the various inhibitors tested were found to be: quinacrine greater than quinidine = ethylisopropylamiloride greater than methylisopropylamiloride greater than dimethylamiloride greater than amiloride. The Na+/H+ exchange activity identified in purified cardiac sarcolemmal vesicles appears to be qualitatively similar to Na+/H+ exchange activities recently described in intact cell systems. Isolated cardiac sarcolemmal vesicles should prove a useful model system for the study of Na+/H+ exchange regulation in myocardial tissue. PMID- 2985570 TI - Heterologous desensitization of adenylate cyclase with prostaglandin E1 alters sensitivity to inhibitory as well as stimulatory agonists. AB - Adenylate cyclase in cultured human fibroblasts is activated by prostaglandin E1 (PGE1) or beta-adrenergic agonists, e.g., isoproterenol, and inhibited by muscarinic agonists. Incubation with PGE1 reduced adenylate cyclase responsiveness to both PGE1 and isoproterenol; this so-called heterologous desensitization is believed to result from impaired function of the stimulatory guanyl nucleotide-binding protein of the cyclase complex. The effect of heterologous desensitization by PGE1 on inhibition of adenylate cyclase by the muscarinic agonist oxotremorine was examined. Muscarinic inhibition of basal and isoproterenol-stimulated cAMP accumulation was attenuated following exposure to PGE1; the concentration of oxotremorine required for half-maximal inhibition of cAMP accumulation was increased. In both intact cells and membrane preparations the number of binding sites for [3H]scopolamine, a muscarinic antagonist, was unaltered by desensitization. Following exposure to PGE1, receptor affinity for oxotremorine, assessed by competition with [3H] scopolamine, and the guanyl nucleotide sensitivity of agonist binding were reduced. The amount of inhibitory guanyl nucleotide-binding regulatory protein available for [32P]ADP-ribosylation by pertussis toxin was unaltered by desensitization. Thus, heterologous desensitization of adenylate cyclase with the stimulatory agonist PGE1 alters sensitivity to inhibitory as well as stimulatory ligands. PMID- 2985572 TI - Mechanism of strand passage by Escherichia coli topoisomerase I. The role of the required nick in catenation and knotting of duplex DNA. AB - We studied the interaction between topoisomerase I and a nicked DNA substrate to determine how the nick permits Escherichia coli topoisomerase I to catenate and knot duplex DNA rings. The presence of just a single nick in a 6600-base pair DNA increased the amount of DNA bound to topoisomerase I by 6-fold. The enzyme acts at the nick, as shown by linearization of nicked circles and covalent attachment of an enzyme molecule opposite the nick. DNA breaks are also introduced by the enzyme at sites not opposite to a nick, but three orders of magnitude less efficiently. The break induced by the enzyme is within several base pairs of the nick and on the complementary strand, but the exact site cut is dictated by DNA sequence requirements. Because these sequence requirements are identical to those for cutting of single-stranded DNA, we conclude that the enzyme stabilizes a denatured region at the nick. Breaks in single-stranded DNA occur 98% of the time when a C residue is four bases to the 5' side unless G is adjacent and 5' to the break. For a DNA circle nicked at a unique location, the efficiency of DNA breakage opposite the nick correlates with the rate of catenation. We present a unified model for the relaxation, catenation, and knotting reactions of topoisomerase I in which the enzyme induces a break in a single-stranded region, but bridges that break with covalent and noncovalent interactions and allows passage of one duplex or single-stranded DNA segment. PMID- 2985571 TI - Duplex DNA knots produced by Escherichia coli topoisomerase I. Structure and requirements for formation. AB - We investigated systematically the knotting of nicked circular duplex DNA by Escherichia coli topoisomerase I. Agarose gel electrophoresis of knots forms a ladder of DNA bands. Each rung is made up of a variety of knots with the same number of nodes, or segment crossings; knots in adjacent rungs differ by one node. We extended the technique of electron microscopy of recA protein-coated DNA to the visualization of the complex knots tied by topoisomerase I. The striking result is that the enzyme produces every knot theoretically possible. The requirement for excess enzyme to form complex knots suggests a role for topoisomerase I in contorting the DNA in addition to promoting strand passage. We conclude that nodes formed are equally likely to be positive or negative and that topoisomerase I can pass DNA strands through a transient enzyme-generated break without regard to orientation of the passing strand. The results are interpreted in terms of a formulation for the topological requirements for knotting. PMID- 2985573 TI - Epidermal growth factor receptor of A431 cells. Characterization of a monoclonal anti-receptor antibody noncompetitive agonist of epidermal growth factor action. AB - A monoclonal antibody to the epidermal growth factor (EGF) receptor of A431 cells, denoted 2D1-IgM, was generated after fusion of immunized BALB/c mouse spleen cells with SP2/0-Ag14 myeloma cells. Specific binding of 2D1-IgM to the A431 cell-surface receptor for EGF was demonstrated by indirect immunofluorescence, immunoprecipitation, and immunoblot analysis. Scatchard analysis of 125I-EGF binding to A431 cells demonstrated that 2D1-IgM treatment did not change the number of EGF receptors, but caused an increase in the affinity of EGF receptors from a population of low affinity to a uniform population of high affinity. Like EGF, 2D1-IgM induced phosphorylation of EGF receptors and EGF receptor clustering. As in the case of EGF, a biphasic growth response with stimulation of DNA synthesis at low and inhibition at high concentrations of 2D1-IgM was evident in A431 cells. The intrinsic "EGF-like" bioactivity of 2D1-IgM was enhanced by the presence of EGF. These results suggest that the binding of 2D1-IgM to the EGF receptor at a different site from that to which EGF binds can initiate an effective EGF-like biological response; and the EGF-like biological effects of 2D1-IgM may be mediated by a population of high affinity EGF receptors which may be involved in the control of cellular growth. PMID- 2985574 TI - Complete nucleotide sequence of a methylcholanthrene-inducible cytochrome P-450 (P-450d) gene in the rat. AB - The rat cytochrome P-450d gene which is inducibly expressed by the administration of 3-methylcholanthrene (MC) has been cloned and analyzed for the complete nucleotide sequence. The gene is 6.9 kilobases long and is separated into 7 exons by 6 introns. The insertion sites of the introns in this gene are well-conserved as compared with those of another MC-inducible cytochrome P-450c gene, but are completely different from those of a phenobarbital-inducible cytochrome P-450e gene. The overall homologies in the coding nucleotide and deduced amino acid sequences were 75% and 68% between the two MC-inducible cytochrome P-450 genes, respectively. The similarity of the gene organization between cytochrome P-450d and P-450c as well as their homology in the deduced amino acid and the nucleotide sequences suggests that these two genes of MC-inducible cytochromes P-450 constitute a different subfamily than those of the phenobarbital-inducible one in the cytochrome P-450 gene family. In contrast with the notable sequence homology in the coding region of the two MC-inducible cytochromes P-450, all the introns and the 5'- and 3'-flanking regions of the two genes showed virtually no sequence homology between them except for several short DNA segments that are located in the promoter region and the first intron. The nucleotide sequences and the locations of these conserved short DNA segments in the two genes suggest that they may affect the expression of the genes. Middle repetitive sequence reported as ID or identifier sequence were found in and in the vicinity of the cytochrome P-450d gene. PMID- 2985575 TI - Identification of ubiquitinated histones 2A and 2B in Physarum polycephalum. Disappearance of these proteins at metaphase and reappearance at anaphase. AB - Ubiquitinated histones uH2A.1, uH2A.Z, and uH2B have been identified in the basic nuclear proteins of the slime mold Physarum polycephalum by three methods: peptide mapping, cross-reaction with anti-ubiquitin antibody, and uH2A and uH2B isopeptidase cleavage. In microplasmodia, uH2A amounts to 7% of H2A and uH2B amounts to 6% of H2B. Detailed studies of mitosis in Physarum polycephalum macroplasmodia show that in early prophase, which last 15 min, the uH2As and uH2B are both strongly present, whereas minutes later in metaphase, which lasts 7 min, they disappear. When the nuclei enter anaphase, which lasts 3 min, both the uH2As and uH2B reappear. These precise studies suggest that cleavage of ubiquitin from the uH2As and uH2B is a very late, possibly final event in chromosome condensation to metaphase chromosomes and that ubiquitination is an early event in their decondensation. It is proposed that the uH2A and uH2B mark specific regions of the genome which have to be deubiquitinated prior to packaging into metaphase chromosomes; after metaphase these regions are the first to be decondensed and ubiquitinated. This modification, however, is not thought to be a general factor in chromosome condensation but labels a specific subcomponent of chromatin containing the expressed genes of a particular cell type or an important subset of these genes required by the cell to be available for activation, e.g. stress genes. PMID- 2985576 TI - Identification and characterization of calmodulin-binding proteins in islet secretion granules. AB - The binding of 125I-calmodulin to intact secretion granules and protein gel blots of secretion granules from pancreatic islet tissue was examined. Binding of 125I calmodulin to intact secretion granules was Ca2+-dependent and inhibited by the calmodulin inhibitors trifluoperazine and calmidazolium. Binding was inhibited by excess (200 nM) unlabeled calmodulin, but not by parvalbumin, a Ca2+-binding protein which has little sequence homology to calmodulin. In order to study the binding of calmodulin to specific secretion granule proteins, secretion granules were solubilized, and the solubilized proteins were resolved on sodium dodecyl sulfate-polyacrylamide gels, electrophoretically transferred to nitrocellulose, and incubated with 125I-calmodulin. Autoradiograms of the protein gel blots revealed the presence of three major calmodulin-binding proteins with approximate molecular weights of 73,000, 64,000, and 58,000. These proteins reversibly bound calmodulin in a calcium-dependent manner. Unlabeled calmodulin in the range of 0.1-1.0 nM competed with 125I-calmodulin for binding to these proteins, whereas troponin and parvalbumin were 100 and 1000-fold less effective, respectively. Trifluoperazine blocked binding to the granule proteins in a range of 10(-4) to 10(-5) M, and calmidazolium was effective between 10(-5) and 10(-6) M. Trypsin, at a concentration which did not lyse granules, markedly inhibited calmodulin binding to intact secretion granules. Protein blots from trypsin-treated granules showed that the three major calmodulin-binding proteins were absent. These results indicate that Ca2+-dependent calmodulin-binding proteins are present on the cytoplasmic surface of islet secretion granules and are consistent with the hypothesis that these proteins may play a role in secretion granule exocytosis. PMID- 2985577 TI - Complex formation between flavodoxin and cytochrome c. Cross-linking studies. AB - Complex formation between Azotobacter vinelandii flavodoxin and horse cytochrome c has been demonstrated through cross-linking studies with dimethyl suberimidate, dimethyl adipimidate, 1-ethyl-3-(3-di-methylaminopropyl)carbodiimide, and dimethyl-3,3'-dithiobispropionimidate. Essentially quantitative cross-linking of cytochrome c and flavodoxin was observed at low ionic strengths with the carbodiimide cross-linking reagent. An association constant of 4 X 10(4) M-1 was obtained between cytochrome c and flavodoxin at 88 mM ionic strength from analysis of the cross-linking studies. This value is similar to the association constant determined kinetically during the electron transfer reaction between cytochrome c and flavodoxin (Simondsen, R.P., Weber, P.C., Salemme, F.R., and Tollin, G. (1982) Biochemistry 21, 6366-6375), and suggests that the cross-linked complex may be similar to the precursor complex identified kinetically. A structural model for the flavodoxin-cytochrome c complex proposed by these workers is shown to be compatible with the present cross-linking results. PMID- 2985578 TI - Structure of an electron transfer complex. I. Covalent cross-linking of cytochrome c peroxidase and cytochrome c. AB - Cytochrome c peroxidase and cytochrome c form a noncovalent electron transfer complex in the course of the peroxidase-catalyzed reduction of H2O2. The two hemoproteins were cross-linked in 40% yield to a covalent 1:1 complex with the aid of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The covalent complex was found to be a valid model of the noncovalent electron transfer complex for the following reasons. The covalent complex had only 5% residual peroxidase activity toward exogeneous ferrocytochrome c indicating that the cross-linked cytochrome c covers the electron-accepting site of cytochrome c peroxidase. The residual peroxidase activity was almost independent of ionic strength indicating that the electron-accepting site is much less accessible even when ionic bonds between the two cross-linked hemoproteins are severed. The rate of reduction of heme c by ascorbate is 15 times slower in the covalent complex than in free cytochrome c and is independent of ionic strength. Although the covalent complex may not have been entirely pure with respect to the number and location of the cross-links, two major cross-links could be localized to within a few residues. One is from Lys 13 of cytochrome c to an acidic residue in positions 32, 33, 34, 35, or 37 of cytochrome c peroxidase, the other from Lys 86 of cytochrome c to a carboxyl group in the same cluster of acidic residues. The result stresses the importance of a peculiar stretch of acidic residues of cytochrome c peroxidase and of Lys 13 and 86 of cytochrome c. PMID- 2985579 TI - Structure of an electron transfer complex. II. Chemical modification of carboxyl groups of cytochrome c peroxidase in presence and absence of cytochrome c. AB - Cytochrome c peroxidase forms an electron transfer complex with cytochrome c. The complex is governed by ionic bonds between side chain amino groups of cytochrome c and carboxyl groups of peroxidase. To localize the binding site for cytochrome c on the peroxidase, we have used the method of differential chemical modification. By this method the chemical reactivity of carboxyl groups (toward carbodiimide/aminoethane sulfonate) was compared in free and in complexed peroxidase. When ferricytochrome c was bound to cytochrome c peroxidase, acidic residues 33, 34, 35, 37, 221, 224, and 1 to 3 carboxyls at the C terminus became less reactive by a factor of approximately 4, relative to the remaining 39 carboxylates of peroxidase. Of the less reactive residues those in the 30-40 region and the 221/224 pair are on opposite sides of the surface area which contains the heme propionates. We, therefore, propose that the binding site for cytochrome c on cytochrome c peroxidase spans the area where one heme edge comes close to the molecular surface. The results are in very good agreement with chemical cross-linking studies (Waldmeyer, B., and Bosshard, H.R. (1985) J. Biol. Chem. 260, 5184-5190); they also support a hypothetical model predicted on the basis of the known crystal structures of cytochrome c and peroxidase (Poulos, T.L., and Kraut, J. (1980) J. Biol. Chem. 255, 10322-10330). PMID- 2985580 TI - Identification of residues in the nucleotide binding site of the epidermal growth factor receptor/kinase. AB - We have purified the epidermal growth factor receptor/kinase from A431 membrane vesicles which had been affinity labeled with the ATP analog, 5'-p fluorosulfonylbenzoyl[8-14C]adenosine. The resulting purified, affinity labeled receptor/kinase preparation has been subjected to reduction and carboxymethylation followed by tryptic digestion. From this digest, we have isolated and sequenced the tryptic peptide containing the major site of labeling by the ATP analog. The sequence of this peptide is Ile-Pro-Val-Ala-Ile-X-Glu-Leu, where X corresponds to Lys 721 of the derived sequence of the EGF receptor/kinase. PMID- 2985581 TI - Induction of collagenase secretion in human fibroblast cultures by growth promoting factors. AB - In human foreskin fibroblast cultures, two proteins with Mr 60,000 and 55,000 were found to be induced about 3.5-fold by epidermal growth factor (EGF), platelet-derived growth factor, and beta-transforming growth factor. The induced proteins were identified as procollagenases by immunoprecipitation of induced medium with antibodies to purified human fibroblast collagenase. Collagenase enzyme activity in the medium from EGF-treated cultures was also induced at least 3-fold compared to control cultures. Induction of collagenase was dependent upon de novo protein and RNA synthesis and was observed in the medium 10 h after addition of EGF. Although these growth-promoting factors interact with separate membrane receptors, each induced the secretion of a common protein, suggesting that collagenase may be important in some aspect of mitogenesis, cell mobilization, and migration. PMID- 2985582 TI - The C-6 proton of tetrahydrobiopterin is acquired from water, not NADPH, during de novo biosynthesis. AB - Tetrahydrobiopterin, the cofactor for the aromatic amino acid hydroxylases, is synthesized in mammals from GTP via a pathway involving both dihydropterin and tetrahydropterin intermediates. In this work, we have investigated the mechanism of conversion of the product formed from GTP, 7,8-dihydroneopterin triphosphate, into the tetrahydropterin intermediates. Tetrahydrobiopterin can be oxidized under conditions which yield pterin or pterin 6-carboxylate without exchange of the C-6 and C-7 protons. Using these techniques, a gas chromatography/mass spectrometry method was developed to determine that in the biosynthesis of tetrahydrobiopterin de novo, in preparations of bovine adrenal medulla, the C-6 proton of tetrahydrobiopterin is derived from water and not from NADPH. In contrast, the C-6 proton of tetrahydrobiopterin produced from sepiapterin (6 lactoyl-7,8-dihydropterin) comes from NADPH. The results are consistent with evidence for the formation of the first tetrahydropterin intermediate by a tautomerization without any requirement for NADPH. PMID- 2985583 TI - Specific recognition of calmodulin from Dictyostelium discoideum by the ATP, ubiquitin-dependent degradative pathway. AB - Calmodulin purified from Dictyostelium discoideum is selectively degraded by rabbit reticulocyte extracts in the presence of ubiquitin and ATP. This protein forms a 1:1 covalent conjugate with ubiquitin. Analyses of the cyanogen bromide fragments of the protein conjugate indicate that lysine 115 on calmodulin is the ubiquitin conjugation site. Bovine brain calmodulin which contains a trimethyllysine residue at this position is not a substrate for conjugation with ubiquitin, and its degradation rate is not affected by ATP and ubiquitin. These results suggest that the trimethyllysine residue in mammalian calmodulin may function in protecting the protein from degradation by the ATP, ubiquitin dependent pathway. Since there are eight lysine residues in Dictyostelium calmodulin, the specific conjugation of ubiquitin to lysine 115 may provide a good model system to delineate the structural features required for the conjugation and to follow the degradative steps in the pathway. PMID- 2985584 TI - Phorbol ester inhibits phosphoinositide hydrolysis and calcium mobilization in cultured astrocytoma cells. AB - In cultured human 1321N1 astrocytoma cells, muscarinic receptor stimulation leads to phosphoinositide hydrolysis, formation of inositol phosphates, and mobilization of intracellular Ca2+. Treatment of these cells with 1 microM 4 beta phorbol 12 beta-myristate 13 alpha-acetate (PMA) completely blocks the carbachol stimulated formation of [3H]inositol mono-, bis-, and trisphosphate ( [3H]InsP, [3H]InsP2, and [3H]InsP3). The concentrations of PMA that give half-maximal and 100% inhibition of carbachol-induced [3H]InsP formation are 3 nM and 0.5 microM, respectively. Inactive phorbol esters (4 alpha-phorbol 12,13-didecanoate and 4 beta-phorbol), at 1 microM, do not inhibit carbachol-stimulated [3H]InsP formation. The KD of the muscarinic receptor for [3H]N-methyl scopolamine is unchanged by PMA treatment, while the IC50 for carbachol is modestly increased. PMA treatment also abolishes carbachol-induced 45Ca2+ efflux from 1321N1 cells. The concomitant loss of InsP3 formation and Ca2+ mobilization is strong evidence in support of a causal relationship between these two responses. In addition, our finding that PMA blocks hormone-stimulated phosphoinositide turnover suggests that there may be feedback regulation of phosphoinositide metabolism through the Ca2+- and phospholipid-dependent protein kinase. PMID- 2985585 TI - Role of cell membrane composition in receptor-mediated internalization of vesicular stomatitis virus in human HEp-2 cells. AB - The role of essential fatty acids in membrane functions related to receptor mediated endocytosis of vesicular stomatitis virus (VSV) was investigated using a human laryngeal carcinoma cell line (HEp-2) grown in chemically defined serum free medium (DM) to deplete their essential fatty acid contents. VSV replicated much less effectively in HEp-2 cells grown in DM as compared to serum containing complete medium (CM). Observed reduction in the rate of virus multiplication was, at least in part, due to reduced virus penetration which was monitored using VSV labeled with nitroxyl free radicals as electron spin probe. Surface proteins of VSV were labeled with maleimide spin-label, and succinimide spin-label. Ni2+ was used as a broadening agent to identify the spin-label signals from viruses inside the cell. HEp-2 cells and mouse leukemia cell line L1210 treated with 5 dimethylaminonaphthalene-1-sulfonyl (dansyl) cadaverine, an agent previously shown to inhibit the uptake of VSV in vitro, was used as a positive control in some experiments. VSV penetrated less effectively in both DM-grown cells and in CM-grown cells in the presence of dansylcadaverine. Similar results were obtained by monitoring the uptake of 125I-labeled VSV. When HEp-2 cells grown for several generations in DM were incubated with 10% fetal calf serum for 16 h, the cells supported virus replication to a similar extent as the cells grown in CM. In contrast, addition of arachidonic acid restored VSV growth only partially. Continued growth of HEp-2 cells in DM resulted in a shift in fatty acyl chain composition of phospholipids. The results indicate a finite role for essential fatty acids in receptor-mediated internalization of virus particles. PMID- 2985586 TI - Unidirectional actions of insulin and Ca2+-dependent hormones on adipocyte pyruvate dehydrogenase. AB - Norepinephrine and epinephrine, in the presence of the beta-adrenergic antagonist propranolol (10(-5) M), stimulated adipocyte pyruvate dehydrogenase at low concentrations but inhibited the enzyme at higher concentrations. The alpha adrenergic agonist, phenylephrine, rapidly stimulated pyruvate dehydrogenase activity in a dose-dependent manner with maximal stimulation observed at 10(-6) M. The stimulation of pyruvate dehydrogenase by phenylephrine was mediated via alpha 1-receptors. Inhibition of pyruvate dehydrogenase by catecholamines was mediated via beta-adrenergic receptors, since the beta-agonist, isoproterenol, and dibutyryl cAMP produced similar effects. Like insulin, alpha-adrenergic agonists increased the active form of pyruvate dehydrogenase without changing the total enzyme activity and cellular ATP concentration. The effects induced by maximally effective concentrations of insulin and alpha-adrenergic agonists were nonadditive. The ability of phenylephrine and methoxamine to stimulate pyruvate dehydrogenase and phosphorylase and to inhibit glycogen synthase was not affected by the removal of extracellular Ca2+. Similarly, the stimulation of pyruvate dehydrogenase and glycogen synthase by insulin was also observed under the same conditions. However, when intracellular adipocyte Ca2+ was depleted by incubating cells in a Ca2+-free buffer containing 1 mM ethylene glycol bis(beta-amino-ethyl ether)-N,N,N' -tetraacetic acid, the actions of alpha-adrenergic agonists, but not insulin, on pyruvate dehydrogenase were completely abolished. Vasopressin and angiotensin II also stimulated pyruvate dehydrogenase in a dose-dependent manner with enhancement of glucose oxidation and lipogenesis. Our results demonstrate that the Ca2+ -dependent hormones stimulate pyruvate dehydrogenase and lipogenesis in isolated rat adipocytes, and the action is dependent upon intracellular, but not extracellular, Ca2+. PMID- 2985587 TI - Intracellular processing of epidermal growth factor and its effect on ligand receptor interactions. AB - When normal human fibroblasts are brought to a steady state with 125I-labeled epidermal growth factor (125I-EGF), greater than 90% of the radioactivity is intracellular. We investigated this material to determine whether the 125I-EGF is intact or degraded. Our results show that 125I-EGF is rapidly processed after internalization and can be resolved into four peaks by native gel electrophoresis. These different forms were isolated and tested for their ability to bind to cell-surface EGF receptors. The first processed form was fully capable of binding to EGF receptors, but the second processed form could not. The third form was a collection of small degradation products. We calculated that at steady state about 60% of internalized "125I-EGF" was in a form still able to bind to EGF receptors. We then investigated the ability of different reported inhibitors of EGF "degradation" to block the processing of EGF. Although inhibitors of cathepsin B (leupeptin, antipain, N alpha-p-tosyl-L-lysine chloromethyl ketone, and chymostatin) were able to inhibit the release of monoiodotyrosine from treated cells in a time- and concentration-dependent manner, they had little effect on the processing step that apparently inactivates 125I-EGF. In contrast, agents that raised intravesicular pH, such as methylamine and monensin, inhibited the initial steps in EGF processing as well as the later steps. Low temperatures inhibited the transfer of 125I-EGF to the lysosomes and inhibited the conversion of EGF to a nonbindable form, but had little effect on the initial processing. We conclude that the intracellular processing of EGF is a multistep process that is initiated prior to lysosomal fusion, involves cathepsin B activity, and requires an acidic pH. In addition, many of the protease inhibitors that have been utilized to investigate the role of EGF degradation in mitogenesis do not block the conversion of EGF to a form that is apparently unable to interact with its receptor. PMID- 2985588 TI - Structural requirements for diacylglycerols to mimic tumor-promoting phobol diester action on the epidermal growth factor receptor. AB - The structural requirements for diacylglycerols to mimic the action of tumor promoting phorbol diesters on the epidermal growth factor (EGF) receptor of A431 human epidermoid carcinoma cells were investigated. Five biological effects were considered: inhibition of high affinity 125I-EGF binding, change in the phosphorylation state of the EGF receptor, inhibition of the EGF-dependent tyrosine phosphorylation of the EGF receptor, inhibition of [3H]phorbol 12 beta, 13 alpha-dibutyrate binding, and stimulation of calcium- and phospholipid dependent protein kinase (C-kinase) in vitro. A marked effect of the acyl chain length, 3-10 carbons, of symmetric sn-1,2-diacylglycerols was observed on their ability to mimic the effect of 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA). sn-1,2-Dipropanoylglycerol did not mimic the effects of PMA, but sn-1,2 didecanoylglycerol potently mimicked PMA action. A correlation was found between the ability of these diacylglycerols to stimulate the activity of C-kinase in vitro and to mimic the effects of PMA on the EGF receptor in intact cells. Analogues of sn-1,2-dioctanoylglycerol in which the 3' hydroxyl group was substituted with hydrogen, thio or chloro moieties were inactive when assayed for their ability to stimulate C-kinase in vitro and mimic PMA action in intact cells. We conclude that the hydroxyl group of a diacylglycerol is vital for the interaction with the phorbol diester receptor. The stringent correlation between the potency of the 11 diacylglycerol analogues tested to modulate C-kinase in vitro and to mimic PMA action in vivo provides strong evidence for the hypothesis that C-kinase plays a central role in the regulation of A431 cell EGF receptors by tumor-promoting phorbol diesters. PMID- 2985589 TI - The role of extracellular calcium in corticotropin-stimulated steroidogenesis. AB - The role of extracellular Ca2+ in the binding of corticotropin (ACTH) to adrenocortical cell receptors as well as in the post-binding events involved in steroidogenesis were investigated. Binding studies using [125I Tyr23,Phe2,Nle4]ACTH (1-38) peptide showed that extracellular Ca2+ is essential not only for the interaction of ACTH with its receptor, but also for continued occupancy of the receptor. In view of the requirement of Ca2+ for binding the hormone to the receptor, the role of Ca2+ in post-receptor events was investigated by covalently attaching the hormone to its receptor by photoaffinity labeling in the presence of Ca2+. Persistent activation of steroidogenesis induced by photoaffinity labeling in the presence of Ca2+ was depressed when cells were incubated in medium containing EGTA but was unaffected when the cells were merely washed and incubated in Ca2+-free medium. In the presence of EGTA, 8 Br-cAMP partially restored persistent activation of steroidogenesis. The concentration of extracellular Ca2+ required for restoring steroidogenesis was 10 fold lower than the concentration of Ca2+ needed for optimal binding of ACTH to its receptor. These results suggest that the primary role of extracellular Ca2+ in the action of ACTH is to facilitate the association of the hormone with its receptor. PMID- 2985590 TI - Amino-terminal processing of mutant forms of yeast iso-1-cytochrome c. The specificities of methionine aminopeptidase and acetyltransferase. AB - Amino-terminal processing in the yeast Saccharomyces cerevisiae has been investigated by examining numerous mutationally altered forms of iso-1-cytochrome c. Amino-terminal residues of methionine were retained in sequences having penultimate residues of arginine, asparagine, glutamine, isoleucine, leucine, lysine, and methionine; in contrast, the amino-terminal methionine residues were exercised from residues of alanine, glycine, and threonine and were partially excised from residues of valine. The results suggest the occurrence of a yeast aminopeptidase that removes amino-terminal residues of methionine when they precede certain amino acids. A systematic search of the literature for amino terminal sequences formed at initiation sites suggests the hypothetical yeast aminopeptidase usually has the same specificity as the amino peptidase from bacteria and higher eukaryotes. Our results and the results from the literature search suggest that the aminopeptidase cleaves amino-terminal methionine when it precedes residues of alanine, glycine, proline, serine, threonine, and valine but not when it precedes residues of arginine, asparagine, aspartic acid, glutamine glutamic acid, isoleucine, leucine, lysine, or methionine. In contrast to the normal iso-1-cytochrome c and in contrast to the majority of the mutationally altered proteins, certain forms were acetylated including the following sequences: acetyl(Ac)-Met-Ile-Arg-, Ac-Met-Ile-Lys, Ac-Met-Met-Asn-, and Ac-Met Asn-Asn-. We suggest yeast contains acetyltransferases that acetylates these mutant forms of iso-1-cytochromes c because their amino-terminal regions resemble the amino-terminal regions of natural occurring proteins which are normally acetylated. The lack of acetylation of closely related sequences suggest that the hypothetical acetyltransferases are specific for certain amino-terminal sequences and that the 3 amino-terminal residues may play a critical role in determining these specificities. PMID- 2985591 TI - Identification of the binding site on cytochrome c1 for cytochrome c. AB - The reagent 1-ethyl-3-(3-[14C]trimethylaminopropyl)carbodiimide (ETC) was used to identify specific carboxyl groups on the cytochrome bc1 complex (ubiquinol cytochrome c reductase, EC 1.10.2.2) involved in binding cytochrome c. Treatment of the cytochrome bc1 complex with 2 mM ETC led to inhibition of the electron transfer activity with cytochrome c. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that both the cytochrome c1 heme peptide and the Mr = 9175 "hinge" peptide were radiolabeled by ETC. In addition, a new band appeared at a position consistent with a 1:1 cross-linked cytochrome c1-hinge peptide species. Treatment of a 1:1 cytochrome bc1-cytochrome c complex with ETC led to the same inhibition of electron transfer activity observed with the uncomplexed cytochrome bc1, but to decreased radiolabeling of the cytochrome c1 heme peptide. Two new cross-linked species corresponding to cytochrome c-hinge peptide and cytochrome c-cytochrome c1 were formed in place of the cytochrome c1-hinge peptide species. In order to identify the specific carboxyl groups labeled by ETC, a purified cytochrome c1 preparation containing both the heme peptide and the hinge peptide was dimethylated at all the lysines to prevent internal cross linking. The methylated cytochrome c1 preparation was treated with ETC and digested with trypsin and chymotrypsin, and the resulting peptides were separated by high pressure liquid chromatography. ETC was found to label the cytochrome c1 peptides 63-81, 121-128, and 153-179 and the hinge peptides 1-17 and 48-65. All of these peptides are highly acidic and contain one or more regions of adjacent carboxyl groups. The only peptide consistently protected from labeling by cytochrome c binding was 63-81, demonstrating that the carboxyl groups at residues 66, 67, 76, and 77 are involved in binding cytochrome c. These residues are relatively close to the heme-binding cysteine residues 37 and 40 and indicate a possible site for electron transfer from cytochrome c1 to cytochrome c. PMID- 2985592 TI - Characterization of glycated proteins by 13C NMR spectroscopy. Identification of specific sites of protein modification by glucose. AB - 13C NMR spectroscopy has been used to characterize Amadori (ketoamine) adducts formed by reaction of [2-13C]glucose with free amino groups of protein. The spectra of glycated proteins were acquired in phosphate buffer at pH 7.4 and were interpreted by reference to the spectra of model compounds, N alpha-formyl-N epsilon-fructose-lysine and glycated poly-L-lysine (GlcPLL). The anomeric carbon region of the spectrum (approximately 90-105 ppm) of glycated cytochrome c was superimposable on that of N alpha-formyl-N epsilon-fructose-lysine, and contained three peaks characteristic of the alpha- and beta-furanose and beta-pyranose anomers of Amadori adducts to peripheral lysine residues on protein (pK alpha approximately 10.5). The spectrum of GlcPLL yielded six anomeric carbon resonances; the second set of three was displaced about 2 ppm to lower shielding of the first and was assigned to the Amadori adduct at the alpha-amino terminus (pK alpha approximately 7.5). The spectrum of glycated RNase was similar to that of GlcPLL, but contained a third set of three signals attributable to modification of active site lysine 41 (pK alpha approximately 8.8). The assignments for RNase were confirmed by analysis of spectra taken at pH 4 and under denaturing conditions. The spectrum of glycated hemoglobin was comparable to that of GlcPLL, and distinct resonances could be assigned to Amadori adducts at amino-terminal valine and intrachain N epsilon-lysine residues. Chemical analyses were performed to measure the relative extent of alpha- and epsilon amino group modification in the glycated macromolecules, and the results were compared with estimates based on integration of the NMR spectra. PMID- 2985593 TI - Kinetics of the acid pump in the stomach. Proton transport and hydrolysis of ATP and p-nitrophenyl phosphate by the gastric H,K-ATPase. AB - Hydrolysis of adenosine 5'-triphosphate (ATP) and p-nitrophenyl phosphate by the hydrogen ion-transporting potassium-stimulated adenosine triphosphatase (H,K ATPase) was investigated. Hydrolysis of ATP was studied at pH 7.4 in vesicles treated with the ionophore nigericin. The kinetic analysis showed negative cooperativity with one high affinity (Km1 = 3 microM) and one low affinity (Km2 = 208 microM) site for ATP. The rate of hydrolysis decreased at 2000 microM ATP indicating a third site for ATP. When the pH was decreased to 6.5 the experimental results followed Michaelis-Menten enzyme kinetics with one low affinity site (Km = 116 microM). Higher concentrations than 750 microM ATP were inhibitory. Proton transport was measured as accumulation of acridine orange in vesicles equilibrated with 150 mM KCl. The transport at various concentrations of ATP in the pH interval from 6.0 to 8.0 correlated well with the Hill equation with a Hill coefficient between 1.5-1.9. The concentration of ATP resulting in half-maximal transport rate (S0.5) increased from 5 microM at pH 6.0 to 420 microM at pH 8.0. At acidic pH the rate of proton transport decreased at 1000 microM ATP. The K+-stimulated p-nitrophenylphosphatase (pNPPase) activity resulted in a Hill coefficient close to 2 indicating cooperative binding of substrate. The pNPPase was noncompetitively inhibited by ATP and ADP; half maximal inhibition was obtained at 2 and 100 microM, respectively. Phospholipase C-treated vesicles lost 80% of the pNPPase activity, but the Hill coefficient did not change. These kinetic results are used for a further development of the reaction scheme of the H,K-ATPase. PMID- 2985594 TI - The iron-sulfur cluster composition of Escherichia coli nitrate reductase. AB - Nitrate reductase from Escherichia coli has been investigated by low-temperature magnetic circular dichroism and electron paramagnetic resonance (EPR) spectroscopies, as well as by Fe-S core extrusion, to determine the Fe-S cluster composition. The results indicate approximately one 3Fe and three or four [4Fe 4S]2+,1+ centers/molecule of isolated enzyme. The magnetic circular dichroism spectra and magnetization characteristics show the oxidized and reduced 3Fe and [4Fe-4S] centers to be electronically analogous to those in bacterial ferredoxins. The form and spin quantitation of the EPR spectra from [4Fe-4S]1+ centers in the reduced enzyme were found to vary with the conditions of reduction. For the fully reduced enzyme, the EPR spectrum accounted for between 2.9 and 3.5 spins/molecule, and comparison with partially reduced spectra indicates weak intercluster magnetic interactions between reduced paramagnetic centers. In common with other Fe-S proteins, the 3Fe center was not extruded intact under standard conditions. The results suggest that nitrate reductase is the first example of a metalloenzyme where enzymatic activity is associated with a form that contains an oxidized 3Fe center. However, experiments to determine whether or not the 3Fe center is present in vivo were inconclusive. PMID- 2985595 TI - 5-Hydroxytryptamine stimulates inositol phosphate production in a cell-free system from blowfly salivary glands. Evidence for a role of GTP in coupling receptor activation to phosphoinositide breakdown. AB - Phosphoinositide breakdown has been linked to the receptor mechanism involved in the elevation of cytosolic Ca2+. In a cell-free system prepared from [3H] inositol-labeled blowfly salivary glands, 5-hydroxytryptamine stimulated the rapid production of inositol phosphates. Within 30 s of hormone addition, there was a 100% increase in inositol trisphosphate formation, a 70% increase in inositol bisphosphate formation, and a 90% increase in inositol monophosphate formation as compared to control homogenates incubated for the same length of time. 5-Hydroxytryptamine did not stimulate inositol or glycerol phosphoinositol formation. Half-maximal activation of inositol phosphate production was obtained with 0.33 microM 5-hydroxytryptamine. Ethylene glycol bis(beta-aminoethyl ether) N',N',N',N'-tetraacetic acid, (EGTA) (0.3 mM) inhibited the basal formation of inositol phosphates and decreased the net accumulation of inositol bisphosphate and inositol trisphosphate due to hormone as compared to homogenates incubated in the absence of added Ca2+. EGTA, however, had little effect on the per cent stimulation of inositol phosphate production due to hormone. In homogenates, ATP, GTP or guanyl-5'-yl imidodiphosphate (Gpp(NH)p) was required for a hormone effect. Gpp(NH)p, unlike ATP or GTP, increased the basal formation of inositol phosphates. In membranes, GTP, Gpp(NH)p, or guanosine 5'-(3-O-thio)trisphosphate (GTP gamma S) sustained a hormone effect whereas ATP was ineffective. GTP did not affect production while Gpp(NH)p and GTP gamma S increased inositol phosphate production. Half-maximal effects of Gpp(NH)p and GTP gamma S on hormone stimulated inositol phosphate formation occurred at 10 microM and 100 nM, respectively. In the presence of 1 microM GTP gamma S, 5-methyltryptamine stimulated inositol phosphate formation within 2 s in membranes. These results indicate that in a cell-free system, GTP is involved in mediating the effects of Ca2+-mobilizing hormones on phosphoinositide breakdown. PMID- 2985596 TI - ATP-dependent protease in bovine adrenal cortex. Tissue specificity, subcellular localization, and partial characterization. AB - Proteolytic activities in bovine adrenocortical mitochondria were investigated using [14C-methyl]casein as a substrate. Washed mitochondria showed a low proteolytic activity at pH 7.5 or 8.2. ATP (5 mM) plus MgCl2 (7.5 mM) stimulated the proteolysis 9 times at pH 8.2. It was further demonstrated unequivocally by various approaches that the ATP-dependent proteolytic activity localizes in mitochondrial matrix. The activity of the solubilized protease was sensitive to N ethylmaleimide, mersalyl acid, phenylmethylsulfonyl fluoride, o-vanadate, m vanadate, vanadyl sulfate, and quercetin but not by oligomycin and ouabain. The ATP-dependent proteolytic activity was eluted at the position of 650,000 daltons on an Ultrogel AcA 22 column as a single symmetrical peak. The gel-filtered enzyme showed high specificity to ATP. GTP and UTP partially substituted ATP. ADP, AMP, tripolyphosphate, alpha, beta-methylene ATP, and beta, gamma-methylene ATP had little or no stimulating activity. ATP did not stimulate the activity in the absence of MgCl2. We measured ATP-dependent proteolytic activities in mitochondrial fractions from several tissues in rat and bovine. Adrenal cortex was one of the tissues of highest activity. In addition, we investigated the effect of adrenal atrophy on the ATP-dependent protease activity in rat adrenal. The ATP-dependent protease activity/adrenal decreased by dexamethasone treatment. The extent of the decrease was similar to that of cytochrome oxidase and succinate dehydrogenase, but smaller than that of cytochrome P-450. PMID- 2985597 TI - A selenomethionine-containing azurin from an auxotroph of Pseudomonas aeruginosa. AB - The production and spectroscopic properties of an L-selenomethionine-containing homolog of Pseudomonas aeruginosa azurin are described. The amino acid substitution was carried out by developing an L-methionine-dependent bacterial strain from a fully functional ATCC culture. Uptake studies monitored using L [75Se]methionine indicated that L-selenomethionine was incorporated into the protein synthetic pathway of Pseudomonas bacteria in a manner analogous to L methionine. Several batches of bacteria were grown, and one sample of isolated and purified selenoazurin (azurin in which methionine was substituted by selenomethionine) was found (by neutron activation analysis) to contain 5.2 +/- 0.8 seleniums/copper. Correspondingly, a residual 0.35 methionines, relative to 6.0 in the native protein, were found by amino acid analysis in this azurin sample. The redox potential and extinction coefficient of this selenoazurin were found to be 333 +/- 1 mV (pH 7.0, I = 0.22) and 5855 +/- 160 M-1 cm-1 at 626 +/- 1 nm, respectively. Visible electronic, CD, and EPR spectra are reported and Gaussian curve fitting to the former spectrum allowed assignment of the selenomethionine Se----Cu(II) transition to a band found at 18034 cm-1, based upon an observed 450 cm-1 shift to the red from the analogous band position in the native protein. The data are consistent with a relatively more covalent copper site stabilizing the reduced, Cu(I), form in the selenoprotein. A role for the methionine as a modulator of the blue copper site redox potential by metal--- ligand back bonding from Cu(I) is discussed in terms of a ligand sphere which limits the valence change at copper to much less than 1 during a redox cycle. PMID- 2985598 TI - Partial covalent structure of the human alpha 2 type V collagen chain. AB - Human cDNA libraries were screened with a cDNA fragment presumably encoding the 3' terminus of a procollagen carboxyl propeptide not identifiable as types I, II, III, or IV by protein sequence or Northern blot hybridization. One clone contained a 1350-base pair insert coding in part for 55 uninterrupted Gly-X-Y triplets. Comparison with the amino acid composition of the COOH-terminal cyanogen bromide (CB) peptides of the alpha 1 and alpha 2 type V collagen chains showed similarity only to the alpha 2(V)CB fragment. To identify the NH2 terminus of the peptide designated by methionine, an additional isolate was sequenced and found to contain a Gly-Met-Pro triplet. Thirty-one amino acids from the NH2 terminus of the alpha 2(V)CB9 fragment were then determined by Edman degradation and found to be identical to those derived from the cDNA clone. The DNA sequence encoding part of the triple helical region establishes for the first time the partial structure of a type V collagen chain. Although comparison of residues 796 1020 of the alpha 2(V) collagenous region with alpha 1 (III), alpha 1(I), and alpha 2(I) shows strong conservation of charged positions, the latter three chains appear considerably more similar to each other than to alpha 2(V). A striking feature of the alpha 2(V) sequence between 918-944 is the absence of proline residues. In the analogous region of alpha 1(I) where this amino acid is also lacking, a flexible site in the rigid triple helical structure of type I collagen has been observed (Hofmann, H., Voss, T., Kuhn, K. and Engel, J. (1984) J. Mol. Biol. 172, 325-343). PMID- 2985599 TI - Characterization by electron paramagnetic resonance and studies on subunit location and assembly of the iron-sulfur clusters of Bacillus subtilis succinate dehydrogenase. AB - Succinate dehydrogenase is a conserved membrane-bound enzyme consisting of two nonidentical subunits: a flavo iron-sulfur protein (Fp) subunit, containing a covalently bound flavin, and an iron-sulfur protein (Ip) subunit. Bacillus subtilis succinate dehydrogenase in wild type bacteria and 12 well characterized succinate dehydrogenase-defective mutants were examined by low temperature EPR spectroscopy to characterize the enzyme and study subunit location and biosynthesis of its iron-sulfur clusters. The wild type B. subtilis enzyme contains iron-sulfur clusters which are analogous to clusters S-1 and S-3 of bovine heart succinate dehydrogenase but with slightly different EPR characteristics. Spins from cluster S-2 were not detectable as in the case of the intact form of bovine heart succinate dehydrogenase. However, dithionite reduction of the B. subtilis enzyme greatly enhanced spin relaxation of the ferredoxin-type cluster S-1, indicating the presence of the cluster S-2. Iron sulfur cluster S-1 was found to be assembled in soluble succinate dehydrogenase subunits in the cytoplasm, but only if full-length Fp polypeptides and relatively large fragments of Ip polypeptides were present. Cluster S-1 was not detected in mutants with soluble mutated Fp polypeptides or in a mutant totally lacking Ip subunit polypeptide. Iron-sulfur clusters S-1, S-2, and S-3 were assembled also when the covalently bound flavin in the Fp subunit was absent. Clusters S-1 and S 3 in the membrane-bound flavin-deficient succinate dehydrogenase were not reduced by succinate but could be reduced by electron transfer from NADH dehydrogenase via the menaquinone pool. PMID- 2985600 TI - Electrostatic interactions during electron transfer reactions between c-type cytochromes and flavodoxin. AB - The interaction of three different c-type cytochromes with flavodoxin has been studied by computer graphics modelling and computational methods. Flavodoxin and each cytochrome can make similar hypothetical electron transfer complexes that are characterized by nearly coplanar arrangement of the prosthetic groups, close intermolecular contacts at the protein-protein interface, and complementary intermolecular salt linkages. Computation of the electrostatic free energy of each complex showed that all were electrostatically stable. However, both the magnitude and behavior of the electrostatic stabilization as a function of solution ionic strength differed for the three cytochrome c-flavodoxin complexes. Variation in the computed electrostatic stabilization appears to reflect differences in the surface distribution of all charged groups in the complex, rather than differences localized at the site of intermolecular contact. The computed electrostatic association constants for the complexes and the measured kinetic rates of electron transfer in solution show a remarkable similarity in their ionic strength dependence. This correlation suggests electrostatic interactions influence electron transfer rates between protein molecules at the intermolecular association step. Comparative calculations for the three cytochrome c-flavodoxin complexes show that these ionic strength effects also involve all charged groups in both redox partners. PMID- 2985601 TI - A gap at a unique location in newly replicated kinetoplast DNA minicircles from Trypanosoma equiperdum. AB - Kinetoplast DNA is a network containing thousands of interlocked minicircles. The minicircles replicate as free molecules after release from the network, and their progeny are then reattached (Englund, P. T., (1979) J. Biol. Chem. 254, 4895 4900). In Trypanosoma equiperdum, some of the newly replicated minicircles which have recatenated to the network contain a single gap. This gap is about 10 nucleotides in size and it is in the newly synthesized strand. Based on several criteria (S1 nuclease digestion, denaturing polyacrylamide gel analysis and DNA sequencing), the gap residues at a unique site on the molecule. This site overlaps the sequence GGGGTTGGTGTAA, which is the only common sequence found in all minicircles, from several different species, which have been examined. PMID- 2985602 TI - Site-specific mutagenesis on a human initiator methionine tRNA gene within a sequence conserved in all eukaryotic initiator tRNAs and studies of its effects on in vitro transcription. AB - We have used oligonucleotide-directed site-specific mutagenesis to generate a mutant human initiator tRNA gene in which the sequence GATCG corresponding to the universal GAUCG found in loop IV of eukaryotic cytoplasmic initiator tRNAs is changed to GTTCG. The mutant tRNA gene has been characterized by restriction mapping and by DNA sequencing. We show that this mutation has no effect on in vitro transcription of the tRNA gene in HeLa cell extracts. Transcripts derived from both the wild type (A54) and the mutant (T54) initiator tRNA genes are processed in vitro to produce mature tRNAs with the correct 5'-and 3'-termini. Fingerprint analysis of in vitro transcripts shows that the mutant RNA has the expected nucleotide change. Modified nucleotide composition analyses on the RNAs show that when A54 is changed to U54, the neighboring nucleotide U55 is modified quantitatively to psi 55 in the in vitro extracts; U54 itself is partially modified to ribo-T. Other modified bases identified in the in vitro transcripts include m1G, m2G, m7G, D, and m5C. PMID- 2985603 TI - Expression in vivo of a mutant human initiator tRNA gene in mammalian cells using a simian virus 40 vector. AB - We have cloned both the wild type (A54) and mutant (T54) human initiator genes described in the preceding paper (Drabkin, H. J., and RajBhandary, U. L. (1985) J. Biol. Chem. 260, 5580-5587) as 141-base pair fragments into the SV40-pBR322 vector pSV1GT3. These vectors were subsequently used to transfect monkey kidney CV-1 cells to obtain recombinant virus stocks carrying each of the initiator tRNA genes. Following infection of CV-1 cells by the recombinant virus stocks, both the wild type and mutant tRNAs are produced in large quantities during a 48-h period. Fingerprint analysis of 32P-labeled tRNAs was used to characterize the tRNAs made in vivo and to show that the sequence AUCG in loop IV of the wild type tRNA is replaced by T psi CG in the mutant tRNA. Modified nucleotide composition analysis of the [32P]tRNAs overproduced in vivo shows that they contain all the modified nucleotides found in human placenta initiator tRNA. Both wild type and mutant initiator tRNAs can be aminoacylated by either mammalian or Escherichia coli methionyl-tRNA synthetases. Furthermore, the mutant tRNA can be easily separated from the endogenous monkey initiator tRNA by RPC-5 column chromatography. PMID- 2985604 TI - Characterization of the ileS-lsp operon in Escherichia coli. Identification of an open reading frame upstream of the ileS gene and potential promoter(s) for the ileS-lsp operon. AB - The preceding paper presents evidence for the co-transcriptional expression of the ileS and lsp genes in Escherichia coli. To identify the promoter for the ileS lsp operon, we have determined the nucleotide sequence of an 1.8-kilobase DNA fragment between the rpsT and IleS genes. The sequence data have revealed an open reading frame, designated gene X, which encodes a polypeptide with 312 amino acid residues. Both in vivo and in vitro expressions of the x gene result in the synthesis of a soluble protein with an apparent Mr of 35,000. The x gene is transcribed in the same direction as that of the ileS-lsp operon and opposite to that of the upstream adjacent rpsT gene. No transcription termination sequence can be discerned in the intercistronic region between the x and ileS genes. DNase I footprinting experiment revealed a RNA polymerase binding site at 170-151 base pairs upstream of the x gene. PMID- 2985605 TI - Folylpoly-gamma-glutamate synthetase-dihydrofolate synthetase. Cloning and high expression of the Escherichia coli folC gene and purification and properties of the gene product. AB - The Escherichia coli gene for folylpolyglutamate synthetase-dihydrofolate synthetase was localized to plasmids pLC22-45, 24-31, and 28-44 of the Clarke Carbon E. coli colony bank (Clarke, L., and Carbon, J. (1976) Cell 9, 91-99) by screening the bank by replica mating with an E. coli folC mutant. The folC gene was subcloned from pLC22-45 and inserted into a high copy number plasmid containing the lambda replication control region under the control of the temperature-sensitive cI857 repressor and into a high expression plasmid containing the lambda PL promoter and the cI857 repressor. The folC structural gene was located on a 1.52-kilobase PvuI fragment, sufficient to code for a protein of maximum Mr 55,000. E. coli transformants containing the recombinant plasmids, when induced by culturing at 42 degrees C, had folylpolyglutamate synthetase and dihydrofolate synthetase levels that were 100- to 400-fold higher than in wild type strains and which represented up to 4% of the soluble cell protein. The E. coli folylpolyglutamate synthetase-dihydrofolate synthetase has been purified to homogeneity from the transformants. Both activities are catalyzed by a single protein of Mr 47,000. Some kinetic properties of the enzymes and a new spectrophotometric method for assaying dihydrofolate synthetase activity are described. PMID- 2985606 TI - Role of the nuclear envelope in synthesis, processing, and transport of membrane glycoproteins. AB - The outer nuclear membrane is morphologically similar to rough endoplasmic reticulum. The presence of ribosomes bound to its cytoplasmic surface suggests that it could be a site of synthesis of membrane glycoproteins. We have examined the biogenesis of the vesicular stomatitis virus G protein in the nuclear envelope as a model for the biogenesis of membrane glycoproteins. G protein was present in nuclear membranes of infected Friend erythroleukemia cells immediately following synthesis and was transported out of nuclear membranes to cytoplasmic membranes with a time course similar to transport from rough endoplasmic reticulum (t 1/2 = 5-7 min). Temperature-sensitive mutations in viral membrane proteins which block transport of G protein from endoplasmic reticulum also blocked transport of G protein from the nuclear envelope. Friend erythroleukemia cells and NIH 3T3 cells differed in the fraction of newly synthesized G protein found in nuclear membranes, apparently reflecting the relative amount of nuclear membrane compared to endoplasmic reticulum available for glycoprotein synthesis. Nuclear membranes from erythroleukemia cells appeared to have the enzymatic activities necessary for cleavage of the signal sequence and core glycosylation of newly synthesized G protein. Signal peptidase activity was detected by the ability of detergent-solubilized membranes of isolated nuclei to correctly remove the signal sequence of human preplacental lactogen. RNA isolated from the nuclear envelope was highly enriched for G protein mRNA, suggesting that G protein was synthesized on the outer nuclear membrane rather than redistributing to nuclear membranes from endoplasmic reticulum before or during cell fractionation. These results suggest a mechanism for incorporation of membrane glycoproteins into the nuclear envelope and suggest that in some cell types the nuclear envelope is a major source of newly synthesized membrane glycoproteins. PMID- 2985607 TI - Superinduction of cytochrome P1-450 gene transcription by inhibition of protein synthesis in wild type and variant mouse hepatoma cells. AB - Inhibition of protein synthesis superinduces transcription of the cytochrome P1 450 gene in Hepa 1c1c7 mouse hepatoma cells. The superinduced transcription rate is 10-15-fold higher than the maximal rate of transcription induced by the known inducer 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) alone. Superinduction is maximal within 30-40 min and remains maximal for at least 90 min. Cytochrome P1 450 mRNA is the same length in TCDD-induced and superinduced cells. Superinduction does not occur in variant cells in which TCDD-receptor complexes bind weakly to nuclei and which do not transcribe the cytochrome P1-450 gene in response to TCDD. Inhibition of protein synthesis does not alter several properties of TCDD-receptor complexes. The results imply that a second control mechanism modulates the action of the TCDD-receptor complex in regulating cytochrome P1-450 gene transcription. PMID- 2985608 TI - Binding of apolipoprotein A-I and A-II after recombination with phospholipid vesicles to the high density lipoprotein receptor of luteinized rat ovary. AB - To determine the apolipoprotein specificity of high density lipoprotein (HDL) receptor, apolipoprotein A-I (apo-AI) and apolipoprotein A-II (apo-AII) purified from high density lipoprotein3 (HDL3) were reconstituted into dimyristoyl phosphatidylcholine vesicles (DMPC) and their ability to bind to luteinized rat ovarian membranes was examined. Both 125I-apo-A-I.DMPC and 125I-apo-A-II.DMPC were shown to bind to ovarian membranes with Kd = 2.87 and 5.70 micrograms of protein/ml, respectively. The binding of both 125I-apo-A-I.DMPC and 125I-apo-A II.DMPC was inhibited by unlabeled HDL3, apo-A-I.DMPC, apo-A-II.DMPC, apo-C I.DMPC, apo-C-II.DMPC, apo-C-III1.DMPC, and apo-C-III2.DMPC, but not by DMPC vesicles, bovine serum albumin.DMPC or low density lipoprotein. Since the binding labeled apo-A-I.DMPC and apo-A-II.DMPC was inhibited by the DMPC complexes of apo C groups, the direct binding of 125I-apo-C-III1.DMPC was also demonstrated with Kd = 9.6 micrograms of protein/ml. In addition, unlabeled apo-A-I.DMPC, and apo-A II.DMPC, as well as apo-C.DMPC, inhibited 125I-HDL3 binding. 125I-apo-A-I, 125I apo-A-II, and 125I-apo-C-III1 in the absence of DMPC also bind to the membranes. These results suggest that HDL receptor recognizes apolipoprotein AI, AII, and the C group and that the binding specificity of the reconstituted lipoproteins is conferred by their apolipoprotein moiety rather than the lipid environment. In vivo pretreatment of rats with human chorionic gonadotropin resulted in an increase of 125I-apo-A-I.DMPC, 125I-apo-A-II.DMPC, and 125I-apo-C-III1.DMPC binding activities. However, no induction of binding activity was observed when the apolipoprotein was not included in DMPC vesicles. An examination of the equilibrium dissociation constant and binding capacity for 125I-apo-A-I.DMPC and 125I-apo-A-II.DMPC after human chorionic gonadotropin treatment revealed that the increase in binding activity was due to an increase in the number of binding sites rather than a change in the binding affinity. These results further support our contention that apo-A-I, apo-A-II, and the apo-C group bind to HDL receptor. In conclusion, the HDL receptor of luteinized rat ovary recognizes apolipoproteins A-I, A-II, and the C group but not low density lipoprotein, and the binding is induced by human chorionic gonadotropin in vivo. PMID- 2985609 TI - The DNA restriction endonuclease of Escherichia coli B. I. Studies of the DNA translocation and the ATPase activities. AB - Electron microscopic examination of DNA intermediates formed by the restriction endonuclease of Escherichia coli B revealed supercoiled loops that are presumably formed during an ATP-dependent DNA translocation process in which the enzyme remains bound to the recognition site while tracking along the DNA helix to a cleavage site. The rate of DNA translocation during this process is at least 5000 base pairs/min at 37 degrees C. Even after all cleavages have been completed, complexes are seen that contain terminal loops or loop plus tail structures. During this later phase of the reaction, ATP is hydrolyzed at a rate which is dependent upon the size of the largest possible loop (or loop plus tail); this ATP hydrolysis can be terminated by one double-strand cleavage within the loop region between the recognition site and the terminus. To explain these results, it is hypothesized that after cleavage the enzyme cycles between a tracking (and possibly back-tracking) mode which is fueled by ATP hydrolysis and a relatively long static period in which ATP hydrolysis does not occur. While tracking, the enzyme would be bound both to the recognition site and to a distal site but, while static, the enzyme would be bound only at the recognition site of nonlooped molecules. This post-nuclease phase of the reaction is hypothesized to reflect a reaction whereby the enzyme initially scans DNA molecules before making a strand cleavage. PMID- 2985610 TI - The DNA restriction endonuclease of Escherichia coli B. II. Further studies of the structure of DNA intermediates and products. AB - The DNA intermediates and final products formed by the Type I restriction endonuclease, EcoB, were further characterized. DNA cleaved on only one strand (hemi-restricted DNA) contains gaps of approximately 70-100 nucleotides, while the fully restricted products contain 3'-single-stranded tails averaging approximately 70-100 nucleotides for each strand cleaved. The gaps and tails are formed with the release of an equal number of nucleotides as small oligonucleotides that are soluble in acid. After purification, neither the hemi restricted nor the fully restricted DNAs are cleaved again by EcoB. There is no apparent specificity for which strand of a duplex is initially cleaved by EcoB, nor is there specificity with respect to the composition of the 3'-terminal nucleotide formed on the DNA or the 3'- or 5'-terminal nucleotides of the acid soluble oligonucleotides released during DNA cleavage. The structure formed at the 5' terminus of the DNA product which blocks phosphorylation by T4 polynucleotide kinase remains unknown, but its removal with phage lambda exonuclease allows at least some reutilization of recognition sites by EcoB as well as phosphorylation of the newly formed 5' termini. To explain the complex mechanism of this enzyme, it is suggested that the unidentified 5'-tails prevent wasteful rerestriction from occurring, whereas the 3'-single-stranded tails create DNA which, when nonhomologous to chromosomal DNA, cannot be rescued because such tails are not substrate for DNA polymerases. However, when homologous chromosomal DNA exists, the randomly cleaved large fragments with these tails can easily be assimilated by recA-mediated genetic recombination, thus stimulating DNA exchange between related organisms. PMID- 2985611 TI - Modification and processing of internalized signal sequences of prolipoprotein in Escherichia coli and in Bacillus subtilis. AB - We have cloned the Escherichia coli lipoprotein structural gene (lpp) into a shuttle vector and studied its expression in both E. coli and in Bacillus subtilis. Using in vitro gene fusion techniques, the lpp gene was placed under the control of the promoter for the erythromycin-resistance (ery) gene. This fusion gene directed the synthesis of Braun's prolipoprotein which can be subsequently processed into the mature lipoprotein. In addition to the prolipoprotein, two ery-lpp hybrid proteins containing a 45- and a 22-amino acid extension preceding the NH2 terminus of prolipoprotein, respectively, are also synthesized in E. coli. The synthesis of these three proteins appears to involve the utilization of three distinct translation initiation sites. In B. subtilis, only two proteins are synthesized, the hybrid protein with a 45-amino acid extension and the prolipoprotein. In both E. coli and B. subtilis, the precursor forms of the hybrid proteins are lipid-modified, and they are processed to mature lipoprotein in vivo. These results indicate that internalized signal sequence containing the prolipoprotein modification and processing site (Leu-Ala-Glys-Cys) can function normally and permit the modification of hybrid proteins to lipid modified precursors which can be subsequently processed by the globomycin sensitive prolipoprotein signal peptidase. PMID- 2985612 TI - Regulation of cytochrome P-45011 beta gene expression by adrenocorticotropin. AB - Recombinant DNA clones corresponding to 11 beta-hydroxylase cytochrome P-450 (P 450(11)beta) have been identified in a bovine adrenocortical cDNA library. These clones, pB11 beta-1 and pB11 beta-2, hybridize to at least three RNA species, 7.2, 6.2, and 4.3 kilobases in length, present in the adrenal cortex. All three RNA species directed cytochrome P-450(11)beta synthesis in an in vitro translation system. Expression of the cytochrome P-450(11)beta gene is tissue specific in that these transcripts were not detected in liver, heart, kidney, or corpus luteum. In cultured bovine adrenocortical cells, adrenocorticotropin (ACTH) or dibutyryl cAMP increased the concentration of cytochrome P-450(11)beta transcripts. This increase appears to be at least partially due to de novo transcription as shown by the action of actinomycin D which effectively blocked the ACTH-induced increase in cytochrome P-450(11)beta RNA level. Furthermore, cycloheximide administered prior to or along with ACTH resulted in the blockage of any new transcription of the cytochrome P-450(11)beta gene as evidenced from the level of RNA. Thus, in addition to potential effects on RNA stabilization, a primary action of ACTH in the regulation of the level of cytochrome P-450(11)beta may be to mediate the induction of an unidentified adrenocortical protein which is required for the activation of cytochrome P-450(11)beta gene expression. These results, together with previous studies (Kramer, R.E., Rainey, W.E., Funkenstein, B., Dee, A., Simpson, E. R., and Waterman, M. R. (1984) J. Biol. Chem. 259, 707 713) that showed increased intracellular levels of cAMP upon treatment of bovine adrenocortical cell cultures with ACTH, suggest a role for cAMP-mediated events in the regulation of cytochrome P-450(11)beta gene expression. PMID- 2985613 TI - Microheterogeneity of microtubule-associated proteins, MAP-1 and MAP-2, and differential phosphorylation of individual subcomponents. AB - High molecular weight microtubule-associated proteins 1 and 2 (MAP-1 and MAP-2), prepared by copolymerization with tubulin, were electrophorectically separated into three and two major subcomponents, respectively, using 5% sodium dodecyl sulfate-polyacrylamide gels. By two-dimensional gel electrophoresis, all five MAP components were shown to possess a pI of around 5. Four of these proteins, MAP 1A, MAP-1C, MAP-2A, and MAP-2B, present in comparable amounts, were iodinated after electrophoretic separation and analyzed by two-dimensional peptide mapping. With both trypsin and V8 protease, almost identical patterns were obtained from MAP-2A and MAP-2B. MAP-1A and MAP-1C, too, gave similar digestion patterns, although some differences were noted. Incubation with [gamma-32P]ATP demonstrated that endogeneous protein kinase activities phosphorylated individual subcomponents at different rates. MAP-2A, the highest labeled component, was phosphorylated 2.5-fold compared to MAP-2B both in the presence and the absence of cAMP. Labeling of MAP-1 subcomponents was 4 times less than that of MAP-2A in the absence and 16 times less in the presence of cAMP. 32P-labeled MAP-2A and MAP 2B bands were indistinguishable by one-dimensional peptide mapping, as were the three MAP-1 bands. For both MAP-1 and MAP-2 subcomponents, cAMP induced phosphorylation at new molecular sites. Incubation of radiolabeled microtubule proteins with 1 mM ATP effected, upon electrophoresis, a clear shift of MAP-2A and MAP-2B bands to positions of higher apparent molecular weights, while only slightly affecting MAP-1 bands. PMID- 2985614 TI - Rat liver glutathione S-transferases. Construction of a cDNA clone complementary to a Yc mRNA and prediction of the complete amino acid sequence of a Yc subunit. AB - Using polysomal immunoselected rat liver glutathione S-transferase mRNAs, we have constructed cDNA clones using DNA polymerase I, RNase H, and Escherichia coli ligase (NAD+)-mediated second strand cDNA synthesis as described by Gubler and Hoffman (Gubler, U., and Hoffman, B. S. (1983) Gene 25, 263-269). Recombinant clone, pGTB42, contained a cDNA insert of 900 base pairs whose 3' end showed specificity for the Yc mRNA in hybrid-select translation experiments. The nucleotide sequence of pGTB42 has been determined, and the complete amino acid sequence of a Yc subunit has been deduced. The cDNA clone contains an open reading frame of 663 nucleotides encoding a polypeptide comprising 221 amino acids with a molecular weight of 25,322. The NH2-terminal sequence deduced from pGTB42 is in agreement with the first 39 amino acids determined for a Ya-Yc heterodimer by conventional protein-sequencing techniques. A comparison of the nucleotide sequence of pGTB42 with the sequence of a Ya clone, pGTB38, described previously by our laboratory (Pickett, C. B., Telakowski-Hopkins, C. A., Ding, G. J.-F., Argenbright, L., and Lu, A.Y.H. (1984) J. Biol. Chem. 259, 5182-5188) reveals a sequence homology of 66% over the same regions of both clones; however, the 5'- and 3'-untranslated regions of the Ya and Yc mRNAs are totally divergent in their sequences. The overall amino acid sequence homology between the Ya and Yc subunits is 68%, however, the NH2-terminal domain is more highly conserved than the middle or carboxyl-terminal domains. Our data suggest that the Ya and Yc subunits of the rat liver glutathione S-transferases are products of two different mRNAs which are derived from two related yet different genes. PMID- 2985615 TI - src kinase catalyzes the phosphorylation and activation of the insulin receptor kinase. AB - When a partially purified insulin receptor preparation immobilized on insulin agarose is incubated with [gamma-32P]ATP, Mn2+, and Mg2+ ions, the receptor beta subunit becomes 32P-labeled. The 32P-labeling of the insulin receptor beta subunit is increased by 2-3-fold when src kinase is included in the phosphorylation reaction. In addition, the presence of src kinase results in the phosphorylation of a Mr = 125,000 species. The Mr = 93,000 receptor beta subunit and the Mr = 125,000 32P-labeled bands are absent when an insulin receptor deficient sample, prepared by the inclusion of excess free insulin to inhibit the adsorption of the receptor to the insulin-agarose, is phosphorylated in the presence of the src kinase. These results indicate that the insulin receptor alpha and beta subunits are phosphorylated by the src kinase. The src kinase catalyzed phosphorylation of the insulin receptor is not due to the activation of receptor autophosphorylation because a N-ethylmaleimide-treated receptor preparation devoid of receptor kinase activity is also phosphorylated by the src kinase. Conversely, the insulin receptor kinase does not catalyze phosphorylation of the active or N-ethylmaleimide-inactivated src kinase. Subsequent to src kinase-mediated tyrosine phosphorylation, the insulin receptor, either immobilized on insulin-agarose or in detergent extracts, exhibits a 2-fold increase in associated kinase activity using histone as substrate. src kinase mediates phosphorylation of predominantly tyrosine residues on both alpha and beta subunits of the insulin receptor. Tryptic peptide mapping of the 32P-labeled receptor alpha and beta subunits by high pressure liquid chromatography reveals that the src kinase-mediated phosphorylation sites on both receptor subunits exhibit elution profiles identical with those phosphorylated by the receptor kinase. Furthermore, the HPLC elution profile of the receptor auto- or src kinase catalyzed phosphorylation sites on the receptor alpha subunit are also identical with that on the receptor beta subunit. These results indicate that: the src kinase catalyzes tyrosine phosphorylation of the insulin receptor alpha and beta subunits; and src kinase-catalyzed phosphorylation of insulin receptor can mimic the action of autophosphorylation to activate the insulin receptor kinase in vitro, although whether this occurs in intact cells remains to be determined. PMID- 2985617 TI - The determination of the oxygen content of ampoules and vials of medical biological preparations filled with an inert gas. PMID- 2985616 TI - Substrate-dependent differences in growth and biological properties of fibroblasts and epithelial cells grown in microcarrier culture. AB - Normal diploid human fibroblasts and first passage monkey kidney epithelial cells were examined for growth and metabolic activity on microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells cm2 of surface area) on the glass microcarriers made from glass and on microcarriers made from DEAE-dextran. The cells grew to a higher density (cells/cm2 of surface area) on the glass microcarriers than they did on the DEAE dextran microcarriers and morphological differences were observed between the cells growing on the two substrates. On the DEAE-dextran microcarriers, the cells were much more resistant to protease-mediated detachment than were the cells on the glass microcarriers. In these respects, the cells grown on the glass microcarriers were similar to cells grown in conventional monolayer culture. Interestingly, the cells grown on the DEAE-dextran microcarriers expressed higher levels of proteolytic enzyme activity than the cells grown on the glass microcarriers. Substrate-dependent differences in prostaglandin production also occurred--both in unstimulated cells and in cells stimulated with 12-0 tetradecanoyl phorbol acetate. The unstimulated cells on the glass microcarriers produced slightly higher levels of three different prostaglandins than did the cells on the DEAE-dextran microcarriers. However, after stimulation the levels were much higher in the DEAE-dextran microcarrier cultures than in the glass microcarrier cultures. In contrast to these results, there was no significant, substrate-dependent difference in the production of infectious herpes simplex virus. Taken together, these findings suggest that when commercially-useful cells such as normal fibroblasts and epithelial cells are grown in large quantities on microcarriers, the nature of the substrate may have a profound effect on the growth and physiology of the cells. They also suggest that when microcarriers are used, unexpected results based on preliminary work in conventional monolayer culture may be obtained. PMID- 2985618 TI - The development of mechanical strength of surgically anastomosed arteries sutured with Dexon. AB - The strength of surgically anastomosed arteries of the rat sutured with Dexon thread is studied. The abdominal aortas and the carotid arteries were severed, sutured and then the wounds were closed and the animal healed. After a specific period of time up to 13 months, the vessels were taken out and tested in uniaxial loading condition. The stress-strain relationship of the vessels was measured, and then the vessels were pulled to failure. It was found that the strength of the anastomosis was the lowest in about 4 months. In the first day, the force at failure was about the same as that of the control. Then the strength decreased with time, until a minimum was reached in 4-6 months. The tensile force to failure was about 25% of the control for the carotid artery and 49% of the control for the abdominal aorta. The corresponding values of the tensile stress at failure were 17 and 11%, respectively. The different percentages of forces and stresses were caused by the thickening of the vessel wall in the neighborhood of the suture line in the healing process. After 4-6 months, the strength gained again. At 13 months, the strength of the anastomosis was about the same as that of the control. The stretch ratio at failure was approximately constant through all periods. PMID- 2985619 TI - GABAA and GABAB receptors in detrusor strips from guinea-pig bladder dome. AB - The effects of various GABAA and GABAB receptor agonists and antagonists on electrically induced contractions of detrusor strips from guinea-pig urinary bladder dome were investigated by using both supra and submaximal parameters of stimulation. In supramaximally stimulated preparations GABA (1 mM) inhibited amplitude of contractions. This effect was mimicked, to a lesser degree, by the selective GABAB receptor agonist, (+/-)-baclofen (0.1 mM). Exposure to (+/-) baclofen reduced markedly the effects of a subsequent challenge with GABA. The GABAA receptor agonists, muscimol (0.3 mM) and homotaurine (1 mM), produced a slight inhibition of contractions and reduced the effects of a subsequent challenge with GABA. The selective GABAA receptor antagonist, picrotoxin (0.1 mM), had a slight, but significant, antagonistic effect toward GABA, but had no effect against (+/-)-baclofen. GABA inhibition of supramaximally stimulated contractions was partly reduced by previous exposure to atropine (3 microM) or to the putative P2-purinoreceptor antagonist, reactive blue 2 (0.3 mM) as well as by desensitization of P2-purinoreceptors produced by the stable ATP analogue beta gamma-methylene ATP (APPCP). GABA inhibition was unaffected by phentolamine (0.2 microM), propranolol (0.3 microM) or hexamethonium (10 microM). The inhibition produced by atropine plus reactive blue 2 or APPCP desensitization was additive or more than additive. In submaximally stimulated preparations GABA (0.01-1 mM) produced a transient, concentration related enhancement of amplitude of contractions. This effect was mimicked by either muscimol (0.3 mM) or homotaurine (1 mM) but not by (+/-)-baclofen (0.1 mM). A cross desensitization could be observed between the effects of muscimol or homotaurine on one hand and GABA on the other, but not between (+/-)-baclofen and GABA. Picrotoxin (0.03-0.1 mM) produced a concentration dependent antagonism of a noncompetitive type against the excitatory effect of GABA in submaximally stimulated preparations. Previous exposure to either atropine (3 microM), phentolamine (0.2 microM) or hexamethonium (10 microM) failed to affect GABA induced enhancement of submaximally stimulated contractions. On the other hand the effects of GABA were reduced by reactive blue 2 (0.1-0.9 mM) or by desensitization of P2 purinoreceptors. In preparations exposed to tetrodotoxin (TTX, 0.3 microM), field stimulation induced contractions are attributable to a direct excitation of smooth muscle cells. Under these conditions GABA (1 mM) was ineffective, indicating that, in the absence of TTX, it affects the excitability of neural elements in the bladder wall.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985620 TI - 5-Aminovaleric acid interactions with GABAA and GABAB receptors in guinea-pig ileum. AB - The potential interaction of 5-aminovaleric acid (5-AVA) on GABAA and GABAB receptors was investigated on the guinea-pig isolated ileum myenteric plexus preparation. In the unstimulated preparation 5-AVA (0.1-3 mM) produced a transient contraction which was abolished by previous exposure to picrotoxin (0.1 mM), atropine (3 microM) or tetrodotoxin (0.3 microM). Cross desensitization was observed between the contractile effects of 5-AVA and GABA, while a previous exposure to (+/-)-baclofen did not affect 5-AVA induced contractions. 5-AVA antagonized the relaxant effect of (+/-)-baclofen in unstimulated preparations. 5 AVA (1 mM) had no effect on amplitude of twitches in supramaximally stimulated preparations while GABA produced a concentration related inhibition. In the presence of 5-AVA (1 mM) the concentration response curve to GABA was shifted to the right without a reduction of the maximal effect attainable. These observations indicate that 5-AVA interacts with both GABAA and GABAB receptors in guinea-pig ileum. The concentrations required to observe a GABAA effect are of the same order as those which are effective in producing a blockade of GABAB mediated responses. PMID- 2985621 TI - Tubular structures in S49 mouse lymphoma are regulated through in vivo host-cell interaction and in vitro interferon treatment. AB - Malignant S49 mouse lymphoma cells that grow in suspension culture demonstrate in their cytoplasm characteristic tubular structures. These structures also appear in immunogenic, substrate-adherent variants of S49 cells that grow in culture. Upon transfer of both cell types into nude mice, the tubular structures of the adherent variants (and not the suspension-growing cells) undergo a profound alteration whereby their tubular components disappear and clusters of viruslike particles appear. These very closely resemble, on morphological grounds, precursors of B-type retroviruses. This specific in vivo interaction between the host and the S49 variant can be mimicked in culture by treatment of these cells for 24 h with 500 U/ml of mouse interferon. The suspension-growing S49 cells are unresponsive to interferon in this respect. Immunohistochemical analysis reveals that both tubular structures and the viruslike particles represent stages in the morphogenesis of mouse mammary tumor virus. A working hypothesis is advanced relating the regulation of the tubular system to the impaired tumorigenic potential of adherent S49 cells in syngeneic Balb/c hosts. PMID- 2985622 TI - The cercarial glycocalyx of Schistosoma mansoni. AB - Cercariae, the freshwater stage of Schistosoma mansoni infectious to man, are covered by a single unit membrane and an immunogenic glycocalyx. When cercariae penetrate the host skin, they transform to schistosomula by shedding tails, secreting mucous and enzymes, and forming microvilli over their surface. Here the loss of the glycocalyx from cercariae transforming in vitro was studied morphologically and biochemically. By scanning electron microscopy, the glycocalyx was a dense mesh composed of 15-30 nm fibrils that obscured spines on the cercarial surface. The glycocalyx was absent on organisms fixed without osmium and was partially lost when parasites aggregated in their own secretions before fixation. By transmission electron microscopy, a 1-2 microns thick mesh of 8-15-nm fibrils was seen on parasites incubated with anti-schistosomal antibodies or fixed in aldehydes containing tannic acid or ruthenium red. Cercariae transformed to schistosomula when tails were removed mechanically and parasites were incubated in saline. Within 5 min of transformation, organisms synchronously formed microvilli which elongated to 3-5 microns by 20 min and then were shed. However, considerable fibrillar material remained adherent to the double unit membrane surface of schistosomula. For biochemical labeling, parasites were treated with eserine sulfate, which blocked cercarial swimming, secretion, infectivity, and transformation to schistosomula. Material labeled by periodate oxidation and NaB3H4 was on the surface as shown by autoradiography and had an apparent molecular weight of greater than 10(6) by chromatography. Periodate NaB3H4 glycocalyx had an isoelectric point of 5.0 +/- 0.4 and was precipitable with anti-schistosomal antibodies. More than 60% of the radiolabeled glycocalyx was released into the medium by transforming parasites in 3 h and was recovered as high molecular weight material. Parasites labeled with periodate and fluorescein-thiosemicarbazide and then transformed had a corona of fluorescence containing microvilli, much of which was shed onto the slide. Material on cercariae labeled by lodogen-catalyzed iodination was also of high molecular weight and was antigenic. In conclusion, the cercarial glycocalyx appears to be composed of acidic high molecular weight fibrils which are antigenic and incompletely cleared during transformation. PMID- 2985623 TI - Transforming growth factor-beta modulates the high-affinity receptors for epidermal growth factor and transforming growth factor-alpha. AB - The epidermal growth factor (EGF) receptor mediates the induction of a transformed phenotype in normal rat kidney (NRK) cells by transforming growth factors (TGFs). The ability of EGF and its analogue TGF-alpha to induce the transformed phenotype in NRK cells is greatly potentiated by TGF-beta, a polypeptide that does not interact directly with binding sites for EGF or TGF alpha. Our evidence indicates that TGF-beta purified from retrovirally transformed rat embryo cells and human platelets induces a rapid (t 1/2 = 0.3 h) decrease in the binding of EGF and TGF-alpha to high-affinity cell surface receptors in NRK cells. No change due to TGF-beta was observed in the binding of EGF or TGF-alpha to lower affinity sites also present in NRK cells. The effect of TGF-beta on EGF/TGF-alpha receptors was observed at concentrations (0.5-20 pM) similar to those at which TGF-beta is active in promoting proliferation of NRK cells in monolayer culture and semisolid medium. Affinity labeling of NRK cells and membranes by cross-linking with receptor-bound 125I-TGF-alpha and 125I-EGF indicated that both factors interact with a common 170-kD receptor structure. Treatment of cells with TGF-beta decreased the intensity of affinity-labeling of this receptor structure. These data suggest that the 170 kD high-affinity receptors for EGF and TGF-alpha in NRK cells are a target for rapid modulation by TGF-beta. PMID- 2985624 TI - Identification and localization of immunoreactive forms of caldesmon in smooth and nonmuscle cells: a comparison with the distributions of tropomyosin and alpha actinin. AB - Caldesmon is an F-actin cross-linking protein of chicken gizzard smooth muscle whose F-actin binding activity can be regulated in vitro by Ca2+-calmodulin (Sobue, K., Y. Muramoto, M. Fujita, and S. Kakiuchi, 1981, Proc. Natl. Acad. Sci. USA, 78:5652-5655). It is a rod-shaped, heat-stable, F-actin bundling protein and is the most abundant F-actin cross-linking protein of chicken gizzard smooth muscle presently known (Bretscher, A., 1984, J. Biol. Chem., 259:12873-12880). We report the use of polyclonal antibodies to caldesmon to investigate its distribution and localization in other cells. Using immune blotting procedures, we have detected immunoreactive, heat-stable forms of caldesmon in cultured cells having either approximately the same apparent polypeptide molecular weight as gizzard caldesmon (120,000-140,000) or a substantially lower molecular weight (71,000-77,000). Through use of affinity-purified antibodies in indirect immunofluorescence microscopy, we have localized the immunoreactive forms to the terminal web of the brush border of intestinal epithelial cells and to the stress fibers and ruffling membranes of cultured cells. At the light microscope level caldesmon is distributed in a periodic fashion along stress fibers that is coincident with the distribution of tropomyosin and complementary to the distribution of alpha-actinin. PMID- 2985625 TI - Topoisomerase II is a structural component of mitotic chromosome scaffolds. AB - We have obtained a polyclonal antibody that recognizes a major polypeptide component of chicken mitotic chromosome scaffolds. This polypeptide migrates in SDS PAGE with Mr 170,000. Indirect immunofluorescence and subcellular fractionation experiments confirm that it is present in both mitotic chromosomes and interphase nuclei. Two lines of evidence suggest that this protein is DNA topoisomerase II, an abundant nuclear enzyme that controls DNA topological states: anti-scaffold antibody inhibits the strand-passing activity of DNA topoisomerase II; and both anti-scaffold antibody and an independent antibody raised against purified bovine topoisomerase II recognize identical partial proteolysis fragments of the 170,000-mol-wt scaffold protein in immunoblots. Our results suggest that topoisomerase II may be an enzyme that is also a structural protein of interphase nuclei and mitotic chromosomes. PMID- 2985626 TI - Localization of topoisomerase II in mitotic chromosomes. AB - In the preceding article we described a polyclonal antibody that recognizes cSc 1, a major polypeptide component of the chicken mitotic chromosome scaffold. This polypeptide was shown to be chicken topoisomerase II. In the experiments described in the present article we use indirect immunofluorescence and immunoelectron microscopy to examine the distribution of topoisomerase II within intact chromosomes. We also describe a simple experimental protocol that differentiates antigens that are interspersed along the chromatin fiber from those that occupy restricted domains within the chromosome. These experiments indicate that the distribution of the enzyme appears to be independent of the bulk chromatin. Our data suggest that topoisomerase II is bound to the bases of the radial loop domains of mitotic chromosomes. PMID- 2985628 TI - Serum-independent regulation of initiation of DNA synthesis relating to temperature-sensitive defect in rat 3Y1tsD123 fibroblasts and its compensation by simian virus 40. AB - Randomly proliferating 3Y1tsD123 cells are arrested in G1 phase within 24 h after a shift up to 39.8 degrees C (temperature arrest), yet the density-arrested cells (prepared at 33.8 degrees C) enter S phase at 39.8 degrees C with serum stimulation, with or without preexposure to 39.8 degrees C for 24 h (Zaitsu and Kimura 1984a). When the density-arrested 3Y1tsD123 cells were preexposed to 39.8 degrees C for 96 h, they lost the ability to enter S phase at 39.8 degrees C by serum stimulation and required a longer lag time to enter S phase at 33.8 degrees C by serum stimulation than did the cells not preexposed to 39.8 degrees C. Simian virus 40 induced cellular DNA synthesis at 39.8 degrees C in the density arrested 3Y1tsD123 preexposed to 39.8 degrees C for 96 h. In the absence of serum after a shift down to 33.8 degrees C, the temperature-arrested 3Y1tsD123 cells entered S phase and then divided once. We postulate from these results that (1) the ts defect in 3Y1tsD123 is involved in a serum-independent process. Once this process is accomplished, its accomplishment is invalidated slowly with preexposure to 39.8 degrees C. This and the serum-dependent processes occur in parallel but not necessarily simultaneously. The accomplishment of both (all) processes is required for the initiation of S phase. The density-arrested 3Y1tsD123 cells have accomplished the serum-independent process related to the ts defect, but have not accomplished serum-dependent processes. In case of the temperature-arrested 3Y1tsD123 cells, the reverse holds true. The lag time for entry into S phase depends on the preparedness for the initiation of DNA synthesis (on the extent of accomplishment of each of all processes required for entry into S phase). (2) To induce cellular DNA synthesis, simian virus 40 stimulates directly the serum-independent process. However, we do not rule out the possibility that simian virus 40 stimulates serum-dependent processes simultaneously. PMID- 2985627 TI - Electron microscopic characterization of chick embryonic skeletal muscle proteoglycans. AB - In this article, proteoglycans from embryonic chick leg muscle are quantitatively and qualitatively compared with day 8 high density cell culture cartilage proteoglycans by electron microscopy of proteoglycan-cytochrome c monolayers. The visualized proteoglycan profiles were separated into four categories according to shape, size, and complexity. The two major categories were further characterized by lengths of core proteins, lengths of side projections, and distance between side projections. Two large proteoglycans are identifiable in spread leg muscle preparations. One group has a core protein (mean length of 205 nm) from which extend long thin side projections that we interpret to be groups of chondroitin sulfate glycosaminoglycans with a mean length of 79 nm. This large chondroitin sulfate proteoglycan is the only type found in muscle cultures as determined both biochemically in the past and now by electron microscopy and is referred to as muscle proteoglycan. The second large proteoglycan has a mean core protein length of 250 nm and side projections that are visibly shorter (mean length of 38 nm) and thicker than those of the muscle proteoglycan. This group is referred to as the mesenchymal proteoglycan since its biosynthetic origin is still uncertain. We compare these two profiles with the chick cartilage chondroitin sulfate proteoglycan that has a mean core protein length of 202 nm and side projections with a mean length of 50 nm. The data presented here substantiate the earlier biochemical characterization of these noncartilage proteoglycans and establish the unique structural features of the muscle proteoglycan as compared with the similar profiles of the cartilage and mesenchymal proteoglycans. PMID- 2985629 TI - Two receptor classes for epidermal growth factor on pheochromocytoma cells, distinguishable by temperature, lectins, and tumor promoters. AB - Rat pheochromocytoma cells (clone PC12) display cell surface receptors for both nerve growth factor (NGF) and epidermal growth factor (EGF) and therefore provide a useful model system with which to study the role of these receptors in the regulation of proliferation and differentiation. In this paper PC12 cells are demonstrated to possess two classes of EGF receptors, a high-affinity class with 7,600 sites per cell and an apparent dissociation constant (Kd) of 0.05 nM, and a low-affinity class with 62,000 sites per cell and a Kd of 14.1 nM. These findings are contrary to literature data (Huff et al., 1981; Vale and Shooter, 1983) but can be explained in part by differences in experimental conditions. Binding studies at 37 degrees C compared with room temperature demonstrated similar affinities of both classes, but during prolonged incubation at 37 degrees C, the binding capacities of both classes decreased. Furthermore the high-affinity class was sensitive to lectins, such as concanavalin A (Con A), and to the tumor promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Both compounds caused a decrease of the affinity of the high-affinity class without affecting the low-affinity class. At high concentrations of Con A or TPA, a decrease of the apparent number of binding sites of the low-affinity class was also observed. The similarities between the characteristics of EGF binding and NGF binding in PC12 cells are striking and will be discussed. PMID- 2985630 TI - Abortive transformation of temperature-sensitive mutants of rat 3Y1 cells by simian virus 40: effect of cellular arrest states on entry into S phase and cellular proliferation. AB - Four temperature-sensitive (ts) mutants of rat 3Y1 fibroblasts, representing independent complementation groups, cease to proliferate predominantly with a 2n DNA content, at the restrictive temperature (39.8 degrees C) (temperature arrest) or at the permissive temperature (33.8 degrees C) at a confluent cell density (density arrest) (Ohno et al., 1984). We studied the temperature- or the density arrested cells of these mutants infected with simian virus 40 (SV40) or its mutants affecting large T or small t antigen with respect to kinetics at 39.8 degrees C of entry into S phase and cellular proliferation. Three mutants, 3Y1tsD123, 3Y1tsF121 and 3Y1tsG125, expressed T antigen and entered S phase at 39.8 degrees C from both the arrested states after infection with either wild type, tsA mutants, or a .54/.59 deletion mutant of SV40, whereas in the density arrested 3Y1tsH203, expression of T antigen and entry into S phase were inefficient and ts. Following the WT-SV40 induced entry into S phase, the temperature-arrested 3Y1tsD123 detached from the substratum with no detectable increase in cell number, whereas the density-arrested ones completed a round of the cell cycle and then detached. 3Y1tsF121 and 3Y1tsG125 in the both arrested states proliferated through more than one generation. 3Y1tsF121 and 3Y1tsG125 in the density-arrested state infected with tsA mutants once proliferated and then ceased to increase in number as the percentage of T-antigen positive population decreased. These results suggest that wild-type and tsA-mutated large T antigens are able to overcome the cellular ts blocks of entry into S phase in the 3 ts mutants of 3Y1 cells in both the arrested states, and that small t antigen is not required to overcome the blocks. It is also suggested that cellular behaviors subsequent to S phase (viability, mitosis, and proliferation in the following generations) depend on cellular arrest states, on traits of cellular ts defects, and on the duration of large T antigen expression. PMID- 2985631 TI - Enterocytic differentiation and glucose utilization in the human colon tumor cell line Caco-2: modulation by forskolin. AB - The human colon cancer line Caco-2 exhibits after confluency a concomitant increase of glycogen accumulation and an enterocytic differentiation. The purpose of this work was to investigate whether forskolin (FK), an activator of adenylate cyclase, would induce a permanent glycogenolysis and, if so, whether it would result in modifications of the differentiation pattern of the cells. FK activates adenylate cyclase in Caco-2 cells with an ED50 of 7 X 10(-6)M. Three different treatment protocols with FK (10(-5)M) were applied: 1) the cells were treated during all the time in culture (20 days); 2) the treatment was started after confluency; 3) the treatment was interrupted after confluency. The presence of FK results in a permanent stimulation of cAMP accumulation (10 to 20 fold the basal values) and in a permanently reduced glycogen content (30 or 50% of the control values). The rates of glucose consumption are increased three and five fold in protocols 1 and 3 respectively. These metabolic changes are associated with morphological changes (tightening of the intercellular spaces and shortening of the brush border microvilli) and with a dual inhibition of the activities of brush border hydrolases: a) an inhibition of the post-confluent increase of activity of sucrase, aminopeptidase N and alkaline phosphatase in the brush border enriched fraction; b) an inhibition of the post-confluent increase of activity of sucrase in the cell homogenate. A comparison of the results obtained in each protocol shows that the morphological modifications and the decrease of the enzyme activities in the brush border fraction are regularly associated with an increased cAMP accumulation, whereas the inhibition of the differentiation of sucrase is a direct consequence of the increase in glucose consumption and decrease in glycogen stores. PMID- 2985632 TI - 1 alpha, 25-dihydroxyvitamin D3 specific regulation of growth, morphology, and fibronectin in a human osteosarcoma cell line. AB - The ability of the hormonally active vitamin D metabolite, 1 alpha, 25 dihydroxyvitamin D3, to affect cell growth, morphology and fibronectin production has been examined using the MG-63 human osteosarcoma cell line. Hormone treatment reduced cell growth rate, saturation density and [3H]thymidine incorporation. Inhibition was specific for 1 alpha, 25-dihydroxyvitamin D3 relative to other vitamin D metabolites (1 alpha, 25-dihydroxyvitamin D3 greater than 25 dihydroxyvitamin D3 greater than 24R,25-dihydroxyvitamin D3 greater than D3), antagonized by high concentrations of serum and readily reversed by removal of 1 alpha, 25-dihydroxyvitamin D3 from the culture medium. Hormone treatment also increased cell associated alkaline phosphatase activity up to twofold and altered morphology such that treated cells were more spread out on the culture dish and contained more cytoplasmic processes. Significantly, 1 alpha, 25-dihydroxyvitamin D3 increased cellular and medium concentrations of fibronectin, a glycoprotein known to be involved in cellular adhesiveness. MG-63 cells contain a specific 1 alpha, 25-dihydroxyvitamin D3 receptor which may mediate these responses. PMID- 2985633 TI - The influence of cell population density and serum on the (Na+K)-ATPase of 3T3 cells. AB - The (Na+K)-ATPase of 3T3 cells has been measured as a function of culture density and the type of serum in which the cells were grown. When 3T3 cells were grown in medium containing 10% newborn calf serum their (Na+K)-ATPase activity increased as the culture density increased and the cells became quiescent at densities of about 17.5 X 10(4) cells/cm2. When these cells are subcultured into medium containing 10% foetal bovine serum the activity of the enzyme decreases as the culture density increases and the cells attain final densities of about 5 X 10(4) cells/cm2. PMID- 2985634 TI - Production of a monoclonal antibody directed against the nerve growth factor receptor from sympathetic membranes. AB - Spleen cells from BALB/c mice immunized with a plasma membrane-enriched fraction from rabbit sympathetic ganglia were fused with the mouse myeloma NS1. A hybrid clone was obtained that produced monoclonal antibody directed against the receptor for nerve growth factor (NGF). The antibody, identified as IgG, was able to immunoprecipitate solubilized NGF receptor in the presence or absence of bound NGF. The antibody bound specifically to sympathetic membranes with high affinity but did not affect the binding of 125I-NGF to its receptor in sympathetic or sensory neurons or PC12 cells. PMID- 2985635 TI - A 38 base pair insertion in the pro alpha 2(I) collagen gene of a patient with Marfan syndrome. AB - Abnormalities in type I collagen have been recognized in a number of connective tissue disorders. In the Marfan syndrome, an autosomal dominant condition producing a generalized abnormality in connective tissue, no consistent abnormality has been identified, although one individual has been found to have an elongated pro alpha 2(I) collagen chain [Byers et al, Proc Natl Acad Sci USA 78:7745, 1981]. To determine the nature of the alteration in the gene that produced this abnormality, we studied the pro alpha 2(I) gene from this individual by genomic blotting and gene cloning. Genomic mapping studies detected no abnormalities. However, analysis of the cloned segment of the pro alpha 2(I) collagen gene from the Marfan individual indicates that the gene contains a 38 base pair insertion in an intron near the collagenase cleavage site. Although the relationship of this insertion to the protein abnormality is unclear, it may be a useful marker for the diagnosis of the Marfan syndrome. PMID- 2985636 TI - PDGF stimulates transient phosphorylation of 180,000 dalton protein. AB - Cell-free extracts of platelet-derived growth factor (PDGF) treated, density arrested, quiescent BALB/c-3T3 cells are capable of phosphorylating a 180,000 dalton protein (PP180). The phosphorylation of PP180 was observed in SDS polyacrylamide gel electrophoresis profiles of Nonidet P-40 solubilized cell preparations that had been incubated with [gamma-32P]ATP. When quiescent BALB/c 3T3 cell cultures were incubated at 37 degrees C with PDGF, phosphorylation of PP180 in cell extracts could be detected after a 3-min exposure of the intact cells to PDGF, which was maximal after 10-15 minutes and had diminished by 30-60 min. PDGF stimulation of PP180 phosphorylation also was observed in extracts of cells that had been incubated with PDGF at 4 degrees C; however, in contrast to PDGF exposure at 37 degrees C, the ability of cell extracts to phosphorylate PP180 did not decrease even after 4 hr of cell exposure to PDGF at 4 degrees C. When cells exposed to PDGF at 4 degrees C were transferred to 37 degrees C for 30 min, the ability of cell extracts to phosphorylate PP180 decreased to a nonstimulated level. After cells stimulated by PDGF showed a diminished ability to phosphorylate PP180, immediate restimulation with PDGF did not induce the ability to phosphorylate PP180. Incubation for 11 hr at 37 degrees C was required before readdition of PDGF allowed observable phosphorylation of PP180 in cell extracts, but maximum PDGF stimulation of the phosphorylation of PP180 was found after the cells were incubated for 24 hr in culture conditions. The amount of the stimulation of PP180 phosphorylation was dependent on the concentration of PDGF. The stimulation of DNA synthesis by PDGF was correlated to the phosphorylation of PP180. This phosphorylation activity was not observed in extracts of cells that had been treated with epidermal growth factor (EGF), somatomedin C, insulin, plasma, or fibroblast growth factor (FGF). This novel experimental approach allows the investigation of a PDGF-stimulated phosphorylation activity in relation to the cell cycle and growth regulation. PMID- 2985638 TI - Fe3+-hydroxamate as immobilized metal affinity-adsorbent for protein chromatography. AB - The adsorbent glycinehydroxamate-Sepharose 6B, charged with Fe3+ under specified conditions, is reported. It was used at various pH values for chromatography of the following proteins: lysozyme, cytochrome c, avidin, bovine pancreatic RNase, myoglobin, ovalbumin and human serum albumin. The common naturally occurring amino acids were tested for their interactions with the new sorbent under neutral conditions. PMID- 2985637 TI - Down-regulation of gonadotropin and beta-adrenergic receptors by hormones and cyclic AMP. AB - Loss of gonadotropin receptors in murine Leydig tumor cells and of beta adrenergic receptors in rat glioma C6 cells occurred following exposure of the cells to human chorionic gonadotropin and isoproterenol, respectively. Down regulation of receptors was mimicked in part by other agents that elevated cyclic AMP levels in the cells such as cholera toxin and dibutyryl cyclic AMP. Whereas agonist-mediated receptor loss was rapid and almost total, down-regulation by cyclic AMP was slower and less extensive. Down-regulation of receptors did not appear to be accompanied by loss of the regulatory and catalytic components of adenylate cyclase. Hormone-mediated down-regulation was preceded by desensitization of hormone-stimulated adenylate cyclase. In contrast, there was no evidence that cyclic AMP caused desensitization. Finally, loss of receptors induced either by agonists or cyclic AMP required protein synthesis as cycloheximide inhibited down-regulation. We conclude that down-regulation of receptors in these cells is a complex process involving both cyclic AMP independent and -dependent events. PMID- 2985639 TI - A simple technique for endonuclease mapping of cytomegaloviruses. AB - A simple procedure, suitable for routine epidemiological studies, is described for the differentiation of strains of cytomegaloviruses. Crude DNA from infected cells labelled with 32P was digested with restriction endonucleases and the resultant oligonucleotides were separated by electrophoresis on agarose gels. As few as 10(5) infected cells (i.e. a confluent monolayer of about 1 cm2) provided sufficient DNA for one analysis. PMID- 2985640 TI - Sensitive analytic ELISAs for subclass herpes virus IgG. AB - The subclass distribution of antiviral antibodies to three herpes viruses was studied in a population of healthy blood donors. Subclassification by monoclonal antibodies led to the identification of certain viral IgG patterns. IgG1 appeared to be formed in response to almost all CMV, HSV-1 and VZV infections. A higher frequency of virus-specific IgG3 to CMV and HSV-1 suggested that these infections may be reactivated subclinically more often than VZV. The presence of CMV and VZV IgG4 showed a familial relationship, while IgG4 responses to HSV-1 were common. Persons with IgG4 as the only subclass-reactive antibody to CMV showed cell related reactivities in a high frequency. Patients with leukemias, myelomas and Crohn's disease had a near-normal subclass pattern to the herpes viruses. PMID- 2985641 TI - Abnormal expression pattern of Tac and T9 antigens on in vitro activation of leukemic T cells. AB - Mitogen-induced T-lymphocyte proliferation is dependent on the presence of both interleukin 2 (IL-2) and transferrin, even though resting lymphocytes do not have receptors for either. Recently, it has been reported that IL-2 stimulates T lymphocyte proliferation via IL-2 receptor by induction of transferrin receptor on these cells. Using leukemic T cells as a model of the monoclonal mature T cell, we examined those sequential steps of T-cell activation. Our studies revealed that transferrin receptors do not always appear after IL-2 receptor expression, IL-2 up-regulates the expression of IL-2 receptor but not of transferrin receptor, and IL-2 can initiate DNA synthesis without altering transferrin receptor expression. Thus, the sequential activation steps reported for normal T cells were not observed in the present study on leukemic T cells. These data suggest that there are other pathways to DNA synthesis in leukemic T cells and even in normal T cells. PMID- 2985642 TI - Rescue of IgM, IgG, and IgA production in common varied immunodeficiency by T cell-independent stimulation with Epstein-Barr virus. AB - We previously defined three categories of B-cell defects in common varied immunodeficiency (CVI): failure to produce IgG and IgA in response to T cell dependent (TD) stimulation by Staphylococcus bacteria (Sac) plus pokeweed mitogen or B-cell inducing factor (BIF), failure to produce any immunoglobulin, and failure of Sac-induced proliferation and differentiation. The present study includes the responses of 22 CVI patients to T cell-independent (TI) stimulation by Epstein-Barr virus (EBV). In the majority of patients, EBV-stimulated B cells showed normal proliferation and IgM production. In addition, IgG and IgA production was in the range of that for EBV-stimulated normal cells in many patients. Among 11 patients with no TD production of immunoglobulin of any isotype, two showed normal IgM secretion in response to EBV and five others had significant but subnormal responses. Four patients never had humoral responses despite repeated testing and removal of potentially suppressing T cells and monocytes. Concanavalin A stimulation of the T cells from all the patients tested resulted in the production of B-cell inducing factor at higher levels than for normal donor T cells, as assayed on normal Sac-stimulated B cells. These results show that many cases of B-cell defects in CVI patients involving TD production of IgM, switching to TD production of IgG and IgA, and mitogen responses to Sac are not absolute defects. The B cells will respond normally to some stimuli. PMID- 2985644 TI - Antibody response to cytomegalovirus polypeptides captured by monoclonal antibodies on the solid phase in enzyme immunoassays. AB - Antibodies to different cytomegalovirus (CMV) polypeptide antigens, captured by monoclonal antibodies coated on the solid phase of an enzyme immunoassay test, were analyzed in 42 serum pairs submitted for serodiagnosis of CMV infection. Three CMV antigens, captured on the solid phase by three monoclonal antibodies of different specificities, designated CH92-1, CH65-1, and CH16-1, were glycoproteins A (gA), gC, and gD, respectively; and one antigen, captured by CH23, was a polypeptide with an apparent molecular weight of 150,000, possibly associated with the nucleocapsid. Of these four CMV antigens, gA captured by CH92 1 was most effective in eliciting an antibody response. Antibody to this antigen was present in serum samples at a higher concentration in primary and reactivated infection and persisted longer than did antibody to the other tested antigens. In contrast, antibody to antigen captured by CH23 was at a lower concentration, rose more slowly in infection, and persisted for a shorter time than did antibody to the other antigens. Antibody response to gC and gD was intermediate in concentration and temporal appearance compared with the antibody response to gA and to the polypeptide bound by CH23. An enzyme immunoassay on paired serum samples with the captured glycoproteins as antigen was equal for the detection of current infection to an enzyme immunoassay with the whole CMV antigen from infected cell lysates. Enzyme immunoassays with either the CMV glycoproteins or the whole CMV antigen from infected cell lysates were superior to a complement fixation test with a glycine extract antigen for serodiagnosis of current infection. PMID- 2985643 TI - Enzyme-linked immunosorbent assay for determination of antibodies against herpes simplex virus types 1 and 2 in human sera. AB - A rapid and reproducible enzyme-linked immunosorbent assay (ELISA) is described for determining antibodies in human sera against herpes simplex virus type 1 (HSV 1) and type 2 (HSV-2). The sera were absorbed for 30 min with heterologous virus infected-cell extracts to remove cross-reacting antibodies and then were applied to ELISA plates containing the target antigens, immunoaffinity-purified HSV-1 glycoproteins gC and gD and HSV-2 glycoproteins gD and gF. The absorbance index, defined as the ratio of A414 generated by a serum sample absorbed with a heterologous virus-infected-cell extract versus the A414 of a serum sample absorbed with an uninfected-cell extract, was used to determine the presence or absence of antibodies to HSV-1 and HSV-2. Results of the ELISA for detecting antibodies against HSV-2, when compared with results obtained for the same sera by the microneutralization test, showed an index of overall agreement of 91%. Results of the ELISA for detecting antibodies against HSV-1, when compared with microneutralization test results for sera negative for HSV-2 antibodies but positive for HSV antibodies by ELISA, showed an index of agreement of 99%. PMID- 2985645 TI - Immune response to bovine herpes herpesvirus type 1 infections: virus-specific antibodies in sera from infected animals. AB - The virus specificity of antibodies against bovine herpes virus type 1 was determined with a radioimmunoprecipitation assay and serum collected from natural and experimentally induced infections. By using sequentially collected sera, the development of antibodies to 4 to 5 viral glycoproteins and 11 to 12 nonglycosylated proteins was followed for the first 50 days after infection. The major and most consistent responses in experimentally and naturally infected animals were to four glycoproteins with molecular weights of 102,000, 96,000, 69,000, and 55,000, as well as to a major virion 115,000-molecular-weight nonglycosylated protein. The four glycoproteins were all coprecipitated by a neutralizing monoclonal antibody and were probably involved as target antigens in virus neutralization. Another antigenically unrelated glycoprotein with a molecular weight of 82,000 and a nonglycosylated protein with a molecular weight of 91,000 were also precipitated, but the immune response to these two proteins was transient. Reactivity to gp82 was only weakly detected in serum from naturally infected animals. Contact control animals which did not contract a bovine herpes virus type 1 infection but were exposed to infected animals with signs of severe illness had antibodies which recognized gp102, gp96, gp69 and gp55 as well as p115. These antibodies were present in low amounts and, in contrast to infected animals, did not increase between acute and convalescent sampling. PMID- 2985646 TI - Quantitation of immunoglobulin E antibody to cytomegalovirus by antibody capture enzyme-linked immunosorbent assay. AB - An antibody capture enzyme-linked immunosorbent assay was developed for detection of immunoglobulin E antibody to cytomegalovirus (CMV-IgE). Affinity-purified anti human IgE-coated microtiter plates were used to separate IgE from other classes of antibody in serum. Virus-specific IgE was detected by subsequent incubation with horseradish peroxidase-labeled CMV antigen and substrate. The assay was shown to be very sensitive, since in most positive sera CMV-IgE was still detected at a dilution of 1:5,000. Of 45 patients with primary CMV infection, 43 (96%) were found to produce CMV-IgE. In contrast, CMV-IgE was detected in only 4 (9%) of 44 patients with recurrent CMV infection and in 1 of 144 healthy controls. Furthermore, the level of CMV-IgE in patients with recurrent CMV infection appeared to be lower than that in patients with primary infection. Preliminary examination of successive sera suggested that CMV-IgE is produced somewhat slower than CMV-IgM and -IgA but persists for a shorter period. These results suggest that CMV-IgE may be used as an indicator of primary CMV infection. PMID- 2985647 TI - DNA restriction endonuclease analysis of Mycobacterium bovis and other members of the tuberculosis complex. AB - DNA preparations from 24 New Zealand isolates, two reference strains of Mycobacterium bovis, and one reference strain each of Mycobacterium microti, Mycobacterium africanum, and Mycobacterium tuberculosis were characterized by restriction endonuclease analysis. Twenty-five restriction enzymes were investigated. The clearest differences in M. bovis patterns were obtained with the enzymes BstEII and BclI. These produced four and five different patterns, respectively, for the 24 local isolates. When the results from both enzymes were considered, seven different combinations were obtained. The patterns produced for the two reference strains of M. bovis could be distinguished from each other and also from the patterns produced for the local isolates. All patterns were reproducible and are now being used for typing M. bovis isolates. With either enzyme, the patterns produced for the M. tuberculosis, M. bovis, and M. africanum strains had many features in common, but all the M. bovis patterns were clearly more similar to each other than to the M. tuberculosis patterns. The patterns produced for the M. microti strain were markedly different from those produced for the other species. Restriction endonuclease analysis is clearly a useful method for inter- and intraspecific classifications of the tuberculosis complex. PMID- 2985648 TI - New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki. AB - The genomes of leptospiral field isolates belonging to serogroup Pomona were analyzed and compared with those of type strains by cleavage with restriction endonucleases. This new classification method shows differences among these organisms not indicated by the conventional serological typing method. No differences were observed among isolates from the United States, Canada, and New Zealand. Although all isolates selected for this study had been serologically typed as belonging to serovar pomona, the restriction endonuclease analysis indicates that they belong to serovar kennewicki. kennewicki, a serovar of North American origin, has recently been eliminated from the official serovar list because it was found to be indistinguishable from serovar pomona by the serological method. PMID- 2985649 TI - Examination of uncommon clinical isolates of human adenoviruses by restriction endonuclease analysis. AB - Restriction endonuclease analysis was performed on reference strains of each unknown adenovirus subgenus, in comparison with 40 isolates not identified by our routine methods of neutralization with antisera of species 1 to 8. Several uncommon species would not have been identified initially without the assistance of reference laboratories (species 15, 35, and 37). Other species were identified by comparison with published adenovirus DNA restriction endonuclease patterns or from DNA analysis of reference strains (species 31, 40, and 41). Some isolates could not be matched beyond the level of presumptive adenovirus subgenus. Genomic DNA restriction endonuclease analysis of adenoviruses was useful for the identification of adenovirus isolates in a diagnostic virology laboratory. However, accurate interpretation of results will require more extensive DNA restriction endonuclease fragment analysis of a broader range of adenovirus species and genomic variant strains. PMID- 2985650 TI - Direct appraisal of latex agglutination testing, a convenient alternative to enzyme immunoassay for the detection of rotavirus in childhood gastroenteritis, by comparison of two enzyme immunoassays and two latex tests. AB - During February and March 1984, 207 fecal samples from infants and children with gastroenteritis were tested for rotavirus with four techniques: two enzyme immunoassays (Rotazyme; Abbott Laboratories, North Chicago, Ill., and Enzygnost Rotavirus; Calbiochem-Behring, La Jolla, Calif.) and two latex agglutination tests (Rotalex; Orion Research, Inc., Cambridge, Mass., and Slidex Rota-Kit; Biomerieux). All stool samples were also tested for yeasts and bacterial pathogens. Electron microscopy was used to investigate discrepant results. We found 47% positive samples with Enzygnost-Rotavirus, 38% with Rotazyme, 37% with Slidex Rota-Kit, and 34% with Rotalex. No specimen was found positive by Rotazyme only or Slidex Rota-Kit only. On the contrary, 12 samples which were positive with Enzygnost-Rotavirus only and 3 which were positive with Rotalex only were not confirmed as positive by electron microscopy. Both enzyme immunoassays gave 6% equivocal results; Slidex Rota-Kit gave significantly fewer equivocal results than did Rotalex: 2.9% versus 9.7% (P less than 0.01). The sensitivity and specificity of latex tests compared favorably with that of enzyme immunoassays. Latex agglutination tests can be performed by unskilled personnel and are rapid and relatively cheap. They appear to be very suitable for routine laboratory work and may prove useful for large-scale screening in developing countries. PMID- 2985651 TI - Evaluation of two immunofluorescence assays with monoclonal antibodies for typing of herpes simplex virus. AB - An indirect immunofluorescence assay and a direct immunofluorescence assay were evaluated for typing clinical isolates of herpes simplex virus (HSV). The indirect immunofluorescence assay (Electro-Nucleonics, Inc.) correctly identified 16 HSV type 2 (HSV-2) isolates, but failed to identify 4 of 14 HSV-1 isolates because of background fluorescence and instability of reagents. Forty-nine HSV-1 isolates were correctly typed by direct immunofluorescence assay (Kallestad Laboratories, Inc.), but 1 of 39 HSV-2 isolates did not react with the HSV-2 type specific antibody conjugate. PMID- 2985652 TI - Bone and parathyroid inhibitory effects of S-2(3 aminopropylamino)ethylphosphorothioic acid. Studies in experimental animals and cultured bone cells. AB - S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR 2721) is a radio- and chemoprotective agent which produces hypocalcemia in humans. Intravenous injection of 30 mg/kg WR 2721 in rats and 15 mg/kg in dogs lowers serum calcium by 19 and 25%, respectively. Hypocalcemia in dogs is associated with a fall in serum immunoreactive parathyroid hormone (PTH), which suggests that the mechanism of its hypocalcemic effect is acute hypoparathyroidism. Despite this effect on PTH, in eight chronically parathyroidectomized rats on a low phosphate diet, WR 2721 reduced serum calcium from 9.4 +/- 0.6 to 7.7 +/- 0.5 mg/dl (P less than 0.01) at 3 h. Furthermore, in dogs rendered hypercalcemic by continuous infusion of PTH, WR 2721 reduced serum calcium from 11.0 +/- 0.5 to 10.6 +/- 0.5 mg/dl (P less than 0.01). To determine whether WR 2721 causes hypocalcemia by enhancing the exit of calcium from the circulation or inhibiting its entry, the drug was infused 3 h after administration of 45Ca to rats. WR 2721 did not significantly increase the rate of disappearance of 45Ca from the circulation even though serum calcium fell by 19%. However, serum 45Ca specific activity was higher at 1.5 h (P less than 0.01) and 3 h (P less than 0.05) in rats given WR 2721 than in rats given vehicle alone, which suggests that WR 2721 blocks the entry of nonradioactive calcium into the circulation, presumably from bone. In incubations with fetal rat long bone labeled in utero with 45Ca, 10(-3) M WR 2721 inhibited PTH-stimulated, but not base-line release of 45Ca. Bone resorption by primary culture of chick osteoclasts was inhibited by WR 2721 at concentrations as low as 10(-4) M in the absence of hormonal stimulation. These studies suggest that WR 2721 lowers serum calcium predominantly by blocking calcium release from bone. This acute hypocalcemic effect is at least in part independent of its effect on the parathyroid glands, and is most likely a direct effect of the agent on bone resorption. PMID- 2985653 TI - Isolation and partial characterization of a human alveolar macrophage-derived neutrophil-activating factor. AB - Human alveolar macrophages (AM) were obtained from eight normal volunteers using fiberoptic bronchoscopic lavage to explore potential interrelationships among leukocytes in pulmonary defense against infection. AM placed in monolayer tissue cultures released material into culture supernatants with the capacity to enhance the bactericidal capacity of human neutrophils. Neutrophils preexposed to supernatants killed Pseudomonas aeruginosa from 70 to 90% more efficiently than control cells (P less than 0.02). AM culture supernatants contained this material by 4 h of incubation, and in vitro stimulation of AM cultures with heat-killed P. aeruginosa further increased its production. Gel filtration of AM culture supernatants with a G-50 Sephadex column allowed isolation of a 6,000-D neutrophil-activating factor (NAF) that was resistant to heat (56 degrees C, 30 min). The isoelectric point of NAF, as determined by chromatofocusing, was approximately 7.6. Enzyme digestion of NAF specimens, prepared sequentially by gel filtration and chromatofocusing, demonstrated 50-70% loss of activity after incubations with trypsin, chymotrypsin, and neuraminidase. NAF was only minimally chemotactic and eluted from Sephadex G-50 with particles of a different molecular size than those of AM-derived chemotactic factors (i.e., approximately 10,000 D and less than 500 D). Preincubation of neutrophils with NAF resulted in greater release of superoxide anion upon their subsequent stimulation by either bacterial phagocytosis or by phorbol myristate acetate, as compared with control neutrophils stimulated in a like manner. These studies indicate that human AM secrete a heat-stable, low molecular weight basic protein, with the capacity to enhance oxidative microbicidal activity of neutrophils. PMID- 2985655 TI - Correction of abnormal renal blood flow response to angiotensin II by converting enzyme inhibition in essential hypertensives. AB - In 40-50% of patients with essential hypertension, a high sodium intake does not increase renal blood flow (RBF). These patients have been defined as nonmodulators because sodium intake does not modulate renal and adrenal responsiveness to angiotensin II (AII). To define the role of AII in mediating this altered responsiveness, we assessed the effect of a converting enzyme inhibitor (enalapril) on RBF and its responsiveness to AII in 25 patients with essential hypertension--10 modulators and 15 nonmodulators--and 9 normotensive controls. After 5 d of a 200-meq sodium intake, the nonmodulators did not increase RBF, whereas the normotensives (79 +/- 28 ml/min per 1.73 m2) and modulators (75 +/- 26 ml/min per 1.73 m2) did (P less than 0.025). Arterial blood pressure did not change in the modulators with the salt loading, whereas in the nonmodulators, blood pressure rose (P less than 0.004). After enalapril administration for 66 h, there was a significant difference (P less than 0.01, Fisher Exact Test) in the blood pressure response in the two hypertensive subgroups. In the modulators, there was no change; in the nonmodulators, despite the high salt diet, a blood pressure reduction occurred. In parallel, basal RBF and RBF responsiveness to AII were not changed after converting enzyme inhibition in the normotensive control (n = 9) or the hypertensive modulators (n = 10). Conversely, in the nonmodulators (n = 14), the basal RBF increased significantly (83 +/- 25 ml/min per 1.73 m2; P = 0.01), the increment being indistinguishable from the response to salt loading in normal subjects. Furthermore, renovascular responsiveness to infused AII was also significantly enhanced (P = 0.027) in the nonmodulators, suggesting that enalapril-induced increase in RBF reflected a fall in intrarenal AII levels, and not an increase in prostaglandins or kinins, which would have blunted the renal response to AII. Thus, short-term converting enzyme inhibition corrected abnormalities in sodium-mediated modulation of renal vascular responsiveness to AII. The close quantitative relation of the increase in RBF with sodium loading in normal subjects and modulators, and with converting enzyme inhibition in nonmodulators, viewed in the context of the effectiveness of enalapril only in the latter, and parallel shifts in sensitivity to AII, raises the intriguing possibility that converting enzyme inhibition reversed the failure of the renal blood supply to respond to sodium loading. Thus, converting enzyme inhibitors may reduce blood pressure specifically in this subset of patients with essential hypertension, who are sodium sensitive by way of mechanisms more closely related to local than systemic activity of the renin-angiotensin system. PMID- 2985654 TI - Evidence for carrier-mediated chloride/bicarbonate exchange in canalicular rat liver plasma membrane vesicles. AB - To determine whether anion exchangers might play a role in hepatic bile formation, we looked for the presence of Cl-:OH- and Cl-:HCO3- exchange in highly purified canalicular (c) and basolateral (bl) rat liver plasma membrane (LPM) vesicles. In cLPM vesicles, a pH gradient (7.7 in/6.0 out) stimulated 36Cl- uptake twofold above values obtained during pH-equilibrated conditions (7.7 in = out). When 50 mM HCO3- was also present inside the vesicles, the same pH gradient (7.7 in/6.0 out) resulted in Cl- uptake to levels fourfold above pH- and HCO3- equilibrated controls and two- to threefold above Cl- equilibrium (overshoot). Initial rates of both pH and HCO3- gradient-stimulated Cl- uptake were completely inhibited by 4,4'-diisothiocyano-2,2'-disulfonic acid stilbene (DIDS). A valinomycin-induced K+ diffusion potential (inside positive) also stimulated Cl- uptake in cLPM, but this conductive Cl- pathway was insensitive to DIDS. The DIDS sensitive, pH and HCO3- gradient-stimulated Cl- uptake demonstrated: saturation with Cl- (Km approximately 6.3 mM; Vmax approximately 51 nmol X mg-1 X min-1); partial inhibition by bumetanide (26%), furosemide (33%), probenecid (37%), and 4 acetamido-4'-isothiocyano-2,2'-disulfonic acid stilbene (49%); cis-inhibition by chloride and nitrate but not by sulfate and various organic anions, and independence from the membrane potential. These data demonstrate the presence of an electroneutral Cl-:OH- and Cl-:HCO3- exchanger in rat liver canalicular membranes that favors Cl-:HCO3- exchange. In contrast, no evidence was found for the presence of a Cl-:HCO3- (OH-) exchange system in blLPM vesicles. Furthermore, neither blLPM nor cLPM vesicles exhibited Na+-stimulatable Cl- uptake, indicating the absence of a NaCl co-transport system in either LPM subfraction. These findings are consistent with a functional role for a Cl-:HCO3- (OH-) exchanger in canalicular bile formation. PMID- 2985657 TI - Ultrastructure of rat initial collecting tubule. Effect of adrenal corticosteroid treatment. AB - This study examines the effects of adrenalectomy and physiological replacement of mineralocorticoids and glucocorticoids on the cellular ultrastructure of the rat initial collecting tubule (late distal tubule). Animals were adrenalectomized (ADX) and for 10 d received by osmotic minipump either: vehicle, aldosterone (0.5 micrograms X 100 g-1 X d-1), aldosterone (2.0 micrograms X 100 g-1 X d-1), dexamethasone (1.2 micrograms X 100 g-1 X d-1), or aldosterone (0.5 micrograms X 100 g-1 X d-1) with dexamethasone (1.2 micrograms X 100 g-1 X d-1). Radioimmunoassay revealed that the low dose of aldosterone restored plasma aldosterone to control levels. The higher dose of aldosterone increased plasma levels by threefold. Morphometric techniques were used to measure membrane length of individual principal and intercalated cells in each condition. The basolateral membrane length of principal cells decreased by 35% in ADX animals. Low dose aldosterone replacement (0.5 micrograms X 100 g-1 X d-1) in ADX animals maintained membrane length at control values; at a higher level of aldosterone (2.0 micrograms X 100 g-1 X d-1) membrane length increased by 111% compared with control. Dexamethasone treatment, at a level that restored glomerular filtration rate to normal, had no effect on cellular ultrastructure. Combined aldosterone and dexamethasone replacement had no greater effect on basolateral membrane length than aldosterone alone. The length of the luminal membrane of the principal cell type was not affected by ADX or hormone treatment. Intercalated cell membrane length was not affected by ADX or hormone replacement. Thus, chronic aldosterone levels have an important, selective effect on the basolateral membrane of the principal cell. The correlation between these morphological results and the steroid hormone effects on renal electrolyte excretion, reported in the companion paper (15), suggests that basolateral membrane length is an important factor controlling the rate of sodium and potassium transport by the initial collecting tubule. PMID- 2985658 TI - A comparative study of captopril and enalapril in patients with severe hypertension. AB - Twenty patients with moderate to severe essential hypertension were randomized in a paralleled, double-blind, 22-week study of captopril (ten) and enalapril (ten) following four weeks of hydrochlorothiazide (50 mg/d) baseline treatment. The captopril group was administered 25 mg tid and increased to 100 mg tid, while the enalapril group began with 5 mg bid and increased to 20 mg bid, depending on the patient's blood pressure (BP) response. Methyldopa, 250 mg to 500 mg bid and 1,000 mg bid, was administered to patients in both groups if BP was not adequately controlled. Patients were seen every two weeks for BP monitoring and metabolic evaluation. Each group showed a significant and equal decrease in BP, with the effect being the greatest on the diastolic pressure, supine and upright. Of the 20 patients, ten (four whites and six blacks) required methyldopa for adequate BP control. Four patients, two from each group, developed reversible prerenal azotemia (BUN [blood urea nitrogen] congruent to 50 mg/dL). No other clinical or metabolic side effects were noted. We concluded that captopril and enalapril were equally effective in lowering BP in the dosages given; no racial differences in BP response were noted, although more black patients required the addition of methyldopa for adequate BP control; and both drugs were safe and well tolerated. PMID- 2985656 TI - Role of granulocyte oxygen products in damage of Schistosoma mansoni eggs in vitro. AB - The objectives of this study were to describe the ultrastructure of granulocyte Schistosoma mansoni egg interaction and to determine the role of reduced oxygen products as effectors of cell-mediated damage to the parasite target. Granulocytes attached to the parasites and closely applied their plasma membranes to the microspicules of the egg shell 30 min after mixing in the presence of immune serum. By 4 h, the egg shell was fractured and granulocyte pseudopodia extended toward the underlying miracidium. Granulocyte attachment to eggs resulted in release of O2- (0.30-0.52 nmol/min per 2 X 10(6) cells) and accumulation of H2O2 (0.14-0.15 nmol/min) in the presence of antibody or complement. Granulocytes reduced egg tricarboxylic-acid cycle activity and hatching by 28.3 +/- 0.9 and 35.2 +/- 2.8%, respectively (cell-egg ratio of 1,000: 1). Exogenous superoxide dismutase (10 micrograms/ml) inhibited granulocyte toxicity for egg metabolic activity (3.0 +/- 2.1% reduction in acetate metabolism vs. 28.3 +/- 0.9% decrease in controls without superoxide dismutase, P less than 0.0005) and hatching (12.5 +/- 1.8% reduction, P less than 0.0005), whereas catalase and heparin had no effect. Inhibitors of myeloperoxidase (1 mM azide, cyanide, and methimazole) augmented granulocyte mediated toxicity of egg tricarboxylic-acid cycle activity (44-58% reduction in activity vs. 31 and 35% reduction in controls), suggesting that H2O2 released from cells was degraded before reaching the target miracidium. Oxidants generated by acetaldehyde (2 mM)-xanthine oxidase (10 mU/ml) also decreased egg metabolic activity and hatching by 62.0 +/- 9.0 and 38.7 +/- 7.3%, respectively. Egg damage by the cell-free system was partially prevented by superoxide dismutase (26.5 +/- 4.2% reduction in egg tricarboxylic-acid activity) and completely blocked by catalase (0% reduction in activity). These data suggest that granulocyte-mediated toxicity for S. mansoni eggs is dependent on release of O2- or related molecules. These oxygen products, unlike H2O2, may readily reach the target miracidium where they may be converted to H2O2 or other microbicidal effector molecules. PMID- 2985659 TI - Functional organization in cortical barrels of normal and vibrissae-damaged mice: a (3H) 2-deoxyglucose study. AB - The large mystacial vibrissae on the faces of rodents have punctate representations in all stations in the central trigeminal pathway, including layer IV of the somatosensory cortex (SmI). The cortical whisker correlates, multicellular units termed barrels, are not present at birth, and damage to the vibrissae during the first postnatal week results in altered adult cytoarchitectonics. The anatomical effects of vibrissae damage in the cortex have been well documented; here, we investigated the functional organization of altered SmI barrels with a high-resolution 2-deoxyglucose (2-DG) technique (Durham et al., '81, J. Neurosci. 1:519). The middle row of vibrissae was cauterized in 1-, 2-, 3-, 4-, or 5-day-old mice, and the animals were allowed to survive to sexual maturity. Various combinations of vibrissae were clipped acutely 24 hours prior to injection of 2-4 mCi of (3H)2-DG. Mice actively explored an empty cage for 60 minutes, stimulating the remaining vibrissae. The mice then were perfused and their brains prepared for paraffin histology and emulsion autoradiography. In tangential sections through layer IV, patterns of neuropil and cell body labeling were analyzed with respect to barrel cytoarchitecture in normal and vibrissae-damaged mice. In both control and experimental animals, patterns of neuropil and cell somata label corresponded exactly to barrel boundaries, whether normal or altered by vibrissae damage. Only those barrels for which vibrissae were intact had high levels of label, with anterior barrels more heavily labeled. Many neurons in the septa between these barrels and the adjacent barrels were labeled also. We found slightly higher neuropil label in the cortical zone corresponding to the damaged zone on the face in animals lesioned at any time. These data indicate that physiological somatotopy in vibrissae-damaged animals matches the anatomical cytoarchitecture. PMID- 2985660 TI - The TORCH syndrome: a clinical review. AB - Several infections contracted in utero result in similar physical and laboratory findings during the neonatal period; the TORCH syndrome encompasses such patients. Skin lesions are a frequent finding and the dermatologist may play an important role in the early evaluation of these infants. In this review we discuss etiology, epidemiology, clinical, laboratory, radiologic, and pathologic findings, differential diagnosis, therapy, course, and prognosis of each of these congenital infections. PMID- 2985661 TI - Papillary adenoma of the nipple: analysis of fifteen new cases. AB - Fifteen cases of papillary adenoma (florid papillomatosis, erosive adenomatosis) of the nipple were compared with forty examples of syringadenoma papilliferum, twenty-four of hidradenoma papilliferum, and one hundred of breast carcinoma metastatic to skin. Clinically, papillary adenoma of the nipple typically occurred in a female patient as an erosion, ulceration, or mass that was sometimes accompanied by serous or bloody discharge from the nipple and was frequently misdiagnosed as Paget's disease. Histologically, there was usually a papillary and adenomatous growth in the corium that connected with the surface and showed a lining of apocrine type secretory epithelium, a sometimes plasmacytic stroma, and horn cysts in the upper corium. Papillary adenoma of the nipple is often histologically misinterpreted as syringadenoma papilliferum, hidradenoma papilliferum, or low-grade adenocarcinoma, but it has received almost no attention in the dermatology literature. PMID- 2985662 TI - CT evaluation of invasive trophoblastic disease. AB - We report the use of dynamic CT for the evaluation of trophoblastic disease of the uterus. In five cases showing evidence of persistent trophoblastic disease after evacuation of a molar gestation, contrast enhancement of the myometrium demonstrated several hypodense foci surrounded by highly enhanced areas. These observations were not found in two cases of successfully evacuated hydatidiform mole without sequela. A case of choriocarcinoma showed a large central nonenhanced lesion with peripheral contrast enhancement. Filling defects in a markedly contrast enhanced lesion probably represent hydatids penetrating the myometrium and should suggest invasive mole. PMID- 2985663 TI - Subdural hematoma in patients with brain tumor: CT evaluation. AB - Chronic subdural hematoma was identified radiographically and verified surgically in two patients with brain tumor, one meningioma, and one glioblastoma multiforme. Over a period of days or weeks, both patients developed dramatic clinical findings probably related to the mass effect from the subdural hematoma in addition to the preexisting tumor. Early awareness of this complication in patients with brain tumor may lead to successful surgical treatment. PMID- 2985664 TI - CT features of an unusual hypervascular lung carcinoma complicating chronic asbestosis related pleural disease. AB - The CT appearance of a hypervascular lung carcinoma occurring in a patient with long-standing extensive changes of asbestosis is reported. The diagnostic difficulties that may arise when bronchogenic carcinoma develops in an area of atelectasis is discussed. PMID- 2985665 TI - Computed tomography of mucin-producing adenocarcinoma of the appendix presenting as a bladder tumor. AB - A case of adenocarcinoma of the appendix manifesting as a bladder tumor is presented. Computed tomography contributed valuable information on the origin and extent of the tumor. PMID- 2985666 TI - Isolation, characterization, and regulation of the prolactin receptor. AB - The prolactin, or lactogenic hormone, receptor has been purified (approximately 80%) from lactating mouse liver and human term placenta by the nondenaturing zwitterionic detergent 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate and a prolactin affinity column. The isolated "core-binding unit" has a molecular weight of 37,000 +/- 2,000 daltons. It retains the specificity for lactogenic hormones and binds prolactin with an affinity (Ka = 2 to 6 X 10(9) M 1) similar to that of the receptor as it occurs in its membranous environment (Ka = 3 to 5 X 10(9) M-1). Whether this "core-binding unit" exists on the cell surface in a cryptic or active form is influenced greatly by its association with other membrane proteins and the concentration of phosphatidylcholine within its local membranous environment. PMID- 2985667 TI - Hormonal action controlling mammary activity. AB - Mammary gland differentiation includes multiplication of cells, activation of genes specific to milk synthesis, and activation of "house-keeping" genes. These events are controlled by multiple hormones, the roles of which are not known in detail. Prolactin induction of milk synthesis is accompanied by accumulation of casein messenger ribonucleic acid resulting from acceleration of casein gene transcription as well as stabilization of messenger ribonucleic acid. Prolactin also favors translation of casein messenger ribonucleic acid. Glucocorticoids amplify and progesterone inhibits prolactin action on transcription of casein genes. PMID- 2985668 TI - Response of dairy cows to sodium bicarbonate and limestone in early lactation. AB - A total of 108 dairy cows at three locations were fed a diet supplemented with sodium bicarbonate and limestone during the first 16 wk of lactation. Complete mixed diets were fed for ad libitum intake and consisted of concentrate and corn silage (60:40, dry matter). The four treatments were: 1) basal diet, 2) basal plus 1.2% sodium bicarbonate, 3) basal plus 1.4% limestone, and 4) basal plus 1.2% sodium bicarbonate plus 1.4% limestone. Dry matter intake was increased for 8 wk by sodium bicarbonate, then decreased from wk 9 to 16. Intake was decreased by limestone alone or in combination with sodium bicarbonate. Sodium bicarbonate increased milk production for 8 wk and 4% fat-corrected milk for 16 wk. Average dry matter intake, milk production, and 4% fat-corrected milk (kg/day) for the 16 wk were: 1) 20.2, 31.1, 27.3; 2) 19.8, 32.0, 28.7; 3) 18.9, 31.4, 27.9, and 4) 18.4, 29.7, 27.2 for diets 1 to 4. Ruminal pH was decreased, and volatile fatty acid concentration was increased by added sodium bicarbonate and appeared to result from increased feed intake. Digestibility of nutrients was unaffected by supplements. Sodium bicarbonate affected regulation of blood and urine pH and concentration of electrolytes more than limestone. Serum magnesium was reduced with sodium bicarbonate, although not beyond normal physiological reference range. PMID- 2985669 TI - Abnormal odontogenesis: report of case. AB - All that can be concluded from the available information is that the lesion was a variation on the spectrum of developmental anomalies without a known etiology or a conclusive diagnosis. This lesion is most probably related to an aborted formation of a maxillary first premolar or immature complex odontoma. More information could have been obtained, if the surgery had been delayed until further development of the lesion and of the patient's dentition had occurred; but this was not feasible, since definitive treatment was required. Further radiographic and clinical follow-up may assist in confirming the suspicion of a displaced maxillary left first premolar, when the patient's dentition is further developed. Since the position of the impacted primary tooth had not changed by the first follow-up examination, it is anticipated that it will not erupt on its own and will require further treatment, possibly in the form of surgical exposure and orthodontic manipulation or surgical removal and space management. PMID- 2985670 TI - [Derepressed mutants of the genetic transfer factor pAP38]. PMID- 2985672 TI - Algorithms for removing recovery-related distortion from auditory-nerve discharge patterns. AB - The probability that a cochlear nerve fiber discharges during some specified time interval depends on both the acoustic stimulus and on refractory effects due to earlier spike discharges. With the objective of separating the stimulus-related effects from refractory-related effects seen in poststimulus-time histograms, various maximum-likelihood estimation schemes have been developed. By modeling auditory-nerve fiber discharges as a self-exciting point process in which the intensity depends on both the time during stimulation and the history of discharge, we have been able to independently verify the likelihood estimates of Gaumond et al. [J. Acoust. Soc. Am. 74, 1392-1398 (1983)] under conditions when the recovery function is known. Secondly by maximizing the likelihood function of the neural event process subject to periodic stimulus constraints we have derived estimates which as the number of stimulus presentations and or stimulus periods increase are free from the effects of both absolute and relative recovery-related distortion. Under conditions when the recovery function is unknown, a recursive algorithm is proposed that yields the joint maximum-likelihood estimates of both the stimulus-and recovery-related components in the response histograms. We state the conditions under which unique maximum-likelihood estimates exist and prove that the recursive algorithm converges to those unique estimates. PMID- 2985671 TI - Oxygen-free radicals and lipid peroxidation in adult respiratory distress syndrome. AB - Activated species of oxygen have been implicated as mediators of some acute lung injury. In adult respiratory distress syndrome (ARDS), polymorphonuclear leukocytes accumulate in the lung and release excessive amounts of O2-derived products into the extracellular environment. The effects of these O2 products on lung tissue are multiple. In particular, they can initiate lipid peroxydation in cellular membranes. Excessive lipid peroxydation in membranes destroys cells such as vascular endothelium. Lipid peroxides are also detrimental to cellular functions. Lipid peroxydation could then play a role in the pathogenesis of ARDS. PMID- 2985674 TI - A central spectrum model for the perception of coloration in filtered Gaussian noise. AB - In this paper we describe a monaural auditory signal-processing model for the perception of coloration. The model gives a central spectrum display of a stationary input signal. The central spectrum level for a nerve fiber tuned to a given frequency is computed as a combination of the average firing rate and the firing synchronized to the center frequency of the nerve. The model incorporates a critical-band filter bank, steady-state representations of the average and synchronized firing rates, and temporal integration. The central spectrum model, when used to process simulated data, accurately predicts the perception of coloration in filtered Gaussian noise. PMID- 2985673 TI - The effects of ipsilateral tone burst stimulus level on the discharge patterns of cat lateral superior olivary units. AB - Discharges were recorded extracellularly from single units localized in the lateral superior olive (LSO) of barbiturate anesthetized cats. The statistical properties of the unit discharges to monaurally presented tone bursts were determined. Increasing the stimulus level of an ipsilaterally presented tone burst produced an increase in the discharge rate and the emergence and growth of a time-locked discharge pattern in the initial portion of the response. The initial time-locked response was transient and was followed by a nontime-locked sustained response. In the sustained portion, increasing stimulus level produced increases in rate and changes in the interspike interval statistics. Average rate and interval statistics were found to be systematically related. LSO units were differentiated on the basis of rate and pattern of their initial discharges into two main types: the fast and slow chopping units. An analysis of the results indicated that some aspects of response type differences may be related to input characteristics rather than to neuron response mechanisms. PMID- 2985675 TI - Effect of coronary artery recanalization on right ventricular function in patients with acute myocardial infarction. AB - The effects of coronary artery recanalization by intracoronary administration of streptokinase on left ventricular function during acute myocardial infarction have received increasing attention in recent years. Although myocardial dysfunction is often more pronounced in the right ventricle than in the left ventricle in patients with acute inferior wall myocardial infarction, the effect of coronary artery recanalization on right ventricular dysfunction has not been previously addressed. Accordingly, in this investigation, 54 patients who participated in a prospective, controlled, randomized trial of recanalization during acute myocardial infarction were studied. Among 30 patients with inferior wall infarction, 19 had right ventricular dysfunction on admission; 11 of these 19 had positive uptake of technetium-99m pyrophosphate in the right ventricle, indicative of right ventricular infarction. Patients with successful recanalization (n = 6) exhibited improved right ventricular ejection fraction from admission to day 10 (26 +/- 7 to 39 +/- 14%, p less than 0.03). However, control patients (n = 6) and patients who did not undergo recanalization (n = 7) also exhibited improvement (20 +/- 7 to 29 +/- 11% [p less than 0.02] and 30 +/- 8 to 40 +/- 6% [p less than 0.03], respectively). Improvement in several other variables of right ventricular dysfunction evolved at an equal rate with the ejection fraction changes. Patients with or without right ventricular infarction improved similarly. These data indicate that the right ventricular dysfunction commonly associated with inferior wall infarction is often transient, and improvement is the rule, irrespective of early recanalization of the "infarct vessel." PMID- 2985676 TI - Pharmacokinetic interactions between digoxin and other drugs. AB - Drug interactions with digoxin are important because of this agent's narrow therapeutic index. Among the drugs that can decrease digoxin bioavailability are cholestyramine, antacid gels, kaolin-pectate, certain antimicrobial drugs and cancer chemotherapeutic agents. In selected patients, antibiotics may enhance digoxin bioavailability by eliminating intestinal flora that metabolize digoxin. Antiarrhythmic drugs, such as quinidine and amiodarone, can markedly increase steady state serum digoxin levels. Certain calcium channel blocking drugs, particularly verapamil, have a similar effect. Potassium-sparing diuretic drugs, such as spironolactone, can alter digoxin pharmacokinetics. Indomethacin may decrease renal excretion of digoxin in preterm infants. Finally, rifampin, an antibiotic used in the treatment of tuberculosis, may lower steady state serum digoxin levels in patients with severe renal disease. Physicians must maintain constant vigilance whenever medications are added to or withdrawn from a therapeutic regimen that includes digoxin. PMID- 2985677 TI - Time course of the increase in airway responsiveness associated with late asthmatic reactions to toluene diisocyanate in sensitized subjects. AB - To understand better the mechanism of the increase in airway responsiveness associated with late asthmatic reactions, we determined the time course of toluene diisocyanate (TDI) effect on airway responsiveness in six sensitized subjects who exhibited a late asthmatic response after TDI exposure (0.018 +/- 0.005 ppm, 30 min) in the laboratory. Airway responsiveness was assessed before TDI exposure and then at 8 hr, 1 day, 1 wk, and 1 mo after TDI exposure. To assess responsiveness we determined the provocative dose of methacholine causing a decrease in FEV1 of 20% (PD20FEV1). The methacholine PD20 decreased from 0.50 mg geometric standard error of the mean (GSEM = 1.54) to 0.06 mg (GSEM = 1.55) (p less than 0.001) at 8 hr after exposure to TDI, was still decreased to 0.15 mg (GSEM = 1.93) (p less than 0.05) at 1 day, returned to 0.26 mg (GSEM = 1.91) (p greater than 0.05) at 1 wk, and returned to 0.43 mg (GSEM = 1.71) at 1 mo, indicating that full recovery occurred within 1 to 4 wk. These results demonstrate that TDI-induced late asthmatic response is associated with a reversible increase in airway responsiveness to methacholine and suggest that the TDI effect is linked to an acute inflammatory response in the airways. PMID- 2985678 TI - Dietary change: prescription vs. goal setting. AB - This investigation assessed the effectiveness of two teaching strategies, dietary prescription (Diet-Directed [D-D]) and individual goal setting (Self-Directed [S D]), on compliance with the U.S. Dietary Goals and Guidelines. College women, 18 to 30 years old, enrolled in a one-credit course that met for 1 hour and 15 minutes weekly for 10 weeks to learn the principles and application of the U.S. Dietary Goals and Guidelines. The D-D group received a prescribed diet; the S-D group learned goal-setting skills; and a Control group received no treatment. Caloric, total fat, and polyunsaturated fat (PUFA) intake decreased significantly in both treatment groups. The D-D group reduced caloric, fat, and PUFA intake by week 5 and maintained those changes for the remainder of the experiment. The S-D group decreased caloric intake by the eighth week and decreased fat and PUFA intake between weeks 5 and 8. Those changes, however, were not maintained for the duration of the experiment. The results demonstrate that the study population made dietary changes in the direction of the U.S. Dietary Goals and Guidelines. Further investigations are required to assess the effectiveness of various teaching strategies on long-term dietary compliance. PMID- 2985679 TI - Effect of age on thyroid hormone physiology and function. PMID- 2985680 TI - Thallium replaces potassium in activation of the (Na+,K+)-ATPase of rat liver plasma membranes. AB - The capacity of thallium to substitute for K+ in activation of (Na+,K+)-ATPase of liver plasma membranes was studied. Our results indicate that T1+ can replace K+ in the activation of the (Na+,K+)-ATPase of liver plasma membranes. In the presence of Na+, similar activation is obtained with T1+ concentrations only 1/10 of those of K+. In all other aspects, the (Na+,K+)- and (Na+,T1+)-ATPases were found to be identical. PMID- 2985681 TI - The effect of substrate and potassium on the inhibitory kinetics of MnCl2 on the enzyme K+-p-nitrophenyl phosphatase in rat brain. AB - The effect of Mn2+, a divalent metal, on the enzyme K+-p-nitrophenyl phosphatase (K+-PNPPase) was studied in rat brain. The metal was found to be a moderate inhibitor of the enzyme, with an I50 of approximately 480 microM. The inhibition was pH dependent, but not temperature dependent. On measurement of the inhibition with varying concentrations of PNPP (1-5 mM), the I50 value remained constant. However, when the inhibition was measured with K+ (5-20 mM), the I50 value increased from 130 microM to 490 microM, suggesting that K+ antagonized the effect of Mn2+. In kinetic studies, Mn2+ inhibited the enzyme in a non competitive manner with respect to PNPP. The Km remained constant (2.9), but the Vmax was decreased from 5.0 to 1.6. However, with respect to K+, the inhibition was competitive, as the concentration for half maximal activation (K0.5) increased from 1.3 to 8.9 mmol l-1 with 1 mM of MnCl2, suggesting that the apparent affinity of K+ for the enzyme was decreased. The apparent Vmax was not affected. The degree of cooperactivity (n) measured as the slope of the Hill plot remained unaltered (1.9 +/- 0.2) over the entire concentration range of MnCl2 tested. PMID- 2985682 TI - Adenosine triphosphatase activities in plasma liver membranes of rats treated with DDT and toxaphene. AB - The effect of exposure to chlorinated insecticides (DDT and toxaphene) on Na+,K+ ATPase, Mg2+-ATPase and Ca2+-ATPase activities of the plasma membrane of hepatocytes was determined. Acute treatment with DDT (200 mg per kg body weight) or toxaphene (110 mg per kg body weight) produced a significant decrease in Na+,K+-ATPase activity (80% and 85%, respectively) 24 h after treatment. DDT also produced a 30% decrease in Mg2+-ATPase and Ca2+-ATPase activity, but toxaphene treatment did not modify these enzymes. The effect of exposure to daily doses of DDT (30 mg per kg body weight) or toxaphene (16.5 mg per kg body weight) for a period of 3.5 months was also studied. Animals were sacrificed at 15-day intervals and results showed that Na+,K+-ATPase activity decreased 80% from the beginning of each treatment and the activity remained low throughout the treatment period. DDT, but not toxaphene, also led to a decrease in Mg2+-ATPase (20%) and Ca2+-ATPase (35%) activity. The low values observed from the beginning remained low throughout the treatment period. We believe that the general mechanism of ATPase inhibition by organochloride compounds could be the result of its interaction with membrane lipid components, although some differences could arise from differences in their spatial structure. PMID- 2985683 TI - Neurotoxicity of diallate and triallate when administered orally or topically to hens. AB - Two allylthiocarbamate herbicides, diallate and triallate, were evaluated for neurotoxicity by oral and topical dosing studies with mature white leghorn hens. Diallate was tolerated for 90 days at topical doses of 40 mg/kg/day and oral doses of 20 mg/kg/day. Reversible ataxia and narcosis occurred at diallate doses of 80 mg/kg/day and higher by either route of administration. Triallate did not elicit signs of neurotoxicity at 300 mg/kg/day topically or 400 mg/kg/day orally. The oral dose, however, resulted in gastrointestinal irritation and severe weight loss, such that dosing was terminated after 25 days. Triallate was tolerated at oral dosages of 90 mg/kg/day and topical doses up to 330 mg/kg/day. PMID- 2985684 TI - Malignant hyperthermia, a surgeon's nightmare. AB - Malignant hyperthermia, a medical emergency occuring during anesthesia, may be triggered by any volatile agent. Once begun, malignant hyperthermia cascades into a fulminating cycle with body temperature rising as high as 109 degrees F. Treatment consists of immediate cessation of surgical procedure and anesthesia. The patient's temperature must be quickly lowered by the use of ice baths, cooled intravenous solutions, and the drug of choice, dantrolene. PMID- 2985685 TI - Effect of short and long-term administration of lisuride in Cushing's disease. AB - To define the role of dopaminergic drugs on pituitary ACTH secretion, we studied the effect of a dopamine-agonist, lisuride, on a group of 9 patients with pituitary dependent Cushing's syndrome. Lisuride (0.4-0.6 mg/day) was administered for two days: in only two out of eight cases the ACTH levels, sampled every 30 min for 6 h (08:00-14:00 h), showed some reduction when compared with the control day. Two of the nonresponders (R.P.; B.F.) and one untested patient (L.E.) were then treated chronically with a similar amount of lisuride per day for 20 days (L.E.) and for 4 months (B.F.; R.P.). One patient (L.E.) showed a clear decrease in urinary free cortisol without clinical improvement; while in the other two cases, followed for a longer period of time, a clinical improvement was observed together with a decrease of the hormonal parameters. In conclusion, lisuride may occasionally reduce ACTH secretion in patients with pituitary dependent Cushing's syndrome, but the acute test fails to identify the patients who would benefit from this treatment. PMID- 2985686 TI - Parallel determination of glucocorticoid receptors in human mononuclear and polymorphonuclear leukocytes after Percoll separation. AB - Circulating human leukocytes are known to possess glucocorticoid receptors, but the data in the literature on the affinity and capacity widely range. We here report our results on the determination of affinity and capacity of glucocorticoid receptors in both mononuclear and polymorphonuclear leukocytes using a Percoll gradient for separating cells. The mean affinity of mononuclear and polymorphonuclear cells was respectively 4.3 +/- 0.7 and 4.3 +/- 15 nM (mean +/- S.D., n = 6), and the capacity 4135 +/- 1950 and 1810 +/- 832 sites per cell. Previous reports using a Ficoll separation showed a lower affinity under the same conditions of incubation. From these findings we suggest that the separation with Percoll may give values closer to physiological than other separation techniques. PMID- 2985687 TI - Isolated ACTH deficiency with transitory GH deficiency. AB - A nine-year-old girl, who presented with a severe hypoglycemic coma, proved to have isolated ACTH deficiency, a finding previously reported in only two children. On the initial evaluation, before any treatment, GH did not respond to provocative stimuli. On replacement therapy with hydrocortisone, normal linear growth was observed. Repeated testing while on glucocorticoids replacement four years after the initial attack revealed normal GH response to stimulation test. It is suggested that cortisol deficiency was responsible for the severe hypoglycemic coma and subnormal GH response. A similar mechanism is speculated for the normal growth observed in some patients with apparent deficiency of anterior pituitary hormones, including GH. The possibility of permanent ACTH deficiency and transitory GH deficiency following hypophysitis is discussed. PMID- 2985688 TI - Ketanserin without effects on basal anterior pituitary hormone secretion in healthy subjects. AB - The effect of ketanserin, a selective serotonin-2 (5-HT2) receptor blocking agent, on the secretion of anterior pituitary hormones was studied in 4 healthy volunteers. Ketanserin (10 mg) was administered as a slow iv injection and its effect was compared with that of saline. Ketanserin influenced neither the basal plasma levels of HGH, ACTH, TSH, LH or prolactin, nor the plasma levels of T4, T3, cortisol or glucose. Even if a single dose of ketanserin had no hormonal effects, this must also be studied after long term use. PMID- 2985689 TI - Environmentally induced analgesia: an age-related decline in an endogenous opioid system. AB - The purpose of the present study was to examine in rats the relationship between the age-related decline in an opioid receptor system and the function of the endogenous opioid pain-inhibitory system activated by front-paw shock. Results revealed that the analgesia displayed following front-paw shock declined between 5- to 7-month-old, 15- to 17-month-old, and 22- to 24-month-old age groups. The administration of naloxone significantly attenuated the shock-induced analgesia. Thus, the endogenous opioid pain modulatory system activated by front-paw shock (the nucleus raphe alatus and a descending pathway lying within the dorsolateral funiculus of the spinal cord) declines in function with increasing age. This research also confirms that there is a parallel between the age-related decline in the neurochemical indexes of the opioid receptor system and the function of these receptors in producing analgesia in response to aversive stimulation. PMID- 2985690 TI - The effects of ACTH1-24 or cortisol on pulmonary maturation in the adrenalectomized ovine fetus. AB - Pulmonary maturation in 8 ovine fetuses bilaterally adrenalectomized at 98-101 days and infused at term with either ACTH1-24 or cortisol was compared with that in 4 untreated sham-operated controls. Four of the adrenalectomized fetuses were infused intravascularly with ACTH1-24 5 micrograms/h for 84 h before delivery and the other four were infused with cortisol 1 mg/h for 72 h. The high plasma concentrations of immunoreactive ACTH in the adrenalectomized fetuses (2762 +/- 1339 ng/l, mean +/- SD) were not significantly elevated by infusion of ACTH1-24 but were markedly depressed by infusion of cortisol. Distensibility (V40) of the lungs was less than that of controls in both the ACTH1-24-infused and cortisol infused fetuses (1.86 +/- 0.31 ml/g vs 0.62 +/- 0.13 ml/g and 1.27 +/- 0.34 ml/g respectively) but it was significantly greater in the cortisol-infused fetuses compared to those infused with ACTH1-24. The volume of air retained at 5 cm H2O pressure (V5) during deflation was markedly reduced in adrenalectomized fetuses (controls 1.14 +/- 0.52 ml/g vs 0.25 +/- 0.25 ml/g and 0.12 +/- 0.6 ml/g). The wet weight of the lungs and the concentrations of saturated phosphatylcholine in lung tissue and lavage fluid were lower in the adrenalectomized fetuses than in controls but the differences were not significant. It is concluded that infusion of ACTH1-24 at term in adrenalectomized fetuses is probably without effect whereas cortisol enhances distensibility. PMID- 2985691 TI - Feasibility and efficacy of in-home water chlorination in rural North-eastern Brazil. AB - The purpose of this study was to assess the feasibility, acceptability and effect of an in-home water chlorination programme in a rural village. Previous studies at this site showed high levels of faecal coliforms in household water, high diarrhoea rates in children, and enterotoxigenic Escherichia coli and rotaviruses were the most common pathogens isolated from patients. Household water came from a pond and was stored in clay pots. No homes had sanitary facilities. A blind, cross-over trial of treatment of household water with inexpensive hypochlorite by a community health worker was carried out over 18 weeks among 20 families. Water in the clay pots was sampled serially, and symptom surveillance was done by medical students. The programme was generally acceptable to the villagers and no change in water use patterns were apparent. The mean faecal coliform level in the chlorinated water was significantly less than in the placebo treated samples (70 vs 16000 organisms/dl, P less than 0.001). People living in houses receiving placebo treatment had a mean of 11.2 days of diarrhoea per year, and the highest rate of 36.7 was among children less than 2 years old. Diarrhoea rates were not significantly different among the participants while exposed to water treated with hypochlorite. We conclude that a low-cost programme of this type, which utilizes community resources, is logistically feasible, appears to be culturally acceptable in this setting, and can result in a marked reduction in water contamination. The lack of effect on diarrhoea rates suggests that improvement in water quality may affect morbidity only when other variables relating to faecal oral agent transmission are ameliorated at the same time. PMID- 2985692 TI - Down-regulation of Fc receptor expression in guinea pig peritoneal exudate macrophages by muramyl dipeptide or lipopolysaccharide. AB - Expression of Fc receptors on the plasma membrane of guinea pig peritoneal exudate macrophages (PEM) was suppressed to almost one-half of that of the controls by long-term exposure to lipopolysaccharide (LPS) or muramyl dipeptide (MDP) in culture. The effect of the reagents was dose and time dependent, and as little as 0.5 ng/ml LPS or 5 ng/ml MDP was effective for the suppression. The expression of the Fc receptors decreased to 60 to 70% of the control level at 48 hr and to 45 to 50% at 72 hr after incubation of the cells in the presence of LPS or MDP. A Scatchard plot of the binding of 125I-soluble immune complexes (I.C.) to the cells revealed that the decrease in the binding of 125I-I.C. is due to a reduction in the number of Fc receptors on the cell membrane and not to a decreased affinity of the receptors. The membrane protein was radio-labeled with 125I, and the Fc receptors were purified by being bound to insoluble I.C. The specific binding of the 125I-labeled Fc receptors, from the LPS-treated macrophages, to the insoluble I.C. was almost one-half of that from the untreated control cells. SDS-PAGE analysis of the purified 125I-labeled Fc receptors revealed that the major peak of the m.w. 44,000 molecule in the LPS-treated cells was almost one-half of that of the control. Contrary to the effect of LPS or MDP, 72-hr incubation of macrophages with MIF-rich supernatant, cultured from lymph node cells, enhanced the expression of Fc receptors. Macrophages were treated with I.C. for 4 hr at 37 degrees C to remove the Fc receptors from the surface membrane. The reappearance of the receptors on the plasma membrane of the cells was significantly suppressed by LPS and MDP. The effect of LPS on the binding of five murine monoclonal antibodies (Ab) raised against PEM to the macrophage membrane and also that of 125I-wheat germ agglutinin (WGA) or 125I-insulin was studied. The monoclonal Ab were selected for their activity to induce superoxide anion generation in the macrophages, as do I.C., although the binding sites for the monoclonal Ab were not related to Fc receptors. The bindings of the five monoclonal Ab were not affected by exposure of the cells to LPS or MDP. Macrophages treated with the reagents bound as much 125I-insulin or WGA as did the untreated control cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985694 TI - Stimulation of genetic resistance to marrow grafts in mice by interferon alpha/beta. AB - Lethally irradiated mice were infused with syngeneic, H-2 allogeneic, parental strain, or H-2 heterozygous bone marrow cells. They were injected daily with rabbit anti-mouse interferons (IFN)-alpha/beta or gamma or with IFN-alpha/beta. The growth of donor-derived cells was judged 5 days later by measuring splenic incorporation of 5-iodo-2'-deoxyuridine-125I into DNA. Antibodies to IFN alpha/beta, but not to IFN-gamma, weakened genetic (both hybrid and allogeneic) resistance to marrow cell grafts. IFN-alpha/beta stimulated hybrid and allogeneic resistance, the latter even in genetically "poor responder" mice. Mice pretreated with silica, which weakens genetic resistance, were stimulated by IFN-alpha/beta to resist incompatible marrow cell grafts; however, IFN-alpha/beta failed to reverse the effects of antiasialo GM1 serum on marrow graft rejection. IFN alpha/beta did not inhibit the growth of syngeneic marrow cells and did not stimulate resistance to H-2 heterozygous bone marrow cells. We propose that genetic resistance occurs in two discrete steps. In the first step, hemopoietic histocompatibility (Hh) antigens are recognized by one host cell type, and this recognition leads to IFN-alpha/beta secretion by a silica-sensitive cell. In the second step, asialo GM1-positive natural killer cells stimulated by IFN alpha/beta recognize Hh antigens on marrow stem cells and cause rejection. The defects in resistance observed in genetically poor responder mice and in mice treated with silica appear to involve the first step in recognition. The lack of rejection of H-2 heterozygous (Hh-) marrow cells by parental strain mice injected with IFN-alpha/beta indicated that specific Hh recognition is critical in the second step of genetic resistance. PMID- 2985693 TI - Bacterial endotoxin selectively prevents the expression of scavenger-receptor activity on human monocyte-macrophages. AB - Concentrations of bacterial lipopolysaccharide (LPS) as low as 1 ng/ml suppressed the activity of the scavenger receptor on cultured human monocyte-macrophages. In contrast, concentrations of LPS as high as 100 ng/ml had no effect on the activity of the low density lipoprotein (LDL) receptor. LPS and purified forms of the lipid A moiety of LPS were effective in suppressing scavenger receptor activity. However, acid hydrolysis of the labile phosphate group of the native diphosphorylated lipid A to form monophosphoryl lipid A rendered the molecule ineffective in suppressing scavenger receptor activity. LPS at a concentration of 100 ng/ml had no effect on the secretion of apolipoprotein E, phagocytic activity, tumoricidal activity, or the protein content of monocyte-macrophages. We conclude that the active component of LPS that mediates suppression of scavenger receptor activity is diphosphoryl lipid A. PMID- 2985695 TI - Distribution of cells bearing the Tac antigen during ontogeny of human lymphoid tissue. AB - The monoclonal antibody anti-Tac, which binds to the interleukin 2 (IL 2) receptor, was used to identify this antigen in human fetal and adult lymphoid tissue. Liver, spleen, thymus, lymph node, and peripheral blood were examined for Tac-positive cells with the use of frozen sections or cytocentrifuge preparations. The results show that cells in the fetal and neonatal thymus express the Tac antigen; these cells are predominantly located in the medulla. The liver and spleen of both fetus and adult exhibit very few Tac-positive cells. Double staining demonstrates that cells bearing the Tac-antigen stain with Leu-4, an anti-T cell antibody. In adult lymph node tissue, the Tac-bearing cells are predominantly distributed in the interfollicular area, with positive cells also present in the germinal center and mantle zone. The Tac antigen is present on both T and B cells. Few Tac-positive cells are present in the circulating peripheral blood. PMID- 2985696 TI - Functional association of class II antigens with cell surface binding of Epstein Barr virus. AB - A functional role of class II antigen in the binding of Epstein-Barr virus (EBV) was deduced from the study of membrane proteins on Jijoye, an EBV receptor (EBVR) positive B cell line, and its mutant, EBVR-negative daughter cell line, P3HR-1. From gel electrophoresis of radiolabeled microsomal membrane proteins and immunoprecipitates, we identified class II antigen on Jijoye but not on P3HR-1 cells and the presence of Ii on both cell lines. The role of these molecules in EBVR function was tested by antibody blocking of virus adsorption. Anti-p23,30 serum (to class II antigen) was found to block binding of EBV to B lymphoblasts under conditions in which normal rabbit serum, rabbit antiserum to butyrate treated P3HR-1 cells (with ample anti-Ii antibodies), and rabbit anti-p44,12 (to class I antigen and beta 2-microglobulin) serum did not block virus binding. Only one of four commercial monoclonal antibodies (MoAb) to framework epitopes on class II antigens blocked binding of EBV, whereas all four MoAb demonstrated immunofluorescent reactivity with the EBVR+ Raji cells. In previous studies of binding of EBV to hairy leukemic cells, a substantial subpopulation of HLA-DR+, EBVR- cells was identified, in addition to HLA-DR+, EBVR+ cells. These findings were consistent with the view that the HLA-DR complex has a role in the binding of EBV but that other components are also needed for the expression of EBVR function. PMID- 2985697 TI - Ecto-5'-nucleotidase can provide the total purine requirements of mitogen stimulated human T cells and rapidly dividing human B lymphoblastoid cells. AB - The ability of mitogen-stimulated human T cells or rapidly dividing human B lymphoblastoid cells to drive their total purine requirements from inosine 5' monophosphate, inosine, or hypoxanthine was compared. Inosine 5'-monophosphate first must be converted to inosine by the action of the enzyme ecto-5' nucleotidase before it can be transported into the cell; inosine and hypoxanthine, however, can be transported directly. Mitogen-stimulated human peripheral blood T cells were treated with aminopterin to inhibit purine synthesis de novo and to make the cells dependent on an exogenous purine source. Thymidine was added as a source of pyrimidines. Under these conditions, 30 microM inosine 5'-monophosphate, inosine, and hypoxanthine showed comparable abilities to support [3H]thymidine incorporation into DNA or [3H]leucine incorporation into protein at rates equal to that of untreated control cultures. Similar results were found when azaserine was used to inhibit purine synthesis de novo, and thus DNA synthesis. In parallel experiments with the rapidly dividing human B lymphoblastoid cell line WI-L2, treatment with aminopterin (plus thymidine) inhibited the growth rate by greater than 95%. The normal growth rate was restored by the addition of 30 microM inosine 5'-monophosphate, inosine, or hypoxanthine to the medium. However, in similar experiments with cell line 1254, a derivative of WI-L2 which lacks detectable ecto-5'-nucleotidase activity, inosine and hypoxanthine (plus thymidine), but not inosine 5'-monophosphate (and thymidine) were able to restore the growth inhibition due to aminopterin. These results show that the catalytic activity of ecto-5'-nucleotidase is sufficient to meet the total purine requirements of mitogen-stimulated human T cells or rapidly dividing human B lymphoblastoid cells, and suggest that this enzyme may be important for purine salvage when rates of purine synthesis de novo are limited and/or an extracellular source of purine nucleotides is available. PMID- 2985698 TI - Fine specificities of autoantibodies directed against the Ro, La, Sm, RNP, and Jo 1 proteins defined by two-dimensional gel electrophoresis and immunoblotting. AB - Although useful for specific purposes, immunofluorescence, precipitation in agarose gels, and the m.w. estimation of RNA or proteins immunoprecipitated from transformed cells often provide partial or ambiguous definition of autoantibody specificity. We have analyzed organ and cell extracts by one-and two-dimensional electrophoresis together with Western blotting to define the fine specificities of antibodies to the ribonucleoprotein (RNP) antigens Ro, La, Sm, RNP and Jo-1. One-dimensional analysis identified the Ro protein as a 57 kilodalton (kd) protein, although many anti-Ro sera also react with a 50 kd protein. La antisera react with 50 and 43 kd proteins. The 50 kd La protein readily breaks down into 43, 25, and smaller immunoreactive cleavage products. Partial proteolysis of Ro and La proteins in human spleen extracts produces similar immunoreactive products, providing evidence for a common structure. The major immunoreactive Sm antigens defined by human polyclonal antisera and a mouse monoclonal antiserum were doublets of 25/26 and 16/18 kd, whereas anti-RNP sera reacted with a protein of 68 kd. Most Sm-RNP antisera contained antibodies reactive with additional proteins, especially when whole cell extracts were used as a source of antigens. Two-dimensional analysis provided characteristic maps of the antigens. Ro and La were acidic, and La showed a unique set of acidic charge isomers at 50 and 43 kd. Anti-Sm antibodies reacted with discrete dots corresponding to both the acidic and basic regions of the first-dimension (charge) gels, whereas the RNP antigen showed a series of basic charge isomers of 68 kd. Many anti-Sm-RNP sera reacted with other closely spaced proteins of a similar charge and size to the Sm and RNP antigens, suggesting antibody cross-reactivity or reactivity with closely related functional proteins. Although Jo-1 had the same m.w. as the undegraded La antigen, the fingerprints were quite distinctive on two-dimensional electrophoresis. The results of this study indicate how the source and preparation of antigen extracts, as well as protein degradation, influence the m.w. determinations of soluble protein antigens. With these factors taken into account, two-dimensional fractionation with immunoblotting provides a highly discriminating, sensitive, and reproducible method of analysis of autoantibody specificity. This technique can be used to standardize reference antisera and to study protein antigens in normal and abnormal cell and tissue extracts, and could lead to new or more precise correlations with clinical disease. PMID- 2985700 TI - Effects of immune complexes on production by human monocytes of interleukin 1 or an interleukin 1 inhibitor. AB - The objective of these studies was to examine the ability of phorbol myristic acetate (PMA), Fc fragments, and various forms of immune complexes to induce the production by human monocytes of factors stimulatory to chondrocytes or thymocytes. All of these materials were prepared free of detectable contamination with bacterial lipopolysaccharides (LPS) at the level of less than 0.1 ng/ml. Supernatants and lysates from stimulated human monocytes were assayed for their ability to induce collagenase production in cultured rabbit articular chondrocytes or to augment mitogen-induced proliferation of murine thymocytes. The activity detected by these assays exhibited an m.w. of approximately 15,000, and electrophoretic heterogeneity in the pH ranges of 5 to 5.5 and 6.5 to 7.0, characteristics of human interleukin 1 (IL 1) or IL 1-like factors. Monocytes cultured with 2 ng/ml LPS produced chondrocyte and thymocyte stimulatory factors. PMA, Fc fragments, and soluble, precipitated, particulate, or adherent immune complexes were inactive in stimulating the monocytes. However, complement fixation by precipitated immune complexes did generate activity capable of inducing monocytes to synthesize and secrete chondrocyte and thymocyte stimulatory factors. Adherent immune complexes and PMA were biologically active, as evidenced by induction of superoxide generation in the human monocytes. Supernatants from monocytes cultured on adherent immune complexes contained a factor inhibitory to chondrocyte and thymocyte responsiveness. This factor had a m.w. approximately 22,000 and appeared to inhibit specifically IL 1 stimulation, not interleukin 2 stimulation or cell proliferation. It was concluded that PMA, Fc fragments, and various forms of immune complexes in the absence of complement do not induce IL 1 production in human monocytes. However, complement fixation by immune complexes does lead to activation of monocytes to produce IL 1. Monocytes cultured on adherent immune complexes produce an IL 1 inhibitor. PMID- 2985699 TI - Analysis of nephritogenic antigens in human glomerular basement membrane by two dimensional gel electrophoresis. AB - Collagenase digests of GBM were partially purified by column chromatography and analyzed by 2-D gel electrophoresis. Silver staining of 2-D gels showed charge- and size-related heterogeneity of proteins in the 45 to 50 kDa and 25 to 27 kDa regions. These components were transferred to nitrocellulose sheets and reacted with 10 human anti-GBM autoantibodies. Detection of bound anti-GBM autoantibodies to blotted proteins was carried out with peroxidase-labeled goat anti-human IgG and revealed binding predominantly to the cationic (pI 8 to 9.0) 45 to 50 kDa and 25 to 27 kDa components. Positive-staining patterns of blotted proteins were similar with all anti-GBM autoantibodies except that three sera additionally identified neutral (pI 5.5 to 6.5) protein components. One anti-GBM autoantibody, which developed following renal transplantation, lacked reactivity with the most cationic components in the 25 to 27 kDa region. These findings suggest heterogeneity of nephritogenic GBM antigens. The cationic 45 to 50 kDa components were sensitive to reduction, while one neutral 45 to 50 kDa component was resistant; a complex array of 25 to 30 kDa proteins (pI 5.5 to 7.5) were observed by silver staining postreduction. None of the reduced protein components reacted with anti-GBM antibodies, suggesting that epitopes on nephritogenic GBM antigens may be related to disulfide-bonded regions. Although there is variable immunohistochemical reactivity of anti-GBM autoantibodies with the GBM of infant kidneys, 2-D gels of collagenase-digested human infant GBM blotted and reacted with anti-GBM autoantibodies and showed staining patterns similar to that of adult GBM. These studies demonstrate the presence of nephritogenic antigens in the GBM of immature human kidney which are not detectable by immunohistochemical analysis. PMID- 2985701 TI - Human B lymphoblastoid cell lines provide an interleukin 1-like signal for mitogen-treated T lymphocytes via direct cell contact. AB - The B lymphoblastoid cell lines (B-LCL) 8392, SB, 1788, and Daudi provide accessory cell activity for mitogen-treated T cells, whereas the T lines MOLT-4, 8402, CEM, and HSB do not provide this function. Direct cell contact is required for the accessory cell activity, and active lymphocyte growth factors could not be detected in the supernatants of the B-LCL. The B-LCL also present alloantigens to responding T cells, and this response is independent of additional accessory cells. The target for the B-LCL is the responding T cell itself, rather than a minor contaminating population of endogenous accessory cells. This conclusion is based on the finding that, under culture conditions in which T cells do not proliferate in response to PHA, accessory cell activity of the B-LCL is maintained. Paraformaldehyde- or glutaraldehyde-treated B-LCL retain their accessory cell activity at levels of these agents that completely eliminate metabolic activity of the B-LCL, as determined by incorporation of leucine, thymidine, and uridine into macromolecules. This treatment eliminates alloantigen presentation by the B-LCL. T cells treated with IO-4 or with monoclonal anti-T3 antibodies fail to respond to highly purified IL 1, and respond minimally to supra-optimal concentrations of IL 2. Nevertheless, these cells respond maximally to the accessory cell activity of the B-LCL. The IO-4 treated cells or cells exposed to anti-T3 also proliferate in response to TPA. Together, our data suggest that the B-LCL provide an IL 1-like signal for mitogen-treated T cells via direct cell contact, in the absence of detectable soluble IL 1. PMID- 2985702 TI - Plasma cell antigen PC-1 and the transferrin receptor in mouse, rat, and hamster: serologic and biochemical analysis. AB - The plasma cell membrane antigen PC-1 and the receptor for the iron transport protein transferrin are high m.w., developmentally regulated proteins consisting of two similar or identical disulfide-bonded subunits. In this paper, we report the results of a serologic and biochemical analysis of these proteins in various strains of inbred mice, and in rats and hamsters. A monoclonal antibody against the PC-1a allelic product is shown to detect an antigenic determinant on the PC-1 molecule that has the same strain distribution as the antigen previously detected with polyclonal alloantisera. The mouse PC-1 protein was purified from plasma cells of the PC-1a genotype and was used to generate polyclonal rabbit anti-PC-1 antibodies. These antibodies precipitated a homologous protein from plasmacytoma cells derived from PC-1- congenic mice, demonstrating that PC-1b is not a "null" allele. The PC-1b allelic product had a slightly lower apparent m.w. than the PC 1a product, and had a slightly more basic isoelectric point. Rabbit anti-mouse PC 1 antibodies also precipitated a homologous protein from immunoglobulin-secreting cells of rat and hamster origin, but did not show detectable cross-reaction with the transferrin receptor. Disulfide bonding between chains was conserved in both PC-1 and the transferrin receptor in all species examined, but transferrin receptors from mouse cells had a significantly higher apparent m.w. than those of rat, hamster, or human cells. PMID- 2985703 TI - Cyclic AMP-dependent protein kinase in Molt 4b lymphoblasts: identification by photoaffinity labeling and activation in intact cells by vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine (PHI). AB - Molt 4b lymphoblasts have previously been shown to possess a single class of pharmacologically specific, high affinity receptors for vasoactive intestinal polypeptide (VIP). This study further explores the molecular basis for modulation of human lymphocyte function by VIP. Dose-dependent stimulation of adenylate cyclase was observed in Molt lymphoblasts over the range of 0.1 nM to 1 microM VIP. VIP-mediated by guanine nucleotide. Accumulation of intracellular cAMP was observed in the presence of either VIP or the diterpene, forskolin. The effects of these two agonists were synergistic. Two neuropeptides that share sequence homology with VIP were also studied; both peptide histidine isoleucine (PHI) and human pancreatic growth hormone releasing factor (1-44 GHRF) competed for 125I VIP binding to Molt cells. PHI stimulated intracellular cAMP accumulation and demonstrated synergism with forskolin, whereas GHRF had no effect on cAMP. Photoaffinity labeling of 100,000 X G soluble proteins with 8-N3-[32P]cAMP followed by SDS gel electrophoresis demonstrated the presence of cAMP-dependent protein kinases I and II. Cyclic AMP-dependent protein kinase II predominated in the soluble fraction and was the only isozyme observed in particulate fractions. Protein phosphorylation was studied in Molt 4b cells preincubated with [32P]PO43- followed by addition of media alone, 1 microM peptide, or 10 microM forskolin. Cells were lysed and subjected to two-dimensional electrophoresis. Increased phosphorylation of a specific 41,000 Mr protein was observed after addition of forskolin, VIP, or PHI. A much lower concentration of VIP (1 nM) also caused a significant net increase in phosphorylation, which was of a lower magnitude. In contrast, no net effect on protein phosphorylation was seen with GHRF. These data demonstrate the presence of a functional VIP receptor that is linked to the G protein-adenylate cyclase complex. The demonstration of cAMP-dependent protein kinase and of VIP- and PHI-mediated protein phosphorylation in Molt 4b lymphoblasts provides evidence on a molecular level for neuropeptide modulation of human lymphocyte function. PMID- 2985704 TI - Characterization of three monoclonal antibodies which induce and modulate superoxide anion generation in guinea pig macrophages. AB - Three monoclonal antibodies (Ab), termed KY 12, KY 22, and KY 25 and raised against guinea pig macrophages, induced superoxide anion (O2-) generation in the cells. Although each monoclonal Ab bound to macrophages, each had a different pattern of binding to other cell types. In response to each of the Ab, the amount of O2- generated by 5 X 10(5) macrophages was between 0.5 and 0.7 nmol/min and was augmented threefold to fivefold by the addition of F(ab')2 fragments of rabbit Abs to mouse Ig. When macrophages were pretreated with soluble immune complexes (I.C.) prior to stimulation by the monoclonal Ab, the O2- generation stimulated by KY 12 or KY 22 was reduced by more than 70%. In contrast, pretreatment of macrophages with I.C. did not reduce O2- generation in response to KY 25. KY 12 and KY 22 stimulated adenyl cyclase activity in macrophages, but KY 25 did not. Pretreatment of the cells with soluble I.C. did not interfere with the enhancing effect of the monoclonal Ab on adenyl cyclase activity. Pretreatment of macrophages with KY 12 reduced by over 60% of subsequent generation of O2- in response to wheat germ agglutinin, I.C., formyl-methionyl leucyl-phenylalanine, phorbol myristate acetate, KY 22, or KY 25. KY 22 or KY 25 did not suppress the generation of O2- in response to other stimuli. These results suggest that KY 22 and KY 25 activate O2- generation in a manner that differs from that of KY 12. These monoclonal Ab should prove useful in examining the regulation of O2- production. PMID- 2985705 TI - Construction of novel class I histocompatibility antigens by interspecies exon shuffling. AB - Human and mouse class I histocompatibility antigens share considerable structural homology at both the protein and DNA sequence level. This homology has allowed the production of hybrid class I molecules by the reciprocal exchange of DNA sequences corresponding to equivalent domains of HLA-B7 and either H-2Ld or H 2Dd. It is shown that these genes give rise to protein products that are stably expressed on the surface of murine L cells after DNA-mediated gene transfer. These proteins express only those monoclonal antibody-defined H-2 determinants that are expected based on their genetic construction. The molecules have allowed the localization of a number of polymorphic and monomorphic HLA-specific epitopes. In all but one case, expression of an epitope on a domain does not appear to be influenced by the replacement of adjacent human domains with their murine equivalents, suggesting a considerable degree of structural independence of the domains. Cells expressing the hybrid molecules have also been tested as targets for a panel of HLA-B7-specific cytotoxic T cell clones. The results show that the polymorphic determinants recognized by these clones map to the alpha 1 and alpha 2 domains of the HLA-B7 molecule. No evidence for an influence of species-related amino acid sequence differences in the third extracellular domain on T cell recognition was seen. The results are discussed in light of the proposed domain structure of the class I proteins and the potential use of such molecules for further functional studies. PMID- 2985706 TI - Recombinant gamma-interferon induces changes in expression and shedding of antigens associated with normal human melanocytes, nevus cells, and primary and metastatic melanoma cells. AB - Recombinant gamma-interferon (rIFN-gamma) induced or augmented the expression of HLA-DR class II antigens on melanocytes isolated from newborn foreskin, from congenital, common acquired, and dysplastic nevi, and from primary and metastatic melanoma. The stimulatory effect of rIFN-gamma on HLA-DR antigen expression was suppressed by the addition of the phorbol ester TPA or its analog PDBu to the culture medium. Whereas rIFN-gamma did not significantly alter the expression of melanoma-associated, non-class II antigens on melanoma cells, there was a marked decrease in the expression of antigens associated with nevus cells. In addition, rIFN-gamma stimulated shedding of antigens. Increased antigen shedding was most apparent for an intracytoplasmic melanoma-associated protein of 80kd, followed by the ganglioside GD2 and by an alkali labile ganglioside. The simultaneous stimulation of class II antigen expression and shedding of melanoma-associated antigens as well as suppression of nevus-associated antigen expression could play an important role in the host immune response to premalignant and malignant melanocytic lesions. PMID- 2985707 TI - Optimization of conditions for in vitro antigenic stimulation of dissociated mouse spleen cells for the production of monoclonal antibodies against peptide hormones. AB - Factors affecting hybridoma yield following in vitro immunization have been investigated. Of critical importance for optimum yield was the batch of young calf serum used both in immunization cultures and in post-fusion cultures. Only 1 batch of serum was found to be suitable for the immunization step. The addition of horse serum to deficient young calf serum in pre-fusion cultures did not reconstitute the essential component(s). Addition of T cell, macrophage and bovine endothelial cell conditioned medium to the supportive batch of young calf serum in pre-fusion cultures did not increase the yield of hybridomas. For one antigen (insulin) the yield of hybrid cells was dependent on the concentration of antigen in immunization cultures. This was not the case, however, with the second antigen (ACTH) within the concentration range examined. The optimum spleen cells density was 1 X 10(7) cells/ml and the optimum culture period before fusion was 5 6 days. Although in vivo priming followed by in vitro boosting decreased the yield of hybrids the relative percentage of positive hybrids was slightly increased. The results suggest that in vivo priming is not essential since primary in vitro stimulation alone produced significant numbers of hybrids secreting specific antibody. PMID- 2985708 TI - Cytomegaloviral infection and disease. PMID- 2985709 TI - Trimethoprim resistance in multiple genera of Enterobacteriaceae at a U.S. hospital: spread of the type II dihydrofolate reductase gene by a single plasmid. AB - The percentage of clinical isolates of several species of Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae, resistant to trimethoprim (TMPR) has increased gradually at the Brigham and Women's Hospital (Boston) in recent years. Thirty-seven of 42 TMPR isolates from six species of gram-negative bacilli conjugally transferred TMP resistance to K12 E. coli. beta Lactam resistance cotransferred from 21 of the 37 donors, and sulfamethoxazole (SMZ) resistance cotransferred from five of the 37 donors. Plasmids that encoded TMP resistance either alone or with SMZ resistance had a molecular size of approximately 52.5 kilobases, with identical restriction endonuclease-generated "fingerprints." Plasmids encoding beta-lactam-mediated resistance (beta R) were approximately four kilobases larger and had fragment patterns that were identical for all of the TMPR/beta R plasmids tested and had many restriction endonuclease generated bands in common with TMPR plasmids. Radiolabeled dihydrofolate reductase (DHFR) probes identified the type II DHFR as the determinant of TMP resistance. In contrast with reports from Europe, TMP resistance in multiple species of Enterobacteriaceae was found to be spread in one hospital by a single, stable conjugative plasmid that has a wide host range and encodes the type II DHFR gene. PMID- 2985710 TI - Human enteric coronaviruses: antigenic relatedness to human coronavirus OC43 and possible etiologic role in viral gastroenteritis. AB - In a group of infants with acute nonbacterial gastroenteritis, examination of paired sera for antibody to human coronavirus (HCV) OC43 and neonatal calf diarrhea coronavirus showed a peculiar pattern of serological response, restricted only to HCV OC43 surface antigens, in a significantly higher proportion than among age-matched controls. In another group of infants and young children with acute nonbacterial gastroenteritis, fecal excretion of coronavirus like particles was detected by electron microscopy in 34 (16.3%) of 208 patients as compared with three (1.6%) of 182 controls (P less than .01). Two strains of human enteric coronavirus (HECV) were purified from stools of two patients, and immune sera were raised in mice and guinea pigs. Immune electron microscopy showed a two-way cross-reactivity between HECV and HCV OC43 when tested with immune sera and convalescent-phase sera from patients with infection due to HECV or HCV OC43. PMID- 2985711 TI - An outbreak of chancroid in Orange County, California: descriptive epidemiology and disease-control measures. AB - From May 1981 through February 1983, greater than 1,700 patients were examined for genital ulcers at the Orange County Special Diseases Clinic in Santa Ana, California. Of these patients, 923 had either confirmed chancroid or genital ulcers of unknown etiology. Haemophilus ducreyi was recovered from lesions or inguinal buboes of 271 patients. In the previous year, no cases of chancroid were reported in this community. Men accounted for 266 (98%) of the confirmed cases; 95% of the men were Hispanic, and at least 53% had had sexual contact with a prostitute. All five culture-positive women were prostitutes. Antimicrobial sensitivity testing showed resistance to both sulfamethoxazole and tetracycline but susceptibility to erythromycin and to the combination trimethoprim sulfamethoxazole. Treatment of patients with chancroid, their sex partners, and temporarily incarcerated prostitutes contributed to the successful control of this outbreak. PMID- 2985714 TI - Cofactors in disease: Epstein-Barr virus, oncogenes, and Burkitt's lymphoma. PMID- 2985713 TI - Acquisition of cytomegaloviral infections in families with young children: a serological study. AB - Acquisition of cytomegalovirus (CMV) infection in families with young children was investigated with serial serological tests for antibody to CMV by using ELISA and anti-complement immunoflourescence. Members of 68 Houston families were studied for a mean of 3.5 years. Seroconversion occurred in one or more members of 37 (53%) of 68 families studied. The mean annual seroconversion rate was 10% for fathers, 10.6% for mothers, and 10.3% for children. The attack rate for susceptible family members (including the index case) was 54%. The index case was the child in 10 instances, the father in 2, and the mother in 2. The index case could not be identified in 23 families. This Houston study indicates that the family with young children is a high-risk setting for CMV infection. This is an important concern regarding risks for mothers in their childbearing and child rearing years. PMID- 2985712 TI - IgG subclass antibodies to herpes simplex virus. AB - Several mouse monoclonal antibodies specific for human IgG1, IgG2, IgG3, and IgG4 were evaluated by enzyme-linked immunosorbent assay to detect human IgG subclass antibodies to herpes simplex virus (HSV) antigens. The variable results with different monoclonal antibodies point to the need for well-characterized reagents in the study of antibody responses to infectious agents. The 204 sera tested were obtained from 157 patients with various forms of clinically manifest HSV infections and from several controls. IgG1 antibodies were demonstrated in almost all HSV-infected subjects and were the first antibodies to appear in primary genital infections. IgG2, IgG3, and IgG4 antibodies were detected in acute-phase sera, most often in patients with recurrent genital herpes but in none of those with primary infections. IgG4 antibodies occurred significantly more frequently in sera from men than in those from women with recurrent genital infections. PMID- 2985715 TI - An inhibitor of interferon action: II. Biological properties of the IFN-gamma associated inhibitor of interferon action. AB - Previously an inhibitor of interferon action had been isolated from mitogen stimulated mouse spleen cells. This inhibitor was associated with the IFN-gamma molecule and might possibly represent an altered IFN-gamma molecule which can no longer induce an effective antiviral state. This report further investigates the biological properties of this inhibitor. Inhibitor activity is independent of the virus used in the interferon assay. Inhibitor activity has also been found to be species specific. Mouse inhibitor does not inhibit the antiviral activity of either human IFN-gamma or human IFN-beta. However, inhibitor production is not limited to the mouse system. Inhibitor is also present in preparations of human IFN-gamma. The inhibitor does not appear to directly compete with IFN-gamma for a specific cell surface receptor since inhibitor activity is independent of interferon concentration. The presence of inhibitor allows a unique, albeit reduced level of antiviral protection to develop. Increasing concentrations of interferon do not increase the level of antiviral protection allowed by the inhibitor. This inhibitor of interferon action may represent a natural mechanism whereby IFN-gamma-induced effects are regulated in vivo. PMID- 2985716 TI - Ameliorating effect of IFN-beta and anti-IFN-beta on coxsackievirus B3-induced myocarditis in mice. AB - A significant reduction in the number of virus-induced myocardial lesions was effected by administration of murine interferon beta (IFN-beta) or polyriboinosinic: polyribocytidylic acid copolymer (pI:pC) at -24, 0, or 24 h but not 72 h postinoculation (p.i.) of coxsackie-virus B3 (CVB3) to adolescent CD-1 mice. Inoculation of interferon at any of the four times did not reduce virus titers in heart tissues at three or seven days p.i., but inoculation of pI:pC at 24, 0, or 24 h p.i. significantly reduced virus titers. Administration of anti murine IFN-beta at 72 h p.i. significantly reduced myocarditic lesion numbers. The results suggest that there are two identifiable times after CVB3 inoculation in which interferon may play a role in CVB3-induced myocarditis: a very early time (+/- 24 h of virus entry) in which the presence of interferon is beneficial to the animal and a later time (72 h p.i. of virus) in which absence of interferon is beneficial to the animal. In vitro studies on the effects of IFN beta or anti-IFN-beta antiserum on replication of CVB3 in permissive primary cultures of murine neonatal skin fibroblasts show that this virus is sensitive to the antiviral action of interferon which is produced in infected cultures. PMID- 2985717 TI - Induction of human interferon gamma in nylon-column nonadherent human lymphocyte cells by heat-treated poly I:poly C. AB - Human peripheral blood mononuclear leukocytes (PBML) were separated by Ficoll hypaque gradient from human peripheral whole blood cells. The PBML were further separated by nylon-column. When nylon-column nonadherent cells (NNA cells) were induced by poly I:poly C which was prepared by heat-treatment, high levels of acid- and heat-labile interferon (IFN) were obtained. The IFN was not neutralized with human IFN-alpha or -beta anti-serum, but was completely neutralized with human IFN-gamma anti-serum, indicating that the IFN was human IFN-gamma. PMID- 2985718 TI - [Binding of human chorionic gonadotropin with insufficient terminal sugar to peanut agglutinin]. AB - We have studied the binding of human chorionic gonadotropin (hCG) in sera and urine of normal pregnant women and patients with trophoblastic diseases to the lectin from peanut (Arachis hypogaea, PNA). As the basic experiments, purified standard hCG and its alpha and beta subunits were examined for binding to a PNA Sepharose column. Both standard hCG and hCG-beta adsorbed to the PNA-Sepharose only when they were treated with neuraminidase, whereas hCG-alpha was still not adsorbed even after treatment with neuraminidase. The major proportions of hCG in sera and urine from normal pregnancy and hydatidiform mole were not adsorbed to the PNA-Sepharose column. On the other hand, the proportions of PNA binding fraction in sera and urine from choriocarcinoma were significantly elevated in 4 of 6 cases. The PNA binding fraction was significantly higher in serum than in urine of each individual. The PNA binding fractions were extracted and purified from urine of 2 patients with choriocarcinoma, and were quantitatively found to be asialo-hCG. These results suggest that PNA-Sepharose affinity chromatography is an effective tool to use in detecting incompletely sialylated hCG and hCG-beta and may be a useful index for the clinical diagnosis of choriocarcinoma. PMID- 2985719 TI - [Regional registration study of trophoblastic disease in Japan]. PMID- 2985720 TI - [Ovarian tumor and steroid hormone receptors]. PMID- 2985721 TI - Kinetics of inflammatory and fibrotic pulmonary changes in a murine model of silicosis. AB - A murine model of experimental silicosis has been developed after the intratracheal injection of alpha-quartz crystals. Pulmonary inflammation was monitored by increases in wet lung weight and cell number and protein content of the lung lavage fluid; fibrosis was assessed by measuring increases in hydroxyproline content of the lungs. Acute pulmonary cellular inflammation occurred between weeks 1 and 2, followed by a chronic inflammatory response at week 12. Lung hydroxyproline content, an indication of collagen deposition, was initiated as early as 1 week after silica injection and continued to increase steadily over time. The inflammatory and fibrotic changes induced by silica appeared to be a specific effect of the injection of this toxic particulate and not the result of the introduction of a foreign body, because mice injected with silica crystals were found to have significantly greater increases in acute cellular inflammation and chronic collagen deposition than did mice injected with latex beads. A possible role for the immune system in modulating silica-induced damage was suggested by the variability in response of six different strains of mice (C3H/He, CBA/J, Balb/c, DBA/2, C57BL/6, C57BL/10), which differed at specific genetic loci. Both strains with high (DBA/2) and low (C3H/He) response demonstrated similar patterns of inflammation and fibrosis over a period of 12 weeks. This model demonstrates great potential in future studies for elucidating the role of the immune system in the development of pulmonary inflammation and fibrosis induced by toxic inorganic particulates. PMID- 2985723 TI - Improved technic for detecting intraerythrocytic inclusion bodies in alpha thalassemia trait. PMID- 2985722 TI - Synthesis and structure of 26 (or 27)-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24S,25 xi-pentol isolated from the urine and feces of a patient with sitosterolemia and xanthomatosis. AB - The urine and feces of a patient with the rare inherited lipid storage disease, sitosterolemia and xanthomatosis, were analyzed. Substantial quantities of C26 bile alcohol, 26 (or 27)-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24S,25 xi pentol along with 5 beta-cholestane-3 alpha,7 alpha,12 alpha,24-tetrol, 5 beta cholestane-3 alpha,7 alpha,12 alpha,25-tetrol, 5 beta-cholestane-3 alpha,7 alpha,12 alpha,24R,25-pentol, and 5 beta-cholestane-3 alpha,7 alpha,12 alpha,25,26-pentol were found. The structure of the C26-bile alcohol was confirmed by direct comparison (gas-liquid chromatography-mass spectrometry and thin-layer chromatography) with a standard sample synthesized from cholic acid. The configurational assignment at C-24 was determined by lanthanide-induced circular dichroism Cotton effect measurements. The increased excretion of these C26- and C27-bile alcohols suggests an abnormality of bile acid biosynthesis in this disease. PMID- 2985724 TI - Stimulation of androgen and oestrogen concentrations in plasma of cows after administration of a synthetic glucocorticoid (flumethasone) at the end of gestation. AB - Administration of flumethasone (3.5 mg i.m.) to six cows on day 260 of pregnancy induced parturition in only one animal. In the other five cows circulating concentrations of epitestosterone and conjugated oestrogens increased to reach maximum values (epitestosterone, 13.73 +/- 2.81 nmol/l; conjugated oestrogens, 33.59 +/- 6.87 nmol/l) 2-3 days after treatment. Concentrations of these steroids were raised as long as the synthetic glucocorticoid was present in the circulation (as judged by depression of cortisol concentrations). After clearance of the drug, concentrations of these steroids declined to values present in the control group. Concentrations of unconjugated oestrogens were only slightly raised after flumethasone. In contrast to the effect observed after administration on day 260, treatment on day 270 induced parturition in four of six treated animals. In these cows, mean circulating concentrations of epitestosterone, unconjugated and conjugated oestrogens increased to 9.50 +/- 2.96, 9.62 +/- 1.48 and 36.51 +/- 4.8 nmol/l respectively to reach concentrations observed in the control group at parturition. After parturition the concentrations of epitestosterone and oestrogens declined rapidly in all groups. PMID- 2985725 TI - A black background facilitates the response to stress in teleosts. AB - This work examines the difference in responsiveness to stress which characterizes fish adapted to white and black backgrounds. Trout were maintained in black or white tanks for 2 weeks and then subjected to intermittent intense or moderate noise stress for periods between 1 h and 5 days, or to the stress of being injected daily with a large volume of liquid for 3 days. Plasma cortisol concentrations increased more readily and to a greater extent in fish from black tanks in response to moderate stress or brief intense stress. Dexamethasone suppressed the stress-induced rise of cortisol in white-adapted fish but was only partially effective in trout from black backgrounds. These differences in plasma cortisol between black- and white-adapted fish can be related to the different titres of plasma ACTH, apparently derived from the pars distalis. Removal of the neurointermediate lobe (NIL) from black-adapted eels markedly depressed the normal rise in plasma cortisol elicited by noise stress. It is suggested that products from the NIL may modulate the stress response of the hypothalamo pituitary axis although other routes through which background colour could affect the pituitary responsiveness to stress are also considered. In several cases, stress also enhanced the secretion of MSH from the NIL. PMID- 2985726 TI - Mechanisms of release of prolactin from fowl anterior pituitary glands incubated in vitro: effects of calcium and cyclic adenosine monophosphate. AB - Fowl anterior pituitary glands were bisected and each half was pretreated in either Medium 199 or medium containing EGTA to deplete endogenous calcium (Ca2+) stores, after which they were incubated in Medium 199, or Ca2+-free medium, containing prolactin release-stimulating agents and verapamil, a Ca2+ channel blocker. High K+ concentrations, hypothalamic extract, synthetic thyrotrophin releasing hormone (TRH) and dibutyryl cyclic AMP (dbcAMP) all stimulated release of prolactin from control (non EGTA-treated) hemianterior pituitary glands. The effects of TRH and dbcAMP were not additive, but the response to submaximal concentrations of TRH was augmented by theophylline, a phosphodiesterase inhibitor. Reduction of Ca2+ availability with EGTA or verapamil reduced basal release of prolactin, prevented the prolactin-stimulating effects of high K+ concentrations and TRH, and markedly attenuated responses to hypothalamic extract and dbcAMP, EGTA being more effective than verapamil. Increasing the Ca2+ concentration of the medium did not augment basal or stimulated release of prolactin. These results suggest that both Ca2+ and cyclic AMP may act as intracellular mediators in the release of prolactin. Both basal and stimulated release of prolactin depend upon the presence of Ca2+. Although influx from the medium may be the major source of Ca2+, endogenous stores of Ca2+, perhaps mobilized by dbcAMP, may be able to maintain some release of prolactin. The prolactin-stimulating effects of TRH may be mediated by cyclic AMP. PMID- 2985727 TI - Oestrogenic regulation of testicular androgen production during development in the rat. AB - The influence of age on the sensitivity of the testis to oestrogens was investigated. Intact male rats at 10, 25, 40 and 53 days of age were injected s.c. with vehicle, 5 or 50 micrograms oestradiol or diethylstilboestrol (DES)/100 g body wt twice daily for 2 days; the animals were killed 12 h after the last injection. Subsequently, the concentrations of testicular androgens and serum LH, prolactin, testosterone and androstanediol (5 alpha-androstane-3 alpha, 17 beta diol) were measured. Testicular androgen production was determined in vitro in the presence or absence of human chorionic gonadotrophin (hCG) or dibutyryl cyclic AMP (dbcAMP). Androgens in the serum and testes displayed an age-related alternating pattern with androstanediol being the major androgen produced at 27 days of age. As a result of oestrogen treatment, serum LH concentrations were decreased while serum prolactin was increased. Serum testosterone was decreased by 36-55% in the 12-day-old group and further reduced by 95% of control values by day 55; serum androstanediol was less sensitive to oestrogen suppression. Testicular concentrations of both testosterone and androstanediol exhibited a marked reduction in 12-day-old animals as a result of oestrogen administration. These values were reduced by 85-95% at day 27 and remained suppressed even at 55 days. Basal production of testosterone was unaffected by oestrogen treatment in 12- and 27-day-old animals but was markedly decreased by day 42. Significant suppression of basal production of androstanediol was observed as early as day 12.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985728 TI - The mammalian adrenal circulation and the relationship between adrenal blood flow and steroidogenesis. PMID- 2985729 TI - Heart-specific autoantibodies following murine coxsackievirus B3 myocarditis. AB - The sera from A.SW/SnJ mice infected with Coxsackievirus B3 (CB3) were tested on normal mouse tissue by indirect immunofluorescence. Heart-reactive antibodies were found. Absorption studies with organ extracts showed some of these autoantibodies to be heart-specific. Additional antibodies were crossreactive with skeletal muscle and kidney. These findings suggest a role for autoimmunity in the pathogenesis of murine CB3-induced myocarditis. This study establishes an animal model for the study of the humoral autoimmune response in human viral myocarditis. PMID- 2985730 TI - Interleukin 2 inhibits in vitro growth of human T cell lines carrying retrovirus. AB - Four human T cell lines, TL-Mor, TL-Su, TL-TerI, and TL-OmI, carrying human T cell leukemia virus (HTLV), were established previously. TL-Mor, TL-Su, and TL TerI were derived from interleukin 2 (IL-2)-dependent parental cell lines cloned from peripheral blood leukocytes (PBL) of three healthy HTLV carriers, while TL OmI was directly established from PBL of a patient with adult T cell leukemia. These four TL cell lines grow autonomously without IL-2. When they were cultured in the presence of IL-2, their growth was inhibited after a few days. This growth inhibition depended on the dose of IL-2, and the effective dose significantly promoted growth of their parental IL-2-dependent cell lines. The growth inhibition is demonstrated to be due to specific binding of IL-2 to receptors on the TL cells. PMID- 2985731 TI - Rat lymphoid cell lines producing human T cell leukemia virus. II. Constitutive expression of rat interleukin 2 receptor. AB - Three rat lymphoid cell lines (TARS-1, TARL-2, and TART-1) (12) transformed by human T cell leukemia/lymphoma virus I (HTLV-I) had rearrangement of the beta chain gene of the T cell antigen receptor, and had integrated proviral DNA from HTLV-I in their genomes. As is the case with adult T cell leukemia (ATL)-derived human T cell lines transformed by HTLV-I, these rat cell lines unequivocally expressed interleukin 2 (IL-2) receptor, as determined by radiolabeled IL-2 binding. By Scatchard plot analysis, one of the cell lines, TART-1, proved to have high affinity receptors (Ka = 1.3 X 10(11)/M and 8.8 X 10(9)/M). Rat IL-2 receptor, not human IL-2 receptor, was expressed on HTLV+ rat cell lines, as demonstrated by the fact that they expressed antigens reactive with monoclonal antibodies (ART-18) against rat IL-2 receptor, but not with anti-Tac antibodies. The collective evidence indicates that the endogenous IL-2 receptor gene is activated in human and rat lymphoid cell lines with HTLV-I production. The mechanism of abnormal IL-2 receptor expression in HTLV infection is discussed. PMID- 2985733 TI - Effect of forskolin on the spontaneous maturation and cyclic AMP content of hamster oocyte-cumulus complexes. AB - Forskolin, a reversible stimulator of the catalytic subunit of adenylate cyclase, has been used to determine: whether an increase in hamster cumulus cell cyclic adenosine monophosphate (cAMP) results in an elevation of intraoocyte cAMP and an accompanying increase in the maintenance of meiotic arrest (%GV where GV is germinal vesicle) when heterologous coupling is maintained, whether the hamster oolemma possesses the catalytic subunit of adenylate cyclase in an amount adequate to stimulate sufficient cAMP synthesis to maintain arrest, and whether release from meiotic arrest is accompanied by a decrease in the content of intraoocyte cAMP. Intracellular cAMP was determined by RIA, functional metabolic coupling was assessed by determination of the fraction of radiolabeled uridine marker transferred from the cumulus mass to the oocyte, and meiotic stage was determined cytogenetically. While the %GV of both cumulus-enclosed (intact) and cumulus-free (denuded) oocytes was dose-dependent upon forskolin, that of intact oocytes was much more sensitive to the drug (intact: ID50 3.4 microM; denuded: ID50 65.0 microM, where ID50 is the dose of forskolin that inhibits the maturation of 50% of cultured oocytes). Forskolin stimulated a significant, dose dependent increase in the amount of cAMP within the cumulus mass [(r) = 0.789, P less than 0.001)], the intact oocyte [(r) = 0.715, P less than 0.001], and the denuded oocyte [(r) = 0.673, P less than 0.01)]. The cAMP content of intact oocytes was significantly greater than that of denuded oocytes above 6.25 microM forskolin (25 microM forskolin: 9.28 +/- 1.01 vs. 3.98 +/- 0.15 fmol cAMP, intact and denuded oocytes, respectively; P less than 0.001, paired t test). A highly significant positive correlation was established between the amount of cAMP in groups of cumulus masses and that in the corresponding enclosed oocytes [(r) = 0.635, P less than 0.001]. The enhanced sensitivity of meiotic arrest in intact, as compared to denuded, oocytes was due to the presence of adherent cumulus cells but was not attributable to a significant increase in the cAMP content of intact oocytes (at 6.25 microM forskolin; %GV intact = 73.0 +/- 10.7, denuded = 20.3 +/- 7.4; fmol cAMP intact = 5.02 +/- 1.50; denuded = 4.63 +/- 0.81). The arresting action of forskolin on intact oocytes was transient and fully reversible, but release from arrest was not accompanied by a decrease in either intraoocyte cAMP or heterologous metabolic coupling.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985732 TI - Lack of renal effects of DOCA, ACTH, spironolactone, and angiotensin II in Squalus acanthias. AB - Angiotensin II was infused intravenously in spiny dogfish sharks (Squalus acanthias). There were no significant effects on arterial blood pressure, glomerular filtration rate, urine flow, or Na excretion either in comparison with pre- and postinfusion values or in comparison with values measured in a control group of fish given elasmobranch saline intravenously. In other dogfish, glomerular filtration rate, urine flow, and Na and K excretory rates were measured for 3 days following implantation of desoxycorticosterone (DOCA), adrenocorticotropin (ACTH), or spironolactone; a control group was given no drug. There were no significant differences between these four groups of fish with respect to any of the measured parameters. These results suggest that the dogfish kidney is not a target organ for several substances known to affect renal function, either directly or indirectly, in other animals. PMID- 2985734 TI - Intracellular pH-regulating mechanism of the squid axon. Relation between the external Na+ and HCO-3 dependences. AB - The intracellular pH-regulating mechanism of the squid axon was examined for its dependence on the concentrations of external Na+ and HCO3-, always at an external pH (pHo) of 8.0. Axons having an initial intracellular pH (pHi) of approximately 7.4 were internally dialyzed with a solution of pH 6.5 that contained 400 mM Cl- and no Na+. After pHi had fallen to approximately 6.6, dialysis was halted, thereby returning control of pHi to the axon. With external Na+ and HCO-3 present, intracellular pH (pHi) increased because of the activity of the pHi regulating system. The acid extrusion rate (i.e., equivalent efflux of H+, JH) is the product of the pHi recovery rate, intracellular buffering power, and the volume-to-surface ratio. The [HCO3-]o dependence of JH was examined at three fixed levels of [Na+]o: 425, 212, and 106 mM. In all three cases, the apparent Jmax was approximately 19 pmol X cm-2 X s-1. However, the apparent Km (HCO3-) was approximately inversely proportional to [Na+]o, rising from 2.6 to 5.4 to 9.7 mM as [Na+]o was lowered from 425 to 212 to 106 mM, respectively. The [Na+]o dependence of JH was similarly examined at three fixed levels of [HCO3-]o: 12, 6, and 3 mM. The Jmax values did not vary significantly from those in the first series of experiments. The apparent Km (Na+), however, was approximately inversely related to [HCO3-]o, rising from 71 to 174 to 261 mM as [HCO3-]o was lowered from 12 to 6 to 3 mM, respectively. These results agree with the predictions of the ion-pair model of acid extrusion, which has external Na+ and CO3= combining to form the ion pair NaCO3-, which then exchanges for internal Cl . When the JH data are replotted as a function of [NaCO3-]o, data from all six groups of experiments fall along the same Michaelis-Menten curve, with an apparent Km (NaCO3-) of 80 microM. The ordered and random binding of Na+ and CO3= cannot be ruled out as possible models, but are restricted in allowable combinations of rate constants. PMID- 2985735 TI - Cyclic AMP inhibits Na+/H+ exchange at the apical membrane of Necturus gallbladder epithelium. AB - The effects of elevating intracellular cAMP levels on Na+ transport across the apical membrane of Necturus gallbladder epithelium were studied by intracellular and extracellular microelectrode techniques. Intracellular cAMP was raised by serosal addition of the phosphodiesterase inhibitor theophylline (3 mM) or mucosal addition of either 8-Br-cAMP (1 mM) or the adenylate cyclase activator forskolin (10 microM). During elevation of intracellular cAMP, intracellular Na+ activity (alpha Nai) and intracellular pH (pHi) decreased significantly. In addition, acidification of the mucosal solution, which contained either 100 or 10 mM Na+, was inhibited by approximately 50%. The inhibition was independent of the presence of Cl- in the bathing media. The rates of change of alpha Nai upon rapid alterations of mucosal [Na+] from 100 to 10 mM and from 10 to 100 mM were both decreased, and the rate of pHi recovery upon acid loading was also reduced by elevated cAMP levels. Inhibition was approximately 50% for all of these processes. These results indicate that cAMP inhibits apical membrane Na+/H+ exchange. The results of measurements of pHi recovery at 10 and 100 mM mucosal [Na+] and a kinetic analysis of recovery as a function of pHi suggest that the main or sole mechanism of the inhibitory effect of cAMP is a reduction in the maximal rate of acid extrusion. In conjunction with the increase in apical membrane electrodiffusional Cl- permeability, produced by cAMP, which causes a decrease in net Cl- entry (Petersen, K.-U., and L. Reuss, 1983, J. Gen. Physiol., 81:705), inhibition of Na+/H+ exchange contributes to the reduction of fluid absorption elicited by this agent. Similar mechanisms may account for the effects of cAMP in other epithelia with similar transport properties. It is also possible that inhibition of Na+/H+ exchange by cAMP plays a role in the regulation of pHi in other cell types. PMID- 2985736 TI - Norepinephrine as a possible transmitter involved in synaptic transmission in frog taste organs and Ca dependence of its release. AB - In order to explore the role of catecholamine and Ca2+ in the synaptic transmission from taste cells to sensory nerve terminals, the effects of various agents added to an artificial solution perfusing the lingual artery on the frog taste nerve responses were examined. The injection of reserpine or guanetidine, which are catecholamine-depleting agents, led to a great reduction of the frog taste nerve responses. The addition of catecholamines to the perfusing solution did not practically enhance the spontaneous impulse discharges, but did recover the response to all the taste stimuli examined. Norepinephrine was most effective and is the most likely candidate for the transmitter. The enhancement of the responses by norepinephrine was suppressed by desipramine, cocaine, or imipramine, which suggests that the enhancement was brought about by incorporation of norepinephrine into taste cells. In a previous paper (Nagahama, S., Y. Kobatake, and K. Kurihara, 1982. J. Gen. Physiol. 80:785), we showed that the responses to the stimuli of one group depended on Ca2+, cGMP, and cAMP added to the perfusing solution and those to the stimuli of another group did not depend on these agents. After the injection or addition of reserpine to the lingual artery, which probably modified injection or addition of reserpine to the lingual artery, which probably modified the permeability of the artery, the responses to the stimuli of the latter group also came to exhibit dependences on these agents, which indicates that the responses to all the taste stimuli have dependences on Ca2+, cGMP, and cAMP. PMID- 2985737 TI - Regulation of intracellular pH in human neutrophils. AB - The intracellular pH (pHi) of isolated human peripheral blood neutrophils was measured from the fluorescence of 6-carboxyfluorescein (6-CF) and from the equilibrium distribution of [14C]5,5-dimethyloxazolidine -2,4-dione (DMO). At an extracellular pH (pHo) of 7.40 in nominally CO2-free medium, the steady state pHi using either indicator was approximately 7.25. When pHo was suddenly raised from 7.40 to 8.40 in the nominal absence of CO2, pHi slowly rose by approximately 0.35 during the subsequent hour. A change of similar magnitude in the opposite direction occurred when pHo was reduced to 6.40. Both changes were reversible. Intrinsic intracellular buffering power, determined by using graded pulses of CO2 or NH4Cl, was approximately 50 mM/pH over the pHi range of 6.8-7.9. The course of pHi obtained from the distribution of DMO was followed during and after imposition of intracellular acid and alkaline loads. Intracellular acidification was brought about either by exposing cells to 18% CO2 or by prepulsing with 30 mM NH4Cl, while pHo was maintained at 7.40. In both instances, pHi (6.80 and 6.45, respectively) recovered toward the control value at rates of 0.029 and 0.134 pH/min. These rates were reduced by approximately 90% either by 1 mM amiloride or by replacement of extracellular Na with N-methyl-D-glucamine. Recovery was not affected by 1 mM SITS or by 40 mM alpha-cyano-4-hydroxycinnamate (CHC), which inhibits anion exchange in neutrophils. Therefore, recovery from acid loading is probably due to an exchange of internal H for external Na. Intracellular alkalinization was achieved by exposing the cells to 30 mM NH4Cl or by prepulsing with 18% CO2, both at a constant pHo 7.40. In both instances, pHi, which was 7.65 and 7.76, respectively, recovered to the control value. The recovery rates (0.033 and 0.077 pH/min, respectively) were reduced by 80-90% either by 40 mM CHC or by replacement of extracellular Cl with p-aminohippurate (PAH). SITS, amiloride, and ouabain (0.1 mM) were ineffective.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985738 TI - Bacteriophage P1 derivatives unaffected in their growth by a large inversion or by IS insertions at various locations. AB - Several plaque-forming phage P1 derivatives carrying DNA rearrangements associated with IS elements are described. They have IS1, IS3 and IS5 inserted in four distinct locations, all of which are non-essential regions for phage P1 propagation. One derivative carries a genome segment, inverted relative to the one in the P1 wild-type genome, between two inverted copies of IS1. The inverted DNA segment spans about 23 kb of the 90 kb long P1 genome and it includes the invertible C segment. This phage is as viable as an isomeric P1 which carries the relevant segment in its original orientation. These results are discussed with regard to the genome organization of phage P1. PMID- 2985739 TI - Molecular cloning of a pIP501 derivative yields a model replicon for the study of streptococcal conjugation. AB - We have previously constructed a derivative of the broad host range streptococcal plasmid pIP501, a conjugative plasmid designated pVA797, that confers chloramphenicol resistance and contains a unique EcoRI site in a non-essential region of the plasmid molecule. pVA797 (30.7 kb) when cloned in toto as an EcoRI fragment into the positive selection vector pOP203(A2+) gave a recombinant, pVA904 (37.7 kb), which was able to replicate in Escherichia coli and in streptococcal species. It can be phenotypically monitored in either genus by specific drug resistance markers (chloramphenicol resistance in streptococci, tetracycline resistance in E. coli). pVA904 segregates into E. coli minicells where it specifies the production of at least 13 polypeptides. Many of the polypeptides are missing in minicells containing a transfer-defective, deletion derivative of pVA904. pVA904 is an ideal model replicon for the study of streptococcal conjugation because it is a shuttle plasmid thus enabling manipulation using procedures established for E. coli. Specifically, it should be possible to define the genetic basis of streptococcal conjugation by coupling mutagenesis protocols and minicell protein analyses in E. coli with evaluation of transfer function in streptococci. PMID- 2985740 TI - Metronidazole radical anion generation in vivo in Trichomonas vaginalis: oxygen quenching is enhanced in a drug-resistant strain. AB - The nitro radical-anion of metronidazole has been detected in vivo in the sexually transmitted human parasite, Trichomonas vaginalis, under anaerobic conditions by electron spin resonance spectrometry. Exposure of organisms to oxygen decreased the intensity of the radical signal in both metronidazole sensitive ATCC strain 30001 and in the metronidazole-resistant strain 85. The sensitive strain still gave radical signals at partial pressures of oxygen (greater than 6 kPa) sufficient to remove all detectable radicals from the resistant strain. This evidence suggests that the resistant strain has defective oxygen scavenging system(s). PMID- 2985741 TI - Evolution of an R plasmid from a cryptic plasmid by transposition of two copies of Tn1 in Providencia stuartii. AB - Examination of a series of isolates of Providencia stuartii collected over an 18 month period from a chronic-care patient at Bristol Royal Infirmary revealed the emergence of resistance to carbenicillin. Resistance was mediated by a 47 kb plasmid which transferred by conjugation to a plasmid-free strain of P. stuartii but not to Escherichia coli. Carbenicillin-sensitive isolates were either plasmid free or contained a 36 kb cryptic plasmid. Restriction endonuclease mapping of this plasmid showed it to be closely related to 32 kb and 34 kb cryptic plasmids reported previously in P. stuartii from Bristol. Mapping of the R plasmid showed it to be derived from the 34 kb cryptic plasmid by transposition of two copies of Tn1. PMID- 2985742 TI - Three restriction endonucleases from Anabaena flos-aquae. AB - Three site-specific endonucleases, AflI, AflII and AflIII, have been partially purified from the cyanobacterium Anabaena flos-aquae CCAP 1403/13f. Their recognition and cleavage specificities have been determined to be: (formula; see text) AflII and AflIII are new specificities and may be useful in molecular cloning, as well as in the analysis of DNA. The distribution of type II restriction endonucleases in the cyanobacteria is briefly discussed. PMID- 2985743 TI - Impact of a case manager program on psychiatric aftercare. AB - Shifting the locus of aftercare planning from hospital to community can enhance continuity of care. The authors compared chronically mentally ill patients assessed and managed by community-based practitioners trained in psychiatric rehabilitation with patients whose discharge planning was arranged by inpatient staff members. They found significant differences between the two groups in aftercare needs identified, aftercare referrals made, and use of aftercare services. The authors conclude that this approach to psychiatric aftercare is superior to more traditional models if practitioners are carefully trained. PMID- 2985744 TI - Interactions between naloxone and GABA in the control of locomotor activity in the rat. AB - It was found that naloxone causes a small but significant reduction of motility. The GABAB agonist baclofen and the GABA transaminase inhibitor gamma-acetylenic GABA (GAG) also reduced locomotor activity. When a subeffective dose of baclofen was combined with naloxone 0.8 or 3.2 mg/kg, baclofen significantly inhibited motility beyond the inhibition caused by naloxone + saline. GAG, in a dose of 12.5 mg/kg, was also potentiated by naloxone, 3.2 mg/kg. The locomotion reducing effects of naloxone could be blocked by either picrotoxin or bicuculline. It is concluded that GABAergic mechanisms participate in the inhibition of locomotor activity provoked by naloxone. The possibility that this drug disinhibits GABAergic neurons is discussed. PMID- 2985745 TI - Ring neuron control of columellar motor neurons during egg-laying behavior in the pond snail. AB - Egg-laying in Lymnaea is characterized by the stereotyped egg-laying behavior (ELB), composed of foot contractions and shell movements. Egg-laying can be induced by a clean water stimulus, that triggers a discharge of the neuroendocrine caudo-dorsal cells (CDCs), which release the ovulation hormone into the blood. A part of the behavior is lost when egg-laying is triggered by hormone injection, indicating that during natural stimulus-induced or spontaneous egg-laying this part (the first phase) may be controlled by neuronal events in the CNS triggered by (a) factor(s) not released to the blood. The authors have identified an unpaired neuron, the ring neuron, that is excited during an in vitro afterdischarge of the CDCs, and which, by its numerous axonal branches in the pedal ganglia, modulates motorneurons of the columellar muscle, which controls shell movements. These motor-neurons, identified as such in reduced preparations by 1 for 1 muscle potentials and elements in the connecting nerve, all receive either excitatory or inhibitory input from the ring neuron, as well as from an unknown neuron which has common input of the ring neuron and the motorneurons. The action of the CDCs on the ring neuron cannot be mimicked by the ovulation hormone, and we therefore conclude that the first part of the ELB is probably caused by a nonhormonal local action of the CDCs on the ring neuron and possibly the common input neuron. PMID- 2985746 TI - Quantification of ultrastructural symmetry at molluscan chemical synapses. AB - Two criteria were developed to identify symmetrical synapses in the tentacle ganglion of the snail Achatina fulica. First, the concentration of electron lucent vesicles within 20 nm of one membrane had to be less than three times the concentration at the opposed membrane. Second, paramembranous densities had to be present in both cells, as unanimously judged by three independent observers. Nine synapses (27% of sample) satisfied both criteria. It is suggested that these synapses transmit chemically in two directions. PMID- 2985747 TI - Neuromuscular relationships in the abdomen of the Californian shore crab Pachygrapsus crassipes. AB - There are two pairs of muscles in each abdominal segment of the crab; one pair of flexors and one pair of extensors. In the early larval stages the muscles have short sarcomeres--a property of fast fibers--and high thin to thick filament ratios--a property of slow fibers. In the adult the abdominal muscles are intermediate and slow, since they have fibers with intermediate and long sarcomeres, high thin to thick filament ratios, low myofibrillar ATPase activity, and high NADH diaphorase activity. The different fiber types are regionally distributed within the flexor muscle. Microelectrode recordings from single flexor muscle fibers in the adult showed that most fibers are supplied by three excitatory motor axons, although some are supplied by as many as five efferents. One axon supplies all of the flexor muscle fibers in its own hemisegment, and the evoked junctional potentials exhibit depression. This feature together with the innervation patterns of the fibers are similar to those reported for the deep flexor muscles of crayfish and lobsters. Therefore, in the adult crab, the abdominal flexor muscles have some features in common with the slow superficial flexors of crayfish and other features in common with the fast deep flexor muscles. PMID- 2985748 TI - Sprouting and functional regeneration of an identified serotonergic neuron following axotomy. AB - An identified serotonergic neuron (C1) in the cerebral ganglion of Helisoma trivolvis sprouts following axotomy and rapidly (seven to eight days) regenerates to recover its regulation of feeding motor output from neurons of the buccal ganglia. The morphologies of normal and regenerated neurons C1 were compared. Intracellular injection of the fluorescent dye, Lucifer Yellow, into neuron C1 was compared with serotonin immunofluorescent staining of the cerebral and buccal ganglia. The two techniques revealed different and complimentary representations of the morphology of neuron C1. Lucifer Yellow provided optimal staining of the soma, major axon branches, and dendritic arborization. Immunocytochemical staining revealed terminal axon branches on distant targets and showed an extensive plexus of fine fibers in the sheaths of ganglia and nerve trunks. In addition to C1, serotonin-like immunoreactivity was localized in approximately 30 other neurons in each of the paired cerebral ganglia. Only cerebral neurons C1 had axons projecting to the buccal ganglia. No neuronal somata in the buccal ganglia displayed serotonin-like immunoreactivity. Observations of regenerating neurons C1 demonstrated: Actively growing neurites, both in situ and in cell culture, displayed serotonin-like immunoreactivity; severed distal axons of C1 retained serotonin-like immunoreactivity for up to 28 days; axotomized neurons C1 regenerated to restore functional control over the feeding motor program. PMID- 2985749 TI - Long-term facilitation and long-term adaptation at synapses of a crayfish phasic motoneuron. AB - Stimulation of the phasic (fast) motor axon of the isolated crayfish claw preparation at relatively low frequency (0.1 Hz) leads to depression of the excitatory junction potential (EJP) recorded from single muscle fibers. When the same stimulation is delivered following depression of the EJP at a higher frequency (5 Hz), a potentiated EJP appears, which is more resistant to low frequency depression. The potentiation appears to be analogous to "long-term facilitation" observed after stimulation of a tonic motor axon in crayfish and crabs. Long-term facilitation can be detected in preparations made from claws of animals in which the phasic motoneuron was stimulated at 5 Hz for 2 h in situ. This effect lasts for at least one day after one conditioning trial. Long-term facilitation is observed after stimulation of decentralized axons in situ, indicating that the change is attributable to local changes in terminal regions of the axon, and does not require the cell body. When electrodes are implanted in situ and the phasic motoneuron stimulated at 5 Hz for 2 h each day, synaptic depression becomes less pronounced and initial EJP amplitude becomes smaller over a period of several days. The latter changes, which adapt the neuron to a more tonic activity pattern, usually require several days for completion. Adaptation of fatigability occurs more rapidly than adaptation of initial EJP amplitude, and once established, remains for many days without further superimposed activity. Long-term adaptation does not occur in decentralized axons. Long-term facilitation and long-term adaptation are different responses of the neuron to enhanced activity. The former can occur in isolated or decentralized axons and leads to enhancement of EJP amplitude for a period of several hours to at least one day after a single episode of conditioning. The latter requires more time to be established, and leads to reduction of initial EJP amplitude and to lessened fatigability which persists for many days. PMID- 2985750 TI - Benzodiazepine receptor and thyroid hormones: in vivo and in vitro modulation. AB - In rats rendered hyperthyroid by chronic treatment with L-triiodothyronine (T3) hormone there was a 21 and 27% decrease, respectively, in the number of binding sites for [3H]flunitrazepam ([3H]FNZ) and [3H]ethyl-beta-carboline-3-carboxylate ([3H]beta-CCE) without changes in affinity for the two ligands. Two weeks after thyroidectomy there was a 44% increase in [3H]FNZ sites and a 17% increase in [3H]beta-CCE binding sites. In vitro we found that T3 produces a decrease in Bmax and an increase in KD, both changes being characteristic of a mixed type of inhibition. Thyroid status dramatically affected the Ki of T3 in displacing [3H]FNZ from sites on isolated membranes of the cerebral cortex: in hypothyroid rats the Ki value was 0.9 microM, whereas in hyperthyroid rats, it was 83 microM, a 92-fold difference. In control rats, the Ki was 11 microM. These findings are discussed in relation to a possible modulation of benzodiazepine receptors by thyroid hormones. PMID- 2985752 TI - Glycemia, ketonemia, and brain enzymes of ketone body utilization in suckling and adult rats undernourished from intrauterine life. AB - The effect of undernutrition from the 16th day of pregnancy up to 70th day of life on blood glucose and ketone bodies and on several brain mitochondrial enzymes related to energy metabolism or biosynthetic function was investigated. Undernutrition in perinatal period was established by means of a food restriction to pregnant rats and, later, to the lactating mother; undernourished postweaned rats received half the diet consumed by the controls. Body and brain weight from undernourished rats was less than controls throughout the entire period studied. Glycemia and ketonemia were also always lower than controls. Cytochrome c oxidase, citrate synthase, 3-hydroxybutyrate dehydrogenase, 3-oxoacid coenzyme A transferase, and acetoacetyl-coenzyme A thiolase activities during the suckling period were in most stages lower than controls; subsequently, activities in undernourished rats reached or surpassed the control values. These results could explain the "catch up" phenomenon in several ultrastructural parameters found by other authors in undernourished postweaned rats. PMID- 2985751 TI - Cyclic AMP concentrations in rat neocortex and hippocampus during and following incomplete ischemia: effects of central noradrenergic neurons, prostaglandins, and adenosine. AB - The concentrations of cyclic AMP, noradrenaline, glycogen, glucose, lactate, pyruvate, labile phosphate compounds, and free fatty acids were investigated in the rat neocortex and hippocampus during and following cerebral ischemia. An incomplete ischemia of 5 and 15 min duration was induced by bilateral carotid clamping combined with hypotension. The postischemic events were studied after 5, 15, and 60 min of recirculation. Five minutes of ischemia did not significantly alter the neocortical or hippocampal concentrations of cyclic AMP. After 15 min of ischemia the neocortical levels decreased significantly below control values. In the recirculation period following ischemia a significant elevation of the cyclic AMP concentrations was observed. Following 5 min of recirculation after 5 min of ischemia the levels increased from 2.53 +/- 0.21 nmol X g-1 to 5.18 +/- 0.09 nmol X g-1 in the neocortex and from 2.14 +/- 0.16 nmol X g-1 to 3.52 +/- 0.35 nmol X g-1 in the hippocampus. Five minutes of recirculation following 15 min of ischemia led to a significant increase in the levels of cyclic AMP, to 12.86 +/- 1.43 nmol X g-1 in the neocortex to 5.58 +/- 0.57 nmol X g-1 in the hippocampus. With longer recirculation periods the cyclic AMP levels progressively decreased and were similar to control values after 60 min. Depletion of cortical noradrenaline by at least 95% was performed by injections of 6-hydroxydopamine into the ascending axon bundles from the locus ceruleus. The lesion did not significantly change the ischemic or post-ischemic neocortical and hippocampal levels of cyclic AMP, glycogen, or free fatty acids including arachidonic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985753 TI - Pre- versus postsynaptic localization of benzodiazepine and beta-carboline binding sites. AB - [3H]Flunitrazepam (FNP) and [3H]methyl beta-carboline-3-carboxylate (MCC) binding was examined in soluble and particulate fractions from membranes solubilized with Triton X-100 or in subfractions of synaptosomal membranes obtained by a physical separation technique. Results using both methods demonstrate that benzodiazepine and beta-carboline sites reside on both pre- and postsynaptic membranes. Further, subfractionation experiments indicate that the binding sites for both ligands are unequally distributed within the synapse and among brain regions. For example, in cerebral cortical presynaptic membranes there are twice as many FNP as MCC sites whereas in postsynaptic membranes this ratio is reversed. The number of FNP and MCC sites are equal in the presynaptic fraction from cerebellum. The postsynaptic membranes derived from cerebellum have three times the number of FNP compared to MCC sites. In hippocampus this ratio varies between 1.5 and 2.8 in each subfraction. These results support the idea that benzodiazepine and beta carboline binding sites represent different recognition sites. PMID- 2985754 TI - Fe2+-induced lysis and lipid peroxidation of chromaffin granules. AB - Chromaffin granules, the catecholaminergic storage granules from adrenal chromaffin cells, lysed in 10(-9)-10(-7) M Fe2+. Lysis was accompanied by the production of malondialdehyde which results from lipid peroxidation. Both chromaffin granule lysis and malondialdehyde production were inhibited by the free radical trapping agent butylated hydroxytoluene but not by catalase and/or superoxide dismutase. The results suggest that lysis resulted from a direct transfer of electrons from Fe2+ to a component of the chromaffin granule membrane without the participation of either superoxide or hydrogen peroxide and may have resulted from lipid peroxidation. In some experiments, ascorbate alone induced chromaffin granule lysis which was inhibited by EDTA, EGTA, or deferoxamine. The lysis was probably caused by trace amounts of reducible polyvalent cation. Lysis sometimes occurred when Ca2+ was added with EGTA (10 microM free Ca2+ concentration) and was consistently observed together with malondialdehyde production in the presence of Ca2+, EGTA, and 10 microM Fe2+ (total concentration). The apparent Ca2+ dependency for chromaffin granule lysis and malondialdehyde production was probably caused by a trace reducible polyvalent ion displaced by Ca2+ from EGTA and not by a Ca2+-dependent reaction involving the chromaffin granule. PMID- 2985755 TI - Identification of endogenous calmodulin-dependent kinase and calmodulin-binding proteins in cold-stable microtubule preparations from rat brain. AB - Calmodulin-dependent kinase activity was investigated in cold-stable microtubule fractions. Calmodulin-dependent kinase activity was enriched approximately 20 fold over cytosol in cold-stable microtubule preparations. Calmodulin-dependent kinase activity in cold-stable microtubule preparations phosphorylated microtubule-associated protein-2, alpha- and beta-tubulin, an 80,000-dalton doublet, and several minor phosphoproteins. The endogenous calmodulin-dependent kinase in cold-stable microtubule fractions was identical to a previously purified calmodulin-dependent kinase from rat brain by several criteria including (1) subunit molecular weights, (2) subunit isoelectric points, (3) calmodulin binding properties, (4) subunit autophosphorylation, (5) calmodulin-binding subunit composition on high-resolution sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (6) isolation of kinase on calmodulin affinity resin, (7) kinetic parameters, (8) phosphoamino acid phosphorylation sites on beta-tubulin, and (9) phosphopeptide mapping. Endogenous cold-stable calmodulin-dependent kinase activity was isolated from the microtubule fraction by calmodulin affinity resin column chromatography and specifically eluted with EGTA. This kinase fraction contained the calmodulin-binding, autophosphorylating rho and sigma subunits of the previously purified kinase. The rho and sigma subunits of this kinase represented the major calmodulin-binding proteins in the cold-stable microtubule fractions as assessed by denaturing and non-denaturing procedures. These results indicate that calmodulin-dependent kinase is a major calmodulin binding enzyme system in cold-stable microtubule fractions and may play an important role in mediating some of the effects of calcium on microtubule and cytoskeletal dynamics. PMID- 2985756 TI - Nerve growth factor affects cyclic AMP metabolism, but not by directly stimulating adenylate cyclase activity. AB - Published experiments both support and contradict the hypothesis that nerve growth factor (NGF) can regulate adenylate cyclase activity. Using a sensitive assay that measures the conversion of [2-3H]adenine to [3H]cyclic AMP, we have shown that NGF alone cannot measurably stimulate cyclic AMP production, whereas the adenosine analog phenylisopropyladenosine (PIA) stimulates adenylate cyclase 20-fold over basal activity. NGF potentiates the capacity of both PIA and cholera toxin to stimulate cyclic AMP accumulation at all concentrations tested. This potentiation occurs at the earliest measurable times and does not require RNA synthesis. Therefore, we conclude that cyclase activation alone does not account for the effect of NGF on cyclic AMP accumulation and we discuss possible mechanisms. PMID- 2985757 TI - Differences in binding properties of mu and delta opioid receptor subtypes from rat brain: kinetic analysis and effects of ions and nucleotides. AB - Differences in binding properties of mu and delta opioid receptors were investigated using DAGO (Tyr-D-Ala-Gly-MePhe-Gly-ol) and DTLET (Tyr-D-Thr-Gly-Phe Leu-Thr), which occur, respectively, as the most selective mu and delta radioligands available. At high concentration, each agonist is able to interact with its nonspecific sites. Competition experiments indicated that a two-site competitive model was adequate to explain the interactions of DAGO and DTLET with [3H]DTLET and [3H]DAGO binding sites, respectively. The weak cross-reactivity (congruent to 10%) of DTLET for mu sites was taken into account in these experiments. On the other hand, DAGO and DTLET exhibit differential binding kinetics. Thus, at 35 degrees C, the lifetime of DTLET within its receptor site is about 14 times longer than that of the mu agonist. Sodium and manganese ions decrease the maximal number of high affinity mu and delta sites, but the sensitivity of mu receptors is three times higher towards Na+ and 20-fold higher towards Mn2+ than that of delta receptors. GTP reduces similarly the mu and delta binding whereas only the DAGO binding was modified by the nonhydrolyzable analogue guanylylimidodiphosphate [GMP-P(NH)P]. However, in the presence of Na+ ions, GMP-P(NH)P inhibits the DTLET binding in a concentration-dependent manner. The effects of Na+ and GMP-P(NH)P could be explained by a sequential transformation of delta receptors to low-affinity states. This model predicts that Na+, by lowering the affinity of a fraction of sites, produces a decrease in the maximal number of high-affinity delta receptors and that GMP-P(NH)P enhances the Na+ effect. Moreover, the binding kinetic to this high-affinity state was also modified by Na+ and nucleotides. All of these data support the existence of two independent mu and delta binding sites, the properties of which are differentially regulated by these endogenous effectors. PMID- 2985759 TI - Characterization of a labile naloxone binding site (lambda site) in rat brain. AB - A high-affinity binding site selective for naloxone and other 4,5-epoxymorphinans (lambda site) has been previously described in rat brain. Following homogenization of freshly dissected brain, the lambda sites convert from a high affinity to a low-affinity state. When measured with [3H]naloxone, the decay is very rapid at 20 degrees C (t 1/2 less than 2 min), whereas it is progressively slowed at lower temperatures. Proteinase inhibitors, antoxidants, and sulfhydryl group-protecting agents failed to prevent this conversion. Kinetic measurements of mu and lambda binding at varying temperatures demonstrated that the decrease in lambda binding does not coincide with the concurrent increase in mu binding and that the loss of high-affinity lambda binding at 20 degrees C can be partially restored when the temperature is lowered to 0 degrees C. The low affinity state of the lambda site is rather stable in the Tris buffer homogenates and is susceptible to digestion by a protease. The (-)-isomer of WIN 44,441, a benzomorphan drug, binds to lambda sites with moderate affinity (dissociation constant, KD = 63 nM), whereas the (+)-isomer does not (KD greater than 10,000 nM), thus establishing stereoselectivity of the binding process. Neither the high affinity nor the low-affinity state of lambda binding is significantly affected by the presence of 100 mM sodium chloride or 50 microM Gpp(NH)p, (a GTP analog), which is in contrast to the dramatic effect of these agents on the established opioid receptor system. Naltrexone, naloxone, nalorphine, and morphine (in this order of decreasing potency) bind to the lambda site in vivo in intact rat brain over dosage ranges that are commonly employed in pharmacological studies. PMID- 2985758 TI - Solubilization and characterization of rat brain alpha 2-adrenergic receptor. AB - alpha 2-Adrenergic receptors labelled by [3H]-clonidine (alpha 2-agonist) can be solubilized from the rat brain in a form sensitive to guanine nucleotides with a zwitterionic detergent, 3-[3-(cholamidopropyl)-dimethylammonio]-1-propane sulfonate (CHAPS). About 40% of the original [3H]CLO binding sites in the membranes were solubilized with 6 mM CHAPS. Separation of the soluble [3H]CLO bound complex was performed by the vacuum filtration method using polyethylenimine-treated GF/B filters. Solubilized [3H]CLO binding sites retained the same pharmacological characteristics of membrane-bound alpha 2-adrenergic receptors. Scatchard plots of [3H]CLO binding to solubilized alpha 2-receptors were curvilinear, indicating the existence of the two distinct binding components. Solubilized receptors were eluted as a single peak from Bio-Gel A-1.5 m column with a Stokes radius of 6.6 nm. The isoelectric point was 5.6-5.8. Regulations of the receptor binding by guanine nucleotides, monovalent cations, and sulfhydryl-reactive agents were maintained intact in the soluble state, whereas those by divalent cations were lost. The apparent retention of receptors and guanine nucleotide binding regulatory component(s) in the soluble state may allow a investigation of the regulation mechanisms of the brain alpha 2 adrenergic receptor system at the molecular level. PMID- 2985760 TI - Separation of neuropeptides by HPLC: evaluation of different supports, with analytical and preparative applications to human and porcine neurophysins, beta lipotropin, adrenocorticotropic hormone, and beta-endorphin. AB - The separation of 38 hypophyseal or hypothalamic peptides and proteins by reversed-phase HPLC is described. The efficacy of several supports is discussed: large-pore reversed-phase silica supports turned out to be most effective in the separation of neuropeptides and proteins. The use of high sodium phosphate and phosphoric acid in the eluent resulted in high yields and better separation. On the basis of this procedure, beta-lipotropin, beta-endorphin, and adrenocorticotropic hormone were prepared from porcine pituitary glands, and human neurophysins from human pituitary posterior lobes. Moreover, this system is used for the analysis of the perifusate of pituitary posterior lobes, showing the release of micro-heterogeneous neurophysins. PMID- 2985761 TI - Characterization of "high-affinity" [3H]ouabain binding in the rat central nervous system. AB - The characteristics of [3H]ouabain binding were examined in various areas of rat brain. In the striatum, Scatchard analysis revealed a single class of "high affinity" binding sites with an apparent binding affinity (KD) of 10.4 +/- 0.9 nM and an estimated binding capacity (Bmax) of 7.6 +/- 1.9 pmol/mg protein. Similar monophasic Scatchard plots were found in the brainstem, cerebellum, hypothalamus, and frontal cerebral cortex. [3H]Ouabain binding to rat brain was sodium- and ATP dependent and strongly inhibited by potassium. Proscillariden A was the most potent cardiac glycoside tested in inhibiting specific [3H]ouabain binding to brain membranes, and the rank order of inhibitory potencies for a series of cardiac glycosides was similar to that previously reported for inhibition of heart Na,K-ATPase. To assess whether the high-affinity binding sites for [3H]ouabain were localized to neuronal or nonneuronal membranes, the effect of discrete kainic acid lesions on striatal [3H]ouabain binding was examined. Kainic acid lesions of the striatum reduced [3H]ouabain binding to striatal homogenates by 79.6 +/- 1.6%. This suggests that the "high-affinity" [3H]ouabain binding sites measured in our experiments are localized to neuronal elements. Thus, the high-affinity binding of [3H]ouabain to brain membranes may selectively label a neuronal form or conformation of Na,K-ATPase. PMID- 2985762 TI - Cholinergic-adrenergic receptor interactions in cerebral microvessels. AB - Enriched capillary preparations isolated from rat cerebral cortex were used to evaluate cholinergic-adrenergic receptor interactions in cerebral endothelium. Possible receptor interactions were determined by measuring an intracellular mediator, cyclic AMP and alterations in GTP-sensitive agonist binding. Unstimulated microvessel homogenates generate 66 +/- 16 pmol/mg/10 min of cyclic AMP. Adrenergic agonists norepinephrine and isoproterenol increase cyclic AMP to 147 +/- 31 and 149 +/- 23 pmol/mg/10 min, respectively. Addition of the muscarinic agonist carbachol has no effect on basal cyclic AMP but it completely blocks the stimulation elicited by adrenergic agonists. The displacement of quinuclidinyl benzilate (QNB) by carbachol yields an IC50 of 1.5 +/- 0.45 X 10( 4) M and a Hill coefficient of 0.54 +/- 0.07, indicating a heterogeneous population of binding sites. Guanine nucleotides shift the displacement curve to the right (IC50, 4.7 +/- 0.16 X 10(-4) M) and convert the binding site population to greater homogeneity (0.76 +/- 0.18). Isoproterenol prevents both the affinity shift and binding site conversion evoked by guanine nucleotides. These data suggest that cholinergic-adrenergic interactions occur at both the level of receptor binding and the generation of an intracellular messenger. Since cyclic AMP has been purported to play a role in regulation of blood-brain barrier permeability, the existence of adrenergic-cholinergic, i.e., excitatory inhibitory modulators of adenylate cyclase in cerebral endothelium, suggests that these receptors may mediate physiological and/or pathological alterations of cerebrovascular permeability. PMID- 2985763 TI - Distribution of glucose-1,6-bisphosphate and IMP-activated glucose bisphosphatase in brain and retina. AB - The activity of glucose-1,6-bisphosphatase and the level of its substrate were measured in 16 gray areas and four fiber areas of mouse brain and 10 layers or sublayers of monkey retina. Because of the low activity of the enzyme and the small sample sizes, it was necessary to develop a method with two different amplification steps (overall amplification about 10(6]. The enzyme ranged in activity 100-fold from a low in monkey retina photoreceptor cells to a high in the pyramidal layer of the hippocampus. However, in gray areas of the brain proper the range was only about fourfold. This, together with its requirement for IMP, suggests that the enzyme has a widespread metabolic function related to states of increased neuronal activity. Glucose-1,6-bisphosphate levels varied from 80 to 960 mu mol/kg dry weight in different areas of mouse brain and from 44 to 200 mu mol/kg dry weight in different layers of monkey retina. In general, the glucose bisphosphate levels correlated positively with the bisphosphatase activities; however, the three areas with the highest enzyme concentrations did not fit this pattern. PMID- 2985764 TI - Taurine stimulation of isolated hamster brain Na+,K+-ATPase: activation kinetics and chemical specificity. AB - Epileptic foci are associated with locally reduced taurine (2-aminoethanesulfonic acid) concentration and Na+,K+-ATPase (EC 3.6.1.3) specific activity. Topically applied and intraperitoneally administered taurine can prevent the development and/or spread of foci in many animal models. Taurine has been implicated as a possible cytosolic modulator of monovalent ion distribution, cytosolic "free" calcium activity, and neuronal excitability. Taurine may act in part by modulating Na+,K+-ATPase activity of neuronal and glial cells. We characterized the requirements for in vitro modulation of Na+,K+-ATPase by taurine. Normal whole brain homogenate Na+,K+-ATPase activity is 5.1 +/- 0.4 (4) mumol Pi X h-1 X mg-1 Lowry protein. Partial purification of the plasma membrane fraction to remove cytosolic proteins and extrinsic proteins and to uncouple cholinergic receptors yields a membrane-bound Na+,K+-ATPase activity of 204.6 +/- 5.8 (4) mol Pi X h-1 X mg-1 Lowry protein. Taurine activates the Na+,K+-ATPase at all levels of purification. The concentration dependence of activation follows normal saturation kinetics (K1/2 = 39 mM taurine, activation maximum = +87%). The activation exhibits chemical specificity among the taurine analogues and metabolites: taurine = isethionic acid greater than hypotaurine greater than no activation = beta-alanine = methionine = choline = leucine. Taurine can act as an endogenous activator/modulator of Na+,K+-ATPase. Its action is mediated by a membrane-bound protein. PMID- 2985766 TI - Regional mitochondrial respiratory activity in Huntington's disease brain. AB - This study investigated mitochondrial respiratory activity in Huntington's disease (HD) brain. Mitochondrial membranes from caudate and cortex of HD and non HD autopsied brains were assayed for succinate oxidation, cytochrome oxidase activity, and cytochromes b, cc1, and aa3. There was a significant decrease in HD caudate mitochondrial respiration, cytochrome oxidase activity, and cytochrome aa3, whereas cytochromes b and cc1 were normal. These findings are consistent with the hypothesis that mitochondrial dysfunction may contribute to the localized hypometabolism and progressive atrophy of the HD caudate. PMID- 2985765 TI - Binding sites for L-[3H]glutamate on hippocampal synaptic membranes: three populations differentially affected by chloride and calcium ions. AB - The effects of Cl- and Ca2+ were studied on the specific binding of L [3H]glutamate to multiple sites on rat hippocampal synaptic membranes. Quisqualate (5 microM) or DL-2-amino-4-phosphonobutyrate (2-APB) (300 microM) was used to discriminate two previously identified classes of binding sites. Saturation isotherms and displacement curves constructed under different ionic conditions suggested that the effects of Cl- and Ca2+ could best be explained by postulating the existence of three major binding site populations in this preparation rather than two. The binding of L-glutamate to Glu A sites exhibits an absolute dependence on Cl-, and Ca2+ markedly increases the maximum density of these sites. Glu A sites bind quisqualate and 2-APB with relatively high affinity. Cl- (47 mM) more than doubles the maximum density of Glu B sites, but Ca2+ appears to have no effect. Glu B sites can be discriminated from the other classes by their relatively low affinity for quisqualate and 2-APB. There is reason to think that the Glu B population is heterogeneous. The novel Glu C population can be virtually selectively labeled by exposing 2-APB-sensitive binding sites to radioligand in Tris-HOAc buffer with Ca2+. Binding of L [3H]glutamate to these sites is enhanced by both Cl- and Ca2+, but requires neither ion. Ca2+ appears to increase both the affinity of Glu C sites for L glutamate and their maximum binding site density. In the presence of Ca2+ and Cl , Glu C sites bind the radioligand with micromolar affinity (KD approximately 2 microM) and high capacity (Bmax approximately 160 pmol/mg protein).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2985767 TI - Some effects of prolonged constriction on nerve regeneration in the rabbit. AB - Serial nerve conduction studies have been used to study the time-course of regeneration after tibial nerve crush in the rabbit, and the effect on this of a constricting ligature 5 mm distal to the site of crush. Transverse sections from the region of the ligature showed that when constriction was severe, the regenerating fibers had difficulty in penetrating the constricted region, and that some took an aberrant course in superficial connective tissue outside the ligature. In 2 out of 6 animals no fibres succeeded in passing through the centre of ligature, but aberrant fibres were able to re-enter distal fascicles and to reinnervate distal muscles. When compared with findings after crush alone, the time taken to reinnervate distal muscles was increased by a ligature, and distal fibre diameter and motor conduction velocity remained low. Low voltage, long duration, polyphasic muscle action potentials were evoked by distal nerve stimulation, and axon reflexes suggested excessive axon branching within nerve trunks. Since these changes were seen in the distal parts of both constricted and aberrant fibres, they cannot be attributed to constriction alone. Similar changes might be expected in other peripheral nerve lesions in which loss of fascicular continuity and neuroma formation occur. PMID- 2985768 TI - Thermographic presentation of cutaneous sensory and vasomotor activity in the injured peripheral nerve. AB - Impaired function of cutaneous segments of monkey peripheral nerves experimentally blocked by lidocaine anesthesia was clearly visualized by means of elevated temperature measurements obtained on computerized color telethermography. Mean temperature elevations in the segments of anesthetized primate nerves were 2.40 degrees C at the ulnar segment 17 minutes after nerve block, and 1.20 degrees C at the peroneal nerve at 20 minutes. The vasomotor activity of specific nerves, recorded after local anesthesia and displayed by color telethermographic imaging, corresponded to the distribution of sensory segments identified by more cumbersome means. Telethermography is therefore shown to be a useful tool, both qualitatively and quantitatively, in mapping cutaneous distribution of peripheral nerves and for evaluation of peripheral nerve injuries. PMID- 2985769 TI - Thermographic imaging of cutaneous sensory segment in patients with peripheral nerve injury. Skin-temperature stability between sides of the body. AB - Sensory examination based on the patient's subjective assessment of symptoms may raise difficult questions about whether the individual's expressed complaint is based on organic nerve damage, psychogenic factors, or even malingering. A prototype computerized telethermograph has allowed clinical quantification of peripheral nerve injury. The system makes possible mapping and imaging of the damaged area, as well as skin temperature measurements. In normal persons, the skin temperature difference between sides of the body was only 0.24 degree +/- 0.073 degree C. In contrast, in patients with peripheral nerve injury, the temperature of the skin innervated by the damaged nerve deviated an average of 1.55 degrees C (p less than 0.001). The new technique requires further refinement, but it appears that use of this method may be cost-effective in helping to resolve medicolegal conflicts concerning peripheral nerve injury. PMID- 2985770 TI - Role of monoamines in experimental spinal cord injury in rats. Relationship between Na+-K+-ATPase and lipid peroxidation. AB - A spinal cord injury was produced in Wistar rats by extradural compression of the cord with a Sugita aneurysm clip for 5 seconds. During a 2-week observation period following the injury, the tissue norepinephrine (NE), dopamine (DA), and serotonin (5-HT) concentrations decreased uniformly at and below the injured site. The chemical denervation of NE or 5-HT neurons produced by the intraspinal injection of 6-hydroxydopamine (6-OHDA) or 5,7-dihydroxytryptamine (5,7-DHT) 2 weeks before the injury did not cause a marked difference in the extent of hemorrhagic necrosis of the spinal cord after trauma as compared to control animals without pretreatment. In the rats pretreated with 6-OHDA, NE was decreased to less than 30% of control (non-pretreated) values, and, beginning at 5 days after injury, motor performance (assessed quantitatively with the inclined plane method) was significantly improved compared to results in the non pretreated control rats. The rats pretreated with 5,7-DHT showed no change from control animals. Spinal cord samples from non-pretreated control animals obtained at the injury site 30 minutes after the compression injury showed a marked decrease in the activity of synaptosomal Na+-K+-ATPase (adenosine triphosphatase) of about 50%, and an increase in both thiobarbituric acid reaction substance (about 170%) and cyclic guanine monophosphate (about 150%). The NE-denervated rats showed no significant changes in these three parameters. The results indicated that NE released after crush injury may impair the neuronal cell membrane around the lesion site by induction of lipid peroxidation. The possible mechanisms by which released NE may alter membrane function are discussed. PMID- 2985771 TI - Diagnosis and treatment of jaw tumors in children. AB - A retrospective review of experience with jaw tumors in 48 children over a ten year period was made. These were predominantly benign, nonodontogenic, mesenchymal tumors. Clinical behavior was variable and often did not correspond to the histologic patterns. The clinically aggressive variants displayed rapid growth usually with associated pain. Resection rather than curettage is appropriate therapy when there has been earlier treatment failure by local tumor removal or rapid tumor expansion, or when tumor extensively invades the jaw. When resection is carried out, facial growth is not adversely affected if early reconstruction with autogenous bone is performed. PMID- 2985772 TI - Intolerance to aspirin and the nonsteroidal anti-inflammatory drugs. AB - A constant enigma has been the ability of aspirin and other structurally unrelated nonsteroidal anti-inflammatory drugs to induce non-IgE mediated allergic reactions. These reactions range from mild hypersensitivity to fatal anaphylaxis. Recent biochemical and pharmacologic studies involving the oxidative metabolism of arachidonic acid in different cells and tissues have provided insights into how this could conceivably occur. The products of cyclo-oxygenase and lipoxygenase pathways of arachidonic acid metabolism and their interactions may provide an approach, if not the solution, to the problem of aspirin intolerance. PMID- 2985773 TI - Vitamin D and skeletal tissues. AB - It is now accepted that vitamin D is an integral part of a complex endocrine system, one with far-reaching implications in mineral metabolism. Reviews of the sources, functions and metabolism of vitamin D, as currently understood, are presented as a prelude to discussions of the role of vitamin D in calcium and phosphorous homeostatis and possible specific roles for vitamin D in mineralized tissues. Data describing a possible regulatory function for vitamin D in bone and bone protein metabolism are presented. Some of the controversy which presently exists regarding the biochemical mechanism of the action of this vitamin is discussed. Finally, the possible relationship of vitamin D and disorders of skeletal tissues is described. PMID- 2985774 TI - Disposition and bioavailability of various formulations of tetrahydrocannabinol in the rhesus monkey. AB - Oral delta 9-tetrahydrocannabinol (THC) in gelatin capsules is under evaluation as an antiemetic agent in cancer patients, but knowledge concerning its bioavailability is incomplete and, furthermore, alternative routes of administration may be desirable. In this study, the disposition of THC was determined in four rhesus monkeys given 2.5-mg/kg doses using the following routes of administration and formulations: intravenous (iv); orally (po) on a cookie and in gelatin capsules; intramuscularly (im) in Tween-80 and in Emulphor EL620; rectally in various suppository bases. Serum THC concentrations were measured by RIA and analyzed by weighted nonlinear regression. Serum concentrations were best described by a sum of two exponentials with alpha and beta half-lives (mean +/- SD) of 0.74 +/- 0.59 and 14.9 +/- 12.5 h. Apparent bioavailability (%F +/- SD) of various formulations of THC were: gelatin capsules, 26 +/- 14; cookie, 89 +/- 16; intramuscularly in Tween-80 and in Emulphor, 39 +/- 13 and 102 +/- 15, respectively. Using the method of statistical moments, mean residence times in the body (h +/- SD) were: intravenous, 6.08 +/- 1.60; cookie, 21.92 +/- 3.11; gelatin capsule, 26.80 +/- 23.61; intramuscularly in Emulphor, 10.92 +/- 3.46 (in Tween-80, not calculated). THC was not bioavailable by the rectal route. We conclude from this study that THC formulated as a gelatin capsule exhibits a low and variable extent of bioavailability and that intramuscular THC may be a useful alternative route of administration since it is more completely bioavailable. PMID- 2985776 TI - Prevalence and persistence of different strains of Candida albicans in treatment of denture stomatitis. PMID- 2985775 TI - Calcium-dependent chloride secretion in rat colon epithelium. AB - Epithelia, dissected from the descending rat colon, were studied under short circuit conditions in Ussing chambers. The latter were modified to accept flexible light guides, so that the tissue could be irradiated, with white light, normal to its surface. Irradiation alone had no effect on short-circuit current (s.c.c.). In the presence of erythrosine B (which by itself had no effect on the s.c.c.) on the basolateral side of the tissue, irradiation produced a substantial increase in s.c.c.; this increase was sustained after irradiation had ceased and the dye had been washed away. The photodynamic effect of erythrosine B required the presence of oxygen in the bathing solution. Also calcium was essential for the s.c.c. response to occur. Thus, irradiation in the presence of the dye in the absence of calcium had no effect on s.c.c., but a s.c.c. increase could be revealed by subsequent addition of calcium after irradiation had ceased. Cobalt and magnesium ions antagonized the effect of calcium in the conditions described above. Ion flux measurements with 36Cl and 22Na showed that the photodynamic effect of erythrosine B abolished net sodium absorption and reversed net chloride absorption to secretion. The data are consistent with abolition of electroneutral sodium chloride absorption and the stimulation of electrogenic chloride secretion to an extent equivalent to the s.c.c. responses. Using calcium-containing buffers it was possible to compare the s.c.c. responses at low, known ionized calcium concentrations with the maximal chloride secretory effect following photodynamic activation. Chloride secretion was half-maximally activated when the basolateral bathing fluid contained 1 microM-ionized calcium and after the basolateral face of the tissue had been permeabilized by the photodynamic action of erythrosine B. The relation between ionized calcium concentration in the basolateral fluid and the chloride secretory response was steep. PMID- 2985777 TI - Enhanced signal transduction by adenylate cyclase in platelet membranes of patients showing antidepressant responses to alprazolam: preliminary data. AB - The triazolobenzodiazepine, alprazolam, was administered to 11 depressed patients over a period of six weeks, and six patients showed a favorable antidepressant response. There were no significant differences between responders and nonresponders in age, pretreatment Hamilton Depression Rating Scores, 4 p.m. postdexamethasone plasma cortisol levels, or platelet monoamine oxidase activities. Blood levels of alprazolam were not meaningfully different in responders and nonresponders when measured on treatment day 8. However, on treatment day 8, significantly enhanced prostaglandin D2-stimulated platelet adenylate cyclase activity, greater suppression of prostaglandin D2-stimulated platelet adenylate cyclase activity by epinephrine, and enhanced sodium fluoride stimulated platelet adenylate cyclase activity were seen in the six patients who went on to respond to alprazolam, but not in the five nonresponders. In contrast, there were no significant changes in prostaglandin D2, (-)-isoproterenol, or fluoride ion-stimulated leukocyte adenylate cyclase activity in responders or nonresponders. No meaningful changes were observed in the mean densities of either the high-affinity platelet alpha 2-adrenergic receptor (for 3H-p aminoclonidine) or the leukocyte beta-adrenergic receptor (for 3H dihydroalprenolol) in responders or nonresponders. The present findings, taken in conjunction with findings from other recent studies, suggest that enhanced coupling between certain neurotransmitter or hormone receptors and adenylate cyclase through the guanine nucleotide regulatory proteins may help explain the antidepressant effects of alprazolam and possibly other forms of antidepressant treatment. PMID- 2985778 TI - Direct evidence for de-novo synthesis of LH receptors in cultured pig granulosa cells in response to FSH. AB - FSH plus insulin, cortisol, and thyroxine (IFT) stimulated incorporation of dense isotope-containing (2H, 13C, 15N) amino acids into soluble 125I-labelled hCG binding sites. Evidence of new synthesis of binding sites appeared as early as 3 h after the beginning of the pulse-labelling period. By 48 h the majority of detectable soluble 125I-labelled hCG binding sites appeared to be newly synthesized. Studies with FSH + IFT and puromycin indicated that FSH + IFT stimulated synthesis of new LH/hCG binding sites, and that internalization or degradation of LH/hCG binding sites may also require protein synthesis. PMID- 2985779 TI - Effects of different numbers of ectopic pituitary transplants on regulation of testicular LH/hCG and prolactin receptors in the hamster (Mesocricetus auratus). AB - Adult male hamsters were given transplants of 1/2, 1, 2, 3 or 4 pituitaries under the kidney capsule and were killed 4 weeks later. Pituitary transplants produced a significant, dose-related increase in plasma prolactin levels, no changes in plasma LH and an increase in plasma FSH. Concentration of LH/hCG receptors in the testes was significantly increased in animals with 2 or 3 transplants and concentration of testicular prolactin receptors was significantly increased in those given 2 transplants. The apparent stimulatory effects of 1/2, 1 or 4 transplants on testicular LH/hCG and prolactin binding were not statistically significant. Some of the animals were injected with 0.3 i.u. hCG/g body weight 24 h before being killed. This produced a significant reduction in the levels of prolactin receptors and an apparent reduction in the levels of LH/hCG receptors in the testes. Elevation of plasma testosterone concentrations in response to hCG was significantly greater in animals given 3 or 4 pituitary transplants than in the remaining groups. These results provide further evidence that prolactin increases the number of LH/hCG and prolactin receptors in the hamster testis and suggest that changing the number of ectopic pituitary transplants may result in biphasic effects on the testis, with 2 or 3 transplants being maximally stimulatory. PMID- 2985780 TI - Changes in pituitary and ovarian LHRH receptors after active immunization of female rats against LH or LHRH. AB - The hypothalamic-pituitary-ovarian axis of female rats was disrupted at the site of LHRH stimulation by active immunization against LHRH or at the site of LH action by active immunization against LH. Active immunization against LH was associated with an increase in pituitary LHRH receptors to levels comparable to control values at pro-oestrus whereas immunization against LHRH led to a marked reduction in receptor numbers. Ovarian LHRH receptor concentrations were increased by both treatments. It is concluded, therefore, that (1) LHRH receptors in the pituitary and ovary are not concomitantly controlled, and (2) pituitary receptor numbers are primarily under positive autoregulatory control by LHRH and that ovarian LHRH receptor concentrations may be under long-term influence of LH. PMID- 2985781 TI - A new generation of phosphodiesterase inhibitors: multiple molecular forms of phosphodiesterase and the potential for drug selectivity. AB - With several notable exceptions, interest in the area of multiple molecular forms of phosphodiesterase remained relatively dormant during the decade following Thompson's discovery of more than one phosphodiesterase in brain in 1971. Within the last several years, however, over 20 novel agents have been identified that exert selective inhibitory effects on the various molecular forms of phosphodiesterase present within different cells. In addition, several studies have documented that such agents can produce discrete changes in cyclic AMP and cyclic GMP, an action that is not shared by "first generation" phosphodiesterase inhibitors such as theophylline. The purpose of this Perspective is to provide some clarity to this rapidly evolving area of selective phosphodiesterase inhibitors. Thus, we have attempted to characterize the different forms of phosphodiesterase present in various tissues and cells according to their kinetic properties, substrate specificity, etc. and also to characterize those major classes of agents that have been shown to inhibit phosphodiesterase activity, whether selectively or nonselectively. In addition, we have described several therapeutic areas wherein selective phosphodiesterase inhibitors might prove efficacious, paying particular attention to those areas in which selective phosphodiesterase inhibitors have already been shown to exert beneficial effects, namely, stimulation of myocardial contractility, inhibition of mediator release, and inhibition of platelet aggregation. Although focusing on these three areas, it is obvious that the potential therapeutic utility of selective phosphodiesterase inhibitors could conceivably extend to several other areas in which modulation of cyclic nucleotides can have desirable effects, including cancer chemotherapy, analgesia, the treatment of depression, Parkinson's disease, and learning and memory disorders. For example, the selective type III phosphodiesterase inhibitor rolipram has been shown to antagonize reserpine induced hypothermia and also to potentiate yohimbine lethality, two tests that are indicative of antidepressant activity. In addition, microinjection of the selective PDE III inhibitor Ro 20-1724 into the rat brain stem has been shown to produce analgesia.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985782 TI - Synthesis and antiviral activity of the carbocyclic analogues of (E)-5-(2 halovinyl)-2'-deoxyuridines and (E)-5-(2-halovinyl)-2'-deoxycytidines. AB - The carbocyclic analogues of the potent and selective antiherpes agents (E)-5-(2 bromovinyl)-2'-deoxyuridine (BVDU), (E)-5-(2-iodovinyl)-2'-deoxyuridine (IVDU), and (E)-5-(2-bromovinyl)-2'-deoxycytidine (BVDC) were synthesized by conventional methods with use of carbocyclic 2'-deoxyuridine as starting material. C-BVDU, C IVDU, and C-BVDC were equally selective, albeit slightly less potent, in their antiherpes action than BVDU, IVDU, and BVDC. Although resistant to degradation by pyrimidine nucleoside phosphorylases, C-BVDU did not prove more effective than BVDU in the systemic (oral, intraperitoneal) or topical treatment of HSV-1 infections in mice. PMID- 2985783 TI - Cyclic melanotropins. 9. 7-D-Phenylalanine analogues of the active-site sequence. AB - The cyclic melanotropin Ac-Ser1-Tyr2-Ser3-Cys4-Glu5-His6-Phe7-Arg8 -Trp9-Cys10 Lys11-Pro12-Val13-NH is a highly potent agonist as determined in several melanocyte bioassays. In linear melanotropins, a D-Phe7 substitution leads to increased potency and often prolonged biological activity. In order to determine if this substitution would have the same effect in cyclic melanotropins, we have prepared a series of these analogues. The D-Phe7-substituted cyclic melanotropins Ac-[Cys4,D-Phe7,Cys10]-alpha-MSH4-10-NH2 and Ac-[Cys4,D-Phe7,Cys10]-alpha-MSH4-11 NH2 were both more potent than their cyclic L-Phe7-containing counterparts in either the frog or lizard skin bioassay by more than a factor of 10. Neither peptide, however, exhibited prolongation of biological activity in either assay. Substitution of D-Phe7 into the cyclic 4-12 and 4-13 sequences led to a slight or no increase in potency in both assays relative to the L-Phe7 counterparts, but the activity of the melanotropins was ultraprolonged in each assay. Ac-[Cys4,D Phe7,Cys10]-alpha-MSH4-12-NH2 was about equipotent to Ac-[Cys4,D-Phe7,Cys10] alpha-MSH4-13-NH2, again demonstrating, as with certain linear and cyclic L-Phe7 containing melanotropins, that the C-terminal amino acid valine is not required for biological activity or for superpotency. Similar to the linear D-Phe7 analogues that possessed ultraprolonged melanotropic activity, the 4-12 and 4-13 cyclic D-Phe7 analogues also displayed the phenomenon of superagonism, which is a time-dependent increase in efficacy over that produced by an equipotent concentration of the native hormone. Cyclization of certain linear melanotropins resulted in analogues with increased resistance to biological degradation by serum enzymes or purified proteolytic enzymes. Further, incorporation of a D-Phe7 into in the cyclic analogues led to melanotropins that were totally resistant to enzymatic inactivation by trypsin. PMID- 2985784 TI - "Reverse" and "symmetrical" analogues of actinomycin D: metabolic activation and in vitro and in vivo tumor growth inhibitory activities. AB - Two new classes of actinomycin D analogues, tetracyclic "reverse" analogues and a tricyclic "symmetrical" analogue of actinomycin D, are reported. These analogues bind to DNA and the binding does not occur by an intercalation mechanism. The analogues inhibit the synthesis of DNA and RNA in P388 tumor cells and the growth of CCRF-CEM cells in vitro at nanomolar concentrations. The tetracyclic "reverse" analogues, which are structurally related to the previously reported actinomycin D oxazolyl analogues, are metabolized in the presence of rat hepatic microsomes and tumor cell homogenates. The metabolism takes place with the loss of the oxazole ring; thus the "reverse" analogues produce a major metabolite which is the "symmetrical" analogue; the actinomycin oxazolyl analogues generate 7 hydroxyactinomycin D. Further, the microsomes activate the analogues to free radical states which catalyze the production of superoxide as shown by stimulation of epinephrine oxidation and also indicated by electron paramagnetic resonance studies. The "symmetrical" and "reverse" analogues also demonstrate very high activities in these systems. In in vivo studies using P388/S, P388/ADR leukemia, and B16 melanoma in mice, the analogues showed increased activity and superior therapeutic index values, in comparison to actinomycin D. PMID- 2985785 TI - Orally active and potent inhibitors of gamma-aminobutyric acid uptake. AB - 3-Pyrrolidineacetic acid (1a), certain piperidinecarboxylic acids--i.e., 3 piperidinecarboxylic acid (2a), 1,2,5,6-tetrahydro-3-pyridinecarboxylic acid (3a), and cis-4-hydroxy-3-piperidinecarboxylic acid (4a)--cis-3 aminocyclohexanecarboxylic acid (5a, cis-3-ACHC), and gamma-aminobutyric acid (6a, GABA) itself are among the most potent inhibitors of [3H]GABA uptake by neurons and glia in vitro. These hydrophilic amino acids, however, do not readily enter the central nervous system in pharmacologically significant amounts following peripheral administration. We now report that N-(4,4-diphenyl-3 butenyl)-3-piperidinecarboxylic acid (2b) is a specific GABA-uptake inhibitor that is more potent, more lipophilic and, in limited testing, as selective as 2a. Similar results were obtained with the N-(4,4-diphenyl-3-butenyl) derivatives of 1a, 3a, and 4a. By contrast, N-(4,4-diphenyl-3-butenyl) derivatives of 5a and 6a were not more potent than the parent amino acids and appear to inhibit GABA uptake, at least in part, by a nonselective mechanism of action. The N-(4,4 diphenyl-3-butenyl)amino acids 1b-4b exhibit anticonvulsant activity in rodents following oral or intraperitoneal administration [Yunger, L.M.; et al. J. Pharmacol. Exp. Ther. 1984, 228, 109]. PMID- 2985786 TI - Synthesis and structure-activity studies on excitatory amino acids structurally related to ibotenic acid. AB - With use of ibotenic acid as a lead, analogues of (RS)-alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA) and of (RS)-3-hydroxy-4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridine-7-carboxylic acid (7-HPCA) were synthesized and tested as excitants of neurons in the cat spinal cord by using microelectrophoretic techniques and as inhibitors of the binding of kainic acid in vitro. Like AMPA and 7-HPCA, (RS)-3-hydroxy-4,5,6,7-tetrahydroisoxazolo[5,4-c] pyridine-5-carboxylic acid (10, 5-HPCA) and (RS)-3-hydroxy-5 (bromomethyl)isoxazole-4-propionic acid (11, ABPA) proved to interact potently and selectively with central quisqualic acid receptors, assumed to represent physiological glutamic acid receptors. Analogues of 7-HPCA or 10, in which one or both of the acid groups were masked, were very weak or inactive as neuronal excitants and had no antagonistic effects at excitatory amino acid receptors. The structure of 7-HPCA in the crystalline state was established by X-ray analyses. The preferred conformation of 10 in aqueous solution was determined by 1H NMR spectroscopy. On the basis of these studies, 7-HPCA as well as 10 were shown to adopt preferentially conformations with the carboxylate groups in equatorial positions. It is suggested that AMPA, 7-HPCA, and 10 interact with quisqualic acid receptors in conformations essentially reflecting active conformation(s) of glutamic acid at these receptors. PMID- 2985788 TI - T cell early activation antigens expressed by peripheral lymphocytes in Crohn's disease. AB - We studied the expression of early activation antigens (4F2, transferrin receptor, IL-2 receptor) on peripheral lymphocytes (PBL) of patients with Crohn's Disease. We have found that the proportion of PBL expressing these antigens was significantly higher in patients than in controls. The expression of the 4F2 antigen was more pronounced than that of other activation antigens directly involved in promoting cell growth (e.g. transferrin receptor, IL-2 receptor). These results indicate that CD patients have an increased number of T cells in a very early phase of activation. PMID- 2985787 TI - Synthesis and interaction of 5-(substituted-phenyl)-3-methyl-6,7 dihydropyrazolo[4,3-e] [1,4]diazepin-8(7H)-ones with benzodiazepine receptors in rat cerebral cortex. AB - On the basis of the anxiolytic property of ripazepam, 1-ethyl-4,6-dihydro-3- methyl-8-phenylpyrazolo[4,3-e][1,4]diazepin-5(1H)-one (1), a series of isomeric 5 (phenyl-substituted)pyrazolo[4,3-e][1,4] diazepin-8-ones 3a-f were prepared and tested for their ability to bind to the benzodiazepine receptor. All compounds 3a f display affinities for the benzodiazepine receptor in the microM range of concentration; in particular 5-phenyl-3-methyl-6,7-dihydropyrazolo[4,3-e][1,4] diazepin-8(7H)-one (3a) is 2 orders of magnitude less potent in inhibiting [3H]flunitrazepam binding than diazepam and displays an affinity for the benzodiazepine receptor practically comparable to that of its structural isomer, ripazepam, and to that of chloriazepoxide. PMID- 2985789 TI - Virus myocarditis: molecular hybridization allows the detection of virus-RNA in heart muscle after virus infection. AB - In most patients with virus myocarditis, the diagnosis is still based on clinical data alone. Endomyocardial biopsies subjected to electron microscopy, immunofluorescence techniques and virus isolation procedures provide additional, but only occasionally conclusive information. In this communication we describe a new method which could possibly be used to improve the diagnostic possibilities in patients with suspected virus myocarditis. The method is based on the hybridization of radioactive complementary nucleotide sequences to virus-RNA. It is shown that in an experimental model (reovirus infected baby mice) this method can be used to demonstrate the virus infection of cardiac muscle. It is suggested that the method could be adapted to other viruses (e.g. coxsackie virus) and to endomyocardial biopsies derived from patients with suspected virus myocarditis. PMID- 2985790 TI - Calcium-activated neutral protease in the peripheral nerve, which requires microM order Ca2+, and its effect on the neurofilament triplet. AB - There are two types of calcium-activated neutral protease (CANP), m-CANP and mu CANP, following the nomenclature of Suzuki et al to show that each requires mM and microM Ca2+, respectively, for its activation. We found mu-CANP activity in a crude CANP fraction extracted from the peripheral nerve, which degraded the neurofilament (Nf) triplet (200 K, 160 K, 68 K), especially the 160 K component, at Ca2+ concentrations of 50 microM and 0.1 mM. The triplet was degraded in the order of the 160 K, 68 K, and 200 K components, respectively. In addition, the effects of partially purified mu-CANP of rabbit skeletal muscle, purified natural mu-CANP of bovine liver, derived mu-CANP prepared by autodigestion of chicken muscle m-CANP, m-CANP of chicken skeletal muscle, and cathepsin B of rat liver on the Nf were examined. Among the triplet components, the 160 K component was most rapidly degraded by all proteases so far tested. The difference in the effect of mu-CANP and m-CANP or cathepsin B on susceptibility of the 200 K component to degradation might be due to the difference of the relative amounts of enzymes to Nf. PMID- 2985791 TI - Cholinergic neurotransmission in the central nervous system of the Snell dwarf mouse. AB - An unequal decrease in cholinergic activity has been evidenced in discrete brain areas in the growth hormone, thyroid-stimulating hormone and prolactin deficient Snell dwarf mouse. The effect of the mutation's pituitary deficit on central cholinergic mechanisms appears to be selective: Normally high cholinergic activity areas such as striatum, olfactory tubercles, and hippocampus show strong alterations in this neurotransmitter system. Structures which appear earlier in ontogenesis are less affected, if at all. The lack of pituitary hormones seems to have effects on choline acetyltransferase activity and/or synthesis as well as on the development of high affinity (H.A.) cholinergic uptake mechanisms, both strongly defective in hippocampus and striatum. Therefore, a lower density of cholinergic terminals can be inferred. Furthermore, our observations are consistent with a close functional coupling of the choline H.A. transport and of subsequent choline acetylation. Acetylcholinesterase activity does not seem to be affected. Moreover, a compensatory effect at the postsynaptic level may have occurred due to developmental or functional plasticity for cholinergic responsiveness. In conclusion, the dwarf mouse seems to be a useful model for a better understanding of the influences of growth hormone and thyroid hormones on the development of central cholinergic mechanisms. It also provides the possibility to attempt a functional restoration of the deficient cholinergic neurotransmission and the behavioral disturbances which may be linked to them, by hormone replacement. PMID- 2985792 TI - The biology, pathophysiology and control of eicosanoids in inflammation. AB - The involvement in inflammatory conditions of those cyclo-oxygenase and lipoxygenase derivatives of arachidonic acid (5,8,11,14-eicosatetraenoic acid), which are known as the eicosanoids, is reviewed in the light of recent studies. Although it is now generally recognized that cyclo-oxygenase products are fundamental to the inflammatory process as chemical mediators, and that inhibition of the cyclo-oxygenase enzyme pathway explains the mode of action of most non-steroidal anti-inflammatory drugs (NSAIDs) commonly prescribed in veterinary practice, evidence for the involvement of lipoxygenase products of arachidonate metabolism in inflammation is increasing. The leukotrienes (LTs) are 5-lipoxygenase-derived eicosanoids which have been shown to be leucotactic and involved in anaphylactic and hypersensitivity reactions. Leucocytes, drawn to sites of injury by chemotaxis, themselves liberate pro-inflammatory eicosanoids which perpetuate the response and may aggravate the clinical condition. At therapeutic dose rates, most NSAIDs have no effect on the biosynthesis of LTs, whereas corticosteroids, by inhibiting the release of arachidonic acid, may prevent the formation of both cyclo-oxygenase and lipoxygenase products. However, because of the undesirable side-effects of steroids, the clinical use of these agents in treating inflammatory conditions is sometimes limited. Novel non steroid inhibitors of cyclo-oxygenase and lipoxygenase enzyme pathways could offer more effective and safer control of inflammation in animals. PMID- 2985793 TI - Molecular cloning and partial sequencing of hepatitis A viral cDNA. AB - Hepatitis A virus was purified from infected monkey kidney cell cultures, and the viral RNA was used to synthesize double-stranded cDNA. This cDNA was cloned either after insertion into a plasmid-primed synthesis system or after insertion into the PstI site of pBR322. The resulting clones were mapped by restriction endonuclease analysis and by cross hybridization of the viral inserts to generate a composite map which represented at least 97% of the viral genome, lacking ca. 220 bases from the 5' end of the genome. The clones were verified to be hepatitis A virus specific based on their positive hybridization to viral RNA and to total hepatitis A virus-infected cellular RNA from a heterologous marmoset host system. The nucleotide sequence of 3,054 base pairs of cDNA homologous to the 5' half of the viral genome was determined, and an open reading frame of 854 consecutive coding triplets was identified. In addition, sequences which encode the VP-1 and VP-3 viral structural proteins were located in the nucleotide sequence. PMID- 2985794 TI - Poliovirus RNA-dependent RNA polymerase and host cell protein synthesize product RNA twice the size of poliovirion RNA in vitro. AB - The poliovirus RNA-dependent RNA polymerase required an oligouridylate primer or a HeLa cell protein (host factor) to initiate RNA synthesis on poliovirion RNA in vitro. The polymerase synthesized template-sized product RNA in the oligouridylate-primed reaction. In the host factor-dependent reaction, the largest product RNA synthesized by the polymerase was twice the size of the template RNA. About half of the product RNA recovered from this reaction was shown to exist in the form of a snapback sequence. Time-course reactions and pulse-chase experiments showed that the product RNA was only slightly larger than the template RNA at early reaction times and that with time it increased in size to form the dimer-sized product RNA. Inhibition of the elongation reaction by adding only [alpha-32P]UTP and ATP resulted in the formation of template-sized product RNA. The dimer-sized product RNA was unaffected by phenol extraction or proteinase K treatment but was converted to template-sized molecules by S1 nuclease. Dimer-sized poliovirus RNA that was sensitive to S1 nuclease was also isolated from poliovirus-infected cells. The results from this study indicate that the labeled negative-strand product RNA synthesized in vitro was covalently linked to the positive-strand template RNA. Thus, in vitro, the primer-dependent poliovirus RNA polymerase may initiate RNA synthesis in the presence of the host factor by using the 3' end of the template RNA as a primer. PMID- 2985795 TI - Isolation by high-performance liquid chromatography and partial characterization of a 57,000-dalton herpes simplex virus type 1 polypeptide. AB - A Nonidet P-40 extract of HSV-1-purified virions was fractionated by reversed phase high-performance liquid chromatography (RP-HPLC). The first peak fraction eluted at 25% organic solvent. Polyacrylamide gel electrophoresis showed that it contained a 57,000-dalton polypeptide. The polypeptide was characterized by determination of the amino acid composition and the N-terminal amino acid sequence. Adsorption of the detergent extract before RP-HPLC showed that the polypeptide reacted with monoclonal antibodies LP1 directed against herpes simplex virus polypeptide VP-16. PMID- 2985796 TI - Reverse transcription of 7S L RNA by an avian retrovirus. AB - Linial et al. isolated a quail cell line, SE21Q1b, that is transformed by a single integrated provirus of Rous sarcoma virus. Virus particles are released from these cells, but because of a provirus defect, cellular rather than viral RNA is packaged. When these virus particles are disrupted with melittin in the presence of an appropriate reaction mixture containing actinomycin D, there is significant reverse transcription of packaged cellular RNA species. We have shown that (i) cellular 7S L RNA is an efficient template; (ii) initiation is on a unique tRNA-like primer; (iii) synthesis produces a 135-base strong-stop DNA product; and (iv) after synthesis, RNase H acts to remove the 135 bases of the 7S L RNA which acted as the template. A possible facilitator of such specific transcription may be that, in the virus particles but not in the cell, the majority of the 7S L RNA species exist complexed with the tRNA, even before the disruption of the virus. From the size and sequence features of the reverse transcript of 7S L RNA, we speculate that such events may have participated in the process by which animal cell genomes have, in the course of evolution, accumulated multiple copies of Alu-like elements. PMID- 2985797 TI - Molecular cloning, characterization, and genetic mapping of an endogenous murine mammary tumor virus proviral unit I of C3H/He mice. AB - We have cloned and characterized a novel endogenous murine mammary tumor virus proviral unit of the C3H/He strain of mice. The cloned proviral unit is 16 kilobase pairs (kbp) in size and is composed of a 5.6-kbp 5' EcoRI segment of an endogenous provirus with 10.4-kbp flanking cellular sequences. A comparison of the restriction map of the cloned proviral DNA with that of an endogenous provirus of the GR strain of mice has revealed minor differences in restriction sites on the two proviruses. The restriction enzyme SstI, which does not cleave the 5' EcoRI fragment of GR DNA, cleaves the C3H/He proviral sequences once; MspI has an additional site in the C3H/He proviral sequences. By using a subcloned fragment containing unique cellular sequences as a hybridization probe, we (i) mapped the C3H/He proviral unit to chromosome 14 by using mouse-hamster somatic cell hybrids, and (ii) demonstrated that this proviral unit is also present in the genome of DBA/2 mice. From these results we conclude that the C3H/He strain of mice acquired this proviral unit from DBA stock by genetic transmission. Our data also indicate that the murine mammary tumor virus sequences present in the gag-specific proviral unit of C3H/He mice extend at least 2.45 kbp downstream of the EcoRI site in the genomic DNA. Since the structural organization and chromosomal location of this proviral unit are distinct from those of previously reported proviral units represented by similar-sized (16.7-kbp) EcoRI fragments, we tentatively propose to designate this proviral unit Mtv-7a. PMID- 2985798 TI - High-frequency transduction of c-erbB in avian leukosis virus-induced erythroblastosis. AB - Twenty-one cases of Rous-associated virus type 1-induced erythroblastosis have been analyzed for novel restriction endonuclease fragments of c-erbB. Twenty of the erythroleukemias contained novel c-erbB fragments; 10 of these were found to contain a proviral insertion in c-erbB, and 10 were found to have a new transduction of c-erbB. Each of the proviral insertions was in the same transcriptional orientation as c-erbB, and most appeared to have retained both long terminal repeats as well as 5' viral sequences that signal packaging of RNA into virions. Each of the new c-erbB transducing viruses had a characteristic EcoRI fragment that contained a spliced form of c-erbB sequences. When inoculated into 1-week-old chickens, the new transducing viruses caused rapid-onset erythroblastosis. PMID- 2985799 TI - Induction of angiosarcoma by a c-erbB transducing virus. AB - Recently, 12 new transductions of c-erbB have been identified in a series of Rous associated virus type 1-induced erythroleukemias. During the passage of these new transducing viruses it has become apparent that the erythroleukemia in chicken 5005 contained two different c-erbB transducing viruses. One induces erythroblastosis, whereas the second induces angiosarcoma. The angiosarcoma- and erythroblastosis-inducing viruses appear to have had a common ancestor, since tumors induced by each contain a novel, 4.3-kilobase c-erbB-related EcoRI fragment. The angiosarcoma-inducing virus has been named avian angiosarcoma virus and is designated for the chicken in which it originated. PMID- 2985800 TI - Virion assembly defect of polyomavirus hr-t mutants: underphosphorylation of major capsid protein VP1 before viral DNA encapsidation. AB - The major capsid protein of polyomavirus, VP1, was separated into at least four subspecies by isoelectric focusing. One of these subspecies was selectively extracted from purified virions by mild treatment with sodium dodecyl sulfate, leaving a 140S particle enriched in the other three forms. The two most acidic subspecies were labeled in vivo with [32P]phosphate, and these subspecies are among those identified as being deficient in nontransforming host range (hr-t) mutant virus nonpermissive infection of NIH3T3 cells. Quantitation of VP1 phosphorylation revealed that hr-t mutant virus VP1 is phosphorylated to about 40 to 50% the level of the wild type in NIH3T3 cells, and two-dimensional phosphoamino acid analysis suggested that threonine phosphorylation was affected more than serine phosphorylation. Two results indicate that the VP1 modifications occur before and independent of virus assembly: modified subspecies were detected during wild-type infection within a 2-min pulse-label with [32S]methionine, and VP1 modifications of temperature-sensitive VP1 mutants were the same at both restrictive and permissive temperatures for virus assembly. We conclude that most VP1 modification occurs before viral DNA encapsidation, and that one defect in hr t mutant virus assembly is in VP1 phosphorylation, primarily affecting threonine. PMID- 2985801 TI - An unusual spliced herpes simplex virus type 1 transcript with sequence homology to Epstein-Barr virus DNA. AB - High-resolution transcription mapping localized a spliced 2.7-kilobase herpes simplex virus type 1 mRNA. The 4-kilobase intron of this transcript encodes a nested set of transcripts on the opposite DNA strand. The nucleotide sequence of the DNA encoding the left-hand and right-hand exons of the spliced transcript was determined, and the salient features are presented here. Of major interest is that both exons contained regions within several hundred bases of the splice donor and acceptor sites which showed homology to two regions of the Epstein-Barr virus genome, which are themselves 3 kilobases apart. The spliced herpes simplex virus transcript encoded a translational reading frame which could encode a protein with an approximate size of 75,000 daltons. This value is in agreement with in vitro translation data. The predicted amino acid sequence of the herpes simplex virus protein had significant homology with putative amino acid sequences encoded by the homologous Epstein-Barr virus DNA sequences. PMID- 2985802 TI - Structure of the intracellular defective viral RNAs of defective interfering particles of mouse hepatitis virus. AB - The intracellular defective RNAs generated during high-multiplicity serial passages of mouse hepatitis virus JHM strain on DBT cells were examined. Seven novel species of single-stranded polyadenylic acid-containing defective RNAs were identified from passages 3 through 22. The largest of these RNAs, DIssA (molecular weight [mw], 5.2 X 10(6)), is identical to the genomic RNA packaged in the defective interfering particles produced from these cells. Other RNA species, DIssB1 (mw, 1.9 X 10(6) to 1.6 X 10(6)), DIssB2 (mw, 1.6 X 10(6)), DIssC (mw, 2.8 X 10(6)) DIssD (mw, 0.82 X 10(6)), DIssE (mw, 0.78 X 10(6)), and DIssF (mw, 1.3 X 10(6)) were detected at different passage levels. RNase T1-resistant oligonucleotide fingerprinting demonstrated that all these RNAs were related and had multiple deletions of the genomic sequences. They contained different subsets of the genomic sequences from those of the standard intracellular mRNAs of nondefective mouse hepatitis virus JHM strain. Thus these novel intracellular viral RNAs were identified as defective interfering RNAs of mouse hepatitis virus JHM strain. The synthesis of six of the seven normal mRNA species specific to mouse hepatitis virus JHM strain was completely inhibited when cells were infected with viruses of late-passage levels. However, the synthesis of RNA7 and its product, viral nucleoprotein, was not significantly altered in late passages. The possible mechanism for the generation of defective interfering RNAs was discussed. PMID- 2985803 TI - A single amino acid substitution in a hydrophobic domain causes temperature sensitive cell-surface transport of a mutant viral glycoprotein. AB - DNA sequences were determined for three cDNA clones encoding vesicular stomatitis virus glycoproteins from the tsO45 mutant (which encodes a glycoprotein that exhibits temperature-sensitive cell-surface transport), the wild-type parent strain, and a spontaneous revertant of tsO45. The DNA sequence analysis showed that as many as three amino acid changes could be responsible for the transport defect. By recombining the cDNA clones in vitro and expressing the recombinants in COS cells, we were able to trace the critical lesion in tsO45 to a single substitution of a polar amino acid (serine) for a hydrophobic amino acid (phenylalanine) in a hydrophobic domain. We suggest that this nonconservative substitution may block protein transport by causing protein denaturation at the nonpermissive temperature. Comparison of the predicted glycoprotein sequences from two vesicular stomatitis virus strains suggests a possible basis for the differential carbohydrate requirement in transport of the two glycoproteins. PMID- 2985804 TI - Cells that constitutively express the herpes simplex virus immediate-early protein ICP4 allow efficient activation of viral delayed-early genes in trans. AB - To study the role of herpes simplex virus type 1 immediate-early proteins in the transcriptional activation of herpes simplex virus genes, we isolated stably transformed cells expressing herpes simplex virus type 1 ICP4, an immediate-early protein known from previous studies to be necessary for delayed-early and late transcription. These cells efficiently expressed six delayed-early herpes simplex virus genes introduced by viral superinfection, in the absence of de novo viral protein synthesis. In contrast, the delayed-early gene encoding alkaline exonuclease and the late gene encoding the capsid protein VP5 were expressed at much lower levels. Expression of a second late gene, that for glycoprotein C, was undetectable under the same experimental conditions. These results suggest that many, but not all, delayed-early genes are efficiently activated by ICP4; in addition, they demonstrate that although the late gene for VP5 is detectably activated by ICP4, its full expression requires additional factors. PMID- 2985805 TI - Characterization of the major immediate-early polypeptides encoded by murine cytomegalovirus. AB - The immediate-early (IE) infected cell proteins induced by the murine cytomegalovirus (Smith strain) were studied. These polypeptides were identified as IE proteins by their synthesis in the presence of actinomycin D after removal from a protein synthesis block mediated by cycloheximide. By using a murine antiserum against murine cytomegalovirus, three abundant polypeptides of 89, 84, and 76 kilodaltons (kd) were immunoprecipitated. The three major proteins are phosphorylated but not glycosylated and share antigenic determinants recognized by monoclonal antibodies. The 84 and 76-kd polypeptides represent post translational modification products of the 89-kd protein. Accordingly, in vitro translation of IE infected cell RNA revealed only the 89-kd polypeptide. The viral origin of the RNA species directing the synthesis of the major 89-kd IE polypeptide was verified by hybrid selection of IE RNA with DNA fragments representing the region from 0.769 to 0.815 map units of the murine cytomegalovirus genome. IE polypeptides were found to be located in the nuclei and the cytoplasm of infected cells. Studies on the kinetics of IE polypeptide synthesis revealed negative regulatory effects on IE gene expression correlated with the synthesis of early proteins. PMID- 2985806 TI - Characterization of a variant virus selected in rat brains after infection by coronavirus mouse hepatitis virus JHM. AB - The intracerebral inoculation of Lewis rats with the murine coronavirus MHV-JHM leads in the majority of animals to acute encephalitis and death within 14 days. Viral RNAs isolated from the brains of animals 5 to 7 days after infection were compared by Northern blot analysis with the RNAs produced during the lytic infection of Sac(-) or DBT cells with wild-type MHV-JHM (wt virus). Reproducibly, the subgenomic mRNAs 2 and 3 but no other viral RNAs were significantly larger in the brain-derived material. All viruses isolated from infected brain material displayed and maintained this altered mRNA profile when cultivated in Sac(-) or DBT cells. A virus isolated from the infected brain material, MHV-JHM clone 2 (cl 2 virus), has been further characterized. This isolate grew in tissue culture and induced cytopathic effects comparable to those induced by wt virus. However, the mRNAs 2 and 3 produced in cl-2 virus-infected cells had molecular weights ca. 150,000 larger than those produced in cells infected with wt virus. There was no detectable difference in genome-sized RNA (mRNA 1) or subgenomic mRNAs 4, 5, 6, and 7 as determined by electrophoresis in agarose gels. T1-resistant oligonucleotide analysis of genomic RNA revealed one additional and one missing oligonucleotide in the fingerprint of cl-2 virus compared with wt virus. The oligonucleotide fingerprints of intracellular mRNA 3 were identical for both viruses. Pulse-labeling with [35S]methionine in the presence of tunicamycin showed that the primary translation product of mRNA 3, the E2 apoprotein, was ca. 15,000 larger in molecular weight in cl-2 virus-infected cells. These data show that viruses with larger mRNAs 2 and 3 (the latter encoding an altered E2 glycoprotein) are selected for multiplication in rat brains. Mechanisms for the generation of such variants and the possible nature of their selective advantage are considered. PMID- 2985807 TI - Deletion in the 3' pol sequence correlates with aberration of RNA expression in certain replication-defective avian sarcoma viruses. AB - The RNA expression of a series of replication-defective recovered avian sarcoma viruses (rASVs) were studied. Abnormal-sized viral RNAs, both larger and smaller than the genome, were observed in the nonproducer cells infected with rASVs containing env and pol deletions. Each nonproducer clone contained a single provirus integrated at a unique site and expressed a unique RNA pattern. Upon rescuing of the sarcoma virus with a helper virus and subsequent cloning, the RNA pattern of individual nonproducer clones again displayed variation according to the integration sites. This was not seen in nondefective rASV or in rASVs containing only an env deletion. The aberrant RNA expression did not result from the lack of reverse transcriptase activity per se, since neither nonconditional nor temperature-sensitive mutants of RSV expressed abnormal viral RNAs in the absence of a functional reverse transcriptase. The abnormal RNA patterns could not be corrected in trans by helper virus functions. The unusual-sized RNAs in env- pol- rASV-infected cells are not due to splicing to alternative acceptor sites for src mRNA because there are no extra viral sequences between the 5' leader and the src sequences; instead, they are due to the presence of extra sequences, most likely of cellular origin, at the 3' ends of the viral RNAs. Based upon the extent of deletions in the viral genomes, the data suggest that deletion in the 3' pol region of those rASVs results in a cis effect on the transcription and processing of the 3' ends of viral RNAs. The unusual-sized viral RNAs are most likely due to read-through transcription from the right-hand terminus of provirus into downstream cellular sequences, followed by cleavage and polyadenylation at multiple sites of the 3' region of the RNA transcripts. The extent of read-through transcription appears to depend on the chromosomal location of the provirus. PMID- 2985808 TI - Direct inactivation of viruses by MCP-1 and MCP-2, natural peptide antibiotics from rabbit leukocytes. AB - Six homologous peptides were purified to homogeneity from rabbit granulocytes or alveolar macrophages and tested for their ability to inactivate herpes simplex virus type 1 (HSV-1). Two of the peptides, MCP-1 and MCP-2, showed considerable in vitro neutralizing activity, whereas four structurally homologous peptides (NP 3a, NP-3b, NP-4, and NP-5) were relatively ineffective. Inactivation of HSV-1 by MCP-1 or MCP-2 depended on peptide concentration and on the time, temperature, and pH of the incubation. HSV-2, vesicular stomatitis virus, and influenza virus A/WSN were also susceptible to direct neutralization by MCP-1 or MCP-2, whereas cytomegalovirus, echovirus type 11, and reovirus type 3 were not. We speculate that MCP-1 and MCP-2, peptides that are abundant in rabbit granulocytes and lung macrophages, may contribute to antiviral defenses by mediating the direct inactivation of HSV-1 and selected other viruses. PMID- 2985809 TI - A large-tumor-antigen-specific monoclonal antibody inhibits DNA replication of simian virus 40 minichromosomes in an in vitro elongation system. AB - In productively infected cells, a fraction of large-tumor antigen (T antigen) is tightly bound to replicating simian virus 40 (SV40) minichromosomes and does not dissociate at salt concentrations of greater than 1 M NaCl. We present electronmicrograms demonstrating the presence of T antigen on the replicated sections of replicating SV40 minichromosomes. We also show that the fraction of tightly bound T antigen is recognized by antibodies from mouse tumor serum and, more specifically, by a particular T-antigen-specific monoclonal antibody, PAb 1630. A second T-antigen-specific monoclonal antibody, PAb 101, does not react with the T-antigen fraction remaining on replicating SV40 chromatin at high salt concentrations. We used an in vitro replication system which allows, via semiconservative DNA replication, the completion of in vivo-initiated replicative intermediate DNA molecules. We show that monoclonal antibody PAb 1630, but not monoclonal antibody PAb 101, inhibits viral DNA replication. We discuss the possibility that SV40 T antigen may play a role in chain elongation during SV40 chromatin replication. PMID- 2985810 TI - Simian agent 12 is a BK virus-like papovavirus which replicates in monkey cells. AB - We have begun to characterize the genomic structure and replication of the baboon papovavirus simian agent 12 (SA12). We have defined a wild-type clone of SA12 (SA12 wt100) by plaque purification from a heterogeneous stock. The functional map of SA12 wt100 can be aligned with those of the other primate papovaviruses by assigning one of the two EcoRI sites as 0/1.0 map units. The origin of bidirectional viral DNA replication maps near 0.67 map units, consistent with the limits of sequences homologous to origin sequences in the other papovaviruses. DNA sequence analysis shows that the organization of the SA12 genome is similar to that of the other primate papovaviruses studied. The arrangement and sequence of functional elements in the origin of replication region, as well as the sequences of the N-terminal regions of early protein products, indicate that SA12 is most closely related to the human virus BK, next most closely related to JC virus, and less closely related to simian virus 40. Unlike BK virus, SA12 is capable of productive infection of African green monkey kidney cells. PMID- 2985811 TI - In vitro replication of Sendai virus wild-type and defective interfering particle genome RNAs. AB - A system for studying the in vitro replication of the genome RNAs of Sendai virus and its defective interfering particle DI-H has been developed. Cytoplasmic extracts of baby hamster kidney cells infected with wild-type Sendai virus or coinfected with wild-type Sendai virus plus DI-H were prepared after lysolecithin treatment at 12 h postinfection. The extracts supported the transcription of six viral mRNAs as well as the replication of the Sendai virus 50S (wild-type) and 14S DI-H genome RNAs and their encapsidation into nucleocapsids in the absence of de novo protein synthesis. RNA replication in vitro represented more than 50% of total RNA synthesis, a relative level higher than that found in the infected cell. The proteins required for Sendai virus RNA replication were present in a soluble protein pool at the time of extract preparation. Depletion of the protein pool by prior treatment of infected cells with cycloheximide inhibited subsequent in vitro genome replication without affecting transcription. The cytoplasmic extract may be separated by high-speed centrifugation into two components: the Sendai virus wild-type and DI-H nucleocapsid templates containing the RNA and associated NP, L, and P proteins and the soluble protein fraction containing primarily the P, NP, and M viral proteins with trace amounts of the L, HN, Fo, and nonstructural C proteins. The isolated intracellular DI-H nucleocapsid template alone cannot replicate its RNA, but when recombined with the Sendai virus soluble protein fraction it catalyzes the replication and encapsidation of viral RNAs. The initiation of RNA replication in vitro can be demonstrated because detergent-disrupted purified DI-H virions replicate both positive- and negative-strand RNAs in the presence, but not in the absence, of the soluble protein fraction from an extract of infected cells. PMID- 2985812 TI - Latent and viral replicative transcription in vivo from the BamHI K fragment of Epstein-Barr virus DNA. AB - We mapped one latent and two replicative messages transcribed in vivo from the BamHI K fragment of the Epstein-Barr virus genome. The exon encoding Epstein-Barr nuclear antigen (EBNA), a major latent product, is 2,028 bases; the 3' end of this exon occurs 30 bases after the polyadenylation signal AATAAA, and the 5' end occurs within a splice acceptor site. The open reading frame which encodes the EBNA peptide is completely contained within this coding exon. The exon was faithfully transcribed after transfection of cloned BamHI-K into either COS-1 or TK- mouse L cells. In lymphocytes the abundance of the EBNA message is increased after cycloheximide treatment. The two viral replicative genes completely contained in BamHI-K were not transcribed in line X50-7, in which the genome is tightly latent. In contrast to the EBNA message, these mRNAs of 1.3 and 2.1 kilobases are inducible with phorbol ester and are unspliced. Their promoter regions are similar to those of each other and to replicative promoters mapped in other regions of the Epstein-Barr virus genome (P. J. Farrell, A. Bankier, C. Seguin, P. Deininger, and B. G. Barrell, EMBO J. 2:1331-1338, 1983). An unusual feature of these replicative genes is that the smaller mRNA begins within a long open reading frame of the larger mRNA. The identification of the structure of latent and replicative genes within one DNA fragment will facilitate analysis of regulation of expression for the two life cycles of the virus. PMID- 2985813 TI - Herpes simplex virus 1 reiterated S component sequences (c1) situated between the a sequence and alpha 4 gene are not essential for virus replication. AB - The herpes simplex virus 1 genome consists of two components, L and S, each containing unique sequences flanked by inverted repeats. Each of the 6.5-kilobase pair inverted repeats of the S component, designated a'c' and ca, contains an approximately 700-base pair sequence (designated c1) located between the a sequence and the 3' terminus of the alpha 4 gene. Like the a sequence, c1 consists of direct repeats and unique sequences. Its function is not known. To probe for its function, we constructed a plasmid containing a viral thymidine kinase (TK) gene inserted into the c1 sequence. The construct was recombined into the genome of a TK- virus by cotransfection with intact viral DNA and selection for TK+ virus. As predicted from previous studies (Knipe et al., Proc. Natl. Acad. Sci. U.S.A. 75:3896-3900, 1978), the TK gene was found to be present in both copies of the c1 sequence in the R3104 virus. To delete the c1 sequence we constructed a plasmid containing 4 kilobase pairs of pBR322 flanked by an a sequence and by structural sequences of the alpha 4 gene. In this instance the cells were transfected with the construct and R3104 DNA; the progeny of the transfection was plated in the presence of 5-bromo-2'-deoxyuridine, and the selection was for TK- virus (R3158). The pBR322 DNA sequences replaced the c1 at both termini of the S component in R3158 DNA, but a sequence homologous to c1 was present in proximity to the 3' terminus of the alpha 4 gene. The results indicate that the c1 region has no significant role in the replication of the virus in cell culture. The advantage of inserting the pBR322 sequence is that it permits efficient cloning of large herpes simplex virus 1 DNA fragments by simple ligation of digests and transformation of appropriate Escherichia coli strains. The effortless selection of recombinants carrying inserts in both copies of the c1 restates the usefulness of this technique for selection of insertion deletion recombinants and underscores the rapid emergence of sequence identity at both ends of the reiterated regions of the S component as previously reported (Knipe et al., Proc. Natl. Acad. Sci. U.S.A. 75:3896-3900, 1978). PMID- 2985814 TI - Isolation and structural characterization of cap-binding proteins from poliovirus infected HeLa cells. AB - In poliovirus-infected HeLa cells, poliovirus RNA is translated at times when cellular mRNA translation is strongly inhibited. It is thought that this translational control mechanism is mediated by inactivation of a cap-binding protein complex (comprising polypeptides of 24 [24-kilodalton cap-binding protein], 50, and approximately 220 kilodaltons). This complex can restore the translation of capped mRNAs in extracts from poliovirus-infected cells. We have previously shown that the virally induced defect prevents interaction between cap recognition factors and mRNA. Here, we show that the cap-binding protein complex (and not the 24-kilodalton cap-binding protein) has activity that restores the cap-specific mRNA-protein interaction when added to initiation factors from poliovirus-infected cells. Thus, the activity that restores the cap-specific mRNA protein interaction and that which restores the translation of capped mRNAs in extracts from poliovirus-infected cells, copurify. The results also indicate, by an alternative assay, that the cap-binding protein complex is the only factor inactivated by poliovirus. We also purified cap-binding proteins from uninfected and poliovirus-infected HeLa cells. By various criteria, the 24-kilodalton cap binding protein is not structurally modified as a result of infection. However, the 220-kilodalton polypeptide of the cap-binding protein complex is apparently cleaved by a putative viral (or induced) protease. By in vivo labeling and m7GDP affinity chromatography, we isolated a modified cap-binding protein complex from poliovirus-infected cells, containing proteolytic cleavage fragments of the 220 kilodalton polypeptide. PMID- 2985815 TI - Sequence organization and molecular cloning of mouse mammary tumor virus DNA endogenous to C57BL/6 mice. AB - The sequence organization of mouse mammary tumor virus DNA endogenous to the C57BL/6 inbred mouse strain was characterized by Southern blot analysis, utilizing probes specific for particular regions of the mouse mammary tumor virus provirus and by molecular cloning of endogenous mouse mammary tumor virus DNA. The genome of C57BL/6 mice contains three apparently intact, endogenous proviral units; two of these units comprise the Mtv-8 (unit II) and Mtv-9 (unit III) genetic loci that are also present in the DNA of BALB/c mice. The third unit is defined by EcoRI restriction fragments of 10.0 and 8.4 kilobases that contain the 5' and 3' portions of the provirus, respectively. This unit, termed unit XI and encoded by the genetic locus Mtv-17, has not been previously recognized in C57BL/6 DNA, but it can be clearly distinguished from the proviral units at Mtv-8 and Mtv-9 by Southern blot analysis under appropriate conditions. The proviral unit at Mtv-17 is not present in BALB/c DNA. DNAs comprising the entire Mtv-8 locus and the 3' portions of Mtv-9 and Mtv-17 were cloned. Analysis of the cloned DNA revealed no obvious deletions or rearrangements that would render proviral DNA defective; however, these endogenous genes are normally not transcriptionally active. PMID- 2985816 TI - Simian virus 40 and polyomavirus large tumor antigens have different requirements for high-affinity sequence-specific DNA binding. AB - By using a DNA fragment immunoassay, the binding of simian virus 40 (SV40) and polyomavirus (Py) large tumor (T) antigens to regulatory regions at both viral origins of replication was examined. Although both Py T antigen and SV40 T antigen bind to multiple discrete regions on their proper origins and the reciprocal origin, several striking differences were observed. Py T antigen bound efficiently to three regions on Py DNA centered around an MboII site at nucleotide 45 (region A), a BglI site at nucleotide 92 (region B), and another MboII site at nucleotide 132 (region C). Region A is adjacent to the viral replication origin, and region C coincides with the major early mRNA cap site. Weak binding by Py T antigen to the origin palindrome centered at nucleotide 3 also was observed. SV40 T antigen binds strongly to Py regions A and B but only weakly to region C. This weak binding on region C was surprising because this region contains four tandem repeats of GPuGGC, the canonical pentanucleotide sequence thought to be involved in specific binding by T antigens. On SV40 DNA, SV40 T antigen displayed its characteristic hierarchy of affinities, binding most efficiently to site 1 and less efficiently to site 2. Binding to site 3 was undetectable under these conditions. In contrast, Py T antigen, despite an overall relative reduction of affinity for SV40 DNA, binds equally to fragments containing each of the three SV40 binding sites. Py T antigen, but not SV40 T antigen, also bound specifically to a region of human Alu DNA which bears a remarkable homology to SV40 site 1. However, both tumor antigens fail to precipitate DNA from the same region which has two direct repeats of GAGGC. These results indicate that despite similarities in protein structure and DNA sequence, requirements of the two T antigens for pentanucleotide configuration and neighboring sequence environment are different. PMID- 2985817 TI - Mechanisms of monoclonal antibody-mediated protection against virulent Semliki Forest virus. AB - Both neutralizing and nonneutralizing immunoglobulin G2a monoclonal antibodies (MAs) directed against the E2 glycoprotein of Semliki Forest virus (SFV) protected mice prophylactically and therapeutically against virulent SFV infection. The neutralizing MAs, however, conferred protection to mice at lower doses than did nonneutralizing MAs. The antibody-dependent, complement-mediated cytolysis of SFV-infected L cells was effectuated by both kinds of antibodies, but again neutralizing MAs were more effective. Removal of the Fc part of the neutralizing MA UM 5.1 by pepsin digestion resulted in a 100-fold reduction of the neutralization titer (10(4) versus 10(6)) and a complete loss of its capacity to mediate antibody-dependent, complement-mediated cytolysis. Passive protection of infected mice occurred only after administration of relatively high doses of F(ab')2 of MA UM 5.1 (30.0 micrograms versus 0.1 microgram). F(ab')2 fragments prepared from the nonneutralizing MA UM 4.2 had lost their protective capacity completely. Surprisingly, the nonneutralizing MA UM 4.2 retarded virus growth in mouse fibroblasts (L cells), although inhibition was at much higher doses than with the neutralizing MA UM 5.1. Furthermore, both MAs promoted the uptake of virulent SFV in the Fc receptor-bearing WEHI-3 cells. The results suggest that nonneutralizing MAs protect mice not only by antibody-dependent, complement mediated cytolysis but also by growth inhibition and enhanced uptake of SFV in the nonpermissive macrophages of BALB/c mice. This hypothesis is supported by the absence of viremia in recipients of nonneutralizing MA UM 4.2 at 24 h after infection. PMID- 2985818 TI - Purification of Epstein-Barr virus DNA polymerase from P3HR-1 cells. AB - The Epstein-Barr virus DNA polymerase was purified from extracts of P3HR-1 cells treated with n-butyrate for induction of the viral cycle. Sequential chromatography on DNA cellulose, phosphocellulose, and blue Sepharose yielded an enzyme preparation purified more than 1,300-fold. The purified enzyme was distinct from cellular enzymes but resembled the viral DNA polymerase in cells infected with herpes simplex virus type 1 or 2. The active enzyme had an apparent molecular weight of 185,000 as estimated by gel filtration on Sephacryl S-300. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major polypeptide corresponding to a molecular weight of ca. 110,000. This polypeptide correlated with the catalytic function of the purified enzyme, whereas the other, less abundant polypeptides did not. By immunoblotting, the 110,000-molecular weight polypeptide could be identified as a viral polypeptide. It could not be determined whether the native enzyme was composed of more than one polypeptide. PMID- 2985819 TI - Mutations near the carboxyl terminus of the simian virus 40 large tumor antigen alter viral host range. AB - We report the characterization of three mutants of simian virus 40 with mutations that delete sequences near the 3' end of the gene encoding large tumor antigen (T antigen). Two of these mutants, dl1066 and dl1140, exhibit an altered viral host range. Wild-type simian virus 40 is capable of undergoing a complete productive infection on several types of established African green monkey kidney lines, including BSC40 and CV1P. dl1066 and dl1140 grow on BSC40 cells at 37 degrees C. However, both mutants fail to form plaques on BSC40 cells at 32 degrees C or on CV1P cells at any temperature. These mutants are capable of replicating viral DNA in the nonpermissive cell type, indicating a defect in an activity of T antigen not related to its replication function. Furthermore this defect can be complemented in trans by the wild type or by a variety of DNA replication negative T antigen mutants, so long as they produce a normal carboxyl-terminal region of the molecule. Our data are consistent with the hypothesis that the C terminal region of T antigen constitutes a functional domain. We propose that this domain encodes an activity that is required for simian virus 40 productive infection on the CV1P cell line, but not on BSC40. PMID- 2985820 TI - Molecular cloning of proviral DNA and structural analysis of the transduced myc oncogene of avian oncovirus CMII. AB - Molecularly cloned proviral DNA of avian oncogenic retrovirus CMII was isolated by screening a genomic library of a CMII-transformed quail cell line with a myc specific probe. On a 10.4-kilobase EcoRI fragment, the cloned DNA contained 4.4 kilobases of CMII proviral sequences extending from the 5' long terminal repeat to the EcoRI site within the partial (delta) complement of the env gene. The gene order of CMII proviral DNA is 5'-delta gag-v-myc-delta pol-delta env-3'. All three structural genes are partially deleted: the gag gene at the 3' end, the env gene at the 5' end, and the pol gene at both ends. The delta gag (0.83 kilobases) v-myc (1.50 kilobases) sequences encode the p90gag-myc transforming protein of CMII. In comparison with the p110gag-myc protein of acute leukemia virus MC29, p90gag-myc lacks amino acids corresponding to additional 516 bases of gag sequences and 12 bases of 5' v-myc sequences present in the MC29 genome. Nucleotide sequence analysis of CMII proviral DNA at the delta gag-v-myc and the v-myc-delta pol junctions revealed significant homologies between avian retroviral structural genes and the cellular oncogene c-myc precisely at the positions corresponding to the gene junctions in CMII. Furthermore, the delta gag v-myc junction in CMII corresponds to sequence elements in gag and C-myc that are possible splicing signals. The data suggest that transduction of cellular oncogenes may involve RNA splicing and recombination with homologous sequences on retroviral vectors. Different sequence elements of both the retroviral vectors and the c-myc gene recombined during genesis of highly oncogenic retroviruses CMII, MC29, or MH2. PMID- 2985821 TI - Development and analysis of a transformation-defective mutant of Harvey murine sarcoma tk virus and its gene product. AB - The Harvey murine sarcoma virus has been cloned and induces focus formation on NIH 3T3 cells. Recombinants of this virus have been constructed which include the thymidine kinase gene of herpes simplex virus type 1 in a downstream linkage with the p21 ras gene of Harvey murine sarcoma virus. Harvey murine sarcoma tk virus rescued from cells transfected with this construct is both thymidine kinase positive and focus inducing in in vitro transmission studies. The hypoxanthine aminopterin-thymidine selectability of the thymidine kinase gene carried by this virus has been exploited to develop three mutants defective in the p21 ras sequence. All three are focus negative and thymidine kinase positive when transmitted to suitable cells. Of these, only one encodes a p22 that is immunologically related to p21. This mutant has been used to explore the relationship between the known characteristics of p21 and cellular transformation. Data presented herein indicate that the p21 of Harvey murine sarcoma virus consists of at least two domains, one which specifies the guanine nucleotide-binding activity of p21 and the other which is involved in p21 membrane association in transformed cells. PMID- 2985822 TI - Mass and molecular composition of vesicular stomatitis virus: a scanning transmission electron microscopy analysis. AB - Dark-field scanning transmission electron microscopy was used to perform mass analyses of purified vesicular stomatitis virions, pronase-treated virions, and nucleocapsids, leading to a complete self-consistent account of the molecular composition of vesicular stomatitis virus. The masses obtained were 265.6 +/- 13.3 megadaltons (MDa) for the native virion, 197.5 +/- 8.4 MDa for the pronase treated virion, and 69.4 +/- 4.9 MDa for the nucleocapsid. The reduction in mass effected by pronase treatment, which corresponds to excision of the external domains (spikes) of G protein, leads to an average of 1,205 molecules of G protein per virion. The nucleocapsid mass, after compensation for the RNA (3.7 MDa) and residual amounts of other proteins, yielded a complement of 1,258 copies of N protein. Calibration of the amounts of M, NS, and L proteins relative to N protein by biochemical quantitation yielded values of 1,826, 466, and 50 molecules, respectively, per virion. Assuming that the remaining virion mass is contributed by lipids in the viral envelope, we obtained a value of 56.1 MDa for its lipid content. In addition, four different electron microscopy procedures were applied to determine the nucleocapsid length, which we conclude to be 3.5 to 3.7 micron. The nucleocapsid comprises a strand of repeating units which have a center-to-center spacing of 3.3 nm as measured along the middle of the strand. We show that these repeating units represent monomers of N protein, each of which is associated with 9 +/- 1 bases of single-stranded RNA. From scanning transmission electron microscopy images of negatively stained nucleocapsids, we inferred that N protein has a wedge-shaped, bilobed structure with dimensions of approximately 9.0 nm (length), approximately 5.0 nm (depth), and approximately 3.3 nm (width, at the midpoint of its long axis). In the coiled configuration of the in situ nucleocapsid, the long axis of N protein is directed radially, and its depth corresponds to the pitch of the nucleocapsid helix. PMID- 2985823 TI - Construction and use of cDNA clones for the mapping and identification of Epstein Barr virus early P3HR-1 mRNAs. AB - cDNA clones, specific for early Epstein-Barr virus (EBV) RNAs, were constructed from total cytoplasmic RNA of P3HR-1 TK- cells. From 10,000 cDNA clones screened, 22 virus-specific cDNA clones were selected by hybridization with a total EBV DNA. These clones were then precisely mapped on the EBV genome and the corresponding mRNAs were identified by Northern blot hybridizations. Most of them are clearly related to some of the open reading frames described by Baer et al. (Nature [London] 310:207-211, 1984). They represent at least 18 different genes active during the early viral cycle. The transcriptional activity of the virus during the early stage was also studied by dot blot hybridization of total early cDNA probe to EBV genomic fragments. Three main regions showed very strong hybridization with the cDNA probe: BamHI a, M, and L fragments, BamHI K, B, and G fragments, and BamHI B1 fragment (deleted in strain B95-8) and the adjacent right end of the DNA molecule. Seventeen of the cDNA clones were localized in these highly transcribed regions. The five others were dispersed all along the EBV genome. PMID- 2985824 TI - A genetic test for expression of a functional herpes simplex virus DNA-binding protein from a transfected plasmid. AB - The major DNA-binding protein, ICP8, encoded by herpes simplex virus is localized to the infected cell nucleus where it plays a role in viral DNA replication and control of viral gene expression. To identify the parts of the ICP8 protein that are important for its localization and functions, we have developed a system to test the ability of recombinant plasmids to express functional ICP8. A recombinant plasmid containing the wild-type ICP8 gene was transfected into cells. The cells were later infected with a temperature-sensitive ICP8 mutant virus at the nonpermissive temperature. Sufficient wild-type ICP8 was expressed from the transfected plasmid to complement the replication of the mutant virus. This provides a genetic system to test the properties of ICP8 expressed from mutagenized plasmids without the establishment of a stable cell line or the reintroduction of the ICP8 gene into the herpes simplex virus genome. PMID- 2985825 TI - A candidate for a new serotype of human rotavirus. AB - We investigated genetic and serological characteristics of a human rotavirus isolate from Indonesia which had a "super short" RNA electrophoretic pattern (A. Hasegawa, S. Inouye, S. Matsuno, K. Yamaoka, R. Eko, and W. Suharyono, Microbiol. Immunol. 28:719-722, 1984). This virus, strain 69M, was found by RNA-RNA hybridization to have a low degree of homology with the representative strains of all four human serotypes. Furthermore, it could not be classified by neutralization analysis into any of these serotypes. Therefore, this virus might belong to a new serotype. PMID- 2985826 TI - The pX region of the bovine leukemia virus is transcribed as a 2.1-kilobase mRNA. AB - The bovine leukemia virus mRNAs expressed in cultured bovine cells of various origins are a 9.0-kilobase genomic RNA, a 5.1-kilobase env RNA, and a newly detected 2.1-kilobase RNA corresponding to the transcription of pX sequences located in between the env gene and the 3' end of the provirus. PMID- 2985827 TI - Human rhinovirus 14 infection of HeLa cells results in the proteolytic cleavage of the p220 cap-binding complex subunit and inactivates globin mRNA translation in vitro. AB - One of the characteristics of picornavirus infection of cells in tissue culture is a specific inhibition of utilization of host cell mRNA for protein synthesis. In this study we show that human rhinovirus 14 is similar to poliovirus in that the inhibition of host cell translation that occurs during infection correlates with the proteolytic cleavage of an Mr 220,000 subunit of the cap-binding protein complex. PMID- 2985828 TI - Fine mapping and sequencing of a variable segment in the inverted repeat region of varicella-zoster virus DNA. AB - A strain variation in the internal and terminal repeats which bind the short unique sequence of varicella-zoster virus (VZV) DNA was found to be due to an insertion or deletion of DNA sequences at a single site. DNA sequence analysis showed that the nucleotide sequence CCGCCGATGGGGAGGGGGCGCGGTACC is tandemly duplicated a variable number of times in different VZV strains and is responsible for the observed variation in mobilities of restriction fragments from this region of VZV DNA. The variable region sequence shares some homology with tandemly repeated regions in the a and c sequences of herpes simplex virus type 1 and probably exists in a noncoding region of the VZV genome. PMID- 2985829 TI - Shutoff of host translation by encephalomyocarditis virus infection does not involve cleavage of the eucaryotic initiation factor 4F polypeptide that accompanies poliovirus infection. AB - Studies were conducted to determine whether encephalomyocarditis virus infection causes proteolytic cleavage of any of the polypeptides which comprise eucaryotic initiation factor 4F. Since no such alterations in the components of the initiation factor were detected, these observations confirmed that the mechanisms whereby encephalomyocarditis virus and poliovirus shut off host translation are different. PMID- 2985831 TI - A clinicopathological and virological study of interstitial cystitis. AB - We studied 41 patients with chronic interstitial cystitis. Histological examination of bladder lesions revealed mucosal ulceration, pancystitis and perineural inflammatory infiltrates. Perineural cell infiltration is related probably to the characteristic symptoms of the disease. A search for a viral etiology, particularly herpes simplex virus, rendered negative results. PMID- 2985830 TI - DNA sequence alterations responsible for the synthesis of thermosensitive VP1 in temperature-sensitive BC mutants of simian virus 40. AB - The segment of simian virus 40 (SV40) genome which is recognized as the BC domain encodes for the COOH-terminal end of the SV40 major capsid protein VP1. Mutations in this domain lead to the synthesis of a thermosensitive VP1 which fails to assemble mature SV40 at the nonpermissive temperature. We determined the DNA sequences of eight BC mutants and compared them with the DNA sequences of wild type SV40, polyomavirus, and BK virus. We found that BC11 and BC223 mutations result from changes in nucleotide residues 2367 (A to C) and 2084 (C to T), respectively. The others (i.e., BC208, BC214, BC216, BC217, BC248, and BC274) share the same point mutation at nucleotide 2354 (C to T). These mutations resulted in the following changes: Lys to Thr, His to Tyr, and Pro to Ser at VP1 amino acid residues 290, 196, and 286, respectively. PMID- 2985832 TI - Malignant mixed mesodermal tumor of bladder occurring after radiotherapy for cervical cancer: report of a case. AB - A rare case of radiation-induced malignant mixed mesodermal tumor of the bladder is reported. A 78-year-old woman complained of hematuria, which originated from a polypoid tumor in the bladder 15 years after radiotherapy for squamous cell carcinoma of the uterine cervix. The bladder tumor recurred 9 months after resection and partial cystectomy then was performed. Histological findings revealed malignant mixed mesodermal tumor composed of carcinomatous and sarcomatous elements. The former element consisted mainly of transitional cell carcinoma with occasional foci of squamous metaplasia and glandular differentiation, while the latter consisted of spindle cell sarcoma showing partly cartilaginous and osseous differentiation. The histogenesis of malignant mixed mesodermal tumor of the bladder also is discussed. PMID- 2985833 TI - Potassium bicarbonate supplementation blocks the hypocalciuric response to hydrochlorothiazide in rats. AB - Bicarbonate salts and thiazide diuretics each lower urinary calcium excretion. This study was undertaken to compare the effects of administering oral sodium or potassium bicarbonate supplements (4.5 mmol./day of each salt) each with or without hydrochlorothiazide (10 mg./kg. body weight/day) on urinary calcium excretion in rats. Urinary calcium decreased (p less than 0.01) by about 50 per cent after sodium bicarbonate supplementation and potassium bicarbonate supplementation. The hypocalciuric response to each bicarbonate salt was similar. However, although hydrochlorothiazide depressed urinary calcium in rats consuming sodium bicarbonate, rats receiving equimolar supplements of potassium bicarbonate did not lower urinary calcium when given hydrochlorothiazide, despite evidence of the expected thiazide-mediated diuresis. It is concluded that in the rat potassium bicarbonate loading blocks the ability of hydrochlorothiazide to lower urinary calcium excretion. PMID- 2985835 TI - Is AIDS caused by a neurotropic virus? PMID- 2985834 TI - Travelers' diarrhea. NIH Consensus Development Conference. AB - Diarrhea is the major health problem in travelers to developing countries. Travel to high-risk areas in Latin America, Africa, the Middle East, and Asia is associated with diarrhea rates of 20% to 50%. The syndrome is caused by an infection acquired by ingesting fecally contaminated food or beverages. Escherichia coli, a common species of enteric bacteria, is the leading pathogen, although a host of other bacteria, viruses, and protozoa have been implicated in some cases. Prudent dietary and hygienic practices should be followed, and they will prevent some, but not all, diarrhea. Antimicrobial agents are not recommended for prevention of TD. Such widespread usage in millions of travelers would cause many side effects, including some severe ones, while preventing a disease that has had no reported mortality. Instead of universal antimicrobial prophylaxis, a more sensible approach is rapid institution of effective treatment that can shorten the disease to 30 hours or less in most people. For mild diarrhea, an antimotility drug such as diphenoxylate or loperamide could be taken. Alternatively, bismuth subsalicylate, which works somewhat slower, can be used. For more severe diarrhea, an antimicrobial drug may be used for treatment, and trimethoprim-sulfamethoxazole, trimethoprim alone, and doxycycline are among the choices. These drugs could be carried by the traveler for use in the event of illness. Oral rehydration should be instituted when necessary. The millions of Americans who travel annually to developing countries and their physicians must be warned of the potential risks of prophylactic antimicrobial drugs, with the attendant side effects in otherwise healthy individuals, and should be informed of the alternative method of prompt, effective treatment of diarrhea. PMID- 2985836 TI - A cluster of young patients with Burkitt's lymphoma and nasopharyngeal carcinoma in central Texas. AB - During a five-month period in 1982, diagnoses were made of Burkitt's lymphoma in three children and nasopharyngeal carcinoma in three teenagers who were residents of a small geographic area of central Texas. The observed number of these two Epstein-Barr virus-related cancers was significantly greater than the number expected in young patients based on Surveillance, Epidemiology, and End Results incidence rates. No epidemiologic associations were found linking any of the patients. Environmental studies including serological assays to determine a specific viral etiology were unrevealing. A search for new agents and the preservation of materials for future studies should continue to be a high priority in the evaluation of cancer clusters. PMID- 2985837 TI - [A clinical study of radiotherapy with HCFU for advanced and recurrent breast cancers. Radiation and HCFU Study Group for breast cancer]. AB - We have investigated the usefulness of combination therapy with radiation and HCFU for advanced and recurrent breast cancer according to a clinical controlled multicenter trial from 1982 to 1984. One hundred cases were registered and 82 of them were available. Treatment sites were the lymph nodes, skin, bone and lung, and the overall response rate was 58% in CR and 19% in PR, while the duration of remission was 18 weeks in CR. Side effects were found in 10% of the patients. Combination therapy with radiation and HCFU may be useful in multimodal treatment for advanced recurrent breast cancer. PMID- 2985838 TI - [Percutaneous fine needle biopsy for malignant mediastinal tumors]. AB - Seventeen patients with mediastinal malignant tumors (four neurogenic tumors, four thymomas, three germ cell tumors, two malignant lymphomas and one each of carcinoid, malignant melanoma, multiple myeloma and carcinoma simplex) were examined for the evaluation of percutaneous fine needle biopsy. In 16 cases of them, adequate materials for cytodiagnosis were obtained. Fifteen of the 17 cases (88%) were positive for malignant cells, and 1 case was negative. As for the histological types, 9 cytological diagnoses (60%) were compatible and 4 diagnoses (27%) were not. Regarding complications of percutaneous fine needle biopsy, 1 case of subcutaneous needle tract implantation was noted. Percutaneous fine needle biopsy is strongly recommended for diagnosis of mediastinal tumors. PMID- 2985839 TI - [Malignant fibrous histiocytoma of the thoracic region]. AB - A clinicopathological investigation of five malignant fibrous histiocytoma (MFH) in the thoracic region was performed. They were two cases of metastasis to the lung and 1 each of the primary lesion in the lung, chest wall and sternal region. This tumor usually occurs on the extremities, in the abdominal cavity or the retroperitoneal areas, so instances of its arising in the thoracic region, except for metastatic episodes, are considered to be rare. Tumors in 4 cases were excised surgically, while one was inoperable. On light microscopy, the lesions were found to be composed of two types of neoplastic cells, fibroblast-like cells and histiocyte-like cells, showing the characteristic histologic pattern of storiform and pleomorphic. Electron-microscopically, these two types of cells were discriminated according to the quantitative and quantitative differences of the various intracytoplasmic organelles. The histogenesis of malignant fibrous histiocytoma has been commonly considered to be the histiocytic cell derived from the bone marrow. However, more detailed pathological investigations are necessary. PMID- 2985840 TI - [Adenocarcinoma of the esophagus (early stage) concomitant with lung cancer--a case report]. AB - A case of early esophageal cancer, 2.5 cm in length, located just above the esophagogastric junction is presented. This cancer seemed to originate from the fundic gland of the esophagus, because it consisted of the submucosal invasion of well-differentiated adenocarcinoma which was almost completely covered with the squamous cell mucosa of the esophagus. This patient was referred to our clinic because of lower to be a small-cell carcinoma of the lung extending to the mediastinum. He died of the lung cancer one year after operation. PMID- 2985842 TI - [Clinicopathological studies of chronic graft rejection]. PMID- 2985841 TI - [A case report of a double cancer of the ureter and stomach]. AB - A 63-year-old man complained of hunger epigastralgia. X-ray and endoscopic examination demonstrated Borrmann II type gastric carcinoma. Radical gastrectomy was performed on October 20, 1982. The histological findings revealed moderately differentiated tubular adenocarcinoma. In May 1983, right hydronephrosis was recognized by ultrasonography. A papillary tumor was visualized in the right ureter in retrograde pyelography, and percutaneous transrenal urinary tract drainage was performed. Cytological findings revealed class V. Total nephro uretectomy was performed on June 16, 1983. The histological findings revealed transitional cell carcinoma. PMID- 2985843 TI - [Maintenance hemodialysis. IV. Complications in dialysis patients and their management]. PMID- 2985844 TI - [A case of minute hepatocellular carcinoma with calcification]. PMID- 2985845 TI - [CT findings of metastatic ovarian tumors]. PMID- 2985846 TI - [Results of the treatment of small cell lung cancer by radiation or radiation and chemotherapy]. PMID- 2985848 TI - [A case of WDHA syndrome and the studies on 14 domestic cases]. PMID- 2985847 TI - [A case report of resected hepatocellular carcinoma after renal transplantation in a HBsAg positive patient]. PMID- 2985849 TI - [Plasma PIVKA-II levels in various liver diseases--with special reference to plasma PIVKA-II as a marker of hepatocellular carcinoma]. PMID- 2985850 TI - [Measurement of the change of the portal blood flow volume after TAE therapy by ultrasonic pulsed Doppler method]. PMID- 2985851 TI - [An epidemiological study on relationship between intake of dietary fiber and colonic diseases]. PMID- 2985852 TI - Effects of calcium hopantenate on neurotransmitter receptors in the rat brain. AB - Effects of calcium hopantenate (HOPA) on neurotransmitter and neuropeptide receptors in the central nervous system (CNS) were investigated. In the radioreceptor assay (RRA), HOPA inhibited the [3H]-gamma-aminobutylic acid (GABA) receptor binding in a dose-dependent manner with a cross-reactive potency of 0.2%. On the other hand, radiolabeled ligand binding to CNS receptors in the benzodiazepine (BDZ)-, muscarinic cholinergic (mACh)-, methionine-enkephalin (ENK)- and thyrotropin releasing hormone (TRH)-RRA systems was not inhibited even by the addition of HOPA up to 100 microM. Repeated injection of HOPA (250 mg/kg/day for 7 consecutive days) increased GABA receptor binding by 53% in the cerebral cortex, while GABA binding in the rest of the forebrain did not change. The increased GABA receptor binding in the cerebral cortex of HOPA treated rats was due to the increased affinity of the binding sites. BDZ-, mACh-, ENK- and TRH receptor bindings were not affected in either the cerebral cortex or the rest of the forebrain by repeated injection of HOPA. These results suggest that at least a part of the therapeutic efficacy of HOPA is due to sensitization of the GABA receptor in the cerebral cortex. PMID- 2985853 TI - Serum CEA levels facilitate detection of recurrences of cancer in patients after gastrectomy. AB - In an attempt to assess the usefulness of carcinoembryonic antigen (CEA) for predicting the progression of gastric cancer, CEA productivity was evaluated according to serum CEA levels, at the time of recurrence or relapse. In cases of a recurrence, abnormal CEA levels were observed in 14 of 17 (82.4 per cent) with differentiated carcinoma and in 9 of 21 (42.9 per cent) with undifferentiated carcinoma. Preoperative abnormal CEA levels were observed in only 6 of 41 patients (14.6 per cent). A median lead time of manifestation of recurrence was 5 months. In those with relapse, 9 of 11 (81.8 per cent) patients with differentiated carcinoma and 13 of 18 (72.2 per cent) with an undifferentiated carcinoma had abnormal CEA levels. Preoperative abnormal CEA levels were observed in 24 of these 89 patients (27.0 per cent). Postoperative monitoring of CEA seems to be useful for early recognition of gastric cancer progression, irrespective of the preoperative CEA levels. PMID- 2985854 TI - Streptozotocin effective for treating multiple-hormone-producing malignant islet cell tumor. AB - A woman with a multiple-hormone-producing pancreatic islet cell tumor with hepatic metastases and with recurrent hypoglycemic attacks, was treated with streptozotocin. After this treatment, the elevated serum levels of insulin, C peptide, glucagon and serotonin fell markedly and the low level of fasting blood glucose returned to normal. In accordance with these hormonal changes, scintiscan and CT scan revealed marked regression of the metastatic tumors in the liver. She is alive at this writing, five years after the streptozotocin treatment. Streptozotocin should thus be considered for treatment of malignant islet cell carcinoma with liver metastases and which is not amenable to surgery. PMID- 2985855 TI - Antitumor effect of lipiodolized doxorubicin on VX2 carcinoma in rabbits. AB - Twelve-day-old VX2 carcinoma was inoculated in the hind leg of 24 rabbits and, after 12 days, Doxorubicin (Adriamycin) suspended in Lipiodol Ultra-Fluid (Lipiodol) was then given through the femoral artery. A selective deposit of the contrast material in the tumor for an extended time was evident on the x-rays and the antitumorous effect was remarkable. Lipiodolized antitumor agents warrant further investigation for possible clinical application. PMID- 2985856 TI - Hormone dependency of human breast neoplasms cultured in vitro in the stem cell assay. AB - The stem cell soft agar culture system provides a biological tool with which to study tumor clones derived from heterogeneous tumor cell populations. For testing the feasibility of this approach in identifying hormone-responsive clones of mammary tumors, a study was done to determine whether estrogen deprivation or supplementation would alter colony formation in sequential breast neoplasms received in the Cell Kinetics Laboratory, The Pennsylvania State University. Single-cell suspensions obtained from 22 freshly excised breast neoplasms (20 malignant, 2 benign) were plated in soft agar with and without tamoxifen (Tam) (10(-7) M) with the use of serum-supplemented media and with and without 17 beta estradiol (E2) (10(-8) M] under serum-free medium conditions. Twenty tumors successfully grew, producing 42 +/- 8 (mean +/- SEM) colonies in control dishes in the presence of serum and 27 +/- 5 in its absence (P less than .001). Tam inhibited colony formation in 78% of malignant tumors, whereas a stimulatory effect was observed with E2 in 94%. An increase in colony formation was induced by Tam in 4 tumors (2 benign and 2 malignant). The effects of E2 and Tam on tumor growth were not influenced by the estrogen and progesterone receptor status of the tumor. These preliminary results suggest that all tumors, irrespective of their receptor status, may contain clones of hormone-responsive cells. PMID- 2985857 TI - Influence of antibody infusion on pathogenesis of experimental feline leukemia virus infection. AB - For examination of the influence of antibody on the pathogenesis of feline leukemia virus (FeLV) infection, 12 weanling specific-pathogen-free cats were inoculated with isolates of FeLV and were treated beginning at 7, 19, 21, 24, 34, or 49 days post inoculation (DPI) with feline anti-FeLV hyperimmune serum (10 infusions, 37 mg globulin/kg each at 48-hr intervals). Anti-FeLV serum infusion initiated at 7 DPI prevented the onset of hematopoietic cell infection and viremia. Antibody treatment initiated at 19 or 24 DPI abrogated recently established FeLV viremia and extinguished p27 expression in bone marrow and blood cells. Viremia established for longer periods was refractory to antibody infusion despite establishment of enzyme-linked immunosorbent assay antibody titers of 1:80 to 1:320 in the treated cats. Latent FeLV infection was a sequel to antibody induced curtailment of viral replication in bone marrow cells and was able to reactivate spontaneously in vivo as well as in vitro. PMID- 2985858 TI - Hepatocarcinogenicity of benzo[a]pyrene to rainbow trout by dietary exposure and intraperitoneal injection. AB - The influence of benzo[a]pyrene [(BP) CAS: 50-32-8] on the induction of certain enzymes within the hepatic mixed-function oxidase (MFO) system and its potential carcinogenicity were examined in rainbow trout (Salmo gairdneri). Nine-week feeding trials were performed with 500 and 1,000 ppm BP to determine trout tolerance to BP. Levels of MFO enzymes, including ethoxyresorufin-O-deethylase (EROD), ethoxycoumarin-O-deethylase (ECOD), benzo[a]pyrene monooxygenase (BPMO), and cytochrome P450 were measured during this time. An 18-month feeding trial of a 1,000-ppm BP dose was initiated in duplicate groups of 100 fingerling rainbow trout. Samples of trout were killed at 6, 12, and 18 months for gross and histologic examination of the internal organs for neoplasms. A group of fifty 10 month-old rainbow trout were given 12 monthly ip injections of 1 mg BP in 0.4 ml propylene glycol (PG), and comparable controls were given PG injections only. The trout were held for an additional 6 months, killed at age 28 months, and examined as in the dietary study. Mean MFO enzyme levels of EROD, ECOD, BPMO, and cytochrome P450 were significantly (P less than .001) elevated, showing dose- and time-response relationships when compared to MFO enzyme levels in control fish. Twelve months after BP exposure was initiated, 15% of the BP-fed fish had histologically confirmed neoplasms of the liver. After 18 months the incidence increased to 25%. No evidence of neoplasia was observed in control fish. BP injected ip resulted in a 50% incidence of hepatocellular neoplasms and in a fibrosarcoma of the liver and papillary adenomas of the swim bladder in 1 fish. These results indicate that BP is a potent inducer of selected hepatic MFO enzymes and establish, for the first time, the hepatocarcinogenicity of BP in an aquatic species. PMID- 2985859 TI - No expression of a Rous sarcoma virus-induced tumor antigen in mammalian cells infected with retroviruses transducing other oncogenes of the src gene family. AB - Immunization with mouse and rat cells transformed by Rous sarcoma virus (RSV) or by B77 avian sarcoma virus (ASV) induced complete transplantation resistance against an RSV-induced mouse tumor (CSA1M) in syngeneic hosts. In contrast, most of the mice immunized with a Fujinami sarcoma virus-transformed rat fibroblast line (FSV-3Y1), a feline sarcoma virus-transformed cat fibroblast line (FeSV FEF), an Abelson leukemia virus-infected Balb/3T3 cell line (AbLV-3T3), or an uninfected 3Y1 cell line could not reject the CSA1M. Serologic analysis with the use of a complement-dependent cytotoxicity assay supported the results of transplantation studies. The mouse and rat cells transformed by RSV or B77 ASV expressed a common tumor-specific cell surface antigen (TSSA) detected by syngeneic antiserum against the CSA1M, whereas none of the FSV-3Y1, FeSV-FEF, and AbLV-3T3 cells expressed the TSSA. These results suggest that the common TSSA in the mouse and rat cells transformed by RSV or B77 ASV containing src gene is not shared with mammalian cells infected with retroviruses transducing other oncogenes of the src gene family (i.e., fps, fes, and abl). PMID- 2985860 TI - [Multiple primary cancer of the large intestine associated with chemical burn of the rectum]. PMID- 2985861 TI - [Herpes simplex infection in the newborn infant]. AB - A case of neonatal Herpes simplex infection is being described, which was diagnosed clinically as well as serologically. It concerns a child, which was born after 35 weeks of gestation. Two days after the delivery the mother showed typically efflorescences of Herpes simplex infection in the abdominal region. On the fifth day after birth the child showed a vesico-bullous exanthema beginning on the head and spreading out on breast and back. On day 14th a serious sepsis like pattern of the disease with respiratory insufficiency and encephalitic symptoms could be seen. Treatment with Vidarabinphosphat and Acyclovir-Natrium was without definite success. At the age of five months the child showed a pseudobulbar-paralysis with tetra-spasticity. The cranial computer-tomography demonstrated a distinct hydrocephalus e vacuo and the electroencephalography registered only sporadic brain activity. PMID- 2985862 TI - Seroepidemiology of HTLV-III (LAV) in the Federal Republic of Germany. AB - In 1984 10,281 sera were collected in the FRG and examined for antibodies to HTLV III (LAV) with an enzyme-linked immunosorbent assay and confirmative tests. Of the German AIDS patients 81% have antibodies. Individuals belonging to AIDS risk groups, homosexuals, haemophiliacs and i.v. drug abusers, have antibody frequencies between 25%-72%. The detection of HTLV-III antibodies in blood donors indicates that the virus is being transmitted by blood transfusions. PMID- 2985864 TI - Creatine kinase isoenzymes: biochemical findings after therapeutical embolization of the hepatic artery in two patients with liver metastasis of an islet cell carcinoma or a carcinoid tumor. AB - An atypical creatine kinase (CK) isoenzyme migrating cathodically to CK-MM in the electrophoresis was observed as response to the embolization of the right hepatic artery in two patients with liver metastasis. The time course of the atypical CK release was compared to that of tumor secretion products, i.e., insulin and 5 hydroxy-in-dol-acetic acid. The CK isoenzyme probably reflects a mitochondrial breakdown of the metastasis in question. The influence of the treatment on liver function was characterized by a marginal augmentation of ASAT and ALAT and by a small but significant leakage of mitochondrial glutamate dehydrogenase. In addition to the atypical CK, CK-BB, and CK-MB isoenzymes were found in the serum of the patient with primary carcinoid carcinoma. PMID- 2985865 TI - [Prevention of cholelithiasis]. PMID- 2985863 TI - Human cytomegalovirus infection: recent developments in diagnosis and epidemiology. AB - Cytomegalic inclusion disease (CID) is caused by a horizontally or vertically transmitted human herpes virus infection and may persist for life without obvious clinical symptoms. A serious course of horizontal primary and recurrent infections, however, is often observed in immunocompromised persons such as recipients of organ transplants and patients receiving fresh blood transfusions. Vertical infection may cause fetopathies. The human cytomegalovirus (HCMV) is thought to inherit an oncogenic potential as lately discussed for AIDS and M. Kaposi. Laboratory diagnosis of HCMV infection is performed by light microscopy (inclusion bodies), electron microscopy, virus isolation in cell culture, demonstration of viral DNA and antigen in clinical specimens, by histochemical methods (e.g. immunoperoxidase technique) and by DNA and peptide analysis for identification of different isolates and viral finger prints. Evaluation of cell mediated immunity in HCMV infection is performed quantitatively (assessment of Thelper/Tsuppressor ratios) or qualitatively (specific lymphocyte stimulation by the antigen). In most cases laboratory diagnosis is achieved by serological methods, i.e. demonstration and quantitation of HCMV-specific antibodies. In this context, a number of liquid- and solid-phase immunoassays have been developed, of which immunofluorescence and ELISA are most commonly used, besides complement fixation and passive haemagglutination. These procedures on the one hand allow the use of different antigen preparations as early and late viral proteins, and on the other hand permit a specific determination of different Ig classes and subclasses. A variety of assays has been established especially for determination of virus-specific IgM antibodies, which are predominantly found in active infection. These, however, at least in part may show non-specific results caused by interference of rheumatoid factor or IgG competition. Such problems have now been dealt with and are avoided by IgG precipitation or IgM immunosorption ("mu capture" technique). These recent methods allow an exact epidemiological identification of risk groups for CMV infection. Results from our laboratory revealed 13% HCMV-IgM positive patients among pregnant women, 16% IgM positive patients among renal transplant recipients, 4% IgM positive cases in patients after cardiosurgery and 1.7% IgM positives among prostitutes. The prevalence of HCMV infection as indicated by specific IgG antibodies was 56%, 90%, 83%, and 90%, respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2985866 TI - [Budd-Chiari syndrome in a patient with liver cirrhosis and cancer]. PMID- 2985867 TI - [Surgery of carotid gland tumors]. PMID- 2985868 TI - [Adrenal cortical function in chronic glomerulonephritis]. PMID- 2985869 TI - Surface contact inhibits neutrophil superoxide generation induced by soluble stimuli. AB - Human neutrophils in suspension undergo a metabolic burst and generate reactive O2- metabolites upon exposure to many soluble and particulate stimuli. They can also be stimulated to produce O2- when in contact with surfaces. We found that when neutrophils were allowed to settle into protein-coated surfaces the amount of O2- they generated varied with the nature of the protein: IgG greater than bovine serum albumin greater than plastic greater than gelatin greater than serum greater than collagen. However, when polymorphonuclear leukocytes were permitted to settle onto a surface and then were stimulated with either phorbol myristate acetate or N-formyl-methionyl-leucyl-phenylalanine the O2- response was greatly diminished compared to control cells that were exposed to the stimulus in suspension. In contrast, superoxide production in response to the particulate stimulus opsonized zymosan was similar in both suspended and settled neutrophils. The degree of inhibition was not related to the degree of adherence since the diminished response occurred with all of the surfaces tested and in the presence of cytochalasin B. Onset of inhibition was very rapid as was recovery when cells were resuspended. Whereas production of O2- was greatly inhibited by surface contact, release of lysosomal enzymes was only slightly affected. The effect of surface contact did not appear to be mediated via activation of adenylate cyclase since the combination of a phosphodiesterase inhibitor and exogenous dibuteryl cyclic adenosine monophosphate did not inhibit phorbal myristate acetate O2- production, but surface contact did. These data indicate that surface contact such as would occur during diapedesis and chemotaxis profoundly alters neutrophil behavior by an unknown mechanism and imply that observations made on polymorphonuclear leukocytes in suspension cannot be generalized to polymorphonuclear leukocytes in tissue. PMID- 2985870 TI - Screening for HTLV-III antibodies--a government blunder? PMID- 2985871 TI - Hormonal sensitivity of adenylate cyclase from human endometrium: modulation by estradiol. AB - In human endometria, a membrane-bound adenylate cyclase is present, which is recovered in high yield in a low-speed particulate fraction. Neither the specific activity of the enzyme nor the response to modifiers that act through the regulatory subunit of the complex, are modified during the proliferative or secretory phase of the cycle. Surprisingly, we found that in vitro treatment of secretory endometrial membranes with 17 beta-estradiol stimulates 3- to 4-fold the activity of adenylate cyclase. However this response does not occur on proliferative membranes. The activation by estradiol is independent of the presence of guanylylimidodiphosphate and is additive to that of the nucleotide. Possibly, as the consequence of the phenomenon, the concentration of cyclic AMP is significantly higher in curretage samples obtained from patients during the secretory rather than in the proliferative phase of the cycle. To our knowledge this is the first evidence of a target-cell membrane-directed effect of sex steroids in humans. PMID- 2985872 TI - The effect of ACTH on the plasma concentrations of the "hypertensinogenic" steroids, 17 alpha-hydroxyprogesterone and 17 alpha,20 alpha-dihydroxy-4-pregnen 3-one in sheep. AB - The plasma concentrations of 17 alpha-hydroxyprogesterone (17 alpha OHP) and 17 a'20 alpha-dihydroxy-4-pregnen-3-one (17 alpha 20 alpha OHP) have been measured in sheep during 5 days of ACTH administration at 20 micrograms/kg/day a rate of infusion known to produce hypertension. Five days of ACTH administration produced a progressive increase in plasma 17OHP from 0.45 +/- 0.12 to 128.9 +/- 28.4 nmol/l and in 17 alpha 20 alpha OHP from 0.54 +/- 0.15 to 73.1 +/- 7.2 nmol/l. Calculation of the blood production rate of both steroids during ACTH treatment confirms that the rates of infusion of 17OHP (3.0 mumol/h) and 17 alpha 20 alpha OHP (1.5 mumol/h) used to produce hypertension, when infused together with the other major ovine adrenocortical steroids, produced plasma concentrations in the range as found following administration at a rate to increase blood pressure. PMID- 2985873 TI - Influence of E. coli endotoxin on ACTH induced adrenal cell steroidogenesis. AB - The effect of endotoxin (lipopolysaccharide from E. coli) on isolated adrenocortical cells was examined. Lipopolysaccharide decreased the ACTH-induced steroidogenesis. This effect was shown by all corticotropin concentrations studied, and the longer the incubation time, the higher the effect produced. The rate of decrease of ACTH-induced steroidogenesis was dependent on the concentration of lipopolysaccharide in the medium. Binding of [125I]ACTH to adrenocortical cells was modified by lipopolysaccharide; this modification was related to a decrease of the ACTH-induced steroidogenesis. This effect supports the hypothesis of a direct interaction between lipopolysaccharide and the cell membrane with a concomitant distortion of the cell surface affecting the ACTH receptor sites of their environment. [14C]Lipopolysaccharide binds to isolated adrenocortical cells. Binding specificity was investigated by competitive experiments in the presence of various types of endotoxins, polypeptide hormones and proteins. Unlabelled lipopolysaccharide from the same bacterial strain and isolated under identical conditions than the labelled lipopolysaccharide exerted the strongest inhibitory activity. Unlabelled lipopolysaccharide of various strains different from that originating the labelled lipopolysaccharide exerted the less displacement. It would imply a certain kind of specificity but the decrease in the binding of lipopolysaccharide produced by ACTH and glucagon suggests the existence of non-specific interactions between lipopolysaccharide and cell membrane. PMID- 2985874 TI - Systemic scleroderma and small cell carcinoma of the lung. AB - A 64-year-old man diagnosed to have systemic scleroderma for 1 year developed small-cell carcinoma of the lung. Six additional cases obtained from the literature describing small-cell carcinoma occurring in patients with scleroderma are reviewed. PMID- 2985875 TI - Bronchial adenoma: surgical experience with long-term follow-up (4-17 years). AB - Of 16 patients with bronchial adenoma who were operated on at Beilinson Medical Center from 1967 to 1980, only three presented the "triad" of cough, hemoptysis, and recurrent pulmonary infections. In two patients the tumor was diagnosed incidentally and in five patients histological evidence of adenoma was made during bronchoscopy. One patient died of myocardial infarction following reoperation for bleeding, and one patient was lost to follow-up. The remaining 14 patients were followed for 4 to 17 years without evidence of local recurrence or distant metastases. We conclude that the long-term prognosis of patients with bronchial adenoma is excellent, and limited surgical procedure should be the treatment of choice whenever possible. PMID- 2985876 TI - 14th annual UCLA symposium. April 6-April 25, 1985. Abstracts. PMID- 2985877 TI - 14th annual UCLA symposium. Abstracts: Papilloma viruses: molecular and clinical aspects. PMID- 2985878 TI - Glucocorticoid receptors of normal and leukemic cells: role of proliferation conditions. AB - A published whole-cell binding assay of 3H-dexamethasone (3H-DEX) was combined with cell sedimentation analysis to investigate various factors influencing specific binding of glucocorticoids by leukemic lymphoblasts. Studies with mouse L1210 and human HL-60 cell lines revealed that glucocorticoid (GC) receptors accumulate during G1 phase of the cell cycle. In human lymphoblastic leukemia, the per cell receptor number was highest in cells in S and G2 phase and lowest in small, noncycling cells. In normal human white blood cells, GC receptor content was maximal in large lymphocytes and monocytes while no receptors were present in small lymphocytes. However, the receptor density of large lymphocytes was still 3 4 fold lower than of leukemic lymphoblasts of similar size. In L1210 cells the number of GC receptors decreased considerably in stationary cultures or following inhibition of cell cycle progression by dibutyryl cyclic AMP (dbc-AMP). Receptor content was also reduced in cells growing as ascites tumors in hosts with terminal disease. Accordingly, the receptor content in leukemic blasts appeared highly dependent on cell cycle distribution and on the proliferative status of the tumor cells. These findings may in part explain the large interpatient variation and the conflicting views regarding GC receptor content and prognosis in acute lymphoblastic leukemia. PMID- 2985879 TI - Isoenzyme studies in human leukemia-lymphoma cell lines--1. Carboxylic esterase. AB - The isoenzyme patterns of carboxylic esterase (E.C. 3.1.1.1) were studied in 74 proven human leukemia-lymphoma and 12 normal B-lymphoblastoid cell lines. These cell lines have been extensively phenotyped using poly- and monoclonal antibodies. Esterase isoenzymes were separated by isoelectric focusing and visualized by histo-cytochemical techniques. No leukemia-specific or (except for monocytes) blood cell type-specific isoenzyme or isoenzyme pattern could be detected. The monocytic element in some cell lines was characterized by a strong isoenzyme band which could be selectively and completely inhibited by sodium fluoride. The enzyme phenotypes were stably expressed in all subcultures of a given cell line and did not appear to have any cell cycle dependency. The leukemia-lymphoma cell lines have been subclassified into four major groups according to immunological parameters: T-cell, B-cell, myelomonocytic and non-T, non-B-cell. On the basis of immunological data the T-cell lines were assigned to five stages of differentiation. The number and staining intensity of the isoenzymes increased with differentiation of the T-cells paralleling the expression of immunological markers. The B-cell leukemia-lymphoma cell lines were divided into pre B-, B-, Burkitt lymphoma, multiple myeloma and hairy cell leukemia cell lines. Substantial variability among the isoenzyme patterns was detected ranging from immature profiles of pre B-cell lines to complete isoenzyme repertoires of multiple myeloma cell lines. No significant difference was seen between the isoenzymes of mature B-cell lines and normal B-lymphoblastoid cell lines. The most prominent feature seen in myelomonocytic cell lines was the monocytic band indicating a monocytic origin and separating the 'monocytoid' from the 'pure myeloid' cell lines. Considerable heterogeneity in the isoenzyme patterns was observed in the non-T, non-B cell groups which comprised erythroleukemia cell lines and cell lines arrested at a very early stage of lymphoid differentiation. These latter cell lines together with some T- and B cell lines shared the common characteristics of positivity for cALLA, TdT and Ia antigens and an immature, incomplete isoenzyme profile. The results support the notions of maturation arrest and normal gene expression in leukemic cell populations. Furthermore, the importance of biochemical studies as part of the multiple marker analysis could be demonstrated. PMID- 2985880 TI - Catalogue of Epstein-Barr virus (EBV) receptors on human malignant and non malignant hematopoietic cell lines. AB - Epstein-Barr virus (EBV) can induce a broad spectrum of hematological diseases, especially in immune deficient patients. We assayed for receptor for EBV (EBVR) using fluoresceinated viral particles on 44 human hematopoietic cell lines derived from patients with T, B, and non-T, non-B acute lymphocytic leukemia (ALL), non-lymphoid leukemia, Burkitt lymphoma, myeloma and several unique lines we and others have recently developed. All 31 EBV nuclear-associated antigen (EBNA) negative cell lines were of neoplastic origin. Seven of 13 EBNA-positive cell lines were of normal cell origin. Four of 25 non-B (surface immunoglobulin negative) EBNA-negative neoplastic cell lines were EBVR-positive. Three of six EBNA-negative B-cell (surface immunoglobulin positive) lines were EBVR-positive. Nine of 13 EBNA-positive Burkitt and non-Burkitt cell lines strongly expressed EBVR. Four EBNA-positive Burkitt lymphoma cell lines exhibited EBVR only to a limited degree. Studies of the cell lines for EBVR, complement receptors (CR) and surface immunoglobulin (SIg) revealed that presence of SIg does not obligate the presence of EBVR. Functional EBVR accompanied SIg among EBNA-negative cell lines. SIg-negative cell lines can possess EBVR. Fourteen of 16 EBVR-positive lines were also positive for CR. The EBVR assay is a useful tool for assessing the potential role of EBV in the induction of hematopoietic disorders. PMID- 2985881 TI - A computerized system for the detection and analysis of spontaneously occurring synaptic potentials. AB - A computerized system is described which converts analog, high-gain intracellular records from alpha motoneurons to digital data which are displayed on a graphics terminal and stored on magnetic media. Spontaneously occurring synaptic potentials are automatically detected and a quantitative analysis of their waveform parameters is computer-generated. PMID- 2985882 TI - Detection of synchrony in the discharge of a population of neurons. I. Development of a synchronization index. AB - A test for synchronization among the spike trains of muscle afferents or motor units is described which utilizes averages of neurograms and rectified neurograms. Synchronization is quantified by the increase of a synchronization index Is above a theoretical value for asynchrony. The dependence of the Is on signal amplitude and certain experimental conditions and a method of estimating confidence limits for the test are presented. PMID- 2985883 TI - Detection of synchrony in the discharge of a population of neurons. II. Implementation and sensitivity of a synchronization index. AB - This report describes the use of a synchronization index (Is; Hamm et al., 1985a) and its sensitivity to various forms and degrees of synchrony between spike trains. The dependence of the Is on signal-to-noise ratio, the number of synchronized spike trains and their degree of synchrony is shown in analog and digital simulations. These simulations and a comparison with peristimulus time histograms under conditions of induced synchrony reveal that the Is is a sensitive measure of synchronization in a population of spike trains. PMID- 2985884 TI - The mechanism of action of leukotrienes A4, C4 and D4 on human lung parenchyma in vitro. PMID- 2985885 TI - Changes of DNA repair mechanisms during the aging of the rat. AB - Recent studies have shown a significant reduction in DNA repair capacity in aging rats. Therefore we were interested in investigating which repair mechanism is concerned in this reduction. We investigated: excision repair (ER); single-strand break repair (SSBR); double-strand break repair (DSBR); and gamma-endonuclease susceptibility (ES) by means of the following methods: [3H]thymidine ([3H]dThd) incorporation into DNA after damage by N-methyl-N-nitrosourea (MNU); nucleoid sedimentation after damage by methyl methanesulfonate (MMS); neutral elution techniques after damage by 4-nitroquinoline-1-oxide (NQO); and determination of ES sites by velocity sedimentation in an alkaline sucrose gradient after damage by gamma-irradiation. Studies were done with male Sprague-Dawley rats aged 9, 18 and 28 months using nine different organs. We were able to determine a distinct age dependency of excision repair, a slight reduction of single-strand break repair, an elevation of gamma-endonuclease susceptible sites and no significant change in double-strand break repair in the course of aging. Therefore we see a shift in the pattern of DNA repair: in old age strand break repair mechanisms become more important, while repair replication is reduced. From this we can conclude that genetic expression is altered during the aging process, with all the consequences for the disposition toward certain diseases. PMID- 2985886 TI - Impairment of glutamate uptake and absence of alterations in the energy transducing ability of old rat brain mitochondria. AB - The proton electrochemical gradient has been measured in old brain mitochondria isolated from 2- or 24-month-old rats with the use of different respiratory substrates. With succinate as substrate, neither the respiratory rate, membrane potential or delta pH varied with age, indicating that the dielectric strength of the mitochondrial membrane was unaltered in old animals. The ohmic behavior of the membrane was tested in experiments in which the respiratory rate was partially inhibited by malonate, and was found to be unchanged with age. When glutamate plus malate were used as substrates, the respiratory rate was substantially reduced, and a drastic decrease in glutamate uptake was observed in old rat brain mitochondria. PMID- 2985887 TI - [Hepatic angiosarcoma and cholangiocarcinoma in a patient with thorotrast deposits]. PMID- 2985888 TI - Acute viral encephalitis. AB - Clinical manifestations of acute viral encephalitis are described, differential diagnoses are outlined, and a diagnostic approach is recommended. Encephalitic syndromes caused by arboviruses, herpesviruses, enteroviruses, and parainfectious processes are discussed. PMID- 2985889 TI - [Focal epithelial hyperplasia (Heck disease)]. AB - A case of focal epithelial hyperplasia (Heck's disease) in a nine years old caucasian girl is described. The disease is characterized by multiple flat verruca like elevations of the oral mucosa. The histological findings were acanthosis and papillomatosis with elongated anastomosing rete ridges. We also observed vacuolated cells with a hypercromatic marginated nucleo. In this paper the etiology is also discussed and we agreed that focal epithelial hyperplasia is an HPV, induced disease. Local application of vitamin A acid for three months was unsuccessful. PMID- 2985890 TI - [Merkel-cell tumor]. AB - A Merkel cell tumor of the preauricular region in a seventy-seven years old woman is described. Histological, electronmicroscopic and tissular hormonal studies, were made. From the pathological point of view, a nodular and trabecular pattern of oval or elongated cells with poor basophilic cytoplasma is characteristic. Large nuclei with irregular, oval prominent nucleoli are also present. Electron microscopy shows oval and polygonal cells of irregular nuclei with one or multiple prominent nucleoli. In the cytoplasm of these cells, microfilaments and dense core granules with clear halo are found. The tissular hormonal determinations were normal. Two years after the surgical treatment, the patient remains asymptomatic. PMID- 2985891 TI - Erythrocyte cationic transport systems in normal male and female volunteers. AB - The erythrocyte concentrations and the transmembrane fluxes of sodium and potassium were investigated in normal men and also in twenty normal women during the two stages of the menstrual cycle. Half of them were taking the contraceptive pill and the other half were not. In women with a normal menstrual cycle, the erythrocyte sodium concentration (Naic) and the ouabain-insensitive total potassium efflux were lower in the luteal than in the follicular phase. Intracellular potassium concentration (Kic), ouabain-sensitive 86rubidium-uptake and the furosemide-sensitive Na+ and K+ efflux did not differ significantly between the two periods of the cycle. No cycle-related variation in Naic or Kic was observed in women using the contraceptive pill. In these women, however, the ouabain-sensitive 86rubidium-uptake was increased in the second part of the menstrual cycle. Compared to men, the intra-erythrocyte sodium concentration was lower in women during the second stage of the menstrual cycle. These two groups were similar for Na+, K+-ATPase pump activity estimated from the ouabain sensitive 86rubidium-uptake and for the furosemide-sensitive sodium and potassium efflux. Women in the first stage of the menstrual cycle had intra-erythrocyte sodium concentration similar to men, but their furosemide-sensitive sodium efflux was lower. No significant difference was observed in the intraerythrocyte potassium concentration and transmembrane fluxes of potassium in men and women in either stage of the menstrual cycle. We therefore conclude from this study that one should take into account sex-related variability when studying cationic fluxes and concentrations in red blood cells of men and women. PMID- 2985892 TI - Effect of exogen pentagastrin on mastocyte degranulation, gastric cAMP and intragastric pH in Shay rats. AB - The degranulation of mastocytes, gastric cAMP and intragastric pH after the administration of 125 micrograms/kg exogen pentagastrin in Shay rats was studied. The results showed a decrease in the number of mastocytes in the gastric mucous membrane in the first hour analysed with a highly significant increase afterwards. There was a significant increase in the values of cAMP which corresponded to an acidification of intragastric pH. From these results it may be deduced that the increase in the release of histamine, as reflected by the decrease in the number of mastocytes, provokes the increase in the gastric levels of cAMP, which would in turn be responsible for the increase in the acid secretion provoked by the administration of exogen pentagastrin. PMID- 2985893 TI - Mechanism of presynaptic inhibition of cholinergic transmission in guinea-pig ileum by adenine nucleotides. AB - This study was designed to determine the mechanism of action of 2'-, 3'- and 5' substituted adenine nucleotides on cholinergic transmission in guinea-pig ileum. Segments of ileum were continuously stimulated at 0.2 Hz and IC50 values of nucleotides for inhibition of twitch responses determined. All the nucleotides studied inhibited transmission, the effect being antagonised by 30 X 10(-6) M theophylline. The inhibitors of nucleoside transport, HNBTGR and dipyridamole, potentiated responses to all the nucleotides. Addition of adenosine deaminase reduced responses only to adenosine and 5'-AMP, while inhibition of adenosine deaminase with deoxycoformycin potentiated responses only to 5'-AMP and 5'-ADP. It was concluded that all the nucleotides studied inhibit cholinergic transmission through an action at P1-purinoceptors. However, it was not possible from this study to conclude whether these actions were direct or indirect following their hydrolysis to adenosine. PMID- 2985894 TI - [Recurrent benign tumors of the parotid gland and their tendency for becoming malignant]. AB - In 257 parotid adenomas lumpectomy led to tumour recurrence in 49.2%, superficial parotidectomy in 5.7% and total parotidectomy in 5.3%. 40 patients with recurrent tumours experienced eventually eight true malignant neoplasms. There were five carcinomas ex pleomorphic adenomas, one true malignant mixed tumour, and two undifferentiated carcinomas. Malignant transformation usually occurred with the 4th recurrence after 15.8 years. To prevent recurrence and secondary malignancy in parotid adenomas the lumpectomy technique should be omitted. PMID- 2985895 TI - [Intracranial and cervical elimination of carotid circulation as a prerequisite for operability of extensive neck tumors]. AB - Large malignant tumours of the neck with an infiltration of the joint or internal carotid walls are mostly regarded as inoperable, if not as incurable. The cooperation between a neurosurgeon and an ENT surgeon opens the possibility, by elimination of the carotid flow, to effect a radical removal even of exceptionally extended tumours, thus offering a chance of survival for otherwise incurable patients. PMID- 2985896 TI - [Integrated and nonintegrated hepatitis B virus DNA in liver tissue]. AB - Liver tissue was taken in eight patients with virus hepatitis B and one patient with liver carcinoma by biopsy, as well as in seven other patients at post mortem. HBV-DNA was measured in these tissue specimens by hybridization. In four out of eight patients who had had biopsy, HBV-DNA could be found; in two patients it was present in integrated form. The same was true for the tumor tissue stemming from the patient with liver carcinoma. In five out of eight liver tissue specimens taken at post mortem HBV-DNA could be demonstrated as well; it was integrated into the host genom in two cases. It may be important to find out in patients with chronic hepatitis virus infection, if HBV-DNA is present in free or integrated form before antiviral treatment is considered. PMID- 2985897 TI - [Somatostatinoma of the liver]. AB - A case report is given of a 45 year old patient, who had a tumor of the liver, pain in the upper abdomen and relapsing fever. Surgery was done and part of the liver was removed. On microscopic examination a somatostatinoma was found. The clinical symptoms of this very rare type of tumor, which belongs to the APUD cell system, is discussed as well as therapeutical implications. PMID- 2985898 TI - Untreated borderline-leprosy in the ulnar nerve: light and electron microscopical studies. PMID- 2985899 TI - Testicular angiotensin I-converting enzyme (E.C. 3.4.15.1). AB - In the two mammalian species (i.e., rabbit and rat) in which it has been studied to date, testicular angiotensin I-converting enzyme possesses distinct physicochemical and immunological properties, and a susceptibility to hormonal regulation that makes it a unique isozyme of the converting enzyme ordinarily distributed throughout the body. The testicular isozyme appears to be a lower molecular weight version of the pulmonary enzyme, with similar, although not identical, catalytic properties. The testicular isozyme is under androgenic control and is associated with germinal cells. Although its function has yet to be elaborated, the testicular isozyme provides an excellent model for the study of tissue-specific regulation of carboxypeptidases. PMID- 2985900 TI - Visualization by positron emission tomography of the apparent regional heterogeneity of central type benzodiazepine receptors in the brain of living baboons. AB - The feasibility of visualizing the heterogeneity of benzodiazepine (BDZ) receptors in the brain of living baboons was investigated using Positron Emission Tomography. Ethyl 8-fluoro-5,6-dihydro-5-methyl 6-oxo-4H-imidazo (1,5-a) (1, 4) benzodiazepine-3-carboxylate (RO 15 1788) labelled by carbon 11 (11C-RO 15 1788) was I.V. injected for the "in vivo" labelling of the central type BDZ receptors. Displacement experiments were performed 20 minutes after the administration of the radioligand by two different cold drugs: RO 15 1788 which has an equal affinity for central type BDZ receptors, and propyl B-Carboline-3-carboxylate (B CCP) which favours the sites located in the cerebellum. Different sensitivities to these two drugs displacement of 11C-RO 15 1788 binding "in vivo" were observed: on the one hand in the regional localization of the displacement, and on the other hand, in the amount of the radioactivity displaced. The apparent interregional heterogeneity of the displacement seen in the cerebellum and in the temporal cortex are discussed in terms of discrepancies observed "in vitro" at physiological temperature, between cerebellar and non-cerebellar BDZ central type binding sites. PMID- 2985901 TI - Suppression of basal and stress-induced prolactin release and stimulation of luteinizing hormone secretion by alpha-melanocyte-stimulating hormone. AB - alpha-MSH and beta-endorphin, both synthesized from a common precursor, have opposite behavioral actions. In order to determine if these peptides have opposite effects on pituitary function, basal LH secretion and basal and stress induced prolactin release were studied in adult male rats after intraventricular injection of alpha-MSH. Each rat also received intraventricular saline in order to serve as its own control. 18 micrograms alpha-MSH stimulated plasma LH from 16.5 +/- 2.5 (SEM) ng/ml to a peak of 27.2 +/- 4.0 and 26.0 +/- 4.9 ng/ml at 5 and 10 min, and suppressed prolactin from 3.5 +/- 0.7 ng/ml to 1.3 +/- 0.1 and 1.2 +/- 0.1 ng/ml at 15 and 30 min. Intraventricular alpha-MSH also significantly blunted the prolactin rise associated with the stress of swimming. 10 and 20 min after the onset of swimming, prolactin levels in rats pretreated with alpha-MSH were significantly diminished: 7.4 +/- 1.5 and 6.5 +/- 2.0 ng/ml vs 23.8 +/- 3.6 and 15.2 +/- 2.8 after normal saline. Similarly, des-acetyl alpha-MSH which is the predominant form of alpha-MSH in the hypothalamus, diminished the stress induced prolactin rise from 18.4 +/- 5.3 and 11.2 +/- 3.4 ng/ml at 10 and 20 min to 10.0 +/- 2.4 and 5.5 +/- 1.6 ng/ml. We conclude that centrally administered alpha-MSH stimulates LH and suppresses basal and stress-induced prolactin release in male rats. These actions are opposite to those previously shown for beta endorphin and suggest that alpha-MSH may antagonize the effects of beta-endorphin on pituitary function. PMID- 2985902 TI - Differentiating opioids by their pupillary effects in the rat. AB - Pupillary effects of several opioids were examined as part of a broader in vivo study of multiple opioid receptors in the rat. Agonist activity, stereospecificity, and naloxone sensitivity were determined by methadone (Me), ethylketocyclazocine (EK), and N-allylnormetazocine (SKF 10,047), selected for their purportedly predominant actions at mu, kappa, and sigma receptors, respectively. After an acute, subcutaneous injection of each drug, pupil area and fluctuations in pupil size were measured by means of an infrared video pupillometer on line with a microcomputer data processing and storage system. Despite differences in the magnitude of the response, each opioid tested produced an increase in pupil size which was stereospecific, independent of behavioral responses to the drugs and, for 1-Me and 1-SKF 10,047, dose-related. Other differences among the opioids were found in their ability to induce fluctuations (1-Me and 1-EK) and a pendular nystagmus (1-SKF 10,047 only), and in their sensitivity to naloxone. Although 1.0 mg/kg naloxone completely reversed 1-Me induced mydriasis, 10 mg/kg was needed to reverse 1-EK, and this dose only partially antagonized 1-SKF 10,047. These characteristic patterns of pupillary responses to opioids in terms of agonist activities and naloxone sensitivities indicate that the different opioid receptor types subserve different functions with respect to pupillary control. PMID- 2985903 TI - Binding and processing of epidermal growth factor in Panc-I human pancreatic carcinoma cells. AB - The binding of 125I-labeled epidermal growth factor (EGF) was studied in Panc-I human pancreatic carcinoma cells. At 37 degrees C, binding was rapid and associated with marked endocytosis of the ligand. Bound EGF was sequentially converted to a number of more acidic species as follows: pI 4.55 to pI 4.2, to pI 4.35, to pI 4.0. EGF internalization and processing were blocked at 4 degrees C. EGF did not alter cell growth when Panc-I cells were incubated in the presence of 2 to 10% serum. In contrast, when the serum concentration was lowered to 0.1%, EGF significantly enhanced cell replication after 6 days of culture. PMID- 2985904 TI - Effects of dimaprit on growth and differentiation of human promyelocytic cell line, HL-60. AB - The human promyelocytic cell line HL-60, differentiates in response to a variety of agents including dibutyryl cAMP and agents which increase intracellular cAMP concentrations (phosphodiesterase inhibitors, PGE2, and cholera toxin). HL-60 is also known to be rich in H2 -histamine sensitive adenylate cyclase activity. The present study was therefore designed to test the effects of H2-stimulation on growth and differentiation of HL-60 using the potent H2 agonist dimaprit. Dimaprit markedly increased cAMP production in a dose-dependent manner reaching maximal levels after 30-60 minutes. Intracellular cAMP levels decreased thereafter and by 24 hours were approximately 2-3 fold increased above control. Intracellular cAMP levels were not altered by dimaprit (10(-7)M to 10(-4)M) at 4 days in culture compared to either untreated HL-60 cells or dimethylsulfoxide (DMSO) (1.3%) treated cells. While exponential growth was unaltered by dimaprit (10(-7)M to 10(-4)M) as compared to control, dimaprit induced i) morphologic maturation to the myelocyte and metamyelocyte form with no differentiation seen beyond the metamyelocyte even after 6 days in culture, ii) increased NBT reductase activity and iii) dose-dependent increase in lysozyme activity which could be completely blocked by cimetidine, a specific H2 antagonist. Dimaprit induced differentiation of HL-60 cells was associated with an initial but transient increase in intracellular cAMP production. Maturation beyond the metamyelocyte stage was not observed. Acquisition of NBT reductase and lysozyme activity correlated with morphologic maturation. PMID- 2985905 TI - [Gamma scintigraphy of lymphatic nodes with metastatic involvement]. AB - The authors indicate that according to the degree of informative value all the methods used in this study can be listed as follows: the method of combined use of positive and negative scintigraphy, the method of positive scintigraphy with Ga67-citrate (the lymph nodes above the diaphragm) and Tc99-pertechnetate (the lymph nodes below the diaphragm), the method of indirect lymphography with colloids. The main indices of the radionuclide methods in the diagnosis of the lymph node metastatic involvement are presented. PMID- 2985907 TI - [Pulmonary thromboembolic diseases in Ivory Coast (apropos of 13 cases)]. AB - From January 1983 to March 1984, 13 cases of pulmonary thrombo-embolic disease (PTE) were reported at the Abidjan cardiology Institute. Disease frequency is higher than usually reported in Africa. The reason is that angiography was carried out as soon as the clinical diagnosis was suspected. Radiologic examination allowed to locate with accuracy the level of the thrombosis. Under heparin, a favourable course of the disease was noted any time a phlebitis of lower extremity was involved. Two deaths occurred among the 13 cases. Few differences are noted in Africa and temperate zones as regard to disease behaviour. However, frequency of obstetrical and infectious aetiology is to be outlined. PMID- 2985906 TI - [Carcinoembryonic antigen and hormonal polypeptides in patients with lung cancer]. AB - The paper is concerned with the evaluation of the diagnostic value of a tumor marker complex determined by in vitro analysis at primary diagnosis of lung cancer, during early postoperative follow-up and subsequent monitoring for early detection of disease progression. PMID- 2985908 TI - [Acute disseminated lupus erythematosus in Ivory Coast (apropos of 9 cases)]. AB - Nine cases of systemic lupus erythematosus (S.L.E.) among ivoirian Africans seen over a period of eleven years were studied. The pattern of clinical presentations is similar of that seen elsewhere. Photo-sensitivity is not described as in american Negroes. Death is usually due to severe infection. The incidence of the disease among populations of different african countries are compared. The low prevalence 0,18 p. mille may be a function of failure to diagnose the more mild cases. PMID- 2985909 TI - [Microbiological study performed on 1,998 vaginal swabs]. AB - From 1981 to 1983, 1998 vaginal samples have been microbiologically examined in Abidjan. The average age was 23. Pathogen agents take often the place of normal lactobacillus flora. The presence of most of the microbian germs (Enterobacteriaceae, Neisseriae, Trichomonas, Corynehactery) was not increased by the pregnant state. But yeast and Candida albicans in particular, were found significantly more frequent in pregnant women. Candida albicans, serotype B, was, by far, the most frequently discovered. It has offered a real resistance against 5 fluorocytosin, as it can be observed in Europe. In vitro, ketoconazole appeared less efficient than econazole. On the contrary, nystatin and amphotericin B presented a great and constant activity against all yeasts of Candida genus. PMID- 2985910 TI - [Appearance of human anthrax in Ivory Coast forests]. AB - Eight cases of human anthrax were reported in 1983 from Ivory Coast Republic, all of them in the tropical forest belt (7 degrees 30' N). Clinical forms were: cephalic cutaneous (4 cases), intestinal (1 case) and very likely neurological (3 cases). Human disease was correlated to epizootic anthrax occuring in sheeps and goats within this area. Bacteriological cultures of animal material confirmed the diagnosis. This report of human anthrax is the second one coming from Ivory Coast Republic but the first one from the western Africa forest belt. PMID- 2985911 TI - Guar by-product improves carbohydrate tolerance in rats. AB - These studies were designed to assess the effect of guar by-product (GBP) upon carbohydrate tolerance in rats. Both 1% and 10% GBP suspensions administered immediately before a glucose challenge (1 g/kg body weight) caused a 31% reduction in the integrated plasma glucose response area during a 180 min test. Because 10% GBP caused a flattened glucose response curve, it was studied further. Ten percent GBP caused a similar reduction in plasma glucose during a 360-minute test; similar plasma insulin levels were measured in both 0% and 10% GBP groups throughout the 360-minute test. Ten percent oat bran and 10% soy bran had no effect upon plasma glucose responses. Ten percent GBP was effective in lowering glycemic excursions after sucrose and starch challenges. Rats fed pellet fractions from water and acid treated GBP had the lowest plasma glucose excursions after a glucose challenge; simulated gastric juice treatment of GBP reduced the hypoglycemic activity and simulated pancreatic juice treatment destroyed it. These studies demonstrate the ability of GBP suspensions to significantly reduce plasma glucose excursions after carbohydrate challenge. PMID- 2985912 TI - Binding assays for epidermal growth factor. PMID- 2985913 TI - Radioligand assay for angiotensin II receptors. PMID- 2985914 TI - Wheat germ lectin-sepharose affinity adsorption assay for the soluble glucagon receptor. PMID- 2985915 TI - Covalent labeling of the hepatic glucagon receptor. PMID- 2985916 TI - Assaying binding of nerve growth factor to cell surface receptors. PMID- 2985917 TI - Peptide hormone receptors in intracellular structures from rat liver. PMID- 2985919 TI - Assay for the glucagon receptor. PMID- 2985918 TI - Isolation and functional aspects of free luteal cells. PMID- 2985920 TI - Culture and characteristics of hormone-responsive neuroblastoma X glioma hybrid cells. PMID- 2985921 TI - Primary glial cultures as a model for studying hormone action. PMID- 2985922 TI - Study of receptor function by membrane fusion: the glucagon receptor in liver membranes fused to a foreign adenylate cyclase. PMID- 2985923 TI - Isolation of ACTH-resistant Y1 adrenal tumor cells. PMID- 2985924 TI - Induction of glucagon responsiveness in transformed MDCK cells unresponsive to glucagon. PMID- 2985925 TI - Characterization of hormone-sensitive Madin-Darby canine kidney cells. PMID- 2985926 TI - Construction of hormonally responsive intact cell hybrids by cell fusion: transfer of beta-adrenergic receptor and nucleotide regulatory protein(s) in normal and desensitized cells. PMID- 2985927 TI - Assays for calcitonin receptors. PMID- 2985928 TI - Analysis of hormone-induced changes of phosphoinositide metabolism in rat liver. PMID- 2985929 TI - Assay for parathyroid hormone receptors. PMID- 2985930 TI - Effect of bradykinin on prostaglandin production by human skin fibroblasts in culture. PMID- 2985931 TI - Direct chemical measurement of receptor-mediated changes in phosphatidylinositol levels in isolated rat liver plasma membranes. PMID- 2985932 TI - Gastrin receptor assay. PMID- 2985933 TI - Assay for the cholecystokinin receptor on pancreatic acini. PMID- 2985934 TI - Assay of antibodies directed against cell surface receptors. PMID- 2985935 TI - Characterization of antisera to prolactin receptors. PMID- 2985937 TI - A monoclonal antibody to growth hormone receptors. PMID- 2985936 TI - Methods for studying the platelet-derived growth factor receptor. PMID- 2985938 TI - 5'-p-Fluorosulfonylbenzoyl adenosine as a probe of ATP-binding sites in hormone receptor-associated kinases. PMID- 2985939 TI - Demonstration of T antigens on the surface of cells transformed with primate polyoma viruses. AB - Primate polyoma virus-transformed hamster, mouse, and rat cell lines were examined by indirect immunofluorescence staining for cell surface-associated T antigens, by using a rabbit antiserum prepared against sodium dodecyl sulfate denatured large T antigen of simian virus 40 (anti-SV40-SDS-T serum). Positive surface staining was shown not only on SV40-transformed cells, but also on BK and JC virus-transformed cells. In contrast, normal cells and cells transformed with mouse polyoma-, human adeno-, and murine sarcoma viruses were negative. The data on SV40-transformed cells confirmed the reports of others demonstrating the cell surface location of SV40 large T antigen, and the data on BK and JC virus transformed cells proved that these cells have cell-surface T antigens that cross react with anti-SV40-SDS-T serum. PMID- 2985940 TI - A focus assay method for Japanese encephalitis virus using complement and anti virus serum. AB - A sensitive, quantitative, short-time, and reproducible focus assay for Japanese encephalitis (JE) virus is described. After 2 or 3 days of incubation, the infected cells were treated with anti-JE virus serum and complement, and subsequently stained with trypan blue; then clear foci were produced. This method made it easy to titrate the infectivities not only of all seven JE virus strains tested but also of West Nile (WN), Murray Valley encephalitis (MVE), and St. Louis encephalitis (SLE) viruses using hyperimmune anti-JE virus serum for the latter. Moreover, even cell lines which hardly formed plaques by the agar overlay method easily produced foci within 2 or 3 days by this method. PMID- 2985941 TI - Replication of group A coxsackieviruses in monkey kidney cells. PMID- 2985942 TI - Iodothyronines and iodotyrosines as hypothetical receptors for catecholamines and opiates. AB - The signs and symptoms of hyperthyroidism are similar to those of phaeochromocytoma and adrenergic stimulation. These changes have been attributed to increased beta receptor numbers in hyperthyroidism, resulting in adrenergic supersensitivity. Beta blockers are used in treatment of hyperthyroidism and inoperable phaeochromocytomas. Beta receptor numbers increase in direct proportion to the dose of thyroid hormone given. Hypothyroidism decreases the number of alpha and beta receptors with greater reduction in beta receptor numbers. Beta responses predominate in hyperthyroidism and alpha responses in hypothyroidism. Hypothyroid patients show increased sensitivity to opiates while hyperthyroid patients are tolerant to opiates. In the present paper it is proposed that the various iodothyronine amino acids are incorporated into the various catecholamine and opiate receptors, providing the phenyl binding sites in the receptors. T3 and T4 are proposed as part of beta 1 chronotropic and inotropic receptors and those mediating vasodilation. T2 and rT3 are proposed as part of alpha 1 and alpha 2 receptors and dopamine and opiate receptors. T2 is probably incorporated into beta 2 receptors causing bronchodilation. The different structural features of the various amino acids would account in part for the selectivity of various drugs for various receptors. PMID- 2985945 TI - How to market yourself. PMID- 2985943 TI - Calcium, parathyroid hormone and phospholipid turnover of human red blood cells. AB - PTH enhances entry of calcium into RBC, stimulates their Ca ATPase and increases osmotic fragility. The effect on the latter occurs only in the presence of calcium. The mechanisms of action of PTH and those mediating their increased calcium influx into RBC are not evident. It is possible that calcium, PTH or both affects phospholipid turnover of RBC and results in production of ionophoric compounds such as phosphatidic acid which in turn allows calcium movement. The present study examined the effect of calcium (1 mM), 1-84 PTH (50 U/ml) and calcium and PTH on phospholipid contents and 32P incorporation into phosphoinositides of human RBC and RBC ghosts. Calcium produced significant decrements in 32P incorporation into phosphatidylinositol, di- and triphosphatidylinositol and an increase into phosphatidic acid. PTH decreased the 32P into phosphatidylinositol only. Both PTH or calcium caused a significant increase in RBC content of phosphatidylserine. The data show that calcium alone alters phospholipid turnover of RBC and such an effect may mediate the PTH induced calcium influx. Alternatively, PTH may enhance entry of calcium into RBC independent of the effect of calcium on phospholipid turnover and the increase in cytosolic calcium would then alter phospholipid turnover which in turn would further facilitate the effect of PTH on calcium influx. The increase of phosphatidylserine by PTH may increase rigidity of RBC membrane and enhance osmotic fragility. PMID- 2985944 TI - Effects of (Sp)- and (Rp)-adenosine cyclic 3',5'-phosphorothioates on electrolyte excretion by the isolated perfused rat kidney. AB - We studied the effects of the (Sp) and (Rp) diastereomers of the phosphorothioate analogue of cyclic AMP (cAMPS) on the excretion of electrolytes in the isolated perfused rat kidney. cAMPS is highly permeant across the peritubular cell membrane and is not metabolized by the rat kidney (Coulson et al., Life Sci. 32: 1489-1498, 1983). Addition of 10 microM cAMPS(Sp) to the perfusate resulted in a significant phosphaturia, bicarbonaturia, magnesuria and natriuresis and no change in renal vascular resistance or glomerular filtration rate. Fractional excretion of calcium was elevated by cAMPS(Sp) but proportionately less than the fractional excretion of sodium so that the ratio of calcium to sodium clearances was significantly lowered. cAMPS(Rp) 10 or 100 microM was without effect on renal electrolyte excretion. The parathyroid hormone-like effects of the (Sp) diastereomer are consistent with its known ability to activate protein kinase. PMID- 2985946 TI - Unusual neurocutaneous syndromes. AB - During the last several decades, the number of these rare heterogeneous disorders described as neurocutaneous syndromes has significantly increased. The criteria for inclusion have become more general, although most of the disorders have dysplastic features, many do not have a tendency for tumor formation. Further, not all of these disorders are heritable and some are reported in only one or two families. Nonetheless, there is some merit for the clinician, geneticist, and embryologist to consider these anomalies of embryologic development on some common ground. Knowledge of these syndromes is important to correctly establish the diagnosis and prognosis. In cases of the heritable disorders, genetic counseling is essential. Finally, these disorders are of theoretic importance to all biologists, as the exchange of information will be essential to ultimately unravel the reasons for their cause. PMID- 2985947 TI - Myoclonus. AB - Myoclonus in childhood may be a primary condition, or it may be associated with a number of identifiable conditions. Successful therapy is dependent on diagnosis of underlying conditions and subsequent choice of appropriate drugs. PMID- 2985948 TI - Covalent labeling of the beta-adrenergic ligand-binding site with para (bromoacetamidyl)benzylcarazolol. A highly potent beta-adrenergic affinity label. AB - para-(Bromoacetamidyl)benzylcarazolol (pBABC) was synthesized and found to be an extremely potent affinity label for beta-adrenergic receptors. Its interaction with mammalian (rabbit and hamster lung) and nonmammalian (turkey and frog erythrocyte) beta-adrenergic receptors was similar, displaying EC50 values of 400 900 pM for inhibiting 125I-cyanopindolol binding to these receptors. pBABC reduced the number of beta-adrenergic receptors in frog erythrocyte membranes, without any change in the affinity of the remaining sites for [125I]iodocyanopindolol. pBABC has been radioiodinated. As assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, this affinity probe specifically labeled the beta-adrenergic peptide of a purified preparation of hamster lung, with high efficiency (approximately 40%) and with a pharmacological specificity characteristic of an interaction at the beta 2-adrenergic receptor ligand-binding site. Comparison of the proteolyzed products derived from purified receptor labeled with [125I]pBABC and with the photoaffinity agent [125I]p azidobenzylcarazolol suggested that covalent labeling of the beta-adrenergic receptor by these probes occurs at similar domains of the beta-adrenergic receptor. Because of the much higher level of incorporation of this affinity probe as opposed to photosensitive probes, pBABC should prove to be a useful tool for structural studies of purified beta-adrenergic receptors. PMID- 2985949 TI - Effects of tunicamycin on the expression of beta-adrenergic receptors in human astrocytoma cells during growth and recovery from agonist-induced down regulation. AB - Tunicamycin, which inhibits formation of asparagine-linked glycoproteins, caused a concentration-dependent blockade of beta-adrenergic receptor (beta-AR) accumulation in 1321N1 human astrocytoma cells during growth in culture. A concentration of tunicamycin (0.1 microgram/ml) that inhibited receptor accumulation and [3H]mannose or [3H]glucosamine incorporation into glycoproteins by 90% had only a small effect (10%) on [3H]leucine incorporation into protein, and reduced the rate of cell growth. Incubation in drug-free medium subsequent to treatment of 1321N1 cells with tunicamycin for 48 hr resulted in recovery of beta AR to control levels within an additional 48 hr. Exposure of cultures to isoproterenol (0.1 microM, 12 hr) caused an 80-90% loss of beta-AR in both pre- and postconfluent cultures; beta-AR recovered to control levels upon removal of isoproterenol. Although both tunicamycin and the protein synthesis inhibitor cycloheximide blocked beta-AR accumulation during growth of 1321N1 cells, neither agent inhibited the appearance of beta-AR during recovery from the down-regulated state in preconfluent cultures. However, cycloheximide, but not tunicamycin, blocked recovery of beta-AR after isoproterenol-induced loss of receptors in postconfluent cultures. In a previous report (Mol. Pharmacol. 26:424-429, 1984), we provided direct evidence that recovery of beta-AR from down-regulation in postconfluent cultures requires de novo synthesis of receptor protein. Thus, the results with tunicamycin are consistent with the idea that recovery of beta-AR in postconfluent cultures requires the synthesis of new beta-AR molecules, but as aglycoproteins that exhibit radioligand-binding characteristics similar to those of native glycoprotein beta-AR. PMID- 2985950 TI - Nonlinear relationship between alpha 1-adrenergic receptor occupancy and norepinephrine-stimulated calcium flux in cultured vascular smooth muscle cells. AB - To determine the relationship between vascular alpha 1-adrenergic receptor occupancy and receptor-coupled calcium flux, we have studied [3H]prazosin binding and l-norepinephrine-induced 45Ca efflux in cultured vascular smooth muscle cells isolated from the rabbit aorta. In a crude cellular homogenate, [3H]prazosin bound to a single high affinity site (Kd = 0.096 nM; Bmax = 105 +/- 15 fmol/mg of protein), whereas l-norepinephrine (NE) binding was best described by a two-site model with 43 +/- 8% of sites of high affinity (KH = 92 +/- 3 nM) and 57 +/- 7% of sites of low affinity (KL = 7460 +/- 4330 nM). NE-stimulated 45Ca efflux was concentration-dependent (EC50 = 108 nM) and potently inhibited by prazosin (IC50 = 0.15 nM), but not yohimbine (no inhibition at 10 microM). For the total receptor pool identified by [3H]prazosin binding, the relationship between receptor occupancy by NE and NE-stimulated 45Ca efflux was markedly nonlinear, such that 50% of maximum NE-stimulated efflux occurred with occupancy of only approximately 7% of receptors. Likewise, following irreversible inactivation of 69 +/- 5% of receptors by phenoxybenzamine, maximal NE-stimulated 45Ca efflux was decreased by only 8 +/- 2%. These two experimental approaches provide direct evidence for the presence in cultured rabbit aortic smooth muscle cells of a sizable pool of alpha 1-adrenergic receptors in excess of those needed for maximum NE-stimulated 45Ca efflux. This evidence of "spare" receptors, together with the finding of two affinity states of agonist binding, raises the possibility of functional heterogeneity of alpha 1-adrenergic receptors in this system. PMID- 2985951 TI - Catechol formation of fluoro- and bromo-substituted estradiols by hamster liver microsomes. Evidence for dehalogenation. AB - We have examined the validity of using fluorine-substituted estrogens as probes to assess the significance of 2- and 4-hydroxylation in estrogen-induced carcinogenesis in the hamster. Liver microsomes from castrated hamsters were incubated with 2-fluoro-, 4-fluoro-, or 2,4-difluoroestradiols and analogous bromo-substituted estradiols to determine the extent of 2- and 4-hydroxylation with these substrates. Estrogen 2- and 4-hydroxylase activity was determined by radioenzymatic assay, and the 3H-labeled monomethyl ether products were identified by high performance liquid chromatography. With unsubstituted 17 beta estradiol as substrate, 97% of the product formed was 2-hydroxylated, and 3% was 4-hydroxylated. The monosubstituted fluoroestradiols exhibited more than a 2-fold enhanced ability to form catechol estrogens compared with their corresponding bromoestradiols. Data presented herein indicate substantial defluorination when 2 fluoroestradiol was the substrate, which amounted to 36% of the total product formed, and 32% of the rate of 2-hydroxylation found with unsubstituted 17 beta estradiol as substrate. Interestingly, the rate of 4-hydroxylation was elevated 20- and 6.7-fold, respectively, when 2-fluoroestradiol and 2,4-difluoroestradiol were the substrates compared to the rate with 17 beta-estradiol. Moreover, both 4 fluoroestradiol and 2,4-difluoroestradiol exhibited at least a 1.6-fold greater rate of 2-hydroxylation compared with 17 beta-estradiol. In contrast, the rate of dehalogenation with 2-bromoestradiol was only 12% of that found with 2 fluoroestradiol. No debromination was obtained with 4-bromoestradiol, and essentially no catechols were formed using 2,4-dibromoestradiol as substrate with these hamster liver microsomes. These data clearly provide evidence for defluorination of these substituted estrogens, particularly at the C-2 position, and seriously hamper the use of fluoroestrogens in studies of hormonal carcinogenicity. PMID- 2985952 TI - Generation of helper-free amphotropic retroviruses that transduce a dominant acting, methotrexate-resistant dihydrofolate reductase gene. AB - We constructed several retroviruses which transduced a mutant dihydrofolate reductase gene that was resistant to methotrexate inhibition and functioned as a dominant selectable marker. The titer of dihydrofolate reductase-transducing virus produced by virus-producing cells could be increased to very high levels by selection of the cells in increasing concentrations of methotrexate. Helper virus free dihydrofolate reductase-transducing virus was also generated by using a broad-host-range amphotropic retroviral packaging system. Cell lines producing helper-free dihydrofolate reductase-transducing virus with a titer of 4 X 10(6) per ml were generated. These retroviral vectors should have general utility for high-efficiency transduction of genes in cultured cells and in animals. PMID- 2985953 TI - Functional analysis of the transcription control region located within the avian retroviral long terminal repeat. AB - We used several quantitative assays of in vivo transient gene expression to dissect the elements within the Rous sarcoma virus long terminal repeat (LTR) which constitute the retroviral transcription control region. Site-directed deletion mutagenesis was used to locate and define the enhancer and promoter elements within the LTR. In addition, we inserted exogenous DNA fragments into the LTR to examine the effects of position and sequence on the activity of these LTR transcriptional elements. The Rous sarcoma virus enhancer element, which we propose is located entirely within the LTR, was shown to activate both the beta globin and retroviral LTR promoters when located in cis. We observed a striking correlation between the degree of activation and the distance between the retroviral promoter and enhancer elements. The LTR promoter element mediated the activation effect of the enhancer element, as LTR deletion mutants containing only the enhancer and TATA box region expressed little activity. The promoter region encoded a low but significant level of transcriptional activity even in the absence of an enhancer. Overall LTR transcriptional activity declined sharply with increasing distance between the LTR promoter and initiator elements. These results shed light on both the importance of the spatial arrangement of the sequence elements within this eucaryotic transcription control region and on the functional interrelationship between these elements. PMID- 2985954 TI - Repeat arrays in cellular DNA related to the Epstein-Barr virus IR3 repeat. AB - We isolated clones and determined the sequence of portions of mouse and human cellular DNA which cross-hybridize strongly with the IR3 repetitive region of Epstein-Barr virus. The sequences were found to be tandem arrays of a simple sequence based on the triplet GGA, very similar to the IR3 repeat. The cellular repeats have distinct differences from the viral repeat region, however, and their sequences do not appear capable of being translated into a purely glycine plus-alanine protein domain like the portion of the Epstein-Barr nuclear antigen coded by IR3. Although the relationship between IR3 and the cellular repeats is left unclear, the cellular repeats have many interesting features. The tandem arrays are about 1 to several kilobases long, much shorter than satellite tandem repeats and larger than other interspersed, tandem repeats. Each of the repeats is a distinct variation, perhaps diverged from a common sequence, (GGA)n. This family is present in the genomes of all species tested and appears to be a ubiquitous feature of all higher eucaryotic genomes. PMID- 2985955 TI - gamma 2-Thymidine kinase chimeras are identically transcribed but regulated a gamma 2 genes in herpes simplex virus genomes and as beta genes in cell genomes. AB - True gamma or gamma 2 genes, unlike alpha, beta, and gamma 1 (beta gamma) genes of herpes simplex virus 1 (HSV-1), stringently require viral DNA synthesis for their expression. We report that gamma 2 genes resident in cells were induced in trans by infection with HSV-1 but that the induction did not require amplification of either the resident gene or the infecting viral genome. Specifically, to test the hypothesis that expression of these genes is amplification dependent, we constructed two sets of gamma 2-thymidine kinase (TK) chimeric genes. The first (pRB3038) consisted of the promoter-regulatory region and a portion of 5'-transcribed noncoding region of the domain of a gamma 2 gene identified by Hall et al. (J. Virol. 43:594-607) in the HSV-1(F) BamHI fragment D' to the 5'-transcribed noncoding and coding regions of the TK gene. The second (pRB3048) contained, in addition, an origin of HSV-1 DNA replication. Cells transfected with either the first or second construct and selected for the TK+ phenotype were then tested for TK induction after superinfection with HSV-1(F) delta 305, containing a deletion in the coding sequences of the TK gene, and viruses containing, in addition, a ts lesion in the alpha 4 regulatory protein (ts502 delta 305) or in the beta 8 major DNA-binding protein (tsHA1 delta 305). The results were as follows: induction by infection with TK- virus of chimeric TK genes with or without an origin of DNA replication was dependent on functional alpha 4 protein but not on viral DNA synthesis; the resident chimeric gene in cells selected for G418 (neomycin) resistance was regulated in the same fashion; the chimeric gene recombined into the viral DNA was regulated as a gamma 2 gene in that its expression in infected cells was dependent on viral DNA synthesis; the gamma 2-chimeric genes resident in the host and in viral genomes were transcribed from the donor BamHI fragment D' containing the promoter-regulatory domain of the gamma 2 gene. The significance of the differential regulation of gamma 2 genes in the environments of host and viral genomes by viral trans-acting factors is discussed. PMID- 2985956 TI - Rapid assay for extrachromosomal homologous recombination in monkey cells. AB - Most of the recombination assays based on the regeneration of selectable marker genes after transient infection or stable integration of DNA into mammalian cells are time consuming. We have used plasmids containing two truncated but overlapping segments of the neomycin resistance gene to rapidly quantitate and characterize the time course of extrachromosomal homologous recombination of DNA transfected into monkey COS cells. By transiently infecting cells with these recombination substrates, extracting Hirt DNA after 1 to 4 days, and transforming recombination-deficient Escherichia coli, we have shown that recombination between direct repeats occurs at frequencies of 1 to 4%. We have also used Southern blot analysis to directly characterize the recombination of this DNA in COS cells and to demonstrate that double-strand breaks in the region of homology increase recombination frequencies 10- to 50-fold. PMID- 2985957 TI - myc and src oncogenes have complementary effects on cell proliferation and expression of specific extracellular matrix components in definitive chondroblasts. AB - The effects of the avian viral oncogenes src and myc were compared for their ability to alter the differentiated phenotype and the proliferative capacity of definitive chondroblasts. As previously demonstrated, viruses carrying the src oncogene suppressed the synthesis of the chondroblast-specific products, type II collagen and cartilage-specific sulfated proteoglycan. In contrast, infection with MC29 and HB1 viruses, which carry the myc oncogene, did not suppress the synthesis of these normal differentiated cell products, but the infected cells exhibited an increased proliferative potential. The MH2 virus, which carries both the myc and mil oncogenes, both induced the suppression of these chondroblast specific products and increased cell proliferation. The implications of these results for cooperation between oncogenes and the multi-oncogene models for neoplastic transformation are discussed. PMID- 2985958 TI - Cloned mouse DNA fragments can replicate in a simian virus 40 T antigen-dependent system in vivo and in vitro. AB - Mouse liver DNA was cut out with BamHI and cloned into YIp5, which contained the URA3 gene of Saccharomyces cerevisiae in pBR322. Of the several plasmids isolated, two plasmids, pMU65 and pMU111, could transform S. cerevisiae from the URA- to the URA+ phenotype and could replicate autonomously within the transformant, indicating that mouse DNA fragments present in pMU65 or pMU111 contain autonomously replicating sequences (ARS) for replication in S. cerevisiae. Furthermore, to determine the correlation between ARS function in yeast cells and that in much higher organisms, we tried to challenge these plasmids with the simian virus 40 (SV40) DNA replication system. Of the two plasmids tested, the EcoRI-BglII region of pMU65 could be hybridized with a chemically synthesized 13-nucleotide fragment corresponding to the origin region of SV40 DNA. Both pMU65 (the EcoRI-BglII region cloned in pBR322) and its subclone pMU65EB could replicate semiconservatively, and initiation of DNA replication started from the EcoRI-BglII region when the replicating activity of these plasmids was tested in the in vitro SV40 DNA replication system we have established before. Furthermore, pMU65 and pMU65EB could replicate autonomously within monkey Cos cells which produce SV40 T antigen constitutively. These results show that a 2.5-kilobase fragment of the EcoRI-BglII region in pMU65 contains the ARS needed for replication in the SV40 DNA replication system. PMID- 2985959 TI - Characterization of a surrogate TATA box promoter that regulates in vitro transcription of the simian virus 40 major late gene. AB - The presence of a surrogate TATA box sequence located ca. 30 nucleotides upstream of the major late RNA start site at nucleotide position (np) 325 (Brady et al., Cell 31:625-633, 1982) has been confirmed, and its structural specificity has been determined by the generation of additional base substitution mutations at the KpnI restriction site (np 294) in cloned simian virus 40 DNA. Two mutants generated new RNA initiation sites upstream of the np 325 start site and continued to utilize the authentic start site, but with decreased efficiency. The replacement of either one or both cytosines by thymines at np 298 and np 299 specifically enhanced in vitro transcription from the np 325 start site by 430 and 800%, respectively. This enhancement was due to conversion of the simian virus 40 late promoter present in the wild-type virus to a sequence that is similar to the TATA box present in the simian virus 40 early promoter. PMID- 2985960 TI - Effects of alterations in cellular iron on biosynthesis of the transferrin receptor in K562 cells. AB - Treatment of K562 cells, a human erythroleukemia cell line, with desferrioxamine raised the levels of the receptor for transferrin (Tf) two- to threefold over that of the control cells. The levels of receptor were reduced by at least 50 and 35% of that of the control in cells treated with diferric Tf and ferric ammonium citrate, respectively. These changes were of total cellular receptors with no alteration in the proportion of receptors found on the cell surface. The half lives of the receptor were identical in cells treated with desferrioxamine, diferric Tf, or ferric ammonium citrate. Cells metabolically labeled with [35S]methionine showed a 2.5-fold increase in the rate of receptor synthesis when treated with desferrioxamine and a 35 and 65% decrease when treated with ferric ammonium citrate and diferric Tf, respectively. In vitro translations of polyadenylated mRNA isolated from cells incubated with desferrioxamine showed a 2.5-fold increase in translatable mRNA for the receptor, whereas treatment of cells with ferric ammonium citrate and diferric Tf resulted in a 25 and 50% reduction, respectively, in translatable mRNA for this receptor. PMID- 2985961 TI - Mapping of DNase I-hypersensitive sites in the 5' and 3' long terminal repeats of integrated moloney murine leukemia virus proviral DNA. AB - The chromatin state of integrated Moloney murine leukemia virus (M-MuLV) proviral DNA was investigated. Nuclei from M-MuLV-infected mouse NIH 3T3 cells were digested with limited amounts of DNase I, and hypersensitive (HS) sites were mapped by the indirect end labeling technique. Particular emphasis was placed on the 5' long terminal repeat (LTR), since viral transcription initiates there. M MuLV proviral DNA showed two strong DNase I-HS sites in the 5' LTR, one coincident with the transcription initiation (cap) site and the other with the transcriptional enhancers. Two weaker DNase I-HS sites were also detected in internal proviral DNA. The 3' LTR also showed a strong HS site in the region of the enhancers, but an HS site at the cap site of the 3' LTR was not detected. Thus, the chromatin configurations of the 5' and 3' LTRs of integrated M-MuLV proviruses appear to be different. The chromatin configuration of M-MuLV proviruses which contain LTR insertions of polyomavirus enhancer sequences was also studied. The 5' LTR of M-MuLV proviruses containing polyoma enhancer sequences substituted for the M-MuLV enhancers showed two strong HS sites, one in the polyoma sequences and one at the cap site. The 5' LTR of M-MuLV proviruses containing polyoma enhancer sequences inserted into the wild-type M-MuLV LTR between the cap site and the M-MuLV enhancers showed three HS sites. Two HS sites corresponded to those of the wild-type M-MuLV LTR, whereas the third mapped to the inserted polyoma sequences. The HS site associated with the inserted polyoma sequences was considerably stronger than the M-MuLV-associated HS sites. PMID- 2985962 TI - Similar 150-kilobase DNA sequences are amplified in independently derived methotrexate-resistant Chinese hamster cells. AB - We have isolated overlapping recombinant cosmids that represent 150 kilobases of contiguous DNA sequence from the amplified dihydrofolate reductase domain of a methotrexate-resistant Chinese hamster ovary cell line (CHOC 400). This sequence includes the 25-kilobase dihydrofolate reductase gene and an origin of DNA synthesis. Eight cosmids that span this domain have been utilized as radioactive hybridization probes to analyze the similarities among the dihydrofolate reductase amplicons in four independently derived methotrexate-resistant Chinese hamster cell lines. We have observed no significant differences among the four cell lines within the 150-kilobase DNA sequence that we have examined, except for polymorphisms that result from the amplification of one or the other of two possible alleles of the dihydrofolate reductase domain. We also show that the restriction patterns of the amplicons in these four resistant cell lines are virtually identical to that of the corresponding, unamplified sequence in drug susceptible parental cells. Furthermore, measurements of the relative copy numbers of fragments from widely separated regions of the amplicon suggest that all fragments in this 150-kilobase region may be amplified in unison. Our data show that in methotrexate-resistant Chinese hamster cells, the amplified unit is large relative to the dihydrofolate reductase gene itself. Furthermore, within the 150-kilobase amplified consensus sequence that we have examined, significant rearrangements do not seem to occur during the amplification process. PMID- 2985963 TI - Analysis of a transgenic mouse containing simian virus 40 and v-myc sequences. AB - Two plasmids, one containing the simian virus 40 (SV40) genome and the mouse metallothionein I gene and one containing the v-myc gene of avian myelocytomatosis virus MC29, were coinjected into mouse embryos. Of the 13 surviving mice, one, designated M13, contained both myc and SV40 sequences. This mouse developed a cranial bulge identified as a choroid plexus papilloma at 13 weeks and was subsequently sacrificed; tissue samples were taken for further analysis. Primary cell lines derived from these tissues contained both myc and SV40 DNA. No v-myc mRNA could be detected, although SV40 mRNA was present in all of the cell lines tested. T antigen also was expressed in all of the cell lines analyzed. These data suggest that SV40 expression was involved in the abnormalities of mouse M13 and was responsible for the transformed phenotype of the primary cell lines. Primary cell lines from this mouse were atypical in that the population rapidly became progressively more transformed with time in culture based on the following criteria: morphology, growth rate, and the ability to grow in soft agar and in serum-free medium. The data also suggest that factors present in the mouse regulated the ability of SV40 to oncogenically transform most cells and that in vitro culture of cells allowed them to escape those factors. PMID- 2985964 TI - Polyomavirus enhancer contains multiple redundant sequence elements that activate both DNA replication and gene expression. AB - Sequences that comprise the 244-base-pair polyomavirus enhancer region are also required in cis for viral DNA replication (Tyndall et al., Nucleic Acids Res. 9:6231-6250, 1981). We have studied the relationship between the sequences that activate replication and those that enhance transcription in two ways. One approach, recently described by de Villiers et al. (Nature [London], 312:242-246, 1984), in which the polyomavirus enhancer region was replaced with other viral or cellular transcriptional enhancers suggested that an enhancer function is required for polyomavirus DNA replication. The other approach, described in this paper, was to analyze a series of deletion mutants that functionally dissect the enhancer region and enabled us to localize four sequence elements in this region that are involved in the activation of replication. These elements, which have little sequence homology, are functionally redundant. Element A (nucleotides 5108 through 5130) was synthesized as a 26-mer with XhoI sticky ends, and one or more copies were introduced into a plasmid containing the origin of replication, but lacking the enhancer region. Whereas one copy of the 26-mer activated replication only to 2 to 5% of the wild-type level, two copies inserted in either orientation completely restored replication. We found that multiple copies of the 26-mer were also active as a transcriptional enhancer by measuring the beta-globin mRNA levels expressed from a plasmid that contained either the polyomavirus enhancer or one or more copies of the 26-mer inserted in a site 3' to the beta-globin gene. We observed a correlation between the number of inserted 26-mers and the level of beta-globin RNA expression. PMID- 2985965 TI - Myeloid cell transformation by ras-containing murine sarcoma viruses. AB - BALB and Harvey murine sarcoma viruses contain ras transforming genes capable of altering the proliferation and differentiation of cells within the erythroid and lymphoid lineages (W. D. Hankins and E. M. Scolnick, Cell 26:91-97, 1981; J. H. Pierce and S. A. Aaronson, J. Exp. Med. 156:873-887, 1982; E. M. Scolnick et al., Mol. Cell. Biol. 1:68-74). The present studies demonstrate hematopoietic targets of ras-containing viruses within the myeloid lineage. Diffuse colonies were induced by BALB or Harvey marine sarcoma virus infection of murine bone marrow cells. Generally, these colonies were made up of relatively mature macrophages which exhibited increased self-renewal capacity but eventually underwent terminal differentiation in culture. Cells from one BALB murine sarcoma virus-induced colony displayed phenotypic markers of more immature myelomonocytic cells. This colony, designated BAMC1, readily established as a continuous cell line and was highly malignant in vivo. Exposure of these cells to 12-O-tetradecanoylphorbol-13 acetate led to the induction of a more mature myeloid phenotype, which was associated with decreased growth potential in vitro and in vivo. The effects of the inducing agent were not mediated by an alteration in the level of expression of the ras-coded p21 transforming protein. Our present findings extend the spectrum of targets whose growth is altered by ras-containing retroviruses to cells at several stages of differentiation within each of the major hematopoietic lineages. PMID- 2985966 TI - A secreted factor and cyclic AMP jointly regulate cell-type-specific gene expression in Dictyostelium discoideum. AB - We are studying cell differentiation in Dictyostelium discoideum by examining the regulation of genes that are preferentially expressed in different cell types. A system has been established in which prestalk- and prespore-cell-specific genes are expressed in single cells in response to culture conditions. We confirm our previous results showing that cyclic AMP induces prestalk genes and now show that it is also required for prespore gene induction. The expression of both classes of genes is additionally dependent on the presence of a factor(s) secreted by developing cells which we call conditioned medium factor(s). An assay for conditioned medium factor(s) shows that it is detectable within 2.5 h after the onset of development. Conditioned medium factor(s) also promotes the expression of genes induced early in development, but has no detectable effect on the expression of actin genes and a gene expressed maximally in vegetative cells. In the presence of conditioned medium factor(s), exogenous cyclic AMP at the onset of starvation fails to induce the prespore and prestalk genes. The addition of cyclic AMP between 2 and 12 h of starvation results in rapid prestalk gene expression, whereas prespore genes are induced at an invarient time (approximately 18 h after the onset of starvation). These data suggest that cyclic AMP and conditioned medium factor(s) are sufficient for prestalk gene induction, whereas an additional parameter(s) is involved in the control of prespore gene induction. In contrast to several previous studies, we show that multicellularity is not essential for the expression of either prespore or prestalk genes. These data indicate that prespore and prestalk genes have cell type-specific as well as shared regulatory factors. PMID- 2985967 TI - Homologous recombination catalyzed by human cell extracts. AB - Two plasmids containing noncomplementing and nonreverting deletions in a bacterial phosphotransferase gene conferring resistance to neomycin (Neor) were incubated with human cell extracts, and the mixtures were used to transform recombination-deficient (recA-) Escherichia coli cells. We were able to obtain Neor colonies at a frequency of 2 X 10(-3). This frequency was 100 to 1,000 times higher than that obtained with no extracts. The removal of riboadenosine 5' triphosphate, Mg2+, or deoxynucleoside triphosphates from the reaction mixture severely reduced the yield of Neor colonies. Examination of plasmid DNA from the Neor colonies revealed that they resulted from gene conversion and reciprocal recombination. On the basis of these results, we conclude that mammalian somatic cells in culture have the enzymatic machinery to catalyze homologous recombination in vitro. PMID- 2985968 TI - Two integrated partial repeats of simian virus 40 together code for a super-T antigen. AB - We determined that the coding sequence for a 100-kilodalton super-T antigen found in Simian virus 40 mouse transformants spanned two separate partial repeats of the viral genome. The downstream repeat contained a complete Simian virus 40 large-T-antigen gene, whereas the upstream repeat was a truncated copy of the same gene. When the repeats were separated by subcloning, the capacity to code for the super-T antigen was lost. A small insertion or deletion in the origin control region which preceded the second repeat could also destroy the ability to code for the 100-kilodalton protein. Our data suggest that differential splicing between parts of two gene copies was responsible for the additional molecular weight of this super-T antigen. PMID- 2985969 TI - Two distinct families of human and bovine interferon-alpha genes are coordinately expressed and encode functional polypeptides. AB - The classical human interferon-alpha (HuIFN-alpha) gene family is estimated to consist of 15 or more nonallelic members which encode proteins sharing greater than 77% amino acid sequence homology. Low-stringency hybridization with a HuIFN alpha cDNA probe permitted the isolation of two distinct classes of bovine IFN alpha genes. The first subfamily (class I) is more closely related to the known HuIFN-alpha genes than to the second subfamily (class II) of bovine IFN-alpha genes. Extensive analysis of the human genome has revealed a HuIFN-alpha gene subfamily corresponding to the class II bovine IFN-alpha genes. The class I human and bovine IFN-alpha genes encode mature IFN polypeptides of 165 to 166 amino acids, whereas the class II IFN-alpha genes encode 172 amino acid proteins. Expression in Escherichia coli of members of both gene subfamilies results in polypeptides having potent antiviral activity. In contrast to previous studies which found no evidence of class II IFN-alpha protein or mRNA expression, we demonstrate that the class I and class II IFN-alpha genes are coordinately induced in response to viral infection. PMID- 2985971 TI - Rearrangements in the long terminal repeat of extra mouse mammary tumor proviruses in T-cell leukemias of mouse strain GR result in a novel enhancer-like structure. AB - Male GR mice develop T-cell leukemia at low frequency late in life. These leukemia cells invariably contain large amounts of mouse mammary tumor virus (MMTV) RNA and MMTV proteins and have extra MMTV proviruses integrated in their DNA. We show here that the extra MMTV proviruses are all derived from the endogenous MMTV provirus associated with the Mtv-2 locus and that the T-cell leukemias are clonal with respect to the acquired MMTV proviruses. The extra MMTV proviruses in six transplantable T-cell leukemia lines studied had rearranged, shortened long terminal repeats (LTRs); each T-cell leukemia, however, had a different LTR rearrangement within its extra MMTV provirus. The alteration within the extra LTRs of T-cell leukemia line 42 involved deletion of 453 nucleotides and generation of a tandem repeat region consisting of regions flanking the deletion. This alteration generated a sequence similar to the adenovirus enhancer core sequence. The viral RNAs in the T-cell leukemias contained corresponding alterations in their U3 regions. These results demonstrate that expression of MMTV in T-cell leukemias of GR mice may be the consequence of the generation of a novel enhancer, which could also stimulate expression of any adjacent cellular oncogene. PMID- 2985970 TI - Transcription from the polyoma late promoter in cells stably transformed by chimeric plasmids. AB - We have examined the expression of chimeric plasmids containing coding sequences for the herpes simplex virus thymidine kinase (tk) gene or the Tn5 gene for neomycin resistance (neo) linked to the late promoter of polyoma DNA. Although polyoma late genes are generally not expressed in transformed cells containing only integrated viral DNA molecules, rat tk- or wild-type cells transfected with the tk- or neo-containing plasmids were capable of growing in medium containing either hypoxanthine-aminopterin-thymidine or G418, respectively, under conditions nonpermissive for extrachromosomal DNA replication, indicating that the tk or neo genes were fully expressed. Moreover, cells were capable of growth in either hypoxanthine-aminopterin-thymidine or G418, even in the absence of direct selection for this activity. Northern analysis indicated steady-state levels of tk or neo transcripts that approximated the levels of polyoma early transcripts. S1 analysis showed that these transcripts initiated within the late promoter of polyoma and that their 5' ends mapped at positions similar or identical to those utilized during late lytic infection. The effect of substitution of polyadenylation signals was examined. Although plasmids containing the polyoma early polyadenylation signal were more efficient in conferring to cells a stable G418-resistant phenotype than similar constructions using the late signal, both signals were found to be effectively utilized. This indicates that the inability to detect late transcripts in polyoma-transformed cells in the absence of free viral DNA production is not an effect of inefficient mRNA cleavage or polyadenylation. Our results suggest that late gene expression in integrated polyoma genomes is not regulated at the level of message initiation but, most likely, through posttranscriptional events. PMID- 2985973 TI - A transforming activity not detected by DNA transfer to NIH 3T3 cells is detected by JB6 mouse epidermal cells. AB - Transfection of four different mouse epidermal tumor cell DNAs into NIH 3T3 cells yielded neither morphologically altered foci nor anchorage independence. However, promotion-sensitive, but not promotion-insensitive, JB6 mouse epidermal cell lines were permissive for the expression of anchorage independence after transfection of DNA from three of these tumor cell lines. This transforming activity and the promotion-sensitive activity that confers sensitivity to promotion of transformation show differences in restriction enzyme sensitivity. In view of this difference and the differences in both recipient cells and 12-O tetradecanoyl-phorbol-13-acetate dependence of expression, it appears that the transforming activity and the promotion-sensitive activity are specified by different genes. The JB6 promotion-sensitive cell lines may be useful for detecting and cloning transforming genes that escape detection in the NIH 3T3 cell focus assay. PMID- 2985974 TI - Genetic mapping of a cellular DNA region involved in induction of thymic lymphomas (Mlvi-1) to mouse chromosome 15. AB - Mlvi-1 defines a genetic locus representing a common domain for proviral DNA integration in Moloney murine leukemia virus-induced rat thymic lymphomas. Cellular sequences homologous to Mlvi-1 are present in mouse DNA, and we have used hamster-mouse somatic cell hybrids to chromosomally map Mlvi-1 in the mouse genome. Results demonstrated that Mlvi-1 maps to mouse chromosome 15 and that it is distinct from the Mlvi-2 integration region and from the cellular oncogenes c myc and c-sis, which also map to this chromosome. Therefore, Mlvi-1 may contain novel sequences involved in the establishment and maintenance of virus-induced murine tumors, many of which contain abnormalities of chromosome 15. PMID- 2985972 TI - Circular and linear simian virus 40 DNAs differ in recombination. AB - Linear forms of simian virus 40 (SV40) DNA, when added to transfection mixtures containing circular SV40 and phi X174 RFI DNAs, enhanced the frequency of SV40/phi X174 recombination, as measured by infectious center in situ plaque hybridization in monkey BSC-1 cells. The sequences required for the enhancement of recombination by linear DNA reside within the SV40 replication origin/regulatory region (nucleotides 5,171 to 5,243/0 to 128). Linearization of phi X174 RFI DNA did not increase the recombination frequency. The SV40/phi X174 recombinant structures arising from transfections supplemented with linear forms of origin-containing SV40 DNA contained phi X174 DNA sequences interspersed within tandem head-to-tail repeats derived from the recombination-enhancing linear DNA. Evidence is presented that the tandem repeats are not formed by homologous recombination and that linear forms of SV40 DNA must compete with circular SV40 DNA for the available T antigen to enhance recombination. We propose that the enhancement of recombination by linear SV40 DNA results from the entry of that DNA into a rolling circle type of replication pathway which generates highly recombinogenic intermediates. PMID- 2985975 TI - Drosophila P element-enhanced transfection in mammalian cells. AB - We constructed a gene transfer vector containing the herpes simplex virus type 1 thymidine kinase (TK) gene flanked by Drosophila P element terminal repeats (W. R. Engels, Annu. Rev. Genet. 17:315-344). This vector was introduced into mouse LTK- cells and enhanced the frequency of stable transformation to the TK+ phenotype by approximately 50-fold relative to a similar plasmid lacking the P element terminal repeats. PMID- 2985976 TI - Cystic islet cell tumor: diagnosis and surgical management. PMID- 2985977 TI - Hepatocellular carcinoma presenting as sphenoid sinus metastasis. PMID- 2985978 TI - A loss of uvrA function decreases the induction of the SOS functions recA and umuC by mitomycin C in Escherichia coli. AB - We have studied the levels of recA and umuC protein synthesis in Escherichia coli as a probe for regulatory and mechanistic events involved in mitomycin C mutagenesis. Both RecA and UmuC protein induction were greatly stimulated by mitomycin C in the wild-type strain, reached a peak at about 60 min for the recA gene, and at 90 min for the umuC gene, respectively, and maintained a plateau. The induction was blocked by recA and lexA(Ind-) mutations that conferred no mutagenesis on the cell. Mutation affecting uvrA protein markedly decreased induction of the recA gene as well as the umuC gene by mitomycin C. The results established that UvrA protein is involved in the induction of recA and umuC, and account, at least in part, for the mitomycin C nonmutability of uvrA mutants. PMID- 2985979 TI - An assessment of the importance of error-prone repair and point mutations to forward mutation to L-azetidine-2-carboxylic acid resistance in Escherichia coli. AB - By comparison of E. coli WP2 with CM891 (uvrA- pKM101) we found that pKM101 plasmid and uvrA- mutation considerably enhanced both spontaneous and chemically induced reversion at the trp locus. However, little or no increase was observed for forward mutation at the A2C locus. Furthermore, mutation frequency decline was considerably greater for trp reversion than for mutation to A2Cr. Thus neither error-prone repair nor point mutation seemed likely to be the major mechanism for forward mutation at the A2C locus. Results for spontaneous mutation of recA-, polA- and gyrA- strains showed that polA- and gyrA- gave good increases in forward mutation but not in reversion. It was inferred that deletion, transposition and/or larger chromosomal effects rather than point mutation were mainly responsible for most forward mutation. PMID- 2985980 TI - Influence of Neurospora endonuclease on trenimon-induced structural chromosomal aberrations and sister-chromatid exchanges in human peripheral lymphocytes and Chinese hamster ovary cells. AB - Human peripheral lymphocytes and Chinese hamster ovary cells were treated in the G1 phase of the cell cycle with the trifunctional alkylating agent trenimon (TRN) and post-treated with a single-strand specific endonuclease from Neurospora crassa (NE). TRN induces chromosomal aberrations of the chromatid type (CA) and sister-chromatid exchanges (SCE). NE post-treatment leads to an elevation of the frequencies of CA but not of SCEs. This indicates that TRN induced CA are the result of DNA double-strand breaks and that the SCEs originate from other types of lesions, most probably base damage. PMID- 2985981 TI - Nuclear and mitochondrial DNA of the primate malarial parasite Plasmodium knowlesi. AB - Restriction analyses and DNA/DNA hybridisation of parasite DNA isolated from monkeys infected with the malarial parasite Plasmodium knowlesi has permitted unambiguous identification of the nuclear DNA of this species. Its (G+C) content, as determined by estimations of buoyant density as well as by direct analysis, is about 38%, essentially indistinguishable from that of its primate laboratory host, and grossly different from that of the major human malaria parasite, P. falciparum, which has a (G+C) content of approx. 19%. In addition, gradient fractionation of total P. knowlesi DNA revealed a minor DNA component (approx. 1% of the total) with a (G+C) content of about 19%. This DNA comprises covalently closed circular molecules which have a contour length about 11.6 microns, carry a small cruciform structure, and are thought to originate in the parasite's mitochondria. PMID- 2985982 TI - Dolichol kinase in Ascaris suum and Onchocerca volvulus. AB - Dolichol kinase was demonstrated in the microsomal fraction of Ascaris suum and Onchocerca volvulus. The enzyme from nematodes exhibited specificity for CTP as phosphoryl donor and was found to be inhibited by the reaction product CDP. Enzyme activity was optimal at pH 7.4, in the temperature range between 30 degrees and 37 degrees C, and in the presence of 0.5% Triton X-100. In addition, the enzyme was found to depend on divalent cations for activity; Mg2+ being more effective than Mn2+ and Ca2+. The dolichol kinase from both nematodes was shown to be independent of Ca2+-calmodulin for activity. The apparent Km values for dolichol were determined to be 7.5 and 9.0 micrograms ml-1 for the enzyme from A. suum and O. volvulus, respectively. Those for CTP were estimated to be 0.85 and 0.75 mM, respectively. PMID- 2985983 TI - The presence of alpha-glycerophosphate dehydrogenase (NAD+-linked) and adenylate kinase as core and integral membrane enzymes respectively in the glycosomes of Trypanosoma rhodesiense. AB - A subcellular fraction enriched 12 times in glycosomes (NAD+-linked alpha glycerophosphate dehydrogenase) and devoid of detectable contamination from other subcellular components, was prepared from bloodstream Trypanosoma rhodesiense. Using a method employing exposure to toluene as a means of studying normally latent glycosomal enzymes, and phospholipase A2 as a membrane probe, the association of adenylate kinase and alpha-glycerophosphate dehydrogenase with the glycosome was studied. The normally latent glycerophosphate dehydrogenase (NAD+ linked), it is proposed, is an intraglycosomal enzyme having no membrane association, but bound to the core by weak ionic linkages. As such it is possible to release the enzyme from permeable (toluene treated) glycosomes using Cl-, with a resulting 4-fold increase in the Km for dihydroxyacetone phosphate. The presence of Cl- also stimulates an increase in specific activity, but this is observed before any release of enzyme. In contrast adenylate kinase, a non-latent glycosomal enzyme, is clearly membrane associated, the use of phospholipase A2 revealing an absolute dependence on phospholipid for activity. Restoration of activity appears to specifically require phosphatidyl choline and to be co operative in nature (nH = 1.56). It is proposed that adenylate kinase is an integral glycosomal membrane enzyme, probably affecting the control of intra glycosomal ADP/ATP levels. PMID- 2985984 TI - ADP-ribosyl transferase activity in Trypanosoma brucei. AB - Nuclear adenosine diphosphoribosyl transferase (ADPRT) catalyses the covalent modification of chromatin proteins by (ADP-ribose)n. This activity, which is entirely dependent on DNA containing strand breaks, is required for efficient DNA excision repair possibly because it regulates DNA ligation. ADPRT activity is also required for cytodifferentiation in a number of different cell types. We report here the presence of ADPRT activity in the blood-stream form of Trypanosoma brucei and its activation by DNA strand breaks formed by exposure to, either exogenously supplied deoxyribonuclease I, or treatment with the methylating agent, dimethylsulphate. 3-Aminobenzamide, but not its chemical analogue 3-aminobenzoic acid, is a competitive inhibitor of ADPRT activity in T. brucei. Intact trypanosomes are readily permeable to this competitive inhibitor of ADPRT activity. PMID- 2985985 TI - Further analysis of intraspecific variation in Trypanosoma brucei using restriction site polymorphisms in the maxi-circle of kinetoplast DNA. AB - We have compared the maxi-circle kinetoplast DNA of 21 Trypanosoma brucei sp. stocks by analysis of restriction sites for nine restriction endonucleases. The analysis shows most of these stocks to have a maxi-circle sequence similar to that of 11 previously analysed stocks, with a difference of less than 3% between any two stocks. However, seven stocks stand out from the rest with at least two sites lost or gained for six of the nine restriction enzymes used. These seven distinctive stocks fall into two groups with some shared and some unique polymorphisms. One group had already been designated the kiboko group on the basis of isoenzyme patterns, but the relationship between nuclear markers and maxi-circle type is less clear-cut for the other group, designated sindo. Both groups seem to be in a wild animal-tsetse fly transmission cycle, with occasional infections in domestic stock, and may be reproductively isolated from the main T. brucei sp. population. The existence of the kiboko and sindo sub-groups shows that the maxi-circle is not shielded from evolutionary change. The lack of difference observed between the maxi-circles of the majority of T. brucei sp. stocks, including the gambiense and rhodesiense variants, must therefore reflect their close homology. Two geographical trends occur in T. brucei as a whole: (a) a trend in maxi-circle size, with increasing length of the variable region from West to East Africa, and (b) a greater frequency of certain restriction enzyme polymorphisms in East African stocks as compared to West African stocks. PMID- 2985987 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 19-1985. A 46-year-old woman with obstruction of the right pulmonary artery. PMID- 2985986 TI - Effect of dietary enrichment with eicosapentaenoic and docosahexaenoic acids on in vitro neutrophil and monocyte leukotriene generation and neutrophil function. AB - The effects of dietary fish-oil fatty acids on the function of the 5-lipoxygenase pathway of peripheral-blood polymorphonuclear leukocytes and monocytes were determined in seven normal subjects who supplemented their usual diet for six weeks with daily doses of triglycerides containing 3.2 g of eicosapentaenoic acid and 2.2 g of docosahexaenoic acid. The diet increased the eicosapentaenoic acid content in neutrophils and monocytes more than sevenfold, without changing the quantities of arachidonic acid and docosahexaenoic acid. When the neutrophils were activated, the release of [3H]arachidonic acid and its labeled metabolites was reduced by a mean of 37 per cent, and the maximum generation of three products of the 5-lipoxygenase pathway was reduced by more than 48 per cent. The ionophore-induced release of [3H]arachidonic acid and its labeled metabolites from monocytes in monolayers was reduced by a mean of 39 per cent, and the generation of leukotriene B4 by 58 per cent. The adherence of neutrophils to bovine endothelial-cell monolayers pretreated with leukotriene B4 was inhibited completely, and their average chemotactic response to leukotriene B4 was inhibited by 70 per cent, as compared with values determined before the diet was begun and six weeks after its discontinuation. We conclude that diets enriched with fish-oil-derived fatty acids may have antiinflammatory effects by inhibiting the 5-lipoxygenase pathway in neutrophils and monocytes and inhibiting the leukotriene B4-mediated functions of neutrophils. PMID- 2985988 TI - Lack of correlation between promiscuity and seropositivity to HTLV-III from a low incidence area for AIDS. PMID- 2985989 TI - Lack of evidence of HTLV-III endemicity in southern Africa. PMID- 2985990 TI - Serum lactate dehydrogenase and bone marrow involvement in small-cell carcinoma of the lung. PMID- 2985991 TI - Genotyping with DNA probes in combined immunodeficiency syndrome with defective expression of HLA. AB - We performed HLA genotyping by using restriction-enzyme fragments hybridized with specific HLA probes instead of traditional immunologic methods in two patients whose lymphocytes expressed so few HLA antigens on the cell surface that serologic methods failed. Segregation of restriction-fragment-length polymorphism permitted identification of the genotypes. In addition, known correlations between serologically determined antigens and restriction-fragment-length polymorphism were confirmed. We applied this approach in making therapeutic decisions regarding bone marrow transplantation. PMID- 2985992 TI - Transfusion-associated acquired immunodeficiency syndrome. Evidence for persistent infection in blood donors. AB - To investigate whether infection with human T-cell lymphotropic virus/lymphadenopathy-associated virus (HTLV-III/LAV) may be persistent in asymptomatic persons and to correlate infection with seropositivity, we performed virologic and serologic studies in 25 of 30 persons who were identified as being at high risk for the acquired immunodeficiency syndrome (AIDS) and who had donated blood to patients who later contracted transfusion-associated AIDS. High risk donors were those who belonged to a high-risk population, had AIDS or a closely related condition, or had a low ratio of helper to suppressor T lymphocytes. We performed similar studies in 6 of the 24 patients with AIDS who had received donations from this group. HTLV-III/LAV was isolated from 22 of the 25 donors, between 12 and 52 months (mean, 28) after they had donated blood, and from all 6 recipients, between 14 and 37 months (mean, 26) after they had received blood. Of the 22 virus-positive donors, 2 have contracted AIDS, 5 have generalized lymphadenopathy, and 15 (68 per cent) remain asymptomatic. Antibodies to HTLV-III/LAV were detectable by the enzyme-linked immunosorbent assay in serum samples obtained from each person at the time the virus was isolated. We conclude that infection with HTLV-III/LAV may be persistent and asymptomatic for years. This demonstration that viremic patients may be asymptomatic supports the use of serologic screening of donated blood to supplement current procedures for the prevention of transfusion-associated AIDS. PMID- 2985993 TI - Association of Epstein-Barr virus with thymic carcinoma. PMID- 2985994 TI - "Lymphoepithelioma-like" thymic carcinoma: another tumor related to Epstein-Barr virus? PMID- 2985995 TI - In vivo activated T lymphocytes in the peripheral blood and cerebrospinal fluid of patients with multiple sclerosis. AB - We found an increase in peripheral-blood lymphocytes bearing the T-cell-specific activation antigen Ta1 in 20 of 35 patients with progressive multiple sclerosis, 4 of 18 patients with stable or improving multiple sclerosis, 1 of 17 patients with other neurologic diseases, and 1 of 14 normal controls (P less than 0.0002, Fisher's exact test). No increases in two other markers of T-cell activation, T113 and the interleukin-2 receptor, were found. In the cerebrospinal fluid, patients with progressive multiple sclerosis (pleocytosis, 3.9 +/- 1.6 cells per cubic millimeter) had 42 +/- 3.0 per cent Ta1+ cells. In contrast, patients with other inflammatory central nervous system diseases (36 +/- 13 cells per cubic millimeter) had 9.6 +/- 1.8 per cent Ta1+ cells (P less than 0.01). In patients with other neurologic diseases without inflammation (0.7 +/- 0.16 cells per cubic millimeter), the percentage of Ta1+ cells was equivalent to that in patients with multiple sclerosis (39 +/- 5.4 per cent), although the absolute number was lower. There was a positive correlation between the presence of Ta1+ cells in the spinal fluid and blood of patients with other neurologic diseases, but not patients with multiple sclerosis. Less than 1 per cent of lymphocytes from the spinal fluid of patients with multiple sclerosis expressed interleukin-2 receptors, as compared with 9.8 per cent of cells from subjects with other inflammatory neurologic diseases (P less than 0.01). These results suggest that the T cells in the spinal fluid of patients with multiple sclerosis may be activated by a different mechanism or in a different temporal sequence from that in patients with other nervous system diseases. Furthermore, the increase in Ta1+ cells in the peripheral blood of patients with multiple sclerosis demonstrates systemic immune activation in the disease; monitoring such cells may provide an objective measure of abnormal immunologic activity. PMID- 2985996 TI - Inferior petrosal venous sinus sampling in lateralization of ACTH-secreting pituitary microadenomas. PMID- 2985997 TI - Antilymphocyte antibodies and seropositivity for retroviruses in groups at high risk for AIDS. PMID- 2985998 TI - Absence of immunosuppression in healthy subjects from eastern Zaire who are positive for HTLV-III antibody. PMID- 2986000 TI - Study of stroma in scirrhous gastric carcinoma. PMID- 2985999 TI - Antifungal activity of ovotransferrin towards genus Candida. AB - The inhibiting activity of ovotransferrin was tested towards different species belonging to genus Candida. Of one hundred strains tested, only C. krusei showed a noticeable resistance, while the other species appeared to be more sensitive than bacteria to the action of ovotransferrin. The influence of anions, such as bicarbonate and citrate, on the inhibiting activity of ovotransferrin was also investigated. Moreover it was observed that iron saturated ovotransferrin retained its activity, thus suggesting an interaction between the protein and Candida cells. PMID- 2986001 TI - Relationship between interstitia and prognosis of gastric carcinoma. PMID- 2986002 TI - Opposite motivational effects of endogenous opioids in brain and periphery. AB - Many psychoactive drugs, including the opiates, have been shown to have paradoxical reinforcing effects. Opiates produce positive reinforcing effects when they are paired with visual and textural environmental stimuli in rats, yet, at similar doses and over the same routes of administration, produce aversive effects, as shown when they are paired with taste stimuli. Similarly, in human, the positive reinforcing effects of opiates are well known to addicts and recreational drug users, yet patients receiving opiates as analgesics often report nauseous reactions. At present there is no evidence to differentiate between the neural substrates that mediate these opposite motivational effects. We now report an initial step in the resolution of this paradox by demonstrating that endogenous and exogenous opioids produce positive reinforcing effects through an action on brain opiate receptors, and aversive effects through an action on peripheral opiate receptors (especially in the gut). PMID- 2986003 TI - Role of guanine nucleotide binding protein in the activation of polyphosphoinositide phosphodiesterase. AB - Interaction of ligands with 'Ca2+-mobilizing' receptors is thought to result in the generation of two second messengers, inositol trisphosphate and diacylglycerol, from a common substrate, phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) (refs 1, 2), a component of plasma membranes. It is not known how the occupation of such receptors is translated into the activation of the catalytic unit polyphosphoinositide (PPI) phosphodiesterase, and then to cellular activation, but our recent experiments suggest that GTP regulatory proteins may be involved. In mast cells, non-hydrolysable analogues of GTP introduced and then trapped in the cytosol are able to substitute for external ligands in inducing exocytosis, a well-defined Ca2+-dependent process, suggesting that guanine nucleotide regulatory proteins may act by stimulating the catalytic activity of the PPI phosphodiesterase. We now provide evidence that mast cell secretion is inhibited by internalized neomycin, a compound known to interact with PPI. We also show that the PPI phosphodiesterase of human neutrophil plasma membranes can be activated simply by adding GTP analogues in the presence of concentrations of Ca2+ that pertain in unstimulated cells. These findings strongly support the idea that the coupling factor linking receptor and PPI phosphodiesterase is a guanine nucleotide binding protein analogous to those involved in the activation and inhibition of adenylate cyclase. PMID- 2986004 TI - Increased neurovirulence associated with a single nucleotide change in a noncoding region of the Sabin type 3 poliovaccine genome. AB - Most of the small number of cases of poliomyelitis which occur in countries where Sabin's attenuated poliovirus vaccines are used are temporally associated with administration of vaccine and involve polioviruses of types 2 and 3 (ref. 1). Recent studies have provided convincing evidence that the Sabin type 2 and 3 viruses themselves may revert to a neurovirulent phenotype on passage in man. We report here that a point mutation in the 5' noncoding region of the genome of the poliovirus type 3 vaccine consistently reverts to wild type in strains isolated from cases of vaccine-associated poliomyelitis. Virus with this change is rapidly selected on passage through the human gastrointestinal tract. The change is associated with a demonstrable increase in the neurovirulence of the virus. PMID- 2986005 TI - Suppression of leukaemia virus pathogenicity by polyoma virus enhancers. AB - The long terminal repeats (LTRs) of retroviruses contain sequences necessary for the initiation and termination of retroviral transcription. These sequences include promoter elements, transcriptional termination signals and transcriptional enhancer elements. The enhancer elements of Moloney murine leukaemia virus (M-MuLV) are localized in a tandemly repeated region (approximately 75 base pairs (bp) long), which lies 5' to the CAT and TATA promoter elements in the U3 region of the LTR (see Fig. 1). We have shown that the tandem repeats are required both for LTR promoter activity, as measured by transient expression assays, and for biological activity, as measured by production of infectious virus. Furthermore, they can be replaced by transcriptional enhancers from the F101 host-range mutant of polyoma virus without loss of function. We report here that the addition of the polyoma (PyF101) enhancers to the M-MuLV LTRs (either with or without the M-MuLV tandem repeats) results in complete loss of viral leukaemogenicity, even though the virus can replicate to high titres in tissue culture fibroblasts and can establish infection in animals. PMID- 2986006 TI - ATP-dependent specific binding of Tn3 transposase to Tn3 inverted repeats. AB - Transposons are discrete segments of DNA which are capable of moving from one site in a genome to many different sites. Tn3 is a prokaryotic transposon which is 4,957 base pairs (bp) long and encodes a transposase protein which is essential for transposition. We report here a simple method for purifying Tn3 transposase and demonstrate that the transposase protein binds specifically to the ends of the Tn3 transposon in an ATP-dependent manner. The transposase protein binds to linear double-stranded DNA both nonspecifically and specifically; the nonspecific DNA binding activity is sensitive to challenge with heparin. Site-specific DNA binding to the ends (inverted repeats) of Tn3 is observed only when binding is performed in the presence of ATP; this ATP dependent site-specific DNA binding activity is resistant to heparin challenge. Our results indicate that ATP qualitatively alters the DNA binding activity of the transposase protein so that the protein is able to bind specifically to the ends of the Tn3 transposon. PMID- 2986007 TI - Name for AIDS virus. PMID- 2986008 TI - Renal cytochrome P450-related arachidonate metabolite inhibits (Na+ + K+)ATPase. AB - The function of the nephron, the anatomical unit of the kidney, is segmented; at least 12 segments have been identified that differ in their morphology, transport properties and hormonal responsiveness. The medullary portion of the thick ascending limb of the loop of Henle (mTALH) has one of the highest concentrations of (Na+ + K+)ATPase found in mammalian tissues, reflecting the importance of this nephron segment in the regulation of extracellular fluid volume, as active sodium transport is driven by (Na+ + K+)ATPase. Here, in cells derived primarily from the mTALH of the outer medulla of rabbit kidney, we have identified a cytochrome P450-dependent monooxygenase system which metabolizes arachidonic acid to two biologically active oxygenated products; one of the products inhibits (Na+ + K+)ATPase and the other relaxes blood vessels. We report that formation of these oxygenated arachidonate metabolites is stimulated by arginine vasopressin (AVP) and salmon calcitonin (SCT). PMID- 2986009 TI - Identification of the human analogue of a regulator that induces differentiation in murine leukaemic cells. AB - We have recently purified murine granulocyte colony-stimulating factor (G-CSF), a regulatory glycoprotein which stimulates granulocyte colony formation from committed murine precursor cells in semi-solid agar cultures. G-CSF is one of a family of colony-stimulating factors that regulate the growth and differentiation of granulocytes and macrophages. While the other murine CSFs (granulocyte macrophage (GM)-CSF, macrophage (M)-CSF and multi-CSF) show little or no differentiation-inducing activity on murine myelomonocytic leukaemia cell lines, G-CSF (or MGI-2(6)) is able to induce the production of terminally differentiated cells from WEHI-3B and other myeloid leukaemia cell lines. More importantly, G CSF-containing materials suppress the self-renewal of myeloid leukaemia stem cells in vitro and the leukaemogenicity of treated myeloid leukaemic cells in vivo. It is important to identify the human analogue of murine G-CSF so that its effectiveness on human myeloid leukaemia cells can be assessed. Here we show that an analogue of G-CSF does exist among the CSFs produced by human cells and that the murine and human molecules show almost complete biological and receptor binding cross-reactivities to normal and leukaemic murine or human cells. The human G-CSF analogue is identified as a species of CSF that we have previously described as CSF-beta. PMID- 2986010 TI - Structural analysis of murine genes containing homoeo box sequences and their expression in embryonal carcinoma cells. AB - The presence of homoeo box sequences in the genomes of vertebrates has suggested that these metazoans possess the equivalent of the homoeotic genes that have a key role in regulating the development of the fruitfly Drosophila melanogaster. We report here that a novel murine homoeo box-containing gene is expressed in embryonal carcinoma stem cells. Transcripts of the sequences that flank the homoeo box of this gene are found in these cells before and after induced differentiation, whereas a specific transcript that seems to contain the homoeo sequence is only present after differentiation. PMID- 2986011 TI - Genetic mapping and diagnosis of haemophilia A achieved through a BclI polymorphism in the factor VIII gene. AB - Haemophilia A is the most common inherited bleeding disorder in man, affecting approximately 1 male in 10,000. The disease is caused by a deficiency in the gene for factor VIII, a component of the intrinsic coagulation pathway. Due to the broad range of clotting activity in normal and heterozygous females, it is often difficult to confirm the status of women at risk for carrying the disease. A genetic marker in the form of a restriction fragment length polymorphism (RFLP) within or tightly linked to the factor VIII gene would serve as a tag for the haemophilia gene, thus allowing both accurate carrier detection and improved, earlier prenatal diagnosis by chorionic villi sampling. The recent isolation of the factor VIII gene has allowed a search for RFLPs within the gene, and we report here the identification of a common polymorphism within the factor VIII gene, revealed by the restriction enzyme BclI, which can be used diagnostically in about 42% of all families. Although the disease haemophilia A has been mapped to the distal portion of Xq, the BclI RFLP makes possible higher-resolution genetic linkage mapping with respect to other polymorphic markers on this portion of the X chromosome. We have established close linkage of the factor VIII gene to several useful RFLP markers, including the highly informative marker St14. These markers should also be useful for prenatal diagnosis of haemophilia A and for detection of its carriers. PMID- 2986012 TI - Most classical Mus musculus domesticus laboratory mouse strains carry a Mus musculus musculus Y chromosome. AB - Restriction fragment length polymorphisms (RFLPs) of maternally inherited mitochondrial DNA have revealed that genetic exchanges can occur between populations, subspecies or even species. From the point of view of population genetics the mammalian Y chromosome represents a genomic analogue of the mitochondrion; it is inherited only paternally and remains perpetually monosomic, showing little meiotic recombination with other chromosomes. Using a Y-specific genomic DNA probe obtained from a flow-sorted mouse Y-chromosome library, we have examined the RFLPs in 10 newly established mouse lines of the European semispecies Mus musculus domesticus and Mus musculus musculus, and identified two variant forms of the Y chromosome, each of which is characteristic of one of the semispecies. As reported here, probing the DNA from nine established inbred laboratory strains reveals that the strain SJL carries a M.m. domesticus type Y whereas, surprisingly, the Y chromosomes of the other eight strains are of M.m. musculus origin. Hence, these strains cannot be regarded as archetypes of M.m. domesticus as suggested by protein polymorphisms and mitochrondrial DNA sequence data, but rather as genetic hybrids between the two semispecies. PMID- 2986013 TI - Simian virus 40 enhancer increases number of RNA polymerase II molecules on linked DNA. AB - Since the discovery of transcription enhancer sequences in the genomes of the DNA viruses simian virus 40(SV40) and polyoma virus, these elements have been shown to play an important part in the control of both viral and cellular gene expression. Enhancer elements act in cis to increase the amount of RNA produced from linked genes in a manner largely independent of distance and orientation. The mechanisms by which enhancers act are not understood; in particular, it is not known whether the enhancer-dependent increase in the level of stable RNA reflects an increase in the rate of transcription. To address this question, we have used an in vitro nuclear transcription assay to examine the effect of the SV40 enhancer on transcription of cloned human beta-globin genes transiently introduced into HeLa cells. We show here that the SV40 enhancer acts at least in part to increase the number of RNA polymerase II molecules transcribing the linked gene. PMID- 2986014 TI - Simian virus 40 enhancer increases RNA polymerase density within the linked gene. AB - Enhancers are regulatory DNA elements, usually about 200 base pairs (bp) long, which are able to stimulate transcription of linked genes in eukaryotic cells. This activation can be exerted over large distances, and from a position 5' or 3' to the gene. Enhancers have been identified in viral genomes and cellular genes. Using a transient expression assay, we have analysed transcription of the rabbit beta-globin gene and the thymidine kinase gene from herpes simplex virus with and without a simian virus 40 (SV40) enhancer. S1 nuclease mapping shows a high level of specific transcripts when the genes are linked to the enhancer. To determine whether this increased number of transcripts is due to a higher transcription rate, or perhaps to a shift from nonspecific to specific initiation, we have performed run-on transcription assays with isolated nuclei. Our results, presented here, demonstrate that the SV40 enhancer increases the RNA polymerase density within the linked gene. Therefore, enhancers apparently increase the rate of transcription initiation without influencing the specificity of initiation. PMID- 2986015 TI - Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes. AB - Following the transfer into fertilized mouse eggs of recombinant genes composed of the upstream region of the rat insulin II gene linked to sequences coding for the large-T antigen of simian virus 40, large-T antigen is detected exclusively in the beta-cells of the endocrine pancreas of transgenic mice. The alpha- and delta-cells normally found in the islets of Langerhans are rare and disordered. Well-vascularized beta-cell tumours arise in mice harbouring and inheriting these hybrid oncogenes. PMID- 2986016 TI - An HTLV-III peptide produced by recombinant DNA is immunoreactive with sera from patients with AIDS. AB - Human T-cell lymphotropic retrovirus type III (HTLV-III), also called lymphadenopathy-associated virus (LAV), has been identified as the aetiological agent of acquired immune deficiency syndrome (AIDS). The sera of most patients with AIDS or AIDS-related complexes, and of asymptomatic individuals infected with HTLV-III, contain antibodies against antigens of HTLV-III. The characterization of these antibodies and their corresponding viral antigens is important not only for understanding immunity against HTLV-III and the pathology of AIDS, but also for the development of diagnostic methods and preventive vaccine for AIDS. Following the successful establishment of a long-term T-cell line permissive for HTLV-III replication, large quantities of virus have been produced, facilitating the purification of viral proteins and the development of mouse monoclonal antibodies against several viral antigens. More recently, the structure of HTLV-III proviral DNA has been elucidated. We now report the production, by genetic engineering methods, of a peptide encoded by a gene segment of HTLV-III. A 1.1-kilobase (kb) EcoRI DNA segment from an isolate of HTLV-III was inserted into a lpp and lac promoter-coupled expression vector, pIN III-ompA. Escherichia coli transformants of this plasmid produced a peptide of relative molecular mass (Mr) 15,000 (15K) which was strongly immunoreactive with anti-HTLV-III antibodies present in sera from AIDS patients. Lysates of the clones expressing this 15K peptide inhibited the reactivity of the p31 virion protein with AIDS sera, suggesting that it is a fragment of the viral p31 protein. The peptide reacted with sera from all 20 AIDS patients but none of the 8 normal controls tested. These results suggest that the peptide may be useful for detecting anti-HTLV-III antibodies in blood samples. PMID- 2986018 TI - [AIDS: a clinical diagnosis, hence one with pitfalls]. PMID- 2986017 TI - Autoradiographic localization of "peripheral-type" benzodiazepine binding sites in the rat brain, heart and kidney. AB - The localization of peripheral-type benzodiazepine binding sites has been accomplished using quantitative receptor autoradiography after labeling slide mounted tissue sections with [3H]-RO 5-4864. Specific [3H]-RO 5-4864 binding sites were observed in the choroid plexus and ependyma cells within the brain. Lower levels of specific binding were seen in areas corresponding to the glomerular layer of the olfactory bulb. Stereotaxic administration of the excitotoxin, ibotenic acid, into the piriform cortex produced a dramatic increase in binding to the peripheral-type site in this brain region. Binding in the kidney was associated with the ascending limb of the loop of Henle and the distal convoluted tubule, while in the heart a more diffuse binding was found throughout the ventricle wall. The localization of peripheral-type benzodiazepine binding sites to areas involved in ion transport gives additional support to previously suggested physiological roles for these sites. PMID- 2986019 TI - [Inhibition of H+/K+-ATPase in the parietal cell with omeprazole: a new principle of gastric acid inhibition]. PMID- 2986020 TI - Evidence for the cerebral uptake in vivo from two pools of glucose and the role of glucose-6-phosphatase in removing excess substrate from brain. AB - We propose the following scheme for cerebral uptake and overall metabolism of glucose in vivo: that brain selects from two pools of glucose anomers in arterial blood, that it takes up excess glucose, that glucose enters the brain tissue as glucose-6-phosphate through the actions of mutarotase and hexokinase, that some glucose-6-phosphate becomes metabolized to CO2 and some becomes incorporated into brain carbon pools, and that excess glucose-6-phosphate leaves brain through glucose-6-phosphatase and mutarotase activities. This results from our observations in arterio-venous studies for the determination of cerebral metabolism in humans in vivo that the cerebral uptake of [14C]glucose often appeared to differ from that of unlabeled glucose. With rapidly falling arterial radioactivity, unlabeled glucose uptake was more than [14C]glucose. With rising arterial radioactivity, [14C]glucose extraction exceeded unlabeled glucose. Studies with [14C]glucose-6-phosphate suggested that glucose-6-phosphatase in brain removes excess substrate by dephosphorylation. However, when arterial [14C]glucose increased slowly, [14C]glucose uptake varied considerably and the data resembled human cerebral metabolism of glucose anomers. An experiment employing [13C]glucose and NMR provided further support for our proposed scheme. PMID- 2986021 TI - Biochemical studies on rabbits with aluminium induced neurofilament accumulations. AB - The activities of acid phosphatase, hexosaminidase, beta-galactosidase, Mg2+ stimulated Na+K+ATPase, fumarase and ATP:citrate lyase were measured in grey matter of rabbit spinal cord 7-8 days after intra-ventricular or intra-cisternal injection of aluminium. RNA, DNA, and water content were measured in whole spinal cords. Choline acetyltransferase (CAT) and acetylcholinesterase were assayed in dorsal grey matter of the cord, which contained no aluminium-induced neurofilament accumulations (NFAs), and ventral grey matter, which had large numbers of such NFAs. CAT was also assayed in the hypoglossal nerve. None of these measures were consistently altered in the aluminium treated rabbits, although the activity of beta-galactosidase was increased in the NFA-free caudate nucleus of rabbits given aluminium intra-ventricularly, possibly due to the presence of phagocytes on the ventricular surface of the caudate. It is concluded that neither aluminium nor its induced NFAs has a gross effect on neuronal metabolism within 7-8 days. PMID- 2986022 TI - [Chemical embolization using estrogen for aneurysmal bone cyst of the skull: case report]. AB - A case of aneurysmal bone cyst of the fronto-parietal bone was reported. The patient was a 48-year-old female with the complaints of conjunctival hyperemia and exophthalmus of the right side. Right selective externography revealed both arteriovenous fistula between right middle meningeal artery and diploic vein, and abnormal venous pooling draining into right superior ophthalmic vein. These symptoms probably caused by the increased intraorbital venous pressure in the right side. Skull plain film and CT scan showed blow-out appearance of right fronto-parietal bone in accordance with the enlargement of diploic space. The bulged area and the rest of the affected part of the bone presented faintly coarse "soap bubble" pattern. And the affected bone, being radiolucent, had well demarcated borders which are distinct and smooth, with faint sclerosis. Both arteriovenous fistula and clinical symptom have disappeared completely by the continuous injection of conjugated estrogen into the right external carotid artery. This is the first case that proved chemical embolization using estrogen will effect a cure in the treatment of aneurysmal bone cyst. PMID- 2986024 TI - Diurnal variations in responsiveness of the hypothalamo-pituitary-adrenocortical axis of the rat. AB - Hypothalami, anterior pituitary gland segments and adrenal glands were removed from female Wistar-derived rats decapitated at various times of the day. Blood and tissue hormone concentrations were measured and the tissues challenged with appropriate stimuli in vitro. Both bioactive and immunoreactive corticotrophin releasing factor (CRF) content of the hypothalami were significantly higher in the evening than in the morning, as was the basal release of bioactive CRF in vitro. The response of the hypothalami to serotonin or acetylcholine added in vitro did not change with time of day. Basal bioactive and immunoreactive adrenocorticotrophin (ACTH) release from the anterior pituitary gland was significantly increased in the evening, as was the response to synthetic ovine CRF in vitro. Plasma ACTH concentrations in intact rats given crude CRF (hypothalamic extract) in vivo were higher in the evening at all times after injection tested, but this difference was markedly reduced in animals with mediobasal hypothalamic lesions. Corticosterone released basally from adrenal glands in vitro was significantly increased in the evening and the response to added ACTH 1-24 was slightly enhanced. For adrenal glands removed from lesioned rats, the pattern was reversed, corticosterone release in vitro being lower in the evening for all doses of ACTH added. Similarly in vivo, in intact rats given ACTH 1-24, plasma corticosterone concentrations and corticosterone release in vitro from adrenal glands removed after the injection were higher in the evening. After the placement of basal hypothalamic lesions, the situation was reversed, the response to ACTH administration in vivo being greater in the morning.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986023 TI - Cimetidine-induced prolactin release: possible involvement of the GABA-ergic system. AB - The mechanism of cimetidine-induced prolactin (PRL) release was studied. Intracarotid (i.a.) administration of 1 mg/kg of impromidine, the most specific H2 histamine agonist known, did not counteract the cimetidine-induced hypersecretion of PRL. Pre-treatment with benzodiazepines (diazepam or lorazepam, 3 mg/kg/i.a.) completely suppressed it. Administration of gamma-aminobutyric acid (GABA 5 mg/kg/i.a.) was also able to prevent it, but to a lesser extent than benzodiazepines. Simultaneous administration of doses of diazepam (1.5 mg/kg/i.a.) and GABA (3 mg/kg/i.a.) ineffective per se markedly blunted the increase of PRL by cimetidine. We conclude that cimetidine does not induce hypersecretion of PRL by its action on histamine H2 receptors, but through other pharmacological activities of the drug, such as perhaps interaction with the GABA ergic system in the pituitary. PMID- 2986025 TI - Effects of clonidine on blood pressure, noradrenaline, cortisol, growth hormone, and prolactin plasma levels in high and low intestinal tone subjects. AB - Systolic blood pressure (SBP), diastolic blood pressure (DBP), norepinephrine (NE), cortisol (CRT), growth hormone (GH), and prolactin (PRL) plasma levels were investigated in 46 normal subjects, 28 high intestinal tone (high IT) and 18 low intestinal tone (low IT), before and after the administration of a single intramuscular dose of clonidine (2.5 micrograms/kg). High IT subjects had lower mean values of DBP than low IT subjects, and basal NE was significantly greater in low IT than in high IT subjects. A negative correlation between NE and IT values was found for the high IT, but not for the low IT group, during the preclonidine periods. The drug reduced SBP in high IT, whereas it reduced SBP plus DBP and NE in low IT subjects. Clonidine induced significant reductions of CRT and increases of GH in both groups; furthermore, a slight but significant reduction of PRL was registered in high IT group. The drug also induced increase of distal colon tone in high IT subjects and suppressed phasic activity (waves) in low IT subjects. While a significant positive correlation was found between NE and DBP in low IT subjects during postclonidine periods, no correlation was found between the two parameters in high IT subjects. Other significant positive (+) and negative (-) correlations during postclonidine periods were: CRT/GH (-), CRT/PRL (+), and GH/PRL (-) in high IT subjects; NE/CRT (+), NE/GH (-), CRT/GH ( ), CRT/DBP (+), and GH/DBP (-) in low IT subjects. Finally, significant negative correlation was found between NE and distal colon tone during postclonidine periods in high IT subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986026 TI - A detailed examination of the in vivo and in vitro effects of ACTH on gonadotropin secretion in the adult rat. AB - These studies were designed to: (1) determine the effects of continuous infusion of synthetic ACTH(1-24) on postcastration changes in serum and pituitary LH, FSH and prolactin in the male rat; (2) assess the effects of adrenalectomy on the gonadotropin and prolactin response to ACTH, and (3) test the hypothesis that ACTH may directly (not via adrenal factors) alter gonadotropin secretion at the brain and/or pituitary level. Adult male rats were either orchidectomized (ORX) or orchidectomized-adrenalectomized (ORX-ADX), and were treated continuously for 6 days with ACTH(1-24) (10 micrograms/day) or saline using an osmotic minipump. Animals were killed on day 6 following castration. ACTH treatment reduced serum LH and prolactin levels in ORX rats to mean values +/- SE of 204 +/- 25 and 37 +/ 3 ng/ml respectively, compared to 366 +/- 72 and 62 +/- 7 ng/ml in saline treated ORX animals. Serum FSH concentrations were not altered by ACTH administration. Pituitary concentrations of LH and FSH, but not prolactin were enhanced by ACTH treatment. Adrenalectomy had no effect on serum and pituitary gonadotropin and prolactin levels, but abolished the effects of ACTH on these parameters. Central (intracerebroventricular) infusion of ACTH(1-24) (6 micrograms/day X 4 days) failed to alter the rise in serum LH in male rats following orchidectomy. Acute treatment with large doses of ACTH of perifused anterior pituitary glands from male rats and chronic treatment with ACTH of enzymatically dispersed anterior pituitary cells from female rats did not influence basal or GnRH-stimulated LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986027 TI - Modification of basal and GRF-stimulated cyclic AMP levels and growth hormone release by phospholipid metabolic enzyme inhibitors. AB - The relative importance of several phospholipid pathways in cyclic AMP (cAMP) metabolism and growth hormone (GH) release was determined by an indirect, pharmacological approach in cultured anterior pituitary cells. The diglyceride lipase inhibitor RHC-80267 (30-100 microM) had no significant effect on cAMP levels but markedly inhibited basal and growth hormone-releasing factor-(GRF) stimulated GH secretion. A phospholipase A2 inhibitor quinacrine (30 microM) increased cellular cAMP content while decreasing GH release. Indomethacin, which reduces cyclooxygenase activity, affected neither cAMP levels nor GRF-enhanced GH release; this drug (30-100 microM) did reduce basal GH release. The lipoxygenase inhibitors nordihydroguaiaretic acid and BW-755c both reduced basal and GRF stimulated GH release in a concentration-dependent manner. Both agents had various effects on cAMP levels. These results suggest that phospholipid metabolism, through both the cyclooxygenase and lipoxygenase pathways, contributes to basal GH release, while the lipoxygenase route predominates in GRF stimulated GH release in vitro. Interestingly, cAMP metabolism can be dissociated from GH release with some of these probes, indicating an action of phospholipid metabolites distal or lateral to the cAMP-generating system. PMID- 2986028 TI - ACTH and corticosterone secretion in rats following removal of the neurointermediate lobe of the pituitary gland. AB - Surgical removal of the neurointermediate lobe of the pituitary gland (NIL-X) in the rat resulted in two abnormalities in ACTH secretion: (1) plasma ACTH and corticosterone levels were elevated in the morning and over a 24-hour period compared with levels in control (NIL-C) rats, and (2) although NIL-X and NIL-C rats had acute increases in plasma ACTH and corticosterone of equal magnitude after interoceptive stimuli (hemorrhage, surgery), NIL-X rats demonstrated markedly smaller elevations in plasma levels of these hormones after neurotropic stimuli (noise, novel environment). This subnormal adrenocortical response of NIL X rats was not due to an impairment in perception of a neurotropic stimulus; these rats had normal latencies to paw licking and to jumping off a heated surface, yet smaller increases in plasma corticosterone after the stimulus. The impairment in ACTH response was not related to stimulus intensity, as NIL-X and NIL-C rats had equal ACTH and corticosterone secretion during both low and high levels of insulin-induced hypoglycemia. NIL-X rats demonstrated a significant elevation in daily water intake, although hematocrit, plasma Na+, K+, osmolality and protein were normal. Significant diurnal rhythms in plasma corticosterone levels and in water intake were maintained as well. The elevated morning plasma ACTH levels, the blunted hormone increases after noise, and the increase in water intake persisted in NIL-X rats 2 months after surgery. These data indicate that removal of the NIL results in (1) chronic elevations in basal ACTH and corticosterone secretion, and (2) chronic impairment in adrenocortical responses to neurotropic stimuli, but not to interoceptive stimuli. The deficit is not due to impaired perceptual capacity nor to the intensity of the stimulus. PMID- 2986029 TI - In vivo evidence for the selectivity of ICI 154129 for the delta-opioid receptor. AB - The in vivo selectivity of the novel delta opioid-receptor antagonist N,N bisallyl-Tyr-Gly-Gly-psi-(CH2S)-Phe-Leu-OH (ICI 154129) was examined in several opioid-selective models. Antagonism at the delta receptor was demonstrated in the striatal head-turn model in the rat. Intrapallidal injection of the relatively selective delta-receptor agonist D-Ala2,D-Leu5-enkephalin (0.5 micrograms) slowed the head-turn time and this effect was completely prevented by prior subcutaneous administration of ICI 154129 (30 mg/kg). The role of delta receptors in two classical test situations was studied using the mixed opioid agonist etorphine and the antagonists naloxone and ICI 154129. The drug ICI 154129 (30 mg/kg, s.c.) failed to prevent the antinociceptive effects and stimulation of locomotor activity produced by etorphine, whereas the relatively selective mu-opioid receptor antagonist, naloxone was effective in both test situations. The possible involvement of delta receptors in morphine-induced dependence was studied by monitoring the abstinence behaviour precipitated in rats given pellets of morphine by either ICI 154129 or naloxone. Naloxone (0.5 mg/kg, i.p.) precipitated a characteristic withdrawal syndrome in conscious rats and, at a much smaller dose (0.02 mg/kg, i.p.), induced shaking behaviour in pentobarbitone anaesthetised rats. No withdrawal signs were observed in either model after injection of ICI 154129 (30 mg/kg, s.c.), suggesting that the delta receptors are not involved in dependence on morphine. PMID- 2986030 TI - Binding characteristics of kappa opioids in rat brain. A comparison of in vitro binding paradigms. AB - Putative kappa binding was investigated in homogenates of the brain of the rat using [3H]ethylketocylazocine and [3H]diprenorphine under conditions where mu and delta sites were blocked. Under blocked conditions, the binding of [3H]ethylketocyclazocine labelled a single site, as defined by kinetic and equilibrium analysis, which showed high affinity for dynorphin 1-17, U50,488H, and benzomorphan compounds. However, blocked binding of [3H]diprenorphine showed a biphasic dissociation curve, and did not show high affinity for the specific kappa agonists dynorphin 1-17 or U50,488H. It is proposed that blocked [3H]ethylketocyclazocine is the more appropriate paradigm to study putative kappa binding, while blocked [3H]diprenorphine may label additional non-mu/non-delta sites. PMID- 2986031 TI - Inhibitory effect of cholesteryl gamma-aminobutyrate on evoked activity in rat hippocampal slices. AB - Cholesteryl gamma-aminobutyrate (C-G) readily crosses the blood-brain barrier and has properties that suggest that it may be a potential gamma-aminobutyric acid (GABA)-mimetic compound. The effect of this compound on the orthodromically evoked discharge of hippocampal pyramidal cells was investigated using slices of rat hippocampus maintained in vitro. The compound produced dose-dependent inhibition of the discharge of pyramidal cells. The magnitude of the inhibitory effect was somewhat less than that produced by a similar dose of GABA, but the duration of the inhibition was prolonged by about 10-fold over that produced by GABA. The inhibition produced by cholesteryl gamma-aminobutyrate was blocked by the addition of picrotoxin to the incubation medium, and by replacement of chloride with isethionate. In addition, pretreatment of slices with the irreversible esterase inhibitor, phenylmethylsulfonylfluoride, attenuated the effects of cholesteryl gamma-aminobutyrate, but not that of GABA. These results suggest that cholesteryl gamma-aminobutyrate has GABA-like actions in the CNS, and that its activity is largely dependent upon enzymatic release of GABA from the compound by esterases present in the tissue. PMID- 2986032 TI - The influence of bicarbonate ions on the GABA-mimetic activity of ethylenediamine. AB - A study was performed to investigate the GABA-mimetic activity of ethylenediamine (EDA) and piperazine at mammalian gamma-aminobutyric acid (GABA) receptors using radioligand binding assays and in vitro isolated tissues. The potency of ethylenediamine and piperazine as inhibitors of the binding of GABA receptors to synaptic membranes from rat brain was measured in Tris-buffers and Krebs Henseleit solution (KHS). The potency of ethylenediamine and piperazine at GABAA and ethylenediamine at GABAB receptors was raised if Krebs-Henseleit solution was used for the assay. Piperazine was inactive at GABAB receptors. The potency of the antagonist of GABAA receptors bicuculline methobromide, was also increased in Krebs-Henseleit when compared with Tris-citrate buffer. Of the ions present in Krebs-Henseleit, bicarbonate ions were responsible for the increase in the GABA mimetic potency of ethylenediamine and piperazine. Addition of either NaHCO3 or KHCO3 (25 mM) to Tris-HCl buffer (for GABAA binding) or Tris-HCl plus 2.5 mM CaCl2 (for GABAB binding) yielded IC50 values similar to those measured in Krebs Henseleit solution. Bicarbonate ions also enhanced the ability of ethylenediamine to potentiate the binding of [3H]diazepam to membranes from rat brain (raising both the potency of ethylenediamine and its maximum effect) in this system. In the absence of HCO-3 ions, ethylenediamine potentiated the binding of [3H]diazepam by raising the maximum binding capacity (Bmax) without changing the affinity (Kd) of the receptors. Potassium bicarbonate (25 mM) caused ethylenediamine to further potentiate the binding of [3H]diazepam by changing both Bmax and Kd.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986033 TI - Treatment of rats with the TRH analog MK-771. Down-regulation of TRH receptors and behavioral tolerance. AB - The regulation of receptors for thyrotropin-releasing hormone (TRH) in the central nervous system (CNS) was studied by administering the TRH analog, MK-771 to rats by three different schedules and then measuring changes in the binding of [3H](3MeHis2) TRH and behavioral responses to a challenge with MK-771. The behavioral responses monitored were wet-dog shakes, large motor movements, small motor movements and forepaw tremor. Temperature changes were also monitored. The first schedule consisted of intracerebroventricular (i.c.v.) administration of MK 771 for seven days (5 micrograms/microliter per hr) via a mini-osmotic pump. At the end of the treatment, rats showed no shaking or large motor movements typically induced by TRH, in response to a 5 mg/kg (i.p.) challenge of MK-771. Receptors were found to be 50% of control levels in the three areas of brain examined. The second schedule consisted of the administration of MK-771 (5 micrograms/2 microliters, i.c.v., once a day and 2 mg/kg, i.p., once a day). It was found that the number of receptors decreased on about the same time course as development of tolerance to wet-dog shakes and large motor movements. The third schedule consisted of the administration of MK-771 (5 micrograms/2 microliters, i.c.v.) once every 2 hr to a total of four doses. These animals eventually developed tolerance to the wet-dog shakes produced by the subsequent challenge with MK-771 and also showed a 50% decrease in receptor binding after the fourth exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986034 TI - Evidence that 2-phenylpyrazolo[4,3-c]-quinolin-3(5H)-one antagonises pharmacological, electrophysiological and biochemical effects of diazepam in rats. AB - The effects of the acute administration of 2-phenylpyrazolo[4,3-c]quinolin-3(5H) one on diazepam-induced behaviour and electrophysiological activity were studied in rat. The compound, in doses of 5-10 mg/kg (i.p.), which per se did not induce alterations in spontaneous locomotor activity, antagonised the sedative effect induced by 5-10 mg/kg (i.p.) of diazepam. The injection of diazepam in rats, induced a profound reduction in the first negative wave of the recording of the visual evoked potential used as a sensitive electrophysiological test, in vivo. 2 Phenylpyrazolo[4,3-c]quinolin-3(5H)-one (10 mg/kg, i.p.) caused a recovery of the amplitude of the first negative wave within a few minutes. This result was confirmed by the finding that 2-phenylpyrazolo[4,3-c]quinolin-3(5H)-one, injected acutely in rats, pretreated with diazepam exhibited the capacity to antagonise the binding of [3H]diazepam determined in vitro on synaptic membrane preparations from cortex. The comparison of the pattern of the visual-evoked potential, recorded after the injection of 2-phenylpyrazolo[4,3-c]quinolin-3(5H)-one (50 mg/kg) with the patterns recorded after the injection of ethyl-8-fluoro-5,6 dihydro-5-methyl-6-oxo-4H-imidazolo(1,5a) (1,4)benzodiazepine-3-carboxylate (50 mg/kg) and ethyl-beta-carboline-3-carboxylate and 1-methyl-beta-carboline demonstrated that 2-phenylpyrazolo[4,3-c]quinolin-3(5H)-one is devoid of intrinsic activity. PMID- 2986036 TI - [3H](-)Baclofen: an improved ligand for GABAB sites. AB - [3H](-)Baclofen, the radiolabelled form of the active isomer of baclofen, has been used as a ligand for GABAB (gamma-aminobutyric acid) receptors on synaptic membranes from whole brain of rat. The pharmacological profile for displacement of this ligand was observed to be identical with that for the racemic ligand [3H](+/-)baclofen and [3H]GABA under conditions where GABAB, but not GABAA sites, were labelled. The displaceable (specific) portion of membrane-bound [3H]( )baclofen was 47.5 +/- 2.3% of the total which was twice that obtained previously with [3H](+/-)baclofen. Two binding components were observed with affinities of 19 and 304 nM and binding capacities of 0.37 and 1.58 pmol/mg protein respectively. It is suggested that [3H](-)baclofen is an improvement over the labelled racemic form and binds to the same sites. It should provide a more reliable tool for studying GABAB receptors. PMID- 2986035 TI - The use of quaternary narcotic antagonists in opiate research. AB - Quaternary ammonium derivatives of narcotic antagonists are commonly used in determining sites of action of opiates in the central nervous system and the periphery because it is widely assumed that they do not readily cross the blood brain barrier, in contrast to their relatively non-polar tertiary counterparts. However, these compounds possess several unique pharmacological properties which have not been taken into consideration in the design of numerous investigations. This article reviews the current state of knowledge concerning the pharmacology of the quaternary narcotic antagonists, examines their use in physiological and behavioral studies of action of opiates, and proposes guidelines for the design of experiments involving these compounds. PMID- 2986037 TI - A comparison of the antinociceptive responses to the GABA-receptor agonists THIP and baclofen. AB - The antinociceptive action of the gamma-aminobutyric acid (GABA) agonists THIP and baclofen was evaluated in mice using hot-plate (48 and 55 degrees C) and tail immersion (50 degrees C) procedures. It was found that atropine reversed antinociception induced by THIP but not that induced by baclofen in the 48 degrees C test, whereas the anticholinergic drug blocked the response to both GABA agonists when the stimulus was provided by a 55 degrees C hot-plate. Atropine methylnitrate, mecamylamine, picrotoxin and bicuculline had no effect on antinociception induced by THIP or baclofen. Prior treatment with haloperidol enhanced only the response to baclofen on the 55 degrees C hot-plate. A reciprocal cross-tolerance was found between THIP and baclofen in the tail immersion assay, although only THIP exhibited cross-tolerance to morphine. These results suggest that while the analgesic response to THIP and baclofen is partially mediated by a common system, the two agents act by independent mechanisms as well. PMID- 2986039 TI - Pharmacology of the corticocaudate excitatory postsynaptic potential in the cat: evidence for its mediation by quisqualate- or kainate-receptors. AB - Caudate neurons were recorded with intracellular electrodes in halothane anaesthetized cats during microiontophoretic application of drugs and simultaneous stimulation of the corticocaudate pathway. Application of 2-amino-7 phosphonoheptanoic acid inhibited excitations induced by the N-methyl-D-aspartic acid receptor agonists N-methyl-D-aspartic acid and quinolinic acid, but not those elicited by quisqualic acid or activation of the cortico-caudate pathway. Selective inhibition of N-methyl-D-aspartic acid induced excitations was also found in vitro in the frog hemisected spinal cord preparation where a pA2-value of 5.5 against N-methyl-D-aspartic acid was determined. The endogenous tryptophan metabolite, kynurenic acid, antagonized excitations induced by N-methyl-D aspartic, quisqualic, L-glutamic and kainic acid as well as the excitatory postsynaptic potential (EPSP) evoked in caudate cells by stimulation of the corticocaudate pathway, while action potentials elicited by an intracellularly applied depolarizing current were only slightly affected. In vitro experiments with the frog hemisected spinal cord preparation suggested that kynurenic acid might be a competitive antagonist of both N-methyl-D-aspartate and quisqualate receptors, with pA2-values of about 4.8 and 4.0, respectively. From these results it is concluded that the three-receptor concept for excitatory amino acids are proposed by Watkins and colleagues is probably applicable to the cat caudate nucleus and that the cortically evoked monosynaptic EPSP is mediated by a non-N methyl-D-aspartate quisqualate- or kainate-receptor. PMID- 2986038 TI - Parallel changes in the sensitivity of gamma-aminobutyric acid and noradrenergic receptors following chronic administration of antidepressant and GABAergic drugs. A possible role in affective disorders. AB - Chronic treatment with the antidepressants imipramine or nomifensine, or the gamma-aminobutyric acid (GABAergic) agents, baclofen or THIP, produced a decrease in the Bmax for binding sites of GABAergic and noradrenergic receptors. Chronic treatment with imipramine or nomifensine produced a decrease in the Bmax of both binding of [3H]dihydroalprenolol and [3H]GABA receptors in the cerebral cortex and hippocampus. Chronic treatment with baclofen or THIP also produced a decrease in the Bmax of binding of [3H]dihydroalprenolol receptor in the cerebral cortex and hippocampus. These results suggest a possible link between the GABAergic and noradrenergic systems, which may be important in understanding the mechanism of action of antidepressant drugs, and suggests a possible role for GABA in affective disorders. PMID- 2986041 TI - Effects of topical baclofen on C fibre-evoked neuronal activity in the rat dorsal horn. AB - Baclofen appears to be an agonist for the bicuculline-insensitive gamma aminobutyrateB receptors associated with C fibre terminals in the dorsal horn of the spinal cord. We have tested the effect of baclofen (applied intrathecally onto the spinal cord) on the A and C fibre-evoked responses of convergent/multireceptive neurones in the halothane-anaesthetized rat. L-Baclofen produced a dose-dependent inhibition of the C fibre- and pinch-evoked activity of these neurones which persisted for 2 h whilst the A fibre and tactile activities were little changed. The C fibre-evoked (X 3 threshold) responses were markedly or completely inhibited 10 min after doses of between 0.25 and 30 micrograms of L baclofen (n = 21) with 0.05 micrograms causing a 48% (n = 3) and 0.01 micrograms a 28% inhibition (n = 3). D-Baclofen (30 micrograms), the inactive isomer, produced no significant changes in activity (n = 10). Bicuculline (60 micrograms) applied intrathecally before (n = 7) or after (n = 8) L-baclofen did not reverse the inhibitions. Intravenous baclofen (1-3 mg/kg) also produced neuronal inhibitions similar to the effects of intrathecal injection. The results suggest that gamma-aminobutyrateB receptors may exert a presynaptic control of C fibre afferents in the dorsal horn following intrathecal administration in the rat. PMID- 2986040 TI - Stroboscopic illumination and dark rearing block the sharpening of the regenerated retinotectal map in goldfish. AB - Blocking activity with intraocular tetrodotoxin prevents the sharpening of the retinotectal map formed during regeneration of the optic nerve. If (under normal conditions) the initially diffuse map sharpens because of correlated activity in neighboring but not distant ganglion cells, then sharpening should also be prevented merely by disrupting the spatiotemporal correlation in the pattern of activity. To test this idea, fish were exposed during regeneration to stroboscopic illumination in a featureless environment, or were maintained in complete darkness. The regenerating cells remained visually responsive after axotomy, and the xenon strobe effectively drove each ganglion cell at a constant latency. The maps formed in the strobe-reared fish were normally oriented, but the multiunit receptive fields were greatly enlarged, averaging 32 degrees. In control regenerates, multiunit receptive fields averaged only 11-12 degrees, nearly the same as for single units. Dark rearing, which allows only spontaneous activity, also resulted in enlarged multiunit receptive fields, averaging more than 28 degrees. Both effects parallel those reported previously with tetrodotoxin block. The mature projection did not become diffuse as a result of the strobe rearing, and the sensitive period corresponded to the early stage of synaptogenesis (20-34 days). Periods of normal visual exposure after 35 days produced very little sharpening of the diffuse maps produced during either strobe or dark rearing. The results are attributed to an activity-dependent stabilization of developing synapses. The correlated firing of neighboring ganglion cells could allow postsynaptic summation of their responses, and the retention of those more effective, retinotopically placed synapses might then occur via a Hebbian mechanism. PMID- 2986042 TI - The ionic mechanism of postsynaptic inhibition in motoneurones of the frog spinal cord. AB - The ionic mechanism of postsynaptic inhibition in frog spinal motoneurones was studied with conventional and with ion-sensitive microelectrodes. In these neurones the inhibitory postsynaptic potential was depolarizing, its reversal potential being 15 mV less negative than the resting membrane potential. During the inhibitory postsynaptic potential the input resistance of the motoneurones was reduced to 20% of the resting value, indicating a strong increase of membrane conductance. The Cl- equilibrium potential calculated from intra- and extracellular Cl- activity measurements coincided with the reversal potential of the inhibitory postsynaptic potential to within a few millivolts. During repetitive inhibitory postsynaptic activity the intracellular Cl- activity decreased markedly, while the extracellular Cl- activity increased slightly. These changes of intra- and extracellular Cl- activities were no longer found after suppression of the inhibitory postsynaptic potential by strychnine. Blockade of an active, inward-going Cl- transport system in motoneurones by NH+4 led to a shift of the Cl- equilibrium potential and the reversal potential of the inhibitory postsynaptic potential towards the resting membrane potential. After prolonged action of NH+4, the Cl- equilibrium potential approached the membrane potential to within 5 mV, while the reversal potential of the inhibitory postsynaptic potential and resting membrane potential coincided. The difference between Cl- equilibrium potential and membrane potential after blockade of the Cl pump is traced back to interfering intracellular ions, such as HCO-3 or SO42-, leading to an overestimation of intracellular Cl- activity and to the calculation of an erroneous Cl- equilibrium potential. Inhibitory amino acids like gamma aminobutyrate or beta-alanine evoked depolarizations with reversal potentials similar to that of the inhibitory postsynaptic potential. These depolarizations were associated with a marked decrease of neuronal input resistance during inhibition. During the actions of these compounds a decrease of intracellular and a small increase of extracellular Cl- activity were found. The activities of other ions (K+, Ca2+ and Na+) did not change significantly, with the exception of extracellular K+ activity, which was slightly increased. Evidence is presented that the inhibitory postsynaptic potential, as well as the depolarizing action of inhibitory amino acids in motoneurones, is the result of an increase in membrane Cl- permeability and an efflux of Cl- from these cells, while other ions do not seem to be involved. PMID- 2986043 TI - Dual effects of the snake venom polypeptide vipoxin on receptors for acetylcholine and biogenic amines in Aplysia neurons. AB - Vipoxin, a 13,000-dalton polypeptide component of Russell's viper venom, has a dual pattern of effects on the responses of voltage-clamped Aplysia neurons to acetylcholine and biogenic amines. Application of low doses of vipoxin by pressure ejection reversibly antagonized all three types of ionic response to acetylcholine and carbachol. The blockade by vipoxin of acetylcholine responses was not prevented by eserine. The order of susceptibility of acetylcholine responses to blockade by vipoxin was Na+ greater than K+ greater than Cl-. Low doses of vipoxin also produced a reversible potentiation of excitatory responses to dopamine with a slower time course of onset and recovery. Inhibitory responses to dopamine (Cl-, K+) and both excitatory and inhibitory responses to histamine and 5-hydroxytryptamine were little affected by vipoxin. Higher doses of vipoxin directly evoked current responses which were always of the same ionic type as that evoked by acetylcholine or carbachol. Responses to cholinergic agonists and vipoxin were both blocked by cholinergic antagonists but not by antagonists of biogenic amine receptors, which reversibly antagonized the responses to amines on the same cell. These results suggest that vipoxin, which has no demonstrated actions on vertebrate acetylcholine receptors, acts as a partial agonist at all three types of acetylcholine receptor in Aplysia neurons. Our observations thus provide evidence for some degree of phylogenetic difference between vertebrate and molluscan acetylcholine receptors. PMID- 2986044 TI - Regulation of dopamine release by impulse flow and by autoreceptors as studied by in vivo voltammetry in the rat striatum. AB - Extracellular dopamine concentration has been monitored in the striatum of pargyline treated, anaesthetized rats using differential normal pulse voltammetry. The catechol oxidation current recorded with electrochemically treated carbon fiber electrodes disappeared when the dopaminergic terminals were selectively destroyed by 6-hydroxydopamine. Calibration of the basal oxidation current revealed that the extracellular dopamine concentration was 26 nM. Brief and moderate electrical stimulation of the nigrostriatal pathway at the level of the medial forebrain bundle induced a large increase in the dopamine current. The observed elevation in the dopamine signal lasted as long as the stimulation. It varied with the frequency (0-25 Hz) of the pulses in an exponential manner. Stimulation pulses distributed in a bursted pattern were twice as potent as an equivalent number of pulses regularly spaced. High frequency stimulations (50 Hz) were also investigated in anaesthetized rats (without pargyline) with untreated carbon fiber electrodes; they induced a very large increase in the dopamine extracellular concentration (up to 8-15 microM). Interruption of the dopaminergic impulse flow either by an electrolytic lesion or by a low dose of apomorphine (0.05 mg/kg) caused an immediate decrease of the dopamine current. The time courses and amplitudes (-70%) of these effects were identical. Subsequent injection of haloperidol (0.5 mg/kg) reversed the apomorphine effect up to +360% of the control basal value. Administration of dopaminergic antagonists such as haloperidol (0.05 and 0.5 mg/kg) or metoclopramide (2 mg/kg) significantly increased the dopamine current up to 317, 340 and 215% of the respective control values. Nomifensine (4 mg/kg) produced a big increase (+417%) of the extracellular dopamine levels. The effect of electrical stimulation of the dopaminergic pathway was potentiated by drugs such as amphetamine (2 mg/kg), nomifensine (4 mg/kg) or haloperidol (0.05 and 0.5 mg/kg) but was not altered by apomorphine (0.05 mg/kg). The study by in vivo voltammetry of the variations in the striatal extracellular dopamine concentrations shows that the release of dopamine is under the influence of both the frequency of impulse flow and of dopaminergic striatal autoreceptors. PMID- 2986045 TI - The morphogenesis of stereotyped behavior induced by the dopamine receptor agonist apomorphine in the laboratory rat. AB - The seemingly unrelated stereotyped locomotor "acts" reported in the literature to be produced by apomorphine in rats are shown to be composites, whose form and sequence are determined by the particular values of a few component variables which form a common denominator in each of the behaviors. Three variables, continuous snout contact, forward progression and turning, account for much of the behavior. In the course of the drug's action these emerge in succession and vary in amount, the latter two successively reaching a peak and subsiding. The interaction between forward progression and turning yields in sequence, forward walking, circling, revolving, tight pivoting and finally side-to-side movements of the forequarters around the relatively stationary hindquarters. Later behaviors in this list are gradually incorporated into the sequence as earlier ones are eliminated. The course of change in forward progression and turning is also reflected in changes in the sequence and in the direction of stepping of each of the four legs. The order in which the behavior unfolds under the drug is opposite to that manifested in ontogeny and in recovery from lateral hypothalamic damage, suggesting that at the particular high dose used, apomorphine is acting not only to activate the behavior but also to shut it down. PMID- 2986046 TI - Peripheral neuropathy, high serum IgM, and paraproteinemia in mother and son. AB - A mother and son had peripheral neuropathy, abnormal elevation of serum IgM, and paraproteinemia. These patients may have a familial disorder of immune regulation with antibody-mediated neuropathy or a familial neuropathy with a secondary immune response. PMID- 2986047 TI - Infantile neuroaxonal dystrophy: perinatal onset with symptoms of diencephalic syndrome. AB - In a neonatal case of infantile neuroaxonal dystrophy, there was emaciation, nystagmus, and endocrinologic disorder suggesting the diencephalic syndrome. At autopsy, spheroid bodies were widely disseminated, particularly in the hypothalamus, infundibulum, and neurohypophysis. The pathologic process may have started in utero. PMID- 2986049 TI - [Loco-regional recurrence of cancer of the rectum]. PMID- 2986048 TI - Dementia without Alzheimer pathology. AB - We studied five demented patients who, on neuropathologic examination, had cell loss and Lewy bodies in substantia nigra and locus ceruleus and few Alzheimer type changes. The nucleus basalis had minimal cell loss in three patients and was not available in two. The lesions in the substantia nigra and locus ceruleus were unlikely to account for the dementia, and other structural or biochemical derangements, probably cortical but possibly subcortical, must also have been present but not visible at the light microscopic level. PMID- 2986050 TI - [Treatment of cancer of the rectum]. PMID- 2986051 TI - [Pulmonary surgery today. Considerations on reconstructive bronchial surgery]. PMID- 2986052 TI - [Idiopathic hemochromatosis]. AB - Idiopathic hemochromatosis is a hereditary disease characterized by a progressive iron overload secondary to high intestinal iron absorption. After a latent period of many years, manifestations of liver cirrhosis, diabetes mellitus, cardiac failure, hypogonadism, skin hyperpigmentation and arthropathy can occur. Liver cirrhosis is the most common feature and it is complicated by hepatocellular carcinoma in 30% of cases. Tests of high sensibility are available for early diagnosis. Repeated phlebotomy can prevent clinical features in asymptomatic patients and can improve prognosis in symptomatic subjects. Current concepts in idiopathic hemochromatosis are reported in this review. PMID- 2986053 TI - A transient increase in the activity of CA3 neurons induces a long-lasting reduction in the excitability of Schaffer collateral terminals in rat hippocampus. AB - Following high frequency discharges of CA3 neurons, a long-lasting potentiation (LLP) of the CA1 neuronal population spike results. Subsequent to a transient increase in the firing rate of CA3 neurons produced by the application of DL homocysteate on the soma in the presence of normal medium but not in the presence of Ca2+ -free (Mn2+ -containing) medium, the threshold for antidromic activation in the CA1 area of the neuron was increased for a prolonged period of time. Since the induction of the presynaptic terminal excitability reduction as well as LLP of the CA1 population spike are Ca2+ -dependent, a cause-and-effect relationship may exist between these two processes. PMID- 2986054 TI - Effect of DN-1417, a synthetic thyrotropin-releasing hormone analogue, on [3H]GABA binding in the cerebellum of ataxic rats. AB - The neonatal administration of cytocine arabinoside (40 mg/kg, s.c.) induced marked ataxia and hypoplastic cerebellum in male Sprague-Dawley rats. Specific [3H]GABA binding in the cerebellum was significantly reduced by cytosine arabinoside, whereas [3H]GABA binding in the cerebral cortex was not changed. The administration of DN-1417 (1 mg/100 g body wt., i.p.), a synthetic derivative of thyrotropin-releasing hormone (TRH), significantly increased [3H]GABA binding in the cerebellum of cytosine arabinoside-induced ataxic rats. These results suggest that TRH may play a role in regulating GABA receptors involved in the function of the rat cerebellum. PMID- 2986055 TI - Effects of fasciculin 2, an anticholinesterase polypeptide from green mamba venom, on neuromuscular transmission in mouse diaphragm preparations. AB - Fasciculin 2, a polypeptide from green mamba (Dendroaspis angusticeps) venom, causes an increase in the twitch response of mouse phrenic nerve-hemidiaphragm preparations to indirect stimulation. Intracellular recording reveals that fasciculin 2 augments neuromuscular transmission by increasing the amplitude and duration of endplate potentials. Its action is not reversed by washing. Interactions with neostigmine confirm that fasciculin 2 acts as an anticholinesterase. It has no presynaptic actions on transmitter release or postsynaptic receptor blocking actions. On chicken muscle preparations, fasciculin 2 has no anticholinesterase actions. Because of this selectivity and its apparent irreversibility, fasciculin 2 should be useful in characterizing different forms of acetylcholinesterase. PMID- 2986056 TI - Pro-opiomelanocortin peptides in the human hypothalamus: comparative study between normal subjects and Parkinson patients. AB - The concentrations of gamma 3-melanotropin-, alpha-melanotropin-, corticotropin-, gamma-lipotropin- and beta-endorphin-immunoreactivities were determined simultaneously, before and after gel exclusion chromatography, in whole hypothalamic extracts of normal subjects and Parkinson patients. All five immunoreactivities were present and were all significantly correlated to each other. Shorter peptides (alpha-melanotropin, gamma 3-melanotropin, beta-endorphin and a lipotropin37-58-like peptide) were the dominant products. Whereas the dopamine content was significantly reduced in Parkinson patients, there was no significant difference for any peptide between normal subjects and Parkinson patients, either in tissue concentrations or in chromatographic patterns. PMID- 2986057 TI - The importance of the striato-nigro-collicular pathway in the expression of haloperidol-induced tonic electromyographic activity. AB - Experiments were carried out to investigate the participation of the striatonigral and nigrocollicular GABAergic pathways in the expression of haloperidol-induced tonic electromyographic (EMG) activity of the gastrocnemius soleus muscle in awake rats. It was shown that bilateral injections of haloperidol (500 ng/0.5 microliter) into the rostral part of the neostriatum induced a tonic EMG activity which could be significantly reduced by a second bilateral injection of muscimol (25 ng/0.5 microliter) into the substantia nigra pars reticulata (SNR) or of bicuculline (50 ng/0.5 microliter) into the intermediate or deep layers of the colliculus superior (CS). These results suggest that the striatonigral and nigrocollicular GABAergic pathways play a critical role in the transmission of haloperidol-induced tonic EMG activity. PMID- 2986058 TI - Products of cholecystokinin (CCK)-octapeptide proteolysis interact with central CCK receptors. AB - Peptidases present in central nervous system (CNS) synaptic membranes, hydrolyze the neuroactive peptide cholecystokinin-octapeptide (CCK-8; Asp-Tyr-SO3H-Met-Gly Trp-Met-Asp-Phe-NH2). In order to determine the pathway of degradation, synthetic CCK-8 was incubated at 37 degrees C with purified synaptic membranes; at various intervals reaction samples were removed from the reaction mixture and analysed by high-performance liquid chromatography to identify and quantify the peptide fragments. The results indicate an initial endopeptidase cleavage at the Met-Gly bond producing CCK-5 (Gly-Trp-Met-Asp-Phe-NH2). The carboxyl-terminal pentapeptide is further proteolysed to CCK-4 (Trp-Met-Asp-Phe-NH2) by a puromycin sensitive aminopeptidase and to CCK-3 (Met-Asp-Phe-NH2) and Gly-Trp by an endopeptidase action. CCK-3 and CCK-2 appear to be relatively stable end products. Moreover, these proteolytic fragments are shown to bind to the CCK receptor in brain with varying potencies. PMID- 2986059 TI - Muscle relaxant action of 2-chloroadenosine in genetically spastic rats is independent of gamma-aminobutyric acid-mediated inhibition. AB - The role played by benzodiazepine (BDZ) receptors and GABAergic mechanisms in the muscle relaxant effect of the adenosine receptor agonist, 2-chloroadenosine (2 CLA) was evaluated in genetically spastic rats. 2-CLA reduced in a dose-related manner the spontaneous tonic activity in the electromyogram (EMG) of the gastrocnemius-soleus muscle. While the muscle relaxant effect produced by the adenosine analogue was abolished by aminophylline, a methylxanthine with potent antagonistic activity of adenosine-mediated inhibition, the two gamma aminobutyric acid (GABA) antagonists bicuculline and picrotoxin did not reverse it. Neither Ro 15-1788 nor CGS 8216, specific BDZ receptor antagonists, nor beta carboline-3-carboxylic acid methyl ester, an inverse agonist at BDZ receptors, affected the depressant effect of 2-CLA in the EMG. These results suggest that 2 CLA does not interact with GABA or BDZ receptor-mediated events in vivo to produce the muscle relaxant action. It is proposed that purinergic mechanisms may be involved in the muscle relaxant effects of a variety of drugs. PMID- 2986060 TI - Morphological variability, lectin binding and Na+,K+-activated adenosine triphosphatase activity of isolated Muller (glial) cells from the rabbit retina. AB - Rabbit retinal Muller cells were isolated by means of papaine and mechanical dissociation. These cells were shown to have a well preserved morphology and to preserve viability for many hours. Intense wheat germ agglutinin binding occurs on the photoreceptor side of Muller cells, especially in the microvillous region. Rabbit retinal Muller cells have a Na+,K+-activated adenosine triphosphatase activity in the same order of magnitude as brain astroglial cells. PMID- 2986061 TI - Are the anxiogenic effects of yohimbine mediated by its action at benzodiazepine receptors? AB - Yohimbine, which has activity both at alpha 2-adrenoreceptors and benzodiazepine receptors in the CNS, had an anxiogenic action (1.25-2.5 mg/kg) in the social interaction test in rats. This effect was reversed by the alpha 2-adrenoceptor agonist clonidine (0.01 mg/kg), but not by the benzodiazepine receptor agonist chlordiazepoxide (5-10 mg/kg) or the antagonist Ro 15-1788 (10 mg/kg). These results suggest that the anxiogenic effects of yohimbine are not attributable to its low affinity effects at benzodiazepine receptors but to its alpha 2 adrenoceptor antagonist activity. PMID- 2986062 TI - Effect of acute acrylonitrile exposure on metrazol induced seizures in the rat. AB - The effects of acute exposure to acrylonitrile (ACN), 10, 20, or 40 mg/kg by gavage, on the ability of metrazol (MTZ) to induce seizures was studied in adult, male Sprague-Dawley rats. The frequency of seizure occurrence and the frequency of a lethal seizure was greater when the high ACN dosage was given in combination with metrazol. This dosage of ACN was not lethal when given alone. Examination of brain tissue in these animals revealed no difference in cyanide levels when MTZ was combined with ACN. However, brain cytochrome c was significantly lower in animals given ACN+MTZ and brain cholinesterase was significantly higher. These results suggest that the enhanced lethality occurring in animals exposed to the combination of ACN+MTZ is not due to cyanide, a metabolic product of ACN, but rather to a potentiation of other effects of ACN perhaps involving cholinergic neurotransmission. PMID- 2986063 TI - Transferrin and its cellular receptor underscore the importance of iron in cell growth. PMID- 2986065 TI - Acute HTLV III infection. PMID- 2986064 TI - Cis-platinum and distal renal tubule toxicity. AB - Glomerular filtration rate (GFR) and plasma electrolyte concentrations were evaluated prospectively in 22 consecutive patients treated with serial courses of cisplatinum combination chemotherapy. In addition, renal distal tubular function was assessed in the first 12 by measurement of overnight urinary concentration after intranasal desamino-d-arginine vasopressin inhalation and by a standard short urinary acidification test following the administration of oral ammonium chloride. Cumulative and dose-related hypomagnesaemia was observed in all patients but was unassociated with clinical symptoms. No significant fall in GFR was seen in the group as a whole but four of the 12 patients studied more comprehensively developed impairment of urinary concentrating and acidifying abilities, independent of any change in GFR. This indicates a selective effect on the renal distal tubule. PMID- 2986066 TI - AIDS and organ transplantation. PMID- 2986067 TI - Alternative health care. Selected individual therapies. PMID- 2986068 TI - Acute leukemia secondary to lung cancer. Case report and review of the literature. AB - A case of acute myelomonocytic leukemia following cytotoxic therapy for an oat cell carcinoma of the lung in a 48-year-old man is reported. This case is characterized by a long phase of increasing macrocytosis during cyclophosphamide maintenance therapy. 15 other cases of secondary leukemia to lung cancer from the literature are reviewed. All patients received alkylating agents. Most patients showed peripheral blood changes more than 3 months before the onset of acute leukemia. All leukemia cases, except one, were nonlymphocytic with a high frequency of erythroleukemia. The development of acute leukemia seems to be linked with an unexpected good response to chemotherapy in advanced or poor histologic prognosis cancers. With therapeutic improvement in lung cancer, secondary leukemia could become a major hazard as in other cancers successfully treated with cytotoxic agents. PMID- 2986069 TI - Ocular and orbital toxicity following intracarotid injection of BCNU (carmustine) and cisplatinum for malignant gliomas. AB - Eleven patients treated with intracarotid BCNU, cisplatinum, or BCNU and cisplatinum in combination for recurrent malignant gliomas were followed with serial ophthalmologic examinations for 2 to 11 months. Eight patients developed significant visual loss ipsilateral to the side of infusion. Secondary glaucoma and internal ophthalmoplegia were new complications observed after BCNU treatment. An unusual pigmentary retinopathy, previously unreported, was seen in patients treated with cisplatinum. One patient also developed a cavernous sinus syndrome after the intracarotid administration of cisplatinum. PMID- 2986070 TI - [Use of technetium Tc 99m pyrophosphate for the examination of patients with medial femoral neck fracture]. PMID- 2986071 TI - Microneuronography and its relation to perceived sensation. A critical review. PMID- 2986072 TI - Ketamine analgesia is not related to an opiate action in the periaqueductal gray region of the rat brain. AB - Ketamine is an injectable anesthetic agent that has been shown to interact as an agonist at opiate receptors. In addition, its antinociceptive action in rats is antagonized by the narcotic receptor antagonist naloxone. Thus it was assumed that the anesthetic may activate the pain inhibitory pathway, originating in the periaqueductal gray (PAG) and descending into the spinal cord, in a manner similar to that of narcotics like morphine. In the present study, it was verified that the systemic administration of naloxone (3 mg/kg i.p.) antagonized the elevation in tail-flick latency produced by an anesthetic dose of ketamine (160 mg/kg i.p.), but did not alter the duration of anesthesia (defined as duration of the loss of the righting reflex). However, when naloxone (3 micrograms/0.5 microliter/30 sec) was given by microinjection into the PAG it was found to be ineffective against the ketamine-induced elevation of the tail-flick latency. In contrast, the microinjection of the antagonist significantly attenuated (halved) the elevated latency in response to systemically administered morphine (4 mg/kg s.c.). It was also shown that ketamine was unable to elicit an increase in the latency of the tail-flick reflex when administered directly into the PAG over a wide range (0.10-100 micrograms) of doses. On the other hand, a local anesthetic like effect of ketamine, known to occur when the drug is used in high concentration, was observed when doses exceeding 0.1 microgram were injected into the PAG. This action interfered with opiate actions in the PAG and made data from the microinjection studies difficult to interpret. The descending, pain inhibitory neuronal system originating in the PAG does not appear to participate in the antinociceptive action of ketamine measured by the tail-flick reflex. Perhaps the drug's effects are associated with alternative opiate mechanisms and/or opiate receptor subtypes not present on the cells of origin of the descending nerves within the PAG. PMID- 2986073 TI - Acupuncture analgesia. PMID- 2986075 TI - Autopsy findings in the acquired immune deficiency syndrome. PMID- 2986074 TI - The histopathology of the acquired immune deficiency syndrome. PMID- 2986076 TI - Integrated imaging of neonatal renal masses. AB - Thirty-three neonatal renal masses were evaluated during a 2-year interval. The final diagnoses in these 33 patients were hydronephrosis [14], multicystic dysplastic kidney [10], renal vein thrombosis [3], obstructed upper pole duplication [2], polycystic kidney disease [2], nephroblastomatosis [1], and mesoblastic nephroma [1]. We recommend an integrated imaging approach that utilizes sonography to clarify anatomy and renal scintigraphy or excretory urography to determine renal function. PMID- 2986077 TI - Child care workers and children with congenital cytomegalovirus infection. PMID- 2986079 TI - Binding characteristics of dermorphin, and [dermorphin1-7]-beta c-endorphin in rat brain membranes. AB - Dermorphin and a camel beta-endorphin (beta c-EP) analog in which residues 1-7 correspond to the dermorphin sequence ([Dermorphin1-7]-beta c-EP) have been investigated with respect to their receptor binding characteristics using human and camel beta-EP as reference peptides. Tritiated dihydromorphine, [D-Ala2, D Leu5]-enkephalin, ethylketocyclazocine and human beta-endorphin were used as primary ligands in the rat brain membrane preparation for radioreceptor assay. Camel beta-endorphin was the most potent peptide in all experiments. [Dermorphin1 7]-beta c-EP is significantly less potent towards 3H-ethylketocyclazocine and 3H [D-Ala2, D-Leu5]-enkephalin but is as potent towards 3H-dihydromorphine and 3H human beta-endorphin. Dermorphin itself weakly displaces tritiated dihydromorphine, [D-Ala2, D-Leu5]-enkephalin and ethylketocyclazocine (potency relative to camel beta-EP, 1-4%) but it is more potent (9%) in competition with tritiated human beta-endorphin. Dermorphin and the [Dermorphin-1-7]-beta c-EP appear to interact preferentially with mu opiate receptors. PMID- 2986080 TI - The postnatal development of VIP binding sites in rat forebrain and hindbrain. AB - The specific binding of 125I-labeled vasoactive intestinal peptide (VIP) to brain membranes from the forebrain and hindbrain regions of 2 to 37-day postnatal rats was measured. In both regions of the brain, VIP binding was low but detectable two days after birth and rose markedly between postnatal days 7 and 17. This increase in VIP binding with age correlates well with observed increases in VIP stimulated adenylate cyclase activity and increases in VIP content as determined by radioimmunoassay. In hindbrain, the density of VIP binding sites was substantially higher than the forebrain at two days, while at 37 days, they were about equal, suggesting that the hindbrain regions may mature neurochemically prior to the forebrain. Total binding sites for forebrain and hindbrain were about equal at birth for both brain regions, while forebrain had a substantially greater number of sites at 37 days postnatal. The presence of VIP binding sites in both forebrain and hindbrain early in postnatal development suggests that VIP may play a role in development of the brain. PMID- 2986078 TI - Photochemically produced intracellular concentration jumps of cAMP mimic the effects of catecholamines on excitation-contraction coupling in frog atrial fibers. AB - Previously, we reported that concentration jumps of cAMP produced by light flashes in the presence of a photosensitive analogue of cAMP increase the amplitude of the slow inward current (Isi) in isolated bullfrog atrial trabeculae (Nargeot et al. 1983). Here, using newly designed photolabile cyclic nucleotides (Nerbonne et al. 1984a), we have examined the effects of intracellular concentration jumps of cAMP and cGMP on excitation-contraction coupling in frog heart. Concentration jumps of cAMP increase the amplitude and the duration of action potentials, increase Isi and twitch tension. Following single flashes, maximum responses are observed in 10-30 s and recovery times are 30-120 s. The time courses of the cAMP-induced increases in Isi and phasic tension amplitudes are parallel, implying a direct correlation between Ca2+ influx through the slow channels and the development of phasic tension. Although the amplitudes are increased severalfold, cAMP jumps do not measurably alter the kinetics or voltage dependences of the current or tension. cAMP concentration jumps increase the delayed K+ current (IK) and decrease tonic tension; relaxation of contraction is not, however, influenced by cAMP jumps. Concentration jumps of cGMP, on the other hand, have no measurable effects on the action potential, Isi, IK or tension in this preparation. PMID- 2986081 TI - Reduced ACTH, while normal beta-endorphin CSF levels in early epileptic encephalopathies. AB - Since ACTH and the opioids display opposite effects on experimentally-induced seizures, cerebrospinal fluid (CSF) levels of ACTH and beta-endorphin (beta-EP) were measured in 6 children (4-8 months) affected by infantile spasms with hypsarhythmia, an idiopathic early onset encephalopathy, and in 8 age-matched controls. beta-EP levels in the patients (76.3 +/- 14.7 fmol/ml, M +/- SD) did not differ from those in controls (109.8 +/- 42.7) while babies with epileptic encephalopathy showed reduced ACTH levels in the CSF (3.8 +/- 1.5) as compared to controls (9.0 +/- 3.7, p less than 0.01). This resulted in an increased beta EP/ACTH ratio. Another patient previously treated with ACTH showed a normal CSF level of ACTH (9.0) with a normal beta-EP/ACTH ratio while in clinical and EEG remission. These results are consistent with the hypothesis that some infantile seizures unrelated to brain injuries could originate from an ACTH deficiency at central level and/or an imbalance of neuropeptidergic pathways. PMID- 2986082 TI - Vasopressin receptor subtypes in dorsal hindbrain and renal medulla. AB - We have investigated the ability of a series of synthetic vasopressin analogues and related peptides to compete with (3H)-arginine8 vasopressin for binding sites in rat renal medulla and dorsal hindbrain. In renal medulla, arginine8 vasopressin and deamino arginine8 vasopressin, a selective antidiuretic, were equipotent while two antagonists of the pressor action of arginine vasopressin were less potent. In the dorsal hindbrain, arginine8 vasopressin and the pressor antagonists were more potent than the synthetic antidiuretic. Potency profiles of these and other analogues suggest that the renal medulla and dorsal hindbrain vasopressin receptors represent different subtypes. PMID- 2986083 TI - Psychobiology of anxiety and anxiety disorders. AB - New techniques for studying receptor pharmacology, neurotransmitter activity, and neuroendocrine function in affective illness have made it possible to carry out sophisticated neurochemical and neuropharmacologic investigations of the anxiety disorders. Some important reasons for pursuing these strategies have been the high frequency of depression in anxious patients, the effectiveness of antidepressants in the treatment of panic disorder, and the availability of probes for studying the physiologic changes that occur during anxiety states in human subjects. In addition, the ability reliably to induce anxiety states in man has made it possible to study at least some clinical forms of anxiety under laboratory conditions. Although animal models for simple phobia have been developed, there are currently no adequate animal models of panic disorder in man. If valid, reliable animal models, for panic disorder and other human anxiety disorders can also be identified, then a much better understanding of the nature and causes of anxiety and more effective diagnosis and treatment of clinically important anxiety disorders may be possible. PMID- 2986084 TI - Effect of long-term hyperprolactinemia on the prolactin receptor content of the rat ventral prostate. AB - The rat ventral prostate contains prolactin receptors, and during sexual maturation prolactin stimulates the growth of this gland. The aim of the present investigation was to evaluate whether prolactin is involved in the regulation of the number of its own receptor sites in the rat ventral prostate. To this end, the content of prolactin receptors was estimated in prostate membranes of control and chronically hyperprolactinemic rats both before and after in vitro desaturation with 4 M MgCl2. Hyperprolactinemia resulted in a 40% increase in the number of available prolactin receptors (P less than 0.05). In vitro desaturation of receptors resulted in loss of 84% of protein and 36 +/- 6% and 52 +/- 6% of prolactin receptors from ventral prostate membranes of control and hyperprolactinemic rats respectively (P less than 0.05). We have concluded that the rat ventral prostate membranes are not suited to in vitro desaturation of prolactin receptors with MgCl2. From the increase in the number of available prolactin receptors after hyperprolactinemia we have concluded that prolactin is involved in the regulation of the number of its own receptors in the rat ventral prostate. PMID- 2986085 TI - Responses of chickens lacking or expressing endogenous avian leukosis virus genes to infection with exogenous virus. AB - Line 0 chickens that lack endogenous viral (ev) genes and 15I5 X 7 chickens that carry and express ev genes were inoculated with the avian leukosis virus, RAV-1, at 1 day of age or as embryos, and maintained to 70 days of age. Comparable uninoculated controls were also kept. Most of the day-old inoculated chickens developed antibody by 70 days, but as expected, most of the embryo inoculated chickens remained viremic. Major differences in response between the lines were only noted after day-old inoculation. About 40% of Line 0 died of a specific nonneoplastic syndrome (NNS) while none of the 15I5 X 7 chickens did. Line 0 also developed neutralizing antibodies at a much earlier age than 15I5 X 7 and, consequently, virus was cleared from the serum much earlier. Day-old RAV-1 infection of Line 0 also lowered hematocrit more at 14 and 21 days of age than in 15I5 X 7 chickens. Body weight was clearly reduced at all ages in both lines of chickens by both day-old and embryo inoculation. After embryo inoculation, hematocrit was lowered in both lines of chickens between 28 and 42 days of age. Immune response to sheep red blood cells and Brucella abortus was only slightly and inconsistently reduced by inoculation of RAV-1 by both routes in both lines of chickens. It was concluded that the major influences of ev gene expression was to reduce the incidence of NNS and the neutralizing antibody response after RAV-1 inoculation of day-old chicks. PMID- 2986086 TI - Effects of supplementary vitamin D3 on egg production of two strains of Japanese quail and growth of their progeny. AB - Two strains (University of Saskatchewan, white egg and wild type) of mature Japanese quail (Coturnix coturnix japonica), following a 4-week depletion period, were fed one of four diets varying in supplementary vitamin D3: a) no supplementary vitamin D3, b) National Research Council (NRC, 1977) recommended level of 1,200 IU vitamin D3/kg feed, c) 10 times, NRC levels, and d) 100 times NRC levels for 4 weeks. Egg production, egg weight, number of softshelled eggs, body weight, and feed consumption were not affected by vitamin D3 treatment. Hatchability of the white egg strain was not affected by vitamin D3 treatment. Hatchability in the wild type strain was reduced in those birds consuming diets unsupplemented with vitamin D3 although not significantly. Tibia ash was not affected by vitamin D3 treatment. Progeny from hens receiving no supplemental vitamin D3 were smaller throughout a 5-week growth period; however, feed consumption and percentage tibia ash content were similar to those of progeny from the other treatments. Progeny fed 480 IU vitamin D3/kg feed were larger, had higher tibia ash, and similar feed consumption compared with those fed a diet with no added vitamin D3. PMID- 2986087 TI - Serum lipoproteins in chickens after administration of adrenocorticotropin or exposure to high temperature. AB - The effects of intramuscular injections of adrenocorticotropin (ACTH) or short term exposures to high environmental temperature (44 to 46 C) were determined on the relative amounts of lipoproteins in the serum of 6 to 7-week-old White Rock chickens. In addition, the influence of the sex of the bird and fasting vs. ad libitum feeding on these lipoproteins was measured. The relative amount of high density lipoprotein (HDL) was lowered by ACTH, and heat exposure lowered both HDL and very low density lipoprotein (VLDL). Fasting completely eliminated VLDL and lowered low density lipoprotein (LDL) but had little effect on HDL. The temperature treatments significantly increased serum corticosteroid levels in both sexes in birds fed ad libitum but only in the males of fasted birds. Fasting for 15 hr did not significantly affect serum corticosteroid levels. PMID- 2986088 TI - Immunosuppressive effects of virulent strain of hemorrhagic enteritis virus in turkeys vaccinated against Newcastle disease. AB - One week after infection with a virulent strain of hemorrhagic enteritis virus (HEV), turkeys were vaccinated for Newcastle disease. The effect of a virulent strain of HEV on turkeys' immune response to Newcastle disease vaccine and the mitogenic response of their whole blood peripheral lymphocytes were examined. The results revealed a statistically significant difference (P less than .01) in the Newcastle disease hemagglutination inhibition (NDHI) antibody titers from turkeys infected with virulent HEV. The NDHI antibody titers were lower in turkeys exposed to virulent HEV before vaccination. There was an initial depression in phytohemagglutinin (PHA) response 1 week postinfection in turkeys infected with virulent HEV strain. PMID- 2986089 TI - Parabiosis between avian embryos resistant and susceptible to Rous sarcoma virus. AB - Parabiosis of chicken embryos was used to determine if resistance or susceptibility to Rous sarcoma virus (RSV) tumors could be transferred between RSV-resistant (R) and RSV-susceptible (S) embryos. Eggs from RSV-R and RSV-S lines were parabiosed at 12 days of incubation. One week after hatching the chicks were inoculated in the wing web with RSV. Birds were examined for tumor lesions through 9 weeks of age. Lesion scores of the nonparabiosed RSV-S line were significantly higher than nonparabiosed RSV-R lines. Lesion scores of RSV-R chicks that had been parabiosed to RSV-S chicks were higher than nonparabiosed RSV-R chicks. Lesion scores of RSV-S chicks parabiosed to RSV-R chicks were not different from nonparabiosed RSV-S chicks. These results suggest that the resistance to RSV was not transferred between parabionts but that there was significant transfer of susceptibility from the RSV-S embryos to the RSV-R embryos. PMID- 2986090 TI - Iatrogenic hypokalaemia. PMID- 2986092 TI - [Peripheral neuropathy during treatment with nadoxolol]. PMID- 2986091 TI - [Corticotropin releasing factor]. AB - The search for a neurohormone specifically controlling ACTH secretion resulted in the discovery of the corticotropin-releasing factor (CRF). This factor, located mainly in a paraventricular-infundibular hypothalamic tract, stimulates ACTH synthesis and secretion through a cAMP-dependent mechanism. The corticotropin releasing factor is the predominant component of a complex control system of adrenal cortex secretion, which also includes catecholamines and the antidiuretic hormone. Its specificity as stimulant of the corticotropic function makes it an extremely useful tool for physiological and physiopathological studies of the hypothalamus-pituitary-adrenal cortex axis regulation. PMID- 2986093 TI - [Development of cardiac adenylate cyclase hypersensitivity to isoproterenol in the chronic action of propranolol on rats]. AB - The chronic treatment of rats with the beta-adrenergic antagonist propranolol causes a double increase in the amount of beta-adrenergic receptors in the cardiac membranes. The purpose of the paper is to investigate the effect of propranolol on the activity and regulatory properties of rat heart adenylate cyclase. Propranolol injections to rats for 3 weeks (10-20 mg/l kg bw) did not influence the enzyme basal activity but caused a rise of a degree of myocardial adenylate cyclase activation by isoproterenol and glucagon. PMID- 2986094 TI - Cyclosporine-associated lymphoproliferation, despite controlled cyclosporine blood concentrations, in a renal allograft recipient. AB - In a patient receiving sulfinpyrazone (Anturan, Geigy) an unusually high dose of cyclosporine (Cys) was required to maintain serum values in the range of 50-200 ng/ml. After eight months of 1300-1500 mg/day, the patient complained of increasing malaise and symptoms of cyclosporine side-effects. This clinical state was accompanied by splenomegaly and two monoclonal peaks in the gamma region on serum electrophoresis. Concomitantly, rising cytomegalovirus IgM titres, following by rising IgG titres, indicated a primary cytomegalovirus infection. This ominous biclonal proliferation markedly diminished during the subsequent six months, during which time the cyclosporine dose was minimised. He returned to good health, splenomegaly and monoclonal gamma globulin virtually disappearing. He remains well at 16 months post-transplantation. PMID- 2986095 TI - Impaired regulation of beta-adrenoceptors in patients on maintenance haemodialysis. AB - In patients on maintenance haemodialysis the number of lymphocyte beta 2 adrenoceptors (determined by (+/-)-125 iodocyanopindolol binding) was not different from that in healthy controls; lymphocyte cyclic AMP responses to (-) isoprenaline (10(-8)-10(-4) M) or NaF (10 and 50 mM), however, were significantly reduced. Dynamic exercise on a bicycle (80% of maximum heart rate) for 15 minutes caused in 10 healthy volunteers a fourfold increase in plasma catecholamines; concomitantly lymphocyte beta 2-adrenoceptor number increased by about 55 per cent. In contrast, in patients on maintenance haemodialysis exercise induced only a twofold increase in plasma catecholamines and did not affect beta 2 adrenoceptor number. It is concluded that in chronic uraemia regulation and responsiveness of beta-adrenoceptors is impaired. PMID- 2986096 TI - Absence of a beneficial haemodynamic effect of bicarbonate versus acetate haemodialysis. AB - The present study compares data on blood pressure and clinical tolerance, obtained consecutively in the same patients during acetate and bicarbonate haemodialysis. Twenty-one patients were followed over an equal period of acetate and bicarbonate dialysis, averaging more than 30 months per patient. Absolute and relative blood pressure changes were noted. Contrary to what often has been claimed previously, it is concluded from the present long-term study that, bicarbonate haemodialysis has no specific beneficial effect on blood pressure in stabilised chronic patients. As far as vomiting and nausea are concerned, clinical tolerance is, however, significantly better than for acetate haemodialysis. PMID- 2986097 TI - Effect of dialysate buffer on potassium removal during haemodialysis. AB - There is a linear relationship between potassium removal during haemodialysis and plasma potassium (Kp). Kp falls rapidly during the first hour of dialysis but very little during the last two hours of a five hour dialysis. There is a fairly constant movement of potassium from the intracellular to extracellular space throughout dialysis. Total potassium removal is best predicted by pre-dialysis Kp, but change in Kp is related to the impact of dialysis on acid-base status. The choice of acetate or bicarbonate buffered dialysate does not effect potassium removal during dialysis. PMID- 2986098 TI - Favourable effects of bicarbonate dialysis on the body pool of phosphate. AB - An intravenous infusion of 3,430mg of PO4- has been given to 11 patients on acetate and to 11 patients on bicarbonate haemodialysis. The 'phosphate spaces' and dialytic removal were determined. The bicarbonate dialysis causes lower values of phosphate pool, total phosphate space, cellular space and phosphate cellular clearance. There is also a greater phosphate removal during bicarbonate dialysis. The better correction of metabolic acidosis and the absence of acetate metabolism are two factors which may be responsible for these phenomena. PMID- 2986099 TI - The cardiovascular and metabolic effects of mixtures of acetate and succinate: a potential improvement in dialysate solutions. AB - Various mixtures of acetate (AC) and succinate (SUCC) were studied for their metabolic and cardiovascular (CV) effects in 10 dogs. The CV effects seen with all mixtures were similar to those reported for SUCC alone with the only changes being increased cardiac output and decreased total peripheral resistance. The 50:50 per cent AC/SUCC offered the advantage of a rapid but less marked and more sustained HCO3 production. The pH changes in all mixtures followed HCO3 values. Since the addition of SUCC seems to reduce the untoward CV changes seen with AC alone, the ratio of AC/SUCC should be based on metabolic considerations. The present data suggest that a 50:50 per cent AC/SUCC mixture is optimal for metabolic and CV effects. PMID- 2986100 TI - Herpes virus infection prevalence in regular haemodialysis patients--a comparative evaluation of complement fixation, indirect immunofluorescence and Elisa tests. AB - The presence and titres of specific serum IgG and IgM antibodies to cytomegalovirus, herpes simplex virus and varicella-zoster virus were evaluated in 50 haemodialysis patients by complement fixation, immunofluorescence and Elisa tests. A second serum sample was tested in 24 patients after four weeks. Specific serum IgG antibodies to Epstein-Barr virus were also measured by immunofluorescence in 26 patients. By immunofluorescence and Elisa tests, the prevalence of cytomegalovirus, herpes simplex virus and Epstein-Barr virus infection is approximately 100 per cent, and varicella-zoster virus 60 per cent. High titres of IgG specific antibodies found by Elisa tests, detection of IgM antibodies in 16-18 per cent of patients and sero-conversion in 25 per cent of patients, suggests continuous antigenic stimulation. PMID- 2986101 TI - Computerised program to compare tolerance of dialysis patients to different dialysis schedules. AB - In the attempt to compare rapidly and easily the tolerance of dialysis patients to different dialysis schedules, we developed a computerised program for use with a personal computer. As a first application of this program we analysed the effects of long-term substitution of bicarbonate for acetate in reducing dialysis hypotension. PMID- 2986102 TI - Usefulness of 99mTc pyrophosphate bone scintigraphy in the survey of dialysis osteodystrophy. AB - Fogelman's score (FS) was used to determine the usefulness of 99mTc pyrophosphate (Tc-PP) bone scintigraphy in the evaluation of dialysis osteodystrophy. FS correlated well with bone 47Ca accretion rate. It remained stable after six months in patients treated with 1 alpha (OH)D3 and increased significantly in a randomised group of untreated patients. It decreased after two years of 1 alpha (OH)D3 therapy while serum calcium increased and iPTH and alkaline phosphatases decreased. Patients with low FS, treated by 1 alpha (OH)D3, rapidly developed hypercalcaemia. In cases of spontaneous hypercalcaemia, parathyroidectomy did not normalise serum calcium in patients with low FS despite a significant decrease in serum iPTH. Lower FS were associated with a higher increase in serum aluminium after desferrioxamine (DFO) administration and in two cases of proven aluminium osteomalacia, DFO therapy was followed by a dramatic increase in FS. PMID- 2986103 TI - Intracellular bicarbonate and pH of skeletal muscle in chronic renal failure. AB - In 11 controls and 10 patients suffering from untreated uraemic acidosis intracellular bicarbonate and skeletal muscle pH (needle biopsy) were determined. In all patients a significant intracellular acidosis, not related to any extracellular indices was found. It is concluded that the chronic proton load is able to effect intracellular buffer composition; moreover the action of other factors such as derangements of cell metabolism and nutritional imbalance could be operating. PMID- 2986104 TI - Pneumatosis intestinalis in patients after cadaveric kidney transplantation: possible relationship with an active cytomegalovirus infection. AB - Four patients are presented with pneumatosis intestinalis following kidney transplantation, all with severe cytomegalovirus (CMV) infection. Two patients had a primary infection and two patients had CMV reactivation. One patient died because of disseminated CMV infection. Two patients had concomitantly an active, non-obstructive duodenal ulcer. In a control population of 17 patients who suffered from a duodenal ulcer post-transplant without any evidence of CMV infection, we could not demonstrate pneumatosis intestinalis. We suggest a possible relationship between pneumatosis intestinalis and active CMV infection. The possible mechanisms responsible for this relationship are discussed. PMID- 2986105 TI - Two nonidentical forms of subunit V are functional in yeast cytochrome c oxidase. AB - In Saccharomyces cerevisiae, the inner mitochondrial membrane protein cytochrome c oxidase is composed of nine polypeptide subunits. Six of these subunits (IV, V, VI, VII, VIIa, VIII) are encoded by the nuclear genome, and the remaining three (I, II, III) are encoded by mitochondrial DNA. We report here the existence of two nonidentical subunit V polypeptides, which are encoded by separate genes within the yeast genome. One gene, COX5a, encodes the polypeptide Va, normally found in preparations of holocytochrome c oxidase. The other gene, COX5b, encodes the polypeptide Vb, which cross-reacts with anti-subunit Va antiserum and restores respiratory competency and cytochrome oxidase activity in transformants of cox5a structural gene mutants. This polypeptide also copurifies with the holoenzyme prepared from these transformants. We have found that COX5b is expressed in vegetatively growing yeast cells, and that the Vb polypeptide can be detected in mitochondria from strain JM28, a cox5a mutant. This mutant has 15% 20% residual cytochrome oxidase activity, and it respires at 10%-15% the wild type rate. By disrupting the COX5b gene in this strain, we show that this residual activity is directly attributable to the presence of a chromosomal copy of the COX5b gene. Taken together, these results suggest that Va or Vb can function as cytochrome oxidase subunits in yeast and that Vb may be used under some specific, as yet undefined, physiological conditions. PMID- 2986106 TI - The gene coding for carbamoyl-phosphate synthetase I was formed by fusion of an ancestral glutaminase gene and a synthetase gene. AB - A near full-length cDNA copy of rat carbamoyl-phosphate synthetase I (EC 6.3.4.16) mRNA has been cloned. The cDNA insert in the recombinant plasmid pHN234 is 5.3 kilobases long. Analysis of the sequence coding for carbamoyl-phosphate synthetase I indicates that the gene has arisen from a fusion of two ancestral genes: one homologous to Escherichia coli carA, coding for a glutaminase subunit, and the second homologous to the carB gene that codes for the synthetase subunit. A short amino acid sequence previously proposed to be part of the active site involved in glutamine amide nitrogen transfer in the E. coli and yeast carbamoyl phosphate synthetases (EC 6.3.5.5) is also present in the rat enzyme. In the mammalian enzyme, however, the glutaminase domain lacks a cysteine residue previously shown to interact with glutamine. The cysteine is replaced by a serine residue. This substitution could, in part, account for the inability of mammalian carbamoyl-phosphate synthetase I to catalyze the hydrolysis of glutamine to glutamic acid and ammonia. PMID- 2986107 TI - Adenovirus infection elevates levels of cellular topoisomerase I. AB - We have developed a specific, sensitive, and quantitative assay for topoisomerase I, which is based on the formation of a covalent enzyme-DNA intermediate. Our assay measures the quantitative transfer of 32P radioactivity from 32P-labeled DNA to topoisomerase I. Since 32P-labeled topoisomerase molecules are resolved by NaDodSO4/PAGE, HeLa topoisomerase I (100 kDa) and calf thymus topoisomerase I (82 kDa) can be quantitatively assayed in the same reaction mixture. The assay can detect at least 0.3 ng (3 fmol) of topoisomerase I. We have used our assay to measure the levels of topoisomerase I activity in crude extracts of nuclei prepared from uninfected, adenovirus-infected, and adenovirus-transformed human cells. The evidence suggests that an adenovirus early gene product, presumably a protein encoded in early region 1A (E1A), increases cellular topoisomerase I activity at least 10-fold. Immunoblotting analysis with antiserum against calf thymus topoisomerase I shows that the increase in activity is due to an increase in the amount of enzyme. PMID- 2986108 TI - The replication initiator protein of plasmid pSC101 is a transcriptional repressor of its own cistron. AB - The plasmid-encoded replication initiator protein of pSC101 specifically repressed initiation of transcription of its own cistron from its natural promoter. Addition of the purified initiator had little or no visible effect on transcription initiated from a heterologous promoter. DNase protection experiments revealed that the RNA polymerase recognition sequence was overlapped by the initiator protein recognition sequences, which are vicinal to the replication origin. Using the labeled promoter sequence, we have performed competitive DNase protection experiments in two ways: by adding RNA polymerase and initiator protein simultaneously or by sequentially adding first RNA polymerase and then initiator protein to the DNase reaction mixture. The RNA polymerase protection pattern was recessive to that of the initiator regardless of whether the two proteins were added simultaneously or sequentially. This observation suggests that the mechanism of autoregulation is due to competition of the two proteins for the sequences in and around the promoter region. Furthermore, the sequential addition experiments raise the possibility of displacement of RNA polymerase from the promoter by the initiator protein. PMID- 2986109 TI - Functional activation of the long terminal repeat of human T-cell leukemia virus type I by a trans-acting factor. AB - Promoter function for gene expression of the long terminal repeat (LTR) of human T-cell leukemia virus type I (HTLV-I) was studied by constructing plasmids containing the LTR sequence. The gene encoding chloramphenicol acetyltransferase (CATase) was linked to an HTLV-I LTR sequence (pLTR-CAT) by replacing the simian virus 40 promoter in plasmid pSV2-CAT with the LTR sequence. The transient CATase activities of cells transfected with the plasmids were compared. The results are summarized as follows: The HTLV LTR was active even in an epithelial cell line, with efficiency similar to that of the simian virus 40 promoter. pLTR-CAT expressed high CATase activity, 40-200 times that expressed by pSV2-CAT, in HTLV I-infected T-cell lines, such as the human cell lines MT-2 and HUT-102, or in HTLV-I-infected rat cell lines. This enhanced activity of the LTR seems to be associated with HTLV gene expression, since only low activity of pLTR-CAT was observed in the HTLV-infected cell line MT-1, in which only a small percent of cells express viral antigens. In HTLV-infected rat cell lines, the pX-encoded protein p40x was the only viral protein detected. Thus, we suggest that p40x is the factor associated directly or indirectly with the enhanced activity of the LTR. PMID- 2986110 TI - c-erbB activation in avian leukosis virus-induced erythroblastosis: clustered integration sites and the arrangement of provirus in the c-erbB alleles. AB - There is considerable evidence that links the activation of cellular genes to oncogenesis. We previously reported that structural rearrangements in the cellular oncogene c-erbB correlate with the development of erythroblastosis induced by avian leukosis virus (ALV). c-erbB recently has been shown to be related to the gene encoding epidermal growth factor receptor. We now have characterized the detailed mechanisms of c-erbB activation by ALV proviruses. We report here that the ALV proviral integration sites are clustered 5' to the region where homology to v-erbB starts, suggesting that interruption in this region of c-erbB is important for its activation. The proviruses are oriented in the same transcriptional direction as c-erbB and usually are full-size. The latter finding is in contrast to the frequent deletions observed within the c-myc linked proviruses in B-cell lymphomas. We have also identified a second c-erbB allele, which differs from the previously known allele primarily by a deletion in an intron region. This allele is also oncogenic upon mutation by an ALV provirus. PMID- 2986111 TI - Identification and functional characterization of leukotriene B4 20-hydroxylase of human polymorphonuclear leukocytes. AB - A single reaction product was formed during the incubation of 1.5 microM (5S,12R) dihydroxy-6,14-cis-8,10-trans-[3H]icosatetraenoic acid (leukotriene B4, LTB4) for 30 min at 37 degrees C in 10 mM potassium phosphate buffer (pH 7.5) with 100 microM NADPH and the 150,000 X g supernatant of sonicated human polymorphonuclear leukocytes (PMN). The reaction product exhibited the same mobility on reversed phase HPLC (RP-HPLC) and TLC as standard 20-hydroxy-LTB4 (20-OH-LTB4). When the omega-oxidation product of [3H]LTB4 was eluted from a Sep-Pak, resolved by RP HPLC, and analyzed by GC/MS, its structure was determined to be solely 20-OH LTB4. The Km of the 20-hydroxylase for [3H]LTB4 at its optimal pH of 7.5 was 0.22 +/- 0.08 microM (mean +/- SD, n = 4) and the Vmax was 48 +/- 11 pmol/min X mg of protein (mean +/- SD, n = 4). When the concentration of [3H]LTB4 was fixed at 1.5 microM, the Km for NADPH was 1.01 +/- 0.59 microM (mean +/- SD, n = 3). The location in the 150,000 X g supernatant of the LTB4 20-hydroxylase distinguishes it from the cytochrome P-450 system of liver, lung, and kidney microsomes and from the NADPH oxidase-cytochrome b-245 system of the human PMN. The LTB4 20 hydroxylase is either a unique cytochrome P-450 or other monooxygenase. PMID- 2986112 TI - Identification of cDNA clones encoding a precursor of rat liver cathepsin B. AB - Recent studies have suggested that many lysosomal enzymes, including cathepsin B (EC 3.4.22.1), may be synthesized as larger precursors and proteolytically processed to their mature forms. To determine the structure of the primary translation product of cathepsin B, we have screened a phage cDNA library for clones encoding rat liver cathepsin B. We synthesized two extended DNA oligonucleotides to use as hybridization probes: a 50-mer corresponding to the coding segment for residues 215-231 of mature cathepsin B and a 54-mer corresponding to residues 117-134. After screening 600,000 plaques, five clones were obtained that hybridized to the 32P-labeled 50-mer; of these, two (lambda rCB3 and lambda rCB5) also reacted with the 54-mer. DNA sequence analysis confirmed that lambda rCB3 and lambda rCB5 both encoded rat liver cathepsin B, and the translated sequence is in agreement with the sequence determined [Takio, K., Towatari, T., Katunuma, N., Teller, D. C. & Titani, K. (1983) Proc. Natl. Acad. Sci. USA 80, 3666-3670], except for a tryptophan for glycine substitution at residue 78 and the presence of two amino acids at the junction site of the light and heavy chains. Moreover, the DNA sequence reveals an open reading frame extending beyond the 5' (NH2 terminus), and the predicted COOH terminus of the coding sequence for the mature protein is extended by six amino acids. These results confirm that the biosynthesis of cathepsin B involves a larger precursor form and demonstrate the effectiveness of long oligonucleotide probes for screening to detect rare cloned mRNAs. PMID- 2986113 TI - Evolution and organization of the fibrinogen locus on chromosome 4: gene duplication accompanied by transposition and inversion. AB - Human fibrinogen cDNA probes for the alpha-, beta-, and gamma-polypeptide chains have been used to isolate the corresponding genes from human genomic libraries. There is a single copy of each gene. Restriction endonuclease analysis of isolated genomic clones and human genomic DNA indicates that the human alpha-, beta-, and gamma-fibrinogen genes are closely linked in a 50-kilobase region of a single human chromosome: the alpha-gene in the middle flanked by the beta-gene on one side and the gamma-gene on the other. The alpha- and gamma-chain genes are oriented in tandem and transcribed toward the beta-chain gene. The beta-chain gene is transcribed from the opposite DNA strand toward the gamma- and alpha chain genes. The three genes have been localized to the distal third of the long arm of chromosome 4, bands q23-q32, by in situ hybridization with fibrinogen cDNAs and by examination of DNA from multiple rodent-human somatic cell hybrids. Alternative explanations for the present arrangement of the three fibrinogen genes involve either a three-step mechanism with inversion of the alpha/gamma region or a two-step mechanism involving remote transposition and inversion. The second more simple mechanism has a precedent in the origin of repeated regions of the fibrinogen and immunoglobulin genes. PMID- 2986114 TI - Ultraviolet resonance Raman excitation profiles of nucleic acid bases with excitation from 200 to 300 nanometers. AB - Raman spectra are presented for dilute aqueous solutions of the four ribonucleotides AMP, GMP, UMP, and CMP obtained with laser excitation at 299, 266, 253, 240, 229, 218, 209, and 200 nm. Distinct evidence of strong, selective resonance enhancement is obtained. Low-resolution excitation profiles have been constructed for the strongest bands by using the phosphate band at 994 cm-1 as an internal reference. The excitation spectra for many of the vibrational bands are dominated by a peak corresponding to the lowest-energy electronic transition near 260 nm. Smaller peaks are seen for higher-energy electronic transitions. For some modes, the resonance enhancement is dominated by the higher-energy transitions. It is clear from these new data that a full description of the resonance Raman profiles of the nucleic acids will have to include several excited electronic states. Two examples are given of cases where ionic species can be distinguished easily by using far-UV excitation, but these species are indistinguishable with 266-nm excitation. This demonstrates the utility of far-UV resonance Raman spectroscopy for obtaining structural information. PMID- 2986115 TI - Specific expression of the human cellular fps/fes-encoded protein NCP92 in normal and leukemic myeloid cells. AB - We have found that both an antibody directed against a synthetic peptide representing an amino acid sequence of the conserved kinase domain of transforming protein P140 of Fujinami sarcoma virus and a regressing tumor antiserum recognized the products of the c-fps/fes genes of both avian and mammalian cells. The anti-peptide antibody also recognized a 94-kilodalton protein that was related to but distinct from the c-fps/fes product in structure and in tissue distribution. A 92-kilodalton protein, NCP92, was found to be the mammalian counterpart of the previously identified avian c-fps/fes protein NCP98 by its structural similarity to NCP98, its associated tyrosine kinase activity, and its similar tissue distribution. The highest levels of NCP92 were found in tissue macrophages and in bone marrow. In bone marrow NCP92 expression was restricted to cells of the monocyte/macrophage and granulocyte lineages. That the expression of NCP92 is limited to these cell types was confirmed by the analysis of murine and human hematopoietic tumors representing different cell lineages: NCP92 was positive in leukemic cells of granulocytic and monocytic origin but not in B-lymphocytic, T-lymphocytic, or erythroid tumor cells. The expression of NCP92 seems to be related to the capacity of myeloid cells to differentiate and to respond to certain colony-stimulating factors. PMID- 2986116 TI - Reduction of gap junctional conductance by microinjection of antibodies against the 27-kDa liver gap junction polypeptide. AB - Antibody raised against isolated rat liver gap junctions was microinjected into coupled cells in culture to assess its influence on gap junctional conductance. A rapid inhibition of fluorescent dye transfer and electrical coupling was produced in pairs of freshly dissociated adult rat hepatocytes and myocardial cells as well as in pairs of superior cervical ganglion neurons from neonatal rats cultured under conditions in which electrotonic synapses form. The antibodies have been shown by indirect immunofluorescence to bind to punctate regions of the plasma membrane in liver. By immunoreplica analysis of rat liver homogenates, plasma membranes, and isolated gap junctions resolved on NaDodSO4/polyacrylamide gels, binding was shown to be specific for the 27-kDa major polypeptide of gap junctions. This and similar antibodies should provide a tool for further investigation of the role of cell-cell communication mediated by gap junctions and indicate that immunologically similar polypeptides comprise gap junctions in adult mammalian cells derived from all three germ layers. PMID- 2986117 TI - Natural killer cell recognition of target cells expressing different antigens of vesicular stomatitis virus. AB - Natural killer (NK) cells have the capability of lysing virus-infected, transformed, and embryonal cells, yet the nature of the target structure(s) recognized remains unclear. The availability of well-characterized temperature sensitive (ts) mutants of vesicular stomatitis virus, defective in expression of individual viral-encoded polypeptides at the nonpermissive temperature (39 degrees C), offered an approach to elucidating NK-cell recognition of virus infected cells. Target cells were infected with ts mutants in three functions: the viral surface glycoprotein (G protein; ts 045); the matrix (M) protein (ts G31, ts G33), and the polymerase (ts G11). Cells infected with wild-type virus and all ts mutants at the permissive temperature (31 degrees C) were killed by murine spleen cells. Similar to results on cytotoxic T lymphocytes, target cells infected by ts 045 defective in expression of G protein at 39 degrees C were not killed by NK cells. Unexpectedly, cells infected at 39 degrees C with the M protein mutants also were not killed, although G protein was expressed at the cell surface. Target binding studies indicated that conjugates were not formed by cells infected with the ts mutants at the nonpermissive temperature. That expression of G protein was not sufficient for NK cell-mediated cytotoxicity was established in experiments in which a plasmid (pSVGL) containing the gene for vesicular stomatitis virus G protein was transfected into COS cells. Although G antigen was expressed on the plasma membrane, the cells were not lysed. These results suggest either that recognition of virus-infected cells depends on an appropriate conformation imparted to the viral G protein by association with the M protein or that NK cells can recognize alterations in the structure of the cell membrane induced by insertion of viral M and G molecules. PMID- 2986118 TI - Metabolic activation of the nucleoside analog 9-[( 2-hydroxy-1 (hydroxymethyl)ethoxy]methyl)guanine in human diploid fibroblasts infected with human cytomegalovirus. AB - 9-[( 2-Hydroxy-1-(hydroxymethyl)ethoxy]-methyl)guanine (BW B759U) is a more potent inhibitor of human cytomegalovirus (HCMV) in vitro than is the related nucleoside analog acyclovir (ACV). BW B759U was selectively activated to the 5' triphosphate (BW B759U-triphosphate) in cells infected with HCMV to levels at least 10-fold higher than those measured for ACV-triphosphate and up to as much as 100-fold higher than the levels found in uninfected cells. BW B759U triphosphate accumulated in HCMV-infected cells with time; the rate of this increase was dependent upon the drug dose and virus multiplicity of infection. Enzyme activities that catalyzed the phosphorylation of thymidine and 2' deoxycytidine increased 3- to 7-fold in extracts of cells early after HCMV infection but thereafter declined. No concomitant increase in the rate of BW B759U phosphorylation was detected under these assay conditions. Maximal rate of accumulation of both BW B759U-triphosphate and ACV-triphosphate after a short exposure to drug occurred in the late phase of the infective cycle, as the titer of extracellular virus reached a peak in untreated cultures, but after the decline of stimulated host deoxypyrimidine kinase activities. Once formed, the BW B759U-triphosphate pool decreased very slowly and thus it persisted for several days in both HCMV-infected and uninfected cells. PMID- 2986119 TI - Plasma and cellular retinoid-binding proteins and transthyretin (prealbumin) are all localized in the islets of Langerhans in the rat. AB - The immunohistochemical localization of plasma retinol-binding protein (RBP), cellular retinol-binding protein (CRBP), and transthyretin (TTR) was studied in rat pancreas. The studies employed antibodies purified by immunosorbent affinity chromatography, permitting the specific staining and localization of each antigen by the unlabeled peroxidase-antiperoxidase method. Specific immunostaining for each of these three proteins was found localized to the islets of Langerhans. Both RBP and CRBP were localized in cells that were peripherally distributed within the islets, with an anatomic distribution that resembled that of the glucagon-containing A cells. Immunoreactive TTR was localized in cells that were more centrally distributed in the islets, with an anatomic distribution that resembled that of the insulin-containing B cells. These findings were confirmed by radioimmunoassay of a homogenate of isolated rat islets. By using sensitive and specific radioimmunoassays for each antigen, unusually high levels of CRBP, RBP, TTR, and cellular retinoic acid-binding protein (CRABP) were found in rat islets. The physiological significance of the localization of RBP, CRBP, CRABP, and TTR in the islets is not known. The findings suggest that retinoids and their binding proteins may play important metabolic roles within islet cells, and hence that they may be involved in some way in the biological, endocrine, function of the islets. PMID- 2986120 TI - Characterization of [3H][2-D-penicillamine, 5-D-penicillamine]-enkephalin binding to delta opiate receptors in the rat brain and neuroblastoma--glioma hybrid cell line (NG 108-15). AB - Specific binding properties of the tritium-labeled delta opiate receptor agonist [3H][2-D-penicillamine, 5-D-penicillamine]enkephalin [( 3H][D-Pen2, D Pen5]enkephalin) were characterized in the rat brain and in a mouse neuroblastoma rat glioma hybrid cell line (NG 108-15). Saturation isotherms of [3H][D-Pen2, D Pen5]enkephalin binding to rat brain and NG 108-15 membranes gave apparent Kd values of 1-6 nM. These values are in good agreement with the Kd value obtained from the kinetic studies. The Bmax value in NG 108-15 membranes was 235.3 fmol/mg of protein. An apparent regional distribution of [3H][D-Pen2, D-Pen5]enkephalin binding was observed in the rat brain. A number of enkephalin analogues inhibited [3H][D-Pen2, D-Pen5]enkephalin binding with high affinity (IC50 values of 0.5-5.0 nM) in both NG 108-15 and rat brain membranes. However, putative mu receptor selective ligands such as morphine, [D-Ala2, MePhe4, Gly5-ol]enkephalin, [MePhe3, D-Pro4]morphiceptin, and naloxone were less effective inhibitors of [3H][D-Pen2, D-Pen5]enkephalin binding in both systems tested. These data suggest that [3H][D Pen2, D-Pen5]enkephalin is a potent and selective ligand for the delta opioid receptor. PMID- 2986121 TI - Associative conditioning analog selectively increases cAMP levels of tail sensory neurons in Aplysia. AB - Bilateral clusters of sensory neurons in the pleural ganglia of Aplysia contain cells involved in a defensive tail withdrawal reflex. These cells exhibit heterosynaptic facilitation in response to noxious skin stimulation that can be mimicked by the application of serotonin. Recently it has been shown that this facilitation can be selectively amplified by the application of a classical conditioning procedure to individual sensory neurons. We now report that an analog of this classical conditioning paradigm produces a selective amplification of the cAMP content of isolated sensory neuron clusters. The enhancement is achieved within a single trial and appears to be localized to the sensory neurons. These results indicate that a pairing-specific enhancement of cAMP levels may be a biochemical mechanism for associative neuronal modifications and perhaps learning. PMID- 2986122 TI - Sequence-specific recognition of B-DNA by oligo(N-methylpyrrolecarboxamide)s. AB - Four homologous oligopeptide-EDTA molecules, tri-, tetra, penta-, and hexa(N methylpyrrolecarboxamide)-EDTA, in the presence of Fe(II), O2, and dithiothreitol, cleave 32P-end-labeled restriction fragments from plasmid pBR322 DNA at common locations rich in A X T base pairs that differ in the size of the binding site. From analysis of the cleavage patterns visualized by high resolution denaturing gel electrophoresis, the oligopeptides with three, four, five, and six N-methylpyrrolecarboxamide units, containing four, five, six, and seven amide NHs, bind sites of A X T-rich DNA consisting of five, six, seven, and eight contiguous base pairs, respectively. The general rule of n amides affording binding site sizes of n + 1 base pairs is consistent with the oligopeptides binding in the minor groove of right-handed DNA, with the amide NH groups forming bridges between the adjacent N-3 and O-2 atoms of adenine or thymine on opposite strands of the DNA helix. PMID- 2986123 TI - Direct measurement of the electrogenic activity of o-type cytochrome oxidase from Escherichia coli reconstituted into planar lipid bilayers. AB - Turnover of o-type cytochrome oxidase purified from Escherichia coli and reconstituted into proteoliposomes leads to the generation of a transmembrane electrical potential (interior negative) by means of vectorial electron flow. In the experiments reported here, purified oxidase is reconstituted in planar lipid bilayers formed at the tip of patch pipets, and open-circuit membrane potentials generated by electron transfer are measured directly. Potentials of up to 4 mV (substrate side positive) are generated in the presence of reduced phenazine methosulfate or ubiquinol-1, and with both substrates, electrogenic activity is inhibited by cyanide. Furthermore, the membrane potential generated during oxidase turnover is inhibited progressively with applied voltages (substrate side positive), decreasing almost to zero at an applied voltage of 150 mV. PMID- 2986124 TI - Differential regulation of bovine brain calmodulin-dependent cyclic nucleotide phosphodiesterase isoenzymes by cyclic AMP-dependent protein kinase and calmodulin-dependent phosphatase. AB - Purified bovine brain calmodulin-dependent cyclic nucleotide phosphodiesterase (3',5'-cyclic-nucleotide 5'-nucleotidohydrolase, EC 3.1.4.17) contains isozymes that are composed of two distinct subunits with molecular masses of 60,000 and 63,000 daltons. Analysis by NaDodSO4 gel electrophoresis and autoradiography of a phosphodiesterase sample phosphorylated in the presence of [32P]ATP and bovine heart cAMP-dependent protein kinase catalytic subunit revealed that only the 60 kDa subunit was phosphorylated. By using an isozyme preparation greatly enriched with the 60-kDa subunit, the following observations regarding the subunit phosphorylation were made. First, the phosphorylation resulted in the maximal incorporation of about 2 mol of phosphate per mol of subunit. Second, complete inhibition of 60-kDa subunit phosphorylation was approached at a saturating concentration of Ca2+ when a molar ratio of calmodulin to phosphodiesterase of 2:1 was used. No inhibition was observed in the presence of either Ca2+ or calmodulin alone. Third, the phosphorylation was accompanied by a decrease in the enzyme affinity for calmodulin; calmodulin concentrations required for 50% activation of nonphosphorylated and maximally phosphorylated phosphodiesterase isozyme samples were 0.51 and 9.3 nM, respectively. Fourth, the phosphodiesterase isozyme could be dephosphorylated by the calmodulin-dependent phosphatase (calcineurin) in the presence of Ni2+ or Mn2+, the dephosphorylation being associated with an increase in the enzyme affinity for calmodulin. Fifth, peak II rabbit liver phosphoprotein phosphatase catalytic unit did not catalyze the dephosphorylation of the phosphodiesterase isozyme. PMID- 2986125 TI - Sequence variation in the gene for the immunogenic capsid protein VP1 of foot-and mouth disease virus type A. AB - The nucleotide sequences have been determined and compared from cloned cDNA genes coding for the foot-and-mouth disease virus (FMDV) immunogenic capsid protein, VP1, from eight different A subtypes: A5 Westerwald/58, A12 119ab (large plaque variant), A22 550 USSR/65, A24 Cruzeiro Brazil/55, A27 Cundinamarca Colombia/76, A32 Venezuela/70, A Venceslau Brazil/76, and A Argentina/79. We have also found sequence variations among different cDNA clones of the A5 and A24 subtypes. There are regions of nucleotide sequence within the VP1 gene that vary considerably among the subtypes as well as other regions that remain relatively constant. One highly variable region (codons 130-171) encodes amino acids previously identified as being exposed on the virus surface and constituting an important immunogenic site of the virus. There potentially exist secondary structures within the viral RNA sequences that code for this immunogenic site that could decrease the fidelity of replication at this sequence. The rapid generation of FMDV variants encouraged by such structures in the RNA could work together with various selective pressures to explain the observed accumulation of immunologically distinct viruses of the FMDV A type. PMID- 2986126 TI - 1,2-Diacylglycerol and phorbol ester inhibit agonist-induced formation of inositol phosphates in human platelets: possible implications for negative feedback regulation of inositol phospholipid hydrolysis. AB - The present study has demonstrated that pretreatment of human platelets with either phorbol ester or 1,2-diacylglycerol inhibits agonist-induced formation of inositol phosphates; this inhibition can be correlated with a decrease in the release of ATP and 5-hydroxytryptamine by thrombin. The mechanism of this action is not known, but a role for protein kinase C is suggested, as both phorbol ester and 1,2-diacylglycerol have in common the ability to activate this enzyme. These results have important implications as a possible negative feedback control over agonist-induced hydrolysis of inositol phospholipids. PMID- 2986127 TI - Primary structure and gene organization of human hepatitis A virus. AB - The RNA genome of human hepatitis A virus (HAV) was molecularly cloned. Recombinant DNA clones representing the entire HAV RNA were used to determine the primary structure of the viral genome. The length of the viral genome is 7478 nucleotides. An open reading frame starting at nucleotide 734 and terminating at nucleotide 7415 encodes a polyprotein of Mr 251,940. Comparison of the HAV nucleotide sequence with that of other picornaviruses has failed to reveal detectable areas of homology. However, a computer analysis of the putative amino acid sequence of HAV and poliovirus demonstrated the existence of short areas of homology in virion protein 3 (VP3) and throughout the carboxyl-terminal portion of the polyproteins. In addition, extensive protein structural homologies with poliovirus were detected. PMID- 2986128 TI - Identification of pTiC58 plasmid-encoded proteins for virulence in Agrobacterium tumefaciens. AB - Analyses were made of the host-dependent-variation (hdv) locus of the virulence (vir) region of the pTiC58 plasmid of Agrobacterium tumefaciens. The hdv locus is comprised of at least four genes that encode polypeptides of 13, 15, 29, and 28 kDa. Insertion of transposon Tn5 in the first gene abolishes the expression of all four genes in vitro and in vivo. Nucleotide sequence analysis of the hdv locus revealed four open reading frames tandemly arranged with spacer sequences having no promoter-like sequences and lacking the ability to bind A. tumefaciens RNA polymerase. These studies suggest that the hdv locus is comprised of at least four genes arranged in an operon in the vir region. The protein products of these genes are likely to function in some aspect of the host-range determination of A. tumefaciens. PMID- 2986129 TI - Cloning of Physarum actin sequences in an exonuclease-deficient bacterial host. AB - A genomic library of Physarum was constructed in the replacement vector EMBL3. Efficient propagation of the recombinant phages occurred only on the recBC-sbcB- host Escherichia coli CES200, which is deficient in the exonucleases I and V. Thirteen different recombinants with actin-related sequences were detected and 10 were purified from 90,000 plaques (the equivalent of 6 Physarum genomes) on strain CES200. Comparison of the plating efficiencies of the library and the actin-related isolates suggests that palindromic DNA sequences are responsible for the instability of Physarum DNA in E. coli. In one of these isolates, lambda PpA10, and in a 2.81-kilobase subclone of that isolate in plasmid pBR322, a deletion of 360 base pairs was detected that led to stable propagation of the recombinant DNA molecules in Rec+ E. coli. Electron microscopic analysis of the 2.81-kilobase fragment, after denaturation and self-hybridization, revealed secondary structures consistent with "foldback" structures. Restriction and DNA blot analysis of lambda PpA10 suggest that the unstable DNA segment is in close proximity to, if not part of, the previously defined actin-gene locus ardA. PMID- 2986130 TI - Molecular cloning of a full-length cDNA for human alcohol dehydrogenase. AB - We have cloned a full-length cDNA coding for human alcohol dehydrogenase (ADH; alcohol:NAD+ oxidoreductase, EC 1.1.1.1) from a human liver cDNA library constructed in phage lambda gt11. The library was screened by using a rabbit antibody against human ADH as a first probe, by the modified method of Young and Davis [Young, R. A. & Davis, R. W. (1983) Proc. Natl. Acad. Sci. USA 80, 1194 1198]. Mixed 14-mer synthetic oligonucleotides encoding Asp-Asp-His-Val-Val and Gln-Cys-Gly-Lys-Cys were used as a second probe. These amino acid sequences are considered to be common in all three subunits (alpha, beta, and gamma) controlled by the ADH1, ADH2, and ADH3 loci. Ten lambda gt11 recombinants of 35 positive plaques obtained by antibody screening contained inserted cDNAs of 1.5-2.4 kilobase pairs and were found to exhibit positive signals by hybridization with synthetic probes. One of them, with an inserted cDNA of 1631 base pairs, contained a sequence that encodes 374 amino acid residues of the human beta 1 subunit, a chain initiation codon, a chain termination codon, and additional 3' and 5' untranslated regions. A complete amino acid sequence of the human beta 1 subunit was deduced from the cDNA. PMID- 2986131 TI - Nucleotide sequence of two overlapping myc-related genes in avian carcinoma virus OK10 and their relation to the myc genes of other viruses and the cell. AB - Avian carcinoma virus OK10 has the genetic structure gag-delta pol-myc-delta env. It shares the transformation-specific myc sequence with three other avian carcinoma viruses (MC29, MH2, CMII) and also with a normal chicken gene proto-myc and the gag, pol, and env elements with non-transforming retroviruses. Unlike the other myc-containing viruses, which synthesize singular myc proteins, OK10 synthesizes two different myc-related proteins of 200 and 57 kDa. Here we have sequenced the myc region of an infectious OK10 provirus to investigate how OK10 synthesizes two different proteins from the same myc domain and to identify characteristic differences between the normal proto-myc gene and the myc-related viral transforming genes. It was found that the 1.6-kilobase myc domain of OK10 is colinear and coterminal with the myc domains of MC29, MH2, and the terminal two exons of proto-myc. It is preceded by the same splice acceptor as the myc sequence of MH2 and as the second proto-myc exon. From this and the known structure of retroviruses, it follows that the OK10 gene encoding the 57-kDa protein is discontinuous with a small 5' exon that includes six gag codons and a large 3' myc exon (delta gag-myc). This gene and the delta gag-myc gene of MH2 are isogenic. The proto-myc-derived intron preceding the myc domain of OK10 is in the same reading frame as the adjacent delta pol and myc domains and, hence, is part of the gag-delta pol-myc gene encoding the 200-kDa protein. Sequence comparisons with proto-myc and MC29 and MH2 indicate that there are no characteristic mutations that set apart the viral myc domains from proto-myc. It is concluded that transforming function of viral myc-related genes correlates with the lack of a viral equivalent of the first proto-myc exon(s) and conjugation of the viral myc domains with large or small retroviral genetic elements rather than with specific point mutations. Because OK10 and MH2 each contain two genes with potential transforming function (namely, delta gag-myc and gag-delta pol-myc or delta gag-mht, respectively), it remains to be determined whether the delta gag-myc genes have transforming function on their own or need helper genes. The possible helper requirement cannot be very specific because the two potential helper genes are very different. PMID- 2986133 TI - Modification of pK values caused by change in H-bond geometry. AB - The competition between various groups for a proton is studied by ab initio molecular orbital methods. It is found that reorientations of the two groups involved in a H-bond can reverse the equilibrium position of the proton shared between them. Specifically, the carbonyl and hydroxyl groups were modeled by H2CO and HOH. In the H-bond between these two groups, association of the proton with the carbonyl (H2COH...OH2)+ is favored over the hydroxyl (H2CO...HOH2)+ when the latter group is situated along a lone pair of the carbonyl oxygen. However, displacement of the water to the C = O axis between the two carbonyl lone pairs reverses the situation and (H2CO...HOH2)+ is more stable. A similar reversal of stability is observed in the H-bond involving a Schiff base (modeled by CH2NH) and amine (NH3). In one arrangement where the lone pairs of the two groups point toward one another, the proton prefers the Schiff base to the amine--i.e., (H2CHNH...NH3)+ is more stable than (H2CHN...HNH3)+. On the other hand, rotation of the lone pair of the amine away from the Schiff base nitrogen results in proton transfer across to the amine. These shifts in stability correspond to reversal of relative pK of the groups involved. A fundamental principle emerging from the calculations is that ion-dipole electrostatic interactions favor transfer of a proton to the group that is positioned as closely as possible to the negative end of the dipole moment vector of the other. The ideas developed here suggest a number of means by which conformational changes may be utilized to shift protons from residue to residue within a protein molecule such as an enzyme or bacteriorhodopsin. PMID- 2986132 TI - Poliovirus protease does not mediate cleavage of the 220,000-Da component of the cap binding protein complex. AB - Poliovirus infection of HeLa cells results in a rapid shutoff of host protein synthesis but does not inhibit the translation of poliovirus mRNA. It has been suggested that this virus-induced translational control is mediated by the inactivation of a cap binding protein (CBP) complex, and it has been shown that the 220,000-Da component(s) (p220) of the CBP complex is cleaved in infected HeLa cells to form antigenically related peptides of 100,000-130,000 Da. To determine whether the known viral protease (peptide 3C) was the mediator of the cleavage of p220, we used immunoblot techniques to analyze partially purified infected HeLa cell extracts for cleavage activity. We report here that p220 cleavage activity does not copurify with viral peptide 3C or with any precursors containing 3C sequences. We also show that cleavage of p220 can be demonstrated in vitro in HeLa cell extracts under conditions where the functional activity of the poliovirus protease is inhibited by specific antibody. PMID- 2986134 TI - Generation of leukotrienes by human monocytes upon stimulation of their beta glucan receptor during phagocytosis. AB - Human monocytes possess a receptor for ingestion of particulate activators of the human alternative complement pathway that functions in the absence of plasma proteins and is distinct from the receptors for Fc-IgG and the major cleavage fragment of the third component of complement (C3b). Incubation of monolayers of monocytes with 1.1 X 10(6) to 2.2 X 10(7) glucan particles per ml initiated a phagocytic response comparable to that obtained with zymosan particles, of which beta-glucan is a constituent along with mannan. Maximal quantities of 4.93 +/- 3.43 ng of leukotriene B4 (LTB4) and 0.43 +/- 0.23 ng of leukotriene C4 (LTC4) (mean +/- SD, n = 3) were released by 10(6) monocytes stimulated with 1.1 X 10(7) glucan particles per ml. Preincubation of monocytes with 50 micrograms of soluble beta-glucan per ml reduced subsequent monocyte ingestion of 5 X 10(6) zymosan particles per ml and 2.2 X 10(6) glucan particles per ml by 52% and 55%, respectively, and diminished release of LTB4 by monocytes stimulated with 2 X 10(8) zymosan particles per ml and 8.6 X 10(6) glucan particles per ml by 73% and 61%, respectively. Preincubation with 1 mg of soluble mannan per ml had little effect on monocyte phagocytosis or LTB4 generation in response to either zymosan or glucan particles, and neither soluble beta-glucan nor mannan stimulated generation of LTB4 or LTC4. The effect of pretreatment of monocytes with soluble beta-glucan was time dependent, with the maximal effect being evident within 20 min of pretreatment, and was specific for zymosan or glucan particles in that the LTB4 and LTC4 release induced by 2.5 microM calcium ionophore A23187 was unaffected. That both phagocytosis and leukotriene generation are inhibited by soluble beta-glucan but not by mannan at a rate compatible with the phagocytic process of monocyte monolayers indicates ligand specificity for a beta-glucan receptor. As the beta-glucan receptor recognizes particulate activators of the alternative complement pathway, the nonimmune response to a single stimulus induces complement activation, phagocytosis, and leukotriene generation. PMID- 2986136 TI - Purification of hemopoietin 1: a multilineage hemopoietic growth factor. AB - Hemopoietin 1 (H-1) and the mononuclear phagocyte specific growth factor CSF-1 act synergistically on developmentally early bone marrow cells to generate primitive CSF-1 receptor-bearing cells. The H-1 activity of the serum-free medium conditioned by the human urinary bladder carcinoma cell line 5637 was shown to result from the sum of the activities of two charged species (pI approximately equal to 4.8, approximately equal to 85%; pI approximately equal to 5.3, approximately equal to 15%) of similar size. No qualitative difference in the biological activity of these two species was detected. A four-step procedure, involving batch DEAE-cellulose chromatography, chromatofocusing, gel filtration, and hydrophobic chromatography has been developed for the major (pI approximately equal to 4.8) species. H-1 was purified approximately 65,000-fold and recovered as 32% of the total activity of the starting material. The lowest concentration yielding maximal biological activity was approximately equal to 0.25 ng/ml. The 125I-labeled purified H-1, in either native or reduced form, behaved as a homogeneous single band that coelectrophoresed with the biological activity of purified H-1 on sodium dodecyl sulfate gel electrophoresis (NaDodSO4/PAGE). The molecular mass of the purified reduced H-1, determined by NaDodSO4/PAGE was approximately equal to 17 kDa. Recent studies indicate that the purified H-1 is a multilineage hemopoietic growth factor. PMID- 2986135 TI - Avian myelocytomatosis virus immortalizes differentiated quail chondrocytes. AB - Quail embryo chondrocytes in culture display two morphological phenotypes: polygonal epithelial-like and floating cells. Both cell populations synthesize cartilage extracellular matrix proteins (type II collagen and specific proteoglycans), whereas type X collagen, which appears to be a marker of later stages of chondrocyte differentiation, is expressed only by the epithelial-like cells. Avian myelocytomatosis virus strain MC29 does not induce morphological transformation in quail embryo chondrocytes but stimulates these cells to proliferate with a progressively reduced doubling time. MC29-infected chondrocytes can be established in culture as a continuous cell line, whereas control (uninfected) cultures only survive a few months. Rapidly dividing MC29 infected chondrocytes still express type II collagen and cartilage proteoglycans but do not synthesize type X collagen. PMID- 2986137 TI - Platelet-derived collagenase inhibitor: characterization and subcellular localization. AB - Purified human platelets were found to contain a collagenase inhibitor that is immunologically, functionally, and chromatographically identical to that produced by human skin fibroblasts. None of the other formed elements of the blood (erythrocytes, granulocytes, mononuclear cells) possessed detectable quantities of this protein. Virtually all the collagenase inhibitor contained within platelets was released following platelet activation with thrombin. Similarly, platelet activation accompanying blood clotting also resulted in the release of this protein, the ratio of plasma to serum inhibitor levels being approximately equal to 0.5. When platelets were subjected to subcellular fractionation, essentially all of the platelet-associated collagenase inhibitor was found to be located in the alpha-granule. Studies with radiolabeled inhibitor failed to detect uptake of inhibitor by platelets. Furthermore, immunologically reactive protein of similar quantity to that found in platelets was identified in human megakaryocyte lysates. Thus, the data suggest that the collagenase inhibitor is endogenously produced and stored within platelet alpha-granules. The platelet derived collagenase inhibitor was antigenically identical to the collagenase inhibitor from human skin fibroblasts in double immunodiffusion and, like its fibroblast counterpart, inhibited collagenase on a 1:1 stoichiometric basis. When subjected to several of the chromatographic procedures utilized to purify the fibroblast protein, the platelet inhibitor behaved in an indistinguishable manner. Platelet factor 4, previously reported to be a collagenase inhibitor, was found to be immunologically unrelated to the platelet-derived collagenase inhibitor. Furthermore, platelet factor 4 displayed no collagenase inhibitory activity. Although the function of platelet-derived collagenase inhibitor is unknown, such a protein released by activated platelets may serve to regulate collagen turnover during the early stages of the inflammatory process. PMID- 2986138 TI - Evolution of mouse major histocompatibility complex genes borne by t chromosomes. AB - Virtually all wild mouse populations carry t haplotypes that cause embryonic lethality or semilethality, distortion of segregation ratios, suppression of crossing-over, and male sterility. The t complex of genes is located on chromosome 17, closely linked to the H-2, the major histocompatibility complex of the mouse. The t haplotypes differ from each other not only in lethal genes they carry but also in their linked H-2 haplotypes. In this study, we compared the class II H-2 genes present on 31 t chromosomes extracted from wild populations in different parts of the world. The comparison was based on the analysis of DNA fragments obtained after digestion with restriction endonucleases. The results reveal the existence of three major groups of class II alleles representing main branches on the evolutionary tree of the t chromosomes. Alleles within each group are similar if not identical, although they are borne by chromosomes that have been separated in time and space. The presence of similar alleles in Mus musculus and Mus domesticus suggests that some of them may have been separated for more than 1 million years. This must also be the minimal age of the t chromosomes but, because at least two of the three main branches appear to be related in their origin, the actual age of t chromosomes could be much greater. The observations support the proposal that H-2 genes evolve slowly. PMID- 2986139 TI - The telomeric region of the human X chromosome long arm: presence of a highly polymorphic DNA marker and analysis of recombination frequency. AB - A DNA fragment (named St14) derived from the human X chromosome reveals a small family of related sequences that have been mapped to the Xq26-Xq28 region by using a panel of rodent-human somatic cell hybrids. The probe detects in human DNA digested by Taq I a polymorphic system defined by a series of at least eight allelic fragments with a calculated heterozygosity in females of 80%. With Msp I, we found three additional restriction fragment length polymorphisms, each of them being defined by two alleles. These polymorphisms are also common in Caucasian populations. The genetic locus defined by probe St14 has been localized more precisely to the distal end of the X chromosome (in band q28) by linkage analysis to other polymorphic DNA markers. The results obtained suggest that the frequency of recombination is distributed very unevenly in the q27-qter region of the X chromosome, with a cluster of seven tightly linked loci in q28 showing about 30% recombination with the gene for coagulation factor IX located in the neighboring q27 band. Probe St14 reveals one of the most polymorphic loci known to date in the human genome, and 17 different genotypes have already been observed. It constitutes the best marker on the X chromosome and should be of great use for the genetic study of three important diseases: hemophilia A, mental retardation with a fragile X chromosome, and adrenoleukodystrophy. PMID- 2986140 TI - A drug-resistant mutation in the ribosomal DNA of Tetrahymena. AB - A mutation that confers resistance to the drug paromomycin is shown to be in the structural gene that codes for the ribosomal RNA in Tetrahymena. This observation was made by exploiting a variant of the ribosomal DNA that distorts amplification of this locus when a new somatic nucleus develops during conjugation. Because the allelic forms of this locus have a restriction endonuclease site polymorphism, it was possible to correlate drug resistance with presence of a specific allele. The genetic results have been confirmed by sequence analysis (presented elsewhere). Thus, the crosses presented here provide a unique opportunity to identify mutations in the ribosomal DNA. PMID- 2986141 TI - Insertion of long interspersed repeated elements at the Igh (immunoglobulin heavy chain) and Mlvi-2 (Moloney leukemia virus integration 2) loci of rats. AB - Restriction enzyme analysis of normal DNA derived from individual rats of the National Institutes of Health outbred Osborn-Mendel colony revealed that two independent single-copy loci, the Igh (immunoglobulin heavy chain) locus and the Mlvi-2 (Moloney leukemia virus integration 2) locus, a putative oncogene, are polymorphic (i.e., exhibit allelic variation). The polymorphism at both loci was due to the presence or absence of a long interspersed repeated DNA element (LINE). The LINE insertion in the Igh locus occurred in the joining (J) region, which is involved in the physiological rearrangement of this locus. The LINE insertion in the Mlvi-2 locus has occurred approximately 6 kilobases from the region of provirus integration in Moloney murine leukemia virus-induced rat thymomas. The two inserts are colinear with each other and with other randomly selected cloned copies of the rat LINE family, the general characteristics of which we also present. LINE insertion in the Mlvi-2 locus was observed in several rat strains, established from independent rat colonies, suggesting that LINE containing Mlvi-2 alleles may be widespread in the rat population. LINE insertion in the Igh locus was observed in 1 of 27 rats. The detection of a LINE-related polymorphism at two nonselected loci indicates that LINEs are transposable. The presence or absence of these long (greater than 5 kilobases), highly transcribed elements at single-copy loci could have profound effects on gene activity. Furthermore, LINE-containing single-copy loci could be affected by homologous interaction between the resident LINE and any of the other 50,000 or so copies of these elements in the rat genome. PMID- 2986142 TI - Restriction fragment length polymorphism of the human c-fms gene. AB - By using blot hybridization with a v-fms probe, a polymorphism for EcoRI, HindIII, and BamHI restriction endonuclease sites associated with the human c-fms locus was observed in a random adult population. This restriction fragment length polymorphism can be explained on the basis of the existence of two alleles, a and b, and is due to a short (congruent to 500 base pairs) deletion characteristic of allele a. The distribution in the analyzed population (48 unrelated individuals) is 23% heterozygotes ab, 75% homozygotes bb, and 2% homozygotes aa. Though the inheritance of this polymorphism follows a Mendelian pattern, the children from couples ab X bb are of the following genotype: 74% ab and 26% bb. These deviations from the expected frequencies of 50% suggest a selective pressure in favor of heterozygotes. PMID- 2986143 TI - Insertion of an unstable element in an intervening sequence of maize Adh1 affects transcription but not processing. AB - Three independently isolated and unstable mutants of the maize alcohol dehydrogenase 1 gene (Adh1) have arisen by insertion of the Mu transposing element into the first intervening sequence of the progenitor Adh1 allele. The mutants have been selected for their decreased levels of alcohol dehydrogenase 1 activity. The original mutants were unstable, giving rise to both revertant alleles and altered mutant alleles. From one of the original mutants, two derivative mutants have been recovered and described. We analyzed the effect of Mu insertion in all five of these mutants by measuring relative levels of run-off transcripts from the progenitor and mutant alleles and by comparing levels of run off transcripts corresponding to regions lying 5' and 3' to the insertion sites. In this paper we present evidence that early transcriptional events are affected, but that, in spite of the inclusion of a 1.4-kilobase transposing DNA element, processing of transcripts occurs normally. PMID- 2986144 TI - Nucleotide sequence preservation of human mitochondrial DNA. AB - Recombinant DNA techniques have been used to quantitate the amount of nucleotide sequence divergence in the mitochondrial DNA population of individual normal humans. Mitochondrial DNA was isolated from the peripheral blood lymphocytes of five normal humans and cloned in M13 mp11; 49 kilobases of nucleotide sequence information was obtained from 248 independently isolated clones from the five normal donors. Both between- and within-individual differences were identified. Between-individual differences were identified in approximately 1/200 nucleotides. In contrast, only one within-individual difference was identified in 49 kilobases of nucleotide sequence information. This high degree of mitochondrial nucleotide sequence homogeneity in human somatic cells is in marked contrast to the rapid evolutionary divergence of human mitochondrial DNA and suggests the existence of mechanisms for the concerted preservation of mammalian mitochondrial DNA sequences in single organisms. PMID- 2986145 TI - Predicted protein sequence of the murine I-E-beta S-polypeptide chain from cDNA and genomic clones. AB - Ia antigens are polymorphic cell-surface glycoprotein complexes, encoded within the I region of the mouse major histocompatibility complex, that control the ability of the organism to mount effective antigen-specific immune responses. We have isolated and determined the nucleotide sequences of cDNA and genomic clones for the I-E beta s gene and we present the predicted protein sequence for most of the E beta s polypeptide chain. The E beta s polypeptide shows 95% protein homology to the other cloned E beta alleles. Comparison of the protein sequences of five E beta alleles from haplotypes that differ in responder phenotype to pigeon cytochrome c suggests that the structure of one of the E beta hypervariable regions may determine responsiveness to this antigen. PMID- 2986146 TI - Cloning and expression of the Rickettsia prowazekii ADP/ATP translocator in Escherichia coli. AB - Cosmid clone banks of Rickettsia prowazekii genomic DNA were established in Escherichia coli and screened for expression of the rickettsial carrier-mediated ADP/ATP translocator. Out of 2700 clones screened, a single clone, designated MOB286, accumulated radioactivity when incubated with [alpha-32P]ATP in 100 mM sodium phosphate buffer. This clone carried a plasmid, pMW286, containing a 9 kilobase-pair insert of rickettsial DNA, as established by DNA X DNA hybridizations. Transformation studies with purified pMW286 established that the ability of E. coli cells to accumulate radioactivity was mediated by the recombinant plasmid. Results from experiments in which [3H]ATP was substituted for [alpha-32P]ATP strongly suggested that the radiolabeled ATP was transported intact. Furthermore, [3H]ATP was incorporated into 10% (wt/vol) trichloroacetic acid-precipitable material in a time-dependent manner. Uptake of ATP was also temperature-dependent, insensitive to atractyloside, N-ethylmaleimide, and dinitrophenol, and specific for ADP and ATP. Efflux of radiolabeled nucleotide was observed in the presence of extracellular ADP or ATP but not AMP and was not observed in the absence of extracellular adenine nucleotides. The successful cloning and expression of the rickettsial ADP/ATP translocator in E. coli will permit better characterization of rickettsial bioenergetics and of the metabolic regulation of obligate intracellular parasitism. PMID- 2986147 TI - Isolation and characterization of an olfactory receptor protein for odorant pyrazines. AB - The highly potent bell pepper odorant 2-isobutyl-3-[3H]methoxypyrazine [( 3H]IBMP) binds specifically and saturably to bovine and rat nasal epithelium. Specific binding is not detected in 11 other tissues assayed, and in the rat binding is 9 times higher in olfactory than in respiratory epithelium. We have purified to apparent homogeneity a soluble pyrazine odorant binding protein that constitutes approximately equal to 1% of the total soluble protein in bovine nasal epithelium. Polyacrylamide gel electrophoresis shows a single band of 19,000 Da and gel filtration data suggest that the native protein is a dimer of 38,000 Da. Binding of [3H]IBMP to the purified protein reveals two binding sites (Kd = 10 X 10(-9) M, Bmax = 135 pmol per mg of protein; Kd = 3 X 10(-6) M, Bmax = 25 nmol per mg of protein). The binding affinities of a homologous series of pyrazine odorants correlate with the human odor detection thresholds of these compounds. This correlation, together with the regional distribution of the protein, suggests that the protein is a physiologically relevant olfactory receptor. PMID- 2986148 TI - A mechanism for memory storage insensitive to molecular turnover: a bistable autophosphorylating kinase. AB - A mechanism is proposed for a molecular switch that can store information indefinitely, despite the complete turnover of the molecules that make up the switch. The design of the switch is based on known types of biochemical reactions. Central to the mechanism is a kinase that is activated by phosphorylation and capable of intermolecular autophosphorylation. It is shown that such a kinase and an associated phosphatase form a bistable chemical switch that can be turned on by an external stimulus and that is not reset by protein turnover. PMID- 2986149 TI - Increased glucagon receptors in chronically hypersomatotrophic and hyperglucagonemic rats. AB - The effect of increased levels of growth hormone on glucagon binding by isolated hepatocytes and on the cellular cyclic AMP response to glucagon was evaluated in rats bearing growth hormone-secreting tumor (Mt-T-W15) and in rats treated with rat growth hormone. An increased binding, due to an increased number of receptors, was observed in both groups of animals. Glucagon binding did not correlate with plasma glucagon levels, suggesting a failure of down regulation, possibly due to an effect of growth hormone and insulin on the number of receptors. Tumor-bearing and growth hormone-treated rats had larger hepatocytes so that, when hormone binding was expressed in terms of square micrometer of membrane surface, it appeared decreased. When the tumor was removed the increase in the number of glucagon receptors per cell persisted, even though the average cell size returned toward normal. It is suggested that this retention of the receptors may have been the result of continuing hyperinsulinism. Basal cAMP levels were elevated in hepatocytes of tumor-bearing and growth hormone-treated animals, possibly due to cell hypertrophy. On the other hand, the maximum cAMP response to glucagon was not altered by the experimental procedures. A negative effect of insulin on cAMP accumulation may explain this apparent paradox. Indeed, hepatocytes isolated from rats following tumor removal, but with continuing hyperinsulinemia, had a lower maximum cAMP response, even though the glucagon binding per cell or per unit of cell surface was increased. PMID- 2986150 TI - Herpes simplex virus latency in the rabbit trigeminal ganglia: ganglionic superinfection. AB - It has been confirmed and further documented that infection of the rabbit cornea with the E-43 strain of HSV-1 precludes superinfection of the corresponding trigeminal ganglia by another HSV strain, i.e., the challenging virus does not establish latency and can not be recovered from the ganglia. It was shown that after primary infection, a state of resistance is established in the neuronal cells of the ganglia, and although the challenging strain reaches the ganglia, it does not cause discernible acute infection, and does not displace the resident virus in the ganglia. This protection was present 6 months after primary infection, was independent of immune factors such as circulating or secretory antibodies, and was localized to the point of entry of the primary infecting strain and the sensory neurons that innervate that site. The smallest inoculum that provided protection from ganglionic superinfection was that which produced overt disease in the eye, although different degrees of disease resulted from varying inocula above this minimum. Asymptomatic primary infections produced by subminimal inocula of the E-43 strain or by the HSV recombinant strain, F(MP)F, which is avirulent for the rabbit eye, protected against severe disease and death, but the degree of protection against ganglionic superinfection was variable and depended on the time of challenge. These findings suggest that susceptible neurons in the trigeminal ganglion, when "occupied" by an infecting strain, cannot be superinfected by a second strain. PMID- 2986152 TI - Anthrax toxin components stimulate chemotaxis of human polymorphonuclear neutrophils. AB - Effects of the three-component toxin of Bacillus anthracis on chemotaxis of human polymorphonuclear leukocytes (PMN) were investigated in an effort to determine the basis of the reported antiphagocytic effect of the toxin. The three toxin components, edema factor (EF), protective antigen (PA), and lethal factor (LF), were tested alone and in various combinations for their effect on PMN chemotaxis under agarose to formyl peptides and zymosan-activated serum. No component was active alone; combinations of EF + PA, LF + PA, and EF + LF + PA markedly stimulated chemotaxis (directed migration), but had little or no effect on unstimulated random migration. The toxin components were not themselves chemoattractants. EF in combination with PA had previously been identified as an adenylate cyclase in Chinese hamster ovary (CHO) cells. We found that EF + PA produced detectable cyclic adenosine 3'-5'monophosphate (cAMP) in PMN, but the level of cAMP was less than 1% of that produced in CHO cells by EF + PA, and in PMN by other bacterial adenylate cyclases. LF + PA (which stimulated chemotaxis to an equivalent extent) had no effect on cAMP levels. Thus, the enhancement of chemotaxis by anthrax toxin (at least by LF + PA) does not seem to be related to adenylate cyclase activity. PMID- 2986151 TI - Insulin-like growth factor II binding to cultured human chondrosarcoma cells. AB - Cultured cells originally derived from a human chondrosarcoma (A1684) were used to investigate somatomedin binding in terms of kinetics and specificity. In this study, the rat somatomedin, multiplication-stimulation activity (MSA) was utilized. While the human chondrosarcoma cells did not exhibit a mitogenic response to MSA, the rate of transport of glucose and amino acids was significantly increased. In competitive binding experiments a specific insulin insensitive MSA receptor was identified which showed half maximal displacement of tracer at a concentration of 250 ng/ml of MSA using whole cells. This receptor had an affinity constant of 4.8 X 10(7) M-1. Kinetic analysis of MSA binding to membrane preparations and to Triton X-100 solubilized membranes revealed an increase in the binding affinity to 1.28 X 10(8) M-1 and 2.8 X 10(8) M-1, respectively. Of particular significance is the observation that these cells have especially high levels of MSA receptors. Determination of binding capacity revealed that these cells contain approximately 1.9 X 10(6) MSA receptors per cell and therefore are an excellent model system for the characterization and purification of somatomedin receptors. Affinity labeling of the MSA receptor using the chemical crosslinking reagent, disuccinimidyl suberate, confirmed that this receptor was of the type II class of somatomedin receptors and exhibited a molecular weight of 218,000 under nonreducing conditions. PMID- 2986153 TI - Systemic and renal hemodynamic responses to vascular blockade of vasopressin in conscious dogs with ascites. AB - A role for arginine vasopressin has been implicated in the compensatory control of arterial blood pressure in several animal models with reported increases in plasma levels of arginine vasopressin. A threefold elevation in plasma vasopressin has been reported in conscious dogs following constriction of the inferior vena cava. In the present study, infusion of the arginine vasopressin antagonist [1-(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid), 2-O methyltyrosine] Arg8-vasopressin into conscious dogs with chronic caval constriction did not decrease mean arterial blood pressure. However, the dose of infused antagonist completely blocked the pressor response to 2 micrograms of exogenous vasopressin. Also the antagonist produced no effect on heart rate, plasma renin activity, or urinary volume and electrolyte excretions. A slight, transient increase (P less than or equal to 0.05) was observed in creatinine clearance and in PAH clearance following antagonist infusion, suggesting a possible decrease in renal vascular resistance. These data suggest that the direct vasoconstrictor actions of vasopressin contribute minimally, if at all, to blood pressure maintenance following chronic caval constriction. Alternatively, blockade of endogenous vasopressin receptors at the level of peripheral arterioles may have resulted in no depressor response due to a masking of this response by other compensatory hormonal and neural pressor systems. PMID- 2986154 TI - Temporal relationships of treatment with angiotensin converting enzyme inhibitors on the hypertension and cardiac hypertrophy in spontaneously hypertensive rats. AB - Two groups of spontaneously hypertensive rats (SHRs) were treated with captopril and enalapril, respectively. Treatment was started at weaning and at 8 months of age. Blood pressure was maintained close to normal in rats treated from weaning and significantly lowered but not to normal levels in rats started on treatment at 8 months. The long-term treated rats did not show significant cardiac enlargement whereas those treated at 8 months had an increase in cardiac size although not to the same level as untreated animals. Treatment with minoxidil and minoxidil plus propranolol similarly lowers blood pressure, but does not reduce myocardial hypertrophy. The data confirm that both the time of onset of treatment as well as the type of drug influence the myocardial complications of hypertension in SHRs. These studies further demonstrate the use of the SHR as a model for clinical pharmacologic studies with antihypertensive agents. PMID- 2986155 TI - Complement-mediated arachidonate metabolism. AB - The leukotrienes are important mediators of numerous responses in lung tissue. Both direct injury and immune injury result in the production of these arachidonate products. Several cellular components participate in the immune surveillance system including monocytes, mast cells and PMNs. Each cell type produces different quantities and types of leukotrienes in response to ionophore (A23187) activation. A common feature shared by each of these cells is control of arachidonic acid metabolism at the level of the 5-lipoxygenase. One provocative interpretation of our results is that the 5-lipoxygenase is activated by C5a and that concomitant modulation of 5-lipoxygenase activity provides a means whereby arachidonic acid metabolism is directed in these cells to either the cyclooxygenase or lipoxygenase pathway. Another common feature that these cells share is that they utilize arachidonic acid mobilized from other cells such as stimulated platelets, certain monocytes, or even damaged tissue. For example, free nonesterified arachidonic acid has been measured at 100 microM in inflamed tissue. Therefore, fluctuations in exogenous arachidonate levels may provide a significant modulation of the inflammatory response by controlling the levels of lipoxygenase products formed by leukocytes. In this scenario the humoral factor C5a is the initiator of the host's response to provide a variety of functional arachidonate products. Another feature that the cellular components of the immune system share is that they may utilize other exogenous lipid substrates. In this case, a lipid product of one cell type may serve as a signal or substrate for a second cell's lipoxygenase pathway. This hypothesis may explain the apparent synergy observed in this and other studies when mixed cell populations were activated. Several hydroperoxy lipids are proposed to be regulatory for the lipoxygenase pathway. Another valid interpretation could be that 5-hyperoxy eicosatetraenoic acid and LTA4 produced in one cell may diffuse to another cell and be utilized by the lipoxygenase pathway of that cell type. From the results of this study we conclude that the secondary mediator profile obtained when cells are activated by arachidonic acid and C5a depends on the cell composition. We can extend this interpretation of our results to explain two seemingly opposite results obtained when C5a is administered to experimental animals either intrabronchially or intravenously. Future evaluations of the biological effects of C5a should therefore take into consideration the composition of the cells at the target tissue site.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986156 TI - Airway inflammation and asthma. Importance of arachidonate metabolites for airway hyperresponsiveness. PMID- 2986158 TI - Measurement of 5-lipoxygenase products in the lung. PMID- 2986157 TI - Beta-adrenoceptor desensitization in the lung. A phenomenon related to prostanoids. PMID- 2986159 TI - Pathogenetic correlations among different types of digital malformations. PMID- 2986160 TI - Approaches used for the detection of infectious agents as human teratogens. AB - A review of the events which lead up to the recognition of six infectious agents as teratogens showed that in most cases the associations were made through studies of the pathology of the disease or in association with epidemics which affected large numbers of pregnant women. Epidemiological studies of various populations were then conducted to obtain more complete information about the frequency of infection; the syndrome produced in the child; the effect of time of maternal infection; the method of transmission of infection; and approaches to diagnosis, treatment, and prevention of the disease. Investigation of other possible teratogens should emphasize examination of the pathology of the disease and the analysis of pregnancy outcomes of large numbers of women who experience a given infection. PMID- 2986162 TI - Prostaglandins, catecholamines, and differentiation of the secondary palate. PMID- 2986164 TI - Reduction limb defects in Finland. PMID- 2986163 TI - The role of the central noradrenaline system in regulating neuronal plasticity in the developing neocortex. PMID- 2986161 TI - Neurotoxicity of opiates during brain development: in vivo and in vitro studies. PMID- 2986165 TI - DNA repair in the in vivo site specific excision of palindromic sequences. PMID- 2986166 TI - Carcinogen-induced insertion mutations in E. coli. AB - Mutagenic effects of AAF have been examined by transforming competent E. coli cells with plasmid DNA which were regionally-modified with this carcinogen. Transposition of IS5 from chromosomal DNA to target sequences in the AAF-modified region was detected in six plasmids obtained from wild type and recA strains. A single GC addition was identified in an additional plasmid; no deletions were detected. The observed frequency of IS5 insertion, 2.5 X 10(-3), exceeds that reported for naturally-occurring IS5 transposition in E.coli. These results suggest that the frequency of recombinational events in E.coli can be increased by the presence of a DNA-carcinogen adduct. PMID- 2986167 TI - Some electrostatic aspects of the interaction between DNA or lipids and WR-2721 or its metabolite WR-1065. PMID- 2986168 TI - Netropsin binding to polyd(AT) X polyd(AT) and to polydA X polydT: a comparative thermodynamic study. PMID- 2986169 TI - Peptide-nucleic acid interactions: possible recognition determinants of Eco RI endonuclease. PMID- 2986170 TI - Modulation of cell surface antigens accompanies morphological differentiation of human neuroblastoma cell lines. PMID- 2986171 TI - The expression of tubulin and various enzyme activities during neuroblastoma differentiation. PMID- 2986172 TI - Expression of a melanocyte phenotype in human neuroblastoma cells in vitro. PMID- 2986173 TI - Clinical studies with neuron specific enolase. PMID- 2986174 TI - NSE in neuroblastoma and other round cell tumors of childhood. PMID- 2986175 TI - Neuronal, Schwannian and melanocytic differentiation of human neuroblastoma cells in vitro. PMID- 2986176 TI - Tumor differentiation--application of prostaglandins in the treatment of neuroblastoma. PMID- 2986177 TI - Gamma-linolenic acid inhibits the development of the ethanol-induced fatty liver. AB - In the context of recent work showing numerous interactions between ethanol, essential fatty acids (EFA) and prostanoids, we have evaluated the effects of gamma-linolenic acid methyl ester (GLA 99%; 18:3, n-6), on hepatic pathology induced by ethanol in rats. Groups of animals were pair-fed an alcohol-containing liquid diet or an iso-caloric maltose-dextrin diet. Animals fed ethanol for ten days had markedly increased hepatic triglycerides and histological evidence of fatty liver. These effects were partially attenuated by administration of GLA during the period of ethanol administration. PMID- 2986178 TI - Hypothesis: on the similar prostaglandin mediation in the anti-hypertensive action of alpha- and beta- adrenoceptor antagonists; confusion of present system of receptor classification? PMID- 2986179 TI - Interactive effects of caffeine, 2-chloroadenosine and haloperidol on activity, social investigation and play fighting of juvenile rats. AB - The effects of caffeine, 2-chloroadenosine and haloperidol and their interaction on activity, social investigation, and two measures of play fighting (crossover and pinning), were investigated in juvenile male rats. Caffeine (20 mg/kg) increased activity, decreased crossover and pinning, but had no effect on social investigation. Both 2-chloroadenosine (0-10 mg/kg) and haloperidol (0-10 mg/kg) depressed activity, social investigation, crossover and pinning. When given together in varying dosages, caffeine and 2-chloroadenosine had behavioral effects suggestive of a competitive interaction between the two drugs. In contrast, the effects of haloperidol were not appreciably altered by simultaneous caffeine treatment. These results suggest that the influence of caffeine and 2 chloroadenosine on activity, social investigation and play fighting involve interaction with adenosine receptors. PMID- 2986180 TI - Morphine and naloxone effects on tonic immobility and the dorsal immobility response in the rat. AB - Adult male Wistar rats treated with morphine sulfate (0.5, 5.0, and 10.0 mg/kg, SC) showed a dose-dependent potentiation of tonic immobility (TI) and dorsal immobility response (DIR) durations. Naloxone (4.0 mg/kg, SC) did not affect the DIR durations but reversed the potentiated morphine effects on both TI and DIR. These results suggest that although opiate receptors may be involved, mu-opiate receptor function is not essential for modulating various complex immobility responses. PMID- 2986181 TI - Effects of chronic LiCl and RbCl on muricide induced by midbrain raphe lesions in rats. AB - Midbrain raphe lesions in rats (raphe rats) induce aggressive behavior including muricide. A single administration of LiCl (Li) 100 mg/kg to raphe rats produced only 25% of muricide inhibition. However, the inhibitory effect of muricide in raphe rats significantly increased from the 5th day following repeated administration of Li. Chronic Li also inhibited muricide in olfactory bulbectomized (OB) rats. The inhibition of muricide lasted until the next day to some extent. In this point, the effect of Li on muricide is similar to that of antidepressants, but not of neuroleptics. On the contrary, RbCl (Rb) showed a tendency to induce muricide. The single re-administration of Li 100 mg/kg on the withdrawal on the 7th day after repeated administration for 14 days showed a significant inhibition of muricide in raphe rats, unlike that in OB rats. Li also showed a partial prophylactic effect on muricide when Li 100 mg/kg was administered for 1 week before raphe lesions. These results suggest that raphe rats may serve not only as an experimental model of depression, but also as that of manic illness. PMID- 2986182 TI - Habituation to repeated stress is stressor specific. AB - Rats were exposed to 15 min of restraint or footshock or forced running in an activity wheel once a day for 10 days. Control groups were handled only. On the 11th day, rats from each stressor group and controls were exposed to 15 min of one stressor in a crossed design such that all combinations of one chronic stressor and one acute stressor were performed. Rats were sacrificed immediately following removal from their home cage or after 15 min stressor exposure on the 11th day and plasma corticosterone and prolactin and pituitary cyclic AMP levels were determined. There were no measured differences in these stress indices among groups of rats sacrificed immediately upon removal from their home cage on day 11 regardless of previous history on days 1 through 10. Plasma corticosterone and plasma prolactin and pituitary cyclic AMP levels were elevated in all rats exposed to any of the three stressors immediately prior to sacrifice as compared to all rats not exposed to stress immediately before sacrifice. However, plasma prolactin and pituitary cyclic AMP responses to each of the 3 stressors were attenuated in rats which had previous exposure to that specific stressor as compared to rats which had previous experience with a different or no stressor. We conclude that habituation results from behavioral experience with a particular stressor rather than biochemical adaptation resulting from repeated challenge to hormonal and neurochemical systems responsive to stress. PMID- 2986183 TI - Intracerebroventricular saline treatment elevates the pain threshold. Is this phenomenon mediated by peripheral opiate receptors? AB - Intracerebroventricular (i.c.v.) saline treatment of unanaesthetized mice resulted in moderate but well detectable analgesia. This elevation of pain threshold (by 60-90%) could be observed already 2 min after i.c.v. injection and lasted 15-30 min. Surprisingly, the analgesic action of i.c.v. treatment was not attenuated if naloxone was administered i.c.v. instead of saline. In fact if applying it in higher doses, the opiate antagonist potentiated the antinociceptive action. However, giving naloxone subcutaneously (s.c.) before i.c.v. saline treatment, the resulting analgesia was significantly reduced. It is tentatively put forward that i.c.v. treatment is stressful and in itself may induce mobilization of endogenous opioids and the latters elevate the pain threshold via peripheral opiate mechanisms. PMID- 2986184 TI - [The stability of nourseothricine in bentonite adsorbate and aqueous solutions]. PMID- 2986185 TI - Effects of ethylketocyclazocine on adrenergic transmission in the isolated perfused cat spleen. AB - The pre- and postsynaptic effects of ethylketocyclazocine (EKC), a putative kappa opiate receptor agonist, on sympathetic adrenergic neurotransmission were investigated using the isolated perfused cat spleen. Perfusion of spleens with EKC (10(-7) and 10(-6)M) produced a modest dose-dependent inhibition of nerve stimulation-mediated overflow of norepinephrine (NE) and total 3H. In contrast, EKC (10(-4)M) produced an increase in the release of NE, and total 3H overflow. Perfusion with EKC (10(-5) and 10(-4)M) resulted in a dose-dependent inhibition of the postsynaptic response to stimulation-mediated release of NE. Likewise, EKC inhibited splenic contraction due to exogenously administered NE which was not antagonized by naloxone. These results suggest that EKC exerts a direct effect on peripheral adrenergic neurotransmission not mediated via classical kappa opiate receptors. PMID- 2986186 TI - Nucleotide sequence analysis of a gene encoding a streptomycin/spectinomycin adenylyltransferase. AB - The nucleotide sequence of 1400 bp from R-plasmid R538-1 containing the streptomycin/spectinomycin adenyltransferase gene (aadA) was determined, and the location of the aadA gene was identified by a combination of insertion and deletion mutants. Its gene product, aminoglycoside 3"-adenylyltransferase (AAD(3")(9), has a Mr of 31,600. PMID- 2986187 TI - The integrated and free states of Streptomyces griseus plasmid pSG1. AB - A 16.6-kb plasmid-pSG1-was isolated from Streptomyces griseus following transformation of protoplasts with unrelated plasmids. Southern hybridization experiments with radioactive probes prepared from pSG1 fragments and immobilized S. griseus DNA fragments indicated that the plasmid was present in the progenitor strain, in an integrated state. In the pSG1+ isolates plasmid sequences existed both as integrated sequences and as free plasmids. The integrated state of maintenance persisted in strains which have been cured of the free plasmid. The junction site on the plasmid was located on a 0.5-kb EcoRI-SalI fragment. The chromosomal integration site was demonstrated to be the same in all strains derived from S. griseus NRRL3851. The occurrence of both states of plasmid maintenance in the same clones indicates that an integrated pSG1 sequence does not interfere with free plasmid replication and partition. It suggests that the establishment of the free state may involve a replicative excision of pSG1 from the S. griseus chromosome. PMID- 2986188 TI - A two-stage molecular model for control of mini-F replication. AB - It is known that mini-F replication requires production of a 29,000-Da protein, protein E, and origin of replication sequences mapping around 45. kb. Further, control of replication is determined by two genes, copA and copB. In the present work a description is given of the cloning of an F restriction fragment containing the amino terminal portion of the protein E gene, repE, and associate promoter activity. It is shown that expression of this promoter is negatively regulated in trans by sequences taken from the F replication region of copA+ plasmids. However, the same sequences taken from six different copA- plasmids failed to repress expression of the promoter. Since prior studies have shown that copA+ determines a repressor of replication, it is now suggested that the above results are an accounting of where this repressor works. A hypothesis is also proposed to explain control of F replication by the copA and copB regulatory genes. PMID- 2986189 TI - A small mobilizable IncP group plasmid vector packageable into bacteriophage lambda capsids in vitro. AB - A mobilizable cosmid derivative of an IncP group plasmid was constructed by cloning the oriT region of RK2, a wide host-range plasmid, and the minimal DNA sequence of bacteriophage lambda required for efficient packaging in vitro. This cosmid is 13 kb in size and has unique restriction sites for EcoRI, XhoI, HindIII, and SalI. The XhoI and HindIII sites are within the kanamycin-resistance gene and the SalI site is in the tetracycline-resistance gene. This plasmid was mobilizable from an Escherichia coli donor to a number of diverse gram-negative bacteria at a frequency of 0.8 to 10 per 100 donors. This vector is one of the smallest of all wide host-range cosmids described in the literature. As part of this study, another mobilizable IncP group plasmid vector has also been constructed which, in addition to the sites listed above, has a unique BglII site, but which lacks the packager sequence. PMID- 2986190 TI - Restriction endonuclease mapping of R27 (TP117), an incompatibility group HI subgroup 1 plasmid from Salmonella typhimurium. AB - A circular map of the IncHI plasmid R27 corresponding to a genome size of 182 kb was established using the restriction endonucleases ApaI, XbaI, and PstI. The map was derived from the results obtained by hybridizing individual ApaI and XbaI fragments to blotted digests of the plasmid, as well as from complete and partial digests. Analysis of a deletion mutant derived by in vitro digestion with PstI and of transfer-defective and tetracycline-sensitive deletion mutants of R27 derived by Tn5 insertion were instrumental in determining the positions of some fragments. PMID- 2986191 TI - Radioisotope techniques in the evaluation of patients with coronary artery disease. AB - Cardiac scintigraphy provides an exciting new diagnostic modality. Currently, its two major clinical applications are the identification of patients with ischemic heart disease and the characterization of ventricular function. These techniques also appear to have utility in assessing the size of myocardial infarction, the severity of obstructive coronary disease, and the effects of medical or surgical therapy. Continued careful correlation of clinical and scintigraphic findings will make these techniques even more useful in the future. PMID- 2986192 TI - Characterization of platelet alpha 2 adrenoceptors and measurement in control and depressed subjects. AB - The alpha-adrenoceptor on platelets has been characterized using 3H-yohimbine, 3H dihydroergocriptine, and 3H-clonidine. The receptor, which exhibits the characteristics of an alpha 2-type, has a Bmax for dihydroergocriptine of 330 fmoles/mg protein, for yohimbine of 178 fmoles/mg protein, and for clonidine of 38 fmoles/mg protein. Clonidine, but not yohimbine binding, is decreased by the presence of K+, Na+, or Li+. Adenosine triphosphate (ATP) and the guanosine triphosphate (GTP) analogue, Gpp(NH)p, reduce the affinity of clonidine for its binding site. Acute exercise, such as playing squash, does not apparently alter the Bmax or Kd of 3H-yohimbine binding to platelet membranes, and in vitro studies, with intact platelets or platelet membranes, show that incubation with adrenalin (10 microM) does not induce alterations in Bmax or Kd. In the present study, no correlation was found between age and alpha 2-adrenoceptor numbers. There was no significant difference in the Bmax for 3H-yohimbine binding to platelet membranes from control and depressed subjects, although the mean value for the depressed group was some 10% lower than that for the corresponding control group. There were no overall significant and consistent effects of desipramine (DMI) treatment. After 2-3 days of treatment, the Bmax was reduced by 20%, after 7 days by 14%, and after 21 days it was 8% above the control value. When an additional group of patients on DMI (7 days of treatment) was analyzed using one supramaximal concentration of 3H-yohimbine, there was a significant decrease (25%) in binding. PMID- 2986193 TI - [Extension alveoloplasty (I). New surgical procedures for the treatment of alveolar collapse and residual alveolar ridge atrophy]. PMID- 2986195 TI - Intra-arterial digital subtraction angiography. AB - DSA is an imaging technique that should be integrated into a state-of-the-art angiographic system for proper application and utilization. This results in improvements in both efficiency and diagnostic accuracy. However, from the physician's viewpoint, the proper use of a DSA imaging facility mandates not only an understanding of angiographic principles as applied with conventional film screen systems but also an in-depth understanding of the factors that affect DSA performance. In particular, factors affecting spatial resolution and contrast sensitivity are crucial. This knowledge has to be applied interactively and "on line" to achieve optimal IA-DSA image quality. PMID- 2986194 TI - [Extension alveoloplasty (II). New surgical procedures for the treatment of alveolar collapse and residual alveolar ridge atrophy]. PMID- 2986197 TI - [Ultrasonic diagnosis of the breast]. PMID- 2986196 TI - [Pharmacoarteriography of gestation-induced invasive trophoblastic tumor (ITT)]. PMID- 2986198 TI - Biological alterations in the primary affective disorders and other tricyclic responsive disorders. AB - Noradrenergic function was studied in patients with primary affective disorder and other tricyclic-responsive disorders including obsessive-compulsive disorder, anorexia nervosa and panic attack/agoraphobia in medication-free states. Pre synaptic noradrenergic activity was assessed by assaying plasma concentrations of norepinephrine (NE) and its metabolite 3-methoxy,4-hydroxyphenylglycol (MHPG). Noradrenergic receptor responsiveness was evaluated by measuring plasma growth hormone (GH), MHPG, and NE responses to clonidine. Binding of tritiated dihydroergocriptine (3H-DHE) and biochemical responsiveness of alpha 2-adrenergic receptors were measured in platelet preparations. These studies suggest that noradrenergic activity may be altered in several tricyclic-responsive disorders and are consistent with the possibility that tricyclic antidepressants may serve to stabilize a dysregulated noradrenergic system in patients from several diagnostic categories. PMID- 2986200 TI - [Sex pheromones and plasmid transfer in Streptococcus faecalis]. PMID- 2986199 TI - Therapeutic responses to tricyclic antidepressants and related drugs in non affective disorder patient populations. AB - Although therapeutic responsiveness to tricyclic antidepressants has been primarily associated with the affective disorders, clinical investigations in the last decade have suggested that non-affective disorders such as panic disorder, obsessive-compulsive disorder, anxiety disorder, bulimia, enuresis, migraine, and the chronic pain syndrome may also respond to tricyclics and other antidepressants. This therapeutic responsiveness may sometimes be related to improvement in secondary depressive symptoms, but may also clearly occur in the absence of secondary depression; in particular, improvement in the core symptoms of at least some of these disorders may occur without a change in mood. Furthermore, many patients with these disorders display psychobiologic abnormalities that show many similarities, but also some differences, compared to those observed in patients with affective disorders, despite the frequent absence of affective symptoms. While an improvement in subclinical or "masked" depression remains one hypothesis linking tricyclic responsiveness and shared biological abnormalities in this diverse group of diagnostic entities, an alternative hypothesis (the "ven disorder" hypothesis) is presented, suggesting the possibility that tricyclic and other antidepressant-responding patients have a core disorder with common psychobiologic abnormalities but multiple clinical and diagnostic presentations. An alternative hypothesis (the "shotgun" hypothesis) suggests that the multiple actions of tricyclics (e.g. on adrenergic receptors vs. muscarinic receptors vs. serotonin system changes) may each be differentially important in the therapeutic outcome in patients with specific or predominant problems in one or another of these areas. An examination of both the similarities and differences among the non-affective, tricyclic-responsive disorders and the affective disorders may provide clues about the important psychobiologic elements in these disorders, and to the mode of action of tricyclic antidepressants and related drugs across the psychiatric disorder spectrum. PMID- 2986201 TI - Surgical splint construction for hydroxylapatite augmentation of the inadequate mandibular residual ridge. PMID- 2986202 TI - Solubilization and characterization of CCK receptors from mouse pancreas. AB - To study the characteristics of the CCK receptor, plasma membranes were prepared from mouse pancreatic acini, and CCK receptors solubilized with 1% digitonin. To measure hormone binding, the solubilized receptors were incubated with 125I-CCK at 4 degrees C and the hormone-receptor complex was precipitated with 10% polyethylene glycol. Specific 125I-CCK binding by the solubilized CCK receptor was compared to that by the plasma membrane-bound CCK receptor. Both the solubilized and the membrane-bound receptor displayed optimal binding at an acidic pH (between 6.0 and 7.0) and showed a similar sensitivity to monovalent and divalent cations. The solubilized receptors preserved their relative specificity for CCK molecules: CCK-8 greater than CCK-33 greater than desulfated CCK-8 greater than CCK-4. However, the soluble CCK receptor had a lower binding affinity than plasma membrane-bound receptor. Solubilized receptors preserved their relative specificity for inhibitors of CCK binding and action: dibutyryl cyclic GMP greater than N-CBZ-tryptophan greater than proglumide. Solubilized receptors had affinities for these antagonists that were similar to receptors on intact plasma membranes. These data indicate, therefore, that the specific binding properties of the CCK receptor are inherent to the solubilized glycoprotein molecules. PMID- 2986203 TI - Up-regulation of thyrotropin-releasing hormone (TRH) receptors in rat spinal cord after codepletion of serotonin and TRH. AB - Immunohistochemical evidence indicates the coexistence of serotonin and TRH in many raphe neurons. We examined the biochemical changes in TRH receptors after destruction of the serotonergic pathways by 5,7-dihydroxytryptamine (5,7-DHT). 2 weeks after an intracerebroventricular injection of 5,7-DHT, rats were killed, and specific brain regions were dissected on ice. Serotonin levels in the CNS of lesioned rats was reduced by 50-85% in all regions, with the highest reduction in the spinal cord and hippocampus. Immunoreactive TRH was reduced in the spinal cord by 70%, but other brain regions contained normal levels of TRH. TRH receptor binding was increased by 40% in the spinal cord of lesioned rats, but appeared unchanged in rostral brain regions in which no decrease in TRH content was detected. Scatchard plots of TRH receptor binding in the spinal cord indicated that the increased binding after 5,7-DHT administration reflected an increased receptor number. These findings suggest that biochemical up-regulation of TRH receptors occurs in the spinal cord following depletion of TRH. PMID- 2986204 TI - Imaging neuroreceptors in the human brain in health and disease. PMID- 2986206 TI - [Prolactin and estradiol levels in the plasma of patients with hepatic cirrhosis]. PMID- 2986205 TI - [Receptors for polymerized human albumin: new aspects of hepatitis B virus infection]. PMID- 2986207 TI - [Neurological manifestations in systemic lupus erythematosus. Analysis of 22 patients]. PMID- 2986208 TI - [Deficit of ACTH manifested as hyponatremia]. PMID- 2986209 TI - [Computed tomographic follow-up of brain volume fluctuations before and after alcohol withdrawal treatment]. AB - CT follow-up studies were done in 65 alcoholics before an inpatient treatment and after a period with confirmed abstinence of 5 weeks duration. The scans were rated "blind" by linear measurement of well defined distances. An improvement (significant reduction of brain "atrophy") was found in 33 patients (50.8%), 5 patients (7,7%) showed a trend towards progression of brain "atrophy". The possibility of recovery tends to be significantly greater in younger subjects. These findings and the results of recent MR follow-up studies are consistent with decreased free water during alcohol intoxication and an increase in brain water during alcohol withdrawal. PMID- 2986210 TI - [CT findings in chordomas of the base of the skull]. AB - Nine chordomas of the skull base and their CT appearances are described. Destruction of the clivus and of the parasellar portions of the middle fossa are characteristic, as is calcification which may be spotty, coarse or ill-defined. The tumour is characterised by marked enhancement following contrast but, on the unenhanced CT, contrary to statements in the literature, it is hypodense or isodense with respect to cerebral tissue. The tendency to recurrence and an altogether poor prognosis is confirmed. PMID- 2986211 TI - [Splenic infarct in the computed tomogram]. AB - Splenic infarcts are represented by wedge-shaped, oval or linear areas. Haemorrhagic infarcts are characterised by being hyperdense. Disseminated infarction occurs predominantly in myeloproliferative diseases. During the early stages, the infarct appears as an ill-defined hypodense defect, with non homogeneous contrast enhancement. During the acute and sub-acute stage, the density of the infarct is low and there is no contrast enhancement. During the chronic stage, its density increases and there is slight contrast enhancement. Complications following splenic infarcts, such as abscesses, bleeding and rupture can be demonstrated by CT with great accuracy. Problems in differential diagnosis may occur if there are atypical manifestations of the infarct, with respect to abscess or leukaemic infiltrations. PMID- 2986212 TI - [Diagnosis of choledochal cysts. The methodological value of imaging procedures]. AB - For 10 cases in which we detected cysts in the choledochus ourselves comparing traditional radiological methods (infusion-cholegram, ERC, scintigraphy, barium meal examination, angiography) with recent imaging procedures (ultrasound, CT, NMR) the following sequence of procedures proved to be favorable: Screening methods are ultrasound and infusion-cholegram. CT and NMR furnish good presentations of the intra- and extrahepatic dilatations of the bile duct. ERC still represents the best methods for demonstration of an extrahepatic cyst of the choledochus. Hepato-biliary functional scintigraphy is performed as a supplement. Barium meal examination and coeliacography furnish a small diagnostic contribution only. PMID- 2986213 TI - Primary malignant tumours of the duodenum. AB - The clinical and radiological findings in 19 patients with primary duodenal malignancy are described. Weight loss, abdominal pain, nausea and vomiting were the main symptoms. Diagnosis was made by endoscopy or ERCP (71%) or by barium studies (68%). In retrospect the tumour was visible in 97% of the studies. Tumour growth was longitudinal, circular or spiral, the inner curvature being involved over a greater length than the outer curvature. Exophytic tumour growth, involvement of the papilla of Vater, malignant spikes, transient, non-constant tumour image, skip lesions and ulceration were often seen. Mean survival time was 18 months from start of symptoms in 10 inoperable patients, and 24 months in 9 patients undergoing resection. PMID- 2986214 TI - [Computed tomographic diagnosis and differential diagnosis of juvenile angiofibroma and angiomatous polyps]. AB - Juvenile angiofibromas (JAF) and angiomatous polyps (AP) are vascular histologically benign tumors presenting with an identical clinical triad. Both lesions can be differentiated by their typical geographic pattern of growth, different pattern of tumor extension and their highly characteristic angiographic and angiocomputertomographic appearance. Time-density curves were of no use in differentiating the big variety of hypervascular lesions from each other. CT images of the vascular tumor phase after bolus injection of contrast medium however showed interesting pathomorphological details, which were extremely useful in differentiating JAF from AP and other vascular tumors. It is our opinion, that invasive angiography for diagnosing JAF has been surpassed, but is further essential for mapping of tumor perfusion, feeding vessels and praeoperative embolization therapy. PMID- 2986215 TI - [A new 3-dimensional imaging method in computed tomography]. AB - A new imaging method for computerized tomography is presented and some examples are demonstrated. The method is based on the collection of all image data of a chosen block of tissue and enables the examiner to perform multiplanar reconstructions of high quality in any direction, even in oblique planes. In order to achieve a better orientation, the block of tissue can also be visualized at the same time on the reconstruction. PMID- 2986216 TI - [Estimation of heart size--x-ray versus 2-dimensional echocardiography]. AB - Radiologic heart volume measurement (HV) and two dimensional echocardiographic estimation of left ventricular volume and ejection fraction (EF) were carried out in 108 healthy subjects and 68 patients with hypertony, dilatative cardiomyopathy and hypertrophic-obstructive cardiomyopathy (HOCM). Healthy subjects show a close relationship between echocardiographic determined volume and absolute heart volume (HV), in patients with hypertony, dilatative cardiomyopathy and HOCM there is also a significant correlation. However, no correlation exists between EF and HV and HV/kg in patients with hypertony and HOCM, whereas patients with dilatative cardiomyopathy show a high significant inverse relationship. It is concluded that these results are caused by great differences in muscle mass. The clinical value of radiologic heart volume as a parameter of left ventricular systolic function seems to be low and depends on cause of the heart enlargement. PMID- 2986217 TI - [Comparison between angiographic and computed tomographic imaging of aortocoronary bypasses in 100 patients]. AB - In 100 consecutively operated patients with 167 aortocoronary bypass grafts (ACVB) routine catheterisation was performed three month postoperatively, as well as sequential, ECG-gated and dynamic computed tomography (CT) the day before angiography. The results showed a sensitivity of 92%, a specificity of 67% and an overall accuracy in 81% for CT. Serial (dynamic) CT provides for quantitative assessment of attenuation values of contrast medium enhancement in ACVB and aorta, which improves security of the CT-test in most patients with patent grafts. Therefore clinical contribution of CT will be greatest for routine screening post-operatively and for assessment of ACVB--patency in asymptomatic patients. PMID- 2986218 TI - [Comparison of densitometric and morphometric measurements of global and regional functions using a fully digital subtraction unit]. AB - The value of digital subtraction angiocardiography for evaluating left ventricular movement by a morphometric and densitometric technique was examined by animal experiments (dog). Reversible reduction of global and regional left ventricular function can be quantified by both methods equally well. For judging regional ejection fraction, sector display is more easily comprehended than an ejection-fraction map. For the estimation of global cardiac ejection fraction in 49 patients, morphometric diagnosis was clearly superior to cine cardio angiography (rmorp. = 0.91; rdens = 0.65). For recognising and quantifying regional abnormalities of cardiac movement, semi-quantitative visual gradation is superior to densitometric measurements. PMID- 2986220 TI - [Use of long spin-echo sequences for imaging demyelinating diseases]. AB - A multiple spin-echo-sequence has been evaluated in diagnosis of demyelinating diseases. TE in these MR-experiments was between 14 and 336 msec, TR between 860 and 1660 msec. Three groups of patients were examined, and it could be shown that the probability to detect MS-plaques grows with long TE's up to 300 msec. T2 relaxation-time-values range between 126 and 250 msec. Different MS-plaques in the same patient might show different T2-values. PMID- 2986219 TI - [Nuclear magnetic resonance tomography of the spine]. AB - The present paper deals with the results of 48 MR examinations of known spinal abnormalities. The use of a method strictly related to the individual problem helps to reduce the duration of the examination. Sagittal cuts are particularly useful for demonstrating much of the spinal canal. The spin-echo method is used for all examinations. Demonstration of the cord was performed by using short repetition and echo times (TR, TE). For showing the outer margin of the spinal canal, longer TR and TE should be selected. Surface coils are important for demonstrating spinal disease, since the larger images can be performed without increasing the duration of the examination. PMID- 2986221 TI - [The asterisk sign and adult ischemic femur head necrosis]. AB - The asterisk sign is a stellate density, which is seen normally in the femoral head on computed tomography. It is due to the demonstration of the trabeculae in the femoral head. In adult ischaemic necrosis of the femoral head, there are characteristic changes in the asterisk sign at an early stage, even before there is collapse of the head. The changes are described and the indications for performing CT of the hip for diagnosing adult ischaemic necrosis are discussed. PMID- 2986222 TI - [Scintigraphic research on patellar changes after endoprosthetic replacement of the knee joint]. AB - A clinical and x-ray review of 118 GSB-Knee-Protheses without resurfacing the patella--mean time of follow-up 4 years--was performed. In 50 knees of these an additional quantifying bone-scan with 99mTc-MDP ("ROI-method") indicated new aspects of the pathomechanics of the patella after knee replacement. Comparing 42 healthy knee joints and 82 knees with osteoarthrosis principles for early diagnosis of patellar changes could be set up. According to these findings the quantifying scans show the development of dystrophic patellar changes after knee replacement. PMID- 2986224 TI - [Exo- and endomarginal reactions in the hand in diffuse idiopathic skeletal hyperostosis, their quantification and age dependence]. AB - Diffuse idiopathic skeletal hyperostosis (DISH) may lead to three types of reaction in the hand: increased exomarginal bone formation at numerous fibrous insertions, which increase with age, and endomarginal sclerosis which reaches its maximum during adult life, but which regresses partially with increasing age. The third reaction is increased subchondral sclerosis, which also becomes most prominent in middle age and then diminishes. DISH is more frequently seen in the hand than in the spine and in the hand leads to hyperostosis of varying intensity and in different localities. By quantifying the hyperostosis and comparing it with normal, it is possible to characterise the hyperostosis as it affects the hand. PMID- 2986223 TI - Multifocal, chronic osteomyelitis of unknown etiology. A further report. AB - Four cases of multifocal osteomyelitis of unknown origin in childhood are reported. The variable clinical and radiographic appearances of the disease are illustrated and the diagnostic difficulties in the early stages of the disease are stressed. PMID- 2986225 TI - [Giant-cell reaction of the short tubular bones of the hand and foot]. AB - Giant cell reactions in the short tubular bones comprise mainly enchondromas, aneurysmal bone cysts and giant cell tumours. In both cases a history of trauma can be related. We therefore agree with Aegerter and Lorenzo that an intra osseous haematoma due to minor trauma is likely to be of pathogenic significance. PMID- 2986226 TI - [Calcifications after tylectomy and irradiation for breast cancer. Their symptomatic picture and diagnostic value]. AB - In several institutions lumpectomy combined with radiation treatment is being investigated as an alternative method in the primary management of breast cancer (1-5). In 8 of 53 patients, who had undergone this treatment, new calcifications developed after therapy. In 2 of the 53 patients postoperatively persisting calcifications were encountered. The authors report about their preliminary experience in the diagnosis of calcifications after this therapy. PMID- 2986227 TI - [Percutaneous gastrostomy in the rabbit]. AB - Gastrostomy is performed in patients with obstructive carcinoma of the esophagus. As in this operation the peritoneal cavity has to be opened we tried to find in experiments with 11 rabbits whether the percutaneous gastrostomy is a possible alternative. PMID- 2986228 TI - [Aberrant left pulmonary artery in adults]. PMID- 2986229 TI - [The arteria transversa colli in assessing the neurovascular compression syndrome]. PMID- 2986230 TI - [Computed tomographic diagnosis in aggressive fibromatosis of the neck]. PMID- 2986233 TI - Calcaneus lipoma with bone infarct. An anatomico-radiological study. PMID- 2986234 TI - [Pseudo-gas gangrene of the extremity due to posttraumatic air during skiing]. PMID- 2986231 TI - Oesophageal stenosis caused by benign mucous membrane pemphigoid in a patient with an inactive rheumatoid arthritis. PMID- 2986232 TI - [A free concretion in the abdominal cavity]. PMID- 2986235 TI - [Hereditary myeloperoxidase deficiency syndrome: clinical and hematologic features of 10 cases]. PMID- 2986236 TI - Etiology of small cell lung carcinoma. PMID- 2986237 TI - Small cell lung cancer. PMID- 2986239 TI - Small cell carcinoma of the lung: pathological anatomy. PMID- 2986238 TI - The role of surgery in the treatment of small cell carcinoma of the lung. AB - The role of surgical treatment of non-small-cell carcinoma of the lung is controversial. Surgical therapy of small cell carcinoma of the lung has been the subject of criticism for two decades - in contrast to our opinion about the important role of initial surgical therapy in limited disease of this type of lung carcinoma. In a review of the results of surgical therapy in 109 patients with undifferentiated small cell carcinoma of the lung in the period from 1962 to 1979 and an attempt to define the role of the curative effect, we report on 109 thoracotomies after negative preoperative mediastinoscopy and exclusion of hematogenous spread of the cancer. The resection rate (94/109) was considerably lower than in differentiated lung carcinoma. Pneumonectomy (40%) and extended pneumonectomy (27%) were more frequently performed for small cell carcinoma than for differentiated tumors; thus, the rate of lobectomy and segmental resection (28/94 = 30%, and 3/94 = 3%, respectively) was low. When we included the postoperative lethality in the calculation rather than using the life-table method, we found in a follow-up study of 99% of the patients who had undergone resection, in absolute numbers, 3-year survival in 21/93 patients (23%). In the different stages I, II, and III, we noted 3-year survival rates of 32%, 25%, and 14%. Correlation to the N and T stages was N0 (10/29) 35%; N1 (6/34) 18%; N2 (5/30), 17%; N1 + 2 (11/64) 17%; T1 (11/28) 39%; T2 (8/35) 23%; T3 (2/30) 7%; and T2 + 3 (9/34) 14%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986240 TI - Chemotherapeutic results in small cell lung cancer. PMID- 2986241 TI - Intensive chemotherapy with autologous bone marrow transplantation in small cell carcinoma of the lung. PMID- 2986242 TI - Is there a role for immunotherapy in small cell bronchogenic carcinoma? PMID- 2986243 TI - Cytogenetics of human small cell lung cancer. PMID- 2986244 TI - Growth characteristics and heterogeneity of small cell carcinoma of the lung. PMID- 2986245 TI - In vitro studies in small cell lung cancer cell lines. PMID- 2986247 TI - Development of three human small cell lung cancer models in nude mice. AB - The transplantation of seven human small cell lung cancers (SCLC) into athymic nude mice resulted in the development of three tumor lines that are suitable for study of biology and for tests of new drugs and combinations. They were characterized and the response to known drugs was determined. An identical tumor response was observed in the nude mouse system and in the patient in all four comparisons available. PMID- 2986246 TI - Peptide hormone production associated with small cell lung cancer. PMID- 2986248 TI - Diagnostic procedures in small cell lung carcinoma. PMID- 2986249 TI - [Characterization and differentiation of rat preadipocytes in primary culture]. AB - Using a density gradient medium, we isolated homogeneous cell populations from the inguinal tissue of 3-day old rats. In primary culture we obtained, for the first time, the differentiation of 99% of the cells in the presence of a physiological concentration of insulin (10(-9) M). This model closely mimicked the events occurring during normal mammalian adipose development, i.e. a positive change in beta-adrenergic sensitivity, early induction of lipoprotein lipase, expression of the enzymes involved in the triglyceride systems, and the development of responsiveness to glucagon. PMID- 2986251 TI - Captopril-induced hyperreninemia in cholesterol-fed rabbits. AB - The aims of this study were to investigate the potential effects of captopril (CPT)-induced chronic hyperreninemia on atherogenesis, and to describe and quantitate the morphological changes which occur in the juxtaglomerular (JG) apparatus of drug-treated rabbits. Four groups of normotensive New Zealand rabbits were used. Drug control groups were fed regular rabbit chow (Group I), or regular chow supplemented with cholesterol (Group III). Group II animals were fed regular chow and treated with captopril, and Group IV animals were fed the cholesterol-diet and treated with captopril. Daily captopril administration for a period of six months resulted in significantly (p less than 0.001) increased levels of plasma renin activity and blood urea nitrogen. Mean systemic arterial pressure, plasma aldosterone levels, and hematocrits were significantly reduced in the CPT-treated animals as compared to untreated control groups. No effect on atherogenesis was found. Morphometric analysis showed no difference in the size of the glomeruli between the untreated and the CPT-treated, normal-diet groups, however, significant (p less than 0.001) hypertrophy and hyperplasia of the JG complex were observed in all CPT-treated animals. It is concluded that captopril induced reductions in systemic arterial blood pressure and perfusion pressure, in concert with a blocked renin-angiotensin system which interferes with the normal autoregulation of renal blood flow and glomerular filtration, leads to significant morphologic and functional alterations in the kidney of normotensive animals. PMID- 2986250 TI - Lipoprotein lipase and the uptake of lipids by adipose cells during development. PMID- 2986252 TI - Demonstration of non-opioid sigma binding with (d)3H-SKF 10047 in guinea pig brain. AB - A non-opioid binding site to (d)3H-SKF 10047 (N-allyl-normetazocine), the prototypic ligand for the sigma or PCP-like receptor, was demonstrated. The (d) isomer of 3H-SKF 10047 was used to demonstrate a stereospecific low affinity binding site with a Kd of 173nM. It was naloxone insensitive with an IC50 of greater than 10,000nM, which defined it as non-opioid. Traditional mu compounds like morphine and FK 33824 were also inactive, with IC50's of greater than 10,000nM. Kappa compounds such as ethylketocyclazocine and U-50,488H were active as were all of the benzmorphans tested, with butorphanol the least active. The known antipsychotic haloperidol was the most active compound tested, with an IC50 of 11nM. Other antipsychotics which demonstrated activity were chlorpromazine and pimozide. The atypical antipsychotic clozapine was inactive. PMID- 2986253 TI - Effects of substituted dithiocarbamates on distribution and excretion of inorganic mercury in mice. AB - Diethyldithiocarbamate (DDTC) and six of its N,N-disubstituted congeners were evaluated for their relative efficacies in mobilizing and promoting excretion of mercury (Hg++) using mice which had been given 203Hg++; D-penicillamine (PEN) was used as a positive control compound. While none was as effective as PEN when compared on a molar dose basis, significant activity as assessed by reduction of whole body 203Hg burden, was observed following treatment with three of the analogs. The order of effectiveness was PEN greater than N-methyl-N dithiocarboxyglucamine (MDCG) greater than di(carboxymethyl)dithiocarbamate (DCDC) greater than di(hydroxyethyl)dithiocarbamate (DHDC). PMID- 2986254 TI - Effect of cephalosporins on organic ion transport in renal membrane vesicles from rat and rabbit kidney cortex. AB - The effects of cephaloridine and cephalothin on prototypical organic anion (p aminohippurate, PAH) and cation (N-methylnicotinamide, NMN) transport were observed in brush border and basolateral membrane vesicles prepared from rat and rabbit renal cortex. The cephalosporins interacted with both the cationic and anionic transport systems. Cephalothin inhibited PAH transport in basolateral and brush border membrane in both rats and rabbits. Cephaloridine on the other hand inhibited PAH and NMN transport across rabbit basolateral membranes while it showed a lack of interaction with transport systems in rat basolateral membranes. Conversely, cephaloridine inhibited brush border transport of PAH and NMN in the rat but not in the rabbit. These results provide indirect evidence that cephalothin may be secreted across the renal tubule cell in rats and rabbits while cephaloridine may not accumulate in the rat kidney and becomes trapped in rabbit renal tubule cells. The differences in transport effects observed may explain intra- and interspecies differences in susceptibility to cephalosporin nephrotoxicity. PMID- 2986255 TI - The effect of 3-methylindole on superoxide and hydrogen peroxide production and NADPH oxidation by goat lung microsomes. AB - The production of superoxide and hydrogen peroxide and the oxidation of NADPH by goat lung microsomes in the presence of 3MI or nitrofurantoin were investigated. Although nitrofurantoin strongly enhanced microsomal formation of adrenochrome from epinephrine (a superoxide dependent reaction), 3MI did not increase the rate of adrenochrome formation by microsomes. Neither 3MI nor nitrofurantoin stimulated significant increases in microsomal hydrogen peroxide production. 3MI tripled NADPH oxidation compared to controls, while nitrofurantoin caused a nearly twenty-fold increase in NADPH oxidation. It is concluded that production of superoxide, hydrogen peroxide and depletion of intracellular stores of reduced pyridine nucleotides are not significant components of 3MI pneumotoxicity. PMID- 2986256 TI - A serologic epidemiological study of parainfluenza 1, 2, and 3 viruses in beef cattle and calves in Illinois. AB - Beef cattle and calves in four Illinois populations were studied. Serum was tested by the hemagglutination-inhibition (HI) test for antibodies to parainfluenza viruses (PIV) 1, 2, and 3. In all but group 3 a low level of antibody to PIV-1 and 2 was demonstrated. The highest prevalence and incidence of HI-antibodies was found for PIV-3, followed by PIV-1. There may have been a cross reaction between PIV-1 and -3 in Group 1 calves. The low level of HI antibodies to PIV-1 and 2 may have resulted from infections early in life or colostral antibodies that decreased with age. PMID- 2986257 TI - Perturbations of sarcolemmal and microsomal enzymes by amphiphilic lipids and drugs. AB - Sarcolemmal (SL) and microsomal (MC) membranes were prepared from adult canine cardiocytes. SL Na+, K+-ATPase (2.35 mumole/min per mg) was enriched 117-fold over the homogenate and MC rotenone-insensitive NADH cytochrome c reductase (RINCR) was enriched 41-fold. Preincubation of SL with 50 microM arachidonyl-CoA (20:4 CoA) stimulated Na+, K+-ATPase almost 2-fold; 250 microM 20:4 CoA inhibited the enzyme by 85%. However, RINCR was inhibited 80% by only 0.2 microM 20:4 CoA. Thus, each of these myocardial lipid-dependent enzymes showed a different sensitivity to perturbation by lipid amphiphiles. In further experiments, SL preincubated with 50 microM 20:4 CoA + 2.5 mM propranolol (which had no effect alone) exhibited a synergistic inhibition of the Na+, K+-ATPase: The enzymatic activity declined 8.5-fold when compared to sarcolemma treated with 50 microM 20:4 CoA alone. Thus, the presence of lipid amphiphiles may result in greater inhibition of the Na+, K+-ATPase when propranolol is present in the membrane. PMID- 2986258 TI - Characterization of a partially purified Na+,K+-ATPase from dog heart. AB - Ouabain-sensitive Na+,K+-ATPase of isolated membranes represents a biochemical correlate of the "Na+ pump" that is present in intact tissue and is responsible for dissimilar distributions of Na+ and K+ across cellular plasma membranes. The enzyme has been purified from a variety of sources and its properties have been reported. Only a limited number of studies, however, deal with the cardiac Na+,K+ ATPase. We solubilized this enzyme from dog heart with deoxycholate and effected its further purification by NaI treatment. The method yielded an enzyme preparation of high specific activity (140 mumole/mg protein per hr). The following characteristics were noted: (1) pH optima of 7.4 and greater than 9.0 for ouabain-sensitive and -insensitive ATPases; (2) inhibition by Ca2+ and ouabain, the latter effect being allosteric in nature; (3) inhibition by sulfhydryl reagents (N-ethylmaleimide, p-chloromercuribenzoate) of the ouabain sensitive ATPase but not of the -insensitive enzyme activity. All these properties resembled those seen in isolated plasma membranes (sarcolemma), suggesting that the purification procedure did not alter the properties of mono- and divalent interacting sites as well as a digitalis recognition domain of the Na+ pump. These results thus aid in further understanding the regulation of this vectorial pump that is critical in myocardial function. PMID- 2986259 TI - Influence of explantation procedure on the electrical and morphological properties of cultured neonatal rat ventricle cells. AB - The ultrastructure and electrophysiological properties of ventricle cells from newborn rats were studied before and after explantation. The cultured cells were dissociated either with trypsin or with collagenase, the latter enzyme being used with and without stirring with a magnetic bar. The explanted cells were studied 10 hr and 48 hr or more after explantation. At 10 hr after explantation, the cells exhibited fast-rising action potentials, but their myofibrils were disorganized, except for stirred collagenase-dispersed cells, which were also depolarized and inexcitable. At 2 days and later after explantation, all preparations had well-defined sarcomeres and myofibrils oriented in parallel similar to the ventricle before explantation, but the cells showed slow-response action potentials together with spontaneous activity. These findings suggest that the disorganization of myofibrils does not reflect damage to the surface membrane. Moreover, collagenase seems more damaging to the cells than trypsin under similar conditions (comparable periods of mechanical stirring), especially 10 hr after explantation. PMID- 2986260 TI - Adenosine formation. Evidence for a direct biochemical link with energy metabolism. AB - The increase in tissue and coronary effluent adenosine concentration in hearts undergoing net ATP breakdown results from an accelation of adenosine formation and not from an inhibition of adenosine inactivation. Adenosine formation takes place inside intact isolated cells by a pathway distinct from the cell membrane 5'-nucleotidase, which hydrolyzes only extracellular AMP. Both the magnitude and the variation in the rate of adenosine formation in polymorphonuclear leukocytes undergoing ATP catabolism can be accounted for by the properties of a cytosolic 5'-nucleotidase that is also present in heart. This enzyme, which is allosterically activated by ATP-Mg and inhibited by Pi, provides a direct biochemical link between the energy status of the cell and the rate of adenosine formation. The actions of adenosine to dilate coronary arterioles, antagonize the inotropic effect of catecholamines, and reduce sympathetic-nerve firing would ameliorate the original energy imbalance. Adenosine may therefore function in heart and also in brain, skeletal muscle, kidney, and adipose tissue as a "retaliatory metabolite" that protects the cell against excessive external stimulation. PMID- 2986261 TI - A phosphorus-31 nuclear magnetic resonance study of effects of altered thyroid state on cardiac bioenergetics. AB - We have used 31P nuclear magnetic resonance (NMR) techniques to characterize bioenergetic changes in the Langendorff-perfused rat heart accompanying alterations in thyroid state. Cytosolic phosphocreatine and inorganic phosphate concentrations changed significantly in both the hypo- and hyperthyroid groups compared to controls; the calculated phosphorylation potential [( ATP]/[ADP][Pi]) increased by 60% in hypothyroidism and decreased by 60% in hyperthyroidism relative to the euthyroid value of 47 X 10(3) M-1. Creatine phosphokinase (CPK) and mitochondrial ATP synthase rates were measured in the intact tissue using a saturation-transfer NMR method. There were no significant differences in the measured fluxes through the CPK reaction among the three groups (4.24 +/- 1.00 mM X sec-1 for the euthyroid group). Although O2 consumption increased by 46% in hearts from hyperthyroid animals, no change in the measured mitochondrial ATP synthase flux was observed compared to the euthyroid flux of 1.05 +/- 0.11 mM X sec-1. These results suggest that the apparent in situ P/O ratio of mitochondria in hearts from hyperthyroid animals is reduced relative to that in euthyroid controls. PMID- 2986262 TI - Morphometric analysis of calcium-tolerant myocytes isolated from the adult rat heart. AB - The ultrastructure of calcium-tolerant isolated myocytes has been compared with that of intact heart myocytes using morphometric techniques. Isolated myocytes were prepared by the method of Powell et al. [1] from four adult rat hearts, and multiple left ventricular tissue samples were obtained from a further four rat hearts after fixation by Langendorff perfusion. The subcellular-component volumes of myofibrils, mitochondria, nuclei, transverse tubules, sarcoplasmic reticulum, lipid droplets, and cytoplasmic space have been estimated using a point-counting method on thin sections of intact myocardium and isolated myocytes. No significant differences between the two sample types were found. With the use of freeze-fracture, the numerical density and distribution of sarcolemmal intramembrane particles were analyzed by the method of Jones et al. [2]. The intramembrane particle density of isolated myocyte E-faces was very similar to that of myocytes of intact myocardium. However, isolated myocytes showed a slight reduction in P-face intramembrane particle density (less than 10%), although intramembrane particle distribution remained unaltered on both fracture faces. Our findings show that the ultrastructural features of the isolated myocytes closely resemble those of their counterparts in the intact myocardium. PMID- 2986263 TI - Ultrastructure of the sarcoplasmic reticulum in the ischemic myocardium. AB - Fine structural changes in the sarcoplasmic reticulum (SR) in myocardial ischemia, induced by occlusion of the anterior descending branch of the left coronary artery in the canine heart, were studied by the freeze-fracture technique in situ and in vitro and compared to the alterations in Ca2+-stimulated ATPase activity and sodium dodecyl sulfate gel electrophoresis of the isolated SR. Both SR in situ and the isolated SR exhibited the typical intramembranous particles of 70-90 A in freeze-fracture replicas, and the numbers of particles were more numerous in the concave face (PF) than in the convex face (EF). The numbers of particles in the PF were 2748/micron 2 on the average. In ischemia for 1-2 hr, a significant decrease in the numbers of the particles was found in SR in situ, and corresponding changes were noted in the isolated SR. Decreases in Ca2+ stimulated ATPase activity and in the major protein band of ATPase were recognized simultaneously. The close correlation of the changing patterns between the reduction in Ca2+-ATPase and that in the intramembranous particle density during ischemia supports the suggestion that a large part of the intramembranous particles represents ATPase protein itself. The decrease in the particles of SR membrane indicates the degradation of ATPase in the process of ischemic myocardial necrosis. PMID- 2986264 TI - Enhanced relaxation and reduced positive inotropic effects of amrinone in ventricular muscle from cats with subacute heart failure. Implications for drug therapy. AB - Previously, we reported that amrinone increases isometric twitch force but relaxes K+-induced contracture in muscles from normal cat right ventricle. This study evaluated its effects on diseased cardiac tissue. Right-ventricular papillary muscles were obtained from cats with subacute right-ventricular failure (3-14 days after partial pulmonary-artery ligation) and studied in vitro during stimulation (0.5 Hz) and exposure to high-K+ Tyrode solution. Active isometric twitch force and rate of force development (dP/dt) were significantly lower in muscles from hearts with right-ventricular failure compared to control muscles. In addition, while time to peak force was not different, duration of the twitch was significantly longer. In contrast to its positive inotropic actions in control muscles, amrinone (5.3 X 10(-4) M) had no significant effects on twitch force and dP/dt in muscles from failed ventricles. Time to peak force was not changed by amrinone in either group, but unlike its action in control muscle, duration of the twitch was reduced in failed muscle. Amrinone reduced K+ contracture force similarly in both control and failed muscles. Isoproterenol (10(-6) M) significantly increased twitch force and dP/dt and reduced K+ contracture force in both muscle groups. Since amrinone appears to be a phosphodiesterase inhibitor, our data indicate that cyclic AMP (cAMP)-related relaxation processes, but not cAMP-related contractile processes, can be enhanced by phosphodiesterase inhibitors in experimental heart failure. Furthermore, amrinone's reduced positive inotropic effect in failed myocardium suggests that its improvement of ventricular function in patients reflects, in part, enhancement of relaxation. PMID- 2986266 TI - Tumor localization with 18F-2-fluoro-2-deoxy-D-glucose: comparative autoradiography, glucose 6-phosphatase histochemistry, and histology of renally implanted sarcoma of the rat. AB - Rous sarcoma cells were implanted into the kidney of rats. After 5 days of growth the renal tumor was used for comparing histology with glucose 6-phosphatase (G6Pase) enzyme histochemistry (EHC) and 18F-fluoro-2-deoxyglucose (FDG) auroradiography (ARG). It was found that the regions of the kidney tumor that had retained normal kidney structures were devoid of FDG, whereas there was histochemical staining of normal cortical areas. Regions of tumor growth, on the other hand, retained FDG and lacked G6Pase. Necrotic areas did not accumulate FDG. There was a dramatic decrease in the areas of G6Pase activity as a result of tumor infiltration in the kidney. The results show that FDG, currently being evaluated as a tumor detecting radiopharmaceutical indeed accumulates into areas of vital malignant growth, and they indicate that FDG positron emission tomographic (PET) images reveal the true anatomic location of malignant tissue. PMID- 2986265 TI - Identification of calcium-channel receptors in intact animals. AB - In this study, we demonstrate the in vivo labeling by [3H]nitrendipine ([3H]NTD) of peripheral tissues and the brain in Sprague-Dawley rats. Specific binding is decreased in a dose-dependent manner by nifedipine, with a mean inhibitory dose of 2-10 mg/kg (i.p.). Thin-layer chromatography of the particulate-bound radioactivity reveals that the predominant tritiated drug bound in the left ventricle and the cerebral cortex is [3H]NTD, whereas metabolites constitute the main species in the liver. Peak radioactivity is seen at 15 min following an intravenous injection of [3H]NTD. Highly perfused tissues such as the heart, brain, and lung have significant [3H]NTD binding. In contrast to previously reported in vitro studies, [3H]NTD binding is low in the aorta, skeletal muscle, and ileum. This in vivo animal model is suitable for pharmacokinetic and physiological studies of the calcium channel in intact animals. PMID- 2986267 TI - [Clinical, electromyographic and electroneurographic correlations in lesions of the peripheral nervous system]. PMID- 2986268 TI - [A case from practice (36). Astrocytoma grade IV = glioblastoma multiforme]. PMID- 2986269 TI - Critical evaluation of results of extension of indication for surgery for primary bronchogenic carcinoma. AB - Among 3,120 surgically resected cases (1957-1983) in Shanghai Chest Hospital, 1,476 resections (47.3%) were performed under extended indication. Six categories -(1) aged 70-87 years (102), (2) associated with severe impairment of pulmonary function (25), (3) small-cell anaplastic type (143), (4) stage III lesion (1,145), (5) invading carina (29), (6) with disseminated pleural metastasis and effusion (32)--were critically evaluated. For the first four categories, long term survival rates were very encouraging, whereas only technical advancement and short-term results were discussed for the last two. The authors present strategic points significant in availing higher overall operability and hence the overall survival rate, shedding light on the increase of curative potential for lung cancer. PMID- 2986270 TI - Follow-up care at sleep disorders centers: a commitment beyond diagnosis. PMID- 2986271 TI - [The menopause and vascular risk]. AB - The vascular risk associated with the administration of oestrogens is admitted by the majority, denied by some, and is extremely difficult to assess. As a matter of fact, no vascular accident seems to have been published up to the present day with menopausal women treated with natural oestrogens. The analysis of populations permits one to suppose in the absence of large scale prospective and comparative statistics, that a correct case-finding of risk-factors, and regular surveillance should permit the reduction to a minimum of the probability of vascular accidents. PMID- 2986272 TI - [Strategies for the detection of DNA polymorphism in human populations]. AB - We develop here several models concerning strategies for detection of restriction polymorphisms in humans. These different strategies concern the numbers of individuals to test in a DNA collection, the nature of restriction enzyme to utilize, and the length of the probes. For enzyme utilization different strategies are based on (i) the nature of the restriction site; (ii) the mean length of restriction fragments obtained; (iii) the methylated cytosine characteristics in the CG doublets. Each of these approaches are tested in experimental situations. PMID- 2986273 TI - Betamethasone increases the beta-adrenergic receptor density of human polymorphonuclear leukocytes. AB - Prolonged incubation (18h) of human polymorphonuclear leukocytes (PMNs) with betamethasone (BT) (10(-7) M) increased (3H)-DHA binding to human PMN membranes, apparently causing an increase in the number of beta-adrenergic receptors. Inhibition of this effect by cycloheximide (2 micrograms/ml) suggests that it is dependent on protein synthesis. BT had no effect on basal levels of cAMP in PMNs, but synergistically potentiated the increase in cyclic AMP (cAMP) induced by isoproterenol. Propranolol completely abolished the synergistic effect of BT plus isoproterenol, suggesting that the potentiating effect of BT on cAMP metabolism required the activation of beta-adrenergic receptors. Thus, BT increased the number of beta-adrenergic receptors in human PMN membranes and potentiated the cAMP changes induced by beta-agonists. PMID- 2986274 TI - Prevention of leukocyte migration inhibitory factor activity by glucagon. AB - Leukocyte migration inhibitory factor (LMIF) activity was tested before and after glucagon administration both in vivo and in vitro. In vivo glucagon 1 mg i.v. vs saline administration inhibited LMIF production by T lymphocytes in 85.21% patients (p less than 0.01). In vitro glucagon in physiologic (125 pg/ml) and pharmacologic (50 ng/ml) doses increased the migration area vs PPD 250 microL (migration index 0.5127 vs 0.3210; p less than 0.05). These results show a significant inhibitory effect of glucagon upon LMIF activity. We suggest that glucagon acts by enhancement of the intralymphocytic cAMP/cGMP ratio (cyclic adenosine monophosphate/cyclic guanosine monophosphate). PMID- 2986275 TI - Sucralfate and alginate/antacid in reflux esophagitis. AB - The efficacy of sucralfate and of an alginate/antacid compound was compared in a randomized, double-blind 6-week trial in patients with symptomatic, endoscopically confirmed macroscopic reflux esophagitis. Of the 68 patients who completed the study, 36 received sucralfate and 32 alginate/antacid. Significant symptomatic improvement occurred in both treatment groups: almost 70% of the patients became symptom-free or improved. Esophagitis healed completely in 53% of the patients receiving sucralfate and in 34% of the alginate/antacid patients, as measured with endoscopic criteria (p greater than 0.05). Our results indicate that sucralfate seems to be at least as effective as alginate/antacid in relieving symptoms and in healing macroscopic lesions. As a safe, locally active mucosal protecting agent, sucralfate is a promising new drug for the treatment of reflux esophagitis and deserves further trials over longer periods. PMID- 2986277 TI - Retinoids activate superoxide production by polymorphonuclear leucocytes. AB - Retinol and retinoic acid were effective activators of oxygen consumption by human polymorphonuclear leucocytes (PMN) in micromolar concentrations. In contrast, retinyl acetate was ineffective as an activator. Retinol caused activation only after a lag time, the length of which depended on retinol concentration. Oxygen consumption was due to superoxide production by PMN. Superoxide production was observed as superoxide dismutase-inhibitable cytochrome c reduction. Previously, retinoids have been reported to inhibit PMN activation by phorbol myristate acetate, a tumour promoter. This retinoid-induced inhibition of PMN activation has been suggested to be a mechanism by which retinoids may protect against carcinogenesis in animals. However, the retinoid concentrations at which PMN inhibition was reported were much higher than those found to cause activation in this study. We found that retinoic acid slightly inhibited phorbol myristate acetate-activated superoxide production, but only at concentrations that caused activation. In contrast, activation by formyl-Met-Leu-Phe was effectively inhibited at a retinoic acid concentration that did not cause activation by itself. PMID- 2986276 TI - Terminal differentiation of cord blood lymphocytes induced by thymosin fraction 5 and thymosin alpha 1. AB - Cord blood lymphocytes (CBL) have been shown to be functionally immature compared with normal circulating adult lymphocytes (NAL). Differentiation of T cells is associated with changes in surface antigenic markers and in the pattern of purine degradative enzymes. Previous studies have demonstrated that thymosin fraction 5 (TMS-F5) and thymosin alpha 1 (TMS-alpha 1) can induce in vitro differentiation of murine T-cell precursors and human thymocytes. We have investigated the effects of TMS-F5 and TMS-alpha 1 on the pattern of the purine degradative enzymes adenosine deaminase, purine nucleoside phosphorylase, and ecto-5' nucleotidase (5'NT) of CBL and on the phenotypic markers from the OKT series 3, 4, 8 and 11. Other than a significantly reduced level of 5'NT activity (P less than 0.001) and an elevated percentage of OKT4+ cells (P less than 0.01), CBL demonstrated the same immunological and biochemical patterns as NAL. Incubation of CBL with TMS-F5 (150 micrograms/ml) and TMS-alpha 1 (1 microgram/ml) for 40 h caused a significant rise in 5'NT level and decrease of cells positive for OKT4, resulting in a pattern characteristic of NAL. Thus TMS-F5 might induce the terminal differentiation of CBL, and TMS-alpha 1 seemed to be the active component. PMID- 2986279 TI - Cytomegalovirus in breast milk of Swedish milk donors. AB - 81 milk samples collected from 35 donors 3 days to 7 months after delivery were examined for the occurrence of cytomegalovirus (CMV). 30/35 (89%) of the women were seropositive to CMV. The virus was only found in samples from seropositive women collected greater than 8 days to less than 3 months after partus. During this period milk from 23 seropositive donors were tested; 16/23 (70%) excreted CMV. CMV is evidently a very common finding in the milk of seropositive mothers. PMID- 2986278 TI - Antibodies to human T lymphotropic virus type III demonstrated by a dot immunobinding assay. AB - A dot immunobinding (DIB) technique was applied to the demonstration of antibodies to human T-lymphotropic virus type III (HTLV-III). By this technique IgG antibodies to HTLV-III were demonstrated in six of six Swedish patients with acquired immune deficiency syndrome and in 37 of 39 (95%) Swedish homosexual men with persistent generalized lymphadenopathy but in none of 50 Swedish healthy blood donors. Findings obtained by this method showed complete concordance with those obtained by an enzyme-linked immunosorbent assay. All sera positive by DIB were also positive by Western blotting. The DIB assay is simple and well suited for screening of sera for HTLV-III antibodies. PMID- 2986280 TI - An outbreak of coxsackievirus B infection followed by one case of diabetes mellitus. AB - During the fall of 1979, 22/250 Swedish UN soldiers serving in Egypt were hospitalized with fever and gastroenteritis associated with aseptic meningitis. One of the 22 developed insulin dependent diabetes mellitus (IDDM) 10 weeks following the infection. The majority of the 22 patients showed significant titer rise for coxsackievirus B by plaque reduction neutralization test. The serology results indicate that coxsackievirus B4 most likely caused the outbreak. All 22 were also tested for islet cell cytoplasmic antibodies and islet cell surface antibodies and found negative. The individual developing diabetes mellitus had the HLA-DR phenotype 3,4, which is associated with IDDM. PMID- 2986281 TI - Restriction fingerprinting and serology in a small outbreak of B15 meningococcal disease among Norwegian soldiers. AB - In September 1981 a soldier died from meningococcal septicemia in a military camp in Mid-Norway. Soon afterwards one of his room-mates was transferred to a military camp in Northern-Norway where he shared sleeping quarters (room 7D) with 5 other soldiers of whom 2 fell ill with meningococcal disease 1 month later. Throat cultures were obtained from all 128 soldiers at the military camp in Northern-Norway; 41 (32%) harboured meningococci in their throats. The 3 invasive isolates and the isolates from the 4 healthy carriers at room 7D were all group B and type 15 meningococci. However, by DNA fingerprinting we could identify at least 2, probably 3, different individual strains among these 7 isolates. None of these strains were isolated from soldiers outside room 7D. By use of a B15 whole bacterium ELISA method we showed that the levels of antimeningococcal IgG antibodies in the sera of the two cases at room 7D were low (18 and 28 OD units) compared with the mean IgG levels in the sera of their 4 healthy room mates (1150 OD units) and the mean IgG in the sera from all healthy soldiers (472 OD units). PMID- 2986282 TI - Cancer incidence among workers exposed to radon and thoron daughters at a niobium mine. AB - The aim of this study was to investigate the incidence of cancer among 318 male employees of a niobium mining company which was only operated between 1951 and 1965. Many of the workers, especially underground miners, were exposed to the daughters of radon and thoron and also to thorium. The accumulated doses to the workers from short-lived radon and thoron daughters in the mine atmosphere were assessed to be relatively low; up to 300 working-level months. During the follow up period 1953-1981, 24 new cases of cancer were observed compared to an expected number of 22.8. Twelve cases of lung cancer had occurred versus 3.0 expected. Among the 77 miners, 9 cases of lung cancer were observed against 0.8 expected. Associations between the occurrence of lung cancer and exposure to alpha radiation and smoking were found. For the radon and thoron daughter exposure, about 50 excess cases per million person-years at risk per working-level month were observed. PMID- 2986283 TI - [Pseudohypoparathyroidism, calcifying arteriopathy and ischemic skin necrosis]. AB - A patient with longstanding pseudohypoparathyroidism undergoing substitution with dihydrotachysterin, with normal to low serum calcium and phosphorus levels, developed extensive calcification of the subcutaneous tissue and an obliterative and calcified arteriopathy of the small subcutaneous arteries with ischemic skin signs (livedo reticularis, skin infarction and ulcerative necrosis). After stimulation with exogenous parathyroid hormone there was no increase in urinary cyclic AMP and the G-unit was significantly decreased. It was concluded that the patient is suffering from pseudohypoparathyroidism type 1a. The likely pathophysiological mechanisms and the therapeutic implications are discussed. PMID- 2986284 TI - [Seroepidemiologic study of the occurrence of enzootic bovine leukosis in Switzerland using agar gel immunodiffusion and ELISA in blood and milk serum]. PMID- 2986285 TI - The release of acetylcholine. PMID- 2986286 TI - Coupling of Tetrahymena ribosomal RNA splicing to beta-galactosidase expression in Escherichia coli. AB - Splicing of the Tetrahymena ribosomal RNA precursor is mediated by the folded structure of the RNA molecule and therefore occurs in the absence of any protein in vitro. The Tetrahymena intervening sequence (IVS) has been inserted into the gene for the alpha-donor fragment of beta-galactosidase in a recombinant plasmid. Production of functional beta-galactosidase is dependent on RNA splicing in vivo in Escherichia coli. Thus RNA self-splicing can occur at a rate sufficient to support gene expression in a prokaryote, despite the likely presence of ribosomes on the nascent RNA. The beta-galactosidase messenger RNA splicing system provides a useful method for screening for splicing-defective mutations, several of which have been characterized. PMID- 2986287 TI - Individual tumors of multifocal EB virus-induced malignant lymphomas in tamarins arise from different B-cell clones. AB - Cotton-top tamarins were inoculated with sufficient Epstein-Barr virus to induce multiple tumors in each animal within 14 to 21 days. The tumors consisted of large-cell lymphomas that contained multiple copies of the Epstein-Barr virus genome and generated Epstein-Barr virus-carrying cell lines showing no detectable consistent chromosomal abnormality. Hybridization of tumor DNA with immunoglobulin gene probes revealed that each lymphoma was oligo- or monoclonal in origin and that individual tumors from the same animal arose from different B cell clones. Thus the virus induced multiple transformation events in tamarins in vivo to cause malignant tumors resembling the Epstein-Barr virus-associated lymphomas of patients with organ transplants. PMID- 2986288 TI - Vaccinia virus recombinant expressing herpes simplex virus type 1 glycoprotein D prevents latent herpes in mice. AB - In humans, herpes simplex virus causes a primary infection and then often a latent ganglionic infection that persists for life. Because these latent infections can recur periodically, vaccines are needed that can protect against both primary and latent herpes simplex infections. Infectious vaccinia virus recombinants that contain the herpes simplex virus type 1 (HSV-1) glycoprotein D gene under control of defined early or late vaccinia virus promoters were constructed. Tissue culture cells infected with these recombinant viruses synthesized a glycosylated protein that had the same mass (60,000 daltons) as the glycoprotein D produced by HSV-1. Immunization of mice with one of these recombinant viruses by intradermal, subcutaneous, or intraperitoneal routes resulted in the production of antibodies that neutralized HSV-1 and protected the mice against subsequent lethal challenge with HSV-1 or HSV-2. Immunization with the recombinant virus also protected the majority of the mice against the development of a latent HSV-1 infection of the trigeminal ganglia. This is the first demonstration that a genetically engineered vaccine can prevent the development of latency. PMID- 2986289 TI - Chromatin structure and de novo methylation of sperm DNA: implications for activation of the paternal genome. AB - The chromatin structure characteristic of constitutively expressed genes, tissue specific genes, and inactive genes is absent in chicken sperm chromatin. However, point sites of undermethylation in sperm DNA within constitutively expressed genes, but not within globin genes or an inactive gene, correspond to the location of regions of altered chromatin structure (hypersensitive sites) in somatic tissue and spermatogonial cells. A de novo methylation process whereby regions within and flanking these genes become methylated, but which excludes the methylation of sequences within hypersensitive sites, occurs between the spermatogonial stage and the first meiotic prophase. These undermethylated regions may play a role in the activation of the paternal genome during embryogenesis. PMID- 2986290 TI - Major glycoprotein antigens that induce antibodies in AIDS patients are encoded by HTLV-III. AB - Antibodies from the serum of patients with the acquired immune deficiency syndrome (AIDS) or with the AIDS-related complex and from the serum of seropositive healthy homosexuals, recognize two major glycoproteins in cells infected with human T-cell lymphotropic virus type III (HTLV III). These glycoproteins, gp160 and gp120, are encoded by the 2.5-kilobase open reading frame located in the 3' end of the HTLV-III genome, as determined by amino terminus sequence analysis of the radiolabeled forms of these proteins. It is hypothesized that gp160 and gp120 represent the major species of virus-encoded envelope gene products for HTLV-III. PMID- 2986292 TI - Endogenous anticonvulsant substance in rat cerebrospinal fluid after a generalized seizure. AB - Cerebrospinal fluid taken from rats subjected to electroshock-induced seizures and injected into the cerebral ventricles of rats that had not been shocked increased the seizure threshold of the recipients. The anticonvulsant activity of the donor cerebrospinal fluid was antagonized by opioid antagonists and enhanced by peptidase inhibitors. These results suggest the existence of an endogenous anticonvulsant substance in rat cerebrospinal fluid, possibly opioid in nature, which is activated as a consequence of a seizure and which may play a critical role in postseizure inhibition. PMID- 2986291 TI - Virus envelope protein of HTLV-III represents major target antigen for antibodies in AIDS patients. AB - In this study, two glycoproteins (gp160 and gp120) that are encoded by human T cell lymphoma virus type III (HTLV-III) were the antigens most consistently recognized by antibodies found in patients with the acquired immune deficiency syndrome (AIDS) and with the AIDS-related complex (ARC) and in healthy homosexual males. The techniques used to detect the glycoproteins were radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (RIP/SDS-PAGE). Although most antibody-positive samples from ARC patients and from healthy homosexual males also reacted with the virus core protein p24, less than half of the AIDS patients revealed a positive band with p24 under the same conditions. The ability to detect antibodies against a profile of both the major env gene encoded antigens and the gag gene encoded antigens suggests that the RIP/SDS-PAGE may be a valuable confirmatory assay for establishing the presence or absence of antibodies to HTLV-III in human serum samples. PMID- 2986294 TI - [Herpes simplex and varicella-zona viral disorders. Virologic review]. PMID- 2986293 TI - Radiology and pathology of malignant fibrous histiocytomas of the soft tissues: a report of ten cases. AB - Malignant fibrous histiocytomas (MFH) are believed to originate from histiocytes and are composed of malignant cells with spindle or round shapes. We evaluated ten MFH of the soft tissues by plain roentgenograms, computed tomography (CT), and angiography and subdivided them into four grades of anaplasia and five predominant histologic variants. The variants of MFH demonstrated different vascular patterns. The extension of the lesions could be determined by CT and angiography. CT is the method of choice in the assessment of size and extent of MFH of the soft tissues. When intra-arterial chemotherapy is indicated the angiograms obtained at each catheter placement may substitute CT in follow-up studies obviating additional diagnostic procedures. PMID- 2986295 TI - [Herpes]. PMID- 2986296 TI - Poland-Mobius syndrome: evaluation by computerized tomography. AB - The Poland-Mobius syndrome is the combination of two rare congenital disorders and has been reported previously in only 13 patients. We report two additional cases with an evaluation of the findings and possible pathogenesis, and discuss the role of computerized tomography in evaluating the brain, pons, orbits, and facial nerve. PMID- 2986297 TI - Pain due to nerve injury. AB - Some of the chronic pains that follow disc rupture, myelography, and discectomy may be due to injury to peripheral nerves or nerve roots. The neural mechanisms underlying these pain syndromes are discussed and possible etiologies examined. The roles of peripheral and central changes in neuronal activity and connectivity are explored: plasticity in the nervous system may either be the cause of pain in the 5% of people who develop chronic pain after nerve injury or what prevents pain in the 95% who do not become painful after nerve injury. More research on the behavior of damaged nerves and their central connections is essential. PMID- 2986298 TI - Spread and distribution of human T-cell leukaemia virus type I-reactive antibody among baboons and monkeys in the northern and eastern Transvaal. AB - Antibodies which probably indicate infection with human T-cell leukaemia virus type I (HTLV-I) were determined in three species of non-human primates from several localities. A significant prevalence among chacma baboons and vervet monkeys was confirmed. According to sero-epidemiological evidence that HTLV-I infection is predominantly sexually transmitted between adult animals; comparison of prevalence rates between localities or species should therefore take sexual maturity into account. It appears unlikely that transmission from non-human primates to humans is frequent. PMID- 2986299 TI - Distribution and possible spread of human T-cell leukaemia virus type I in human communities in the northern and eastern Transvaal. AB - The prevalence of serum antibodies which probably indicate infection with a human T-cell leukaemia virus type I was determined among random population samples of more than 100 healthy black individuals in several localities in Transvaal. The percentage of seropositive subjects increases northwards and eastwards, where geoclimatic conditions are similar to those of endemic areas elsewhere in the world. The comparatively higher prevalence among females in the Kruger National Park suggests that this is predominantly a sexually transmitted disease. PMID- 2986300 TI - Comparison of the antihypertensive effect of enalapril and propranolol in black South Africans. AB - Angiotensin-converting enzyme (ACE) inhibitors are useful antihypertensive agents. Enalapril maleate is a new ACE inhibitor with actions similar to those of captopril but with fewer side-effects. A study was conducted on 19 black South Africans with mild or moderate essential hypertension; enalapril was compared with propranolol as monotherapy or together with hydrochlorothiazide in a 1-year randomized, double-blind, parallel study. Neither enalapril nor propranolol alone produced consistent, significant reductions in blood pressure. There were no significant differences between the blood pressure responses to enalapril and to propranolol (either with or without hydrochlorothiazide). It is concluded that neither enalapril nor propranolol is effective as monotherapy in the treatment of hypertension in South African blacks, but that both require the addition of a thiazide diuretic. PMID- 2986301 TI - HTLV-I-reactive antibody in the northern and eastern Transvaal. PMID- 2986302 TI - Erythrocyte pyrimidine-5'-nucleotidase activity as a sensitive indicator of lead exposure. AB - Since the red cell enzyme pyrimidine-5'-nucleotidase (PN) is inhibited by lead, we examined the feasibility of using the activity of this enzyme as a measure of lead exposure. Erythrocyte PN activity was measured in 110 blood samples obtained from subjects working in industries which utilize lead and 40 control subjects. The measurements were then compared with a number of traditional indices of lead poisoning. These included blood and urine lead concentrations and free erythrocyte protoporphyrin, erythrocyte zinc protoporphyrin, urinary coproporphyrin and urinary delta-aminolaevulinic acid levels. There was a highly significant negative correlation between erythrocyte PN activity and blood lead concentration (-0,83; P less than 0,0001), which was greater than that for any of the other measurements. It was therefore concluded that erythrocyte PN activity is an excellent indicator of lead exposure. PMID- 2986303 TI - The action of itraconazole and ketoconazole on growth and sterol synthesis in Aspergillus fumigatus and Aspergillus niger. AB - Growth and sterol synthesis of Aspergillus fumigatus and A. niger were studied in control cultures and in the presence of ketoconazole or itraconazole, the latter compound being 100 times more growth inhibitory than the former. Sterol synthesis is inhibited more rapidly than any visible fungal outgrowth. This inhibition results in an accumulation of 4,14 dimethyl- and 4,4',14 trimethylsterols. The presence of these membrane-disturbing sterols may result in a pertubation of membrane-bound enzyme systems such as chitin synthase. PMID- 2986304 TI - The Tikhoff-Linberg procedure: report of ten patients and presentation of a modified technique for tumors of the proximal humerus. AB - The Tikhoff-Linberg resection is a limb-sparing surgical option to be considered for bony and soft-tissue tumors in and around the proximal humerus and shoulder girdle. Careful selection of patients whose tumor does not involve the neurovascular bundle in the axilla is required. The distal clavicle, upper humerus, and part or all of the scapula are resected. The tumor remains covered by the deltoid muscle plus portions of the muscles that arise from or insert into the resected specimen. In patients with tumors of the proximal humerus a custom prosthesis is used to maintain length and stabilize the distal humerus. Elbow flexion plus stability of the shoulder without the need of an orthosis may be achieved with muscle transfers. Function of the hand and forearm after Tikhoff Linberg resection should be near normal. Review of results in 10 patients shows no local recurrences and excellent function. The major postoperative problem was nerve palsy. The Tikhoff-Linberg procedure should continue to be used for limb salvage in selected patients with tumors in or around the shoulder girdle. PMID- 2986305 TI - Thyrotropin regulation of adenylate cyclase activity in human thyroid neoplasms. AB - Thyrotropin (TSH), stimulators of guanyl nucleotide regulatory protein, (sodium fluoride and guanyl-5'-yl-imido-diphosphate [Gpp(NH)p]) and a stimulator of the catalytic unit of adenylate cyclase (AC) (forskolin) were used to probe the TSH receptor-guanyl nucleotide regulatory protein-cyclase unit in normal and neoplastic thyroid tissue from 17 patients. Eleven of these patients had benign follicular adenomas and six patients had differentiated thyroid carcinomas. An 8000 X g particulate fraction that is rich in thyroid plasma membranes was prepared, and the activity of AC was determined by the conversion of alpha-32P ATP to P32-cAMP. Thyroid neoplasms had a greater AC response to TSH than did normal thyroid tissue removed from the same patients (p less than 0.001). The AC response to NaF and Gpp(NH)p was greater in the neoplastic thyroid tissue, although in these experiments the increase was not significant. In contrast, the AC response to forskolin was comparable in normal (573 +/- 129) and neoplastic (526 +/- 132) thyroid tissue (mean +/- SEM). The effects of NaF, Gpp(NH)p, and forskolin on AC activity were additive with TSH when used at concentrations for optimal AC activity. Low concentrations of NaF and Gpp(NH)p stimulated AC activity whereas high concentrations of NaF and Gpp(NH)p assayed either together or separately inhibited AC activity. When forskolin and NaF were assayed together there was a greater than additive effect or potentiated effect on activity. Basal AC activity was increased in the presence of manganese (Mn+2) (2 mM) over magnesium (Mg+2) (2 mM) (p less than 0.001), whereas TSH-stimulated (p less than 0.01) and Gpp(NH)p-stimulated AC activity (p less than 0.05) were lower in the presence of Mn+2 than Mg+2. There was an excellent correlation between basal AC activity and AC activity in response to forskolin in both normal and neoplastic thyroid tissue, whereas there was no correlation between basal AC activity and TSH-stimulated AC activity in the thyroid neoplasms. These data suggest that the abnormality responsible for the greater AC response to TSH in neoplastic thyroid tissue is proximal to the catalytic unit of AC and most probably is due to an alteration in the guanyl nucleotide regulatory protein or in the coupling of the guanyl nucleotide regulatory protein to either the receptor or the catalytic unit of AC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986306 TI - Unexpectedly high incidence of cytomegalovirus infection in apparent peptic ulcers in renal transplant recipients. AB - Gastroduodenal ulcers in renal transplant recipients are usually accepted as being acid-peptic in origin. In a series of 573 renal transplant recipients there was histologic material available for examination from eight patients with gastroduodenal ulcers and three patients with gastric erosions. All ulcers had originally been diagnosed as peptic or stress ulcers. However, on review, five of them proved to contain cytomegalovirus (CMV); CMV was also present in all the stomachs with erosions. We suggest that CMV infection plays as important a role in upper gastroduodenal ulcers and erosions in renal transplant recipients as it does in similar lesions elsewhere in the gastrointestinal tract of these immunosuppressed patients. PMID- 2986308 TI - [Herpes genitalis]. PMID- 2986307 TI - The interference of naloxone hydrochloride in the teratogenic activity of opiates. AB - Diamorphine hydrochloride, methadone hydrochloride, and the synthetic enkephalin analogue FK 33-824 are potent teratogens for the central nervous system in mouse embryos. They induce the "neurotropic syndrome of malformations," which is restricted to the central nervous system if administered during the critical period of neural tube closure. Pretreatment with corresponding equimolecular doses of the antagonist naloxone hydrochloride applied 30 minutes before treatment with the opiate agonists abolishes the major severe malformations, i.e., exencephaly, craniorachischisis, and brachyury, and reduces the number of cases of kinking of the spinal cord. Dilation of the fourth brain ventricle remains unaffected. It is suggested that the mechanism of interference in the teratogenicity of the opiates by naloxone hydrochloride reported here is based on competition for opiate receptors. In general, these observations are regarded as evidence that the pharmacological affinity of opiate agonists to receptors in the central nervous system is responsible for the malformations caused by them in this system. PMID- 2986309 TI - Deaggregation of human platelets aggregated by thrombin. AB - Human platelets that have undergone the release reaction do not deaggregate readily. We examined conditions under which washed human platelets can be deaggregated after they have undergone an extensive release reaction induced by thrombin (1 or 5 U/ml). To make fibrinogen receptors unavailable, either CP/CPK (or apyrase) was used to remove released ADP, or PGE1 was used to increase cAMP. Chymotrypsin was used to digest proteins that might link platelets, and heparin to interact with released proteins and interfere with their binding to platelets and to each other. Individually, none of these caused deaggregation; heparin did not inhibit the effect of thrombin because no antithrombin III was present. Platelets exposed to thrombin (1 U/ml) which was neutralized at 90 sec by hirudin, could be deaggregated by combinations of CP/CPK (or apyrase) and chymotrypsin, or PGE1 and chymotrypsin. When a higher concentration of thrombin was used (5 U/ml) these combinations caused platelets to deaggregate only when heparin was added before thrombin induced the release reaction. Thus, when extensive release occurs three mechanisms may come into play to link human platelets: one that requires the fibrinogen receptor; a heparin-sensitive reaction that may involve the binding of released proteins; and a linkage that can be disrupted only by proteolysis, providing the other two mechanisms are also inhibited. PMID- 2986310 TI - Maternal and fetal platelet responses and adrenoceptor binding characteristics. AB - Paired blood samples were obtained from mothers (venous) and babies (cord venous blood) at the time of delivery by caesarean section under epidural anaesthetic. Fetal platelets failed to aggregate in response to adrenaline in vitro although adrenaline could potentiate the threshold response to adenosine diphosphate (1 microM). Fetal platelet responses to collagen and 8 Arg vasopressin did not differ significantly from maternal responses. Maternal and fetal platelets also showed similar inhibition of aggregation after activation of adenylate cyclase (PGE1 and parathormone), in contrast to the inhibition of adenylate cyclase by adrenaline. Alpha 2 adrenoceptors were investigated using [3H] yohimbine binding receptor number and were reduced modestly but significantly on fetal compared to maternal platelets. The failure of fetal platelet aggregation in response to adrenaline appears to be related to a failure of receptor coupling and may represent a delayed maturation of fetal platelet alpha receptors or a response to increased circulating catecholamines during birth. PMID- 2986311 TI - Preparation of thrombomodulin from human placenta. AB - Thrombomodulin, a cell surface cofactor for thrombin-catalyzed activation of protein C, was isolated from human placenta by a combination of affinity chromatography and gel filtration. Molecular weight of purified human thrombomodulin, estimated by sodium dodecyl sulfate electrophoresis after reduction, was 88,000. Its isoelectric point was around pH 4. The thrombomodulin was soluble only in the presence of detergent. The purified thrombomodulin was inactivated by disulfide bond reduction, but was stable to heat, pH extremes and protein denaturants. The thrombomodulin served as a cofactor of thrombin catalyzed protein C activation in human as well as bovine enzyme systems. PMID- 2986312 TI - Effect of cetiedil on platelet aggregation and thromboxane synthesis. AB - Cetiedil was found to inhibit platelet aggregation and thromboxane synthesis induced by thrombin and arachidonic acid. When platelets were activated by thrombin, half maximal inhibition (ED50 effective dose of cetiedil necessary for 50% inhibition) for platelet aggregation was 100 microM while that for thromboxane B2 (TXB2) production was 50 microM. When arachidonic acid was used, the ED50 for platelet aggregation was 100 microM while that for TXB2 production was 150 microM. The presence of calcium ions did not affect on the inhibitory effects of cetiedil. The cAMP level in platelets did not increase after incubation with cetiedil. Cetiedil appears to inhibit the activation of platelets related to thromboxane synthesis. PMID- 2986313 TI - [Psychotic episodes following a training seminar in alternative medicine]. PMID- 2986314 TI - [Plasma lipids and plasma lipoproteins]. PMID- 2986315 TI - [Endogenous opioids]. AB - The discovery of the opioid receptors and their corresponding ligands, the opioid peptides, is of fundamental importance as regards our understanding of a variety of functional mechanisms in the central and peripheral nervous system. This review considers the distribution of the multiple opioid peptides within the organism as well as their tissue-specific enzymatic processing. These parameters differ considerably between species. The multiple opioid peptides are paralleled by multiple opioid receptors. These opioid systems affect a broad spectrum of functions, such as behaviour, pain perception, the cardiovascular system, respiration, appetite, gut motility and secretion, water- and electrolyte balance and the complex field of endocrine mechanisms. The task of future pharmacological research is to study these functions by means of more selective opioid agonists and antagonists. An awareness of our present knowledge and of probable findings to come suggests novel therapeutic possibilities in the field of veterinary science. PMID- 2986316 TI - Effects of enalapril maleate (MK-421) on renovascular hypertension. AB - The effects of enalapril maleate (MK-421), a new angiotensin converting enzyme inhibitor, were studied on 5 patients with renovascular hypertension (RVH) due to unilateral renal artery stenosis. The therapeutic dosage was increased when the blood pressure (BP) was not controlled for more than 3 days. Blood sampling was performed before, and 5 hr and 24 hr after the first administration, and on the 3rd day with each dosage. The BP was normalized on 5 mg/day in 1 case, 10 mg in 1 case, 20 mg in 2 cases, and 40 mg plus mefruside in 1 case. Plasma renin activity (PRA) was significantly increased after 5 hr and recovered after 24 hr with 2.5 mg of the enalapril maleate, when the BP was not affected. This indicates that the increase in PRA is likely due to the reduced negative feedback of angiotensin II. When the blood pressure was lowered, PRA was increased and plasma aldosterone concentration (PAC) was decreased significantly. This rise of PRA may depend not only on the reduced negative feedback but also on the fall of BP. It is also considered that the PAC was decreased through the decrease in plasma angiotensin II. A fall of the glomerular filtration rate in one case and also a fall of the perfusion of the kidney of the stenotic side in another case were observed by radioisotope renograms. MK-421 administration was a useful treatment for RVH, and clearly normalized the BP of all the patients studied. However, there was a risk of a fall of renal function on the stenotic side due to the decrease in perfusion pressure. PMID- 2986317 TI - Effect of chronic administration of acetaldehyde by inhalation on (Na+ + K+) activated adenosine triphosphatase activity of rat brain membranes. AB - The effect of chronic acetaldehyde inhalation on (Na+ + K+)-ATPase (EC 3.6.1.3) activities of subcellular fractions of the rat cerebral cortex was studied. Chronic administration of acetaldehyde by inhalation for 4-21 weeks caused significant increases in the enzyme activities of both the synaptosomal plasma membrane (SPM) fraction and the microsomal (MC) fraction. This indicates the change in neural membrane functions of the brain after acetaldehyde treatment. Mg2+-ATPase activities of both subcellular fractions remained unchanged after acetaldehyde treatment. PMID- 2986318 TI - Enterotoxin-induced fluid accumulation during experimental salmonellosis and cholera: involvement of prostaglandin synthesis by intestinal cells. AB - Challenge of rabbit intestinal loops with Salmonella typhimurium or Vibrio cholerae resulted in significant elevation of mucosal cyclic adenosine monophosphate (cAMP) and prostaglandin concentrations. This effect could be reproduced in vitro by exposing either isolated epithelial cells from normal rabbits or Chinese hamster ovary cells to purified cholera toxin or cell-free lysates of Salmonella. Indomethacin treatment of animals challenged with Salmonella resulted in less fluid accumulation, in addition to lower concentrations of intestinal cAMP and prostaglandins, compared to that of similarly changed loops in normal animals. Likewise, intestinal loop challenge of indomethacin-treated rabbits with V. cholerae or purified cholera toxin also resulted in decreased fluid secretion and diminished levels of tissue cAMP and prostaglandins. Intestinal loop tissue from normal animals challenged with V. cholerae displayed 10-fold higher levels of prostaglandins than tissue from uninfected animals. Prostaglandin levels in Salmonella-infected intestinal loops were similarly elevated, but by only 2-3 fold. Chinese hamster ovary cell monolayers were simultaneously exposed to either 10 ng of cholera toxin or 20 microliter of Salmonella lysate and varying concentrations of indomethacin ranging from 1 ng/ml to 10 micrograms/ml. Indomethacin decreased both cAMP and prostaglandin levels in Chinese hamster ovary cells. At high indomethacin concentrations, the cells lost their ability to respond to stimulation with either cholera toxin or Salmonella lysate, while retaining greater than 95% viability as determined by trypan blue exclusion. The prostaglandin and cAMP content of epithelial cells from Salmonella-challenged loops was increased in crypt epithelial cell fractions. Prostaglandin concentrations were elevated in isolated intestinal epithelial cells exposed to purified cholera toxin in vitro. These data indicate that prostaglandins synthesized by the epithelial cells are involved in the pathogenesis of both experimental cholera and salmonellosis. The data are consistent with an enterotoxin-mediated mechanism for both diarrheal diseases and argue against the role of inflammatory cells as the source of elevated cAMP and prostaglandins appearing in the intestinal mucosa during experimental salmonellosis. PMID- 2986319 TI - Lipid accumulation in the livers of chlorpromazine-treated rats does not induce peroxisome proliferation. AB - Treatment of rats with 25 mg/kg/day of the neuroleptic drug chlorpromazine for periods of 7, 28 or 90 days causes a slow accumulation of lipid in large droplets in centrilobular hepatocytes. There is little or no damage to hepatocytes as assessed by changes in glucose-6-phosphatase activity and by electron microscopy. Furthermore there is no indication of a change in peroxisomal beta-oxidation of fatty acids or in microsomal omega-oxidation of fatty acids. It is, therefore, clear that lipid accumulation in the liver does not automatically induce peroxisomal and microsomal fatty acid oxidising enzymes. PMID- 2986320 TI - Lead exposure and changes in the renin-angiotensin-aldosterone system in man. AB - We have studied the effects of chronic slight to moderate lead exposure on the renin-angiotensin-aldosterone system in normotensive man. Plasma renin activity (PRA), blood angiotensin I (AI) concentration, angiotensin converting enzyme (ACE) activity and aldosterone concentration were measured in 33 normotensive men, 25 of whom were occupationally exposed to lead. Positive exponential relationships were found between blood lead and PRA, AI, ACE and aldosterone. Linear relationships were observed between PRA and AI, and between AI and ACE. PRA, AI, and ACE did not significantly correlate with aldosterone. Chronic lead exposure enhanced PRA with a resulting increase in AI levels, which in turn may have produced a substrate-induced increase in ACE. The effects of lead on aldosterone may be mediated in part by an independent pathway. These changes may contribute to the development of hypertension in chronically lead-exposed man. PMID- 2986321 TI - [Lymphographically detectable long-term changes caused by endolymphatic radionuclide therapy (ELRT) in the infradiaphragmatic lymph system]. AB - In sixteen patients who had been treated 42 months before on an average for malignant melanomas of the lower extremity (stage I) by a unilateral endolymphatic therapy with 32P/131J-Lipiodol UF, bipedal lymphography with oily contrast medium was performed. In consequence of the high-dosed intralymphatic radiotherapy the lymph nodes of the treated side are markedly reduced in size and number; fourteen out of the sixteen patients, however, showed lymph nodes of normal size and structure which had not been coloured during ELRT. On the basis of this phenomenon, a possible mechanism of formation of metastases in previously treated lymph nodes is discussed. The presentation and extent of radiogenic reactions in the lymph vessels correspond to the changes known for percutaneous irradiation. PMID- 2986322 TI - Isolation of herpes simplex virus from corneal discs of patients with chronic stromal keratitis. AB - Herpes simplex virus (HSV) was isolated from the corneal discs of six patients with herpes simplex keratitis, which were removed during penetrating keratoplasty and then cultured in vitro. The demonstration of persistent infection in stromal keratitis, even when the condition is quiescent, emphasizes the need for antiviral agents in treatment. The findings support the possibility that latent infection may become established at a peripheral site. PMID- 2986323 TI - Rotavirus infection in children presenting with acute gastroenteritis Ibadan, Nigeria. AB - Faecal samples from 84 children presenting with acute gastroenteritis at the University College Hospital, Ibadan, Nigeria and 28 from a non-diarrhoeal control group were tested for rotavirus by an ELISA. 18 cases (21%) and none of the controls were positive. Hospital in-patients had a higher percentage of positives (25%) than did out-patients (10%). The age distribution of infections showed that the highest infection rate (67%) occurred in children four to six months old. Percentages of positive samples from other age groups was as follows: 0 to 3 months, 13%; 7 to 12 months, 33%; and 13 to 24 months, 16%. None of the faecal extracts from children older than two years was positive for rotavirus. PMID- 2986324 TI - Antibodies to Lassa and Lassa-like viruses in man and mammals in the Central African Republic. AB - 715 human sera from different locations in the Central African Republic were tested by indirect immunofluorescence against the three African arenaviruses, Lassa, Mobala and Mopeia. Four were positive for Lassa, seven for Mobala, and one for both Lassa and Mopeia. The epidemiology of African arenaviruses is discussed. PMID- 2986325 TI - Epstein-Barr virus, immunodeficiency, and B cell lymphoproliferation. PMID- 2986326 TI - Complement activation associated with active cytomegalovirus infection in renal transplant patients, and its absence in transplant rejection episodes. AB - In 20 patients with a cadaveric renal allograft, serial measurements were made of the serum complement factors C3, C4, factor B (FB), and C3d, the stable conversion product of C3. Measurements were started immediately before transplantation and continued thereafter once a week to investigate whether these assays help to differentiate between acute allograft rejection (R) and an active cytomegalovirus (CMV) infection. Fifteen patients had one or more R episodes, and 9 patients suffered from an active CMV infection. Six patients had an R episode and subsequently a CMV infection 13-64 days after R. No significant changes were found in the levels of C3, C4, and FB during R or CMV infection. C3d levels remained unchanged or decreased slightly during R. However, there was a 43-500% increase in the C3d level during CMV infection. This difference in the behavior of levels of C3d during R and CMV infection is significant (P less than 0.01), and suggests that serial measurements of C3d may be useful in differentiating CMV infection from R after renal transplantation. PMID- 2986327 TI - Augmentation of graft-versus-host reaction by cytomegalovirus infection resulting in interstitial pneumonitis. AB - The severity of the graft-versus-host (GVH) reaction, judged by splenomegaly and immunosuppression, was augmented by murine cytomegalovirus (MCMV) infection. Profound GVH-induced immunosuppression was seen in adult unirradiated MCMV infected F1, mice even after challenge with extremely low doses of parental spleen cells. Mice receiving MCMV+GVH challenge died from days 16-21, with interstitial pneumonia being the most prominent pathological lesion. Pulmonary disease was unrelated to levels of viral replication in the lung. These results suggest that in human marrow recipients, cytomegalovirus infection may play a primary role both in provoking or accentuating GVH disease, as well as in the development of interstitial pneumonia. PMID- 2986328 TI - [ATPase activity of the novocaine segregation zones isolated from the erythrocytes of the common frog]. AB - Novocaine segregation zones in frog's erythrocytes, isolated by differential centrifugation, were shown to be ATPase active. The enzyme displays half of its maximum activity at 0.18 Mm ATP concentration to be inhibited by high concentrations of ATP. ATPase is activated by both Mg2+ and Ca2+ (in a lesser degree), with the maximum activity being at pH 7.5. A 5 minutes heating without the substrate results in decreasing the enzyme activity at 30 degrees, and in the total inhibition at 50 degrees C. Along with ATP, the enzyme can hydrolyse GTP and, in a lesser degree, ADP and sodium pyrophosphate. The ATPase activity is not effected with oligomycin (0.5-1.5 mkg/ml) or ouabaine (0.1 mM). Oligomycin in concentration 5 micrograms/ml induced non-specific inhibition of ATPase. Uncouplers, like 2,4-dinitrophenol and carbonyl cyanid p trifluorometoxyphenylhydrazone, stimulate the enzyme activity. The lack in the ATP-ase sensitivity to oligomycin (specific inhibitor of mitochondrial F1-ATPase) and ouabaine (specific inhibitor of Na+, K+-ATPase) may suggest that the ATPase activity of novocaine segregation zones in frog's erythrocytes is not associated with a random contamination with mitochondria or cytoplasmic membranes. The ATPase under study has much in common with the lysosomal +H-ATPase. The results obtained support a hypothesis that +H-ATPase may function as a course of protones for maintaining acidic medium in segregation zones and promote accumulation of weak bases by means of their protonation. PMID- 2986329 TI - [Activity and thermal stability of the enzymes of energy support in the common frog]. AB - The activity and heat resistance of succinate dehydrogenase (SDG), Na, K-ATPase and Mg-ATPase of the grass frog (39 specimens) have been determined. No correlation was found between individual levels of heat resistance both of either enzyme examined and of the same enzyme but taken from different tissues (SDG of liver and muscles). The average level of heat resistance of SDG in liver is significantly higher than that in m. gastrocnemius. A statistically significant correlation was observed between individual levels of enzyme activity in the internal organs (SDG of liver, Na, K-ATPase and Mg-ATPase of kidney). The activity of SDG of muscles does not correlate with that of either partner. PMID- 2986330 TI - [Isoproterenol stimulation of the repair of single-stranded DNA breaks in Chinese hamster cells induced by gamma irradiation]. AB - The action of the increased intracellular content of adenosine monophosphate (cAMP) in CHO-K1 cells (clones 773 and ADr112eb), treated with isoproterenol, on gamma-induced DNA single-strand breaks repair has been investigated. The hormonal treatment stimulates gamma-induced (180 Gr) DNA single-strand repair during the post-irradiation incubation (45 min) by 75 +/- 16%. The results show the involvement of the cAMP system in radiosensitivity of cultured mammalian cells and in regulation of cellular mechanisms of DNA repair. PMID- 2986331 TI - [Electron microscopic analysis of lymphocyte fibrillar elements during the redistribution of surface receptors]. AB - Filaments 5 nm thick, located throughout the cytoplasm mainly along the surface, are observed in intact lymphocytes. In the control glycerinized lymphocytes, besides the above filaments aggregations of filaments nearly 3 nm in diameter were found. After the treatment of cells by antimurine serum or ferritin conjugated concanavalin A, some fibrillar structures are observed mainly in the cap region in the form of filaments 5-6 nm of thickness, radially directed towards the plasma membrane. After glycerinization, three types of filaments are observed being, respectively, near 3, 5-6 and almost 8 nm in diameter. Two latter types of filaments are decorated by S1-myosine fragments which indicates their actine nature. Differences in the character and distribution of myofibrils in the cytoplasm of intact cells and cells with caps may witness in favour of their involvement in the processes associated with redistribution of surface receptors. PMID- 2986333 TI - Signet-ring cell carcinoma of the bladder and the prostate. Report of 4 cases. AB - During a 9-year period in our clinic 4 patients were seen with signet-ring cell carcinoma. 2 patients had primary signet-ring cell carcinoma of the bladder, 1 of the prostate. In the 4th case the tumor was located near the penoscrotal angle of the urethra. These cases are reviewed and their main characteristics, mentioned in the literature, are discussed briefly. PMID- 2986332 TI - [Malignant trophoblastic disease. A clinical study]. PMID- 2986334 TI - [Malignant fibrous histiocytoma of the prostate capsule]. AB - This report deals with the clinical, ultrasonographical, computertomographical and morphological aspects of the rare malignant fibrous histiocytoma of the prostatic gland. A 80-year-old patient suffered from a fast growing painless tumor of the prostatic area, which, on microscopical examination, proved to be a malignant fibrous histiocytoma. The patient refused surgical treatment and died several weeks thereafter. On autopsy no metastases were found. The diagnosis of a malignant fibrous histiocytoma of the prostatic gland can only be made on microscopical grounds. The differential diagnosis includes a pleomorphic rhabdomyosarcoma or a pleomorphic liposarcoma. The clinical differential diagnosis includes, first of all, a carcinoma of the prostatic gland. Surgery is the therapy of choice. The prognosis of the malignant fibrous histiocytoma of the prostatic gland is poor. PMID- 2986335 TI - Specificity of renotropin receptors in murine Wilms tumor model. AB - Contralateral kidneys, removed forty-eight hours after unilateral nephrectomy, contain renotropic growth factors with different effects on normal kidney and Wistar/Furth rat Wilms tumor. Receptors on tumor cells bind factors which, when eluted followed by in vivo bioassay, stimulate the growth of Wilms tumor. Receptors on normal kidney cortex cells bind factors which, when eluted, produce kidney hypotrophy in nontumor-bearing rats. PMID- 2986337 TI - Infectivity of enzootic bovine leukosis infected animals during the incubation period. AB - Steers and calves were experimentally infected with bovine leukosis virus. The virus was isolated from the blood and from the tracheal and bronchoalveolar washings before antibodies could be detected in the serum. Bovine leukosis virus was not detected during any period in the blood plasma. PMID- 2986336 TI - Fibrous histiocytoma of penis. AB - A case of penile fibrous histiocytoma in an eight-year-old boy is reported. This case was treated surgically. Benign fibrous histiocytoma of the penis is rarely reported in the literature. PMID- 2986338 TI - Antibody responses and virus reisolation in turkeys experimentally infected with an infectious bursal disease isolate. AB - Following the infection of turkey poults with a field isolate of infectious bursal disease virus, antibody levels were examined and reisolation of the virus was attempted. After inoculation at 36 days of age, peak titres in both the inoculated and a contact-exposed group were obtained after 13 days. The titres fell slightly during the next week and then remained level until the experiment was terminated at 91 days of age. Virus was reisolated from faeces from day 3 until day 8 after inoculation in the inoculated group and from day 4 until day 9 in the contact-exposed group. In the inoculated group, virus was recovered from the bursae, spleens and intestines on both days 4 and 6 after inoculation, but the thymuses on day 6 only. No clinical signs were observed. PMID- 2986339 TI - Parvovirus and reduced fertility: no link. PMID- 2986340 TI - First isolation of bovine stomatitis papulosa virus in Israel. PMID- 2986341 TI - Isolation of Fasciola hepatica metacercariae by density gradients. AB - Fasciola hepatica metacercariae were purified in high yield, removing contaminating cyst walls and plant material by step gradients consisting of 10 ml of 60% Percoll (density = 1.08 g ml-1) and 10 ml of 50% Metrizamide (density = 1.25 g ml-1). Greater than 90% of the metacercariae applied to the density gradients were recovered. These isolated metacercariae had an in vitro excystment rate of greater than 80%, which was the same excystment rate as metacercariae not subjected to density gradient centrifugation. PMID- 2986342 TI - Metabolic energy pathways in Theileria annulata sporozoites and their significance in sporozoite-bovine lymphocyte interactions in vitro. AB - The existence of metabolic energy pathways has been studied in extracellular T. annulata sporozoites using chemicals known to inhibit specific energy-generating pathways, and their role during invasion of bovine peripheral blood lymphocytes (PBL) by the sporozoites determined in an in vitro system. An inverse relationship was depicted between the dose of various chemicals and the number of T. annulata sporozoites invading PBL: as the concentrations of the inhibitor drugs increased, the number of T. annulata sporozoites within the lymphocytes decreased. An ultracytochemical study demonstrated the presence of the respective pathway marker enzymes, i.e., lactic dehydrogenase (LDH) in the cytosol and within mitochondria, succinic dehydrogenase (SDH) on the mitochondrial membranes and in the contiguous matrix, and cytochrome oxidase (CO) between the inner and outer mitochondrial membranes, in infective T. annulata sporozoites fixed in situ within whole salivary glands of 3-day fed Hyalomma anatolicum anatolicum ticks. PMID- 2986343 TI - [Roentgenomorphological correlations in small-size peripheral lung cancer]. PMID- 2986344 TI - [Glomangioma of the lung (Barre-Masson tumor)]. PMID- 2986345 TI - [Participation of coronaviruses in the etiology of respiratory diseases in calves]. AB - Studied was the Bovine corona virus as taking part in the epizootiology of respiratory diseases in calves. On 4 out of the 7 investigated farms there was a rise of the antibodies against the virus. Two strains were isolated from 2 diseased calves, using tissue culture of trachea. They were the III 213/81 and III 214/81 strains and were identified as bovine corona viruses. Test calves experimentally infected via the trachea with the III 214/81 strain responded with a slight and transient rise of body temperature and a slight cough. The virus was reisolated from the nasal discharge of 3 diseased calves as well as from the feces of one affected calf. The direct immunofluorescence method was employed to establish cytoplasmic fluorescence between the 4th and the 8 h day of infection. The immune response observed with the infected animals confirmed the development of the corona-virus infection. The clinical observations with calves on both the farms with spontaneous infections and those with experimentally induced infections showed that the bovine corona virus could not cause a strongly manifested respiratory disease. PMID- 2986346 TI - [Pathology in hybrid pig breeding due to mixed infections]. AB - A total of 7,500 samples from 5 swine breeding complexes with 200,000 hybrid Camborough pigs were studied morphologically and bacteriologically. The raising technologies used on all complexes were of industrial type or semi-industrial and extensive. A description is given of the simultaneous participation of the following nosologic units: colibacteriosis, responsible for 13 to 14 per cent of the total mortality rate in newborn pigs; bronchopneumonia--causing 6 to 39 per cent losses in the other age groups; and dysentery with salmonellosis--inflicting 5 to 9 per cent losses. Underlined is the great advantage of the industrial technologies of animal raising as against the conventional ones in the prevention of these diseases in hybrid pig breeding. Tested were effective programmes for the prophylaxis associated with the individual nosologic units. More essential were the following: active immunoprophylaxis with live attenuated vaccines against colibacteriosis and salmonellosis; elimination of the alimentary stress factor in dysentery; disinfections of premises by gassing with a mixture of ammonium nitrate and chlorinated lime in a 1:2 ratio. However, all measures proved effective when the principle of "all in, all out' was observed which is essential part of the Camborough industrial raising technologies. PMID- 2986347 TI - [Thymidine kinase activity of virulent and vaccinal strains of Aujeszky's disease virus]. AB - Studied was the timidine-kinase activity of the A2 virulent strain and the MK-25 and MK-35 vaccinal strains of the Aujeszky's disease virus via the infection of primary cell cultures of chick embryo fibroblasts and of pig kidney. It was found that the A2 strain substantially enhanced the induction of the timidine-kinase enzyme in the infected cell cultures, while the IUDR- and the BUDR-resistant vaccinal strains MK-25 and MK-35 showed 10 to 12 times lower activity. Strain MK 25 did not restore the high timidine-kinase activity of the parent A2 strain after 10 successive passages in primary chick embryo fibroblast cultures. The strongly reduced timidine-kinase activity of the two vaccinal strains (MK-25 and MK-35) could be used as a marker to control their properties. The virulent A2 strain of the Aujeszky's disease virus had a well manifested mutability when passing in cultures in the presence of IUDR. PMID- 2986348 TI - [Binary ethyleneimine as an inactivator of the foot-and-mouth disease virus]. AB - Experiments were carried out to inactivate F.M.D. viruses with the use of binary ethylene-imine. It was found that inactivation was optimal when the agent was used at the rate of 0.003 M for 18 hours at 26 degrees C. Its neutralization in the virus suspension was carried out with 3 mM sodium thiosulfate. The inactivated F.M.D. viruses retained their complement-fixing and immunogenic properties. Discussed are the advantages of using binary ethyleneimine as an inactivating agent as against other agents of the asiridine order. PMID- 2986350 TI - Angiomatoid malignant fibrous histiocytoma. Evidence for the histiocytic origin of tumor cells. AB - The results of an histological, immunocytochemical and electron microscopic study of an angiomatoid malignant fibrous histiocytoma are reported. Our results support an histiocytic, rather than an endothelial origin for the tumor cells. PMID- 2986351 TI - [The degree of differentiation and the clinical course of ovarian cancer]. AB - Clinical course versus morphological data on the level of tumor cell differentiation was studied in 323 cases of cancer of the ovaries. Level of cell differentiation proved to be of prognostic value, since it appeared to influence prognosis and clinical course greatly. Metastasis developed in 58.2 +/- 4.5% of patients suffering well-differentiated ovarian cancer. In cases of moderately- and poorly-differentiated cancer, it was 76.1 +/- 5.1 and 90.8 +/- 2.5%, respectively. The high level of tumor cell differentiation was registered in 41.8 +/- 4.5% of patients with cancer at early stages (I and II), while the low--level -in 9.2 +/- 2.5% only. This was matched by the predominance of poorly- differentiated tumors over those with high level of cell differentiation (90.8 +/ 2.5 and 58.2 +/- 4.5%, respectively). The five-year survival rate in patients with poorly--differentiated tumors was 8.3 +/- 2.5%, i. e. 7.8 times lower than in cases of tumors with high level of cell differentiation. PMID- 2986352 TI - [Malignant thecoma with extensive metastasis (a case report)]. PMID- 2986349 TI - Are hepatolithiasis and cholangiocarcinoma aetiologically related? A morphological study of 12 cases of hepatolithiasis associated with cholangiocarcinoma. AB - A few cases of cholangiocarcinoma (CC) related to hepatolithiasis have been reported to date, but the aetiological relationship remains unclear. In an attempt to clarify the relationship between two phenomena, we examined morphologically 12 cases of hepatolithiasis associated with CC and 26 cases of hepatolithiasis without CC, with the aid of immunohistochemical staining for carcinoembryonic antigen (CEA). In the livers where both hepatolithiasis and CC were found, the carcinoma spread along the lumenal surface of the stone containing bile ducts and invaded the ductal walls. Features of "chronic proliferative cholangitis" which was a basic feature of hepatolithiasis, were found within the bile duct walls where carcinoma was invading. In some cases of chronic proliferative cholangitis with hepatolithiasis in the absence of CC, atypical epithelial hyperplasia was noted. Atypical epithelial hyperplasia was also found in bile ducts adjacent to and remote from CC. Atypical epithelial hyperplasia was positive for CEA. The data lead us to speculate that chronic proliferative cholangitis in the presence of hepatolithiasis can undergo progressive changes to atypical epithelial hyperplasia which may in turn progress to CC. PMID- 2986354 TI - [Various hemodynamic and humoral indicators in the hypertensive crisis]. PMID- 2986355 TI - [Use of transcutaneous electric stimulation in diseases of the peripheral nervous system]. PMID- 2986353 TI - Human monoclonal antibodies against group A red blood cells. AB - A human monoclonal antibody with anti-A specificity was generated by Epstein-Barr virus transformation of lymphocytes isolated from splenic tissue after in vitro stimulation with group A red blood cells. This antibody is of the IgM class and directly agglutinates group A red blood cells. Type A1, A2, Aint, A3, AX, Aend and A5 cells were agglutinated by the reagent indicating a single determinant is shared by these A subgroups. PMID- 2986356 TI - [Isolation of high titers of the Schmidt-Ruppin strain of Rous sarcoma virus in a cell culture]. PMID- 2986358 TI - [Effect of the herpes simplex virus antigen on E rosette formation by the lymphocytes of patients with ocular herpes]. AB - Herpes simplex virus antigen was demonstrated to inhibit E-rosette formation in most cases of herpetic eye infection in contrast to those of adenovirus eye infection or healthy donors in whom inhibition of spontaneous E-rosette formation in the presence of herpes virus antigen is observed much more rarely. It is suggested that inhibition of E-rosette formation is associated with specific sensitivity of lymphocytes of the subjects under study to herpes antigen. It is also suggested that spontaneous E-rosette formation may be used to determine the degree of T-lymphocyte response to herpes virus antigen. PMID- 2986357 TI - [Influence of mycoplasma on the production of the oncogenic leukemia virus HTLV by HUT-102 T-lymphoblastoid cells]. AB - A suspension culture of human T-lymphoblasts HUT-102 producing type C oncovirus (HTLV) was examined electron microscopically. Control decontaminated HUT-102 cells spontaneously contaminated with M. orale and exogenously infected with M. pneumoniae were examined in ultrathin sections. The variation statistics method demonstrated that in the mycoplasma-infected cells the number of HTLV virions was significantly higher than in control cultures. It is assumed that the increased production of HTLV is due to mycoplasma-induced changes of physico-chemical properties of the cell membrane which facilitate virion budding. PMID- 2986359 TI - [Immunoprecipitation, with an antiserum to ovalbumin, of protein NP from influenza A virus and of glycoprotein C from the herpes simplex type I virus]. AB - Antisera to ovalbumin and to thyroliberin react in radioimmunoprecipitation and radioimmunoassay with NP protein of influenza A virus and C glycoprotein of herpes simplex virus whose mutual amino acid homology does not practically exceed the limits of homology in tripeptides. Both proteins contain a tripeptide area corresponding to thyroliberin tripeptide. PMID- 2986360 TI - [Acquired immunodeficiency syndrome]. PMID- 2986361 TI - [Properties of herpes simplex virus strains isolated from patients with recurrent cutaneous herpes]. PMID- 2986362 TI - [Electrophoretic characteristics of the virus-specific proteins of Powassan virus]. PMID- 2986363 TI - [Defects in the prostaglandin system. II. Familial platelet-prostacyclin receptor defect (Wien-Hietzing defect)--pathogenetic significance for (early) development of atherosclerosis?]. AB - A case is reported of a 10 year-old girl admitted to hospital because of severe intermittent claudication. Occlusion of the left popliteal artery was diagnosed. Since successful surgical intervention the patient has been symptom-free. No causative factor was detected for this premature occlusion. During intensive laboratory check-up an extreme diminution in sensitivity of the platelets to prostacyclin and a receptor defect were discovered. This defect seems to be genetically determined. The extent to which this as yet unreported platelet defect might have contributed to the development of atherosclerosis at such an early age is discussed. PMID- 2986365 TI - [Structure and metabolism of high-density lipoproteins]. AB - Epidemiological studies have established an inverse correlation between the levels of high density lipoproteins (HDL) in the blood and coronary heart disease (CHD). However, no cause-effect relationship has been established for this important discovery, nor are the mechanisms known by which HDL may provide protection. A recent study has established a strong association between the plasma HDL2 level and the ability to clear dietary fat from the circulation. This article discusses recent studies that demonstrate the important role of chylomicron catabolism as a determinant of HDL2 levels in the circulation. PMID- 2986364 TI - [The life history of brain dopamine]. AB - Neurochemical studies in Parkinson's disease have greatly contributed to the understanding of the neurobiology of the meso-telencephalic dopamine (DA) system; in addition, these studies have significantly influenced our concepts regarding the general principles of brain function. The primary role of DA in striatal function can be seen in its ability to initiate complex patterns of motor activity. The nigro-striatal DA system shows in the face of partial damage an extraordinarily high degree of plasticity, i.e. capacity for functional compensation. The two most important mechanisms of plasticity of the nigro striatal DA system are: compensatory activation of the presynaptic remaining DA neurons (through increase in DA turnover); and increase in the number of postsynaptic DA receptors. The DA loss which occurs during normal ageing is not of sufficient magnitude to cause clinically overt Parkinson's disease. On the other hand, the observations pertaining to the Parkinsonian syndrome produced by NMPTP suggest the participation of environmental factors in the aetiology of idiopathic Parkinson's disease. The remarkable results of nigral cell transplants into the striatum of animals with experimental "parkinsonism", as well as the high therapeutic efficacy of DA substitution in patients with Parkinson's disease point toward a neurohumoral, rather than neurotransmitter, function of brain DA. PMID- 2986366 TI - [The tumor markers TPA and CEA in the after care of breast cancer]. AB - 265 patients with operated and irradiated carcinoma of the breast have been followed up by serum Tissue Polypeptide Antigen (TPA) and Carcinoembryonic Antigen (CEA) analysis during a 6 month period. The results have been compared with those of our routine follow-up-program and the clinical observations. It could be demonstrated, that TPA and CEA serum levels cannot be used as indicators for an early detection of recurrent cancer or the beginning of progressive metastases. The results are contrary to those in the literature. PMID- 2986368 TI - Technetium 99m-pyrophosphate scintigraphy in amyloid cardiomyopathy. PMID- 2986367 TI - [Angiotensin converting enzyme in lung diseases]. AB - The aim of the study was to test the validity of ACE-determination for the differential diagnosis of pulmonary diseases. First details of the method, then the determination of the reference scale are discussed. The ACE-activity in the serum in 60 patients with sarcoidosis and 80 patients with other pulmonary diseases was measured. The sensitivity with which sarcoidosis can be established by means of the ACE-activity is to be examined. No positive correlation between the activity of the enzyme and the x-ray staging could be established, but active diseases requiring treatment could be distinguished from inactive forms to an unexpectedly high degree of reliance in our group of patients. The fact that in some cases of other pulmonary diseases--especially those connected with hepatopathy--higher ACE-activity is found as well, reduces the validity of the method. The determination of ACE comes up to all the criterias of a feasible, clinical-chemical method of analysis: practicability, reliance, high sensitivity and accuracy in a selected group, easy repetition. In some cases a typical x-ray result and corresponding clinical examination may provide the diagnosis of active sarcoidosis without further invasive methods of investigation; observation of the course of disease and adjustment of therapy are possible by means of the control of the ACE. PMID- 2986369 TI - [Small and large intestine devascularization (devitalization) and potentials in the therapeutic use of this operative method]. AB - The experimental findings with devitalization of various intestinal segments by means of devascularization showed that the isolated segments left in peritoneal cavity (together with their contents) were gradually destroyed and changed into a slight fibrous residue. The retaining of the up to 180 cm long segment did not endanger the life of experimental animals. This experience was used in the surgical treatment of inoperable tumours in left half of large intestine and in rectosigmoideum. The case report of four successful operations is given together with two cases of invagination of the devitalized part into intestinal lumen. PMID- 2986370 TI - Pharmacological principles in the development of tolerance. PMID- 2986371 TI - Mechanisms of nitrate-induced vasodilatation and tolerance on a biochemical base. PMID- 2986372 TI - Purification and properties of adenosine kinase from rat liver: separation from deoxyadenosine kinase activity. AB - Ion exchange and affinity chromatography techniques, similar to those previously reported for purification of adenosine kinase from human placenta, were applied to purification of rat liver adenosine kinase. The enzyme, purified 400-fold in 41% yield, was homogeneous on SDS-polyacrylamide gel electrophoresis, with a molecular weight of 52000. It specific activity, 18 mumol/min/mg protein, is the highest hitherto reported for this enzyme from mammalian sources. Chromatography on DEAE-cellulose removed about 98% of the phosphorylating activity towards 2' deoxyadenosine present in the initial pH-treated liver extract. The final preparation exhibited only minimal activity (approximately 1.5%) under optimal conditions (pH 7.5) vs 2'-deoxy-adenosine, the lowest yet reported for such a preparation, with a Km of 670 microM, as compared to 0.3 microM for adenosine. The residual activity towards deoxyadenosine is considered an intrinsic property of the purified adenosine kinase and, in fact, phosphorylation of adenosine was inhibited competitively by deoxyadenosine, with a Ki of 70 microM. Competitive inhibition was also exhibited by cordycepin (3'-deoxyadenosine) with a Ki of 150 microM. A more potent competitive inhibitor was tubercidin, the Ki for which was 1.9 microM. PMID- 2986374 TI - The structure of the major immediate early gene of human cytomegalovirus strain AD169. AB - The nucleotide sequence of the major immediate early (IE) gene of human cytomegalovirus strain AD169 was determined. The structure of the gene was examined by nuclease mapping and by sequence analysis of a cDNA clone made from IE mRNA. The gene encodes a spliced molecule of 1736 nucleotides, made up of four exon sequences of 121, 88, 185 and 1342 nucleotides. Three introns (827, 114 and 170n) were located near the 5' end of the gene. A single open reading frame starting in the second exon extends for 491 amino acids, corresponding to a protein of molecular weight 64000. The putative promoter region contains several short direct and inverted repeat sequences of 16, 18, 19 and 21 nucleotides, which extend 509n upstream from the transcription start site. The structure of the major IE gene and its protein product are discussed and compared with the corresponding IE gene from the Towne strain of HCMV. PMID- 2986373 TI - [Cancer through the eyes of the epidemiologist (on the epidemiology of noninfectious diseases)]. PMID- 2986375 TI - Rise per base pair in helices of double-stranded rotavirus RNA determined by electron microscopy. AB - Regular helices of double-stranded RNA occur in nature only as the genome of certain viruses. The structure of such double-stranded RNA helices has been little studied compared to that of DNA, but some X-ray crystallographic data (Arnott, 1970; Saenger, 1984) are available. The recent advent of sequence data of bovine rotavirus RNA (Dyall-Smith et al., 1983; Elleman et al., 1983; Ward et al., 1984) has enabled us to determine by direct measurement of electron micrographs the translation, or axial distance between base pairs in RNA duplexes. Using two different spreading conditions we obtained values of 2.79 +/- 0.10 and 2.80 +/- 0.11 A. These results are consistent with the 11-fold A RNA (Arnott, 1970; Rosenberg et al., 1976) proposed for the conformation of double stranded RNA. We included both circular and linear molecules of phi X174 RF DNA in the same preparations, and the translations for these molecules were between 3.23 +/- 0.06 and 3.29 +/- 0.05 A. Thus, double-stranded RNA contained 1.16 to 1.17 times more nucleotides per unit length than DNA. PMID- 2986376 TI - [Solid sintered calcium hydroxylapatite as a carrier and support for bone growth]. PMID- 2986377 TI - Binding of transforming DNA to protoplasts isolated from competent Bacillus subtilis. AB - Transforming 3H-labelled DNA binds specifically to protoplasts isolated from competent cultures of Bacillus subtilis. The bound DNA is fully accessible to added DNase I, indicating that protoplasts bind, but do not process donor DNA. A bi-phasic pattern of competition for binding with increasing amounts of unlabelled and labelled DNA was found. In these conditions, two levels of saturation appeared which correspond to two kinds of DNA receptor sites. Homologous DNA binds preferentially to the first kind of receptor (specific) and only at higher concentrations would bind to the second (unspecific) binding sites. Bound DNA forms a rather stable complex with some constituent(s) of the protoplasts. PMID- 2986378 TI - [Prevention and therapy of herpesvirus infections]. AB - The group of the human-pathogenic herpesviruses comprises five subgroups: Herpes simplex virus type 1 (HSV-1), Herpes simplex virus type 2 (HSV-2), varicella zoster virus (VZV), cytomegalovirus (CMV), and Epstein-Barr virus (EBV). Primary infection with these ubiquitous herpesviruses usually occurs in childhood or during adolescence and frequently remains inapparent. However, it can also give rise to a variety of clinical pictures. Important clinical manifestations of herpesvirus infections are mucocutaneous lesions (HSV-1, HSV-2, VZV) self limited, lymphoproliferative diseases (CMV, EBV) and congenital malformations (CMV). Primary infection with herpesviruses leads to a persistent infection of the host. This clinically silent condition of latency can be interrupted and may cause pathological symptoms to recur by reactivation of latent herpesviruses. A classical example of the clinical manifestation of herpesvirus reactivation is herpes zoster following an overcome varicella disease. The mechanism of herpesvirus reactivation has not yet been fully clarified. Reactivation of herpesviruses might be attributable to a weakening of the cellular immunodefence. For the control of viral infections mainly two cellular effector systems are responsible: unspecific, cytotoxic, natural killer (NK) cells and specific cytotoxic thymus-dependent (T) lymphocytes. The functional impairment of these cytotoxic active cells my cause herpesvirus reactivation in immunodeficient or immunosuppressed persons. Interference with the immunological control function may also contribute to the genesis of herpesvirus-associated tumours. Such an association between herpesviruses and human tumours is assumed to exist especially in the case of EBV. The frequently life-endangering severity of local or disseminated herpesvirus infections calls for suitable measures ensuring efficient prophylaxis and therapy. However, the possibilities of a specific immunoprophylaxis (vaccine, special immunoglobulins) against herpesvirus infections are still rather limited. The development of antiviral substances has greatly benefited from the introduction of new agents (Acyclovir) and the production of sufficient quantities of interferon (IFN) preparations during the last few years. Impressive results were obtained with the nucleoside-related substance Acyclovir in the prevention and therapy of primary or reactivated HSV-1 or HSV-2 infections. The use of Acyclovir as prophylactic agent produced the effect that recipients of bone-marrow transplants were no longer afflicted by HSV 1 infections.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986380 TI - [Viral hepatitis and immunoprophylaxis]. AB - Today we distinguish 4 forms of viral hepatitis: hepatitis A, hepatitis B, hepatitis non-A, non-B and hepatitis occurring in the course of other viral diseases. The viruses of hepatitis A and hepatitis B have been identified but the agent(s) of hepatitis non-A, non-B remain unknown. Inoculation of normal pooled human immunoglobulin provides passive immunity for 2-3 months against hepatitis A but not against hepatitis B or hepatitis non-A, non-B. For passive protection against hepatitis B a special immunoglobulin with a high anti-HBs titer must be used whereas the protection against hepatitis non-A, non-B with immunoglobulin is uncertain. Live attenuated and noninfectious polypeptide vaccines for active immunisation against hepatitis A are currently developed and first clinical trials have begun with the live attenuated vaccine. A vaccine consisting of noninfectious highly purified HBsAg derived from the plasma of HBV carriers is in general use since two years and has proved safe and highly effective and vaccines are now developed from HBsAg obtained through molecular cloning of the HBsAg genome in plasmids and expression of the genome with HBsAg production in yeast cells. First clinical studies with this vaccine are encouraging and these as well as purely synthetic vaccines will in time replace the currently used vaccines. No vaccines could be developed so far against hepatitis non-A, non-B because the agent(s) of this disease are unknown. PMID- 2986379 TI - [Viral diarrheas]. AB - In the developing countries diarrhoea ranks among the most frequent diseases: 5 18 million children are estimated to die annually from gastro-intestinal infections. But also in Europe and the USA diarrhoea is of utmost medical importance, especially among children and infants. It was only twelve years ago that 2 viral groups, the Norwalk and the rotavirus group, were discovered to be etiological agents responsible for a large proportion of gastro-intestinal infections. Whilst viruses of the Norwalk group cause primarily gastro-intestinal infections in schools and families (school-children and adults), rotaviruses attack mainly infants. In our latitudes rotaviruses are important agents responsible for nosocomial infections. A rapid diagnosis is crucial if the chain of infections is to be interrupted. Uncontrolled spread may not only lead to severe disease in infants and young children, but also burdens health insurance - plans considerably. Experiments intended to develop a rotavirus vaccine are under way. PMID- 2986381 TI - [Vaccination of animals and human health]. AB - Prophylactic immunization of animals against obligat and nonobligat pathogenic zoonoses benefit human health in many ways both directly and indirectly. Typical examples of a direct protective effect are the vaccinations of dogs, cats and foxes against rabies as well as the vaccinations against respiratory diseases in cows, horses, dogs and cats to which the most varied species of pathogens of noncompulsory zoonoses contribute. A considerable contribution to the protection of human health is made by the vaccination against salmonellosis and leptospirosis, against vesicular stomatitis, American equine encephalitis and against other zoonoses spread by arthropods, against ecthyma and stomatitis papulosa as well as against brucellosis, anthrax, Q-fever, Newcastle disease and foot-and-mouth disease. The indirect effects of prophylactic vaccination of animals on human health are very complex and still need investigation. An example of this are the vaccinations of animals against human and animal influenza A viruses which can inhibit hybridisation and recombination between human and animal influenza viruses in an ecological system. Occasionally prophylactic vaccinations of animals can do harm to human health. This is invariably a rare incidence in immuno-suppressed persons caused by live vaccines i.e. prophylactic vaccination against Newcastle disease in fowl or against orthopox in animals by the use of the common vaccinia strains, after compulsory vaccination for humans had been cancelled. Prophylactic vaccinations of animals must be constantly followed up and their action on human health must be checked. In the case of positive results prophylactic vaccinations must be carried out selectively and in a wide range. PMID- 2986382 TI - [Increased sensitivity of the bronchial system of hypertensive pregnant patients to beta-adrenergic stimulation]. AB - Airway resistance at term pregnancy was evaluated by oscillation technique in 15 hypertensive and 15 normotensive late pregnant women, and in 10 non-pregnant women. The recordings were obtained before and 1, 3, 6 and 12 min after administration of 0.6 mg Fenoterol by dose aerosol. - The Fenoterol-induced decrease of airway resistance was twice as large in pregnant women with hypertension as in the other two groups. PMID- 2986383 TI - [Morphological studies of the Leydig cells in impaired fertility]. AB - The histological results of 242 testicular biopsies in the period from 1974 to 1983 were evaluated with special consideration of the Leydig cells. Normal spermatogenesis was found in 9.5% of cases, hypospermatogenesis with and without maturation arrest in 76.4%, also Sertoli-cell-only and Klinefelter's syndrome in 11.6% and 2.5% respectively. Frequently an increase as well as a decrease of Leydig cells were observed in cases with pathological findings in these biopsies. In patients with Klinefelter's syndrome only an increase of Leydig cells could be seen. Ultrastructural investigations have shown that there were degenerative alterations of nucleus and cytoplasm in the Leydig cells of pathological biopsies, which could not be seen with light microscopy. In one patient with Klinefelter's syndrome needle-like crystalloid inclusions of unknown genesis were found in cytoplasm. PMID- 2986384 TI - [DNA repair processes in thymus and spleen cells of rats in vitro as affected by the poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide]. PMID- 2986385 TI - [Cell-mediated cytotoxicity and lymphocyte stimulation in Aujeszky's disease. I. In experimentally infected swine]. PMID- 2986386 TI - [Comparative studies of the use of the enzyme-linked immunosorbent assay (ELISA) for the determination of antibodies against the avian encephalomyelitis agent]. PMID- 2986388 TI - [Cell-mediated cytotoxicity and lymphocyte stimulation in Aujeszky's disease. II. Following vaccination of swine and subsequent infection]. PMID- 2986387 TI - Vaccination confers protection against Aujeszky's disease in cattle. PMID- 2986390 TI - [Taxonomic identification of gram-negative nonfermenting microorganisms in environmental research]. AB - The scheme for the identification of Gram-negative nonfermenting microorganisms is proposed. The scheme comprises the most important key signs, such as the cytochrome oxidase reaction determined by the method of Gaby and Hadley, the oxidation/fermentation test, maltose oxidation and motility, as well as additional key signs, among them gelatinase activity, the oxidation of 10% lactose, nitratase activity with the liberation of free nitrogen, the utilization of the sources of carbon and energy (glucose and sodium acetate) in limited media containing ammonium salts and nitrates as the sources of nitrogen. Additional tests for the identification of nonfermenting microorganisms similar in their main key signs are also recommended. PMID- 2986389 TI - [Comparative research on erythrocyte anionic adenosine triphosphatase in vertebrates]. AB - Studies have been made on the level of activity of anion ATPase and its sensitivity to anions in erythrocytic membranes from fishes, amphibians, reptiles, birds and mammals. Significant variations in these properties of the ATPase were found among the species investigated. Protein composition of erythrocytic membranes was also investigated by means of disc-electrophoresis in polyacrylamide gel in men, rabbit, rat, mouse, hamster, tortoise, crow and starling. PMID- 2986391 TI - [Production of immobilized SalI and PvuII restrictase preparations]. AB - Conditions for the immobilization of specific endonucleases Sal I and Pvu II on BrCN-activated Sepharose 4B have been selected. Some physico-chemical properties of the preparations of immobilized restrictases Sal I and Pvu II have been characterized. The specific and general activity values of the preparations thus obtained have been established. The immobilized enzymes have been used for the multiple restriction of the DNA of phage lambda and the DNA of Neisseria meningitidis. PMID- 2986392 TI - [Iron and the pathogenicity of bacteria]. PMID- 2986393 TI - [Isolation of unbalanced growth forms from Listeria and their ultrastructure]. AB - As the result of the action of penicillin, LiCl and UV irradiation on Listeria culture in the logarithmic phase, unbalanced growth forms (UGF) were obtained. Their viability was determined by the number of colony-forming units per ml. In this investigation the action of the above factors in a single cultivation and in consecutive subcultures was studied. Viable UGF were obtained under the action of all the above factors. UGF obtained under the action of penicillin were found to convert easily into L-forms in the process of subculturing. On the contrary, UGF obtained under the action of UV irradiation and LiCl produced no L-forms and reverted easily when the action of these factors stopped. In all three cases the formation of UGF was accompanied by changes in the structure of the cell wall and, partially, the cytoplasmic membrane, but in each case these changes had different character. PMID- 2986395 TI - [Disorders of nocturnal sleep in Huntington chorea]. AB - A polygraphic study of nocturnal sleep was conducted in 11 patients with Huntington's chorea. It was found that Stage I sleep and the phase of rapid sleep (PRS) were prolonged while delta-sleep was considerably shortened. This was accompanied by an increase in the latent periods of all stages of slow sleep and the shortening of the latent period of PRS. The patients displayed a tendency toward an increased number of sleep spindles and a considerable decrease in the delta-index. Patients with Huntington's chorea showed an increase in body movements during sleep. Dissociations were detected in polygraphic characteristics of individual phases of sleep. The reduction of delta-sleep is related to cerebral cortex atrophy while the increase in PRS is explained by neurotransmitter disturbances characteristic of this disease. PMID- 2986394 TI - [Hepatitis A foci in children's collectives studied by using clinico-biochemical, virological and serological diagnostic methods]. AB - In 4 foci of hepatitis A (HA) at children's institutions in Moscow 218 children and 30 staff members were examined. Simultaneously with clinico-biochemical studies, specific IgM and IgG in the blood and hepatitis A virus (HAV) antigen in feces were determined by radioimmunoassay. Different forms of HA were detected in 29.1% of children in kindergartens, 19.4% of school children and 3.3% of adults, which was due to great differences in the size of the immune stratum among them (IgG to HAV was detected in 8-10% of children aged 3-6 years and in 83% of adults). In 2/3 of children with HA in the foci the disease was not accompanied by jaundice, 1/5 of them having the inapparent form of this infection. In 45% of the patients HAV antigen was detected in feces, irrespective of the form of HA, 5 14 days before the appearance of hyperfermentemia, and in 2/3 of them this antigen was also detected during the first 6 days after that. Simultaneously with an increase in aminotransferase activity, all of the HA patients showed the presence of specific IgM in their blood. PMID- 2986396 TI - [Analgesic effect of high-frequency and acupuncture-like transcutaneous electric stimulation of nerve fibers in spinal osteochondrosis]. AB - Epicutaneous electrostimulation of nerve fibers was used in 192 patients with the neurologic manifestations of spinal osteochondritis in order to alleviate the pain syndrome. Two stimulation regimens were used: a high-frequency one (rectangular impulses of current with an 0.1-0.2 msec duration and a 70 Hz frequency and the intensity being subliminal for a motor response) and "acupuncture-like" a series consisting of seven impulses of the same duration and frequency with the repetition of three series in one second, the intensity of the irritant being over the motor response threshold). The occurrence of prolonged analgesia was shown to be approximately equal with both regimens and to depend significantly on the duration of the pain syndrome and the presence of the symptoms of the loss of spinal root function. There were several cases when after one of the regimens had failed, the pain was controlled by the alternative method of stimulation. PMID- 2986397 TI - [Dynamics of the bioelectrical activity of the brain in patients with intrasellar pituitary adenomas irradiated with a proton beam]. AB - Sixty patients with pituitary endosellar adenomas irradiated with the proton beam were examined using EEG prior to and following irradiation in the period ranging from 14 days to 3 years. Fourteen days to three months after the proton irradiation, intensified irritation of the diencephalic structures was recorded. Later on, gradual normalization of bioelectrical activity was observed. Local changes in the temporal lobes, probably associated with the tissue response to irradiation by the proton beam were noted only in 8 out of the 60 patients. PMID- 2986398 TI - On the self-affinity of heparan sulfates from quiescent or proliferating normal 3T3 cells and from SV40-transformed cells. AB - 35S-Labelled heparan sulfates derived from the culture medium (extracellular), a trypsinate of the cells (pericellular) and the cell residue (intracellular) of quiescent normal, proliferating normal or SV40-transformed 3T3 cells were analyzed for charge heterogeneity, by ion exchange chromatography and for self affinity, by chromatography on heparan sulfate-agarose gels. Quiescent normal cells retained most of their heparan sulphate intra- or pericellularly. The surface-exposed material was charge heterogeneous and had a strong affinity for heparan sulfate. In cultures of growing cells and transformed cells most of the heparan sulfate was found in the medium. The heparan sulfate retained on the surface or growing cells had a lower self-affinity than did the corresponding material from normal and transformed cells. Although cell surface heparan sulfates from transformed cells showed affinity for a matrix substituted with the total heparan sulfate pool, the affinity for one particular subtype was much less pronounced or non-existent. PMID- 2986399 TI - Possible site of negative and positive feedback action of oestrogen on gonadotrophin secretion in normal women. AB - For determination of the site of action of oestrogen (E) during the negative and positive feedback phases of gonadotrophin secretions, studies were made on the pituitary response to a small amount of LRH and the pulsatility of gonadotrophins after E administration in normal cycling women in the mid-follicular phase. The pituitary responses to an iv bolus of 2.5 micrograms of synthetic LRH were evaluated by measuring serum LH and FSH 2 h before and 8 h after administration of 20 mg of conjugated E (Premarin). In the next cycle, the pituitary responses to a same dose of LHRH were also observed 2 h before and 56 h after E injection. The mean levels of serum LH and FSH and the peak responses to LRH were significantly (P less than 0.05) decreased 8 h after E injection, but were significantly (P less than 0.05) increased 56 h after E administration. In the third cycle, the pulsatility of gonadotrophins was evaluated by measuring serum LH and FSH every 15 min for 180 min before and 8 h and 56 h after E injection. The pulse frequencies of gonadotrophins were not significantly different before and 8 h and 56 h after E injection. The amplitudes of pulses 56 h after Premarin injection were significantly higher than those before the injection. These findings suggest that the negative and positive feedback effects of E on gonadotrophin secretion may be caused, in part, by its direct action on the pituitary response to LRH. PMID- 2986401 TI - Urinary adenosine 3',5'-monophosphate (cAMP) response to antidiuretic hormone in diabetes insipidus (DI): comparison between congenital nephrogenic DI type 1 and 2, and vasopressin sensitive DI. AB - Urinary adenosine 3',5'-monophosphate (cAMP) excretion before and after administration of aqueous vasopressin (pitressin) and 1-deamino-8-D-arginine vasopressin (DDAVP) was measured in congenital nephrogenic and in vasopressin sensitive diabetes insipidus (VS-DI). Excretion of cAMP into the urine increased markedly in response to pitressin (676%) and to DDAVP (252%) in VS-DI. Nephrogenic diabetes insipidus (N-DI) could be divided into two categories (type 1 and type 2) in respect to urinary cAMP responsiveness. In type 1, cAMP excretion showed no definite change after stimulation with pitressin (102%) or DDAVP (127%). On the other hand, urinary excretion of cAMP was significantly elevated in response to DDAVP in familial cases of N-DI type 2 (1269%) without producing any concentrating effect on the urine. Two different defects are considered to be involved in the pathogenesis of N-DI. PMID- 2986400 TI - Correlation between the number of thyroliberin binding sites, the tumour size and the plasma prolactin level in human prolactin-secreting adenomas. AB - Basal plasma prolactin (Prl) level, tumour size and [3H]thyroliberin (TRH) binding to tumour membranes were studied in 18 patients bearing Prl-secreting adenomas. Big tumours (grade III) were associated with high plasma Prl levels (median value: 1929 (range: 207-3570) ng/ml) and possessed numerous membrane TRH receptors (median value: 508 (range: 0-1200) fmol/mg of protein). By contrast, smaller tumours (grade II and I) were associated with lower plasma Prl levels (median values: 1085 (range: 40-1890) and 295 (range: 98-788) ng/ml, respectively) and possessed fewer membrane TRH receptors (median values 122 (range: 11-328) and 13 (range: 0-52) fmol/mg of protein, respectively). A direct positive correlation was demonstrated between the plasma Prl level and the number of [3H]TRH binding sites (rho: 0.729 P less than 0.001). That the higher number of TRH receptors is associated with the largest tumours may be of importance in hyperprolactinaemia and should be taken in account when speculating on the pathogenesis of human Prl-secreting adenomas. PMID- 2986402 TI - Studies on the mechanism of acute inhibition of thyroglobulin hydrolysis by iodine. AB - Iodine in excess is known to acutely inhibit thyroidal secretion. In the present study we have characterized the time course of the iodine effect in vitro and investigated the underlying mechanisms. Labelled thyroid glands were cultured in vitro in medium containing mononitrotyrosine, an inhibitor of iodotyrosine deiodinase. The rate of hydrolysis of labelled thyroglobulin was measured as the proportion of labelled iodotyrosines and iodothyronines recovered at the end of culture and was used as an index of thyroidal secretion. Thyrotrophin (TSH) administered in vivo acutely stimulated the rate of thyroglobulin hydrolysis. Addition of NaI to the culture medium acutely inhibited both basal and TSH stimulated thyroglobulin hydrolysis. The effect of iodide was demonstrable after 2 h, maximal after 6 h and was not reversible upon removal of iodide. Iodide abolished the dibutyryl cAMP induced stimulation of thyroglobulin hydrolysis. Iodide required organic binding of iodine for its effect but new protein or RNA synthesis was not necessary. The inhibitory effects of iodide and lysosomotrophic agents such as NH4Cl and chloroquin on thyroglobulin hydrolysis were additive suggesting different sites of action. Iodide added in vitro altered the distribution of label in prelabelled thyroglobulin in a way that suggested increased coupling in the thyroglobulin molecule. These data indicate that 1) the iodide effect occurs progressively over a 6 h period, 2) continued presence of iodide is not necessary once the inhibition is established, 3) iodide exerts its action primarily at a post cAMP, prelysosomal site and 4) the effect requires organic binding of iodine, but not new RNA or protein synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986403 TI - Nuclear reverse T3 binding sites: an artefact of isolation? AB - Binding of [125I]rT3 to rat liver nuclear extracts prepared in 0.25 M sucrose could be abolished by a prior wash of the nuclei with 2.4 M sucrose. Analysis of mixtures containing [125I]rT3 and nuclear extracts (0.25 M sucrose) showed that after incubation for 2 h at 22 degrees C, degradation of rT3 into 3,3'-T2 and I- had taken place. It appears that the presence of 125I- in these mixtures can account for the previously observed 'binding' of [125I]rT3 to these nuclear extracts. Further characterization of the deiodination process in nuclear extracts showed: 1) inactivation by heating, 2) production of equimolar amounts of I- and 3,3'-T2, 3) stimulation by sulfhydryl compounds and inhibition by propylthiouracil in a fashion similar to the microsomal iodothyronine 5' deiodinase (ping-pong mechanism). We conclude that the observed deiodination of rT3 in hepatic nuclear extracts is of enzymatic nature, due to contamination of the nuclear preparation by microsomal iodothyronine 5'-deiodinase. However, since the nuclei are prepared in the presence of the non-ionic detergent Triton X-100, a nucleus associated deiodinase activity cannot be totally excluded. PMID- 2986404 TI - Radioimmunoassay for 21-deoxycortisol: clinical applications. AB - A radioimmunoassay for 21-deoxycortisol is described. The immunogen, 21 deoxycortisol-3-(0-carboxymethyl) oxime-bovine serum albumin, was prepared, the antisera raised against it were studied and the reliability of the assay was checked. The antiserum selected cross-reacted with 11-deoxycortisol (0.08%), corticosterone (0.25%), cortisol (0.6%) and 17-hydroxyprogesterone (1.6%). 21 deoxycortisol was separated by celite partition chromatography and eluted in the 70/30 (v/v) isooctane/ethyl acetate fraction together with 11-deoxycortisol and corticosterone. The radioimmunoassay was used to measure 21-deoxycortisol in the plasma of normal subjects and patients with androgen excess. In normal subjects, men (0.19 ng/ml +/- 0.08) and women (0.18 ng/ml +/- 0.09) had similar basal levels (mean +/- SD). One hour after ACTH stimulation, these levels were increased by a factor of 3.5. In 7 patients treated for classical congenital adrenal hyperplasia associated with 21-hydroxylase deficiency, basal values varied between 9.1 and 39.9 ng/ml (measured at 8 a.m.). In 7 untreated women with late-onset congenital adrenal hyperplasia (with 21-hydroxylase deficiency), ACTH stimulated levels were increased to between 9 and 25.5 ng/ml. In 14 heterozygous carriers of 21-hydroxylase deficiency, diagnosed by HLA genotyping, all ACTH stimulated levels were well above the highest corresponding levels in normal subjects, whereas 17-hydroxyprogesterone levels remained within the normal range in 9 of the cases. PMID- 2986405 TI - Infantile neuroaxonal dystrophy in an albino girl. A cliniconeuropathologic study. AB - We present a cliniconeuropathologic study of infantile neuroaxonal dystrophy (INAD) in a 5-year-old Black girl with albinism. The clinical picture shows progressive psychomotor deterioration, beginning after 1 year of age, with hypotonia, pyramidal signs, optic atrophy, and deafness. Light-microscopic examination of the brain reveals wide distribution of spheroids, cerebellar atrophy, and neuronal loss with astrocytosis. This is the first described case of the combination of INAD with albinism. PMID- 2986406 TI - Polyglucosan bodies in the sural nerve of a diabetic patient with polyneuropathy. AB - In the sural nerve of a 62-year-old woman with impaired glucose tolerance test and polyneuropathy, many intra-axonal polyglucosan bodies were observed. Polyglucosan bodies have been described in spontaneously or alloxan-diabetic rats, but are not usually observed in human diabetic neuropathy. Since intra axonal polyglucosan bodies can occur in the sural nerve in various diseases and in aging, they are considered as non-specific changes. Their presence is probably related to a primary axonal neuropathy. PMID- 2986407 TI - [Selectivity of [D-Ala2-O-benzyl-Ser5]-enkephalin on mu and delta opiate receptor subtypes]. PMID- 2986408 TI - [Effect of total alkaloids of Fissistigma oldhamii on cAMP and isolated hearts]. PMID- 2986409 TI - Detection and characterization of four binding sites for opioids in the mouse brain. AB - The binding sites labelled by 3H-dihydromorphine, 3H-ethylketocyclazocine and 3H D-Ala2-L-Leu5-enkephalin in mouse brain membranes were characterized in cross competition studies. The data was evaluated by simultaneous non-linear least squares regression analysis with the "Ligand" program (Munson & Rodbard 1980), that had to be upgraded to handle more than three binding sites. By statistical analysis four different binding sites were identified. Three of the sites probably correspond to the pharmacologically well characterized mu, kappa and delta-opioid receptors, respectively, and their binding capacities relate as 1:1.5:2.5. Classification of the fourth site is more problematic. Using 3H ethylketocyclazocine it had higher capacity than the others and bound ethylketocyclazocine with a relatively high affinity (Kd = 10 nM), dihydromorphine with a very low affinity (Kd greater than 10(-5)M) and showed no binding of D-Ala2-L-Leu5-enkephalin. In displacement studies, N allylnorcyclazocine (SKF 10,047), though unselective, bound with highest affinity to the mu and the fourth site. Since naloxone did not bind to this fourth site, it can not be termed an opioid site in a strict sense, but it might have some relevance in view of non-naloxone-reversible effects reported for some opioids. PMID- 2986410 TI - Cyclic GMP affects redox state and improves energy charge of ischaemic Langendorff-perfused rat heart. AB - The effects of 8-bromo cyclic GMP (1 X 10(-5)M) on the levels of lactate, NAD+, NADH, AMP, ADP, ATP, creatine phosphate (CrP) and creatine were studied in the ischaemic Langendorff-perfused rat heart. The NAD+/NADH ratio and the energy charge were also calculated. The dependence of the effect of 8-bromo-cGMP on substrate availability was also studied by adding pyruvate (5 mM and 10 mM) to the perfusate, and by comparing the changes to those during perfusion with glucose alone. When glucose (11 mM) was used as the only substrate, 8-bromo-cGMP administration resulted during ischaemia, in a decrease of the NADH level, and in an increase of the energy charge. When pyruvate (5 mM) was added to the perfusate, 8-bromo-cGMP administration resulted in an increase in the NAD+/NADH ratio and in an increase of the energy charge. When the pyruvate concentration was further increased (10 mM), no changes were seen in the beforementioned variables after 8-bromo-cGMP administration. It is concluded that the effects of 8-bromo-cGMP may be exerted by an enhancement of carbohydrate oxidation. PMID- 2986411 TI - Myocardial calcium uptake in streptozotocin diabetic and control rats after dibutyryl-cAMP; alpha-adrenergic stimulation and calcium deprivation. AB - Previous studies have shown a decreased myocardial calcium uptake after beta adrenergic stimulation with isoproterenol in isolated perfused hearts from streptozotocin diabetic rats. Abnormalities in the beta-receptor-adenylate cyclase system could explain this but in order to circumvene the receptor we studied the effect of the permeable cAMP analogue, dibutyryl-cAMP on this calcium uptake. A marked increase was seen in control hearts while no increase could be registered in diabetic hearts. Defects in the protein kinase phosphorylation system or in the protein kinase substrate in the sarcolemma are suggested. alpha Adrenergic stimulation with phenylephrine, being a cAMP independent positive inotropic agent, was also tested but no increase in calcium uptake was seen in either control or diabetic hearts. This could be due to a different effect on calcium currents during action potential after alpha-stimulation compared to the beta-adrenergic effect. Reexposure to calcium after calcium deprivation leads to excessive myocardial calcium uptake (calcium paradox), but the increase was significantly smaller in diabetic hearts, suggesting a differential responsiveness to the damage induced by this procedure. Early biochemical abnormalities in the basement membrane or in the composition and calcium binding ability of the sarcolemma could possibly constitute a common final site for the defect myocardial calcium uptake after isoproterenol, db-cAMP and calcium deprivation in streptozotocin diabetic rats. PMID- 2986412 TI - Ethanol dependent interaction between prostaglandins and lipoxygenase products in human peripheral lymphocytes. AB - The effect of lipoxygenase products, 12-L-HETE and 15-L-HPETE, on cyclic AMP levels in human peripheral lymphocytes was examined in the absence and in the presence of a prostaglandin of the E-type (0.6-3.0 microM) or isoprenaline (33 microM). The studies were performed either in the absence or in the presence of 6 per cent ethanol. For comparison the effect of arachidonic acid and linolenic acid were studied. In the absence of ethanol 12-L-HETE and 15-L-HPETE had no significant effect on cyclic AMP accumulation. However, in the presence of ethanol 12-L-HETE (above 1 microM) inhibited prostaglandin E1 but not isoprenaline induced cyclic AMP accumulation. 15-L-HPETE had a biphasic effect on prostaglandin E2 induced cyclic AMP accumulation. Concentrations below 1 microM stimulated, those above inhibited. Virtually complete inhibition was seen at 15 microM. The two other fatty acids inhibited both prostaglandin E2 and isoprenaline induced cyclic AMP accumulation in the presence, but not in the absence of ethanol. The results show that lipoxygense products have little or no effect on cyclic AMP accumulation in human peripheral lymphocytes unless ethanol is present. In the presence of ethanol both 12-L-HETE and 15-L-HPETE appeared to selectively affect the cyclic AMP accumulation stimulated by PGE. PMID- 2986413 TI - p,p'-DDT-induced myoclonus in mice: the effect of enhanced 5-HT neurotransmission. AB - The acute behavioural consequences of intragastric p,p'-DDT in high doses to mice are stimulus sensitive abrupt muscle jerks (myoclonus). The serotonin (5-HT) precursor 5-hydroxytryptophan (5-HTP) ameliorated in contrast to the natural precursor tryptophan, the neurotoxin-induced myoclonus. The extracerebral decarboxylase inhibitor carbidopa and the selective 5-HT reuptake inhibitor paroxetine both enhanced the antimyoclonic action of 5-HTP. The effect was reversed by the 5-HT receptor blockers cinanserine and methysergide. The data add further evidence to a central serotonergic mechanism involved in p,p'-DDT induced myoclonus. PMID- 2986414 TI - The quantal nature of transmission and spontaneous potentials at the Torpedo electromotor junction. AB - Miniature and stimulus evoked electroplaque potentials (mEpPs and EpPs) were recorded in Torpedo electrocytes intracellularly and extracellularly. The quantal release parameters of EpPs and the time course of quantal EpCs were estimated in normal and low Ca2+-high Mg2+ solutions. Amplitude-frequency distribution of mEpPs showed Gaussian or uneven character with an average mean value of 0.3 +/- 0.08 mV (S.D.). The mean coefficient of variation of mEpPs was 26.8 +/- 7.2% (n = 6). Tetrodotoxin reversibly blocked the stimulus evoked EpP but hardly influenced the amplitude-frequency histogram of spontaneous EpPs in 10(-8)-10(-6) M concentration. The quantum content of stimulus evoked EpPs varied between 100-400 in normal solution which decreased in low Ca2+-high Mg2+ solution and the quantal release conformed to binomial statistics and allowed determination of the parameters p and n. Frequency of the spontaneous discharges varied highly from electrocyte to electrocyte but an analysis of the time intervals showed randomness for the events. The decay phase of quantal current composed of non exponential and exponential sections which was characteristic with 0.75 +/- 0.16 msec (mean, S.D., at 20 degrees C) time constant of exponential decay. Although, two types of mEpCs could be differentiated having significantly slower and faster time courses. Neostigmine prolonged the time constant of decay of mEpCs in dose dependent manner with a factor of 2 in 10(-6) M and of 4 in 10(-5) M concentrations (at about 20 degrees C). PMID- 2986415 TI - The effects of different vitamin D-states on intestinal absorption of vitamin D3 and its metabolites in rats. AB - Rats were kept on a diet deficient in vitamin D for 2 months and were then randomly assigned to one of three groups with different supplies of vitamin D for 9 days. At the end of this period [3H]-cholecalciferol (5 microCi) was administered intragastrically and the serum radioactivity was recorded after various periods of time. The animals were kept in metabolic cages and urine and faeces were collected. After 3 days the animals were killed and the liver, kidney and fat tissue were investigated for radioactivity. The radioactivity was separated into different fractions of vitamin D by means of chromatography. The animals with vitamin D deficiency displayed higher levels of serum radioactivity, which were to a great extent confined to the fractions of the more polar metabolites (25(OH)D3 and 1,25(OH)2D3). There was significantly less radioactivity in the 3-day faecal collection from these animals. In general, there was a low urinary excretion of radioactivity. In the rats with sufficient vitamin D, most of the radioactivity remained as the parent substance and was also detected in increased amounts in the liver, kidney and fat tissue. The results suggest a discriminatory enterohepatic cycling of vitamin D and its more polar metabolites leading to an increased faecal loss of the pro-hormone in animals with a vitamin D surplus. It is proposed that the selectively effective enterohepatic reabsorption of the different metabolites may serve as a protective mechanism when body stores of vitamin D are overloaded. PMID- 2986416 TI - On long-lasting potentiation in the hippocampus: a proposed mechanism for its dependence on coincident pre- and postsynaptic activity. PMID- 2986417 TI - Effect of Althesine on spinal cord activities. PMID- 2986418 TI - On the mechanism of action of theophylline and caffeine. AB - Important developments in our understanding of the mechanism of action of methylxanthines have taken place in the last 10 years. A brief overview of these developments is provided below and the author concludes that the common view that theophylline (and caffeine) acts by raising the levels of cyclic AMP is generally untenable. Instead, many of the actions of the methylxanthines can be explained on the basis of their being antagonists of endogenous adenosine. However, the mechanism behind the antiasthmatic effects of xanthines still remains unknown and further research is necessary. PMID- 2986419 TI - Cushing's syndrome due to an ACTH-producing neuroendocrine tumour in the nasal roof. AB - A patient with ectopic adrenocorticotrophic hormone (ACTH) production from a neuroendocrine tumour of the nasal roof is presented. By indirect immunoperoxidase techniques the tumour cells were shown to be distinctly positive for ACTH and beta-endorphin but negative for other peptides derived from pro opiomelanocortin. Neither corticotropin releasing hormone (CRF) found in some tumours associated with ectopic Cushing's syndrome, nor gastrin immunoreactivity, which coexists with ACTH in normal rat pituitary and in rat and human gastrointestinal cells, were demonstrable in the tumour. A review of other, previously recognized locations of CRF/ACTH producing tumours is given to increase the awareness of the ectopic Cushing's syndrome, which may lack the classical features and is characterized by fulminant clinical course, extreme fatigue, weakness, pale facial swelling, oedema and hypokalaemic alkalosis. PMID- 2986420 TI - Thyroid hormones affect serum angiotensin I converting enzyme levels. AB - Serum angiotensin I converting enzyme (ACE) was measured in 10 patients with Graves' disease and 2 with thyroiditis during different stages of the diseases. The effect of thyroxine on serum ACE levels was also recorded in 12 patients with thyroid cancer, who were on thyroxine suppression. Serum ACE levels correlated positively with clinically assessed thyroid function and peripheral thyroid hormone levels, especially during hyper- or hypofunction. ACE was measured both with an enzyme kinetic and a new, quantitative inhibitor binding assay. The methods gave similar results, which indicates that ACE increments during thyroid hyperfunction were quantitative, and not a result of increased enzyme activity. Serum ACE increments associated with high lysozyme concentrations are signs of immunologic activation or proliferation of monocytic cells. In this study there was no correlation between the two enzymes, which may indicate either increased synthesis or possibly shedding of ACE from endothelial cells or delayed metabolic clearance of this enzyme. Serum ACE measurements may provide a useful tool for assessing thyroid function and the effect of thyroxine treatment. PMID- 2986421 TI - Hypothalamic control of lipid metabolism. AB - Obesity as a common disorder of lipid metabolism might be caused by defective hypothalamic control as demonstrated by ventromedial lesions or the effect of cholecystokinin application. The hypothalamic proopiomelanocorticotropin is the precursor of hormonal fragments affecting fat mobilisation, the endocrine pancreas and gastrointestinal functions. PMID- 2986422 TI - Corticotropin releasing factor (CRF): diagnostic implications. AB - Nine normal volunteers received an intravenous bolus injection of 50, 100, and 200 micrograms ovine Corticotropin releasing factor. There was no dose response relationship between the injected oCRF dosage and stimulated ACTH, beta endorphin, and cortisol secretion. When human synthetic CRF was injected (50 and 100 micrograms i.v.) no significant difference compared to the oCRF induced ACTH stimulation was observed. In contrast to the lacking relationship between the CRF dosage and the biological response there was a clearcut dose response relationship between the amount of oCRF injected and the CRF immunoreactivity measured 15 minutes after injection with a specific oCRF radioimmunoassay. No serious side effects were observed when the 100 micrograms CRF dosage was used as standard dose in the CRF test in patients with diseases of the hypothalamo pituitary-adrenal axis. In patients with Cushing's syndrome the CRF test is helpful for the differential diagnosis (ACTH dependent Cushing's disease, autonomous cortisol secretion due to an adrenal adenoma or carcinoma, ectopic ACTH syndrome). In addition the CRF test is of prognostic value after surgical or neurosurgical therapy of Cushing's syndrome. Furthermore secondary adrenal failure after operative therapy can be documented by the CRF test. In patients on corticoid therapy the degree of suppression of CRF induced ACTH secretion correlated to the dosage and the duration of corticoid therapy. The main suppressive effect of corticoids on the hypothalamo-pituitary-adrenal axis seems to take place at the pituitary level. In patients with secondary adrenal failure the analysis of ACTH secretion after CRF administration allows the differential diagnosis between hypothalamic and pituitary ACTH hyposecretion. In conclusion the administration of oCRF has been shown to be a well tolerated and useful tool in the differential diagnosis of the causes of hyper- and hypofunction of the hypothalamo-pituitary-adrenal axis. Though there was only 10% cross reactivity with synthetic human CRF, CRF immunoreactivity could be detected in 53 out of 55 pregnant females. The results of measuring endogenous CRF levels in patients with diseases of the hypothalamo-pituitary-adrenal axis are preliminary but endogenous CRF levels measured by the heterologous oCRF radioimmunoassay, correlated well to the clinical situation and the ACTH-levels. These results have to be verified with a homologous hCRF radioimmunoassay. PMID- 2986425 TI - Effect of dopamine receptor stimulation on the inhibition of LH pulsatility by a met-enkephaline (FK 33-824). AB - Plasma LH fluctuations reflect pulsatile hypothalamic GnRH activity. In order to investigate a possible interaction between opiate and dopaminergic pathways in the control of LH and PRL release, a met-enkephalin analogue (FK 33-824) was administered intravenously to female volunteers at a rate of 0.01 mg/kg/h for 4 h. LH pulses as recorded by plasma measurements every 20' were significantly (p less than 0.01) inhibited whereas PRL concentrations were increased. Pre treatment with bromocriptine, 1.25 mg b.i.d. for 3 days, counteracted the stimulatory effect of FK 33-824 on PRL secretion but the inhibitory effect on LH episodic release was not modified by this dopamine agonist. Results indicate that opiate receptor stimulation selectively depresses hypothalamic GnRH activity and enhances PRL release from pituitary lactotrops. The GnRH lowering effect does not seem to be mediated by dopaminergic mechanisms which govern PRL secretion. Also transient, opioid-induced hyperprolactinemia is not causally related to the suppression of hypothalamic GnRH activity. PMID- 2986423 TI - Human pancreatic tumor GH-releasing factor. AB - Within the past year, three similar peptides with specific growth hormone (GH) releasing effects have been extracted from human tissue, identified, and synthesized. Human pancreatic tumor GH releasing factor (I-40)-OH (hpGRF-40) was the sole hpGRF isolated from the pancreatic tumor of a patient in Charlottesville and was the predominant peptide isolated from the pancreatic tumor of a patient in Lyon. The Lyon tumor also contained hpGRF(1-37)-OH and hpGRF(1-44)-NH2. Both immunological and biochemical data suggest that hpGRF-40 and hpGRF-44 are present in the human hypothalamus and may be the human GH releasing hormone(s) (GHRH). In cultures of rat pituitary cells, hpGRF stimulates GH but affects neither basal and dopamine-inhibited prolactin release nor basal and gonadotropin releasing hormone (GnRH)-stimulated luteinizing hormone (LH) release. hpGRF stimulates cyclic AMP production within seconds, an effect which is blocked by somatostatin. In contrast, while hpGRF stimulates phosphatidylinositol turnover in the pituitary, the effect is not inhibited by somatostatin. In the human, hpGRF-40 (1 microgram/kg) given intravenously (i.v.) stimulates GH release within 5 minutes. hpGRF-40 does not elevate serum prolactin levels, thyrotropin (TSH), LH, or corticotropin (measured indirectly through plasma cortisol), or blood glucose or plasma concentrations of insulin, glucagon, pancreatic polypeptide, cholecystokinin, gastrin, gastric inhibitory peptide, motilin, or somatostatin. When graded doses of hpGRF (0.1-10 micrograms/kg) are given i.v., no differences are noted in the maximal levels of serum GH achieved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986424 TI - Human pancreatic growth hormone releasing factor (hpGRF): GRF- and GH-levels after bolus injection and infusion of hpGRF. AB - Synthetic human pancreatic growth hormone releasing factor (hpGRF) was given as an i.v. bolus to healthy volunteers in 5 different dosages (3.3 micrograms to 200 micrograms hpGRF). In addition 11 healthy subjects were infused over 2 respectively 5 hours in a dosage of 100 micrograms hpGRF/h after receiving a bolus of 50 micrograms hpGRF. Four healthy subjects served as placebo controls. GH, PRL, TSH, and GRF were measured by specific radioimmunoassays. The results show the clearcut dose response relationship between the administered GRF dosage and the resulting GH response from 3.3 to 50 micrograms hpGRF i.v. Higher dosages of hpGRF did not lead to a more pronounced GH response though there was a linear dose response relationship between the administered hpGRF and the GRF immunoreactivity 5 minutes after injection. Infusion of hpGRF could not sustain elevated GRF levels and a second bolus of 50 micrograms hpGRF given at the end of the 2-respectively 5-hour infusion led to a minor increase compared to the first bolus. 100 micrograms hpGRF was given to 14 patients with active acromegaly leading to a significant rise of the GH levels with the exception of 3 patients. Of the latter 3 two had received previous therapy, and one patient suffered from ectopic GRF hypersecretion. When GH responses to hpGRF were compared to the responses to other releasing hormones there was no correlation. After transsphenoidal surgery divergent responses of GH were seen. In one patient with low basal GH and an exaggerated rise after GRF before surgery there was no response after successful transsphenoidal operation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986426 TI - Endocrinological differentiation of primary hypothalamic and pituitary disease. AB - The paper reviews approaches that can be applied to separating the endocrinological central nervous system (CNS) from pituitary components of neuroendocrine disorders. Major reasons which hamper the differentiation of primary hypothalamic from pituitary diseases are considered. Finally, future strategies suitable for a better accomplishment of this task are mentioned. Theoretically, the combined application of CNS-active compounds and hypothalamic regulatory hormones (RHs) should unravel the hypothalamic or pituitary etiology of the disorder. Until now, the most useful diagnostic application of RHs deals with their use in disease states due to isolated or multiple RH deficiency. Thus, the recent isolation of GRF molecules from human pancreatic tumors holds promise of a better functional characterization of GH deficiency states. The differentiation of the hypothalamic or pituitary origin of the disease in states of pituitary hyperfunction, e.g., acromegaly, or prolactinomas, is influenced by factors such as the development or expression of aberrant pituitary receptors for neurotransmitters and neuropeptides, the persistence of the responsiveness of tumorous pituitary cells to the respective RH, the disrupting effect of the excessively produced pituitary or target gland hormone on hypothalamic function. Future strategies of research in this area may include: 1. the development of sensitive and specific assays for RHs, e.g., GRF, CRF, etc. and RH determinations in biological fluids; 2. the search for non-hypophysiotropic peptides in the cerebrospinal fluid and their evaluation after appropriate stimulation; 3. better functional and clinical characterization of ectopic hormone-producing syndromes, or of psychiatric disorders mimicking neuroendocrine abnormalities from an endocrinological viewpoint (e.g., primary affective disorders v. Cushing's disease; 4. use of neuroactive compounds probing selective aspects of neurotransmitter function and, finally, 5. long-term follow-up studies after adenomectomy. PMID- 2986427 TI - Monoclonal antibodies reactive with breast tumor-associated antigens. PMID- 2986428 TI - Fusion proteins in retroviral transformation. PMID- 2986429 TI - Application of migration inhibition techniques in tumor immunology. PMID- 2986431 TI - Mutation in human populations. PMID- 2986430 TI - Genetic mutations affecting human lipoprotein metabolism. PMID- 2986433 TI - [Retinal centrifugal projection in the dog]. PMID- 2986432 TI - Surgical remodeling of the silhouette by aspiration lipolysis or selective lipectomy. AB - The author, a pioneer in originating the technique of suction lipectomy, discusses in detail the pathophysiology of the fat cell and fat metabolism. In addition, he describes the surgical means by which, since 1977, he has popularized this innovative new approach to the treatment of unsightly fat deposits in many sites in the body, including the thighs, buttocks, abdomen, hips, arms, ankles, breasts, and submental regions. PMID- 2986434 TI - [Ca2+-dependent cysteine proteinase (calpain) in bovine lens. Degradation of lens structural proteins]. PMID- 2986435 TI - Polyembryoma of testis. PMID- 2986436 TI - Ossified retroperitoneal fibrous histiocytoma. A case report. PMID- 2986437 TI - [Introduction to cerebral and spinal evoked potentials of genitourinary origin]. PMID- 2986438 TI - Macrophage functions in aging: effects of vitamin C deficiency. AB - Groups of young adult and senescent guinea pigs were fed normal and vitamin C deficient diets for 4 weeks and tested for their peritoneal macrophage functions. Serum levels of vitamin C in deficient animals indicated a progressive state of ascorbic acid deficiency and correlated well with the clinical signs and symptoms of scurvy. Fewer macrophages were obtained from the peritoneal cavities of deficient animals and morphologically they were smaller in size. Adverse effects of vitamin C deficiency were enhanced in aged animals. Significantly greater number of aged animals died by 4 weeks of deficiency. Deficient senescent animals had greater decline in macrophage random migration and bactericidal capacity. Following phagocytic stimuli, superoxide anion generation also significantly decreased. Data suggest that vitamin C deficiency might affect macrophage functions in the aged more profoundly and could compromise parameters of host defenses effective against microbial infections. PMID- 2986439 TI - Nuclear cardiology. AB - Radionuclide cardiac imaging is a safe, noninvasive alternative to cardiac catheterization for observation and evaluation of cardiac wall motion and calculation of ejection fraction. Nuclide imaging offers a greater degree of sensitivity and specificity in detecting myocardial ischemia and infarction than do conventional electrocardiographic and cardiac enzyme studies. It is especially useful in problem cases. Myocardial infarction can usually be evaluated with respect to size and relative age of infarction. PMID- 2986440 TI - Clinical pharmacology and therapeutic applications of the new oral converting enzyme inhibitor, enalapril. PMID- 2986441 TI - Variability and bias in the analyses of industrial hygiene samples. AB - Information on variability and bias of analyses of industrial hygiene samples is needed for quality control, for design of sampling programs, and for interpretation of results. Data was assembled from interlaboratory programs conducted by PAT-NIOSH, CDC, West Allis Memorial Hospital, and the Toxicology Center of Quebec for 12 types of samples. At the geometric mean concentrations of the reference samples the relative standard deviations of reported results were as follows: Pb, Cd, Zn on membrane filters--4.7%; solvents on charcoal tubes- 6.5%; Pb in blood--(CDC) 19%, (West Allis) 13%, (Quebec) 10%; Cd in blood--52%; Cd in urine--40%; As in urine--30%; Hg in urine--25%; F in urine--13%. For free silica on membrane filters, at the geometric mean value the geometric standard deviation was 1.29. For asbestos on membrane filters the statistical analysis was made on the square roots of the values, and the range of one standard deviation was from 68% to 138% of the average value. Four methods for Pb in blood were in close agreement, but three for As in urine differed substantially. Most methods did not achieve the reproducibility required by the NIOSH criteria. The probability of false positive and negative interpretation of results from each group of participating laboratories may be calculated from these data. PMID- 2986442 TI - Detection of chrysotile asbestos in workers' urine. AB - Urinary asbestos concentrations were evaluated as an indicator of occupational exposure to chrysotile asbestos via inhalation and ingestion. Detection of asbestos in the urine represents the first step in developing a biological indicator of exposure. Such an indicator could be used to supplement exposure data from workplace air sampling. A biological indicator would be particularly valuable in evaluating workers with intermittent airborne asbestos exposures and in determining if airborne exposure results in penetration of asbestos through the lung or gastro-intestinal tract. Transmission electron microscopy was selected as the most sensitive technique for identification of all sizes of asbestos fibers which might appear in the urine. First morning void urine samples were obtained from six workers (occupationally exposed to chrysotile asbestos in a factory producing roof coatings) and from a control group (six individuals with no occupational exposure). The levels of chrysotile asbestos detected in the urine of five workers were significantly greater than the asbestos concentrations in matched field blanks (both on a number and mass basis). Field blanks were designed to detect asbestos in the urine samples due to contamination which might occur during urine collection. Also, the workers' urinary asbestos levels were significantly greater than the concentrations found in the control group (both on a number and mass basis). Finally, the levels of chrysotile asbestos detected in the urine of two of six controls were significantly greater than those in matched field blanks (both on a number and mass basis). Although the project was not specifically designed to correlate urinary and airborne asbestos concentrations, preliminary data indicated that a correlation did not exist between these factors. PMID- 2986443 TI - Circulatory response to beta-adrenergic blockade at rest and during exercise. AB - The beta adrenoceptors involved in the regulation of the cardiovascular system include the beta 1 subtype in the heart, coronary arteries and juxtaglomerular cells of kidney, and the beta 2 subtype in skeletal muscle resistance vessels and on terminals of sympathetic nerves. The beta 1 receptors are activated primarily by norepinephrine released from the sympathetic nerves, the beta 2 by circulating epinephrine from the adrenal medulla. The function of these receptors is to adjust the circulation to meet the stresses imposed by gravitational forces including those that occur in changing from supine to standing position, in muscular exercise and during emotional stress. In normal subjects, other systems can compensate if the beta receptors are prevented from functioning. Thus, during beta-adrenergic blockade it is only when the cardiovascular system is taxed severely that deficiencies in its performance become apparent. In patients with cardiovascular diseases, other effects of beta blockers, not yet understood, may also be important. PMID- 2986444 TI - Long-term ingestion of guar gum is not toxic in patients with noninsulin dependent diabetes mellitus. AB - The use of diets rich in unabsorbable carbohydrate ("fiber") has been advocated for the treatment of noninsulin-dependent diabetes mellitus (NIDDM). The soluble viscous fibers such as guar gum are most effective in normalizing carbohydrate intolerance in such patients; particulate fibers such as cellulose have little or no effect. While the latter are known to affect many aspects of nutrition when consumed in great quantity, little is known of the toxicity of guar gum. Eight adults with NIDDM are reported here who consumed at least 30 grams of guar gum for at least 16 weeks without any change in hematologic, hepatic, or renal function. Serologic screening revealed no change in lipid, protein or mineral metabolism, and no change in electrolyte balance. It is concluded that consumption of 30 grams of guar gum per day for prolonged periods is without serious consequences. PMID- 2986445 TI - Stimulation of intestinal cytokinetics and mucin turnover in rats fed wheat bran or cellulose. AB - Rats were fed defined diets containing no fiber, 10% wheat bran or 10% cellulose, and intestinal morphology and cytokinetics were assessed by light microscopy and autoradiography, respectively. In bran-fed animals, there were no differences in morphological appearance of the jejunum, in the number of cells/villus column or in numbers of goblet cells compared to controls. Autoradiographic analysis, at one and 24 h after [3H]thymidine, however, suggested an increased turnover and villus transit of intestinal cells. There was also a 2.5 fold increase in incorporation of labeled sulfate, and a 2-fold increase in [3H]glucose incorporation into total intestinal glycoproteins and mucins. Similar, albeit less dramatic results were obtained in rats fed diets containing cellulose. These studies provide evidence that diets containing certain fiber derivatives can alter aspects of intestinal cell turnover, and support the earlier morphological observations suggesting increased goblet cell secretory activity in response to feeding these fiber derivatives. PMID- 2986446 TI - Mineral contents of brans passed through the human GI tract. AB - Concentrations of Cu, Fe, Zn, Ca, and phytate were measured in dry-milled corn bran, wheat brans, and soybean hulls prior to being baked in bread and after passage through the human GI tract. Significant changes in mineral concentrations in the retrieved remnants compared to the starting materials were as follows: Cu, Fe, and Zn increased by factors of from two to four and Ca increased at least ten fold in dry milled corn bran; Ca increased and phytate decreased in wheat brans; Zn increased and Fe decreased in soy hulls. These studies show that indigestible remnants of wheat brans in the human colon associate preferentially with calcium and that dry milled corn bran remnants can be loaded with increased concentrations of all four minerals, and especially with calcium. Concentrations of minerals and phytate were significantly greater in whole fecal samples from wheat bran diets than in the corresponding retrieved bran remnants. PMID- 2986447 TI - Influence of frequent and long-term bean consumption on colonic function and fermentation. AB - The objective of this study was to determine the influence of frequent and long term consumption of legume seeds on colonic function. Two groups of subjects were studied--one group habitually consumed legume seeds as part of their normal diet, a second group only infrequently consumed legumes. No differences between these groups could be detected for fecal output and frequency, intestinal transit time, VFA excretion or fecal pH during 23-day study periods in which subjects consumed either their usual diet or 100 g red kidney beans, daily. However, the addition of beans to the diets of both groups provided significantly more dietary fiber, and produced greater fecal output and a higher concentration of VFA in feces. Fecal output appeared to be determined by two independent parameters--dietary fiber intake and VFA excretion. Beans provided a physiologically useful source of dietary fiber and favorably influenced colonic function. PMID- 2986448 TI - Health implications of wheat fiber. PMID- 2986449 TI - Dietary changes and their possible effect on blood pressure. AB - In this summary, the authors have attempted to examine reports of associations between various dietary habits and practices on one hand, and serum lipids or clinical disease on the other hand. There seems to be little doubt that both hypertension and ischemic heart disease have a nutritional background, but in all likelihood, there are other factors such as hereditary traits, occupational hazards, and perhaps personal habits including cigarette smoking, alcohol abuse and prolonged ingestion of medicinal drugs. One of the strongest correlates seems to be the role of complex carbohydrates in regulating blood lipid concentrations. Carbohydrates not only have an effect on the endocrine system that regulates blood volume, but they also influence absorption of fat soluble substances from the digestive tract and if natural fiber is included, it has an effect on fecal bulk, transit time of the fecal stream, and reabsorption of bile acids and neutral sterols. Epidemiologically, there is some evidence that the changes that occurred in the American diet in the years between 1914-1944 may well have played a permissive role in the genesis of a portion of the coronary heart disease, high blood pressure and stroke that occurred in the United States. It is not too farfetched to suggest that had the American servicemen been given more cereal food products including bread and other baked food items, instead of excessive amounts of meat and fats, the dietary pattern of America might well have been substantially different. Furthermore, this difference could easily have influenced the pattern of atherosclerosis and hypertension. A great deal more work is needed to confirm or refute these suggestions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986450 TI - Clinical and pathologic findings of the liver in the acquired immune deficiency syndrome (AIDS). AB - Clinical data and histologic sections of the liver, including immunohistochemical studies for hepatitis B surface and core antigens, were reviewed in 42 autopsy cases of the acquired immune deficiency syndrome (AIDS). Hepatomegaly, elevation of serum transaminases, and mild elevation of alkaline phosphatase were commonly observed clinical and biochemical abnormalities. Mildly elevated alkaline phosphatase and normal bilirubin levels were present in patients with Mycobacterium avium-intracellulare (MAI) infection, cytomegalovirus (CMV) infection, and Kaposi's sarcoma (KS). Histologic sections demonstrated liver involvement by MAI in eight cases; KS in six cases; cryptococcus in three cases; and CMV in two cases. One case of MAI infection was associated with marked central vein sclerosis, a finding previously unreported. Thirty-two (76%) of 42 cases had serologic or pathologic evidence of hepatitis exposure. Two patients had histologic evidence of chronic active hepatitis. The pathologic processes involving the liver appeared to be secondary to the infections and neoplasms for which this population is susceptible and did not significantly contribute to morbidity or mortality. No findings specific or pathognomic for AIDS were identified in the liver. PMID- 2986451 TI - Reactivity of serologic tests for the detection of antibody specific to cytomegalovirus. AB - A total of 212 sera were assayed for antibody specific for cytomegalovirus by complement fixation (CF), indirect immunofluorescence (IFA Electro-Nucleonics Laboratory, Inc., Bethesda, MD), ELISA (Cordis Laboratories, Inc., Miami, FL) and the FIAX system (International Diagnostic Technology, Santa Clara, CA). Correlation of CF with IFA, ELISA, and FIAX was 61%, 78%, and 71%, respectively. Quantitative correlation between IFA and FIAX and ELISA was not possible because of the broad range of reaction intensity of the latter two tests in sera with a particular IFA titer. PMID- 2986453 TI - Partial splenic embolization as an emergency treatment for uncontrollable variceal bleeding: a case report. AB - Partial splenic artery embolization was carried out in one patient with cirrhosis during acute variceal bleeding when other direct hemostatic procedures could not be applied. It clearly assisted in controlling the acute bleeding, and led to subsequent endoscopic sclerosis. Although its effect is indirect, partial splenic embolization may prove useful in certain clinical situations with acute bleeding where other hemostatic means have failed or are not applicable. PMID- 2986452 TI - Colonic bacterial flora and serum cholesterol: alterations induced by dietary citrus pectin. AB - The influence of dietary pectin on blood lipids and stool bacterial flora, enzyme activity, and physical properties was investigated. Fifteen grams of citrus pectin were added to the normal diets of 10 subjects. A decrease in serum cholesterol was observed after 2 wk of dietary supplementation, but was not statistically significant after the 3rd wk. Fecal bacterial flora was modified by pectin; anaerobic bacteria increased and aerobic bacteria increased. No change in stool bacterial 7 alpha dehydroxylase or cholesterol dehydrogenase activity was observed. The daily stool output, pH, and water content were not influenced by the addition of pectin to the diet. PMID- 2986454 TI - The role of renal catecholamines in hypertension. AB - We have found increased renal alpha 2-adrenoceptor density and a defect in prostaglandin and parathyroid stimulated adenylate cyclase in two genetic forms of rat hypertension. Changes in serum calcium and parathyroid hormone (PTH) levels suggest biologic significance to this defective adenylate cyclase response. Our hypothesis is that one or more of these defects contribute to excess renal retention of sodium and increase vascular resistance of genetically hypertensive rats and humans with essential hypertension who have similar abnormalities of calcium and PTH. PMID- 2986455 TI - Silica, pneumoconiosis, and carcinoma of the lung. AB - The current argument about the carcinogenicity of inhaled silica is not clarified by reliance on morbidity and mortality experience divorced from or incompletely related to data on environmental exposure. Human evidence provides the ultimate basis for assessing such risks, and numerous studies of the effects of inhaling dusts rich or poor in silica content on the prevalence of pulmonary carcinoma have been performed on large series of cases from major mining areas of the world. When due allowance is made for substances inhaled concomitantly with exposure to silica and for personal pollution by cigarette smoking, the weight of evidence is against a carcinogenic role for uncombined silicon dioxide. Moreover, pneumoconiosis due to compact mineral particles does not appear to determine the onset of lung cancer. Cellular behaviour suggests reasons for the different responses to compact and fibrous particles acting alone. PMID- 2986456 TI - Genetics and atomic bombs in Hiroshima and Nagasaki. PMID- 2986457 TI - A previously undescribed autosomal recessive multiple congenital anomalies/mental retardation (MCA/MR) syndrome with fronto-nasal dysostosis, cleft lip/palate, limb hypoplasia, and postaxial poly-syndactyly: acro-fronto-facio-nasal dysostosis syndrome. AB - We describe two sibs born to a consanguineous couple. Among other clinical findings both have mental retardation, short stature, facial and skeletal abnormalities characterized by hypertelorism, broad notched nasal tip, cleft lip/palate, campto-brachy-poly-syndactyly, fibular hypoplasia, and marked anomalies of foot structures. Facial signs of the reported patients resemble those present in the fronto-nasal "dysplasia" syndrome; however, the whole clinical picture in the present patients suggests a true MCA/MR syndrome, most likely inherited as an autosomal recessive trait. Clinical and genetic aspects of the present family are discussed. PMID- 2986458 TI - Methotrexate with citrovorum factor rescue in gestational trophoblastic disease. AB - Thirty-three patients with gestational trophoblastic disease treated with methotrexate alone and 68 similar patients treated with methotrexate and citrovorum factor rescue were analyzed. Both groups showed a comparable sustained biochemical remission rate. The group treated with methotrexate and citrovorum factor however showed a shorter interval to induction of remission (p less than 0.05). Contrary to previous reports, the methotrexate and citrovorum factor regimen failed to protect the patients against the development of hepatic toxicity, the incidence of hepatic toxicity being significantly higher in the group treated with methotrexate and citrovorum factor. PMID- 2986459 TI - Orthodontic therapy in patients with juvenile periodontitis: clinical and microbiologic effects. AB - The correction of malocclusions in juvenile periodontitis (JP) patients completing periodontal therapy is a problem of increasing clinical concern to orthodontists, since many teeth with severe alveolar bone loss in these patients can now be successfully treated without extraction. In this report, fixed edgewise orthodontic therapy was carried out after the completion of periodontal therapy on four JP patients. The orthodontic therapy included extensive intrusion of teeth severely affected by JP. Phase-contrast microscopic analysis of subgingival plaque from orthodontically treated teeth was used to monitor longitudinally the effects of fixed orthodontic bands on the subgingival flora and also to monitor the efficacy of topical and systemic antimicrobial therapy aimed at suppression of suspected periodontopathic bacteria. Orthodontic movement was completed on most periodontally compromised teeth without significant evidence of additional deterioration in periodontal status. However, within the first 6 months of orthodontic band placement, all patients had significant increases in the number of spirochetes and motile rods in their subgingival flora. Three of the patients also developed high levels of crevicular polymorphonuclear leukocytes around orthodontically treated teeth, indicating significant subgingival inflammation. Intensive antimicrobial measures, including topical inorganic salt applications and systemic tetracycline, were helpful in limiting clinical inflammation and subgingival colonization by periodontopathogens during orthodontic therapy. The results demonstrate that successful orthodontic repositioning can be carried out in treated JP patients. In addition, bacteriologic monitoring and chemotherapeutic suppression of periodontal pathogens may be valuable in the prevention of further destructive periodontal disease activity in periodontitis patients undergoing orthodontic therapy. PMID- 2986460 TI - Evidence for the origin of Kaposi's sarcoma from lymphatic endothelium. AB - Previous studies utilizing enzyme histochemistry, electron microscopy, and immunohistochemistry have failed to establish the cell of origin in Kaposi's sarcoma. The authors have rigorously tested the prevailing hypothesis that the lesion defined as Kaposi's sarcoma is derived from vascular endothelial cells. They use seven markers to characterize endothelial cells: three antigens (Factor VIII-related antigen, HLA-DR/Ia, macrophage/endothelial antigens), three enzymes (5'-nucleotidase, ATPase, alkaline phosphatase), and lectin binding (Ulex europaeus I). They applied the markers first to normal skin and lymph node, and then to biopsy specimens from 40 patients with Kaposi's sarcoma. Normal blood vessel endothelium was positive for all seven markers, but normal lymphatic endothelium was negative for all of the markers except 5'-nucleotidase and Ulex europaeus lectin. The neoplastic cells in 40 cases of Kaposi's sarcoma closely resembled those of normal lymphatic endothelium but not those of blood vessel endothelium. This suggests that Kaposi's sarcoma may originate in lymphatic endothelium. PMID- 2986461 TI - Sodium-pump density of cells from dog tracheal mucosa. AB - Uptake of tritiated ouabain by cells isolated from dog tracheal epithelium showed two components: a saturable component with a Km of 5.1 X 10(-8) M and a maximal uptake of 8.3 X 10(5) molecules/cell and a nonsaturating component of uptake that was linear with concentration. Several criteria indicated that the saturable uptake component represented binding to the Na+-K+-ATPase. To estimate the average surface area per cell, a known number of cells were pelleted and weighed, and the average surface area was calculated, assuming the cells to be perfectly spherical. The validity of this assumption was confirmed by comparing the calculated surface areas of cells in isotonic and hypotonic media. From the values for maximal saturable uptake and average surface area, a pump density of approximately 2,400 sites/micron2 was calculated. Given that the apical membrane lacks Na pumps and accounts for only approximately 5% of the total surface area, this value corresponds to the pump density of the basolateral cell membrane. The pump densities of ciliated, goblet, and basal cells were compared by autoradiography. The three cell types had approximately the same density of pump sites. PMID- 2986462 TI - Epithelial properties of human colonic carcinoma cell line Caco-2: effect of secretagogues. AB - Human colonic carcinoma Caco-2 cells grown in vitro form epithelial layers of highly polarized cells. Unlike colonic adsorptive cells they possess a mucosal membrane with very limited ionic conductance, even after exposure to aldosterone. When grown on filters, Caco-2 cells were sensitive to various secretagogues; these included 10(-5) M dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) and 10(-10) M vasoactive intestinal peptide, both of which, added serosally, enhanced the short-circuit current. The same applied to mucosal forskolin. Caco-2 cell sensitivity to serosal epinephrine was lower. Ion substitutions and 22Na 36Cl flux measurements indicated the possibility of secretagogue-dependent chloride secretion. Measurements on cells grown on Petri dishes and exposed to 1 mM DBcAMP for 1 h enabled detection of more profound modifications. Sustained 20 mV cell depolarization and a large reduction in the relative electrical resistance of the mucosal membrane were concomitant with a sizable decrease in 36Cl accumulation. These results suggest that Caco-2 cells, which to some extent resemble colonic crypt cells, possess the cAMP-dependent mucosal chloride conductance characteristic of secretory cells. PMID- 2986463 TI - Cultured rat vascular smooth muscle cells: extracellular calcium and Na+-K+ regulation. AB - This study explores the relationship between extracellular calcium (Cao) and Na+ K+ regulation as it particularly pertains to the activity of the Na+ pump in cultured vascular smooth muscle cells (VSMCs) originating from Sprague-Dawley rats. As compared with cells incubated in media containing 0.5, 2.0, or 4.0 mM calcium, when the Na pump is active, VSMCs incubated in a Ca-deficient medium show a marked increase in intracellular sodium and no significant change in intracellular potassium. Associated with the rise in intracellular sodium there is an augmented activity of the Na pump. When the Na pump is inhibited, VSMCs incubated in either high-Ca medium (Cao = 4.0 mM) or Ca-deficient medium manifest a greater decline in intracellular potassium than cells incubated in media containing 0.5 or 2.0 mM calcium. Furthermore, when the Na pump is inhibited, VSMCs incubated in a Ca-deficient medium exhibit higher intracellular sodium levels in comparison with their counterparts incubated in media containing calcium. Flux experiments indicate that the aforementioned changes reflect increased membrane permeabilities to Na+ and K+. It is concluded that by regulating the permeability of the VSMC membrane, Cao plays an important role in the intracellular Na+-K+ homeostasis and that its effect on the Na pump is mediated via perturbations in the intracellular Na+ and K+ concentrations. PMID- 2986464 TI - Effects of tumor promoters on sodium ion transport across frog skin. AB - Phorbol esters are tumor promoters and mitogens whose effects may be mediated by changes in ion transport across membranes. Clarification of the transport effects of these agents should be facilitated by using a well-characterized model epithelial system whose intracellular and transmural parameters are readily measurable. The current results constitute a preliminary study of the effects of 12-O-tetradecanoylphorbol-13-acetate (TPA), phorbol-12,13-dibutyrate (PDBU), and phorbol on the short-circuit current (Isc) across frog skin. TPA produced two effects: a stimulation of Isc of variable magnitude and a far more constant inhibition of the natriferic action of vasopressin. These effects appear related to the action of TPA as a tumor promoter insofar as PDBU (an active ester) also inhibited the natriferic response to vasopressin, whereas phorbol (inactive as a tumor promoter) had no significant effect. TPA is largely active from the mucosal medium, inhibits the natriferic response to adenosine 3',5'-cyclic monophosphate (cAMP) as well as that to vasopressin, and does not stimulate Isc in the presence of 10(-4) M mucosal amiloride. Inhibition of prostaglandin E1 production by indomethacin had no effect on the actions of TPA. The results indicate that frog skin is a promising model for studying the transport effects of the phorbol esters. The data further suggest that TPA acts on frog skin by activating the physiological amiloride- and cAMP-sensitive channels gating apical Na+ entry from the mucosal medium into the epithelial cells. PMID- 2986465 TI - cAMP-independent stimulation of glycogen phosphorylase in newborn rat hepatocytes. AB - Postnatal development of glycogen phosphorylase activation by the cAMP independent pathway was examined in isolated rat hepatocytes from control and propylthiouracil-treated congenital hypothyroid rat pups. At 5 days postnatum there was complete phosphorylase activation by beta-adrenergic stimulation, glucagon, and the calcium ionophore A23187, but no activation by alpha-adrenergic stimulation. Activation of phosphorylase by angiotensin or vasopressin was less than in hepatocytes from adult rats (P less than 0.01). At 28 days postnatum activation by all of these hormones was complete. In the propylthiouracil-treated group hormone responsiveness was similar to the control at 5 days postnatum. However, alpha-adrenergic (P less than 0.025), angiotensin, and vasopressin (P less than 0.05) activation was decreased at 28 days postnatum, and beta adrenergic, glucagon, and A23187 activation was complete. The attenuated responses were restored by thyroxine replacement from 15 days postnatum. [32P]Pi incorporation into phosphatidylinositol by epinephrine and vasopressin in 28-day propylthiouracil-treated rats was lower than the control (P less than 0.01). We speculate that the diminished phosphorylase response of hepatocytes to alpha adrenergic, vasopressin, or angiotensin stimuli in the early neonatal period could be related to low receptor numbers and the weaker phosphoinositide response during this period. Also, the depressed phosphorylase response to alpha adrenergic, vasopressin, and angiotensin stimulation in congenital hypothyroidism at 28 days postnatum could be related to a decrease in number of plasma membrane receptors for these agonists. PMID- 2986466 TI - Gastric vasoconstrictor actions of leukotriene C4, PGF2 alpha, and thromboxane mimetic U-46619 on rat submucosal microcirculation in vivo. AB - The gastric vasoconstrictor actions of the arachidonate lipoxygenase products leukotrienes B4, C4, and D4 and the prostanoids prostaglandin F2 alpha (PGF2 alpha) and the endoperoxide analogue U-46619 have been investigated in vivo in the submucosal microcirculation of the anesthetized rat using direct microscopy. Topical application of PGF2 alpha (1-100 microM) to the exposed submucosa reduced vessel diameter of the venules, with peak vasoconstriction occurring within 1 min and remaining during the 3-min period of administration. Constriction of the arterioles by PGF2 alpha was less pronounced. U-46619 (1-1,000 nM), a thromboxane mimetic, induced vasoconstriction in both arterioles and venules, reaching plateau responses within 1.5-2.0 min of application. Leukotriene C4 (25-400 nM) induced vasoconstriction in the venules, which was more pronounced than in the arterioles, reaching peak responses within 1-1.5 min, which were unaffected by indomethacin administration. No significant vasoactive action with leukotrienes B4 or D4 in the submucosal microcirculation could be detected. With both leukotriene C4 and U-46619 intense focal vasoconstriction in the venules was observed, leading to sluggish blood flow or stasis within the vessel. In contrast, norepinephrine had no significant action on submucosal venules at concentrations that substantially reduced arteriolar vessel diameter. Such potent vasoconstrictor actions of leukotriene C4 and U-46619 in the gastric submucosa identify this leukotriene and thromboxane A2 as potential endogenous proulcerogenic agents and suggest that these arachidonate products could play a role in microcirculatory events accompanying gastric damage. PMID- 2986467 TI - Interaction of left atrial receptors and carotid sinus baroreceptors on heart rate in the dog. AB - This study was undertaken to determine the influence of 1) the left atrial receptors (LA) on the ability of the carotid sinus baroreceptors (CS) to regulate heart rate and 2) the CS on the reflex increase in heart rate mediated by the LA. The LA were stimulated by stretching the pulmonary vein-atrial junctions in dogs anesthetized with alpha-chloralose. Aortic pressure was controlled, and the pressure in the CS was regulated. Stimulus-response curves were obtained relating heart rate to pressures in the CS, in the control state, and during stimulation of LA (6 dogs). Factorial analysis revealed that LA exerted a significant influence on heart rate (P less than 0.01). Next the CS pressure was set at mid, low, and high levels and the LA stimulated. It was found that the effect on heart rate was greatest at the mid setting (+19.3 +/- 2.9 beats/min) and least at the low setting of the pressure in the CS (+0.9 +/- 0.5, 11 dogs). Sympathetic blockade attenuated significantly the response in the mid setting of the CS pressure and left intact the response at the high setting of the pressure in the CS. It is concluded that there is a significant interaction between these two reflexes. PMID- 2986468 TI - Beta-adrenergic modulation of direct defibrillation energy in anesthetized dog heart. AB - Catecholamines facilitate ventricular defibrillation in animals. We examined the effects of beta-adrenergic stimulation and blockade on ventricular defibrillation threshold in anesthetized dogs. Calibrated shocks were delivered between epicardial and superior vena caval electrodes, and defibrillation threshold was measured before and after administration of isoproterenol and propranolol. Eight dogs (group 1) received isoproterenol before propranolol. Nine dogs (group 2) received propranolol before isoproterenol. In group 1, the minimum energy required to defibrillate before isoproterenol was 10.6 +/- 1.7 (SE) J and decreased to 5.9 +/- 1.3 with isoproterenol (P less than 0.001). In group 2, the minimum energy required to defibrillate was 8.3 +/- 2.4 J before propranolol and increased to 10.7 +/- 2.2 after propranolol (P less than 0.001). In group 1, propranolol after isoproterenol increased defibrillation threshold (P less than 0.07), whereas in group 2 isoproterenol after propranolol produced no significant change in defibrillation threshold. Thus beta-stimulation decreased defibrillation threshold significantly in the anesthetized dog heart, an effect that was blocked by propranolol. Conversely, propranolol increased defibrillation threshold, an effect that occurred despite prior beta-stimulation, probably because of the short half-life of isoproterenol. PMID- 2986469 TI - Potassium ion homeostasis and mitochondrial redox status of turtle brain during and after ischemia. AB - To account for the remarkable capacity of turtle brain to survive without oxygen, microelectrode recordings of extracellular potassium activity (K+o) were made in intact brains of normoxic pentobarbital-anesthetized turtles during and after ischemia. Changes in redox status of brain cytochrome a,a3 were recorded simultaneously by reflection spectrophotometry. Cytochrome a,a3 became fully reduced, and K+o rose at a slow rate for approximately 60 min during complete ischemia. When K+o reached 8-12 mM, an inflection point was observed after which K+o rapidly increased to levels above 30 mM. Full recovery of baseline K+o and cytochrome a,a3 redox state occurred only if brain blood flow was restored within 60 min. These data demonstrate that maintenance of ion homeostasis in turtle brain depends on blood flow and continued delivery of systemic substrates and that viability is threatened if ischemia is prolonged beyond 60 min. PMID- 2986470 TI - Breast carcinoma presenting with axillary lymph node metastases. An analysis of specific histopathologic features. AB - Forty-three patients with metastatic carcinoma in axillary lymph nodes from an unknown primary site were studied. Thirty-one women (72%) were subsequently identified as having primary carcinoma in the ipsilateral breast. Based on the histologic patterns of the metastasis, patients were divided into three groups. Type I axillary lymph node metastases were composed of sheets of large, apocrine like pleomorphic cells with pale to granular pink cytoplasm, large nuclei, and prominent nucleoli. This type was found in two-thirds of the cases. Type II metastases were readily recognizable as characteristic of breast carcinoma and included glandular, cribriform, and papillary patterns and comedonecrosis. Type III metastases demonstrated a mixture of the two previous patterns. Patients proven to have breast carcinoma ranged in age from 33 to 83 (average 58). Among the 31 cases with proven breast carcinoma, mammography was abnormal in 11 (35%). In the 12 other cases (18%), no primary in the breast or other site was demonstrated. Patients not proven to have breast carcinoma had a similar age distribution, comparable proportions of histologic patterns of metastases, and similar survival results to those with a demonstrated breast carcinoma, and none of the 12 patients were later shown to have another primary. The findings described here indicate that when mammary carcinoma presents initially with axillary metastases, it often has a distinctive histological pattern. In most cases this consists of relatively large, apocrine-like cells growing diffusely without forming glands or papillary structures. A minority of metastases have patterns (comedonecrosis; trabecular, glandular, or trabecular growth) more characteristic of the usual spectrum of breast carcinoma. Awareness of this morphological diversity should assist the pathologist in suggesting the breast as a primary site when the initial manifestation is an axillary lymph node metastasis. PMID- 2986472 TI - Hepatocellular carcinoma. A 5 year institutional experience. AB - Liver cancer is the most common of all malignancies worldwide, its incidence reaching almost epidemic proportions in some countries. However, its significance in North America has generally been underemphasized. In a 5 year period, hepatocellular carcinoma was diagnosed in 35 adult patients in our institution. Preexisting liver inflammation was present in 27 of the patients (77 percent). Although most patients had symptoms, they included only poorly defined pain or cachexia in most cases, and 10 patients (29 percent) were totally asymptomatic at the time of diagnosis. Five patients presented with hemoperitoneum due to intraabdominal tumor rupture. Examinations useful in confirming the diagnosis included alpha-fetoprotein determination, liver scan, and computerized tomographic scanning. Eight patients (23 percent) had associated visceral or other malignancies. The outlook was poor, with a mean survival of 5 months, and only two patients survived more than 1 year. Hepatocellular carcinoma is uncommon but not at all rare in the Pacific Northwest. It arises from chronic liver inflammation, is diagnosed late, and has a grim prognosis. Prevention of various forms of chronic liver inflammation, and mass serial screening of populations at high risk for the development of hepatocellular malignancy will probably have the greatest role in reducing the lethality of this disease. PMID- 2986471 TI - Immature teratoma of the ovary with predominant malignant retinal anlage component. A parthenogenically derived tumor. AB - An immature teratoma with predominant malignant retinal anlage component occurred in the ovary of a 17-year-old woman. Karyotype analysis from Q-banded preparations revealed that the tumor was parthenogenically derived and exhibited chromosome instability and aneuploidy. On light-microscopical examination, the malignant elements resembled retinal anlage. Ultrastructurally, large, polygonal, pigmented cells had features of ependymal epithelium: a continuous, well developed basal lamina; microvilli; parallel arrays of rough endoplasmic reticulum; and very large, densely pigmented melanosomes. Adjacent cells identified by light microscopy as immature neuroglia contained glial fibrillary acidic protein by immunohistochemical staining. The presence of neuroglia, not a feature of tumors of neural crest origin, and the ependymal characteristics of the pigmented cells indicate that the neoplasm was most likely of neural groove derivation. PMID- 2986473 TI - Dendritic inclusions in the cerebellar granular layer after long term alcohol consumption in adult rats. AB - Whorled multiplanal inclusions up to 1.5 micron in diameter were observed in dendrites of the cerebellar Golgi cells after 6 months of alcohol treatment, increasing in number with time. Inclusions are formed by apposed cytomembranes stacked parallel or in a twisted arrangement. The paired membranes merged into a dense, finely textured material. A close relationship with dendritic smooth endoplasmic reticulum is apparent. A correlation between chronic alcohol intake and the development of these inclusions is shown although, in common with many other types of neuronal cytoplasmic inclusions, their genesis and function remains to be determined. PMID- 2986474 TI - The effect of magnesium trisilicate mixture, metoclopramide and ranitidine on gastric pH, volume and serum gastrin. AB - Eighty women (40 for elective Caesarean section and 40 for elective gynaecological surgery) were randomly assigned to one of five treatment groups and received pre-operatively either no medication; magnesium trisilicate mixture (BP) 30 ml; metoclopramide 10 mg intramuscularly; ranitidine 150 mg orally on the night prior to, and the morning of, surgery; or metoclopramide 10 mg intramuscularly in combination with oral ranitidine 150 mg (the latter again given on the night prior, and the morning, of surgery). The effect of these medications on intragastric pH, volume and serum gastrin-17 was measured. No patient receiving ranitidine had a pH of less than 4. Magnesium trisilicate mixture resulted in the largest intragastric pH change although one woman in this group had a pH of 1.7. The largest intragastric volumes were seen in the patients who had received magnesium trisilicate mixture, whilst the patients who had received metoclopramide in combination with ranitidine had the smallest intragastric volumes. Magnesium trisilicate mixture and metoclopramide resulted in no change in serum gastrin levels. However, in the subjects who had received ranitidine on the night prior to surgery, the fasting serum gastrin was significantly higher (p less than 0.01) than the values in the remaining subjects, the mean (SEM) values being 60.3 (6.3) pg/ml in those not receiving ranitidine and 111.3 (19.5) pg/ml in those who had been given ranitidine. PMID- 2986476 TI - A hydroxylapatite microassay for receptor binding of 2,3,7,8-tetrachlorodibenzo-p dioxin and 3-methylcholanthrene in various target tissues. AB - A "batch" hydroxylapatite assay for the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) receptor that does not require detergents is described. The receptor could be assayed in rat target tissues using either of the cytochrome P1-450 inducers [3H]TCDD or [3H]3-methylcholanthrene as radioligands. A phosphate buffer washing procedure was developed on the basis of chromatographic data and optimized to separate nonspecifically and specifically bound ligand. The assay was characterized with respect to washing efficiency, binding specificity, competition, adsorption time, amount of hydroxylapatite required to bind receptor complexes, sensitivity, and effects of detergents. Equilibrium binding parameters were determined. Receptor extracted with phosphate from hydroxylapatite was analyzed on sucrose gradients and was found to exhibit the same sedimentation properties as the receptor in crude cytosol. Furthermore, the applicability of the assay has been demonstrated in cytosolic preparations from three different target tissues: liver, lung, and thymus. PMID- 2986477 TI - Quantitation of chemically induced DNA strand breaks in human cells via an alkaline unwinding assay. AB - DNA strand breaks induced in human CCRF-CEM cells by electrophilic chemicals (carcinogens/mutagens) can be readily quantitated via a facile alkaline unwinding assay. This procedure estimates the number of chemically induced DNA strand breaks on the basis of the percentage DNA converted from double-stranded to single-stranded form during an exposure to the alkaline unwinding conditions. The assay is based on the assumption that each strand break serves as a strand unwinding point during the alkaline denaturation. The extent of strand separation can be standardized with respect to the initial level of induced strand breaks by the use of X-rays, which produce known levels of DNA strand breaks per rad in mammalian cells. Subsequent to the alkaline exposure, the single- and double stranded DNA were separated by use of thermostated hydroxylapatite columns (60 degrees C), and the DNA was quantitated via a fluorescence assay (Hoechst 33258 compound). A correlation was shown between mammalian DNA strand-breaking potential (as measured in this procedure) and the propensity of these chemicals to revert Salmonella typhimurium TA100. PMID- 2986475 TI - Monoclonal antibodies in enzyme research: present and potential applications. AB - Hybridoma antibodies are powerful tools. Their impact is already apparent in many areas of basic and applied research. In contrast, their impact is just beginning to be felt in enzymology. The existing literature on monoclonal antibodies to enzymes and isozymes, reviewed in this article, is as yet largely descriptive. However, the potential applications discussed herein promise to revolutionize existing strategies of unraveling the basic biochemistry, immunochemistry, and developmental, somatic cell, and molecular genetics of enzymes and isozymes. At a clinical level, monoclonal antibodies to enzymes promise to radically improve the current modalities of diagnosis and therapy in clinical enzymology and oncology. It is becoming increasingly apparent that the future applications of hybridoma antibodies to enzymes and isozymes appear to be limited only by our imagination. PMID- 2986478 TI - Influences of salts on high-performance liquid chromatography of leukotrienes. AB - The effects of concentrations and kinds of salts on the resolution of leukotrienes (LT) C4, D4, E4, and B4 were investigated by two kinds of reversed phase high-performance liquid chromatography columns (muBondapak C18 and Novapak C18). When a mobile phase (acetonitrile/methanol/water) with a lower concentration of acetic acid (0.02-0.1%) at pH 5.6 was used, LTC4 and LTD4 were not eluted from the muBondapak column. On the Novapak column, LTC4 and LTD4 were eluted, but they were poorly resolved. When the concentration of acetic acid in the mobile phase was raised to 1.0% and adjusted to pH 5.6 with ammonium hydroxide or triethylamine, excellent resolution of LTs was obtained. Sodium hydroxide was, to some extent, useful for the pH adjustment of the mobile phase. Sodium chloride could not be substituted for acetic acid-ammonium hydroxide or triethylamine salt. The resolution of LTC4, LTD4, and LTE4 was affected more strongly than that of LTB4 by changes of concentrations and kinds of salts. When the acetonitrile/methanol/water/acetic acid solvent system adjusted to pH 5.6 with triethylamine was applied to the analysis of the leukotrienes produced from rat peritoneal cells with stimulation of calcium ionophore A23187, de novo synthesized LTC4, LTD4, LTB4, and isomers were clearly separated. This solvent system may be useful for the investigation of variations in the synthesis of subclasses of LTs with different stimuli and under different circumstances. PMID- 2986479 TI - Screening for amplification of specific DNA sequences in cultured mammalian cells. AB - Carcinogen- or herpes virus-induced amplification of integrated simian virus 40 DNA in transformed cell lines has been demonstrated previously by Southern blot analyses, dot hybridizations, and dispersed cell assays. Although these methods are quite reliable, large series of experiments require a considerable amount of time and effort. Therefore, an alternative method has been developed that allows one to carry out great numbers of separate gene-amplification experiments in the 96 wells of microtiter plates and to analyze the DNAs of the treated cells simultaneously after "replica blotting" onto membrane filters by hybridization with a cloned DNA probe. PMID- 2986480 TI - Purification of Gc-2 globulin from human serum by displacement chromatography: a model for the isolation of marker proteins identified by two-dimensional electrophoresis. AB - A protein that was initially known only as a minor spot in two-dimensional electrophoresis patterns of serum obtained from certain psoriasis patients, particularly those with a pustular component to their disease, has been purified by two stages of ion-exchange displacement chromatography on DEAE-Sephacel at different pH levels, followed by elution chromatography on hydroxylapatite. The purification was followed by examining the column fractions directly by two dimensional gel electrophoresis. The capacity of the displacement system, which utilized carboxymethyldextrans as displacers, was very high; 6 ml of dialyzed serum applied to a 7-ml column in the initial stage resulted in a very substantial enrichment of the target protein. The second displacement stage yielded a highly purified product, contaminated only by A-1 lipoprotein. The latter was removed by hydroxylapatite chromatography. The purified protein was subsequently identified as Gc-2 globulin, a vitamin D-binding protein, by immunological procedures. The results demonstrate the effectiveness of ion exchange displacement chromatography in focusing resolving power on the relatively narrow range of affinities represented by the target protein and its immediate neighbors in a chromatogram, as well as the applicability of the system to the isolation of a protein known only by its position in a two-dimensional electrophoretic pattern. PMID- 2986481 TI - An enzymatic inosine 5'-monophosphate assay of increased specificity. AB - The enzymatic inosine 5'-monophosphate assay described by Grassl [in, Methods of Enzymatic Analysis (H. U. Bergman, ed.), pp. 2168-2171, Academic Press, New York (1974)] is highly nonspecific, as ITP, ATP, ADP, AMP, and adenosine react stoichiometrically. The reactivity with the adenine derivatives is due to the tri and diphosphatase activity of alkaline phosphatase (AP), coupled with adenosine deaminase (and possibly AMP deaminase) contamination of commercially available preparations of AP, purine-nucleoside phosphorylase, and/or xanthine oxidase. The inclusion of coformycin (0.05 microgram/ml), a potent inhibitor of these deaminases, completely eliminated the cross-reactivity. ITP, however, still reacted stoichiometrically due to the tri- and diphosphatase activity of AP. Meyer and Terjung [Amer. J. Physiol. 237 C111-C118 (1979)] introduced a modification of Grassl's procedure, substituting 5'-nucleotidase for AP. It has been found that this disallows reactivity with ATP, ADP, and ITP but that AMP and adenosine still react completely. Coformycin prevents this cross-reactivity. It is therefore recommended that the assay be carried out with 5'-nucleotidase (instead of AP) and coformycin, in order to achieve a more specific assay, and one more suitable for use with whole tissue extracts. PMID- 2986482 TI - The basolateral membrane of rat enterocyte: its purification from brush border contamination. AB - Basolateral membranes obtained by self-orienting Percoll-gradient centrifugation were treated with 5 mM CaCl2 to minimize the cross-contamination by brush border membranes. From marker enzyme-specific activities it was calculated that in this preparation the basolateral/brush border membrane ratio was 22.6. A low L-glucose permeability across basolateral membrane vesicles together with ATP-dependent sodium uptake was observed. PMID- 2986483 TI - Percoll and Ficoll self-generated density gradients by low-speed osmocentrifugation. AB - Percoll and Ficoll self-generated density gradients can be obtained by low-speed centrifugation of their solutions within dialysis cells. Useful Percoll density gradients can be obtained after 10-30 min centrifugation at 220-2010g, within dialysis cells. Ficoll density gradients, which are more difficult to self generate, can be obtained by the same technique. Red cell band formation in a Percoll density gradient can be done in a single step by using dialysis cells as the centrifugation solution container. PMID- 2986484 TI - Chitin-coated Celite as an affinity adsorbent for high-performance liquid chromatography of lysozyme. AB - Preparation of chitin-coated Celite as an affinity adsorbent for high-performance liquid chromatography of lysozymes and its application to separation of N bromosuccinimide-oxidized lysozymes are described. By pH gradient elution, two diastereomers of oxindolealanine-62-lysozyme, delta 1-acetoxytryptophan-62 lysozyme (intermediate product in the reaction in acetate buffer), and native lysozyme were all separated within 40 min. PMID- 2986485 TI - Electrophoresis improves accuracy of paternity testing. PMID- 2986486 TI - Activation of actin-containing microfilaments by vasopressin in the amphibian urinary bladder epithelium: a fluorescent study using NBD-phallacidin. AB - The effect of an antidiuretic hormone (ADH, vasopressin) on the microfilament system of the toad urinary bladder lumenal epithelium was investigated using NBD phallacidin (NBD-ph). The latter material is a specific fluorescent label for F actin. In the presence of an osmotic gradient, both ADH and cyclic adenosine monophosphate (cAMP) appear to induce the polymerization of monomeric actin into F actin-containing microfilaments. The latter may then be involved in the morphological changes, including the formation of lateral intercellular lakes, associated with the typical hydroosmotic response. PMID- 2986487 TI - Androgen dependence of testicular and epididymal angiotensin converting enzyme. AB - Treatment with cadmium chloride (CdCl2) and cyproterone acetate (CA) depressed angiotensin converting enzyme (ACE) activity significantly in testes and epididymal regions of the adult rats compared to the corresponding untreated controls. Exogenous testosterone to CA-treated rats significantly increased the enzyme activity both in the testes and epididymis, the effect in the latter being very significant comparable to CA-treated and untreated controls. Testosterone failed to induce ACE activity in the testes and caput epididymis of 30 day-old immature rats, but the enzyme activity was detected in corpus and cauda epididymis. Our findings indicate that ACE activity in the testicular complex is possibly linked with androgen and is concerned with spermatogenesis and sperm maturation. PMID- 2986488 TI - The effect of verapamil and EGTA on the rat phrenic nerve-hemidiaphragm preparation. AB - Relatively high concentrations of verapamil or EGTA [ethylene glycol-bis (beta aminoethyl ether) N, N, N',N'-tetra acetic acid] inhibit contraction (P) of the rat phrenic nerve-hemidiaphragm preparation elicited by direct or indirect stimulation. The inhibitory effect of verapamil is greater (P less than 0.002) with direct (I50 = 26.3 +/- 1.7 microM) than indirect = I50 = 37.6 +/- 1.9 microM) stimulation. For EGTA the reverse is true: I50 is 1320 +/- 80 microM with direct and 1100 +/- 60 microM with indirect stimulation. The greater than 90% verapamil-induced depression of P can only be partially reversed by washout. Increasing the [Ca2+] or the addition of 4-aminopyridine (4AP) has insignificant antagonist effect. Except for the antagonism by 4AP during direct stimulation, the EGTA-induced depression of P is better antagonized by washout, increase of the [Ca2+], or the addition of 4AP than that caused by verapamil. Neostigmine did not antagonize the depression of P caused by either verapamil or EGTA. The findings presented indicate that the primary site of action of verapamil is postjunctional and that of EGTA is prejunctional. PMID- 2986489 TI - Therapeutic implications of modifying endogenous serotonergic analgesic systems. AB - Basic research strongly implicates the neurotransmitter serotonin as a modulator of endogenous analgesic systems. Recently, clinical strategies have been developed to activate endogenous serotonergic systems as a therapeutic approach to pain control. This paper reviews the biochemistry, anatomical distribution, and physiologic functions of serotonin. The evidence reviewed suggests that precursor loading to increase brain serotonin levels and administration of serotonin receptor inhibitors and serotonin receptor agonists may lead to novel methods of pain control and the development of useful analgesic drugs. PMID- 2986490 TI - Combined asthma and alveolitis due to diphenylmethane diisocyanate (MDI) with demonstration of no crossed respiratory reactivity to toluene diisocyanate (TDI). AB - Two workers, who developed asthmatic symptoms, were studied with inhalation provocation tests using diphenylmethane diisocyanate (MDI) and toluene diisocyanate (TDI). The subjects showed specific asthmatic reactions to MDI challenge (more than 20% fall in FEV1), and one also had an alveolar response. Alveolitis was suggested by fever, basal crackles, increased neutrophil counts in venous blood and in bronchoalveolar lavage. The asthmatic reaction to MDI challenge was associated with an increase in airway responsiveness to methacholine in both subjects. We conclude that MDI is a cause of asthma and/or hypersensitivity pneumonitis in workers exposed to diphenylmethane diisocyanate. PMID- 2986491 TI - Mastitis associated with ovine progressive pneumonia virus infection in sheep. AB - Eighteen ewes were experimentally infected with ovine progressive pneumonia virus and their mammary glands were examined for lesions and virus at 2.5 to 10 years postinoculation. Lesions were seen in 14 of 18 sheep; virus was isolated from 4 of 8 sheep. Lesion consisted of an interstitial accumulation of lymphocytes with periductal lymphoid nodules, and epithelial vacuolation and necrosis at the site of the lymphoid nodules. PMID- 2986492 TI - Production of superoxide anion by bovine pulmonary macrophages challenged with soluble and particulate stimuli. AB - The effects of opsonized zymosan, phorbal myristate acetate, and live Pasteurella haemolytica on superoxide anion production by bovine pulmonary macrophages were determined. The anion responses were dose-dependent for all stimuli, except for unopsonized P haemolytica. The effect of viable P haemolytica on macrophage viability was related to bacterial dosage and the presence of opsonizing antibody. Superoxide responses varied directly with the dose of opsonized live P haemolytica, but indirectly with macrophage viability. PMID- 2986493 TI - Morphogenesis of canary poxvirus and its entrance into inclusion bodies. AB - A virus isolated from a natural outbreak of canarypox was replicated on the chorioallantoic membranes of chicken embryos, and its ultrastructure and development were observed. Electron microscopy of thin sections of pocks produced on the chorioallantoic membranes revealed a variety of developmental forms which appear similar to those demonstrated in studies of vaccinia, ie, viroplasm or viral factories; immature, undifferentiated virions partially enclosed by membranes; completely enclosed nondifferentiated spherical or oval virions; immature virions with discrete nucleoids; and the more compact brick-shaped mature virions. Two types of A-type inclusions were noted: those with virions around the periphery, and those filled with virus particles. The appearance of mature viruses within the inclusion bodies and different stages of viruses outside the inclusion indicate that in a course of development, maturing poxvirus may enter the inclusion bodies as they acquire surface tubules on their envelopes. Mature virions also were seen budding out of the cell membrane, apparently enveloped in a portion of the membrane. Studies showing the entrance of poxvirus into inclusion bodies have not been reported. In this report, electron micrographs are shown of viruses entering inclusion bodies. PMID- 2986494 TI - Severe clinical disease induced in cattle persistently infected with noncytopathic bovine viral diarrhea virus by superinfection with cytopathic bovine viral diarrhea virus. AB - Eight healthy cattle that were persistently infected with noncytopathic bovine viral diarrhea virus (BVDV) were inoculated with cell culture fluids that contained noncytopathic or cytopathic BVDV. A severe disease occurred after inoculation with cytopathic BVDV. The clinical signs, lesions, and immune response were consistent with those of clinical BVDV infections. PMID- 2986495 TI - Dose-response evaluation of a genetically engineered foot-and-mouth disease virus polypeptide immunogen in cattle. AB - Four groups of 9 cattle each were vaccinated with 10, 50, 250, or 1,250 micrograms of foot-and-mouth disease (FMD) virus A12 VP1 fusion protein that was produced in Escherichia coli and emulsified in an oil adjuvant. The groups given the 10 and 50 micrograms of antigen were revaccinated at 15 weeks and were challenge exposed at 30 weeks; 5 of 9 and 7 of 9 cattle, respectively, were protected from FMD virus infection. The remaining 2 groups, vaccinated with 250 or 1,250 micrograms of antigen, were revaccinated at 32 weeks and were challenge exposed at 45 weeks; 8 of 9 and 9 of 9 cattle, respectively, were protected. The results indicated that the biosynthetic polypeptide FMD vaccine was effective using vaccination intervals frequently followed with conventional whole-virus vaccines. PMID- 2986496 TI - Enzymatic and acidic sensitivity profiles of selected virulent and attenuated transmissible gastroenteritis viruses of swine. AB - This study identifies the in vitro differences (markers) between virulent and attenuated transmissible gastroenteritis (TGE) viruses. Exposure of virulent Miller strain and attenuated Purdue strain TGE viruses to a spectrum of acidities indicated that the Miller strain was more stable at pH 2. Acidities at or above pH 3 did not reduce viral infectivity of either strain. When virulent and attenuated viruses were exposed to gastric fluids of either fed or fasted swine, there was a similar degree of sensitivity. Carboxypeptidase B, alpha-amylase, and alkaline phosphatase present in porcine small intestinal fluids did not cause a significant difference in sensitivity between virulent and attenuated virus isolates. The digestive enzymes: trypsin, alpha-chymotrypsin, pancreatin, peptidase, and carboxypeptidase A did not (or only slightly) inactivate virulent Miller strain TGE virus, but greatly reduced infectivity of attenuated viruses (Purdue strain and TGE vaccine virus isolates). The attenuated strains were significantly more sensitive to small intestinal fluids from both fasted and fed adult swine. Differential sensitivities between virulent and attenuated TGE viruses to digestive fluids from stomach and small intestine further substantiate the notion of differential susceptibility to small intestinal proteases as a correlate of viral virulence. PMID- 2986497 TI - Effect of virus-induced destruction of villous epithelium on intestinal secretion induced by heat-stable Escherichia coli enterotoxins and prostaglandin E1 in swine. AB - Villous atrophy and crypt hyperplasia were induced in the jejunal epithelium of thirteen 3-week-old pigs by inoculation with transmissible gastroenteritis virus. The responses (changes in net fluid movement) induced in ligated intestinal loops of these pigs by intraloop injections of prostaglandin E1 (PGE1) or Escherichia coli broth culture filtrates containing either or both E coli heat-stable enterotoxins (STa and STb) were compared with the responses induced by these preparations in littermates not inoculated with virus. Villous atrophy was associated with a marked decrease in response to preparations containing STa, STb, or STa + STb, but the response to PGE1 was undiminished. These results were consistent with the reports of others that the response to cyclic adenosine monophosphate-mediated secretogogues (PGE1) is a function of crypt epithelium; however, the present results also suggest that the secretory response to STa and to STb is dependent on the integrity of the villous epithelium. In the present study, loss of villous epithelium was associated with loss of response to STa and STb, but not to PGE1. PMID- 2986498 TI - Brachygnathia in a herd of Angus cattle. AB - Seven cases of brachygnathia in a single calf crop (28 calves) were observed in an Angus herd. Two of the affected calves, 1 female and 1 male, were necropsied and anatomically described. Breeding histories and results of pedigree analysis involving the 7 affected calves indicated the cause of the defect to be genetic with the mode of inheritance that of a single autosomal recessive gene. PMID- 2986499 TI - Application of the peroxidase-antiperoxidase procedure to the localization of pituitary hormones and calcitonin in various domestic animals and human beings. AB - Specific cell populations in the pituitary glands of the rat, cat, pig, and human being were positive for thyroid-stimulating hormone (TSH), luteinizing hormone (LH), and follicle-stimulating hormone (FSH). When reacted with prediluted rabbit anti-human TSH, LH, and FSH, antisera were not positive for the demonstration of these hormones in the horse, cow, or dog. Immunocytochemical staining was obtained in the horse, cow, and dog by the use of a primary antiserum against a specific beta-subunit of bovine TSH. The immunocytochemical staining of TSH, LH, FSH, adrenocorticotropic hormone, growth hormone, prolactin, and calcitonin was examined by the peroxidase-antiperoxidase method, using standard commercially available kits. All species examined had a strong positive reaction in specific pituitary cell populations for adrenocorticotropic hormone, growth hormone, and prolactin. Sections of normal thyroid gland tissue had positive staining of C cells containing calcitonin at the dilution of 1:100 of the primary antibody in the rat, horse, cow, dog, cat, pig, and human being. PMID- 2986500 TI - Reduction of neutrophil-mediated injury to pulmonary endothelial cells by dapsone. AB - The presence of activated neutrophils in the alveolar structures is thought to contribute to parenchymal cell injury in various acute and chronic lung disorders. This study indicates that dapsone (30 micrograms/ml), an agent with anti-inflammatory properties, can significantly reduce neutrophil-mediated injury to 51Cr-labeled bovine pulmonary artery endothelial (BPAE) cells with a reduction in the injury (expressed as a cytotoxic index) from 65 +/- 3 to 33 +/- 3 (p less than 0.001). Dapsone was unable to protect 51Cr-labeled BPAE cells injured by the chemical generation of superoxide, hydrogen peroxide, or neutrophil-derived, myeloperoxidase-dependent hypohalite ion. In contrast, dapsone significantly inhibited the respiratory burst of the neutrophil, with a reduction in the generation of superoxide, hydrogen peroxide, and conversion of nitroblue tetrazolium to formazan (p less than 0.01, all comparisons). Thus, dapsone appears to protect lung parenchymal cells such as endothelial cells from neutrophil-mediated injury by directly inhibiting the respiratory burst of the neutrophil, with a consequent diminution in the generation of toxic, oxygen derived radicals. PMID- 2986501 TI - Direct and indirect effects of leukotriene D4 on the pulmonary and systemic circulations. AB - We investigated direct (vascular leukotriene receptor stimulation) and indirect (generation of cyclooxygenase metabolites) hemodynamic effects of leukotriene D4 (LTD4) in 6 conscious sheep. Pulmonary artery, pulmonary arterial wedge and systemic arterial pressures, and cardiac output were measured. From these parameters, pulmonary vascular resistance (PVR) and systemic vascular resistance (SVR) were calculated before and immediately after a rapid injection of LTD4 into the pulmonary artery. Injection of 0.1 micrograms/kg of LTD4 increased mean PVR to 421% of baseline (p less than 0.001). It produced a biphasic effect on SVR that, after an initial decrease of 18% (p less than 0.05), increased to 143% of baseline (p less than 0.05). Both PVR and SVR returned to baseline within 10 min. The same results were obtained when the dose of LTD4 was increased to 0.5 micrograms/kg. Dose-response curves with increasing doses of LTD4 )0.025 micrograms/kg to 0.5 micrograms/kg) revealed that the optimal dosage for maximal effect was 0.1 micrograms/kg. The effects of LTD4 (0.1 micrograms/kg) on the pulmonary circulation were completely blocked by the SRS-A antagonist, FPL-57231, as well as by indomethacin. In the systemic circulation, FPL-57231 blocked the biphasic effects of LTD4 on SVR, whereas indomethacin prevented the initial decrease without attenuating the subsequent increase in mean SVR (135% of baseline, p less than 0.05). We conclude that there are direct and indirect hemodynamic effects of LTD4: the systemic vasoconstrictor response is directly related to vascular leukotriene receptor stimulation, whereas activation of cyclooxygenase pathway products is responsible for the pulmonary vasoconstrictor and systemic depressor responses. PMID- 2986502 TI - Production of arachidonic acid metabolites by macrophages exposed in vitro to asbestos, carbonyl iron particles, or calcium ionophore. AB - Consequent to asbestos deposition, alveolar macrophages (AM) accumulate at alveolar duct bifurcations where they phagocytize fibers. Because phagocytosis can stimulate the release of arachidonic acid (AA) metabolites, the possibility that secretion of these powerful mediators of inflammation might be induced by chrysotile asbestos was investigated in vitro. Rat AM were treated in vitro with chrysotile asbestos, and the cyclooxygenase products--prostaglandins, thromboxane B2 (TXB2), 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT)--and lipoxygenase products--leukotrienes (LT), hydroxyeicosatetraenoic acids (HETE)--secreted in the medium were isolated by high-performance liquid chromatography. Composition of the AA metabolites released was compared with that from those stimulated by the calcium ionophore A 23187 (20 microM) and by another particulate phagocytic stimulus, i.e., carbonyl iron beads. Calcium ionophore stimulation induced a marked release of various AA metabolites in the medium from both the cyclooxygenase pathway (HHT, TXB2, and PGE2, in decreasing quantities, respectively) and the lipoxygenase pathway (LTB4, 5-HETE, 12-HETE, and LTC4). The major product was LTB4. Treatment of the macrophages with asbestos fibers induced the release of a similar array of AA metabolites, although there were smaller amounts of LTC4 and 12-HETE, but increased quantities of PGF2 alpha. A time course study showed a steady increase in metabolite production for 1 h, followed by a plateau. In addition, the amount of metabolites released was dependent on asbestos concentrations. Phagocytosis of iron beads induced the secretion of the same metabolites as asbestos stimulation, but in larger quantities, probably reflecting the lack of cytotoxicity of the particle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986503 TI - Localization of ectopic parathyroid adenomas by the technetium-thallium subtraction scan. AB - Among numerous methods utilized for preoperative localization of hyperfunctioning ectopic parathyroid tissue, the technetium-thallium scan appears to be a major advance. Eight patients are presented where this method located ectopic parathyroid tissue in the neck, in the mediastinum, within the pericardium, and within the thyroid gland. To date, these findings have been confirmed operatively in five of the eight patients. On the basis of our limited experience, the authors believe that this is a valuable method in the preoperative localization of ectopic parathyroid tissue, especially in patients with persistent or recurrent hyperparathyroidism following a thorough neck exploration. PMID- 2986504 TI - Thrombocytopenic purpura in narcotics addicts. AB - Since November 1982 we have seen an association of thrombocytopenic purpura with chronic narcotic addiction in 70 patients with a mean platelet count of 53000 +/- 4000 (SE); 33 had stopped taking intravenous drugs for an average of 21.2 +/- 4.7 months; 13 of 15 had elevated antibody titers for a virus related to the acquired immunodeficiency syndrome. Platelet-bound IgG, IgM, and complement levels were 16.7-, 5.6-, and 3.1-fold greater than control values, respectively, and 2.6-, 1.9-, and 2.4-fold greater than values in 25 patients with classic autoimmune thrombocytopenic purpura studied at the same time. Thirty-three of thirty-six addicts had elevated circulating immune complexes, whereas 8 patients with autoimmune thrombocytopenia had no elevation. Eleven of eighteen addicts had positive serum platelet-reactive IgG titers, compared to 5 of 19 patients with classic autoimmune thrombocytopenia. The platelet-reactive IgG in sera of addicts was composed of 7S IgG antibody as well as high molecular weight (immune complex) IgG. Thus, chronic addicts appear to have a new immunologic platelet disorder associated with the presence of 7S IgG antiplatelet antibody, like patients with classic autoimmune thrombocytopenic purpura, and immune complex associated nonspecific platelet IgG, like male homosexual patients with thrombocytopenia. PMID- 2986505 TI - Bronchoalveolar lavage and transbronchial biopsy for the diagnosis of pulmonary infections in the acquired immunodeficiency syndrome. AB - The efficacy of bronchoalveolar lavage and transbronchial biopsy in diagnosing lung infection was determined in 276 fiberoptic bronchoscopic examinations done on 171 patients with known or suspected acquired immunodeficiency syndrome. Of 173 pathogens (Pneumocystis carinii, cytomegalovirus, Mycobacterium avium intracellulare, Cryptococcus neoformans, M. tuberculosis, Coccidioides immitis, and Histoplasma capsulatum) identified during the initial evaluation or in the subsequent month, the initial bronchoscopic examination detected 166 (96%). Bronchoalveolar lavage and transbronchial biopsy had sensitivities of 86% and 87%, respectively. When bronchoscopy included both bronchoalveolar lavage and transbronchial biopsy, the yield for all pathogens was 98% and the sensitivity for P. carinii infections was 100%. Follow-up for at least 3 weeks after examination failed to detect any additional false-negative results. Fiberoptic bronchoscopy is extremely accurate for the detection of pathogens in patients with the acquired immunodeficiency syndrome, especially when bronchoalveolar lavage and transbronchial biopsy are combined. In patients at high risk of complications from transbronchial biopsy, bronchoalveolar lavage is sufficiently accurate to be used alone. PMID- 2986506 TI - Acquisition of antibody to lymphadenopathy-associated virus in patients with classic hemophilia (factor VIII deficiency). AB - Antibody to lymphadenopathy-associated virus (LAV) was assayed in 461 serum and plasma samples that had been obtained from 149 patients with classic hemophilia and 64 controls and stored for periods as long as 18 years. No control or patient samples obtained before 1980 contained antibody to this retrovirus. The prevalence of antibody to LAV in the patient samples rose from 15% (2 of 13) in 1980 to 62% (18 of 29) in 1984. During this time, none of 8 untreated hemophiliacs and none of 26 hemophiliacs treated solely with cryoprecipitates had antibody to LAV. In contrast, the prevalence of antibody to LAV among hemophiliacs treated with lyophilized antihemophilic factor rose from 25% (2 of 8) in 1980 to 78% (18 of 23) in 1984. These seropositive hemophiliacs had fewer OKT4 helper cells and lower proliferative responses to mitogen than similarly treated seronegative patients. Treatment with locally prepared cryoprecipitates was not associated with serologic evidence of virus exposure. PMID- 2986507 TI - Theophylline toxicity and the beta-adrenergic system. AB - After ingestion of 12 g of theophylline caused severe toxicity in a young woman, we developed an experimental canine model to study human theophylline toxicity. Our study involved four anesthetized dogs given theophylline in a continuous intravenous drip for 180 minutes in one of four protocols. The protocols included a low-dose infusion (400 mg/h), a high-dose infusion (1000 mg/h), a high-dose infusion with beta-blockade induced by propranolol at 125 minutes after infusion, and a high-dose infusion while maintaining beta-blockade with propranolol throughout the experiment. Toxic levels of theophylline were associated with hypokalemia, hypophosphatemia, hyperglycemia, metabolic acidosis, and hypotension in both the patient and the experimental series. These effects were either prevented or partially reversed after induction of beta-blockade with propranolol. Very high levels of theophylline were associated with elevated levels of norepinephrine and epinephrine in the animals. PMID- 2986508 TI - Oral acyclovir therapy for mucocutaneous herpes simplex virus infections in immunocompromised marrow transplant recipients. AB - In a randomized, double-blinded, placebo-controlled trial, we compared the therapeutic effect of oral acyclovir (400 mg five times daily for 10 days) with that of placebo in patients with marrow transplants and culture-proven recurrent mucocutaneous herpes simplex. Twelve patients received acyclovir and nine received placebo. Acyclovir treatment significantly shortened the median duration of viral shedding, new lesion formation, and time to first decrease in pain, resolution of pain, 50% healing, and total healing. These results compared favorably with those previously obtained with intravenous or topical acyclovir preparations. Oral acyclovir is highly effective for the treatment of recurrent mucocutaneous infections caused by herpes simplex virus in immunocompromised patients. PMID- 2986509 TI - NIH conference. The acquired immunodeficiency syndrome: an update. AB - The acquired immunodeficiency syndrome continues to be a major public health problem in the United States, and recently its spread worldwide has accelerated. The syndrome is caused by a human retrovirus transmitted by sexual contact and via blood or blood products. The virus has been isolated, characterized, and cloned, and in addition to its presence in blood, it has been found in body tissues and fluids including brain, semen, and saliva. Although the syndrome in the United States is still largely confined to male homosexuals and intravenous drug users, there is increasing evidence, particularly from Zaire, that the virus can be spread by heterosexual contact. Attempts at immune reconstitution with lymphocytes and lymphokines have resulted in some transient improvement in immune function but without clinical effect, indicating the need for specific anti retroviral therapy in combination with immune reconstitution. PMID- 2986511 TI - Rheumatoid arthritis and Epstein-Barr virus. PMID- 2986510 TI - Immunoglobulins in the acquired immunodeficiency syndrome. PMID- 2986512 TI - Epstein-Barr virus and unexplained illness. PMID- 2986514 TI - TORCH serologies and specific IgM antibody determination in acquired and congenital infections. AB - Except for rubella testing, routine TORCH serology screens in prenatal care are of little use. Individual TORCH tests may, however, be useful based on the clinical presentation and history of the patient. The laboratory test of choice for diagnosing cytomegalovirus (CMV) and herpes simplex virus (HSV) infections is culture isolation for the virus. The presence for specific IgM antibodies in neonates is diagnostic of congenital infection. In adults, IgM antibody results should be interpreted along with the clinical findings and history of the patient. IgM antibodies may persist for months and even years and may be detected during reactivation of latent virus infections. Serum fractionation should always be performed in IgM antibody testing to avoid false positive results owing to rheumatoid factors and false negative results owing to competing levels of specific IgG antibodies. With a single serum specimen, specific IgM antibody detection may be helpful in differentiating between a recent versus past infection. PMID- 2986513 TI - Adverse drug reactions in the newborn. AB - Adverse drug reactions have been noted to occur in one of three newborns admitted to intensive care units. The factors associated with adverse drug reactions and the adverse reactions to commonly used medications in neonatal intensive care units are discussed. Methods of preventing or reducing these undesired effects are suggested. PMID- 2986515 TI - Idiotypy of catecholamine-binding proteins. AB - The idiotypic and antiidiotypic response to alprenolol, a beta-adrenergic antagonist, was studied both in rabbits and in mice. Rabbit polyclonal anti alprenolol antibodies showed binding properties for catecholamine analogs, agonists as well as antagonists, similar to those of the beta-adrenergic receptors. The variability of the anti-alprenolol response was studied by using mouse monoclonal antibodies specific for alprenolol. While the binding properties showed great variations in affinity, the response seemed restricted to the heavy chain classes gamma 1 and gamma 2a. N-terminal sequencing of the light and heavy chains and restriction maps of the corresponding genes suggest that the antibodies use particular subgroups infrequently found in antibodies specific for other antigens. The cyclical antiidiotypic response in rabbits immunized with polyclonal antibodies and in mice immunized with monoclonal antibodies were compared. The response of the latter was dependent on the choice of the monoclonal antibody used to elicit the antiidiotypic response. Finally, the agonist-like properties of a monoclonal antiidiotypic antibody directed against one of the monoclonal anti-alprenolol antibodies were studied extensively. The ability to recognize beta-adrenergic receptors was documented by Western blot and direct immunoprecipitation and visualized by immunofluorescence. The antiidiotypic antibody stimulated catecholamine-sensitive adenylate cyclase and this effect was blocked by the beta-adrenergic antagonist propranolol. PMID- 2986516 TI - Evolution within the multigene family coding for the class I histocompatibility antigens: the case of the mouse t-haplotypes. AB - The t-haplotypes of the house mouse are defined by several genetic events which involve about one-third of chromosome 17 and include the major histocompatibility complex (MHC). One of the primary features of complete t-haplotypes is the nearly complete suppression of recombination between a t-chromosome and a wild-type chromosome 17. If we assume that all genetic exchanges are abolished because of the inhibition of recombination, the class I genes of the MHC can be used as vicinity markers of t-haplotypes. We have studied the H-2K, D and L genes of 10 independant t-haplotypes and have found that their H-2K genes are rather polymorphic, although always very similar or identical to H-2K genes of laboratory mice. By contrast, the DL-Tla region is nearly constant. It is suggested that the t-haplotypes reveal the existence of several mechanisms involved in the evolution of the MHC, one of which is possibly the greater "mobility" of H-2K genes in the genome. PMID- 2986517 TI - Allelism in the HLA class I multigene family. AB - Recent data on the structure of functional HLA class I genes shows that, at least at the HLA-A locus, the allelic genes are more related to each other than to HLA genes from other loci. This "A-ness" is discernible at the protein sequence level but much more evident when nucleotide sequences are compared; the homology is particularly striking in the 3' non-coding region. Genes coding for the same HLA specificity in different genetic backgrounds show no obvious difference, although in one case the 3' flanking regions are clearly different; the restriction maps around the HLA-A3 and HLA-AW24 genes are also compared to see whether the chromosomal environments of these two genes are recognizably similar. PMID- 2986518 TI - Induction of differentiation in neoplastic cells. AB - There is now clear evidence that cells cultured from human and animal tumours can be induced to differentiate in vitro by recognised hormones, regulatory peptides, polar solvents and cytotoxic drugs. Examples can be found from several different types of tumour with the bulk of the data deriving from neuroblastoma and myeloid leukaemia. There is no clear correlation of inducer with cell type, other than some specific peptides like MSH, and agents such as dimethyl sulphoxide and dexamethasone have wide ranging activity. Steroid hormone action may require interaction between different cell types, and the inability of tumours to differentiate in situ may implicate reduced cell-cell interaction, possibly due to degradation of extracellular matrix, or to alteration of the stromal phenotype by tumour-derived factors such as peptides or prostaglandins. When differentiation has been demonstrated, it has been possible, in some cases, to correlate increased differentiation with reduced malignancy by in vitro characterisation or tumorigenicity. Conditions which induce differentiation in rat mammary carcinoma and mouse myeloma also reduce tumour growth in vivo. Clinical trials have not provided any conclusive evidence for a therapeutic benefit so far, but relatively few trials have been carried out. There is clearly a need for further investigation both in vitro and in vivo to select optimal conditions and combinations of agents for clinical evaluation. PMID- 2986519 TI - The biology of cell death in tumours. AB - The principal objective of this article is to present evidence for the numerical and biological importance of cell death in tumour growth and regression. Much of this death has the morphology of apoptosis, a process observed elsewhere in biology in a variety of circumstances. Recent studies on the mechanism of apoptosis in non-neoplastic cells suggest that a relatively small number of specific intracellular changes are involved. Two of these are described: endonuclease activation within the nucleus, which coincides with the characteristic morphological changes there, and altered expression of surface carbohydrates, which may be responsible for the swift recognition and phagocytosis of apoptotic cells. It seems probable that similar events occur during the apoptosis of tumour cells. Further knowledge of the control of these endogenous mechanisms for effecting cell death could lead to powerful new means of controlling the growth of tumours. PMID- 2986520 TI - The biology of papillomaviruses and their role in oncogenesis. AB - A large number of different papillomavirus types have been identified. Several viruses often infect the same species and each virus is associated with a defined tissue. Recent evidence has shown that certain benign lesions can undergo malignant transformation in both animals and humans in response to genetic or environmental factors. Fine mapping of the structure and function of the viral genome may enhance our understanding of the interaction between the virus and the cofactors involved in malignant transformation. PMID- 2986521 TI - Feline leukaemia viruses: molecular biology and pathogenesis. AB - The feline leukaemia virus (FeLV) group represents one of the most important viral pathogens of the domestic cat. In addition, this virus - host system is one of the major experimental models for retroviral pathogenesis. Under natural conditions, the virus is horizontally transmitted through the cat population. The outcome of infection depends on a variety of factors including the virus does encountered and the age and immune status of the host. FeLVs can establish persistent infection, either overt or latent. Degenerative diseases of the haemopoietic system are the most common result of persistent infection and immunosuppression with secondary infection accounts for more deaths than does neoplastic disease. However, more is known about the molecular mechanisms of oncogenesis in this system and there are now numerous examples of field case tumours where FeLV has transduced an oncogene or acted as an insertional mutagen. The factors affecting the relative frequency of these mechanisms are considered as is the possibility that recombinant env gene recombinants play a role in FeLV pathogenesis. PMID- 2986522 TI - Two oncogenes in avian carcinoma virus MH2: myc and mht. AB - The 5.2-kilobase (kb) RNA genome of avian carcinoma virus MH2 has the genetic structure 5' - delta gag (0.2 kb)-mht (1.2 kb)-myc (1.4 kb)-c(0.4 kb)-poly (A) (0.2 kb)-3'. delta gag is a partial retroviral core protein, mht and myc are cell derived MH2-specific sequences, and c is the 3'-terminal retroviral vector sequence. the following results were obtained from the complete nucleotide sequences of the mht and myc genes in MH2. (i) delta gag-mht forms a hybrid gene with a contiguous reading frame of 2682 nucleotides that terminates with a stop codon near the 3' end of the mht gene. The 3' 969 nucleotides of mht up to the stop codon are 80% sequence related to the onc-specific raf sequence of murine sarcoma virus 3611 (MSV 3611) (94% homologous at the deduced amino acid level). (ii) The myc coding region in MH2 is preceded by 181 nucleotides derived from the intron immediately upstream from the second exon of the chicken cellular proto myc gene, followed by an RNA splice acceptor site shared with the proto-myc gene, followed by an RNA splice acceptor site shared with the proto-myc, beyond which it is colinear up to a 3'-termination codon and 40 noncoding nucleotides with the myc sequences of avian retrovirus MC29 and chicken proto-myc. Thus, myc forms, together with a 5' retroviral exon, a second MH2-specific gene. It is concluded that MH2 contains two genes with oncogenic potential, the delta gag-mht gene, which is closely related to the delta gag-raf transforming gene of MSV 3611, and the myc gene, which is related to the transforming gene of MC29. Furthermore, it may be concluded that the cellular proto-onc genes, which on sequence transduction become viral onc genes, are a small group because among the 19 known onc sequences, 5 are shared by different taxonomic groups of viruses of which the mht/raf homology is the closest so far. PMID- 2986523 TI - An ultrastructural morphometric analysis of normal human mammary tissue and human breast cancer. AB - An ultrastructural morphometric study of normal and tumourous adult human mammary tissue is presented. The data show a characteristic numerical pattern for the different neoplastic stages of breast tissues. By applying a special data comparison program a standardized diagnostic and prognostic system appears possible through the morphometric analysis of ultrathin epon sections of surgically obtained material. PMID- 2986524 TI - Functional characteristics of transplantable rat pituitary tumor MtT SA5 inducing pronounced adrenal hyperplasia. AB - Transplantable rat pituitary tumor MtT SA5 is characterized by the induction of pronounced adrenal enlargement. In spite of remarkable elevation of plasma ACTH levels, serum corticosterone levels in hypophysectomized tumor-bearing rats were lower than those in intact control rats. No immunohistochemical alterations were seen in corticotrophs of the pituitary in tumor-bearing rats. Ratio of bioactive ACTH (Bio-ACTH) to immunoreactive ACTH (Ir-ACTH) in the tumor tissues was 0.2%, being extraordinarily lower than in the pituitary tissues (23%). Gel chromatography of the tumor tissue showed large molecular weight forms of Ir ACTH, which seem to result from abnormal processing of proopiomelanocortin, and are responsible for the lower levels of Bio-ACTH compared to Ir-ACTH. The MtT SA5 tumor is suggested to secrete ACTH-related peptides which induce pronounced adrenal enlargement with little or no stimulation of glucocorticoid production. PMID- 2986525 TI - Inflammatory myopathies: Part 1. AB - The inflammatory myopathies have diverse clinical and pathological features and multiple etiologies. Some are confined to a single muscle or group of muscles (e.g., orbital myositis and localized nodular myositis) while others are diffuse. Infective forms may be due to viral, bacterial, fungal, protozoal, or parasitic organisms. Viruses may cause acute self-limited forms of myositis and have been isolated from muscle in some cases of acute rhabdomyolysis and inclusion body myositis. They have also been implicated in some cases of congenital myopathy and in polymyositis and dermatomyositis, but there is no evidence of viral invasion of muscle in these conditions. In polymyositis and dermatomyositis there are derangements in humoral and cellular immune function, and recent evidence suggests an underlying disturbance of immunoregulation. The roles of genetic factors, drugs, and Toxoplasma infection have been under scrutiny. There is increasing recognition of immunological and pathological differences in polymyositis and juvenile and adult dermatomyositis, and in cases with associated connective tissue diseases, suggesting different underlying pathogenetic mechanisms. Inclusion body myositis, eosinophilic myositis, and granulomatous myositis can be separated from the other idiopathic inflammatory myopathies because of distinctive clinical and pathological features and this may also reflect different mechanisms of muscle injury. Recent developments in the treatment of the idiopathic inflammatory myopathies include the use of plasmapheresis and total-body irradiation in cases that are resistant to corticosteroids and immunosuppressive drugs. PMID- 2986526 TI - Axonal attenuation and secondary segmental demyelination in myeloma neuropathies. AB - The sural nerves of 5 patients with osteosclerotic myeloma and polyneuropathy, of 3 patients with multiple myeloma and polyneuropathy, and of 6 healthy subjects were studied by neuropathological, morphometric, and teased-fiber approaches to assess cellular (Schwann cell or axon) vulnerability and to explore the mechanism of segmental demyelination. As compared with controls, the nerves of both types of myeloma neuropathy demonstrated a statistically significant and marked loss of myelinated fibers and increased frequencies of axonal degeneration among teased fibers at statistically significant levels. The peaks of diameter histograms of myelinated fibers of osteosclerotic myeloma/polyneuropathy nerves were displaced to smaller diameter categories, suggesting fiber atrophy. Segmental demyelination and remyelination was clustered, as found in secondary demyelination. Large- and intermediate-diameter myelinated fibers of osteosclerotic myeloma/polyneuropathy nerves had significantly smaller axon calibers relative to myelin spiral length seen in electron micrographs. The loss of myelinated fiber axons, the shift of the peaks of diameter histograms to smaller sizes, the lack of noticeable increased numbers of demyelinated axons, the clustered distribution of segmental demyelination, and the axonal attenuation suggest a special axonal or neuronal vulnerability and appear to provide an explanation for the observed segmental demyelination. Whether axonal attenuation has a perikaryeal or proximal axonal genesis now needs to be determined. PMID- 2986527 TI - Surgical management of tumors that produce gastrointestinal hormones. AB - Gastrointestinal hormones are produced by specialized endocrine cells found both in the pancreas and distributed throughout the gut. Tumors, both benign and malignant, produce excessive amounts of hormone and thereby cause characteristic syndromes. Surgical management must be primarily directed at tumor removal in order to effect cure of the syndrome, as well as to prevent complications of persistent tumor growth and, subsequently, metastasis. PMID- 2986528 TI - The molecular mechanism of insulin action. AB - Insulin initiates its action by binding to a glycoprotein receptor on the surface of the cell. This receptor consists of an alpha-subunit, which binds the hormone, and a beta-subunit, which is an insulin-stimulated, tyrosine-specific protein kinase. Activation of this kinase is believed to generate a signal that eventually results in insulin's action on glucose, lipid, and protein metabolism. The growth-promoting effects of insulin appear to occur through activation of receptors for the family of related insulin-like growth factors. Both genetic and acquired abnormalities in the number of insulin receptors, the activity of the receptor kinase, and the various post-receptor steps in insulin action occur in disease states leading to tissue resistance to insulin action. PMID- 2986529 TI - Hormonal regulation of glomerular filtration. AB - Technological advances within the last decade now permit examination of the effects of various vasoactive substances on glomerular hemodynamics and the filtration process. By modulating the vasomotor tone of the preglomerular and postglomerular arterioles, these substances influence the rate of plasma entering the glomerulus and the pressure within the glomerular capillaries. In addition, hormones appear to influence the permeability of glomerular capillaries by altering the surface area available for filtration. PMID- 2986530 TI - Acquired immunodeficiency syndrome: epidemiology, virology, and immunology. AB - Acquired immunodeficiency syndrome indicates opportunistic infection or unusual malignancy associated with a marked deficiency of cell mediated immunity in the absence of any other cause of the immune defect. This review examines epidemiology, virology, and immunology associated with this disorder. The syndrome appears to be caused by infection of at-risk individuals by a retrovirus that has tropism for a specific population of T lymphocytes. PMID- 2986531 TI - Chemistry of gap junctions. PMID- 2986532 TI - Oscillatory neural networks. AB - Despite the fact that a large number of neuronal oscillators have been described, there are only a few good examples that illustrate how they operate at the cellular level. For most, there is some isolated information about different aspects of the oscillator network, but too little to explain the whole mechanism. Two quite remarkable features do seem to be emerging from ongoing studies, however. One is that there are very few generalizable features common to neural oscillators. Many utilize reciprocal inhibitory circuits and endogenous burst generating currents to some extent. All that have been well worked out utilize a combination of both cellular and network properties, but little else in the way of common mechanism is noteworthy. Perhaps the most interesting aspect of recent work is the ability of a particular oscillator to produce a large repertoire of different outputs. This is separate and in addition to changes occurring via phasic sensory feedback. It is in fact a radical functional "rewiring" of the network in response to neuromodulators. The CPG circuits represent only the most basic form of a given pattern. Finally, concerning the role of sensory feedback in generating oscillatory patterns, the concept of the CPG as a group of neurons able to produce oscillatory patterns without any sensory feedback is, in our opinion, still valid. There is no doubt that some oscillators may be quite weak when isolated, but they can still produce bursts with firing sequences similar to those seen in vivo. The fact that sensory feedback can both control and enhance the oscillations has never been in doubt. Similarly, entrainment of the pattern by sensory feedback does not mean that the receptor is part of the generator, only that it has access to it (as do command and coordinating fibers). The real question remains: Can a group of cells produce an oscillatory pattern without phasic sensory input? We must answer this affirmatively even for the insect flight motor CPG, while emphasizing the fact that for this system sensory feedback plays a larger role than in most other CPGs. Most neural oscillators will probably fall on some continuum between those like insect flight, which need and use a large amount of phasic feedback, and those that can oscillate in a near normal manner without it. PMID- 2986533 TI - Antibody probes in the study of gap junctional communication. PMID- 2986534 TI - The nature and regulation of the insulin receptor: structure and function. AB - Native, cell-surface insulin receptor consists of two glycoprotein subunit types with apparent masses of about 125,000 daltons (alpha subunit) and 90,000 daltons (beta subunit). The alpha and beta subunits are derived from a single polypeptide precursor by one or more proteolytic cleavages. The predominant subunit configuration in the native insulin receptor is a disulfide-linked heterotetrameric structure containing two alpha and two beta subunits. The alpha and beta insulin-receptor subunits seem to have distinct functions such that alpha appears to bind hormone whereas beta appears to possess intrinsic tyrosine kinase activity. In detergent extracts, insulin activates receptor autophosphorylation of tyrosine residues on its beta subunit, whereas in the presence of reductant, the alpha subunit is also phosphorylated. Other physiologically relevant substrates of the insulin receptor tyrosine kinase in target cells, if any, have not yet been identified. In intact cells, insulin activates serine/threonine phosphorylation of insulin receptor beta subunit as well as tyrosine phosphorylation. The biological role of the receptor-associated tyrosine kinase is not known. Tyrosine phosphorylation, catalyzed by either autophosphorylation or purified src kinase, of insulin receptor beta subunit in vitro activates the receptor kinase activity, whereas dephosphorylation with alkaline phosphatase deactivates the receptor kinase. The insulin receptor kinase is regulated by beta-adrenergic agonists and other agents that elevate cAMP in adipocytes, presumably via the cAMP-dependent protein kinase. Such agents decrease receptor affinity for insulin and partially uncouple receptor tyrosine kinase activity from activation by insulin. These effects appear to contribute to the biological antagonism between insulin and beta-agonists. The insulin receptor kinase is also inhibited in intact cells by phorbol esters that mediate serine/threonine phosphorylation of the insulin receptor, presumably via the Ca++ phospholipid-dependent protein kinase. These data suggest the hypothesis that a complex network of tyrosine and serine/threonine phosphorylations on the insulin receptor modulate its binding and kinase activities in an antagonistic manner. PMID- 2986535 TI - Uptake of insulin and other ligands into receptor-rich endocytic components of target cells: the endosomal apparatus. PMID- 2986536 TI - Intracellular mediators of insulin action. PMID- 2986537 TI - The nature and regulation of the receptors for insulin-like growth factors. AB - Two subtypes of IGF receptors have been identified. Type I IGF receptors have a Mr greater than 300,000 and are composed of disulfide-linked 130,000-dalton (alpha) and approximately 90,000-dalton (beta) subunits. The alpha subunit binds hormone; the beta subunit appears to have intrinsic tyrosine kinase activity and to be autophosphorylated. Type I receptors preferentially bind IGF-I but also bind IGF-II and, more weakly, insulin. Type II IGF receptors consist of a 250,000 dalton protein that contains internal disulfide bonds but is not linked to other membrane components. Type II receptors bind IGF-II with higher affinity than IGF I. They do not interact with even very high concentrations of insulin. Type I IGF receptors and insulin receptors are homologous structures. They have similar subunit structure. Both receptors bind IGFs and insulin. They have similar (but not identical) antigenic determinants. Both receptors are downregulated by IGFs and insulin. Both receptors are affected in certain patients with genetically determined insulin resistance. Type II IGF receptors do not appear to be homologous to type I receptors. They differ in structure, peptide binding specificity, and antigenic determinants. Type II receptors do not appear to be downregulated. Although type II receptors appear to be phosphorylated in intact cells, they do not possess intrinsic tyrosine protein-kinase activity. Insulin acutely upregulates type II IGF receptors in intact rat adipose cells by effecting a redistribution of receptors cycling between a large intracellular pool and the plasma membrane. Insulin and the IGFs elicit the same biological responses, either by cross-reacting with one of the receptors for the heterologous ligand or by concurrent activation of convergent effector pathways by binding to the homologous receptor. Which mechanism is utilized appears to depend more on the tissue than on the biological response. Insulin desensitizes rat hepatoma cells to the actions of insulin and IGFs, mediated by both insulin and IGF receptors, by mechanisms distal to hormone binding and possibly common to IGF and insulin effector pathways. PMID- 2986538 TI - Actions of insulin-like growth factors. PMID- 2986539 TI - The nature and regulation of the receptors for pituitary growth hormone. AB - Recent developments have contributed greatly to our understanding of GH receptor structure and function; however, much remains to be done. One of the first priorities should be to elucidate the early events that follow activation of the GH receptor. The in vitro systems outlined in this review show promise in this regard; however, all three systems require further characterization. Future studies of the GH receptor in these and other systems will undoubtedly be facilitated by the monoclonal antibody generated against the GH receptor and by recently described cross-linking methodologies. Despite these recent advances, however, it is likely that detailed characterization of receptor structure and function awaits cloning of the GH-receptor gene(s). PMID- 2986541 TI - Nucleotide metabolism by endothelium. PMID- 2986540 TI - Myocardial nucleotide transport. AB - A thorough consideration of the evidence for striated muscle cell transmembrane nucleotide movement provides only equivocal support for adenine nucleotide specific translocation across cell membranes. It is obvious that nucleotide derived adenosine is taken up into cells in preference to free adenosine, and it is important to understand why this is so. The potential importance of released nucleotides to cell regulation justify studies to determine the source and release mechanism. PMID- 2986542 TI - Purine salvage pathways in myocardium. PMID- 2986543 TI - [Natural interferon inducers: double-stranded RNA of killer plasmids of Saccharomyces cerevisiae]. AB - Methods for isolation and purification of yeast double spiral RNA (dsRNA) are described. The most simple method includes reprecipitation of dsRNA in solutions of lithium chloride and its interphase distribution in the system of chloroform: isoamilic alcohol: water. Preparations with the content (w/w) of dsRNA from 30 to 90 per cent were obtained. They possess a high interferon-inducing activity and an antiviral effect on the experimental infection caused by the virus of mouse encephalomyocarditis. PMID- 2986544 TI - In vitro activities of bismuth salts against rotaviruses and other enteric viruses. AB - Bismuth salts inhibited plaque formation of all enteric viruses tested, which included four strains of rotavirus and one strain each of echovirus, reovirus, and poliovirus. The compounds had no direct virucidal effect at concentrations ranging from 0.025 to 2.5 mg/ml, and the inhibitory effects were observed only at concentrations slightly below those that caused extensive cytotoxicity. Plaque inhibition appeared to result from interference with host cell functions, with secondary inhibition of virus production. PMID- 2986546 TI - Enzymatic and nonenzymatic superoxide-generating reactions of isoniazid. AB - During the course of horseradish peroxidase-mediated oxidation of either o dianisidine or 2-2'-azino-di(3-ethyl-benzthiazoline-6-sulfonic acid) (ABTS), no O2 consumption took place. When isonicotinic acid hydrazide (isoniazid) (INH) was included in the reaction mixture, O2 was consumed in amounts linearly related to the INH concentration. Nicotinic acid hydrazide at equimolar concentrations induced lower rates of O2 consumption. Superoxide dismutase activated O2 consumption. At equimolar concentrations, INH, nicotinic acid hydrazide, and phenylhydrazine induced bleaching of p-nitrosodimethylaniline in the horseradish peroxidase mediation of ABTS oxidation. Bleaching was not inhibited by hydroxyl radical (. OH) scavengers. After a short lag period, INH reacted with NADH at alkaline pH to produce superoxide radical (O2-), as detected by superoxide dismutase-inhibitable Nitro Blue Tetrazolium reduction. Nicotinic acid hydrazide with NADH caused a lower rate of O2- production after a longer lag period than INH. PMID- 2986545 TI - Evidence for the generation of active oxygen by isoniazid treatment of extracts of Mycobacterium tuberculosis H37Ra. AB - Crude extracts of Mycobacterium tuberculosis H37Ra, an isonicotinic acid hydrazide (isoniazid) (INH)-susceptible strain which has peroxidase activity, catalyzed the production of catechol from phenol in the presence of INH and H2O2 as shown by the development of the 444-nm absorption peak of oxidized catechol product. Extracts of the INH-resistant strain of M. tuberculosis H37Ra, which has no peroxidase, did not catalyze the reaction. The rate of development of the 444 nm peak increased proportionately with increased superoxide dismutase concentrations. The hydroxyl radical (. OH) scavengers dimethylsulfoxide and mannitol inhibited the reaction. Isonicotinamide, isonicotinic acid, and nicotinic acid could not replace INH. PMID- 2986548 TI - Interfering with taste aversion learning in rats: the role of associative interference. AB - Six experiments with rats investigated the conditions under which one flavor interferes with aversion conditioning to a second, familiar flavor. Conditioning to the familiar flavor was weakest when the interference flavor was contiguous to lithium-induced toxicosis, novel, more intense, and strongly associated with toxicosis. In addition, conditioning to the familiar flavor was weakened even if multiple conditioning trials were used. The repeated finding of an inverse relationship between strength of aversion to the target and interference flavors is interpreted as support for an associative competition hypothesis of the interference effect. The possible relevance of the interference effect to the attenuation of taste aversions in cancer patients is discussed. PMID- 2986547 TI - Interaction of Epstein-Barr virus DNA polymerase and 5'-triphosphates of several antiviral nucleoside analogs. AB - The 5'-triphosphates of 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5 methyluracil, 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine, 1 (2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-methylcytosine, 9-[(2 hydroxyethoxy)methyl]guanine, and 9-(1,3-dihydroxy-2-propoxymethyl)guanine had lower Ki values for Epstein-Barr virus DNA polymerase than has been reported elsewhere for host DNA polymerase. Inhibition of DNA elongation by these analogs ranged from moderate to strong, suggesting that preferential incorporation of these analogs into DNA by virus DNA polymerase may contribute to antiviral selectivity. PMID- 2986549 TI - Identification of the components of a putative cytochrome bc1 complex in Rhodopseudomonas viridis. AB - Chromatophore membranes isolated from the bacteriochlorophyll b-containing, photosynthetic purple nonsulfur bacterium, Rhodopseudomonas viridis, have been shown to contain a Rieske iron-sulfur protein, a cytochrome similar to cytochrome c1, and also at least one b-type cytochrome. These observations suggest the presence of a previously undetected cytochrome bc1 complex in this bacterium. PMID- 2986550 TI - Regulation of steroidogenesis in rat Leydig cells in culture: effect of human chorionic gonadotropin and dibutyryl cyclic AMP on the synthesis of cholesterol side chain cleavage cytochrome P-450 and adrenodoxin. AB - Rat Leydig cells in primary culture were used as a model system to investigate the effects of human chorionic gonadotropin (hCG) and dibutyryl cyclic AMP (Bt2cAMP) on the synthesis of cholesterol side chain cleavage cytochrome P-450 (cytochrome P-450scc) and the iron-sulfur protein, adrenodoxin. Leydig cells isolated from the testes of mature rats were placed in monolayer culture in the absence of stimulatory factors for 8 days. HCG (10 mIU/ml) or Bt2cAMP (1 mM) were then added to some of the cultures and the incubations were continued for up to 48 h. Testosterone production was increased markedly in cells incubated with hCG or Bt2cAMP. A significant accumulation of pregnenolone in the medium of cells treated with Bt2cAMP was also observed. Both hCG and Bt2cAMP increased the rates of synthesis of cytochrome P-450scc and adrenodoxin. In hCG-treated cells the apparent rate of synthesis of cytochrome P-450scc was increased 13-fold over that of controls after 48 h of incubation; the rate of adrenodoxin synthesis was increased 4-fold by hCG treatment. In Bt2cAMP-treated cells the rate of synthesis of cytochrome P-450scc was 37-fold greater than that of control cells after 48 h of incubation; adrenodoxin synthesis was increased 36-fold over controls. In hCG- and Bt2cAMP-treated cells, the concentration of immunoreactive cytochrome P 450scc and adrenodoxin increased with increasing time of incubation, and were correlated with the stimulatory effects of these agents on cytochrome P-450scc activity and on total steroid production. The results of this study are indicative that the maintenance by LH/hCG of elevated levels of testosterone synthesis by the Leydig cell is mediated, in part, by induction of the synthesis of cytochrome P-450scc and its associated protein, adrenodoxin. Since Bt2cAMP had effects similar to those observed with hCG, it is suggested that the stimulatory effects of hCG on the synthesis of cytochrome P-450scc and adrenodoxin are mediated by increased cyclic AMP formation. PMID- 2986551 TI - Dual pathways of receptor-mediated cyclic GMP generation in NG108-15 cells as differentiated by susceptibility to islet-activating protein, pertussis toxin. AB - The cellular cGMP content increased in response to a variety of receptor agonists, which activate [e.g., prostaglandin (PG) E1, E2, and F2 alpha] or inhibit (e.g., alpha-adrenergic, muscarinic, and opiate agonists) adenylate cyclase in neuroblastoma X glioma hybrid NG108-15 cells. The responses were additive when PGF2 alpha and enkephalin were mixed. The inhibitory guanine nucleotide regulatory protein (Ni) is involved in adenylate cyclase inhibition; this function of Ni is lost when it is ADP-ribosylated by islet-activating protein (IAP), pertussis toxin [H. Kurose, T. Katada, T. Amano, and M. Ui (1983) J. Biol. Chem. 258, 4870-4875]. The cGMP rise induced by stimulation of the receptors linked to adenylate cyclase inhibition was also diminished by IAP; the time course and dose response for the IAP-induced diminution were the same between adenylate cyclase inhibition and cGMP generation. Ni thus appears to mediate guanylate cyclase activation as well as adenylate cyclase inhibition initiated via the same receptors. Melittin also increased cGMP. No additivity was shown when enkephalin and melittin were combined, suggesting that phospholipase A2 might play a role in Ni-mediated guanylate cyclase activation. On the other hand, the PGF2 alpha-induced cGMP rise was associated with increased incorporation of 32Pi into phosphatidylinositol; was not affected by cholera toxin, IAP or forskolin; and showed no additivity when combined with A23187, which increased cGMP by itself. PGs would occupy receptors linked to phosphatidylinositol breakdown, thereby increasing the availability of intracellular Ca2+, which is responsible for guanylate cyclase activation. Thus, dual pathways are proposed for a receptor-mediated cGMP rise in NG108-15 cells. PMID- 2986552 TI - Molybdate and the 1,25-dihydroxyvitamin D3 receptor from chick intestine. AB - The nature of the 1,25-dihydroxyvitamin D3 receptor from chick intestine was examined in regard to its response to sodium molybdate. Sodium molybdate (10 mM) stabilized the receptor from crude nuclear extract but not that from the supernatant or cytoplasmic fraction, suggesting the molybdate may act by binding to the DNA binding region of the receptor. At a concentration of 50 mM, sodium molybdate prevented aggregation of the nuclear receptor. This concentration of sodium molybdate also inhibited the receptor from binding to DNA cellulose while the same ionic strength KCl (90 mM) did not. These properties also suggest that molybdate interacts with the DNA binding region. Purification of the receptor using DNA cellulose chromatography has also been improved by using a sodium molybdate gradient (0-0.2 M) instead of the KCl gradient used previously. PMID- 2986553 TI - Isolation and characterization of 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase from bovine liver. AB - 6-Phosphofructo-2-kinase and fructose-2,6-bisphosphatase activities were copurified to homogeneity from bovine liver. The purification scheme consisted of polyethylene glycol precipitation, anion-exchange and Blue-Sepharose chromatography, substrate elution from phosphocellulose, and gel filtration. The bifunctional enzyme had an apparent molecular weight of 102,000 and consisted of two subunits (Mr 49,000). The kinase had a Km for ATP of 12 microM and a S0.5 for fructose 6-phosphate of 150 microM while the bisphosphatase had a Km for fructose 2,6-bisphosphate of 7 microM. Both activities were subject to modulation by various effectors. Inorganic phosphate stimulated both activities, while alpha glycerolphosphate inhibited the kinase and stimulated the bisphosphatase. The pH optimum for the 6-phosphofructo-2-kinase activity was 8.5, while the fructose-2,6 bisphosphatase reaction was maximal at pH 6.5. Incubation of the purified enzyme with [gamma-32P]ATP and the catalytic subunit of the cAMP-dependent protein kinase resulted in 32P incorporation to the extent of 0.7 mol/mol enzyme subunit with concomitant inhibition of the kinase activity and activation of the bisphosphatase activity. The mediation of the bisphosphatase reaction by a phosphoenzyme intermediate was suggested by the isolation of a stable labeled phosphoenzyme when the enzyme was incubated with fructose 2,6-[2 32P]bisphosphate. The pH dependence of hydrolysis of the phospho group suggested that it was linked to the N3 of a histidyl residue. The 6-phosphofructo-2 kinase/fructose-2,6-bisphosphatase from bovine liver has properties essentially identical to those of the rat liver enzyme, suggesting that hepatic fructose 2,6 bisphosphate metabolism is under the same control in both species. PMID- 2986554 TI - Role of sulfatide on phosphoenzyme formation and ouabain binding of the (Na+ + K+)ATPase. AB - A microsomal fraction rich in (Na+ + K+)ATPase activity has been isolated from the outer medulla of pig kidney. The ability of this preparation to form phosphoenzyme on incubation with [gamma-32P]ATP and to bind [3H]ouabain was studied when its sulfatide was hydrolyzed by arylsulfatase treatment. The K+ dependent hydrolysis of the Na+-dependent phosphorylated intermediate as well as the ouabain binding were inactivated in direct relation to the breakdown of sulfatide. Both characteristics of the (Na+ + K+)ATPase preparation, lost by arylsulfatase treatment, were partially restored by the sole addition of sulfatide. These experiments indicate that sulfatide may play a role in sodium ion transport either in the conformational transition of the K+-insensitive phosphointermediate, E1P, to the K+-sensitive intermediate, E2P, or in the configuration of the high-affinity binding site for K+ of the E2P form. In addition, this glycolipid may have a specific role in the proteolipidic subunit that binds ouabain. PMID- 2986555 TI - Phosphorolysis of acetyl phosphate by orthophosphate with energy conservation in the phosphoanhydride linkage of pyrophosphate. AB - The formation of pyrophosphate as a result of nucleophilic attack by orthophosphate at the acylphosphate bond of acetyl phosphate was detectable in completely aqueous media, and was enhanced by dimethyl sulfoxide. The reaction had an absolute requirement for divalent cations, the rate constant of phosphorolysis being dependent on the species and concentration of cations as well as on temperature and pH. The amount of pyrophosphate formed depended on both the acetyl phosphate and orthophosphate concentrations. In purely aqueous media, phosphorolysis was barely detectable in the presence of Mg2+, and its rate increased 40-fold when Mg2+ was replaced by Ca2+ or Sr2+. In the presence of Mg2+ the rate of phosphorolysis increased 400-fold when 50 to 80% of the water was replaced by dimethyl sulfoxide. In the latter case, the rate also increased as the pH was raised from 4.0 to 9.0. The entropy of activation was large and negative in the presence of Mg2+ or Ca2+, indicating that the nucleophile is involved in the rate-limiting step of the reaction. Since this thermodynamic parameter became large and positive in the presence of Ca2+ when dimethyl sulfoxide was omitted, it is inferred that the transition state of the same reaction may be changed by the solvent composition and the solvation of reactants. PMID- 2986556 TI - Superoxide-dependent lipid peroxidation and vitamin E content of microsomes from hepatomas with different growth rates. AB - Lipid peroxidation of microsomal membranes isolated from rat liver, and Morris hepatomas 9618A (slow-growing) and 3924A (fast-growing) was induced by superoxide radicals generated by the action of xanthine oxidase on xanthine. The peroxidation, measured as malondialdehyde and lipid hydroperoxide formation, was optimized with regard to iron concentration and chelation of iron by ADP. In such conditions hepatoma microsomes catalyze lower rates of lipid peroxidation than the normal counterpart. However, while microsomes from hepatoma 3924A show a marked decrease in both the malondialdehyde and hydroperoxide production rates, microsomes from hepatoma 9618A differ moderately from the control, mainly in the long-term production of hydroperoxides. It is also reported here that the 9618A microsomes partially lack cytochrome P-450 (about 40% deficiency), but they have a fatty acid composition similar to that of control. No differences were found in the content of vitamin E between normal and hepatoma 3924A microsomes. Moreover, induction of vitamin E deficiency in hepatoma 3924A microsomes does not influence the rate of either malondialdehyde or lipid hydroperoxide production. On the basis of these results and previous data on the lipid composition of hepatoma 3924A microsomes it is proposed that the high resistance to superoxide-dependent lipid peroxidation of hepatoma 3924A microsomes is related to the low substrate availability rather than the content of membrane antioxidants; and a limitation only in the propagation phase characterizes the hepatoma 9618A microsomal lipid peroxidation and would be due to the partial deficiency of the endogenous propagating agent, cytochrome P-450. PMID- 2986557 TI - A simple, rapid method for the preparation of plasma membranes from liver. AB - A method for the preparation of plasma membrane fractions from liver is described. A crude membrane fraction, substantially free of mitochondria and nuclei, is collected by centrifuging liver homogenate over a pad of 1.45 M sucrose. This interfacial fraction is further purified by isopycnic centrifugation in a self-generating density gradient of Percoll. All steps are carried out in buffered, isotonic solutions. Membrane prepared in this way is enriched in marker enzymes associated with plasma membrane, while marker enzymes for cellular components other than plasma membrane are diminished. Contamination by mitochondrial components is very much less than for membrane prepared from hypotonic liver homogenates. The membrane fraction also shows specific insulin binding, and negligible degradation of insulin. The procedure is rapid, uses simple equipment, and can be completed within 3 h. PMID- 2986559 TI - [Experience with induced hypertension chemotherapy for lung cancer]. AB - Induced hypertension chemotherapy (IHC) using angiotensin II was applied for patients with lung cancer who had not been treated previously, and the results compared with those of preceding conventional chemotherapy as a sequential control. Twenty-nine patients with non-small cell lung cancer (non-SCLC) were treated with MTX and MMC. Response rate among evaluable cases was 23.1% (3/13) for conventional chemotherapy and 18.2% (2/11) for IHC. Twenty-eight patients with small cell lung cancer (SCLC) were treated with VCR, CPA and ACNU. Among evaluable cases, both chemotherapy groups with and without IHC showed the same response rate, 66.7% (8/12). With respect to response rate, there were no differences between conventional chemotherapy and IHC for non-SCLC or SCLC. PMID- 2986560 TI - Vitamin D resistance and alopecia. A causal or casual relationship? PMID- 2986558 TI - [Preoperative chemo-embolization for treatment of hepatocellular carcinoma]. AB - A multidisciplinary treatment for hepatocellular carcinoma including preoperative chemo-embolization with resection treatment was introduced. Recent attempts at combining lipiodol with chemo-embolization have been quite promising. An ideal multidisciplinary treatment should include an effective device using the transportal approach. PMID- 2986561 TI - Calcitriol-resistant rickets with alopecia. AB - Four children of two kindred had alopecia associated with severe rickets, resistant to treatment, and caused by defective cytoplasmic and nuclear receptors for the active vitamin D metabolite calcitriol (1,25-dihydroxyvitamin D3). Scalp biopsy specimen revealed a normal number and light microscopic features of the hair and hair follicles. Calcitriol-resistant rickets should be added to the list of inherited disorders of hair growth, and the association of rickets with alopecia needs to be considered in the differential diagnosis of hair loss. PMID- 2986563 TI - Tumors of the adrenal cortex and Cushing's syndrome. AB - Fifty-nine patients with Cushing's syndrome, due to adrenocortical tumor, were studied and treated during the period 1953 through 1983 at Vanderbilt University Medical Center. Cushing's syndrome is caused by hypercortisolism that can be due to (1) medicinal use of steroids, (2) excess pituitary secretion of adrenocorticotropin (ACTH) (Cushing's disease), (3) adrenocortical tumor, benign or malignant, and (4) the ectopic ACTH syndrome. Clinical and endocrinologic features of Cushing's syndrome are described, and differential diagnosis of adrenocortical tumor by precise endocrinologic studies is detailed. Computerized axial tomographic (CAT) scan is currently the most accurate imaging modality for preoperative localization of tumors. Preoperative differential diagnosis between adrenocortical adenoma and carcinoma has become fairly accurate. Operative approaches in each category are described. Follow-up from 1 to 30 years has been completed for all patients, except for one who was lost after 7 years. Results of surgical treatment of adrenocortical adenomas are excellent, but the salvage from adrenocortical carcinomas is poor. PMID- 2986562 TI - Does preoperative percutaneous biliary drainage reduce operative risk or increase hospital cost? AB - Despite recent advances in perioperative support care, surgery for obstructive jaundice is still associated with significant morbidity and mortality. For this reason, preoperative percutaneous transhepatic drainage (PTD) has been recommended for these patients. This method of management, however, has only been supported by retrospective and nonrandomized studies. Therefore, a prospective, randomized study was performed to determine the effect of preoperative PTD on operative mortality, morbidity, hospital stay, and hospital cost. Thirty-day mortality was 8.1% among 37 patients undergoing preoperative PTD, compared to 5.3% for 38 patients who went to surgery without preoperative drainage. Overall morbidity was also slightly, but not significantly, higher in patients who underwent preoperative PTD, (57% versus 53%). However, total hospital stay was significantly longer (p less than 0.005) in the PTD group (31.4 days versus 23.1 days). The cost of this excess hospitalization and the PTD procedure at our university medical center was over +8000 per patient. The authors conclude that preoperative PTD does not reduce operative risk but does increase hospital cost and, therefore, should not be performed routinely. PMID- 2986566 TI - The heart. An endocrine organ (revisited). PMID- 2986565 TI - A multifactorial analysis of steroid hormone receptors in stages I and II breast cancer. AB - It has been shown that the level of estrogen receptors (ER), and to some extent progesterone receptors (PR), correlate to a high degree to the response to endocrine therapy in advanced breast cancer patients. To evaluate the prognostic value of ER/PR in early breast cancer, 80 patients with stages I and II were studied. They all underwent modified radical mastectomy. Patients with stage I disease (negative LN) received no further treatment, while those with stage II received standard adjuvant chemotherapy. All the patients were followed for 4 years. The ER and PR were measured in each primary tumor by the glycerol density gradient method. Values of 10 fmole/mgm protein or greater were considered positive (+) and less than 10 fmole/mgm were considered negative (-). The results revealed: (1) Fifty-two patients (65%) had ER+, of which 44 (85%) were also PR+; 28 patients had ER-, of which 24 were also PR- (p less than 0.0001). (2) ER/PR correlated with age as 71% of the patients over age 50 had ER+/PR+, compared to 33% of those under age 50 (p less than 0.05). (3) Postmenopausal patients had a higher incidence of ER+/PR+. (4) Primary tumors less than 2 cm in size had higher ER+; 71% in those with stage I and 80% in stage II. (5) Fifty-eight per cent (38) of patients with ductal carcinoma had ER+/PR+, compared to 67% (4) with lobular carcinoma. (6) The disease-free survival of patients with ER+ tumors was significantly longer than those with ER- tumors (p less than 0.005) both in positive and negative LN patients. The same was true for PR+ compared to PR- (p less than 0.005), but only in those with stage II disease. The overall survival rates were similarly significant in favor of ER+ and PR+ patients (p less than 0.025), but only in stage II disease. It seems that the status of steroid hormone receptors has a major prognostic factor second only to the LN status. PMID- 2986564 TI - Cushing's disease today. Late follow-up of 17 adrenalectomy patients with emphasis on eight with adrenal autotransplants. AB - Cushing's disease has come full cycle. As originally asserted more than 50 years ago, modern diagnostic techniques now demonstrate an adrenocorticotropic hormone (ACTH) secreting pituitary adenoma in approximately 80% of such patients. At this historical juncture, we report a long-term follow-up of our 17 patients who underwent adrenalectomy (8) or later adrenalectomy plus adrenal autotransplantation (9) between 1955 and 1976. Two patients died soon after surgery and five others died later of "natural" causes. Four others moved away but were stable when last contacted. Of the six patients who remain available for current follow-up, three have undergone hypophyseal surgery. Another patient has evidence of pituitary enlargement, and the remaining two are yet to undergo computerized tomography (CT) scan. Four illustrative cases are reviewed in some detail. One case presented with Nelson's syndrome and acute onset blindness. The second represented multiple endocrine adenomatosis with hyperparathyroidism in addition to Cushing's disease. The third exhibited Cushing's syndrome from the autotransplants, finally cured by hypophysectomy. The fourth exhibited huge ACTH levels from a large pituitary adenoma that could not be totally resected and recurrent Cushing's syndrome associated with large autotransplant "adenomas." The initial surgical treatment of choice is pituitary adenectomy. Bilateral adrenalectomy will remain useful where curative pituitary surgery is not feasible. Neither pituitary irradiation nor medical therapy has been truly effective in our patients. Adrenal autotransplants survive, to some extent, in virtually all patients. However, the degree of function is variable, and the full function may not be achieved for many months or even years. Functioning autotransplants have not prevented Nelson's syndrome, and they would appear to offer little practical benefit at this time. PMID- 2986567 TI - Ketoconazole. Use in the treatment of ectopic adrenocorticotropic hormone production and Cushing's syndrome in small-cell lung cancer. AB - Recently, ketoconazole, an imidazole derivative with antifungal properties, has been shown to inhibit adrenal corticosteroid and androgen production. We treated a patient with small-cell lung cancer and Cushing's syndrome secondary to ectopic secretion of adrenocorticotropic hormone in whom it was necessary to discontinue chemotherapy. The patient was treated with ketoconazole, with a resultant reduction in urinary free cortisol and ketosteroid excretion and improvement in electrolyte balance. Ketoconazole may be a useful adjunct in the treatment of small-cell lung cancer or other malignancies associated with excessive corticosteroid production or as a hormonal therapy in breast or prostatic cancer. PMID- 2986568 TI - A simple and rapid method for screening bacteria for type II restriction endonucleases: enzymes in Aphanothece halophytica. AB - A method is described which allows a large number of bacterial strains to be rapidly and easily screened for the presence of site-specific endonucleases. The method involves selective permeabilization of the bacterial cell and analysis of the exuded material. Type II restriction endonucleases from cyanobacteria and Gram-negative eubacteria have been detected and new enzymes have been found. The method should be widely applicable and easy to modify for use in genera other than those tested. Three-site-specific endonuclease activities, detected by this method in Aphanothece halophytica PCC 7412, were purified and their recognition and cleavage specificities were determined AhaI and AhaII recognise and cleave the same DNA sequences as CauII and AcyI respectively; the specificity of AhaIII (TTT decreases AAA) has been reported previously (Whitehead and Brown, 1982, FEBS Letters 143:296-300). PMID- 2986569 TI - The plasmids of Deinococcus spp. and the cloning and restriction mapping of the D. radiophilus plasmid pUE1. AB - Plasmids were found in strains representing all four species of the genus Deinococcus viz. D. radiodurans, D. radiopugnans, D. radiophilus and D. proteolyticus but were not found in the most intensively-investigated strain of the genus, D. radiodurans R1. Their sizes were calculated from electron micrographs. D. radiophilus yielded three size classes of plasmid while D. radiodurans Sark, D. proteolyticus and D. radiopugnans each yielded two. Attempts to cure D. radiophilus and D. radiodurans Sark of any of their plasmids, using a variety of methods, were unsuccessful. A 10.8 kbase pair (kb) plasmid from D. radiophilus, pUE1, was cloned into the PstI site of pAT153 and propagated in Escherichia coli HB101. The recombinant plasmid, pUE109 was subjected to single and double digestion with various restriction endonucleases and its restriction map constructed. The resistance of E. coli HB101 to ultraviolet radiation was not increased when pUE109 was introduced into it. Attempts to transform D. radiodurans with pUE109 failed to detect tetracycline-resistant transformants. PMID- 2986570 TI - [Reactivity of sheep to the application of plasma membranes from cattle lymph node tumors]. PMID- 2986571 TI - Pituitary gland in hypothyroidism. Histologic and immunocytologic study. AB - Primary hypothyroidism is associated with hypertrophy and hyperplasia of thyrotropic cells. In addition, pituitary adenomas that produce thyroid stimulating hormone occur in both hypothyroidism and hyperthyroidism. The relationship between thyrotropic hyperplasia and adenoma formation is, however, unsettled. We summarize the results of a histologic and immunocytologic study of the pituitary glands of 64 patients with long-standing primary hypothyroidism in an effort to characterize the changes in thyrotropic cells as related to the duration and severity of disease, to therapy, and to the development of thyrotropic adenomas. Diffuse and nodular thyrotropic cell hyperplasia was noted in 69% and 25% of glands, respectively. A crude correlation was observed between the degree of thyrotropic cell hyperplasia and the relative lack of thyroid hormone replacement therapy. In 12% of glands, tumorlet formation was observed, perhaps representing an intermediate stage between nodular hyperplasia and the development of microadenoma. Twelve adenomas were noted, five of which contained thyroid-stimulating hormone immuno-reactive cells. Although thyroid hormone deficiency seemed to selectively affect thyrotropic cells, lactotropic hyperplasia was observed in 20% of patients; the mechanism accounting for prolactin cell hyperplasia remains obscure. PMID- 2986572 TI - Immunohistochemical localization of chromogranin in human hypophyses and pituitary adenomas. AB - Chromogranin was demonstrated by immunohistochemistry in the cytoplasm of human beta-thyrotropin, human beta-follicle-stimulating hormone-, human beta luteinizing hormone-, and human alpha-subunit-containing cells of the non tumorous human adenohypophyses. Some surgically removed human beta-thyrotropin-, human beta-follicle-stimulating hormone-, human beta-luteinzing hormone-, and human alpha-subunit-producing pituitary adenomas, as well as some null cell adenomas, exhibited chromogranin immunoreactivity, whereas adenomas storing human growth hormone, human prolactin, or corticotropin were negative. Chromogranin immunopositivity was variable in extent and intensity; not every glycoprotein producing cell could be immunostained in the nontumorous adenohypophysis and the majority of chromogranin-containing adenomas showed only focal positivity. No explanation can be offered for this variability. The demonstration of chromogranin by the avidin-biotin-peroxidase technique may be helpful in the immunohistochemical characterization of some glycoprotein hormone-producing pituitary adenomas, as well as null-cell adenomas of the human pituitary. PMID- 2986574 TI - Angiomatoid malignant fibrous histiocytoma. Cytology of fine-needle aspiration and its differential diagnosis. AB - A case of angiomatoid malignant fibrous histiocytoma was studied by fine-needle aspiration biopsy and routine histologic methods. Smears from aspirated material consisted of numerous large loose cellular aggregates with occasional single cells. Histiocytelike cells and lymphocytes were the prominent cell types with lesser numbers of spindle-shaped, fibroblastlike cells. Moderate cytologic atypia was noted. A cystic mass with the above cytologic findings occurring in the superficial soft tissue of the extremity of a young patient should suggest the diagnosis of an angiomatoid malignant fibrous histiocytoma. PMID- 2986573 TI - Necrotizing lymphoid vasculitis in X-linked lymphoproliferative syndrome. AB - An 8-year-old maternally related relative of three boys who had developed agammaglobulinemia associated with Epstein-Barr virus (EBV)-induced infectious mononucleosis was studied for X-linked lymphoproliferative syndrome (XLP) in 1979. At that time, he demonstrated no striking immunologic aberrations and was seronegative for EBV. Subsequently, immunologic abnormalities including failure to switch from IgM to IgG antibody synthesis after secondary immunization with bacteriophage phi X174 were detected. In 1983, he experienced episodic intracerebral hemorrhages, with the second being fatal. At autopsy, necrotizing vasculitis and aneurysms involving arteries of the central nervous system were observed. Studies of blood obtained immediately before and after death failed to show antibodies to EBV. However, EBV genome was demonstrated in tissues obtained at autopsy by DNA hybridization studies. Fatal lymphoid vasculitis in this patient is unique among boys with XLP in the registry. These findings probably extend the phenotypic expressions of XLP. PMID- 2986575 TI - Effects of chlorpromazine and its dihydroxy metabolite on calcium movement and electrical activity in superior cervical ganglia. AB - Chlorpromazine (CPZ) binding and the binding to isolated guinea pig superior cervical of a toxic metabolite 7,8-dihydroxychlorpromazine (7,8-diOH-CPZ) were studied under resting and potassium-stimulated conditions. Maximal binding occurred more rapidly in high potassium, with binding of CPZ 10 fold higher than that of 7,8-diOH-CPZ. However, in low potassium media 50% of bound CPZ and 60% of 7,8-diOH-CPZ was retained following a 240-min washout procedure. When the washout of the drugs from the tissue was studied, release was found to be more extensive in high potassium washout media. Under resting conditions, at 10 microM CPZ or 7,8-diOH-CPZ, calcium accumulation by ganglia decreased but increased at higher drug concentrations. Calcium (Ca) uptake increased with both drugs in high potassium medium. Ca efflux from the tissue was accelerated by both phenothiazines in a dose-dependent fashion. CPZ blocked ganglionic transmission at 20 microM while 7,8-diOH-CPZ (up to 100 microM) increased ganglionic potential amplitude. Thus CPZ and 7,8-diOH-CPZ differ in binding to ganglia and in effects on ganglionic transmission. PMID- 2986576 TI - Characterization of a new calicivirus isolated from feces of a dog. AB - Canine calicivirus (CaCV), isolated from feces of a dog with diarrhea, was readily propagated in cultures of canine cells and in a dolphin cell line. Serologic evidence indicated many dogs in at least one geographic area had been infected with CaCV, but its role as an etiologic agent of disease was not established. In cell culture most CaCV virions were strongly cell-associated making purification difficult. CaCV was established as a member of the Caliciviridae by morphology and physicochemical properties of virions (density, sedimentation rate, single major polypeptide, RNA genome size), although some of the properties differed slightly from those of previously described caliciviruses; evidence was also obtained for caliciviral RNA species in infected cells. Based on tests with antisera to numerous caliciviruses and presumed caliciviruses, CaCV appeared to be not closely related to any previously described virus except the stunting syndrome agent of chickens. PMID- 2986577 TI - Gel electrophoretic analysis of polypeptides from nucleocapsids of Marek's disease virus strains and herpesvirus of turkey. AB - The polypeptides of nucleocapsids of Marek's disease virus (MDV) strains with different biological properties and of antigenically related herpesvirus of turkey (HVT) strains were analysed by one- and two-dimensional (1D and 2D, respectively) gel electrophoresis. Based on small differences in migration behaviour (size and charge) of a number of corresponding nucleocapsid polypeptides, the virus strains could be differentiated into three groups. The polypeptide pattern of group I, comprising the virulent MDV-strain K and the attenuated strains, HPRS-16/att and CVI988 37th passage, was composed of four major polypeptides (i.e. 140K, 50K, 40K and 33K daltons) and at least four minor polypeptides. The pattern of group II, comprising the naturally occurring non oncogenic MDV-strains SB-1 and HPRS-24, contained one additional major polypeptide of 39K daltons. The nucleocapsid-specific 2D polypeptide patterns of the HVT strains HVT-Fc 126 and PB-THV1, comprising group III, were distinguishable from each other on the basis of a small difference in size of one major 50K polypeptide. Results were further substantiated by coelectrophoresis experiments. PMID- 2986578 TI - Detection of human polyomavirus DNA in urine specimens by hybridot assay. AB - A hybridot assay method using a labelled BK virus probe has been used to detect the presence of human polyomavirus DNA in 81 urine specimens from 61 patients, most of whom were immuno-compromised to some degree. Twenty-eight urines from 23 patients had detectable DNA. The results have been compared to virus isolation and electron microscopy on the same specimens. In 90 per cent a comparable result was obtained by at least one of the other two tests and in the 73 specimens on which all 3 tests were performed there was 80 per cent agreement between all. In 6 cases the hybridot assay was more sensitive in detecting infection. PMID- 2986580 TI - Parvovirus (feline panleucopaenia virus) plaque formation. AB - A plaque assay was developed for feline parvovirus (FPV; feline panleucopaenia virus) in a feline embryo (FEmb) cell line. Higher numbers and larger diameter plaques were obtained with a) seeding rates of 0.7 X 10(5) and 1.5 X 10(5) cells cf. 3 X 10(5) and 6 X 10(5) cells/well of 35 mm diameter, b) synchronised cells infected at the G1-S interface cf. nonsynchronised cells and c) 5 to 6 days incubation post inoculation. The plaque assay was standardised by using serum deprivation for 24 hours to synchronize cells, a seeding rate of 1.5 X 10(5) cells/35 mm diameter well, inoculation of virus 16 hours post seeding followed by 5 days incubation. The standardised assay gave consistent, reproducible results. A dose-response curve using the assay showed a linear, 45 degrees slope, relationship between plaque forming units and virus dilution which further verified the sensitivity and reliability of the assay. Plaques produced by "wild" type and plaque purified virus were invariably non uniform in diameter; diameter of plaques in fact followed a normal frequency distribution under standard assay conditions. PMID- 2986579 TI - An enteric coronavirus of the rabbit: detection by immunoelectron microscopy and identification of structural polypeptides. AB - The immunoelectron microscopy (IEM) technique has been used for the detection of a rabbit enteric coronavirus (RECV). Immune serum was prepared in guinea pigs; the viral antigen used for the immunization procedure was obtained from the caecum of a sick rabbit, concentrated by centrifugation and purified on Percoll gradient. In order to identify the viral particles used in the immunization procedure, the protein pattern of the particles was determined by electrophoresis and compared with the pattern of other known coronaviruses. Analysis of structural polypeptides of the purified viral particles revealed a pattern similar to that reported for other coronaviruses. These polypeptides cross reacted with two other coronavirus specific immune sera (IBV and TGE). IEM assay of fecal samples collected from healthy and sick rabbits showed the presence of immune aggregates in specimens from both sick and healthy rabbits. Those aggregates contained viral particles sharing morphological characteristics with other coronaviruses. Furthermore, IEM assay was shown to be more sensitive than a direct EM procedure to detect coronavirus particles in rabbit feces. This assay also allowed the detection of a larger number of chronic carriers. PMID- 2986581 TI - Effect of salts and other agents on foot-and-mouth disease virus poly (U) polymerase activity. AB - The activity of the purified poly(U) polymerase replication complex of foot-and mouth disease virus was optimized when 100 mM NH4+ and either 0.75 mM Al3+ or 1.0 mM Fe3+ was added to the standard assay reaction mixture. Zn2+ at concentrations of 10(-5) mM to 5 mM inhibited enzyme activity although all polymerases examined to date have contained zinc. Mercaptoethanol and dithiothreitol inhibited polymerase activity despite the presence of cysteine residues in the viral induced polypeptide of the replication complex, possibly because of their action as metal chelators rather than as reducing agents. PMID- 2986584 TI - [Circulatory status and architectonics of the vascular bed of the leg after lengthening it following double-distraction epiphysiolysis]. AB - In 6-7-month-old puppies of mongrel dogs under anesthesia by means of G. A. Ilizarov apparatus double epiphysiolysis of the tibia has been performed. The activity of metabolic processes, the state of blood and lymphatic circulation have been studied in the shin at its lengthening by radioisotopic, angiographic and histological methods. Beginning from the first postoperative days, an essential increase of metabolic processes is observed in the osseous tissue, in the blood circulation at the areas of distraction and in the shin as a whole. Blood supply of the regenerates, when the magistral and intraosseous blood stream is remained intact, is performed via the nutritional, metaphysial and epiphyseal arteries. The preservation of the magistral blood stream ensures formation of a normal blood bed in the red bone marrow (arterioles, closed system of the sinuses). PMID- 2986582 TI - Comparison of contagious ecthyma virus genomes by restriction endonucleases. AB - Restriction enzyme analysis with KpnI revealed heterogeneity among 10 different strains of contagious ecthyma virus from sheep, musk ox, Dall sheep and humans. Hybridization of electroblots indicated deletion of DNA segments and changes in nucleotide sequences. PMID- 2986583 TI - Properties of temperature-sensitive mutants of parainfluenza virus type 3 selected during the course of persistent infection. AB - We investigated properties of the ts mutants that were selected during the course of persistent infection of Vero cells by parainfluenza virus type 3. The mutants demonstrated leakiness when infecting cells at high MOI and interfered with the growth of wild type virus, apparently by inhibiting a step prior to RNA synthesis. PMID- 2986586 TI - The influence of the sympathetic nervous system on the development of beta adrenergic receptors in the rat submandibular salivary gland. AB - During the development of the rat submandibular gland (SMG) there is a clearly defined sequence in the maturation of the beta-adrenergic receptor/adenylate cyclase-linked stimulus-secretion coupling system. The sympathetic nervous system does not become functionally linked to the exocrine process in the SMG until approximately six days after birth. The temporal correlation of the ingrowth of catecholamine-containing nerve processes, the appearance of beta-adrenergic receptors and the functional coupling of the stimulus-secretion system suggested the possibility of a cause and effect relationship between the appearance of the catecholamine-containing nerves in the gland and the maturational increase in the number of beta-adrenergic receptors. Chemical sympathectomy in neonates did not effect the time of appearance or the number of beta-adrenergic receptors seen in the developing gland. However, chronic isoproterenol treatment resulted in accelerated maturation of the gland with a concomitant premature appearance of the beta-receptors. These data suggest that the increase in the number of beta adrenergic receptors which normally occurs in the developing gland at 5-6 days after birth is a specifically-programmed step closely associated with the degree of maturation attained by the cells and is independent from the ingrowth of catecholamine-containing nerves. PMID- 2986585 TI - Effect of time, degree of saturation, pH and acid concentration of buffer solutions on the rate of in-vitro demineralization of human enamel. AB - The rate of demineralization of the artificial caries-like process was measured using two methods. The rate calculated from microradiography of the demineralized enamel appeared to be equal to the one calculated from calcium and phosphate analysis of the demineralizing buffer. In small volumes of demineralizing solution, the rate of demineralization decreased with time, due to the increase in saturation of the solution during the demineralization of the enamel. With increasing saturation, with respect to hydroxyapatite, and with increasing pH the rate of demineralization decreased. Increasing acid concentration had a marked influence only at the lower concentrations; a further increase had no effect on the rate of demineralization. PMID- 2986587 TI - Characterization of a human hepatic receptor for high density lipoproteins. AB - Characterization of the membrane receptor for the low density lipoproteins (LDL) has led to insights into cellular receptor physiology as well as mammalian lipid transport. Result with LDL have stimulated the search for specific receptors for other plasma lipoproteins. Receptors for high density lipoproteins (HDL) have been identified in human fibroblasts and smooth muscle cells. Specificity for this receptor has been difficult to define since normal HDL contains several apolipoproteins, and particles containing apolipoproteins B and E have been shown to compete for HDL binding. In the present study, we demonstrate that HDL isolated from a patient devoid of apolipoprotein E was bound specifically by human hepatic membranes. This binding reached saturation within 2 hours and was EDTA-resistant. Assuming a single receptor model, we found that 2.9 x 10(15) receptors/mg membrane protein bound with an affinity KD = 3.5 x 10(-7) M at 0 to 4 degrees C and KD = 1.9 x 10(-7) M at 37 degrees C. The binding was effectively competed with intact HDL3, with HDL3 that had undergone selective arginine and lysine residue modification, and with antibodies to apolipoproteins A-I and A-II. However, LDL, asialofetuin, and HDL3 which had undergone tyrosine modification by nitration, and anti-apolipoprotein B did not compete with apo A-I HDL binding. In contrast to LDL binding, the human hepatoma cell line, HEPG2, increased HDL binding with cholesterol loading that was specific for HDL3. Thus, hepatic tissue can modulate its recognition of HDL. Finally, hepatic membranes from a patient lacking normal hepatic LDL receptors bound apo A-I HDL normally. These data indicate that a saturable, specific regulatable receptor for apo E-free HDL is present in human liver. PMID- 2986588 TI - ACTH4-10 produces a transient decrease in septal hyperemotionality. AB - The administration of doses of ACTH4-10 that have been shown to be effective in producing enhanced peripheral nerve regeneration did not produce the enhanced rate of decline in hyperemotionality produced by administration of the ACTH4-9 variant Org 2766. Long-term changes in septal reactions in training situations were likewise not affected by ACTH4-10. These results indicate that, despite similar peripheral nerve growth effects, these two ACTH-related agents (ACTH4-10 and Org 2766) produce quite different long- and short-term effects on rats with septal area damage. PMID- 2986589 TI - Effect of variations in the conditions of DNA synthesis upon the accuracy of DNA replication. PMID- 2986591 TI - Non-invasive measurements of cell surface receptors. AB - None of the current methods for assessing the effects of subtle changes in drug structure on tissue is entirely satisfactory. The observation that 3H-drug molecules emit photons of light when they bind to membrane proteins (called Condensed Phase Radioluminescence) probably represents a significant advance in measuring non-invasively kinetic effects of drugs on beating monolayers of cardiac myocytes. The potential utility of CPR is great. PMID- 2986590 TI - Enzymatic channeling of DNA precursors. PMID- 2986592 TI - [Simultaneous vaccination against Aujeszky's disease and European hog cholera]. PMID- 2986593 TI - Polymorphism of mitochondrial DNA in pigs based on restriction endonuclease cleavage patterns. AB - Restriction endonuclease cleavage patterns of mitochondrial DNA (mtDNA) of pigs and Japanese wild boars were analyzed using 17 enzymes which recognize six nucleotides. The map of cleavage sites was made by double-digestion methods. Polymophism of mtDNA was detected in the digestion by BglII, EcoRV, ScaI, and StuI. The restriction cleavage patterns were identical among the breeds of Landrace, Hampshire, Duroc I, and Large White I (A type). The patterns of Large White II were the same as those of Japanese wild boars (B type). A difference between the A type and the B type of mtDNA was found in the case of three restriction enzymes, BglII, ScaI, and StuI, and the nucleotide alterations between them were estimated as more than six. On the other hand, a difference between mtDNA from almost all pigs and mtDNA from Duroc II was detected using EcoRV. We suggest that the difference of mtDNA between the A type and the B type of mtDNA could result from the different origin of boars, that is, whether they were of European or Asian origin. PMID- 2986594 TI - Physicochemical behaviour and structural characteristics of membrane-bound acetylcholinesterase from Torpedo electric organ. Effect of phosphatidylinositol specific phospholipase C. AB - Quantitative solubilization of the phospholipid-associated form of acetylcholinesterase (AChE) from Torpedo electric organ can be achieved in the absence of detergent by treatment with phosphatidylinositol-specific phospholipase C (PIPLC) from Staphylococcus aureus [Futerman, Low & Silman (1983) Neurosci. Lett. 40, 85-89]. The sedimentation coefficient on sucrose gradients of AChE solubilized in detergents (DSAChE) varies with the detergent employed. However, the coefficient of AChE directly solubilized by PIPLC is not changed by detergents. Furthermore, PIPLC can abolish the detergent-sensitivity of the sedimentation coefficient of DSAChE purified by affinity chromatography, suggesting that one or more molecules of phosphatidylinositol (PI) are co solubilized with DSAChE and remain attached throughout purification. DSAChE binds to phospholipid liposomes, whereas PIPLC-solubilized AChE and DSAChE treated with PIPLC do not bind even to liposomes containing PI. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis shows that PIPLC-solubilized AChE, like unmodified DSAChE, is a catalytic subunit dimer; electrophoresis in the presence of reducing agent reveals no detectable difference in the Mr of the catalytic subunit of unmodified DSAChE, of AChE solubilized by PIPLC and of AChE solubilized by Proteinase K. The results presented suggest that DSAChE is anchored to the plasma membrane by one or more PI molecules which are tightly attached to a short amino acid sequence at one end of the catalytic subunit polypeptide. PMID- 2986596 TI - Inhibitors of angiotensin-converting enzyme containing a tetrahedral arsenic atom. AB - A series of tetrahedral oxo acids of Group VA and VIA elements and of silicon and boron were examined as inhibitors of angiotensin-converting enzyme. Arsenate is a competitive inhibitor with a Ki of 27 +/- 1 mM, at least 10-fold more potent than phosphate. Dimethylarsinate is a competitive inhibitor with a Ki of 70 +/- 9 mM, 2-fold more potent than dimethylphosphinate. Oxo acids of boron, silicon, antimony, sulphur and selenium are not inhibitors. On the basis of these results and the strong inhibition of this zinc metallopeptidase by substrate analogues containing a tetrahedral phosphorus atom, two substrate analogues containing a tetrahedral arsenic atom were prepared. 2-Arsonoacetyl-L-proline is a competitive inhibitor with a Ki of 18 +/- 7 mM, more than 2000-fold weaker than that of its phosphorus analogue 2-phosphonoacetyl-L-proline. 4-Arsono-2-benzylbutanoic acid is a mixed inhibitor with a Ki of 0.5 +/- 0.2 mM, indistinguishable in potency from its phosphorus analogue 2-benzyl-4-phosphonobutanoic acid. PMID- 2986595 TI - Topography, purification and characterization of thyroidal NAD+ glycohydrolase. AB - Subcellular fractionation of bovine thyroid tissue by differential pelleting and isopycnic gradient centrifugation in a zonal rotor indicated that NAD(+) glycohydrolase is predominantly located and rather uniformly distributed in the plasma membrane. Comparison of NAD(+) glycohydrolase activities of intact thyroid tissue slices, functional rat thyroid cells in culture (FRT(l)) and their respective homogenates indicated that most if not all of the enzyme (catalytic site) is accessible to extracellular NAD(+). The reaction product nicotinamide was predominantly recovered from the extracellular medium. The diazonium salt of sulphanilic acid, not penetrating into intact cells, was able to decrease the activity of intact thyroid tissue slices to the same extent as in the homogenate. Under the same conditions this reagent almost completely abolished NAD(+) glycohydrolase activity associated with intact thyroid cells in culture. The triazine dye Cibacron Blue F3GA and its high-M(r) derivative Blue Dextran respectively completely eliminated or caused a severe depression in the NAD(+) glycohydrolase activity of FRT(l) cells. The enzyme could be readily solubilized from bovine thyroid membranes by detergent extraction, and was further purified by gel filtration and affinity chromatography on Blue Sepharose CL-6B. The overall procedure resulted in a 1940-fold purification (specific activity 77.6mumol of nicotinamide released/h per mg). The purified enzyme displays a K(m) of 0.40mm for beta-NAD(+), a broad pH optimum around pH7.2 (0.1 m-potassium phosphate buffer) and an apparent M(r) of 120000. Nicotinamide is an inhibitor (K(i) 1.9mm) of the non-competitive type. The second reaction product ADP-ribose acts as a competitive inhibitor (K(i) 2.7mm). The purified enzyme splits beta NAD(+), beta-NADP(+), beta-NADH and alpha-NAD(+) at rates in the relative proportions 1:0.75:<0.02:<0.02 and exhibits transglycosidase (pyridine-base exchange) activity. Anionic phospholipids such as phosphatidylinositol and phosphatidylserine inhibit the partially purified enzyme. A stimulating effect was observed upon the addition of histones. PMID- 2986597 TI - Free hydroxyl radical is not involved in an important reaction of lignin degradation by Phanerochaete chrysosporium Burds. AB - Hydroxyl radical (HO.) has been implicated in the degradation of lignin by Phanerochaete chrysosporium. This study assessed the possible involvement of HO. in degradation of lignin substructural models by intact cultures and by an extracellular ligninase isolated from the cultures. Two non-phenolic lignin model compounds [aryl-C(alpha)HOH-C(beta)HR-C(gamma)H2OH, in which R = aryl (beta-1) or R = O-aryl (beta-O-4)] were degraded by cultures, by the purified ligninase, and by Fenton's reagent (H2O2 + Fe2+), which generates HO.. The ligninase and the cultures formed similar products, derived via an initial cleavage between C(alpha) and C(beta) (known to be an important biodegradative reaction), indicating that the ligninase is responsible for model degradation in cultures. Products from the Fenton degradation were mainly polar phenolics that exhibited little similarity to those from the biological systems. Mass-spectral analysis, however, revealed traces of the same products in the Fenton reaction as seen in the biological reactions; even so, an 18O2-incorporation study showed that the mechanism of formation differed. E.s.r. spectroscopy with a spin-trapping agent readily detected HO. in the Fenton system, but indicated that no HO. is formed during ligninase catalysis. We conclude, therefore that HO. is not involved in fungal C(alpha)-C(beta) cleavage in the beta-1 and beta-O-4 models and, by extension, in the same reaction in lignin. PMID- 2986599 TI - Gonadotropin releasing hormone stimulates the formation of inositol phosphates in rat anterior pituitary tissue. AB - The production of inositol phosphates in response to gonadotropin releasing hormone (GnRH) was studied in rat anterior pituitary tissue preincubated with [3H]inositol. Prelabelled paired hemipituitaries from prepubertal female rats were incubated in the presence or absence of GnRH in medium containing 10 mM-Li+ X Li+, which inhibits myo-inositol-1-phosphatase, greatly amplified the stimulation of inositol phosphate production by GnRH (10(-7) M) to 159, 198 and 313% of paired control values for inositol 1-phosphate, inositol bisphosphate and inositol trisphosphate respectively after 20 min. The percentage distribution of [3H]inositol within the phosphoinositides was 91.3, 6.3 and 2.4 for phosphatidylinositol, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate respectively and was unaffected by GnRH. The stimulation of inositol trisphosphate production by GnRH was evident after 5 min incubation, was dose-dependent with a half-maximal effect around 11 nM, and was not inhibited by removal of extracellular Ca2+. Elevation of cytosolic Ca2+ by membrane depolarization with 50 mM-K+ had no significant effect on inositol phosphate production. These findings are consistent with the hypothesis that GnRH action in the anterior pituitary involves the hydrolysis of phosphatidylinositol 4,5 bisphosphate. The resulting elevation of inositol trisphosphate may in turn lead to intracellular Ca2+ mobilization and subsequent stimulation of gonadotropin secretion. PMID- 2986598 TI - Assembly of chick and bovine lens-capsule collagen. AB - Chick-embryo and adult bovine lens-capsular epithelia in organ culture synthesized 4-hydroxy[3H]proline-containing polypeptides when incubated in the presence of [3H]proline. These collagenous polypeptides of apparent Mr 180 000, 175 000 and 160 000 became incorporated with time into aggregates of higher molecular size. The formation of such aggregates was inhibited when the tissues were labelled in the presence of beta-aminopropionitrile, thereby implicating lysine-derived cross-links in aggregate formation. When the tissues were incubated in the presence of tunicamycin, the collagenous polypeptides synthesized exhibited increased electrophoretic mobilities on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The addition to lens-capsule incubation medium of alpha alpha'-bipyridine led to the synthesis of underhydroxylated type IV collagen, also of increased electrophoretic mobility. Extended pulse-chase experiments indicated that such underhydroxylated collagen did not participate in aggregate formation, but was at least as stable as fully hydroxylated non-cross-linked collagen synthesized in the presence of beta aminopropionitrile. Native type IV collagen, recovered from the culture medium when capsules were incubated with [3H]proline for 24h, was purified by ion exchange chromatography. Separations conducted on CM-cellulose under denaturing and nondenaturing conditions suggested that the alpha 1(IV) and alpha 2(IV) chains occur in the same heterologous triple helix. Densitometric analyses of appropriate fluorograms indicated that these two polypeptides occur in a 2:1 ratio, suggesting that lens-capsule collagen is synthesized as a triple-helical molecule of composition [alpha 1(IV)]2 alpha 2(IV). PMID- 2986600 TI - Studies on the mechanism of inhibition of glucose-stimulated insulin secretion by noradrenaline in rat islets of Langerhans. AB - Noradrenaline (norepinephrine) was shown to be a potent inhibitor of glucose induced insulin release from rat pancreatic islets, with half-maximal inhibition of the secretory response to 20 mM-glucose occurring at approx. 0.3 microM, and complete suppression of the response occurring at 4 microM-noradrenaline. Inhibition of insulin secretion by noradrenaline was antagonized by the alpha 2 adrenergic antagonist yohimbine (half maximally effective dose approximately 1 microM), but was largely unaffected by the alpha 1-adrenergic antagonist prazosin at concentrations up to 50 microM, suggesting that the response was mediated by alpha 2-adrenergic receptors. Noradrenaline significantly reduced the extent of 45Ca2+ accumulation in glucose-stimulated islets, although as much as 5 microM noradrenaline was required for 50% inhibition of this response. The ability of noradrenaline to inhibit islet-cell 45Ca2+ uptake was totally abolished in media containing 1 mM-dibutyryl cyclic AMP, suggesting that the response may have been secondary to lowering of islet cyclic AMP. Under these conditions, however, noradrenaline was still able to inhibit insulin secretion maximally. The data suggest that the site(s) at which noradrenaline acts to mediate inhibition of insulin secretion in rat islets lies distal to both islet-cell cyclic AMP accumulation and Ca2+ uptake. PMID- 2986602 TI - New insights on the cytochrome c oxidase proton pump. AB - Cytochrome c oxidase vesicles were used to show that, under appropriate experimental conditions: (1) no net deprotonation of the vesicular membrane or of the incorporated enzyme occurs during the oxidation of ferrocytochrome c; (2) the pH equilibration kinetics of a respiration-induced pH gradient across the bilayer are a simple function of the ohmic proton-conductance properties of the membrane; (3) a fairly constant stoichiometry (0.8-0.7) of the numbers of protons pumped per molecule of ferrocytochrome c oxidized, i.e. the H+/e- ratio, over a wide range of dioxygen molecules reduced (1-12) is observed. PMID- 2986603 TI - H+-gradient-dependent active transport of tetraethylammonium cation in apical membrane vesicles isolated from kidney epithelial cell line LLC-PK1. AB - Transport of [3H]tetraethylammonium (NEt4+), an organic cation, has been studied by using apical-membrane vesicles isolated from cultured kidney epithelial cell line LLC-PK1. The uptake of NEt4+ by apical-membrane vesicles was osmotically sensitive, time-dependent and saturable. The presence of an H+ gradient ([H+]i greater than [H+]o) induced a marked stimulation of NEt4+ uptake against its concentration gradient (overshoot phenomenon), and this concentrative uptake was inhibited by HgCl2. These results suggest that apical membranes isolated from the LLC-PK1 cells retain the transport characteristics of NEt4+ similar to those observed in renal brush-border membranes. PMID- 2986601 TI - The receptor function of the Na+, K+-activated adenosine triphosphatase system. PMID- 2986604 TI - Mechanism of the inhibition of cholesterol biosynthesis by 6-fluoromevalonate. AB - 6-Fluoromevalonate blocks the incorporation of mevalonic acid, but not that of isopentenyl pyrophosphate, into non-saponifiable lipids in a rat liver multienzyme system. With 3H-labelled 6-fluoromevalonate, it was found that 6 fluoromevalonate is converted to its phospho and pyrophospho derivatives in this system. The kinetics of the two kinases were studied. 6-Fluoromevalonate 5 pyrophosphate is a potent competitive inhibitor of pyrophosphomevalonate decarboxylase (Ki 37 nM). In the multienzyme assay for cholesterol biosynthesis, there is accumulation of mevalonate 5-phosphate and mevalonate 5-pyrophosphate in the presence of 5 microM-6-fluoromevalonate, and 6-fluoromevalonate 5 pyrophosphate is more effective than 6-fluoromevalonate in inhibiting cholesterol biosynthesis. We suggest therefore that 6-fluoromevalonate blocks cholesterol biosynthesis at the level of pyrophosphomevalonate decarboxylase after being pyrophosphorylated. PMID- 2986605 TI - The lactose/H+ carrier of Escherichia coli: lac YUN mutation decreases the rate of active transport and mimics an energy-uncoupled phenotype. AB - The Escherichia coli K12 strain X71-54 carries the lac YUN allele, coding for a lactose/H+ carrier defective in the accumulation of a number of galactosides [Wilson, Kusch & Kashket (1970) Biochem. Biophys. Res. Commun. 40, 1409-1414]. Previous studies proposed that the lower accumulation in the mutant be due to a faulty coupling of H+ and galactoside fluxes via the carrier. Immunochemical characterization of the carriers in membranes from mutant and parent strains with an antibody directed against the C-terminal decapeptide of the wild-type carrier leads to the conclusion that the mutant carrier is similar to the wild-type in terms of apparent Mr, C-terminal sequence, and level of incorporation into the membrane. The pH-dependence of galactoside transport was compared in the mutant and the parent. At pH 8.0-9.0, mutant and parent behave similarly with respect to the accumulation of beta-D-galactosyl 1-thio-beta-D-galactoside and to the ability to grow on the carrier substrate melibiose. At pH 6.0, both the maximal velocity for active transport and the level of accumulation of beta-D-galactosyl 1-thio-beta-D-galactoside are lower in the mutant. The mutant also is unable to grow on melibiose at pH 5.5. However, at pH 6.0 and low galactoside concentrations, the symport stoichiometry is 0.90 H+ per galactoside in the mutant as compared with 1.07 in the parent. These observations suggest that symport is normal in the mutant and that the lower rate of transport in the mutant is responsible for the phenotype. At higher galactoside concentrations, accumulation is determined not only thermodynamically but also kinetically, contrary to a simple interpretation of the chemiosmotic theory. Therefore lower rates of active transport can mimic the effect of uncoupling H+ and galactoside symport. Examination of countertransport in poisoned cells at pH 6.0 reveals that the rate constants for the reorientation of the loaded and unloaded carrier are altered in the mutant. The reorientation of the unloaded carrier is slower in the mutant. However, the reorientation of the galactoside-H+-carrier complex is slower for substrates like melibiose, but faster for substrates like lactose. These findings suggest that lactose-like and melibiose-like substrates interact with the carrier in slightly different ways. PMID- 2986606 TI - Pyrophosphate:fructose 6-phosphate 1-phosphotransferase and glycolysis in non photosynthetic tissues of higher plants. AB - The activity of pyrophosphate:fructose-6-phosphate 1-phosphotransferase [PFK (PPi); EC 2.7.1.90] in extracts of the storage tissues of leek (Allium porrum), beetroot (Beta vulgaris) and roots of darnel (Lolium temulentum) exceeded 0.15 mumol/min per g fresh wt. As net flux from fructose 1,6-bisphosphate to fructose 6-phosphate in these tissues is unlikely, it is suggested that PFK (PPi) does not contribute to gluconeogenesis or starch synthesis. The maximum catalytic activities of PFK (PPi) in apex, stele and cortex of the root of pea (Pisum sativum) and in the developing and the thermogenic club of the spadix of cuckoo pint (Arum maculatum) were measured and compared with those of phosphofructokinase, and to estimates of the rates of carbohydrate oxidation. PPi and fructose 2,6-bisphosphate in Arum clubs were measured. The above measurements are consistent with a glycolytic role for PFK (PPi) in tissues where there is marked biosynthesis, but not in the thermogenic club of Arum. The possibility that PFK (PPi) is a means of synthesizing pyrophosphate is discussed. PMID- 2986607 TI - A study of one of the iron-sulphur clusters in oxidized hydrogenase from Megasphaera elsdenii by magnetic-circular-dichroism spectroscopy. AB - The m.c.d. spectrum of the oxidized state of hydrogenase from Megasphaera elsdenii has been measured at liquid-helium temperatures. This oxidation state of the enzyme displays a characteristic rhombic e.p.r. signal with g-values of 2.101, 2.052 and 2.005 assigned previously to a [4Fe-4S]3+ cluster as in oxidized HiPIP (high-potential iron-sulphur protein) [Van Dijk, Grande, Mayhew & Veeger (1980) Eur. J. Biochem. 107, 251-261]. The low-temperature m.c.d. spectrum shows no features attributable to an oxidized four-iron cluster of the HiPIP type, but does reveal broad, positive peaks at 460 and 730 nm, which magnetize in a manner untypical of a spin S = 1/2 cluster with g-values close to 2. The m.c.d. spectrum is most closely similar to that of dye-oxidized P-clusters known in the enzyme nitrogenase. It is therefore proposed that the rhombic e.p.r. spectrum at a g value close to 2 arises from an m.c.d.-silent radical species that may be related chemically to the cysteine persulphide species, RS-S., recently found in the hexacyanoferrate-oxidized seven-iron ferredoxin of Azotobacter vinelandii [Morgan, Stephens, Devlin, Stout, Melis & Burgess (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 1931-1935]. PMID- 2986609 TI - Morphological investigations in human islets of Langerhans isolated by the Velcro technic. AB - The lower yield of viable, isolated islets from one donor pancreas and the immunogenicity are responsible for disappointing results in human islet transplantation. Our preliminary results in the application of Velcro-technic (Lacy, 1982) studied by morphological investigations demonstrate the feasibility of this technic in human islet isolation. If we compare the influence of Velcro technic with regard to the first step of Velcro-technic, the duct perfusion, we can conclude that A-cell granules, stained by Grimelius, are reduced during the collagenase perfusion. On the other side, we recognized B-cell granules up to 60 min after duct perfusion. Connective tissue is reduced by the isolation technic and absent completely up to the Ficoll separation. The exocrine tissue of the pancreas is damaged continuously, and after Ficoll separation we detected singular acinar cells only. The yield from the used lienal human pancreas were about 80 000 islets. Islets after Ficoll separation, studied by electron microscopic investigation, showed non-damaged A and B cells with granules in the typical manner. PMID- 2986610 TI - The role of poly(ADP-ribose) synthetase in the development of insulin-dependent diabetes and islet B-cell regeneration. PMID- 2986608 TI - Chemiluminescence in neutrophils and Lettre cells induced by myxoviruses. AB - Luminol-mediated chemiluminescence in neutrophils is stimulated by Sendai virus and by influenza virus; Lettre cells also exhibit chemiluminescence (less than 10% of that of neutrophils), which is stimulated by Sendai virus and by influenza virus. Virally induced permeability changes are not responsible for chemiluminescence, since (i) extracellular Ca2+ inhibits permeability changes but stimulates chemiluminescence, and (ii) influenza virus, which induces permeability changes at pH 5.3 but not at pH 7.4, induces chemiluminescence at either pH. Other agents [zymosan, N-formyl-L-methionyl-L-leucyl-L-phenylalanine, 4-phorbol 12-myristate 13-acetate (phorbol ester), A23187] likewise induce chemiluminescence in the absence of permeability changes. PMID- 2986611 TI - Hyperglycemia in BALB/c mice after pretreatment with one subdiabetogenic dose of streptozotocin and subsequent infection with a Coxsackie B4 strain. AB - Male BALB/c mice were injected with one subdiabetogenic dose of streptozotocin followed by Coxsackie B4 virus infection 7 days later. The animals developed a transient hyperglycemia after streptozotocin-pretreatment and infection with a human Coxsackie B4 isolate. Frozen sections of pancreata stained with FITC labeled antibodies showed an intensive infection of the exocrine tissue. Immunofluorescence studies with isolated islets obtained from streptozotocin treated or untreated animals demonstrated virus antigen in about 20% of the islets 5 days after in vivo virus infection. It is supposed that the hyperglycemia measured in our experiments was caused by a cumulative effect of streptozotocin and virus infection. PMID- 2986612 TI - Identification of an HLA-DQ beta-chain related genomic sequence associated with insulin-dependent diabetes. AB - Restriction fragment length polymorphism detected by a cDNA probe for an HLA-DQ beta-chain gene has revealed a HLA-DR4 linked BamH1 3.7 kb fragment which is rarely found among insulin-dependent diabetic patients. The present analysis demonstrates that the BamH1 3.7 kb fragment present on a HLA-DR4 positive chromosome in a healthy individual contains coding sequences for an HLA-DQ beta chain gene and that the absence of this fragment among HLA-DR4 positive insulin dependent patients is due to the loss of a BamH1 restriction within an intervening sequence. PMID- 2986613 TI - Proton microprobe analysis of pancreatic beta-cells. AB - Freeze-dried pancreas sections from obese-hyperglycemic mice were subjected to proton bombardment and the elemental contents in the beta-cells and the exocrine part were obtained from the characteristic X-rays emitted. Quantitative data were provided for 18 different elements. The mole ratio between K and Na exceeded 10, implying that neither the sample preparation nor the irradiation had induced significant diffusive changes. With the demonstration of this high K/Na ratio it seems likely that also the beta-cells are equipped with an efficient Na+/K+ pump. The beta-cells contained about 70 mmoles Cl per litre cell water. Observed amounts of Ca and Mg were equivalent to those previously recorded by electrothermal atomic absorption spectroscopy. The significant role of Zn for the storage of insulin was emphasized by the demonstration of 3 times as much of this element in the beta-cells as compared with the exocrine pancreas. In addition, the sensitivity of the proton microprobe enabled measurements of various trace elements such as Rb, Cr, Cu, Al and Pb not previously demonstrated in the pancreatic beta-cells. PMID- 2986614 TI - Role of calcium in defective insulin secretion from human and transplantable rat islet cell tumours. AB - Regulation of B-cell function was examined using pieces of a benign islet cell tumour from a 60-year-old woman, and isolated cells from a serially transplantable rat insulinoma. Acute incubations were performed after 3-days culture in RPMI-1640 containing 11.1 mM glucose. A variety of nutrients and drugs with established effects on pancreatic B-cell function failed to affect 45Ca uptake or insulin release by human tumour pieces. These included glucose alone or in combination with theophylline, glyceraldehyde, mannoheptulose, diazoxide, KCl, verapamil, D-600 and trifluoroperazine. The ineffectiveness of diazoxide was confirmed in long term cultures. Freshly isolated or 3-day cultured rat insulinoma cells also failed to respond to glucose with increased 45Ca uptake and insulin release. The results indicate an important role of calcium in the defective stimulus-secretion coupling of insulin-producing islet cell tumours. PMID- 2986615 TI - Human growth hormone binding and stimulation of insulin biosynthesis in cloned rat insulinoma cells. AB - Binding of 125I labelled human growth hormone to cloned insulin producing RIN-5AH cells is described. Binding was specific for somatotropic hormones since both human and rat growth hormone could compete for binding sites, whereas much higher concentrations of lactogenic hormones were needed to inhibit binding. Culture of RIN-5AH cells in the presence of hGH stimulated insulin biosynthesis by 85%. PMID- 2986616 TI - Role of AMP deaminase reaction in the control of fructose 1,6-bisphosphatase activity in yeast. AB - The physiological role of the inhibition of AMP deaminase (EC 3.5.4.6) by Pi was analyzed using permeabilized yeast cells. (a) Fructose 1,6-bisphosphatase (EC 3.1.3.11) was inhibited only a little by AMP, which was readily degraded by AMP deaminase under the in situ conditions. (b) The addition of Pi, which showed no direct effect on fructose 1,6-bisphosphatase, effectively enhanced the inhibition of the enzyme by AMP increased through the inhibition of AMP deaminase. (c) Pi activated phosphofructokinase (EC 2.7.1.11) and inhibited AMP deaminase activity. AMP deaminase reaction can act as a control system of fructose 1,6-bisphosphatase activity and gluconeogenesis/glycolysis reaction through the change in the AMP level. Pi may contribute to the stimulation of glycolysis through the inhibition of fructose 1,6-bisphosphatase by the increase in AMP in addition to the direct activation of phosphofructokinase. PMID- 2986617 TI - Control of respiration in sonicated cytochrome oxidase proteoliposomes by gated and ungated ionophores. AB - Respiration of cytochrome oxidase-containing sonicated proteoliposomes is partially stimulated by nigericin or by alamethicin. Valinomycin at low concentrations fully releases the nigericin-dependent respiration but inhibits the alamethicin-dependent respiration. Respiratory stimulation by the gated ionophore alamethicin must occur in the face of a continuing membrane potential; stimulation by nigericin is accompanied by an increase in the measured membrane potential. We conclude that delta pH rather than delta psi may be the main source of respiratory control in our type of proteoliposomes. PMID- 2986618 TI - Light-dependent reduction of copper(II) and its effect on cell-mediated, thiol dependent superoxide production. AB - The thiol-dependent reduction of copper(II) and associated superoxide radical production in copper(II)-treated cultures of the unicellular alga Dunaliella tertiolecta is induced by light of wavelength greater than 425 nm. We propose that the effect of light on these reactions is mediated at the copper(II) reduction step through a light-stimulated ligand to metal charge transfer reaction between sulphur and copper(II). These results suggest that light may be an important controlling factor in metal-induced lipid peroxidation reactions. PMID- 2986619 TI - Studies on four cellular proto-oncogenes and their expression in PCC4 embryonal carcinoma cells: amplification of c-Ki-ras oncogene. AB - We examined the expression of four cellular proto-oncogenes in two teratocarcinoma cell lines, one undifferentiated and malignant (PCC4), one differentiated and non malignant (PCD1). In this paper we report that transcript levels of c-Ki-ras, c-myc and c-fos are significantly higher in PCC4 cells whereas transcripts of c-Ha-ras are unchanged. Southern blot analysis does not reveal any structural alteration of c-myc and c-fos in PCC4 cells. On the contrary, the c-Ki-ras proto-oncogene is amplified 10 to 20 fold in PCC4 cells. PMID- 2986621 TI - Regulation of synthesis and activity of bovine adrenocortical NADPH-cytochrome P 450 reductase by ACTH. AB - Primary, monolayer cultures of bovine adrenocortical cells have been utilized to demonstrate that ACTH regulates the synthesis and thus the activity of NADPH cytochrome P-450 reductase. The temporal pattern of induction correlates well with that previously reported for cytochromes P-45017 alpha and P-450 C21, microsomal steroid hydroxylases serviced by P-450 reductase. Results of in vitro translation studies suggest that ACTH regulates P-450 reductase synthesis at the transcriptional level and show that the adrenocortical enzyme is synthesized as the mature form. The action of ACTH to induce P-450 reductase synthesis in the adrenal cortex can be mimicked by dibutyryl cAMP, but not by phenobarbital which induces P-450 reductase synthesis in liver. PMID- 2986620 TI - Phorbol 12-myristate, 13-acetate potentiates the action of the calcium ionophore in stimulating arachidonic acid release and production of phosphatidic acid in rabbit neutrophils. AB - The addition of the tumor-promoting phorbol 12-myristate, 13-acetate to rabbit neutrophils greatly potentiates the effect of the calcium ionophore A23187 on [3H]-arachidonic acid release and [32P]-phosphatidic acid generation. At 5 X 10( 8) M A23187, the addition of 20 ng/ml PMA potentiates the action of the ionophore on [3H]-arachidonic acid release by 5-fold. At 5 X 10(-7) M A23187, PMA enhances [32P]-phosphatidic acid production by 1.5-fold. Incubation of the neutrophils with 5 X 10(-7) M ionophore for two minutes causes a significant increase in the [32P] phosphatidic acid production but does not affect the levels of [32P] phosphatidylinositol or [32P]-phosphatidylinositol 4,5 bis-phosphate. In addition, increasing the sodium chloride concentrations in the suspending medium causes an increase in the level of phosphatidylinositol 4,5 bis-phosphate. These results suggest that the phorbol ester either acting directly or through the activation of protein kinase C modulates significantly the activities of the various forms of phospholipases, particularly A2, and/or increases the availability or amounts of their substrates. PMID- 2986622 TI - Carbachol induces desensitization of cardiac beta-adrenergic receptors through muscarinic M1 receptors. AB - Incubation of enzymatically dissociated cardiac myocytes with carbachol leads to a time- and concentration-dependent loss of beta-receptors assayable with [3H]CGP 12177. This loss is due to a redistribution of beta-receptors from the plasma membrane to a cytosol-derived vesicular fraction, consistent with an internalization process. The carbachol effects are not influenced by gallamine or oxotremorine which interact with the high-affinity (M2) muscarinic receptors. These results suggest that carbachol-induced desensitization is secondary to activation of protein kinase C by diacylglycerols generated through M1 receptor linked phosphoinositide hydrolysis. PMID- 2986623 TI - Separation of rabbit liver latent and spontaneously active phosphorylase phosphatases by chromatography on heparin-sepharose. AB - Latent and spontaneously active forms of phosphorylase phosphatase were separated by heparin-Sepharose chromatography of rabbit liver extract. The latent enzyme had an absolute polycation (histone H1, polybrene) requirement for the activity assayed with phosphorylase a and phosphorylase kinase substrates. Ethanol treatment resulted in the activation of both phosphatases by dissociating of 150 180 kDa holoenzymes to 33-38 kDa catalytic subunits as judged by gel filtration. The latent and spontaneously active phosphatases were differentiated according to their abilities to dephosphorylate the alpha and the beta subunits of phosphorylase kinase and sensitivities to inhibition by inhibitor-2 or heparin, and were classified as type-2A and type-1 phosphatases, respectively. PMID- 2986624 TI - Metabolism of leukotriene B4 in hepatic microsomes. AB - Leukotriene B4 was metabolized in rat hepatic microsomes to two products. Mass spectral analysis of these two metabolites indicated that the major metabolite was the 20-hydroxy metabolite while the minor metabolite was the 19-hydroxy metabolite. The formation of these metabolites required NADPH and was linear with time (20 min) and protein (1.6 mg/ml). The Km apparent and Vmax for omega hydroxylation of LTB4 was 14 uM and 0.138 nmol/min/mg protein. In contrast, the km and Vmax for omega minus one hydroxylation was 54 uM and 0.093 nmol/min/mg protein. These results suggest that omega and omega minus one hydroxylations of LTB4 may be mediated by different isozymes of hepatic P-450. PMID- 2986625 TI - Interaction with mitochondrial membranes of a synthetic peptide with a sequence common to extra peptides of mitochondrial precursor proteins. AB - A hepta-peptide, Arg-Leu-Leu-Pro-Ser-Leu-Gly, which has a sequence involved in the extra peptides of mitochondrial proteins, was synthesized chemically. The peptide was found to bind specifically to mitochondria, but not to microsomes. The binding was blocked by pretreatment of mitochondria with trypsin but was not affected by the presence of apocytochrome c. The synthetic peptide inhibited the binding to mitochondria of the precursor protein of ATPase inhibitor, which was synthesized in vitro, but did not inhibit that of the precursor of the 9 K stabilizing factor, which has an entirely different extra-peptide sequence. The peptide also did not inhibit the binding of apocytochrome c. These results suggest the existence of a common protein receptor on mitochondrial membranes that facilitates entrance of a group of mitochondrial precursor proteins, including pre-ATPase inhibitor. PMID- 2986626 TI - Amino acid sequence of cytochrome c-552 from Thermus thermophilus HB8. AB - The complete amino acid sequence of thermophilic cytochrome c-552 from Thermus thermophilus HB8 is presented. The 131-residue sequence was derived by analysis of three cyanogen bromide fragments of the S-carboxymethylated apo-protein and their subpeptides. The sequence is homologous to c-type cytochromes, especially in the heme-binding region. PMID- 2986627 TI - Thermodynamics of agonist and antagonist binding to the alpha 1-adrenoceptor studied using [125I]BE 2254. AB - The thermodynamics of binding of [125I]BE 2254 to the alpha 1-adrenoceptor in guinea pig brain membranes have been investigated at four different temperatures between 0 and 37 degrees C. The affinity and binding capacity of the radioligand did not vary with temperature. Thus, the change in enthalpy upon binding was close to zero whereas the change in entropy was large and positive (delta S degrees approximately 45 cal/mol-deg). In addition, [125I]BE 2254 has been used as a reporter ligand to probe the thermodynamics of the interaction of a variety of alpha-adrenoceptor agonists and antagonists with the alpha 1-adrenoceptor. Binding of all ligands was associated with large positive changes in entropy (delta S degrees between 18 and 48 cal/mol-deg) and little, or no, change in enthalpy, a finding that provides no convincing evidence for conformational rearrangement of alpha 1-adrenoceptors upon ligand binding. PMID- 2986628 TI - Detergent solubilization of human neutrophil leukotriene B4 receptors. AB - Specific leukotriene B4 (LTB4) receptors in human neutrophils were solubilized by treatment of "receptor fraction" membranes with the zwitterionic detergent (3-[(3 cholamidopropyl)-dimethylammonio]1-propane sulfonate (CHAPS). The soluble receptors were assayed by polyethylene glycol (PEG) precipitation coupled with Millipore filtration. The solubilized receptors retained all of the characteristics of the receptor sites in intact neutrophils. The binding of LTB4 was rapid, reversible and stereospecific. Mathematical modeling analysis revealed biphasic binding of [3H] LTB4 indicating two classes of binding sites. The high affinity binding site had a dissociation constant of 1.93 nM and Bmax of 281 fmoles/mg protein; the low affinity binding site had a dissociation constant of 78.92 nM and Bmax of 2522 fmoles/mg protein. Competitive binding experiments with structural analogs of LTB4 demonstrate that the interaction between LTB4 and its binding site is stereospecific and correlates with the relative biological activity of the analogs. These data suggest that it may be possible to purify the LTB4 receptor from human neutrophil membranes. PMID- 2986629 TI - MDA-468, a human breast cancer cell line with a high number of epidermal growth factor (EGF) receptors, has an amplified EGF receptor gene and is growth inhibited by EGF. AB - Epidermal growth factor (EGF) has been noted to stimulate proliferation of a variety of normal and malignant cells including those of human breast epithelium. We report here that MDA-468, a human breast cancer cell line with a very high number of EGF receptors, is growth-inhibited at EGF concentrations that stimulate most other cells. The basis for the elevated receptor level is EGF receptor gene amplification and over-expression. An MDA-468 clone selected for resistance to EGF-induced growth inhibition shows a number of receptors within the normal range. The results are discussed in relation to a threshold model for EGF-induced growth inhibition. PMID- 2986630 TI - Effect of a novel thermogenic beta-adrenoceptor agonist (BRL 26830) on insulin resistance in soleus muscle from obese Zucker rats. AB - Young lean (Fa/?) and obese (fa/fa) rats were treated with the thermogenic beta adrenoceptor agonist, BRL 26830, for 3 weeks. In lean rats this treatment had no effect on body weight but there was a marked increase in the insulin sensitivity of soleus muscle strips with respect to glycolytic rate. Treatment of obese rats with BRL 26830 produced a small but not significant decrease in body weight but the sensitivity of both glycolysis and glycogen synthesis to insulin was increased so that muscles of treated obese rats showed similar insulin sensitivity to untreated lean rats. It is suggested that such changes are unlikely to be merely a secondary consequence of an anti-obesity action. PMID- 2986631 TI - Light-mediated breakdown of phosphatidylinositol-4,5-bisphosphate in isolated rod outer segments of frog photoreceptor. AB - When isolated frog (Rana catesbeiana) rod outer segment (ROS) fragments were incubated with [gamma-32P]ATP in the dark, only two of phospholipids, i.e., phosphatidylinositol-4-phosphate (DPI) and phosphatidic acid (PA) incorporated 32P. Upon addition of DPI (100 microM), considerable amount of 32P was incorporated into phosphatidylinositol-4,5-bisphosphate (TPI) as well as DPI and PA. Exposure of the ROS membranes to 5 sec flash of light resulted in approx. 20% decrease in the labeled TPI, while no significant effect was observed on DPI and PA. It was also observed that Ca2+ markedly accelerated the production of PA in the dark, while it reduced the 32P-incorporation into TPI. These results suggest that there is light- and/or Ca2+-dependent TPI-specific phospholipase C in ROS of vertebrate photoreceptors. PMID- 2986632 TI - A metabolite of aspartame inhibits angiotensin converting enzyme. AB - Aspartame (L-aspartyl-L-phenylalanine methyl ester, is a widely used artificIal sweetener. In humans and other animals aspartame is initially hydrolyzed to L aspartyl-L-phenylalanine by intestinal esterases. L-Aspartyl-L-phenylalanine inhibits angiotensin converting enzyme purified from rabbit lungs with a Ki of 11 +/- 2 microM, equipotent to the IC50 of 12 microM for 2-D-methyl-succinyl-L proline which has been reported to be an orally active antihypertensive agent in rats. Thus the possibility exists that L-aspartyl-L-phenylalanine inhibits angiotensin converting enzyme in humans consuming large quantities of aspartame. Both aspartame itself and the diketopiperazine formed from it, 3-carboxymethyl-6 benzyl-2,5-diketopiperazine, are weak inhibitors with Ki's greater than 1 mM. PMID- 2986634 TI - Evidence for the existence of hydrogen-bonded cyclic tetramers in aqueous solutions of tetramethylammonium guanosine 5'-monophosphate. AB - Proton NMR and FTIR measurements have shown that the formation of stable, interbase hydrogen-bonded species of 5'-GMP in aqueous solutions of the tetramethylammonium salt can occur without the subsequent formation of ordered, slowly-exchanging structures or base stacking. Analysis of the NMR data is in agreement with an equilibrium mixture of monomers, dimers and cyclic tetramers. The association constants and amino proton chemical shifts are reported for these species. This is the first report that the cyclic tetramer structure exists as a discrete species in 5'-GMP solutions. PMID- 2986633 TI - Specific binding sites for platelet activating factor in human lung tissues. AB - Specific and saturable binding of [3H]-labeled 1-0-alkyl-2-0-acetyl-sn-glycero-3 phosphocholine (PAF) to membrane preparations of human lung tissues is demonstrated. The equilibrium dissociation constant (KD) was determined by Scatchard analysis to be 4.9 (+/- 1.7) X 10(-10)M and the maximal number of binding sites was estimated to be 140 (+/- 37) fmole/mg protein. The binding site is PAF specific and its selectivity toward PAF analogs is very similar to that in rabbit platelets. Two PAF receptor antagonists, kadsurenone and ginkgolide B, previously characterized in platelet systems, also displace the binding of [3H] PAF to human lung homogenates. These data indicate that human lung tissues contain PAF specific receptors, and binding of PAF to these receptor sites may be the first step to initiate PAF-induced lung pathophysiology. PMID- 2986635 TI - Evidence for heterodimeric nature of calpain molecules as studied by cross linking modification. AB - Purified calpain I and calpain II from porcine erythrocytes and kidney were cross linked with a bifunctional reagent, disuccinimidyl suberate, and the cross-linked products were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The major product had a molecular mass of 105 kDa, while the starting materials were resolved into 80-kDa and 30-kDa subunits. The cross linking in the presence of 2 mM Ca2+ yielded several higher-molecular-weight species. The cross-linked products were shown to contain both the 80-kDa and 30 kDa proteins by means of immunoblotting with antibodies monospecific for the respective subunits, suggesting that the original calpain molecule existed in solution as an 80-kDa plus 30-kDa heterodimer and that Ca2+ induced closer association of these heterodimeric molecules. PMID- 2986637 TI - Differentiation and de-differentiation of cultured chondrocytes: increase in monomeric size of 'cartilage-specific' proteoglycans by dibutyryl cyclic AMP and complete inhibition of their synthesis by retinoic acid. AB - Dibutyryl cyclic AMP (DBcAMP) induced an increase in the monomeric size of 'cartilage-specific' proteoglycans (PG-I) in rabbit costal chondrocytes in culture. This increase in size was due to an increase in the average molecular weight of the glycosaminoglycan (GAG) chains. In contrast, retinoic acid completely inhibited the synthesis of PG-I. However, the synthesis and monomeric size of 'ubiquitous' proteoglycans (PG-II) were little affected by these agents. These results suggested that modulation of the differentiated state of chondrocytes is closely related to not only the synthesis of 'cartilage-specific' proteoglycans but also their monomeric size. PMID- 2986636 TI - The effect of adenylate kinase activity on the rate and efficiency of energy transport from mitochondria to hexokinase. AB - The rate and efficiency of energy transport were examined in a system containing isolated rabbit heart mitochondria, hexokinase, adenylate kinase and low concentrations of adenine nucleotides. Oxygen consumption by mitochondria and glucose-6-phosphate synthesis by hexokinase were registered. It was found that when adenylate kinase is active both in mitochondria and in the environmental solution, the rate and efficiency (glucose-6-phosphate/O ratio) of glucose-6 phosphate formation considerably increase. The effects of adenylate kinase activity are fully abolished by diadenosine pentaphosphate, an inhibitor of adenylate kinase. PMID- 2986638 TI - DNA strand scission by enzymatically reduced mitomycin C: evidence for participation of the hydroxyl radical in the DNA damage. AB - Phage DNA, as well as plasmid and mammalian DNA's, were exposed to a superoxide and hydroxyl radical-generating system containing NADPH-cytochrome P-450 reductase and mitomycin C, both with and without added Fe3+-ADP, in phosphate buffer at pH 7.5. The generation of superoxide (O2-.) and hydroxyl (.OH) radicals in the system was demonstrated by using ESR spectrometry with N-tert -butyl-alpha phenylnitrone (PBN) as a spin trapping agent. Only the lambda DNA isolated after exposure to the O2-./.OH-generating system containing many lower molecular weight DNA fragments indicating DNA strand breaks. This breakage was completely inhibited by a .OH radical scavenger (sodium benzoate) and by catalase, but only slightly by superoxide dismutase. Thyroid and plasmid DNA's were both cleaved when exposed to the O2-./.OH-generating systems. It is suggested that the mechanism of DNA scission by mitomycin C described here closely resembles that induced by the anthracycline drugs. PMID- 2986639 TI - A novel approach for the study of thromboxane A2 and prostaglandin H2 receptors using an 125I-labeled ligand. AB - Previous studies of eicosanoid receptors have utilized 3H-labeled ligands. However, 125I has a higher theoretical specific activity (approximately 2000 Ci/mmole) than 3H (29 Ci/mmole), which provides a potential advantage for 125I ligand binding studies when the receptor density is low. Since eicosanoids do not possess an easily iodinatable structure (e.g. a phenol or imidazole ring), it is not feasible to directly incorporate 125I into the molecule. The thromboxane A2/prostaglandin H2 receptor antagonist, cis-7-(2-p hydroxyphenylethanolaminocyclopentyl)-heptanoic acid (cis-APO), was synthesized to test the concept that it could be labeled with 125I and used as a ligand for binding studies. cis-APO is a structural analog of 13-azaprostanoic acid, a TXA2/PGH2 antagonist [G. C. Le Breton, D. L. Venton, S. E. Enke and P. V. Halushka, Proc. natn. Acad. Sci. U.S.A. 76, 4097 (1979)], in which the omega aliphatic chain was substituted with 2-p-hydroxyphenylethanol, which contains a phenolic group. [127I]cis-APO was synthesized by insertion of 127I (stable isotope) into the phenolic portion of the molecule under alkaline conditions. [125I]-cis-APO was synthesized via insertion of 125I (unstable isotope) into the molecule in the presence of chloramine T. cis-APO inhibited human platelet aggregation induced by the thromboxane mimetic U46619 [C. Malmsten, Life Sci. 18, 169 (1976)]. The IC50 for cis-APO was 6.4 +/- 0.7 microM and for [127I]-cis-APO was 9.8 +/- 1.3 microM (P less than 0.001). [125I]-cis-APO binding to a human platelet membrane preparation at 4 degrees was time and protein concentration dependent, saturable, and reduced or abolished by trypsin or boiling respectively. The Kd for iodo-cis-APO determined at equilibrium using a Scatchard analysis was 1.48 microM and the maximum binding capacity was 18.7 pmoles/mg protein. The forward rate constant (k+1) was 2.3 X 10(3) M-1 s-1 and the dissociation constant (k-1) was 2.12 X 10(-3) s-1. The Kd determined from k-1/k+1 was 0.92 microM. These observations show that the omega side chain of eicosanoid analogs can be substituted with a phenolic group, iodinated, and retain biological activity. These molecules may then be utilized to study thromboxane A2 or prostaglandin H2 receptors. PMID- 2986640 TI - Effect of an adrenocorticotropin analogue, ACTH 1-17, on DNA synthesis in murine metaphyseal bone. AB - The effects of injections of a synthetic adrenocorticotropin (ACTH 1-17, Synchrodyn) on the rate of DNA labeling in the metaphyseal bone of CD2F1 mice were tested on a chronopharmacological dosing schedule. Groups of mice that had been conditioned to a 12-hr light/12-hr dark schedule were injected at one of six different timepoints, 4 hr apart, during a single 24-hr span with either a low (0.02 I.U./kg) or a high (20 I.U./kg) dose of ACTH 1-17. Control groups received injections of a placebo at corresponding timepoints. Subgroups of mice were injected with [3H]thymidine ([3H]Tdr) to follow the changes in DNA labeling in the proximal tibial metaphysis at 15 min and 2, 4, 8, 12 and 24 hr after ACTH 1 17 or placebo treatment. All mice were injected with the isotope 30 min before killing, except for those killed 15 min after Rx administration where the isotope had been injected 14 min before killing. The data were analyzed both by analysis of variance and by the cosinor method, the latter of which tests the fit of a 24 hr cosine curve to the data. The effect of ACTH 1-17 on the target cell population was dependent not only upon the dose but upon the time of administration. Both doses exerted time-dependent action, ranging from stimulation to inhibition of DNA labeling. Inhibition was noted when the ACTH 1 17 was administered at 2 hr after the beginning of the daily dark span when nocturnal animals become active. When administered at this circadian stage, the larger dose in particular was associated with an inhibition of DNA labeling lasting for 24 hr. The inhibitory effect was much shorter when the same dose was injected 4 hr earlier. Moreover, the large ACTH 1-17 dose had a stimulatory effect lasting for 24 hr when it was administered 2 hr after the onset of the daily light span, with a much shorter stimulation following administration of the large dose at 6 hr after the beginning of the daily dark span. A circadian stage dependent stimulation or inhibition of DNA labeling at 2 or 14 hr after light onset, respectively, was thus complemented by an initial inhibition followed by stimulation and vice versa at 10 and 18 hr after light onset respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986641 TI - Metabolism of the amino acid beta-pyrazol-1-ylalanine and its parent base pyrazole. AB - beta-Pyrazol-1-yl-DL-alanine, an uncommon amino acid from plants of the Cucurbitaceae, was fed to mice. Although pyrazole is known to affect the liver enzymes UDP-glucose dehydrogenase, UDP-glucuronyl transferase and UDP-glucuronic acid pyrophosphatase, and also depresses their liver glycogen concentrations, beta-pyrazol-1-ylalanine had no such effects. beta-Pyrazol-1-ylalanine could not be detected in the liver of the experimental animals but was present in the urine. No other change in urinary amino acid content was observed. Studies with [14C]-beta-pyrazol-1-yl-DL-alanine showed the administered amino acid was excreted over a 4-day period, 93% of the compound supplied was recovered. Similar recoveries were obtained with the L-enantiomer from cucumber seed. The metabolic inertness of beta-pyrazol-1-ylalanine was also apparent in experiments involving subcutaneous injection of this compound. Administration of pyrazole confirmed an earlier report of resultant increased activity of liver UDP-glucose dehydrogenase and UDP-glucuronyl transferase, and of the depression of activity of liver UDP glucuronic acid pyrophosphatase. A concomitant 40% decrease in liver glycogen content was seen. The urine contained a novel metabolite, identified as a peptide conjugate of a pyrazole derivative. Mass spectrometry and p.m.r. spectroscopy indicate that this derivative is 3,4,4-trimethyl-5-pyrazolone. The amino acid constituents are aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine and leucine. The urine of mice receiving pyrazole contained less free glycine and alanine than controls. From the results, it is concluded that pyrazole is not a catabolite of dietary beta-pyrazol-1-ylalanine but to the contrary, the amino acid is essentially excreted unchanged. Formation of 3,4,4-trimethyl-5-pyrazolone from pyrazole would imply C-methylation, a process that has not been previously observed in a mammalian detoxication context. PMID- 2986642 TI - Stimulation of phosphoinositide hydrolysis and inhibition of cyclic AMP formation by muscarinic agonists in developing chick heart. AB - As early as 4 days in ovo, phosphoinositide hydrolysis in embryonic chick heart was stimulated by the cholinergic agonist carbachol. In the 4-day chick heart, the concentrations of carbachol giving half-maximal and maximal stimulation of [3H]inositol 1-phosphate formation were 30 microM and 1 mM respectively. These values are identical to those obtained using hearts from 13-day embryos [J. H. Brown and S. L. Brown, J. biol. Chem. 259, 3777 (1984)]. The amounts of [3H]inositol 1-phosphate formed per mg protein in the presence of carbachol were greater at early ages than at later ones; the stimulation by carbachol thus decreased from 8-fold at 4 days to 2-fold at 13 days. Muscarinic receptor stimulation also led to inhibition of isoproterenol-stimulated cyclic AMP formation as early as 4 days. The isoproterenol-stimulated increase in cyclic AMP was greatest at early ages, whereas the inhibitory effect of carbachol remained constant at 75%. At 4 days, half-maximal and maximal inhibition by carbachol occurred at 0.3 and 30 microM, respectively, the same values obtained using 13 day hearts (see reference cited above). Thus, the two biochemical responses of embryonic chick heart to muscarinic agonists have identical agonist sensitivities regardless of embryonic age and are functional prior to parasympathetic innervation and physiological responsiveness. PMID- 2986643 TI - Involvement of leukocyte peroxidases in the metabolism of tenoxicam. PMID- 2986644 TI - Are there neuropeptide-specific peptidases? PMID- 2986645 TI - Negative regulation of cyclic-AMP levels by carbamylcholine in dog thyroid is not mediated by cyclic-GMP. AB - Carbamylcholine, through calcium, enhances cyclic-GMP accumulation and depresses cyclic-AMP accumulation in TSH stimulated dog thyroid. The results presented show that compounds which can be transformed to nitric oxide increase cyclic-GMP accumulation in the dog thyroid. These compounds do not require extracellular calcium for their action. In thyroid stimulated by TSH, these compounds do not depress AMP accumulation. Cyclic-GMP is not the main intracellular signal involved in the negative regulation of cyclic-AMP levels in dog thyroid. PMID- 2986646 TI - Adenosine-homocysteine interactions in controlling cyclic AMP in rat brain cortical slices. AB - Preincubation of rat brain cortical slices with homocysteine resulted in a reduction of the cyclic AMP response to subsequently added noradrenaline or isoproterenol. There was no effect of homocysteine on the adenosine response, nor of added S-adenosyl homocysteine on any of the responses studied. The results are interpreted in terms of binding of endogenous adenosine by the enzyme S adenosylhomocysteinase, and provide further evidence for the controlling role of adenosine on cerebral cyclic AMP production. PMID- 2986647 TI - Comparison of the effects of some compounds on human neutrophil degranulation and leukotriene B4 and thromboxane B2 synthesis. AB - The effects of aspirin, indomethacin, phenylbutazone, eicosatetraynoic acid (ETYA), nordihydroguaiaretic acid (NDGA) and 3-amino-1-[3-(trifluoromethyl) phenyl]-2-pyrazoline (BW755C) on human neutrophil degranulation induced by A23187 and F-Met-Leu-Phe (FMLP) have been studied. These effects have been compared with those on A23187 induced leukotriene B4 (LTB4) and thromboxane B2 (TXB2) synthesis by these cells to elucidate the relationship between LTB4 formation and degranulation. All compounds inhibited TXB2 synthesis by 50% at concentrations between 0.0016 and 50 microM. The synthesis of LTB4 was inhibited by 50% by ETYA (1.9 microM) and by NDGA (0.52 microM). Degranulation induced by A23187 and FMLP was inhibited by 50% by ETYA (16 and 11 microM respectively) and by NDGA (1.5 and 6.5 microM respectively). In the case of ETYA the concentrations required to inhibit degranulation were significantly higher than those required to inhibit LTB4 synthesis. In contrast, BW755C inhibited LTB4 synthesis by 50% at 2.8 microM but did not affect A23187-induced degranulation and was only a weak inhibitor of FMLP-induced degranulation (50% inhibition at 89 microM). The effects of the above compounds on the omega-oxidation of LTB4 by human neutrophils has also been studied to investigate the mechanism of action of these compounds. None of the above compounds affected the metabolism of LTB4 by these cells suggesting that their actions are not as non-specific anti-oxidants. These data indicate that human neutrophil degranulation induced by FMLP and A23187 is independent of LTB4 synthesis. PMID- 2986648 TI - Inhibition by CV-3988 of the binding of [3H]-platelet activating factor (PAF) to the platelet. AB - The inhibitory effects of CV-3988, a specific antagonist of PAF, on the binding of [3H]-PAF to washed platelets of various species including human were examined. The dissociation constant (Kd), binding capacity (Bmax), and the number of receptor/platelet for the specific binding site of rabbit platelets were 2.2 +/- 0.2 nM, 93.7 +/- 8.3 fmoles/10(8) platelets, and 568 +/- 50, respectively. CV 3988 selectively inhibited the specific binding of [3H]-PAF to rabbit platelets with an IC50 of 7.9 X 10(-8) M, and it slightly increased the Kd value (2.5 +/- 0.8 nM) and decreased the binding capacity for PAF (Bmax: 54.3 +/- 16.3 fmoles/10(8) platelets). The Ki value of CV-3988 for the specific binding of [3H] PAF to rabbit platelets was 1.2 X 10(-7) M. CV-3988 had no effects on the binding of [3H]-5-hydroxytryptamine (5-HT) to rabbit platelets and on the shape change of the platelet induced by 5-HT. CV-3988 also inhibited the specific binding of [3H] PAF to human and guinea-pig platelets with IC50 values of 1.6 X 10(-7) and 1.8 X 10(-7) M, respectively. CV-3988 inhibited the PAF-induced aggregation in rabbit, guinea-pig, and human platelets. These findings show that CV-3988 is a specific antagonist of PAF at the receptor site(s) of platelets and, in these species, inhibits PAF-induced platelet aggregation by inhibiting the binding of PAF to the "PAF receptor". No specific binding of [3H]-PAF to the platelet of rats and mice was observed, indicating that these species lack a PAF receptor. PMID- 2986649 TI - Investigations of the action of the antitumour drug adriamycin on tumour cell membrane functions--I. AB - The membrane potential of L1210 murine leukemia cells was assessed by use of the tritiated lipophilic cation probe triphenylmethylphosphonium bromide. The potassium equilibrium potential of the cells was found to be -71 +/- 7 mV. The resting membrane potential was partly dissipated by the protonophore m chlorocarbonylcyanidephenylhydrazone (10 microM), but was unaffected by ouabain (1 mM) and apparently by the calcium ionophore A23187 (2.5 microM). Monensin (20 microM) caused a hyperpolarization which, since it was blocked by ouabain, was presumed to be brought about by activation of the Na+K+-ATPase via an elevated cytoplasmic Na+ concentration. Adriamycin at concentrations as high as 5 X 10(-4) M brought about no change in the resting potential of the cells. Also, cytotoxic concentrations of adriamycin, unlike ouabain, had no effect on rubidium-86 transport into L1210 cells, nor upon a monensin-induced increased in rubidium-86 uptake. The results suggest that although adriamycin is capable of interaction with the plasma membrane, and may exert its cytotoxicity at this locus, changes in ion flux mediated by Na+K+-ATPase or those capable of changing the membrane potential do not appear to be implicated in its mechanism of action. PMID- 2986650 TI - Carbon tetrachloride-induced eicosanoid synthesis and enzyme release from rat peritoneal leucocytes. AB - When rat peritoneal leucocytes were incubated with carbon tetrachloride, a PLA2 was activated, eicosanoids were generated and lysosomal and cytoplasmic enzymes were released. The predominant eicosanoid generated was TXB2 with lesser amounts of PGE2, 6-keto PGF1 alpha and LTB4. Preincubation of the cells with two structurally unrelated thromboxane synthetase inhibitors reduced PLA2 activity and enzyme release and also reduced the total amounts of eicosanoids liberated. An anti-PGI2 antibody partially reversed the effects of thromboxane synthetase inhibitors indicating a role for endogenous PGI2 generation in the cytoprotective effects of these agents in this system. Exogenous PGI2 was also cytoprotective but the timing of its administration was critical. The cytoprotective effect of PGI2 was potentiated by a phosphodiesterase inhibitor, indicating a possible pivotal role of cAMP in cell protection. PMID- 2986651 TI - Increase in serotonin2 receptor density in rat cerebral cortex slices by stimulation of beta-adrenergic receptors. PMID- 2986652 TI - [The nature of cation-binding groups in binding sites for [[3H]Tyr1, D-Ala2, D Leu5]enkephalin]. AB - The influence of pH on binding of labeled stable analog of enkephalin, [3H]Tyr1, D-Ala2, D-Leu5]enkephalin, to high- and low-affinity receptors of rat brain membranes was studied. It was shown that alkali-earth metal ions combine with a deprotonated group (pKa 7,0) of the high-affinity receptor, thereby activating the latter. The effect of cations on the low-affinity enkephalin binding is independent of pH. The presence of phosphate group in the high-affinity binding site, as well as of imidazole residue in the low-affinity binding site was surmised. The latter supposition was supported by data on chemical modification of the membrane preparation with the aid of diethylpyrocarbonate. PMID- 2986653 TI - [The polypeptide composition of cytochrome oxidase from the bovine heart muscle]. AB - The beef heart cytochrome oxidase (EC 1.9.3.1) has been purified by hydrophobic chromatography. The enzyme has been resolved into 11 different polypeptides by SDS/urea gel-electrophoresis. PMID- 2986654 TI - [Effect of Bryonia cucurbitacins on the biosynthesis of eicosanoids in human leukocytes]. AB - 2 beta,25-di (beta-D-glucopyranosyl)-16 alpha,20-dihydroxy-3,11,22- trioxocucurbit-5-en and 2 beta-(beta-D-glucopyranosyl)-16 alpha,20,25-trihydroxy 3,11,22-trioxocucurbit-5-en isolated from bryonia (Bryonia alba L.) roots have been demonstrated to inhibit in vitro the [1-14C]arachidonic acid release from neutrophils. Aglicon 2 beta,16 alpha,20,25-tetrahydroxy-3,11,22-trioxocucurbit-5 en is much less active. When the cells are stimulated by calcium ionophore A23187, the aglycon potentiates the release of arachidonic acid. In these conditions the glucosides show little activity. Both the glucosides and their aglycon suppress the biosynthesis of 5S,12R-dihydroxy-6,8,10,14(Z, E, E, Z) eicosatetraenoic acid (LTB4) and 5S,12S-dihydroxy-6,8,10,14(E, Z, E, Z) eicosatetraenoic acid (5S,12S-DHETE). Inhibition of the biosynthesis of these compounds by 2 beta,16 alpha,20,25-tetrahydroxy-3,11,22-trioxocucurbit-5-en also takes place on incubation of human neutrophils with exogenous arachidonic acid. The formation of other products of cycloxygenase and lipoxygenase oxidation pathways remains practically unchanged. PMID- 2986655 TI - [In vitro activity of seven gyrase inhibitors of a group of heterocyclic carbonic acids against nonfermenting gram negative rods (nonfermenters)]. AB - MIC (minimal inhibitory concentration) determinations for nalidixic acid, cinoxacin, pipemidic acid, norfloxacin, enoxacin, and ciprofloxacin were done by agar dilution on isosensitest agar (oxoid). Bacterial strains investigated were 189 Pseudomonas aeruginosa, 164 Acinetobacter lwoffii, 4 Ps. maltophilia, 3 Ps. putrefaciens and 3 Ps. odorans. The results in summary are: Ciprofloxacin is the most active gyrase inhibitor against Ps. aeruginosa as well as against other nonfermentative gram-negative rods, versus Ps. aeruginosa norfloxacin is a little more active than ofloxacin, against nonfermentative gram-negative rods other than Ps. aeruginosa norfloxacin is markedly less active than ofloxacin. Problems concerning cross-resistance of new gyrase inhibitors are discussed. PMID- 2986656 TI - The effect of induced atherosclerosis on the synthesis of elastin in chick aortic tissue. AB - Atherosclerosis was induced in growing chickens by the administration of a diet containing elevated levels of cholesterol and vitamin D3. The effect of this diet on the accumulation of insoluble elastin and the synthesis of soluble and insoluble elastin in the thoracic aortas of these animals was measured. Although the diet resulted in significant increases in levels of cholesterol, 25-OH vitamin D3 and calcium in plasma, increased levels of cholesterol and calcium in aortic tissue, and histological evidence of aortic lipid deposition, there were no detectable differences between experimental and control animals in either the rate or the time course of accumulation of total insoluble elastin in the thoracic aorta, or in the rate and time course of synthesis of soluble and insoluble elastin. These data suggest that, at least in this model, any effect of atherosclerosis on aortic elastin production must be either small or so localized as to be not measurable by the methods used. PMID- 2986657 TI - Joseph S. Redding's contributions to cardiac resuscitation. PMID- 2986658 TI - [Etiopathogenesis of infectious diarrheas]. PMID- 2986659 TI - [Effect of methadone administration on MHV-3 virus-induced experimental hepatitis in the mouse]. AB - Mice treated with 15 mg/Kg/day methadone and infected with MHV-3 virus after 7 days did not show increased susceptibility to MHV-3 virus infection, did not develop more serious forms of hepatitis and not mortality did not increase with respect to the controls. Drug administration was continued for the duration of the experiment. PMID- 2986660 TI - The effect of calcium and 3',5' cAMP on the induction of ornithine decarboxylase activity in bone and bone cells. AB - The interrelationship between mediators of hormone action (cAMP and calcium) and induction of ornithine decarboxylase (ODC) activity was investigated in bone and bone cells. Stimuli that enhanced the cAMP level in both osteoblastlike (BL) and osteoclastlike (CL) cells and in intact calvaria only induced ODC activity in the isolated cell populations. Moreover, addition of fresh medium induced ODC activity in BL cells and not in calvaria, without an effect on the cAMP level. The tumor-promoting agent 12-O-tetradecanoyl-phorbol-13-acetate (TPA), however, induced in both systems ODC activity, without an effect on the cAMP concentration. From these data it was concluded that induction of ODC activity may not be mediated by cAMP. The role of Ca in ODC induction was also investigated. Experiments with BL cells, incubated in media with various Ca concentrations, as well as experiments with the Ca blocker D600 showed that basal and hormone-induced ODC activity was dependent on the intracellular Ca concentration. A regulatory role of cAMP, however, in concert with Ca, cannot be excluded. PMID- 2986661 TI - Oral pigmentation, Addison's disease and the results of screening for adrenocortical insufficiency. PMID- 2986662 TI - Effect of propofol, thiopentone and etomidate on adrenal steroidogenesis in vitro. AB - The i.v. anaesthetic agents propofol, thiopentone and etomidate inhibited ACTH stimulated production of cortisol by guineapig dispersed adrenal cells in a dose related manner. For two of the drugs, propofol and thiopentone, inhibition occurred over a similar concentration range: 2 X 10(-5) - 5 X 10(-4) mol litre-1. With etomidate, inhibition occurred over a much lower concentration range (5 X 10(-8) - 5 X 10(-6) mol litre-1). The concentrations of anaesthetic which induced 50% inhibition of cortisol secretion were propofol 1.7 X 10(-4), thiopentone 1.6 X 10(-4), and etomidate 1.0 X 10(-7) mol litre-1. PMID- 2986663 TI - Comparison of the effects of etomidate, thiopentone and propofol on cortisol synthesis. AB - The inhibitory effects of etomidate, thiopentone and propofol on basal and ACTH stimulated cortisol synthesis by isolated bovine adrenocortical cells have been examined. Concentrations required for 50% inhibition of ACTH-stimulated cortisol output were 1.1 X 10(-4) mol litre-1 for propofol, 3 X 10(-5) mol litre-1 for thiopentone and 4.6 X 10(-1) mol litre-1 for etomidate. At concentrations likely to be achieved during anaesthesia, etomidate would block cortisol output by isolated cells, thiopentone might slightly reduce output and propofol would be unlikely to alter cortisol synthesis significantly. PMID- 2986664 TI - Serum marker potential of placental alkaline phosphatase-like activity in testicular germ cell tumours evaluated by H17E2 monoclonal antibody assay. AB - A monoclonal antibody (H17E2) was used in a solid-phase localisation of enzyme activity (ILEA) assay to evaluate placental-like alkaline phosphatase (PLAP) as a serum marker of testicular germ cell tumours. Single or repeated assays were performed on 213 normal blood donor and a smaller number of term pregnancy and testicular cancer sera. The detection limit of PLAP by this system was 0.14 O.D. units equivalent to 0.04iul-1. Of 50 patients with established metastatic disease tested before treatment, 88% of 16 with seminoma, 54% of 13 with mixed seminoma and malignant teratoma and 33% of 21 with malignant teratoma had serum PLAP greater than 0.2 O.D. units. This compared to an incidence of 2% in non-smokers and of 29% in smokers who had been free of disease for more than 12 months. In 15 of 22 successfully treated patients, pre-treatment serum PLAP exceeded 0.2 O.D. units (mean 0.69 O.D.) and varying (53-97%) reductions in the initial levels occurred with treatment. These results with monoclonal antibody ILEA assay suggest that measurement of PLAP levels will be useful in the management of patients with germ cell tumours, particularly seminoma. PMID- 2986665 TI - Neural markers in carcinoma of the lung. AB - Small cell carcinoma (SCC) is considered to be of neuroendocrine origin. Neurone specific enolase (NSE) and PGP 9.5 are markers of neural and neuroendocrine differentiation. S-100 protein is a marker of glial differentiation. The expression of these markers in endobronchial biopsy and lung tumour resection specimens was studied to see if any diagnostic, prognostic or therapeutic implications would emerge. Zamboni fixed endobronchial tumour biopsy specimens from 20 patients were examined. Twelve of these were cases of SCC and 8 were non SCC. Of the 12 SCC, 7 were positive for NSE, 6 for PGP 9.5 and 5 for S-100 protein. Cases which showed a positive reaction for NSE had a mean survival of 9.1 months compared with 3.9 months for those with a negative reaction, but the number of cases is too small to assign any statistical significance. There was no difference in survival times between positive and negative reactors for PGP 9.5 and S-100 protein. All 8 cases of non-SCC showed positive reactions to all three markers. Of 32 formalin fixed lung tumour resection specimens 6 were cases of SCC, 25 non-SCC and 1 a chemodectoma. Three of the 6 cases of SCC showed positive staining for NSE, 3 for PGP 9.5 and 1 for S-100 protein. Of the 25 non-SCC, 10 were positive for NSE, 12 for PGP 9.5 and 6 for S-100 protein. The 1 chemodectoma stained positively for all three markers. Neuroendocrine markers are of little value in differentiating SCC from non-SCC. Positive staining for NSE in SCC may be an indicator of prolonged survival but further investigation is required. PMID- 2986666 TI - Prostacyclin and thromboxane in breast cancer: relationship between steroid receptor status and medroxyprogesterone acetate. AB - To study the production and significance of prostacyclin (PGI2) and thromboxane A2 (TxA2) in breast cancer, tissue fragments of breast cancer (n=23) and mastopathy (n=10) were superfused in vitro and the release of 6-keto-PGF1 alpha (a metabolite of PG12) and TxB2 (a metabolite of TxA2) measured by radioimmunoassay. Breast cancer formed more 6-keto-PGF1 alpha (4.5 +/- 0.9 ng min 1 g-1 of tissue dry weight, mean +/- s.e.) and TxB2 (2.5 +/- 0.6 ng min-1 g-1) (P less than 0.01) than did mastopathic breast (1.4 +/- 0.5 and 0.4 +/- 0.1 ng min-1 g-1, respectively). These productions were similar in steroid receptor positive and negative tumours. Breast cancer metastasized in 15 patients during the follow up time of 3.7 +/- 0.7 years, but the initial prostanoid productions in these patients were not different from those in nonmetastatic patients. Two patients died from metastases, but their initial mammary production of prostanoids was not profoundly different from those in the survivors. In 8 patients (4 with steroid receptor positive and 4 with negative tumour), the cancer tissue was superfused in the presence or absence of medroxyprogesterone acetate (100-5000 ng ml-1), which is commonly used for treatment of breast cancer. This hormone had no effect on mammary PGI2 and TxA2 production. We thus conclude that the PGI2 and TxA2 productions are increased in mammary cancer but that this may not be of primary significance for metastastic spread. PMID- 2986667 TI - Hepatocellular carcinoma and dietary aflatoxin in Mozambique and Transkei. AB - Estimations of the incidence of hepatocellular carcinoma (HCC) for the period 1968-74 in the Province of Inhambane, Mozambique, have been calculated and together with rates observed in South Africa among mineworkers from the same Province indicate very high levels of incidence in certain districts of Inhambane. Exceptionally high incidence levels in adolescents and young adults are not sustained at older ages and suggest the existence of a subgroup of highly susceptible individuals. A sharp decline in incidence occurred during the period of study. Concurrently with the studies of incidence, 2183 samples of prepared food were randomly collected from 6 districts of Inhambane as well as from Manhica-Magude, a region of lower HCC incidence to the south. A further 623 samples were taken during 1976-77 in Transkei, much further south, where an even lower incidence had been recorded. The mean aflatoxin dietary intake values for the regions studied were significantly related to HCC rates. Furthermore, data on aflatoxin B1 contamination of prepared food from 5 different countries showed overall a highly significant relationship with crude HCC rates. In view of the evidence that chronic hepatitis B virus (HBV) infection may be a prerequisite for the development of virtually all cases of HCC and given the merely moderate prevalence of carrier status that has been observed in some high incidence regions, it is likely that an interaction between HBV and aflatoxin is responsible for the exceptionally high rates evident in parts of Africa and Asia. Various indications from Mozambique suggest that aflatoxin may have a late stage effect on the development of HCC. This points to avenues for intervention that could be more rapidly implemented than with vaccination alone. PMID- 2986668 TI - Erionite exposure and mesotheliomas in rats. AB - Epidemiological and environmental surveys in the Cappadocian region of Turkey have linked the high incidence of pleural and peritoneal mesothelioma in the occupants of some villages with the zeolite fibres released from the locally occurring volcanic tuff. In view of the low ambient fibre concentrations and the extraordinary incidence of mesothelioma a study to test the hypothesis of high biological activity for the zeolite fibres was required. Experimental studies using both intrapleural inoculation and inhalation techniques have been undertaken with the erionite from this region and from Oregon in the United States. Additionally a non-fibrous zeolite from Japan and a synthetic non-fibrous zeolite of similar chemical composition to erionite have been included in the experiments. In these studies the samples from Oregon and Turkey produced a very high incidence of tumours. All the rats inoculated intrapleurally with Oregon erionite and almost all those inoculated with the Turkish fibre died with a mesothelioma. Inhalation of the Oregon erionite induced a similar effect. No other dusts we have investigated have produced this high incidence of tumours particularly following inhalation. These studies demonstrate that we now have a valuable new fibre for experimental study and a possible hazard to man in regions other then Turkey. PMID- 2986669 TI - The 'metabolic burst' in polymorphonuclear leukocytes from patients with quiescent psoriasis. AB - We report an investigation of peripheral blood polymorphonuclear leukocytes (PMN) from untreated patients with mild, quiescent psoriasis. Four aspects of the 'metabolic burst' were measured to illustrate phagocytosis-related events. These were: myeloperoxidase activity, hydrogen peroxide release, superoxide production and luminol-amplified chemiluminescence. Our study does not support the concept of an intrinsic abnormality of the PMN in psoriasis, but provides additional evidence for disease activity-dependent changes in phagocytic behaviour. Possible interactions with certain humoral factors, the so-called opsonins, are discussed. PMID- 2986670 TI - Development of polyneuropathy during thalidomide therapy. AB - Seven patients with prurigo nodularis and one with aphthous stomatitis were given 40-115 g of thalidomide for 1 to 6 years. They all developed a predominantly sensory peripheral neuropathy mainly involving the lower limbs. Five patients had an unpleasant tight feeling around the feet. Nerve conduction studies showed small sensory action potentials from the lower limbs with normal or only mild slowing of sensory conduction velocity indicating an axonal neuropathy. The dermatological disorder improved dramatically in all, but treatment had to be discontinued because of the severe side-effects. Thalidomide, if used, should be given only over a short period because of its neurotoxic effect. PMID- 2986671 TI - Chronic polyhydramnios is a syndrome with a lactogen receptor defect in the chorion laeve. AB - Human chorion laeve contains a lactogenic hormone [prolactin (PRL), growth hormone (GH) and placental lactogen (hPL)] receptor. Here, we studied binding of the potent lactogen, human GH, to this receptor in 18 normal pregnant women, in 12 patients whose pregnancies were complicated by chronic polyhydramnios and in 13 with chronic oligohydramnios. Polyhydramnios was classified clinically as idiopathic in seven patients, and secondary and associated with various disorders in the remaining five patients. Lactogenic hormone binding was lower in association with polyhydramnios (mean 1.60, SEM 0.15%) than with normal amniotic fluid volume (mean 3.05, SEM 0.40%; P less than 0.05); Scatchard analysis indicated that a reduced number of lactogen receptors within the chorion laeve was the reason. The lactogenic hormone receptor defect in the chorion laeve was present in pregnancies complicated by either idiopathic or secondary polyhydramnios. In contrast to chronic polyhydramnios, tissue from patients with chronic oligohydramnios bound lactogenic ligands in a normal fashion. Insulin binding sites were also identified in the chorion laeve, and, in contrast to the lactogens, binding parameters were equivalent in the three patient groups. PRL resistance developing subsequent to this chorion laeve receptor defect might produce the excessive amniotic fluid volume characteristic of polyhydramnios. We propose that a chorionic PRL receptor deficiency underlies the various clinical forms of chronic polyhydramnios. PMID- 2986672 TI - Assembly and characterization of nucleosomal cores on B- vs. Z-form DNA. AB - The ability of right- vs. left-handed alternating purine/pyrimidine copolymers to support the formation of nucleosomes has been examined by using a trout testis assembly factor. The protein, which is thermostable, has a molecular weight of 29000 and will assemble nucleosomes onto both SV40 and calf thymus DNA. This assembly factor has been used to assemble nucleosomes onto the B and Z conformations of poly[d(Gm5C)] and the B conformation of poly[d(GC)]. The isolated B-form particles, which sediment at approximately 11 S in a sucrose density gradient, contain DNA of 140-200 bases in length and the four core histones. The isolated Z-form particles, which also sediment at approximately 11 S, contain the four core histones and DNA of 170-250 bases in length. Physical analysis of the particles by absorbance and circular dichroic spectroscopy indicates that the DNA remains in the original conformation throughout the isolation procedure. Further, the particles reconstituted onto left-handed DNA compete effectively for an anti-Z DNA antibody, while the corresponding right handed particles do not. Analytical sedimentation velocity determinations indicate that the B-form poly[d(Gm5C)] and poly[d(GC)] particles sediment at 11.2 and 11.1 S, respectively. In contrast, the poly[d(Gm5C)] Z-form particles have an S20,w of 10.6 S. The differences in the sedimentation velocity and the density of the cores, and in the lengths of DNA associated with the particles, suggest that the conformation of the DNA affects the manner in which it associates with the histone octamer. PMID- 2986673 TI - Drug-protein interactions: binding of chlorpromazine to calmodulin, calmodulin fragments, and related calcium binding proteins. AB - The quantitative binding of a phenothiazine drug to calmodulin, calmodulin fragments, and structurally related calcium binding proteins was measured under conditions of thermodynamic equilibrium by using a gel filtration method. Plant and animal calmodulins, troponin C, S100 alpha, and S100 beta bind chlorpromazine in a calcium-dependent manner with different stoichiometries and affinities for the drug. The interaction between calmodulin and chlorpromazine appears to be a complex, calcium-dependent phenomenon. Bovine brain calmodulin bound approximately 5 mol of drug per mol of protein with apparent half-maximal binding at 17 microM drug. Large fragments of calmodulin had limited ability to bind chlorpromazine. The largest fragment, containing residues 1-90, retained only 5% of the drug binding activity of the intact protein. A reinvestigation of the chlorpromazine inhibition of calmodulin stimulation of cyclic nucleotide phosphodiesterase further indicated a complex, multiple equilibrium among the reaction components and demonstrated that the order of addition of components to the reaction altered the drug concentration required for half-maximal inhibition of the activity over a 10-fold range. These results confirm previous observations using immobilized phenothiazines [Marshak, D.R., Watterson, D.M., & Van Eldik, L.J. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 6793-6797] that indicated a subclass of calcium-modulated proteins bound phenothiazines in a calcium dependent manner, demonstrate that the interaction between phenothiazines and calmodulin is more complex than previously assumed, and suggest that extended regions of the calmodulin molecule capable of forming the appropriate conformation are required for specific, high-affinity, calcium-dependent drug binding activity. PMID- 2986674 TI - Complex formation and O2 sensitivity of Azotobacter vinelandii nitrogenase and its component proteins. AB - The O2 stability of the MoFe protein, the Fe protein, a 1:1 mixture of these proteins, and a 1:1 mixture in the presence of the Azotobacter vinelandii FeS-II protein has been studied as a function of time under controlled O2 partial pressures. The Fe protein is much more sensitive to O2 exposure than is the MoFe protein. The presence of the FeS-II protein at a 1:1 ratio with the component proteins measurably increases the O2 stability of the MoFe and Fe proteins. O2 inactivation of the MoFe protein was studied in some detail and found to be quite complex. At least three partially overlapping reactions are suggested. The first is the reversible oxidation of the metal clusters of the MoFe protein to the combined extent of 12 electrons with full retention of activity. The second phase consists primarily of activity loss with little increase in the extent of reversible oxidation. The third phase continues to decrease the protein activity but is also accompanied by formation of a g = 2.0 EPR signal and more extensive oxidation. Ultracentrifugation studies of the FeS-II protein at a 1:1:1 ratio with the Fe and MoFe proteins do not support the formation of the Bulen complex. The formation of other O2-stable complexes is discussed. PMID- 2986675 TI - Truncated repeated sequences generated by recombination in a specific region. AB - Structural relationships within a family of long repeated DNA sequences have been determined by molecular cloning of individual family members. About half of the family members are truncated at one end. There is a short, tandemly repeating region flanked by direct repeats associated with truncation. Recombination in a region near the tandemly repeating segment has apparently generated much of the diversity in this family. PMID- 2986676 TI - Phosphoenolpyruvate synthetase and pyruvate, orthophosphate dikinase: stereochemical consequences at both the beta-phospho and gamma-phospho groups of ATP. AB - [(R)-16O,17O,18O]Phosphoenolpyruvate and adenosine 5'-O-[(gamma S)-beta gamma 17O,gamma-17O,18O](3-thiotriphosphate) have been synthesized and used to determine the stereochemical course of the several displacements at phosphorus catalyzed by phosphoenolpyruvate synthetase and by pyruvate, orthophosphate dikinase, two enzymes that catalyze the formation of phosphoenolpyruvate from pyruvate and ATP. The catalytic mechanisms for each of these enzymes are believed to involve both phospho- and pyrophospho-enzyme intermediates. The stereochemical results are entirely in accord with these pathways: the beta-phospho group of ATP suffers overall retention of configuration that is presumably the consequence of two displacements with inversion, and the gamma-phospho group of ATP gamma S suffers inversion of configuration that is most probably the consequence of a single displacement at this center. PMID- 2986677 TI - Thrombin binds to a high-affinity approximately 900 000-dalton site on human platelets. AB - The functional sizes of the binding sites for thrombin on human platelets and isolated membranes have been determined by the technique of radiation inactivation: similar results were obtained. Independent studies using different radiation doses (0, 3, and 48 Mrad) and different thrombin concentrations (10( 10), 10(-8), and 10(-6) M) confirmed the presence of three binding sites with functional sizes of 900 000, 30 000, and 4000 daltons. The binding site of lowest apparent size (4000 daltons) probably corresponds to what has been termed nonspecific binding since its dissociation constant (2900 nM) is well outside the physiological range. The site of intermediate size (30 000 daltons) is also probably not involved in platelet activation since its dissociation constant (11 nM) is also beyond the concentration range required for activation, although it may be involved in other aspects of platelet-thrombin interaction. The sites with the largest functional size are probably important in platelet function since their dissociation constant (0.3 nM) is in the range required for platelet activation. The functional size of these sites (900 000 daltons) suggests that the high-affinity site for thrombin binding to platelets may involve a multimolecular complex of membrane components. PMID- 2986678 TI - Nucleotide sequence of a full-length complementary DNA clone and amino acid sequence of human phenylalanine hydroxylase. AB - A full-length human phenylalanine hydroxylase complementary DNA (cDNA) clone was isolated from a human liver cDNA library, and the nucleotide sequence encoding the entire enzyme was determined. The cDNA clone contains an inserted DNA fragment of 2448 base pairs, including 19 base pairs of poly(A) at the 3' end. The first methionine codon occurs at nucleotide position 223, followed by an open reading frame of 1353 base pairs, encoding 451 amino acids. Translation of the nucleotide sequence in the open reading frame predicts the amino acid sequence of human phenylalanine hydroxylase. The human protein shows a 96% amino acid sequence homology with the corresponding rat enzyme. The determination of the complete primary structure for phenylalanine hydroxylase represents the first among mixed-function oxidases. PMID- 2986679 TI - Membrane polypeptide in rabbit erythrocytes associated with the inhibition of L lactate transport by a synthetic anhydride of lactic acid. AB - The synthetic lactyl anhydride isobutylcarbonyl lactyl anhydride (iBCLA), a selective and potent inhibitor of L-(+)-lactate transport in rabbit erythrocytes, reduces the chemical labeling of a 40-50-kdalton polypeptide by tritiated 4,4' diisothiocyanato-2,2'-dihydrostilbenedisulfonate ([3H]H2DIDS). iBCLA does so in a dose-dependent manner at concentrations that strongly inhibit lactate lactate exchange but not chloride-phosphate exchange. These labeling experiments and inhibition reversal studies using iBCLA, p-(chloro-mercuri)benzenesulfonic acid (pCMBS), and dithiothreitol (DDT) suggest that iBCLA does not act at sulfhydryl groups but at or near an amino group that is near a disulfide linkage in the polypeptide which catalyzes lactate transport. These experiments support the association between specific monocarboxylate transport and a 40-50-kdalton membrane-bound polypeptide of the rabbit erythrocyte. PMID- 2986680 TI - Characterization of the fluorophore 4-heptadecyl-7-hydroxycoumarin: a probe for the head-group region of lipid bilayers and biological membranes. AB - The fluorophore 4-heptadecyl-7-hydroxycoumarin was used as a probe to study the properties of phospholipid bilayers at the lipid-water interface. To this end, the steady-state fluorescence anisotropy, the differential polarized phase fluorometry, and the emission lifetime of the fluorophore were measured in isotropic viscous medium, in lipid vesicles, and in the membrane of vesicular stomatitis virus. In the isotropic medium (glycerol), the probe showed an increase in the steady-state fluorescence anisotropy with a decrease in temperature, but the emission lifetime was unaffected by the change in temperature. In glycerol, the observed and predicted values for maximum differential tangents of the probe were identical, indicating that in isotropic medium 4-heptadecyl-7-hydroxycoumarin is a free rotator. Nuclear magnetic resonance and differential scanning calorimetric studies with lipid vesicles containing 1-2 mol % of the fluorophore indicated that the packaging density of the choline head groups was affected in the presence of the probe with almost no effect on the fatty acyl chains. The fluorophore partitioned equally well in the gel and liquid-crystalline phase of the lipids in the membrane, and the phase transition of the bilayer lipids was reflected in the steady-state fluorescence anisotropy of the probe. The presence of cholesterol in the lipid vesicles had a relatively small effect on the dynamics of lipids in the liquid-crystalline state, but a significant disordering effect was noted in the gel state. One of the most favorable properties of the probe is that its emission lifetime was unaffected by the physical state of the lipids or by the temperature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986681 TI - Kinetic mechanism for stimulation by monovalent cations of the amidase activity of the plasma protease bovine activated protein C. AB - A study of the effect of monovalent cations on the steady-state kinetic parameters for the hydrolysis of the synthetic substrate N alpha-benzoyl-L arginine-p-nitroanilide by activated bovine plasma protein C (APC) has been undertaken. The enzyme displayed a strict requirement for monovalent cations in its expression of amidolytic activity toward this substrate. Analysis of the variation in initial hydrolytic reaction rates, as a function of metal ion concentrations, suggested that at least two cation sites, or classes of sites, were necessary for catalysis to occur. After examination of the rate equations consequential to many different enzymic mechanisms that could account for these kinetic data, a mechanism was developed that fit the great majority of the experimental observations. In this mechanism it is postulated that cations bind to the enzyme in pairs, with a kinetically observable single binding constant, either preceded by or followed by binding of substrate. Catalysis occurs only after the enzyme-(metal cation)2-substrate complex is assembled. Some physical support for this mechanism was obtained upon the discovery that the binding (dissociation) constant for a competitive inhibitor of APC, p-aminobenzamidine, as determined by kinetic methodology, was independent of the concentration of Na+ and Cs+. PMID- 2986683 TI - Characterization of a Ca2+-calmodulin-stimulated cyclic GMP phosphodiesterase from bovine brain. AB - A calmodulin-stimulated form of cyclic nucleotide phosphodiesterase from bovine brain has been extensively purified (1000-fold). Its specific activity is approximately 4 mumol min-1 (mg of protein)-1 when 1 microM cGMP is used as the substrate. This form of calmodulin-sensitive phosphodiesterase activity differs from those purified previously by showing a very low maximum hydrolytic rate for cAMP vs. cGMP. The purification procedure utilizing ammonium sulfate precipitation, ion-exchange chromatography on DEAE-cellulose, gel filtration on Sephacryl S-300, isoelectric focusing, and affinity chromatography on calmodulin Sepharose and Cibacron blue-agarose results in a protein with greater than 80% purity with 1% yield. Kinetics of cGMP and cAMP hydrolysis are linear with Km values of 5 and 15 microM, respectively. Addition of calcium and calmodulin reduces the apparent Km for cGMP to 2-3 microM and increases the Vmax by 10-fold. cAMP hydrolysis shows a similar increase in Vmax with an apparent doubling of Km. Both substrates show competitive inhibition with Ki's close to their relative Km values. Highly purified preparations of the enzyme contain a major protein band of Mr 74 000 that best correlates with enzyme activity. Proteins of Mr 59 000 and Mr 46 000 contaminate some preparations to varying degrees. An apparent molecular weight of 150 000 by gel filtration suggests that the enzyme exists as a dimer of Mr 74 000 subunits. Phosphorylation of the enzyme preparation by cAMP-dependent protein kinase did not alter the kinetic or calmodulin binding properties of the enzyme. Western immunoblot analysis indicated no cross-reactivity between the bovine brain calmodulin-stimulated gGMP phosphodiesterase and the Mr 60 000 high affinity cAMP phosphodiesterase present in most mammalian tissues. PMID- 2986684 TI - Nucleotide sequence of a cDNA clone encoding mouse protamine 1. AB - The nucleotide sequence of a 404-base cDNA encoding the cysteine-rich, tyrosine containing mouse protamine has been determined. This insert, isolated from a mouse testis cDNA library, encodes a polypeptide of 50 amino acids of which 28 are arginine, 9 are cysteine, and 3 are tyrosine. The insert contains the complete 3' noncoding region of 151 bases and most of the 5' noncoding region. The predicted amino acid sequence of mouse protamine 1 is about 80% homologous to boar protamine and 67% homologous to bull protamine and contains the central, highly basic domain of four arginine clusters found in the trout protamines. The identification of a cDNA clone for a mouse protamine will facilitate studies of the evolution, regulation, and protein-DNA interaction of this nuclear protein unique to haploid spermatogenic cells. PMID- 2986682 TI - Specific arginine modification at the phosphatase site of muscle carbonic anhydrase. AB - Mammalian carbonic anhydrase III has previously been shown to catalyze the hydrolysis of p-nitrophenyl phosphate in addition to possessing the conventional CO2 hydratase and p-nitrophenylacetate esterase activities. Modification of pig muscle carbonic anhydrase III with the arginine reagent phenylglyoxal yielded two clearly distinctive results. Reaction of the enzyme with phenylglyoxal at concentrations equivalent to those of the enzyme yielded stoichiometric inactivation titration of the enzyme's phosphatase activity, approaching 100% loss of activity with the simultaneous modification of one arginine residue, the latter based on a 1:1 reaction of phenylglyoxal with arginine. At this low ratio of phenylglyoxal to enzyme, neither the CO2 hydratase activity nor the acetate esterase activity was affected. When the modification was performed with a significant excess of phenylglyoxal, CO2 hydratase and acetate esterase activities were diminished as well. That loss of activity was accompanied by the incorporation of an additional half dozen phenylglyoxals and, presumably, the modification of an equal number of arginine residues. The data in their entirety are interpreted to show that the p-nitrophenylphosphatase activity is a unique property of carbonic anhydrase III and that excessive amounts of the arginine modifying reagent lead to unspecific structural changes of the enzyme as a result of which all of its enzymatic activities are inactivated. PMID- 2986685 TI - Calcium-hydrogen exchange in isolated bovine rod outer segments. AB - We have measured Ca-H exchange in rod photoreceptors with different preparations of rod outer segments isolated from bovine retinas (ROS). One preparation contained ROS with an intact plasma membrane (intact ROS), and in the other preparation, the plasma membrane was leaky to small solutes (leaky ROS) and the cytoplasmic space was freely accessible to externally applied solutes. Addition of Ca2+ to Ca2+-depleted ROS (both intact and leaky) resulted in uptake of Ca2+ that was accompanied by the release of protons when catalytic amounts of the ionophore A23187 were present. This ionophore mediates Ca-H exchange transport across ROS membranes and serves to gain access to the intracellular compartment where Ca-H exchange appears to take place. Two protons were ejected for each calcium ion taken up. Conversely, when protons were added to Ca2+-enriched ROS, Ca2+ was released in the presence of A23187. The majority of this Ca-H exchange was observed only when A23187 was present in both intact and leaky ROS. We conclude that Ca-H exchange occurs predominantly in the intradiskal space and at the surface of the disk membrane rather than across the disk membrane. These exchange binding sites can accommodate 10 mol of Ca2+/mol of rhodopsin at physiological pH. We were unable to detect any Ca2+ release when a proton gradient was rapidly established across the disk membrane in the absence of A23187. These results are discussed in relation to the hypothesis that protons produced by the light-induced hydrolysis of cGMP cause the release of Ca2+ into the cytoplasm of rod photoreceptor cells. PMID- 2986686 TI - High-resolution proton nuclear magnetic resonance spectroscopy of chloride peroxidase: identification of new forms of the enzyme. AB - Chloride peroxidase from the mold Caldariomyces fumago in the native high-spin iron(III) and low-spin cyanoiron (III) states has been subjected to high-field proton nuclear magnetic resonance spectroscopic measurements. Signals shifted well outside the diamagnetic envelope by the paramagnetic iron(III) center are surprisingly insensitive to pH changes over the range from pH 3 to pH 7. The previously identified major form of chloride peroxidase (form A) and the minor form (B) show very similar chemical shift patterns. Of greatest significance, however, is the discovery that each of the separable forms of the enzyme exhibits splitting of porphyrin ring methyl resonances. The appearance of two sets of signals in both native and cyanide-complexed enzyme is best explained by the existence of two additional forms of the A and B isoenzymes. Structural differences for the newly identified forms of chloride peroxidase must be located in the vicinity of the heme prosthetic group. PMID- 2986687 TI - Probing the topography of lectins with site-specific spin-labeled glycosides. AB - Three new spin-labeled glycosides, spin-label I [1-[4-(beta-D galactopyranosyloxy)phenyl]-3-(2,2,6,6-tetramethyl-1 -oxypiperidin-4-yl)-2 thiourea], spin-label II (2,2,6,6-tetramethyl-1-oxypiperidin-4-yl alpha-D galactopyranoside), and spin-label III [1-(methyl 2-deoxy-alpha-D galactopyranosid-2-yl)-3-(2,2,6,6- tetramethyl-1-oxypiperidin-4-yl)-2-thiourea], were investigated as structural probes of Griffonia simplicifolia I isolectins (GS I) A4 and B4, respectively, by electron spin resonance (ESR) and inhibition of guaran isolectin precipitation. The p-aminophenyl beta-galactoside spin-label I was strongly immobilized by the B4 isolectin (Kd = 0.42 mM; 2T parallel = 54.0 +/- 0.3 G), while binding to the A4 isolectin was so weak (KI congruent to 2 mM) that binding was undetectable by ESR. The preference for the B4 isolectin was indicative of a more extended hydrophobic binding locus adjacent to the carbohydrate-specific binding site. The alpha-galactosyl spin-label II bound slightly more strongly to the A4 than to the B4 isolectin, as evidenced in both Kd values and particularly by differences in the degree of immobilization (2T parallel = 53.5 vs. 51.5 G, respectively). The 2-N-substituted methyl galactoside spin-label III was so poor an inhibitor of both isolectins (KI congruent to 1-2 mM) that ESR detection of the bound complex was not feasible. In all cases above, the spin-labels were displaced by specific monosaccharide haptens. PMID- 2986688 TI - Catabolism of bis(5'-nucleosidyl) oligophosphates in Escherichia coli: metal requirements and substrate specificity of homogeneous diadenosine-5',5'''-P1,P4 tetraphosphate pyrophosphohydrolase. AB - Diadenosine-5',5'''-P1,P4-tetraphosphate pyrophosphohydrolase (diadenosinetetraphosphatase) from Escherichia coli strain EM20031 has been purified 5000-fold from 4 kg of wet cells. It produces 2.4 mg of homogeneous enzyme with a yield of 3.1%. The enzyme activity in the reaction of ADP production from Ap4A is 250 s-1 [37 degrees C, 50 mM tris(hydroxymethyl)aminomethane, pH 7.8, 50 microM Ap4A, 0.5 microM ethylenediaminetetraacetic acid (EDTA), and 50 microM CoCl2]. The enzyme is a single polypeptide chain of Mr 33K, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and high-performance gel permeation chromatography. Dinucleoside polyphosphates are substrates provided they contain more than two phosphates (Ap4A, Ap4G, Ap4C, Gp4G, Ap3A, Ap3G, Ap3C, Gp3G, Gp3C, Ap5A, Ap6A, and dAp4dA are substrates; Ap2A, NAD, and NADP are not). Among the products, a nucleoside diphosphate is always formed. ATP, GTP, CTP, UTP, dATP, dGTP, dCTP, and dTTP are not substrates; Ap4 is. Addition of Co2+ (50 microM) to the reaction buffer containing 0.5 microM EDTA strongly stimulates Ap4A hydrolysis (stimulation 2500-fold). With 50 microM MnCl2, the stimulation is 900 fold. Ca2+, Fe2+, and Mg2+ have no effect. The Km for Ap4A is 22 microM with Co2+ and 12 microM with Mn2+. The added metals have similar effects on the hydrolysis of Ap3A into ADP + AMP. However, in the latter case, the stimulation by Co2+ is small, and the maximum stimulation brought by Mn2+ is 9 times that brought by Co2+. Exposure of the enzyme to Zn2+ (5 microM), prior to the assay or within the reaction mixture containing Co2+, causes a marked inhibition of Ap4A hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986689 TI - Covalent modification of both cAMP binding sites in cAMP-dependent protein kinase I by 8-azidoadenosine 3',5'-monophosphate. AB - Reconstituted porcine cAMP-dependent protein kinase type I was labeled with 8 azidoadenosine 3',5'-monophosphate (8-N3cAMP) to study cyclic nucleotide binding and to identify amino acid residues that are either in or in close proximity to the cAMP binding sites. The photoaffinity analogue 8-N3cAMP behaved as cAMP itself with respect to cyclic nucleotide binding. For both cAMP and 8-N3cAMP, 2 mol of nucleotide was bound per mole of type I regulatory subunit monomer (RI), the apparent Kd's observed were approximately 10-17 nM on the basis of either Millipore filtration assays, equilibrium dialysis, or ammonium sulfate precipitation, Scatchard plots showed positive cooperativity, and (4) the Hill coefficients were approximately 1.5-1.6. After photolysis and addition of an excess of cAMP, approximately 1 mol of 8-N3cAMP/mol of RI monomer was covalently incorporated. Tryptic digestion of the labeled protein revealed that two unique tryptic peptides were modified. Proline-271 and tyrosine-371 were identified as the two residues that were covalently modified by 8-N3cAMP in RI. These results contrast with the type II regulatory subunit (RII) where 8-N3cAMP modified covalently a single tyrosine residue [Kerlavage, A. R., & Taylor, S. S. (1980) J. Biol. Chem. 255, 8483-8488]. RI contains two adjacent regions of sequence homology in the COOH-terminal fragment that binds two molecules of cAMP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986690 TI - Distance between the visible copper and cytochrome a in bovine heart cytochrome oxidase. AB - Electron paramagnetic resonance (EPR) at 15 K was used to probe the magnetic interaction between the visible copper CuA2+ and ferric cytochrome a in the carbon monoxide compound of beef heart cytochrome oxidase. At pH 8.6, the midpoint potentials (Em's) for one-electron oxidation of CuA+ and cytochrome a2+ were found to be 195 and 235 mV, respectively. Because the Em of CuA is well below that of cytochrome a under these conditions, the microwave power saturation of CuA could be measured as a function of percentage cytochrome a oxidized. Although progressive power saturation data directly provide only the product of the spin-lattice and transverse relaxation rates delta [1/(T1T2)], Castner's theory for the saturation of inhomogeneously broadened lines [Castner, T.G., Jr. (1959) Phys. Rev. 115 (6), 1506-1515], along with our own theoretical formulation of the dipolar T2, enabled us to determine the change in T1 of CuA due to dipolar relaxation by cytochrome a. The orientation of the principal g values of CuA with respect to those of cytochrome a was evaluated in partially oriented membranous multilayers. When allowance was made for uncertainties in the relative CuA cytochrome a configuration and in the dipolar axis-magnetic field orientation, a range for the spin-spin distance r was calculated on the basis of the dipolar T1 of the gx component of CuA. This distance range was further restricted by consideration of T1 for the nonunique orientations of CuA giving rise to the gy signal. Only those values of r are possible for which the calculated T1 ratio (gx/gy) is equal to the experimentally determined ratio.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986691 TI - Soluble cytochrome composition of the purple phototrophic bacterium, Rhodopseudomonas sphaeroides ATCC 17023. AB - A detailed study of the soluble cytochrome composition of Rhodopseudomonas sphaeroides (ATCC 17023) indicates that there are five c-type cytochromes and one b-type cytochrome present. The molecular weights, heme contents, amino acid compositions, isoelectric points, and oxidation-reduction potentials were determined and the proteins were compared with those from other bacterial sources. Cytochromes c2 and c' have previously been well characterized. Cytochrome c-551.5 is a diheme protein which has a very low redox potential, similar to certain purple bacterial and algal cytochromes. Cytochrome c-554 is an oligomer, which is spectrally similar to the low-spin isozyme of cytochrome c' found in other purple bacteria (e.g., Rhodopseudomonas palustris cytochrome c 556). An unusual high-spin c-type heme protein has also been isolated. It is spectrally distinguishable from cytochrome c' and binds a variety of heme ligands including oxygen. A large molecular-weight cytochrome b-558 is also present which appears related to a similar protein from Rhodospirillum rubrum, and the bacterioferritin from Escherichia coli. None of the soluble proteins appear to be related to the abundant membrane-bound c-type cytochrome in Rps. sphaeroides which has a larger subunit molecular weight similar to mitochondrial cytochrome c1 and chloroplast cytochrome f. PMID- 2986692 TI - Phospholipid composition and organization of cytochrome c oxidase preparations as determined by 31P-nuclear magnetic resonance. AB - The molecular organization as well as the composition of the phospholipids in cytochrome c oxidase preparations (bovine heart) were investigated by 31P-nuclear magnetic resonance. In the so-called 'lipid-rich' preparation the lipids were found to form a fluid bilayer around the enzyme since the 31P-NMR spectrum was characteristic of a fast, axially symmetric motion of the phosphate groups with a chemical shift anisotropy of delta sigma = -45 ppm. In contrast, the 'lipid depleted' cytochrome c oxidase gave rise to a broader spectrum where the motion of the phospholipids was no longer axially symmetric. Nevertheless, the total width of the spectrum was still considerably narrower than observed for immobilized phospholipids in solid crystals. Both enzyme preparations were dissolved in 1% detergent solution and used for high-resolution 31P-NMR spectroscopy. Narrow lines of about 20 Hz linewidth were obtained for both types of enzyme preparations, and well-resolved resonances could be assigned to cardiolipin, phosphatidylethanolamin and phosphatidylcholine. The major differences between lipid-rich and lipid-depleted cytochrome c oxidase were the absolute amount of phospholipid associated with the protein and the relative contribution of the individual lipid classes to the 31P-NMR spectrum. For lipid rich cytochrome c oxidase about 130 molecules phospholipid were bound per enzyme (approx. 11 cardiolipins, 54 phosphatidylethanolamines and 64 phosphatidylcholines). For lipid-depleted cytochrome c oxidase only 6-18 lipids were bound per enzyme (1 or 2 cardiolipins, 3-8 phosphatidylethanolamines and 2-8 phosphatidylcholines). In contrast to earlier suggestions that cardiolipin is the only remaining lipid in lipid-depleted cytochrome c oxidase, the 31P-NMR studies demonstrate that all three lipids remain associated with the protein. PMID- 2986693 TI - Occlusion of Rb+ by detergent-solubilized (Na+ + K+)-ATPase from shark salt glands. AB - Occlusion of Rb+ by C12E8-solubilized (Na+ + K+)-ATPase from shark salt glands has been measured. The rate of de-occlusion at room temperature is about 1 s-1, which is the same as for the membrane-bound enzyme. The amount of Rb+ occluded is 3 moles Rb+ per mole membrane-bound shark enzyme, whereas only about 2 moles Rb+ are occluded by the C12E8-solubilized enzyme. PMID- 2986694 TI - Transport functions of the liver. Lack of correlation between hepatocellular ouabain uptake and binding to (Na+ + K+)-ATPase. AB - Ouabain uptake was studied on isolated rat hepatocytes. Hepatocellular uptake of the glycoside is saturable (Km = 348 mumol/l, Vmax = 1.4 nmol/mg cell protein per min), energy dependent and accumulative. Concentrative ouabain uptake is not present on permeable hepatocytes, Ehrlich ascites tumor cells and AS-30D ascites hepatoma cells. There is no correlation between ouabain binding to rat liver (Na+ + K+)ATPase and ouabain uptake into isolated rat hepatocytes. While ouabain uptake is competitively inhibited by cevadine, binding to (Na+ + K+)-ATPase is not affected by the alkaloid. Although the affinities of digitoxin and ouabain to (Na+ + K+)-ATPase are similar, digitoxin is 10000-times more potent in inhibiting [3H]ouabain uptake as compared to ouabain. That binding to (Na+ + K+)-ATPase appears to be no precondition for ouabain uptake was also found in experiments with plasmamembranes derived from Ehrlich ascites tumor cells and AS-30D hepatoma cells. While tumor cell (Na+ + K+)-ATPase is ouabain sensitive, the intact cells are transport deficient. Hepatic ouabain uptake might be related to bile acid transport. Several inhibitors of the bile acid uptake system also inhibit ouabain uptake. PMID- 2986695 TI - The effect of the intracellular sodium level on the activity of amino acid transport systems L and A in SV40 3T3 cells. AB - The rate of transport of phenylalanine and leucine, pertinent amino acids of System L, has been measured in SV40 3T3 cells as a function of the presence of Na+ ions during the reloading phase that precedes the influx determination. The presence of Na+ ions during the reloading phase resulted in an increase of the subsequent substrate influx through System L. This effect was related to the intracellular Na+ level and was found to be independent by the presence of a chemical sodium gradient outside-inside during influx determination; furthermore, this effect could not be ascribed to a difference between control and Na+-treated cells in the internal levels of those amino acids that participate in the exchange phenomena of transport System L. The transport of phenylalanine appeared to have the ability to accept Li+ for Na+ substitution in the 'trans' position. The presence of Na+ ions in the 'trans' position was not required to optimize the transport of System A-reactive substrates, whose influxes are dependent on the presence of the cation in 'cis' position. Analysis of the relationship between influx and substrate concentration indicated that the Na+-dependent increase of substrate influx was associated with an enlarged capacity of the high-affinity component of transport System L. PMID- 2986696 TI - Functional intron+ and intron- rDNA in the same macronucleus of the ciliate Tetrahymena pigmentosa. AB - Diallelic clones of Tetrahymena pigmentosa containing equal amounts of intron+ and intron- rDNA in the macronucleus were constructed. The macronucleus of the resulting strains divides amitotically during vegetative growth and the diallelic genotype is therefore unstable. The coexistence of the two alleles was followed in the total culture and in single cells during their vegetative segregation and it was observed that replication was non-preferential with respect to the two alleles. The diallelic clones were also used to demonstrate that intron containing rDNA was transcribed and the transcript processed in the presence of corresponding intron- rDNA. The results are discussed in the light of the 'non function' idea for ribosomal RNA introns. PMID- 2986698 TI - Cloning of a repeat sequence from human DNA which contains a BamHI site. AB - A human 1.5 kilobase BamHI repeated DNA fragment has been cloned from a genomic library and subcloned in pBR322. It is part of the human homogeneous main-band DNA, it has properties similar to those of long interspersed repetitive sequences (LINES), and differs from the families of human repeated DNA already described. PMID- 2986697 TI - IgG binding enhances DNAase I sensitivity of N-acetoxy-N-2-acetylaminofluorene modified phi X-174 RF DNA. AB - DNA restriction fragments of phi X-174 RF were modified with the carcinogen, N acetoxy-N-2-acetylaminofluorene (N-Aco-AAF). Immune complexes of 5'-32P-labeled AAF-modified DNA and rabbit immunoglobulin (IgG) against AAF-guanosine were specifically bound by surface membranes of Cowan I strain micrococci whose protein A binds the Fc portion of IgG. DNAase I sensitivity of the bound DNA was 20-fold greater than in solution, but the normal pattern of hydrolysis was not altered, as determined in sequencing gels. Nonadducted DNA ligated to AAF modified DNA acquired the enhanced sensitivity to DNAase I hydrolysis when the ligation hybrid was immunobound. PMID- 2986699 TI - A spectroscopic investigation of the structure and redox properties of Escherichia coli cytochrome b-562. AB - The six-coordinate monohaem ferricytochrome b-562 from Escherichia coli exhibits two haem-linked pH-dependent transitions detected by NMR and optical spectroscopy. Only one of these transitions, that of the Fe(III)-coordinated His 102, is detected by EPR and MCD; the ionisation of a haem propionate is not. Both ionisations are redox-state-dependent and the midpoint redox potential of the protein is markedly pH-dependent. Over the pH range 5.0 to 8.5 the potential drops from 260 mV to 110 mV and at least five single proton ionisations are responsible for this. In addition to the two spectroscopically identified ferricytochrome ionisations, there are at least three unidentified ionisations, two of which occur in the ferrous protein. From a consideration of the X-ray structure, together with NMR data, it seems probable that at least one of these ionisations involves an amino acid carboxylate. The X-ray structure also suggests that the relatively low pKa of His-102 is a result of its proximity to Arg-98. However, an appreciable interaction between these groups requires that the solution conformation differs slightly from the X-ray structure. The fast rate of electron self-exchange, over 4 X 10(6) M-1 X s-1 at 315 K and pH* 7, may be a reflection of the fact that, as shown by the X-ray structure, a large amount of the haem and axial histidine ligand are exposed at the molecular surface with an asymmetric distribution of charged groups surrounding them. PMID- 2986700 TI - The interaction of 5'-nucleotidase purified from chicken gizzard and actin, and the reversible loss of the inhibitory capacity of actin on deoxyribonuclease I. AB - Evidence is presented for a direct interaction of the intrinsic membrane protein 5'-nucleotidase (5'-ribonucleotide phosphohydrolase, EC 3.1.3.5) purified from avian smooth muscle (chicken gizzard) and the cytoskeletal component actin. Two different modes of interaction can be discerned: firstly, an immediate inhibitory effect of preferentially filamentous actin (F-actin) on the enzymic (i.e., AMPase) activity of 5'-nucleotidase and a direct binding of this enzyme to immobilized F-actin. Since these effects are suppressed by the addition of myosin subfragment 1, binding of 5'-nucleotidase appears to occur along the F-actin filament axis. Secondly, a time- and 5'-nucleotidase concentration-dependent transformation of also preferentially F-actin into a form unable to inhibit the enzymic activity of deoxyribonuclease I (DNAase I). This desensitization of actin versus DNAase I is not due to a denaturation process and was found to be reversible after addition of ATP. Furthermore, it does not seem to effect the ability of actin to bind to DNAase I. The transformation is accompanied by the hydrolysis of actin-bound nucleotide into adenosine, which remains bound to actin. Therefore, the desensitization of actin versus DNAase I appears to be due to a nucleotide-dependent conformational change of actin. An unidentified contamination of the 5'-nucleotidase preparations to a varying degree with ADPase and ATPase activities appears to be responsible for the desensitization process, although a synergistic role of these activities and 5'-nucleotidase cannot be excluded. PMID- 2986701 TI - Fucokinase, its anomeric specificity and mechanism of phosphate group transfer. AB - Fucokinase phosphorylates L-fucose at the anomeric position and, as such, might use either the alpha or beta anomer as its substrate. Examination of the utilization of radiolabelled alpha and alpha,beta mixtures established beta-L fucose as the required substrate. Phosphorylation at the anomeric center might involve either the loss or retention of the anomeric oxygen. The mechanism has been shown to involve anomeric oxygen retention through mass spectrometric analysis of the product phosphate derived from 18O-labelled L-fucose. PMID- 2986702 TI - Characterization of heme orientational disorder in myoglobin by proton nuclear Overhauser effects. AB - Freshly reconstituted sperm whale myoglobin is a mixture of two components distinguishable by proton nuclear magnetic resonance. The two species are interconvertible and the equilibrium composition is about 90% of one form, the form studied by X-ray methods. We have used the nuclear Overhauser effect to characterize the other (minor) component in its metcyano complex. Whereas in the major form there is dipolar contact between residue 99 and the heme pyrrole ring III, in the minor form the same residue is in contact with pyrrole IV, related to ring III by a 180 degrees rotation about the alpha-gamma meso axis. This interaction proves the validity of the heme rotational disorder proposition and confirms that the apoprotein does not discriminate between the two sides of the heme in the rapid insertion process. It is proposed that the differences in nuclear Overhauser effect between the protein matrix and the heme moiety can be used to define qualitatively the structural consequences of this heterogeneity. The altered heme-protein contacts could be related to the enhanced oxygen affinity in the minor form. PMID- 2986703 TI - Metabolism of cholesteryl ester in monolayers of bovine adrenal cortical cells. Effect of an inhibitor of acyl-CoA: cholesterol acyltransferase. AB - The effect of Sandoz compound 58-035 on cholesterol metabolism in monolayers of bovine adrenal cortical cells was studied. 58-035 did not inhibit cholesterol ester hydrolase, cholesterol side-chain cleavage, cholesterol synthesis from acetate, or cortisol synthesis in cells stimulated with ACTH or in unstimulated cells. It was, however, an effective inhibitor of formation of cholesteryl ester. The rate of formation of cholesteryl ester in the cells was increased by additional cholesterol derived from mevalonic acid or from the hydrolysis of intracellular lipid droplets. 58-035 caused an increase in the secretion of cortisol from cells maintained on a limited supply of cholesterol from bovine lipoproteins added to the medium when the cells were not stimulated with ACTH. This effect was not observed in stimulated cells. The results suggest that the bovine adrenal cortical cell can direct the flux of exogenous cholesterol very precisely according to its metabolic state. PMID- 2986705 TI - Stimulation of cholinephosphate cytidylyltransferase activity by estrogen in fetal rabbit lung is mediated by phospholipids. AB - We have investigated the mechanism by which estrogen stimulates phosphatidylcholine synthesis in fetal rabbit lung. The hormone increased the activity of cholinephosphate cytidylyltransferase in the 105 000 X g supernatant fraction but had no effect on the activities of this enzyme in the homogenate or other subcellular fractions. Although microsomal cytidylyltransferase has been reported to regulate phosphatidylcholine synthesis in other systems, and translocation of the enzyme from cytosol to microsomes has been reported in association with increased phosphatidylcholine synthesis, we found no evidence of this in the case of estrogen-stimulated phosphatidylcholine synthesis in the fetal lung. Cytosolic cytidylyltransferase activity was dependent on phospholipids. Extraction with acetone/butanol drastically reduced its activity as well as the stimulatory effect of estrogen. The activity and the effect of estrogen were restored on re-addition of lipids extracted with chloroform/methanol from additional supernatants. Fractionation of the total lipids revealed that the stimulatory effect was entirely associated with the phospholipids; neutral lipids and glycolipids did not stimulate. Treatment of the phospholipid fraction with phospholipase C abolished the stimulatory effect. The stimulatory effect of estrogen, however, could not be attributed to any individual phospholipid species but appeared to require the entire phospholipid mixture. We conclude that estrogen stimulates fetal lung phosphatidylcholine synthesis by increasing the activity of cytosolic cytidylyltransferase and this activation in turn is mediated by cytosolic phospholipids. PMID- 2986704 TI - Phospholipids, sterol carrier protein2 and adrenal steroidogenesis. AB - Rat adrenocortical cells and preparations of plasma membrane and mitochondria have been employed to assess the effects of phospholipids and of sterol carrier protein2 (SCP2) on specific aspects of adrenal steroidogenesis. With intact cells, liposomal dispersions of cardiolipin caused significant stimulation of corticosterone output, while preparations of phosphatidylcholine, phosphatidylinositol, or the 4'-phosphate and the 4',5'-diphosphate derivatives of phosphatidylinositol were without effect. With the adrenal plasma membrane preparation, none of the added phospholipids affected either sodium fluoride or ACTH-responsive adenylate cyclase activity. With intact mitochondria, only cardiolipin, among the various phospholipids, tested, caused a concentration dependent stimulation of pregnenolone production. However, even at the highest concentration of cardiolipin tested (500 microM), the stimulatory effect was only half that observed with 0.7 microM SCP2, and the two effectors were not synergistic. SCP2 caused a redistribution of cholesterol from mitochondrial outer to inner membranes, while cardiolipin, which is an activator of cytochrome P 450scc, had no effect on distribution of mitochondrial membrane cholesterol. PMID- 2986706 TI - Glycosphingolipids of leukemic cells in adult T-cell leukemia-lymphoma. AB - We analyzed lipids from leukemic cells of two patients with adult T-cell leukemia and compared them with those from T-cell lymphocytes of normal subjects. The neutral glycosphingolipids and gangliosides which were isolated were characterized by thin-layer chromatography and neuraminidase treatment. Both leukemic cells and normal lymphocytes had monoglycosylceramide and diglycosylceramide as major neutral glycosphingolipids. In one patient, diglycosylceramide was markedly increased. II3NeuAc-LacCer (GM3) and more complex gangliosides were detected in both cells. The most characteristic finding in leukemic cells was the occurrence of a disialylated ganglioside, II3(NeuAc)2 LacCer (GD3), which is not found in normal lymphocytes and neutrophils. This ganglioside may be due to the induced synthesis in association with malignant transformation. PMID- 2986707 TI - Cephaloridine-induced lipid peroxidation initiated by reactive oxygen species as a possible mechanism of cephaloridine nephrotoxicity. AB - Rat kidney microsomes reduced cephaloridine when incubated anaerobically with NADPH. Superoxide anion was generated in a concentration- and time-dependent manner when cephaloridine was incubated with rat kidney microsomes. Cephaloridine increased the in vitro peroxidation of rat kidney microsomal lipids in a concentration- and time-dependent manner. Cephaloridine-induced lipid peroxidation was inhibited by a combination of superoxide dismutase and catalase, by the hydroxyl radical scavengers, mannitol, (+)-cyanidanol-3 and by the singlet oxygen scavenger histidine in a concentration-dependent manner. It is proposed that cephaloridine nephrotoxicity may occur through the transfer of an electron from reduced cephaloridine to oxygen and subsequent formation of the superoxide anion, hydrogen peroxide, the hydroxyl radical and singlet oxygen. These activated oxygen species then are very likely to react with membrane lipids to induce lipid peroxidation and nephrotoxicity. PMID- 2986708 TI - Effect of administration of 2-methyl-4-dimethylaminoazobenzene on the half-lives of rat liver mitochondria and cytochrome oxidase. AB - The turnover of total mitochondrial proteins and cytochrome oxidase in the livers of rats administered with 2-methyl-4-dimethylaminoazobenzene (2-Me-DAB) has been determined. The incorporation of [14C]bicarbonate revealed a half-life of 3.1 days in control and 6 to 9 days in azodye administered animals for whole mitochondrial proteins. The incorporation of [35S]methionine yielded t1/2 values of 8.5 days and 15.4 days, respectively. The t1/2 of cytochrome oxidase, 10.8 days for control and 19.3 days for 2-Me-DAB-treated animals, indicated that the delay in the decay of the enzyme was of the same order as that of whole mitochondria. Short term incorporation revealed that the administration of the azodye stimulated the synthesis of the enzyme. Mitochondria isolated from azodye administered animals appeared less susceptible to lysosomal proteolysis. Also, azodye administration seemed to impair the ability of lysosomes to degrade mitochondria. PMID- 2986709 TI - Radical anions from one-electron-reduced adrenochrome. Detection and identification by electron spin resonance spectroscopy. AB - Free radicals from the one-electron reduction of adrenochrome have been studied in aqueous solutions. These radicals have been detected and identified by electron spin resonance spectroscopy, using spin stabilization methods (complexation with diamagnetic metal ions) to enhance radical concentrations. It is shown that the radicals have a characteristic ESR spectrum enabling their identification in complex systems. The spin density distribution in the radicals has been studied as a function of complexing metal ions and solvent composition. In the presence of oxidants (e.g., oxygen) the spectrum of the radical is replaced by that derived from the one-electron exidation of adrenochrome. PMID- 2986710 TI - A mouse tumor-derived osteolytic factor stimulates bone resorption by a mechanism involving local prostaglandins production in bone. AB - Culture medium which was conditioned by tissue of a CE mouse breast tumor in vitro contained dose-dependent osteolytic activity. The osteolytic activity was not soluble in dichloromethane and ethylacetate, indicating that it was not attributable to vitamin D metabolites or prostaglandins. However, breast tumor conditioned medium stimulated production and release of prostaglandin E2 from mouse calvaria in vitro, and the stimulation of bone resorption in vitro by breast tumor-conditioned medium was blocked by a dose of indomethacin that prevented stimulation of mouse calvarial prostaglandin E2 production and release. The resorptive activity of parathyroid hormone (PTH) was not affected by the same dose of indomethacin, suggesting that the osteolytic factor was not PTH. This was further supported by observation that mouse kidney cell cAMP production was stimulated by PTH, but not by the aqueous phase of ethylacetate-extracted breast tumor-conditioned medium. In addition to osteolytic activity, breast tumor conditioned medium contained a dose-dependent bone cell mitogenic activity, demonstrated by the stimulation of [3H]thymidine incorporation into trichloroacetic acid-insoluble macromolecules and a corresponding increase in bone cell number in monolayer cultures of bone cells. Breast tumor-conditioned medium also contained a dose-dependent transforming growth factor-(TGF-) like activity as defined by its ability to transform anchorage-dependent growth of nontransformed cells to anchorage-independent growth. The TGF in breast tumor conditioned medium did not compete with epidermal growth factor (EGF) for EGF receptor binding, but its transforming activity was greatly enhanced by EGF, indicating that it was a beta-type TGF. Both the osteolytic and mitogenic activities were nondialyzable, sensitive to reducing agent, and not removable by dichloromethane and ethylacetate extractions. Furthermore, the TGF activity was not removed by ethylacetate extraction. Thus, the possibility that these activities in breast tumor-conditioned medium might be mediated by the same molecule must be considered. In summary, our data suggest that the CE mouse mammary carcinoma cells produce and secrete into the culture medium an osteolytic factor which is neither PTH nor prostaglandin and which stimulates local synthesis in bone of prostaglandin E2 which in turn increases bone resorption in vitro. PMID- 2986711 TI - Isolation and kinetic properties of 5'-nucleotidase from guinea-pig skeletal muscle. AB - 5'-Nucleotidase (EC 3.1.3.5) has been solubilized and purified 1200-fold from guinea-pig skeletal muscle, to a specific activity of 40 U/mg protein. The purified enzyme yields a single protein band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Guinea-pig skeletal muscle 5'-nucleotidase is extremely sensitive to inhibition by nucleoside di- and triphosphates. The inhibition is of the competitive type, and can be reversed only by strong excess of Mg2+. Nucleoside diphosphates are more powerful inhibitors than nucleoside triphosphates. The Ki values for ADP and ATP are 0.036 and 0.28 microM, respectively. The purified enzyme does not require exogenous cations for maximal activity and is inhibited by EDTA. This inhibition is reversed by divalent cations. This indicates that the enzyme contains a tightly bound metal cation. PMID- 2986713 TI - Comparative reactivities of various biological compounds with myeloperoxidase hydrogen peroxide-chloride, and similarity of the oxidant to hypochlorite. AB - The reactivities of myeloperoxidase-H2O2-Cl- and sodium hypochlorite with amino acids, uric acid, NADH, ascorbic acid, ADP, albumin, haemoglobin, alpha 1 antitrypsin and some hydroxyl radical scavengers have been compared. The ability of each compound to inhibit chlorination of monochlorodimedon by both oxidants was measured. Relative reaction rates varied over a range of 10(5), but the reactivities of the two oxidants with each compound were very similar, from which it is concluded that the reactions of hypochlorite accurately reflect those of the myeloperoxidase system. Thiol compounds (cysteine and GSH) and methionine were more than 100-times more reactive than other amino acids, which had comparable reactivity to NADH and uric acid. Benzoate, dimethylsulphoxide and formate were very much less reactive. The significance of these reactions of myeloperoxidase in microbial killing and inflammation is discussed. PMID- 2986712 TI - Physicochemical studies of the iron(III)-carminomycin complex and evidence of the lack of stimulated superoxide production by NADH dehydrogenase. AB - Fe(III) complex of an antitumoral antibiotic carminomycin has been studied. Using potentiometric and spectroscopic measurements we have shown that carminomycin forms with Fe(III) a well-defined species in which three molecules of drug are chelated to one Fe(III) ion. This occurs with the release of one proton per molecule of drug. Magnetic susceptibility measurements suggest that six oxygen atoms are bound to iron. The stability constant is 3 X 10(34). The in vitro inhibition of P 388 leukemia cell growth by this complex compares with that of the free drug. This complex, unlike the free drug, does not catalyze the flow of electrons from NADH to molecular oxygen through NADH dehydrogenase. PMID- 2986714 TI - Triiodothyronine nuclear receptor and the role of non-histone protein factors in in vitro triiodothyronine binding. AB - The rat liver triiodothyronine (T3) nuclear receptor rapidly looses, after a partial purification from the nuclear extract, its ability to bind T3. We previously reported that histones, in the presence of DNA, could protect against inactivation enhancing the T3 binding site concentration and maintaining the high affinity for T3. A nuclear fraction discarded during the receptor purification (fraction A) was also found able to restore T3 binding and was analyzed. As histones + DNA, fraction A stabilized the T3 binding site from irreversible inactivation during incubation with T3, increasing its concentration while keeping the same high affinity for T3. It was active even at relatively high receptor concentration, appeared slightly more active than histones (+ DNA) in the same protein concentration range (up to 50-fold increment of T3 binding at the optimal concentration of 25 micrograms/ml) and was unaffected or slightly inhibited by DNA. Other proteins (ovalbumin, soybean trypsin inhibitor, RNAase) and rat liver cytosol were several times less effective, suggesting a major role of some nuclear constituents. The active factors in fraction A essentially belong to non-histone nuclear proteins. Fraction A was found heterogeneous regarding the molecular size and pHi of the active factors, the existence of subfractions more active on a protein concentration basis being suggested but not yet clearly evidenced. Efficient in vitro T3 binding to the isolated T3 nuclear receptor thus depends on the presence of several different nuclear constituents, histones + DNA or some non-histone proteins. Whether interactions with these constituents could modulate T3 binding within the nucleus remains to be elucidated. PMID- 2986715 TI - Purification and kinetic properties of polynucleotide kinase from rat testes. AB - Polynucleotide kinase (EC 2.7.1.78) has been purified from rat testes, and an approximately 2000-fold purification was obtained. The purified enzyme had an Mr of 38000 +/- 3800. The enzyme phosphorylated micrococcal nuclease-treated calf thymus DNA and (dT)10 while 5'-HO-tRNA was a very poor substrate. A certain degree of specificity towards purine-containing 5'-HO-nucleotides was observed. The polynucleotide kinase had an absolute requirement for a divalent cation. Both Mg2+ and Mn2+ could be used, but 10 mM MgCl2 gave optimal activity. The monovalent cations Na+, K+ and NH4+ all stimulated enzyme activity, and the optimal concentration was 0.1 M. The enzyme was inhibited by inorganic phosphate, pyrophosphate and sulphate. A 50% inhibition was obtained with 20, 0.3 and 2 mM, respectively. At 2 mM MgCl2, 1 mM spermine enhanced the enzyme activity 3-times. The apparent KATP was estimated to be 36 microM and KHO-DNA was found to be 2 microM. PMID- 2986717 TI - Subcellular localisation of inositol lipid kinases in rat liver. AB - The subcellular distribution of the enzymes which phosphorylate phosphatidylinositol sequentially to form phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate was investigated in rat liver. We demonstrate that whilst phosphatidylinositol kinase is present in Golgi, lysosomes and plasma membranes, the kinase that forms phosphatidylinositol 4,5 bisphosphate is localised predominantly at the plasma membrane. The role of the inositol lipid kinases in cell function is discussed. PMID- 2986716 TI - Abilities of some tryptophan and phenylalanine derivatives to inhibit gastric acid secretion. AB - Benzotript (N-p-chlorobenzoyl-L-tryptophan) has been shown to be a receptor antagonist in vivo and in vitro for peptides from the gastrin family. In the present study, we examine tryptophan, and some of its N- and C-acylated derivatives, as well as some phenylalanine derivatives, to show their ability to inhibit gastrin-induced acid secretion in the rat in vivo and to compete for the binding of [125I]-(Leu-15)-HG-17 to its cellular receptor on rabbit isolated gastric mucosal cells. N- and C- derivatives of tryptophan and phenylalanine were found to inhibit gastrin-induced acid secretion and binding of [125I]-(Leu-15)-HG 17 to its mucosal cell receptors. By either criterion, the relative antagonistic potencies of the compounds tested were: tert-butyloxycarbonyl-L-tryphophan-p nitrophenyl ester approximately equal to tert-butyloxycarbonyl-L-tryptophan carbamoylmethyl ester greater than tert-butyloxycarbonyl-L-tryptophyl-L-methionyl carbamoylmethyl ester approximately equal to tert-butyloxycarbonyl-L phenylalanine-carbamoylmethyl ester approximately equal to tert-butyloxycarbonyl L-tryptophyl-L-methionyl-amide greater than tert-butyloxycarbonyl-L-tryptophan greater than tert-butyloxycarbonyl-L-phenylalanine greater than benzyloxycarbonyl L-tryptophan approximately equal to benzotript, with minor differences between the in vivo and the in vitro experiments. These results demonstrate that both the nature of the amino acid residue and the N- and C-substitutions are important in determining antagonist activity and affinity for gastrin receptors. PMID- 2986718 TI - Differences in the reduction kinetics of incorporated spin labels in undifferentiated and differentiated mouse neuroblastoma cells. AB - Significant differences in the rate of reduction of two spin labels, 5 doxylstearic acid and TEMPOL, in the undifferentiated and differentiated NB-15 mouse neuroblastoma cells were demonstrated by using electron paramagnetic resonance (EPR) spectroscopy. The half-time (T1/2) values for decay of the EPR signal of 5-doxylstearic acid in the undifferentiated and differentiated neuroblastoma cells were 70 min and 290 min, respectively. The T1/2 values of TEMPOL in the undifferentiated and differentiated cells were 18 min and 34 min, respectively. The cellular reductant was characterized as non-protein-bound sulfhydryl groups. A corresponding difference in the cellular non-protein-bound sulfhydryl content, 19.30 nmol/mg protein for the undifferentiated cells and 6.78 nmol/mg protein for the differentiated cells, was observed. Comparison of the reduction rates of TEMPOL, 5-doxylstearic acid and 16-doxylstearic acid in the undifferentiated NB-15 cells suggested that the permeation of non-protein-bound sulfhydryl compounds from the cytosol to membrane may be responsible for the reduction of the lipid-soluble stearic acid spin labels. PMID- 2986719 TI - Inhibition by verapamil of neutrophil responses to formylmethionylleucylphenylalanine and phorbol myristate acetate. Mechanisms involving Ca2+ changes, cyclic AMP and protein kinase C. AB - Verapamil inhibits in human neutrophils the respiratory burst, the secretion and the change of transmembrane potential induced by formylmethionylleucylphenylalanine, a Ca2+-dependent stimulus, and by phorbol myristate acetate, a Ca2+-independent stimulus. Besides the blocking of Ca2+ channels, many mechanisms are responsible for the inhibition of neutrophil responses. In fact, verapamil (i) increases the intracellular cAMP concentration, potentiates the cAMP response induced by the chemotactic peptide and induces the appearance of a cAMP response also when the stimulant is phorbol myristate acetate; (ii) causes a decrease of Ca2+ association to cell membranes, so depleting the pools of exchangeable Ca2+ and depressing the 'Ca2+ response' in terms of rise in [Ca2+]i monitored with Quin 2 and of rapid mobilization from cell membranes monitored by chlorotetracycline fluorescence change; (iii) inhibits the Ca2+-activated phospholipid-dependent protein kinase C. The data, discussed in relation to the biochemical mechanisms of the stimulus-response coupling, are compatible with the hypothesis of an involvement of the activation of protein kinase C as key step in the sequence of transduction events for the induction of many neutrophil functions. PMID- 2986720 TI - The modulation of cell surface cAMP receptors from Dictyostelium discoideum by ammonium sulfate. AB - Dictyostelium discoideum cells contain a heterogeneous population of cell surface cAMP receptors with components possessing different affinities (Kd between 15 and 450 nM) and different off-rates of the cAMP-receptor complex (t 1/2 between 0.7 and 150 s). The association of cAMP to the receptor and the dissociation of the cAMP-receptor complex still occur in the presence of 3.4 M ammonium sulfate. However, these processes are strongly altered. (1) Low concentrations of ammonium sulfate (approximately equal to 50 mM) induce an approx. 2-fold increase of the number of cAMP binding sites. The same effect is induced by millimolar concentrations of CaCl2. Ammonium sulfate and CaCl2 are not additive, which suggests that these salts may act via the same mechanism. (2) High concentrations of ammonium sulfate (3.4 M) induce an alteration in the proportioning of the various cAMP binding sites to the components with the highest affinity. (3) High concentrations of ammonium sulfate (3.4 M) retard the dissociation of all binding sites about 3-6-fold, thus giving rise to an increase in the affinity of all cAMP binding components. PMID- 2986721 TI - Endocrine-mediated parallel changes in hepatic glucocorticoid and prolactin receptors. AB - The objective of this study was to determine whether in female rat liver any relationship existed between prolactin and glucocorticoid receptors after hormonal manipulation. Bromocryptine (CB-154) treatment of adult SD female rats (80-100 days old) for 48 h decreased prolactin binding to hepatic membranes 49% and dexamethasone binding in hepatic cytosol 40% below control values. Administration of rat prolactin along with bromocriptine prevented these changes. In another study, prolactin binding to hepatic membranes increased 53% and dexamethasone binding in hepatic cytosol increased 113% above sham-control values, 3 days after adrenalectomy. On the other hand, hydrocortisone treatment of sham-operated rats reduced prolactin binding by 57% and dexamethasone binding by 76%. Scatchard analyses of the prolactin or dexamethasone binding data indicated that these manipulations changed the number of prolactin or dexamethasone binding sites rather than their apparent affinity constants. In vitro treatment of rat whole liver homogenate with various doses (10(-9) - 10(-5) M) of dexamethasone and corticosterone for 15 min at 22 degrees C resulted in a dose-dependent decrease in prolactin binding activity. However, direct addition of dexamethasone to a hepatic 15 000 X g to 100 000 X g membrane preparation exhibited no significant effects on prolactin binding. In conclusion, these studies show that (a) there is a parallel in vivo modulation of rat liver prolactin and glucocorticoid receptors under various experimental conditions and (b) in vitro exposure of whole liver homogenate to glucocorticoids inhibits the prolactin binding activity. PMID- 2986722 TI - Stimulation of the hormone-sensitive triacylglycerol lipase from adipose tissue by phosphatidylethanolamine. AB - The activity of a pigeon adipose tissue hormone-sensitive triacylglycerol lipase preparation was increased from 2- to 5-fold by the presence of phosphatidylethanolamine in assays with three different methods of preparing triolein substrates. Phosphatidylethanolamine from egg yolk produced the greatest stimulation of lipase activity; the stimulation was concentration-dependent but was not time-dependent. A comparable increase in triacylglycerol lipase activity due to phosphatidylethanolamine was also observed with enzyme preparations from chicken and rat adipose tissue. Phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, cardiolipin, sphingomyelin, Triton X-100 and sodium dodecyl sulfate all inhibited enzyme activity. Phosphatidylethanolamine had no effect on acid lipase activity in the pigeon adipose tissue preparation. Preincubation of the pigeon adipose tissue lipase with ATP, cyclic AMP and protein kinase resulted in a 2.15-fold activation of hydrolase activity determined in the absence of phosphatidylethanolamine. In contrast, non-activated and protein kinase-activated forms of the lipase were characterized as having very nearly the same activity in assays with substrate preparations containing phosphatidylethanolamine. The phosphatidylethanolamine dependent stimulation of lipase activity was characterized kinetically as being due to an increase in maximal velocity. The modulation of the adipose tissue hormone-sensitive lipase activity by phospholipids could be involved in the hormonal regulation of lipolysis. PMID- 2986723 TI - Activation of oestrogen receptor complexes: evidence for the distinct regulation of ligand and oligonucleotide binding sites. AB - The activation of the rat uterine oestrogen receptor has been measured in vitro by its binding to oligodeoxythymidylate cellulose (oligo(dT] and was found to be sensitive to the time and temperature of prior incubation of cytosol with oestradiol. The presence of 20 mM dithiothreitol promoted receptor activation and was partially inhibited by 10 mM molybdate; molybdate also inhibited the time- and temperature-dependent activation of receptor. The nucleotides GTP, ATP, ADP, CTP and UTP all promoted receptor activation; the effect of GTP was significantly greater than that of ATP. It is unlikely that phosphate donation is involved in receptor activation as the effects of GTP could be reproduced by p[NH]ppG (guanosine 5'-[beta, gamma-imido]triphosphate), while PPi was also effective in activating receptor. The results provide evidence for the distinct regulation of the oligonucleotide- and ligand-binding domains, since manipulations which promoted binding to oligo(dT) did not affect either ligand binding capacity or the rate constant and composition the biphasic dissociation of the ligand receptor complex. PMID- 2986725 TI - The interconversion between monomeric and dimeric bovine heart cytochrome c oxidase. AB - Monomers and dimers of bovine heart cytochrome c oxidase (EC 1.9.3.1.) were separated by gel filtration chromatography on Ultrogel AcA 34 or by sucrose gradient centrifugation. Factors influencing the interconversion of the two aggregation states of this enzyme were analyzed. At very low ionic strength, in the presence of dodecyl maltoside, monomers were the main species. Salts appeared to stabilize the dimeric form, divalent cations being more efficient than monovalent. High enzyme concentrations favoured the formation of dimers, also at low ionic strength. The type of detergent had a strong influence on the monomer dimer interconversion; in Triton X-100 and dodecyl maltoside (at high ionic strength) cytochrome c oxidase was homogenously dispersed in its dimeric form, while in Tween-80 gel filtration showed only large particles eluting in the void volume. In cholate monomers and aggregates were observed but no dimers. The aggregation state had an influence on the steady state kinetics of the ferrocytochrome c oxidase activity. Monomers showed linear Eadie-Hofstee plots, whilst the dimeric and aggregated enzyme gave nonlinear Eadie-Hofstee plots. Ionic strength, enzyme concentration and type of detergent were affecting the enzyme's kinetics in a way consistent with the molecular form obtained by the gel filtration or sedimentation analysis. The data support a negative cooperative mechanism for the interaction of cytochrome c with the dimeric enzyme, as proposed earlier (K.A. Nalecz et al., (1983) Biochem. Biophys. Res. Commun., 114, 822-828). PMID- 2986724 TI - Inhibition of adipose S-100 protein release by insulin. AB - The release of S-100 protein brought about in rat epididymal fat pads by 10 microM epinephrine was inhibited by about 50% in the presence of more than 8 nM insulin. The inhibitory effect of insulin was also observed in the release of S 100 protein induced by isoproterenol or adrenocorticotropin (ACTH), but not in the release induced by a high concentration (5 mM) of dibutyryl cyclic AMP. Since insulin suppressed (to about 50%) the increase in cyclic AMP content induced by epinephrine under the same conditions, it is suggested that the inhibitory mechanism is mediated by the cyclic AMP levels in adipocytes. The S-100 protein release induced by catecholamine was significantly decreased (to about 50%) in the fat pads obtained from insulin-injected rats. In contrast, in the fat pads obtained from diabetic or long-term starved rats, the S-100 protein release was greatly enhanced, showing several-fold higher levels of basal release in the absence of hormones, and S-100 protein contents in the epididymal adipose tissues of these rats were significantly lower than those of the control rats. These results suggest that the S-100 protein content in adipocytes is regulated by insulin as well as the lipolytic hormones. PMID- 2986726 TI - [The Na+/H+ exchanger in eukaryotic cells: biochemical and pharmacological properties and physiological role]. AB - A membrane mechanism that catalyses the electroneutral exchange of Na+ for H+ has recently been characterized in a variety of eukaryotic cells. This exchanger is inhibited by amiloride, a potent diuretic drug. It has been implicated in a number of important physiological processes such as the regulation of the intracellular pH, the reabsorption of Na+ by the renal proximal tubule, the regulation of the cell volume and the fertilization of the sea urchin egg. The Na+/H+ exchanger seems able to mediate the action of growth factors. The biochemical and pharmacological properties of the Na+/H+ exchange system are reviewed. They are very similar in the different cell types that have been studied. Yet the Na+/H+ exchange system can fulfil different functions in different cell types depending i) on its properties of interaction with intracellular H+, ii) on the presence of other membrane structures that are involved in the maintenance of transmembrane Na+ and H+ gradients and iii) on the presence of extracellular messages that modify its catalytic properties and, among them, its interaction with internal H+. PMID- 2986727 TI - Indirect effects of the 3'-5' cyclic adenosine monophosphate binding protein (CAP) on the transcription of the malPQ operon in Escherichia coli. AB - Uninduced malPQ transcription, as followed by measuring beta-galactosidase expression in a strain carrying a malP-lacZ hybrid gene and grown in the absence of maltose, requires the presence of CAP. However this requirement is lost when the expression of malT, positive regulator gene of the maltose regulon, is rendered independent of CAP by a mutation in the malT promoter. This result suggests that the effect of CAP on uninduced malPQ expression is mediated through a modulation of MalT protein synthesis. The effect of CAP on the induced expression of malPQ is presumably mediated, in addition, through a modulation of the synthesis of the maltose transport system and, hence, of the entry of the inducer. Therefore the effect of CAP on malPQ expression seems to be merely indirect, and this is surprising since a CAP binding site is present at the malPQ promoter. PMID- 2986729 TI - Catabolite repression 1985. AB - The present status of catabolite repression is summarized with respect to the involvement of cyclic AMP and other mediators. A model is presented which may account for the relationship between positive control of gene expression exerted by cAMP and its receptor, CAP, and negative control of catabolite repression mediated by specific metabolites. PMID- 2986728 TI - Specific binding of the cAMP receptor protein of Escherichia coli to the lactose operon promoter. AB - The nitrocellulose filter binding assay has been used to study effects of pH, temperature, ionic strength and magnesium ions on the specific binding of the cyclic adenosine 3',5'-monophosphate (cAMP) receptor protein (CAP) to the promoter of the lactose (lac) operon of Escherichia coli. The pH has a significant effect on binding with the greatest amount of specific binding appearing at pHs near 7 with a gradual decrease in binding as the pH is increased to 8. Specific binding was observed at temperatures of 22 degrees C and 37 degrees C but not at 4 degrees C. The specific binding was also found to be a function of the concentration of magnesium acetate and potassium chloride, being dependent on the specific cation present, the total ionic strength, and the concentration of the CAP protein. All binding decreases as the ionic strength, increases, but this decrease occurs at a lower ionic strength in magnesium acetate than in potassium chloride. In a double label experiment the filter assay demonstrates that the cAMP-CAP complex preferentially binds to the wild-type lac promoter in the presence of a lac promoter mutated at the CAP binding site. Based on these results and comparisons with other experiments reported in the literature, buffer conditions that approximate the physiological state of a cell appear to be best for studying the interaction between CAP and the lactose promoter in vitro. PMID- 2986730 TI - [Control of the cell division cycle and sporulation in Saccharomyces cerevisiae by the cyclic AMP system]. AB - This paper reviews recent data on the adenylate cyclase system of the yeast Saccharomyces cerevisiae. Since the discovery of yeast adenylate cyclase mutants and the possibility of molecular biological analysis, adenylate cyclase and the subsequent steps in the cAMP cascade have become subject of intense investigation. CYR1, the structural gene for the adenylate cyclase catalytic subunit is necessary for cell division and in diploid cells is involved in the choice between sporulation and cell division. The cell division cycle in yeast is initiated by a step called START, which has been defined by mutations causing an arrest of the cells in an unbudded state. One class of mutation causes the cell to arrest at the same stage of the cell division cycle as the pheromone implicated in conjugation. A second class causes cells to cease growth in a different manner, but one which is similar to the arrest brought about by nutient deprivation. The adenylate cyclase gene belongs to the second class and has been identified as CDC35. Two genes of the first class have been cloned and sequenced. CDC28 codes for a kinase which has homology with the src proto-oncogene family. CDC36 is partly homologous with the oncogene ets. Two genes related to the ras oncogene family have also been implicated in the control of START. START can be dissociated in two subsequent phases, the first being controlled by the AMPc system and the second including proto-oncogenes. A model in which cAMP is a positive indicator of available nutrients such as nitrogen has been constructed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986731 TI - Recent aspects of genetic manipulation in Bacillus thuringiensis. AB - The conjugative plasmid pAM beta 1 was transferred from Streptococcus faecalis to several strains of Bacillus thuringiensis by a filter-mating process. From a transconjugant clone of B. thuringiensis a hybrid plasmid resulting from an in vivo insertion into pAM beta 1 of a 3 Md DNA sequence was isolated. This 3 Md DNA molecule (Th sequence) is related to several host plasmids found in different serotypes of B. thuringiensis. A reciprocal conjugation-like process involving the transfer of pAM beta 1 from B. thuringiensis to S. faecalis was also demonstrated. The comparison of the restriction maps of the crystal genes from plasmid and chromosomal origins of different serotypes, six of which having been cloned in E. coli, revealed the existence of two classes of genes which are very similar in the map corresponding to the N-terminal part of the protein, and which differ essentially in the 3' region. The presence of the transposon-like Th sequence was found in several cases associated with the crystal gene in the same host plasmid, and a model for their structural organization is proposed. PMID- 2986732 TI - [Activity of cAMP-dependent protein kinases and cAMP-binding proteins from rat renal cytosol upon dehydration]. AB - The activity of cAMP-dependent protein kinases, cAMP binding and the spectrum of cAMP-binding proteins in renal papillary cytosol of intact rats and of rats kept on a water-deprived diet for 24 hours were investigated. It was found that the stimulation of protein kinases by 10(-6) M cAMP in the experimental group was significantly higher than in the control one. On DEAE-cellulose chromatography, the position of peaks of the specific cAMP binding corresponded to those of the regulatory cAMP-dependent protein kinases type I and II. Under these conditions, more than 80% of the binding activity in intact animals was localized in peak II, whereas in rats kept on a water-deprived diet over 60% of the binding activity was localized in peak I. The total binding activity of cytosol in experimental animals remained unchanged is compared to intact rats. It is suggested that in renal papilla dehydration is accompanied by the induction of synthesis of regulatory subunits of cAMP-dependent protein kinase type I. PMID- 2986733 TI - [Heterologous desensitization of pigeon erythrocyte adenylate cyclase by the catalytic subunit of cAMP-dependent protein kinase]. AB - Preincubation of pigeon erythrocyte plasma membranes with the catalytic subunit of cAMP-dependent protein kinase results in the desensitization of erythrocyte adenylate cyclase. The adenylate cyclase activity measured in the presence of 10 microM isoproterenol and 50 microM GTP-gamma-S decreases by 40% after 10 min incubation; that in the presence of 50 microM GTP-gamma-S by 35% (20 min). The decrease of the adenylate cyclase activity is due to the prolongation of the lag phase of the enzyme activation in the presence of a hydrolysis-resistant GTP analog and to the drop in activity in the steady state of the activation. The heterologous desensitization of adenylate cyclase induced by cAMP-dependent protein kinase is also coupled with the decrease of the number of beta adrenoreceptors capable of acquiring a high affinity for the agonists in the absence of guanyl nucleotides. The effect of the catalytic subunit on adenylate cyclase is fully compatible with the process of the enzyme desensitization in erythrocytes treated with isoproterenol or cAMP. PMID- 2986734 TI - [Polymorphism of membrane proteinases from mitochondria]. AB - Using native polyacrylamide gel electrophoresis in the absence of sodium dodecyl sulfate, the detergent extracts of sonic submitochondrial particles (SMP) were separated into three protein fractions capable of accomplishing the proteolysis of cytochrome c and three other fractions catalyzing the hydrolysis of N-a benzoyl-L-arginine-p-nitroanilide (BAPA) and N-a-benzoyl-L-arginine-B naphthylamide (BANA). The fractions isolated from the gel were subjected to a thorough anaylsis. Cytochrome c hydrolases were shown to have identical molecular weights (17000) but different isoelectric points (4.0, 4.2 and 4.4). The total cytochrome c hydrolase activity of these enzymes was inhibited by phenylmethylsulfonylfluoride but was insensitive to ethylenediaminetetraacetate and o-phenanthroline. Three BANA (BAPA) hydrolases have identical Mr values (approximately 17500) but different pI values (4.2, 4.3 and 4.7). Apart from the indicated hydrolases, the detergent extracts of SMP were shown to contain minor components with identical activities distinguished by the tightness of binding to the inner mitochondrial membrane, Mr and sensitivity to proteinase inhibitors. The observed phenomenon is considered to be due to the polymorphism of proteinases coupled with the inner mitochondrial membrane. PMID- 2986735 TI - [Changes in the physico-chemical properties of synaptosomal membranes induced by phospholipase A2]. AB - Using fluorescent and EPR spin probing techniques, the changes in the physico chemical properties of rat brain synaptosomal membranes induced by phospholipase A2 were studied. It was shown that treatment of synaptosomal membranes with phospholipase A2 leads to their depolarization, increases their surface negative potential and decreases the microviscosity of the membrane lipid bilayer. The observed changes in the physicochemical properties of synaptosomal membranes induced by phospholipase A2 are discussed in terms of a possible regulatory role of lipids in the transmembrane chemical signal transfer. PMID- 2986736 TI - Adrenocorticotrophic hormone in the anterior and neurointermediate lobes of the rat during the perinatal period: polymorphism, biological and immunological activities of ACTH. AB - When acid extracts (0.1 N HCl) of the anterior or neurointermediate lobes of fetal and newborn rat pituitary were subjected to gel filtration on Sephadex G-50 fine columns, polymorphism of ACTH was revealed by radioimmunoassay. At every stage of pregnancy and after birth three main ACTH forms were observed. The first form ('big' ACTH) eluted near the void volume of the column; its apparent molecular weight was 45,000. The second form ('little' ACTH) coeluted with synthetic human ACTH1-39 and 125I human ACTH. As the latter marker eluted more slowly than the former, 'little' ACTH, which showed an apparent molecular weight near 5,000, could not be a single entity. The third form ('intermediate' ACTH) eluted between 'big' and 'little' ACTH; its apparent molecular weight was near 13,000. During the perinatal period, the relative proportions between these different molecular forms of ACTH changed in both anterior and neurointermediate lobes. Thus the gradual increase of 'intermediate' and 'little' forms was accompanied by a decrease of 'big' ACTH. The perfusion of fetal adrenal glands with the same immunological quantities of 'big', 'intermediate' and 'little' ACTH showed that the 'little' and 'intermediate' forms have greater biological potency in eliciting corticosterone secretion than the 'big' ACTH form. PMID- 2986737 TI - Hormone response to repeated electroconvulsive therapy: effects of naloxone. AB - Plasma prolactin (PRL), cortisol, and growth hormone (GH) were measured before, and at 15-min intervals for 1 hr after, electroconvulsive therapy (ECT). This was repeated over a series of 6 consecutive treatments for each of 12 depressed drug free inpatients. Patients received naloxone, 2 mg or 20 mg, by intravenous infusion before the third and fifth treatment. ECT was consistently followed by a release of PRL and cortisol, although two patterns of PRL response could be distinguished. In eight patients, the PRL response did not change significantly with repeated ECT, whereas in four patients, the plasma PRL increased tenfold after the first treatment and decreased after each successive treatment. The GH level varied widely, with no evidence of a reliable response to ECT. Opiate receptor blockade with low- or high-dose naloxone did not alter the release of PRL or cortisol after ECT. These findings demonstrate a reliable PRL and cortisol response to ECT, but do not support a role for endogenous opiates in these hormonal changes. PMID- 2986738 TI - Evidence for a role for cellular alkalinization in the cyclic adenosine 3',5' monophosphate-mediated initiation of motility in bovine caput spermatozoa. AB - Bicarbonate ion, the local anesthetics procaine and dibucaine, and the ionophores monensin and nigericin have been shown to markedly increase the ability of agents that elevate cyclic adenosine monophosphate (cAMP) levels to initiate motility in bovine caput spermatozoa. A number of other weak bases, including theophylline, D 600 and dipyridamole, elevate cAMP levels maximally in caput sperm at low levels but induce motility only at high levels. These compounds thus appear to have a dual role in the initiation of motility, i.e., they elevate both cAMP levels and internal pH. Confirmation of this view was provided by the demonstration that bicarbonate ion and procaine permit initiation of motility by theophylline, D-600 and dipyridamole at markedly reduced levels. Also, forskolin (a neutral adenylate cyclase activator) elevates cyclic AMP levels in caput sperm but initiates motility only in the presence of bicarbonate or procaine, and the membrane permeant cAMP analogue 8-bromo-cAMP is capable of inducing motility only in the presence of bicarbonate. Thus, motility in caput sperm is induced only under conditions that elevate both intracellular cAMP and pH, whereas caudal sperm motility is stimulated by an elevation of either cAMP or pH. These data suggest that the epididymal development of motility requires a maturational increase in internal pH. This suggestion was confirmed by direct measurement of the internal pH of caput and caudal sperm; the internal pH of the former was found to be 5.84 +/- 0.1 and the latter 6.27 +/- 0.05. PMID- 2986739 TI - Binding of high-density lipoproteins to luteal membranes: the role of prolactin, luteinizing hormone, and circulating lipoproteins. AB - Ovarian and adrenal membranes from immature gonadotropin-primed rats, treated with 4-amino-pyrazolopyrimidine (4APP) to reduce endogenous lipoprotein levels, displayed higher binding of porcine high-density lipoprotein (HDL) when compared to control rats. Immature, hypophysectomized (HYPOX) rats bearing corpora lutea (CL) on Day 5 after ovulation had lower levels of serum progesterone and reduced capacity for HDL and human chorionic gonadotropin (hCG) binding to ovarian membranes when compared with intact animals. Hypophysectomy also reduced the number of HDL binding sites in adrenal membranes. Treatment of HYPOX animals with luteinizing hormone (LH) and prolactin (Prl) alone or in combination increased the HDL binding sites in the ovary relative to HYPOX-untreated rats. Neither hormone affected binding to adrenals, where only adrenocorticotropic hormone (ACTH) enhanced HDL binding. LH treatment reduced the serum progesterone levels and hCG binding to the ovaries, whereas Prl administration increased progesterone levels with no effect on hCG binding. We conclude from this study that HDL binding in the luteinized ovary is regulated by Prl and LH and circulating lipoproteins, whereas in adrenals it is regulated by ACTH and circulating levels of lipoproteins. PMID- 2986740 TI - Pituitary luteinizing hormone-releasing hormone receptors in ovariectomized cows after challenge with ovarian steroids. AB - Acute changes of bovine pituitary luteinizing hormone-releasing hormone (LHRH) receptors in response to steroid challenges have not been documented. To investigate these changes 96 ovariectomized (OVX) cows were randomly allotted to one of the following treatments: 1) 1 mg estriol (E3); 2) 1 mg 17 beta-estradiol (E2); or 3) 25 mg progesterone (P) twice daily for 7 days before 1 mg E2 and continuing to the end of the experiment. Serum was collected at hourly intervals from 4 animals in each group for 28 h following estrogen treatment. Four animals from each treatment were killed at 4-h intervals from 0 to 28 h after estrogen injection to recover pituitaries and hypothalami. Treatment with E3 or E2 decreased serum luteinizing hormone (LH) within 3 h and was followed by surges of LH that were temporally and quantitatively similar (P greater than 0.05). Progesterone did not block the decline in serum LH, but did prevent (P less than 0.05) the E2-induced surge of LH. Serum follicle-stimulating hormone (FSH) was unaffected (P less than 0.05) by treatment. Pituitary concentrations of LH and FSH were maximal (P less than 0.001) at 16 h for E3 and 20 h for E2, whereas P prevented (P greater than 0.05) the pituitary gonadotropin increase. Concentrations of LHRH in the hypothalamus were similar (P greater than 0.05) among treatments. Pituitary concentrations of receptors for LHRH were maximal (P less than 0.005) 12 h after estrogen injection (approximately 8 h before the LH surge), even in the presence of P. This study demonstrated that in the OVX cow: 1) E2 and E3 increased the concentration of receptors for LHRH and this increase occurred before the surge of LH; and 2) P did not block the E2-induced increase in pituitary receptors for LHRH but did prevent the surge of LH. PMID- 2986741 TI - Epstein-Barr virus-associated hemophagocytic syndrome: virological and immunopathological studies. AB - The virus-associated hemophagocytic syndrome (VAHS) is a disorder characterized by a benign, generalized histiocytic proliferation, with marked hemophagocytosis associated with systemic viral infections. We have studied the virological and immunopathological events occurring in two children experiencing Epstein-Barr VAHS. Neither of the patients had an underlying immunodeficiency and both recovered from their disease and are completely well one year after follow-up. In each patient, evidence for primary Epstein-Barr virus (EBV) infection was documented with a typical humoral immune response, including IgM antibody directed against virus capsid antigen. EBV was demonstrated in lymphoreticular tissues by electron microscopy and molecular hybridization studies. Permissive EBV infection was suggested by the finding of mature virus particles and linear viral DNA in lymphoreticular tissues. Immunopathological studies demonstrated complete effacement of lymph node architecture by a marked proliferation of immunoblasts in patient 1 and infiltration and effacement of the lymph node architecture with benign-appearing histiocytes in patient 2. Atypical lymphocytes characteristic of acute EBV infection were notably absent in the peripheral blood of both patients and cytotoxic T cells, which normally lyse EBV-infected B cells, were also absent from the peripheral circulation. Our observations suggest that EBV-induced VAHS may be the result of an increased virus burden in the face of immunoregulatory cell imbalances. PMID- 2986742 TI - Donor cell leukemia developing six years after marrow grafting for acute leukemia. AB - A patient who developed recurrent leukemia more than six years after marrow grafting from an HLA-identical same-sex sibling is reported. Difference in DNA restriction fragment length polymorphisms between donor and host demonstrated that the DNA in the recurrent leukemia sample was probably of donor origin. Possible mechanisms that could explain the long latent period between transplantation and expression of leukemic transformation are discussed. We conclude that future cases of late leukemic recurrence after marrow grafting should be studied to determine whether, in contrast to early relapses, late relapses occur in donor cells in most or all instances. PMID- 2986743 TI - Abnormal adherence-related functions of neutrophils, monocytes, and Epstein-Barr virus-transformed B cells in a patient with C3bi receptor deficiency. AB - We evaluated a 3-year-old female patient with leukocytosis, recurrent infections, severe periodontal disease, and a history of delayed separation of the umbilical stump. This patient's polymorphonuclear leukocytes (PMNs) had normal membrane depolarization responses, normal oxygen metabolism, normal granule secretion responses, normal bactericidal activity, and normal C3b rosetting. However, by fluorescent cell analysis and C3bi rosetting, it was determined that her cells lacked the C3bi receptor. In addition, the patient's PMNs showed markedly abnormal chemotaxis, adherence, and aggregation responses, and partial abnormalities were detected in PMN spreading and phagocytosis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the subject's neutrophil cytoplasts were missing a 180,000-dalton moiety. Her monocytes also had defective chemotaxis and failed to adhere and grow normally in culture. Epstein-Barr virus transformed B cells from the patient lacked an aggregation response to phorbol myristate acetate. Laboratory and clinical evaluations of this patient's mother showed no abnormalities. These studies demonstrate that C3bi receptor deficiency can be associated with functional abnormalities in multiple myeloid cells and that the absence of C3bi receptor is associated with abnormal adherence-related functions of these cells. PMID- 2986744 TI - Adult T cell leukemia: a potential target for ricin A chain immunotoxins. AB - Adult T cell leukemia (ATL) is an almost uniformly fatal malignancy of mature T cells associated with human T cell leukemia/lymphoma virus type 1 (HTLV-1) infection. Cells from this leukemia are characterized by the expression of large numbers of receptors for interleukin 2 (IL-2). In an attempt to prepare an immunotoxin with selective cytotoxicity for ATL cells, we conjugated anti-Tac, a monoclonal anti-IL-2 receptor antibody, to purified ricin A chains. Although unmodified anti-Tac had no effect on the protein synthesis of these cells, anti Tac-ricin A chain conjugates produced half-maximal inhibition of protein synthesis in HTLV-1-infected leukemic T cell lines at concentrations of 2 to 6 X 10(-10) mol/L (ID50). An essentially identical ID50 was obtained with leukemic peripheral blood T lymphocytes isolated from two patients with ATL. In contrast, half-maximal inhibition of protein synthesis in HTLV-uninfected, IL-2 receptor negative T and B cell lines required 200- to 1,000-fold higher concentrations of anti-Tac-ricin A chain conjugates. Both unconjugated anti-Tac and immunoaffinity purified IL-2 completely inhibited the toxic effects of anti-Tac-ricin A, confirming the specificity of the conjugate-IL-2 receptor interaction. Clonogenic assays demonstrated that anti-Tac-ricin A chain was able to eliminate greater than 99.9% of an HTLV-1-infected T cell population at concentrations only marginally affecting IL-2 receptor-negative cells. The data presented demonstrate that anti-Tac-ricin A is selectively cytotoxic for HTLV-1-infected leukemic T cells in vitro and raises the future possibility of specific therapeutic intervention with immunotoxins in this disease. PMID- 2986746 TI - Alpha zero-thalassemia due to recombination between the alpha 1-globin gene and an AluI repeat. AB - A form of alpha zero-thalassemia found in subjects of Mediterranean origin has been analyzed by gene mapping and DNA sequencing. Homozygotes have the hemoglobin Bart's hydrops fetalis syndrome, while compound heterozygotes for this defect and alpha+-thalassemia have hemoglobin H disease. It results from a deletion that removes 20.5 kilobases of DNA from within the alpha-globin gene cluster. Sequence data from the regions adjacent to the breakpoint indicate that the recombination event that caused this deletion occurred between the alpha 1-gene and an unusual AluI sequence located between the embryonic zeta genes. PMID- 2986745 TI - The regulation of immunoglobulin synthesis after HLA-identical bone marrow transplantation: VI. Differential rates of maturation of distinct functional groups within lymphoid subpopulations in patients after human marrow grafting. AB - This investigation uses different polyclonal activators of in vitro immunoglobulin production to elicit immunoregulatory profiles of B cells, T cells, T4 cells, and T8 cells from 25 recipients (13 with and 12 without chronic graft-v-host disease [GVHD] ) after HLA-identical marrow transplantation for aplastic anemia or hematologic malignancy. Pokeweed mitogen, Epstein-Barr virus, herpes simplex type 1 virus, and tetanus toxoid were used to induce immunoglobulin production as measured by a plaque assay. Multiple defects in the various lymphoid subsets were found in both groups of patients. There was defective b cell function, lack of T cell or T4 cell subset helper activity, and increased T cell, T4 cell, or T8 cell suppressor activity after stimulation with the various activators. Inconsistent B, T, T4, and T8 cell functions in the marrow graft recipients provide evidence for (a) different functional groups of cells within each subset responsive to different polyclonal activators; (b) a spectrum of immune capabilities within each phenotype lineage; (c) different patterns of immune reconstitution for each lymphocyte subset after marrow grafting; and (d) chronic GVHD altering recovery of in vitro functional responses to the different polyclonal activators. PMID- 2986747 TI - Tac-positive, HTLV-negative, T helper phenotype chronic lymphocytic leukemia cells. AB - In this report we describe an Italian patient with chronic T cell leukemia whose proliferating cells were mature T lymphocytes with a helper phenotype (T helper phenotype chronic lymphocytic leukemia, or Thp-CLL). Unlike other reported cases of Thp-CLL, fresh leukemic cells from this patient were positive with the anti Tac monoclonal antibody, which recognizes the receptor for interleukin-2 (IL-2). Thus, the phenotype of these cells was similar to that expressed by Japanese patients with adult T cell leukemia (ATL) (OKT3+, OKT4+, OKT8-, Tac+). However, the Italian patient had Thp-CLL, not ATL, since his cells, unlike ATL cells, lacked human T cell leukemia virus (HTLV-I)-related DNA sequences. The Tac receptor, which appears to be modulated in vitro by the anti-Tac antibody, was biologically inactive, since the patient's cells did not respond in vitro to IL 2. In addition, they also failed to demonstrate in vitro functional activities. The clinical course was aggressive, as usual, for both Thp-CLL and ATL. Taking advantage of the description of this case, some similarities and differences between Thp-CLL and ATL are discussed, focusing on the importance of screening of HTLV-I in the differential diagnosis. PMID- 2986749 TI - Some biochemical observations on gluconeogenesis from propionate in hepatocytes isolated from normal and biotin-deficients rats. AB - Propionate and pyruvate added to isolated normal and biotin-deficient adult rat hepatocytes increase the production of glucose. This production decreases about 30% on biotin deficiency. Malonate inhibits gluconeogenesis from propionate showing the metabolic transformation of propionyl-CoA via the Krebs cycle. Neither glucagon nor dibutyryl-cyclic AMP significantly stimulate gluconeogenesis. PMID- 2986748 TI - LAV/HTLV III presence in peripheral blood lymphocytes of seropositive young hemophiliacs. AB - Recent studies indicate a high prevalence of seropositivity to the lymphadenopathy-associated virus/human T-lymphotropic virus (type III) among individuals with hemophilia exposed to clotting factor concentrates prepared from large donor pools. The peripheral blood lymphocytes of 19 young seropositive patients with inherited bleeding disorders were examined for the presence of this virus by coculture with phytohemagglutinin-stimulated lymphocytes. Viral isolates were obtained from six of 19 patients. While none of these patients have developed the acquired immunodeficiency syndrome (AIDS) or AIDS-related complex, five of them had lymphadenopathy in two noncontiguous areas, and two showed clinically symptomatic enlarged tonsils and adenoids. Of the 13 patients in whom virus was not demonstrated, five were judged clinically normal and five had mild lymphadenopathy in one anatomical area. These results suggest that as many as 33% of hemophiliacs (six of 19 patients studied), who have circulating antibodies to mature viral proteins, have viral-infected peripheral blood lymphocytes capable of infecting other lymphocytes in vitro. PMID- 2986750 TI - The role of peptidases in cancer of the rectum and sigmoid colon. AB - Increased levels of peptidases are found in some human carcinomas and may be related to invasive potential. We therefore measured the activity of four peptidases in 50 specimens of tumour and normal colonic wall from patients with a rectal or sigmoid carcinoma, and correlated this with the stage, differentiation, fixity of the tumour and presence of venous invasion, determined histologically. Since acute phase reactant proteins (APRP) may inhibit these proteolytic enzymes we have also measured serum levels of two relevant APRPs, alpha 1 acid glycoprotein (AGP) and C-reactive protein (CRP) pre-operatively. Activity of cathepsin B, cathepsin H and collagenase-like peptidase (CLP) was determined fluorimetrically and collagenase photometrically. Significantly elevated activity of cathepsin B, CLP and collagenase was found in tumour compared with normal colonic wall (median values: (nmol (mg protein)-1 min-1) Cat B 0.71 and 0.42 (P less than 0.001), CLP 25.24 and 12.25 (P less than 0.0001) and collagenase 0.49 and 0.31 (P less than 0.001). There was no correlation between the activity of these enzymes expressed as a ratio of tumour/colonic wall, and differentiation or Dukes' stage of the tumour. However, there was significant elevation of activity of cathepsin B in tumours with local spread (n = 13) compared with those with no spread (n = 37) (median values 2.76 and 1.36 respectively (P less than 0.001] and also in tumour with venous invasion (n = 24) compared with tumours without (n = 26) (median values 1.82 and 1.18 respectively (P less than 0.01]. Pre-operative serum levels of CRP were inversely correlated with the activity of CLP and cathepsin H and collagenase in the tumours (rs = 0.332, 0.359 (P less than 0.05) and 0.302 (P = 0.05) respectively). Thus certain peptidases are raised in rectal and sigmoid tumours. Activity of cathepsin B appears related to local tumour invasion. APRP may have a role in inhibiting the activity of these enzymes. These findings may have therapeutic implications. PMID- 2986751 TI - Alpha thalassaemia in British people. AB - Although alpha thalassaemia is rare in north Europeans, it has been identified in British people with no known foreign ancestry. Twelve such patients were studied, of whom eight shared a distinctive molecular defect, which was clearly different from defects seen in subjects of Mediterranean or South East Asian origin. A rare but specific form of alpha thalassaemia is therefore present in the British population. In addition, two patients from families of mixed racial origin were encountered who had a moderately severe form of thalassaemia (HbH disease) due to the inheritance of one form of alpha thalassaemia from the British parent and another type from the foreign parent. This shows the importance of careful genetic counselling of British patients with haematological findings of thalassaemia. PMID- 2986753 TI - Effect of age on response of cattle to vaccination against foot-and-mouth disease. PMID- 2986752 TI - Antibody guided irradiation of brain glioma by arterial infusion of radioactive monoclonal antibody against epidermal growth factor receptor and blood group A antigen. AB - In a patient with recurrent grade IV glioma of the brain resistant to conventional treatment an antibody guided isotopic scan showed uptake by the tumour of a monoclonal antibody (9A) that was developed against epidermal growth factor receptor but cross reacted with blood group A antigen. As a therapeutic attempt antibody labelled with 1665 MBq (45.0 mCi) iodine-131 was delivered to the tumour area by infusion into the internal carotid artery. Computed tomography showed regression of the tumour after treatment, and an appreciable and sustained clinical improvement was noted without any toxicity. Delivery of irradiation guided by monoclonal antibody delivered by arterial infusion of the tumour area may be of clinical value in the treatment of brain gliomas resistant to conventional forms of treatment. PMID- 2986754 TI - Counter-immunoelectrophoresis for rapid diagnosis of sheep pox. PMID- 2986755 TI - The isolation of infectious bursal disease virus from turkeys in England. PMID- 2986756 TI - Comparison of virus neutralizing and precipitating antibodies to infectious bursal disease virus and their effect on susceptibility to challenge. PMID- 2986757 TI - Quantitative evaluation of syncytium formation in cell culture by British isolates of Aujeszky's disease virus. PMID- 2986758 TI - Genome analysis of virulent and attenuated strains of canine parainfluenza virus. PMID- 2986759 TI - Characterization of long-lasting histaminergic inhibition in a beating pacemaker neuron of Onchidium. AB - A single BPSP (excitatory-inhibitory postsynaptic potential) was monosynaptically produced in an identified Onchidium neuron, Be-1, with a beating rhythm upon stimulation of the cardiac nerve. The BPSPs summated to produce an inhibition of long duration (ILD) upon blockage of the beating rhythm after repeated stimulation, so that the BPSPs seemed to be functionally inhibitory. Ten stimuli (1-2 Hz) applied to the cardiac nerve usually evoked an ILD (0.5-1 min) of about 10 mV. The early and middle phases of this ILD reversed near -80 to -85 mV, but the late phase did not reverse at more negative potentials. None of the phases was significantly affected by low Cl or Na solutions or by high Ca solutions. However, by changing the external K, the shift of the reversal potentials for the early and middle phases reached about 65% of that predicted for the K electrode, although the late phase was insensitive to the external K. Intracellular tetraethylammonium (TEA) attenuated the amplitude of the ILD but did not shorten the duration. These suggest that the ILD has another conductance-independent mechanism simultaneously with the increase in K conductance. Several lines of evidence suggested that a ouabain-sensitive Na pump does not contribute to the ILD. Inhibitors of energy supply, 2,4-dinitrophenol sodium salt (DNP) and cyanide, selectively and reversibly reduced the ILD. Simultaneous applications of intracellular TEA and DNP completely abolished the ILD. As for the ionic basis, the histamine-induced inhibitory response in Be-1 was closely related to the ILD. Cimetidine specifically blocked the ILD and histamine-induced inhibitory response, which were mimicked by 2-methylhistamine, but not by dimaprit. It is concluded that the ILD, mediated by some histamine receptor other than the H1 or H2 type, results from an increase in K conductance and a hyperpolarizing ion pump insensitive to ouabain. PMID- 2986760 TI - Estradiol benzoate decreases nigral GABAergic activity in male rats. AB - Repeated doses of estradiol benzoate (10 micrograms/kg, s.c., once a day for 2, 5 or 8 days) to male rats decreased gamma-aminobutyric acid (GABA) content and glutamate decarboxylase (GAD) activity in substantia nigra (SN) but failed to change these parameters in hippocampus, cerebral cortex, cerebellum, lateral septum and olfactory tubercle. In the caudate nucleus, estradiol benzoate decreased GABA concentration but did not modify GAD activity. A decrease in nigral GABA concentration and GAD activity was also observed 24 and 48 but not 3 h after a single injection of estradiol benzoate. These data are consistent with results on GAD activity reported by McGinnis et al. in ovariectomized rats. Kinetic analysis of nigral GAD activity revealed that repeated estradiol benzoate injection reduced the Vmax without affecting the Km of GAD. Estradiol benzoate also reduced the rate of nigral GABA accumulation resulting from local infusion of gabaculine, suggesting that the steroid decreases GABA turnover in male rat SN. Hypophysectomy decreased GABA content and GAD activity in SN and GABA content in striatum. Administration of estradiol benzoate for 8 days to hypophysectomized rats failed to decrease further these parameters. Taken together, these data suggest that estradiol benzoate decreases SN GABAergic activity and that the integrity of the pituitary gland is required for this effect. PMID- 2986761 TI - Effect of GABA and its antagonists, bicuculline and picrotoxin, on nerve cell discharges of the photosensory pineal organ of the frog, Rana esculenta. AB - The effect of gamma-aminobutyric acid (GABA) and its antagonists, bicuculline and picrotoxin, was studied on pineal neurons of the frog, Rana esculenta. The drugs were applied by microiontophoresis while monitoring the spontaneous activity and light-evoked responses of electrophysiologically identified achromatic (luminance) neurons of the pineal organ. Almost all neurons investigated were sensitive to GABA. The inhibitory action was characterized by its rapid onset and its reversibility. The GABA antagonists, bicuculline and picrotoxin, were able to antagonize the inhibitory action of the amino acid. The light-evoked inhibition of the maintained ganglion cell activity interfered with the GABA-induced inhibition, i.e. light reduced the strength of inhibition and shortened the effect of GABA. The investigation suggests a major role of GABAergic mechanisms in the ganglion cell output of pineal neurons. PMID- 2986762 TI - Corticosterone modulation of neurotransmitter receptors in rat hippocampus: a quantitative autoradiographic study. AB - The effect of adrenalectomy (ADX) and corticosterone (CORT) replacement on neurotransmitter receptors was studied in dorsal hippocampus of rat using quantitative autoradiography. ADX for one week causes an increase in [3H]5-HT binding to 5-HT1 receptors which is significant in the CA1 cell field. CORT treatment of ADX rats for 3-5 days results in localized reductions of [3H]5-HT binding including a partial reversal of the increase observed after ADX in CA1. CORT treatment of ADX animals also decreases binding of [3H]QNB to muscarinic receptors in the dorsal hippocampus, with a significant effect in an area designated as subiculum. No influence of CORT was detected on [3H]prazosin binding to alpha 1 adrenergic receptors in dorsal hippocampus. Possible mechanisms for hormone effects on neurotransmitter receptor levels are discussed. PMID- 2986764 TI - Two different mechanisms control inhibition of spike discharge in neurons of cat motor cortex after stimulation of the pyramidal tract. AB - Intracellular recordings were made from neurons of the motor cortex of awake cats while the pyramidal tract (PT) was stimulated at the level of the facial nucleus. In some neurons IPSPs of 35-120 ms peak latency were recorded that diminished in size or reversed with hyperpolarizing current. During these IPSPs a decrease in input resistance reflective of a conductance increase was measured. More often, however, PT stimulation produced IPSPs with comparable latencies that increased in size with hyperpolarizing current. These IPSPs diminished with depolarizing current, and in some instances they appeared to reverse with strong depolarization. During these IPSPs an increased input resistance reflective of a decreased conductance was measured. The results indicate that two different mechanisms control rapid inhibition of spike discharge in neurons of the motor cortex after PT stimulation. PMID- 2986763 TI - The autoradiographic localization of substance P receptors in the rat and bovine spinal cord and the rat and cat spinal trigeminal nucleus pars caudalis and the effects of neonatal capsaicin. AB - Substance P (SP) is a putative neurotransmitter in the central nervous system. In the present report we have used autoradiographic receptor binding techniques to investigate the distribution of SP receptor binding sites in the rat and bovine spinal cord and in the rat and cat spinal trigeminal nucleus pars caudalis. Although some quantitative differences were evident, all species appeared to have a similar distribution of SP receptor binding sites in both the spinal cord and in the spinal trigeminal nucleus pars caudalis. In the spinal cord the heaviest concentration of SP receptors is located in lamina X, while moderate to heavy concentrations were found in laminae I, II and V-IX. Very low concentrations of SP receptors were present in laminae III and IV. Examination of the cat and rat spinal trigeminal nucleus pars caudalis revealed a moderate density of SP receptor binding sites in laminae I and II, very low concentrations in laminae III and IV, and low to moderate concentrations in lamina V. Rats treated neonatally with capsaicin showed a small (11%) but significant (P less than 0.02) increase in the levels of SP receptor binding sites in laminae I and II of the cervical and lumbar spinal cord while in all other laminae the levels remained unchanged. PMID- 2986765 TI - The possible role of 2-hydroxyestradiol in the development of estrogen-induced striatal dopamine receptor hypersensitivity. AB - In the present study, we have confirmed the existence of a biphasic response in striatal dopamine receptor sensitivity following the administration of estradiol benzoate (EB). This biphasic response consists of a hyposensitive phase 24 h after the last injection of EB, followed by a hypersensitive phase 72 h after the last injection of EB. In contrast to this, the administration of 2 hydroxyestradiol (2-OHE2), a catechol metabolite of estrogen, resulted in a striatal dopamine receptor hypersensitivity at both 24 and 72 h after the last injection of 2-OHE2. Studies on the in vivo metabolism of [3H]estradiol to its [3H]catechol metabolites indicated that the administration of piperonyl butoxide (PBO; a microsomal enzyme inhibitor) significantly decreased the level of [3H]catechol metabolites of [3H]estradiol in the striatum and in the medial basal hypothalamus. In addition, PBO administration resulted in about a 7-fold decrease in the ability of estradiol to induce a striatal dopamine receptor hypersensitivity. These data indicate that the biphasic response in striatal dopamine receptor sensitivity following estrogen, may be mediated by separate molecular mechanisms. The association of the hypersensitive phase with pharmacological doses and/or treatment paradigms, the development of a similar hypersensitivity following the administration of the 2-OHE2 metabolite of estrogen and the attenuation of the estrogen-induced striatal dopamine receptor hypersensitivity in PBO pretreated animals all suggest that this striatal dopamine receptor hypersensitivity may be mediated, at least in part, by the catecholestrogens. PMID- 2986766 TI - Acute effects of lead at central synapses in vitro. AB - The acute effects of lead in the rat CNS in vitro were studied on synaptic transmission in the isolated hemisected spinal cord from newborn rats and on the transport of exogenous GABA, acetylcholine and cis-3-aminocyclohexane carboxylic acid (ACHC) from slices of cerebral cortex from adult rats. Lead had quite variable effects on monosynaptic reflexes and synaptic potentials. When it occurred, the depression of synaptic transmission by lead (typically at 18.5 mumol/liters of added lead acetate) was reversible provided exposure times were less than 15 min; furthermore, depression could be antagonised by increasing the external calcium concentration. Lead had no effect on the postsynaptic responses of motoneurons to the putative transmitters L-glutamate, GABA and glycine or to eledoisin-related peptide. The effects of lead on uptake and release of exogenous GABA and ACHC were dependent on the perfusion buffer employed: minimal effects were seen in solutions buffered with either phosphate or carbonate. When Tris HCl was used as buffer, lead inhibited the uptake of GABA and potentiated the spontaneous release of GABA with an EC50 = 50 mumol/liters as added lead acetate. In Tris HCl buffer, lead acetate (100 mumol/liters) produced a two-fold enhancement in the spontaneous release of acetylcholine under conditions where choline and acetylcholine re-uptake was blocked by hemicholinium. The availability of free lead cations in solution is highly dependent on the concentrations of other ions (particularly phosphates) and the pH. Under the appropriate conditions, lead can inhibit CNS synaptic function acutely in a manner consistent with lead competing with calcium ions in transmitter release processes as has been established for acetylcholine release at peripheral synapses. PMID- 2986767 TI - Laminar distribution of alpha 1- and beta 1-adrenoceptors in ferret visual cortex. AB - The distribution of alpha 1- and beta 1-adrenoceptors has been examined in the ferret visual cortex (area 17), with an autoradiographic procedure using iodine 125 labeled hydroxy-iodophenyl-ethylaminomethyl-tetralone (HEAT) and iodocyanopindolol (ICYP), respectively. The density of ligand binding with ICYP, known to have selective affinity for beta-receptors, was heavy over layers I-III, very low over layer IV, and medium over V and VI. In contrast, binding sites for HEAT, a new ligand selective for alpha-receptors, were diffusely distributed, although preferentially concentrated in layer IV and the upper layers. The distinct laminar distribution of these two receptor types may imply two cortical channels for norepinephrine influence; alpha 1-receptors may be preferentially associated with enhancement of excitatory inputs to layer IV; beta 1-receptors, in contrast, with enhancement of inhibitory responses outside layer IV. PMID- 2986768 TI - Modulation of synaptic GABA receptor binding by membrane phospholipids: possible role of active oxygen radicals. AB - Pretreatment of cerebral synaptic membrane preparations with phospholipase (PLase) A2 invariably induced a significant enhancement of [3H]muscimol binding in a dose-dependent manner with a concomitant elevation of the content of total free fatty acids in the membrane. In vitro addition of various free fatty acids exhibited no profound alteration in [3H]muscimol binding, whereas a significant enhancement of the binding was induced by the pretreatment of the membrane with unsaturated free fatty acids such as arachidonic acid and linoleic acid, but not by that with saturated free fatty acids. None of the inhibitors of arachidonic acid metabolism including indomethacin (an inhibitor of cyclo-oxygenase) and nordihydroguaiaretic acid (an inhibitor of lipoxygenase), however, had a significant preventive action on the augmentation of [3H]muscimol binding. On the other hand, various scavengers for superoxide anion radical such as superoxide dismutase, tiron and nitroblue tetrazolium (NBT) not only suppressed the PLase A2 induced enhancement of [3H]muscimol binding, but also diminished the augmentation of the binding due to PLase C and arachidonic acid. It was also found that a remarkable facilitation of the formation of superoxide anion radical was induced by the treatment of synaptic membrane with PLase A2, PLase C and arachidonic acid, all of which exhibited a prominent stimulation of the binding. In addition, treatment of the membrane with xanthine and xanthine oxidase, a superoxide anion radical generating system, resulted in a profound stimulation of the binding. The PLase A2-induced enhancement of the binding was also attenuated by the scavengers for hydrogen peroxide like catalase as well as by those for hydroxyl radical such as dimethylnitrosoaniline, mannitol, methanol and ethanol, but not by those for singlet oxygen radical including alpha-tocopherol and beta-carotene. The present results suggest that membrane phospholipids may play an important role in the modulation of the association of GABA with its relevant receptor through the generation of active oxygen radicals from unsaturated free fatty acids which are yielded by the catalytic action of PLase A2 and/or PLase C. PMID- 2986769 TI - Failure of ES 52, a highly potent enkephalinase inhibitor, to affect nociceptive transmission by rat dorsal horn convergent neurones. AB - The effects of ES 52, a highly potent derivative of the enkephalinase (enkephalin dipeptidylcarboxypeptidase) inhibitor thiorphan, were studied on nociceptive activities of dorsal horn convergent neurones in the anaesthetized rat. Neither the C-fibre component of the responses elicited by supramaximal electrical stimulation of the hindpaw excitatory receptive fields nor diffuse noxious inhibitory controls triggered by immersion of the tail in 46-48 degrees C waterbaths, were affected by ES 52. Thus we conclude that, in our experimental conditions, modulations of the transmission of nociceptive messages at the spinal level are not greatly modified by specifically blocking the degradation of enkephalins. If a major role for enkephalinase (vs aminopeptidase) in the catabolism of enkephalins at the spinal level can be confirmed, then comparison of the present data with our previous results obtained using the opioid antagonist naloxone, might suggest a predominant role for proenkephalin B products (i.e. dynorphins and/or alpha-neo-endorphin) in modulating nociceptive transmission in the spinal cord. PMID- 2986770 TI - Glutamic acid decarboxylase activity decreases in mouse neocortex after lesions of the basal forebrain. AB - Glutamic acid decarboxylase (GAD) activity was measured in the cerebral cortex of animals after acute and chronic lesions to basal forebrain cholinergic nuclei. Such lesions were shown to result in an extensive depletion of cholinergic markers in parietal cerebral cortex. A statistically significant 30% decrease in GAD activity was first detected at 6 weeks postlesion and was still measurable 8 months after the lesion. These results suggest that cholinergic inputs to cortex indirectly or directly influence GABAergic transmission in cortex. PMID- 2986771 TI - Brain somatostatin receptor subpopulation visualized by autoradiography. AB - [125I-Tyr11]somatostatin-14 as well as iodinated D-Tyr1 and Tyr3 derivatives of the cyclic octapeptide somatostatin analog, SMS 201-995 (H-D-Phe-Cys-Phe-D-Trp Lys-Thr-Cys-Thr-ol) have been used as radioligands for somatostatin receptor autoradiography in rat brain. Although the cyclic octapeptide ligands label the majority of the regions labeled with [125I-Tyr11]somatostatin, in the cortex and hippocampus only a subpopulation of somatostatin receptors is labeled. Cyclic octapeptide ligands have improved resolution due to their very low non-specific binding. PMID- 2986772 TI - Stimulation of spontaneous transmitter release by the phorbol ester, 12-O tetradecanoylphorbol-13-acetate, an activator of protein kinase C. AB - Treatment of frog neuromuscular preparations with the protein kinase C activator 12-O-tetradecanoylphorbol-13-acetate (TPA) (100 nM) resulted in a 40% rise in the frequency of miniature endplate potentials (MEPPs) (P less than 0.05). When these experiments were repeated on preparations bathed in saline containing 10 mM K+ (a procedure that raises MEPP frequency by approximately 10-fold due to the elevated level of [Ca2+]i), TPA caused a 70-75% increase in MEPP frequency (P less than 0.02), although the data did not differ significantly from those recorded in standard frog saline due to the variability between preparations. It is concluded that transmitter release mechanisms may be modulated by protein kinase C and that such modulation probably acts synergistically with [Ca2+]i. PMID- 2986773 TI - Excitatory and inhibitory effects of dopamine on synaptic transmission in the rat olfactory cortex slice. AB - A study has been undertaken of the effects of dopamine on excitatory transmission at the lateral olfactory tract (LOT)-superficial pyramidal cell synapse of the rat olfactory cortex slice by measuring the effects of bath-applied dopamine on the amplitudes and latencies of the surface field potentials evoked on submaximal LOT stimulation in a total of 32 preparations. In 7 (22%) slices, dopamine had no detectable effects on transmission. In the remaining preparations, dopamine (1 250 microM) depressed transmission in a concentration-dependent manner. This action was unaffected by nadolol (10 microM), phentolamine (10 microM) and picrotoxin (25 microM) but was antagonized by chlorpromazine (10 microM) and trifluoperazine (0.2 and 0.5 microM) and mimicked by bromocriptine (0.01-5 microM) and apomorphine (0.25-25 microM). Investigation of the effects of dopamine on stimulus input-evoked potential output relationships indicated that the inhibitory effect of dopamine on transmission was mediated by a reduction in pyramidal cell excitability. In 6 slices (24% of those sensitive to dopamine) low dopamine concentrations (0.1-1 microM) facilitated transmission at the LOT superficial pyramidal cell synapse. This excitatory effect was antagonized by nadolol and phentolamine (10 microM) and also by 100 microM 2-amino-5 phosphonovalerate (an antagonist of excitatory amino acid receptors of the N methyl-D-aspartate type) but was unaffected by chlorpromazine (10 microM) and trifluoperazine (0.2 and 0.5 microM). By a comparison with the effects of noradrenaline on transmission, it is concluded that the excitatory effects of dopamine are mediated either indirectly by the release of noradrenaline or by a direct interaction of dopamine with adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986774 TI - Dopaminergic mechanisms in subthalamic nucleus of rat: analysis using horseradish peroxidase and microiontophoresis. AB - Afferent connections to the subthalamic nucleus (STN) were studied by microiontophoretically injecting horseradish peroxidase (HRP) into the STN and studying its retrograde transport. Remotely labelled neurons were frequently observed in both the globus pallidus and the pars compacta region of substantia nigra. In addition, individually labelled neurons were occasionally found in other brain regions. The sensitivity of neurons in the STN to dopamine (DA) was studied by applying DA to neurons in the STN by microiontophoresis. Three patterns of response to DA were observed. The most frequent response, observed in 46% of the STN neurons studied, was a decrease in the discharge frequency. In 15% of the neurons there was an increased frequency of firing. Eleven percent of the neurons exhibited a mixed response consisting of an initial depression of discharge rate followed by a delayed increase. The responses of STN neurons to DA were not antagonized by iontophoretically applied haloperidol. In neurons whose firing frequency was decreased by DA, the iontophoretic application of apomorphine and norepinephrine also decreased discharge rate. The observations of HRP-labelled neurons in the pars compacta region of substantia nigra following injection of HRP into the STN together with the DA responsiveness of STN neurons suggest the possibility of a dopaminergic nigro-subthalamic pathway. PMID- 2986775 TI - Opposing effects of striatonigral feedback pathways on midbrain dopamine cell activity. AB - The existence of a striatonigral GABAergic pathway has been well established both anatomically and biochemically. During intracellular recording from identified DA neurons in vivo, stimulation of the striatum (100 microA, 50 microseconds pulses) elicits an inhibitory postsynaptic potential (IPSP) and a rebound depolarization. The IPSP is a short latency (1.8-2.2 ms) conductance increase to chloride, since: the reversal potential is near the chloride reversal potential reported for other cells (-68 mV); intracellular chloride injection progressively reverses the IPSP into a depolarization with a similar time course; and the response of DA cells to systemic injection of the chloride channel blocker, picrotoxin, also exhibits a similar reversal potential. In contrast, during extracellular recording, stimulation of the striatum at low levels of intensity (e.g. 20 microA at 10 Hz) increases the firing rate of DA cells. Stimulation of the striatum will, in addition, elicit IPSPs in a subclass of substantia nigra zona reticulata neurons at the same latency as the IPSPs triggered in DA cells. These IPSPs also reverse with intracellular chloride injection. However, their amplitude is larger and their duration longer than observed in DA cells, and there is no depolarizing rebound. The late component of the IPSP in the zona reticulata neurons corresponds temporally to the rebound depolarization seen in DA cells in response to striatal stimulation. In addition, when recorded extracellularly, striatal stimulation will inhibit the firing of this class of zona reticulata interneurons at the same stimulation parameters that will excite DA cells. These data suggest that striatal cells may send branched fast-conducting GABAergic projections to zona reticulata cells and DA cells. Furthermore, low levels of striatal stimulation can excite DA cells by preferentially inhibiting interneurons in the zona reticulata which are more sensitive to the inhibitory effects of GABA than are DA neurons. PMID- 2986776 TI - Perforant pathway-evoked long-term potentiation of CA1 neurons in the hippocampal slice preparation. AB - Previously, we have presented electrophysiological evidence reaffirming the existence of a controversial hippocampal pathway. These fibers are part of the perforant pathway and terminate directly on the CA1 cells. We now report that, in the hippocampal slice preparation, tetanic stimulation of the perforant pathway produces long-term potentiation (LTP) of CA1 cell responses. LTP of population spikes varied from 150% to 500%. The results were of interest because these axons synapse at distal sites on the apical dendrite. This location is usually thought to be a difficult site to evoke action potentials. PMID- 2986777 TI - Effects of inhibitors of Na+,K+-ATPase on the membrane potentials and neurotransmitter efflux in rat brain slices. AB - The potassium potential EK, of rat brain slices was estimated by determining the uptake of 86Rb+. The ERb was the same for slices prepared from five rostral brain regions, the average value being 66.4 mV. The ERb values in the presence of 20 microM ouabain were only slightly lower than the resting values; increasing concentrations of ouabain above 20 microM resulted in a graded depolarization in all five brain regions. High concentrations (1 mM) of two other inhibitors of Na+,K+-ATPase, dihydro-ouabain and strophanthidin, produced no more depolarization than did 20 microM ouabain. Competitive binding studies indicated that the differential effects were due to the relative binding to brain slices. Erythrosin B, an inhibitor of Na+,K+-ATPase, had no measurable effect on ERb. Intermediate concentrations of the Na+/H+ ionophore monensin slightly hyperpolarized striatal slices, whereas the same monensin concentrations plus 20 microM ouabain, 1 mM strophanthidin or 70 microM erythrosin B resulted in marked depolarization. Measurement of the membrane potential via uptake of methyltriphenylphosphonium cation indicated that ERb was indeed a valid estimation of the membrane potential. EK was measured directly by monitoring 42K+ uptake in striatal slices and was found to be essentially identical to ERb. Uptake of 22Na+ was consistent with the values for ERb or EK. Several conditions that resulted in little or no measurable depolarization of striatal slices did induce efflux of exogenously loaded GABA and dopamine; these conditions included 20 microM ouabain, 1 mM dihydro-ouabain or strophanthidin, and 70 microM erythrosin B. Neurotransmitter efflux in the absence of general cell depolarization was not accompanied by altered rates of respiration or decreased ATP levels. PMID- 2986778 TI - Muscarinic acetylcholine receptors are localized on striatal cyclic GMP containing neurons. AB - The localization of muscarinic acetylcholine receptors has been determined using in vitro binding of radiolabeled quinuclidinyl benzilate ([3H]QNB), a specific reversible muscarinic receptor antagonist. All cyclic GMP-immunoreactive neurons in the rat striatum show clustering of [3H]QNB silver grains overlying their somata following autoradiographic analysis. The autoradiography of total binding of silver grains over the cyclic GMP-containing neurons was approximately 800 times as dense as the surrounding neuropil localization of radioligand binding sites. Incubation of striatal tissue slices in the presence of micromolar atropine, to determine non-specific binding of [3H]QNB, decreased the autoradiographic silver grain density of the neuropil about 2.5 times, and lessened the number of receptor sites detectable on cyclic GMP-positive neurons at least 5-fold. Biochemical examination of [3H]QNB binding on tissue sections demonstrated that the ligand binding is saturable and dependent on section thickness for the muscarine receptor subtype of acetylcholine. PMID- 2986779 TI - Dendrodendritic synapses in the cat reticularis thalami nucleus: a structural basis for thalamic spindle synchronization. AB - Dendrodendritic synapses have been found between the dendrites of reticularis thalami neurons. These synapses were symmetric and the presynaptic element contained pleomorphic vesicles. A few cases of reciprocal dendrodendritic synapses were also observed. Given the morphological features of the synapses and the well-established GABAergic nature of reticularis neurons it is concluded that reticularis cell dendrites form a local inhibitory network. The functional implications of this type of organization for the synchronization of spindle oscillations are discussed and a new hypothesis is proposed. PMID- 2986780 TI - Dopamine receptors regulate Met-enkephalin content in pituitary. AB - Met-enkephalin-like immunoreactive material (ME-LI) in the neurointermediate lobe of rat pituitary was dramatically altered by the long-term administration of drugs altering dopamine receptor activity. Treatment with the dopamine receptor antagonist haloperidol resulted in significant increases of ME-LI and treatment with the dopamine receptor agonist bromocriptine resulted in significant decreases of ME-LI in the neurointermediate pituitary. The treatments had considerably less effect on ME-LI in the anterior lobe of the pituitary. These results suggest that dopamine receptor activation or inhibition influences the content of ME-LI in the rat pituitary gland. PMID- 2986781 TI - Association of the mesencephalic locomotor region with locomotor activity induced by injections of amphetamine into the nucleus accumbens. AB - Injections of amphetamine into the nucleus accumbens increased locomotor activity of rats. Subsequent injections of procaine into the midbrain, in the region of the pedunculopontine nucleus, significantly reduced the amphetamine-induced locomotor activity. Control experiments showed that procaine injections into the contralateral pedunculopontine nucleus had little or no effect, as well as ipsilateral injections dorsal and ventral to the pedunculopontine nucleus. These findings suggest that release of dopamine from amphetamine injections into the accumbens gives rise to ipsilateral descending influences on the region of the pedunculopontine nucleus, a major component of the mesencephalic locomotor region. Descending influences from the nucleus accumbens to mesencephalic locomotor region may serve as a link for limbic-motor integration in behavioral response initiation. PMID- 2986783 TI - An alpha 2 receptor mediates the selective inhibition by noradrenaline of nociceptive responses of identified dorsal horn neurones. AB - Extracellular recordings were made of 59 neurones with long, ascending projections (spinocervical tract (SCT) and dorsal column postsynaptic (DCPS) neurones) in the lumbar dorsal horn of anaesthetized and paralyzed cats. All showed prominent excitatory responses to innocuous stimuli, applied to their cutaneous receptive fields on the ipsilateral hindlimb. The majority of the population investigated (83%) was multireceptive, being activated by noxious as well as innocuous cutaneous stimuli. Drug effects were examined on a regular cycle of responses to these cutaneous stimuli and also to DL-homocysteic acid (DLH). In 49 multireceptive SCT and DCPS neurones, ionophoretically-applied L noradrenaline (NA) produced a potent selective inhibition of the nociceptive responses (to heat or pinch) in 40 out of 44 SCT and 3 out of 5 DCPS neurones, with no statistically significant change in the responses to innocuous brush or DLH, or in spontaneous activity. NA had no effect on the majority of cells (8 out of 11) that responded only to innocuous stimuli. In 19 SCT neurones that showed NA-selectivity, the alpha 2-selective agonists clonidine (in 12 out of 15) and metaraminol (in 2 out of 3) mimicked this selective effect, whereas, the alpha 1 agonist, phenylephrine and the beta agonist, isoprenaline did not. Furthermore, the alpha 2 antagonists, yohimbine and idazoxan (RX781094), either reversed or reduced the potency of the NA-elicited inhibition of nociceptive responses in all 7 SCT neurones tested. These results are discussed in relation to other evidence for spinal antinociceptive effects of noradrenergic systems acting at a spinal level and the possible involvement of an alpha 2 receptor in such effects. PMID- 2986782 TI - The hypothalamohypophyseal system in vitro: electrophysiology of the pars intermedia and evidence for both excitatory and inhibitory inputs. AB - The purpose of this study was to better assess the function of catecholamine containing nerve terminals in the pituitary pars intermedia lobe. Hypothalamohypophyseal explants, which included the intact mediobasal hypothalamus (MBH), median eminence, infundibular stalk and the neurointermediate lobe, were obtained from 2-3-week-old male and female albino rats. The tissue was placed in a perfusion chamber and maintained under physiological conditions for up to 12 h. A set of bipolar stimulating electrodes was positioned on the surface of the median eminence, infundibular stalk or the rostroventral arcuate nucleus of the MBH. A microelectrode recorded electrical activity in the pars intermedia gland. Two types of spontaneous action potentials were found; fast 2-4 ms duration neural fiber type spikes and slower 7-10 ms duration spikes probably derived from non-neural endocrine cells. Single-pulse electrical stimulation at all 3 sites evoked both kinds of potentials, while trains of stimuli (0.1-20 Hz) decreased or completely inhibited the basal firing rate of the slower ones. Application of the neuroleptic. L-sulpiride (0.01, 0.1 or 1.0 mumol), to the perfusion medium increased the spontaneous endocrine cell activity and blocked the stimulus-induced inhibition in the explants but had no effect on the activity in isolated pituitaries. Dopamine (0.1 mumol), which is known to inhibit the secretion of pro-opiomelanocortin peptides, reversibly suppressed the spontaneous endocrine cell potentials. These observations support a hypothesis for the presence of a functional tuberohypophyseal dopamine inhibitory system and a possible, but as yet unidentifiable, excitatory system in the pars intermedia. Thus, hypothalamohypophyseal explants can be used to elucidate specific information on this type of neuroendocrine axis. PMID- 2986784 TI - Localization of renin (EC 3.4.23) and converting enzyme (EC 3.4.15.1) in nerve endings of rat brain. AB - Synaptosomes and lysosomes of rat brain were separated by differential centrifugation and a two-step gradient centrifugation with colloidal silica-gel (Percoll). The organelles were identified by the measurement of established marker-enzymes and by electronmicroscopy. Renin activity, measured by radioimmunoassay for angiotensin I (ANG I), was localized in the synaptosomes and cathepsin D-activity was found in the lysosomal fraction. Converting-enzyme activity was present in the renin-containing synaptosomes. Part of the brain renin activity could be activated by pre-incubation with trypsin. Affinity chromatography of an organelle-enriched brain fraction was carried out using a caseinyl-sepharose column and resulted in the separation of renin from cathepsin D activity; the renin peak was inhibited by antibodies raised against rat kidney renin. We conclude, that the formation of ANG I and its activation to angiotensin II (ANG II) by converting enzyme is possible in synaptosomes. This adds further evidence to an intraneuronal synthesis of ANG I and ANG II in the brain and is in support of previous results demonstrating an intraneuronal localization of the components of the brain renin-angiotensin system. PMID- 2986785 TI - Effects of synaptic antagonists on perforant path paired-pulse plasticity: differentiation of pre- and postsynaptic antagonism. AB - The effects of different synaptic antagonists on paired-pulse plasticity of medial perforant path responses were studied in rat hippocampal slices. Baclofen reduces the response to activation of the perforant path, but does not have the same net effect on the first and second responses to paired stimulation: baclofen lessens the percent paired-pulse depression of medial perforant path responses. Furthermore, at doses that reduced the control medial perforant path response by half, paired-pulse plasticity changed from paired-pulse depression to paired pulse potentiation. A similar effect on medial perforant path paired-pulse plasticity is produced by decreasing extracellular calcium concentration. Kynurenic acid reduces the first and second responses to paired stimulation proportionately the same, and, therefore, has no effect on the percent paired pulse depression. These results suggest that baclofen acts presynaptically to reduce the synaptic response, whereas kynurenate acts postsynaptically. Adenosine was also found to be a potent antagonist of medial perforant path responses, with effects on paired-pulse plasticity similar to baclofen: a new synaptic antagonist, N-p-chlorobenzoyl-piperazine-2,3-dicarboxylate, was found to have effects like kynurenate, suggesting that it is also a postsynaptic receptor blocker. PMID- 2986786 TI - Prenatal exposure to diazepam results in enduring reductions in brain receptors and deep slow wave sleep. AB - After prenatal exposure to diazepam (Valium), mature rats at 4 months of age displayed slow wave sleep (SWS) electroencephalographic patterns indicating impaired synchronization and SWS mechanisms. These animals spent a much greater portion of their SWS in the lighter SWS I, as compared to the control group which showed a predominance of the deeper SWS II. At one year of age, the diazepam exposed rats had much fewer diazepam-specific binding sites in the thalamus than the vehicle-exposed controls. These results provide first evidence for a physiological role for benzodiazepine receptors by showing that prenatal exposure to diazepam has an enduring and detrimental effect on their ontogenesis and sleep mechanisms. PMID- 2986787 TI - Cyclohexyladenosine binding in rat striatum. AB - N6-Cyclohexyl-[3H]adenosine [( 3H]CHA) binds specifically to rat brain membranes prepared from the caudate-putamen complex with a Kd value of 2.50 +/- 0.39 nM and Bmax of 458 +/- 51 fmol/mg protein. Lesioning the nigrostriatal dopaminergic pathway using 6-hydroxydopamine failed to alter [3H]CHA binding characteristics. Intrastriatal kainate lesions reduced the binding capacity of [3H]CHA by 28% though this was not statistically significant (0.1 less than P greater than 0.05). In kainate-lesioned striata, however, 2-deoxyglucose uptake was reduced by only 39%. PMID- 2986789 TI - Electrophysiological studies on the postnatal development of intracerebellar nuclei neurons in rat cerebellar slices maintained in vitro. I. Postsynaptic potentials. AB - The development of the synaptic responses of intracerebellar nuclei neurons was studied in the rat by the use of thick sagittal cerebellar slices maintained in vitro. It has been shown that functional excitatory synapses are present on these neurons from birth, probably due to climbing and/or mossy fiber collaterals; functional inhibitory synapses, due to monosynaptic projections of Purkinje cell axons onto intracerebellar nuclei, are present as early as postnatal day 2; and a more complex pattern of synaptic responses, including short latency excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs), longer latency IPSPs, and late depolarizing responses, can be elicited in nuclear neurons as early as postnatal day 3, indicating an early development of some complete functional cerebellar circuits involving the intracerebellar nuclei. PMID- 2986788 TI - Differentiation characteristics of human neuroblastoma cells in the presence of growth modulators and antimitotic drugs. AB - Morphological characteristics of undifferentiated and differentiated human neuroblastoma cells were studied. Monolayer cultures of a human neuroblastoma, IMR-32 clone, were grown in Eagle's minimum essential medium with fetal calf serum in tissue culture dishes with polystyrene film liners. After 48 h, cultures were treated with either mitomycin C and 5-bromodeoxyuridine or prostaglandin E1 (PGE1) and dibutyryl adenosine 3',5'-cyclophosphate (cAMP). A third dish was untreated to study as an undifferentiated control. Three days later, all cultures were processed for acetylcholinesterase staining, scanning and transmission electron microscopy and high performance liquid chromatography. Treatment with mitomycin/5-bromodeoxyuridine and PGE1/cAMP inhibited growth as seen by the growth curves and caused morphological differentiation as seen by the extension of long neurites. The treated cells showed increased acetylcholinesterase staining compared to the controls. With the scanning electron microscope, the differentiated cells showed long neurites, processes with beaded varicosities and growth cones. By transmission electron microscopy, these cells contained a large number of neurosecretory granules in their cytoplasm and neurites. Specialized cell contacts were also observed between the treated cells. This is the first study demonstrating that both the treated and control cells of IMR-32 clone contain large quantities of serotonin and comparatively small amounts of norepinephrine and dopamine. PMID- 2986790 TI - Development of serotonergic and adrenergic receptors in the rat spinal cord: effects of neonatal chemical lesions and hyperthyroidism. AB - The sympathetic preganglionic neurons in the spinal cord receive dense serotonergic (5-HT) and catecholaminergic (CA) afferent inputs from the descending supraspinal pathways. In the rat spinal cord, the levels of these biogenic amines and their receptors are low at birth, but undergo rapid ontogenetic increases in the ensuing 2-3 postnatal weeks until the adult levels are reached. In many systems it has been shown that denervation of presynaptic neurons leads to an up-regulation of the number of postsynaptic receptors. To determine whether the 5-HT and CA receptors in the developing spinal cord are also subject to such transsynaptic regulation, we examined the ontogeny of serotonergic receptors and alpha- and beta-adrenergic receptors in thoracolumbar spinal cord of rats given neurotoxins which destroy serotonergic (5,7 dihydroxytryptamine (5,7-DHT)) or noradrenergic (6-hydroxydopamine (6-OHDA)) nerve terminals. Intracisternal administration of 5,7-DHT or 6-OHDA at 1 and 6 days of age prevented, respectively, the development of 5-HT and CA levels in the spinal cord. Rats lesioned with 5,7-DHT displayed a marked elevation of 5-HT receptors with a binding of 50% greater than controls at 1 week and a continuing increase to twice normal by 4 weeks. A similar pattern of up-regulation was also detected with the alpha-adrenergic receptor, as rats lesioned with 6-OHDA exhibited persistent increases in receptor concentration. However, in these same animals ontogeny of the beta-adrenergic receptor in the spinal cord remained virtually unaffected by the chemical lesion. In several other parts of the nervous system, it has been demonstrated that the beta-adrenergic sensitivity can be modulated by hormonal signals, particularly that of the thyroid hormones. This phenomenon was examined in the spinal cord and in confirmation with previous studies neonatal treatment of triiodothyronine (0.1 mg/kg, s.c. daily) was capable of evoking persistent increases in beta-adrenergic receptor binding. These results suggest that: (a) development of the postjunctional serotonergic and alpha-adrenergic receptors in the rat spinal cord can occur in the absence of the prejunctional nerve terminals and are subject to transsynaptic modulation; (b) beta-adrenergic receptors in the spinal cord also can develop after prejunctional lesions but are regulated by hormonal rather than neuronal factors. PMID- 2986792 TI - Immunological study of the rat hypothalamic ventromedial nucleus. AB - Specific anti-sera were prepared by injecting the homogenates of rat ventromedial nucleus (VMH) and caudate nucleus (Cd) into the rabbit. Anti-VMH serum, after absorption of common components in rat normal serum and anti-Cd serum, reacted specifically to the rat VMH with only one precipitation line. Anti-VMH serum was successfully applied to 80 VMH neurons by electrophoresis through 5 barreled micropipettes. Fresh anti-VMH serum caused an irreversible response (increase and subsequent sudden cessation of firing) in 8 of the 25 VMH neurons tested. Anti VMH serum reversibly inhibited 32 of 80 VMH neurons and 13 of these were also tested with glucose. Discharge rates of 12 of the 13 neurons increased by glucose. Most of these neurons were not affected by anti-Cd serum or normal rabbit serum. Results of these immunological and electrophysiological studies suggest the existence of specific membrane receptor binding sites on the glucoreceptor neurons in the VMH. These sites afford one route for producing the excitatory effect that glucose has on VMH neurons. PMID- 2986791 TI - Extranuclear axon collaterals of paraventricular neurons in the rat hypothalamus: intracellular staining, immunocytochemistry and electrophysiology. AB - Recent studies have suggested that some paraventricular nucleus (PVN) neurons projected to more than one target and, thereby, perhaps coordinate some aspects of seemingly diverse functions. We have systematically investigated the existence, location, hormonal contents and functional integrity of some axon collaterals arising from PVN neurons. This was done using intracellular injections of the fluorescent dye, Lucifer Yellow, extracellular ejections of horseradish peroxidase (HRP), immunocytochemistry with antisera directed against vasopressin (VP) and oxytocin (OX) and electrophysiological analysis of synaptic activation of perifornical neurons in response to electrical stimulation of the PVN in hypothalamic slices. Each of the three morphological techniques revealed clear axon collaterals, arising in the lateral hypothalamus and generally ventrolateral to the PVN. Most branching axons appeared to have a small number of branch points, and many collaterals appeared to terminate near their parent axon. Electrical stimulation of the PVN was found to activate synaptically perifornical neurons located in the areas where the other methods revealed collaterals. Stimulation outside of the nucleus was ineffective unless current intensities were increased 10-30-fold over those applied to the PVN. We conclude that many PVN neurons, at least some of these containing OX and other VP, give rise to axons that branch in the perifornical and more ventral lateral hypothalamus, and that some of their collaterals probably terminate on neurons close to the PVN. PMID- 2986794 TI - A simple perfusion system for brain slice electrophysiology. AB - A simple perfusion system for brain slice experiments was designed to improve stability of intracellular recordings and to enable rapid application of drugs. This system can easily be installed in most conventional apparatuses for electrophysiological experiments, with minimal and inexpensive modifications. PMID- 2986793 TI - Heterogeneity of brain benzodiazepine receptors: effects of physiological conditions. AB - A number of investigators have shown compelling evidence for multiplicity of benzodiazepine (BDZ) receptors. The present study addresses the query of BDZ receptor heterogeneity, in vitro, with respect to temperature. In competition studies involving rat cerebellar tissue, CL 218,872 produced Hill slopes near unity at both 0 degree C and 37 degrees C. In contrast, similar experiments utilizing cortical tissue from rats and mice produced Hill slopes of 0.69 and 0.66 at 0 degree C and 37 degrees C respectively. 3H-Flunitrazepam-photoaffinity labeling of cortical and cerebellar membranes was conducted at 0 degree C and 37 degrees C. SDS-PAGE fluorographic analyses of photolysed 3H-flunitrazepam (3H Flu) revealed one intensely labeled 51K band in the cerebellum at both temperatures, which was specifically chased by diazepam. Similar experiments conducted in cortical tissue revealed photoaffinity labeling of at least three distinct macromolecules, one intense 51K and two less intense 55K and 59K bands. Labeling of each of these bands was chased specifically by diazepam. These data, taken together, indicate the existence of regional BDZ receptor heterogeneity under physiological conditions. PMID- 2986795 TI - Morphine and specific changes in the sensitivity of noradrenergic receptors within the "limbic" part of the feline caudate nucleus: a behaviour study. AB - The present study describes the behaviour effects of intracerebral injections of the noradrenergic (NE) agonist oxymetazoline and the NE antagonist phentolamine into the "limbic" part of the caudate nucleus of cats primed 24 hr earlier and/or treated acutely with morphine (5 mg/kg, IP). Drug-induced changes in the morphine specific behaviour served as dependent variables. Experiments were performed during two different periods of the year, each of them marked by a characteristic sensitivity of alpha-like NE receptors to NE agents, viz. the so-called NE "antagonist" period during which the NE receptors were sensitive to the NE antagonist phentolamine and the so-called NE "agonist" period during which the NE receptors were sensitive to NE and the NE agonist oxymetazoline. The present study demonstrates that morphine reversed the initial sensitivity to oxymetazoline respectively insensitivity to phentolamine in animals tested in the NE "agonist" period. In animals tested in the NE "antagonist" period morphine did not reverse the initial insensitivity to oxymetazoline resp. sensitivity to phentolamine. Furthermore, evidence is provided that the initial sensitivity to NE agents did not conspicuously determine the animal's response to the acute administration of morphine. The data are discussed in view of the concept that the firing rate of NE fibres determines the actual sensitivity of presynaptic and postsynaptic NE receptors to NE agonists and antagonists. PMID- 2986796 TI - Retroviruses and human cancer: evaluation of T-lymphocyte transformation by human T-cell leukemia-lymphoma virus. PMID- 2986797 TI - SOS response in mammalian cells. PMID- 2986798 TI - [Microautoradiographic method permitting the study of migration of 99m-technetium labelled leukocytes]. AB - Cell migration studies are very important in inflammatory phenomena. Methods currently used are not quantitative and have been subject to much controversy. Homogeneity of 99mTc leukocyte labelling was verified by a microautoradiographic method (MAR), which was performed in our laboratory. This method was used in cell migration studies to verify if the migrating cells were indeed the labelled cells. PMID- 2986799 TI - [Inhibitory effect of sodium butyrate on the in vitro growth of Herpes virus hominis, type I]. AB - The Herpes virus hominis growth is studied in Vero cells pretreated for 48 hrs with sodium butyrate 24 hour later. Total virus yield is at least ten times less in treated than in infected control cells. Butyrate has no direct effect on Herpes virus survival. However plaque numbers in pretreated cells are reduced by about 60 per 100. PMID- 2986800 TI - [Specific receptors for triiodothyronine in nuclei isolated from normal human polynuclear neutrophils]. AB - In normal subjects, nuclei isolated and purified from circulating granulocytes bound 125I-T3. Binding was reversible and inhibited by unlabelled hormone. Scatchard plots showed a single class of high affinity sites (Kd: approximately 1,5 nM) with a high maximal binding capacity (MBC: approximately 400 fmol of T3 bound/100 micrograms of DNA). Structural analogs partially competed with 125I-T3 binding. These data suggest that human normal polymorphonuclear neutrophils possess specific nuclear receptors for triiodothyronine. PMID- 2986802 TI - [Functional and specific vasoactive intestinal peptide receptors in enterocytes isolated from fetal or adult rats]. AB - Functional and specific VIP receptors (relative potencies: VIP greater than PHI greater than secretin) have been characterized (cAMP generation) in enterocytes isolated from rat fetuses at 19 days gestation, after incubation at 37 degrees C in the presence of IBMX. In fetuses, VIP is about 6 times more potent (EC50 = 2.5 X 10(-10) M) than in adults (EC50 = 15 X 10(-10) M). This difference is not observed for PGE2, and is not related to cAMP-PDE activities (Km, Vmax). It is suggested that VIP may regulate the differentiation and function of enterocytes during the fetal life in rats. PMID- 2986801 TI - [Presence of a peripheral type benzodiazepine binding site on the macrophage; its possible role in immunomodulation]. AB - Saturable binding site for 3H-flunitrazepam (KD = 43 +/- 7 nM, Bmax = 391 +/- 58 fmoles/cell, i.e. 250,000 sites/cell) is characterized on Mouse peritoneal inflammatory macrophages. The affinity for different ligands (PK 11195 greater than Ro 5-4864 greater than diazepam greater than flunitrazepam greater than clonazepam greater than Ro 15-1788) shows that this site is of peripheral type. In vivo the humoral response in Mice to Sheep red blood cells was stimulated by administration of 1 mg/kg of PK 11195 (+85%), Ro 5-4864 (+80%) and diazepam (+58%). Clonazepam and Ro 15-1788 are devoid of activity. This suggests that molecules which show affinity for the "peripheral type" benzodiazepine binding site might modulate the immune response. PMID- 2986804 TI - Evaluation of possible mechanisms of phorbol ester stimulation of phosphatidylcholine synthesis in HeLa cells. AB - Phorbol esters, including 12-O-tetradecanoylphorbol-13-acetate (TPA) and 12,13 dibutyrylphorbol acetate, markedly stimulate the synthesis of phosphatidylcholine (PC) and the activity of CTP:phosphocholine cytidylyltransferase (CT) in cultured HeLa cells. Two possible mechanisms whereby the phorbol esters stimulated PC biosynthesis were investigated. One consideration was that phorbol esters may induce the release of fatty acyl chains from endogenous complex lipids; increased fatty acids or fatty acyl-CoAs could cause the translocation of CT from cytosol to microsomes and thereby increase the activity of the rate-limiting enzyme in PC synthesis. In HeLa cells prelabeled with [3H]oleate or [3H]arachidonate, phorbol ester treatment increased the redistribution of arachidonate in phospholipids and neutral lipids and release of label to the medium, but there was little effect on the cellular fatty acid pools with either of the labeled fatty acids or of the phorbol esters. A second possibility was that protein kinase C (PKC), a receptor for phorbol esters, might be involved in activation of CT activity. TPA stimulated the phosphorylation of cytosolic proteins of HeLa cells more than twofold during a 10- or 60-min incubation with 32Pi. However, an approximate sixfold purified preparation of PKC from rat brain did not stimulate the activity of partially purified (12- to 15-fold) CT; a slight inhibition, dependent on ATP but independent of Ca2+ and diolein, was observed. Our results suggest that intracellular release of free fatty acids or direct phosphorylation of CT by PKC probably do not account for the observed levels of stimulation by phorbol esters. Other possible mechanisms are discussed. PMID- 2986803 TI - 1 alpha,25-dihydroxyvitamin D3 receptors and their action in embryonic chick chondrocytes. AB - The role of vitamin D in the maturation of epiphyseal chondrocytes was investigated in the developing chick embryo. Cartilage tissues were divided into two parts: resting cartilage and growth cartilage. A cytosol component to which 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) is specifically bound first appeared in the growth cartilage on day 15, rapidly increased, and attained a maximum on day 19. The calcium content of the growth cartilage also began to increase on day 15 and continued to increase in parallel with the 1 alpha,25(OH)2D3 receptor levels. Glycosaminoglycan (GAG) synthesis by the growth cartilage cells increased from day 11-17 and rapidly declined thereafter reciprocally with the increase in calcium and receptor levels. In the resting cartilage, no cytosol receptor for 1 alpha,25(OH)2D3 was detected up to hatching time. The calcium content and GAG synthesis in the resting cartilage were very low and did not change appreciably throughout development. No receptor-like macromolecule for 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) was recognized in either the resting or growth cartilage. 1 alpha,25(OH)2D3 added to the culture of chondrocytes from the epiphyseal growth cartilage inhibited GAG synthesis and stimulated its release from the cell layer into the medium in a dose-dependent manner. These in vitro effects of 1 alpha,25(OH)2D3 were not observed in chondrocytes obtained from 13-day-old growth cartilage and 19-day-old resting cartilage. 25-Hydroxyvitamin D3 and 24R,25(OH)2D3 had no effect on chondrocytes in any of the preparations. These results suggest that 1 alpha, 25 (OH)2D3 is directly involved in the maturation of chondrocytes and possibly in the calcification of growth cartilage. PMID- 2986805 TI - Symposium on outpatient anorectal procedures. Alternatives to surgical hemorrhoidectomy. AB - Hemorrhoids are an extremely common affliction. The prevalence ranges from 1 in 25 to 1 in 30 individuals. There was a 20% decrease in the number of hemorrhoidectomies performed between 1978 and 1982. Alternatives to formal hemorrhoidectomy are injection sclerotherapy and rubber-band ligation. High-fibre diets and bulk laxatives are effective in relieving the symptoms of hemorrhoids, easing defecation and regulating bowel habit and can be used liberally. Bleeding hemorrhoids can be treated by diet alone, or by injection or rubber-band ligation. Prolapsing hemorrhoids are best treated by rubber-band ligation. The relatively few patients in whom these more conservative measures fail can be considered for hemorrhoidectomy. PMID- 2986806 TI - Breast cancer risk factors: can we select women for prophylactic mastectomy? AB - Prophylactic procedures to prevent disease in women at high risk for breast cancer should be considered only when the benefits of prophylaxis outweigh its costs. The rational prophylactic use of mastectomy in these circumstances must await answers to several questions. First, accurate estimates of the true risk associated with these lesions must be obtained, as current estimates are imprecise and potentially biased. Second, large ongoing trials of breast cancer screening and treatment may provide effective methods of distinguishing those women at exceptionally high risk in whom such procedures may be warranted. Finally, the question of conservative surgery as an acceptable treatment for breast cancer must be settled; otherwise, the prophylactic procedure may be more invasive and disfiguring than the treatment of the disease it seeks to prevent. PMID- 2986807 TI - Definition of precancerous and high-risk mammary lesions. AB - Intraepithelial (noninvasive) neoplasia of breast, the most potent precursor of invasive carcinoma, comprises two types: lobular carcinoma in situ and ductal carcinoma in situ. Their respective morphologic character, biologic significance and natural history are discussed. Borderline lesions, such as atypical lobular hyperplasia and atypical ductal hyperplasia, are defined and their importance as risk factors for subsequent carcinoma is emphasized. Analysis of the concept of fibrocystic disease recognizes a lack of both pathologic specificity and predictive value as a risk factor for malignant disease. Rather than referring to fibrocystic disease, this complex should be stratified in specific histologic types to understand better its biologic significance. PMID- 2986808 TI - Minimal breast cancer: advance or anachronism? AB - The author argues that minimal breast cancer does not represent a distinct disease entity but comprises three discrete components, each with unique characteristics. The natural history of lobular carcinoma in situ and intraductal cancer, two of the components of minimal breast cancer, is described. The dangers of relying on a data base established on the strength of retrospective anecdotal information is underscored. Lobular carcinoma in situ has a propensity for multicentricity and bilaterality. In 25% of patients with lobular carcinoma in situ, invasive breast cancer will develop subsequently, and the majority of these tumours will be of ductal origin. The average interval from the diagnosis of lobular carcinoma in situ to the development of subsequent invasive cancer is over 15 years and both breasts are at equal risk. Based on this information, the use of bilateral prophylactic mastectomy is unjustified. In contrast to lobular carcinoma in situ, 25% to 50% of patients with intraductal carcinoma will subsequently have infiltrating cancer, at an average of 10 years after the initial biopsy. Although the putative incidence of multicentricity is 50%, virtually all subsequent invasive cancers occur not only in the same breast but in the same quadrant as the initial lesion. In light of the momentum for breast preserving operations in invasive cancer, clinical trials should be implemented to assess the propriety of conservative management with and without radiotherapy in patients with intraductal carcinoma. PMID- 2986809 TI - Surgical management of the patient at high risk for breast cancer. AB - The most controversial aspect of breast disease centres around the management of patients who have either a strong family history of breast cancer or a biopsy diagnosis of lobular carcinoma in situ or ductal carcinoma in situ. The current alternatives for patients who have two or more relatives with breast cancer consist of close follow-up or prophylactic total mastectomies and reconstruction. Invasive breast cancer in patients with lobular carcinoma in situ may occur in either breast and may be as high as 30% at 20 to 30 years. In these women it is reasonable to do a wide excision of the lobular carcinoma; in those without a family history, close follow-up is adequate. Intraductal cancer treated by biopsy only is associated with a 40% risk of cancer in the ipsilateral breast. Therefore, the usual management is total mastectomy. However, the information to support this therapy over a segmental resection has limited scientific validity. Because the cosmetic appearance after total mastectomy and reconstruction is not as good as that of the normal breast, this procedure must be employed cautiously and only with the total support of the patient and her husband or close family. Subcutaneous mastectomy for prophylaxis leaves behind macroscopic glandular tissue and, therefore, is not considered by many to be optimal management. A total mastectomy, preserving the skin and resecting all macroscopic breast tissue and nipple, is the treatment of choice if the procedure is deemed appropriate. PMID- 2986810 TI - Possible origin of human AIDS. PMID- 2986811 TI - Lung cancer following therapy for Hodgkin's disease. PMID- 2986812 TI - Outbreak of diarrhea in a day nursery in Ontario. PMID- 2986814 TI - Cushing disease presenting as atypical psychosis followed by sudden death. AB - The authors report a case of Cushing's Disease that first presented as a diagnostically confusing Atypical Psychosis. Sudden death occurred secondary to unexpected gastrointestinal bleed. Pathological findings confirmed the diagnosis. Neurochemical mechanisms for the behavioral aberrations are explored. PMID- 2986815 TI - Establishment and characterization of neoplastic cells from a malignant fibrous histiocytoma. A possible stem cell line. AB - Using the semisolid agar method, neoplastic three clones were isolated from a malignant fibrous histiocytoma. All clones represented morphologically only one type of cells having fine structure similar to that of histiocyte with multiple filopodia. The clones carrying Fc- and C3-surface receptors showed marked immunophagocytosis. They were positive for acid phosphatase, nonspecific esterase, and alpha-1-antitrypsin. These clones were able to display the potential for production of collagenous matrix. Moreover, inoculations of the each clone into nude mice resulted in productions of malignant fibrous histiocytoma with pleomorphic pattern. These tumors were composed morphologically of various types of cells such as immature, histiocyte-like, fibroblast-like, and multinucleated giant cells. These morphologic alterations of histiocytes occurred in all of the three clones in vitro. These findings suggest that the cloned cells established from the malignant fibrous histiocytoma are neoplastic histiocytes with capability to form various types of cells, a possible stem cell. PMID- 2986816 TI - Intraductal signet ring carcinoma. A hitherto undescribed form of intraductal carcinoma of the breast. AB - Attention is directed to a unique but apparently heretofore undescribed and overlooked form of intraductal carcinoma of female breast characterized by varying numbers of intraductal neoplastic cells with eccentric nuclei and either ground-glass or optically clear, mucin-containing cytoplasm. Eight examples have been observed in breast tissue submitted consultatively for the distinction of hyperplasia and intraductal carcinoma. This lesion, designated as intraductal signet ring carcinoma, may be found in some of the ducts of otherwise benign fibrocystic disease and radial scars, as well as breasts with other forms of intraductal carcinoma or invasive carcinoma. Four hundred consecutive examples of the latter examined prospectively revealed this intraductal lesion in 12 or 3%. Only 3 of the 12 examples were associated with either pure or combination forms of lobular invasive carcinoma. PMID- 2986813 TI - Mechanisms of virus-induced immune suppression. AB - The recent demonstration that the acquired immune deficiency syndrome (AIDS) is caused by a retrovirus that affects humans has given rise to widespread concern about the immunosuppressive properties of viruses in general. A wide variety of viruses have been shown to be able to compromise immune function. Sometimes immunosuppression results from the pathologic processes that viruses are able to induce. In other instances virus-induced immune derangements may themselves be responsible for the onset of pathologic change. In some cases a single infectious viral agent may be able to modulate several immunologic mechanisms simultaneously. This review discusses some of the various complex mechanisms through which viral infections can alter the function of the immune system. PMID- 2986817 TI - Induction of other differentiation stages in neoplastic epithelial duct and myoepithelial cells from the human salivary gland grown in athymic nude mice. AB - The interaction between two cell lines derived from the human salivary gland (HSG), neoplastic epithelial duct HSG cells and myoepithelial human pleomorphic adenoma (HPA) cells, was studied morphologically and immunohistochemically in nude mice tumors produced by inoculation of HSG and HPA cells. Transplantation of HSG cells into nude mice resulted in the production of adenocarcinoma which contained carcinoembryonic antigen (CEA). The nude mice tumors induced by HPA cells were interpreted as myoepithelioma in which the presence of S-100 protein and myosin were identified. On the other hand, the occurrence of squamous cell nests was frequently noted in the nude mice tumors produced by inoculation of a mixture of HSG and HPA cells. The tumor cells present in the squamous cell nests had abundant tonofilaments in the cytoplasm and were attached with tight junction and distinct desmosomes. In addition, the presence of keratin in the tumor cells composing squamous cell nests was demonstrated. When the mixture of HSG and HPA cells treated with polyethylene glycol (PEG) was transplanted into the nude mice, the tumors produced consisted almost entirely of areas showing the histologic features of anaplastic carcinoma, and did not contain all of the specific cell markers observed in the HSG or HPA tumors. The nude mice tumors induced by PEG treated HSG or HPA cells were interpreted as adenocarcinoma and myoepithelioma, respectively, and giant cells were occasionally observed in the tumor sections. These findings indicate that neoplastic cells showing differentiation stages other than those of the original two cells can be induced in nude mice by utilizing HSG and HPA cells. PMID- 2986819 TI - Biochemical and histochemical analysis of steroid hormone binding sites in human primary breast cancer. AB - Mammary carcinoma tissue from 514 primary breast cancer patients were all biochemically and histochemically analyzed for both estrogen receptors and progesterone receptors. The dextran-coated charcoal (DCC) method measured the ER and PR as defined by Scatchard analysis, ligand competition experiments and target organ specificity. The ligands, estradiol-6-carboxymethyloxime-BSA fluoresceine isothiocyanate and hydroxyprogesteronehemisuccinate-BSA tetramethylrhodamine isothiocyanate, used for histochemistry, did not bind to either ER or PR and were mainly bound to the membrane fraction of isolated breast cancer cells. Fluorescence was not specifically inhibited by estrogens or progestogens. In addition, "estrogenic" always coincided with "progestogenic" fluorescence. The binding of the fluoresceine compounds to tissue slides depended on the large steroid hormone substitution on the bovine serum albumin molecule. Clinical parameters, known to be related to ER and PR did not correlate with the histochemical results. The observations indicated the impossibility of specific steroid receptor detection by the histochemical method. Therefore, up to the present, evaluation of hormone dependency and prognosis in human breast cancer cannot be based on this approach. PMID- 2986818 TI - Karyometric analysis of liver cell dysplasia and hepatocellular carcinoma. Evidence against precancerous nature of liver cell dysplasia. AB - Karyometric analysis was performed with respect to anisonucleosis, nuclear deformity and DNA content in cases of liver cell dysplasia (LCD) and hepatocellular carcinoma (HCC). Presence or absence of iron deposition in foci of LCD and HCC was also evaluated in siderotic livers. All LCDs showed marked anisonucleosis and marked increases in DNA content but slight increases in nuclear deformity. A tendency was noted in which the nuclear deformity was increased as the nuclei became larger. In contrast, HCCs showed wide ranges of anisonucleosis, nuclear deformity and DNA content. Hepatocellular carcinomas having marked anisonucleosis similar to that of LCD showed markedly increased nuclear deformity. However, unlike LCD, this increase was independent of their nuclear size. In the siderotic livers iron deposition was noted in the foci of LCD but not in the foci of HCC. These findings do not support the notion of LCD being precancerous. PMID- 2986820 TI - CEA tissue staining in colorectal cancer patients. A way to improve the usefulness of serial serum CEA evaluation. AB - The evaluation of serial plasma carcinoembryonic antigen (CEA) levels is one of the most important parameter used to establish the prognosis of surgically cured colorectal cancer patients. Carcinoembryonic antigen is particularly useful in the identification of recurrences and metastasis. However, to improve the usefulness of this assay, it would be helpful to accurately determine, if possible, those patients whose cancers produce CEA. The evaluation of the presence of CEA in these cancer specimens by means of immunoperoxidase staining technique does seem to improve the sensitivity of the CEA test. Fifty-seven patients with colorectal cancer who underwent surgical treatment were studied. Tissue CEA evaluation was correlated with the plasma CEA levels, the pathologic stage and grade, and histologic type of the cancers. Results demonstrate that 66.6% of Dukes' B cancers, 78.9% of Dukes' C, and 77.7% of Dukes' D cancers stained positively for CEA by immunoperoxidase. Thirty of 57 patients with preoperative pathologic plasma CEA levels had positive tissue CEA, whereas 8/57 patients did not. Of patients with a well-differentiated cancer (G1), 81.4% had positive tissue CEA versus the 64% of G2 and 60% of G3 cancers. The authors conclude that the use of the immunoperoxidase stain to measure CEA in tissue, so that the CEA serum assay may be used in those patients known to produce CEA, results in a major increase in the sensitivity of the test. PMID- 2986821 TI - Atypical hyperplastic lesions of the female breast. A long-term follow-up study. AB - A total of 10,542 breast biopsy specimens obtained between 1950 and 1968 were studied. Examples of atypical "ductal" (ADH) and atypical lobular hyperplasia (ALH), defined as having only some features of carcinoma in situ (CIS), were diagnosed in 3.6% of these specimens. In the same series, CIS was diagnosed in 1.7% of biopsy specimens excluding those with invasive cancer. The subsequent risk of invasive breast carcinoma after ALH or ADH was 4-5 times that of the general population. Follow-up was 90% successful and extended 17 years after biopsy. History of breast cancer in a mother, sister, or daughter doubled the risk of subsequent invasive carcinoma development (to 8 times for ALH and 10 times for ADH). The authors conclude that among the epithelial hyperplastic lesions of the human breast, a minority may be recognized by their resemblance to CIS which have a clinically significant elevation of subsequent breast cancer risk. This risk is one-half that of CIS. PMID- 2986822 TI - Carcinoma of the breast in three siblings. AB - Reported are three siblings, a sister and 2 brothers, who had breast cancer. The paternal grandmother of the proband was known to have had breast cancer. It is our belief that this report supports further the genetic etiology of certain breast cancers. The study examines the literature with regard to genetically transmitted female breast cancer and also implies a genetic etiology of male breast cancer. Of special interest is the fact that the female sibling had bilateral breast cancer during her premenopausal years and that one of the two brothers also developed his lesion at an early age, 41 years. PMID- 2986823 TI - Levamisole in the treatment of stage II breast cancer. Five-year follow-up of a randomized double-blind study. AB - The effect of levamisole combined with postoperative radiotherapy in Stage II breast cancer was investigated in a double-blind randomized study comprising 72 patients. All patients were followed for at least 5 years. Disease-free survival was slightly prolonged in the levamisole group as a whole. Among postmenopausal patients, levamisole significantly increased both disease-free and total survival (P = 0.003) and P = 0.008, respectively). The levamisole group also showed fewer distant metastases as the first sign of recurrence. Levamisole treatment was associated with a risk of granulocytopenia and agranulocytosis (10%), but, as in the authors' previous studies, this seemed to be totally reversible and did not worsen the prognosis. PMID- 2986824 TI - Chemotherapy-induced remission in a patient with small cell carcinoma of the lung and hairy cell leukemia. AB - Small cell carcinoma of the lung developed in a patient with previously diagnosed hairy cell leukemia. Treatment with aggressive chemotherapy resulted in a complete response in both diseases lasting 13 months. Recurrence of the leukemia did not occur. This case demonstrates that hairy cell leukemia may be responsive to combination regimens. PMID- 2986825 TI - Malignant fibrous histiocytoma of bone. A review of 13 cases and an ultrastructural study. AB - Observations on 13 patients with primary malignant fibrous histiocytoma of bone were reported. Included were nine male and four female patients, ranging in age from 6 to 81 years (mean, 44.8 years). Pain was the most common complaint. The interval from the first symptom to the initial treatment varied from 2 months to 20 years. Seven tumors arose in the knee region. Roentgenologically, most of the lesions presented with an osteolytic and destructive appearance. Histologically, highly variable morphologic features existed. The storiform-pleomorphic pattern was found in every tumor, although it was not necessarily pathognomonic for malignant fibrous histiocytoma of bone. Surgery, radical or incomplete, was the primary treatment for all but one patient. Lymph node metastasis was present in three. Five patients died of the disease from 3 to 79 months after the diagnosis (mean, 28 months), all exhibiting metastasis to the lung. Gaucher's body-like structure demonstrated in the electron microscopic study shows the histiocytic quality of malignant fibrous histiocytoma of bone. PMID- 2986826 TI - Transcatheter arterial embolization for hepatocellular carcinoma. Benefits and limitations for unresectable cases with liver cirrhosis evaluated by comparison with other conservative treatments. AB - Transcatheter arterial embolization (TAE) for unresectable hepatocellular carcinoma (HCC) was performed in cirrhotic patients with nonoccluded main portal trunk and total bilirubin levels less than 5 mg/dl. In 48 patients with tumor response to TAE, its benefit was evaluated by comparing their survival with that in 28 patients after conservative treatments, matched by clinical data. In TAE treated patients, tumor findings on computerized tomography and angiogram were significantly different between those dying within 3 months and those surviving over 18 months. In cases of tumors occluding the first- or second-order portal branches or occupying more than 20% of the liver, or relatively hypovascular tumors with unclear boundaries, survival was not different between TAE- and non TAE-treated patients, whereas it was significantly improved by TAE in cases without these findings. These results suggest that TAE is beneficial in cases of unresectable HCC complicating liver cirrhosis, but its benefit is limited under certain conditions. PMID- 2986828 TI - Long-term survival of patients with hepatocellular carcinoma combined with liver cirrhosis. Report of two patients. AB - Two patients with hepatocellular carcinoma combined with liver cirrhosis lived more than 5 years after hepatic resection. One patient with liver cirrhosis combined with schistosomiasis japonica survived 5 years and 2.5 months after left lobectomy and died of liver failure. Two recurrent tumors, each 1 cm in diameter, were present in the remaining lobe but were not responsible for death. Another patient is well 9 years and 2 months after extended right lobectomy. The tumor was bulky, and the resected specimen was 2800 g in weight. These facts show that patients with hepatocellular carcinoma combined with liver cirrhosis can survive for a long period even if a large tumor is involved. PMID- 2986827 TI - Hepatolithiasis associated with cholangiocarcinoma. Possible etiologic significance. AB - Three cases of primary bile duct carcinomas (cholangiocarcinomas) were found among 61 cases of hepatolithiasis. Cholangiocarcinoma arose from the extrahepatic bile duct in one and from the dilated intrahepatic bile duct in two patients. Hyperplasia of the columnar cells was often present. These hyperplastic epithelial cells often show papillomatous or adenomatous pattern, which are frequently associated with the presence of stones and the contaminated bile, and may show malignant changes leading to the development of cholangiocarcinoma. PMID- 2986829 TI - Lung cancer in women. Lahey Clinic experience, 1957-1980. AB - The incidence of lung cancer is increasing dramatically worldwide. Cancer of the lung, the number one cause of death from cancer in men in the United States, will soon surpass breast cancer to become the most frequent cause of cancer death in women. To detail the changing pattern of lung cancer, we reviewed the clinical features of all 1752 patients whose lung cancer was diagnosed at the Lahey Clinic from 1957 through 1980. Women comprised a constant proportion of the total number of clinic patients during this 24-year period. Lung cancer in women increased markedly from 13% of all patients (1 to 6.8 ratio of women to men) in 1957 through 1960 to 35% (1 to 1.8 ratio) in 1977 through 1980. The authors reviewed pathologic specimens of 394 women with lung cancer. No significant change occurred in cell type distribution over the years: adenocarcinoma, 38%; epidermoid carcinoma, 20%; large cell carcinoma, 15%; small cell undifferentiated tumor, 13%; and other cell types, 14%. The incidence of all lung cancer cell types (Kreyberg Group 1 and Group 2) increased in women who smoked. Our study suggests that smoking is a major causal factor in the rising occurrence of all lung cancer cell types in women as contrasted to the preponderance of Kreyberg Group 1 tumors in men who smoke. PMID- 2986830 TI - Cytoskeletal events during calcium- or EGF-induced initiation of DNA synthesis in cultured cells. Role of protein phosphorylation and clues in the transformation process. AB - The possible relationship between cytoskeletal events and growth regulation in response to stimulation by calcium and by growth factors such as EGF can be summarized as follows: An elaborate interaction exists between calcium and serum growth factors, such as EGF, in the initiation of DNA synthesis in quiescent cells. This implies that many processes between the external signals delivered at the cell surface and the sequential intracellular events that lead to chromosomal replication, and ultimately to cell division, must be coordinated in a reproducible manner. It is now apparent that because of its possible role as a dynamic integrator of the cytoplasm, the cytoskeleton could represent the coordinator of the events that lead to replication. Calcium (with its intracellular acceptor, calmodulin) and cAMP (which can act by opposing mechanisms) are extensively involved in the control of the integrity of the cytoskeleton. Distinct protein kinases are activated by calcium/calmodulin, EGF, and cAMP as aspects of the prereplicative response, and many of the substrates for phosphorylation are cytoskeletal proteins. The emerging picture seems to include a direct involvement of these protein kinases in the cascade of regulatory events that leads to the initiation of DNA synthesis. Thus, the cytoskeleton has a direct role in the transmission of proliferative signals from external receptor sites to the nucleus. A means by which neoplastic cells can bypass the normal regulatory pathways is proposed in the light of recent data showing that the product of oncogenes are protein kinases or proteins that intimately interact with cellular protein kinases. PMID- 2986832 TI - Biotransformation and protein binding of N-(4-hydroxyphenyl)retinamide in murine mammary epithelial cells. AB - The biotransformation of N-(4-hydroxyphenyl)retinamide (HPR), and interactions of parent compound and/or metabolites with the cellular retinoid binding proteins (CRBPs) and cellular retinoic acid binding proteins (CRABPs) were examined in murine mammary tumor virus (MuMTV)-induced murine mammary tumor cells (GR-3A) grown in monolayer cell culture. Soluble fractions (cytosols) obtained from the extracts of GR-3A cells after high speed centrifugation were found to contain proteins of approx. 15,000 daltons which bound retinol and retinoic acid, but did not bind HPR or HPR metabolites. Moreover, HPLC analysis of GR-3A cell extracts demonstrated that [3H]retinol and [3H]retinoic acid were not detected in cells that had been exposed to [3H]HPR for 48 h. These findings, that under in vitro conditions there is no appreciable enzymatic hydrolysis of HPR to retinoic acid or conversion to retinol, suggests that the metabolism and cytological effects of HPR may be distinct from those of retinol or retinoic acid within murine mammary epithelial cells. PMID- 2986833 TI - Etoposide and cisplatin salvage chemotherapy for small cell lung cancer. AB - Thirty patients with previously treated small cell lung cancer received salvage combination chemotherapy with etoposide and cisplatin. Two complete and six partial responses were observed, for a major response rate of 27%. Responses occurred promptly and sustained palliation was achieved among responders. Myelosuppression was the major dose-limiting toxic effect. A schedule of etoposide (115 mg/m2 iv on Days 1-3) and cisplatin (25 mg/m2 iv on Days 1-3 every 28 days) is recommended for further clinical trials. PMID- 2986831 TI - Molecular and cytologic analysis of DNA amplification in retinoblastoma. AB - Amplification of two distinct genomic DNA segments is observed in homogeneously staining regions in two sets of retinoblastoma cell lines derived from two different patients. One DNA segment was known to have sequence homology to the c myc oncogene, and both DNA segments had previously been shown to be amplified in neuroblastoma cells. The absolute degree of amplification differed in all cytogenetically distinct retinoblastoma cell lines tested. Also, the relative amplification of these two DNA segments was unequal within a given cell line. Minimal amplification of both DNA segments was also detected in DNA directly isolated from one primary retinoblastoma. Based on these and previous results, it is concluded that assembly of amplifiable, relocatable units in many human retinoblastoma and neuroblastoma cells may involve a complex process of differential recruitment of separate DNA segments that are located on human chromosome #2. PMID- 2986834 TI - Phase II trial of 1,2-diaminocyclohexane-(4-carboxyphthalato) platinum(II) in patients with refractory germ cell tumors. PMID- 2986835 TI - Phase II trial of mitoguazone in patients with refractory germ cell tumors. PMID- 2986836 TI - Structure and behaviour of Stannius corpuscles in relation to vitamin D3 induced hypercalcemia in male Clarias batrachus. PMID- 2986837 TI - Immunoelectron microscopy of adult T-cell leukemia-associated antigen by the peroxidase-labeled antibody method. PMID- 2986838 TI - Immunoelectron microscopic detection of surface antigen of simian virus 40 transformed cells. PMID- 2986839 TI - Fine structural and immunohistochemical studies of goat adenohypophysial cells. AB - The fine structure of each type of anterior pituitary cell in the male goat was studied through the application of a superimposition technique in which adjacent thick sections were used to identify individual cells beforehand by light microscopic immunohistochemistry. A cone of the pars intermedia protrudes into the pars anterior, being surrounded by the narrow pituitary cleft; the immunohistochemical appearances of the cells forming the cone resemble those of the pars anterior. Several follicles appear in the pars anterior. Ultrastructurally GH cells resemble prolactin cells. The secretory granules of both types are spherical; the diameter of the former is about 340 nm, whereas that of the latter is about 440 nm. ACTH cells are polygonal in shape with secretory granules, about 180 nm in diameter, scattered throughout the cytoplasm. TSH cells, which are spherical in shape, contain the smallest secretory granules, 150 nm in diameter. The highly electron-dense LH cells contain numerous secretory granules about 210 nm in diameter. Their nuclei are irregular with incisures. Thus, the anterior pituitary cells of the goat are ultrastructurally characteristics and species-specific. PMID- 2986840 TI - A point mutation in the conserved hexanucleotide at a yeast 5' splice junction uncouples recognition, cleavage, and ligation. AB - We have constructed an actin-HIS4 gene fusion, such that expression of HIS4 requires proper splicing of the actin intron. Using this chimeric gene in an in vivo screen for splicing mutations, we have isolated a G to A transition in the fifth position of the yeast 5' consensus sequence/GTAPyGT. This mutation still allows the junction to be recognized by the splicing machinery, albeit inefficiently. Surprisingly, the fidelity of the 5' endonucleolytic cleavage is also reduced. This results in an incorrect cleavage 6 nucleotides 5' of the 5' junction, at the dinucleotide/AT. Cleavage at this abnormal site does not lead to the production of mature mRNA, although this species appears to be in a lariat structure. The behavior of this mutant argues that recognition of the 5' junction and subsequent cleavage are separable events and, furthermore, that requirements for 3' endonucleolytic cleavage may be more complex than previously imagined. PMID- 2986841 TI - In vivo mapping of DNA topoisomerase II-specific cleavage sites on SV40 chromatin. AB - The antitumor drug, m-AMSA (4'-(9-acridinylamino)-methanesulfon-m-anisidide), is known to interfere with the breakage-reunion reaction of mammalian DNA topoisomerase II by blocking the enzyme-DNA complex in its putative cleavable state. Treatment of SV40 virus infected monkey cells with m-AMSA resulted in both single- and double-stranded breaks on SV40 viral chromatin. These strand breaks are unusual because they are covalently associated with protein. Immunoprecipitation results suggest that the covalently linked protein is DNA topoisomerase II. These results are consistent with the proposal that the drug action in vivo involves the stabilization of a cleavable complex between topoisomerase II and DNA in chromatin. Mapping of these double-stranded breaks on SV40 viral DNA revealed multiple topoisomerase II cleavage sites. A major topoisomerase II cleavage site was preferentially induced during late infection and was mapped in the DNAase I hypersensitive region of SV40 chromatin. PMID- 2986842 TI - The yeast nuclear gene NAM2 is essential for mitochondrial DNA integrity and can cure a mitochondrial RNA-maturase deficiency. AB - Dominant mutations in the yeast nuclear gene NAM2 cure the RNA splicing deficiency resulting from the inactivation of the bI4 maturase encoded by the fourth intron of the mitochondrial cytochrome b gene. This maturase is required to splice the fourth intron of this gene and to splice the fourth intron of the mitochondrial gene oxi3 encoding cytochrome oxidase subunit I. We have cloned the nuclear gene NAM2, which codes for two overlapping RNAs, 3.2 kb and 3.0 kb long, which are transcribed in the same direction but differ at their 5' ends. NAM2 compensating mutations probably result from point mutations in the structural gene. Integration of the cloned gene occurs at its homologous locus on the right arm of chromosome XII. Inactivation of the NAM2 gene either by transplacement with a deleted copy of the gene, or by disruption, is not lethal to the cell, but leads to the destruction of the mitochondrial genome with the production of 100% cytoplasmic petites. PMID- 2986843 TI - Interaction of specific nuclear factors with the nuclease-hypersensitive region of the chicken adult beta-globin gene: nature of the binding domain. AB - We have studied the properties of a factor or factors that bind selectively to the 5' flanking region of the chicken adult beta-globin (beta A-globin) gene. We previously showed that these components, when bound with histones on plasmids containing the region, confer on the complex a pattern of hypersensitivity to nuclease digestion similar to that in the nucleus. We have now measured the abundance of the factor(s) in partially purified preparations, and the affinity constants for binding to specific and nonspecific DNA sequences. Footprinting studies of the specific complex with DNAase I and II reveal two discrete protected regions within the hypersensitive domain. When these regions are physically separated, they interact with the factor(s) independently, suggesting that each region binds one or more distinct components. The footprint patterns of our complexes in vitro agree with the patterns observed in intact chicken erythrocyte nuclei. These complexes thus are on the transcriptionally active beta A-globin gene in vivo. PMID- 2986844 TI - Role of the nucleocapsid protein in regulating vesicular stomatitis virus RNA synthesis. AB - We describe experiments with two monoclonal antibodies to the vesicular stomatitis virus (VSV) nucleocapsid protein N with strikingly different characteristics. Antibody 1 binds to nucleocapsids and probably the pool of free (unbound) N protein; it inhibits transcription in vitro, and when microinjected into cells, protects the cells against VSV. Antibody 2 binds poorly to nucleocapsids, does not inhibit transcription, but when microinjected into cells, binds selectively to the free N and delays the appearance of progeny virus. We have confirmed these results by analyzing the effect of these antibodies on in vitro genomic RNA synthesis. The results of both the in vivo and in vitro experiments show that the replication of the VSV genome is controlled by the availability of the nucleocapsid protein, even when the polymerase has access to the host factors and multiple phosphorylated forms of the NS protein thought to be involved in genomic RNA synthesis. PMID- 2986845 TI - Sequence determination of the Sendai virus HN gene and its comparison to the influenza virus glycoproteins. AB - The nucleotide sequence of the Sendai virus (SV) HN (hemagglutinin-neuraminidase) gene was determined. The deduced primary structure of the protein showed only one hydrophobic domain likely to represent the transmembrane region, but at its N terminus. Since the SV F protein is anchored in the membrane at its C terminus, the two SV glycoproteins are thus membrane-anchored in opposite orientations, similar to the two influenza virus (FLU) glycoproteins. Amino acid sequence comparisons of the SV HN and the FLU HA and NA proteins revealed homologies between 100 amino acids of the hemagglutinin region of the FLU HA protein and the C terminus of the SV HN, and between 200 amino acids of the neuraminidase region of the FLU NA and the central region of SV HN. Alignment of the neuraminidase, hemagglutinin, and fusion regions shared by these glycoproteins suggest the structure of a possible ancestral gene. PMID- 2986846 TI - Rearrangements of viral and cellular DNA are often associated with expression of Rous sarcoma virus in rat cells. AB - Rous sarcoma virus (RSV) proviruses integrated within the DNA of transformed rat cells frequently display duplications of variable segments of proviral DNA upstream of an intact provirus. The rearrangement in the A11 clone of transformed rat cells consists of a partial duplication of both viral and cellular DNA segments whose origin is a region of approximately 4 kb encompassing the 3' virus cell junction. Transposition of this DNA appears to have occurred at or after virus integration by a mechanism involving at least two recombination events. In every case examined, including A11, the transcriptional organization of the original provirus has been conserved and viral RNA expression appears to occur normally. The frequency of such rearranged proviruses in the DNA of transformed rat cells suggests that upstream rearrangements may influence provirus expression. PMID- 2986847 TI - Transcription control by oncogenes. PMID- 2986848 TI - Coexpression of the sis and myc proto-oncogenes in developing human placenta suggests autocrine control of trophoblast growth. AB - First trimester human placentas actively express the sis proto-oncogene, the structural gene for the B chain of platelet-derived growth factor (PDGF). Using the in situ hybridization technique, the 4.2 kb c-sis transcript has been localized to the cytotrophoblastic component, especially the highly proliferative and invasive cytotrophoblastic shell, paralleling the distribution of c-myc transcripts in early placenta. Explants of first trimester placenta release significant levels of PDGF-like activity into the medium under apparent developmental control. Moreover, cultured trophoblasts display abundant high affinity PDGF receptors and respond to exogenous authentic PDGF by an activation of the c-myc gene and DNA synthesis. The developing human placenta may therefore represent a case of autocrine growth regulation in a normal tissue, in which cells bearing receptors for a growth factor can also synthesize and respond to that factor. PMID- 2986850 TI - An insertion within a variably spliced Drosophila tropomyosin gene blocks accumulation of only one encoded isoform. AB - We have characterized an aberrant allele of a variably spliced Drosophila tropomyosin gene. The allele was recovered from the flightless Ifm(3)3 strain, which has been shown to have structurally and functionally abnormal indirect flight muscles. The transcribed portion of the mutant gene is interrupted by an 8,8 kb insertion of middle repetitive DNA having a structure typical of copia like Drosophila mobile elements. The insertion is positioned so as to interrupt an exon sequence in one splicing mode and, simultaneously, an intron in the alternate mode. As a consequence of the insertion the allele fails to direct synthesis of the flight muscle-specific tropomyosin isoform, but remains capable of specifying a second isoform, which accumulates in nonfibrillar Drosophila muscles. PMID- 2986849 TI - O-linked oligosaccharides of mouse egg ZP3 account for its sperm receptor activity. AB - Previously, we reported that ZP3, one of three different glycoproteins present in the mouse egg's zona pellucida, serves as a sperm receptor. Furthermore, small glycopeptides derived from egg ZP3 retain full sperm receptor activity, suggesting a role for carbohydrate, rather than polypeptide chain in receptor function. Here, we report that removal of O-linked oligosaccharides from ZP3 destroys its sperm receptor activity, whereas removal of N-linked oligosaccharides has no effect. A specific size class of O-linked oligosaccharides, recovered following mild alkaline hydrolysis and reduction of ZP3, is shown to possess sperm receptor activity and to bind to sperm. The results presented strongly suggest that mouse sperm bind to eggs via O-linked oligosaccharides present on ZP3. PMID- 2986851 TI - Intricate combinatorial patterns of exon splicing generate multiple regulated troponin T isoforms from a single gene. AB - Mechanisms of alternative RNA splicing, important in the generation of protein diversity, are common but incompletely understood. Among the contractile proteins, troponin T exists in several isoforms, shown to be derived in part from a novel pattern of differential RNA splicing in the 3' region of the rat skeletal fast troponin T gene. In fact, this gene has a previously unsuspected capacity to encode multiple isoforms. The isolation of four distinct but related cDNAs from this gene, which share discontinuous subsegments of sequence identity in their 5' regions, and the determination of the genomic sequence, demonstrate that small exons with characteristic split codon structure are differentially spliced in intricate combinatorial patterns to generate a minimum of 10, and potentially 64, distinct troponin T mRNAs, encoding different isoforms, in a developmentally regulated and tissue-specific manner. At least two of these mRNAs are spliced from structurally identical primary transcripts, necessitating control by trans acting factors. PMID- 2986852 TI - Increase of vulnerability to lymphotoxin in cells infected by vesicular stomatitis virus and its further augmentation by interferon. AB - The cytotoxic effect of lymphotoxin (LT) and its modulation by interferon (IFN) was quantitatively assessed in uninfected and vesicular stomatitis virus (VSV) infected cultured cells. Preparations of human LT, which were depleted of IFN, had a significant cytotoxic effect on VSV-infected HeLa, SV-80, WISH, and Vero cells. IFN, most notably IFN-gamma, further potentiated destruction of the infected cells by these LT preparations, when applied on the cells at sub antiviral IFN concentrations. In contrast, no cytotoxic effect could be observed in any of the examined cells, when applying LT, IFN, or their combination, in the absence of viral infection. Infected cells in which VSV replication was suppressed by treatment with antiviral concentrations of IFN also resisted destruction by LT. These findings indicate that LT cytotoxicity can be selectively directed against virus-infected cells and that IFN can augment this cell-killing mechanism when failing to exert an antiviral effect. PMID- 2986854 TI - Products of the lipoxygenase pathway in human natural killer cell cytotoxicity. AB - As earlier data suggested the importance of lipoxygenase activation for expression of human NK cell cytotoxicity, four different lipoxygenase inhibitors were tested for suppression of natural killer (NK) cell lysis. All inhibitors were found active at nontoxic concentrations with 50% inhibition at approximately 15 microM for nordihydroguaiaretic acid (NDGA). NK cell lysis could be reconstituted to NDGA-suppressed cells with leukotriene B4 (LTB4), the all-trans isomers 6-trans-LTB4 and 12-epi-6-trans-LTB4, and 20-COOH-LTB4. LTB4 reconstitution was best in the concentration range 1-100 pM and near control levels at both higher and lower concentrations. Herpesvirus Ateles-transformed killer T cells could also be inhibited by NDGA. These data indicate that lipoxygenase activity is required for human NK cell lysis and that several different LTB4-related products can restore NK activity in inhibited cells; they also suggest that the lipoxygenase pathway is present in the killer cell population. PMID- 2986853 TI - Regulation of macrophage collagenase, prostaglandin, and fibroblast-activating factor production by anti-inflammatory agents: different regulatory mechanisms for tissue injury and repair. AB - Activation of macrophages results in the production of tissue destructive mediators and enzymes including prostaglandins (PGE) and collagenase. In addition, activated macrophages also generate mediators which enhance connective tissue formation through their effects on fibroblast growth. To determine whether the pro-inflammatory mediators and the mediator(s) involved in tissue repair are under the same regulatory control, guinea pig macrophage cultures were treated with various pharmacologic agents and their supernatants monitored for biologic activity. The nonsteroidal anti-inflammatory agent, indomethacin, and the glucocorticoid, dexamethasone, at pharmacologic concentrations inhibited not only prostaglandin synthesis (greater than 90%) but also the production of collagenase (greater than 90%). Colchicine, a microtubule disruptive agent, but not the inactive form, lumicolchicine, markedly diminished the production of collagenase independently of prostaglandin synthesis. In contrast to the inhibitory effects of these anti-inflammatory agents on PGE and collagenase production, indomethacin did not inhibit the production of macrophage-derived fibroblast-activating factor (FAF). Furthermore, dexamethasone at pharmacologic doses did not inhibit FAF production. Colchicine not only did not inhibit FAF, but frequently enhanced the appearance of FAF In the macrophage cultures. Thus, it appears that regulation of the production of PGE and collagenase is different than the regulation of FAF synthesis and therefore the production of these mediators can be differentially modulated. Such a dissociation may provide a basis for mononuclear cell-mediated fibroblast growth and tissue repair to occur independently of the release of PGE2 and collagenase and even following anti-inflammatory drug therapy. PMID- 2986855 TI - Control of growth and expression of differentiated functions of mature hepatocytes in primary culture. AB - Since methods to disperse and culture hepatocytes were developed 15 years ago, numerous investigations have shown that primary cultures of mature hepatocytes retain most liver functions and respond as well to various hormones as those in vivo. Thus they are the most suitable system in vitro for studies on the liver. Moreover, recently it was found that differentiated hepatocytes in culture can grow under certain conditions and that this growth is regulated not only by several hormones, such as insulin, epidermal growth factor and serum growth factor, but also by a cell membrane factor and proteins in the environmental matrix through cell contact. This article describes the biochemical characterization of regulatory factors for hepatocyte growth and functions and their reciprocal expression. The mechanisms of liver regeneration, differentiation and carcinogenesis and the importance of the tissue architecture for these events are discussed mainly on the basis of our findings. PMID- 2986856 TI - [Extra-gonadal germinative tumors of the retroperitoneum. Case report]. PMID- 2986857 TI - Inhibition by reactive aldehydes of superoxide anion radical production in stimulated human neutrophils. AB - alpha,beta-Unsaturated aldehydes were investigated in vitro for their ability to inhibit superoxide anion radical (O2-.) production in stimulated human polymorphonuclear leukocytes (PMN). The aldehydes investigated were (i) trans-4 hydroxynonenal and malonaldehyde (MDA), two toxic lipid peroxidation products; (ii) acrolein and crotonaldehyde, two air pollutants derived from fossil fuel combustion; (iii) trans,trans-muconaldehyde, a putative hematotoxic benzene metabolite. Preincubation of PMN with reactive aldehydes followed by stimulation with the oxygen burst initiator phorbol myristate acetate (PMA) resulted in a dose-dependent inhibition of O2-. production. The concentration at which 50% inhibition (IC50) was observed was 21 microM for acrolein, 23 microM for trans,trans-muconaldehyde, 27 microM for trans-4-hydroxynonenal and 330 microM for crotonaldehyde. A similar inhibitory effect by these aldehydes was observed in digitonin- and concanavalin A-stimulated PMN. MDA inhibited O2-. production in PMA-stimulated PMN by 100% at 10(-2) M but gave no inhibition at 10(-3) M. The standard aldehyde propionaldehyde did not inhibit O2-. production at 10(-3)-10( 6) M. Preincubation of PMN with acrolein in the presence of cysteine completely protected against the inhibitory effect of this reactive aldehyde. The results indicate that the ability of toxic aldehydes to inhibit O2-. production in stimulated PMN correlates directly with their alkylation potential which is a function of the electrophilicity of the beta carbon. PMID- 2986858 TI - Effects of 4-hydroxynonenal on adenylate cyclase and 5'-nucleotidase activities in rat liver plasma membranes. AB - Adenylate cyclase and 5'-nucleotidase activities in rat liver plasma membranes were assayed in vitro in the presence of 4-hydroxy-2,3-trans-nonenal (HNE), a major end-product of microsomal lipid peroxidation. Both basal and glucagon stimulated adenylate cyclase were inhibited in a dose-dependent manner, even at micromolar HNE concentrations, whereas fluoride-stimulated activity increased. A biphasic, dose- and time-dependent effect was noted when the basal activity was monitored at increasing doses. 5'-Nucleotidase activity was also decreased by HNE, but only at millimolar concentrations. These findings are related to the view that aldehydes, especially HNE, may act as diffusible cytotoxic compounds when lipid peroxidative derangement of membrane lipids is provoked by toxic conditions. PMID- 2986859 TI - Interaction of intercalating and non-intercalating agents with DNA: use of hydroxyapatite chromatography and S1 nuclease. AB - We have used hydroxyapatite (HA) chromatography and S1 nuclease hydrolysis to study the modification in the secondary structure of DNA caused by certain intercalating and non-intercalating ligands. The principal conclusions of HA experiments were as follows: (1) when native DNA, complexed with drugs believed to bind to DNA by intercalation (ethidium bromide, acridine orange, actinomycin D and acriflavin), is chromatographed on HA a lower affinity of DNA for HA is observed; also, the DNA elutes from HA columns as a drug-DNA complex; (ii) ligands that are known to interact with DNA by surface interactions do not show these effects; (iii) it may be possible to quantitate the binding of the intercalating drug to DNA and to determine its degree of binding by HA chromatography. Possibly, intercalation causes a change in the configuration of the sugarphosphate backbone of DNA, resulting in an altered steric orientation or 'burial' of phosphate groups with reduced availability for surface interactions with HA. S1 nuclease was used to determine the thermal melting profiles of DNA complexed with ethidium bromide and acridine orange. The melting profile in both cases was found to be biphasic with considerably reduced denaturation even at 95 degrees C. This is accounted for by the property of intercalating agents of stabilizing the secondary structure of DNA and the reported preference in binding to G-C base pairs. PMID- 2986860 TI - Peroxidase-mediated irreversible binding of arylamine carcinogens to DNA in intact polymorphonuclear leukocytes activated by a tumor promoter. AB - Addition of the tumor promoter phorbol myristate acetate to polymorphonuclear leukocytes results in the oxidation of the arylamine carcinogens; [14C]benzidine, N-[14C]methylaminoazobenzene and [14C]aminofluorene to reactive intermediate(s) that bind irreversibly to the leukocyte DNA. The binding was dependent on oxygen and was decreased by sulfhydryl inhibitors and phenolic antioxidants that inhibit the respiratory burst triggered by the phorbol myristate. Both the binding and the respiratory burst were increased by azide, presumably as a result of intracellular catalase inhibition. However higher concentrations of azide and cyanide prevented binding without affecting the respiratory burst indicating that myeloperoxidase is a catalyst for the binding. Granules isolated from the activated leukocytes and H2O2 catalyzed a cyanide sensitive benzidine binding to calf thymus DNA. Myeloperoxidase and H2O2 also catalysed extensive binding of these arylamines to calf thymus DNA. The leukocytes appear to be a useful model cell for studying one electron oxidation-catalyzed carcinogen activation. PMID- 2986862 TI - [Isolation and identification of the Alu family sequence in the human genome]. PMID- 2986863 TI - [Topological isomers of pBR322 DNA in vivo]. PMID- 2986861 TI - Opiate receptor interaction of a new analgesic drug 2-piperidinoethyl dibenzylglycolate. AB - The pharmacological properties of a new analgesic drug, 2-piperidinoethyl dibenzylglycolate (PDG), have been demonstrated by classical tests. The technique of iontophoresis was used in order to compare the effects of PDG with those of Tyr-D-Ser-Gly-Phe-Leu-Thr (DSTLE), syndyphalin, morphine and naloxone (NAL) on hypothalamic neurones. PDG as other four substances evoked only inhibitory responses. Some neurones, on which were tested three substances, were sensitive to one, two or three of these substances. The differential responses so obtained suggested that PDG does not act on mu- and delta-receptors but on an unidentified receptor for which morphine and NAL have a high affinity as agonist. Structural requirements for activity on different receptors were also proposed on the basis of crystallographic data and the above results. PMID- 2986864 TI - [Latency of HSV-1 in trigeminal ganglia of mice]. PMID- 2986865 TI - [Non-specific influx of cytotoxic T cells into influenza virus-infected lungs of mice]. PMID- 2986866 TI - [Molecular recloning of herpes simplex virus type I genome]. PMID- 2986867 TI - [Hepatoma initiating and promoting effects of gossypol]. PMID- 2986868 TI - [Study on heterogenicity of opiate receptors in the rat brain]. PMID- 2986869 TI - [Use of the 5406 bacterial fertilizer to increase production of ginseng]. PMID- 2986870 TI - [Clinical efficacy of aconitic injection in asthenia patients suffering from sick sinus syndrome and its mechanism]. PMID- 2986872 TI - [Experimental studies of the effects of huang wu san on microbial growth and viral infections]. PMID- 2986871 TI - [Zinc, copper and iron contents and their clinical significance in 40 kinds of Chinese traditional menses-regulating drugs]. PMID- 2986873 TI - [Possibilities for surgical excision of diffuse hepatoblastomas. Contribution and limitations of chemotherapy]. AB - Four children with major hepatoblastoma were treated with extensive surgical excision and chemotherapy. Two right lobectomies and two left extended hepatectomies were performed. The first patient only received postoperative chemotherapy. The others three had preoperative chemotherapy for unresectable hepatoblastoma. All these children exhibited an initial response with marked reduction in size of their primary tumor. The only child who died during operation had disease involving both lobes of the liver, inferior vena cava and lungs metastasis. The follow up of the three other children is 26 to 32 months. Two of them are free of all disease. The third had lung isolated metastasis removed and is actually free of disease with follow up of 32 months. Authors discuss chemotherapy interest and limits in major hepatoblastoma. PMID- 2986874 TI - Molecular characterization of adrenergic receptors. PMID- 2986875 TI - Acetylcholine release from rat atria can be regulated through an alpha 1 adrenergic receptor. AB - Isolated superfused rat atria release [3H]acetylcholine when depolarized with 57 mM potassium. The depolarization-induced overflow of [3H]acetylcholine is markedly inhibited by micromolar concentrations of epinephrine and norepinephrine. The alpha 1-selective adrenergic agonist methoxamine also inhibits tritium overflow, but the alpha 2-selective adrenergic agonist clonidine and the beta-adrenergic agonist isoproterenol do not. Prazosin, an selective alpha 1-adrenergic antagonist, blocks adrenergic inhibition of [3H]acetylcholine overflow with a Ki of approximately 0.4 nM. Yohimbine has approximately one hundredth the potency of prazosin for blocking adrenergic inhibition of [3H]acetylcholine overflow. [3H]Norepinephrine overflow from isolated rat atria is also inhibited by norepinephrine, but this effect is antagonized by yohimbine and not by prazosin. We suggest that the release of acetylcholine from cardiac parasympathetic neurons can be regulated through an alpha 1-adrenergic receptor, and that this mechanism may underly, at least in part, the relative lack of effects of prazosin on heart rate. PMID- 2986876 TI - Platelet activation and secretion associated with emotional stress. AB - Platelets are believed to play a role in the pathogenesis of atherosclerosis and of the vascular obstruction that causes the acute complications of coronary artery disease. Since specific behavioral patterns appear to be related to the development of coronary artery disease and since emotional stress may predispose an individual to acute cardiovascular ischemia, it was hypothesized that platelet activation by catecholamines might be involved in these events. To study emotional stress, plasma samples were obtained from 61 senior medical residents immediately before they were to speak in public. There were significant increases in the plasma concentrations of the platelet-secreted proteins platelet factor 4 and beta-thromboglobulin and epinephrine and norepinephrine immediately before speaking, which demonstrates that platelet activation and secretion occur in association with this type of emotional stress. Four trials were carried out to study the mechanism for this observed platelet secretion: (1) phenoxybenzamine, (2) propranolol, (3) 650 mg aspirin, and (4) 80 mg aspirin were given several hours before the public speaking engagement. Neither phenoxybenzamine nor propranolol in doses that blocked the hemodynamic effects of alpha 1- and beta 1 adrenergic stimulation modified platelet secretion. Aspirin also did not block platelet secretion, which suggests that platelets were not being stimulated through a cyclooxygenase-dependent pathway. This study provides direct evidence of platelet secretion in vivo in association with emotional stress, and underscores the potential importance of platelet activation and secretion in the acute events that occur in patients with vascular disease. PMID- 2986877 TI - Enhancement of the antiaggregatory activity of prostacyclin by propranolol in human platelets. AB - The beta-adrenergic antagonist propranolol has been found to inhibit platelet aggregation. We investigated the possibility that propranolol exerts this action by stimulating the synthesis or enhancing the antiaggregatory activity of prostaglandin (PG) I2. The media from cultures of human endothelial cells inhibited thrombin-induced platelet aggregation, an effect attributed to PGI2 production by the cells. When endothelial cells were incubated with dl- or d propranolol, the media had two to three times the inhibitory activity of control media. However, this increased activity was not due to increased synthesis of PGI2 because control and propranolol-treated cultures synthesized similar amounts of the PGI2 metabolite, 6-keto-PGF1 alpha. Instead, propranolol enhanced the antiaggregatory activity of PGI2. Propranolol (1 microM) and PGI2 (0.05 nM), when tested separately, inhibited aggregation by 19% and 13%, respectively, whereas the combination inhibited aggregation by 51%. PGI2 inhibited platelet aggregation and thromboxane (Tx) B2 production but stimulated cyclic AMP formation. The adenyl cyclase inhibitor 2',5'-dideoxyadenosine (DDA) had no effect of its own on these parameters, but blocked the actions of PGI2. Propranolol inhibited aggregation and TxB2 synthesis without changing cyclic AMP levels. Unlike PGI2, propranolol's effects were not altered by DDA. While the combination of propranolol and PGI2 inhibited aggregation to a greater extent than either agent alone, this enhanced effect with the combination did not extend to TxB2 or cyclic AMP production. Propranolol, PGI2, and the combination inhibited TxB2 synthesis to a similar extent, and PGI2 produced a similar increase in cyclic AMP in the presence and absence of propranolol. These findings indicate that propranolol and PGI2 inhibit platelet aggregation through cyclic AMP-independent and dependent mechanisms, respectively. While propranolol does not alter the synthesis of PGI2, it enhances the inhibition of aggregation by PGI2, and this may contribute to its antiplatelet effect. PMID- 2986879 TI - Angiotensin-converting enzyme: confusion about activity units. PMID- 2986878 TI - Immunologic behavior of lymphocytes in experimental viral myocarditis: significance of T lymphocytes in the severity of myocarditis and silent myocarditis in BALB/c-nu/nu mice. AB - To clarify the immune mechanism in myocarditis, we examined by immunofluorescence techniques the serial changes in percentages of T and B lymphocytes in the heart, spleen, and peripheral blood of DBA/2 mice inoculated with encephalomyocarditis (EMC) virus (experiment I). B cells were demonstrated by staining with fluorescein isothiocyanate (FITC)-labeled rabbit antimouse immunoglobulin (Ig). T cells were demonstrated with rat anti-Thy1.2 monoclonal antibody plus FITC labeled antimouse Ig. There was a marked decrease in T cells in peripheral blood and a moderate decrease in the cells in the spleen on day 14. There were no significant changes in B cells in peripheral blood or spleen throughout the entire period and T cells accounted for approximately 80% of the cells in the myocardium on days 7 and 14. To confirm the involvement of T cells in the development of myocarditis, we also carried out studies in which BALB/c-nu/nu mice (group 1, n = 58), BALB/c-nu/+ mice (group 2, n = 54), and BALB/c-nu/nu mice injected with 5 X 10(7) spleen cells from BALB/c-nu/+ mice (group 3, n = 50) were inoculated with EMC virus (experiment II). Four mice from each of the three groups were killed on day 6 for virologic studies. In experiment II, there were no significant differences in the incidence of myocarditis among the three groups. Virus titrations of the heart and serum neutralizing antibody titers did not show any significant differences between the three groups on days 6 and 16.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986880 TI - Applications of recombinant DNA to pathologic diagnosis. AB - Recombinant DNA techniques are contributing to the understanding of the pathogeneses of genetic, neoplastic, and viral diseases, and are used in the diagnosis of certain genetic and viral diseases. Such techniques will have wider application in the future and will play an increasing role in the clinical laboratory. The technology of this field rests upon the cleavage of DNA by certain enzymes, restriction endonucleases, and upon the ability to locate specific sequences of nucleotides in a cleaved DNA sample by using known fragments of DNA ("probes") labeled with radioisotopes or biotin. To produce useful probes, one "clones" multiple copies of the same DNA fragment in bacteria. The use of DNA probes in the clinical laboratory is valuable in antenatal diagnosis, genetic counseling, and post-natal diagnosis of genetic diseases, especially hematologic diseases and inborn errors of metabolism. DNA probes can also be used to detect viral genetic material in clinical specimens. PMID- 2986881 TI - Development and evaluation of a radioimmunoassay for Arg8-vasopressin, after extraction with Sep-Pak C18. AB - We describe a specific double-antibody radioimmunoassay for measuring arginine vasopressin (AVP) in human plasma. Antisera of high avidity were obtained from rabbits that had been injected with AVP coupled to bovine thyroglobulin. The antibody reacts with both the tripeptide tail and the pentapeptide ring of the molecule, thereby eliminating cross reaction with oxytocin. Synthetic AVP was labeled with 125I by a modification of the Chloramine-T technique. The specific activity of the labeled hormone was 29 MBq/micrograms of AVP, as estimated by self-displacement analysis. The assay involves Sep-Pak C18 extraction of acidified (pH 4) plasma. Recovery of [3H]AVP added to plasma averaged 86.6 (SD 6.1)% (n = 14). Dilution curves for plasma showed linearity of response with concentration. The overall sensitivity was 0.3 ng/L when 2-mL plasma samples were extracted. The intra-assay CV was 7.8% at 4.8 ng/L (n = 12) and the interassay CV was 12.3% (n = 16) and 6.3% (n = 14) at 2.7 and 4.1 ng/L concentrations, respectively. PMID- 2986882 TI - Cardiovascular effects of a renin inhibitor in relation to posture in nonhuman primates. AB - Orthostatic cardiovascular reflexes were evaluated in conscious cynomolgus monkeys during interruption of the renin-angiotensin system with the renin inhibitor: RIP (Pro-His-Pro-Phe-His-Phe-Phe-Val-Tyr-Lys-OH). RIP was synthesized via solid phase techniques and purified to homogeneity. In vitro studies indicated that it exhibited classical competitive inhibition of renin with a KI of 2.3 microM. In vivo, RIP at 2 mg/kg per min inhibited renin and angiotensin I pressor responses indicating that it was not a specific renin inhibitor at this dose. However, in spite of the nonspecificity, RIP did not affect the supine blood pressure of sodium-replete monkeys, but did evoke hypotension in supine sodium depleted monkeys. RIP did not elicit significant orthostatic hypotension in either sodium-replete or sodium depleted monkeys. The cardiovascular effects of RIP described in this study appear to be due to inhibition of the renin angiotensin system. PMID- 2986883 TI - Effect of sodium chloride on opiate receptor binding in spontaneously hypertensive rats: dependence on age. AB - The effect of 100 mM sodium chloride on the binding of [3H]-naltrexone to rat brain opiate receptors in spontaneously hypertensive (SHR) and normotensive (WKY) rats was studied. The percentage increase in binding in the presence of sodium chloride did not vary with age in WKY rats. Brain homogenates from 4 week old SHR rats incubated with 100 mM sodium chloride exhibited a similar increase in binding compared to age matched WKY rats. In contrast, brain preparations from 6, 14 and 20 week old SHR rats were more sensitive to sodium chloride, and the increase in binding of [3H]-naltrexone was significantly greater in these animals than in corresponding normotensive ones. Since blood pressure is increased in SHR rats compared to WKY rats at these ages, these results suggest that elevated blood pressure may be correlated with an increase in opiate receptor sensitivity to sodium chloride. The effect of in vivo sodium chloride was examined by feeding the animals a diet containing 4% salt. This concentration of salt did not significantly alter the binding of [3H]-naltrexone to rat brain homogenates prepared from 8 week old SHR rats. These results suggest that higher levels of sodium chloride and longer exposure to the diet may be required to observe the salt sensitivity produced by 100 mM salt in the in vitro radioreceptor assay. PMID- 2986884 TI - DNA polymorphism of the RC8 probe on the X-chromosome. Identification of a new DNA variant with the TaqI enzyme. AB - Restriction fragment length polymorphisms detectable with the RC8 probe, a probe for an area located on the short arm of the X-chromosome, and loosely linked to the locus for Duchenne muscular dystrophy, have been studied in a Norwegian population. With the TaqI enzyme three variants were observed. The gene frequencies of the previously detected variants were 0.867 and 0.082, respectively, and the frequency of a new variant was 0.051. Family studies confirmed Mendelian inheritance of the variants. PMID- 2986885 TI - Surface markers, 5'-nucleotidase activity, and in vitro functions of lymphocytes from patients with primary humoral immunodeficiency. AB - Immunologic surface markers, 5'-nucleotidase activity, and in vitro cell functions were determined on peripheral blood mononuclear cells from 20 patients with primary humoral immunodeficiencies. Imbalances of T-cell subsets were a frequent finding and often associated with a deficiency of helper T lymphocytes, predominantly in common variable immunodeficiency. The magnitude of the blastogenic response to mitogens was subnormal in the majority of the samples and showed no correlations to the proportions of helper, suppressor T cells or monocytes in the cultures. Moderate to marked suppression of the pokeweed mitogen driven allogenic B-cell maturation was mostly mediated by T lymphocytes but modified by monocytes in some patients. In identical twins, T lymphocytes selectively suppressed IgA production. Both in sex-linked agammaglobulinemia and common variable immunodeficiency the activity of 5'-nucleotidase was significantly lower than in controls. This deficiency was mediated by T lymphocytes and showed no correlations to imbalances of T-cell subpopulations or to alterations of lymphocyte functions. PMID- 2986886 TI - Carbon-PGLA prostheses for ligament reconstruction. Experimental basis and short term results in man. AB - The efficiency of a pure carbon prosthesis to reconstruct intra-articular ligamentous loss of substance was evaluated on 14 sheep with an 18-month follow up study of recent and old ACL experimental injuries. Functional results showed a correlation between a tightened implant, the stability of the knee, and the absence of severe arthritis. The rupture strength of the newly formed ligament amounts to about 300 Newtons (N). That of the normal ACL amounts to 250-550 N. The structure gives a viscoelastic behavior to the prosthesis. This tissue was made of collagen fibers surrounding the carbon fibers and running generally in the direction of the ligament. In the osseous tunnels, the newly formed lamellar bone invaded and surrounded the implant. Carbon fiber fragmentation occurred, and fibers were found in the synovia and in the homolateral, inguinal, and paraaortic lymph nodes. No degeneration, necrosis of tissue, or cellular toxicity was found. On these bases, the carbon prosthesis was modified by adding a resorbable copolymer of polyglycollic (PGA) and polylactic acid (PLA) around the fibers and a resorbable sheath of the same polymer. This carbon-PGA/PLA prosthesis was used in 23 patients with a three- to 12-month follow-up period. Patients were evaluated by clinical tests and by functional control of the ligament. The indications for prosthetic replacement were: recent mop-end tears of the ACL, cruciate ligament reconstruction in chronic knee instability, and wide rotator cuff loss of substance. Five complications occurred because of hematomas and/or sepsis and prosthesis breakage. Eighteen of 23 patients had good clinical results with good function in five. The conclusions are that PGA/PLA produces a clean and flexible ligament, thus eliminating carbon fiber articular deposits and allowing a normal function. Inflammatory postoperative reactions seem to be more frequent than with other procedures and call for operative and clinical care in order to eliminate hematoma and sepsis. The efficiency of a rehabitable carbon-PGA/PLA prosthesis in intra-articular ligamentous defects in man will be confirmed only by controlled long-term clinical observations. PMID- 2986887 TI - [Immunocytochemistry of Pick's argentophilic bodies: evidence for the involvement of high molecular weight microtubule-associated proteins (HMWP) before the appearance of tubulin]. PMID- 2986888 TI - Localization of enlarged parathyroid glands by thallium-201 and technetium-99m subtraction imaging. Gland mass and parathormone levels in primary hyperparathyroidism. AB - Twenty-two patients, all with surgically proven primary hyperparathyroidism, were studied by TI-201 thallous chloride and Tc-99m pertechnetate subtraction imaging. Fifteen parathyroid adenomata and one hyperplastic gland between 0.33 and 14.8 g were correctly localized in 16 patients. Two adenomata and seven hyperplastic or histologically normal parathyroids between 0.1 g and 1.4 g in seven patients were not localized. One patient had a correctly localized 13.0-g adenoma with a nonlocalized 0.3 g hyperplastic parathyroid gland and there were two false positive localizations. Sensitivity was 64% (glands), and 73% (patients). There was only fair correlation with parathormone (PTH) levels, but these were elevated in all but four of the patients with correctly localized parathyroids. The authors conclude that the imaging procedure is useful but its sensitivity is limited by difficulty in localizing correctly small glands, particularly those of less than 0.5 g, which comprised 29% of those excised. PMID- 2986889 TI - Giant parathyroid adenoma demonstrated by dual tracer parathyroid imaging. AB - Noninvasive diagnostic imaging modalities have played an increasingly important role in preoperative localization of parathyroid lesions. The radionuclide method using TI-201 and Tc-99m subtraction imaging is quite successful in detecting such lesions, with a reported sensitivity of 92%. A case of surgically proven primary hyperparathyroidism, caused by an unusually large parathyroid adenoma and detected by the dual tracer method, is presented. PMID- 2986890 TI - Parathyroid adenomas and hyperplasia. Dual radionuclide scintigraphy and bone densitometry studies. AB - Dual radionuclide imaging of the neck (Tc-99m pertechnetate and TI-201 thallous chloride) was performed in 22 patients with chemical evidence of hyperparathyroidism (elevated blood calcium and parathormone [PTH] levels). Of these, 19 of 22 had localization of a TI-201 "excess" area on radiothallium radiotechnetium subtraction images. In 13 patients who have had operative confirmation at this writing, the dual radionuclide imaging was positive in 12 (12/13 = 92.3%). One patient had two parathyroid adenomas, both of which were seen on the images; thus, overall detection was 13 out of 14 or 92.8%. The parathyroid adenomas visualized weighed from 0.06 to 3.0 g; the one not detected weighed 0.25 g. In three patients with parathyroid hyperplasia secondary to renal disease, the subtraction imaging detected eight of 12 glands (66.7%). The forearm bone mineral content and bone density were determined in eight patients with parathyroid adenomas; results were abnormally low in five of these eight. One of the three patients with secondary hyperparathyroidism had an abnormally low radial bone mass. The combination of dual radionuclide imaging and radial bone mass determination may present a useful approach in both localizing abnormal parathyroid tissue and in examining its functional consequences. PMID- 2986891 TI - Unusual scintigraphic presentation of a shifting Meckel's diverticulum. AB - A Tc-99m red blood cell (RBC) study in a child with acute gastrointestinal bleeding of unknown origin suggested Meckel's diverticulum because of free pertechnetate uptake in the ectopic gastric mucosa. This finding has not been reported in the literature. A scan with Tc-99m pertechnetate then demonstrated a focal area of uptake with an unusual pattern of abdominal shifting consistent with a Meckel's diverticulum. A large Meckel's diverticulum containing gastric mucosa was found at surgery. A shifting pattern of focal uptake on a meckelogram study should be included among the diagnostic criteria for Meckel's diverticulum. PMID- 2986892 TI - A case of synovial sarcoma with bone metastasis identified by bone marrow scintigraphy. AB - In a patient with synovial sarcoma, routine bone survey showed no abnormality, while bone marrow scintigraphy with Tc-99m sulfur colloid revealed a defect in the fifth lumbar vertebra. At surgery, tumorous invasion was noted in the fifth lumbar vertebra and the surrounding tissues. It was suggested that the bone marrow scintigraphy was particularly useful in the detection of tumorous invasion into the bone marrow at the early stage before the destruction of skeletal tissue. PMID- 2986893 TI - Medial splenic tubercle. Potential radionuclide ventriculography pitfall. AB - Blood pool activity in a medial splenic tubercle produced an apparent apical wall motion abnormality during radionuclide ventriculography. PMID- 2986894 TI - Dual isotope subtraction parathyroid scintigraphy in the preoperative evaluation of suspected hyperparathyroidism. AB - Ninety-five percent of primary hyperparathyroidism is caused by either a solitary hyperfunctioning adenoma or chief cell hyperplasia. While there is a great deal of variation in the location of the parathyroid glands, 90-95% of all abnormal parathyroid tissue will be identified at initial exploration regardless of the preoperative localization procedure. Adenomas not identified at initial exploration are likely to be ectopic. Reexploration is a difficult and time consuming procedure. The challenge, therefore, is to develop a reliable, noninvasive test that will locate aberrant parathyroid tissue preoperatively with the goal of simplifying the surgical procedure and reducing the incidence of reexploration. A radiopharmaceutical which localizes selectively in parathyroid tissue would be ideal. Currently, no single radiopharmaceutical is adequate. Recently, dual isotope subtraction scintigraphy using T1-201 chloride and Tc-99m pertechnetate has yielded promising results. The authors have investigated this procedure in 42 patients, 25 of whom to date have had surgical exploration. Of the 25 adenomas or focal hyperplasia sites found at surgery, 23 (92%) were correctly located preoperatively. There were two false-negative sites and five false-positive sites. The purpose of this paper is not only to describe the methods used and results obtained but also to discuss the merits and limitations of the technique in comparison with other imaging modalities so that its role, in today's climate of cost containment, may better be defined. PMID- 2986895 TI - The role of human papilloma and other viruses. PMID- 2986896 TI - Problems in cytological screening from condylomatous lesions of the cervix. PMID- 2986897 TI - The pathology of cervical cancer. AB - The preinvasive phase of squamous cell carcinoma of the cervix is a continuous spectrum of abnormal epithelium, which, for convenience of classification and as a guide to management, is customarily subdivided into three grades. The histological diagnosis of CIN, as well as the distinction between the grades, depends on a combination of features embracing aspects of differentiation, nuclear changes and mitotic activity. Grading of CIN is subjective. Generally, a minor degree of CIN would be expected to progress to a more severe form if not treated, but this progression does not seem to be inevitable; the more severe a CIN is at the time of diagnosis, the more likely it is that it will progress, both to a more severe degree of CIN and, eventually, to invasive carcinoma. Conversely, the more minor the degree of CIN at diagnosis, the more likely it is that it will regress. True figures are not available for the rate of progression from CIN to invasive carcinoma; it is sufficient to accept that the risk of progression probably occurs in a significant proportion of cases, if not the majority. Preclinical invasive carcinoma is divided into microinvasive carcinoma and occult invasive (Stage Ib) carcinoma. The definitions of these lesions have not yet been satisfactorily established; the term microinvasive carcinoma should define the maximum size of tumour which has virtually no metastatic potential and so may be treated in a conservative fashion. Invasive squamous cell carcinoma is classified histologically according to the cell type and the degree of differentiation, although it is debatable whether the cell type has any correlation with prognosis. Adenocarcinomas make up 5-10% of cervical cancers and a variety of histological types have been recognized. Adenocarcinoma in situ is being diagnosed with increasing frequency, often in association with squamous CIN. It seems apparent that AIS is a precursor of adenocarcinoma, but little is known about its natural history. PMID- 2986898 TI - Bone grafting in mandibular alveolar ridge augmentation. Current status. PMID- 2986900 TI - Lymphocyte phenotype changes in congenital and acquired perinatal cytomegalovirus infections. AB - The recognition that the multiple functions of T-lymphocytes are accomplished by separate subpopulations of cells that can be delineated both functionally and by antisera directed against specific cell surface markers is one major advance in immunology. Our study was directed at enumerating lymphocyte subsets in cytomegalovirus (CMV) perinatal infections with monoclonal antibodies and cell flow cytometry. Eight patients ranging in age from 1 week to 2 1/2 months were studied. Except for one term neonate, all of the infants were less than 36 weeks of gestational age, with a mean of 31.5 weeks (range 27-36 weeks) at birth. The mean birth weight of the premature infants was 1,211 gm (range 870-2420 gm). Five patients were 3 weeks old when studied and were classified as congenital CMV by viral isolation. Three were older than 2 months and were classified as acquired CMV. One of the acquired CMV infants was followed from birth to 6 months of age and had normal lymphocyte numbers at 2 weeks with alterations appearing at 8 weeks. The results of these studies indicated that in congenital CMV there is a normal to high helper/suppressor ratio. On the contrary, in acquired CMV this ratio is profoundly depressed because of significant increase in suppressor T lymphocytes with borderline normal helper T-lymphocytes. In the latter group, the ratio increased with time after the acute infection episode. Our results agree with reported data indicating that in adults with CMV there is a depression of the helper/suppressor ratio during the acute stage with a return toward normal values during convalescence. PMID- 2986899 TI - Subgingival application of a hydrogen peroxide/baking soda mixture with a toothpick. Periodontal effects. PMID- 2986901 TI - Supine and sitting maximal workload difference and response to long-term enalapril therapy in congestive heart failure. AB - Maximal workloads in supine and sitting bicycle stress tests were compared in 12 patients with congestive heart failure during long-term therapy with the vasodilator enalapril. The response to the therapy was observed with the supine and sitting bicycle stress tests and gated equilibrium ventriculography. The maximal workload was 68 W in the supine position and 84 W in the sitting position; the mean difference was 15 W. The difference was highly significant (P less than 0.001). Both the supine and sitting workloads were significantly higher (P less than 0.02 and P less than 0.05) in the enalapril group than in the placebo group. The findings in the gated equilibrium ventriculography were not significantly different between the two groups. PMID- 2986902 TI - Patterns of perfusion and ventilation of the lungs in patients with small cell lung cancer before and after combination chemotherapy. PMID- 2986903 TI - Routine typing of herpes simplex virus isolates. PMID- 2986904 TI - Glucose-1,6-P2 synthesis, phosphoglucomutase and phosphoribomutase correlate with glucose-1,6-P2 concentration in mammals red blood cells. AB - Glucose 1,6-biphosphate (G1,6P2) was measured in human, pig, cow, rabbit, rat and sheep red blood cells. Mean values are variable among the species and range from 33 to 122 nmol/ml RBC for pig and rabbit erythrocytes, respectively. The activities of G1,6P2 synthase, phosphoglucomutase (PGM) and phosphoribomutase (PRM) have also been assayed in red cell haemolysates of the same species. The correlations between the biphosphate content and the occurrence of the three enzymatic activities have been studied in order to gain an insight into the regulation of the G1,6P2 turnover in mammalian erythrocytes. PMID- 2986905 TI - Endogenous digitalis-like factors revisited 1981-1985. A review. PMID- 2986906 TI - Neural modeling and model identification. AB - The complexity of the nervous system poses challenging experimental and theoretical problems to the investigator. While experimental studies have provided a broad understanding of the physiological and anatomical role of neural elements, a comprehensive description of how the nervous system acquires and processes information is still lacking. Neural modeling addresses itself to the quantitative interpretation of neurophysiological experiments, to the assimilation of diverse experimental results into unified theories, and to the investigation of mechanisms of information-processing units, codes, and networks and their relation to capabilities of living systems. This review will emphasize neural models which can be tested by physiological or psychophysical experiments and will include the identification of model parameters based on experimental measurements. An overview of neural firing models, neural interaction models, neural variability and coding, design of input-output experiments for model identification, and description of small network interactions through multiunit and gross potential recording will be included. PMID- 2986907 TI - Variable adrenocortical function in acute medical illness. AB - Acute medical illness may produce dramatic changes in endocrine function. Although cortisol levels rise in acute medical illness, changes in adrenocortical function and reserve have not been well documented in medical ICU patients. We evaluated plasma ACTH and cortisol levels, and cortisol response to intravenous ACTH in 40 acutely ill patients and 20 anxious but nonacutely ill controls. A wide range of plasma cortisol values (212 to 8430 nmol/L) was observed. More severely ill patients did not necessarily have higher plasma cortisol values. Patients who survived hospitalization had lower mean initial and post-ACTH cortisol levels than patients who succumbed. ACTH levels varied widely and correlated poorly with levels of plasma cortisol. There was no evidence of occult adrenocortical insufficiency. We conclude that plasma cortisol elevations are common in acute medical illness. PMID- 2986908 TI - DNA topoisomerases: enzymes that control DNA conformation. PMID- 2986909 TI - Granular cell tumor: a great masquerader. AB - Granular cell tumor usually presents as an asymptomatic mass in the skin or subcutaneous tissue, yet it is rarely considered in the clinician's differential diagnosis of such masses. We present a case of granular cell tumor located on an extremity and discuss its characteristic histopathologic findings and aspects of its origin and management. PMID- 2986910 TI - Papulovesicular acrolocalized syndrome. AB - A 1-year-old boy with a cutaneous eruption compatible with papulovesicular acrolocalized syndrome is described. This unusual but benign entity of early childhood can be differentiated both by its clinical presentation and appropriate laboratory testing from the two morphologically similar eruptions: Gianotti Crosti syndrome and infection with Coxsackie virus A-16. PMID- 2986911 TI - Zeolite exposure and associated pneumoconiosis. AB - Naturally occurring zeolite minerals are aluminum silicates widespread in the earth's crust. Several of these minerals have fibrous forms and have been implicated as a possible cause of benign and malignant diseases of the lung and pleura in Turkey. This report describes a patient, living in an area of Nevada rich in zeolites, who presented with idiopathic pleural thickening and pulmonary fibrosis associated with extensive pulmonary deposition of zeolites. PMID- 2986912 TI - Serum angiotensin-converting enzyme in coal worker's pneumoconiosis. PMID- 2986913 TI - [Liver resection. Indications and results]. PMID- 2986914 TI - [Glomus tumor as a cause of coccygodynia]. AB - Painful glomus tumors near the coccyx have not yet been described before. Three patients with coccygodynia were treated by excision of the glomus coccygeum. All of them were free of pain afterwards. Two glomus tumors were found. The tumors of the glomus coccygeum are compared with the well-known glomus tumors "Masson". PMID- 2986916 TI - [Adenoid cystic carcinoma of the salivary gland: a clinicopathologic study of 225 cases]. PMID- 2986917 TI - [Hepatic lobectomy of 84 primary liver carcinomas]. PMID- 2986915 TI - [The Roentgen features of pulmonary metastases of primary carcinoma of the liver (report of 102 cases)]. PMID- 2986919 TI - [Bronchofiberscopy and pathologic findings in small cell lung cancer]. PMID- 2986918 TI - [Humoral leukocyte adherence inhibition test in lung cancer]. PMID- 2986920 TI - [Clinical analysis of 28 cases of bronchial adenoma]. PMID- 2986921 TI - Exacerbation of diarrhea after iodinated contrast agents in a patient with VIPoma. PMID- 2986922 TI - Sarcomas arising after radiotherapy for peptic ulcer disease. AB - Therapeutic gastric irradiation has been used to reduce peptic juice secretion in patients with peptic ulcer disease. Between 1937 and 1968 a total of 2049 patients received such therapy at the University of Chicago. Three of these patients are known to have developed sarcomas in the field of radiation. Two gastric leiomyosarcomas of the stomach were diagnosed 26 and 14 years after treatment and a malignant fibrous histiocytoma of the anterior chest wall was removed six years after gastric irradiation. Of 743 peptic ulcer patients treated without irradiation and constituted as a control group for the study of therapeutic gastric radiation, none is known to have developed sarcoma. As the incidence of sarcoma in these patient groups is known only from the tumor registry of the University of Chicago, other cases of sarcoma may exist in the groups. While an increased incidence of sarcoma has not been proven to occur in patients who received therapeutic gastric irradiation for peptic ulcer disease, the possibility of such a risk should be borne in mind by physicians caring for such patients. PMID- 2986923 TI - Fiber and diabetes. PMID- 2986924 TI - Mathematical model to obtain cholesterol, P/S ratio, and fiber daily intake. PMID- 2986925 TI - [Nature of the electrogenic phases of the bacteriorhodopsin photocycle and the localization of retinal]. PMID- 2986927 TI - [Cooperative killing of yeast cells by ozone-induced dissipation of the transmembrane proton gradient]. PMID- 2986926 TI - [Changes in the paramagnetic complexes of the blood and liver of animals exposed to nitroglycerin]. PMID- 2986928 TI - [Oligonucleotide-directed mutagenesis. The introduction of point mutations (transitions, deletions, insertions) in the beta-galactosidase gene]. PMID- 2986929 TI - Nalbuphine. AB - Nalbuphine is a potent analgesic with a low side effect and dependence profile in animals and man. Nalbuphine is distinguished from other agonist/antagonist analgesics in having greater antagonist activity and fewer behavioral effects at analgesic doses than pentazocine, butorphanol or buprenorphine. At equi-analgesic doses, nalbuphine is quantitatively similar to nalorphine in regard to its large ratio of antagonist to analgetic activity. Clinical studies have confirmed this balance of strong antagonist to analgesic activity. Nalbuphine has been shown to effectively antagonize the respiratory depressant activity of narcotic analgesics while concomitantly adding to their analgetic responses. Unlike nalorphine or pentazocine, nalbuphine produces few overt behavioral or autonomic effects in animals at doses over 300 times its analgesic range. These findings are confirmed by clinical results which show that nalbuphine produces few psychotomimetic effects, even at elevated dose levels, in contrast to nalorphine or pentazocine. Nalbuphine produces limited respiratory depression in animals and in man. Significant cardiovascular effects have not been found. Nalbuphine was found to produce significantly less inhibition of gastrointestinal activity than any of the clinically useful narcotic or agonist/antagonist analgesics tested in animals. Nalbuphine's analgetic effects are reversed by naloxone doses similar to those which reverse nalorphine's agonist effects. Results in this and other tests suggest that nalbuphine is primarily a kappa-agonist/mu-antagonist analgesic. Unlike pentazocine or buprenorphine, nalbuphine does not suppress the narcotic abstinence syndrome in partly-withdrawn morphine-dependent animals or man. Rather, due to nalbuphine's strong antagonist activity, analgesic-range doses of nalbuphine severely exacerbate the withdrawal syndrome in partly-withdrawn mice, monkeys and humans. Nalbuphine also precipitates a strong abstinence response in non-withdrawn morphine-dependent animals and man. In post-addict humans, analgesic-range doses of nalbuphine are perceived as minimally morphine-like, but higher doses are judged to be progressively more nalorphine-like (i.e. dysphoric), which further limits nalbuphine's abuse potential in drug-seeking individuals. Primary dependence studies have demonstrated that physical dependence is possible at high dose levels that produce marked side effects. Other studies show that dependence is unlikely to be of significance within nalbuphine's usual analgesic range. Six-month studies in patients with chronic pain have confirmed that analgesic tolerance or physical dependence is uncommon.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986930 TI - Buprenorphine. AB - Buprenorphine is a mixed agonist-antagonist with high affinity at both mu and kappa opiate receptors. Its pharmacological profile is determined primarily by partial agonism at mu-receptors and unusually slow kinetics at these receptors. Its intrinsic activity is such that in nearly all clinical situations it is as effective an analgesic as morphine with considerably longer duration and much more favourable acute safety. In long-term dosing studies in rodents and primates buprenorphine did not produce the manifestations of physical dependence when treatment was stopped. In self-administration studies in the same species only limited levels of reinforcing efficacy were demonstrated when compared with the opiates. In human former opiate addicts the limited potential of buprenorphine to produce psychological dependence was confirmed as was the favourable physical dependence profile. Some misuse of buprenorphine has been reported in 3 of the 29 countries in which buprenorphine is marketed despite its wide clinical acceptance, particularly as the sublingual formulation. PMID- 2986931 TI - Picenadol. AB - Picenadol is a unique opioid mixed agonist-antagonist analgesic currently under clinical evaluation. Structurally, picenadol is a 4-phenylpiperidine derivative and a racemic mixture whose mixed agonist-antagonist properties are a consequence of the d-isomer being a potent opiate agonist, whereas the l-isomer is an opioid antagonist. In the mouse writhing and rat tail heat tests, the analgesic potency of picenadol is estimated to be 1/3 that of morphine. Picenadol itself has weak antagonist activity, whereas the antagonist potency of the l-isomer is approx. 1/10 that of nalorphine. Evaluation of picenadol's affinity for opioid receptors reveals that picenadol, unlike other mixed agonist-antagonists has high affinity for both the mu and delta receptors but a markedly lower affinity for the kappa receptor. Extensive pharmacological investigations show picenadol to have a low potential to produce opiate-like side effects, including a low liability for abuse and physical dependence. PMID- 2986932 TI - [Effect of an introductory dose of etomidate, methohexital and midazolam on adrenal cortex function before and after ACTH-stimulation]. AB - The adrenocortical function after induction of anaesthesia using either etomidate, methohexital or midazolam was examined in 30 patients undergoing open heart surgery (ACVB). Midazolam and methohexital did not suppress the typical stress response assessed from the magnitude of the ACTH, cortisol and aldosterone levels. The ACTH-test was also normal. In contrast, induction of anaesthesia using etomidate led to a progressive fall in cortisol and aldosterone accompanied by an increase in ACTH. The ACTH-test resulted in only a minor increase in adrenocortical hormones. Thus an induction dose of etomidate, in contrast to methohexital and midazolam, markedly alters the normal function of the adrenal cortex. This side-effect of etomidate lasts for at least 4 hours but is no longer detectable after 24 hours. PMID- 2986933 TI - [Cytomegalovirus infections in AIDS]. PMID- 2986934 TI - [Transitory oligoclonal paraproteinemia with virus infection and malignant lymphatic disease]. AB - Oligoclonal paraproteinaemia occurred in two patients with malignant lymphocytic disease (highly malignant non-Hodgkin lymphoma, B-cell type of acute lymphocytic leukaemia), in one case during a cytomegalovirus infection and in the other during an infectious mononucleosis. At that time both patients were in complete remission. Paraproteinaemia in the first case disappeared within a year where the transformation from an initially four-banded paraproteinaemia (2 IgM-lambda and 2 IgG-lambda) into a three-banded paraproteinaemia (IgG-lambda) could be observed. In the second patient the concentration of the paraprotein decreased considerably. Because both patients were no longer under cytostatic treatment after manifestation of the paraproteinaemia, and were in complete remission during the whole of the observation period (4 years and 1 year), a direct relationship with the primary disease is unlikely. The transitory paraproteinaemia appears to be rather the result of an acquired defect of the immune system. PMID- 2986935 TI - [Pseudohypertriglyceridemia in glycerokinase deficiency]. AB - Falsely high serum triglyceride concentrations (410-850 mg/dl) were measured in four members of a family of five from Franconia in Germany. The cause was hyperglyceridaemia on the basis of glycerol kinase deficiency. None had any symptoms and no other metabolic anomaly was demonstrated. The possibility of glycerol kinase deficiency should be considered in any case of elevated serum triglyceride concentration but with clear serum, normal lipid electrophoresis and lack of response to lipid-lowering measures. PMID- 2986936 TI - [Global pituitary stimulation test with releasing hormones]. AB - The endocrine function of the thyroid and gonads has for long been investigated using the corresponding releasing hormones (TRH- and LHRH-test, respectively). The adrenal cortex has, up to now, been stimulated using insulin-induced hypoglycaemia or lysine-vasopressin and growth hormone stimulated using arginine. New diagnostic possibilities have arisen with the isolation of the corresponding releasing-hormones, CRF and GRF, and with the availability of these too for clinical use. Using the four above mentioned releasing-hormones in a global pituitary-stimulation-test, the secretion of ACTH, cortisol, STH, TSH, LH, FSH and prolactin hormones can now be examined together. PMID- 2986938 TI - Injuries to the major nerves of the hand. AB - This article outlines methods that can be used in the emergency department or in the physician's office to establish the presence of injury to a peripheral nerve in the upper limb. This includes motor and sensory tests that require equipment no more sophisticated than a paper clip but that give accurate, reproducible, clinically useful information. PMID- 2986937 TI - Amiodarone. Biochemical aspects and haemodynamic effects. AB - The mechanisms underlying the non-competitive beta-antagonistic properties of amiodarone were investigated, and the haemodynamic responses to exercise following the administration of oral amiodarone or intravenous propranolol were compared in dogs with a healed myocardial infarction submitted to a graded treadmill exercise. In radioligand binding studies, amiodarone, up to 10 mumol/L did not compete with 125I-iodocyanopindolol for binding to rat heart beta adrenoceptors. Exposure of cardiac membranes to greater concentrations of amiodarone induced a significant decrease in the number of beta-adrenoceptors without affecting their affinity for 125I-iodocyanopindolol. Similar results were observed ex vivo, in rats after single or multiple dose administration. When added in vitro to rat heart membranes, amiodarone non-competitively inhibited the activation of adenylate cyclase by isoprenaline, glucagon and secretin. Stimulation of adenylate cyclase by those agents which act at more internal sites in the sarcolemmal membrane such as GppNHp, sodium fluoride or forskolin, was much less affected by amiodarone. In dogs performing at a submaximal work level, amiodarone significantly reduced heart rate and tended to increase coronary flow and to reduce left ventricular end-diastolic pressure, but did not affect left ventricular dP/dt. During submaximal exercise, propranolol had similar effects on heart rate, but dramatically reduced myocardial contractility. PMID- 2986939 TI - Corticotropin-releasing factor binding to peripheral tissue and activation of the adenylate cyclase-adenosine 3',5'-monophosphate system. AB - Specific binding sites for rat corticotropin-releasing factor (rCRF) are present in rat adrenal medulla, ventral prostate, spleen, liver, kidney, and testis and bovine chromaffin cells in culture. Maximal binding of [125I]rCRF occurred within 25 min at 4 C and was saturable. Scatchard analysis of rCRF binding to rat adrenal membranes and bovine chromaffin cells revealed the existence of two classes of binding sites. One class had a relatively higher apparent affinity and lower number of binding sites, whereas the other class had a relatively lower affinity and higher number of binding sites. CRF induced a dose-related increase in rat adrenal membrane adenylate cyclase activity and cAMP levels in bovine chromaffin cells. Nanomolar concentrations of rCRF maximally stimulated adenylate cyclase activity in rat adrenal membranes and maximally increased cAMP levels in bovine chromaffin cells to 86% and 130% above control values, respectively. The demonstration of specific CRF-binding sites in a variety of peripheral tissues and the finding that activation of specific CRF-binding sites in adrenal tissue stimulates the adenylate cyclase-cAMP system suggest that CRF may have an important regulatory role in various peripheral tissues. PMID- 2986940 TI - Atypical synergistic alpha 1- and beta-adrenergic regulation of adenosine 3',5' monophosphate and guanosine 3',5'-monophosphate in rat pinealocytes. AB - The adrenergic control of cAMP and 3',5'-cyclic GMP (cGMP) in dispersed adult rat pinealocytes was investigated. Norepinephrine treatment increased cAMP and cGMP content 60- and 400-fold, respectively; both alpha- and beta-adrenoceptors had to be activated for these responses to occur. Beta-Adrenergic stimulation alone produced only about 6- and 2-fold increase in cAMP and cGMP content, respectively. Alpha-Adrenergic stimulation, which alone had no effect on either cyclic nucleotide concentration, markedly amplified the beta-adrenergic stimulation of both cAMP and cGMP. The relative potency of alpha-adrenergic agonists and antagonists indicates the alpha 1-subclass of adrenoceptors is involved. A role of alpha 1-adrenoceptors in the control of pineal cAMP is consistent with published evidence of the presence of alpha 1-adrenoceptors on pinealocytes and their role in the regulation of N-acetyltransferase activity and melatonin production. PMID- 2986941 TI - Modulation by cortisol of adrenocorticotropin-induced activation of adrenal function in fetal sheep. AB - We examined the hypothesis that cortisol (F) modulates the activation of adrenal function induced by treating fetal sheep in vivo with pulsatile ACTH (P-ACTH). Chronically catheterized sheep fetuses were infused in utero for 100 h between day 127 and day 131 of pregnancy with P-ACTH; P-ACTH plus metopirone; P-ACTH plus metopirone plus F; P-ACTH plus metopirone plus dexamethasone, or saline (controls). After 100 h, basal and ACTH-stimulated output of 11-desoxycortisol (S), F, and progesterone from collagenase-dispersed fetal adrenal cells was measured. Adrenal cells from fetuses treated with P-ACTH in vivo had significantly greater basal and stimulated (delta) outputs of F and S in vitro than controls. These effects were attenuated in fetuses pretreated with P-ACTH plus metopirone. Concurrent in vivo treatment with ACTH plus metopirone plus F restored basal and delta outputs of F and S to values that were not significantly different from those after P-ACTH alone. In vivo treatment with dexamethasone in addition to P-ACTH plus metopirone significantly raised basal outputs of F and S, but the cells were unresponsive to ACTH in vitro. Basal output of progesterone was significantly greater after in vivo P-ACTH plus metopirone plus dexamethasone, but no treatment raised delta progesterone output over controls. These results support a role for glucocorticoids in modulating ACTH-induced activation of adrenal function in late gestation fetal sheep. PMID- 2986942 TI - Stress-induced suppression of testicular function in the wild baboon: role of glucocorticoids. AB - In an ongoing study of endocrine function in wild olive baboons living freely in Kenya, sustained social stress was associated with suppressed testosterone (T) concentrations in males. In the present report, the acute stressor of rapid capture and immobilization caused profound and rapid suppression of T concentrations in these individuals. Elevation of cortisol concentrations preceded, and was at least partially responsible for, the declining T concentrations, as dexamethasone (DEX) administration produced a similar suppression. DEX inhibited T secretion, but did not alter its clearance. The testes appeared to be the principal site of this inhibition; DEX did not alter LHRH-induced pituitary secretion of LH, somewhat attenuated LH bioactivity, but caused a complete suppression of LH-induced testicular secretion of T. Considerable individual variation occurred in sensitivity to stress-induced suppression of T concentrations. Some individuals had transient elevations of T concentrations during the poststress hour, although concentrations ultimately declined significantly. These males were also least sensitive to DEX inhibition of LH-induced T secretion. These studies demonstrate acute stress-induced suppression of gonadal function in a population of primates living in their natural habitat. Furthermore, they implicate glucocorticoid actions mostly at the testes as possible underlying endocrine mechanisms for such regulation. PMID- 2986943 TI - Potassium fluxes in bovine adrenal cells during adrenocorticotropin stimulation. AB - Active (ouabain-sensitive) and passive (ouabain-insensitive) fluxes of 43K were measured in isolated bovine zona fasciculata/reticularis cells. ACTH inhibited passive influx in a dose-dependent manner and, at a concentration that maximally stimulated steroidogenesis (10(-8) M), also inhibited passive efflux. ACTH did not affect active potassium transport. Angiotensin II (10(-6) M) and cAMP (10(-2) M) inhibited passive influx to the same extent as did 10(-8) M ACTH. Angiotensin II (10(-6) M) also reduced active potassium uptake, although this did not appear to be related to changes in cortisol biosynthesis. Increases in cortisol synthesis by ouabain-treated cells in response to angiotensin II and ACTH were proportional to decreases in potassium influx. The addition of cortisol (1 microgram/ml) to cells slightly reduced passive potassium uptake, but not to the same extent as did 10(-8) M ACTH. Frusemide, an inhibitor of passive sodium/potassium cotransport, did not wholly abolish the effects of ACTH on potassium influx. These changes in flux are probably related to electrophysiological changes in membrane potential. Their significance in the regulation of ACTH actions has yet to be determined. PMID- 2986944 TI - Evidence for alpha 1-adrenergic stimulatory control of in vitro release of immunoreactive thyrotropin-releasing hormone from rat median eminence: in vivo corroboration. AB - The aim of this study was to investigate whether the alpha-adrenergic stimulation of TSH secretion may occur directly at the median eminence (ME) level by modulating the release of TRH. The effects of pharmacological manipulations of the two subtypes of central alpha-adrenergic receptors, alpha 1 and alpha 2, were tested on in vitro TRH release from medial basal hypothalami containing mainly the ME. Hypothalamic fragments were superfused with a modified Locke medium, and TRH was measured by RIA in samples collected every 10 min. After a preliminary period of 40 min to test TRH release during basal conditions, drug effects were checked for 20 min. Superfusion with norepinephrine (NE) (10(-10), 10(-8), 10(-6) M) induced a rapid and dose-dependent rise of TRH release; epinephrine (10(-8) M) induced an effect similar to that of NE 10(-8) M. Phentolamine (10(-7) M), an alpha-adrenergic antagonist, completely blocked the NE (10(-8) M)-induced release of TRH, which was not modified by the beta-adrenergic antagonist propranolol (10( 7) M). Neither antagonist had an effect on basal TRH release when added alone to the medium. The NE-induced release of TRH was completely suppressed by prazosin (10(-7) M), whereas yohimbine had no effect. Superfusion with clonidine (10(-9), 10(-8), 10(-7), 10(-6) M), an alpha 2-receptor agonist, did not alter basal TRH release. In contrast, phenylephrine (10(-8) and 10(-6) M), an alpha 1-receptor agonist, induced a significant (P less than 0.01) rise in TRH release. These results were corroborated in vivo in several unanesthetized rats bearing a push pull cannula previously and stereotaxically implanted into the ME. Perfusion with artificial cerebrospinal fluid containing NE (10(-7), 10(-6) M) or phenylephrine (10(-7) M) elicited a rapid rise in TRH release, within 15 min after the onset of drug perfusion. Clonidine (10(-5) M), similarly perfused for 15 min, had no effect. Our data suggest a direct stimulatory influence of catecholamines on TRH release at the ME level that is mediated through alpha 1-adrenergic receptors. PMID- 2986945 TI - Phorbol ester regulation of rat granulosa cell prostaglandin and progesterone accumulation. AB - Tumor-promoting phorbol esters are believed to affect cell functions by activating a Ca+2- and lipid-dependent protein kinase (protein kinase C). Since such protein kinases may be involved in ovarian granulosa cell metabolism, the effects of phorbol esters on prostaglandin (PG) and progesterone (P) accumulation were investigated. Cells were obtained from immature (28-29 days old) rats 48 h after injection of 20 IU PMSG and incubated for up to 5 h. A tumor-promoting phorbol ester, 12-O-tetradecanoyl-phorbol 13-acetate (TPA), at a concentration of 25 ng/ml, caused 4-fold increases in PGE and 6-keto-PGF1 alpha accumulation at 5 h. LH (10 ng/ml) caused 7- and 4-fold increases in PGE and 6-keto-PGF1 alpha accumulation, respectively. When tested in combination, the increases in PGE and 6-keto PGF1 alpha due to TPA and LH were additive. Like the effect of LH, the TPA stimulation of PG synthesis occurred after a delay of 2-3 h. By 5 h of incubation, cells exposed to TPA exhibited increased PG synthase activity in whole homogenates. TPA caused a smaller (2-fold) increase in P accumulation than was observed with LH (10-fold). When tested in combination, however, TPA decreased the P response to LH by approximately 25%. These effects of TPA on basal and LH-stimulated PG and P accumulation were very similar to the actions of GnRH. We, therefore, investigated the effect of exposure to the combination of GnRH and TPA. A GnRH agonist, [D-Ala6,des-Gly-NH2(10)] GnRH ethylamide (GnRHa; 10 ng/ml) caused a 4-fold increase in PGE accumulation. The effect of TPA on PGE accumulation was also additive to that of GnRHa. TPA, on the other hand, did not affect the 2.5-fold P response to GnRHa. Neither stimulation or inhibition of PGE or P accumulation was observed in the presence of a nontumor-promoting phorbol ester. Furthermore, TPA did not affect basal or LH-stimulated cAMP accumulation or basal or LH-stimulated protein kinase A activity. These data indicate that protein kinase C activation can influence granulosa cell PG and P accumulation. PMID- 2986946 TI - Inhibition of glucose-induced insulin release by xylazine. AB - The effects of xylazine, a sedative that is extensively used in veterinary medicine, on glucose-stimulated insulin release have been studied using isolated perifused rat pancreatic islets. Xylazine had no effect on insulin release under basal conditions. Glucose-stimulated insulin release was inhibited in a concentration-dependent manner over the range of 10(-9)-10(-6) M. Maximum inhibition even with 10(-5) M xylazine was around 80%, which suggested that xylazine is only a partial agonist at its receptor. The inhibitory effect could be completely eliminated by yohimbine, an alpha 2-adrenergic antagonist, and was unaffected by prazosin, an alpha 1-adrenergic antagonist. That xylazine is a partial agonist was verified by the fact that when present in excess, it could stimulate insulin release which had already been inhibited to 100% by epinephrine. It is concluded that xylazine is a potent partial alpha 2-adrenergic agonist and that this mechanism is responsible, at least in part, for the hyperglycemia and insulinopenia associated with its use. PMID- 2986947 TI - Tunicamycin and neuraminidase effects on luteinizing hormone (LH)-releasing hormone binding and LH release from rat pituitary cells in culture. AB - We have studied the effects of tunicamycin (TM) and neuraminidase on the binding of 125I-labeled Buserelin, a GnRH agonist, and on GnRH-stimulated LH release in cultured rat pituitary cells. Treatment with TM, an antibiotic which inhibits protein glycosylation, abolished the development of elongated cell processes without any effect on cell viability. Concomitantly, TM caused a time- and dose dependent inhibition of specific binding of Buserelin and of GnRH-stimulated LH release. The inhibition of binding was due to a decrease in the number of GnRH receptors without any significant effect on binding affinity. Protein synthesis was not affected under these experimental conditions, suggesting that the aglycosylated GnRH receptors are probably intracellularly accumulated and are not expressed on the cell surface. Treatment with neuraminidase inhibited only 50% of GnRH agonist binding and did not affect GnRH-stimulated LH release. These results indicate that the oligosaccharide portion is essential for the functional properties of the GnRH receptor. PMID- 2986948 TI - Somatomedin-C enhances induction of luteinizing hormone receptors by follicle stimulating hormone in cultured rat granulosa cells. AB - Recent studies have documented the ability of somatomedin-C (Sm-C) to synergize with FSH in the induction of rat granulosa cell progesterone biosynthesis. This direct cytodifferentiative action of Sm-C proved distinct from its established mitogenic property. To determine whether Sm-C partakes in the differentiation of granulosa cell functions other than progesterone biosynthesis, the role of Sm-C in the acquisition of LH receptors was investigated in vitro. Granulosa cells were obtained from immature, hypophysectomized diethylstilbestrol-treated rats and cultured under serum-free conditions for up to 72 h. Specific granulosa cell LH/hCG binding under basal conditions was relatively low and was not significantly affected by treatment with highly purified Sm-C (50 ng/ml). In contrast, treatment with FSH (oFSH; NIH FSH S14; 20 ng/ml) produced a 3.8-fold increase in LH/hCG binding to 3167 +/- 660 (+/- SE) cpm/10(6) cells. However, concurrent treatment with Sm-C (50 ng/ml) resulted in a 6.1-fold enhancement of the FSH effect. The ability of Sm-C to synergize with FSH in the induction of LH receptors was dose and time dependent, with an apparent median effective dose (ED50) of 6.2 +/- 0.6 (+/-SE) ng/ml, and a minimal time requirement of 24 h or less. Examination of the binding parameters of the LH/hCG receptor revealed that the Sm-C-mediated increase in LH/hCG binding was due to enhanced binding capacity, but not affinity (Kd = 4.4 X 10(-11) M). Decreasing dilutions (1:128,000-1:8,000) of a monoclonal antibody raised against Sm-C (sm 1.2), produced progressive and virtually complete immunoneutralization of the synergistic interaction between Sm-C and FSH, suggesting specificity of action. The ability of Sm-C to enhance LH receptor induction was associated with increased hCG-stimulated progesterone biosynthesis, suggesting that the newly acquired receptors are functional in nature. Our present findings indicate that nanomolar concentrations of Sm-C are capable of enhancing FSH-stimulated LH/hCG binding capacity, but not affinity, in a time- and dose-dependent fashion. These observations suggest that the direct cytodifferentiative effect of Sm-C is not limited to the induction of progesterone biosynthesis, but other key granulosa cell functions may also be subject to modulation by this regulatory peptide. PMID- 2986950 TI - Differential effects of viral infection on islet and pituitary cell lines. AB - Although reovirus infection may lead to changes in endocrine function in vivo, little is known about the precise interaction of reovirus with endocrine cells. In this study we have examined the effects of reovirus infection on two types of endocrine cells, GH4C1 cells and RINm5F cells. Both type 1 reovirus and type 3 reovirus infect the two cells lines and appear to grow equally well. Viral replication occurred within the first 24 h following infection after which viral titers remained stable for 3 days. By 48-72 h after viral infection, substantial cytopathic effects were noted in RINm5F cells infected with both type 1 and type 3 reovirus. In GH4C1 cells, type 3 reovirus was most effective in producing cell death, and type 1 reovirus was significantly less cytotoxic despite a similar viral titer. Only type 1 reovirus caused a specific inhibition of overall protein and DNA synthesis, and this occurred only in the RINm5F cells. Over the time course studied, GH4C1 cells successfully infected with type 1 reovirus demonstrated no cytopathic effects, and only minimal alterations in cellular function were noted. Intracellular insulin content and insulin secretion, a "luxury function" of the RINm5F cells, were also surprisingly well maintained in the first 48 h after viral infection. In addition, virally infected cells were able to respond to glyceraldehyde, an insulin secretagogue, although the response appeared to be somewhat blunted compared to that of control cells. These results suggest that viral infection of endocrine cells results in specific alterations that depend on the nature of the infecting virus. In addition, the cellular environment of the host cell may be an important determinant in the outcome of viral infection. PMID- 2986949 TI - Effects of epidermal growth factor, phorbol myristate acetate, and arachidonic acid on choriogonadotropin secretion by cultured human choriocarcinoma cells. AB - Epidermal growth factor (EGF) binds specifically (Ka = 4 X 10(9) M-1; 1.3 X 10(11) receptors/mg cellular protein) to JEG-3 cells, which respond in the succeeding 24 h by a 400% increase in hCG secretion without a significant change in cell number. Since JEG-3 cells store less than 2% of the 24-h hCG secretion, a significant increase in hCG in the culture medium reflects increased hCG biosynthesis. It is observed that a tumor promoter, phorbol myristate acetate (PMA), binds specifically to the cells (Ka = 5 X 10(8) M-1; 3.9 X 10(11) receptors/mg), reduces (33%) the affinity but not the number of EGF receptors, and stimulates hCG to the same extent as does EGF. The relative potencies of PMA (100%), phorbol dibutyrate (25%), and nonesterified phorbol (no effect) to stimulate hCG parallel their known tumor-promoting activities. Since phospholipids and fatty acids have been implicated in the mechanism of action of EGF and PMA, the effects of arachidonic acid (AA) and inhibitors of its metabolism were studied. The addition of AA had no effect on basal or PMA stimulated hCG secretion, but it potentiated the effect of EGF by 200%. Indomethacin (a cyclooxygenase inhibitor) had an effect similar to that of AA. Nordihydroguairetic acid (a cyclooxygenase and lipoxygenase inhibitor) reduced by 90% basal and EGF- or PMA-stimulated hCG secretion. These results indicate that the AA pathway can influence the effects of EGF and PMA on hCG secretion and suggest an intermediary role for the lipoxygenase system. PMID- 2986951 TI - Modulatory effect of thyroid hormones on uptake of phosphate and other solutes across luminal brush border membrane of kidney cortex. AB - The mechanism whereby thyroid hormones modulate the transport properties of luminal brush border membrane (BBM) of renal proximal tubules was studied in thyroparathyroidectomized rats. Administration of both T4 and T3 increased BBM capacity for Na+ gradient-dependent uptake of phosphate (Pi) by BBM vesicles (BBMV). This effect of thyroid hormones was present in thyroparathyroidectomized and hypophysectomized rats, and it was not blocked by a saturating dose of propranolol. The stimulatory effect of T3 and T4 on BBM transport of Pi was dose dependent in the range of 5.2-520 nmol/100 g BW. Pretreatment of rats with inhibitors of 5'-monodeiodinase (5'-DI), iopanoic acid or ipodate, prevented the increase in serum T3 in rats injected with T4, but it did not diminish the increase in BBM transport of Pi. Administration of iopanoic acid and ipodate also prevented a 5-fold increase in 5'-DI activity in renal cortical tissue elicited by T4 administration. Treatment with T3 resulted in an increase of Pi transport across BBM from kidneys of rats subjected to dietary Pi deprivation due either to total fasting or to feeding of a low Pi diet. Further, T3 administration enhanced amiloride-sensitive Na+-H+ countertransport across BBM, but the uptake of 22Na+ by BBMV in the absence of pH gradient was not changed. The Na+ gradient-dependent uptake of L-[3H]proline by BBMV was slightly decreased, but the uptake of [14C]citrate was not changed in response to T3. Administration of T3 increased Pi transport in BBMV prepared from juxtamedullary cortex, but not in BBMV from superficial cortex. Conversely, the rate of Na+-H+ countertransport was enhanced, and the enzymatic activity of alkaline phosphatase was decreased in BBMV from superficial cortex; no changes in these parameters were found in BBMV from juxtamedullary cortex. Our results indicate that the stimulatory effect of administered T3 and/or T4 on renal BBM transport of Pi is distinct from the modulatory effect of dietary Pi intake and is not secondary to the beta-effect of catecholamines or to altered secretion of GH. Both T3 and T4 elicit the increase in BBM transport of Pi in a similar manner; T4 is effective even after profound inhibition of in vivo conversion of T4 to T3 by potent 5'-DI inhibitors. Administration of T3 stimulates Na+ gradient-dependent Pi uptake only in BBMV prepared from the juxtamedullary zone, and it stimulates the H+-Na+ countertransport only in BBMV from the outer cortical zone. These findings indicate that thyroid hormones modulate the two distinct transport functions of BBM derived from two different populations of renal proximal tubules. PMID- 2986952 TI - Endocrinological control and cellular localization of rat testicular angiotensin converting enzyme (EC 3.4.15.1). AB - Hypophysectomy of prepubescent (3-week-old) rats prevented the pubertal development of testicular, but not pulmonary, angiotensin-converting enzyme (EC 3.4.15.1). Additionally, hypophysectomy resulted in a loss of testicular converting enzyme activity in 10-week-old rats that had achieved puberty and had developed enzyme activity. Hormone regimens consisting of FSH/LH (7.5 U/rat X day), hCG (10 U/rat X day), or testosterone (1 mg/rat X day) were employed to ascertain their ability to maintain activity in hypophysectomized rats. All three of the above hormone regimens, if initiated on the first day after hypophysectomy of 10-week-old rats, were capable of maintaining testicular converting enzyme activity. Centrifugal elutriation of dispersed testicular cells indicated that the majority of enzyme activity in mature rats was associated with the germinal cells, a result consistent with the data accumulated from the hormonal studies. Lastly, [3H]captopril bound specifically to cellular fractions enriched in germinal cells. The above studies suggest that the pituitary gland is required for the development and maintenance of testicular angiotensin-converting enzyme in the rat by stimulating steroidogenesis in the testes. Furthermore, the sensitivity of converting enzyme activity to androgen coupled with the centrifugal elutriation and [3H] captopril binding studies strongly support the notion that testicular converting enzyme is associated with germinal cells. PMID- 2986953 TI - Effects of synthetic analogs and fragments of bovine parathyroid hormone on adenosine 3',5'-monophosphate level, ornithine decarboxylase activity, and glycosaminoglycan synthesis in rabbit costal chondrocytes in culture: structure activity relations. AB - Previously, we demonstrated that PTH increases the level of cAMP, the activity of ornithine decarboxylase (ODC; EC 4.1.1.17; which is a rate-limiting enzyme in polyamine biosynthesis), and glycosaminoglycan (GAG) synthesis (which is characteristic of the chondrocyte phenotype) in rabbit costal chondrocytes in culture. These studies suggested that the accumulation of cAMP and the induction of ODC by PTH are good markers of the differentiated phenotype of cultured chondrocytes. In the present study, the biological effects of a series of bovine PTH (bPTH) fragments and analogs on these three parameters of PTH action were examined. bPTH-(1-34), [Nle8,Nle18,Tyr34]bPTH-(1-34) amide and bPTH-(1-27) amide increased cAMP levels, ODC activity, and GAG synthesis in a dose-dependent manner over the concentration range of 10(-9)-10(-5) M. The order of decreasing potency was: bPTH-(1-34), [Nle8,Nle18,Tyr34]bPTH-(1-34) amide, and bPTH-(1-27) amide. On the other hand, [Nle8,Nle18,Tyr34]bPTH-(3-34) amide, bPTH-(5-27) amide, and [Tyr34]bPTH-(20-34) amide failed to increase cAMP levels, ODC activity, or GAG synthesis when present in concentrations up to 10(-5) M. However, [Nle8,Nle18,Tyr34]bPTH-(3-34) amide, bPTH-(5-27) amide, and [Tyr34]bPTH-(20-34) amide inhibited bPTH-(1-34)-stimulated increases in cAMP and ODC activity. These results partially define the principal structural determinants within the PTH molecule required for biological activity and expression of the differentiated phenotype of chondrocytes. PMID- 2986954 TI - Progesterone withdrawal stimulates mammary gland tubulin polymerization in pregnant rats. AB - The size of the polymerized tubulin pool in mammary glands of late pregnant rats increased 2.4-fold 24 h after bilateral ovariectomy using a [3H]colchicine binding assay for tubulin. The total (free plus polymerized) tubulin concentration was not altered. Stimulation of tubulin polymerization as well as the induced rise in lactose content were both completely inhibited in rats that had been fitted with progesterone implants at the time of surgery. The increase in polymerized tubulin was not blocked by 17 beta-estradiol implants, although this treatment did partially inhibit the rise in tissue lactose following ovariectomy. The results indicate that increased polymerization during induced lactogenesis reflects a shift of free tubulin into the polymerized pool and that this shift is related to the withdrawal of progesterone. PMID- 2986955 TI - Characterization and subcellular distribution of somatogenic receptor in rat liver. AB - Binding of [125I]iodobovine GH [( 125I]iodo-bGH) to rat liver microsomes and Golgi/endosomal fractions isolated from male and female rats has been characterized. Binding of bGH to a pure somatogenic site was suggested by the finding that 50% inhibition of [125I]iodo-bGH binding required 5-130 ng bGH, rGH, or hGH/incubation, while around 500 ng rat PRL/incubation were needed to obtain the same effect. Binding of [125I]iodo-bGH to microsomes and Golgi/endosomes was time, temperature, and protein dependent. Maximal specific binding occurred at 15 16 and 15-20 h at 22 C in Golgi and microsomal membranes, respectively. Subcellular distribution studies demonstrated in the Golgi/endosomal fractions compared to the total particulate fraction, while residual microsomes devoid of Golgi/endosomal-derived components were approximately 2-fold enriched. Low levels of somatogenic receptors were detected in lysosome-enriched fractions. Removal of endogenous ligand by treating Golgi/endosomal membranes with 3 M MgCl2 increased specific binding of bGH about 2- to 3-fold. These results indicate that approximately 50% of specific somatogenic binding sites in the low density fractions represent internalized ligand-receptor complexes. The level of rat liver somatogenic receptors did not show a pronounced sex differentiation; however, an endocrine dependence of somatogenic receptor levels is suggested by the finding that livers from rats in the late stages of pregnancy had a level of somatogenic receptors exceeding that of nonpregnant rats. PMID- 2986957 TI - Characterization of the 3,3',5-triiodo-L-thyronine-binding site on plasma membranes from human placenta. AB - The binding of [125I]T3 to sites on human placenta plasma membranes was characterized, and the binding site was solubilized after affinity labeling with N-bromoacetyl-[125I]T3 (BrAc[125I]T3). Two classes of T3-binding sites were detected. One class has a high affinity (Kd = 2.0nM) and a low capacity (approximately 320 fmol/mg protein); the other has a low affinity (Kd = 18.5 microM) and a high capacity (approximately 2.2 pmol/mg protein). The binding sites were found to be specific for T3 in that other thyroid hormone analogs (D T3, rT3, D-T4, and L-T4) were less effective or ineffective in displacing the bound [125I]T3. The affinity labeling ligand BrAc[125I]T3 was found to specifically label a protein with an apparent mol wt of 65,000, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The BrAc[125I]T3-labeled protein was solubilized with 2 mM 3-[( 3 cholamidopropyl)dimethylammonio]1-propane sulfonate. The apparent mol wt of the labeled protein was between 140,000 and 150,000 by Sephadex-G-200 gel filtration. These data demonstrate that a high affinity binding site specific for T3 is present on plasma membranes from human placenta and that the binding site is a protein, most likely a dimer, with a native mol wt between 140,000 and 150,000. PMID- 2986956 TI - Prolactin-binding components in rabbit mammary gland: characterization by partial purification and affinity labeling. AB - The molecular characteristics of the PRL receptor isolated from rabbit mammary gland microsomes were investigated. Two approaches were employed: 1) affinity purification of PRL receptors and direct electrophoretic analysis, and 2) affinity cross-linking of microsomal receptors with [125I]ovine PRL [( 125I]oPRL). PRL receptors were solubilized from mammary microsomes with 3-[( 3 cholamidopropyl)dimethylammonio]1-propane sulfonate and purified using an oPRL agarose affinity column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of the gel revealed at least nine bands, including a 32,000 mol wt band which was most intensively labeled with 125I using the chloramine-T method. Covalent labeling of PRL receptors with [125I]oPRL was performed using N-hydroxysuccinimidyl-4-azido benzoate, disuccinimidyl suberate, or ethylene glycol bis (succinimidyl succinate). A single band of 59,000 mol wt was produced by all three cross-linkers when sodium dodecylsulfate-polyacrylamide gel electrophoresis was performed under reducing conditions. Assuming 1:1 binding of hormone and binding subunit and by subtracting the mol wt of [125I]oPRL, which was estimated from the migration distance on the gel, the mol wt of the binding subunit was calculated as 32,000. In the absence of dithiothreitol during electrophoresis, only one major hormone-receptor complex band was observed. The same mol wt binding components were also detected in microsomal fractions of rabbit kidney, ovary, and adrenal. A slightly higher mol wt binding subunit was observed in rat liver microsomes. Rabbit liver microsomes revealed five [125I]oPRL-binding components, three of which were considered to be those of a GH receptor. Moreover, affinity labeling of detergent-solubilized and affinity purified mammary PRL receptors showed a similar major binding subunit. From these observations, we conclude that this predominant 32,000 mol wt component is a major binding subunit of the PRL receptor molecule and does not aggregate with itself or with other subunits through S-S linkages. PMID- 2986958 TI - Studies on the effects of catecholestrogens on prolactin secretion by cultured normal rat anterior pituitary cells. AB - The catecholestrogens 2-hydroxyesterone (2-OHE1) and 2-hydroxyestradiol (2-OHE2) are shown to directly inhibit PRL release by cultured normal rat anterior pituitary cells if they have been cultured in charcoal-treated estrogen-stripped fetal calf serum. Both catecholestrogens affect PRL release in a dose-dependent bimodal way. 2-OHE1 (1 microM) maximally inhibits PRL release by 38%, but 10 microM of the substance has no effect. In contrast, 0.1 microM 2-OHE2 maximally inhibits PRL release by 37%, while the effect of 1 microM of the compound was significantly less (-26%). The effect of the catecholestrogens on PRL release by cultured pituitary cells was significant after 2 h and was still effective after 48 h. The mechanism of action of 2-OHE1 and 2-OHE2 probably does not involve dopamine receptors, as 50 nM haloperidol did not alter the PRL release inhibitory effect of these substances. Moreover, 2-OHE1 and 2-OHE2 did not affect dopamine mediated inhibition of PRL release. The total amount of PRL (medium plus cells) present in cultures of normal pituitary cells exposed to 2-OHE1 or 2-OHE2 was identical to that in control cells. TRH (100 nM)-stimulated PRL release from cells exposed to catecholestrogens was significantly higher than that from control cells. Addition of the calcium entry-blocking agent verapamil prevented the inhibitory effect of dopamine on PRL release, but did not prevent the effects of 2-OHE1 and 2-OHE2. Preincubation of cultured normal rat pituitary cells for 30 min with 50 nM estradiol completely prevented the inhibitory effects of 2-OHE1 and 2-OHE2 on PRL release. In conclusion, the acute inhibitory effects of the catecholestrogens 2-OHE1 and 2-OHE2 on PRL release by cultured normal rat pituitary cells do not involve dopamine receptors or calcium transport over the cell membrane. These effects are only demonstrable after culture of the cells in charcoal-treated fetal calf serum and can be completely prevented by short term preincubation with estradiol. Our observations support the suggestion that the action of estradiol on lactotropic cells may be modulated by its metabolites, which have opposite effects on PRL release. PMID- 2986959 TI - Calcitonin gene-related peptide mimics calcitonin actions in brain on growth hormone release and feeding. AB - We have examined the capacity of calcitonin gene-related peptide (CGRP) to exert two biological actions in the central nervous system (CNS): modulation of GH release and alteration of food intake. These effects were compared with those of calcitonin (CT). Intracerebroventricular administration of both rat (r) CGRP (10 micrograms/10 microliter) and salmon (s) CT (250 ng/10 microliter) to chronically cannulated freely-moving rats reduced the frequency and amplitude of spontaneous GH secretory pulses; however, the suppressive effect was of longer duration with sCT than with rCGRP. Both peptides also significantly reduced spontaneous food intake over a 6-h period when administered centrally, although the decrease with sCT was again more extensive than that with rCGRP. The results demonstrate that CGRP can simulate two of the effects of CT in brain. Furthermore, the findings suggest that CGRP may function as an endogenous ligand for CT receptors in the CNS, and participate centrally in the modulation of GH release and the control of satiety. PMID- 2986960 TI - Central kappa- and mu-opiate receptors mediate ACTH-release in rats. AB - The control of ACTH secretion by opiates seems to involve stimulatory and inhibitory pathways, since opiate agonists and antagonists are capable of releasing ACTH in conscious rats. To elucidate the role of different opiate receptors in the control of ACTH release, rats were treated with receptor selective opiate agonists and antagonists. The mu-opiate agonists, morphine and (D-Ala2, MePhe4, Gly5-ol)enkephalin, and the benzomorphan kappa-opiate agonists, MR 2034 and MRZ 2549, both stimulated ACTH release after central or peripheral injection. The effects of morphine, but not those of MR 2034, were blocked by a low dose of naloxone (50 micrograms/kg) and by the mu-receptor antagonist, beta funaltrexamine. A 20 times higher dose of naloxone also blocked the effects of the kappa-agonist. Our data suggest that both mu- and kappa-opiate receptors are involved in the stimulation of ACTH release in rats. PMID- 2986961 TI - Isolation and characterization of a rat lung fraction enriched in alveolar wall basement membranes. AB - Separation of lung alveolar basement membranes from interstitial connective tissue protein has proved difficult, and a pure preparation of alveolar wall basement membranes (AWBM) is not available. We have modified a technique employing the detergent Triton X-100 for isolating AWBM from rat lungs by adding a step utilizing human skin collagenase (HSC), a highly purified enzyme obtained from skin fibroblasts that specifically cleaves non-basement membrane collagens. Triton extraction of both lungs yields 15-20 mg of basement membrane-enriched material referred to as crude fraction (CF). Ultrastructural studies show that CF includes both epithelial and endothelial basement membranes that appear similar to their in vivo counterparts and contain heparan sulfate proteoglycans. Extraction of type IV collagen is documented by the appearance of highly glycosylated hydroxylysine. This CF contains minimal amounts of contaminating elastin but significant amounts of interstitial collagens. CF was further purified for biochemical studies by incubation with HSC. HSC solubilized 20% of CF hydroxyproline resulting in a final fraction highly enriched in AWBM. Lung minces incubated in tritiated lysine produced a CF extract rich in newly formed type IV collagen, showing that lung tissue synthesizes AWBM collagen in vitro. PMID- 2986962 TI - Distinction and similarity in the structure of histones H1 and H5 as indicated by 13C nuclear-magnetic-resonance spectroscopy. AB - The 13C nuclear magnetic resonance studies have been carried out on histones H1 and H5, by focusing our interest on possible formation of specific salt bridges between acidic and basic amino acid residues in the proteins and also on the structural difference between the two proteins. The 13C chemical shift and pKa values of the carboxyl group of glutamic acid residues in the histones coincided with those of free glutamic acid. Based on this result and another experiment using completely modified lysine residues in the histones, no evidence for a specific interaction between acidic and basic residues has been found. It has also been shown that the pH-effects of aliphatic and aromatic resonances are quite different between H1 and H5, suggesting that the globular domain of H5 is more stable than that of H1. The correlation time (1.5 ns) for the alpha-carbons of H5 estimated from 13C nuclear Overhauser enhancement was twice as long as that of H1 (0.9 ns), indicating that the backbone in the N-terminal and C-terminal domains of H5 is less mobile than that of H1. PMID- 2986963 TI - Structural studies on the acidic polysaccharide of Bacillus cereus AHU 1356 cell walls. AB - Structural studies were carried out on the acidic polymer fraction isolated from lysozyme digests of the N-acetylated cell walls of Bacillus cereus AHU 1356. The acidic polymer fraction contained glucosamine, galactose, rhamnose, glycerol and phosphorus in a molar ratio of 1:1:2:1:1, together with small amounts of glycopeptide components and muramic acid 6-phosphate. The hydrogen fluoride treatment led to removal of glycerol and phosphorus from the polymer without loss of other components. Results of the NaIO4 oxidation, methylation and proton magnetic resonance spectroscopy of the native and dephosphorylated preparations, in combination with data of the analysis of oligosaccharides obtained from partial hydrolysis of polysaccharide, led to the most likely structure of the repeating units of the acidic polysaccharide chain, ----4)N-acetylglucosaminyl (alpha 1----3)rhamnosyl(alpha 1----3)galactosyl(alpha 1----4)[sn-glycerol 1 phospho-2]rhamnosyl(alpha 1----. PMID- 2986964 TI - A microbore high-performance liquid chromatography strategy for the purification of polypeptides for gas-phase sequence analysis. Structural studies on the murine transferrin receptor. AB - We describe herein the use of reversed-phase high-performance liquid chromatography coupled with the novel application of short (10 cm or less) microbore columns (2 mm internal diameter) to fractionate and purify a number of tryptic peptides generated from approximately 200 pmol purified murine transferrin receptor. The use of reversed-phase microbore columns permits the recovery of submicrogram amounts of purified polypeptides in high yield (greater than 90%) in small eluent volumes (20-60 microliter). In this manner, purified polypeptides can be loaded directly onto the gas-phase sequencer without further manipulation. This procedure avoids sample loss, which frequently occurs with other forms of concentration (e.g. lyophilization, evaporation). The application of second-order-derivative ultraviolet spectroscopy, using a diode array detector, for the analysis of aromatic aminoacid-containing peptides in complex tryptic digests is described. N-terminal amino acid sequence analyses were performed on six tryptic peptides, yielding 105 unique assignments; this corresponds to approximately 14% of the molecule. A comparison of this amino acid sequence information with the primary structure of human transferrin receptor deduced from the mRNA sequence [Nature (Lond.) 311, 675-678 (1984); Cell 39, 267 274 (1984)] reveals, with the exception of one tryptic peptide, a very close sequence homology between the murine and human transferrin receptors. PMID- 2986965 TI - ATP synthesis in Methanobacterium thermoautotrophicum coupled to CH4 formation from H2 and CO2 in the apparent absence of an electrochemical proton potential across the cytoplasmic membrane. AB - Methanogenic bacteria are considered to couple methane formation with the synthesis of ATP by a chemiosmotic mechanism. This hypothesis was tested with Methanobacterium thermoautotrophicum. Methane formation from H2 and CO2 (2.5 - 3 mumol X min-1 X mg cells-1) by cell suspensions of this organism resulted in the formation of an electrochemical proton potential (delta mu H +) across the cytoplasmic membrane of 230 mV (inside negative) and in the synthesis of ATP up to an intracellular concentration of 5 - 7 nmol/mg. The addition of ionophores at concentrations which completely dissipated delta mu H + without inhibiting methane formation did not result in an inhibition of ATP synthesis. It thus appears that delta mu H + across the cytoplasmic membrane is not the driving force for the synthesis of ATP in M. thermoautotrophicum. PMID- 2986967 TI - The role of the Na+/H+ exchange system in the regulation of the internal pH in cultured cardiac cells. AB - The Na+/H+ exchange system is not the major mechanism that regulates the internal pH value (pHi) of chick cardiac cells in culture under normal physiological conditions in the absence of carbonate. In cardiac cells in which the internal pH has been lowered to 6.6-6.7, the Na+/H+ exchanger becomes the major mechanism to bring back pHi to normal values (pHi = 7.3). The blockade of the Na+/H+ exchange activity with an active amiloride derivative, ethylisopropylamiloride, prevents internal pH recovery. The internal pH dependence of the Na+/H+ exchanger activity has been carefully studied. The [H+]i-dependence is very cooperative. For an external pH of 7.4, the system is nearly completely inactive at pHi 7.8 and nearly completely active at pHi 6.9-7.0 with half-maximum activation at pHi = 7.35. The increased activity of the Na+/H+ exchange system which follows the acidification of the internal medium produces an activation of the (Na+,K+) ATPase. PMID- 2986966 TI - Effect of cGMP and cations on the permeability of cattle retinal disks. AB - Guanosine 3',5'-monophosphate and Na or Ca ions affect the transmembrane movements of the same pool of intradiskal ions. Extradiskal Na ions activate the efflux of intradiskal Na ions. Extradiskal Ca ions activate the efflux of intradiskal Rb ions. Na and Ca ions activate Na/Ca or Ca/Ca exchange, as previously described. cGMP activates a membrane permeability for all the cations tested, as previously described. The reciprocal relations between cGMP and the other pathways for ion movements through disk membranes are systematically examined. Some analogies between the cGMP-activated permeability of the disk membranes and the light-sensitive conductance of the rod plasma outer membrane are discussed. PMID- 2986968 TI - Catabolism of Ap3A and Ap4A in human plasma. Purification and characterization of a glycoprotein complex with 5'-nucleotide phosphodiesterase activity. AB - A hydrolase splitting adenosine(5')triphospho(5')adenosine (Ap3A) to AMP and ADP has recently been detected in human plasma [Luthje, J. and Ogilvie, A. (1984) Biochem. Biophys. Res. Commun. 118, 704-709]. The enzyme has been purified to apparent homogeneity, as stained in a native polyacrylamide gel. From gel filtration data a Stokes radius of 5.9 nm was calculated, suggesting a molecular mass of about 230 kDa. The presence of the non-ionic detergent Triton X-100 did not change the molecular mass. The hydrolase dissociated to three major protein components (66 kDa; 45 kDa; 16 kDa) during polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and mercaptoethanol. Binding of the native enzyme to concanavalin-A--Sepharose and specific inhibition of binding by methyl mannoside indicated that the hydrolase is a glycoprotein. Two of the subunits (66 kDa; 45 kDa) could be affinity-labeled with radioiodinated concanavalin A. Active hydrolase could be prepared in buffers without added metal ions. Treatment with EDTA, however, completely abolished the hydrolytic activity. The enzyme could be reactivated by incubation with Ca2+, Co2+ and, at best, with Zn2+, whereas Mg2+ was ineffective. The affinity of the enzyme for Ap3A was high (Km = 1 microM), with normal Michaelis-Menten kinetics. The homolog dinucleotide Ap4A was also substrate (Km = 0.6 microM) yielding AMP and ATP as products after the asymmetric split. Other dinucleotides, such as NAD, and also mononucleotides (ATP,UTP) were degraded to nucleoside monophosphates indicating a broad specificity of the enzyme. The synthetic compound thymidine 5'-monophosphate p-nitrophenyl ester was substrate with low affinity whereas its 3'-homolog was not hydrolyzed. Optimal activity of the hydrolase was found at pH 8.5. PMID- 2986969 TI - Isolation and properties of cytochrome c oxidase from rat liver and quantification of immunological differences between isozymes from various rat tissues with subunit-specific antisera. AB - Cytochrome c oxidase was isolated from rat liver either by affinity chromatography on cytochrome-c--Sepharose 4B or by chromatography on DEAE Sepharose. Dodecyl sulfate gel electrophoresis of both preparations showed the same subunit pattern consisting of 13 different polypeptides. Kinetic analysis of the two preparations gave a higher Vmax for the enzyme isolated by chromatography on DEAE-Sephacel. Specific antisera were raised in rabbits against nine of the ten nuclear endoded subunits. A monospecific reaction of each antiserum with its corresponding subunit was obtained by Western blot analysis, thus excluding artificial bands in the gel electrophoretic pattern of the isolated enzyme due to proteolysis, aggregation or conformational modification of subunits. With an antiserum against rat liver holocytochrome c oxidase a different reactivity was found by Western blot analysis for subunits VIa and VIII between isolated cytochrome c oxidases from pig liver or kidney and heart or skeletal muscle. For a quantitative analysis of immunological differences a nitrocellulose enzyme linked immunosorbent assay was developed. Monospecific antisera against 12 of the 13 subunits of rat liver cytochrome c oxidase were titrated with increasing amounts of total mitochondrial proteins from different rat tissues dissolved in dodecyl sulfate and dotted on nitrocellulose. The absorbance of a soluble dye developed by the second peroxidase-conjugated antibody was measured. From the data the following conclusions were obtained: (a) The mitochondrial encoded catalytic subunits I-III of cytochrome c oxidase are probably identical in all rat tissues. (b) All nine investigated nuclear encoded subunits of cytochrome c oxidase showed immunological differences between two or more tissues. Large immunological differences were found between liver, kidney or brain and heart or skeletal muscle. Minor but significant differences were observed for some subunits between heart and skeletal muscle and between liver, kidney and brain. (c) Between corresponding nuclear encoded subunits of cytochrome c oxidase from fetal and adult tissues of liver, heart and skeletal muscle apparent immunological differences were observed. The data could explain cases of fatal infantile myopathy due to cytochrome c oxidase deficiency. PMID- 2986970 TI - Fractionation by high-performance liquid chromatography of the low-molecular-mass high-mobility-group (HMG) chromosomal proteins present in proliferating rat cells and an investigation of the HMG proteins present in virus transformed cells. AB - The low-molecular-mass high-mobility-group (HMG) proteins from young rat thymus nuclei were fractionated by high-performance liquid chromatography. Two proteins analogous to calf HMG14 and HMG17 were found together with a third major component HMGI similar to that found in HeLa cells [Lund et al. (1983) FEBS Lett. 152, 163-167]. HMGI has as amino acid composition similar to but distinct from HMG14 and HMG17. The three proteins form a family of proteins with HMG14 having an amino acid composition intermediate between HMG17 and HMGI. HMGI is present in proliferating fibroblasts and embryos but is present in very low levels in rat liver, a non-dividing tissue, supporting the notion that HMGI is required for proliferating cells. Fibroblasts transformed with avian sarcoma virus have high levels of HMGI and an additional band HMGI' but the presence of HMGI and HMGI' is not dependent on a functional src gene product. PMID- 2986971 TI - Spin-label study of conformational changes induced by pH and ligands in leghemoglobin from yellow lupine root nodules. AB - Conformational changes induced by ligands and pH in lupine ferrileghemoglobin selectively modified at Tyr-E16 by the imidazolide spin label has been studied by the method of electron spin resonance in the pH range 6-13. It is shown that in the alkaline pH region the bound spin label registers a local conformational transition which precedes the alkaline denaturation of the protein. In aquamet, cyanide and nicotinate complexes of ferrileghemoglobin this transition occurs with pK 10.5, in acetate and azide complexes with pK 11. In all these ligand derivatives the transition is induced by alteration in the ionization state of one group (delta nH+ approximately equal to 1), most probably, the epsilon-amino group of Lys-GH3. The latter is linked with the Glu-A14 residue and this bond is essential for maintaining the native conformation of leghemoglobin. The ligand induced conformational changes in the vicinity of the label are small and consist, most likely, in some alteration of the mutual arrangement of the AE and GH helical complexes. No correlation has been revealed between the spin state of the heme iron and the conformation of leghemoglobin in the region under study. PMID- 2986972 TI - The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora, determined by cDNA and gene sequencing. AB - The primary structure of the iron-sulfur subunit of ubiquinol-cytochrome c reductase from Neurospora mitochondria was determined by cDNA and genomic DNA sequencing. A first cDNA was identified from a cDNA bank cloned in Escherichia coli by hybridization selection of mRNA, cell-free protein synthesis and immunoadsorption. Further cDNA and geonomic DNA were identified by colony filter hybridization. The N-terminal sequence of the mature protein was determined by automated Edman degradation. From the sequence a molecular mass of 24749 Da results for the precursor protein and of 21556 Da for the mature protein. The presequence consists of 32 amino acids with four arginines as the only charged residues. The mature protein consists of 199 amino acids. It is characterized by a small N-terminal hydrophilic part of 29 residues, a hydrophobic stretch of 25 residues and a large C-terminal hydrophilic domain of 145 residues. The only four cysteines of the protein, which are assumed to bind the 2 Fe-2S cluster, are located in a moderate hydrophobic region of this large domain. Cysteines 3 and 4 are unusually arranged in that they are separated by only one proline. From sequence data the arrangement of the subunit in the membrane is deduced. PMID- 2986974 TI - The protein phosphatases involved in cellular regulation. Influence of polyamines on the activities of protein phosphatase-1 and protein phosphatase-2A. AB - The effects of polyamines on the oligomeric forms of protein phosphatase-1 (1G), protein phosphatase-2A (2A0, 2A1 and 2A2) and their free catalytic subunits (1C and 2AC) has been studied using homogeneous enzymes isolated from rabbit skeletal muscle. Spermine increased the activity of protein phosphatase-2A towards eight of nine substrates tested. Half-maximal activation was observed at 0.2 mM with optimal effects at 1-2 mM. Above 2 mM, spermine became inhibitory. The most impressive activation of protein phosphatase-2A was obtained with glycogen synthase, especially when phosphorylated at sites-3 (8-15-fold with protein phosphatase-2A1) and phenylalanine hydroxylase (6-7-fold with protein phosphatase 2A1) as substrates. Activation of protein phosphatases 2A0, 2A1 and 2A2 was greater than that observed with 2AC. Spermine was a more potent activator than spermidine, while putrescine had only a small effect. Qualitatively similar results were obtained with five other substrates, although maximal activation was much less (1.3-3-fold with protein phosphatase-2A1). The rate of dephosphorylation of glycogen phosphorylase was decreased by spermine, inhibition being more pronounced with protein phosphatase-2AC than with 2A0, 2A1 and 2A2. Spermine (I50 = 0.1 mM with protein phosphatase-2AC) was a more potent inhibitor than spermidine (I50 = 0.9 mM) or putrescine (I50 = 8 mM). Partially purified preparations of protein phosphatases-2A0, 2A1 and 2A2 from from rat liver were affected by spermine in a similar manner to the homogeneous enzymes from rabbit skeletal muscle. Spermine did not activate protein phosphatase-1 to the same extent as protein phosphatase-2A. Greatest stimulation (2.5-fold) was again observed with glycogen synthase labelled in sites-3, with half-maximal activation at 0.2 mM and optimal effects at 1-2 mM spermine. Spermine was a much more effective stimulator than spermidine, while putrescine was ineffective. Very similar results were obtained with protein phosphatases 1G and 1C. With four other substrates maximal activation by spermine was less than 1.5-fold, while the dephosphorylation of glycogen synthase (labelled in site-2), phosphorylase kinase, pyruvate kinase and glycogen phosphorylase were inhibited. Spermine (I50 = 0.04 mM) was a more potent inhibitor of the dephosphorylation of glycogen phosphorylase than spermidine (I50 = 0.9 mM) or putrescine (I50 = 9 mM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2986973 TI - The protein phosphatases involved in cellular regulation. Purification and characterisation of the glycogen-bound form of protein phosphatase-1 from rabbit skeletal muscle. AB - A type-1 protein phosphatase (protein phosphatase-1G) was purified to homogeneity from the glycogen-protein particle of rabbit skeletal muscle. Approximately 3 mg of enzyme were isolated within 4 days from 5000 g of muscle. Protein phosphatase 1G had a molecular mass of 137 kDa and was composed of two subunits G (103 kDa) and C (37 kDa) in a 1:1 molar ratio. The subunits could be dissociated by incubation in the presence of 2 M NaCl, separated by gel-filtration on Sephadex G 100, and recombined at low ionic strength. The C component was the catalytic subunit, and was identical to the 37-kDa type-1 protein phosphatase catalytic subunit (protein phosphatase-1C) isolated from ethanol-treated muscle extracts, as judged by peptide mapping. The G component was the glycogen-binding subunit. It was very asymmetric, extremely sensitive to proteolytic degradation, and failed to silver stain on SDS/polyacrylamide gels. Protein phosphatase-1G was inhibited by inhibitor-1 and inhibitor-2, but unlike protein phosphatase-1C, the rate of inactivation was critically dependent on the ionic strength, temperature and time of preincubation with the inhibitor protein. At near physiological temperature and ionic strength, protein phosphatase-1G was inactivated very rapidly by inhibitor-1. Protein phosphatase-1G interacted with inhibitor-2 (I-2) to form an inactive species, with the structure GCI-2. This form could be activated by preincubation with Mg-ATP and glycogen synthase kinase-3. The G subunit could be phosphorylated on a serine residue(s) by cyclic-AMP-dependent protein kinase, but not by phosphorylase kinase or glycogen synthase kinase-3. Phosphorylation was rapid and stoichiometric, and increased the rate of inactivation of protein phosphatase-1G by inhibitor-1. The relationship of the G subunit to the 'deinhibitor protein' is discussed. PMID- 2986975 TI - Catalytic properties of a human cytomegalovirus-induced protein kinase. AB - Human cytomegalovirus, a DNA virus whose genome contains a fragment of transforming DNA, induces a threonine-serine protein kinase having a molecular mass of 68 kDa (p68). p68 was extracted from cells 96-144 h after infection, and immunoprecipitated with a monoclonal antibody (F6b). Antibody-enzyme complexes were immobilized on heat/formaldehyde-inactivated Staphylococcus aureus. The best substrates for p68 were acidic proteins, phosvitin and casein. Glycogen synthase, phosphorylase alpha and histones were phosphorylated at rates not higher than 1 4% that obtained with phosvitin as substrate. ATP and GTP were equally good substrates of p68. p68 is able to autophosphorylate at the same residues (i.e. threonine and serine) as the protein substrates. Autophosphorylation does not seem to represent an intermediate in substrate phosphorylation. The protein kinase activity of p68 was not enhanced by cAMP, calcium ions, or polyamines like spermine or spermidine. Only at low Mg2+ concentration spermine enhanced by 68% the rate of casein phosphorylation. Heparin, a potent inhibitor of casein kinase II, inhibits p68 activity too, but ten-times higher concentrations were required for the same degree of inhibition. Quercetin, a bioflavonoid, acts as a strong inhibitor of p68 protein kinase activity. The inhibitory effect of quercetin was competitive towards the nucleotide substrate (Ki = 2.8 microM), and non competitive towards the protein substrate (Ki = 15 microM). PMID- 2986976 TI - Molecular forms of sphingomyelinase and non-specific phosphodiesterases in Epstein-Barr virus-transformed lymphoid cell lines from Niemann-Pick disease types A and B. AB - Molecular forms of sphingomyelinase and phosphodiesterases from lymphocytes- and Epstein-Barr virus-transformed lymphoid cell lines were separated by preparative electrofocusing in granulated gels. In either type of cell derived from normal individuals, sphingomyelinase focused as a single peak (pI = 5.60 +/- 0.1) while phosphodiesterases hydrolyzing bis(4-methylumbelliferyl)phosphate and bis(4 methylumbelliferyl)diphosphate separated into seven and three molecular forms respectively; one of the latter showed sphingomyelinase as well as phosphodiesterase activities. Lymphoid cell lines derived from patients with Niemann-Pick disease, types A or B, were practically devoid of sphingomyelinase activity; this was not so for the phosphodiesterases which focussed essentially as normal. The protein peak, which in normal cells contained the three activities, had phosphodiesterase but no sphingomyelinase activity in the Niemann Pick cells. In normal cells, sphingomyelinase and phosphodiesterase activities of this peak showed different responses to heating and several effectors. These data suggest that in lymphoid cell lines, which are a useful model for studies of Niemann-Pick disease, sphingomyelinase and phosphodiesterases are subject to separate genetic coding and that the latter activities are not a reliable measure for diagnosing Niemann-Pick disease. PMID- 2986977 TI - Computed tomography for staging bronchogenic carcinoma. AB - One hundred and fourty patients with a primary lung neoplasm underwent transmission computed tomography (TCT) for staging prior to therapy. TCT results were compared with the post-operative T- and N-stage. The preoperative TCT examination correctly classified 70% of all tumors. 73% of T1, 63% of T2 and 74% of T3 tumors did not require restaging after surgery. TCT correctly staged 80% of the patients with respect to the N-stage. Ninety-seven percent of the patients with N0, 42% with N1 and 79% with N2 tumors were correctly classified. Serious over- and understaging was observed in patients with centrally located bronchogenic carcinoma. This was due to the difficulty of distinguishing tumor tissue from enlarged lymph nodes and atelectasis. PMID- 2986978 TI - Importance and value of a preoperative diagnosis in oat cell carcinoma by radiography and its verification by fine needle biopsy (FNB). AB - Surgical resection currently offers the best hope of cure in Non Small Cell Lung Cancer (NSCLC), while chemotherapy and/or radiotherapy is the primary choice for Small Cell Lung Cancer (SCLC). The preoperative diagnosis of small cell anaplastic carcinoma (oat cell carcinoma) therefore is extremely important for adequate patient management treatment and prognosis. The patients history, clinical findings and radiographic patterns are all important for the differential diagnosis of SCLC (oat cell cancer) constituting a clinical, radiological and histopathological entity. FNB is a minor, fast, safe, inexpensive procedure, which is easy to perform, not uncomfortable for the patient and which, due to high accuracy may confirm or exclude SCLC (oat cell cancer). Major surgery with its inherent risk of morbidity/mortality may such be avoided allowing to choose the optimal and most appropriate form of alternative treatment permitting a high quality of life during the course of the disease. In a series of 2726 patients with 5300 FNB, of which 1264 (46%) showed evidence of malignancy SCLC (oat cell carcinoma) was cytologically diagnosed in 54 cases (6%). No serious complication occurred. PMID- 2986979 TI - Visualization of epidermal growth factor receptor in cryosections of cultured A431 cells by immuno-gold labeling. AB - Cryo-ultramicrotomy in combination with immuno-gold labeling has been demonstrated to present a powerful tool in the visualization of extra- and intracellular located antigens. We have applied this method to localize epidermal growth factor (EGF) receptor in cultured A431 human epidermoid carcinoma cells. However, both the labeling efficiency, maintenance of antigenicity, and the recognizability of the ultrastructure in cryosections are highly dependent upon the fixation procedures. Using 125I-EGF or a consecutive labeling with a monoclonal anti EGF-receptor antibody, rabbit-anti-mouse antibody and 125I protein A, it was shown that maintenance of antigenicity was optimal using 2% paraformaldehyde as a fixative, whereas under these conditions also the recognizability of ultrastructure was sufficient. After appropriate fixation and labeling, gold particles were observed associated with various regions of the plasma membrane, including coated pits, and with various types of vesicles, including coated vesicles, intracellular vesicular membranes, multi-vesicular bodies and lysosomes. The results indicate that this method allows a visualization of EGF-receptors and resolution of the EGF-receptor processing pathway at the electron microscopic level, independent of the internalization process of labeled ligands. PMID- 2986980 TI - Morphological analysis of ligand uptake and processing: the role of multivesicular endosomes and CURL in receptor-ligand processing. AB - The receptor-mediated endocytosis and intracellular processing of transferrin and mannose receptor ligands were investigated in bone marrow-derived macrophages, fibroblasts and reticulocytes. Mannosylated bovine serum albumin (BSA) conjugated to colloidal gold (Au-man-BSA) or colloidal gold-transferrin (AuTf) were used to trace ligand processing in these cells. These ligands appeared to be processed by mechanisms similar to those observed previously with other mannose receptor and galactose receptor ligand probes. After uptake via coated pits and coated vesicles, Au-man-BSA appeared in small uncoated vesicles and tubular structures and was transferred to large, sometimes multivesicular endosomes (MVEs), which sometimes had arm-like protrusions reminiscent of CURL (compartment of uncoupling of receptor and ligand) [10, 11]. Initially these structures became increasingly multivesicular, but during longer incubations the inclusion vesicles appeared to disintegrate to leave a denser, amorphous lumen. Inclusion vesicle disintegration may result from the introduction of lysosomal enzymes into these structures. These results suggest a model for differential receptor-ligand and ligand-ligand sorting. As suggested [10, 11] membrane constituents may be recycled to the plasma membrane from the arms of CURL. Receptor-bound ligands, such as transferrin, would also recycle. The luminal contents, including dissociated ligands, other soluble proteins and inclusion vesicles (containing some membrane proteins), would target to lysosomes. This would result in the lysosomal degradation of any membrane proteins that were incorporated in the inclusion vesicle membranes. PMID- 2986981 TI - Gap junctions between pancreatic B-cells are modulated by cyclic AMP. AB - Gap junction morphology was studied on freeze fracture replicas of pancreatic islet tissue, using morphometric techniques. In rat islets in situ, 60 percent of the connexions were polygonally packed in gap junctions, whereas the remaining part occurred in linear strands. After collagenase isolation, the islets presented similar numbers of gap junctions but contained virtually no linear strands. The distribution of connexions over polygonal or linear arrays also varied with the culture conditions: at 11.2 mM glucose, a higher percentage of particles occurred in gap junctions than at 5.6 mM glucose; this was also the case in other conditions with elevated cellular cyclic AMP levels. The total number of connexions increased when islets were cultured with dibutyryl cyclic AMP or with a phosphodiesterase inhibitor; conditions with an augmented number of gap junctions also displayed an elevated islet cyclic AMP content. A similar association was noted in newly formed aggregates of pancreatic B-cells purified by autofluorescence-activated. cell sorting. These results indicate that the number of classically defined gap junctions is not only dependent on the total number of connexions but also on their organization within the membrane. It is suggested that the distribution of connexions over polygonal and linear arrays follows a dynamic equilibrium varying with the extracellular conditions. Cyclic AMP appears to modulate the number of gap junctions between pancreatic B-cells both through an induction of new connexions and through an assembly of linearly organized particles into polygonal arrays. PMID- 2986982 TI - Acute ascending motor paralysis due to rabies: a clinicopathological report. AB - An unusual presentation of a not so uncommon disease is described. Practical difficulties in arriving at a diagnosis of paralytic rabies are discussed. An enquiry of exposure and suspicion of rabies in atypical Landry's paralysis is stressed. The importance of autopsy to confirm either of the diagnoses is emphasized. PMID- 2986983 TI - Immunofluorescence detection of liver cell membrane autoantibodies using PLC/PRF/5 cells. AB - Detection of liver cell membrane autoantibodies is routinely performed by immunofluorescence testing of patient sera on rabbit hepatocyte suspensions. We have investigated the possible use of cells from the PLC/PRF/5 human hepatoma cell-line. These cells were employed as substrate in an immunofluorescence test which was compared with the conventional rabbit hepatocyte assay. We found a close correlation between the results obtained with these different substrates on visual reading. We furthermore compared visual reading of immunofluorescence preparations with flow-cytometrical analysis of immunostained cell suspensions. The results with these different methods were largely confirmatory. The PLC/PRF/5 cells are easily available and should therefore be regarded as a highly valuable new substrate for detection of liver cell membrane autoantibodies. Flow cytometry appears to be a technically simple and reliable method for quantitative analysis. PMID- 2986985 TI - Abnormalities of in vitro responses to polyclonal activation of peripheral blood lymphocytes in patients with active sarcoidosis. AB - A variety of immunological abnormalities have been described in patients with sarcoidosis. At the blood level, both hypo and hyper immune responsiveness seem to coexist and were related to abnormal T cell and macrophage functions by using allogenic cocultures and/or lymphocyte fractionation. We tested several components of cell mediated responses with two in vitro models: (a) the pokeweed mitogen activation of B cells which is T cells and macrophage dependent; (b) the Epstein-Barr virus (EBV) activation of B cells which is T cell- and macrophage independent. We confirm previous data showing that PWM induced Ig Production of peripheral blood lymphocytes (PBL) of patients with sarcoidosis is significantly reduced compared with normal PBL. However, this is associated with an increase of IgG, IgM synthesis of EBV-infected PBL in these sixteen patients. Thus, there is no evidence for a complete B cell defect in sarcoid PBL. Furthermore, by using limiting dilution analysis of antibody secreting cells, there is an increase of precursor B cells EBV infectable in PBL of sarcoid patients but T cells are effective in reducing EBV-induced B cell proliferation. Finally, these abnormalities are concomitant to the disease, disappear with it, and are apparently not correlated with the stage or the activity of sarcoidosis. PMID- 2986984 TI - Abnormality of adenylate cyclase regulation in human platelet membranes in renal insufficiency. AB - Adenylate cyclase in human platelets is under dual control of prostaglandins (PGI2 and PGE1) and catecholamines. The adenylate cyclase complex in membranes of platelets from ten patients with uraemia was investigated. The activation of the platelet cyclase by PGE1 is increased in the uraemic state, Vmax 4436 +/- 607 pmol cAMP mg-1 15 min-1. In the normal state Vmax is 2098 +/- 309 pmol cAMP mg-1 15 min-1. The alpha 2-adrenergic receptor was assayed with 3H-yohimbine binding. The density of receptors was equal in the uraemic (175 fmol mg-1 membrane protein) and the normal (170 fmol mg-1 membrane protein) states. Norepinephrine/3H-yohimbine competition binding revealed that catecholamines were bound with normal affinity in platelets in uraemia. Yet the inhibition of adenylate cyclase through the alpha 2-adrenergic receptor was diminished since Vmax values of adenylate cyclase with PGE1 and PGE2 + norepinephrine did not significantly differ. In the normal state, norepinephrine significantly (P less than 0.05) inhibited the PGE1 stimulated cyclase. It is concluded that platelet adenylate cyclase in the uraemia has an increased capacity for activation which is the result of both a sensitized stimulatory mechanism (prostaglandin mediated) and a deficient inhibitory mechanism (catecholamine mediated). It is suggested that a defect exists in the inhibitory nucleotide binding protein (NI) which is the coupling unit between the adenylate cyclase catalytic subunit (C). PMID- 2986986 TI - Vitamin D3 uptake by the isolated perfused rat liver from lipoprotein fractions is separate from cholesterol and triglyceride uptake. AB - Using the isolated perfused rat liver we examined the uptake of [14C] or [3H] vitamin D3 and [14C] triglycerides or [3H] cholesterol by the liver of normal rats, from different lipoprotein fractions, as measured by disappearance from the perfusate. When incorporated into chylomicrons only 43% of the vitamin D3 remained in the perfusate at 60 min (i.e. 57% uptake) as compared to 68% of the triglycerides (i.e. 32% uptake). When added on very low density lipoproteins (VLDL) at 60 min 37 +/- 3% (n = 6) of the vitamin D3, 38 +/- 2% of the cholesterol (n = 3) (P NS), and 83 +/- 4% of the triglycerides (n = 3) remained in the perfusate (P less than 0.0005 for cholesterol:triglycerides and vitamin D3:triglycerides). For high density lipoprotein fraction (HDL) isolated perfused livers were studied with and without albumin present in the perfusate. Without albumin 19 +/- 8% (n = 3) of the vitamin D3 and 43 +/- 8% (n = 3) of the cholesterol remained in the perfusate at 60 min. The results with albumin present were 40 +/- 1% (n = 5) of the vitamin D3 and 63 +/- 4% (n = 5) of the cholesterol at 60 min (P less than 0.0005). The cholesterol:cholesterol ester ratio of the VLDL fraction was 8.8:1 and of the HDL fraction 1:1.4. There was no metabolism of vitamin D3 during the 1 h perfusion. These results suggest that vitamin D3 is in equilibrium with the lipoprotein surface, and the hepatic uptake of vitamin D is a surface phenomenon independent of lipoprotein metabolism. PMID- 2986987 TI - VLA-1: a T cell surface antigen which defines a novel late stage of human T cell activation. AB - The VLA-1 protein complex defines a previously undescribed very late stage of activated T cell differentiation, following either alloantigen or mitogen activation. This protein appears after 2-3 weeks of activation, considerably later than the early T cell activation antigens such as the interleukin 2 (IL 2) receptor, transferrin receptor, 4F2 antigen, T10 and HLA-DR, and has therefore been termed very late antigen-1 (VLA-1). Unlike the IL 2 receptor, VLA-1 expression does not require restimulation with antigen, and in fact, VLA-1 expression was high on T cells that had lost their IL 2 receptors. Expression of VLA-1 was found on all or nearly all long-term-activated T cells including T4+ and T8+ clones, bulk cultures, long-term T cells from adults and newborns and long-term T cells maintained in pure or crude IL 2 preparations. VLA-1 was also found on HTLV-1 infected T cell populations. Immunoprecipitation experiments confirmed that the VLA-1 protein complex (210 000/130 000 Mr) can be co-expressed with another protein complex called VLA-2 (165 000/130 000 Mr) on the same T cell clones. However, co-expression was not obligatory because in some long-term cultures little or no VLA-2 was present relative to VLA-1. Because VLA-1 defines a novel late stage of T cell activation, being present on all or most all types of long-term activated T cells, and not on any other cell types in peripheral blood, it has unique potential as a marker for activated T cells in vivo and may provide a clue towards elucidating novel long-term T cell functions or growth requirements of this late stage of T cell differentiation. PMID- 2986988 TI - Pharmacological and neurochemical properties of 1,4-diazepines with two annelated heterocycles ('hetrazepines'). AB - 1,4-Diazepines with two annelated heterocycles ('hetrazepines') such as brotizolam (WE 941), WE 973 and WE 1008 bind with high affinities to benzodiazepine receptors in the central nervous system. Brotizolam has a pharmacologic spectrum of action similar to clinically useful benzodiazepines, while the closely related derivatives WE 973 and WE 1008 appear to lack hypnotic action. Unlike other benzodiazepine receptor ligands which share common pharmacologic properties with the benzodiazepines, the apparent affinities of WE 973 and WE 1008 are not increased significantly in the presence of GABA, even at an elevated incubation temperature. Furthermore, the apparent affinities of these compounds do not appear to be reduced as a result of increasing the incubation temperature. Brotizolam, like the benzodiazepines, facilitates GABAergic transmission in zona recitulata neurons of the substantia nigra. In contrast, at a dose which inhibits cell firing, WE 973 does not appear to significantly augment the inhibitory action of GABA in these cells. These observations suggest that the so-called 'GABA shift' may not be a valid means of distinguishing benzodiazepine-like compounds in vitro. Furthermore, these data suggest that facilitation of GABAergic transmission may be necessary for the hypnotic action of benzodiazepine receptor ligands, but not for the anticonflict or the anticonvulsant actions of such compounds. PMID- 2986989 TI - (+)-[3H]SKF 10,047, (+)-[3H]ethylketocyclazocine, mu, kappa, delta and phencyclidine binding sites in guinea pig brain membranes. AB - Binding of the opiates (+)-[3H]SKF 10,047 [N-allylnormetazocine; (+)-[3H]SKF] and (+)-[3H]ethylketocyclazocine [(+)-[3H]EKC] were compared to mu, kappa and delta and phencyclidine (PCP) receptor binding in guinea pig brain membranes. (+) [3H]SKF and (+)-[3H]EKC binding were not blocked by naloxone, and had different drug selectivity compared to mu, kappa and delta binding sites. The number of binding sites, drug selectivity and region distribution in brain were similar for (+)-[3H]SKF and (+)-[3H]EKC. Sigma opiates that are associated with psychotomimetic activities, such as pentazocine, cyclazocine, SKF 10,047 and bremazocine, were potent inhibitors of (+)-[3H]SKF and (+)-[3H]EKC binding. Haloperidol was the most potent inhibitor of (+)-[3H]SKF binding. Haloperidol and sigma opiates demonstrated biphasic displacement of [3H]PCP binding, suggesting that [3H]PCP labelled two sites. PCP had a similar affinity for both (+)-[3H]SKF and [3H]PCP binding sites in the presence of 100 mM NaCl. The highest concentrations of (+)-[3H]SKF and (+)-[3H]EKC bindings sites were in the hypothalamus, anterior pituitary, midbrain, pons and medulla. PMID- 2986990 TI - Mu, but not kappa, opioid agonists induce contractions of the canine small intestine ex vivo. AB - The proposed kappa opioid receptor agonists ethylketocyclazocine (EK), nalorphine, bremazocine and U-50,488H were evaluated for their ability to produce contractions of isolated, vascularly perfused canine small intestinal segments. Responses to these agonists were compared to those of morphine and phenazocine, a mu benzomorphan. Morphine (0.04-25 micrograms) and phenazocine (0.01-3.0 micrograms) both produced naloxone-reversible contractions, suggesting that the responses were mediated largely by mu opioid receptors. In contrast, the proposed kappa agonists were ineffective in producing intestinal stimulation, with only EK (1-100 micrograms) showing minimal but significant activity at very high doses. We suggest that the effects of EK may be mediated through mu opioid receptors and that kappa receptors appear not to be involved in the contractile response of the dog small intestine to opioids. PMID- 2986991 TI - The sympathomimetic activity of (+/-)-pindolol at beta 1- and beta 2-adrenoceptor sites. AB - In isolated right atrial and stilboestrol-pretreated uterine preparations from both guinea-pigs and rats, pindolol elicited propranolol-sensitive positive chronotropic and smooth muscle relaxant actions. Although the pD2 values for pindolol (8.4-9.2) and (-)-isoprenaline (ISO, 8.4-8.7) fell within the same range in these preparations, the maximum responses to pindolol were less than 15% of those to the catecholamine. Thus, pindolol did not display any selectivity for agonistic actions at beta 1- or beta 2-adrenoceptors. In uteri taken from progesterone-pretreated rats, the pD2 value for (-)-isoprenaline was 9.5 and that of pindolol 8.5. In these preparations the maximal relaxant effect of pindolol (approximately 50% Emax ISO) was greater than that found in oestrogen-pretreated uteri. Thus, it appears that the maximal response of pindolol in vitro can be related to the pD2 value for (-)-isoprenaline. In anesthetized cats, intravenous pindolol elicited non-beta-adrenoceptor-mediated increases in heart rate and decreases in soleus muscle contractility. The mechanism(s) underlying the latter actions are unknown. PMID- 2986992 TI - Partial characterization of solubilized platelet imipramine binding sites using a new probe, [3H]3-cyanoimipramine ([3H]Ro 11-2465). AB - Evidence suggests that [3H]imipramine labels the recognition site of the neuronal 5-hydroxytryptamine uptake mechanism. We are investigating the linkage between these binding sites and the carrier by biochemical characterization. [3H]Imipramine-labelled sites have been solubilized from outdated human platelets using the detergent digitonin. [3H]3-Cyanoimipramine binds persistently to these sites in the presence of Na+ at 4 degrees C. At higher temperatures, and in the absence of Na+, this ligand acts reversibly. We report the use of this pseudo irreversible ligand in the initial molecular characterization of the recognition molecule. To confirm that this ligand occupies the [3H]imipramine-labelled sites, human platelets were prelabelled with 3-cyanoimipramine before incubating with [3H]imipramine. Only low affinity [3H]imipramine binding remained. The majority of the 3-cyanoimipramine was irreversibly bound under these conditions as shown by the use of the 3H compound. Gel permeation chromatography of [3H]3 cyanoimipramine-prelabelled platelet membranes solubilized with digitonin indicated a Stokes' radius of 6.3 nm. This is larger than values previously determined for cholate-solubilized sites. We conclude that [3H]3-cyanoimipramine will be useful for further purification and reconstitution studies. PMID- 2986993 TI - Systemic hemodynamic effects of dopamine, (+/-)-dobutamine and the (+)-and (-) enantiomers of dobutamine in anesthetized normotensive rats. AB - The hemodynamic effects of dopamine, (+/-)-dobutamine (racemic mixture) and the (+)- and (-)-enantiomers of dobutamine were evaluated in anesthetized normotensive rats. Dopamine and (+/-)-dobutamine produced hemodynamic effects in anesthetized rat that were qualitatively similar to those reported for these compounds in man. The increase in cardiac output produced by dopamine and (+/-) dobutamine was due mainly to an increase in stroke volume, with heart rate being only minimally affected. Dopamine produced a large increase in mean arterial pressure and slightly increased total peripheral vascular resistance, whereas (+/ )-dobutamine only modestly increased blood pressure and significantly reduced total peripheral resistance. The (-)-enantiomer of dobutamine, which possesses mainly alpha 1-adrenoceptor agonist activity, produced marked increases in cardiac output, stroke volume, total peripheral resistance and mean arterial pressure, but did not significantly increase heart rate. In contrast, (+) dobutamine, which possesses predominantly beta 1-and beta 2-adrenoceptor agonist activity, elicited only a modest increase in cardiac output which was due entirely to increased heart rate since stroke volume was not increased. Total peripheral vascular resistance and mean arterial blood pressure were both reduced by (+)-dobutamine, characteristic of a beta-adrenoceptor agonist. The increase in cardiac output and blood pressure produced by (+/-)-dobutamine, but not the positive chronotropic effect, were significantly inhibited by alpha 1 adrenoceptor blockade with prazosin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2986994 TI - Leukotriene E4 binds specifically to LTD4 receptors in guinea pig lung membranes. AB - High affinity, stereoselective binding sites for [3H]leukotriene E4 ([3H]LTE4) have been identified and characterized in guinea pig lung membranes. [3H]LTE4 bound to these membranes with a pharmacological specificity identical to that previously observed for the [3H]LTD4 receptor in guinea pig lung. [3H]LTE4 specific binding was selectively inhibited by Na+, enhanced by Ca2+, Mg2+ and Mn2+ and modulated by guanine nucleotides. Scatchard analysis of saturation binding data showed a single class of high affinity and saturable binding sites, with a dissociation constant (Kd) of 0.4 +/- 0.2 nM and a density (Bmax) of 430 +/- 50 fmol/mg membrane protein, similar to values observed for the LTD4 receptor in guinea pig lung. The rank order potency of agonist binding to the [3H]LTE4 binding sites was LTD4 greater than LTE4 much greater than LTC4. These results indicate that [3H]LTE4 binds to [3H]LTD4 receptors and suggests that induction of smooth muscle contraction by LTD4 and LTE4 may be mediated by identical mechanisms and receptors in the guinea pig lung. PMID- 2986996 TI - DMSO effects on synaptic facilitation and calcium dependence in bullfrog sympathetic ganglion. AB - Synaptic transmission in the bullfrog sympathetic ganglion was studied by means of extra- and intracellular recordings. DMSO (3-10%) caused a single orthodromic stimulus to generate a brief burst of repetitive postganglionic discharges. DMSO also partially reversed a preexisting transmission failure in low Ca2+ medium. Ganglia were exposed to gradual reductions in extracellular Ca2+, in the absence and in the presence of DMSO. The recorded amplitude of the postganglionic compound action potential (CAP) was plotted as a function of Ca2+ concentration. In the absence of DMSO transmission failed progressively as Ca2+ was reduced from 1.8 to 0.47 mM but DMSO (3% and 10%) shifted the curve of transmission failure to the left (lower Ca2+ concentration). DMSO inhibits ganglion cholinesterase activity, but this is not the mechanism of its facilitatory effect on Ca2+ entry, since physostigmine did not shift the curve of transmission failure in low Ca2+ to the left. The data suggest that DMSO maintained transmitter release in low Ca2+ by a direct, nonspecific enhancement of Ca2+ influx into the presynaptic nerve terminal. PMID- 2986995 TI - Enhancement of brain [3H]flunitrazepam binding and analgesic activity of synthetic cannabimimetics. AB - Novel, synthetic cannabimimetics and delta 9-tetrahydrocannabinol were found to enhance the binding of [3H]flunitrazepam to mouse brain in vivo. This property, suggestive of facilitation of binding to benzodiazepine receptors, is consistent with the potentiation of the anticonvulsant activity of diazepam against pentylenetetrazol by these compounds. The relative potencies of delta 9 tetrahydrocannabinol and the new cannabimimetics for enhancing [3H]flunitrazepam binding in vivo could also be correlated with their relative analgesic efficacies. Similar pharmacological stereospecificity was displayed for both binding enhancement and analgesic effects. The following order of decreasing potency was observed: N-methyllevonantradol and (-)-CP-55,244 greater than levonantradol, canbisol, CP-42,096 and (-)-CP-55,940 greater than 9-beta normethyl-9-beta-hydroxyhexahydrocannabinol, nabilone and CP-47,497 greater than delta 9-tetrahydrocannabinol. Dextronantradol, (+)-CP-55,940 and (+)-CP-55,244 were considerably less active than the respective (-)-enantiomers; cannabidiol was inactive. Extensive investigation of structure versus activity led to N methyllevonantradol and the 3-(2-hydroxyphenyl)cyclohexanols derivative, (-)-CP 55,244, which are approximately 1000-fold more potent than delta 9 tetrahydrocannabinol. PMID- 2986997 TI - The benzodiazepine antagonist Ro 15-1788 reverses the effect of methyl-beta carboline-3-carboxylate but not of harmaline on cerebellar cGMP and motor performance in mice. AB - The cerebellar cGMP level in mice was decreased in a dose-dependent manner 30 min after diazepam (ED50 = 2 mg/kg p.o.). This effect was reversed by the specific benzodiazepine antagonist Ro 15-1788. Methyl-beta-carboline-3-carboxylate (beta CCM) and harmaline increased cGMP. Ro 15-1788 dose dependently counteracted the beta-CCM- but not the harmaline-induced increase in cGMP. In the horizontal wire test Ro 15-1788 antagonized the impairment of motor performance induced by beta CCM, but not that induced by harmaline. These findings further support the view that harmaline in contrast to beta-carboline-3-carboxylates does not act through benzodiazepine receptors, and that Ro 15-1788 antagonizes only those convulsants and stimulants that act through specific benzodiazepine receptors. PMID- 2986998 TI - Effect of vasoactive intestinal polypeptide (VIP) on cyclic AMP level and relaxation in rat isolated aorta. AB - The effects of vasoactive intestinal polypeptide (VIP) on cyclic nucleotide (cAMP and cGMP) levels and smooth muscle relaxation were investigated in rat isolated aorta and compared to those of the beta-adrenergic agonist isoprenaline. VIP increased the cAMP level of rat aorta in a concentration-dependent manner with an EC50 of 0.1 microM. VIP 1 microM maximally increased the cAMP level 7 fold, whereas the beta-adrenergic agonist isoprenaline 10 microM elevated the cAMP level only 2.5 fold. VIP 1 microM relaxed the precontracted rat aorta by only about 16% whereas isoprenaline 10 microM induced a relaxation of about 86%. VIP did not alter the cGMP level and its effect on cAMP content was not changed in the presence of indomethacin (5 microM). No quantitative correlation could thus be established between increases in total tissue levels of cAMP and the degree of relaxation in rat isolated aorta. PMID- 2986999 TI - [3H]U-69593 a highly selective ligand for the opioid kappa receptor. AB - The selective kappa agonist U-50488 was recently discovered and characterized. In this study, the receptor binding properties of [3H]U-69593, an analog of U-50488, were characterized. [3H]U-69593 binds with high affinity (3 nM) to membranes prepared from guinea pig, mouse and rat brain. The number of kappa binding sites comprise only 13%, 9% and 4% of the total opioid sites, respectively. The benzmorphans, dynorphin, and compounds structurally related to U-50488 have high affinity for this kappa site. PMID- 2987000 TI - Identification of subtypes of [3H]prazosin-labelled alpha 1 receptor binding sites in rat brain. AB - Detailed antagonist competition curves for [3H]prazosin-labelled binding sites in rat cerebral cortex membranes reveal anomalous binding characteristics. Dihydroergocryptine and indoramine compete in a steep, monophasic manner while WB4101 and phentolamine exhibit shallow competition curves. Computer-assisted analysis of binding data indicate that both WB4101 and phentolamine discriminate identical subpopulations of [3H]prazosin binding sites, which each comprise approximately 50% of specific [3H]prazosin binding. These data suggest the presence of subtypes of [3H]prazosin-labelled alpha 1 adrenergic receptors in rat cerebral cortex. PMID- 2987001 TI - Neuropeptide Y (NPY): enhancement of blood pressure increase upon alpha adrenoceptor activation and direct pressor effects in pithed rats. AB - The effects of neuropeptide Y (NPY) on blood pressure and heart rate were studied in pithed rats. Systemic infusion of NPY in a dose (230 pmol X kg-1 X min-1) which per se did not affect blood pressure enhanced the pressor response to phenylephrine (50 nmol X kg-1 i.v.) and that to electrical stimulation of the sympathetic outflow. In higher doses, NPY caused a pressor effect per se, which was dose-dependently antagonized by nifedipine but not by adrenoceptor antagonists. In conclusion, NPY enhanced the alpha-adrenoceptor-mediated response and had Ca2+-dependent vasoconstrictor activity in vivo. PMID- 2987002 TI - Receptor interactions in stimulation of hydrolysis of inositol phospholipids. AB - The amount of carbachol-induced hydrolysis of inositol phospholipid in the mouse exocrine pancreas is reduced when cholecystokinin octapeptide, an agonist which works through a separate class of receptors, is also present. This effect does not appear to be due to depletion of inositol phospholipid from the cell membranes. These results indicate that the hydrolysis of inositol phospholipid is not always dependent on the number of receptors occupied and that the different classes of receptors share at least one component in the response system. PMID- 2987003 TI - Ontogenetic properties of benzodiazepine receptor subtypes in rat spinal cord. PMID- 2987004 TI - Neuropeptide Y (NPY): an endogenous modulator of non-adrenergic transmission in rat vas deferens? PMID- 2987005 TI - DNA topoisomerase I and II activities during cell proliferation and the cell cycle in cultured mouse embryo fibroblast (C3H 10T1/2) cells. AB - We have used C3H 10T1/2 cells to examine the regulation of topoisomerase activities during cell proliferation and the cell cycle. The specific activity of topoisomerase I was about 4-fold greater in proliferating (log phase) cells than in non-proliferating (confluent) cells. In synchronized cells, the bulk of the increased activity occurred during or just prior to S phase, depending upon the method of synchronization. A smaller increase in activity also occurred during G1 phase. The increase in activity during S phase was not altered by a hydroxyurea block at the G1/S phase boundary indicating that it is not directly coupled to DNA synthesis and is not the result of topoisomerase I gene dosage. The increase was inhibited by blocking cells at mid-G1 phase using isoleucine deprivation. Thus, the increase in activity during S phase is dependent on events occurring during mid- to late G1 phase. In contrast to the changes in topoisomerase I levels, the specific activity of topoisomerase II showed no detectable difference in proliferating vs non-proliferating cells. In addition, no detectable difference in topoisomerase II specific activity was seen in G1, S and M phases of the cell cycle. The differences in the activity profiles of the topoisomerases I and II during the cell cycle suggest that the two activities are regulated independently and may be required for different functions. PMID- 2987006 TI - Further characterization of the inhibition of casein production in a primary mouse mammary epithelial cell culture by epidermal growth factor. Antagonism by cyclic AMP. AB - Epidermal growth factor (EGF) inhibited casein production and the accumulation of casein mRNA activity induced by insulin (I), cortisol (F) and prolactin (P) in a primary culture of mammary epithelial cells from pregnant mice. The inhibitory effects of EGF were blocked by 8-bromo cyclic AMP (8-br-cAMP) in a dose-dependent manner. The effect of 8-br-cAMP was observed at a concentration as low as 20 microM and was maximal at 500 microM. Dibutyryl cyclic AMP (db-cAMP), cAMP, and 3 isobutylmethylxanthine (IBMX), an inhibitor of phosphodiesterase, also antagonized the inhibitory effect of EGF on casein production. 8-Br-cAMP had, however, no effect on the mitogenic activity of EGF in this system. These results suggest a possible modulatory role of cAMP in EGF-induced inhibition of casein production in cultured mammary epithelial cells. PMID- 2987007 TI - Establishment and characterization of a permanent pSV ori--transformed ataxia telangiectasia cell line. AB - A permanent ataxia-telangiectasia (A-T) cell line has been established from the fibroblast strain AT2SF after transfection with the bacterial plasmid pSV ori-, which contains replication origin-defective SV40 sequences. The original transfection frequency, as measured by transformed foci, was markedly reduced in two A-T strains when compared with either normal or xeroderma pigmentosum fibroblasts. As with SV40 virion-transformed fibroblasts, pSV ori--transformed cells entered a crisis phase, from which about one-fourth of the original clones from A-T and normal fibroblasts recovered. Both the pSV ori--transformed TAT2SF cell line and an SV40 virion-transformed AT5BI (GM5489) cell line retained their characteristic sensitivity to the lethal effects of ionizing radiation, as well as their X ray-resistant DNA synthesis. Southern blot analysis of cellular SV40 sequences demonstrated a single major integration site of pSV ori- in the AT2SF cells. In contrast, AT5BI cells transformed with SV40 virions demonstrated a high degree of heterogeneity of integrated viral sequences. Neither the TAT2SF nor the GM5489 transformed cell line contains any detectable freely replicating SV40 viral sequences, which are seen in many other semipermissive SV40-transformed cells. PMID- 2987008 TI - Antibodies to nuclear thyroid hormone-binding proteins. Antibody characterization and immunofluorescent localization. AB - To further investigate the mechanism by which thyroid hormones regulate target cell function, we have prepared and partially characterized antibodies to highly purified nuclear thyroid hormone-binding proteins (NTBP). NTBPs were prepared from bovine liver nuclear extracts by bio-specific elution from an affinity gel containing immobilized 3,5,3'-triiodo-L-thyronine (T3). Antibodies (Ab) raised to NTBP in BALB/c mice were assayed for Ab-NTBP complex formation on HPLC TSK SW3000 molecular exclusion gels and found to be species-specific and non-cross-reactive with serum thyroid hormone-binding proteins. Most of the antibody activity was directed against two fractions of molecular weight (MW) 89 000 and 53 000, which were associated with thyroxine (T4)-binding activity. The 89 000 D T4-binding activity was shifted to a higher MW complex when incubated with specific antibody. Indirect immunofluorescence showed antibody activity against discrete, clumped chromatin sites, nuclear envelope and plasma membrane in hepatocytes. Intense fluorescence was also observed in the cells lining the hepatic sinusoids and in the cytoskeleton of bovine aortic endothelial cells in culture. The data suggest that thyroid hormone target cells contain extranuclear loci that share antigenic sites with NTBP and may also represent specific NTBP-like sites of thyroid hormone binding. PMID- 2987009 TI - Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture. AB - Mouse L fibroblasts and other mammalian cells are killed by the translation inhibitor hygromycin B. We have modified the gene conferring resistance against hygromycin B in E. coli in such a way that it can be transcribed in mammalian cells from the promoter of the HSVtk gene. The resulting plasmid, pHMR272, was transfected into mouse L fibroblasts and HeLa cells by the calcium phosphate method and upon selection produced clones resistant against hygromycin B. The transfection rate was similar to that obtained with other selective markers. This plasmid is a useful addition to the relatively small number of dominant selectable markers available for mammalian cells. PMID- 2987010 TI - Mononuclear cell-mediated modulation of synovial cell metabolism. I. Collagen synthesis. AB - Human PHA-stimulated mononuclear cells produce a factor which inhibits synovial cell collagen and non-collagen protein synthesis, whereas it enhances hyaluronic acid (HA) production. Indomethacin (10(-4)-10(-6) M), a cyclo-oxygenase inhibitor, suppresses this effect, suggesting that the mechanism is prostaglandin mediated. The active material, of apparent molecular weight 12 000-20 000, also displays the properties of the mononuclear cell factor (MCF) previously described by others, since its stimulates collagenase and PGE2 release by the cultured synovial cells. Furthermore, it co-purifies with interleukin 1 (IL 1) as shown by lymphocyte-activating factor activity. This strongly suggests that IL 1 could be responsible for some (or all) the effects observed on MCF-exposed synovial cells. From these data, we deduce the possibility that mononuclear cells may participate in limiting synovial collagen deposition in rheumatoid arthritis. PMID- 2987011 TI - Changes in 5'-nucleotidase and adenosine deaminase distribution across the rat left ventricle wall during growth and aging. AB - The concentrations of the adenosine-generating enzyme 5'-nucleotidase (5'-N) and of the adenosine-degrading enzyme adenosine deaminase (ADA) in the rat left ventricle change as a function of the age of the animal. The enzyme distribution across the left ventricle wall is non-uniform in adult or old rats (in the case of 5'-N) or in all age-groups (in the case of ADA). In the oldest rats, 5'-N activity exhibited a significant increase in the mid-myocardium and in the inner myocardial layers as compared with the young adult controls. PMID- 2987013 TI - Control of locomotion in marine mollusc Clione limacina. II. Rhythmic neurons of pedal ganglia. AB - Activity from neurons in isolated pedal ganglia of Clione limacina was recorded intracellularly during generation of rhythmic swimming. To map the distribution of cells in a ganglion, one of two microelectrodes was used to monitor activity of the identified neuron (1A or 2A), while the second electrode was used to penetrate successively all the visible neurons within a definite area of the ganglion. In addition, pairs of neurons of various types were recorded in different combinations with each other. Intracellular staining of neurons was also performed. Each ganglion contained about 400 neurons, of which about 60 neurons exhibited rhythmic activity related to a swim cycle. These rhythmic neurons were divided into 9 groups (types) according to axonal projections, electrical properties and the phase of activity in a swim cycle. Three types of interneurons and six types of efferent neurons were distinguished. Type 7 and 8 interneurons generated only one spike of long (50-150 ms) duration per swim cycle. Type 7 interneurons discharged in the phase of the cycle that corresponded (in actual swimming) to the dorsal movement of wings (D-phase). Type 8 interneurons discharged in the opposite phase corresponding to the ventral movement of wings (V-phase). With excitation of type 7 interneurons, an IPSP appeared in the type 8 interneurons, and vice versa. Neuropilar branching of these neurons was observed in the ipsilateral ganglion. In addition, they sent an axon to the contralateral ganglion across the pedal commissure. Efferent neurons (i.e. the cells sending axons into the wing nerve) generated spikes of 1-5 ms duration. Type 1 and 3 neurons were excited in the D-phase of a swim cycle and were inhibited in the V-phase. Type 2 and 4 neurons were excited in the V-phase and inhibited in the D-phase. Type 10 neurons received only an excitatory input in the V-phase, while type 6 neurons received only an inhibitory input in the D phase. Type 12 interneurons were non-spiking cells, they generated a stable depolarization ("plateau") throughout most of the V-phase. Neurons of the same type from one ganglion (except for type 6) were electrically coupled to each other. There were also electrical connections between most neurons firing in the same phase of the cycle, i.e. between types 3 and 7, as well as between types 2, 4 and 8. Type 7 interneurons from the left and right ganglia were electrically coupled, the same was true for type 8 interneurons. PMID- 2987014 TI - Control of locomotion in marine mollusc Clione limacina. IV. Role of type 12 interneurons. AB - Type 12 interneurons in pedal ganglia of Clione limacina exerted a strong influence upon the locomotor generator during "intense" swimming. These neurons generated "plateau" potentials, i.e. their membrane potential had two stable states: the "upper" one when a neuron was depolarized, and the "down" one, separated by 30-40 mV. The interneurons could remain in each state for a long time. Short depolarizing and hyperpolarizing current pulses, as well as excitatory and inhibitory postsynaptic potentials, could transfer the interneurons from one state to another. When the pedal ganglia generated the locomotory rhythm, type 12 neurons received an EPSP and passed to the "upper" state in the V2-phase of a locomotor cycle. They remained at this state until the beginning of the D1-phase when they received an IPSP and passed to the "down" state. The EPSP in type 12 neurons was produced by type 8d neurons, and the IPSP by type 7 neurons. Type 12 neurons exerted inhibitory influences upon many neurons active in the V1 and V2 phases, and excitatory influences upon the D phase interneurons (type 7). The functional role of type 12 neurons was to limit the activity of neurons discharging in the V-phase of a locomotory cycle. In addition, they enhanced the excitation of the D-phase neurons and promoted, thus, the transition from the V-phase to the D-phase. PMID- 2987012 TI - Descending control of spinal nociceptive transmission. Actions produced on spinal multireceptive neurones from the nuclei locus coeruleus (LC) and raphe magnus (NRM). AB - The effects of electrical stimulation in the nuclei locus coeruleus (LC) and raphe magnus (NRM) were examined on the background and/or evoked discharge of neurones in the spinal dorsal horn of anaesthetized cats. These were qualitatively, and in most cases quantitatively similar, in their action on multireceptive neurones. In these neurones an inhibitory action on the discharge evoked by noxious cutaneous stimuli or by activation of A delta and C fibres was most prominent although in some neurones (22%) an initial excitation lasting up to 100 ms preceded the inhibition which could last up to 1 s. Excitation alone was observed in only 3% of multireceptive neurones. Electrical stimulation also produced an inhibitory action on the discharge of low threshold mechanoreceptive neurones (80%). In four of ten multireceptive neurones examined in detail, LC stimulation produced a selective inhibitory action on the discharge evoked by noxious cutaneous stimuli. In the remaining six multireceptive neurones it was partially selective against noxious as compared with non-noxious inputs. The inhibitory action was also more pronounced on the discharge evoked by activity in A delta and C fibres than fast conducting afferents. The inhibitory action evoked by electrical stimulation in LC on nociceptive transmission in the spinal cord is suggested to play a part in mediating analgesia from LC. PMID- 2987015 TI - The differential effects of baclofen on segmental and descending excitation of spinal interneurones in the cat. AB - Intravenous baclofen (1-6.25 mg kg-1) substantially reduced the monosynaptic excitation of neurones in the intermediate nucleus of the cat spinal cord by impulses in group I extensor muscle primary afferent fibres, but had little or no effect on excitation by stimulating fibres of the ipsilateral dorsolateral funiculus or the contralateral red nucleus. Relatively low concentrations of baclofen thus appear not to influence the release of excitatory transmitter from the terminals of rubrospinal, corticospinal and long descending propriospinal fibres, in contrast to the reduction of the release of primary afferent transmitters. PMID- 2987016 TI - Electromechanical changes during electrically induced and maximal voluntary contractions: surface and intramuscular EMG responses during sustained maximal voluntary contraction. AB - Changes in the electrical activity of the human gastrocnemius and soleus muscles during fatiguing maximal plantar flexions were studied with computer-aided EMG frequency power spectral analysis and intramuscular spike amplitude-frequency histogram analysis. In some experiments, brief supramaximal nerve stimulations of 80 Hz were given at 15-s intervals during sustained maximal voluntary contractions (MVCs). Multiple muscle biopsy samples were also obtained from the gastrocnemius muscle for fiber type determination. The surface EMG frequency spectral analysis showed a highly significant reduction in mean power frequency and root mean square EMG amplitude during sustained MVCs. The intramuscular spike amplitude-frequency histograms showed that the gastrocnemius muscle had a progressive reduction in the motor unit discharge frequency, particularly those with a relatively high amplitude, whereas the soleus muscle hardly showed a reduction in motor unit activity. Reduction in motor unit activity was also found to be more pronounced in gastrocnemius muscles with higher proportions of type II fibers. Brief maximal tetanic stimulations initially matching the MVC failed to increase the contraction force. Similarly, the evoked compound mass action potentials showed little change in the amplitude in subjects with different muscle fiber compositions. Results of this study suggest that during sustained MVCs, force fatigue could not be attributed to a failure of muscle membrane electrical propagation; a progressive reduction in motor unit activation does not result in a functional disadvantage, but may optimize excitation-contraction coupling by avoiding a muscle electrical conduction failure; and the extent of the reduction in motor unit activation seems to be muscle-fiber-type-dependent which may account for the reduction in amplitude and frequency of the surface EMG. PMID- 2987017 TI - Actions of kynurenic acid and quinolinic acid in the rat hippocampus in vivo. AB - An iontophoretic study was made of the interaction of kynurenic acid with excitatory amino acids in the hippocampus and with the commissural input from the contralateral hippocampus in the rat. The results showed that kynurenic acid was an effective blocker of synaptic transmission in the hippocampus in vivo, adding further support to the idea that an excitatory amino acid is involved in neurotransmission in this structure. In addition there was an increase in the specificity of kynurenate as an antagonist of excitatory amino acids in the hippocampus compared with neocortex, with much more activity being shown toward the NMDA-preferring rather than the quisqualic acid-preferring receptor. Kynurenic acid was also able to distinguish partially between quinolinic acid and NMDA, supporting the possibility that two types of NMDA/quinolinate receptors exist in the hippocampus. PMID- 2987018 TI - UDPgalactose:ceramide galactosyltransferase and 2',3'-cyclic-nucleotide 3' phosphodiesterase activities in rat brain after long-term exposure to methylmercury or triethyllead. AB - Methylmercury (MeHg) and triethyllead (Et3Pb) are known to cause neurologic impairment in human and in several animal models. In the developing central nervous system the formation of myelin is particularly vulnerable. To obtain more information on the toxic mechanisms related to dysmyelination, the effects of MeHg and Et3Pb on two marker enzymes of myelination was assessed in developing rats. From the 5th day of life intraperitoneal injections of MeHgCl or Et3PbCl at doses of 0.05 to 5 mg/kg body weight were administered to the rats three times a week. They were decapitated at the 21 to 23rd (group A) or at the 28 to 31st postnatal day (group B). The animals treated with 2 mg/kg MeHg or Et3Pb appeared normal and the rate of growth was unchanged compared with that of control rats. A decreased activity of the enzymes UDP galactose:ceramide galactosyltransferase (CGalT) and 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNP) was apparent already at doses of 0.1 mg/kg in group B rats. (MeHg, 18 and 16%, respectively; Et3Pb, 11 and 14%) and the values decreased further with increased toxic doses. In the MeHg-treated animals the exposure time was decisive for the effect; thus in group A of MeHg-treated animals the change in enzyme activities was minimal at doses which in group B had an inhibitor effect. The activities of brain acetylcholinesterase and succinate dehydrogenase were not affected. The results emphasize a common early effect of MeHg and Et3Pb on enzymes associated with myelination in the developing central nervous system. PMID- 2987019 TI - A polypeptide immunoreactive nonchromaffin paraganglioma in the periglandular connective tissue of glandula suprarenalis of a dog. A case report. AB - A nonchromaffin paraganglioma was found in the periglandular connective tissue of the glandula suprarenalis of a sheep-dog bastard and characterized by histological and immunohistochemical techniques. Insulin-, glucagon- and gastrin/cholecystokinin-like material was revealed within the cells of the tumour. Possible functional consequences of these findings are discussed. PMID- 2987020 TI - Modulation of protein kinases and phosphoprotein phosphatases by a small acidic protein from bovine brains. AB - A small, acidic and heat-stable protein was purified from bovine brains by column chromatography on DEAE-cellulose, Bio-Gel HTP, Affi-Gel phenothiazine and Sephadex G-75. This protein stimulates megamodulin-dependent protein kinase I from brains and phosphoprotein phosphatases from either brain or yeast. However, it inhibits cyclic AMP-dependent protein kinases from skeletal muscle. PMID- 2987021 TI - D-ala2-Metenkephalinamide blocks the synaptically elicited cortical spreading depression in rats. AB - Spreading depression (SD) was elicited in rats anesthetized with pentobarbital by a train of 8 electrical pulses (0.1 ms, 10 Hz) applied to parietal cortex. Local application of 50 micrograms of D-ala2-metenkephalinamide (DAME) on the stimulated area evoked one or two SD waves followed by an increase of SD threshold from 40 V to 90 V. This effect could be partly prevented by naloxone (1 mg/kg i.p.) and reversed by local application of 4-aminopyridine (10(-3) M, 2 microliters), which reduced SD threshold to 5 and 20 V in normal and DAME-treated cortex, respectively. It is argued that DAME exerts an inhibitory effect on cortical neurons and that the initial SD facilitation is due to initial blockade of inhibitory neurons in the superficial cortical layers. PMID- 2987023 TI - In vivo and in vitro studies on the release of cortisol from interrenal tissue in trout. I. Effects of ACTH and prostaglandins. AB - The secretion of cortisol from the interrenal tissue in trout Salmo gairdnerii was studied in vivo and by an in vitro superfusion method in relation to the effect of adrenocorticotropic hormone (ACTH), prostaglandins (PGE1, PGF2 alpha) and prostaglandin inhibitors. The experiments were performed between April and July. In control, uninjected animals, circulating cortisol was 206 +/- 24 ng/ml. At 24 h following the intraperitoneal injection of indomethacin, dexamethasone, or both, the levels were reduced to 84 +/- 10, 43 +/- 20 and 63 +/- 11 ng/ml respectively. In control animals receiving solvent (ethanol), the value was 190 +/- 30. With longer term duration (72 h) the levels of plasma cortisol following injection of indomethacin, dexamethasone and both drugs together were 120 +/- 29, 120 +/- 10 and 37 +/- 8 respectively. On the contrary, no significant change was produced by prostaglandins. In vitro release of cortisol from superfused head kidneys (which contain the interrenal tissue of teleosts) decreased gradually with time and reached a minimum after 50 min. Addition of ACTH or of PGE1 to the medium induced an immediate, dose dependent increase in cortisol output which reached a maximum after 15-30 min. On the contrary, PGE2 alpha or indomethacin did not modify cortisol release. These results show that cortisol production in trout, which is controlled by ACTH as in other vertebrates, is also under the influence of other hormonal substances acting on the adenyl-cyclase system. PMID- 2987022 TI - Role of phospholipids in the binding activity of vasoactive intestinal peptide receptors. AB - Phospholipase digestion of rat intestinal epithelial cell membranes was performed in order to study the influence of membrane phospholipids on the binding activity of VIP receptors. Phospholipases A2 and C strongly (ED50 congruent to 4 X 10(-2) and 4 X 10(-1) micrograms/ml, respectively) and rapidly reduced 125I-VIP binding to membranes whereas phospholipase D was ineffective. This suggests an important role of both hydrophobic and hydrophilic groups of phospholipids on VIP receptor binding activity. PMID- 2987024 TI - Transplantable chemically-induced liver tumors in the viviparous fish Poeciliopsis. AB - Transplantation of five liver tumors induced with the chemicals diethylnitrosamine (DEN) and 7,12-dimethylbenz[a]anthracene (DMBA) in small live bearing fish of the genus Poeciliopsis is reported. Five permanent strains representing three distinct tumor types were established in isogenic hosts. Histological characterization of hepatocellular carcinoma, hemangiopericytic sarcoma, and cholangiocarcinoma and the development of the neoplasms in host fish is presented. Transplantability of the experimental liver tumors provides evidence of their malignant nature. Metastasis of the hepatocellular carcinoma occurred from tumor implants in the dorsal musculature or peritoneal cavity and from the hemangiopericytic sarcoma implanted in the intraperitoneal cavity. PMID- 2987025 TI - A comparison of the effects of cytotoxic cerebrospinal fluid on cell cultures with other cytopathogenic agents. AB - Protein synthesis, antigen synthesis, and cell membrane permeability were analyzed after inoculating human diploid fibroblasts with control or cytotoxic CSF, herpes simplex virus type 1 (HSV 1), poliovirus 3, or Clostridium difficile toxin. Whereas protein synthesis and membrane permeability were affected by the viruses, and virus antigens detectable by pooled human serum were synthesized, the bacterial toxin and cytotoxic CSF did not induce any new proteins or antigens, although the cytotoxic CSF reduced cellular protein synthesis levels and caused an increase in the permeability of the cell membranes. The effect of the cytotoxic CSF in cell culture resembles that of a toxin rather than a replicating virus. PMID- 2987027 TI - [Possibilities for the external regulation of lysosomal structure and functions. Pharmacological effects on the permeability of lysosomal membranes in the ischemic myocardium]. PMID- 2987026 TI - [Effect of benzonal on the liver mono-oxygenase system of laboratory animals]. AB - The data are provided on benzonal as having the properties of the monoxygenase system inducer, evidenced by the shortening of hexenal sleep, increase of the intensity of the epr-signal of cytochrome P-450, activation of lipid peroxidation, carboxylesterase and arylesterase in the liver. The properties of benzonal described can be made use of for increasing the body resistance to the toxic action of xenobiotics. PMID- 2987028 TI - [Antenatal effect of imipramine on the ontogeny of the imipramine receptors in the cerebral cortex of rats]. AB - Ontogenetic study of imipramine binding sites in the rat brain cortex employing autoradiographic and radioreceptor methods revealed that they can be detected at the 19th day of gestation and that by the 14 day of postnatal development their amount reaches the adult levels. The affinity constants of imipramine binding sites did not change significantly throughout the ontogenetic development. Exposure to the therapeutic doses of imipramine on days 17-19 of gestation resulted in a significant increase in the amount of binding sites (24% of the control level) on day 3 of postnatal development, which returned to normal by the end of the second week of postnatal development. The distribution of imipramine binding sites in the cortical layers did not change either in normal rats or in those antenatally treated with imipramine. PMID- 2987029 TI - [Tetrodotoxin competition with muscimol for the gamma-aminobutyric acid receptor]. AB - It has been shown that tetrodotoxin inhibits GABA-receptor binding of muscimol with solubilized membranes of the rat brain in a competitive manner, with the inhibition constant amounting to about 3 nM. Muscimol has been also demonstrated to competitively inhibit 3H-tetrodotoxin binding with the same drug. PMID- 2987030 TI - [Effect of the neuropeptide tuftsin and its derivatives on metabolism in various structures of the brain]. AB - A study was made of the effect of the neuropeptide tuftsin and its derivatives on some indicators of the metabolism in limbic structures and cortex of the brain in 200 rats. It was shown that tuftsin and its derivatives raise the content of nicotinamide coenzymes at the expense of the oxidized forms of NAD, activate cytochrome-c-oxidase and dehydrogenases largely in the limbic structures. Thus the peptides under study were found to increase the intensity of cell respiration and to enhance intracellular redox processes, i. e. to largely activate the vital activity of the cells of the brain emotiogenous structures. PMID- 2987031 TI - 13C-NMR studies of horse ferrocytochrome c. Assignment and temperature dependence of methyl resonances. AB - The 13C and proton chemical shifts of 53 of the 55 methyl resonances of horse ferrocytochrome c have been determined by editing natural abundance 13C spectra according to the number of attached protons, observing the temperature dependence of the chemical shifts, and correlating 13C and proton chemical shifts in two dimensional spectra. Previous assignments of proton shifts allow 16 of the 13C resonances to be assigned firmly. PMID- 2987032 TI - Replacement of the deoxycytidine residues in Rhizobium bacteriophage RL38JI DNA. AB - Rhizobium phage RL38JI DNA is resistant to cleavage by a variety of restriction endonucleases, and is only partially sensitive to digestion by pancreatic DNase I or by micrococcal nuclease. We have found that a mixture of DNase I, P1 nuclease, and bacterial alkaline phosphatase will quantitatively digest RL38JI DNA to deoxyribonucleosides. HPLC analysis revealed that dCyd is nearly totally absent among these digestion products, while dGuo, dAdo, and Thd are readily detected. Three additional peaks are always present; their retention properties correspond to no known modified deoxyribonucleosides. Thus it appears that dCyd is replaced in phage RL38JI DNA by as many as 3 different modified residues. PMID- 2987033 TI - Cyclic AMP as an intraspheroidal differentiation signal in Volvox carteri. AB - The action of the macromolecular inducer glycoprotein on sexual reproduction in the green alga Volvox carteri can be modified by altering the external (intraspheroidal) cAMP concentration. Direct proof for the presence of cAMP in the spheroids is given. Protein binding assay and HPLC-fluorimetric analysis independently demonstrate the existence of cAMP in the matrix, cells, and culture medium. Its concentration is higher in sexual cultures, pointing to a transmitting function in sex induction. The presence in the matrix of other members of a protein phosphorylation system suggests an induction-specific signal cascade in this plant. PMID- 2987034 TI - An activator of protein kinase C (phorbol-12-myristate-13-acetate) augments 2 chloroadenosine-elicited accumulation of cyclic AMP in guinea pig cerebral cortical particulate preparations. AB - Norepinephrine and histamine markedly augment accumulations of cyclic AMP elicited by 2-chloroadenosine in a guinea pig cerebral cortical vesicular preparation. In addition, these biogenic amines stimulate phosphatidylinositol turnover. Phosphatidylinositol turnover is associated with mobilization of internal calcium and with stimulation of protein kinase C. Phorbol-12-myristate 13-acetate (PMA), a known activator of protein kinase C, has no effect on cyclic AMP levels alone, but in a concentration-dependent manner enhances accumulations of cyclic AMP elicited by 2-chloroadenosine. PMA, like norepinephrine, also enhances accumulations of cyclic AMP elicited by histamine. PMA has no effect on the synergistic accumulations of cyclic AMP elicited by combinations of amines and 2-chloroadenosine. PMA also augments accumulations of cyclic AMP elicited by forskolin. The results suggest that activation of phosphatidylinositol turnover by biogenic amines may lead via stimulation of protein kinase C to enhanced responsiveness of cyclic AMP-generating systems. PMID- 2987035 TI - Pentose cycle pathway in normal and tumoral islet cells. AB - Relative to protein content, the activity of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase and the rate of glucose metabolism by the pentose cycle pathway in tumoral insulin-producing cells were similar to or higher than those found in normal rat islets. Hence, the decreased secretory response of tumoral cells to glucose is apparently not attributable to any major anomaly in glucose handling by the hexose monophosphate pathway. PMID- 2987036 TI - Relaxation of PvuII recognition sequence. AB - The substrate specificity of PvuII endonuclease is relaxed in the presence of dimethyl sulfoxide. The new recognition sequences cleaved in pBR322 DNA have been found to be CCGCTG, CATCTG, CAGATG, CAGGTG and CAGCGG. PMID- 2987037 TI - Calcium-activated, phospholipid-dependent protein kinase activity and protein phosphorylation in HL60 cells induced to differentiate by retinoic acid. AB - Treatment of human promyelocytic (HL60) cells with retinoic acid for at least 48 h causes differentiation to more mature myeloid forms. Prior to commitment of cells to the myeloid pathway there is a marked increase in cytosolic calcium activated, phospholipid-dependent protein kinase activity. This increase does not result from an intracellular redistribution of the enzyme. Concomitant with the increased enzyme activity there is enhanced phospholipid-dependent phosphorylation of proteins of 29, 49, 52, 58, 68, 69, 120, 170, 200 and 245 kDa. PMID- 2987038 TI - Superoxide dismutase inhibits the superoxide-driven Fenton reaction at two different levels. Implications for a wider protective role. AB - Superoxide dismutase (SOD) completely inhibits the damage caused by a ferric-EDTA chelate in the presence of a superoxide-generating system. In this reaction superoxide is enzymically dismuted to hydrogen peroxide. Since hydrogen peroxide and a ferric-EDTA chelate are themselves a hydroxyl radical-generating system, it follows that SOD must also protect against damage done by this reaction. The ability of SOD to inhibit damage to deoxyribose caused by hydrogen peroxide and a ferric-EDTA chelate is experimentally demonstrated in this paper. PMID- 2987039 TI - Superoxide generation by either 1-oleoyl-2-acetylglycerol or A23187 in human neutrophils is enhanced by indomethacin. AB - Indomethacin at a concentration (10(-4) M) which depressed the effect on O2- generation by fMet-Leu-Phe, markedly enhanced O-2 generation by both 1-oleoyl-2 acetylglycerol and the calcium ionophore, A23187. These results are explicable in terms of the hypothesis that synergism between cytosolic calcium and protein kinase C is involved in signal transduction for the respiratory burst in the human neutrophil. PMID- 2987040 TI - Modulation of gastric H+,K+-transporting ATPase function by sodium. AB - Gastric H+,K+-ATPase activity is not affected by Na+ at pH 7.0 but is significantly stimulated by Na+ at pH 8.5. For the stimulation at the latter pH, the presence of both Na+ and K+ were essential. Contrary the H+,K+-ATPase, the associated K+-pNPPase was inhibited by Na+ at both pH values. Sodium competes with K+ for the K+-pNPPase reaction. Also, unlike the H+, K+-ATPase activity the ATPase-mediated transport of H+ within the gastric microsomal vesicles was inhibited by Na+. For the latter event only the extravesicular and not the intravesicular Na+ was effective. The data suggest that the K+-pNPPase activity does not represent the phosphatase step of the H+,K+-ATPase reaction. In addition, the observed inhibition of vesicular H+ uptake by Na+ appears to be due to the displacement by Na+ of a cytosolic (extravesicular) H+ site responsible for the vectorial translocation of H+. PMID- 2987041 TI - Murine mammary FM3A carcinoma cells transformed with the herpes simplex virus type 1 thymidine kinase gene are highly sensitive to the growth-inhibitory properties of (E)-5-(2-bromovinyl)-2'-deoxyuridine and related compounds. AB - Murine mammary carcinoma (FM3A TK-/HSV-1 TK+) cells, which are thymidine kinase (TK)-deficient but have been transformed with the herpes simplex virus type 1 (HSV-1) TK gene are inhibited in their growth by (E)-5-(2-bromovinyl)-2' deoxyuridine (BVDU), (E)-5-(2-iodovinyl)-2'-deoxyuridine (IVDU) and (E)-5-(2 bromovinyl)-2'-deoxycytidine (BVDC) at 0.5, 0.5 and 0.8 ng/ml, respectively; i.e., a concentration 5000 to 20 000-fold lower than that required to inhibit the growth of the corresponding wild-type FM3A/0 cells. Hence, transformation of tumor cells with the HSV-1 TK gene makes them particularly sensitive to the cytostatic action of BVDU and related compounds. PMID- 2987042 TI - The chromone inhibitor stigmatellin--binding to the ubiquinol oxidation center at the C-side of the mitochondrial membrane. AB - Stigmatellin, a chromone inhibitor acting at the Q0 center of the bc1 complex, binds to the heme b-566 domain of cytochrome b as well as to the Fe2S2 protein. Its binding induces a shift of the alpha-band of heme b-566 to 568 nm. It does not influence the ligand field of the heme b-562 center. Concomitant with the red shift, stigmatellin gives rise to an alteration of the EPR line shape of the Fe2S2 cluster, namely linewidth narrowing and g value shifts at all 3 principal values. The midpoint redox potential of the Fe2S2 protein is shifted from 290 to 540 mV. PMID- 2987043 TI - In vitro and in vivo evaluation of latex condoms using a two-phase nonoxynol 9 system. AB - In vitro studies were carried out that indicated that a lubricant system consisting of 0.45 +/- 0.1 ml of silicon fluid containing 6.6% +/- 0.5% by volume of nonoxynol 9 and a spermicidal cream consisting of 0.45 +/- 0.1 ml made up of 63.4% polyethylene glycol 400 and 30.0% polyethylene glycol 3350 containing 6.6% +/- 0.5% nonoxynol 9 was effective in reducing sperm motility and viability. This system was tested in vivo with the use of simulated rupture techniques and was found to be equally as effective. Double-blind preference studies were carried out in vivo which showed that the condom system is convenient and comfortable to use, nonirritating to the vagina or urethral mucosa, and esthetically pleasing to the young, reproductive-age population. PMID- 2987045 TI - [Effect of gamma-aminobutyric acid, taurine and catecholamines on the synaptic processes in Lorenzini's ampules in Black Sea skates]. AB - Perfusion of the basal membrane of the ampullae of Lorenzini with GABA, taurine or catecholamines produced a reversible decrease in resting and evoked activity in afferent nerve fibers. Picrotoxine, phentolamine and propranolol did not suppress the electrical activity. Our findings exclude GABA, taurine and catecholamines as the transmitters at the ampullary electro-receptorsensory nerve synapses. PMID- 2987044 TI - [Effects of a salt mixture on the state of the mother, fetus and newborn in experimental hypoxia]. PMID- 2987047 TI - [Analysis of the factors affecting cardiac output in exclusion of the sino carotid reflexogenic area]. AB - In acute experiments on cats, the character and mechanisms of changes of the heart pumping function were studied in suppression of the carotid sinus baroreceptors with occlusion of the carotid arteries. In 65% of tests the cardiac output was reduced. Changes of venous return, at that, were diverse and opposite to those of the cardiac output in 50% of observations. Neither vagotomy, nor blockade of beta-adreno- and M-cholinoreceptors affected the character of cardiac output changes. The role of maximal responses of arterial pressure in forming of the cardiac output shifts in sino-carotid reflex, is discussed. PMID- 2987046 TI - [Effect of hydrocortisone on neuromuscular transmission in the frog skeletal muscles]. AB - Hydrocortisone (1.10(-7)-1.10(-5) M/l) in vitro experiments on frog's muscle in conditions of preliminary curarization increases dose-dependently the quantum content of e.p.p., affecting neither membrane potential, nor input resistance, nor sensitivity to acetylcholine of the membrane of m. sartorius fibers. Hydrocortisone in conditions of magnesium block of neuro-muscular transmission does not affect the evoked transmission. Hydrocortisone seems to facilitate the evoked quantum acetylcholine secretion only in the presence of curare and in conditions of initial high quantum content of e.p.p. The mechanism of hydrocortisone decurarization effect on neuro-muscular transmission is mainly of a presynaptic nature. PMID- 2987048 TI - The Keyes technique: taking aim at controversy. A DM special interview. PMID- 2987049 TI - Paget's disease of the nipple. Clinical and pathological review of 109 female patients. AB - The most relevant literature concerning Paget's disease of the breast is reviewed. A clinical and pathological study of female patients with Paget's disease of the breast observed during 30 years at the Lisbon 'Instituto Portugues de Oncologia' is carried out and its results are discussed. The low incidence of the disease in this series, when compared with those described in the available literature, is pointed out. The absence of a relation between the type of the cutaneous lesions and the histological type of the underlying tumor is emphasized. The similarity of the microscopical aspects, whichever the histological type of the underlying tumor, is considered. The differences in the clinical behavior of the tumors, related to their histological types, and the differences in respect to age, menarch, menopause, breast feeding, primary lesion and percentage of adenopathies observed between patients with and without a palpable mass, are enumerated. The authors conclude that the cure rate after 5 years is lower in patients with palpable tumors and adenopathies and in the cases with onset before menopause. The cure rate also decreases in relation to the histological type of the underlying tumor. PMID- 2987050 TI - Demonstration of HPV-16 genomes in the nuclei of cervix carcinoma cells. AB - Specimens of 4 different cervical cancers, 1 flat condyloma of the cervix, 1 condyloma acuminatum, 1 morbus Bowen of the skin and 1 skin wart were subjected to in situ hybridization using human papilloma virus type 16 (HPV-16) DNA nick translated with 3H-TTP as a probe. Within each cervical cancer biopsy we found a certain number of tumor cell clusters with clearly labelled nuclei, while sections of the other skin lesions did not reveal any accumulation of grains within their nuclei. Southern blot hybridization of the DNA extracted from the 4 cervical carcinomas with 32p-labelled HPV-16 DNA gave a positive reaction in 3 tumors. Non-reactivity in the 4th biopsy might be due to a low concentration of HPV-genomes within the total extracted DNA of this tissue part. PMID- 2987051 TI - [Eccrine porocarcinoma. Presentation of 4 cases]. AB - Eccrine porocarcinoma is a rare malignant skin tumor arising from the epidermal sweat duct unit (acrosyringium). The report of 4 cases illustrates the clinical and histological characteristics. The prognosis of this carcinoma remains difficult to assess because of the lack of a catamnestic study. PMID- 2987052 TI - Knot security of synthetic absorbable suture material; a comparison of polyglycolic acid and polyglactin-910. AB - To compare the knot reliabilities of two kinds of synthetic absorbable suture material, twelve different knots commonly used in surgical practice were tested. Twenty tests were performed on each knot, i.e., ten with Dexon-S (uncoated) and ten with Vicryl (coated). The knots made with Dexon-S were found to slip less frequently, to be significantly stronger, more consistent, and more reproducible than those made with Vicryl. It was concluded that uncoated Dexon-S is superior to coated Vicryl with respect to know reliability. The practical implications of these findings for surgical performance are discussed. It is emphasized that surgeons should be more aware of the relationship of the holding power of surgical knots to not only the knot-typing technique but also the kind of suture material used. PMID- 2987054 TI - Influence of several perturbants on the rate of autoxidation of horse heart ferrocytochrome c. AB - The effect of several different types of perturbants and pH on the rate of autoxidation of horse heart ferrocytochrome c was investigated. The kinetic behavior is unique to each perturbant used. Rates of autoxidation followed first order kinetics over the time span (0-180 min) studied. The Cl- and Br- anions exhibit an initial increase in the rate of autoxidation up to 100 mM, followed by a decrease in kinetics at 500 mM anion concentration. The ClO4- anion exhibits only an increase in the rate of autoxidation with increasing ionic strength, where as, propylurea, a hydrophobic perturbant, is not effective in altering the rate of autoxidation at equivalent concentrations. These studies suggest that the perturbations of the reduced form of cytochrome c in solution involve mainly non specific solvent effects. PMID- 2987053 TI - NAD+ kinase--a review. AB - NAD+ kinase catalyzes the only (known) biochemical reaction leading to the production of NADP+ from NAD+. Most evidence indicates it is found in the cytoplasm, but reports of its presence in (other) cell bodies can not be discounted. Viewed as a protein, our knowledge of NADK composition and architecture is rudimentary. Though recognized as a large multimeric protein, no agreement is evident for the molecular weight (Mr = approximately 4-65 X 10(4] of the native protein. Is calmodulin an integral subunit of (some, all) NAD+ kinases (analogous to phosphorylase kinase in skeletal muscle)? Or is it an external modulator? Consensus is evident that a subunit of molecular weight 30-35 X 10(3) is a component of the mammalian and yeast kinase. In one case (rabbit liver) two types of subunits are reported to give rise to oligomers differing in molecular weight and catalytic activities. Viewed as an enzyme it is not known why such a complex aggregate is needed for what might otherwise appear to a routine phosphorylation reaction. Rapid equilibrium random (for pigeon liver and C. utilis preparations) and ping-pong (for A. vinelandii kinase) mechanisms have been proposed for the reaction, with multiple reactant binding sites indicated for the random cases. From the perspective of enzyme modulation, the demonstration that green plant and sea urchin egg kinases are targets for calmodulin regulation by intracellular Ca2+ links NADP+ production in these sources to the multi-level discriminatory control functions inherent to this Ca2+ protein complex. Significant questions arise from the results of various investigators considered in this review. These queries offer fertile ground for the selective design of key experiments directed to a better understanding of NAD+ kinase function and pyridine nucleotide biochemistry. PMID- 2987055 TI - Magnitude of ouabain-sensitive respiration of lamb hepatocytes (Ovis aries). AB - In lamb hepatocyte preparations with viabilities greater than 90%, ouabain sensitive respiration accounted for approximately 50% of the total cellular O2 consumption. Lamb hepatocyte preparations with viability of less than 50% exhibited lower (P less than 0.05) total and ouabain-sensitive respiration. The decrease in ouabain-sensitive respiration in these preparations entirely accounted for the drop in total respiration. PMID- 2987056 TI - Significance of glycosidase patterns in lymphoid cells. AB - Characteristic patterns of cell bound lysosomal glycosidases were found in different lymphoblastoid cell lines derived from Epstein-Barr virus-transformed lymphocytes. The enzyme pattern resembled that found in normal lymphocytes from healthy individuals except for a marked increase in alpha-L-fucosidase. beta-D Glucuronidase on the contrary markedly decreased in the lymphoblastoid cells. Burkitt's lymphoma cells on the other hand showed glycosidase patterns that were quite distinct from those in lymphoblastoid cells. Each lymphoma cell line showed a characteristic pattern. This is one indication of a heterogeneous origin of these tumors. Glycosidase patterns may be used to roughly distinguish different lymphoid cell lines. PMID- 2987057 TI - Diurnal variations in the activities of the glycogen metabolizing enzymes in mouse liver. AB - The glycogen level in mouse liver was maximal during the night and decreased to the lowest level during the light period. The peak activity of phosphorylase alpha was observed during the light hours and thus paralleled the decline of hepatic glycogen concentrations. The period of rapid glycogen synthesis (1800 2200 hr) was immediately preceded by maximum glycogen synthase alpha activity. Significant diurnal rhythms for phosphorylase kinase and phosphorylase phosphatase activities were also observed and appear to play a role in regulating the diurnal rhythm of phosphorylase alpha activity. PMID- 2987058 TI - Regulation of processing in multi-functional prohormones: a new hypothesis. PMID- 2987059 TI - Structure and functions of human angiotensin I converting enzyme (kininase II). PMID- 2987060 TI - Lipoproteins and the regulation of steroidogenesis. PMID- 2987061 TI - The role of inositol lipids in hormonal mobilization of cell-associated Ca2+. PMID- 2987063 TI - Inositol lipids and cell proliferation. PMID- 2987062 TI - Phospholipid metabolism in the adrenal cortex. PMID- 2987064 TI - Thyrotropin modulates EGF receptor function in porcine thyroid follicle cells. AB - Porcine thyroid follicle cells in monolayer cultures were shown to contain one single class of high-affinity EGF receptors with Kd = 4.5 X 10(-10) M and approximately 20 000-25 000 receptors per cell. Suspension cultures of aggregated follicle cells, exposed to TSH for 3 days, showed a 3-fold increase in [125I]EGF binding. Scatchard analysis demonstrated that this was due to an increase in receptor number. Other cAMP-elevating agents (cholera toxin, dibutyryl cAMP, forskolin) induced a similar effect. In suspension cultures, preincubation with TSH or cholera toxin for 2 days reduced the subsequent [3H]thymidine incorporation. This inhibition was overcome by a low concentration of EGF (0.1 ng/ml). At higher concentrations of EGF (1-10 ng/ml) the incorporation of [3H]thymidine was potentiated 2-3-fold in cultures preexposed to TSH or cholera toxin. The results demonstrate the presence of a high-affinity EGF receptor in porcine thyroid follicle cells. Receptor expression, as well as responsiveness to the mitogenic action of EGF, is modulated in vitro by TSH, through a cAMP dependent process. PMID- 2987065 TI - Newborn and immature rat testes contain gonadotropin-releasing hormone (GnRH) receptors, and their testosterone production is stimulated by a GnRH agonist in vitro. AB - The possibility of direct gonadal effects of GnRH in the newborn and immature rat testis was studied by two approaches: the presence of specific high-affinity receptors for a GnRH agonist analogue (Buserelin) was studied in animals between the ages of 1 and 60 days; and the effect of Buserelin on testicular testosterone production was studied in vitro. GnRH receptors were found in the testis tissue at all ages studied, and there was a gradual increase in the number of binding sites per testis with advancing age. The highest concentration of GnRH receptors was found in the rat testis tissue on day 1 of life, 3.2 +/- 0.4 fmoles/g tissue (mean +/- SE, n = 5). The binding declined to a nadir, 1.5 +/- 0.5 fmoles/g tissue (in = 10), on day 15 post-partum (p less than 0.05), and thereafter increased again gradually to a level of 2.7 +/- 0.3 fmoles/g (n = 5) on day 60 of life (p less than 0.05). The biphasic concentration curve of GnRH receptors followed closely the volume density of Leydig cells in the developing rat testis. Buserelin at a concentration of 10(-7) M was able to stimulate significantly (p less than 0.01), by 98-146%, the testosterone production of decapsulated testes at all ages studied between 1 and 60 days of life. These results indicate the presence of functional GnRH receptors in the testis tissue of newborn and immature rats. PMID- 2987066 TI - Functional characterization of luteinizing hormone responsiveness and desensitization in perifused interstitial cells. AB - The characteristics of the steroidogenic response of isolated rat testis interstitial cells to repeated 10 min pulses of ovine LH at 2 h intervals were examined in a Bio-gel perifusion system. Maximal responsiveness of the perifused interstitial cells could be maintained for 6-8 h. Thereafter, both basal and LH stimulated testosterone production declined gradually despite supplementation of the perifusion medium with 1% and 5% foetal calf serum or 0.5 microgram/ml insulin. In contrast to the time-related steroidogenic decline, a dose-dependent refractoriness of the interstitial cells could be induced by repeated exposure to LH pulses from 0.01 to 10 ng/ml during the first 6 h of perifusion. The higher the stimulating dose of LH, the greater was the rate and magnitude of the resultant desensitization. With lower doses (0.01 and 0.1 ng/ml) of LH, an initial sensitization or priming effect was also observed. These changes in steroidogenic response occurred in the absence of any significant alterations in LH/hCG receptor binding of the perifused interstitial cells, nor could the refractory state be overcome by stimulation with analogues of cAMP. The perifused interstitial cells, when desensitized with low doses of LH (0.1 ng/ml), were capable of increasing or maintaining testosterone production in response to further stimulation with higher doses of LH (1 and 10 ng/ml). The mechanism(s) underlying the in vitro desensitization of perifused interstitial cells by LH may best be explained on the basis of the interaction between the negative effects of substrate depletion and the positive influence of mobilization of substrate(s) into the metabolically active pool for cholesterol side-chain cleavage. It was concluded that the dose of LH used in the pulsatile stimulation of perifused interstitial cells is critically important not only in determining the total amount of testosterone produced, but also in the pattern of response in terms of the degree of sensitization and desensitization induced. PMID- 2987067 TI - A factor in spent media from Sertoli-cell-enriched cultures that stimulates steroidogenesis in Leydig cells. AB - We investigated whether Sertoli cell spent media (SCM) contain a factor (or factors) which influences steroidogenesis in Leydig cells. Freshly prepared prepubertal interstitial cells or Percoll-purified Leydig cells and similar cells cultured in the presence or absence of LH were incubated for 24 h in the presence of a 5 alpha-reductase inhibitor and in the presence or absence of SCM. The accumulation of C19-steroids (testosterone and androstenedione), C21-steroids (progesterone, 17 alpha-hydroxyprogesterone and 20 alpha-hydroxypregn-4-en-3-one) and cyclic AMP was measured by radioimmunoassay. It could be demonstrated that SCM contains a factor that stimulates an early step in the steroidogenic pathway but at the same time hampers the conversion of C21-precursors into androgens. In freshly prepared Leydig cells the final effect is a stimulation of the androgen output. In Leydig cells cultured in the absence of LH, mainly C21-steroid output is increased. These biological effects resemble those observed with LHRH and its agonists. The activity of the Sertoli cell factor is not affected by an LHRH antagonist, however, and maximally effective concentrations of the factor and LHRH have additive effects, suggesting that they act by distinct receptor systems. Preliminary characterization shows that the factor in SCM is a thermolabile protein with a MW greater than 10 000. The production of the factor decreases during prolonged culture in serum-free medium. Addition of fetal calf serum causes a marked and dose-dependent increase in the production or activity of the factor. Several permanent cell lines (B16, Bowes, BHK, Ratec, RK13, Vero) produce a factor with comparable biological effects on Leydig cells. Nonetheless, the observation that the production of this factor by Sertoli cell cultures is stimulated by FSH and dbcAMP suggests that, in the testis, it may play a role in the paracrine control of Leydig cell function. PMID- 2987068 TI - Preparation of porcine thyroid follicles with preserved polarity: functional and morphological properties in comparison to inside-out follicles. AB - One of the main problems in establishing isolated thyroid follicles in vitro is their tendency to form inside-out follicles. The reason for this change in polarity is unknown. We describe here a method for the preparation of stable thyroid follicles with preserved polarity for at least 6 days. Isolated follicles were obtained by infusion of collagenase (1.5 mg/ml) dissolved in minimal essential medium into the artery of intact thyroid glands. The morphological and functional properties of these follicles were compared to inside-out follicles. These inside-out follicles were obtained by digestion of minced thyroid tissue in a collagenase (1 mg/ml) solution. The polarity of follicles was proved by morphological criteria. Follicles with preserved polarity did not change polarity for at least 6 days in the presence of 1% or 5% fetal calf serum. As the culture conditions for both preparation methods were identical, we conclude that the preparation method rather than the culture condition is responsible for the preservation of cell polarity of isolated thyroid follicles in our system. Increases in cyclic AMP levels induced either by bovine thyrotropin (10(-3) U/ml) or by isoproterenol (10(-6) M) as well as iodide uptake and organification were rapid and significant in right-side-right follicles but not in inside-out follicles. Therefore the TSH receptor and the beta-adrenergic receptor appear to be exclusively located at the basal membrane of follicular cells. In addition, iodide uptake apparently is unidirectional. PMID- 2987069 TI - Clonal variants of differentiated P19 embryonal carcinoma cells exhibit epidermal growth factor receptor kinase activity. AB - Differentiated clonal cell lines were isolated from pluripotent P19 embryonal carcinoma (EC) cells treated as aggregates with retinoic acid. Two were characterized in detail. The lines differ in morphology, proliferation rate, the production of plasminogen activator, and in their mitogenic response to insulin but both produce extracellular matrix proteins and can be serially passaged over extended periods, in contrast to differentiated derivatives of many other EC lines. Further, both lines have receptors for and respond mitogenically to epidermal growth factor (EGF). Endogenous phosphorylation of several proteins, including the EGF receptor (150 kDa) and a 38-kDa protein, is induced by EGF in membranes isolated from these cells. Preincubation of membranes with EGF renders them able to catalyze phosphorylation of tyrosine residues in exogenously added peptide substrates. High voltage electrophoresis confirmed the tyrosine specificity of the phosphorylation on the 150- and 38-kDa bands. By contrast, similar experiments in undifferentiated cells showed that intact P19 EC neither bind nor respond to EGF mitogenically and EGF induces no changes in phosphorylation in isolated membranes. PMID- 2987070 TI - Influence of a transplantable insulinoma on the pancreatic status of insulin and pancreatic polypeptide in the rat. AB - The effect of endogenous hyperinsulinaemia, produced by syngeneic transplantation of rat insulinoma at an extrapancreatic site, on pancreatic insulin and pancreatic polypeptide has been examined by radioimmunoassay and immunohistochemistry. Twenty days after subcutaneous transplantation, tumour bearing rats exhibited marked hyperinsulinaemia and hypoglycaemia, with plasma pancreatic polypeptide concentrations similar to controls. Immunoreactive insulin levels in the head and tail of pancreas of tumour-bearing rats were reduced by 90 95% compared with control animals. Immunoreactive pancreatic polypeptide levels in the head of the pancreas were reduced by 70%, but the relatively low levels of peptide in the pancreatic tail were similar in tumour-bearing and control rats. Insulin and pancreatic polypeptide cells were weakly immunofluorescent in tumour bearing rat pancreas. In conclusion, the presence of an insulinoma at an extrapancreatic site resulted in a severe depletion of endogenous insulin and pancreatic polypeptide, suggesting that there is a functional relationship between the beta and pancreatic polypeptide cell. PMID- 2987072 TI - Hepatic triglyceride lipase and lipoprotein lipase activities in post-heparin plasma of patients with various cancers. AB - The total post-heparin lipolytic activity (PHLA) and hepatic triglyceride lipase (HTGL) and lipoprotein lipase (LPL) activities in post-heparin plasma of patients with various cancers were measured. In patients with cancers, PHLA was similar to that of controls, but the HTGL activity was decreased and the LPL activity was increased. Thus, in cancer patients the ratios of HTGL to PHLA were lower, and the ratios of LPL to PHLA were higher than in controls. No correlation was found between the plasma lipid level and HTGL or LPL activity. PMID- 2987071 TI - Superoxide dismutase and peroxidase are coordinately regulated in differentiated and transformed tissues of Nicotiana tabacum. AB - We used a series of normal and Agrobacterium-transformed, bacteria-free tobacco tissue cultures which differ in their levels of histodifferentiation to test the relationship of superoxide dismutase, peroxidase, and catalase to oncogenic transformation and differentiation. When compared with normal callus, tumor callus contained reduced levels of both superoxide dismutase and peroxidase, and a reduced number of isozymes of both enzymes. Teratomas characterized by limited but abnormal differentiation showed increases in superoxide-dismutase activity and isozymes, but levels of peroxidase activity lower than those found in normal callus despite an increase in the number of peroxidase isozymes. A similar disparity between low peroxidase activity and high isozyme number in the shoot suggests that there are increased levels of peroxidase inhibitors or of molecules which interfere with the spectrophotometric assay for peroxidase in more differentiated tissues. As judged by the number of isozymes of both superoxide dismutase and peroxidase in each tissue, the following conclusions are warranted: first, tobacco copper/zinc superoxide dismutases and peroxidases are encoded in several duplicated loci which are regulated independently. Second, transformation is associated with a decrease in both the specific activity and isozyme number of superoxide dismutase. Third, the partial release from the total inhibition of expression of differentiated function exhibited by teratoma is associated with an increase in both the activity and isozyme number of superoxide dismutase. Finally, the expression of superoxide dismutase and peroxidase isozymes appears to be coordinated during differentiation in a manner that is consistent with their role in an integrated mechanism for the removal of reduced oxygen species. PMID- 2987073 TI - [Papillary adenocarcinoma of Wirsung's duct]. AB - We describe a new case of ductal papillary adenocarcinoma of the pancreas, resected in 1976 (cephalic pancreatoduodenectomy). The patient is still alive. This particular histological type of pancreatic cancer seems to be very rare (9 cases published). The difficulty of preoperative diagnosis and the possibility of long-term postoperative survival clearly differentiate ductal papillary adenocarcinoma from other glandular adenocarcinomas of the pancreas. PMID- 2987074 TI - [Pseudoleukemic hyperleukocytosis disclosing hepatocellular carcinoma]. PMID- 2987075 TI - Extension of hepatoma to the rectus abdominis muscle via ligamentum teres hepatis. AB - The ligamentum teres hepatis connects the umbilicus to the left lobe of the liver, and thus a hepatic lesion can spread through the ligament to the umbilicus and the anterior abdominal wall. We present a case of hepatoma of the left lobe which extended to the rectus abdominis muscle through the ligamentum teres. PMID- 2987076 TI - Localization of small islet-cell tumors. Preoperative and intraoperative ultrasound, computed tomography, arteriography, digital subtraction angiography, and pancreatic venous sampling. AB - A total of 42 islet-cell tumors were examined between 1972 and 1984. Problems of localization were only encountered in 31 tumors less than 2 cm in diameter. Of 31 small tumors, 27 were correctly localized using a combined diagnostic approach: ultrasound was successful in 12/20 tumors, CT in 9/21, angiography in 20/31, intraarterial digital subtraction angiography in 1/2, and pancreatic venous sampling in 13/16. The smallest tumor found by ultrasound and CT was 7 mm in diameter. Intraoperative ultrasound demonstrated all 9 insulinomas examined. Currently, the most useful techniques for localizing small islet-cell tumors are ultrasound, CT, and angiography. CT is particularly useful for tumor staging. Improvement of non-invasive diagnostic techniques will obviate the need for transhepatic blood sampling. PMID- 2987078 TI - [The cytochemistry of granulocytes in acute leukemia]. PMID- 2987077 TI - Gastric carcinoma in Cote d'Or (France). A population-based study. AB - The registry of digestive cancer in the Department of Cote d'Or, France, recorded newly diagnosed cases of gastric cancer between 1976 and 1980. The annual incidence rate, adjusted to the world population, was 15.2/100,000 for men and 6.1/100,000 for women. Stomach cancer incidence showed a substantial decline during the 5 yr of the study and this decline was more pronounced in men than in women. The operability rate was 62.4% and the resectability rate was 40.2%. Operative mortality after curative surgery was 17.6%. For all subjects, 17.6%, 29.1%, and 26.0% had localized, regional, and distant disease, respectively. The remaining 27.3%, not operated upon, with no evidence of metastases, had unclassifiable disease. The overall 5-yr corrected survival rate was 16.5%. In the absence of curative surgery all patients died in the 4 yr after diagnosis. After curative surgery the 5-yr corrected survival rate (excluding operative mortality) was 42.8%. The most important determinant of the survival was the pathological stage of the tumor. The age-corrected 5-yr survival was 98.7% for cases limited to the digestive wall, 45.5% for cases involving the serosa, and 26.6% for cases with locoregional extension. These results support the fact that, although declining, gastric cancer remains relatively frequent. Its overall prognosis in a well-defined population, where cases limited to the digestive wall are rare, remains poor. PMID- 2987079 TI - The effects of lidocaine and procainamide on the sarcolemmal membrane high affinity cyclic AMP phosphodiesterase of rat myocardium. AB - The incubation of rat myocardial sarcolemmal membranes with 10(-4)M lidocaine or procainamide results in a decreased fluorescence polarization of the extrinsic probes (diphenylhexatriene and 12-anthroylstearate) and in the stimulation of the membrane-bound high affinity cyclic AMP phosphodiesterase activity (PDE). Lidocaine or procainamide do not contrast, but facilitate the aminophylline inhibitory action on the PDE activity, without modifying the basic molecular mechanism of the inhibitory action. The amplitudes of the effects described are inversely correlated to the temperature, being greater at the lower tested temperature (25 degrees greater than 30 degrees greater than 37 degrees C). PMID- 2987080 TI - Prostaglandin A2 receptor in rat intestinal basolateral plasma membranes. AB - The specific prostaglandin A2 receptor was presented in rat intestinal basolateral plasma membrane. The dissociation constant was 4.39 X 10(-8) M and the value of the binding capacity was 3.33 pmol/mg protein. [3H]prostaglandin A2 binding was specifically inhibited by Na+ or ATP. The inhibition manners of Na+ and ATP for [3H]prostaglandin A2 binding were of a competitive and noncompetitive type, respectively. PMID- 2987081 TI - Endogenous opioid nomenclature: light at the end of the tunnel. PMID- 2987082 TI - Ultrastructural demonstration of iodine binding and peroxidase activity in the endostyle of Oikopleura dioica (Appendicularia). AB - The endostyles of cephalochordates, ascidians, and larval petromyzontids have the capacity to organify iodine. A similar mechanism in the appendicularian endostyle has hitherto been unknown. Observations in this study of Oikopleura dioica with electron microscopic autoradiography and cytochemistry show that also the appendicularian endostyle has iodinating capacity and that the iodinating cells contain peroxidase, an enzyme responsible for iodination. After incubation in seawater containing 125I-, autoradiography revealed a selective labeling in the dorsal portion of the endostyle. The endostyle of O. dioica is on each side lined by four rows of corridor cells. The autoradiographic grains were mainly located over the endostylar lumen or associated with the luminal surface of the two central rows of corridor cells. These cells, but no other endostylar cells, also showed a positive reaction for peroxidase. The reaction product was distributed along the luminal plasma membrane and was also present in the cytoplasm within rough endoplasmatic reticulum, Golgi apparatus, and vesicles. The selective labeling as well as the cytochemical reaction were abolished by incubation in methimazole, an inhibitor of peroxidase. It is suggested that the two central rows of the corridor cells can be considered as homologs to iodine-binding zones in other endostyles and also as a primitive forerunner to the vertebrate thyroid gland. PMID- 2987083 TI - Effects of N-terminal peptide of salmon proopiocortin on interrenal function of the rainbow trout. AB - Effects of N-terminal peptide of salmon proopiocortin (salmon NPP-I) on cortisol secretion was examined in vitro using diced interrenal tissue from the rainbow trout, Salmo gairdneri. ACTH(1-24) at concentrations of 1 to 50 nM stimulated cortisol secretion in dose-dependent manner, whereas salmon NPP-I had no effect over a range of 50 pM to 500 nM. Cortisol secretion in response to various doses of ACTH(1-24) was modified slightly when 1 to 100 nM of salmon NPP-I was added to the incubation medium together with ACTH. An augmentation of in vitro secretion of cortisol in response to ACTH(1-24) was observed when the interrenal was removed from the trout pretreated with one IU of porcine ACTH but not with 10 micrograms of salmon NPP-I. A slight but significant potentiating effect of salmon NPP-I (10 or 100 nM) on the ACTH-induced cortisol secretion was observed when the trout was sensitized to ACTH by porcine ACTH pretreatment. Furthermore, six daily injections of salmon NPP-I into the trout induced hyperplasia of interrenal tissue. These findings suggest that NPP-I, together with ACTH, may be involved in controlling interrenal function in the trout. Such activities could be due to conservative region in the N-terminal portion of NPP. PMID- 2987084 TI - Genetic differentiation of transposable elements under mutation and unbiased gene conversion. AB - A model is developed to predict the extent of genetic differentiation in a family of transposable elements under the combined effects of genetic drift, transposition, mutation and unbiased gene conversion. The model is based on simplifying assumptions that are valid when transposition is always to new sites and copy number per site is low. In the absence of gene conversion, the degree of differentiation as measured by the probability of identity of different elements is the same as at a single locus with the same mutation rate but in a population of effective size Nc/2, where N is the population size and c is the number of copies per individual. The inclusion of unbiased gene conversion does not significantly change this result. If, as seems to be the case, families of transposable elements are relatively homogeneous, then the model implies either that mutation rates for transposable elements are much lower than at comparable single-copy loci or that some other force, such as natural selection or biased gene conversion, is at work. Transposition is a very ineffective force for homogenizing a family of transposable elements. PMID- 2987085 TI - Molecular cloning of the delta-endotoxin gene of Bacillus thuringiensis var. israelensis. AB - A transformant of Bacillus megaterium, VB131, was isolated which carries a 6.3-kb XbaI segment of the crystal toxin gene of Bacillus thuringiensis var. israelensis (BTI) cloned in a vector plasmid pBC16 to yield pVB131. The chimeric plasmid DNA from VB131 was introduced into a transformable Bacillus subtilis strain by competence transformation. Both the B. megaterium VB131 strain and the B. subtilis strain harboring the chimeric plasmid produced irregular, parasporal, phase-refractile, crystalline inclusions (Cry+) during sporulation. The sporulated cells as well as the isolated crystal inclusions of the pVB131 containing B. megaterium and B. subtilis strains were highly toxic to the larvae of Aedes aegypti. Also, the solubilized crystal protein preparation from VB131[pVB131] showed clear immuno cross-reaction with antiserum to the BTI crystal toxin. 32P-labeled pVB131 plasmid DNA showed specific hybridization with a 112-kb plasmid DNA of Cry+ strains of BTI, and no hybridization with other plasmid or chromosomal DNA of either Cry+ or Cry- variants. These results are in agreement with our previous findings (Gonzalez and Carlton, 1984) that the 112-kb plasmid of BTI is associated with the production of the crystal toxin. PMID- 2987086 TI - A study of constipation in the elderly living at home. AB - A cross-sectional survey of bowel habit has been carried out on 201 elderly patients living at home. Although symptoms of constipation were common, reported bowel frequency was similar to younger people. Constipation was most clearly associated with poor mobility and depression. Digital rectal examination was generally unreliable as an indicator of constipation. Using the abdominal radiograph as the final arbiter, true constipation was present in less than half of those complaining of constipation. PMID- 2987087 TI - Beta-adrenergic receptor subtypes iris-ciliary body of rabbits. PMID- 2987088 TI - Isolation and characterization of the pMI10 plasmid from Bacillus subtilis, an alpha-amylase producer. AB - The plasmid DNA pMI10 (5310 bp) was isolated from the alpha-amylase producing strain B. subtilis A18. Thirteen restriction endonucleases were used to digest pMI10 DNA and the restriction map of pMI10 DNA was constructed by mapping PstI (1), HindII (2), BglI (2), BspRI (3) and HindIII (3) sites. PMID- 2987089 TI - [Peculiarities of mamillary secretion and cytodiagnosis]. PMID- 2987090 TI - [Effects of anti-tumor drugs and gastrointestinal hormones on the growth of pancreatic duct cell adenocarcinoma in homologous transplanted animal models]. AB - The pancreatic duct cell adenocarcinoma induced by di-isopropanol nitrosamine in Syrian golden hamsters could be easily and repeatedly into the subcutaneous tissues or the pancreas of the homologous animals. We examined the anti-tumor effects of the administration of FT-207 and the coadministration of FT-207 and uracil on the pancreatic cancer transplanted into the subcutaneous tissues and the pancreas. The inhibitory effect on the tumor growth observed in the group treated with FT-207 and uracil was more striking than that in the group treated with only FT-207. Such enhanced anti-tumor effect observed in the coadministration group seemed to be due to the increased 5-FU concentration in the tumor tissues. Some gastrointestinal hormones (cholecystokinin, caerulein, secretin and tetragastrin) injected intraperitoneally promoted the growth of subcutaneously transplanted tumor. These gastrointestinal hormones except tetragastrin also increased the weight of the pancreas. In an analysis of RNA and DNA contents and RNA/DNA ratio in tumor tissues, the promotion of the tumor growth seemed to stem from hyperplasia and/or hypertrophy of the cancer cells. Therefore, it is suggested that some gastrointestinal hormones have trophic actions on the pancreatic duct cell adenocarcinoma as well as normal pancreas. PMID- 2987091 TI - [Studies for clinical application of ovine corticotropin-releasing factor]. AB - Synthetic ovine corticotropin-releasing factor (o-CRF) stimulated adrenocorticotropin (ACTH) release at the concentration of 10(-10) M or more in monolayer culture of rat anterior pituitary cells. Dexamethasone, 10(-7) M, inhibited this effect. In 5 healthy human subjects, o-CRF, 1 microgram/kg iv bolus, increased plasma ACTH levels from less than 10 pg/ml to 40.4 +/- 11.0 (mean +/- SD) after 30-60 min, and plasma cortisol from 12.8 +/- 2.8 micrograms/dl to 23.6 +/- 3.1 after 45-60 min. Of 7 patients with Cushing's disease (CD), five showed an exaggerated response of plasma ACTH and cortisol, one an exaggerated response of plasma ACTH but low response of plasma cortisol and the other no response of both hormones. The significant positive correlation between the inhibition of plasma cortisol by dexamethasone and the response of plasma ACTH and cortisol to o-CRF in CD was seen. No response of plasma ACTH and cortisol to o-CRF was seen in each one patient with Cushing's syndrome due to an adrenocortical adenoma, ectopic ACTH syndrome (but low response at retesting), isolated ACTH deficiency and Sheehan's syndrome. In one patient with Addison's disease an exaggerated response of plasma ACTH but no response of plasma cortisol was seen. In 4 of 5 healthy subjects and 5 of 7 patients with CD, plasma ACTH and cortisol levels showed a second peak at 120--210 min after o-CRF administration. To clarify the prolonged effect of o-CRF in human in vivo, the disappearance rates of injected o-CRF were evaluated by radioimmunoassay in 3 patients with cured Cushing's syndrome. A biexponential decay curve showed t1/2 values of 9.3 +/- 0.4 min and 79.6 +/- 0.7 min (mean +/- SE). From the chromatographic profile, a portion of injected o-CRF was thought to be bound to macromolecule (s). o-CRF, as a specific secretagogue of ACTH, is thought to be useful tool in evaluating patients with hypothalamo-pituitary-adrenal disorders. PMID- 2987092 TI - Concentration of native prolactin and prolactin binding sites in hepatic subcellular fractions from hyperprolactinemic rats. AB - Lactogen binding and prolactin content were measured in hepatic subcellular fractions from tumor-bearing rats (TBR; MtT/F4, MtT/W5, MtT/W10) with elevated prolactin and growth hormone levels and from control animals. Specific binding of 125I-oPRL to Golgi fractions from tumor-bearing animals was 2.5 to 7 fold greater than that from controls. Binding to plasmalemma was 6-fold greater in tumor bearing rats. The specific binding of 125I-labelled bGH and insulin showed less marked differences between TBR and controls. Subcellular fractions were extracted with HCl to determine hormonal content. The content of prolactin and growth hormone in Golgi fractions from TBR was at least 20-fold that in fractions from controls. Rat prolactin extracted from Golgi heavy elements was 50% as effective as native material in binding to lactogen receptors as judged by radioreceptor assay. These studies demonstrate that the chronic elevation of prolactin was associated with an increase of receptors not only in the intracellular compartment but on the cell surface as well. Furthermore, they demonstrate that native prolactin is internalized and accumulated in rat liver Golgi fractions. PMID- 2987093 TI - Hormone-dependent transformation and nuclear localisation of 1,25 dihydroxyvitamin D3 receptors from human breast cancer cell lines and chick duodenum. AB - Recent studies on the 1,25-dihydroxyvitamin D3 (calcitriol) receptor have shown association of unoccupied receptor with isolated nuclei, thus suggesting that hormone is not required for transformation and nuclear localisation of this receptor. In the present work calcitriol receptors from cultured breast cancer cells were studied for evidence of hormone-dependent activation and compared to those from chick duodenum. Unlike other steroid receptors changes in receptor mobility on ion exchange and gel filtration were not found for occupied and unoccupied receptors. Furthermore no changes in affinity were observed on DNA cellulose with both hormone-bound and unoccupied receptor having equally high affinity, eluting at 0.25 M KCI. However, a substantial hormone-dependent increase in receptor affinity for nuclei was seen. Thus calcitriol receptors do appear to undergo hormone-dependent transformation which is detected by their increased affinity for nuclei, without any accompanying gross changes in charge density, size or affinity for DNA-cellulose. Previously, we have reported that fractionation of T-47D cells in a low salt buffer resulted in recovery of unoccupied receptors in the cytosol, whereas occupied receptors were associated with purified nuclei. The data presented in this paper and our previous work suggest that calcitriol receptors do undergo a hormone-dependent increase in their affinity for nuclei. Furthermore in all this work calcitriol receptors from cultured human breast cancer cells displayed identical physicochemical characteristics to those of chick duodenal receptors. PMID- 2987094 TI - Hormone and haemodynamic effects of angiotensin II infusion during captopril treatment for heart failure. AB - Withdrawal of captopril therapy for cardiac failure results in increments in plasma cortisol, noradrenaline and heart rate. To determine whether these changes related to the concomitant rise in circulating angiotensin II, we infused angiotensin II at 0.5, 2, 4 and 8 ng/kg/minute, each infusion lasting for 1 hour, in 4 patients during maintenance captopril therapy for heart failure. A control solution of 5% dextrose was infused over a similar time interval on a separate day. The study was performed under metabolic balance conditions, with constant body posture and continuous haemodynamic monitoring. Angiotensin II induced the expected rise in arterial pressure and in plasma aldosterone. In contrast the diurnal decline in plasma ACTH and cortisol was not altered, and no changes in noradrenaline or heart rate were observed. Plasma angiotensin II appears to have little or no effect on ACTH, cortisol, noradrenaline and heart rate under the conditions of this study. PMID- 2987095 TI - Polyamines stimulate hGH receptor-binding in liver microsomes. PMID- 2987096 TI - Pharmacokinetics and biotransformation of estradiol valerate in ovariectomized women. AB - Estradiol valerate is well suited for treatment of the characteristic symptoms accompanying menopause in women. The pharmacokinetics and biotransformation of estradiol valerate were studied in women following intravenous, intramuscular and oral administration. Intravenously given, estradiol valerate is split by enzymatic hydrolysis into 17 beta-estradiol and the fatty acid. The estrogen arising in vivo from the steroid ester is subject to intermediate metabolism. The metabolites are eliminated at a nearly constant rate mainly in urine. The biotransformation of estradiol valerate was not different following intravenous or intramuscular injection. Orally given estradiol valerate is subject to an extensive first pass metabolism. Daily repeated oral administration does not result in accumulation of 17 beta-estradiol and its metabolites. PMID- 2987097 TI - Stimulatory action of follicular fluid components on maturation of granulosa cells from small porcine follicles. AB - Follicular fluid obtained from large (6-12 mm) porcine follicles (LFF) was investigated to determine its stimulatory activity on progesterone secretion and on follicle stimulating hormone (FSH) induction of 125I-human chorionic gonadotropin (hCG)-luteinizing hormone (LH) binding sites in porcine granulosa cells in a 4-day culture. Incubation of granulosa cells harvested from small porcine follicles (1-2 mm) with 50% LFF led to stimulation of LH/hCG binding sites and progesterone secretion. After partial purification of pooled LFF or proteins precipitated with 90% ethanol on Sephadex G-100 the greatest stimulatory activity was found in the second protein peak eluted from the column. Chromatography of part of the active fraction on DEAE Sephacel using a continuous gradient of NH4HCO3 yielded seven protein fractions. The second fraction, which eluted early, contained the majority of the stimulatory activity which was purified about 32-fold compared to native LFF. In contrast, addition of follicular fluid recovered from small porcine follicles inhibited FSH induction of LH/hCG binding sites and progesterone secretion. It can be concluded, that maturation of granulosa cells from small follicles may be enhanced by protein(s) present in LFF, but not in fluid recovered from less mature follicles. PMID- 2987098 TI - Human hepatoma-associated cell surface antigen: identification and characterization by means of monoclonal antibodies. AB - A library of murine monoclonal antibodies reactive with human hepatoma cells was generated following immunization of Balb/c mice with an intact cloned human hepatoma cell line, designated PLC/PRF/5-NR. We report the characterization of one such IgG2a antibody, designated anti-PLC1. This antibody specifically stains parental PLC/PRF/5 cell membranes and membranes of SK-Hep 1 and Mahlavu human hepatoma cells grown in culture, using indirect immunofluorescence and horseradish immunoperoxidase techniques. A similar pattern of membranous staining was observed in solid tumors derived from the three hepatoma cell lines which were injected subcutaneously into athymic nude rats and mice. Spontaneous capping on the cell surface was observed in 7 to 30% of the three human hepatocellular carcinoma cell types when incubated in suspension with monoclonal anti-PLC1 at 37 degrees C. Treatment of cells with trypsin or sustained growth in culture did not affect the intensity of membranous staining. Monoclonal anti-PLC1 appeared specific, and antibodies did not stain a variety of human carcinoma cell lines and primary tumors of nonhepatic origin, or several normal human and murine tissues. Purified 125I-labeled monoclonal anti-PLC1 bound specifically to the three hepatoma cell lines in culture. Specificity of the antigen-antibody reaction was demonstrated by competitive binding inhibition in experiments using unlabeled homologous antibody. Binding of 125I-anti-PLC1 was not inhibited by unlabeled monoclonal antibodies to HBsAg or to alpha-fetoprotein. Two hepatoma cell lines secrete a protein that specifically blocks binding of 125I-anti-PLC1 antibodies to cell surface antigenic determinants. This "hepatoma-associated" protein was subsequently purified by affinity chromatography from supernates derived from the three hepatoma cell lines.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987100 TI - Tubular carcinoma of the breast. A long term follow-up. AB - The aim of this study is to provide a long-term follow-up of patients with tubular carcinoma and to investigate whether the separation of the mixed type of tubular carcinoma, tubular component more than 75%, from invasive ductal carcinoma with a major tubular component (ductal carcinoma MTC), tubular component 50 to 75%, is supported by long term follow-up. For this study 388 consecutive breast carcinomas were selected from the files of the Department of Pathology, Bispebjerg Hospital, Denmark for the years: 1956-59. Twenty tubular carcinomas were found and 16 ductal carcinomas MTC were also identified. For each patient with tubular carcinoma and ductal carcinoma MTC, two controls with ductal carcinoma NOS were selected. The patients in these four groups were followed until 1 April 1982. None of the patients were lost to follow-up. Among the 20 patients with tubular carcinoma, 10 died within this period compared to 37 of the 40 controls. Of the 16 patients with ductal carcinoma MTC, 15 died compared to 29 of the 32 controls. The difference in Kaplan-Meier survival curves between the tubular carcinomas and their controls is highly significant. There is no significant difference between ductal carcinomas MTC and their controls. These findings are not changed significantly when 5, 10, and 20 year survival rates are corrected for expected survival. This study demonstrates long-term survival for patients with tubular carcinoma as compared to ductal carcinoma NOS. There is no difference in long-term survival between ductal carcinoma MTC, tubular component between 50 and 75%, and ductal carcinoma NOS. It therefore seems appropriate to set 75% tubular component as a reasonable cut-off point for the mixed type of tubular carcinoma. PMID- 2987099 TI - Hepatic (Na+,K+)-ATPase: a current view of its structure, function and localization in rat liver as revealed by studies with monoclonal antibodies. PMID- 2987101 TI - Elderly require shift in hospital resources. PMID- 2987102 TI - Lung cancer heterogeneity: a blinded and randomized study of 100 consecutive cases. AB - The heterogeneity of lung carcinomas was recognized in the past, but few previous studies attempted to quantitate this heterogeneity. In the present study 100 consecutive cases of lung carcinoma (65 surgical resections and 35 autopsies) were collected, and either the entire tumor or ten blocks were examined in a blinded and randomized fashion using the revised (1981) WHO classification. At least three of five panelists agreed on the major histologic type present for 94 per cent of the slides. Agreement for the diagnosis of small cell carcinomas (at least four of five observers) was 98 per cent, but only 72 per cent agreement was attained for the subtyping of small cell carcinomas (e.g., oat cell versus intermediate). Only 34 per cent of the cases were homogeneous according to the majority of the panelists. An additional 21 per cent of the cases showed minor (subtype) heterogeneity (e.g., mixtures of acinar and papillary patterns in adenocarcinoma). Forty-five per cent of the cases showed major heterogeneity, i.e., at least one slide from the case showed a major histologic type different from that of the remainder. Seven small cell carcinomas were homogeneous, whereas in eight cases mixtures of small cell and other cell types were seen. In all but one of the cases involving bronchioloalveolar cell patterns, other patterns of adenocarcinoma were present elsewhere in the tumor. In all six cases involving giant cell carcinoma patterns, adenocarcinoma patterns were also present in some sections. Heterogeneity was identified by extensive sampling of the entire tumor and was seldom recognized in biopsy specimens. PMID- 2987103 TI - Metastasizing placental site trophoblastic tumor: a case study. AB - The clinicopathologic features of placental site trophoblastic tumor (PSTT), a rare form of trophoblastic disease, were systematically described only relatively recently. Previously, in endometrial curettage and hysterectomy specimens, PSTT was not distinguished from choriocarcinoma or sarcoma, although in most cases the natural history had been benign. A case of biopsy-proved metastasis to para aortic lymph nodes with radiologic evidence of lung metastasis is reported. Histologically, the tumor appeared similar to the tumors in other reported cases of PSTT. The mitotic rate was variable, being very high in a few small fragments obtained by curettage. Flow cytometric analysis of the tumor showed cells with diploid DNA content and a relatively low proliferative fraction. The tumor maintained its morphologic and functional characteristics following xenotransplantation into Balb/c nu/nu mice. In spite of a treatment regimen that included hysterectomy and multiagent chemotherapy, the patient continued to have evidence of persistent trophoblastic disease 15 months after presentation. PMID- 2987104 TI - Two new polymorphic markers in the human pro alpha 2(1) collagen gene. AB - Structural defects in the human type 1 collagen genes are known to be the cause of several inherited disorders of connective tissue, such as osteogenesis imperfecta. The analysis and prenatal diagnosis of these disorders would be facilitated by establishing a set of polymorphic markers at these gene loci. We have previously reported the presence of an Msp 1 restriction fragment length polymorphism in the pro alpha 2 (1) collagen genes of several Southern African populations (Grobler-Rabie et al., in press). This report describes the detection of a Bgl II and an EcoRI polymorphism in the pro alpha 2 gene of South African Blacks. PMID- 2987105 TI - X-linked dominant Charcot-Marie-Tooth disease: suggestion of linkage with a cloned DNA sequence from the proximal Xq. AB - A large kindred with the X-linked dominant form of peroneal muscular atrophy (Charcot-Marie-Tooth disease) was analyzed for individual variation in the length of DNA fragments after restriction endonuclease digestion. A systematic search was performed for linkage with a series of cloned single-copy DNA sequences of known regional assignment to the human X chromosome. Close linkage was found with the pDP34 probe (DXYS1 locus, Xq13-q21), suggesting that the gene responsible for the disease is located on the proximal long arm of the X chromosome. PMID- 2987106 TI - A new restriction fragment length polymorphism in the haptoglobin gene region. AB - Direct gene analysis of the haptoglobin gene region was carried out by Southern blotting using an Hp cDNA as probe. Two types of polymorphism were observed: one due to intragenic duplication, is characterized by a constant fragment length difference of 1700bp observed with several enzymes and by complete correspondence with the protein molecular weight polymorphism; the second type, due to point mutation, was represented by two additional restriction sites for Eco RI and Pst I, with a frequency comparable to that of other genes. These two mutations segregated together in families, suggesting that the recently described Hp related gene is closely linked to the Hp gene. Moreover, they were completely associated with each other. The evolutionary significance of this finding is discussed. PMID- 2987107 TI - Evidence for a "new" allele at the phosphoglycolate phosphatase locus. AB - An obviously new phosphoglycolate phosphatase (PGP) gene product (PGP Sumatra) was detected by use of horizontal starch gel electrophoresis (SGE). The observed phenotype PGP (1-Sumatra) can be distinguished from any known PGP type. PMID- 2987109 TI - Anti-hormone anti-idiotypes are probes for receptor structure and function. AB - A class of anti-idiotypes raised against antibodies specific for hormones or drugs would be anticipated to interact with their respective receptors. We have raised anti-TSH anti-idiotypic antibodies and found these to interact with the high affinity binding site for TSH on thyroid membranes, to induce adenylate cyclase activation and iodide transport into dispersed thyroid cells as well as to promote their organization into follicular structures. The anti-TSH anti idiotypic antibody interacted with an Mr approximately 200,000 holoreceptor on protein blots of thyroid membranes resolved on SDS-PAGE in the absence of reductant. In another set of experiments we raised anti-idiotypic antibodies against monoclonal antibodies specific for the alpha and beta subunits of TSH, respectively. Neither the alpha nor beta monoclonal antibody-specific anti idiotypic antibodies interacted with the TSH holoreceptor. The combinations of the two anti-idiotypic antibodies, however, did so and increased basal cyclase activity compared to normal IgG and individual anti-idiotypes as well as mediating TSH-specific cAMP dependent physiologic changes. As a result of the second set of experiments, we propose that the interaction of TSH with its receptor involves two cooperative signals delivered by the two subunits rather than a single signal requiring their combination. Anti-idiotypic antibodies raised against highly purified hormones can be obtained in large amounts. They facilitate simple isolation of hormone receptors and are useful as probes for hormone-receptor interactions. PMID- 2987108 TI - Regulatory roles of human OKT4/OKT8 subsets in polyclonal immunoglobulin production induced by herpes simplex type 1 virus. AB - T cells, OKT4 cells and OKT8 cells from the peripheral blood of normal individuals seropositive for herpes simplex type 1 virus (HSV) were studied for their capacity to regulate in vitro polyclonal immunoglobulin (Ig) production induced by inactivated HSV. Polyclonal Ig production induced by HSV has been demonstrated to be T-cell dependent. T cells, OKT4 cells and OKT8 cells were co cultured with autologous non-T cells in the presence of HSV or pokeweed mitogen (PWM) and the number of plaque-forming cells (PFC) was measured with an hemolytic plaque assay after 6 days of culture. The results in the HSV system show that the OKT4 cells provided significantly more helper activity than OKT8 cells (p = 0.002); and the OKT8 cells exhibited more suppressor activity than OKT4 cells for Ig production (p = 0.02). The helper activity of OKT4 cells after HSV stimulation was significantly less than that obtained after pokeweed mitogen stimulation (p = 0.01). The in vitro polyclonal immunoglobulin response to HSV antigen is regulated by the balance of helper/suppressor activity exerted by OKT4 and OKT8 cell subsets. PMID- 2987110 TI - Lack of effect of LTB4 on human natural killer cell activity and T lymphocyte transformation. AB - Human natural killer (NK) cell activity was not significantly affected by leukotriene (LT) B4 over a wide concentration range (10(-6)-10(-14) M), whether added directly to the NK cell assay or after preincubation of effector cells for 2 h with the drug before addition of Cr labelled K562 target cells. In addition, LTB4 did not affect the kinetics of NK cell mediated cytotoxicity. Addition of LTB4 (10(-6)-10(-10) M) to concanavalin A (Con A) or phytohaemagglutinin (PHA) stimulated human peripheral blood mononuclear cells (PBMC) had no significant effect on proliferation measured by tritiated thymidine incorporation, using both optimal and sub-optimal mitogen concentrations. Whilst it is clear that LTB4 is an important mediator of inflammation involving polymorphonuclear (PMN) leukocytes in vivo, it does not affect NK cell or T cell function in vitro. PMID- 2987112 TI - The effect of several uremic parameters on uremic serum inhibition of lymphocyte blastogenesis. AB - Uremic sera are known to inhibit thymidine incorporation of normal lymphocytes. The nature of the factor(s) responsible for this inhibitory effect has not been completely elucidated. In this study a possible correlation was investigated between a number of uremic blood constituents altered with the progression of the disease and the immunoinhibitory effect of the respective sera. No such correlation was found with the values of hematocrit, urea, creatinine, calcium and phosphorus. On the other hand a significant negative correlation emerged between H+ and Mg2+ ion levels and the inhibition imposed on normal lymphocyte thymidine incorporation. This apparently paradoxical result would indicate that with regard to these two parameters the greater the severity of renal failure the smaller would be the immunoinhibitory effect of the respective serum. The inhibition imposed by uremic serum on immune functions is probably a multifactorial phenomenon, in which H+ and Mg2+ might play a role antagonistic to inhibitory factors. PMID- 2987111 TI - Leukocyte migration inhibitory factor production by activated lymphocytes representing immunological memory or virus-receptor interaction: response of T cell subsets to Epstein-Barr virus nuclear antigen, response of B cells to UV inactivated Epstein-Barr virus. AB - Both T and B lymphocytes are known to produce leukocyte migration inhibitory factor (LIF) after appropriate activation. We showed that EBV nuclear antigen (EBNA) triggered T cells for LIF production in an immunologically specific way: only T cells of seropositive individuals responded. Both Fc receptor positive and negative T cells produced LIF, and the presence of macrophages was necessary. The virus itself activated B cells independently of the serological status of the donors, thus the function was not based on immunological memory. This phenomenon was independent of the transforming capacity of the virus, because UV-inactivated virus also elicited LIF production by B lymphocytes. This triggering seems to be the consequence of the virus-receptor interaction on the cell surface. PMID- 2987114 TI - Genetic polymorphism at the kappa chain locus in mice: comparisons of restriction enzyme hybridization fragments of variable and constant region genes. AB - Variable (V kappa) and constant (C kappa) region genes of the mouse kappa light chain have been compared in inbred strains and in geographically isolated or genetically separated populations of mice by Southern blot analysis of endonuclease-restricted germline DNA. In most cases, the C kappa gene is found on a single restriction fragment while the V kappa genes of the V kappa 19 and V kappa 21 groups are each found on several (6-18) fragments. The restriction fragment (RF) patterns of V kappa 19 and V kappa 21 groups are both polymorphic when compared among inbred mouse strains. Southern blot patterns of V kappa 21 and V kappa 19 of inbred strains are also found among some geographically isolated populations of mice, suggesting that inbred strains acquired kappa loci from different subspecies. Some populations of geographical isolates show V kappa 21, V kappa 19, and C kappa contexts similar to inbred mice while more distantly related species within the genus Mus and laboratory rats show no apparent similarity in context to inbred strains. Variable region genes determining the RF patterns of V kappa 19 and V kappa 21 appear to be linked to each other and to the C kappa and Lyt-3 loci. PMID- 2987116 TI - T-cell-fibroblast hybridoma deformability and concanavalin A-induced agglutination. AB - Cell adhesion influences many important immunological functions such as phagocytosis or T-cell-mediated cytotoxicity. Previous work suggested that the ease of inducing intercellular bonds (i.e. binding efficiency) and the difficulty to separate bound cells with mechanical forces (i.e. binding strength) might be parameters of different significances. The present report describes a study made on two T-lymphocyte/polyoma virus transformed fibroblast hybrid subclones (3D1c and 3D1n) with markedly different adhesive properties: indeed, 3D1c cells were at the same time more readily agglutinated with concanavalin A and more easily disagglutinated than 3D1n. In order to understand these differences, a systematic comparison of various properties of 3D1c and 3D1n cells was undertaken. The following parameters were studied: surface density of concanavalin A binding sites, surface electrostatic charge, hydrophobicity, fluorescence polarization measured on individual cells, and ability to spread on a flat substrate in response to volume or surface forces. It is concluded that cell deformability and/or spreading ability might be an important determinant of binding strength, but the factors governing binding efficiency remained incompletely understood. It is suggested that the methods described in the present report might help understanding differences between various tumor cell lines with different malignant potential. PMID- 2987113 TI - Subline divergence within L.C. Strong's C3H and CBA inbred mouse strains. A review. PMID- 2987115 TI - The HLA-AW24 gene: sequence, surroundings and comparison with the HLA-A2 and HLA A3 genes. AB - A cosmid clone containing two class I sequences was found to cause expression of the HLA-AW24 protein after transfection into mouse L cells. The restriction map of this cosmid shows extensive homology over 26 kb with the map of the HLA-A3 region obtained from cosmids of the same library, constructed with DNA from an HLA-A3/HLA-AW24 heterozygote, but diverges over the remaining 14 kb. The HLA-AW24 gene was subcloned from this cosmid and its nucleotide sequence was determined. Amino acid and, more strikingly, nucleotide sequence comparisons with other HLA alleles indicate that the A locus alleles are more closely related to each other than to alleles from other HLA loci. A very skewed distribution of silent substitutions is apparent, and the occurrence of clustered multiple substitutions hints at gene-conversion-like events. PMID- 2987117 TI - Serial serum 5'-nucleotidase and S.G.O.T. levels in acute myocardial infarction. PMID- 2987118 TI - alpha 1-Adrenergic receptor binding in the spontaneously hypertensive rat. AB - Increased sympathetic outflow from the central nervous system to the periphery may contribute to the initiation of hypertension in spontaneously hypertensive rats (SHR). As this alteration in sympathetic activity may be mediated in part by alpha-adrenergic receptors in the central nervous system, the current study examined alpha 1-adrenergic receptors in various brain areas of SHR and normotensive Wistar-Kyoto control rats (WKY). The alpha 1-adrenergic receptor number and apparent affinity constants of brain sections of both young prehypertensive animals (4 weeks old) and mature hypertensive animals (12 weeks old) were studied with the alpha 1-adrenergic receptor antagonist [3H]WB-4101 to label the alpha-adrenergic receptor. Five brain regions were studied: rostral hypothalamus, caudal hypothalamus, locus ceruleus, nucleus tractus solitarius, and frontal cortical poles. In comparison to normotensive controls, mature hypertensive rats had a significantly greater density (p less than 0.05) of the alpha 1-adrenergic receptors in the rostral hypothalamus (+11%), caudal hypothalamus (+25%), and frontal cortical poles (+20%). Significantly greater (p less than 0.05) alpha 1-adrenergic receptor density was found in the rostral hypothalamus (+27%), caudal hypothalamus (+60%), and locus ceruleus (+39%) of the young prehypertensive SHR compared with age-matched WKY. These results indicate the presence of altered adrenergic receptor systems in the brains of genetically hypertensive animals and suggest that changes in the receptor systems take place during establishment of the hypertension. PMID- 2987119 TI - Renal and blood pressure responses to synthetic atrial natriuretic factor in spontaneously hypertensive rats. AB - Atrial natriuretic factor (ANF) is a potent natriuretic and vasorelaxant agent that also stimulates guanosine 3',5'-cyclic monophosphate (cGMP) excretion in normotensive animals. These properties suggest that ANF may be involved in the regulation of blood pressure. To test a pure preparation of ANF in both normotensive and hypertensive animals, a synthetic 26 amino acid peptide (sANF) contained within endogenous rat ANF was infused intravenously into conscious Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) at doses from 12 to 190 pmol/minute. Mean arterial pressure fell progressively as doses of sANF were increased until maximum responses of -41 +/- 5 mm Hg and -29 +/- 5 mm Hg were obtained during infusion of 95 pmol/minute sANF in SHR and WKY, respectively. Heart rate was not significantly affected in either group. At sANF doses of 12 to 50 pmol/minute, urinary electrolyte excretion rose in a dose related fashion and was similar in WKY and SHR. At infusions of 95 to 190 pmol/minute, the diuretic and saluretic responses were diminished in the hypertensive animals. Only the 190 pmol/minute sANF dose significantly enhanced cGMP excretion in SHR (p less than 0.05); however, in WKY urinary cGMP excretion was elevated in a dose-related fashion. At the highest sANF dose, cGMP excretion was approximately 15 times that observed in the pretreatment urine. The differences in the renal and blood pressure responses to sANF in SHR and WKY suggest that the actions of endogenous ANF may be altered in hypertension. PMID- 2987121 TI - NHLBI workshop on renovascular disease. Summary report and recommendations. PMID- 2987120 TI - Angiotensin increases inositol trisphosphate and calcium in vascular smooth muscle. AB - Angiotensin II stimulated the breakdown of phosphatidylinositol-4,5-bisphosphate (PIP2) and the generation of inositol trisphosphate (IP3) in cultured rat aortic smooth muscle cells. The decrease in PIP2 and increase in IP3 levels were rapid (measurable at 5 seconds; maximum IP3 levels at 15 seconds). The time course of these changes was comparable to that of angiotensin II-induced increases in cytosolic free calcium, as measured by the calcium-sensitive fluorescent indicator quin 2. The IP3 formation was not stimulated by the calcium ionophore A23187 (5 microM), nor were angiotensin II-induced changes in IP3 formation inhibited by the removal of extracellular calcium with EGTA. Angiotensin II appears to be capable of generating more IP3 than is required for maximal release of intracellular calcium. These data are consistent with the hypothesis that generation of IP3 plays a role in the angiotensin II-induced mobilization of calcium from intracellular storage sites in vascular smooth muscle cells. PMID- 2987122 TI - True versus immunoreactive angiotensin II in human plasma. AB - To measure specifically angiotensin-(1-8)octapeptide, peptides were extracted from 2 ml of plasma by reversible adsorption to bonded-phase silica. The angiotensin-(1-8)octapeptide was then isolated by isocratic reversed-phase high performance liquid chromatography and quantified by radioimmunoassay. The extraction recovery of 125I-angiotensin II added to 2 ml of plasma was 99 +/- 2% (mean +/- SD). The overall recovery of 5, 10, and 20 fmol unlabeled angiotensin II added to 1 ml of plasma was 80 +/- 10%. The coefficient of variation for within-assay precision was 0.06 and for between-assay precision 0.13. The detection limit was 0.4 fmol/ml. Buffer and plasma blanks were below the detection limit. Normal subjects on a free diet in supine position averaged 4.2 +/- 1.7 fmol/ml angiotensin-(1-8)octapeptide. Furosemide (40 mg p.o.) and standing increased these values to 22 +/- 7.6 fmol/ml. In four volunteers, immunoreactive "angiotensin II" (more or less angiotensin-like material) was measured serially before and after converting-enzyme inhibition (Hoe 498) with conventional Dowex extraction. At peak inhibition, plasma immunoreactive "angiotensin II" levels decreased by only 44%. In contrast, angiotensin-(1-8) octapeptide isolated by high-performance liquid chromatography completely disappeared. In hypertensive patients receiving long-term treatment with enalapril, plasma levels of angiotensin-(1-8)octapeptide fell from 2.7 +/- 0.9 to 0.9 +/- 0.3 fmol/ml (mean +/- SEM) 2 hours after the morning dose, whereas levels of immunoreactive "angiotensin II" were not significantly changed. We found that this sensitive method specifically measured angiotensin-(1-8)octapeptide and demonstrated that true angiotensin II virtually disappears during converting enzyme inhibition. PMID- 2987123 TI - Renal arachidonic acid metabolism. The third pathway. AB - Cells were isolated from the outer medulla of the rabbit kidney, primarily from the thick ascending limb of Henle's loop (mTALH). These mTALH cells are heavily invested with a cytochrome P450-linked monooxygenase that represents the third pathway by which arachidonic acid is metabolized. After cell separation, approximately 80% of the cells proved to be mTALH in origin, based on electron microscopic criteria and immunofluorescent localization of Tamm-Horsfall protein, a specific marker for mTALH cells. The specific activity of alkaline phosphatase, a marker for proximal tubular cells, decreased threefold after separation of mTALH cells from outer medullary cells, associated with a fourfold increase in the capacity of the separated mTALH cells to metabolize arachidonic acid. Incubation of mTALH cells with 14C-arachidonic acid resulted in formation of oxygenated metabolites, identified as two peaks (P1 and P2), which accounted for 30 to 40% of the recovered radioactivity. Formation of prostaglandin E2 and F2 alpha accounted for only 3 to 5%. The chromatographic retention times of P1 and P2 were different from products of lipoxygenases. An inhibitor of cytochrome P450 dependent enzymes, SKF-525A (50 microM), reduced product formation by mTALH cells by more than 70%, while induction of cytochrome P450 increased product formation. Formation of P1 and P2 by cell-free homogenates of mTALH was totally dependent on the presence of nicotinamide adenine dinucleotide phosphate, reduced form (NADPH), which suggests a NADPH-dependent cytochrome P450-linked monooxygenase pathway. Vasopressin and calcitonin (10(-10) M to 10(-7) M) stimulated release of arachidonic acid metabolites from mTALH cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987124 TI - Synthetic atrial natriuretic factor in conscious normotensive and hypertensive rats. AB - Synthetic atrial natriuretic factor (Arg-Arg-Ser-Ser-Cys-Phe-Gly-Gly-Arg-Ile-Asp Arg-Ile-Gly-Ala-Gln-Ser-Gly -Leu- Gly-Cys-Asn-Ser-Phe-Arg-Tyr-COOH [disulfide bond between cysteines]) was infused intravenously into conscious normotensive and deoxycorticosterone, one-kidney, one-clip, and two-kidney, one-clip hypertensive rats. Mean arterial pressure, urine volume, and electrolyte excretion rates were measured during a 20-minute infusion of a single dose (ranging from 0-1520 pmol/min) into each animal; 95 to 380 pmol/minute of synthetic atrial natriuretic factor maximally reduced mean arterial pressure by 20 +/- 4, -29 +/- 2, and -39 +/- 7 mm Hg in normotensive, one-kidney, one-clip, and two-kidney, one-clip hypertensive rats, respectively. In deoxycorticosterone rats, a dose of 760 pmol/minute was required to produce the largest depressor response (-58 +/- 12 mm Hg). Sodium excretion increased to 8.8 +/- 2.5 muEq/minute at 760 pmol/minute in normotensive rats, to 6.5 +/- 1.1 muEq/minute at 50 pmol/minute in deoxycorticosterone rats, and to 5.8 +/- 1.5 muEq/minute at 95 pmol/minute in one-kidney, one-clip animals. The natriuretic effect was consistently greater at all doses of synthetic atrial natriuretic factor in the two-kidney, one-clip hypertensive model, in which the maximum response was 15.3 +/- 4.7 muEq/minute at 190 pmol/minute. The changes in urine volume and excretion rates of potassium and chloride tended to parallel the increases in sodium excretion in each model. Interestingly, the maximally effective hypotensive dose of synthetic atrial natriuretic factor was different from the maximally effective natriuretic dose in all four groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987125 TI - beta-Adrenergic and cholinergic receptors in hypertension-induced hypertrophy. AB - Perinephritic hypertension was produced in dogs by wrapping one kidney with silk and removing the contralateral kidney 1 week later. Mean arterial pressure rose from 104 +/- 3 to 156 +/- 11 mm Hg, while left ventricular free wall weight, normalized for body weight, was increased by 49%. Muscarinic, cholinergic receptor density measured with [3H]-quinuclidinyl benzilate, fell in hypertensive left ventricles (181 +/- 19 fmol/mg, n = 6; p less than 0.01) as compared with that found in normal left ventricles (272 +/- 16 fmol/mg, n = 8), while receptor affinity was not changed. The beta-adrenergic receptor density, measured by binding studies with [3H]-dihydroalprenolol, rose in the hypertensive left ventricles (108 +/- 10 fmol/mg, n = 7; p less than 0.01) as compared with that found in normal left ventricles (68.6 +/- 5.2 fmol/mg, n = 15), while beta adrenergic receptor affinity decreased in the hypertensive left ventricles (10.4 +/- 1.2 nM) compared with that found in the normal left ventricles (5.0 +/- 0.7 nM). Plasma norepinephrine levels were similar in the two groups, but myocardial norepinephrine levels were depressed (p less than 0.05) in dogs with hypertension. Moderate left ventricular hypertrophy induced by long-term aortic banding in dogs resulted in elevations in beta-adrenergic receptor density (115 +/- 14 fmol/mg) and decreases in affinity (10.4 +/- 2.2 nM) similar to those observed in the dogs with left ventricular hypertrophy induced by hypertension. Thus, our results suggest that perinephritic hypertension in the dog induces divergent effects on cholinergic and beta-adrenergic receptor density. The increased beta-adrenergic receptor density and decreased affinity may be a characteristic of left ventricular hypertrophy rather than hypertension. PMID- 2987126 TI - Increased plasma renin during renin inhibition. Studies with a novel immunoassay. AB - The response of renin release to the administration of renin inhibitors cannot be studied with conventional enzymatic methods used to measure plasma renin. In the present experiments, a novel multirange enzyme-linked immunosorbent assay for human and primate renin was used to investigate the changes in plasma immunoreactive renin after renin inhibition. A potent and long-acting statine containing renin inhibitor, CGP 29 287, was injected in conscious marmosets after mild or severe sodium depletion. In mildly sodium-depleted marmosets, CGP 29 287 (0.1 mg/kg i.v.) reduced mean arterial blood pressure and completely inhibited plasma renin activity for up to 30 minutes. This response was associated with a transient increase in plasma immunoreactive renin concentration. After a dose of 1.0 mg/kg i.v., the reduction of mean arterial pressure and the complete inhibition of plasma renin activity persisted for up to 120 minutes. These effects were accompanied by a sustained increase in plasma immunoreactive renin concentration. In severely sodium-depleted marmosets, CGP 29 287 (1.0 mg/kg i.v.) induced a marked fall in systolic blood pressure and complete inhibition of plasma renin activity within 30 minutes of injection. Plasma immunoreactive renin levels increased to 257% of pretreatment values. The converting-enzyme inhibitor enalaprilat (2 mg/kg i.v.) induced a fall in systolic blood pressure of similar magnitude, which was accompanied by an increase in plasma renin activity. Levels of plasma immunoreactive renin increased to 210% of pretreatment values. Hydralazine (0.2 mg/kg i.v.) did not increase plasma renin activity or plasma immunoreactive renin levels despite a comparable hypotensive effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987127 TI - Comparison of renin and converting enzyme inhibition in sodium-deficient dogs. AB - While renin is a highly specific protease, converting enzyme has at least two principal substrates, angiotensin I and bradykinin. Changes in the rate of formation of angiotensin II or degradation of bradykinin can influence the hypotensive action of angiotensin converting enzyme inhibitors. The present study was designed to determine if there were differences in the maximal blood pressure reduction in Na-deficient dogs after angiotensin converting enzyme or renin inhibitor treatment. Five conscious dogs received 0.1, 0.5, and 1.0 mg/kg of i.v. enalaprilat, a potent angiotensin converting enzyme inhibitor, which reduced blood pressure to 75 +/- 4, 71 +/- 5, and 71 +/- 5 mm Hg. Plasma immunoreactive angiotensin II levels were reduced in a dose-related fashion to 35% of control level at the highest dose. Infusion of a maximally effective dose of a statine containing renin inhibitor (SCRIP) with the high dose of enalaprilat produced no further fall in blood pressure (68 +/- 7 mm Hg), but immunoreactive angiotensin II levels fell to essentially zero in four of five dogs. The order of drug administration was reversed in another experiment in a group of nine dogs in which SCRIP reduced plasma immunoreactive angiotensin II to 25% of control at 0.04 mg/kg/minute (n = 5), with reduction to near zero levels at higher doses. Maximal blood pressure reduction was achieved at 0.32 to 0.64 mg/kg/minute (76 +/ 4 mm Hg); 1 mg/kg of enalaprilat lowered blood pressure an additional 11 +/- 2 mm Hg (p less than 0.01) while not further decreasing immunoreactive angiotensin II levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987128 TI - Highly potent and specific inhibitors of human renin. AB - Small peptide analogues representing the C-terminal portion of angiotensin I sequence were designed as inhibitors of human renin. Among synthesized compounds, benzyloxycarbonyl (-"Z")-(1-naphthyl)Ala-His-leucinal (ES-188), Z-(1-naphthyl)Ala His-statine ethyl ester (ES-226), and Z-(1-naphthyl)Ala-His-statine 2 methylbutylamide (ES-254) markedly inhibited human and primate renins (inhibitory concentration, 50% [IC50], near 10(-7) M). These peptide analogues inhibited rabbit renin with one or two orders of magnitude less potency. They were very weak inhibitors of renins from pig, goat, dog, and rat. ES-188 had no discernible effect on cathepsin D, pepsin, or human angiotensin-converting enzyme at the concentration of 10(-4)M. ES-226 had little effect on the three enzymes at the concentration of 10(-5)M; however, ES-254 had a considerable inhibitory effect on cathepsin D (IC50 of 1.4 X 10(-5)M), pepsin (IC50 of 4.2 X 10(-5)M), and human angiotensin-converting enzyme (IC50 of 7.1 X 10(-6)M). Our results indicate that 1-naphthylalanine-containing tripeptide analogues are highly potent human renin inhibitors. PMID- 2987129 TI - Endothelium-derived vascular relaxing factor. AB - A large number and variety of compounds (acetylcholine, adenosine diphosphate, adenosine triphosphate, arachidonic acid, bradykinin, Ca2+ ionophores, calcitonin gene-related peptide, histamine, hydralazine, substance P, thrombin, and vasoactive intestinal polypeptide) have been shown to relax arterial smooth muscle indirectly. The endothelium in muscular arteries from several species appears to have receptors for these vasodilators. Binding of one of these compounds to its endothelial receptors results in the release (and presumably synthesis) of substance(s) that act on arterial smooth muscle to cause relaxation. The name endothelium-derived relaxing factor (EDRF) has been proposed for the substance or substances responsible for inhibition of contraction. Studies to determine additivity of endothelium-dependent relaxing agents and sensitivity of EDRF-mediated responses to a variety of inhibitors suggest that a single factor or a single common mechanism induces relaxation of vascular smooth muscle. Pharmacological studies have been equivocal with regard to the postulated involvement of phospholipases or arachidonic acid and to the suggestion that EDRF is an oxidative, non-cyclooxygenase product of arachidonate. Experiments on transfer of EDRF and reversal of endothelium-dependent relaxation consistently indicate that EDRF is quite labile. There is convincing evidence that EDRF activates smooth muscle guanylate cyclase, which results in an increase in intracellular cyclic guanosine 3',5'-monophosphate levels. The stimulation of guanylate cyclase by EDRF provides a valuable and sensitive parameter for studies with arteries as well as cells in culture. At present, the identity of EDRF and its role in cardiovascular homeostasis are unknown. PMID- 2987130 TI - Genetic analysis of virulence plasmid from a serogroup 9 Yersinia enterocolitica strain: role of outer membrane protein P1 in resistance to human serum and autoagglutination. AB - Enteropathogenic strains of Yersinia enterocolitica harbor a virulence plasmid (70 kilobases) which specifies, at 37 degrees C, a calcium requirement for growth, autoagglutinability, resistance to the bactericidal activity of human serum, and the expression of some outer membrane proteins (OMPs). To map the genes encoding these properties, the virulence plasmid of a serogroup 9 strain (W22708) was subjected to transposon mutagenesis. A set of 68 independent mutations was obtained in Escherichia coli by transposon Tn813 (a tnpR mutant of Tn21)-mediated cointegration with the self-transmissible R388 plasmid. The resulting cointegrates were introduced and studied in Y. enterocolitica W22708. One mutant lost the calcium dependence property. Two other mutants presented a peculiar phenotype: they grew poorly at 37 degrees C, especially in the presence of calcium. Lastly, two mutants were affected in the properties of autoagglutination and resistance to human serum. Analysis of the OMP pattern of these two mutants revealed the absence of the largest OMP, called P1 (I. Bolin, and H. Wolf-Watz, Infect. Immun. 43:72-78, 1984). Complementation of one of these mutations with the cloned structural gene of OMP P1 restored the wild-type phenotype. However, OMP P1 was not sufficient by itself to specify the serum resistance property and a rapid autoagglutination of the host. PMID- 2987131 TI - Bacterial lipopolysaccharide induction of leukocyte-derived corticotropin and endorphins. AB - Previous reports have shown that there is an endogenous opioid component associated with pathophysiological responses to endotoxin. It has been shown that these responses are alleviated by naloxone, a specific opiate antagonist. Results of another study have indicated that leukocytes may mediate some of those responses since leukocyte depletion alleviated the effects of lipopolysaccharide. In view of the above reports as well as the finding that leukocytes produce immunoreactive (ir-) endorphins and corticotropin (ACTH) when stimulated with Newcastle disease virus or ACTH-releasing factor, we postulated that leukocytes may serve as an extrapituitary source of endorphins produced in response to bacterial endotoxin. To test this hypothesis, human peripheral blood leukocytes as well as mouse spleen cells were cultured in vitro with Escherichia coli lipopolysaccharide for 48 h. The lipopolysaccharide (i.e., endotoxin) was shown to induce de novo synthesis of ir-ACTH and ir-endorphins. The leukocyte-derived ir-ACTH had a molecular weight of approximately 2,900 and demonstrated a bioactivity similar to that of pituitary-derived ACTH. The lymphocyte-derived ir endorphin comigrated with alpha- and gamma-endorphin at approximately 1,800 daltons and was shown to bind to brain opiate receptors. These findings imply that leukocyte-derived endorphins may be involved in the pathophysiological response to endotoxin. PMID- 2987132 TI - Antibiotics in pregnancy: toxicity and teratogenicity. PMID- 2987133 TI - Antibiotic prophylaxis in transurethral surgery: a comparison of sulbactam ampicillin and cefoxitin. AB - The efficacy and safety of perioperative prophylaxis with sulbactam-ampicillin or cefoxitin was compared in a prospective randomized double-blind study of 103 patients undergoing transurethral surgery. Fifty-two patients received 0.5 g sulbactam and 1 g ampicillin intramuscularly, 30 to 90 minutes prior to surgery, followed by the same dose administered intravenously every eight hours for a total of three additional doses over 24 hours. Fifty-one patients received 1 g of cefoxitin administered according to the same schedule as sulbactam-ampicillin. The incidence of urinary tract infection during hospitalization was 8% in the sulbactam-ampicillin group and 4% in the cefoxitin group. One month postoperatively the incidences were 3% and 5%, respectively. There was no significant difference between the groups in incidence of fever or length of postoperative hospital stay. Both drugs were well tolerated. No side effects were seen other than a mild skin rash in one patient and diarrhea in two patients. Sulbactam concentrated in prostatic tissue, and ampicillin together with sulbactam was found in concentrations above the minimal inhibitory concentration of most bacteria causing postoperative urinary tract infection. It is concluded that sulbactam-ampicillin and cefoxitin are equally effective and safe in preventing postoperative urinary tract infection in transurethral surgery. PMID- 2987134 TI - Erythrocyte pyrimidine 5'-nucleotidase activity in workers exposed to lead, mercury or cadmium. AB - The erythrocyte pyrimidine 5'-nucleotidase activity (P5N) was measured in workers exposed to lead, cadmium or inorganic mercury and the effect of lead on this enzyme activity was compared with that on the classical biological parameters (ALAD, ZPP, ALAU), reflecting the interference of this metal with the heme biosynthesis pathway. P5N is inhibited by lead but not by cadmium and mercury. The correlation coefficient between log P5N and lead in blood (PbB) amount to 0.79 (n = 278; PbB ranging from 4.9 to 73.5 micrograms/100 ml). The response of P5N to lead closely parallels that of ALAD. PMID- 2987135 TI - Production of beta-human chorionic gonadotrophin by human squamous carcinoma cell lines. AB - Eight out of 9 human squamous carcinoma cell lines ectopically secrete the beta subunit of human chorionic gonadotrophin (beta-HCG) in amounts detectable by radioimmunoassay. In contrast, both normal and SV40-transformed keratinocytes show no such evidence of beta-HCG secretion. Ectopic beta-HCG is identical in its gel exclusion and high-performance liquid chromatographic properties to placental beta-HCG and, in radioimmunoassay, yields a dilution curve parallel to that of placental beta-HCG. Dibutyryl cyclic AMP, but not sodium butyrate, stimulates beta-HCG secretion by the one human cell line examined, LICR-LON-HN-5, suggesting that ectopic production by these cells is similar to trophoblastic beta-HCG secretion, and differs from most other non-trophoblastic cell lines previously examined. PMID- 2987136 TI - Selective cytotoxicity of murine monoclonal antibody LAM2 against human small cell carcinoma in the presence of human complement: possible use for in vitro elimination of tumor cells from bone marrow. AB - LAM2 is a murine IgM monoclonal antibody (MAb) which binds strongly to the cell membrane of human lung small-cell carcinoma (SCC) and squamous-cell carcinoma but not to normal bone-marrow cells. The cytotoxicity of this antibody in the presence of human complement was investigated in vitro by chromium release and clonogenic assays. The optimal treatment conditions included incubation with antibody for 30 min at 37 degrees C followed by 3 additions of human complement 30 min apart. Cell lysis ranged from 94 to 98% in 4 SCC cell lines at antibody dilutions of 1:100: a lower level of lysis (60%) occurred in a lung squamous-cell carcinoma cell line. The cytotoxic effect was strictly complement-dependent. No cytotoxic effect was seen with other human cell lines including lung adenocarcinoma, lung large-cell carcinoma, myeloid leukemia, and lymphoblastic leukemia. No lysis was seen with nucleated marrow cells from healthy volunteers. Normal marrow cells in excess did not inhibit SCC cell lysis. Incubation with antibody and complement resulted in a 100-fold reduction of colony formation of SCC cells, but did not reduce the number of colonies of marrow-cell precursors, including CFU-GEMM, BFU-E, and CFU-C. The selective cytotoxicity of LAM2 antibody to SCC, but not to normal bone-marrow cells, suggests that this antibody may be useful for the in vitro elimination of SCC cells from the bone marrow. PMID- 2987137 TI - Chronic alpha-adrenoceptor blockade with trimazosin in congestive heart failure. AB - We evaluated chronic adjunctive alpha-1 receptor blockade with trimazosin in congestive heart failure. This agent produced hemodynamic effects consistent with venodilation (reduced left ventricular volume and filling pressure with increased left ventricular ejection fraction), but only during exercise. Resting hemodynamic parameters and exercise duration were not significantly altered by chronic alpha-1 receptor blockade. PMID- 2987138 TI - Renin-aldosterone axis in ethanol intoxication: effect of A II infusion. AB - Plasma renin activity (PRA) is increased while plasma aldosterone is not, during moderate ethanol intoxication. To elucidate mechanisms behind this dissociation of renin-aldosterone nexus, six healthy males were given angiotensin II (4 ng kg 1 min-1) by i.v. infusion following ingestion of ethanol (1.2 g kg-1 body weight). Prior to angiotensin II (A II) infusion, PRA rose and plasma aldosterone declined. A II infusion caused a roughly 3-fold increase of plasma aldosterone both in ethanol intoxication and in control experiments, and a transient suppression of PRA. Plasma renin substrate, cortisol, and angiotensin converting enzyme (ACE) remained unchanged. The decrease of serum potassium or the rise of Na+/K+ ratio in ethanol intoxication may explain the failure of the adrenal cortex to respond with aldosterone release to endogenous angiotensin II. However, the pressor dose of A II infused obviously overcame the blunting effect of ethanol on aldosterone release previously reported by us. Blood pressure, both diastolic and systolic, increased similarly during ethanol and control experiments in response to A II infusion, except in one subject, who during ethanol intoxication experienced a paradoxical fall in blood pressure while reacting normally to A II infusion without ethanol. PMID- 2987140 TI - Drug treatment effectiveness: the case of racial and ethnic minorities in America -some research questions and proposals. PMID- 2987139 TI - Effect of oral charcoal and urine pH on dextropropoxyphene pharmacokinetics. AB - The effects of orally given activated charcoal, sodium bicarbonate and ammonium chloride on the pharmacokinetics of dextropropoxyphene were studied in six volunteers in a randomized, cross-over study. Serum and urine concentrations of dextropropoxyphene and norpropoxyphene were determined by GLC up to 72 h. Activated charcoal (50 g) given 5 min after dextropropoxyphene hydrochloride (130 mg), reduced its absorption by 97-99%. When given in repeated doses from 6 h on, 50 g followed by 12.5 g at 6 h intervals, charcoal significantly shortened the terminal serum half-life of dextropropoxyphene from 31.1 +/- 4.2 h to 21.2 +/- 3.1 h (p less than 0.05) and that of norpropoxyphene from 34.4 +/- 2.5 h to 19.8 +/- 3.4 h (p less than 0.001), and reduced their excretion into urine. The cumulative urinary excretion of unchanged dextropropoxyphene was increased 6-fold by acidification and reduced to 1/20 by alkalinization of urine, but the excretion of norpropoxyphene was much less dependent on urinary pH. However, the cumulative excretion of dextropropoxyphene and norpropoxyphene even into acidic urine accounted for less than 25% of the dose during 72 h. Because urinary pH has a great influence on the ratio of urinary versus serum dextropropoxyphene concentrations, pH should be taken into consideration, when the clinical significance of its concentration in urine is evaluated. Activated charcoal in high doses effectively prevents the absorption of that fraction of dextropropoxyphene which is in the stomach at the time of charcoal administration. Given in repeated oral doses, charcoal increases to some extent the rate of elimination of dextropropoxyphene and norpropoxyphene, probably by interrupting their enterohepatic or enteroenteric circulation. PMID- 2987141 TI - The X-linked hypophosphatemic vitamin D resistant rickets: old and new concepts. PMID- 2987142 TI - Molecular analysis of mutagenesis in mammalian cells. AB - Mammalian cells are constantly facing various types of mutagens. However, due to the high complexity of the cell genome, the molecular analysis of mutagenesis has not yet been possible. Therefore, we have used simian virus 40 (SV40) as a biological and molecular probe to characterize mutagenesis at the nucleotide level. By using a reversion assay from a temperature-sensitive phenotype towards a wild-type phenotype, we have analysed mutagenesis induced by u.v.-light and by apurinic sites (Ap sites). We report here experiments allowing us to quantify and to compare the mutagenic efficiency of various DNA lesions measured on the SV40 genome. The Ap sites are very mutagenic in this type of assay. The molecular analysis of u.v.-induced mutagenesis reveals that mutations correspond to single base-pair substitutions always located opposite Py-Py lesions. The mutations are almost equally distributed between transition and transversion types, and between the 5' and the 3' side of the Py-Py targets. These results demonstrate for the first time in animal cells the existence of targeted mutations induced by u.v. light. We propose therefore, the use of SV40 as an efficient biological and molecular probe for assaying mutagenic pathways in mammalian cells. PMID- 2987143 TI - Neurite formation modulated by nerve growth factor, insulin, and tumor promoter receptors. AB - Until recently, nerve growth factor could be considered the only neurotrophic factor with an established physiological role. We discuss the emerging evidence indicating that the insulinlike factors may constitute a family of related neurotrophic proteins, and the observations suggesting that the receptor for the phorbol ester tumor promoters is closely associated with neuronal differentiation. The emphasis of the discussion is placed on neurite formation under multiple modulation by insulinlike factors, nerve growth factor, and tumor promoter receptors in sensory, sympathetic and human neuroblastoma cells. PMID- 2987144 TI - The epidermal growth factor (EGF) receptor gene studied using gene transfer. PMID- 2987145 TI - The platelet-derived growth factor receptor. PMID- 2987146 TI - Characteristics of the nerve growth factor receptors and a nerve growth factor receptor--nerve growth factor covalent complex. AB - Nerve growth factor is a polypeptide hormone that is required for the normal growth and development of the embryonic sensory and sympathetic nervous systems. On these cells, there are two different receptors for the nerve growth factor. Recently, these receptors have been isolated from three cell types and shown to have essentially the same binding characteristics. Molecular weights for receptors from two of these cell types, embryonic sensory and rat pheochromocytoma cells, have been determined. In addition, the formation of a covalent nerve growth factor, nerve growth factor receptor complex, has been investigated on embryonic sensory and sympathetic neurons. The formation of this covalent complex containing 125I-beta nerve growth factor is prevented by the addition of excess unlabeled nerve growth factor or by the addition of sodium fluoride and dinitrophenol. This complex forms at 4 degrees, 22 degrees, and 37 degrees, indicating that it is occurring on the cell surface. A disulfide bond is involved in the formation of this covalent complex. PMID- 2987147 TI - Mammary transfer of vitamin D3 in the rabbit doe. AB - Mammary transfer of vitamin D3 from a single intravenous injection containing 100 000 IU vitamin D3 in ethanol, was studied in lactating rabbit does. Significant amounts of the injected vitamin D3 appeared in the milk of does. The highest concentration was observed in the first milk sampling (24 h after administration) which was followed by a decreasing pattern in the five postinjection days. Vitamin D3 concentration in the neonate tissues were smaller than in their dams. In the maternal tissue the highest concentration occurred in the liver, whereas muscle had the lowest value. Among the various neonatal tissues liver contained the highest amounts although in the other tissues appreciable amounts of vitamin D3 were recovered. PMID- 2987148 TI - Plasma vitamin D3 response in cattle and sheep exposed to ultraviolet radiation. AB - Plasma levels of vitamin D3 were determined in a group of 10 cows housed indoors and exposed during the winter to ultraviolet irradiation (UVR). Exposure to UVR was associated with marked rises in plasma vitamin D3 concentrations. The pattern and magnitude of the plasma vitamin D3 response to UVR exposure in sheep raised in total confinement was also investigated. Exposure to sun lamps produced a significant increase in sheep plasma vitamin D3 concentrations. There was no significant difference in the plasma levels of vitamin D3 due to length of time of exposure to UVR. PMID- 2987149 TI - Stopped flow ESR study of the action of carbohydrates on ascorbic acid oxidation in aqueous solution. AB - The influence of added carbohydrate on the decay kinetics of the ascorbic acid free radical generated in deaerated aqueous solutions by the action of potassium ferricyanide on ascorbic acid has been studied. Addition of carbohydrate (less than or equal to 0.5M) generally leads to a small decrease in radical decay rate. In our system this probably arises from reduced reactant diffusion in solutions of increased viscosity rather than the preferential chelation previously suggested to explain inhibition in the copper (II) catalyzed oxidation. The ascorbic acid free radical shows negligible tendency to react with glucose, sorbitol or maltose. The decrease in its decay rate observed on addition of carbohydrate is probably caused by the increase in solvent viscosity. PMID- 2987150 TI - The outer horizontal cell of the frog retina: morphology, receptor input, and function. AB - The outer horizontal cell (OHC) of the frog Rana pipiens was studied by light and electron microscopy of Golgi-stained and horseradish peroxidase-injected cells. Responses of OHCs were recorded with intracellular electrodes. The OHCs are probably axonless cells. Their dendritic terminals are lateral processes at synaptic ribbons and are involved frequently in reciprocal invaginating contacts with receptors, ie, contacts characterized by invagination of receptor membrane deeply into the horizontal cell process. Two classes of OHC were distinguished on the basis of size and receptor contacts. Small OHCs (dendritic area about 5,000 micron2) contact all classes of receptor. These cells were not successfully penetrated with microelectrodes. Giant OHCs (dendritic area about 30,000 micron2) apparently contact only blue-sensitive rods and red-sensitive cones. They generate chromaticity or C-type responses, hyperpolarizing to short and depolarizing to long wavelength light. PMID- 2987151 TI - Study of vitreous liquifaction by NMR spectroscopy and imaging. AB - The vitreous gel is primarily composed of collagen, hyaluronic acid, and water (98-99%) and can break down into a liquid state devoid of collagen. The liquifaction of vitreous gel that occurs with age and in certain other disease states is believed to be important in the pathogenesis of retinal tears and detachments. The authors report measurements of water proton relaxation times and water proton nuclear magnetic resonance (NMR) imaging to study the process of vitreous liquifaction in extracted vitreous and in the intact bovine eye. A comparison is made between macroscopic viscosity, longitudinal (T1) and transverse (T2) relaxation times obtained on an NMR spectrometer and proton NMR images. Vitreous liquifaction as measured by a decrease in macroscopic viscosity resulted after treatment of vitreous with collagenase and to lesser degree with hyaluronidase. A shortening of relaxation times accompanied the drop in viscosity. An area of brightness or increased proton signal intensity corresponding to a focus of liquifaction was seen by NMR imaging only after injection with collagenase. The authors believe NMR imaging to be a useful new diagnostic modality by which vitreous liquifaction can be studied in the intact eye. The vitreous provides a new model for studying changes in proton relaxation times of protein solutions in biologic systems. PMID- 2987152 TI - Atypical Verner-Morrison syndrome. PMID- 2987153 TI - Problems of etiology in femoral neuropathies. AB - 29 cases of femoral mononeuropathy are reported. While the clinical features of the femoral neuropathy are easily identified, the etiology is often hard to establish. The cases reported tend to fall into three general categories: 1) cases without major diagnostic difficulties (e.g. diabetic neuropathy); 2) those in which the definite diagnosis results from combined evidence of laboratory and instrumental data (degenerative changes in the lumbar spine, compressions, entrapments, etc.); 3) those in which the negative result of the investigations prevents a positive diagnosis and hence a presumptive etiology (spondylosis, inflammatory process, ischemia of the nerve) may be formulated. Attention is drawn to the favorable course of the condition in the patients of this group. PMID- 2987154 TI - Intraductal papillomas: diagnostic and surgical procedures. AB - A series of 98 patients with spontaneous nipple discharge, is reported. Diagnosis was based on: clinical examination, cytology of breast secretion, mammography and galactography. Surgical resection was recommended in the following cases: galactographic evidence of intraductal papilloma or papillomatosis, dubious or positive cytology, persisting hemorrhagic or sero-hemorrhagic secretion. The injection of vital staining before the operation allowed the precise location of the lesion. In the group of patients studied ten cases of ductal carcinomas (5 in situ and 5 smaller than 1 cm), 4 cases of atypical intraductal hyperplasia, 13 cases of solitary papilloma and 22 cases of multiple papillomatosis were diagnosed. PMID- 2987155 TI - The effect of age on beta-adrenergic function in man: a review. AB - The structure and function of the beta-adrenergic adenylate cyclase complex in the elderly is reviewed. The function of the beta-adrenergic receptor in man is modulated by levels of circulating catecholamines, noncatecholamine hormones, drugs, disease, and age. Although a number of clinical observations demonstrate an age-related decrease in catecholamine responsiveness, the molecular basis of this phenomenon is unknown. Simple reduction in beta-receptor number does not appear to explain age-associated loss of catecholamine responsiveness. Recent investigations from our laboratory employed salbutamol-induced rise in plasma cyclic AMP (cAMP) levels to study the molecular basis for this phenomenon. In young individuals there was a threefold increase in plasma cAMP levels after salbutamol infusion. In older subjects only a 50% rise in plasma cAMP levels was observed. These results suggest that the basis for reduced catecholamine responsiveness in the elderly is due to a defect in the peripheral beta-receptor linked adenylate cyclase complex. The finding of reduced beta-adrenergic stimulated adenylate cyclase activity in the aged prompted us to determine the specificity of this decline by measuring the activity in older and younger individuals of another hormone-sensitive adenylate cyclase. However, no effect of subject age was observed on glucagon-sensitive adenylate cyclase activity, suggesting that the blunted response of the beta-receptor adenylate cyclase complex in the elderly represents a specific loss of function and is not due to a general age-associated decline in hormone-stimulated cyclase function. Specific molecular defects which could account for decline in beta-adrenergic responsiveness in the elderly are discussed. PMID- 2987157 TI - [Hand-foot-mouth disease]. AB - A case of hand-foot-and-mouth disease is reported in a 20-year-old female patient. Infection by Coxsackie A 16-virus was diagnosed by serology. PMID- 2987158 TI - [Eccrine spiradenoma]. AB - Eccrine spiradenomas are rare and usually painful tumours, but most textbooks and reviews contain no photographs of typical cases. Therefore we report on this tumour and document our report with clinical photographs. PMID- 2987156 TI - Immunomodulatory activity of anti-atherogenic drugs: effects on blastogenesis, humoral response, delayed hypersensitivity and passive anaphylaxis by immune complexes. AB - The effect of several anti-atherogenic drugs (ticlopidine, nicotinic acid and etofibrate) on immune responses and immune complex anaphylaxis has been studied in mice. All the drugs enhanced the activation by concanavalin A, phytohemagglutinin, and lipopolysaccharide of lymphocytes taken from treated animals. Contact hypersensitivity to trinitrochlorobenzene was inhibited by similar treatments with the same drugs, possibly through inhibition of the efferent phase of the reaction. Nicotinic acid produced a slight enhancement of antibody responses to sheep erythrocytes, whereas etofibrate inhibited the response at the highest dose studied. In addition, treatment with these drugs variably protected the mice from anaphylactic shocks induced by immune complexes. Marked protection was also observed using the antiserotoninic, cyproheptadine. These results indicate that drugs used to prevent atherogenic processes modulate different proliferative and effector immunological reactions. PMID- 2987159 TI - Beneficial short-term effects of unprocessed wheat bran on lipid and glucose metabolism in man. AB - Supplementation of the normal diets of seven healthy 18-22-year old male and female volunteers with 0.15 g unprocessed wheat bran/kg body weight/d for 6 weeks increased fibre consumption by 35 per cent without noticeably affecting the intake of other major nutrients. Fasting concentrations of plasma total cholesterol, triglyceride and glucose were unchanged by wheat bran supplementation, whereas HDL-cholesterol was increased and LDL-cholesterol decreased by 46 per cent and 25 per cent respectively after 6 weeks. Glycosylated haemoglobin was decreased by 18 per cent, 27 per cent and 45 per cent after 2, 4 and 6 weeks. The results illustrate the potential value of consuming relatively small amounts of unprocessed wheat bran for the promotion of health and treatment of certain metabolic diseases in man. PMID- 2987160 TI - Quantitative histochemical assessment of regional differences in hepatic glucose uptake and release. AB - As a further step in the investigation of the heterogeneity of liver cells in general and regionality of glucose metabolism in particular, requirements for isolation of appropriate tissue samples were defined and procedures for measurement of the biochemical parameters responsible for glucose uptake and release developed and tested. By using enzymatic cycling for chemical amplification, in conjunction with the oil-well technique, sufficient analytical sensitivity was provided to assay samples averaging 20 ng dry weight. Microchemical data on the distribution of glucokinase and glucose-6-phosphatase and of their substrates, glucose and glucose-6-P, were used to, first calculate in vivo rates of these catalytic steps by means of the Michaelis-Menten equation, and then, to determine the direction and rate of net glucose flux, as well as, the rate of substrate cycling between glucose and glucose-6-P. Calculations from the results indicated a reciprocal distribution of in vivo glucokinase and glucose-6-phosphatase velocities, as well as, sex-specific differences. The distribution of in vivo activities results in a spatial separation of these antagonistic steps. Separation is incomplete, but nevertheless appears to lead to regionally different rates in futile substrate cycling. Glucose gradients permit differentiation between net glucose uptake and release and were, therefore, used as a test of the validity of the calculations of in vivo activities. The observed discrepancies between glucose gradients and calculated in vivo enzyme activities illustrate the power of this approach: it provides a way to compare changes in glucose along the sinusoid with what would be predicted from the levels of enzymes which liberate and tie up glucose and of their respective substrates. PMID- 2987162 TI - Immunocytochemical detection of triphosphoinositide in vestibular hair cells. AB - Triphosphoinositide (TPI), an aminoglycoside receptor and a possible regulator of cationic permeation through its ability to bind with Ca++, was localized by the protein-A gold technique in vestibular sensory epithelia using an antibody highly specific to TPI. TPI was detected on the stereocilia, kinocilia, and cuticular plate of hair cells, and in the reticular membrane of supporting cells. The cilia of hair cells are damaged by aminoglycosides at a relatively early stage of toxicity. Ca++-regulated bioactivity in this area is probably involved. PMID- 2987161 TI - Immunocytochemical localization of vesicular stomatitis virus proteins N and NS with monoclonal antibodies. AB - The purpose of this paper is to describe the immunocytochemical-localization of N and NS nucleocapsid proteins of vesicular stomatitis virus in the cells throughout the infectious cycle. N protein was detected in the cytoplasm at 2 h after infection and formed small cytoplasmic clusters which progressively increased in size and number. At 5-6 h, it formed large cytoplasmic inclusions. NS protein was detected in the cytoplasm a little later than N protein and showed almost the same immunostaining pattern. However, diffuse background staining of NS protein was identified throughout the cytoplasm by double immunostaining methods. At electron microscopic level, N protein was mostly granular and occasionally organized in strands at 2-3 h. At 5-6 h, numerous immunostained reaction products were organized in strands. The reaction products of NS protein were almost the same as those of N protein with the exception that diffuse background staining was observed. Cos cells, transfected with SV40 vector containing N gene obtained by recombinant DNA technique, showed clusters of N protein, but virtually no strand at electron microscopic levels. The rapid freezing and deep-etching replica method demonstrated that loosely coiled VSV genome coated with N protein was localized on cytoplasmic sides of cell membranes in the infected cells. These results showed that complete virus genome replication was needed for strand formation of N and NS proteins and suggested that they were bound to VSV genomes in the infected cells. PMID- 2987163 TI - Prevention of penetration hindrance in cerium-based glucose-6-phosphatase cytochemistry by freezing tissue in melting nitrogen. AB - The demonstration of the ultrastructural localization of glucose-6-phosphatase in rat liver is hampered by penetration problems of the medium as appears from ultrathin cross-sections of vibratome sections. Besides the plasma membrane, also the cytoplasm forms a serious barrier for the penetration of the medium constituents. Prolonged preincubation for as long as 48 h at 4 degrees C in the complete incubation medium could not prevent the penetration hindrance. However, when employing 30 micron vibratome sections from tissue blocks that were frozen in melting nitrogen, the penetration hindrance was prevented. PMID- 2987164 TI - Computed tomography in the staging of small cell lung cancer: implications for combined modality therapy. AB - Computed tomography of the thorax and abdomen, from the thoracic inlet to the renal hila, was performed as part of initial staging in 51 patients with small cell carcinoma of the lung (SCCL). The computed tomographic (CT) scans were repeated after completion of chemotherapy, as part of routine restaging and assessment of response to therapy. To identify the ways in which CT scanning uniquely benefited evaluation of initial disease extent in comparison to other diagnostic studies exclusive of CT scan, all diagnostic and clinical data were reviewed. CT scan identified more advanced intrathoracic disease than chest radiography in 82% of patients. Mediastinal node involvement not appreciated by chest radiography was seen in 61% of patients. Adrenal and retroperitoneal node involvement, not suspected by other studies, was identified by CT scan in 31% and 12% of patients, respectively. Thirty percent of the patients staged as limited disease (LD) were advanced to extensive disease (ED) by CT scan findings. While confirmation, by biopsy, of positive CT findings was not consistently accomplished, restaging CT scans provided indirect confirmation by displaying improvement or worsening that correlated with disease regression or progression. Thoraco-abdominal CT scanning more accurately identifies the extent of small cell carcinoma than other imaging procedures. This has important implications for reporting results by extent of disease. In addition, CT more accurately identifies the magnitude of intrathoracic primary and nodal tumors, which may influence the choice and conduct of local treatment--surgery and/or radiation therapy--in combination with systemic chemotherapy. PMID- 2987165 TI - Tumor radiosensitization with concomitant bone marrow radioprotection: a study in mice using diethyldithiocarbamate (DDC) under oxygenated and hypoxic conditions. AB - We have established, both in vitro and in vivo, that Diethyldithiocarbamate (DDC) protects mammalian cells from radiation. The in vivo protection, when non-toxic concentrations of DDC are present one-half hour before irradiation, is reflected by a dose modification factor (DMF) of 1.9 based on LD50/30 and 1.5 using survival of CFUs as an endpoint. Further experiments in vivo have extended our knowledge to the differential radioprotective effects of DDC on the bone marrow of animals breathing room air compared to a 5.5% oxygen in nitrogen mixture. The DMF (LD50/30) for DDC in air breathing animals, previously established as 1.9, can be contrasted with a DMF, obtained in the present study, of 1.2 for animals irradiated in the hypoxic state. Moreover the DMF (CFUs survival) previously established at 1.5 for air breathing animals, can be compared to a value of 1.3, obtained in the present study, for mice irradiated under hypoxic conditions. Modification of the dose response by DDC, for both bone marrow and tumor, was also examined in animals bearing a RIF sarcoma. Although protection of the bone marrow was confirmed (DMF = 2.1), the striking finding was that the tumor cells were sensitized, in both air breathing and nitrogen breathing animals, by the addition of DDC one-half hour before the radiation exposure. Moreover, the tumor radiosensitization, a factor greater than 2, in air breathing animals, appeared to be independent of dose (D0 = 200 rad, with or without DDC). The tumor radiosensitization was even more marked in the nitrogen breathing mice, in which a factor of 10 difference in survival was noted, together with a tendency towards greater sensitization at radiation doses in the clinical range. The results, demonstrating bone marrow radioprotection by DDC (aerobic greater than hypoxic) with concomitant tumor radiosensitization (hypoxic greater than aerobic) strongly suggest a large therapeutic gain factor (TGF) which could be further explored in a clinical setting. PMID- 2987167 TI - Evaluation of the radial immunodiffusion enzyme assay for the detection of antibodies to pseudorabies virus. AB - The validity of radial immunodiffusion enzyme assay (RIDEA) as a diagnostic test for antibodies to pseudorabies virus (PRV) in porcine serum was determined. Serum samples from sows and offspring were tested for the presence of antibodies to PRV, using both the RIDEA and the PRV serum-neutralization (SN) test. Overall sensitivity and specificity of the RIDEA done on serums from the sows were 95.7% and 95.2%, respectively. This sensitivity compares with 97.3% sensitivity of the SN test of the same serums. In 658 swine serum samples from routine submissions to the University of Missouri-Columbia Veterinary Diagnostic Laboratory that were tested by the RIDEA, the calculated sensitivity and the specificity were 94.3% and 98.9%. The RIDEA and SN test were equally sensitive (99.0%) to detect antibodies resulting from infection with a field strain of virus. They had reduced sensitivity (RIDEA, 91.7%; SN test, 95.2%) in tests of serums from vaccinated sows. For the detection of passively transferred antibodies in young pigs, sensitivity of the RIDEA was 76.1%, and specificity was 100%. In all instances, RIDEA was 100% sensitive at SN titers of 1:16 or greater. In testing serum samples of swine after field virus infection, sensitivity and specificity of the RIDEA approximated those of the SN test. This reliability, together with its ease of performance, makes the RIDEA an ideal field test in programs to detect PRV-infected herds and in programs designed to free herds of PRV infection. PMID- 2987168 TI - Visna-like disease in a Minnesota sheep. AB - Visna-like disease was diagnosed in a 4-year-old Mouflon ram from a Minnesota flock. The diagnosis was based on the unique distribution pattern of histologic lesions in the brain. Although visna is rarely diagnosed in the United States, this case illustrates that it should not be overlooked in differential diagnosis of neurologic disorders in sheep. PMID- 2987166 TI - Measurement of WR-2721, WR-1065, and WR-33278 in plasma. AB - Alkaline phosphatase was used in developing two new assays for WR-2721; one involved simple spectrophotometric titration with Ellman's reagent and the other was based upon HPLC analysis of a monobromobimane derivative. Both methods gave acceptable results when applied to whole plasma. Assays for WR-1065 and disulfide forms of the drug were also developed but were found to give unreliable results with whole plasma owing to apparent rapid reaction of these drug forms with plasma constituents. The methods were applied to the analysis of blood samples in rat 0-70 min after i.v. injection of WR-2721. PMID- 2987169 TI - Localization of cellular retinol-binding protein and cellular retinoic acid binding protein in the rat testis and epididymis. AB - The distribution of cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein (CRABP) in rat testis and epididymis was examined by the peroxidase-antiperoxidase immunolocalization technique. In the testis, cellular retinol-binding protein was localized exclusively in the Sertoli cells. Staining varied with the stages of the seminiferous epithelium cycle and was maximal prior to the maturation divisions. Cellular retinoic acid-binding protein was localized exclusively in the germinal cells in the adluminal compartment. The results suggest that retinoic acid may be the retinoid form used by the germinal cells, and that Sertoli cells may use the cellular retinol-binding protein to transfer retinol from the basal to the adluminal compartment. In the epididymis, cellular retinol-binding protein was localized in the cytoplasm and stereocilia of the principal cells in the proximal caput epididymidis, while cellular retinoic acid binding protein was localized in the spermatozoa and the stereocilia of the principal cells throughout the epididymis and in the epithelial cells of the distal vas deferens. Sperm staining intensity decreased from the initial segment to the cauda. The presence of high levels of cellular retinol-binding protein in the epithelial cells and high levels of cellular retinoic acid-binding protein in the spermatozoa of the caput epididymidis, known to be involved in the synthesis and secretion of factors necessary for sperm maturation, suggests that vitamin A may have a role in this process. PMID- 2987170 TI - A new 5'-nucleotidase inhibitor, nucleoticidin. I. Taxonomy, fermentation, isolation and biological properties. AB - A new 5'-nucleotidase inhibitor named nucleoticidin was produced by Pseudomonas sp. YM-3229G. It was isolated from a fermentation broth by trichloroacetic acid extraction, ethanol precipitation and Dowex 1 and DEAE-52 column chromatography. It inhibited 5'-nucleotidase activity of snake venom and rat liver membrane. It also showed antitumor activity against solid type Sarcoma 180. PMID- 2987171 TI - A new 5'-nucleotidase inhibitor, nucleoticidin. II. Physico-chemical properties and structure elucidation. AB - A novel 5'-nucleotidase inhibitor, named nucleoticidin, was isolated from a fermentation broth of Pseudomonas sp. The molecular weight was estimated by gel filtration to be over 1,000,000. Nucleoticidin is composed of D-glucose and D mannose at a molar ratio of 1.7 to 1.0. Combined analyses using chemical and physico-chemical methods, such as gas liquid chromatography and mass fragmentography, revealed that nucleoticidin has a structural unit with mannosyl residues at the terminal of a (1----4) linked D-glucosyl main chain with beta configuration. PMID- 2987172 TI - The structure of amphotericin A. II. The complete structure of the antibiotic. AB - The structure of amphotericin A without the configuration of asymmetric carbon atoms has been elucidated. The stereochemistry of the sugar moiety has been determined. On the basis of homoscalar correlated 2D 1H spectra. of amphotericin A the position of the hemiketal moiety has been located, and the chemical shifts of all the protons in the antibiotic molecule have been determined. PMID- 2987173 TI - Binding of [14C]tuberactinomycin O to ribosomes from Mycobacterium smegmatis. PMID- 2987174 TI - Pharmacokinetics and bactericidal activity of sultamicillin in infants and children. AB - The pharmacokinetics of sultamicillin and ampicillin suspensions were studied in 20 infants and children 8 months to 69 months of age (mean age, 27 months). Mean peak plasma concentrations of ampicillin and sulbactam occurred at 90 minutes after administration of 42.5 mg of sultamicillin (25 mg of ampicillin/kg and 17.5 mg of sulbactam/kg) per kg to fasting and non-fasting patients. Co-administration of milk usually resulted in higher concentrations of ampicillin and sulbactam, however, the differences in the AUC values between the fasting and fed groups were not statistically significant. Sultamicillin and ampicillin were administered in cross-over fashion to ten children. Plasma concentrations of ampicillin after 42.5 mg of sultamicillin per kg were greater at 20, 40, and 60 min than those after 25 mg of ampicillin per kg alone and the AUC was 39% larger in subjects who received sultamicillin than in those who received ampicillin. Plasma bactericidal activity against a non-beta-lactamase producing Haemophilus influenzae strain was similar for children who were given sultamicillin or ampicillin. Against a beta-lactamase-producing Haemophilus strain the median bactericidal titres were 1:8 at 40, 60 and 90 min after sultamicillin and less than 1:2 at the same intervals after ampicillin. PMID- 2987175 TI - Work performance in iron deficiency of increasing severity. AB - The effect of iron deficiency on work capacity was studied in groups of rats that had received diets with iron contents ranging between 9 and 50 mg/kg diet from 3 to 6 wk of age. Maximal O2 consumption (VO2max) declined only 16% with a decrease in hemoglobin (Hb) from 14 to 8 g/dl and fell sharply only below a Hb of 7 g/dl. Duration until exhaustion in a treadmill exercise of submaximal intensity (endurance) showed no significant depression between a Hb of 14 and 10 g/dl. However, endurance declined abruptly by 73% between a Hb of 10 and 8 g/dl. The VO2max results are in accord with known compensatory mechanisms that help to maintain delivery of O2 to tissues until anemia becomes severe. The sharp fall in endurance with relatively mild iron deficiency suggests a lack of similarly effective compensations for decreased oxidative capacity of muscle. PMID- 2987177 TI - Effect of muscle length on electromyogram in a canine diaphragm strip preparation. AB - The relationship between diaphragm electromyogram (EMG), isometric force, and length was studied in the canine diaphragm strip with intact blood supply and innervation under three conditions: supramaximal tetanic (100 Hz) phrenic nerve stimulation (STPS; n = 12), supramaximal phrenic stimulation at 25 Hz (n = 15), and submaximal phrenic stimulation at 25 Hz (n = 5). In the same preparation, the EMG-length relationship was also examined with direct muscle stimulation when the neuromuscular junction was blocked. EMG from three different sites and via two types of electrodes (direct or sewn-in and surface) were recorded during isometric contraction at different lengths. Direct EMGs were recorded from two bipolar electrodes sutured into the strip, one near its central end and the other near its costal end. A third EMG electrode configuration summed potentials from the whole strip by recording potentials between central and costal sites. Surface EMGs were recorded by a bipolar spring clip electrode that made contact with upper and lower surfaces of the muscle strip with light pressure. In all conditions of stimulation with different types of electrodes, all EMGs decreased significantly (P less than 0.05) when muscle length was changed from 50 to 120% of resting length (L0). Minimal and maximal force outputs were observed at 50 and 120% of L0, respectively, in all experiments. The results of this study indicated that the muscle length is a significant variable that affects the EMG recording and that the diaphragmatic EMG may not be an accurate reflection of phrenic nerve activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987176 TI - Patellar tendon matrix changes associated with aging and voluntary exercise. AB - Male rats maintained under constant environmental conditions were randomly assigned to nonrunner (NR) and voluntary exercise (R) groups. At 9 mo, voluntary exercise significantly increased muscle cytochrome c concentration and citrate synthase activity. Also, at the same age, R animals had significantly greater glycosaminoglycan concentration than NR, but no changes in dry weight and collagen concentration were significant. By age 28 mo, the R groups had reduced daily running by 70%, and elevation of tendon glycosaminoglycans relative to NR animals was no longer statistically significant. A similar trend was noted for muscle mitochondrial markers. Aging significantly decreased tendon glycosaminoglycans and increased collagen concentration. Although aging reduced the total amount of voluntary exercise, the concentration of tendon glycosaminoglycans in 28-mo-old runners was equivalent to levels in 9-mo-old sedentary rats, suggesting that voluntary exercise slowed the decline in galactosamine-containing glycosaminoglycans with aging. PMID- 2987178 TI - Influence of exercise training on reactivity and contractility of arterial strips from hypertensive rats. AB - To determine whether the lowered resting blood pressure values in hypertensive rats were associated with changes in vascular reactivity and/or contractility, helical strips were obtained from different arteries from exercise-trained (running and swimming) hypertensive rats and from running normotensive rats. Each subgroup contained nontrained controls for comparison. Changes in muscle aerobic enzymes, maximum O2 consumption, and body weight indicated that a training effect had occurred. When norepinephrine was added in a dose-response manner to the testing chamber containing helical strips from either the descending aorta, femoral artery, or renal artery, there were no significant differences in reactivity (the negative log dose-response curve) attributable to the training of hypertensive or normotensive groups. However, the trained hypertensive rats exhibited a trend for lower contractility values (dyn/mm2) in some but not all of the strips tested. From these results, we concluded that the lowered resting blood pressures associated with exercise training of hypertensive rats could not be explained by changes in vascular reactivity of their arterial strips. PMID- 2987179 TI - Selection for mutants altered in the expression or export of outer membrane porin OmpF. AB - Strains in which the lacZ gene (which specifies beta-galactosidase) is fused to a gene encoding an envelope protein often exhibit a phenotype termed overproduction lethality. In such strains, high-level synthesis of the cognate hybrid protein interferes with the process of protein export, and this leads ultimately to cell death. A variation of this phenomenon has been discovered with lacZ fusions to the gene specifying the major outer membrane porin protein OmpF. In this case, we find that lambda transducing phage carrying an ompF-lacZ fusion will not grow on a host strain that constitutively overexpresses ompF. We have exploited this observation to develop a selection for ompF mutants. Using this protocol, we have isolated mutants altered in ompF expression and have identified mutations that block OmpF export. Our results suggest that it should be possible to adapt this selection for use with other genes specifying exported proteins. PMID- 2987181 TI - Structural and functional analyses of the fosfomycin resistance transposon Tn2921. AB - The fosfomycin resistance transposon Tn2921 is flanked by directly repeated sequences homologous to the Tn10-related insertion sequence IS10. The nonrepeated DNA sequences of Tn2921 can be deleted without affecting the transposition ability of the element, showing that at least one of the direct repeats is an active insertion sequence. Transposition of Tn2921 seems to occur through direct transposition, since cointegrates have not been observed. The evolutionary relatedness of Tn2921 and IS10 is discussed. PMID- 2987180 TI - Mutations that define the promoter of ompF, a gene specifying a major outer membrane porin protein. AB - Expression of the major porin structural genes, ompF and ompC, is influenced by medium osmotic strength and requires the products of two regulatory genes, ompR and envZ. To help define the sites required for the expression of both porin genes, we have used a novel selection to identify mutations that decrease ompF transcription. From our assignment of the mRNA start site by the primer extension method, these mutations appear to delineate poorly conserved -10 and -35 consensus promoter regions. In addition, one mutation provides the first genetic evidence that an A residue at position -45 may be important for RNA polymerase recognition. PMID- 2987182 TI - Method for gene replacement in Pseudomonas aeruginosa used in construction of recA mutants: recA-independent instability of alginate production. AB - The availability of a technique for site-directed mutagenesis by gene replacement provides a powerful tool for genetic analysis in any bacterial species. We report here a general technique for gene replacement in Pseudomonas aeruginosa. Genes on fragments of cloned P. aeruginosa DNA, altered by transposon mutagenesis, can be transduced into a recipient strain and can replace homologous genes in the P. aeruginosa genome. In this study we applied this technique to the construction of recA mutants of P. aeruginosa. A cloned segment of P. aeruginosa FRD1 DNA was isolated which encoded a protein analogous to the recA gene product of Escherichia coli. The P. aeruginosa recA gene was able to complement several defects associated with recA mutation in E. coli. Transposon Tn1 and Tn501 insertions in the cloned recA gene of P. aeruginosa were used to generate chromosomal recA mutants by gene replacement. These recA strains of P. aeruginosa were more sensitive to UV irradiation and methyl methane sulfonate and showed reduced recombination proficiency compared with the wild type. Also examined was the effect of recA mutations on the expression of alginate, a virulence trait. Alginate is a capsulelike polysaccharide associated with certain pulmonary infections, and its expression is typically unstable. The genetic mechanism responsible for the instability of alginate biosynthesis was shown to be recA independent. PMID- 2987183 TI - Transposable lambda placMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. AB - We have constructed several derivatives of bacteriophage lambda that translocate by using the transposition machinery of phage Mu (lambda placMu phages). Each phage carries the c end of Mu, containing the Mu cIts62, ner (cII), and A genes, and the terminal sequences from the Mu S end (beta end). These sequences contain the Mu attachment sites, and their orientation allows the lambda genome to be inserted into other chromosomes, resulting in a lambda prophage flanked by the Mu c and S sequences. These phages provide a means to isolate cells containing fusions of the lac operon to other genes in vivo in a single step. In lambda placMu50, the lacZ and lacY genes, lacking a promoter, were located adjacent to the Mu S sequence. Insertion of lambda placMu50 into a gene in the proper orientation created an operon fusion in which lacZ and lacY were expressed from the promoter of the target gene. We also introduced a gene, kan, which confers kanamycin resistance, into lambda placMu50 and lambda placMu1, an analogous phage for constructing lacZ protein fusions (Bremer et al., J. Bacteriol. 158:1084 1093, 1984). The kan gene, located between the cIII and ssb genes of lambda, permitted cells containing insertions of these phages to be selected independently of their Lac phenotype. PMID- 2987184 TI - Genetic analysis of mutations that compensate for loss of Escherichia coli DNA topoisomerase I. AB - A transposon Tn10 insertion in topA, the structural gene of Escherichia coli DNA topoisomerase I, behaves as an excluded marker in genetic crosses with many strains of E. coli. However, derivative strains that accept this mutant topA allele are readily selected. We show that many of these topA mutant strains contain additional mutations that compensate for the loss of DNA topoisomerase I. Genetic methods for mapping and manipulating such compensatory mutations are described. These methods include a plate-mating test for the ability of strains to accept a topA::Tn10 allele and a powerful indirect selection for transferring compensatory mutations from male strains into non-compensatory female strains. One collection of spontaneous compensatory mutants is analyzed in detail and is shown to include compensatory mutations at three distinct loci: gyrA and gyrB, the genes that encode the subunits of DNA gyrase, and a previously unidentified locus near tolC. Mutations at this third locus, referred to as toc (topoisomerase one compensatory) mutations, do not behave as point mutations in transductional crosses and do not result in lowered DNA gyrase activity. These results show that wild-type strains of E. coli require DNA topoisomerase I, and at least one class of compensatory mutations can relieve this requirement by a mechanism other than reduction of DNA gyrase activity. PMID- 2987186 TI - Streptococcus faecalis sex pheromone (cAM373) also produced by Staphylococcus aureus and identification of a conjugative transposon (Tn918). AB - Streptococcus faecalis RC73 was found to harbor a conjugative plasmid (pAM373) which confers a mating response to a sex pheromone (cAM373) excreted by plasmid free members of the same species. The pheromone was also detected in culture filtrates of all of 23 Staphylococcus aureus strains but in only 2 of 22 coagulase negative staphylococcus strains. Streptococcus sanguis Challis and G9B also produced the activity, but 10 other Streptococcus sanguis strains did not. The activity was also produced by Streptococcus faecium 9790. A tetracycline resistance (Tc) determinant present in S. faecalis RC73 was not associated with pAM373 but served as a useful marker in efforts to identify pAM373 among other plasmids present in the strain. Analyses of the Tc determinant showed that it was located on a conjugative transposon very similar to Tn916. Designated Tn918, the transposon could insert into pAM373 as well as into two other hemolysin plasmids. Whereas pAM373 derivatives transferred very well between strains of Streptococcus faecalis, the plasmid would not establish in Staphylococcus aureus or Streptococcus sanguis. However, a derivative of pAM373 carrying Tn918 proved to be a useful delivery vehicle for generating transposon insertions into multiple sites on the staphylococcal chromosome. PMID- 2987185 TI - Cloning and expression in Escherichia coli of sdhA, the structural gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex. AB - Bacillus subtilis cytochrome b558 is a transmembrane protein which anchors succinate dehydrogenase (SDH) to the cytoplasmic membrane and is reduced by succinate. The structural gene for this cytochrome was cloned and expressed in Escherichia coli. Random BamHI or BglII fragments of B. subtilis 168 DNA were cloned in the BamHI site of plasmid pHV32. The derived plasmids were used to transform B. subtilis SDH mutants to chloramphenicol resistance by integration of the plasmid via DNA homology. Of some 3,000 transformants tested, 6 were SDH positive and had pHV32 integrated close to the sdh operon. Two plasmids, pKIM2 and pKIM4, with an insert of B. subtilis DNA of 5.7 and 3.4 kilobases, respectively, were generated by transforming E. coli with DNA from the SDH positive transformants after cleavage with EcoRI or BglII and ligation. In E. coli carrying either of the two plasmids, about 4% of total membrane protein was B. subtilis cytochrome b558. E. coli (pKIM2) also contained antigen which reacted with antibodies specific for the flavoprotein and the iron-sulfur protein subunit of B. subtilis SDH. Enzymatically active, membrane-bound B. subtilis SDH could not be demonstrated in E. coli (pKIM2). The B. subtilis DNA insert in pKIM2 could transform B. subtilis sdhA (cytochrome b558), sdhB (flavoprotein), and sdhC (iron sulfur protein) mutants to the wild type. The results suggest that pKIM2 carries the whole B. subtilis sdh operon. The data confirm the gene order and the proposed direction of transcription of the B. subtilis sdh operon. Most likely the sdh genes in E. coli(pKIM2) are controlled by their natural promoter. PMID- 2987187 TI - Mapping of transfer regions within incompatibility group HI plasmid R27. AB - Plasmids of incompatibility group HI are large (greater than 150 kilobases [kb]) and possess an unusual thermosensitive mode of conjugative transfer. R27, the prototype IncHi1 plasmid, encodes resistance to tetracycline via a determinant which is related to transposon Tn10. A restriction endonuclease map of R27 (size, 182 kb) was recently constructed with ApaI, PstI, and XbaI. Transfer genes within R27 were mapped by insertion of Tn5 and Tn7. At least two different regions of the plasmid were concerned with transfer functions. Insertions into either region completely abolished transfer. None of the insertions had any effect on entry exclusion (Eex) of other IncH plasmids. However, a deletion mutant which lacked the Eex function was obtained, allowing us to map the probable site of the gene encoding Eex to one of the two transfer regions. The tetracycline resistance determinant in R27 was located within an 8-kb region between the two main transfer regions. The transfer genes, therefore, are not located together in R27 but are situated in at least two major widely separated transfer regions. PMID- 2987189 TI - Restriction endonuclease analysis of the ilvGEDA operon of members of the family Enterobacteriaceae. AB - Four of the genes required for the biosynthesis of isoleucine and valine form the ilvGEDA operon in Escherichia coli K-12 and Salmonella typhimurium. The structural relationship of these genes was examined in eight other members of the family Enterobacteriaceae by genomic Southern blot hybridization. These genes are contiguous in all the strains examined, and specific restriction sites appear to be highly conserved, indicating the possible functional importance of the DNA sequences of which they are part. PMID- 2987188 TI - Organization of genes necessary for growth of the hydrogen-methanol autotroph Xanthobacter sp. strain H4-14 on hydrogen and carbon dioxide. AB - Mutants unable to grow on H2 and CO2 were isolated in the hydrogen-methanol autotroph Xanthobacter sp. strain H4-14 and complemented with a clone bank constructed in a broad-host-range cosmid vector. The mutants fell into two classes. Class I mutants (Cfx-) cannot grow on hydrogen or methanol and are deficient in one or more of the key enzymes of the Calvin Cycle. Class II mutants (Hox-) can grow on methanol but not on hydrogen and lack hydrogenase activity. Restriction maps of the complementing clones show that each class is not linked to the other. Subcloning and Tn5 mutagenesis have localized the regions of DNA complementing these mutants. The region complementing a class I mutant which is deficient in ribulosebisphosphate carboxylase activity is approximately 3.2 kilobase pairs in size. Expression of this enzyme activity from cloned DNA gave evidence for the orientation of an operon containing the structural genes for this enzyme. The region complementing most of the class II mutants is 3 to 4.5 kilobase pairs in size. PMID- 2987190 TI - SAL-TOL in vivo recombinant plasmid pKF439. AB - SAL-TOL in vivo recombinant plasmid pKF439 was characterized in a strain from a mixed culture of bacteria harboring various degradative plasmids. Analysis of the gene organization of pKF439 revealed that the 57-kilobase TOL fragment, including the 40-kilobase TOL metabolic region, was inserted into the complete SAL replicon at the position of SmaI-C within XhoI-B of SAL. The molecular size of pKF439 was calculated to be 138 kilobases. pKF439 could be transferred to Pseudomonas putida and Pseudomonas aeruginosa at high frequency, and the transconjugants gained the ability to grow with m-xylene, m-toluate, and salicylate as the sole carbon source. PMID- 2987191 TI - Broad-host-range plasmid pRK340 delivers Tn5 into the Legionella pneumophila chromosome. AB - Transposon Tn5 was introduced into Legionella pneumophila on plasmid pRK340, which is temperature sensitive for plasmid maintenance. The presence of plasmid DNA was confirmed by agarose gel electrophoresis and by conjugal transfer of the plasmid to Escherichia coli. Tn5 insertions were obtained by culturing L. pneumophila at the nonpermissive temperature (43 degrees C) on buffered charcoal yeast extract agar containing kanamycin. Of the 260 kanamycin-resistant colonies picked, 220 failed to conjugate pRK340 to E. coli. Plasmid DNA was not visualized from eight randomly picked Tn5-containing strains, and Southern hybridizations indicated that Tn5, but not pRK340, inserted into multiple sites in the Legionella chromosome. In addition, the streptomycin resistance determinant on Tn5 was expressed in L. pneumophila. PMID- 2987192 TI - Transport of vitamin B12 in Escherichia coli: cloning of the btuCD region. AB - The transport of vitamin B12 in Escherichia coli requires a specific vitamin B12 receptor protein in the outer membrane and the tonB gene product. In addition, the btuC gene, located at min 38 on the genetic map, has been found to influence vitamin B12 uptake or utilization. The btuC function is required for the growth response to vitamin B12 when the outer membrane transport process (btuB or tonB function) is defective. However, even in a wild-type strain, btuC is required for proper transport of vitamin B12. Additional mutations in the vicinity of btuC were isolated as lac fusions that produced a phenotype similar to that of a btuC mutant. The btuC region was cloned by selection for complementation of a btuC mutation. Complementation testing with plasmids carrying various deletions or transposon Tn1000 insertions demonstrated that the new mutations defined a separate, independently expressed locus, termed btuD. The coding regions for both genes were identified on a 3.4-kilobase HindIII-HincII fragment and were 800 to 1,000 base pairs in length. They were separated by a 600- to 800-base-pair region. The gene order in this portion of the chromosome map was found to be pps zdh-3::Tn10-btuD-btuC-pheS. Expression of beta-galactosidase in the btuD-lac fusion-bearing strains, whether proficient or defective in vitamin B12 transport, was not regulated by the presence of vitamin B12 in the growth medium. PMID- 2987193 TI - Kinetic studies on formation of cytochrome oxidase of Rhodopseudomonas capsulata after a shift from phototrophic to chemotrophic growth. AB - Rhodopseudomonas capsulata cells were shifted from phototrophic (anaerobic, light) to chemotrophic (semiaerobic, dark, 10% air saturation) growth conditions. During the adaptation period of 4 h, the bacteriochlorophyll content of cells and membranes decreased, and a newly synthesized 65-kilodalton polypeptide of the cytochrome oxidase was incorporated into the membrane fraction. The enzymatic activity of the cytochrome oxidase increased strongly after a lag time of 2 h. The amount of cytochrome oxidase protein does not follow the same kinetics. The relative amount of a membrane-bound cytochrome c of low molecular weight, which has been proposed to be a donor for the cytochrome oxidase, increased during adaptation. PMID- 2987194 TI - Transmembrane permeability channels across the outer membrane of Haemophilus influenzae type b. AB - Outer membranes of Haemophilus influenzae type b were fractionated to yield Triton X-100-insoluble material and lipopolysaccharide and phospholipids. Liposomes reconstituted from lipopolysaccharide and phospholipids were impermeable to sucrose (Mr, 342) and to a high-molecular-weight dextran (average Mr, 6,600). When the Triton X-100-insoluble material was introduced into the reconstituted liposomes, the vesicles became permeable to sucrose, raffinose (Mr, 504), and stachyose (Mr, 666) and fully retained dextrans of Mr greater than 1,500. Inulin (average Mr, 1,400) was tested for its efflux from the reconstituted outer membrane vesicles; 62% of the added inulin was trapped. The molecular weight exclusion limit for the outer membrane of H. influenzae type b was therefore estimated at approximately 1,400. A protein responsible for the transmembrane diffusion of solutes was purified from H. influenzae type b by extraction of whole cells with cetyl trimethyl ammonium bromide. When this extract was passed over DEAE-Sepharose, three protein-containing peaks (I, II, and III) were eluted. Peaks I and II contained mixtures of proteins as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; when tested for their pore-forming properties, these proteins were unable to render liposomes of lipopolysaccharide and phospholipid permeable to sucrose. Peak III contained only one molecular species of protein of molecular weight 40,000; this protein acted as a porin in reconstituted vesicles. The molecular weight exclusion limit for 40,000-molecular-weight protein matched the estimate of approximately 1,400 which was determined for outer membranes. A series of homologous saccharides of increasing degree of polymerization was prepared from agarose by hydrolysis with beta-agarase and fractionation on gel filtration chromatography. These oligosaccharides of Mr, 936, 1,242, 1,548, and 1,854 were assayed for retention by the complete vesicles containing 40-kilodalton protein and lipopolysaccharide and phospholipids. All of these oligosaccharides were lost by efflux through the porin. Since the molecular conformation of the largest oligosaccharide is an elongated semirigid helix, it is suggested that the pore formed by the 40 kilodalton protein does not act as a barrier to the diffusion of this compound. PMID- 2987195 TI - Alteration in cation specificity of the melibiose transport carrier of Escherichia coli due to replacement of proline 122 with serine. AB - The structural genes (melB) for the melibiose carrier of five mutants of Escherichia coli showing altered cation specificity for melibiose transport were cloned. The mutations were mapped in a 248-base-pair DNA fragment by a recombinational assay by using the mutants transformed with hybrid plasmids carrying various portions of the wild-type melB gene. The nucleotide sequences of the corresponding DNA fragments derived from mutated melB genes were determined, and the amino acid sequences of the carrier were deduced. Proline 122 was replaced with serine in the melibiose carrier of all five mutants (which were isolated independently). We conclude that this amino acid replacement caused the alteration in cation specificity (loss of coupling to H+) of the melibiose carrier. PMID- 2987196 TI - Analysis of the active center of hydrogenase from Desulfovibrio vulgaris Miyazaki by magnetic measurements. AB - Hydrogenase [hydrogen: ferricytochrome c3 oxidoreductase, EC 1.12.2.1] solubilized and purified from the particulate fraction of Desulfovibrio vulgaris Miyazaki F (IAM 12604) contains 8 iron and 8 labile sulfide ions in one molecule which is composed of two unequal subunits (Mr: 60,000 + 29,000). It does not contain nickel atoms. The EPR (electron paramagnetic resonance) spectrum has an isotropic signal at g = 2.017 which is independent of the temperature. The peak to-peak width of the signal is about 20 G. The signal intensity is nearly equivalent to 1 unpaired electron per molecule. No other signals can be detected in the field range between 2,240 and 4,240 G (which corresponds to g-values between 2.91 and 1.54). Ferricyanide has only a little effect on the shape and intensity of the EPR signal. The hydrogenase reduced under H2 is EPR silent. The Mossbauer spectrum has no hyperfine splitting at 4K. The isomer shift and quadrupole splitting at 77K are 0.38 and 0.87 mm/s, respectively. Based on these magnetic measurements, the structure of the active center of hydrogenase was suggested to be [4Fe-4S]3+ + [4Fe-4S]2+. PMID- 2987197 TI - Purification and characterization of S-adenosylhomocysteine hydrolase from mouse mastocytoma P-815 cells. Evidence for cell cycle-specific fluctuation of the enzyme activity. AB - S-Adenosylhomocysteine hydrolase [EC 3.3.1.1] was purified to electrophoretic homogeneity from mastocytoma P-815 cells. The purified enzyme had a molecular weight of 190,000, as estimated by Sephadex G-200 chromatography, and a monomer molecular weight of 45,000, as determined by polyacrylamide gel electrophoresis in the presence of SDS. The Km value for adenosine was 0.29 microM and the Vmax value 4.5 mumol S-adenosylhomocysteine X min-1 X mg-1 in the synthetic reaction, while the Km value for S-adenosylhomocysteine was 0.77 microM and the Vmax 0.48 mumol adenosine X min-1 X mg-1 in the hydrolytic reaction. The purified enzyme also had one binding site for adenosine (KD = 2.61 X 10(-7) M) and one for cAMP (KD = 1.6 X 10(-7) M). Using rabbit antiserum raised against the purified enzyme, it was shown that the enzyme activity and enzyme synthesis fluctuated during the cell cycle of mastocytoma cells, reaching the maximum levels as the cells changed from the G1/S phase to the G2 phase. PMID- 2987198 TI - Heme-linked spectral changes of the protein moiety of hemoproteins in the near ultraviolet region. AB - Spectral changes of hemoproteins in the near ultraviolet region on binding to a ligand and on oxidation-reduction of the heme-iron were studied by computer controlled spectrophotometry. Near ultraviolet difference spectra between the low spin and high spin forms of ferric hemoproteins were classified into three groups: Those showing two absorption peaks having maxima at around 285 and 295 nm, those showing a peak at around 275 nm, and those showing a peak at around 300 nm. No corresponding absorption peak was observed with model heme complexes of low molecular weight. The intensity of the peak in cyanide difference spectra of catalase and horseradish peroxidase in the near ultraviolet region was dependent on the concentration of added cyanide and paralleled the intensity of the spectral changes in the Soret region. The spectral changes in both the near ultraviolet and Soret regions developed within 6 ms after the addition of cyanide. Difference spectra between the reduced and oxidized forms of cytochrome c, cytochrome oxidase-cyanide complex, hemoglobin, and lactoperoxidase-cyanide complex showed a characteristic peak at around 285-290 nm. Various difference spectra of hemoglobin in the near ultraviolet region were also measured. The observed positions, shapes, combinations, and relative intensities of the peaks were compared with those of solvent perturbation difference spectra and pH difference spectra of proteins and aromatic amino acids and also with the diacetylchitobiose-induced difference spectrum of lysozyme. The kinds of aromatic amino acid residues possibly responsible for the observed difference peaks were discussed on the basis of the results of the comparison. Based on the results obtained, the common occurrence of a heme-linked functional response of the hemoprotein conformation was suggested. PMID- 2987199 TI - Introduction of mouse C epsilon genes into Cos-7 cells and fertilized mouse eggs. AB - A fragment of the cloned gene for the mouse C epsilon chain, coding for the first, second, third, and fourth domains, has been coupled to the SV40 promotor region (pSV2-mC epsilon). About 50 copies of pSV2-mC epsilon or its PvuII-EcoRI fragments were introduced into Cos-7 cells. Expression of PvuII-EcoRI fragments of pSV2-mC epsilon was observed in about 50% of the Cos-7 cells by indirect fluorescence staining. However, no expression of circular pSV2-mC epsilon was observed. About 200 copies of linearized pSV2-mC epsilon with EcoRI were introduced into fertilized mouse eggs. Two of 78 mice born from these eggs had integrated mouse C epsilon genes. Mouse C epsilon gene was shown to be integrated in a tandem array and as intact structures without undergoing gross deletions or rearrangements, judged from the Southern blotting patterns from several restriction enzymes. The first transgenic mouse was mated to a normal male to examine whether mouse C epsilon gene were stably transmitted to progeny. Among 5 mice to which the C epsilon gene had been transmitted, one deleted 5 copies of this gene and another deleted one junction fragment, thus demonstrating relatively unstable transmission. No C epsilon mRNA was detected in the liver, kidney, brain, lung, skeletal muscle, heart, testis, or spleen of a transgenic mouse. PMID- 2987200 TI - Autocatalytic activation of C1r subcomponent of the first component of human complement. AB - Autoactivation of the proenzyme form of a subunit of the first component (C1r) was performed in the presence and absence of diisopropyl fluorophosphate (DFP). The time-course of autoactivation of zymogen C1r followed a sigmoidal curve and was accelerated by addition of the enzyme C1r and by increasing the concentration of C1r, suggesting that autoactivation of C1r consists of two intermolecular reactions, i.e. zymogen(C1r)- and enzyme(C1r)-catalyzed reactions. In the presence of 10 mM DFP, the enzyme-catalyzed autoactivation of C1r was completely inhibited, while the zymogen-catalyzed autoactivation still proceeded depending upon C1r concentration. These results suggested that the zymogen-catalyzed autoactivation of C1r is a DFP-insensitive second-order reaction and is mediated by an active site generated in a single chain C1r through a conformational change (Kassahara et al. (1982) FEBS lett. 141, 128-131). Based on these results, a possible reaction process of autoactivation of C1r was proposed, as follows: (formula; see text) where C1r represents a conformational isomer which catalyzes the autoactivation of C1r, and the rate constants, k2 and k3, are of second order. Utilizing a computer, we simulated the autoactivation of C1r and found the above scheme to be a reasonable model of C1r autoactivation. Evidence which supports the formation of a conformational isomer of C1r, C1r, as an intermediate in its autoactivation was also obtained by a surface radiolabeling method. PMID- 2987201 TI - Induction of spermidine/spermine N1-acetyltransferase by parathyroid hormone in rabbit costal chondrocytes in culture. AB - Parathyroid hormone (PTH) increased the activity of spermidine/spermine N1 acetyltransferase, a rate-limiting enzyme of polyamine biodegradation, in rabbit costal chondrocytes in culture. The enzyme activity increased in a dose-dependent manner after addition of PTH to the culture, reaching a maximum at 8 h. The increase in the enzyme activity was abolished by cycloheximide or actinomycin D. Dibutyryl cyclic AMP also induced the acetyltransferase to some extent. These results suggest that the induction of spermidine/spermine N1-acetyltransferase by PTH may play some significant role in the expression of the differentiated phenotype of chondrocytes. PMID- 2987202 TI - Phosphatidate phosphatase in rat liver: the relationship between the activities with membrane-bound phosphatidate and aqueous dispersion of phosphatidate. AB - Gel filtration of rat liver cytosol on Bio-Gel A-5m resolved the phosphatase activities into four peaks, all of which showed activity with either phosphatidase bound to microsomal membrane (PAmb) or phosphatidate dispersed in sonicated microsomal lipid (PAaq) as the substrate. A major part of the PAmb phosphatase activity (52%) was eluted in a peak with an apparent molecular weight (Mr) of 500,000 where the PAaq phosphatase activity was very low. A major PAaq phosphatase activity peak (48%) was obtained in the void volume (Vo), where the PAaq phosphatase activity was higher than the PAmb phosphatase activity. The addition of 0.075% Tween 20 to the elution buffer gave only the 500 kilodalton (kDa) peak. When the activity in the Vo peak obtained in the absence of Tween 20 was rechromatographed in the presence of the detergent, a part of the activity was dissociated into 500 kDa molecules having a preference for PAmb. These results suggest that the enzymes obtained in the Vo peak are formed by the association of the 500 kDa molecules with macromolecules and that the substrate preference of phosphatidate phosphatase is modified by the change in the physical state of the enzyme. The microsomal phosphatidate phosphatase activities were also separated on Bio-Gel A-5m after solubilizing by sonication. Most of both the PAmb and PAaq phosphatase activities were coeluted in the Vo peak, in which the latter activity was higher than the former. When the gel filtration was performed in the presence of Tween 20, a major activity peak with a preference for PAmb was obtained at the elution volume corresponding to apparent Mr 500,000, indicating a potential relationship between the cytosolic and microsomal activities. PMID- 2987203 TI - A low potential iron-sulfur cluster bound to membranes in blue-green algae. AB - A new low temperature EPR signal at g = 1.92 was found in photoreduced membrane fractions prepared from blue-green algae, Nostoc muscorum, Tolypothrix tenuis, Phormidium persicinum, and Anabaena variabilis. The signal also appeared when the sample was chemically reduced in the dark. Judging from the spectrum, the signal was probably due to the reduced form of a membrane-bound iron-sulfur cluster, tentatively designated as Cluster D. The midpoint potential of Cluster D was determined to be -0.27 volts. Possible roles of the cluster in electron transfer systems are also discussed. PMID- 2987205 TI - Nuclear mutants of Saccharomyces cerevisiae with altered subunits 4, 5, and 6 of cytochrome oxidase. AB - A collection of pet mutants of Saccharomyces cerevisiae has been screened for lesions in cytochrome oxidase. Three different complementation groups have been identified to consist of strains with altered forms of subunits 4, 5, or 6 that are known to be encoded by nuclear genes. The mutant proteins cross-react with antiserum to the holoenzyme or to the individual subunits but exhibit either an increase or decrease in size. In each instance the mutation imparts a respiratory deficient phenotype which is due to reduced levels of cytochrome oxidase activity in the mitochondria. These results indicate that each of the three proteins is required either for the catalytic activity or for the assembly of functional cytochrome oxidase. PMID- 2987204 TI - A clarification of the effects of DCCD on the electron transfer and antimycin binding of the mitochondrial bc1 complex. AB - We have studied in detail the effects of dicyclohexylcarbodiimide (DCCD) on the redox activity of the mitochondrial bc1 complex, and on the binding of its most specific inhibitor antimycin. An inhibitory action of the reagent has been found only at high concentration of the diimide and/or at prolonged times of incubation. Under these conditions, DCCD also displaced antimycin from its specific binding site in the bc1 complex, but did not apparently change the antimycin sensitivity of the ubiquinol-cytochrome c reductase activity. On the other hand, using lower DCCD concentrations and/or short times of incubation, i.e., conditions which usually lead to the specific inhibition of the proton translocating activity of the bc1 complex, no inhibitory effect of DCCD could be detected in the ubiquinol-cytochrome c reductase activity. However, a clear stimulation of the rate of cytochrome b reduction in parallel to an inhibition of cytochrome b oxidation has been found under these conditions. On the basis of the present work and of previous reports in the literature about the effects of DCCD on the bc1 complex, we propose a clarification of the various effects of the reagent depending on the experimental conditions employed. PMID- 2987206 TI - Chemoattractant receptor-induced hydrolysis of phosphatidylinositol 4,5 bisphosphate in human polymorphonuclear leukocyte membranes. Requirement for a guanine nucleotide regulatory protein. AB - Incubation of plasma membranes from human polymorphonuclear leukocytes (PMNs) with [gamma-32P]ATP in the presence of MgCl2 resulted in the formation of 32P labeled phosphatidic acid (PA), phosphatidylinositol 4-phosphate (PIP), and phosphatidylinositol 4,5-bisphosphate (PIP2). Membranes from PMN specific and azurophil granules synthesized only PIP, suggesting that PIP2 metabolism is confined to the plasma membrane in PMNs. Further incubations of the labeled plasma membranes for 60 s in the presence of 1 mM CaCl2 resulted in the hydrolysis of approximately 40 and 50% of the labeled PIP and PIP2, respectively. In the presence of 2 microM added CaCl2, PIP and PIP2 levels were unchanged by incubation with either the chemoattractant N-formyl-methionyl-leucyl phenylalanine (fMet-Leu-Phe) at 0.1 microM or by 10 microM GTP; however, addition of fMet-Leu-Phe plus GTP together resulted in a 11 and 28% decrease in PIP and PIP2, respectively. These treatments had no effect on PA levels. No additional radiolabeled organic-soluble products were detected after treatment with fMet-Leu Phe plus GTP. Incubation of intact PMNs, with the Bordetella pertussis toxin (islet-activating protein) eliminated the ability of fMet-Leu-Phe plus GTP to promote PIP2 breakdown in the isolated plasma membranes, but did not inhibit PIP2 degradation in the presence of 1 mM CaCl2. These results provide the first direct evidence that the fMet-Leu-Phe receptor in PMN membranes is coupled to polyphosphoinositide hydrolysis through an islet-activating protein-sensitive guanine nucleotide regulatory protein. PMID- 2987207 TI - Activation of myocardial neutral triglyceride lipase and neutral cholesterol esterase by cAMP-dependent protein kinase. AB - Lipolysis of intracellular triglycerides in the heart has been shown to be regulated by hormones. However, activation of myocardial triglyceride lipase in a cell-free system has not been directly demonstrated. In the present studies, initial attempts to demonstrate cAMP-dependent activation of triglyceride lipase using the 1,000 X g supernatant fraction (S1) of mouse heart homogenate were unsuccessful, presumably due to the masking effects of high levels of lipoprotein lipase activity even when assayed at pH 7.4 and in the absence of apolipoprotein C-II. Myocardial lipoprotein lipase in the 40,000 X g supernatant fraction was then removed by heparin-Sepharose affinity chromatography. The lipoprotein lipase free fractions were shown to contain neutral triglyceride lipase and neutral cholesterol esterase of about equal activities. The triglyceride lipase and cholesterol esterase activities fell progressively during preincubation in the presence of 5 mM Mg2+. Additions of cAMP and ATP resulted in 40-70% activation of both triglyceride lipase and cholesterol esterase. The activation was blocked by protein kinase inhibitor and was restored by the addition of exogenous cAMP dependent protein kinase. Since lipoprotein lipase has no activity toward cholesteryl oleate, activation of cholesterol esterase in untreated S1 was readily demonstrable. Both triglyceride lipase and cholesterol esterase activities were present in homogenates prepared from isolated rat heart myocytes. We conclude that the myocardium contains a hormone-sensitive lipase that is regulated in a fashion similar to that of the adipose tissue enzyme. PMID- 2987208 TI - Regulation of specific rat liver messenger ribonucleic acids by triiodothyronine. AB - A plasmid cDNA library was constructed using poly(A+) RNA isolated from the livers of rats treated with 3,5,3'-triiodothyronine (T3) and fed a high carbohydrate diet. This library was screened by differential colony hybridization with [32P]cDNA probes made from hypothyroid and hyperthyroid rat liver poly(A+) RNA to obtain clones representing T3-inducible mRNAs. Using plasmid cDNAs to 4 different T3-inducible mRNAs, we have studied by hybridization assay the responses of these mRNAs to different thyroidal steady states and to a high carbohydrate diet. The fold of induction (hypothyroid to hyperthyroid) varied from about 4.0 (mRNA 5-8D) to 13.2 (mRNA 4-12B). The linearity of response with regard to nuclear receptor occupancy was estimated by assessing the relative mRNA levels in a euthyroid state. Three of the mRNAs demonstrated nonlinear responses with the largest portion of the induction occurring in the euthyroid to hyperthyroid transition. An induction by the high carbohydrate diet was clearly seen for only one mRNA (5-8D) suggesting that these two pathways of induction are independent. In a study of the response kinetics of each mRNA to a nuclear receptor saturating dose of T3 in hypothyroid animals, an increase was seen within 4 h (the earliest time point examined) for one of the mRNAs. The other 3 mRNAs did not increase significantly until 8 h after the T3 dose. Northern analysis showed a single mRNA corresponding to each of these 4 clones with sizes ranging from about 1375 to 7600 bases. Two mRNAs (5-9E and 4-12B) were shown by hybrid-selected translation to code for proteins of molecular mass of about 27 and 46 kDa, respectively. The availability of several different cDNA probes to T3 responsive liver mRNAs should facilitate future studies on the mechanism of action of this hormone. PMID- 2987209 TI - Alteration in the characters of CDP-choline synthetase and phospholipid-choline exchange enzyme upon choline starvation in Chinese hamster ovary cells. AB - When CHO-K1 cells are cultivated under choline-deficient conditions, the specific activity of CDP-choline synthetase increases and conversely phospholipid-choline exchange enzyme activity decreases, whereas the other three known enzyme activities related to synthesis of phosphatidylcholine remain unchanged. The changes of the former two enzyme activities take place immediately after removal of choline from the medium. The altered activities readily revert to the control levels upon resupplementation of choline to the starved cell culture. The changes upon choline starvation are sensitive to cycloheximide, while the restoration processes are insensitive to the drug. The activity of CDP-choline synthetase in unstarved control cells is found in both the soluble and membrane fractions. The Km value of the enzyme in the soluble fraction for choline phosphate differs from that in the membrane fraction. Asolectin alters the Km value of the former to a value close to that of the latter and raises its Vmax value, whereas it hardly affects the Km and Vmax values of the latter. In choline-starved cells, the activity is exclusively found in the membrane fraction. The change in the subcellular distribution of the activity upon choline starvation is sensitive to cycloheximide. The altered subcellular distribution reverts to the initial status upon resupplementation of choline even in the presence of cycloheximide. The activity of the phospholipid-choline exchange enzyme is exclusively found in the membrane fraction for both starved and control cells. The properties of the enzyme are altered upon choline starvation with respect to the Vmax value for choline and the Km and Vmax values for Ca2+. These altered kinetic parameters are changed by egg yolk phosphatidylcholine so as to be indistinguishable from those in unstarved control cells. We discuss the mechanism of the alterations in the characters of both enzymes in response to choline starvation. PMID- 2987210 TI - Acetylglutamate kinase-acetylglutamyl-phosphate reductase complex of Neurospora crassa. Evidence for two polypeptides. AB - Mutations at the arg-6 locus in Neurospora crassa are divided into two complementation groups (A and B) and a third noncomplementing group. There are many suppressible nonsense mutations among mutants in complementation group B and one in the noncomplementing group; no nonsense mutations exist among mutants in complementation group A (Davis, R. H., and Weiss, R. L. (1983) Mol. Gen. Genet. 192, 46-50). We show here that the mutants are defective in either or both of two enzymes of arginine biosynthesis, acetylglutamate kinase and/or acetylglutamyl phosphate reductase. Mutants in complementation group A lack acetylglutamate kinase, those in complementation group B lack acetylglutamyl-phosphate reductase, and those in the noncomplementing group lack both activities. Mutants in group B also have reduced levels of acetylglutamate kinase. The enzymes from purified mitochondria are readily separable by gel filtration and by Blue A dye affinity chromatography. Acetylglutamate kinase appears to be an octamer with a molecular weight of 400,000, whereas acetylglutamyl-phosphate reductase appears to be a dimer with a molecular weight of 93,000. This suggests that the two activities reside on distinct polypeptides. These results are best accommodated by the following model: the arg-6 locus encodes a single mRNA which is translated into a single polypeptide; the latter is then cleaved post-translationally to yield two physically separable enzymes. PMID- 2987211 TI - Dissociation of protein kinase C activation from phorbol ester-induced maturation of HL-60 leukemia cells. AB - The role of C-kinase in the induction of maturation of HL-60 promyelocytic leukemia cells was examined using two activators of this kinase, 12-O tetradecanoyl phorbol 13-acetate (TPA) and 1-oleoyl-2-acetylglycerol (OAG). At 10(-8) M, a concentration that induced maturation, TPA effectively stimulated C kinase activity in cell-free preparations by increasing the affinity of the enzyme for Ca2+. Similar activation was observed with 20 micrograms/ml of OAG. At these concentrations, addition of either compound to intact cells stimulated the phosphorylation of cellular proteins. Treatment with TPA resulted in an increased phosphorylation of 14 proteins, 9 of which also changed in response to OAG. In addition to the effects on protein phosphorylation, TPA and OAG both affected choline lipid metabolism. TPA at 10(-8) M stimulated the incorporation of [methyl 3H]choline into phosphatidylcholine, sphingomyelin, and lysophosphatidylcholine. OAG at 20 micrograms/ml had quantitatively similar effects on the labeling of the former two lipids, but did not affect incorporation of choline into lysophosphatidylcholine. Despite the similar biochemical effects of TPA and OAG, the diglyceride was unable to induce HL-60 cell maturation as measured by inhibition of cell growth, development of nonspecific esterase activity, phagocytosis, adherence of cells to plastic, and loss of transferrin receptor activity. The lack of effect is not due to metabolism of OAG; maturation could not be induced by treating cells with fresh OAG every 2 h for a period of 12 h. These results suggest a dissociation of the activation of C-kinase and the induction of HL-60 cell maturation by TPA. PMID- 2987212 TI - Regulation of choline kinase activity and phosphatidylcholine biosynthesis by mitogenic growth factors in 3T3 fibroblasts. AB - The regulation of choline kinase activity by fetal bovine serum and the regulation of phosphatidylcholine biosynthesis by choline kinase have been investigated in 3T3 fibroblasts. Treatment of quiescent 3T3 fibroblasts with serum was shown in previous work to increase phosphocholine pool size and phosphatidylcholine biosynthesis. We now report that treatment of 3T3 cells with serum increased intracellular choline kinase activity by 2-3-fold with a concomitant 2-3-fold decrease of intracellular free choline concentrations. Initial rates of choline transport were the same in quiescent and serum-treated cells, whereas choline kinase activity was 2-3-fold higher in serum-treated cells. As a consequence, free choline concentrations were 2-3-fold lower in serum stimulated cells than in control quiescent cells. Phosphocholine turnover rates were increased 2-fold by serum treatment both as a consequence of a serum dependent increase of phosphocholine pools and as a result of a serum-dependent lowering of the phosphocholine half-life. Thus, the overall response of 3T3 cells to serum stimulation included decreased choline pools and increased choline kinase activity, phosphocholine pool size, phosphocholine turnover, and phosphatidylcholine biosynthesis. PMID- 2987213 TI - Spectral characterization of diarylpropane oxygenase, a novel peroxide-dependent, lignin-degrading heme enzyme. AB - Diarylpropane oxygenase, an H2O2-dependent lignin-degrading enzyme from the basidiomycete fungus Phanerochaete chrysosporium, catalyzes the oxygenation of various lignin model compounds with incorporation of a single atom of dioxygen (O2). Diarylpropane oxygenase is also capable of oxidizing some alcohols to aldehydes and/or ketones. This enzyme (Mr = 41,000) contains a single iron protoporphyrin IX prosthetic group. Previous studies revealed that the Soret maximum of the ferrous-CO complex of diarylpropane oxygenase is at approximately 420 nm, as in ferrous-CO myoglobin (Mb), and not like the approximately 450 nm absorption of the CO complex of the ubiquitous heme monooxygenase, cytochrome P 450. This spectral difference between two functionally similar heme enzymes is of interest. To elucidate the structural requirements for heme iron-based oxygenase reactions, we have compared the electronic absorption, EPR, and resonance Raman (RR) spectral properties of diarylpropane oxygenase with those of other heme proteins and enzymes of known axial ligation. The absorption spectra of native (ferric), cyano, and ferrous diarylpropane oxygenase closely resemble those of the analogous myoglobin complexes. The EPR g values of native diarylpropane oxygenase, 5.83 and 1.99, also agree well with those of aquometMb. The RR spectra of ferric diarylpropane oxygenase have their spin- and oxidation-state marker bands at frequencies analogous to those of aquometMb and indicate a high-spin, hexacoordinate ferric iron. The RR spectra of ferrous diarylpropane oxygenase have frequencies analogous to those of deoxy-Mb that suggest a high-spin, pentacoordinate Fe(II) in the reduced form. The RR spectra of both ferric and ferrous diarylpropane oxygenase are less similar to those of horseradish peroxidase, catalase, or cytochrome c peroxidase and are clearly distinct from those of P-450. These observations suggest that the fifth ligand to the heme iron of diarylpropane oxygenase is a neutral histidine and that the iron environment must resemble that of the oxygen transport protein, myoglobin, rather than that of the peroxidases, catalase, or P-450. Given the functional similarity between diarylpropane oxygenase and P-450, this work implies that the mechanism of oxygen insertion for the two systems is different. PMID- 2987215 TI - On the specificity of cytochrome c synthetase in recognition of the amino acid sequence of apocytochrome c. AB - Two forms of yeast cytochrome c synthetases with different specificities were resolved, one (synthetase I), solubilized from mitochondria or the cell debris with Triton X-100, recognizing not horse apocytochrome c but yeast apo-iso-1 cytochrome c as a substrate and the other (synthetase II) still bound with the particulate fraction from mitochondria after treatment with Triton, recognizing both horse and yeast apocytochromes c. The activity with labeled yeast apo-iso-1 cytochrome c as a substrate of cytochrome c synthetase I can be quantitatively inhibited by nonlabeled Candida krusei apocytochrome c and partially by nonlabeled tuna apocytochrome c but not by nonlabeled horse apocytochrome c indicating a specific amino acid sequence being recognized. However, an enzyme similarly solubilized from beef heart mitochondria recognized both horse apocytochrome c and yeast apo-iso-1-cytochrome c for attachment of heme. In view of the fact that the yeast synthetase II and the beef synthetase can both utilize either horse apocytochrome c or yeast apo-iso-1-cytochrome c as substrates, we suggest that these enzymes may also be involved in biosynthesis of cytochrome c1, that is, the ability to attach heme to apocytochrome c and apocytochrome c1 may have been conserved in eucaryotic cells, and that both synthetases may therefore be homologous. PMID- 2987214 TI - Formation of lipid-linked oligosaccharides by MOPC 315 plasmacytoma cells. Decreased synthesis by a nonsecretory variant. AB - MOPC 315 is a BALB/c plasmacytoma which secretes a trinitrophenol-binding IgA lambda 2 paraprotein. We have investigated the incorporation of [3H]mannose into lipid-linked oligosaccharide precursors in wild-type MOPC 315/J and variant nonsecretory 315/P cells. In pulse labeling experiments, no differences could be detected in the ability of the two cell types to incorporate [3H]mannose into lipid-linked oligosaccharides containing 5 or less mannose residues. In contrast, quantitation of the incorporation of [3H]mannose into larger lipid-linked oligosaccharides and proteins revealed a 49 and 40% decrease, respectively, in the 315/P cells compared to wild-type cells. Further characterization of the lipid-linked structures documented a marked decrease in glucosylated oligosaccharides isolated from 315/P cells. When membranes from the two cell lines were analyzed for their ability to transfer [3H]glucose from UDP [3H]glucose to [3H]glucosylphosphoryldolichol, an apparent deficiency was noted in the 315/P preparations. However, if assay conditions were adjusted to include AMP in the reaction mixtures, no differences in the in vitro synthesis of [3H]glucosylphosphoryldolichol or [3H]glucose-labeled oligosaccharide-lipid could be detected. In these reactions AMP was found to prevent hydrolysis of UDP [3H]glucose by inhibiting nucleotide pyrophosphatase (EC 3.6.1.9), the specific activity of which was determined to be more than 100 times greater in variant 315/P compared to wild-type MOPC 315/J cells. This large difference in specific activity was not accompanied by similar differences in the activity of several other enzymes analyzed. A decrease in whole cell UDP-glucose pool size was not detected in 315/P cells. Therefore, if nucleotide pyrophosphatase is important for the control of substrates for glycosylation, it must regulate nucleotide sugar levels at a site other than the cytoplasm of cells, perhaps at the location of synthesis of the larger lipid-linked oligosaccharides. PMID- 2987216 TI - Millisecond kinetics of ATP synthesis driven by externally imposed electrochemical potentials in chloroplasts. AB - We have used rapid mixing and quenching techniques to measure the initial ATP synthesis rates and the duration of the ATP synthetic capacity derived from artificially imposed proton gradients and valinomycin-mediated K+ diffusion potentials in chloroplasts. The initial rate of ATP synthesis driven by a K+ diffusion potential was 10-fold slower than that driven by an acid-base transition of equivalent electrochemical potential. Total yields of ATP resulting from a K+ concentration shift were only slightly affected by the absence of Cl-, indicating that Cl- permeability does not significantly reduce the K+ diffusion potential. The ATP synthetic capacity decayed with a half-life of 0.2 s in the case of a K+ diffusion potential and a half-life of 1.0 s in the case of an acid base shift. In both cases, ATP, added at the time of the pH or [KCl] shift, slowed the decay of the ATP synthesis rates, indicating that the coupling factor controls a channel for proton efflux, as proposed earlier (Portis, A.R., and McCarty, R.E. (1974) J. Biol. Chem. 249, 6250-6254). Because the proton gradient has a longer half-life than the K+ diffusion potential, when combinations of the two are employed to drive ATP synthesis, the proton gradient will make a larger contribution to the initial rate and total yield than that predicted from a strictly linear proportionality of the initial magnitudes of the two gradients. PMID- 2987217 TI - Linear diffusion of restriction endonucleases on DNA. AB - We have investigated the dependence of the rate of cleavage of DNA by EcoRI, HindIII, and BamHI on the chain length of the substrate. In order to keep the influence of flanking sequences and of nonspecific binding identical for all substrates we have carried out all experiments with the same plasmid DNA which had been digested previously with a variety of different restriction enzymes to give a set of substrates of different lengths. Our results show that depending on the buffer conditions long substrates are cleaved faster than small ones. We interpret these findings to mean that under certain conditions a linear diffusion of the enzymes on the DNA is involved in localizing the recognition sites. For EcoRI the mean diffusion length is approximately 1000 base pairs at 1 mM MgC12 which can be shown by diffusion theory to correspond to a linear diffusion coefficient of 5 X 10(-10) cm2 s-1. At 10 mM MgCl2 the linear diffusion of EcoRI is negligible and does not lead to a significant enhancement of the rate of site localization. In the presence of nonsaturating amounts of one of the prokaryotic histone-like protein Hu (NS 2) small and large DNA substrate are cleaved with identical rate by EcoRI indicating that other proteins bound to the DNA constitute a barrier across which linear diffusion cannot take place. We conclude that linear diffusion, albeit detectable under certain conditions in vitro, probably is of little importance for the process of site localization in vivo. PMID- 2987218 TI - The mechanism of insulin stimulation of (Na+,K+)-ATPase transport activity in muscle. AB - Since the mechanism underlying the insulin stimulation of (Na+,K+)-ATPase transport activity observed in multiple tissues has remained undetermined, we have examined (Na+,K+)-ATPase transport activity (ouabain-sensitive 86Rb+ uptake) and Na+/H+ exchange transport (amiloride-sensitive 22Na+ influx) in differentiated BC3H-1 cultured myocytes as a model of insulin action in muscle. The active uptake of 86Rb+ was sensitive to physiological insulin concentrations (1 nM), yielding a maximum increase of 60% without any change in 86Rb+ permeability. In order to determine the mechanism of insulin stimulation of (Na+,K+)-ATPase activity, we demonstrated that insulin also stimulates passive 22Na+ influx by Na+/H+ exchange transport (maximal 200% increase) and an 80% increase in intracellular Na+ concentration with an identical time course and dose-response curve as insulin-stimulated (Na+,K+)-ATPase transport activity. Incubation of the cells with high [Na+] (195 mM) significantly potentiated insulin stimulation of ouabain-inhibitable 86Rb+ uptake. The ionophore monensin, which also promotes passive Na+ entry into BC3H-1 cells, mimics the insulin stimulation of ouabain-inhibitable 86Rb+ uptake. In contrast, incubation with amiloride or low [Na+] (10 mM), both of which inhibit Na+/H+ exchange transport, abolished the insulin stimulation of (Na+,K+)-ATPase transport activity. Furthermore, each of these insulin-stimulated transport activities displayed a similar sensitivity to amiloride. These results indicate that insulin stimulates a large increase in Na+/H+ exchange transport and that the resulting Na+ influx increases the intracellular Na+ concentration, thus activating the internal Na+ transport sites of the (Na+,K+)-ATPase. This Na+ influx is, therefore, the mediator of the insulin-induced stimulation of membrane (Na+,K+)-ATPase transport activity classically observed in muscle. PMID- 2987219 TI - Phosphoprotein phosphatase inhibitor-2. Identification as a species of molecular weight 31,000 in rabbit muscle, liver, and other tissues. AB - Specific antibodies, raised to purified rabbit skeletal muscle inhibitor-2, were used to analyze for the presence of inhibitor-2 in extracts of rabbit skeletal, cardiac, and diaphragm muscles, liver, kidney, brain, and lung. Direct analyses of the extracts by "Western blotting" revealed several immunoreactive species, apparent molecular weights in the range 26,000-136,000, as well as species with the electrophoretic mobility of inhibitor-2, apparent molecular weight 31,000. When supernatants from boiled extracts were similarly analyzed, most of the immunoreactive material was lost and the species corresponding to inhibitor-2 became prominent. Liver and muscle were studied in more detail; immunoprecipitates from either boiled or unboiled extracts were analyzed by Western blotting. The dominant polypeptide now was the species of apparent molecular weight 31,000, corresponding to inhibitor-2. Higher molecular weight species (115,000 in muscle and 136,000 in liver) were also detectable. The amount of inhibitor-2 detected in immunoprecipitates was not greatly different whether unboiled or boiled tissue extracts were used. In addition, extraction of the precipitates by boiling released material that inhibited purified type 1 protein phosphatase. The results suggest that inhibitor-2 is widely distributed in rabbit tissues and is found predominantly as a form of apparent molecular weight 31,000. In particular, the study provides direct demonstration of a species in rabbit liver with similar properties to rabbit muscle inhibitor-2. PMID- 2987220 TI - Isolation of the fructose-1,6-bisphosphatase gene of the yeast Schizosaccharomyces pombe. Evidence for transcriptional regulation. AB - The fructose-1,6-bisphosphatase structural gene (FBP+) of Schizosaccharomyces pombe has been isolated by genetic complementation of a deficient mutant, which is characterized by the inability to grow on a nonfermentable carbon source such as glycerol. Growth on glycerol-containing medium was restored in a S. pombe fructose-1,6-bisphosphatase-deficient mutant (fbp-16) when it was transformed with a plasmid (pAVO4) carrying FBP+. The transformant displayed a 5-fold increase in enzymatic activity when compared to the parental S. pombe strain. Subcloning of DNA fragments from the 8.5-kilobase (kb) insert of pAVO4 defined a 4-kb DNA fragment which contained the functional FBP+ gene and its regulatory region. When this gene was placed under the control of the lac promoter-operator, functional expression in Escherichia coli was obtained, as deduced by complementation of bacterial fructose-1,6-bisphosphatase mutants. The FBP+ gene encodes a 1.9-kb glucose-repressible transcript whose appearance in S. pombe is correlated with fructose-1,6-bisphosphatase derepression in glycerol-containing medium. We suggest that the regulation of the S. pombe FBP+ gene is exerted at the transcriptional level. The S. pombe FBP+ gene gave rise to a 1.9-kb transcript in Saccharomyces cerevisiae, but not to measurable enzymatic activity. PMID- 2987221 TI - Kinetics and concentration dependence of reversible cAMP-induced modification of the surface cAMP receptor in Dictyostelium. AB - We have previously reported that extracellular cAMP induced a reversible shift, from apparent Mr = 40,000 to 43,000, in the electrophoretic mobility of a polypeptide identified by photoaffinity labeling with [32P]8-N3-cAMP as the cAMP receptor of Dictyostelium (Klein, P., Theibert, A., Fontana, D., and Devreotes, P. (1985) J. Biol. Chem. 260, 1757-1764). In this report, we examine the kinetics and concentration dependence of this stimulus-induced receptor modification. Prior to stimulation, 90% of the receptors migrated as the higher mobility form (Mr = 40,000) and 10% as the lower mobility form (Mr = 43,000). Following 15 min of persistent stimulation with 1 microM cAMP, the per cent of receptors migrating as the lower mobility form rose to 80%. This transition occurred with a half-time of 2.5 min. Removal of the stimulus initiated a return to the basal state which occurred with a half-time of about 6 min at 22 degrees C. No reversal occurred at 0 degrees C. Addition and removal of a 50 nM cAMP stimulus induced transitions with similar kinetics, but the final plateau value reached was only 40% lower mobility form. The stimulus concentration which induced 50% of the maximal transition from higher to lower mobility forms at steady state was 27 nM, similar to the KD for [3H]cAMP binding. Scatchard analysis of [3H]cAMP binding indicated that, although a 20% down-regulation occurs during cAMP stimulation, there is no significant difference in the affinities of the higher and lower mobility forms of the receptor. The unoccupied higher and lower mobility forms of the receptor, designated R and D, are considered to be in rapid equilibrium with liganded forms, designated RL and DL. The rate constants for interconversion of the receptor forms R (Formula: see text) D and RL (Formula: see text) DL were calculated from the kinetic data: k1 = 0.012, k-1 = 0.104, k2 = 0.222, and k-2 = 0.055. The interconversion steps are not at equilibrium, suggesting that an energy expenditure occurs during the receptor modification. The pattern of modulation of the cAMP-induced receptor modification suggests that it may be the biochemical mechanism of adaptation. PMID- 2987222 TI - On the mechanism of regulation of type I phosphoprotein phosphatase from bovine heart. Regulation by a novel intracyclic activation-deactivation mechanism via transient phosphorylation of the regulatory subunit by phosphatase-1 kinase (FA). AB - Adenosine 5'-(gamma-thio)triphosphate (ATP gamma S) can substitute for ATP in the activation of the ATP X Mg2+-dependent form of bovine heart type I protein phosphatase (Mr = 75,000) catalyzed by phosphatase-1 kinase (FA). ATP gamma S activates the enzyme to a lower level than ATP, but it phosphorylates the regulatory (R)-subunit to a much higher extent. An [35S]phosphatase-1 [( 35S]E-P) has been isolated, identified, and shown to be a key intermediate in the activation reaction. Treatment of [35S]E-P with dimethyl suberimidate results in cross-linking of the Mr = 34,000 [35S]R-subunit with the Mr = 40,000 catalytic (C)-subunit to form a Mr = 75,000 species, indicating that phosphorylation is not accompanied by dissociation of the holoenzyme. The catalytically active form (Ea) is not the phosphorylated enzyme intermediate. Instead, Ea is directly produced from the intermediate by a Mg2+-dependent, intramolecular autodephosphorylation reaction. The isolated Ea derived from [35S]E-P or from ATP-activated phosphatase 1 has the same half-life (23 min at 30 degrees C). It spontaneously deactivates, via an intramolecular process, to a resting state (Er) which can be fully reactivated by FA X ATP X Mg2+. The deactivation of Ea can be accelerated by chelators, PPi greater than ATP X Mg2+ blocks the PPi effect. Limited trypsinization selectively digests the R-subunit and the resulting C-subunit is Mg2+-dependent. Based on the present data, a novel intracyclic activation deactivation mechanism via transient phosphorylation of the R-subunit is proposed for regulation of phosphatase-1. (formula; see text). PMID- 2987223 TI - The regulatory subunit of neural cAMP-dependent protein kinase II represents a unique gene product. AB - Although the major form of soluble cAMP-dependent protein kinase in bovine cerebral cortex can be classified as a type II kinase, the regulatory subunit (RII) can be distinguished from RII found in other tissues such as heart. Heart and brain RII were distinguished qualitatively by autophosphorylation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The mobility of dephosphorylated heart RII shifted from an apparent Mr of 55,000 to 57,000 following autophosphorylation. In contrast, when RII purified from brain was autophosphorylated with [gamma-32P]ATP, two radiolabeled bands were visualized, a minor band (less than or equal to 20%) which migrated with an Mr of 57,000 similar to the heart protein and a band with Mr = 55,000 which did not shift its mobility in response to autophosphorylation. Brain RII was further distinguished from heart RII on the basis of cAMP binding. Millipore filtration and equilibrium dialysis indicated that 2 mol of cAMP bound/mol of RII in contrast to 4 mol/mol with heart RII. Immunological differences were also apparent. Radioimmunoassays using monoclonal antibodies to RII showed that the brain protein had less than 4% of the cross-reactivity of heart RII. Both immunoblotting and immunoprecipitation using monoclonal as well as serum antibodies established that the cross reactivity in phosphorylated brain RII was associated exclusively with the 57,000 component that behaved like heart RII. The lack of cross-reactivity of neural RII with two different monoclonal antibodies targeted the hinge region of RII as an area where structural differences might be anticipated, and comparative sequence analysis of this region definitively established that the major form of RII in brain is a unique gene product from the RII expressed in heart. PMID- 2987224 TI - Functional analysis of a beta-globin gene containing a TATA box mutation from a Kurdish Jew with beta thalassemia. AB - We recently reported a TATA box mutation (ATAAAA to ATACAA) in a cloned beta globin gene from a Kurdish Jew with homozygous beta thalassemia (Poncz, M., Ballantine, M., Solowiejczyk, D., Barak, I., Schwartz, E., and Surrey, S. (1982) J. Biol. Chem. 257, 5994-5996). We have now introduced this gene into HeLa cells after CaPO4 precipitation of the DNA and studied expression by analyzing globin gene transcripts with a novel S1 nuclease mapping assay. Quantitative and qualitative comparison with the normal beta-globin gene revealed a promoter-down phenotype in the TATA box mutant, with normal RNA processing, and a normal start site for initiation of the primary transcript. Decreased transcriptional efficiency was confirmed directly by analysis of run-off transcripts using assays in vitro. The patient's phenotype of beta thalassemia major is probably the result of two different mutations since haplotype analysis of the beta-like globin gene clusters in genomic DNA from this patient shows heterozygosity for the Mediterranean-type haplotypes I and VII, with the TATA box mutation on a haplotype I chromosomal background. PMID- 2987225 TI - The Mr 28,000 gap junction proteins from rat heart and liver are different but related. AB - The sequence of the amino-terminal 32 residues of the rat heart Mr 28,000 gap junction protein presented here allows, for the first time, a sequence comparison of gap junctional proteins from different tissues (heart and liver). Comparison of the rat heart gap junction protein sequence and that available from rat liver reveals 43% sequence identity and conservative changes at an additional 25% of the positions. Both proteins exhibit a hydrophobic domain which could represent a transmembrane span of the junction. This result unequivocally demonstrates the existence of at least two forms of the gap junction protein. As yet, no homology is evident between the gap junctional proteins of either heart or liver and main intrinsic protein from rat eye lens. PMID- 2987226 TI - Regulation of the oxidative response of human granulocytes to chemoattractants. No evidence for stimulated traffic of redox enzymes between endo and plasma membranes. AB - Experiments were performed to probe the role of exocytotic and endocytotic processes in the regulation of the human granulocyte O-2-generating system. Analytical subcellular fractionation studies indicated that 25-30% of the total cellular b-cytochrome and 8-10% of the flavin co-sedimented with plasma membrane markers, irrespective of stimulation of the cells by the chemoattractants N formyl-Met-Leu-Phe (FMLP) or C5a. Phorbol myristate acetate stimulation resulted in significant translocation of b-cytochrome but not flavin from the specific granule/Golgi to the plasma membrane-enriched fractions. These results indicated that approximately 3.1 X 10(5) flavin and 0.8-1 X 10(6) b-cytochrome molecules are present in the plasma membrane of an isolated unstimulated human granulocyte and that these levels are invariant upon stimulation with chemoattractants. Maximal instantaneous rates of O-2 generation by cells in these preparations, however, were equivalent for all the stimuli. Since stimulation of granulocytes by phorbol myristate acetate, FMLP, or C5a results in exocytosis and/or endocytosis, then the role of these processes in regulating stimulated O-2 production by controlling the content of plasma membrane redox enzymes is questionable. This conclusion was supported by observations made with cytoplasts, which do not have an intracellular reserve of granules. Cytoplasts prepared from granulocytes produced O-2 at equivalent rates as their parent cells on a per unit surface area basis. These results suggest: 1) that stimulation of granulocytes with chemotactic peptides leads to full generation of O-2 at the cell surface without exocytotic recruitment of additional b-cytochrome and flavoprotein from the cytoplasmic compartment; 2) that these redox enzymes are not internalized along with chemoattractant receptors; and 3) that traffic of these redox enzymes between endo- and plasma membranes is not involved in the regulation of O-2 production in suspensions of human granulocytes stimulated by chemoattractants. PMID- 2987227 TI - Wound-induced proteinase inhibitors from tomato leaves. I. The cDNA-deduced primary structure of pre-inhibitor I and its post-translational processing. AB - A cDNA containing the coding region for the complete amino acid sequence of wound induced proteinase Inhibitor I from tomato leaves was constructed in the plasmid pUC9 and characterized. The open reading frame codes for a protein of 111 amino acids. This deduced amino acid sequence revealed the presence of a 42-amino acid N-terminal sequence that is not found in the native protein. This sequence appears to contain a 23-amino acid segment typical of a signal sequence followed by a 19-amino acid sequence containing 9 charged amino acids. The 42-amino acid sequence is apparently lost during maturation to the native Inhibitor I and represents 38% of the translated protein. The Inhibitor I amino acid sequence contains 71% identity with potato tuber Inhibitor I sequence and 35% identity with an inhibitor from the leech. PMID- 2987228 TI - Nucleotide sequence of the haptoglobin and haptoglobin-related gene pair. The haptoglobin-related gene contains a retrovirus-like element. AB - Thirty-three kilobase pairs (kb) of human DNA containing the haptoglobin (Hp) and haptoglobin-related (Hpr) gene pair were cloned, and the nucleotide sequence of 21-kb DNA was determined. The two genes are closely linked, with Hpr being 2.2 kb downstream of Hp. Six hundred nucleotides of DNA occur between the two genes that are not found either 5' to the Hp gene or 3' to the Hpr gene. After the duplication event, the first intron of the Hpr gene acquired a 9-kb insert consisting mainly of a retrovirus-like element with a potential primer-binding site homologous to a mouse isoleucine tRNA. The element forms a repeated family in the human genome that I name RTVL-I (retrovirus-like element-isoleucine). In the coding region of the Hpr gene, there are no frameshift or nonsense mutations and its exon-intron splicing sites, 5' flanking and 3' flanking sequences do not show any obvious defects. There are 28 amino acid differences between the decoded amino acid sequences of the Hpr and Hp genes. Sixteen of these differences occur in the hpr beta chain, and all appear to be located on the surface of the molecule in places not thought to be involved in the hemoglobin binding function of haptoglobin. The structure of the Hpr gene suggests that the gene may be expressed and give rise to a functional product. PMID- 2987229 TI - The A1 adenosine receptor. Solubilization and characterization of a guanine nucleotide-sensitive form of the receptor. AB - Adenosine acting through membrane-bound A1 receptors is capable of inhibiting the enzyme adenylate cyclase. A1 adenosine receptors from rat cerebral cortex have been solubilized in high yield and in an active form with the detergent digitonin. The solubilized receptors bind the agonist radioligand (-)-N6-3-[125I] iodo-4-hydroxyphenylisopropyl)adenosine (HPIA) with the same high affinity, demonstrate the same agonist and antagonist potency series and stereo-specificity as the membrane-bound A1 receptor. In addition to maintaining high affinity agonist binding, soluble A1 receptors' affinity for agonists is still modulated by guanine nucleotides. This result contrasts with other adenylate cyclase coupled receptors (beta 2, alpha 2, D2) wherein high affinity agonist binding is lost subsequent to solubilization. To investigate the molecular basis for this difference, solubilized A1 receptors which were labeled with [125I]HPIA either prior to or subsequent to solubilization, were compared by sucrose density gradient centrifugation. Both labeled species demonstrated exactly the same sedimentation properties and display guanine nucleotide sensitivity. This suggests that the same guanine nucleotide-sensitive receptor complex formed in membranes in stable to solubilization and can form a high affinity agonist complex in soluble preparation. The molecular mechanism responsible for the stable receptor complex in this system compared to the beta 2, alpha 2, and D2 systems remains to be determined. PMID- 2987230 TI - Action of the cardiac alpha 1-adrenergic receptor. Activation of cyclic AMP degradation. AB - Using purified rat ventricular myocytes and membranes prepared from them, we have previously found that alpha 1-adrenergic stimulation causes decreased cyclic AMP accumulation and decreased activation of cyclic AMP-dependent protein kinase. We have now analyzed the mechanism by which alpha 1 stimulation is linked to cyclic AMP metabolism. In an adenylate cyclase assay in which carbachol inhibits the stimulatory effect of norepinephrine, the addition of prazosin (alpha 1 antagonist) has no effect on the response to norepinephrine. In membranes prepared from myocytes treated with pertussis toxin, norepinephrine competes for alpha 1-receptors (assessed by [3H]prazosin binding) with two components, binding to the high affinity component being sensitive to exogenous GTP, exactly as in membranes prepared from control myocytes. In intact cells labeled with [3H]adenine in which carbachol antagonizes the norepinephrine response, prazosin enhances accumulation of [3H]cyclic AMP due to norepinephrine. Treatment of cells with pertussis toxin eliminates inhibition by carbachol but does not alter prazosin's capacity to enhance the norepinephrine response. Addition of phosphodiesterase inhibitors eliminates this effect of alpha 1 blockade. In [3H]adenine-labeled cells loaded with [3H]cyclic AMP by prior treatment with isoproterenol, alpha 1-adrenergic stimulation enhances disappearance of [3H]cyclic AMP. Measurements of cellular cyclic AMP give results similar to those obtained with the adenine labeling technic. We conclude that occupation of the myocyte alpha 1-receptor results in stimulation of cyclic AMP phosphodiesterase activity. PMID- 2987231 TI - Resonance Raman study of cytochrome b562-o complex, a terminal oxidase of Escherichia coli in its ferric, ferrous, and CO-ligated states. AB - Cytochrome b562-o complex, a terminal oxidase in the respiratory chain of aerobically grown Escherichia coli, has been studied by resonance Raman spectroscopy in its air-oxidized, dithionite-reduced, and reduced and CO-ligated states. In the reduced state, with a 406.7-nm excitation, there appeared 1494 and 1473 cm-1 lines, indicating that low spin and high spin components are included in the cytochrome b562-o complex. For the air-oxidized protein, resonance Raman lines were observed at 1372, 1503, and 1580 cm-1 with a 413.1-nm excitation, indicating that there is a ferric low spin heme. In addition, a weak but appreciable Raman line was observed at 1480 cm-1 assignable to a ferric high spin heme. Accordingly, it was concluded that low spin and high spin components are included in the cytochrome b562-o complex in the reduced and the air-oxidized states. In the CO-ligated state, with a defocused laser beam of 413.1 nm, two Raman bands assignable to the Fe-CO stretching mode have been observed at 489 and 523 cm-1, as a major and a minor component, respectively. When the laser beam was focused upon the sample to cause a photodissociation of CO from the heme moiety, the intensity of the major band at 489 cm-1 was reduced as expected. On the other hand, the minor band at 523 cm-1 remained still obvious. It was suggested that the cytochrome b562-o complex may have an additional anomalous site for CO that is resistant to photodissociation. PMID- 2987232 TI - The effect of glucagon and adrenergic agonists on intracellular pH of isolated rat hepatocytes. AB - 19F NMR was used to measure intracellular [H+] of hepatocytes before and after incubation with glucagon and adrenergic agonists at their concentrations which give maximal stimulation of both glucose and urea production. Intracellular and extracellular pH was determined from the chemical shifts in resonances of alpha difluoromethylalanine. The alterations in intracellular [H+] with agonist treatment were, in all cases, found to be less than 0.1 pH unit in the pH range 6.7-7.8. It is concluded that changes in concentration of the intracellular [H+] do not play a significant role in the stimulation of urea and glucose production caused by these hormonal effectors. PMID- 2987233 TI - Iron metabolism in K562 erythroleukemic cells. AB - Iron delivery to K562 cells is enhanced by desferrioxamine through induction of transferrin receptors. Experiments were performed to further characterize this event with respect to iron metabolism and heme synthesis. In control cells, up to 85% of the iron taken up from iron-transferrin was incorporated into ferritin, 7% into heme, and the remainder into compartments not yet identified. In cells grown with desferrioxamine, net accumulation of intracellular desferrioxamine (14-fold) was observed and iron incorporation into ferritin and heme was inhibited by 86% and 75%, respectively. In contrast, complete inhibition of heme synthesis in cells grown with succinylacetone had no effect on transferrin binding or iron uptake. Exogenous hemin (30 microM) inhibited transferrin binding and iron uptake by 70% and heme synthesis by 90%. These effects were already evident after 2 h. Thus, although heme production could be reduced by desferrioxamine, succinylacetone, and hemin, cell iron uptake was enhanced only by the intracellular iron chelator. The effects of exogenous heme are probably unphysiologic and the greater inhibition of iron flow into heme can be explained by effects on early steps of heme synthesis. We conclude that in this cell model a chelatable intracellular iron pool rather than heme synthesis mediates regulation of iron uptake. PMID- 2987234 TI - Characterization of the cycle of iron-mediated electron transfer from Adriamycin to molecular oxygen. AB - The anticancer drug adriamycin binds iron and these complexes cycle to reduce molecular oxygen (Zweier, J. L. (1984) J. Biol. Chem. 259, 6056-6058). Optical absorption, EPR, and Mossbauer spectroscopic data are correlated with polarographic O2 consumption and chemical Fe2+ extraction measurements in order to characterize each step in this cycle. Fe3+ binds to adriamycin at physiologic pH forming a complex with an optical absorbance maximum at 600 nm. EPR signals at g = 4.2 and g = 2.01, and a doublet Mossbauer spectrum with isomer shift delta = 0.57 mm/s and quadrupole splitting delta EQ = 0.74 mm/s are observed indicating that the Fe3+ bound to adriamycin is high spin S = 5/2. Under anaerobic conditions the absorbance maximum at 600 nm decreases with an exponential decay constant = 0.77 h-1, and the EPR and Mossbauer spectra of Fe3+-adriamycin similarly decrease as the Fe3+ is reduced to EPR silent Fe2+. The Fe2+-adriamycin complex which is formed exhibits a Mossbauer spectrum with delta = 1.18 mm/s and delta EQ = 1.82 mm/s indicative of high spin Fe2+. As the EPR spectra of Fe3+ adriamycin decrease on reduction of the Fe3+ to Fe2+ a signal of the oxidized adriamycin free radical appears at g = 2.004 with line width of 8 G. On exposure to O2 the absorption maximum at 600 nm, the Fe3+ EPR, and the Fe3+ Mossbauer spectra all return. Polarographic measurements demonstrate that O2 is consumed and that H2O2 is formed. Addition of high affinity Fe2+ chelators block O2 consumption indicating that Fe2+ formation is essential for O2 reduction. This cycle of iron-mediated O2 reduction can explain the formation of the reactive reduced oxygen and adriamycin radicals which are thought to mediate the biological activity of adriamycin. PMID- 2987235 TI - Cross-resistance and biochemical studies with two classes of HeLa cell mutants resistant to cardiac glycosides. The unusual behavior of cardenolide SC4453. AB - In HeLa cells two different types of mutants resistant to the cardiac glycoside ouabain (OuaR mutants) or erythrophleum alkaloid cassaine (CasR mutants) have been obtained. One type of mutants resistant to these compounds (designated as group A) are highly resistant (between 50 and 2000-fold) to various cardiac glycosides and their genins such as ouabain, oleandrin, digitoxin, digitoxigenin, strophanthidin, convallatoxin, gitoxin, gitoxigenin, gitaloxin, bufalin, and digoxigenin, but exhibit no cross-resistance to SC4453, a digoxin analog which contains a pyridazine ring in place of the lactone ring in the C-17 position. The second type of mutants (group B) exhibit cross-resistance to all of the cardiac glycosides including SC4453, but their level of resistance is at least 5-10-fold less than that of group A mutants. Interestingly, both groups of mutants exhibited similar degree of cross-resistance towards digoxin and actodigin (AY22241), indicating some differences in their behavior from other cardiac glycosides. Both classes of mutants exhibit no cross-resistance to a wide variety of other structurally and functionally related compounds, e.g. sanguinarine nitrate, ethacrynic acid, penicillic acid, veratridine, harmaline hydrochloride, 5,5'-diphenylhydantoin, quindonium bromide, methyl quinolizinum bromide, estradiol 17 beta-acetate, 21-acetoxy-pregnenolone, vanadium pentoxide, digitonin, and adriamycin, indicating that the genetic lesions in both groups of mutants are specific for cardiac glycosides. This inference is supported by the observation that both group A and B mutants show reduced binding of [3H]ouabain. In group A mutants, a part of the Na+/K+-ATPase activity is highly resistant to inhibition by ouabain, indicating that the genetic lesion in these mutants directly affects Na+/K+-ATPase. In contrast, the Na+/K+-ATPase from the group B mutants showed similar resistance towards ouabain and SC4453 as observed for the parental HeLa cells, indicating that these mutants are affected in a cellular component, other than Na+/K+-ATPase, which is involved in the interaction of cardiac glycosides with the cells. The lack of cross-resistance of the group A mutants to SC4453 and normal sensitivity of their Na+/K+-ATPase to this compound provides strong evidence that the mechanism of interaction of SC4453 with Na+/K+ ATPase differs from that of other cardiac glycosides. PMID- 2987236 TI - Mossbauer studies of aconitase. Substrate and inhibitor binding, reaction intermediates, and hyperfine interactions of reduced 3Fe and 4Fe clusters. AB - Active beef heart aconitase contains a [4Fe-4S] cluster. One iron of the cluster, Fea, is labile and can be removed easily by oxidation in air to yield the [3Fe 4S]1+ cluster of inactive aconitase. We have previously shown that substrate binds to Fea. We have continued our Mossbauer studies by further investigating the active and inactive forms of the enzyme. When active aconitase, [4Fe-4S]2+, is mixed with substrate, two species (substrates or intermediates bound to Fea) labeled S1 and S2 are obtained. With the nitroanalogs of citrate and isocitrate, thought to be transition state analogs, and fluorocitrate, species S2, but not S1, is observed, suggesting that S2 represents a carbanion transition state complex. We have prepared Mossbauer samples by rapid mix/rapid freeze techniques. Using either citrate, isocitrate or cis-aconitate, the natural substrates, we have been able to detect at 0 degree C reaction intermediates in the 5-35 ms time range and, studying enzyme substrate interactions at subzero temperatures in a water/methanol/ethylene glycol solvent, we have observed new species when substrates were added at -60 degrees C. Details of these experiments are given, although in neither case can unique interpretations be offered at this time. We have also investigated reduced active aconitase ([4Fe-4S]1+; EPR at g = 1.94) in the presence of substrate with material selectively enriched with 57Fe in either Fea or the other three cluster sites. The spectra were analyzed with a spin Hamiltonian, and the results are discussed and interpreted in terms of three inequivalent Fe sites in the cluster. Finally, we have studied enzyme containing the reduced [3Fe-4S]0 cluster. There is no indication that citrate binds to the 3Fe cluster, and since no significant activity was observed, we conclude that aconitase containing a 3Fe cluster is not active in either oxidation state. PMID- 2987237 TI - Selective affinity chromatography with calmodulin fragments coupled to sepharose. AB - Calmodulin tryptic fragments 78-148, 107-148, and 1-77 coupled to Sepharose 4B were used to test the ability of different calmodulin-regulated enzymes to recognize different domains of calmodulin. Fragment 107-148, which contains a single Ca2+-binding domain, does not interact with any of the calmodulin binding proteins. Fragments 1-77 and 78-148, each of which contains two Ca2+-binding domains, have preserved their ability to interact with several calmodulin dependent enzymes. Most of the calmodulin-regulated enzymes in brain extracts, such as cAMP phosphodiesterase, cAMP-dependent protein kinase, and the calmodulin stimulated protein phosphatase (calcineurin) interact with fragment 78-148 in a Ca2+-dependent fashion. An ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N' tetraacetic acid-sensitive, calmodulin-independent, p-nitrophenyl phosphatase does not bind to the affinity column and is resolved from calcineurin at this step. Although calmodulin-stimulated protein kinase(s) can interact with fragment 78-148, their interaction is prevented by increased ionic strength even in the presence of Ca2+. Fragment 1-77 exhibits a higher degree of selectivity than fragment 78-148. Only cAMP-dependent protein kinase and cAMP phosphodiesterase bind to fragment 1-77. These results confirm the multiple modes of interaction of calmodulin with its target proteins and provide the basis for a selective purification of calmodulin-regulated enzymes by affinity chromatography on specific calmodulin fragments coupled to Sepharose. PMID- 2987238 TI - The human phosphoglycerate kinase multigene family. HLA-associated sequences and an X-linked locus containing a processed pseudogene and its functional counterpart. AB - Several phosphoglycerate kinase genes were previously detected in the human genome by blot hybridization with a phosphoglycerate kinase cDNA probe. Using subcloned fragments of the cDNA we estimate the presence of four independent phosphoglycerate kinase genes. These genes have been mapped to both the human X chromosome (band q13) and chromosome 6 (p12-21.1) using a panel of human-rodent somatic cell hybrids and by chromosomal in situ hybridization. The genomic distribution of phosphoglycerate kinase sequences is conserved in man and mouse, not only for the X chromosome, but also for linkage to the respective major histocompatibility complexes. Molecular cloning of X-linked phosphoglycerate kinase sequences led to the identification of a novel intronless phosphoglycerate kinase pseudogene which is localized proximal to the active gene on the X chromosome. PMID- 2987239 TI - In vitro maturation of circular bacteriophage P2 DNA. Purification of ter components and characterization of the reaction. AB - The protein components required for generation of cohesive ends in vitro from circular bacteriophage P2 DNA have been purified to near homogeneity. In the presence of ATP, the purified products of P2 genes M and P together with empty phage capsids (comprised primarily of the N protein) mediate site-specific cleavage of circular P2 DNA at the cohesive end site (cos). This terminase or ter system also utilizes circular DNAs of bacteriophages P4 and 186, introducing site specific scissions at cos sites within these molecules. The ter reaction exhibits a peculiar requirement for a circular DNA substrate. Substrate activity is greatly reduced when circular P2, P4, or 186 DNAs are linearized by restriction endonuclease hydrolysis. Furthermore, multimeric P4 DNA molecule sites are also essentially inactive in the linear form but are active in the circular state. The dependence of ter action on a circular substrate is not due to inhibition of the system by linear DNA, nor does it appear to reflect a requirement for substrate superhelicity since circular P4 DNA containing single strand scissions is subject to terminase action. The terminase reaction is supported by ATP, dATP, or beta, gamma-imido ATP, but not by other ribonucleoside triphosphates ADP, alpha, beta methylene ATP, or beta, gamma-methylene ATP. A DNA-dependent ATPase, which hydrolyzes ATP to AMP, copurifies with the P2 P protein and is inactivated with the same kinetics as P activity upon treatment with N-ethylmaleimide. The ATPase does not display specificity for P2 DNA in vitro. PMID- 2987240 TI - Subunit structure of a high-affinity receptor for type beta-transforming growth factor. Evidence for a disulfide-linked glycosylated receptor complex. AB - The structure of high-affinity receptors for type beta-transforming growth factor (beta TGF) has been examined by affinity labeling with 125I-beta TGF and disuccinimidyl suberate. The major receptor component labeled by 125I-beta TGF in mouse, rat, and chick fibroblasts migrated as a 280-290-kilodalton species on dodecyl sulfate-polyacrylamide electrophoresis gels in the presence of reductant, dithiothreitol. A larger (330-kilodalton) species was labeled in human fibroblasts, but comparative peptide mapping indicated a close structural relationship with receptors from mouse fibroblasts. In the absence of reductant, the affinity-labeled beta TGF receptor migrated in the gels as a larger disulfide linked complex. The molecular mass calculated from the hydrodynamic properties of native nonreduced beta TGF receptors was 565 (mouse) or 615 kilodaltons (human). Other molecular parameters for the beta TGF receptor were: Stokes radius, 8.3-8.5 nm; sedimentation coefficient, 12.7-13.0 S; and frictional ratio, f/f0 = 1.4. The beta TGF receptor was solubilized under conditions in which the structural and ligand-binding properties of the native state were retained. beta TGF receptors solubilized from human, mouse, and chick cells interacted specifically with immobilized wheat germ agglutinin. These data suggest that the high affinity receptor for beta TGF in human, rodent, and avian fibroblasts is a disulfide linked glycosylated 565-615-kilodalton complex with a 280-330-kilodalton subunit that contains the ligand-binding site. The oligomeric structure of the beta TGF receptor does not appear to be induced by receptor occupancy with the ligand. PMID- 2987241 TI - The gene for the beta subunit of bovine luteinizing hormone encodes a gonadotropin mRNA with an unusually short 5'-untranslated region. AB - Both cDNA and genomic clones encoding the beta subunit of bovine luteinizing hormone (LH) have been isolated and characterized. The nucleotide sequence was determined for the entire gene and 776 base pairs of 5'-flanking sequence. The mRNA cap site and polyadenylation site were mapped by primer extension and S1 nuclease protection, respectively. The bovine LH beta spans less than 1.1 kilobase pairs and has three exons encoding a 550 nucleotide mRNA (excluding the poly(A) tail). Bovine LH beta is a single-copy gene, in contrast to human LH beta, which is a member of the LH/chorionic gonadotropin beta subunit multigene family. Comparison of the bovine LH beta gene with the human LH beta/chorionic gonadotropin gene family reveals a high degree of nucleotide sequence homology, both within the genes and in the 5'-flanking sequences. Despite this extensive sequence conservation, there is a major difference between the two species in the selection of a promoter site. As a result, the bovine LH beta gene produces an mRNA with an usually short 5'-untranslated region of only 6-11 nucleotides. PMID- 2987242 TI - Thrombin induces serotonin secretion and aggregation independently of inositol phospholipids hydrolysis and protein phosphorylation in human platelets permeabilized with saponin. AB - We have observed that the addition of Ca2+ to platelets, permeabilized with saponin, promotes a drastic dephosphorylation of proteins and polyphosphoinositides without inducing platelet responses. Subsequent addition of thrombin could promote secretion of serotonin and aggregation in the absence of phospholipase C-induced breakdown of the inositol phospholipids and protein phosphorylation. This information indicates that activation of saponized platelets by thrombin is independent of the formation of second messengers derived from the phospholipase C-induced breakdown of the inositol phospholipids. The implications of this result for intact platelets are discussed. PMID- 2987243 TI - Phosphorylation of the mammalian beta-adrenergic receptor by cyclic AMP-dependent protein kinase. Regulation of the rate of receptor phosphorylation and dephosphorylation by agonist occupancy and effects on coupling of the receptor to the stimulatory guanine nucleotide regulatory protein. AB - In some systems, such as the turkey erythrocyte, agonist-promoted phosphorylation of the beta-adrenergic receptor appears to be associated with desensitization of the adenylate cyclase system. This process can be partially mimicked by cyclic AMP analogs. Accordingly, we have investigated the phosphorylation of the pure mammalian beta-adrenergic receptor by the pure catalytic subunit of the cyclic AMP-dependent protein kinase. The beta-adrenergic receptor, purified from hamster lung to apparent homogeneity, contains a single polypeptide of Mr approximately 64,000. The receptor can be phosphorylated in vitro by the catalytic subunit of cyclic AMP-dependent protein kinase (approximately 2 mol of phosphate (on serine residues) per mol). Isoproterenol, a beta-agonist, promoted a 2-3-fold increase in the rate of receptor phosphorylation which was blocked by the beta-antagonists propranolol and alprenolol. High performance liquid chromatographic tryptic peptide mapping reveals two major phosphorylation sites. Phosphorylated receptor can be completely dephosphorylated by a high molecular weight phosphoprotein phosphatase. The rate of receptor dephosphorylation is enhanced 2-3-fold by isoproterenol and this effect is blocked by alprenolol. The functional significance of receptor phosphorylation was examined using ligand binding and reconstitution techniques. While the binding of isoproterenol and alprenolol to the receptor was unaffected by phosphorylation, the ability of the receptor to interact with the stimulatory guanine nucleotide regulatory protein, as assessed by isoproterenol-promoted GTPase activity, was decreased 24 +/- 1% (mean +/- S.E., p less than 0.001, n = 17). The quantitative extent of receptor phosphorylation and functional impairment are virtually identical to those previously observed when intact turkey erythrocytes were incubated with cyclic AMP. These data provide a direct demonstration of regulation of the function of the isolated beta-adrenergic receptor by cyclic AMP-dependent protein kinase. PMID- 2987244 TI - Transcriptional organization of the convergent overlapping dnaQ-rnh genes of Escherichia coli. AB - The transcriptional organization was determined for the DNA polymerase III epsilon subunit (dnaQ) and the ribonuclease H (rnh) genes, which are closely spaced and organized in a face-to-face system (Maki, H., Horiuchi, T., and Sekiguchi, M. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 7137-7141). Nuclease S1 mapping revealed that transcription starts from a single promoter for rnh and two promoters for dnaQ and proceeds in opposite directions. The 5' terminus of the rnh RNA overlaps about 100 and 20 nucleotide sequences with the 5' terminus of the dnaQ-1 and dnaQ-2 RNA, respectively. The levels of in vivo transcription for the three RNA species increased altogether by more than 10-fold in cells carrying a multicopy plasmid with intact dnaQ-rnh genes, keeping the relative level for the convergent transcription units nearly constant. PMID- 2987245 TI - Kinetics of reduction of cytochrome c oxidase by dithionite and the effect of hydrogen peroxide. AB - The reduction of cytochrome c oxidase by dithionite was reinvestigated with a flow-flash technique and with varied enzyme preparations. Since cytochrome a3 may be defined as the heme in oxidase which can form a photolabile CO adduct in the reduced state, it is possible to follow the time course of cytochrome a3 reduction by monitoring the onset of photosensitivity. The onset of photosensitivity and the overall rate of heme reduction were compared for Yonetani and Hartzell-Beinert preparations of cytochrome c oxidase and for the enzyme isolated from blue marlin and hammerhead shark. For all of these preparations the faster phase of heme reduction, which is dithionite concentration-dependent, is almost completed when the fraction of photosensitive material is still small. We conclude that cytochrome a3 in the resting enzyme is consistently reduced by an intramolecular electron transfer mechanism. To determine if this is true also for the pulsed enzyme, we examined the time course of dithionite reduction of the peroxide complex of the pulsed enzyme. It has been previously shown that pulsed cytochrome c oxidase can interact with H2O2 and form a stable room temperature peroxide adduct (Bickar, D., Bonaventura, J., and Bonaventura, C. (1982) Biochemistry 21, 2661-2666). Rather complex kinetics of heme reduction are observed when dithionite is added to enzyme preparations that contain H2O2. The time courses observed provide unequivocal evidence that H2O2 can, under these conditions, be used by cytochrome c oxidase as an electron acceptor. Experiments carried out in the presence of CO show that a direct dithionite reduction of cytochrome a3 in the peroxide complex of the pulsed enzyme does not occur. PMID- 2987246 TI - A strong sequence homology exists between the major RNA polymerase sigma factors of Bacillus subtilis and Escherichia coli. AB - The Bacillus subtilis rpoD gene has been sequenced and the primary structure of its product deduced. The molecular weight calculated for the sigma 43 is 42,828. The DNA and protein sequences of sigma 43 exhibit extensive homology to the Escherichia coli rpoD gene and its sigma 70 product, especially in the C-terminal two-thirds of the sequence. Other proteins exhibiting partial homology with sigma 43 include the E. coli nusA protein, the E. coli htpR (heat-shock regulatory gene) product sigma 32, and specific DNA-binding proteins. No amino acid homology was found between sigma 43 and B. subtilis phage SP01 sigma gp28, phage T7 RNA polymerase, or E. coli DNA primase. The gene exhibits a strong ribosomal binding site and a typical rho-independent transcription terminator sequence. A typical transcription terminator sequence was not observed upstream from the sigma 43 gene. The sigma 43 gene may be part of an operon, resembling the situation found in the E. coli sigma operon. PMID- 2987247 TI - Purification and characterization of a paired basic residue-specific pro opiomelanocortin converting enzyme from bovine pituitary intermediate lobe secretory vesicles. AB - Pro-opiomelanocortin (adrenocorticotropin/endorphin prohormone) is processed to yield active hormones by cleavages at paired basic amino acid residues. In this study, an enzyme that specifically cleaves at the paired basic residues of this prohormone has been purified from bovine pituitary intermediate lobe secretory vesicles, the intracellular processing site of proopiomelanocortin. This enzyme, named pro-opiomelanocortin converting enzyme, has been characterized as a glycoprotein of Mr approximately 70,000. It has an apparent isoelectric point between 3.5 and 4.0. The pH optimum of the pro-opiomelanocortin converting enzyme is between 4 and 5, but the enzyme is highly active at the intravesicular pH of 5.1-5.6. The enzyme specifically cleaved the Lys-Arg pairs of pro opiomelanocortin to yield Mr = to 21,000-23,000 ACTH, beta-lipotropin, Mr 13,000 and 4,500 ACTH, beta-endorphin, and a Mr = 16,000 NH2-terminal glycopeptide, the products synthesized by the pituitary intermediate lobe in situ. NH2- and COOH terminal analysis of the products indicated that the pro-opiomelanocortin converting enzyme cleaves the peptide bond either between the Lys and Arg or on the carboxyl side of the Arg at Lys-Arg pairs of pro-opiomelanocortin. The intracellular localization, pH optimum, and cleavage specificity of the enzyme suggest that it may function as a pro-opiomelanocortin processing enzyme in the pituitary intermediate lobe in vivo. PMID- 2987248 TI - Characterization of the lep operon of Escherichia coli. Identification of the promoter and the gene upstream of the signal peptidase I gene. AB - The DNA sequence flanking the gene (lep) encoding signal peptidase I of Escherichia coli has been determined. The upstream flanking sequence contains a gene (lepA) that encodes a polypeptide of 598 amino acid residues and terminates 18 base pairs from the initiation codon of the lep gene. The position of the lep promoter was determined by both gene fusion with the lacZ gene as well as by S1 nuclease mapping of the lep mRNA to be 73 base pairs upstream from the initiation codon of the lepA gene. The lepA gene was cloned into a high expression vector (pIN-III), and its gene product was identified to be a protein of apparent molecular weight of 76,000. This gene product was preferentially localized in the cytoplasmic membrane and periplasmic fractions upon subcellular fractionation. The DNA sequence immediately downstream of the lep gene contains features consistent with a rho factor-independent transcriptional termination site, indicating that the lep operon encodes only two proteins (lepA and lep). PMID- 2987249 TI - Biosynthetic precursors and in vitro translation products of the glucose transporter of human hepatocarcinoma cells, human fibroblasts, and murine preadipocytes. AB - Antisera to the human erythrocyte Glc transporter immunoblotted a polypeptide of Mr 55,000 in membranes from human hepatocarcinoma cells, Hep G2, human fibroblasts, W138, and murine preadipocytes, 3T3-L1. This antisera immunoprecipitated the erythrocyte protein which had been photoaffinity labeled with [3H]cytochalasin B, immunoblotted its tryptic fragment of Mr 19,000, and immunoblotted the deglycosylated protein as a doublet of Mr 46,000 and 38,000. This doublet reduced to a single polypeptide of Mr 38,000 after boiling. When Hep G2, W138, and 3T3-L1 cells were metabolically labeled with L-[35S]methionine for 6 h, a broad band of Mr 55,000 was immunoprecipitated from membrane extracts. In pulse-chase experiments, two bands of Mr 49,000 and 42,000 were identified as putative precursors of the mature transporter. The t1/2 for mature Glc transporter was 90 min for Hep G2 cells that had been starved for methionine (2 h) and pulsed for 15 min with L-[35S]methionine. Polypeptides of Mr 46,000 and 38,000 were immunoprecipitated from Hep G2 cells that had been metabolically labeled with L-[35S]methionine in the presence of tunicamycin. This doublet reduced to the single polypeptide of Mr 38,000 after boiling. In the absence of tunicamycin, but not in its presence, mature polypeptide of Mr 55,000 was immunoprecipitated from Hep G2 cells metabolically labeled with D-[3H]GlcN. A polypeptide of Mr 38,000 was observed in boiled immune complexes from the in vitro translation products of Hep G2, W138, and 3T3-L1 cell RNA. Dog pancreatic microsomes cotranslationally, but not posttranslationally, converted this to a polypeptide of Mr 35,000. A model for Glc transporter biogenesis is proposed in which the primary translation product of Mr 38,000 is converted by glycosylations to a polypeptide of Mr 42,000. The latter is then processed via heterogeneous complex N-linked glycosylations to form the mature Glc transporter, Mr 55,000. PMID- 2987250 TI - 23Na NMR studies of rat outer medullary kidney tubules. AB - Two reservations have previously made interpretation of biological 23Na NMR measurements difficult: the "size" of the extracellular space penetrated by the shift reagent and the possibility of a 60% reduction in the intensity of the NMR visible 23Na signal due to quadrupolar interactions (Berendsen, H. J. C., and Edzes, H. T. (1973) Ann. N. Y. Acad. Sci. 204, 459-485; Civan, M. M., Degani, H., Margalit, Y., and Shporer, M. (1983) Am. J. Physiol. 245, C213-C219; Gupta, R. K., and Gupta, P. (1982) J. Magn. Reson. 47, 344-350). We have addressed both these issues using a suspension of rat outer medullary kidney tubules, nephron segments responsible for the fine control of total body volume and electrolyte balance. First, the extracellular space penetrated by the shift reagent dysprosium tripolyphosphate, as defined by the extracellular 23Na resonance, revealed a space similar to that which contained extracellular 35Cl- ions. Measurement of an extracellular 35Cl- space using 35Cl NMR was possible because the intracellular 35Cl- resonance was broadened beyond detection in the cells studied. Second, to characterize the reduction of the 23Na signal by quadrupolar interactions, the intracellular 23Na level was raised artificially by simultaneously inhibiting Na+ efflux and increasing the ion permeability of the plasma membrane. Under these conditions, NMR-observable intracellular Na+ reached a level which was approximately 81% of that in the medium, a level determined using chemical techniques. This observation would suggest that the resonance of the intracellular 23Na pool was not subject to a 60% reduction in signal intensity, as a result of nuclear quadrupolar interaction. The intracellular 23Na level measured, under basal conditions, was 23 +/- 2 mumol/ml of cell water (37 degrees C) (n = 3, S.D.) and was demonstrated to be responsive to a number of physiological stimuli. The level was temperature-sensitive. It was reduced by inhibitors of apical Na+ transport, furosemide and amiloride, and it was raised with (Na+ + K+)-ATPase inhibition. The furosemide and amiloride actions described would suggest that the Na+-transporting mechanisms sensitive to these agents (e.g. Na+/K+/Cl- cotransport system, Na+:H+ exchange system) contribute to the regulation of the intracellular Na+ level in the kidney tubular preparation studied. PMID- 2987251 TI - Purification and structure of the intact Ada regulatory protein of Escherichia coli K12, O6-methylguanine-DNA methyltransferase. AB - The ada gene of Escherichia coli K12, the regulatory gene for the adaptive response of bacteria to alkylating agents, was cloned and placed under the control of the lac regulatory region on a multicopy runaway plasmid, thereby yielding a hybrid plasmid pYN3059. Ada protein with a molecular weight of about 38,000 was overproduced when cells harboring pYN3059 were incubated at 42 degrees C in the presence of a lac inducer, isopropyl-beta-D-thiogalactoside. Taking advantage of overproduction of Ada protein, we purified the protein to apparent physical homogeneity. The purified 38,000-dalton Ada protein transferred the methyl group from the O6-methylguanine residue of alkylated DNA to the Ada protein, per se. Although the Ada protein was degraded to smaller polypeptides when crude extracts or partially purified preparations were incubated in a high ionic-strength buffer at neutral pH, the purified Ada protein remained stable under the same conditions, indicating that the Ada protein may not undergo autodegradation. An amino-terminal sequence and total amino acid composition of the purified Ada protein were in accord with nucleotide sequence of the ada gene, determined by the dideoxy method using M13 phage. It was deduced that Ada protein comprises 354 amino acids and its molecular weight is 39,385. The promoter for the ada gene was determined by S1 nuclease mapping. PMID- 2987252 TI - Stimulation of rat ovarian cell steroidogenesis by high density lipoproteins modified with tetranitromethane. AB - Human high density lipoprotein (devoid of apo-E) was modified by nitration of tyrosine residues with tetranitromethane. As a result of extensive cross-linking, monomeric apo-A-I was markedly depleted, as assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the modified HDL did not effectively bind to high-affinity sites present on dispersed rat ovarian cells and isolated rat ovarian membranes. Nonetheless, the modified HDL retained the ability to stimulate steroidogenesis by both dispersed rat ovarian cells and cultured rat granulosa cells to a degree at least equal to that of native HDL. Modified HDL stimulated luteal steroidogenesis under basal conditions and when cells were stimulated with luteinizing hormone or 8-bromo-cAMP. Although modified HDL did not effectively bind to high-affinity sites, it exhibited substantial "nonspecific" or "low-affinity" binding which was not displaceable by native HDL. These data suggest that high-affinity binding is not an essential event in the "HDL pathway" and that HDL can deliver its sterols through low-affinity cellular associations. PMID- 2987253 TI - Mechanisms of detergent effects on membrane-bound (Na+ + K+)-ATPase. AB - Because the nonionic detergent octaethylene glycol dodecyl ether has been used extensively for studies on active solubilized preparations of (Na+ + K+)-ATPase, we tried to see if the detergent alters the properties of the membrane-bound enzyme prior to solubilization. Addition of the detergent, at concentrations below its critical micellar concentration, to reaction mixtures containing the highly purified membrane-bound enzyme reduced the K0.5 of ATP for (Na+ + K+) dependent ATPase activity without affecting the maximal velocity or abolishing the negative cooperativity of the substrate-velocity curve. Under these conditions, however, the enzyme was not solubilized as evidenced by complete sedimentation of the membrane fragments containing the enzyme upon centrifugation at 100,000 X g for 30 min. Other nonsolubilizing effects of the detergent included an increase in K0.5 of K+, inhibition of Na+-dependent ATPase with no effect on K0.5 of ATP for this activity, and reductions in the spontaneous decomposition rates of the K+-sensitive phosphoenzyme obtained from ATP and the phosphoenzyme obtained from Pi. The nonsolubilizing effects of the detergent on the purified enzyme were obtained with no detectable lag, were readily reversible, and could be distinguished from its vesicle-opening effects on crude membrane preparations. Several other nonionic and ionic detergents had similar effects on the enzyme. The findings indicate (a) detergent binding to hydrophobic sites on extramembranous segments of enzyme subunits; (b) that occupation of these sites mimics the effects of ATP at a low-affinity regulatory site with no effect on high-affinity ATP binding to the catalytic site; and (c) that in studies on detergent-solubilized preparations, it is necessary to distinguish between the effects of solubilization per se and detergent effects at the regulatory site. PMID- 2987255 TI - Effects of macrophage profilin on actin in the presence and absence of acumentin and gelsolin. AB - We purified profilin from rabbit alveolar macrophages and documented its structural and functional similarity to profilins isolated from other cells. The KD for formation of the macrophage profilin-actin complex was 3.0 +/- 0.8 microM (mean +/- S.D.). The affinity of this protein for actin did not change significantly in the presence of various concentrations of KCl and MgCl2, profilin-actin complex concentration being strictly dependent on the critical actin monomer concentration and free profilin concentration. We also examined profilin's interactions with actin in the presence of acumentin, a macrophage protein which inhibits actin monomer exchange at the "pointed" ends of actin filaments. Low concentrations of this protein caused substantial decreases in estimated profilin-actin complex concentration. The macrophage gelsolincalcium ion complex which blocks exchange at the "barbed" end of actin filaments, when added to profilin and actin solutions in substoichiometric concentrations, caused large increases in estimated profilin-actin complex concentration. The changes in calculated profilin-actin complex concentration induced by these two actin modulating proteins were too large to be explained solely by their effects on critical actin monomer concentration. PMID- 2987254 TI - Magnetic circular dichroism studies of succinate dehydrogenase. Evidence for [2Fe 2S], [3Fe-xS], and [4Fe-4S] centers in reconstitutively active enzyme. AB - Reconstitutively active and inactive succinate dehydrogenase have been investigated by low temperature magnetic circular dichroism (MCD) and EPR spectroscopy and room temperature CD and absorption spectroscopy. Reconstitutively active succinate dehydrogenase is found to contain three spectroscopically distinct Fe-S clusters: S1, S2, and S3. In agreement with previous studies, MCD and CD spectroscopy confirm that center S1 is a succinate reducible [2Fe-2S]2+,1+ center. The MCD characteristics of center S2 identify it as a dithionite-reducible [4Fe-4S]2+,1+ similar to those in bacterial ferredoxins. EPR power saturation studies and the weakness of the EPR signal from reduced S2 indicate that there is a weak magnetic interaction between centers S1 and S2 in their paramagnetic, S = 1/2, reduced states. Center S3 is identified both by the form of the MCD spectrum and the characteristic magnetization behavior as a reduced [3Fe-xS] center in both succinate- and dithionite-reduced reconstitutively active succinate dehydrogenase. Arguments are presented in favor of centers S2 and S3 being separate centers rather than interconversion products of the same cluster. Reconstitutively inactive succinate dehydrogenase is found to be deficient in center S3. These results resolve many of the controversies concerning the Fe-S cluster content of succinate dehydrogenase and reconcile published EPR data with analytical and core extrusion studies. Moreover, they indicate that center S3 is a necessary requirement for reconstitutive activity and suggest that it is able to sustain ubiquinone reductase activity as a [3Fe xS] center. PMID- 2987256 TI - Rat intestinal nucleotide-sugar pyrophosphatase. Localization, partial purification, and substrate specificity. AB - The nucleotide-sugar pyrophosphatase activity of rat small intestine was studied using GDP-[14C]Man as substrate. The highest specific activities in the gastrointestinal tract were in the proximal small intestine, with a preferential localization in villus tip cells. Purified brush-border membranes were highly enriched in nucleotide-sugar pyrophosphatase. After the enzyme was solubilized with detergent and purified 180-fold, it hydrolyzed FAD and p-nitrophenyl-5' thymidylate, as well as nucleotide sugars. That the same enzyme, a 5'-nucleotide phosphodiesterase, is responsible for nucleotide-sugar pyrophosphatase, phosphodiesterase I, and FAD pyrophosphatase activities is indicated by: co migration in electrophoresis, parallel thermal inactivation, competitive inhibition studies, and similar regional, cellular, and subcellular localizations. PMID- 2987258 TI - The sugar phosphate specificity of rat hepatic 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase. AB - The sugar phosphate specificity of the active site of 6-phosphofructo-2-kinase and of the inhibitory site of fructose-2,6-bisphosphatase was investigated. The Michaelis constants and relative Vmax values of the sugar phosphates for the 6 phosphofructo-2-kinase were: D-fructose 6-phosphate, Km = 0.035 mM, Vmax = 1; L sorbose 6-phosphate, Km = 0.175 mM, Vmax = 1.1; D-tagatose 6-phosphate, Km = 15 mM, Vmax = 0.15; and D-psicose 6-phosphate, Km = 7.4 mM, Vmax = 0.42. The enzyme did not catalyze the phosphorylation of 1-O-methyl-D-fructose 6-phosphate, alpha- and beta-methyl-D-fructofuranoside 6-phosphate, 2,5-anhydro-D-mannitol 6 phosphate, D-ribose 5-phosphate, or D-arabinose 5-phosphate. These results indicate that the hydroxyl group at C-3 of the tetrahydrofuran ring must be cis to the beta-anomeric hydroxyl group and that the hydroxyl group at C-4 must be trans. The presence of a hydroxymethyl group at C-2 is required; however, the orientation of the phosphonoxymethyl group at C-5 has little effect on activity. Of all the sugar monophosphates tested, only 2,5-anhydro-D-mannitol 6-phosphate was an effective inhibitor of the kinase with a Ki = 95 microM. The sugar phosphate specificity for the inhibition of the fructose-2,6-bisphosphatase was similar to the substrate specificity for the kinase. The apparent I0.5 values for inhibition were: D-fructose 6-phosphate, 0.01 mM; L-sorbose 6-phosphate, 0.05 mM; D-psicose 6-phosphate, 1 mM; D-tagatose 6-phosphate, greater than 2 mM; 2,5 anhydro-D-mannitol 6-phosphate, 0.5 mM. 1-O-Methyl-D-fructose 6-phosphate, alpha- and beta-methyl-D-fructofuranoside 6-phosphate, and D-arabinose 5-phosphate did not inhibit. Treatment of the enzyme with iodoacetamide decreased sugar phosphate affinity in the kinase reaction but had no effect on the sensitivity of fructose 2,6-bisphosphatase to sugar phosphate inhibition. The results suggest a high degree of homology between two separate sugar phosphate binding sites for the bifunctional enzyme. PMID- 2987257 TI - Parietal cell protein kinases. Selective activation of type I cAMP-dependent protein kinase by histamine. AB - cAMP-dependent protein kinases have been characterized in parietal cells isolated from rabbit gastric mucosa. Both Type I and Type II cAMP-dependent protein kinase isozymes are present in these cells. Type II isozymes were detected in 900, 14,000, and 100,000 X g particulate fractions as well as 100,000 X g cytosolic fractions; Type I isozymes were found predominately in the cytosolic fraction. When parietal cells were stimulated with histamine, an agent that elevates intracellular cAMP content and initiates parietal cell HCl secretion, cAMP dependent protein kinase activity was increased in homogenates of these cells as measured by an increase in the cAMP-dependent protein kinase activity ratio. Histamine activation of cAMP-dependent protein kinase was correlated with parietal cell acid secretory responses which were measured indirectly as increased cellular uptake of the weak base, [14C]aminopyrine. These results suggest that cAMP-dependent protein kinase(s) is involved in the control of parietal cell HCl secretion. The parietal cell response to histamine may be compartmentalized because histamine appears to activate only a cytosolic Type I cAMP-dependent protein kinase isozyme, as determined by three different techniques including 1) ion exchange chromatography; 2) Sephadex G-25 to remove cAMP and allow rapid reassociation of the Type II but not the Type I isozyme; and 3) 8-azido-[32P]cAMP photoaffinity labeling. Forskolin, an agent that directly stimulates adenylate cyclases, was found to activate both the Type I and Type II isozymes. Several cAMP-dependent protein kinases were also detected in parietal cell homogenates, including a Ca2+-phospholipid-sensitive or C kinase and two casein kinases which were tentatively identified as casein kinase I and II. At least two additional protein kinases with a preference for serine or lysine-rich histones, respectively, were also detected. The function of these enzymes in parietal cells remains to be shown. PMID- 2987260 TI - Adenine nucleotide translocase activity and sensitivity to inhibitors in hepatomas. Comparison of the ADP/ATP carrier in mitochondria and in a purified reconstituted liposome system. AB - Adenine nucleotide uptake was found to be lower in mitochondria from hepatoma 7777, 7800, and 9618A than in the host livers. Moreover, in the fast-growing hepatoma 7777 the sensitivity of the adenine nucleotide translocase to inhibition by carboxyatractylate and bongkrekic acid was considerably decreased. Purification of the ADP/ATP carrier from hepatoma 7777 mitochondria and its reconstitution into an artificial liposome system reversed the abnormal kinetics in that the adenine nucleotide uptake and response to inhibitors were identical in proteoliposome preparations from host liver and tumor mitochondria. Analysis of the lipids of the hepatoma inner mitochondrial membrane indicated considerable differences from normal in the levels of phospholipids and cholesterol. Most striking was the increase in cholesterol and sphingomyelin of the hepatoma 7777 inner membrane. An artificial liposome system containing cholesterol in addition to the standard phospholipids could produce alterations in kinetics of the purified ADP/ATP carrier from heart mitochondria similar to those seen in the hepatoma 7777. In general, these results support the suggestion that alterations in the lipid environment of the inner mitochondrial membrane rather than intrinsic changes in the carrier protein itself produce the aberrant observations of adenine nucleotide translocase activity in hepatoma mitochondria. PMID- 2987259 TI - Structure-function relationships in glucagon. Re-evaluation of glucagon-(1-21). AB - Glucagon-(1-21) was prepared fully synthetically as well as by carboxypeptidase A digestion of natural porcine glucagon. Neither of the two preparations had glucagon agonistic effects with regard to receptor binding or adenylate cyclase activation in purified rat liver plasma membranes. Nor did these preparations contain lipolytic activity in isolated free fat cells. A preliminary batch of glucagon-(1-21) prepared by carboxypeptidase A digestion did, however, contain 1 2% glucagon bioactivity. This activity was separated from glucagon-(1-21) by high performance liquid chromatography and quantitatively recovered in four minor hind peaks which eluted close to but not in a position identical to the elution position of native glucagon. PMID- 2987261 TI - Regulation of internal pH of sea urchin sperm. A role for the Na/K pump. AB - In the absence of sodium, sea urchin sperm have an acidic internal pH. The addition of sodium, lithium, or ammonium, but not of potassium ions, induces an internal alkalization. If potassium is added in the presence of sodium, a further alkalization is obtained; in contrast, potassium addition in presence of Li+ or NH+4 does not change the internal pH. The K+-induced pHi change is inhibited by ouabain and when sperm are depleted of their ATP. A large part of the potassium influx is stimulated by Na+, but not Li+, and inhibited by ouabain and cellular ATP depletion. We conclude that activity of Na/K-ATPase pumps located in the plasma membrane of sea urchin sperm could play a role in regulating the internal pH of sea urchin sperm by recycling sodium ions that enter the cell through Na/H countermovements. PMID- 2987262 TI - Poliovirus replicase stimulation by terminal uridylyl transferase. AB - In an in vitro poliovirus replication system, purified viral polymerase, plus sense virion RNA, and a host factor have been previously shown to be necessary for the transcription of minus strands. We have found that a partially purified eukaryotic initiation factor-2 (eIF-2) fraction from rabbit reticulocytes can replace HeLa host factor in the replicase reaction. This enzyme preparation contains eIF-2 and two other major proteins. In addition to eIF-2 activity, which does not appear to play a role in the replicase reaction, we find that the fraction contains terminal uridylyl transferase activity. The enzyme adds UMP moieties to the 3' end of primer RNA molecules. The number of UMP residues added depends on the primer. Although long tails of heterogeneous lengths (50 to 100 nucleotides) can be polymerized on the 3' end of oligo(U), a poly(A) primer accepts only four U's. The terminal uridylyl transferase activity requires only UTP, Mg2+, a sulfhydryl reagent, and an RNA primer for activity. It is partially associated with ribosomes. We provide preliminary evidence that it may be responsible for host factor-like activity. We present a model for minus strand synthesis by poliovirus replicase, based on the hypothesis that a terminal uridylyl transferase can participate in initiation. PMID- 2987263 TI - Purified rat hepatic beta 2-adrenergic receptor. Structural similarities to the rat fat cell beta 1-adrenergic receptor. AB - The mammalian beta 2-adrenergic receptor from rat liver has been purified by sequential cycles of affinity chromatography followed by steric exclusion high performance liquid chromatography. In purified preparations, the overall yield of receptor approaches 10%. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of highly purified receptor preparations reveals a single peptide, Mr = 67,000, as judged by silver staining. Purified beta 2-adrenergic receptor migrates on steric-exclusion high performance liquid chromatography in two peaks, Mr = 140,000 and 67,000. Specific binding of (-)-[3H]dihydroalprenolol and (-)-[125I]iodocyanopindolol to purified rat liver beta-adrenergic receptor preparations is stereoselective and displays a rank order of potencies characteristic of a beta 2-adrenergic receptor. The mammalian beta 1-adrenergic receptor of rat fat cells has also been purified (Cubero, A., and Malbon, C.C. (1984) J. Biol. Chem. 259, 1344-1350). When purified in the presence of protease inhibitors, radioiodinated beta 1-adrenergic receptors from rat fat cells and beta 2-adrenergic receptors from rat liver comigrate on sodium dodecyl sulfate polyacrylamide gels as 67,000 Mr peptides. Autoradiograms of two-dimensional partial proteolytic digests of the purified, radioiodinated rat liver beta adrenergic receptor, as generated by alpha-chymotrypsin, Staphylococcus aureus V8 protease, and elastase reveal a pattern of peptide fragments essentially identical to those generated by partial proteolytic digests of the purified radioiodinated beta 1-receptor from rat fat cells. This data suggests that a high degree of homology exists between these two pharmacologically distinct mammalian beta-adrenergic receptor proteins. PMID- 2987265 TI - Preferential localization of variant nucleosomes near the 5'-end of the mouse dihydrofolate reductase gene. AB - We have probed the structure of nucleosomes within the 31-kilobase pair long, transcriptionally active gene for dihydrofolate reductase (DHFR) in mouse cells which contain multiple copies of the DHFR gene. We found that the distribution of electrophoretically variant nucleosomes within the DHFR gene is highly nonuniform: variant DHFR nucleosomes are abundant within and in the immediate vicinity of the approximately 200-base pair (bp) long first DHFR exon, and decrease by at least 10-fold within two nucleosomes upstream and downstream from this region. The nonuniformly distributed variant DHFR mononucleosomes are of two electrophoretically distinguishable discrete types. One corresponds to a mononucleosome containing a approximately 180-bp DNA fragment and possibly also histone H1 and high mobility group proteins. The other type of variant DHFR mononucleosome contains a approximately 146-bp DNA fragment, and its changes in relative content within the DHFR gene closely parallel those of the 180-bp variant mononucleosome. Several lines of evidence are consistent with the interpretation that the electrophoretically variant approximately 146-bp (core) mononucleosome species corresponds to diubiquitinated DHFR nucleosomes. We discuss possible causal relationships between the observed nonuniform distribution of variant nucleosomes within the DHFR gene and the DHFR gene transcription. PMID- 2987264 TI - Transcriptional regulation of mouse dihydrofolate reductase in the cell cycle. AB - We have studied transcription of the dihydrofolate reductase gene as cells progress through the cell cycle. We used the techniques of DNA-excess filter hybridization and in vivo pulse labeling of a synchronous population of cells to measure the transcription rate of the dihydrofolate reductase gene in different phases of the cell cycle. Our results indicate that the dihydrofolate reductase gene is cell cycle regulated at the transcriptional level. The transcription rate is low in G1, increases 7-fold at the beginning of S phase, decreases almost immediately thereafter, and remains low throughout the remainder of S and into G2. This cell cycle regulation seen in the G1 to S phase transition is achieved by increasing the rate of transcription from a single promoter region. The fact that this region is similar to promoter regions of other housekeeping genes suggests that this type of regulation may not be unique to dihydrofolate reductase. PMID- 2987266 TI - Analysis of adenosine kinase mutants of baby hamster kidney cells using affinity purified antibody. AB - The adenosine kinase enzymes from arabinofuranosyladenine-resistant (araAr) mutants of the baby hamster kidney cell line were analyzed using adenosine kinase specific antibody probes purified by adenosine kinase-Sepharose column chromatography. Wild-type baby hamster kidney cells were shown to produce two adenosine kinase polypeptides of Mr 43,000 and 40,000. The class I araAr mutants that have no detectable adenosine kinase activity are completely deficient in the two adenosine kinase polypeptides. As expected, the class II araAr mutants, which had been shown to have an altered ribonucleotide diphosphate reductase activity, produce a wild-type level of the two adenosine kinase polypeptides. The five class III araAr mutants which are adenosine-sensitive (AdoS) have various levels of adenosine kinase activity and produced two adenosine kinase polypeptides with similar Mr as that of wild-type cells. The adenosine kinase proteins synthesized by two of the AdoS mutants, ara-19a and ara-74b, differed from wild type in their isoelectric points. These results plus the observations that the AdoS mutants produce adenosine kinase enzymes with altered kinetic properties suggest a point mutation in the adenosine kinase gene. An araAr mutant, ara-60a, with intermediate adenosine sensitivity, was shown to have two truncated adenosine kinase polypeptides. This observation strongly supports the genetic data which suggests that there is only one functional adenosine kinase allele in baby hamster kidney cells and that the two adenosine kinase polypeptides are due to posttranscriptional modification. PMID- 2987267 TI - Characterization of a calmodulin-dependent protein phosphatase from human platelets. AB - A calmodulin-dependent protein phosphatase has been identified in human platelets by its cross-reactivity with an antibody developed against a bovine brain calmodulin-dependent protein phosphatase and by its calmodulin-stimulated dephosphorylation of 32P-labeled substrates. The platelet enzyme was partially purified to separate it from calmodulin and calmodulin-independent phosphatases. The partially purified enzyme was stimulated by calmodulin, requiring 15 nM calmodulin for half-maximal activation. Calmodulin increased the Vmax of the phosphatase, with no significant effect on its Km. The enzyme was stimulated irreversibly and made calmodulin-independent by limited proteolysis. The optimal pH for the phosphatase was 7.5. After partial purification, phosphatase activity was significantly increased in the presence of Mn2+ and Ca2+ over that observed in the presence of Ca2+ alone. The enzyme effectively dephosphorylated casein, histone, protamine, and platelet actin. The holophosphatase was estimated to have a molecular weight of 76,900 as determined by sedimentation on sucrose gradients. Immunoblotting techniques using an antibody against the brain phosphatase suggests that the enzyme consists of 2 subunits of 60,000 and 16,500 daltons; the 60,000-dalton subunit co-migrates in sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a 60,000-dalton calmodulin-binding protein in the platelet suggesting that it is the calmodulin-binding subunit of the enzyme. The identification of a calmodulin-dependent protein phosphatase in human platelets suggests a role for Ca2+-dependent dephosphorylation in platelet activation. PMID- 2987268 TI - Human serum amyloid P component. cDNA isolation, complete sequence of pre-serum amyloid P component, and localization of the gene to chromosome 1. AB - Complementary DNA clones corresponding to the human serum amyloid P component (SAP) were isolated, and the complete nucleotide and derived amino acid sequence of preSAP was determined. PreSAP biosynthesis is directed by a 1.1-kilobase mRNA. Synthesis and postsynthetic processing of preSAP in Xenopus oocytes result in secretion of a protein with mobility similar to native purified SAP when analyzed by sodium dodecyl sulfate gel electrophoresis. The human SAP gene is on chromosome 1, probably closely linked to the gene for C-reactive protein which encodes the related acute phase reactant found in human plasma. PMID- 2987269 TI - B-Z DNA junctions contain few, if any, nonpaired bases at physiological superhelical densities. AB - The reactions of bromoacetaldehyde (BAA) with recombinant plasmids that contain sequences which can adopt left-handed Z structures or, at other locations, cruciforms were studied as a function of supercoil density. The sequence in pRW756 that undergoes a supercoil induced transition from a right to left-handed helix was (dC-dG)16 and regions near the replication origin of the pBR322 vector were converted from linearforms to cruciforms. The locations of the most nonpaired structural features were mapped by S1 nuclease cleavage of the "wedged open" duplexes after linearization of the DNAs. Three cruciforms in the pBR322 portions of the plasmids were specifically detected by BAA reaction at physiological supercoil densities (sigma = -0.067). However, the B-Z junctions did not react with BAA under these conditions although the junctions were present since the (dC-dG)16 was shown to be left-handed. Thus, the B-Z junctions have less single-stranded character than the pBR322 cruciforms (3-6 nonpaired bases) and may be fully paired. At much higher superhelical densities (sigma = -0.11 0.12), the B-Z junctions as well as the cruciforms react with BAA indicating a change in the nature of the junctions. Studies were also performed with pRW777 which harbors the mouse kappa immunoglobin sequence (dT-dG)32 . (dC-dA)32 that adopts a left-handed helix under appropriate conditions; the results were similar to those found with pRW756. PMID- 2987270 TI - The neurovirulence of human and animal rotaviruses in cercopithecus monkeys. AB - Cercopithecus monkeys were inoculated according to the specifications of neurovirulence safety test for live rubella virus vaccine with RIT 4237, a rotavirus vaccine candidate. RIT 4237 is a high passage level of the Nebraska Calf Diarrhoea Virus (NCDV). The histological findings in the first test indicated some involvement of the central nervous system. The same test was therefore repeated with RIT 4237, with a lower passage level of the NCDV strain, and with the 'Wa' strain, a human virus grown in tissue culture. Clinical signs and histological findings were concordant and demonstrated that all the viruses were moderately neurovirulent. As in the poliovirus neurovirulence test, the histological lesions depended mainly upon a correct inoculation in the lumbar cord. RIT 4237 was found to have the same degree of neurovirulence as the low passage NCDV or as the 'Wa' strain. PMID- 2987271 TI - Neuropathic ulcers of the foot. AB - We report a prospective study of the causes and treatment of 26 long-standing neuropathic ulcers of the foot in 21 patients. The most important causal factor, well illustrated by pressure studies, was the presence of a dynamic or static deformity leading to local areas of peak pressure on insensitive skin. All but one of the 26 ulcers had healed after an average of 10 weeks of treatment in a light, skin-tight plaster cast, with the prohibition of weight-bearing. Recurrent ulceration was prevented in all but one foot by early operation to correct the causative deformity; this was performed after the ulcer had healed and before allowing weight-bearing on the limb. Pressure studies after operation confirmed that pressure points had been relieved. PMID- 2987272 TI - Direct nerve stimulation for painful peripheral neuropathies. AB - Nineteen patients with chronic pain due to a traumatic peripheral neuropathy were treated by means of implanted nerve stimulation. In 11 (58%) pain was completely relieved and in four (21%) it was reduced sufficiently to discontinue analgesics. The average follow-up was 11.5 months. The technique is described and the failures discussed. The necessity for implanting the stimulator proximally is emphasised. PMID- 2987273 TI - Permeabilization of rat hepatocytes with Staphylococcus aureus alpha-toxin. AB - Pathogenic staphylococci secrete a number of exotoxins, including alpha-toxin. alpha-Toxin induces lysis of erythrocytes and liposomes when its 3S protein monomers associate with the lipid bilayer and form a hexomeric transmembrane channel 3 nm in diameter. We have used alpha-toxin to render rat hepatocytes 93 100% permeable to trypan blue with a lactate dehydrogenase leakage less than or equal to 22%. Treatment conditions included incubation for 5-10 min at 37 degrees C and pH 7.0 with an alpha-toxin concentration of 4-35 human hemolytic U/ml and a cell concentration of 13-21 mg dry wt/ml. Scanning electron microscopy revealed signs of swelling in the treated hepatocytes, but there were no large lesions or gross damage to the cell surface. Transmission electron microscopy indicated that the nucleus, mitochondria, and cytoplasm were similar in control and treated cells and both had large regions of well-defined lamellar rough endoplasmic reticulum. Comparisons of the mannose-6-phosphatase and glucose-6-phosphatase activities demonstrated that 5-10 U/ml alpha-toxin rendered cells freely permeable to glucose-6-phosphate, while substantially preserving the selective permeability of the membranes of the endoplasmic reticulum and the functionality of the glucose-6-phosphatase system. Thus, alpha-toxin appears to have significant potential as a means to induce selective permeability to small ions. It should make possible the study of a variety of cellular functions in situ. PMID- 2987274 TI - The small subunit of ribonucleotide reductase is encoded by one of the most abundant translationally regulated maternal RNAs in clam and sea urchin eggs. AB - In both clam oocytes and sea urchin eggs, fertilization triggers the synthesis of a set of proteins specified by stored maternal mRNAs. One of the most abundant of these (p41) has a molecular weight of 41,000. This paper describes the identification of p41 as the small subunit of ribonucleotide reductase, the enzyme that provides the precursors necessary for DNA synthesis. This identification is based mainly on the amino acid sequence deduced from cDNA clones corresponding to p41, which shows homology with a gene in Herpes Simplex virus that is thought to encode the small subunit of viral ribonucleotide reductase. Comparison with the B2 (small) subunit of Escherichia coli ribonucleotide reductase also shows striking homology in certain conserved regions of the molecule. However, our attention was originally drawn to protein p41 because it was specifically retained by an affinity column bearing the monoclonal antibody YL 1/2, which reacts with alpha-tubulin (Kilmartin, J. V., B. Wright, and C. Milstein, 1982, J. Cell Biol., 93:576-582). The finding that this antibody inhibits the activity of sea urchin embryo ribonucleotide reductase confirmed the identity of p41 as the small subunit. The unexpected binding of the small subunit of ribonucleotide reductase can be accounted for by its carboxy terminal sequence, which matches the specificity requirements of YL 1/2 as determined by Wehland et al. (Wehland, J., H. C. Schroeder, and K. Weber, 1984, EMBO [Eur. Mol. Biol. Organ.] J., 3:1295-1300). Unlike the small subunit, there is no sign of synthesis of a corresponding large subunit of ribonucleotide reductase after fertilization. Since most enzymes of this type require two subunits for activity, we suspect that the unfertilized oocytes contain a stockpile of large subunits ready for combination with newly made small subunits. Thus, synthesis of the small subunit of ribonucleotide reductase represents a very clear example of the developmental regulation of enzyme activity by control of gene expression at the level of translation. PMID- 2987275 TI - Formyl peptide chemotaxis receptors on the rat neutrophil: experimental evidence for negative cooperativity. AB - To examine the existence of negative cooperativity among formyl peptide chemotaxis receptors, steady-state binding of f Met-Leu-[3H]Phe to viable rat neutrophils and their purified plasma membranes was measured and the data were subjected to statistical analysis and to computer curve fitting using the NONLIN computer program. Curvilinear, concave upward Scatchard plots were obtained. NONLIN and statistical analysis of the binding data indicated that a two saturable-sites model was preferable to a one-saturable-site model and statistically valid by the F-test (P less than .010). In addition, Hill coefficients of 0.80 +/- 0.02 were obtained. Kinetic dissociation experiments using purified plasma membranes showed evidence of site-site interactions of the destabilizing type (negative cooperativity). Thus, unlabeled f Met-Leu-Phe accelerated the dissociation of f Met-Leu-[3H]Phe under conditions where no rebinding of radioligand occurred. The rate of dissociation of f Met-Leu-[3H]Phe from the plasma membranes was dependent on the fold excess of unlabeled f Met-Leu Phe used in the dilution medium; at the highest concentration tested (10,000-fold excess), the dissociation rate was more than double the dissociation rate seen with dilution alone. In addition, occupancy-dependent affinity was ascertained directly by studying the effect of increasing fractional receptor saturation with labeled ligand on the dissociation rate of the receptor-bound labeled ligand. These data showed that the f Met-Leu-[3H]Phe dissociation rate was dependent on the degree of binding site occupancy over the entire biologically relevant range of formyl peptide concentrations. Furthermore, monitoring of the time course of dissociation of the receptor/f Met-Leu-[3H]Phe receptor/f Met-Leu-[3H]Phe complex as a function of receptor occupancy revealed that receptor affinity for f Met-Leu Phe remained occupancy-dependent during the entire time of dissociation examined (up to 10 min). Finally, the average affinity profile of the equilibrium binding data demonstrated a 60% decrease in receptor affinity in changing from the high affinity to the low affinity conformation. PMID- 2987276 TI - Transferrin receptor induction is required for human B-lymphocyte activation but not for immunoglobulin secretion. AB - Transferrin receptors are expressed on proliferating cells and are required for their growth. Transferrin receptors can be detected after, but not before, mitogenic stimulation of normal peripheral blood T and B cells. In the experiments reported here we have examined the regulation of transferrin receptor expression on activated human B cells and whether or not these receptors are necessary for activation to occur. Activation was assessed by studying both proliferation and immunoglobulin secretion. We have determined that transferrin receptor expression on B cells is regulated by a factor contained in supernatants of mitogen-stimulated T cells (probably B-cell growth factor). This expression is required for proliferation to occur, since antibody to transferrin receptor (42/6) blocks B-cell proliferation. Induction of immunoglobulin secretion, however, although dependent on PHA-treated T-cell supernatant, is not dependent on transferrin receptor expression and can occur in mitogen-stimulated cells whose proliferation has been blocked by antitransferrin receptor antibody. In addition, we have demonstrated that IgM messenger RNA induction following mitogen stimulation is unaffected by antitransferrin receptor antibody. These findings support a model for B-cell activation in which mitogen (or antigen) delivers two concurrent but distinct signals to B cells: one, dependent on B-cell growth factor and transferrin receptor expression, for proliferation, and a second, dependent on T cell-derived factors and not requiring transferrin receptors, which leads to immunoglobulin secretion. PMID- 2987277 TI - Regulation of discoidin I gene expression in dictyostelium discoideum by cell cell contact and cAMP. AB - We have previously presented evidence that cell-cell contact is the normal developmental signal to deactivate discoidin I gene expression in D discoideum [Berger EA, Clark JM: Proc Natl Acad Sci USA 80:4983, 1983]. Here we provide genetic evidence to support this hypothesis by examining gene expression in a cohesion-defective mutant, strain EB-21, which enters the developmental program but is blocked at the loose mound stage. When this strain was developed in suspension, the cells remained almost entirely as single amoebae, unlike the wild type, which formed large multicellular aggregates. In both strains, discoidin I mRNA levels were low in vegetative cells but rose sharply during the first few hours of development. However, the peak level reached at 8 hr in EB-21 exceeded that observed in wild type, and while the level declined markedly over the next few hours in wild type, it remained highly elevated in the mutant. Thus, there was a correlation between the inability of EB-21 to form normal cell-cell contacts and its deficiency in inactivating discoidin I gene expression. Previous studies from several laboratories, including this one, have demonstrated that exogenously added cAMP can block or reverse the changes in gene expression normally seen upon cell disaggregation. This has led us to propose that cAMP serves as a second messenger regulating the expression of contact-regulated genes. Here we provide additional support for this hypothesis. Intracellular cAMP levels rapidly dropped several-fold when wild type tight cell aggregates were disaggregated and remained low as the cells were cultured in the disaggregated state. Furthermore, overexpression of discoidin I mRNA late in development in EB 21 was corrected by addition of high concentrations of cAMP. These results are consistent with a second messenger function for cAMP in the contact-mediated regulatory response, and they indicate that the cAMP response machinery for discoidin I gene expression is capable of functioning in the cohesion-defective EB-21 strain. PMID- 2987278 TI - A beta-type transforming growth factor, present in conditioned cell culture medium independent of cell transformation, may derive from serum. AB - An alpha-type transforming growth factor (TGF alpha) is produced at high levels by rat embryo cells transformed by the Snyder-Theilen strain of feline sarcoma virus (FeSV). Addition of 2 ng mouse epidermal growth factor (mEGF) during purification identified the presence of a second, EGF-dependent growth factor of the TGF beta type (TGF beta) in this conditioned medium. This factor had an approximate Mr of 12,000 and eluted at 37% acetonitrile during high performance liquid chromatography. This extracellular type of TGF beta activity also was present in conditioned medium of rat cells after infection with a transformation defective strain of Abelson leukemia virus, and hence expression of this growth factor activity was independent of cell transformation. Moreover, the presence of an EGF-dependent, 12,000 Mr clonogenic activity in extracts of bovine serum alone suggests serum as an origin for the B-type transforming growth factor initially observed in conditioned medium of Snyder-Theilen FeSV transformed cells. This does not, however, preclude the possibility that TGF beta is also secreted by the transformed rat embryo cells themselves. PMID- 2987279 TI - Silica-bound polyethyleneglycol as stationary phase for separation of proteins by high-performance liquid chromatography. PMID- 2987280 TI - High-performance liquid chromatographic analysis of basic drugs on silica columns using non-aqueous ionic eluents. I. Factors influencing retention, peak shape and detector response. AB - The use of silica columns together with non-aqueous ionic eluents provides a stable yet flexible system for the high-performance liquid chromatographic analysis of basic drugs. At constant ionic strength, eluent pH influences retention via ionisation of surface silanols and protonation of basic analytes, pKa values indicating the pH of maximum retention. At constant pH, retention is proportional to the reciprocal of the eluent ionic strength for fully protonated analytes and quaternary ammonium compounds. The addition of water up to 10% (v/v) has little effect on retention if the protonation of the analytes is unaffected. Thus, it is likely that retention is mediated primarily via cation exchange with surface silanols. However, additional factors must play a part with compounds such as morphine which give tailing peaks at acidic or neutral eluent pHs. PMID- 2987281 TI - Detection of urinary amine metabolites in toluene diisocyanate exposed rats. PMID- 2987282 TI - Simplified procedure for the quantitation of radioactive phosphoinositides by thin-layer chromatography. PMID- 2987283 TI - Rapid large-scale isolation of biologically active molecules using reversed-phase "flash" chromatography: initial purification of endogenous Na+, K+-ATPase inhibitors from human urine. PMID- 2987284 TI - Development and preliminary evaluation of two ELISAs for detection of anti-CMV Ig and IgM antibodies. PMID- 2987286 TI - Adenosine 3',5'-monophosphate levels in human follicular fluid: relationship to oocyte maturation and achievement of pregnancy after in vitro fertilization. AB - cAMP, estradiol (E2), and progesterone levels were determined in 24 follicular fluid samples obtained from 8 women who conceived after in vitro fertilization and in 47 samples from 26 women who did not. Follicular development was induced by human menopausal gonadotropin, and maturation of retrieved oocytes was assessed by the degree of cumulus mucification and corona dispersal. The mean follicular fluid cAMP concentration was significantly (P less than 0.001) lower in women who became pregnant than in those who did not (106 vs. 241 pmol/ml), while the mean E2 level was significantly (P less than 0.01) higher (727 vs. 497 ng/ml), and the progesterone to E2 ratio was significantly (P less than 0.05) lower (9.5 vs. 18.0). Overall, follicles of immature, intermediate, and mature oocytes did not differ in cAMP content. However, intermediate and mature oocytes from women who became pregnant were derived from follicles containing significantly (P less than 0.01) lower cAMP levels than those of women who did not become pregnant (66 and 122 vs. 233 and 288 pmol/ml, respectively). Furthermore, fertilized oocytes leading to conception originated from follicles with significantly (P less than 0.001) lower cAMP concentrations than the follicles that yielded nonfertilized oocytes or fertilized oocytes not leading to conception (92 vs. 270 and 240 pmol/ml, respectively). Similarly, significantly (P less than 0.05) lower cAMP levels were found in the follicular fluid of cleaved oocytes resulting in a pregnancy compared to those that did not (86 vs. 236 pmol/ml). It is concluded that low levels of cAMP are associated with successful fertilization and cleavage of human oocytes in vitro resulting in viable pregnancies and may, therefore, be used as a marker of optimal follicular development in in vitro fertilization cycles. PMID- 2987285 TI - Effects of continued adrenocorticotropin stimulation on the mineralocorticoid hormones in classical and nonclassical simple virilizing types of 21-hydroxylase deficiency. AB - Superphysiological doses of ACTH were administered for 3 consecutive days to nine patients with CAH, five with the classical simple virilizing (CSV) type and four with nonclassical simple virilizing type (NCSV; late onset), receiving a sodium restricted diet. In the CSV patients, cortisol levels were lower [4.9 +/- 2.2 (+/ SEM) micrograms/dl] than in the NCSV patients (10.9 +/- 3.8; P less than 0.005) and normal subjects (10.7 +/- 4.0; P less than 0.05). ACTH produced a subnormal increase to only 14.5 micrograms/dl by day 3. In the NCSV patients, cortisol rose slowly during the first 24 h, but reached normal response levels by 48 h (42.5 +/ 11.5 micrograms/dl). In all patients, basal plasma corticosterone and 18 hydroxydeoxycorticosterone (18-OHDOC) levels were normal, but deoxycorticosterone (DOC) was elevated at 25.3 +/- 5.0 ng/dl (P less than 0.05). ACTH failed to increase plasma levels of DOC, corticosterone, and 18-OHDOC. Aldosterone and 18 hydroxycortisol were elevated in both groups [29.1 +/- 5.8 (P less than 0.02) and 83.8 +/- 15.3 (P less than 0.01) ng/dl, respectively] and increased briskly after the first 24 h of ACTH. However, neither steroid returned to normal levels in the CSV group, but both did in the NCSV group. Paired values of stimulated cortisol and aldosterone in normal subjects and CSV and NCSV patients (n = 76) were significantly negatively correlated (r = -0.63; P less than 0.001), suggesting that cortisol inhibits aldosterone biosynthesis. Prolonged ACTH administration after initial increases returned aldosterone and 18-hydroxycortisol levels from the zona glomerulosa to baseline values in the NCSV type, but not in the CSV type. The capacity to increase cortisol levels, which occurred only in NCSV patients, is linked to the reduction of aldosterone in the zona glomerulosa. In contrast, in both types of 21-hydroxylase deficiency, sustained impairment of both the 17-hydroxy pathway (cortisol) and the 17-deoxy pathway of the zona fasciculata (DOC, corticosterone, and 18-OHDOC) was demonstrated. PMID- 2987287 TI - A factor produced by cultured rat Leydig tumor (Rice 500) cells associated with humoral hypercalcemia stimulates adenosine 3',5'-monophosphate production via the parathyroid hormone receptor in human skin fibroblasts. AB - The Rice-500 Leydig cell tumor of Fischer rats is associated with humoral hypercalcemia in vivo and produces a factor that stimulates cAMP formation in cultured rat osteosarcoma cells. We found that cultured human skin fibroblasts respond to both human PTH-(1-34) and the factor produced by cultured rat Leydig tumor cells with a dose-dependent rise in cAMP formation. The time courses for stimulation of the two agents were similar, and stimulation by both was blocked by the competitive PTH antagonist [8,18-norleucine,34-tyrosine]bovine PTH-(3-34) amide. These data suggest that PTH-like factors secreted by a murine tumor are capable of interacting with the human PTH receptor. PMID- 2987288 TI - Increased cyclic nucleotide phosphodiesterase (PDE) and calmodulin activities in soluble fraction of Graves' thyroid: analysis of increase in Ca+2 dependence of PDE activities. AB - Phosphodiesterase (PDE) and calmodulin (CaM) activities were studied in soluble fractions of normal and Graves' thyroid tissue. Normal and Graves' thyroid tissues were obtained at thyroid surgery. PDE activities were assayed with cAMP and cGMP as substrates. CaM activity was measured as the ability to activate bovine thyroid CaM-dependent PDE. cAMP and cGMP PDE activities were increased 1.5 and 2.2-fold above normal in Graves' thyroid, respectively. The major cause of the increase in enzyme activities was their higher Ca+2 dependence. CaM activity also was 1.6-fold increased in Graves' thyroid, although it probably does not contribute to the increase in the Ca+2 dependence of PDE activities because of its relative sufficiency vs. PDE in normal thyroid. Three forms of cAMP and cGMP PDE activities were eluted from Sephadex G-200 columns, with mol wt of 280,000, 140,000, and 80,000-100,000, respectively. The first peak had little or no CaM dependence, the second peak had moderate or dominant CaM dependence, and the third peak revealed weak or dominant CaM dependence. The increase in the CaM dependent form of the third peak of PDE activity in Graves' thyroid tissue may explain the increase in Ca+2 dependence of PDE activities. PMID- 2987289 TI - In vitro modulation of prolactin binding to human mammary carcinoma cells by steroid hormones and prolactin. AB - This study demonstrates the specific binding of human (h) PRL to mammary carcinoma cells of the newly established line EFM-19. Under saturating conditions, [125I]hPRL bound to these cells with high affinity (Ka = 4.3 X 10(10) M-1) and low capacity (4080 binding sites/cell). In serum-free medium, stimulation of cell proliferation by PRL was found, suggesting a biological role in the growth of human breast cancer. hPRL was more effective in binding to and promoting the growth of EFM-19 cells than ovine PRL or other lactogenic hormones. Up- and down-regulation of [125I]hPRL binding occurred after pretreatment of EFM 19 cell cultures with subphysiological or higher hPRL concentrations, respectively. Physiological concentrations of 17 beta-estradiol or dihydrotestosterone increased the cellular capacity of hPRL binding. Pharmacological concentrations of dihydrotestosterone or physiological concentrations of progesterone reduced the binding of [125I]hPRL. The results provide evidence for complex regulatory mechanisms of PRL binding by human mammary carcinoma cells involving steroid hormones. PMID- 2987290 TI - Erythrocyte sodium/potassium adenosine triphosphatase in thyroid disease and nonthyroidal illness. AB - Thyroid hormone is known to modulate cell membrane sodium/potassium adenosine triphosphatase (Na/K-ATPase). To determine whether the activity of this enzyme differed in patients with nonthyroidal illness with low levels of circulating thyroid hormones and patients with documented clinical hypothyroidism, we measured Na/K-ATPase activity in red blood cells from patients with hypo- and hyperthyroidism, patients with nonthyroid disease with and without reduced circulating levels of thyroid hormone, and normal subjects. We also assessed whether the activity of this enzyme reflects decreased thyroid hormone action at the cellular level in patients with nonthyroidal illness. Hyperthyroidism was associated with decreased and hypothyroidism with increased erythrocyte Na/K ATPase activity [142 +/- 24 (+/- SE) and 371 +/- 37 nmol Pi/mg X h; P less than 0.05 and P less than 0.01 compared to normal]. Enzyme activity in cells from patients with nonthyroidal illness and low levels of circulating T3 was significantly higher than that in cells from normal subjects (289 +/- 11 vs. 223 +/- 16 nmol Pi/mg X h; P less than 0.01), but was not significantly different from that in cells from hypothyroid patients. Red cell Na/K-ATPase activity in patients with nonthyroidal illness and normal thyroid function tests (185 +/- 38 nmol Pi/mg X h was indistinguishable from normal values. These data confirm that hyperthyroid patients have decreased red cell Na/K-ATPase activity and provide direct evidence that erythrocyte ATPase activity is increased in hypothyroid patients. The change in enzyme activity in patients with nonthyroidal illness and decreased circulating T3 levels was comparable to that in hypothyroidism. These results suggest that since red cell Na/K-ATPase activity does not distinguish between ill patients with low thyroid function tests and those with hypothyroidism, tissue hypothyroidism may exist in the former group of patients. PMID- 2987291 TI - Euthyroid hyperthyroxinemia and thyroxine-binding prealbumin excess in islet cell carcinoma. AB - This manuscript describes euthyroid hyperthyroxinemia secondary to elevated serum T4-binding prealbumin (TBPA) concentrations in a patient with islet cell carcinoma and reports serum TBPA measurements in other patients with islet cell carcinoma. A 73-yr-old man with a 17-yr history of metastatic islet cell carcinoma was found to have hyperthyroxinemia. His total serum T4 concentration was 18.5 micrograms/dl (normal, 5.5-11.5). Eight years previously, his serum T4 concentration was normal. His free T4 concentration, as determined by equilibrium dialysis, was 1.3 ng/dl (normal, 0.9-2.1). Serum T3, TSH, and T4-binding globulin (TBG) concentrations were normal, as was the TSH response to TRH administration. Polyacrylamide gel electrophoresis of the patient's serum in the presence of tracer amounts of [125I]T4 revealed that, compared to normal sera, [125I]T4 binding to TBPA was increased from 30.0 +/- 6.0% (mean +/- SD) to 52.0%. The distribution of [125I]T3 amont albumin, TBG, and TBPA was normal in this electrophoretic procedure. The concentration of TBPA in the patient's serum was 189 mg/dl. In contrast, the mean serum TBPA concentration in normal men was 40 +/ 4 (mean +/- SD) mg/dl, and that in normal women was 18 +/- 4 mg/dl. Sera from the patient's daughter, his brother, 2 sisters, and 2 male paternal cousins contained normal amounts of TBPA, ranging from 34-47 mg/dl in his male relatives and from 26-30 mg/dl in his female relatives. Serum free T4 index determinations in his relatives also were normal. Serum TBPA concentrations were determined in 14 additional patients with islet cell carcinoma. In 1 of these patients, a man with an insulinoma, serum TBPA was elevated (66 mg/dl). This patient's serum T4 level was 7.3 micrograms/dl, and his free T4 index was 7.0. These data and another study suggest that islet cell carcinoma may rarely produce a TBPA-like protein resulting in an elevated serum TBPA concentration. Markedly elevated serum TBPA is associated with euthyroid hyperthyroxinemia. PMID- 2987292 TI - Prevalence and clinical associations of intraocular pressure changes in Graves' disease. AB - We investigated an unselected series of 55 patients with treated or untreated hyperthyroid Graves' disease, assessing their clinical and laboratory status and ophthalmological findings, including the difference in intraocular pressure (dIOP) between upgaze and straight gaze using applanation tonometry. An increased dIOP (greater than 2 mm Hg) was detected in only 22% of Graves' patients [who had a mean dIOP of 3.5 +/- 1.6 (+/- SEM) mm Hg]. dIOP did not correlate with age, sex, age at disease onset, duration of disease, mode of antithyroid treatment, or thyroid function testing at the time of examination. Mean Hertel exophthalmometry measurements in patients with a dIOP greater than 2 mm Hg were 22.0 +/- 2.9 mm compared with 18.4 +/- 3.7 mm in those with a dIOP less than 2 mm Hg (P less than 0.027, by Wilcoxon rank sum test). Only 58% of patients with increased dIOP had clinical exophthalmos, but all had other evidence of Graves' eye disease. Computed tomographic scanning revealed significant proptosis and/or orbital muscle involvement in all of the patients with increased dIOP. PMID- 2987294 TI - Pituitary-adrenal responsiveness to corticotropin-releasing hormone in patients receiving chronic, alternate day glucocorticoid therapy. AB - We examined the responsiveness of the pituitary-adrenal axis to ovine corticotropin-releasing hormone (oCRH) in 14 women with systemic lupus erythematosus receiving chronic, alternate day glucocorticoid therapy with prednisone. Testing was done twice and in a random order (at 2000 h) on the day when the steroid was taken (12 h after the last dose) and on the day when no glucocorticoid was administered (36 h after the last dose). Plasma ACTH and cortisol responses were markedly blunted on the day of treatment and mildly blunted on the day off treatment compared to those in normal subjects. Altered metabolic clearance of exogenous oCRF was not responsible for this difference, since the plasma disappearance curves of immunoreactive oCRH were similar on both days. The degree of suppression was dependent on the dose of prednisone, and the amount of cortisol secreted during the oCRH test was directly proportional to the logarithm of the concurrent plasma ACTH level. Thus, the cortisol response to ACTH was normal in all patients. These data suggest that the blunting of responsiveness to oCRH on both days of testing represents prednisolone suppression of the corticotroph cell. Despite this, the adrenal glands retain normal responsiveness to ACTH, suggesting that moderate decreases in daily ACTH secretion are compatible with sustaining normal adrenal function. Hence, the site of the mild suppression of the hypothalamic-pituitary-adrenal axis during chronic, alternate day treatment with glucocorticoids is central, whereas the adrenal glands appear to remain functionally unaffected. PMID- 2987293 TI - Effect of bromocriptine treatment on the aldosterone response to angiotensin II and adrenocorticotropin in idiopathic hyperaldosteronism. AB - Bromocriptine can prevent an increase in plasma aldosterone during the infusion of angiotensin II in normal subjects and during upright posture in some patients with idiopathic hyperaldosteronism. To determine if bromocriptine prevents the increase in plasma aldosterone concentration during angiotensin II infusion in idiopathic hyperaldosteronism, we infused angiotensin II in five patients with idiopathic hyperaldosteronism, before and after treatment with bromocriptine (2.5 mg, three times daily for 5 days), and measured the resulting plasma aldosterone and angiotensin II concentrations. We also determined the adrenal response to ACTH infusion before and after bromocriptine treatment in four of these patients. Bromocriptine treatment did not significantly change the plasma concentrations of aldosterone before or during the infusions of angiotensin II and ACTH. It did significantly decrease mean blood pressure and increase the plasma corticosteroid concentrations in the preinfusion periods, but it did not alter the response of blood pressure to angiotensin II or of plasma corticosteroid concentrations to the ACTH infusions. PMID- 2987295 TI - The human vasoactive intestinal peptide receptor: molecular identification by covalent cross-linking in colonic epithelium. AB - To characterize the molecular components of the vasoactive intestinal peptide (VIP) receptor in human intestine, [125I]VIP was covalently bound to human colonic epithelial membranes using dithio-bis(succinimidyl propionate). Sodium dodecyl sulfate-polyacrylamide gel autoradiographic studies of affinity labeled membranes revealed three major bands corresponding to [125I]VIP-protein complexes of 66,000, 33,000, and 16,000 mol wt. Labeling of the 66,000 and 33,000 mol wt complexes was specific, since it was abolished by VIP, while labeling of the 16,000 mol wt complex was not. Densitometric scanning of autoradiographs indicated that labeling of the 66,000 mol wt complex was inhibited by low VIP concentrations in the 10(-10)-10(-8) M range, but was unaffected by glucagon or octa-cholecystokinin. It was also reduced by VIP-(2-28), with a potency 1/100th that of VIP, and by GTP in the concentration range of 10(-7)-10(-3) M. The 33,000 mol wt complex behaved similarly to the 66,000 mol wt complex with respect to specificity and GTP sensitivity, but differed in one major feature, its affinity for VIP. Its labeling was inhibited by native VIP concentrations in the 10(-9) 10(-7) M range. Assuming one molecule of [125I]VIP bound per molecule of protein, two proteins of 63,000 and 30,000 mol wt were identified as VIP-binding sites. The 63,000 mol wt protein had the properties expected for the VIP receptor coupled to adenylate cyclase in human colon, while the 30,000 mol wt protein was a low affinity binding site. Treatment of human colonic membranes with the sulfhydryl reducing agent dithiothreitol before [125I]VIP binding strongly reduced the labeling of the two proteins. This finding does not support the hypothesis that the low affinity 30,000 mol wt binding site may be a monomer of the high affinity binding site. PMID- 2987296 TI - Studies of renin activation and regulation of aldosterone and 18 hydroxycorticosterone biosynthesis in hyporeninemic hypoaldosteronism. AB - In an attempt to evaluate deficiencies of renin activation and adrenal zona glomerulosa biosynthesis in hyporeninemic hypoaldosteronism (HH), we studied active and inactive renin (AR and IR, respectively) responses to the dopamine antagonist metoclopramide, furosemide, and graded dose infusion of ACTH in 10 HH patients and 6 normal subjects. In HH patients, AR levels, but not IR levels, were decreased relative to normal values. While normal subjects had an AR response to metoclopramide, the HH patients did not. The AR response to furosemide in HH patients was markedly diminished compared to that in normal subjects. Plasma cortisol and corticosterone levels were in the normal range, but the zona glomerulosa products 18-hydroxycorticosterone (18-OHB) and aldosterone (Aldo) were low in HH patients. Plasma 18-OHB and Aldo responses to metoclopramide and furosemide were diminished, but cortisol and 18-OHB responses to ACTH were normal in the HH patients. Our observation that 18-OHB and Aldo responses to metoclopramide were diminished refutes the possibility that excessive adrenal zona glomerulosa dopaminergic activity could account for reduced biosynthesis of 18-OHB and Aldo in HH patients. Our results appear most consistent with the concept that the primary etiological factor in the HH syndrome is impairment of renal activation of renin. PMID- 2987297 TI - Influence of the preventive effect of eicosapentaenoic acid on cerebrovascular disorders. PMID- 2987298 TI - Effect of sodium polyanetholesulfonate and gelatin on the recovery of Gardnerella vaginalis from blood culture media. AB - Sodium polyanetholesulfonate (SPS) is used as a routine supplement to blood culture media to enhance recovery of microorganisms, but it inhibits the growth of Peptostreptococcus anaerobius, Neisseria meningitidis, Neisseria gonorrhoeae, and Streptobacillus moniliformis. Comparative clinical blood culture studies at the University of Colorado Hospital suggested that SPS also inhibits the growth of Gardnerella vaginalis. We inoculated 16 blood culture isolates of G. vaginalis into 11 blood culture media containing SPS or sodium amylosulfate, with and without gelatin. In the absence of gelatin, only brain heart infusion and thiol broths with SPS supported the growth of more than five strains of G. vaginalis, whereas all media except Bactec 6B and 7C and brucella broths recovered most isolates with SPS and gelatin or with sodium amylosulfate alone. We conclude that SPS inhibits the growth of G. vaginalis in blood culture media but that this inhibition is medium dependent and can be overcome by supplementation of most media with gelatin. PMID- 2987299 TI - Rapid subtyping of equine herpesvirus 1 with monoclonal antibodies. AB - Two antigenically similar subtypes of equine herpesvirus 1 (EHV-1) cause disease in horses. A procedure for rapid differentiation of the two EHV-1 subtypes with monoclonal antibodies was developed. Subtype-specific pools of monoclonal antibodies were constructed, characterized, and used in enzyme immunofiltration and indirect immunofluorescence assays to subtype 50 epizootiologically unrelated field isolates of EHV-1. Both assays allowed accurate subtype identification of each EHV-1 isolate with the monoclonal antibody pools. The subtyping procedures were simple and amenable to typing many isolates at one time and permitted unambiguous EHV-1 subtype identification with 3 h after isolation of the virus in tissue culture. PMID- 2987300 TI - Epidemiology of rotavirus electropherotypes determined by a simplified diagnostic technique with RNA analysis. AB - The incidence and RNA electropherotypes of rotavirus in stools or rectal swabs of children with diarrhea were studied for three rotavirus seasons (1981 through 1984) in Philadelphia, Pa. We used a simplified RNA analysis method involving polyacrylamide gel electrophoresis followed by silver staining. Phosphate buffered saline suspensions of the stools and swab eluates were examined directly by polyacrylamide gel electrophoresis-silver staining analysis and enzyme-linked immunoadsorbent assay (Rotazyme; Abbott Laboratories); electron microscopy was performed on solid stool specimens. The RNA analysis results were compared with electron microscopy and enzyme-linked immunosorbent assay results and exhibited a sensitivity and specificity greater than or equal to that of electron microscopy or the enzyme-linked immunosorbent assay. Ten different electropherotypes were detected among the 68 rotavirus RNA-positive specimens examined over the 3-year study. The predominant electropherotype was different in each season. Our results indicate that the polyacrylamide gel electrophoresis-silver nitrate strain RNA analysis of simple unextracted stool suspensions is a uniquely useful diagnostic technique; it rapidly provides both a definitive positive result and immediate determination of the RNA electropherotype, which is of value for epidemiological study. PMID- 2987301 TI - Rapid herpes simplex virus detection in clinical samples submitted to a state virology laboratory. AB - Of 16,779 specimens received for herpes simplex virus (HSV) isolation since 1982, 4,465 (26.6%) were positive for HSV by either standard tissue culture or an antigen detection system (peroxidase-antiperoxidase; PAP). The overall isolation rate for genital vesicle specimens was lower (26.1%) than that for nongenital specimens (29.3%). Monthly isolation rates ranged from 19 to 32% for genital specimens and from 20 to 44% for nongenital specimens. Increasing demands for HSV isolation led to comparison of tissue culture with PAP. In the first comparison, HSV was isolated in single human fibroblast cell cultures from 1,019 of 4,261 specimens (23.9%), whereas single human fibroblast wells stained at 24 and 72 h postinoculation were PAP positive for 1,007 of 4,261 specimens (23.6%). In the second comparison, HSV was isolated from 225 of 1,026 (21.9%) specimens and duplicate human foreskin fibroblast cell wells stained at 24 and 72 h were PAP positive in 241 of 1,026 (23.5%). With the dual-well PAP system, all results were reported within 72 h, approximately 70% of positives were reported within 24 h, and considerable savings in time and materials resulted. PMID- 2987302 TI - Comparison of serodiagnosis of group B coxsackievirus infections by an immunoglobulin M capture enzyme immunoassay versus microneutralization. AB - A solid-phase enzyme immunoassay (EIA) with the capture antibody principle was developed and evaluated for detection of immunoglobulin M (IgM) specific for group B coxsackieviruses (CBV) in human serum. Of 19 patients who were culture positive for CBV type 4 infections, the EIA for CBV type 4 IgM was more sensitive (84%; 16/19 positive) than a standard microneutralization test (16%; 3/19 positive) and a microneutralization assay on the IgM fractions obtained by sucrose density gradient fractionation of human serum (68%; 13/19 positive). The attempt to produce a serotype-specific EIA was not successful, as IgM antibodies were detected to heterotypic CBV. The pooled CBV types 2 to 5 IgM EIA demonstrated an overall sensitivity of 88% (21/24 positive) in comparison to virus isolation for CBV types 4 and 5. The EIA was specific for IgM to CBV, with the exception of some cross-reactions with hepatitis A IgM-positive sera (15%; 3/20). In comparison to the microneutralization test, the EIA for pooled CBV IgM was more rapid, less cumbersome, and more sensitive. Commercially available reagents used in this study enable the CBV IgM EIA to be developed in diagnostic virology laboratories. PMID- 2987303 TI - In vitro detection of porcine rotavirus-like virus (group B rotavirus) and its antibody. AB - Four isolates of porcine rotavirus-like virus (PRVLV) infected MA104 cells and induced syncytium formation after low-speed centrifugation of the inoculum onto the monolayer. Ten of 44 (23%) Ohio swine sera had PRVLV antibodies when tested by indirect immunofluorescence, using PRVLV-infected MA104 cell monolayers as antigen. PMID- 2987304 TI - Metastatic skeletal disease in the pediatric population. AB - We reviewed the autopsy, clinical, and radiographic records of 39 patients with metastatic skeletal disease (age range 18 months-20 years). There were 11 different primary tumors responsible for skeletal metastases, of which neuroblastoma was the most common (16 patients). Other commonly occurring tumors giving rise to secondary skeletal deposits were rhabdomyosarcoma (seven), teratoma-teratocarcinoma (four), and Wilms tumor (three). Overall patient survival ranged from 2 weeks to 72 months, with an average of 10.2 months. Those patients presenting without skeletal metastases at the time of diagnosis of the primary tumor had a survival time averaging 16.2 months (range 3-72 months), whereas those patients presenting initially with skeletal metastases at the time of diagnosis of the primary tumor had a survival time of 8.5 months (range 0.5-23 months). PMID- 2987306 TI - Modulation of procollagen gene expression by retinoids. Inhibition of collagen production by retinoic acid accompanied by reduced type I procollagen messenger ribonucleic acid levels in human skin fibroblast cultures. AB - Recent clinical observations have suggested that retinoids, which are in frequent use in dermatology, can affect the connective tissue metabolism in skin and other tissues. In this study, we examined the effects of several retinoids on the metabolism of collagen by human skin fibroblasts in culture. Incubation of cultured fibroblasts with all-trans-retinoic acid or 13-cis-retinoic acid, in 10( 5) M or higher concentrations, markedly reduced the procollagen production, as measured by synthesis of radioactive hydroxyproline. The effect was selective in that little, if any, inhibition was noted in the incorporation of [3H]leucine into the noncollagenous proteins, when the cells were incubated with the retinoids in 10(-5) M concentration. Similar reduction in procollagen production was noted with retinol and retinal, whereas an aromatic analogue of retinoic acid ethyl ester (RO-10-9359) resulted in a slight increase in procollagen production in these cultures. The reduction in procollagen production by all-trans-retinoic acid was accompanied by a similar reduction in pro alpha 2(I) of type I procollagen specific messenger RNA (mRNA), as detected by dot blot and Northern blot hybridizations. Hybridizations with human fibronectin and beta-actin specific DNA probes indicated that the levels of the corresponding mRNAs were not affected by the retinoids, further suggesting selectivity in the inhibition of procollagen gene expression. Further control experiments indicated that all-trans retinoic acid, under the culture conditions employed, did not affect the posttranslational hydroxylation of prolyl residues, the mannosylation of newly synthesized procollagen, the specific radioactivity of the intracellular prolyltransfer RNA pool, or DNA replication. All-trans-retinoic acid also elicited a reduction in trypsin-activatable collagenase, but not in the activity of prolyl hydroxylase or an elastaselike neutral protease in the fibroblast cultures. Incubation of three fibroblast lines established from human keloids with all-trans-retinoic acid or 13-cis-retinoic acid also resulted in a marked reduction in procollagen production. The results, therefore, suggest that further development of retinoids might provide a novel means of modulating collagen gene expression in patients with various diseases affecting the connective tissues. PMID- 2987307 TI - Generation of free radical intermediates from foreign compounds by neutrophil derived oxidants. AB - A large number of foreign compounds, including many drugs, industrial pollutants, and environmental chemicals, can be oxidized under appropriate conditions to potentially toxic free radical intermediates. We evaluated the ability of the oxidants produced by the neutrophil myeloperoxidase system to generate free radical intermediates from several such compounds. Sodium hypochlorite or hypochlorous acid produced by human peripheral blood neutrophils and trapped in the form of taurine chloramine were both found to be capable of producing free radicals from chlorpromazine, aminopyrine, and phenylhydrazine. These radical intermediates were demonstrated by visible light spectroscopy and by direct electron spin resonance (for the chlorpromazine and aminopyrine radicals) or by spin-trapping (for the phenyl radical generated from phenylhydrazine). Stable oxidants produced by the neutrophils (i.e., those present in the supernatants of stimulated neutrophils in the absence of added taurine) also were found to be capable of generating free radical intermediates. The production of the oxidants and the ability of neutrophil supernatants to generate these radicals were almost completely eliminated by sodium azide, a myeloperoxidase inhibitor. We suggest that the oxidation by neutrophils of certain chemical compounds to potentially damaging electrophilic free radical forms may represent a new metabolic pathway for these substances and could be important in the processes of drug toxicity and chemical carcinogenesis. PMID- 2987308 TI - Diarrhea in streptozocin-treated rats. Loss of adrenergic regulation of intestinal fluid and electrolyte transport. AB - Diarrhea was noted in rats with streptozocin-induced chronic diabetes. We have investigated the possibility that this diarrhea is a consequence of altered neuronal control of water and electrolyte absorption in the intestinal epithelium. In particular, we examined noradrenergic control because alpha-2 adrenergic agonists are known to stimulate intestinal fluid absorption. When compared with nondiabetic littermates, chronically diabetic rats exhibited significant impairment of fluid absorption by the ileum and colon, but not the jejunum. This impairment of intestinal fluid absorption was not found in either insulin-treated or untreated acutely diabetic (7 d) animals. Mucosal histology appeared normal in all of the above groups. Mucosal norepinephrine stores in the jejunum and ileum of chronically diabetic rats were estimated in vitro by the short-circuit current (Isc) response to tyramine, an agent that effectively releases stored norepinephrine. Pargyline was added to inhibit enzymatic destruction of the added tyramine. In chronically diabetic rats, the Isc response to tyramine was significantly decreased in ileum, but not in jejunum. However, when these responses were expressed as a fraction of the maximal Isc tissue response to exogenously added epinephrine, significant decreases were noted in both ileum and jejunum. In tissues from acutely diabetic rats, Isc responses to tyramine and epinephrine were no different from controls. When sympathetic denervation was produced in nondiabetic rats by treatment with 6-OH-dopamine, the pattern of impaired fluid absorption that developed was the same as that observed in chronically diabetic rats. We conclude that impaired intestinal mucosal absorption of fluid and electrolytes slowly develops in rats made diabetic with streptozocin and that this absorptive impairment is due to a loss of normally present noradrenergic innervation of enterocytes. PMID- 2987305 TI - Relationship of urinary and blood carbon dioxide tension during hypercapnia in the rat. Its significance in the evaluation of collecting duct hydrogen ion secretion. AB - This study was designed to establish the relationship between urinary pCO2 and systemic blood pCO2 during acute hypercapnia and to investigate the significance of this relationship to collecting duct hydrogen ion (H+) secretion when the urine is acid and when it is highly alkaline. In rats excreting a highly alkaline urine, an acute increase in blood pCO2 (from 42 +/- 0.8 to 87 +/- 0.8 mmHg) resulted in a significant fall in urine minus blood (U-B) pCO2 (from 31 +/- 2.0 to 16 +/- 4.2 mmHg, P less than 0.005), a finding which could be interpreted to indicate inhibition of collecting duct H+ secretion by hypercapnia. The urinary pCO2 of rats with hypercapnia, unlike that of normocapnic controls, was significantly lower than that of blood when the urine was acid (58 +/- 6.3 and 86 +/- 1.7 mmHg, P less than 0.001) and when it was alkalinized in the face of accelerated carbonic acid dehydration by infusion of carbonic anhydrase (78 +/- 2.7 and 87 +/- 1.8 mmHg, P less than 0.02). The finding of a urinary pCO2 lower than systemic blood pCO2 during hypercapnia suggested that the urine pCO2 prevailing before bicarbonate loading should be known and the blood pCO2 kept constant to evaluate collecting duct H+ secretion using the urinary pCO2 technique. In experiments performed under these conditions, sodium bicarbonate infusion resulted in an increment in urinary pCO2 (i.e., a delta pCO2) which was comparable in hypercapnic and normocapnic rats (40 +/- 7.2 and 42 +/- 4.6 mmHg, respectively) that were alkalemic (blood pH 7.53 +/- 0.02 and 7.69 +/- 0.01, respectively). The U-B pCO2, however, was again lower in hypercapnic than in normocapnic rats (15 +/- 4.0 and 39 +/- 2.5 mmHg, respectively, P less than 0.001). In hypercapnic rats in which blood pH during bicarbonate infusion was not allowed to become alkalemic (7.38 +/- 0.01), the delta pCO2 was higher than that of normocapnic rats which were alkalemic (70 +/- 5.6 and 42 +/- 4.6 mmHg, respectively, P less than 0.005) while the U-B pCO2 was about the same (39 +/- 3.7 and 39 +/- 2.5 mmHg). We further examined urine pCO2 generation by measuring the difference between the urine pCO2 of a highly alkaline urine not containing carbonic anhydrase and that of an equally alkaline urine containing this enzyme. Carbonic anhydrase infusion to hypercapnic rats that were not alkalemic resulted in a fall in urine pCO(2) (from 122+/-5.7 to 77+/-2.2 mmHg) which was greater (P <0.02) than that seen in alkalemic normocapnic controls (from 73+/- 1.9 to 43+/ 1.3 mmHg) with a comparable urine bicarbonate concentration and urine nonbicarbonate buffer capacity. CO(2) generation, therefore, from collecting dust H(+) secretion and titration of bicarbonate, was higher in hypercapnic rats that in normocapnic controls. We conclude that in rats with actue hypercapnia, the U-B p(CO(2)) achieved during bicarbonate loading greatly underestimates collecting duct H(+) secretion because it is artificially influenced by systemic blood pCO(2). the deltapCO(2) is a better qualitative index of collecting duct H+ secretion that the U-B pCO(2), because it is not artificially influenced by systemic blood pCO(2) and it takes into account the urine PCO(2) prevailing before bicarbonate loading. PMID- 2987309 TI - Use of SM-1 monoclonal antibody and human complement in selective killing of small cell carcinoma of the lung. AB - SM-1 is a murine monoclonal antibody strongly reactive with a cell membrane antigen of small cell carcinoma (SCC) of the lung but unreactive with the membrane of most other carcinomas and normal tissues including normal bone marrow. We have found that in the presence of human complement, SM-1 antibody is highly cytotoxic to SCC cells. Using three treatments with antibody and complement, more than 99% of SCC cells in culture were lysed, as determined by the chromium release and clonogenic assays. Similar efficiency of SCC cell lysis was observed when one SM-1 antibody treatment was followed by three treatments with human complement. In contrast, there was little antibody-dependent lysis of non-small cell lung cancer cells, other carcinomas, and leukemia cell lines. The amount of chromium released from normal bone marrow cells treated with SM-1 antibody and complement was minimal and was mainly due to the effect of complement alone. Clonogenic assays, including colony-forming unit granulocytic/monocytic, erythroid burst-forming unit, and colony-forming unit granulocytic/erythroid/monocytic/megakaryocytic, also showed no significant SM-1 antibody-dependent cytotoxicity on normal bone marrow precursors. Since SM-1 antibody is selectively cytotoxic to SCC cells in the presence of human complement, it is a potentially useful agent for the selective eradication of tumor cell contamination in marrows of patients with metastatic small cell lung cancer and possibly for in vivo serotherapy. PMID- 2987310 TI - Activation of the respiratory burst enzyme from human neutrophils in a cell-free system. Evidence for a soluble cofactor. AB - Activation of the respiratory burst in phagocytic cells, an important host defense process, is not yet well understood. We now report the development of a cell-free system for activation of NADPH oxidase, the respiratory burst enzyme, in human neutrophils. Activation was achieved by the addition of arachidonic acid to a postnuclear supernatant (500 g) from disrupted unstimulated cells (no arachidonate, 0.2; with arachidonate, 3.4 nmol superoxide anion/min per mg) and was dependent on both the concentration of arachidonate and on the amount of cellular material present. Activity stimulated by arachidonate appeared to be NADPH oxidase based on a Michaelis constant for NADPH of 32 microM and a pH optimum of 7.0-7.5. Separation of the 500-g supernatant by high speed centrifugation revealed a requirement for both soluble and particulate cofactors. Activation of NADPH oxidase by arachidonate did not occur in the high speed pellet fraction from unstimulated cells but could be restored by the addition of the high speed supernatant. In addition, priming of intact neutrophils with low concentrations of the chemoattractant N-formyl-methionyl-leucyl-phenylalanine or the tumor promoter phorbol myristate acetate replaced the soluble factor requirement for NADPH oxidase activation by arachidonate in the high speed pellet. This cell-free system can now be used to provide further insight into the biochemical basis of priming and the terminal mechanisms involved in the activation of NADPH oxidase. PMID- 2987312 TI - Mesoblastic nephroma contains fibronectin but lacks laminin. AB - Non-metastatic mesoblastic nephromas from four young children were shown to contain fibronectin but not laminin using an immunoperoxidase staining procedure. In contrast, one metastasising spindle celled sarcomatous tumour from a neonate was laminin positive. During embryogenesis primitive nephrogenic mesenchyme contains only fibronectin and no laminin; metanephric blastema (permanent kidney) is positive for laminin. The staining for fibronectin and laminin may help to ascertain the histogenesis of different types of renal tumour. PMID- 2987313 TI - Comparison of ELISA with virus isolation for the diagnosis of genital herpes. AB - An enzyme linked immunosorbent assay (ELISA) system which detects and simultaneously types herpes simplex virus antigens in clinical specimens from patients with genital herpes has been compared with standard tissue culture isolation. Although more sensitive than a similar method previously described and also more sensitive than electron microscopy and immunofluorescence, ELISA did not detect all the viruses isolated in tissue culture. Costs were comparable. The speed of obtaining the result together with knowledge of the type causing infection are useful when antiviral chemotherapy is envisaged and when considering the likelihood of recurrences. PMID- 2987314 TI - Biphenotypic leukaemia: a case of mixed T lymphoblastic and myeloblastic leukaemia. AB - A case of mixed acute leukaemia with T lymphoblastic, myeloblastic, and monocytic components is described. The use of immunological markers, ultrastructural morphology, cytochemistry, immunochemistry, and combined techniques, simultaneously detecting two markers in individual cells, made it possible to define the different blast cell populations. PMID- 2987315 TI - Localization of serotonin immunoreactivity in the opossum cerebellum. AB - We have used the indirect antibody peroxidase-antiperoxidase technique to analyze the course of serotonin (5-hydroxytryptamine; 5HT) fibers to the deep cerebellar nuclei; the distribution of serotonin within the nuclei; the continued course of 5HT fibers to the cerebellar cortex; and the lobular and laminar distribution of this indoleamine in the cerebellar cortex. Only rarely are fibers found in either the restiform body or the brachium pontis. However, a distinct bundle of serotoninergic axons is present in the medial aspect of the brachium conjunctivum. Axons arise from this bundle and course dorsally into the neuropil of the deep cerebellar nuclei. The densest immunostaining is present in posterior and ventral regions of all four cerebellar nuclei. Within the nuclei large (24% of total) and small (76% of total) varicosities are present. The average distance between varicosities on individual axons is 3.85 micron (S.D. = 1.2). The innervation of the cerebellar cortex is derived primarily from fibers that course through the deep nuclei. At levels caudal to the deep nuclei a single midsagittal band courses into lobules VIII and IX. In the cerebellar cortex, serotoninergic axons and varicosities are present in all lobules; however, the fiber density is not uniform. The densest distribution is present in vermal lobule VIII and the dorsal folia of lobule IX. Within the granule cell layer of lobules VIII and IX, immunoreactive elements form a midsagittal band, and to a lesser degree, two parasagittal bands. Beaded serotoninergic fibers course through the deep and middle portion of the granule cell layer and give rise to a plexus at the border between the Purkinje cell and granule cell layers. Within this plexus axons extend long distances in the transverse and sagittal planes. Long beaded axons oriented in the transverse plane of the folia are also present in the deep molecular layer. A few radial serotoninergic fibers ascend to the pial surface and give rise to very short tangential branches. In all three cortical layers, both large (19% of total) and small (81% of total) varicosities are present. The average distance between varicosities on individual fibers is 5.3 micron (S.D. = 2.2).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2987316 TI - Neuronal connections in the primary auditory cortex: an electrophysiological study in the cat. AB - Neuronal connections in the primary auditory cortex (AI) of the cat were studied electrophysiologically by using intracellular recording techniques. Fast conducting fibers from the medial geniculate nucleus (MG) projected monosynaptically onto AI neurons in layers III-VI (mainly in layer IV), whereas slow-conducting MG-fibers projected monosynaptically onto AI neurons in layer I. AI neurons which received monosynaptic inputs from the auditory association cortices (AII and Ep) and/or from the contralateral AI were distributed in all layers of the AI; the commissural fibers from the contralateral AI were divided into fast- and slow-conducting ones. AI neurons were categorized into seven types: type I neurons which received monosynaptic inputs from slow-conducting MG fibers were located in layer I. Type II neurons which received polysynaptic inputs from the MG were located in layers II-VI. Type III neurons which sent their axons to the AII or Ep were mainly located in layer III. Type IV neurons which sent their axons to the contralateral AI were located mainly in layer III. Type V neurons which received monosynaptic inputs from fast-conducting MG-fibers were located mainly in layer IV. Type VI neurons which projected onto the inferior colliculus were located in the upper part of the layer V. Type VII neurons which projected onto the MG were located in layers V and VI. PMID- 2987311 TI - Activation of human neutrophil nicotinamide adenine dinucleotide phosphate, reduced (triphosphopyridine nucleotide, reduced) oxidase by arachidonic acid in a cell-free system. AB - Sonicates from unstimulated human neutrophils produce no measurable superoxide since the superoxide-generating enzyme, NADPH oxidase, is inactive in these preparations. Previous attempts to activate the oxidase in disrupted cells with conventional neutrophil stimuli have been unsuccessful. This report describes a cell-free system in which arachidonic acid (82 microM) was able to activate superoxide generation that was dependent upon the presence of NADPH and the sonicate. For activation to occur, both the particulate and supernatant fractions of the sonicate must be present. Calcium ions, which are required for activation of intact neutrophils by arachidonate, were not necessary in the cell-free system. In quantitative terms, the superoxide-generating activity in the cell free system could account for at least 20-50% of the superoxide rate observed in intact neutrophils stimulated with arachidonate. Sonicates from patients with chronic granulomatous disease (CGD) could not be activated by arachidonic acid in the cell-free system. In three patients representing both genetic forms of CGD, the defect appeared to reside in the particulate fraction. The soluble cofactor was normal in all three patients and could be used to activate normal neutrophil pellets in the presence of arachidonic acid. Thus, at least a portion of the activation mechanism in the neutrophil, that residing in the soluble phase, appeared to be normal in patients with CGD. PMID- 2987317 TI - A reticular pattern of intrinsic connections in primate area V2 (area 18). AB - A system of periodic intrinsic connections is demonstrated in area V2 (area 18) of squirrel and macaque monkeys by large injections of tritiated amino acids, horseradish peroxidase (HRP), and fluorescent latex beads. These connections originate from pyramidal neurons concentrated in layers 3 and 5. Terminations occur in all cortical layers, largely coextensive with labeled neurons but more restricted in layer 4. This multilaminar distribution contrasts with the mainly supragranular localization of periodic intrinsic connections in V1 (area 17), and may imply a close interaction, in V2, of periodic intrinsic connections with pulvinocortical, as well as with corticocortical terminations (concentrated, respectively, in layers 3 and 5, and in lower 3 and 4). As in V1, the tangential configuration of these connections in V2 is reticular or latticelike, and is detectable for 2.5-3.0 mm from an injection site of HRP, 3H amino acids, or latex beads. Cross-sectional widths of labeled regions vary from 250 to 800 micron in squirrel monkey and from 400 to 1,000 micron in macaque, depending on which portion of the lattice is measured. When periodic intrinsic connections are compared with stripes labeled histochemically by cytochrome oxidase (CO), no clear relationship is obvious between the two systems. This result contrasts with the orderly tangential alignment reported between CO-reactive zones in V2 and certain extrinsic connections; namely, pulvinocortical terminations (Livingstone and Hubel, '82) and clusters of neurons projecting to area V4 (DeYoe and Van Essen, '84). Other extrinsic connections, however, such as backgoing connections from V2 to V1, do not seem to have a periodic distribution. Thus, although some discontinuous cortical connections relate to each other in a precise mosaic fashion, intrinsic and some extrinsic connections may observe different modes of organization. PMID- 2987319 TI - Aneurysmal (angiomatoid) fibrous histiocytoma. AB - We report a patient with aneurysmal (angiomatoid) fibrous histiocytoma manifested clinically as a single firm subcutaneous nodule, diagnosed as an epithelial cyst. Histologically, the growth showed massive histiocytic proliferation and areas with a vascular component. The lesion was histologically mistaken for Kaposi's sarcoma. PMID- 2987318 TI - A comparative immunoperoxidase and histopathologic study of condylomata acuminata. AB - Thirty-seven lesions from 29 patients with condylomata acuminata were comparatively studied by peroxidase-antiperoxidase (PAP) technique and routine histopathology. PAP staining demonstrated papillomavirus common antigen (PCA) mostly in the granular layer of the epidermis in 22 of 37 lesions (59%) of condylomata acuminata. Presence of PCA is closely correlated with the following histopathologic findings: 1) coarse keratohyalin granules; 2) perinuclear vacuolization or koilocytotic cells in the granular layer; 3) hyperkeratosis with focal parakeratosis. From these findings it is concluded that these histopathologic findings suggest the presence of PCA, and that the presence of papillomavirus may play a role in the formation of these histopathologic features of condylomata acuminata. PMID- 2987320 TI - Cholera toxin, cyclic AMP, and the firefly flash. AB - Cholera toxin, injected into living fireflies, caused a delayed, non-hormone dependent activation of adenylate cyclase, an elevation of cyclic AMP levels, and a brilliant persistent glow of the firefly light organ 8-20 hours following injection. During periods of spontaneous flashing, onset of toxin-induced glowing was abrupt and step-like in character, due to a markedly prolonged off-time of individual flashes. These observations provide a striking demonstration of the mechanism of cholera toxin action, and, together with other data, suggest that the initiation of the normal adult firefly flash is mediated through elevation of cyclic AMP levels secondary to the activation of an octopamine-sensitive adenylate cyclase. PMID- 2987321 TI - Effects of thyroidectomy, insulin, and phospholipids on cyclic AMP phosphodiesterase in rat adipocyte plasma membranes. AB - The effect of thyroid status on the cyclic nucleotide phosphodiesterase in adipocyte plasma membranes has been studied. In euthyroid rat fat cells, about 7% of the total cAMP phosphodiesterase was found in the plasma membrane. Thyroidectomy doubled both the enzyme's total activity in this fraction, and its specific activity (60 versus 37 pmol/min/mg) compared to euthyroid rat plasma membrane. In membranes from thyroidectomized rats, phosphodiesterase hydrolyzed cAMP with a single Km of 2 microM, whereas in euthyroid rat membranes, Lineweaver Burk plots were non-linear, with apparent Kms of 0.5 and 5 microM. This phosphodiesterase activity was insensitive to exogenous guanine nucleotides and calcium. In vivo injection of triiodothyronine restored phosphodiesterase activity in plasma membranes from thyroidectomized rats to the values obtained for euthyroid rats. Centrifugation on a 10 to 45% sucrose density gradient of the plasma membrane fractions gave two main peaks of phosphodiesterase activity which hydrolyzed the cAMP in adipocyte plasma membranes from both euthyroid and thyroidectomized rats. The distribution profiles for these activities were very similar in the two plasma membrane preparations. The peaks of phosphodiesterase and 5'nucleotidase activity coincided. Thyroidectomy raised the phosphodiesterase activity of these two peaks, particularly of the first. The cAMP phosphodiesterase activities in both hypothyroid and control plasma membrane preparations were also sensitive to insulin and were activated by phospholipase A2 and three anionic phospholipids. Thyroid hormones therefore regulate the degradation of cAMP in plasma membranes by a mechanism which seems different from the one involved in the action of insulin, and is independent of the membrane phospholipid composition. PMID- 2987322 TI - Production of monoclonal cyclic GMP and cyclic AMP antisera. AB - In order to facilitate the in situ localization of cyclic nucleotides, a large supply of high-titer anti-sera for immunohistologic analyses would be valuable. Therefore, monoclonal antibodies (Mab) directed at 2'-0-succinyl cyclic GMP (s cGMP) or 2'-0-succinyl cyclic AMP (s-cAMP) were generated. Immunized mice developed polyclonal antibodies of high titer to either cyclic nucleotide as reflected in an enzyme-linked immunosorbent assay (ELISA) employed to monitor these anti-sera. With the use of standard hybridoma technology and ELISA to monitor anti-cyclic nucleotide antibody production from positive clones, several Mab-producing clones were grown to large volume in tissue culture and ascites fluid. The anti-cGMP and anti-cAMP anti-sera, which demonstrated significant ELISA titers at greater than 1:2,000,000 dilutions, are monospecific and can be employed for radioimmunoassays, ELISA, or immunohistologic localization of cyclic nucleotides in situ. PMID- 2987324 TI - Effects of interpersonal problem-solving training with chronic aftercare patients on problem-solving component skills and effectiveness of solutions. PMID- 2987323 TI - Aspects of intracellular transit of serum and lipid phases of milk. PMID- 2987325 TI - Ultrastructural study of epithelial cell attachment to implant materials. AB - Attachment of gingival tissues to the protruding post of an endosseous dental implant is of great importance for the prognosis of its clinical success. Little is known about the requirements an implant material must meet to secure a durable permucosal seal. The objective of this study, therefore, was to gain insight into the morphology of epithelial cell-implant interfaces for a number of well-known or potential implant materials. Guinea pig epithelial cells were cultured on gold, titanium, carbon, hydroxylapatite, carbonate apatite, and modified polystyrene substrates. The metallic and carbon substrates were obtained by vapor phase deposition of thin films on polystyrene carriers. This technique allowed for the preparation of ultra-thin sections for transmission electron microscopy containing substrate as well as the cells adhering to it. The cultures on apatite were decalcified prior to sectioning. The results revealed tht as attachment structures, focal contacts, extracellular matrix contacts, and hemidesmosome-like contracts could be distinguished. Hemidesmosome-like contacts were only observed on apatite and polystyrene and not on the metallic or carbon surfaces. The results of this study suggest the existence of some, as yet unknown, property of the substrate that determines the nature and the structure of the contact site with epithelial cells. PMID- 2987326 TI - Erythema multiforme and herpes simplex virus. AB - It has been suggested that herpes simplex virus (HSV) can trigger erythema multiforme (EM) at different times. One recent study showed HSV antigen in immune complexes of patients with EM. The purpose of our study was to assess a possible association between EM and HSV. Sixteen patients and 16 matched controls were studied using an enzyme-linked immunosorbent assay (ELISA) to measure antibody of the IgG, IgA, and IgM classes against HSV-1. From our study on patients with oral erythema multiforme, we found no evidence to correlate the occurrence and/or severity of EM and the HSV-1. PMID- 2987327 TI - Photo-oxidation of tetracycline adsorbed on hydroxyapatite in relation to the light-induced staining of teeth. AB - In an attempt to understand the chemistry of the light-induced staining of children's teeth by tetracycline, we studied the photo-chemical behavior of tetracycline adsorbed on hydroxyapatite, as a simple model of enamel. Tetracycline was strongly bound by hydroxyapatite to give a pale yellow material which, under ultraviolet light, showed a bright yellow fluorescence (lambda max 525 nm). On exposure of this material to the radiation from a medium-pressure mercury lamp, the fluorescence gradually disappeared, and a red-purple product was formed. Photo-acoustic spectroscopy was employed to follow the disappearance of tetracycline and the concomitant formation of the red-purple product, the spectrum of which (lambda max 530 nm) led to the conclusion that it was 4 alpha, 12 alpha-anhydro-4-oxo-4-dedimethylaminotetracycline (AODTC). This assignment was supported by the observation that 1 mole of oxygen was absorbed per mole of adsorbed tetracycline converted to the red-purple product. It is suggested that the formation of AODTC on hydroxyapatite probably takes place by a mechanism of photo-oxidation similar to that already proposed for solutions of tetracycline, and that the formation of AODTC in children's teeth is responsible for the light induced staining caused by tetracycline. PMID- 2987328 TI - Measured levels of antibody to the herpes simplex virus in patients with oral cancer and controls. PMID- 2987329 TI - The availability of fluoride chemical supplies. AB - The availability of adequate supplies of fluoride chemicals is a critical factor in consistently maintaining the optimal fluoride concentration in water supply systems. The most recent shortage of fluoride chemicals occurred in 1982. The factors precipitating and contributing to periodic fluoride chemical shortages are discussed and recommendations for preventing future shortages are outlined. PMID- 2987330 TI - Products of myocardial ischemia and electrical instability of the heart. AB - Products of ischemic metabolism can make a major contribution to electrical instability of the heart. In particular, enhanced extracellular potassium slows conduction, thereby predisposing to reentrant circuits, especially when levels of cyclic adenosine monophosphate (AMP) increase in the early ischemic period. Other products (lactate, lipid metabolites) alter the duration of the action potential. Accumulated calcium may induce inward diastolic currents. PMID- 2987332 TI - Studies on the mechanism of toxicity of the mycotoxin sporidesmin. IV. Inhibition by copper-chelating agents of the generation of superoxide radical by sporidesmin. AB - Sporidesmin, the mycotoxin responsible for "facial eczema" in ruminants, has previously been shown to generate superoxide free radical, the latter being formed during autoxidation of the reduced form of the mycotoxin, a dithiol. The autoxidation of reduced sporidesmin is catalysed by iron and by copper, although cobalt, nickel, manganese, cerium, vanadium and molybdenum were found to be without effect. The catalytic activity of copper was some 900-fold that of iron. Cyanide and certain organic complexing agents inhibit superoxide production from reduced sporidesmin by virtue of their ability to chelate copper. Some proteins behave likewise, again through binding of the metal catalyst, although amino acid bound copper was found to be as effective as ionic copper in catalysing the autoxidation reaction. From the results of the present experiments, it is suggested that any superoxide production from sporidesmin in vivo would be mediated by the intracellular transport pool of copper. Furthermore, the ability of zinc to inhibit intestinal absorption of copper may be involved in the mechanism whereby salts of this metal afford protection against the harmful effects of sporidesmin in the living animal. PMID- 2987331 TI - The heart as a sensory organ. AB - Three groups of receptors in the heart are activated by changes in pressure in the cardiac chambers. Those at the venous-atrial junctions with myelinated vagal afferent nerves indicate changes in heart rate and degree of atrial filling. A second group, present in all the cardiac chambers, served by unmyelinated vagal afferent nerves, signals changes in ventricular preload, afterload and cardiac contractility. A third group, also present in all the cardiac chambers, has both myelinated and unmyelinated afferent nerves that pass to the spinal cord. Their normal function is unknown. Abnormal activation of the cardiac mechanoreceptors during myocardial ischemia may be important in the genesis of life-threatening arrhythmias. PMID- 2987333 TI - Effects of morphine on somatocardiac sympathetic reflexes in spinalized cats. AB - The effects of morphine on sympathetic reflexes, recorded in the inferior cardiac nerve, to myelinated A and unmyelinated C afferent stimulation were tested in 17 acutely spinalized cats. Stable sympathetic A and C reflexes of short latency (approximately 30 ms and 140 ms in the case of the ulnar nerve, respectively) could be recorded in the inferior cardiac sympathetic nerve to stimulation of somatic A and C afferents in the ulnar and upper thoracic intercostal nerves, ipsilaterally. Spinal sympathetic A reflexes, which were primarily evoked from stimulation of A delta afferent fibers, could be elicited from more segmental levels than could sympathetic C reflexes. Additionally, smaller reflexes, only from A afferent fiber activation, were identified from stimulations on the contralateral side of the body. Small doses of morphine (0.02 mg kg-1, i.v.) proved to be ineffective at altering sympathetic A and C reflexes, while somewhat larger doses (0.2 mg kg-1, i.v.) produced a clear 62% decrease in C reflexes and a 33% decrease in A reflexes, Dosages of 1 and 2 mg kg-1 severely depressed both A and C reflexes. All of the above effects of morphine administration were completely and immediately reversible by naloxone (i.v.). The results are discussed with regard to the effects of morphine on sympathetic A and C reflexes in CNS intact, anesthetized cats. PMID- 2987334 TI - Central nervous system opiate and 5-hydroxytryptamine influences on baroreflex control of the coronary circulation. AB - The effect of intravenous infusion of fentanyl (an opiate receptor agonist, 0.55 microgram kg-1 min-1) on the control of the circumflex coronary circulation was examined in unsedated dogs at rest and during baroreceptor stimulation evoked by acute rises in aortic pressure (balloon inflation in thoracic aorta). Circumflex flow was measured using Doppler flow transducers in dogs with experimental complete heart block and with ventricles paced at a constant rate. Studies were also performed before and one week after intracisternal injection of the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), to examine the role of CNS 5 hydroxytryptamine (5-HT) in any sympathetic vasoconstrictive effects. Fentanyl infusion caused after a few minutes a progressive rise in resting aortic pressure and a significant fall in circumflex conductance; circumflex flow usually fell. Atrial rate also fell. The gain of the baroreflex control of circumflex conductance was enhanced by fentanyl. One week after intracisternal 5,7-DHT, the gain of the baroreflex in each dog was diminished. When fentanyl was infused into these preparations, no consistent changes in resting atrial rate, aortic pressure and circumflex conductance could be observed, but all dogs showed a recovery of the coronary baroreflex gain towards values observed before intracisternal 5,7 DHT. These data suggest that the gain control of coronary baroreflexes is influenced by CNS opiate and 5-HT dependent mechanisms. PMID- 2987336 TI - Effect of carbon dioxide on the activity of slowly and rapidly adapting pulmonary stretch receptors in cats. AB - Single fiber activity from slowly and rapidly adapting pulmonary stretch receptors (PSR) was recorded from the left cervical vagus of anesthetized, open chested and artificially ventilated cats. Reducing the end-tidal CO2 to low values did not affect the frequency of occurrence of higher threshold (HT) PSR. Occlusion of the left pulmonary artery (LPA) had no significant effect on the resting discharge of both HT and low threshold (LT) PSR. Cyclic ventilation with 8% CO2 in O2 reduced the activity of LT and HT receptors by similar amounts, irrespective of their anatomical location. After LPA occlusion, CO2 ventilation reduced markedly the activity of both type of PSR but to the greatest extent that of the HT receptors, the majority of which were located in the intrapulmonary airways. The CO2 depressant effect may not be due solely to changes in H+ concentration at the receptor level, since acetazolamide did not totally abolish the effect even though it significantly reduced it. Sustained inflation with 8% CO2 in O2 significantly reduced the activity of HT receptors in both the dynamic and static phases of inflation, but had no effect on the activity of LT receptors. Direct localization showed that the receptors which were more accessible to CO2 (all HT and one LT) were located in the lung parenchyma. In the case of rapidly adapting receptors, sustained inflations with CO2 gave inconsistent results. The results show clearly that, as in other mammalian species, the PSR activity in cats is also reduced by hypercapnia. The present study stresses the importance of localizing the PSR and making the observations separately on the two types of PSR, for there may be qualitative and quantitative differences. PMID- 2987335 TI - Split medulla preparation in the cat: arterial chemoreceptor reflex and respiratory modulation of the renal sympathetic nerve activity. AB - The study was undertaken in order to assess the changes in sympathetic output in a split medulla preparation of the cat which, as shown earlier, has impaired respiratory rhythm generation. The effects of medullary midsagittal sections on renal sympathetic nerve firing were investigated in chloralose anesthetized, paralyzed and artificially ventilated cats. Recordings of phrenic and recurrent laryngeal nerve activity served as indices of central respiratory rhythm generation. Sections, 5 mm deep from the dorsal medullary surface and extending 6 mm rostrally and 3 mm caudally to the obex, did not produce any significant changes in heart rate, blood pressure or tonic renal sympathetic nerve firing levels. They decreased or abolished, however, the respiratory rhythmicity in renal sympathetic nerve which paralleled the reduction of inspiratory discharges in phrenic and recurrent laryngeal nerves, and abolished the carotid body chemoreceptor-sympathetic reflex. The inspiratory activity remaining after the sections could still be enhanced by chemoreceptor stimulation. The inhibitory baroreceptor and pulmonary stretch receptor sympathetic reflexes, and the central excitatory effect of CO2 on renal sympathetic nerve firing were not altered. The effects of electrical stimulation within the midsagittal plane of the medulla have shown that descending pathways from the medullary inspiratory neurons (or their medullary collaterals) do not participate in the facilitation of spinal preganglionic neurons during inspiration and in relaying the pulmonary stretch receptor inhibitory sympathetic reflex. A region located close to the obex was identified from which excitatory responses in renal sympathetic nerves, compatible with the response obtained by carotid sinus nerve stimulation, could be evoked. It is concluded that a lesion in the midline of the lower medulla at the level of the obex selectively destroys cells or pathways which relay the carotid body chemoreceptor-sympathetic reflex. PMID- 2987337 TI - Autonomic dysfunction and Eaton Lambert syndrome. AB - Autonomic dysfunction associated with Eaton Lambert syndrome in a patient with oat cell carcinoma of the lung is reported. Widespread postganglionic adrenergic and cholinergic defects suggested the presence of autonomic neuropathy. Clinical remission accompanied tumor regression after combined chemotherapy and radiation. PMID- 2987338 TI - Comparative study of conjugation between horseradish peroxidase and D-cytochrome b5. Incorporation into subcellular membranes. AB - Horseradish peroxidase was conjugated to D-cytochrome b5 by three different two step methods. The yield of conjugates based on the peroxidase enzymatic activity recovered after gel filtration was very low in the glutaraldehyde method, but higher in the N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) and periodate methods. The molecular size of the conjugates was analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. Monomeric conjugates were mostly formed via the glutaraldehyde and SPDP methods in the presence of appropriate molar ratios of proteins. Most of the conjugates formed via the periodate method were polymers. The conjugate preparations of the three methods could be incorporated into microsomal membranes. Conjugate polymers, however, appeared less able to be incorporated then monomers. There was a nonpreferential incorporation of free or conjugated D-cytochrome b5 contained in the conjugate preparation of the glutaraldehyde method. In conclusion, this study gives preference to the glutaraldehyde method for the preparation of conjugates that will subsequently be used as an in vivo marker of the D-cytochrome b5 incorporation into membranes. PMID- 2987339 TI - beta-NADPHase- and TMPase-positive "snake-like tubules" in the exocrine pancreas: cytochemical and immunocytochemical studies. AB - Using five different protocols, two enzymes, nicotinamide adenine dinucleotide phosphate phosphohydrolase (beta-NADPHase) and sodium trimetaphosphatase (TMPase), were localized in the acinar cell of rat pancreas by ultrastructural cytochemistry. The beta-NADPHase cytochemical localization was realized at pH 4.8 and pH 3.9. At pH 4.8, the beta-NADPHase activity was found in the Golgi intermediate saccules, lysosomes, gland lumen, and tubular structures, described as snake-like tubules (Beaudoin AR, Grondin G, Lord A: Eur J Cell Biol 33:275, 1984; Beaudoin AR, Grondin G, Lord A, Pelletier M: In Proc 42nd Ann Meeting Electron Microscopy Soc Am. San Francisco Press, CA, 1984). There was no detectable beta-NADPHase activity at pH 3.9. The TMPase cytochemistry was done at pH 3.9 according to Oliver (J Histochem Cytochem 28:78, 1980) and at pH 3.9 and 4.8 with the medium described by Berg (J Histochem Cytochem 8:92, 1960). TMPase localization varied according to the protocols. It was found in tubular structures described as "basal lysosomes," lysosomes, and zymogen granules, whereas Golgi saccules were generally negative. Our observations showed that the structures identified as "basal elongated lysosomes" and revealed by TMPase (Oliver C: J Histochem Cytochem 28:78, 1980; J Histochem Cytochem 31:1209, 1983) were morphologically similar to snake-like tubules (SLT) revealed by beta NADPHase. Relationships between SLT and mitochondria as well as lysosomes and plasma membranes were observed. Using amylase-specific antibodies, it was also shown, by the protein A-gold immunocytochemical technique, that SLT do not contain amylase and, in fasting conditions, would not be involved in the transport of secretory proteins. PMID- 2987340 TI - Intrarenal beta-receptor and renal baroreceptor interaction in the control of the renin response to transient reduction of the renal perfusion pressure in man. AB - We have examined the mechanisms mediating the release of renin elicited in man by reduction of renal perfusion pressure. Fifteen patients with essential hypertension and six normotensive subjects were investigated during diagnostic renal arteriography. Renal neural receptors were inhibited by propranolol (10 mg i.v.) and activated by a standard cold pressor test. Vascular receptors were stimulated by unilateral reduction of renal perfusion pressure by 50%, using a balloon-tipped catheter. The stimulus caused release of renin. In hypertensives, arterial plasma renin increased by 44, 69 and 73% of control at 5, 15 and 30 min, respectively. Adrenergic activation by cold raised the arterial and the renal venous renin by approximately 50% of control and caused a fourfold rise when it was combined with the arterial obstruction. Following propranolol the renin response to reduction of the renal perfusion pressure was delayed and reduced, and cold stimulation, both alone and in combination with arterial obstruction, failed to stimulate renin release. Findings were qualitatively and quantitatively similar in the normotensive group. This study supports the hypothesis that the renin response to reduction of renal perfusion pressure in man results from an interaction of adrenergic and vascular receptors. It cannot be stated whether the former are synergistic or supplementary to the latter, even though adrenergic activation by cold stimulation provides evidence that a synergism between the two may exist. PMID- 2987341 TI - Acute and chronic effects of the converting enzyme inhibitors enalapril and lisinopril on reflex control of heart rate in normotensive man. AB - The effects of enalapril maleate and its lysine analogue, lisinopril, on reflex control of heart rate after acute and chronic administration were examined in 10 normotensive males. Both drugs reduced blood pressure without change in heart rate after acute dosing and after seven days. Both drugs impaired the vagally mediated early cardiac acceleration associated with lying down (standing to lying test). A similar effect was observed following edrophonium. After lisinopril bradycardia induced by facial immersion (diving reflex) was significantly attenuated. Edrophonium similarly attenuated the bradycardia. Both these reflexes are parasympathetically mediated. Neither drug altered heart rate or blood pressure changes following Valsalva's manoeuvre and the cold pressor test. Plasma noradrenaline was unchanged. Absence of reflex tachycardia with blood pressure reduction by converting enzyme inhibitors may be related to increased parasympathetic activity either centrally or peripherally without impairment of baroreflexes or sympathetic function. PMID- 2987342 TI - Participation of the renin-angiotensin system in the maintenance of blood pressure during changes in posture in patients with essential hypertension. AB - To assess whether and how the activation of the renin-angiotensin system that occurs in response to changes in posture contributes to the maintenance of blood pressure, we measured blood pressure, heart rate, plasma noradrenaline and plasma active and inactive renin in patients with essential hypertension in the supine, sitting and standing positions, (each sustained for 30 min), before and after administration of captopril. These studies were performed before and after a brief period of diuretic therapy. Both in the normal and in the sodium depleted state captopril reduced the normal adaptive response of blood pressure to changes in body position, whereas the responses of heart rate and plasma noradrenaline to these stimuli were unaffected by captopril. In contrast, the postural increases in active renin were magnified after captopril while inactive renin was unchanged throughout these acute studies. Our results indicate that during mild but rather prolonged postural stresses the generation of angiotensin by active renin supports blood pressure either through its direct vasoconstrictive effect and/or by potentiating the activity of the sympathetic nervous system. It is unlikely that the changes in active renin depend upon conversion from the inactive form. PMID- 2987343 TI - Phenotypic expression of B lymphocytes. III. Marginal zone B cells in the spleen are characterized by the expression of Tac and alkaline phosphatase. AB - The marginal zone of the spleen contains lymphocytes with an intermediate morphologic form between small lymphocytes and plasmablasts. The majority of cells can be stained by B cell monoclonal antibodies including B1, Leu-14, and Leu-12. The most significant finding is the expression of an interleukin 2 receptor (Tac) and an enzyme-alkaline phosphatase by the marginal zone B lymphocytes. The Tac antigen is not normally present in B cells, but can be found in a portion of in vitro-activated B lymphocytes. In conjunction with other evidence, the marginal zone B lymphocytes may represent the activated B cells in the early stages of B cells differentiation. Alternatively, the Tac-positive, marginal zone B lymphocytes may be a discrete subpopulation of activated B cells. PMID- 2987344 TI - Characterization of a membrane surface glycoprotein associated with T-cell activation. AB - A cell surface antigen (gp140) was previously shown to exist on T cell subsets as well as on monocytes and macrophages in normal peripheral blood. Elevated expression of this antigen was associated with immune system disorders, acute lymphocytic leukemias, and in vitro activation of T cells. The antigen could be identified with monoclonal antibody (MAb) T305. Gp140 was a biosynthetic product of T cells because it could be labeled with [3H]leucine or [3H] glucosamine. Biochemical studies of gp140 used high performance liquid chromatography with nitrocellulose blotting to isolate aliquots suitable for 125I radiolabeling and immunoprecipitation to demonstrate: a) a reduction in m.w. of gp140 KD to 90 KD after deglycosylation by trifluoromethanesulfonic acid, b) alteration of isoelectric point from 4.1 to 5.7 after neuraminidase treatments, c) absence of N linked sugars based on resistance to endoglycosidase F, d) resistance to trypsin and chymotrypsin digestion but susceptibility to pronase, and e) presence of sialic acid and lactosaminoglycan as O-linked sugars. Gp140 could be labeled with the periodate/NaB[3H]4 technique, indicating its similarity to a class of sialoglycoproteins previously described on activated T-cells in mouse and man. The antigenic epitope recognized by MAb T305 contains sialic acid linked (2----3) to galactose; however, periodate oxidation of the exocyclic ring of sialic acid did not affect binding by MAb T305. In an attempt to determine the functional role of gp140, we tested the ability of MAb T305 to block: a) proliferation of peripheral blood lymphocytes to mitogens, b) response to interleukin 2 (IL 2) of an IL 2 dependent T cell line, and c) growth of a T-ALL derived cell line. No inhibition of proliferation or growth was noted. Although the function of gp140 remains unknown, its association with lymphocyte activation and certain disease states suggests that it may provide a target for modulation of the immune response. These studies characterize the structural features of gp140 and further define the epitope recognized by MAb T305. PMID- 2987345 TI - Immunoactive products of placenta. IV. Impairment by placental cells and their products of CTL function at effector stage. AB - Trophoblast-enriched cell suspensions prepared by collagenase digestion from midterm murine placentae were found resistant to CTL-mediated lysis. Treatment of such cells by trypsin or neuraminidase rendered these cells susceptible to such lytic effectors. Collagenase-prepared cell suspensions could impair CTL action, whereas neuraminidase- or trypsin-treated cells did not retain this property. This effect was also observed with extracts. These results indicate that soluble factors (which we will characterize in another paper) released by trophoblast cells (in fact, spongiotrophoblast) can interfere in a dose-dependent fashion with the action of lytic effectors. We suggest that such active mechanisms are physiologic components of the placental barrier and might be defective in some cases of immunologic abortions. PMID- 2987346 TI - Monoclonal antibodies reactive with human T cell lymphotropic virusI (HTLVI) p19 internal core protein: cross-reactivity with normal tissues and differential reactivity with HTLV types I and II. AB - Three monoclonal antibodies to human T cell lymphotropic virus type I (HTLVI) p19 internal core protein, designated as alpha HTLV-2, 3, and 4, have been developed. In indirect immunofluorescence (IF) assays, these antibodies reacted with acetone fixed cytocentrifuge preparations of culture HTLVI-infected peripheral blood leukocyte (PBL) from a patient (SD) with Japanese adult T cell leukemia and with infected HUT-102 T cells but not with cultured normal PBL. Anti-p19 antibodies alpha HTLV-2, 3, and 4 all reacted with the same HTLVI p19 identified both by antibodies in HTLVI+ patient sera and by antisera raised against two synthetic peptides encoded by the p19 gag region of HTLVI. Partial proteolytic cleavage of p19 immunoprecipitates obtained with antibodies alpha HTLV-2, 3, and 4 produced a 17,000-dalton cleavage product, in agreement with the size of the fragment predicted from the nucleic acid sequence of the HTLVI p19 gag region. Antibodies alpha HTLV-3 and 4 reacted with HTLVI but not HTLVII proteins and were useful diagnostic probes in identifying HTLVI- but not HTLVII-infected lymphoid cells in immunofluorescence assays. In addition to reacting with HTLVI p19, antibodies 2 and 4 also cross-reacted with a wide variety of HTLV-uninfected normal and neoplastic cells and tissues. In addition, HTLVI+ patient sera contained antibodies that competed for binding to the antigenic site on p19 recognized by antibody 4. Thus, anti-p19 monoclonal antibodies alpha HTLV-2 and 4 reacted with a 19,000-dalton viral-encoded protein of HTLVI and cross-reacted with normal host tissues, while anti-p19 antibody alpha HTLV-3 was specific for HTLVI p19 core protein. PMID- 2987347 TI - IL 2 receptor induction on human T lymphocytes: role for IL 2 and monocytes. AB - In this report we studied the requirements for the activation and proliferation of highly purified human T lymphocytes. Purified T cells incubated for 3 days with PHA neither proliferate nor express IL 2 receptors as detected by FACS analysis with the use of anti-Tac antibodies. However, purified T cells incubated with Con A or anti-T3 moAb do not proliferate, albeit 30 to 35% T cells express Tac epitopes. The addition of IL 2, either natural purified or recombinant, resulted in both the appearance of Tac antigen and the proliferation of PHA activated T cells. Much to our surprise, IL 2 did not induce proliferation of Tac positive T cells activated by Con A or soluble anti-T3 unless monocytes were added to the cultures. These data suggested that two classes of IL 2 receptors might exist on T cells, one of which was not functionally involved in T cell proliferation. In keeping with this interpretation, we have been able to demonstrate, using a radiolabeled IL 2 binding assay, that anti-T3 moAb induced almost exclusively IL 2 receptors of low affinity (Kd = 30 to 70 X 10(-9) M) and that additional signals, provided by monocytes, are required for the acquisition of high affinity receptors. IL 2 itself can induce high affinity receptors on PHA stimulated T cells but not on cells activated by Con A or anti-T3. In this latter case the physical presence of monocytes is required and cannot be substituted by IL 1, thus indicating a previously unreported role for monocytes. It is postulated that the contact of monocytes with T cells induces a switch from an inactive low affinity conformation of the IL 2 receptor to a functional high affinity one. PMID- 2987348 TI - Studies of cytochrome b-245 translocation in the PMA stimulation of the human neutrophil NADPH-oxidase. AB - The human neutrophil respiratory burst, activated by phorbol 12-myristate 13 acetate (PMA), results from specific receptor-ligand binding and activation of the NADPH-oxidase in the plasma membrane. The role of granule membrane constituents has been elucidated with neutrophils disrupted by nitrogen cavitation and then fractionated in Percoll gradients to resolve four postnuclear fractions: cytoplasm, light membranes or gamma fraction (site of the NADPH oxidase), a light granule (beta) fraction containing putative constituents of the NADPH-oxidase (cytochrome b-245 and an associated flavoprotein), and a fraction of heavy granules. Cytochrome b-245 is localized to two pools of specific granules within the beta fraction as assessed by differing sedimentation in narrow Percoll gradients and translocates upon PMA-stimulation from one of these specific granule sub-pools to the plasma membrane where it exhibits no change in its midpoint redox potential. Translocation of cytochrome b-245 parallels O2 production initiated by PMA stimulation as assessed in the time course of each activity. The finding of increased amounts of the b cytochrome in cytoplast membranes relative to plasma membranes of unstimulated cells suggests that the cytoplasts, devoid of granules yet capable of O2-generation upon PMA-stimulation, are useful in assessing post-translocation events in the activation pathway of the NADPH-oxidase. These data support the hypothesis that translocation of NADPH oxidase components from an intracellular granular pool contributes to respiratory burst expression. PMID- 2987349 TI - Transduction by leukotriene B4 receptors of increases in cytosolic calcium in human polymorphonuclear leukocytes. AB - The uptake of Quin-2 by human polymorphonuclear (PMN) leukocytes permitted accurate fluorimetric quantification of the cytosolic concentration of intracellular calcium [( Ca+2]in), without altering the expression of the two subsets of leukotriene B4 (LTB4) receptors, as assessed by the binding of [3H]LTB4. Chemotactic concentrations of LTB4 elicited a rapid increase in [Ca+2]in, which reached a peak within 0.6 to 1 min and then decayed back to baseline levels by 6 to 10 min. The maximal increase and the half-maximal increase in [Ca+2]in were achieved by LTB4 at mean concentrations of 5 X 10(-10) M and 2 X 10(-10) M, respectively, where the binding of LTB4 to high-affinity receptors predominates. A rank order of potency of LTB4 greater than 5(S),12(S)-6 trans-LTB4 greater than 12(S)-LTB4 was established for the elicitation of increases in [Ca+2]in, which reflects the binding of the isomers to low-affinity receptors. PMN leukocytes were preincubated with 10(-8) M LTB4 to induce chemotactic deactivation, which eliminates the expression of high-affinity receptors without altering the expression of the low-affinity receptors for LTB4. LTB4 elicited an increase in [Ca+2]in in the deactivated PMN leukocytes with an EC50 of 3 X 10(-8) M, which is similar to the Kd for LTB4 binding to the low affinity receptors. Two lines of cultured human leukemic cells, IM-9 and HL-60, did not bind LTB4 specifically and did not show any change in [Ca+2]in upon the addition of 3 X 10(-8) M LTB4. The HL-60 human promyelocytic leukemia cell line was induced to differentiate in 1% dimethyl sulfoxide to leukocytes with more mature myelocytic characteristics. Differentiated HL-60 cells expressed an average of 54,000 low-affinity receptors for LTB4 per cell with an average dissociation constant of 7.3 X 10(-8) M and concurrently developed the capacity to respond to LTB4 with an increase in [Ca+2]in. The binding of LTB4 to either high-affinity or low-affinity receptors appears to be sufficient to initiate an increase in [Ca+2]in in human PMN leukocytes and differentiated HL-60 cells. The specificity of LTB4 receptors in transducing maximum increases in [Ca+2]in is determined by the subset of receptors that predominate as a result of the concentration of LTB4 and the state of the responding cells. PMID- 2987350 TI - Retrovirus-mediated immunosuppression. II. FeLV-UV alters in vitro murine T lymphocyte behavior by reversibly impairing lymphokine secretion. AB - Murine splenocytes and cloned murine T cells were used to study the in vitro immunosuppressive effects of UV-inactivated feline leukemia virus (FeLV-UV) on lymphokine secretion. FeLV-UV can significantly depress the accumulation of IL 2 in cultures of Con A-stimulated C57BL/6 splenocytes and in cultures containing the alloreactive helper T cell clone B6D/2-2m plus Con A. Inhibition of lymphokine accumulation in these cultures could not be attributed to absorption or inactivation of IL 2 by the FeLV-UV or to the FeLV-UV-induced production of substances which interfere with the IL 2 bioassay. Thus, FeLV-UV appears to block production and/or secretion of IL 2 by a direct inhibitory effect on IL 2 secreting murine T lymphocytes. Additional studies indicate that FeLV-UV impairs IL 2 production only if added very soon after lymphocyte contact with lymphokine inducing agents and that IL 2 secretion resumes when FeLV-UV is removed from the culture. FeLV-UV also impairs accumulation of MAF (interferon-gamma?) in cultures of Con A-stimulated C57BL/6 splenocytes and in cultures containing the alloreactive cytotoxic T lymphocyte clone B6D/2-7c plus Con A. The latter observation again suggests that FeLV-UV impairs lymphokine secretion by a direct effect on lymphokine-producing T lymphocytes. Furthermore, it suggests that FeLV UV does not selectively impair production of IL 2 nor does it have selective inhibitory effects on helper T cells. Rather, FeLV-UV appears to have a general inhibitory effect on lymphokine production by T lymphocytes. Finally, concentrations of FeLV-UV which suppress MAF production by the CTL clone have little influence on the cytolysis mediated by the same cloned T cell population. Thus, the immunosuppressive influence of FeLV-UV is selective for phenomena associated with induction of new T lymphocyte functions, such as lymphokine secretion, and spares other immune functions already expressed by the same cells. PMID- 2987351 TI - Induction of protective immune response in cats by vaccination with feline leukemia virus iscom. AB - An effective candidate subunit vaccine consisting of the gp 70/85 of feline leukemia virus (FeLV) was prepared by using the immunostimulating complex (iscom) method for the presentation of membrane proteins of enveloped viruses. Two 32-wk old specific pathogen-free (SPF) cats were immunized with a FeLV iscom vaccine prepared from the supernatant fluid of the FL74 tumor cell line without adjuvant. Both cats developed FeLV serum antibodies, as measured in an enzyme-linked immunosorbent assay (ELISA) and in a virus neutralization test. A proportion of the antibodies were directed to an epitope located on gp70/85, which was shown in competition ELISA with a peroxidase-labeled virus-neutralizing monoclonal antibody to be shared by all three subtypes of FeLV. The protective effect of FeLV iscom was studied by vaccinating six 8-wk-old SPF cats with iscom prepared from cell culture supernatant of another tumor cell line F422, followed by oronasal challenge with 10(6) ffu FeLV-A (strain Glasgow-1). Six unvaccinated cats were also challenged with the same dose of FeLV. The vaccinated cats developed FeLV serum antibodies, some of which were directed to the shared epitope on gp70/85. At 10 wk after challenge, none was viremic, whereas three of the control cats had developed FeLV viremia. The potential of FeLV iscom as a vaccine against FeLV-associated disease in cats, and of iscom vaccines for protection against mammalian retrovirus infections, is discussed. PMID- 2987352 TI - Mouse immunoglobulin gene rearrangements. The sequence of a nonexpressed lambda 3 chain gene. AB - A functionally defective lambda 3-immunoglobulin chain gene has been cloned from plasmacytoma HOPC-1 (gamma 2b, lambda 1). The lambda 3 gene resulted from the juxtaposition of the germline V lambda 1 sequence with a J lambda 3 C lambda 3 gene segment. DNA sequencing of the rearranged V lambda 1 J lambda 3 exon showed the presence of a single base pair deletion at the site of V-J joining. The alteration in the reading frame caused by this deletion generated a stop codon at the 3' end of J lambda 3, thus rendering this gene nonfunctional for light chain production. In addition, a one-point mutation in the J lambda 3-C lambda 3 intron distinguishes the rearranged gene from the unrearranged counterpart. The implications that this rearrangement has in terms of the mechanism of somatic mutations and of selective proliferation of B cells mediated by antigen stimulation are discussed. PMID- 2987353 TI - Complete nucleotide sequence of the murine gamma 3 switch region and analysis of switch recombination sites in two gamma 3-expressing hybridomas. AB - The heavy chain isotype switch is mediated by a DNA rearrangement between a donor switch region (usually mu) and a recipient switch region (gamma, epsilon, or alpha). Switch regions lie upstream of the appropriate heavy chain constant region gene and are composed of simple sequences repeated in tandem. It is not known to what extent the tandemly repeated sequences are important to the heavy chain switch recombination, and to what extent other features of switch region sequences might contribute to the switch process. We studied switches to the gamma 3 isotype by sequencing the entire gamma 3 switch region. This switch region is composed of forty-four 49 base pair units repeated in tandem. These repeated units share modest homology with the mu switch region repeated elements. Evolution of the gamma 3 switch region seems to involve insertions and deletions of the 49mer elements. We also molecularly cloned rearranged switch regions from two gamma 3-expressing hybridomas and determined the DNA sequences at the mu gamma 3 recombination sites. We located these switch recombination sites within the germ-line gamma 3 switch region, as well as switch recombination sites from two myelomas. All four sites are found in the 5' one-third of the gamma 3 switch region. We discuss some additional trends in the sequence data near these four recombination sites. PMID- 2987354 TI - A monoclonal antibody that recognizes an Ly-6-linked antigen inhibits the generation of functionally active T cell subsets. AB - A monoclonal antibody (mAb) generated against the chemically-induced BALB/c Meth A sarcoma, designated HD42, reacts in cytotoxic tests with Meth A as well as with BALB/c peripheral lymph node cells and mitogen-activated spleen cells. The antigen was detected by FACS analysis on BALB/c spleen and lymph node cells, and by absorption assays on all normal lymphoid cells of BALB/c but not B6 mice. The expression of the antigen was not found on normal adult lung fibroblasts, on brain, nor on an extensive panel of tumors of BALB/c and B6 origin. Because the strain distribution of the antigen is reciprocal to that of Ly-6.2 and is not expressed in congenic C3H.Ly-6b mice, we have tentatively defined it as Ly-6.1 and referred to the mAb as alpha-Ly-6.1. The presence of alpha-Ly-6.1 abrogates both the Con A-induced and the IL 2-dependent proliferative response of normal T cells, whereas the response of normal B cells to LPS remains unaffected. alpha-Ly 6.1 is a potent suppressor of the primary in vitro plaque-forming cell (PFC) response to SRBC. Pretreatment of normal splenic T cells with alpha-Ly-6.1 and complement had no effect on the ability of these cells to generate in vitro either T helper cells (TH) or T suppressor cells (TS) to SRBC. However, addition of antibody in the absence of complement during the generation of TH or TS, or posttreatment of these T cell subsets with antibody and complement after in vitro education, completely removed the functional activity of these cell types. Addition of alpha-Ly-6.1 to MLC suppressed the MLR as well as the generation of cytotoxic lymphocytes (CTL), whereas the presence of the antibody during a cell mediated lympholysis (CML) had no effect. Therefore, it appears that alpha-Ly-6.1 recognizes an antigen that is important for the generation of TH and TS cell subsets. PMID- 2987355 TI - HLA-DR4-associated haplotypes are genotypically diverse within HLA. AB - Biochemical diversity among products of class II HLA genes has been observed in individuals who appear to be HLA-D and DR-identical by cellular and serologic typing. We used techniques of restriction enzyme fragment analysis by Southern blotting to analyze this diversity at the level of cellular DNA. A panel of 17 HLA-DR4 homozygous cell lines (HCL) were investigated by using cDNA probes homologous to DQ beta, DQ alpha, and DR beta genes. Each probe was hybridized to cellular DNA digested with a series of different restriction endonucleases. Polymorphisms were observed with the use of the enzymes Pst I, Hind III, and Bam HI: Hybridization of cellular DNA digested with Hind III and Pst I with the DQ beta probe revealed specific polymorphisms, as did hybridization of the Pst I digest with the DQ alpha cDNA probe and the Bam HI digest with the DR beta probe. The observed differences fall into two categories: first, considerable diversity was seen between HLA-DR4 HCL that represent different HLA-D-defined haplotypes; second, diversity was also observed among HCL of the same DR4-associated HLA-D cluster. In contrast to the DQ cDNA probes, hybridization with the DR beta probe revealed relatively limited polymorphism by using a panel of different restriction endonucleases. Thus, although there is a general pattern of polymorphic restriction enzyme fragments homologous to DQ probes within an HLA-D cluster, the pattern seen for any particular cell line was not sufficiently distinct to assign an HLA-D or DR specificity. PMID- 2987356 TI - Immune induction of human monocyte plasminogen activator. Characteristics of an assay for cell-mediated immunity. AB - We characterized immunologic induction of monocyte plasminogen activator (PA) to determine whether assay for PA induction reliably detected cell-mediated immunity (CMI). Mononuclear leukocytes (MNL) were incubated in teflon-lined culture tubes for 1-4 days in the presence or absence of phytohemagglutinin-P (PHA), concanavalin A (Con A) or Candida antigen. PA activity of the monocytes in those suspensions was then measured using a micro fibrin plate assay. Monocytes in stimulated MNL had more PA activity than monocytes in unstimulated MNL. Maximal differences between stimulated and unstimulated cells were seen after 2 days of culture. Dose-response studies demonstrated that PA induction occurred at submitogenic concentrations of stimuli. Peak induction was seen using suboptimally mitogenic concentrations of PHA, Con A and Candida antigen. PA induction in response to Candida stimulation corresponded with skin test results. More than 90% of healthy adults tested had positive assays to all stimuli. LPS, in picogram concentrations, induced PA activity in the absence of lymphocytes, but such induction was prevented by polymyxin B. Supernates from activated MNL also induced PA in purified monocytes. This indirect assay of PA induction was less sensitive than direct assay of the MNL. A standard indirect assay for leukocyte inhibitory factor (LIF) was also less sensitive than the direct PA induction assay. The direct PA induction assay is sensitive and convenient and requires small volumes of blood. It may prove valuable in in vitro analysis of cell-mediated immunity in health and disease. PMID- 2987358 TI - A rapid and efficient microtechnique for the analysis of functional transferrin receptors on tumor cells. AB - A simple and efficient method for the analysis of the affinity and number of functional transferrin receptors (TFR) on human tumor cells is described. The technique is designed to utilize microtitration equipment; and is suitable for easy comparison of up to 8 different cell preparations per assay. Using this technique, 5 established cell lines were evaluated for functional TFR expression. The control erythroleukemic cell line K562 possessed 3.28 X 10(5) functional TFR per cell (+/- 3.69 X 10(4), SEM) Kd = 9.0 X 10(-9) X M-1. Trypsin and heat pretreated cells were compared to control erythroleukemic K562 cells from the same culture to determine both the effects of receptor removal and cell viability on the assay. Trypsin and heat pretreatment of these K562 cells severely decreased receptor function as indicated by Scatchard analysis as well as by time course and cold competition analysis respectively. Whereas the affinity of trypsin-treated receptors on cells was similar to control values, heat-killed cells displayed an altered cellular affinity for 125I-transferrin underscoring the importance of utilizing cells of high viability in receptor assays. PMID- 2987357 TI - A simple panning method for the selection of cell surface antigen transfectants. AB - Mouse Ltk- (H-2k) cells that became reactive with anti-H-2d monoclonal antibodies (McAb) after transfection with DNA from H-2d-positive cells were isolated by an indirect panning method. The cells were transfected with DNA from a plasmid containing the selective thymidine kinase (tk) gene in addition to total cell DNA, so that a first selection could be carried out in hypoxanthine/aminopterin/thymidine (HAT) medium. The HAT-selected tk+ transfectants were incubated with anti-H-2d McAb, washed and transferred to dishes coated with purified anti-immunoglobulin (anti-Ig). This 2-step method of panning has the advantage that only the anti-Ig reagent requires purification. Transfected cells clearly reactive with either anti-Ld/Rd or anti-Kd McAb were isolated after only 1 or 2 cycles of panning. PMID- 2987359 TI - A whole-blood lymphoproliferation assay for measuring cellular immunity against herpes viruses. AB - A whole blood test system was established to study cell-mediated immunity to cytomegalovirus (CMV) and herpes simplex virus (HSV) in a large number of healthy blood donors. Cellular immunity was measured by the in vitro proliferative response (LP) of peripheral lymphocytes. These responded vigorously to several mitogens. Lymphocytes of most individuals responded to HSV, but only a limited number were reactive towards CMV. In parallel, antibodies against CMV and HSV were measured by an ELISA technique. For HSV, good correlation was observed between serological and lymphocyte proliferation results. For CMV, no clear correlation was obtained, only 21 of 40 donors positive in the antibody test being positive in the LP test. The majority of seronegatives were negative in the LP test. Use of virions purified by sucrose gradient centrifugation, or an additional strain of CMV (strain Davis) did not increase the number of donors positive in the LP test. One explanation might be that individuals possessing antibodies against CMV as measured by ELISA but no capacity to react in the LP test had suffered from a CMV infection a long time before, and now showed waning cellular immunity, but antibody still detectable. Use of the whole blood technique on 108 individuals showed that this very simple test works well with various mitogens and at least some antigens. PMID- 2987360 TI - Ultraviolet radiation augments epidermal beta-adrenergic adenylate cyclase response. AB - Pig skin was irradiated in vivo with fluorescent sunlamp tubes (peak emission at 305 nm). A significant increase in epidermal beta-adrenergic adenylate cyclase response was observed as early as 12 h following 1-2 minimum erythema doses (MEDs) UVB exposure, which lasted at least 48 h. The augmentation of adenylate cyclase response was relatively specific to the beta-adrenergic system and there was no significant difference in either adenosine- or histamine-adenylate cyclase response of epidermis. The increased beta-adrenergic adenylate cyclase response was less marked at higher doses of UVB exposure (5 MEDs); in the latter condition, a significant reduction in adenosine- or histamine-adenylate cyclase response was observed. There was no significant difference in either low- or high Km cyclic AMP phosphodiesterase activity between control and UVB-treated skin at 1-2 MEDs. Our data indicate that the epidermal adenylate cyclase responses are affected in vivo by UVB irradiation, which might be a significant regulatory mechanism of epidermal cyclic AMP systems. PMID- 2987361 TI - Analysis of oral papillomas, leukoplakias, and invasive carcinomas for human papillomavirus type related DNA. AB - Five papillomas, five leukoplakias, and six carcinomas were investigated for the presence of papillomavirus group-specific antigens and viral DNA. Viral proteins were identified with genus-specific papillomavirus antibodies. Cloned human papillomavirus (HPV) 11 and 16 DNA were used as probes in Southern blot hybridization at conditions of different stringency in order to determine viral DNA. Four of five papillomas, four of five leukoplakias, and three of six carcinomas reacted with HPV DNA probes and revealed some stained cells after exposure to HPV antibodies. HPV type 16 was found in one carcinoma and HPV type 11 was demonstrated in another case of carcinoma. PMID- 2987363 TI - Possible functions of 1 alpha,25-dihydroxyvitamin D3, an active form of vitamin D3, in the differentiation and development of skin. PMID- 2987362 TI - The responses of normal and psoriatic skin to single and multiple topical applications of leukotriene B4. AB - Topical application of single doses of leukotriene B4, (5-500 ng) under occlusion to the skin of normal volunteers and to the clinically normal skin of patients with untreated stable plaque psoriasis caused an inflammatory reaction lasting several days. Histologically, intraepidermal neutrophil microabscesses were seen in both groups following 50 ng applications of leukotriene B4, but there were significantly fewer intraepidermal neutrophils in the psoriatic than in the normal subjects. Nine repeated daily applications of leukotriene B4 (50 ng) to the same site were associated with a reduction of the visible inflammatory response in both volunteer groups, although application of a single 50 ng dose of leukotriene B4 to an adjacent, previously untreated site at the end of the multiple dose experiments elicited acute erythema and edema. Histologically, there were significantly fewer intraepidermal neutrophils in biopsies from sites to which 9 applications of leukotriene B4 were made than in sites to which 1 dose had been applied at the end of the multiple-dose experiments in both psoriatic and normal volunteers. Therefore, repeated application of leukotriene B4 is associated with decreased responsiveness which is due to a local mechanism in the repeatedly treated skin. No subject developed psoriasis following single or multiple applications of leukotriene B4, suggesting that the production of leukotriene B4 in the epidermis is unlikely to be a primary pathogenic event in this disease. PMID- 2987364 TI - Detection and developmental changes of the 1,25-(OH)2-D3 receptor concentration in mouse skin and intestine. AB - We investigated [3H]1,25-dihydroxyvitamin D3-specific binding activity in fetal, neonatal, and adult mouse skin to determine (a) during which stage in development the skin develops the capacity to respond to this hormone and (b) whether the hormone binding activity changed during development and maturation. A macromolecule with properties similar to those of the chick intestinal 1,25 dihydroxyvitamin D3 receptor was detected in the skin and intestine of mouse pups at 17 days of fetal life. 1,25-Dihydroxyvitamin D3-specific binding activity from both tissues sedimented on linear sucrose gradients at 3.5-3.7S and eluted from DNA cellulose at 0.22 M KCl. At earlier stages of fetal life (12-14 days) receptor-like activity was detected in cytosols prepared from whole-mouse fetuses. 1,25-Dihydroxyvitamin D3-specific binding activity was quantitated in the skin and intestine throughout development using a chromatin binding assay. Scatchard analysis of saturation binding data showed that the concentration of binding activity in skin increased rapidly after birth and reached a maximum when the mice were 10-19 days old. By contrast, the binding activity that was detected in the fetal and neonatal whole intestine remained low until the mice were weaned. The affinity (Kd) of the binding activity was similar in skin and intestine at all ages studied. It is concluded that 1,25-dihydroxyvitamin D3 specific binding activity appears in both skin and intestine of the mouse prior to birth and increases in these two tissues during different stages in development. PMID- 2987366 TI - Hepatitis B viral DNA in infected human liver and in hepatocellular carcinoma. AB - Blot-hybridization analysis of DNA forms from hepatitis B virus (HBV) in the liver of chronic carriers of hepatitis B surface antigen (HBsAg) has revealed the presence of both relaxed and closed circular viral DNA and of novel viral DNA-RNA hybrid molecules in patients with complete virions, with the hepatitis B e antigen (HBeAg), or with both, in serum. One carrier of HBsAg with no detectable virus or HBeAg in his serum had small amounts of free viral DNA in his liver sample, a finding suggesting the potential for production of complete virus; another such carrier had only HBV DNA integrated in cellular DNA and, thus, may have lost the ability to replicate virus. The liver sample of one of eight patients with antibodies to HBsAg in his serum, but no HBsAg, contained small amounts of free viral DNA. Analysis of tissue from hepatocellular carcinoma revealed evidence for integrated viral DNA sequences in multiple-cellular DNA sites, and stoichiometric analysis suggested that the tumors were monoclonal in origin. These results demonstrate the presence of a new form of HBV in infected human liver and reveal that serological profiles are not always a reliable guide in determining the presence of potentially infectious forms of HBV in the liver. PMID- 2987365 TI - Molecular epidemiology of plasmid-mediated ampicillin resistance in Haemophilus influenzae type b isolates from Alaska. AB - Native Alaskans have a high incidence of disease caused by invasive Haemophilus influenzae type b and represent an isolated population for epidemiological study. We used plasmid DNA analysis and subtyping of outer membrane proteins as markers to characterize 29 ampicillin-resistant, invasive strains and seven ampicillin resistant, noninvasive strains of this organism from distinct geographic regions. All 36 strains produced beta-lactamase; 34 strains transferred resistance by conjugation. Seven of the 36 strains harbored detectable plasmid DNA: four had a molecular mass of 40 MDa, and three had a molecular mass of 3 MDa. Furthermore, 20 transconjugants had a similar large plasmid, and four had a similar small plasmid. Ten of 12 transconjugants with either the large, small, or undetectable plasmid DNA were able to retransfer resistance. Transformation of resistance was successful with two large plasmids. DNA-DNA hybridization studies revealed that 33 of 36 strains had DNA homology. Restriction endonuclease digestion of 10 large plasmids revealed five patterns; identity was evident within a geographic region, and similarity existed between regions. Seven restricted plasmids demonstrated an identical pattern with a small beta-lactamase probe. Ampicillin resistance in these isolates from Alaska is primarily due to a common, 40-MDa conjugative plasmid that mediates beta-lactamase production, a finding which differs from that for ampicillin-resistant plasmids isolated elsewhere in the United States. Despite variable outer membrane protein profiles of the distinct strains of H. influenzae type b, the plasmids shared significant DNA homology. It appears that a common genetic element was responsible for the dissemination of this phenotype for resistance in Alaska. PMID- 2987367 TI - Kinetics of IgM and IgG responses to Japanese encephalitis virus in human serum and cerebrospinal fluid. AB - We measured levels of antibodies to Japanese encephalitis virus (JEV) in serum and in cerebrospinal fluid (CSF) specimens obtained from 32 patients with acute encephalitis by using "antibody-capture" solid-phase enzyme-linked immunoassays specific for IgM or IgG to JEV. The proportions of confirmed cases with IgM to JEV detectable in CSF were 68% (obtained on day 1), 100% (day 7), 96% (day 30), and 72% (day 180). For IgG in CSF the proportions were 47% (day 1), 89% (day 7), 100% (day 30), and 100% (day 180). Twenty-five CSF samples were obtained from control patients with other diseases with possible nervous system involvement (but none with a clinical diagnosis of viral encephalitis); none had detectable IgM to JEV. Five JEV-infected but asymptomatic siblings of patients with encephalitis were also examined; all had high levels of IgM to JEV in serum, but none had detectable IgM to JEV in CSF. PMID- 2987368 TI - Systemic lymphoid atrophy in coxsackievirus B3-infected mice: effects of virus and immunopotentiating agents. AB - Adult mice infected with coxsackievirus B3 (CB3) showed a generalized lymphoid involution. This effect was produced by most of the isolates and clones of CB3 and, to a lesser extent, by coxsackievirus B1 but not by other highly pathogenic picornaviruses. While moderate involution of the thymus also occurred in mild CB3 infections, peripheral changes strictly correlated with the severity of disease. The major alteration of the thymus was a massive cortical depletion, whereas the reduction of spleen and lymph node cellularity appeared devoid of selectivity. No histological and ultrastructural signs of CB3 replication in such organs could be detected. Attempts to demonstrate CB3 replication in thymus, spleen, and lymph node cells were unsuccessful. CB3-induced lymphoid atrophy was not prevented by adrenalectomy and was exacerbated by three different immunopotentiating agents. Thus, certain picornaviruses may cause severe stress-independent lymphoid damage in the absence of viral replication in situ. A role for autoreactive mechanisms is suggested. PMID- 2987369 TI - Synergism between recombinant human interferon and nucleoside antiviral agents against herpes simplex virus: examination with an automated microtiter plate assay. AB - An automated, quantitative, cytopathic effect (CPE) inhibition assay with human fibroblasts in 96-well microtiter plates was used to examine the combination of recombinant human interferon-alpha (rIFN-alpha A) and acyclovir, vidarabine, or dihydroxypropoxymethyl guanine against herpes simplex virus types 1 (HSV-1) and 2 (HSV-2) in vitro. Fifty percent CPE (CPE50) end points, calculated from optical density readings of crystal violet-stained monolayers in an automated spectrophotometer, represented 1.7 log reduction in viral yield (50-fold or 98% decrease). Using CPE50 end points of drugs alone and in combination, we defined synergism, additivism, or antagonism with an isobologram plot and a combination index equation. The combinations of rIFN-alpha A plus acyclovir and rIFN-alpha A plus dihydroxypropoxymethyl guanine were highly synergistic against both HSV-1 and HSV-2, whereas the combination of rIFN-alpha A plus vidarabine was additive to mildly synergistic. Combinations of antiviral agents synergistic in cell cultures should be pursued with further studies in animal models of human viral disease and potentially in clinical trials. PMID- 2987370 TI - Long-term serological follow-up of patients for Epstein-Barr virus after recovery from infectious mononucleosis. PMID- 2987371 TI - Serological testing for Epstein-Barr virus--developments in interpretation. PMID- 2987372 TI - [Effect of cyclophosphamide treatment on mice infected with Japanese encephalitis virus]. PMID- 2987373 TI - [Primary mediastinal germ cell tumor: a report of four cases]. PMID- 2987374 TI - [Receptor activation and inositol phospholipids]. PMID- 2987375 TI - Studies on the placental transport mechanism for iron. AB - Transferrin (Tf) and ferritin (Fr), the proteins which participate in iron transport, were examined to determine their fluctuation during pregnancy and their localization in human placental tissue, and the iron transport mechanism between mother and fetus was discussed. The main results are as follows: Maternal serum ferritin (SFr) decreased remarkably and the maternal total iron binding capacity (TIBC) increased gradually as pregnancy progressed. Maternal serum iron (SI), SFr and TIBC at delivery were 59.8 +/- 26.2 micrograms/dl, 9.6 +/- 7.2 ng/ml and 495.2 +/- 100.3 micrograms/dl, and cord blood SI, SFr and TIBC were 161.5 +/- 42.1 micrograms/dl, 160.5 +/- 67.2 ng/ml and 177.7 +/- 33.9 micrograms/dl, respectively. Peroxidase conjugating antibody method revealed the localization of Tf on the microvillous surface of syncytiotrophoblasts, and the localization of Fr in all layers of trophoblasts, especially in the neighborhood of the surface. Ferritin content of the placenta in the 1st, 2nd and 3rd trimesters was 0.07 +/- 0.03, 0.18 +/- 0.03 and 0.25 +/- 0.09 (micrograms/mg protein), respectively. These results indicate that iron is transferred from the mother to the fetus by the placental active function, and Tf receptor and Fr on trophoblasts participate in the adequate placental iron transport. PMID- 2987376 TI - The influence of immunosuppression and immunodeficiency on infections with leprosy and tuberculosis. PMID- 2987377 TI - [An epidemiological study on dietary ingestion of eicosapentaenoic acid (EPA) and platelet function in Japanese]. PMID- 2987378 TI - Free vascularised transfer of a metatarsophalangeal joint to the hand. A technical modification. AB - We describe the vascularised transfer of a second metatarsophalangeal joint to the hand to correct a congenital deformity in a thirteen-year-old boy. Technical refinements are described, allowing inversion of the joint to the optimal position for subsequent mobility and retention of a large overlying dorsalis pedis flap to provide adequate cover. PMID- 2987379 TI - Collagen synthesis and collagenase activity in dermal fibroblasts from patients with diabetes and digital sclerosis. AB - Collagen production and collagenase activity were measured in dermal fibroblast cultures obtained from eight patients with diabetes with digital sclerosis and three normal controls. Total collagen synthesis in patients with diabetes was reduced in comparison with controls. DNA replication was also reduced in patients with diabetes. No differences in collagenase activity were noted. Our results suggest that, in contrast to systemic sclerosis, increased synthesis does not contribute to the collagen accumulation of diabetic digital sclerosis. Decreased degradation related to nonenzymatic glucosylation of collagen is a more likely mechanism. PMID- 2987380 TI - Enzyme immunoassay of anti-glomerular basement membrane antibodies. AB - An immunoassay has been developed to detect anti-glomerular basement membrane (GBM) antibodies in human sera. Various plating conditions, types of microtiter plates, and the use of biotinylated or peroxidase-labeled secondary antibodies were examined. The described assay is reliable, fast, and convenient. Sera with positive reactivity in anti-GBM nephritis and Goodpasture's syndrome are readily discriminated from sera obtained from normal individuals or patients with a variety of other diseases. PMID- 2987381 TI - Radioreceptor assay for beta-adrenergic receptor-reactive catecholamines in serum samples. AB - A procedure for effective application of ultrafiltration is described for preparation of serum catecholamine samples suitable for a sensitive radioreceptor assay. Each of the fresh serum samples unsuitable for the direct receptor assay was dialyzed by applying centrifugal force into a capsule-shaped porous core whose surface was tightly covered with a molecular filtration membrane. Interfering macromolecular serum components were left outside the capsule during the centrifugation or forced dialysis. The ultrafiltrate in the capsule was readily recovered by another centrifugation, and was found satisfactory as an assay specimen. In the radioreceptor assay system adapted to detect 30 pg per assay tube, epinephrine added to fresh serum to make the final concentration approximately 1 X 10(-8) mol/L was recovered semiquantitatively. Serum catecholamine level determined was equivalent to beta-adrenergic receptor reactivity that could be expressed in terms of (-)-epinephrine concentration. PMID- 2987382 TI - Inhibition of plasminogen activators by conditioned medium of human hepatocytes and hepatoma cell line Hep G2. AB - Primary cultures of human hepatocytes and the human hepatocellular cell line Hep G2 are shown to produce fast-acting inhibitors of tissue-type plasminogen activator (tPA) and urokinase. The tPA inhibitory activities in conditioned medium of these liver cell types are very similar to those present in human endothelial cell conditioned medium. They are stable at pH 2.5, have similar dissociation constants with tPA (1.5 to 5 pmol/L), and are similar in thermostability. Addition of tPA to conditioned medium of Hep G2 and endothelial cells that has been depleted of tPA and urokinase reveals a 100 kilodalton tPA inhibitor complex. The fast-acting tPA inhibitory activity in human plasma has comparable properties, and may originate from the liver or the vascular endothelium or both. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis of conditioned medium from hepatocytes, Hep G2, and endothelial cells, additional fibrinolytic inhibition at 52 kilodaltons was visualized. This was not found with human plasma. PMID- 2987384 TI - Malignant fibrous histiocytoma of maxilla following radiotherapy for bilateral retinoblastoma. AB - A 20-year-old man developed a malignant fibrous histiocytoma in the right maxilla 19 years after irradiation for bilateral retinoblastoma. The incidence of second tumours in patients who survived bilateral retinoblastoma treated with radiation was 8.5 per cent. Malignant fibrous histiocytomas which arise in a site of prior radiation are fatal. The present case is presumed to have the autosomal dominant retinoblastoma gene, not associated with deletion of the q 14 band of chromosome 13. The patient succumbed to the second tumour. PMID- 2987383 TI - Paraganglioma of the tympanic membrane. AB - Von Haller, in 1743, first described the carotid body and was so impressed with the richness of its innervation that he called it 'ganglion minutum', thinking it to be a sympathetic ganglion. Valentine, in 1840, saw a peculiar structure in the tympanic canaliculus in which a nerve cell was present and called it 'gangliolum tympanicum'. Almost 100 years later, in 1941, Stacey Guild reported the presence of some structures near the wall of the jugular bulb and described them as 'glomus jugulare'. Following this report, Rosenwaser (1945) and Otani were able to make a correct classification of these tumours in the middle ear. There have been innumerable reports of them and various authors have called them by various names. We report here a case of paraganglioma which was localized entirely to the tympanic membrane without involving any other part of the middle-ear cleft. PMID- 2987385 TI - Characterization of impact-induced tachyarrhythmia utilizing propranolol, quinidine and pacing. AB - Direct impact of the heart on the right ventricle induces transient ventricular tachycardia. Preconditioning the heart with a moderate dose of propranolol (0.4 mg/Kg, IV) did not affect impact-induced tachyarrhthmia although effective beta adrenergic blockage was evident. A high dose of propranolol (1.6 mg/Kg, IV) or a moderate dose of quinidine (3 mg/Kg, IV) prevented impact-induced ventricular tachycardia. High-dose propranolol and quinidine, an agent with no beta blockade properties, share similar local effects on the myocardial cell membrane which was sufficient to prevent the tachyarrhythmia. Pacing of the right ventricular free wall at the impact site resulted in an ECG similar to that observed following impact. Pacing of the subjacent septem, the other possible arrhythmogenic site affected by the impact, resulted in a large aberrant S wave (Lead II) different from the QRS complex during impact-induced tachyarrhythmia. Therefore, impact induced ventricular tachycardia originates from the impact site by a direct ventricular mechanism. PMID- 2987386 TI - Significance of ST-segment and T wave changes in the resting electrocardiograms of patients with exertional angina, studied by exercise radionuclide angiocardiograms. AB - In order to investigate the clinical significance of ST-T changes in resting ECG in angina pectoris, symptom-limited ergometer exercise radionuclide angiocardiography with ECG was performed in 60 patients with exertional angina. In those with normal ECG at rest (Group N), left ventricular ejection fraction (EF) did not change during exercise (71 +/- 5% to 71 +/- 6%). In those with only ST change at rest (Group ST) and those with only T change at rest (Group T), EF decreased significantly during exercise (68 +/- 5% to 63 +/- 7%, p less than 0.01; 68 +/- 6% to 61 +/- 7%, p less than 0.001). In those with ST and T changes at rest (Group ST + T), EF was low at rest (58 +/- 11%) and decreased further at exercise (52 +/- 8%, p less than 0.001). In those whose negative or flat T wave became more negative during exercise, EF was low both at rest and at exercise. In conclusion, ST and T changes at rest in patients with exertional angina might suggest a depressed reserve of myocardial function for exercise. PMID- 2987387 TI - Leydig cell resistance to the cytotoxic effect of ethylene dimethanesulphonate in the adult rat testis. AB - Weekly doses of the Leydig cell cytotoxic ethylene dimethanesulphonate (EDS) were administered to adult male rats in an attempt to study the endocrine activity of the testis in the absence of Leydig cells. One week after the first dose serum testosterone and LH concentrations and seminal vesicle weights were close to levels in castrated rats and testicular human chorionic gonadotrophin (hCG) binding was severely depressed. These changes were maintained for a further week but subsequently began to return to, but did not achieve, control levels. After six weekly doses seminal vesicle weight and serum testosterone concentrations were significantly higher than in the castrated rats. Serum LH concentrations were declining towards control values at 4 weeks but had risen again at 6 weeks. Serum FSH concentrations were raised to about 50% of the value in castrated rats throughout the period studied. Testis weight and hCG binding, which initially fell, were partially restored at 6 weeks and spermatogenesis was recovering. The data show that responses of the testis to multiple doses of EDS are similar to those after a single dose. This apparent resistance indicates that the regenerating Leydig cells are functionally different from the mature Leydig cell. The similarities between the maturing Leydig cell seen after EDS destruction and those in the immature rat suggest that EDS will provide a valuable model for the investigation of Leydig cell physiology. PMID- 2987388 TI - Antibiotics attenuate experimental hypertension in rats. AB - Hypertension was produced in Sprague-Dawley rats by intramuscular injections of either corticosterone or ACTH. Lower increases in blood pressure to these challenges were observed in Sprague-Dawley rats pretreated with neomycin or vancomycin which alone had no effect on blood pressure or growth. The development of high blood pressure in spontaneously hypertensive rats of a stroke-prone substrain was also attenuated by oral administration of neomycin. These results suggest that experimental hypertension can be modulated by the administration of antibiotics. PMID- 2987389 TI - Antagonistic actions of estradiol and tamoxifen upon forskolin-dependent meiotic arrest, intercellular coupling, and the cyclic AMP content of hamster oocyte cumulus complexes. AB - The effects of estradiol (E2) and the anti-E2 tamoxifen (tam) on forskolin (F) dependent meiotic arrest in hamster oocytes were investigated. The hypotheses tested were that 1) the arresting action of F is enhanced by E2 and suppressed by tam and 2) the extent of heterologous metabolic coupling and the concomitant transfer of cumulus cell cAMP into the oocyte is increased and decreased by E2 and tam, respectively. E2 was tested with the ID25 F (where ID25 is the dose of F previously shown to arrest the meiosis of 25% cultured oocytes; intact, 3 microM; denuded, 10 microM) and tam was tested with the ID75 F (intact, 10 microM; denuded, 100 microM). E2 induced reversible dose-dependent increases in the percent germinal vesicle (%GV; determined cytogenetically) of both intact and denuded oocytes in the presence of ID25 F (intact: ID50 E2 = 18.0 microM; denuded: ID50 E2 = 17.2 microM) but, in contrast to intact oocytes, E2 also exerted a dose-dependent action on denuded oocytes in the absence of F (ID50 = 26.1 microM). E2 induced dose-dependent increases in the cAMP content (determined by RIA) of intact oocytes (cAMP-oo) and of cumulus masses (cAMP-cm) and in the ratio of cAMPooo:cAMP-cm but failed to elevate F-stimulated cAMP in denuded oocytes. Heterologous metabolic coupling, as assessed by determination of the fraction of radiolabeled uridine marker that was transferred from the cumulus cells to the oocyte, was not significantly enhanced by E2. In contrast to denuded oocytes, tam induced dose-dependent decreases in the %GV and cAMP content of intact oocytes in the presence of ID75 F and significantly depressed heterologous metabolic coupling. While tam failed to antagonize the E2 action on denuded oocytes in the presence of ID25 F, in intact oocytes cultured with E2 and the ID25 F, the anti-E2 significantly decreased the %GV, the cAMP-oo and cAMP-cm, and the extent of heterologous metabolic coupling. These data show that while E2 can directly arrest the maturation of denuded hamster oocytes with no associated elevation of cAMP-oo, E2-enhancement of arrest in intact oocytes is correlated with both elevation of cAMP within the oocyte-cumulus complex and maintenance of heterologous metabolic coupling and is accompanied by an increase in cAMP-oo. PMID- 2987390 TI - cAMP in ovine oocytes: localization of synthesis and its action on protein synthesis, phosphorylation, and meiosis. AB - In the first series of experiments, the source of cAMP in the sheep oocyte was studied. Cholera toxin was shown to be a potent stimulator of cAMP in isolated sheep oocytes, demonstrating the presence of adenyl cyclase. There was no evidence for transmission of cAMP from stimulated myocardial cell monolayers to cumulus-enclosed oocytes even though the existence of a concentration gradient of cAMP and of intercellular communication were demonstrated. However, gonadotrophin stimulated follicle shells were able to induce a rise in the cAMP content of denuded or cumulus-enclosed oocytes in the same dish, independently of cell contact. Further experiments were designed to study the effects of a cholera toxin-stimulated rise in cAMP on the maturation of oocytes. When applied to cumulus-oocyte complexes, cholera toxin did not block germinal vesicle breakdown (GVBD), nor the accompanying changes in protein synthesis and phosphorylation, although there was evidence for a delaying effect. There were, however, indications that the toxin was inducing abnormalities that became gross when the concentration was raised to 1 microgram/ml. This high concentration of cholera toxin was able to block the maturation of oocytes in intact, gonadotrophin treated follicles, although once again abnormalities were evident. We conclude that the role of cAMP in the maturation of the sheep oocyte is different from that proposed in the mouse. PMID- 2987391 TI - [Subungual glomus tumor: histopathological, electron microscopic and immunohistochemical study on glomus tumor cells]. PMID- 2987393 TI - The role of polypeptides L and NS in the transcription process of vesicular stomatitis virus New Jersey using the temperature-sensitive mutant tsE1. AB - The roles of the L and NS polypeptides in transcription by vesicular stomatitis virus New Jersey were studied using a mutant, tsE1, which contains a temperature sensitive transcriptase and an altered NS polypeptide, both phenotypic changes being the consequence of the ts mutation. Mutant tsE1, its revertant (tsE1/R1) and the wild-type virus were dissociated into sub-viral fractions and, after reconstitution of these fractions in all combinations, the transcriptase was assayed in vitro at the permissive (31 degrees C) and restrictive (39 degrees C) temperatures. Reconstitution of the pellet fractions (containing polypeptide N complexed with the virion RNA) and the supernatant fractions (containing polypeptides L and NS) restored transcriptase activity at 31 degrees C in all combinations, but at 39 degrees C transcription was observed only in the presence of the supernatant fractions of wild-type and revertant viruses but not in the presence of the supernatant fractions of tsE1. When the pellet fractions and the L fractions were reconstituted, the transcriptase activity was restored in all combinations both at 31 degrees C and 39 degrees C. However, in vitro transcription at 39 degrees C by reconstituted pellet and L fractions was strongly inhibited when the NS fraction of tsE1 was also added, while addition of the NS fractions of wild-type and revertant viruses had no effect. Since only traces of polypeptide NS were present in the L fractions and none in the pellet fractions, the results strongly suggest that polypeptide L is the transcriptase itself while polypeptide NS exerts some control over transcription. PMID- 2987392 TI - Mechanism of osmotic activation of Na+/H+ exchange in rat thymic lymphocytes. AB - The activity of the Na+/H+ exchange system of rat thymic lymphocytes was determined by means of intracellular (pHi) and extracellular pH (pH0) measurements. In isotonic media, the antiport is virtually quiescent at physiological pHi (7.0-7.1), but is greatly activated by cytoplasmic acidification. At normal pHi, the antiport can also be activated by osmotic shrinking. Osmotic activation occurs after a delay of 20-30 s and is reversed several minutes after iso-osmolarity is restored. The mechanism of activation was analyzed by comparing the kinetic parameters of transport in resting (isotonic) and hyperosmotically stressed cells. The affinities of the external substrate site for Na+ and H+ are not altered in shrunken cells. In contrast, the Hi+ sensitivity of the antiport (which is largely dictated by an allosteric modifier site) was increased, which accounted for the activation. The concentration of free cytoplasmic Ca2+ [( Ca2+]i) increased after osmotic shrinking. This increase was dependent on the presence of extracellular Ca2+ and Na+ and was blocked by inhibitors of Na+/H+ exchange, which suggests that it is a consequence, rather than the cause, of the activation of the antiport. It is concluded that the shift in the pHi dependence of the modifier site of the Na+/H+ antiport is the primary event underlying the regulatory volume increase that follows osmotic shrinkage. PMID- 2987394 TI - In vitro phosphorylation of NS protein by the L protein of vesicular stomatitis virus. AB - The structural proteins L and NS of vesicular stomatitis virus were obtained from purified viral ribonucleoprotein complex followed by phosphocellulose column chromatography and assayed for protein kinase activity using [gamma-32P]ATP as the phosphate donor. The fractions containing purified L protein phosphorylated NS protein in vitro. 8-Azido-ATP, a photoreactive analogue of ATP, was also used as the phosphate donor for phosphorylation of NS protein by the L protein. In the presence of ultraviolet light, only L protein was specifically cross-linked with 8-azido-[gamma-32P]ATP. In the absence of u.v. light 8-azido ATP did no inhibit RNA transcription in a reconstituted reaction or substitute ATP for RNA synthesis in vitro. The above results, taken together, suggest that 8-azido-ATP was bound to the kinase site and phosphorylation of NS protein was mediated by the L protein. Exogenous phosphate acceptor proteins such as phosvitin and casein were also phosphorylated by the L protein fraction. However, addition of an excess of phosvitin failed to compete with the phosphorylation of NS by L, indicating that the protein kinase activity possessed higher affinity for NS. The phosphorylation of NS was strongly inhibited by photoreaction of L protein with 8-azido-ATP with concomitant inhibition of transcription in vitro. These results suggest that phosphorylation of NS protein by L may have a role in the regulation of the virus genome transcription in vitro. PMID- 2987396 TI - Introduction of cloned human papillomavirus 1a DNA into rat fibroblasts: integration, de novo methylation and absence of cellular morphological transformation. AB - The morphologically normal rat fibroblast cell line Rat-2 was used as a target cell type to test the transforming ability of a human papillomavirus (HPV-1a). To this end, molecularly cloned HPV-1a genomes were introduced into cultured Rat-2 cells in cotransfection experiments using a cloned herpes simplex virus thymidine kinase gene as a selectable phenotypic maker. In each of 13 HPV-1a-positive cell clones examined the papillomavirus DNA sequences were associated with the high molecular weight fraction of the cellular DNA, and restriction endonuclease plus Southern blotting analyses revealed patterns of hybridization which were consistent with integration of the viral genomes. Even Rat-2 clones containing multiple copies of the entire HPV-1a genome retained the normal, i.e. flat, cell morphology and were unable to grow in soft agar. De novo methylation of the HPV 1a sequences in many Rat-2 cell clones was evidenced by resistance of the viral DNA to complete cleavage with the HpaII endonuclease. Two out of three cell lines harbouring multiple copies of the HPV-1a genome contained detectable levels of HPV-1a transcripts, whereas no transcripts were detected in the third such cell line in which the viral HpaII sites were methylated virtually to completion. These results are consistent with the notion that HPV-1a genes are expressed inefficiently in Rat-2 cells; consequently integration of the viral DNA occurs, and there is no effect of the virus on the growth properties of this cell type. It is possible that methylation of the HPV-1a sequences is responsible for the low levels of expression of the viral genome. PMID- 2987395 TI - Preparation and characterization of neutralizing monoclonal antibodies with different reactivity patterns to human rotaviruses. AB - By employing three strains of cultivable human rotaviruses with different serotype specificity as immunizing antigens, we prepared 11 hybridomas which secreted neutralizing monoclonal antibodies against human rotaviruses. In neutralization tests with four strains of serotype 1, and three each of serotypes 2 and 3, the monoclonal antibodies showed different reactivity patterns: seven monoclonal antibodies reacted specifically with all strains of either serotype 1, 2 or 3 human rotavirus, but two showed strain-specific reactions; the remaining two were commonly reactive to various human rotavirus strains from each serotype but not to two non-human rotaviruses. By immunoprecipitation analysis, it was found that four serotype 2-specific and two commonly reactive antibodies were directed to VP3 (82000 mol. wt. protein) on the outer shell of the virus particles. PMID- 2987397 TI - Nuclear matrix modifications at different stages of infection by herpes simplex virus type 1. AB - In BHK-21 cells infected with herpes simplex virus type 1 many virus-induced proteins were found attached to the nuclear matrix. To understand the role of this cell fraction during virogenesis, matrix-associated proteins were analysed at different stages of infection. All the immediate-early protein species were bound to the nuclear matrix and their association with this structure was stable. During the first few hours of infection, the pattern of virus-induced proteins attached to the nuclear matrix remained identical, indicating that polypeptides from the early group are not associated with this cell fraction. Among the late proteins, which are generally structural proteins, 60% of the nuclear proteins were tightly bound to the nuclear matrix. This suggests that the nuclear matrix is involved in at least two different events during virogenesis, regulation of viral infection and assembly of viral capsids. PMID- 2987398 TI - Comparison of herpes simplex virus type 1 DNA replication and virus production in murine bone marrow-derived and resident peritoneal macrophages. AB - The mechanism of resistance of murine macrophages (M phi) to infection by herpes simplex virus type 1 (HSV-1) was examined. Infection of bone marrow-derived M phi (BMDM phi) and resident peritoneal M phi (Res-M phi) was compared with infection of permissive Vero cells. In contrast to HSV-1 infection in Vero cells, no infectious virus was produced from either M phi cell type. However, marked cytopathic effect (c.p.e.) was evident in BMDM phi at 48 h post-infection, while there was no c.p.e. at any time post-infection in the Res-M phi. Cloned EcoRI subgenomic fragments representing the entire HSV-1 genome were used as probes in DNA :DNA hybridization experiments to determine the viral genome content in the infected cell types. In Res-M phi, HSV-1 DNA was present at early times post infection but declined rapidly. In BMDM phi, the virus genome was always detected and increased with time after infection. The results suggest that Res-M phi restrict HSV-1 production at a point prior to viral DNA synthesis, whereas the block in HSV production in BMDM phi occurs at a later stage in the viral replicative cycle. PMID- 2987399 TI - Differences in the processing of secreted glycoprotein A induced by Marek's disease virus and herpesvirus of turkeys. AB - The steps from the precursor to the processed forms of a secreted glycoprotein (gA) of Marek's disease virus (MDV) and of herpesvirus of turkeys (HVT) were examined by two-dimensional gel electrophoresis of immunoprecipitates from the cell lysate and medium of infected cultures with monoclonal antibodies. Differences between MDV- and HVT-gA were particularly observed in isoelectric points of the glycosylated or unglycosylated precursor forms. PMID- 2987400 TI - Evidence that IFN-alph/beta induces two antiviral states active against different viruses. AB - IFN-alpha/beta has been suggested to require only one round of mRNA and protein synthesis to induce an antiviral state. We have examined the mechanism of induction of the antiviral states shown against three types of viruses: mengovirus (plus strand, sense RNA), vesicular stomatitis virus (VSV, minus strand RNA), and vaccinia virus (DNA). Mouse L cells were treated with IFN alpha/beta and cycloheximide and then with actinomycin D on a schedule which allowed only one round of mRNA and protein synthesis. The cells were challenged with virus under single cycle growth conditions to determine the amount of antiviral activity against the particular challenge virus employed. These studies confirmed that most of the antiviral effect directed against VSV is achieved with one round of macromolecular synthesis. However, most of the antiviral effect directed against mengovirus and vaccinia virus seemed to require more than one round. These results suggest that IFN-alpha/beta induces two different antiviral states: one requiring one round of synthesis which is primarily responsible for the inhibition observed for VSV; and another requiring more than one round of synthesis which is primarily responsible for the inhibition observed for mengovirus and vaccinia virus. PMID- 2987401 TI - MRC-5 cells, a model for Junin virus persistent infection. AB - Persistent infection of MRC-5 cells was established following inoculation with attenuated Junin virus (JV). In the acute phase of the infection both the pathogenic XJ and the attenuated XJ0 and XJC13 strains showed severe c.p.e. and free viral titres reached 10(5) p.f.u./ml. Recovery and establishment of persistently infected MRC-5 sublines (MRC-5PI) proved a very common event and seemed to be independent of viral strain, m.o.i. employed or virus passage history. These MRC-5PI sublines released virus throughout their life span and infectious centre assays performed at different passage levels with two sublines showed that 5 to 9% of the cells were producing virus. Heterotypic but not heterologous resistance to superinfection developed, as observed in persistent JV heteroploid cell systems. Analysis of released JV showed that attenuation had not been markedly altered, but alteration in plaque morphology under methyl cellulose, appearance of temperature-sensitive mutants and alterations in mouse pathology imply that some properties of JV have been altered. Results presented here stress once again the ability of JV to establish persistent infections. The source and diploid characteristics of MRC-5 cells make them a satisfactory model for JV persistence studies. PMID- 2987402 TI - Mouse mammary tumour virus: a proviral gene contributes to the understanding of eukaryotic gene expression and mammary tumorigenesis. PMID- 2987403 TI - Recognition of mos-related proteins with an antiserum to a peptide of the v-mos gene product. AB - A mos-specific antiserum was generated by injection of rabbits with a peptide predicted from the sequence of the v-mos gene of Moloney murine sarcoma virus (MuSV) strain 124. The peptide is composed of amino acids 37-55 (cyclized at the cysteine residues) conjugated to keyhole limpet haemocyanin. This serum [anti mos(37-55c)] specifically recognized p37mos in MuSV-124 acutely infected NIH-3T3 cells, P85gag-mos in 6m2 cells, an NRK clone infected with the temperature sensitive mutant (ts110) of Moloney MuSV, and P100gag-mos in 54-5A4 cells, an NRK clone infected with a spontaneous revertant of ts110. An additional protein of Mr 55000 from uninfected cells was recognized by this serum. Reactivity of the serum toward the v-mos-containing proteins and the 55K protein was completely inhibited by prior incubation with free peptide. The 55K protein was not recognized by antisera made from synthetic peptides prepared from the C-terminal eight or 12 amino acids of v-mos. PMID- 2987405 TI - Comparative studies of the binding of dimeric and monomeric enkephalins to neuroblastoma-glioma NG108-15 cells. AB - Binding activity of the enkephalin dimer [D-Ala2, Leu5-NH-CH2-]2 (DPE2) to NG108 15 hybrid cells was compared to that of the monomer [D-Ala2, Leu5]enkephalin amide (DALEA). At 25 degrees C, the values of the apparent affinity constant for DPE2, measured to intact and lysed cells and membranes, was 5.0 (+/- 0.09) X 10(9) M-1 for n = 28 experiments, as compared to 0.9 (+/- 0.08) X 10(9) M-1 (n = 16) for DALEA. At 4 degrees C, the binding affinity of DPE2 decreased by 43% and that of DALEA by 33%. An important difference between the binding of DPE2 and DALEA was that, after necessary corrections for difference in maximal "bindability" of the respective tritiated enkephalins, the molar binding capacity for DALEA was twofold higher than for DPE2, although mutual cross-displacement studies indicated that binding occurred to one class of noninteracting homogeneous receptors. The binding capacity for intact and lysed cells and membranes was 20 (+/- 2) X 10(-11) M for DPE2 and 43 (+/- 2) X 10(-11) M for DALEA. The enkephalin monomers [D-Ala2, D-Leu5]enkephalin (DADLE) and [D-Ala2, Met5]enkephalin amide (DAMEA) showed binding characteristics similar to those of DALEA. PMID- 2987404 TI - Three variations in the cell surface expression of the haemagglutinin neuraminidase glycoprotein of Sendai virus. AB - The fate of the haemagglutinin-neuraminidase glycoprotein (HN) of Sendai virus in three types of infection was studied by measuring its sensitivity to endoglycosidase H and its rate of appearance and turnover at the cell surface. HN behaved differently in the three types of infection. When highly expressed at the surface, as in a lytic standard virus infection, HN accumulated at the surface in a stable form (half-life of disappearance from the surface much greater than 10 h). When moderately expressed, as in a non-lytic standard virus plus defective interfering virus infection, HN reached the membrane normally, but turned over rapidly (half-life about 2 h) and was re-internalized. When poorly expressed, as in long-term persistent infection, HN did not reach the cell surface and appeared to be degraded before reaching it. In contrast to HN, the other viral glycoprotein, F0, exhibited a similar turnover rate at the cell surface in the three situations. However, when compared to surface expression in standard virus infected cells under standardized conditions, F0 surface expression in persistently infected cells was reduced. This reduction correlates with a decreased maturation rate in these cells. PMID- 2987406 TI - Radiation inactivation studies of the benzodiazepine/gamma-aminobutyric acid/chloride ionophore receptor complex. AB - Radiation inactivation was used to estimate the molecular weight of the benzodiazepine (BZ), gamma-aminobutyric acid (GABA), and associated chloride ionophore (picrotoxinin/barbiturate) binding sites in frozen membranes prepared from rat forebrain. The target size of the BZ recognition site (as defined by the binding of the agonists [3H]diazepam and [3H]flunitrazepam, the antagonists [3H]Ro 15-1788 and [3H]CGS 8216, and the inverse agonist [3H]ethyl-beta-carboline 3-carboxylate) averaged 51,000 +/- 2,000 daltons. The presence or absence of GABA during irradiation had no effect on the target size of the BZ recognition site. The apparent molecular weight of the GABA binding site labelled with [3H]muscimol was identical to the BZ receptor when determined under identical assay conditions. However the target size of the picrotoxinin/barbiturate binding site labelled with the cage convulsant [35S]t-butylbicyclophosphorothionate was about threefold larger (138,000 daltons). The effects of lyophilization on BZ receptor binding activity and target size analysis were also determined. A decrease in the number of BZ binding sites (Bmax) was observed in the nonirradiated, lyophilized membranes compared with frozen membranes. Lyophilization of membranes prior to irradiation at -135 degrees C or 30 degrees C resulted in a 53 and 151% increase, respectively, in the molecular weight (target size) estimates of the BZ recognition site when compared with frozen membrane preparations. Two enzymes were also added to the membrane preparations for subsequent target size analysis. In lyophilized preparations irradiated at 30 degrees C, the target size for beta galactosidase was also increased 71% when compared with frozen membrane preparations. In contrast, the target size for glucose-6-phosphate dehydrogenase was not altered by lyophilization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987407 TI - Calcium- and calmodulin-dependent phosphorylation of diphosphoinositide in acetylcholine receptor-rich membranes from electroplax of Narke japonica. AB - The phosphorylation of phosphoinositides in the acetylcholine receptor (AChR) rich membranes from the electroplax of the electric fish Narke japonica has been examined. When the AChR-rich membranes were incubated with [gamma-32P]ATP, 32P was incorporated into only two inositol phospholipids, i.e., tri- and diphosphoinositide (TPI and DPI). Even after the alkali treatment of the membrane, AChR-rich membranes still showed a considerable DPI kinase activity upon addition of exogenous DPI. It is likely that the 32P-incorporation into these lipids was realized by the membrane-bound DPI kinase and phosphatidyl inositol (PI) kinase. Such a membrane-bound DPI kinase was activated by Ca2+ (greater than 10(-6) M), whereas the PI kinase appeared to be inhibited by Ca2+. The effect of Ca2+ on the DPI phosphorylation was further enhanced by the addition of ubiquitous Ca2+-dependent regulator protein calmodulin. Calmodulin antagonists such as chlorpromazine (CPZ), trifluoperazine (TFP), and N-(6 aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) inhibited the phosphorylation of DPI in the AChR-rich membranes. It is suggested that the small pool of TPI in the plasma membrane is replenished by such Ca2+- and calmodulin-dependent DPI kinase responding to the change in the intracellular Ca2+ level. PMID- 2987408 TI - Acetylcholine receptors in small cell carcinomas. AB - The presence of muscarinic and nicotinic acetylcholine receptors (m- and nAChR, respectively) in small cell carcinomas (SCC) of the lung was assessed by measurement of specific binding of (-)[3H]quinuclidinyl benzilate ([3H]QNB) and 125I-alpha-bungarotoxin, respectively. Of five SCC studied, four were originally derived from patients with the Lambert-Eaton myasthenic syndrome, an autoimmune disease of neuromuscular transmission, and one was from a patient without evidence of neurological disease. There was no evidence of nAChR, but all tumors bound (-)[3H]QNB in a saturable and specific manner. Dissociation constants derived from saturation isotherms ranged between 35.4 and 181.7 fmol/mg protein (mean values for the five SCC). Competition studies revealed a pharmacological profile consistent with previous descriptions of mAChR. Competition by pirenzepine revealed only one class of binding sites, that with a relatively low affinity for pirenzepine. In the presence of the guanine nucleotide analogue 5' guanylyl imidodiphosphate, a decrease in affinity of the mAChR of SCC for oxotremorine was observed, with an increase in the pseudo-Hill coefficient, but there was no change in the binding of atropine. The expression on SCC of mAChR, apparently of the M2 subclass, represents yet another neural differentiation marker of SCC. It is noteworthy that the expression of this marker is not restricted to patients with an autoimmune paraneoplastic syndrome involving cholinergic neurons. PMID- 2987409 TI - Biphasic liberation of arachidonic and stearic acids during cerebral ischemia. AB - The mode of free fatty acid (FFA) liberation from the mouse brain during ischemia was investigated at various times after decapitation and under nizofenone treatment. Normal nonischemic brain FFAs consist mainly of palmitic acid (16:0), stearic acid (18:0), and oleic acid (18:1) with smaller amounts of arachidonic acid (20:4), docosahexaenoic acid (22:6), and others. Postdecapitative ischemia induced a rapid, biphasic release of 20:4 after a short lag of less than 30 s. The first phase showed a rapid 6.4-fold increase within 1 min of decapitation, followed by the second phase involving a slow release at less than one-fifth the rate of the first phase and lasting for at least 10 min. A similar, but not so marked, biphasic liberation was observed with 18:0. However, all of the other fatty acids (16:0, 18:1, 22:6, and others) were released only in a single phase at a slow rate. The time course for the rapid and specific liberation of 20:4 coincided with the time course for the decrease in brain ATP concentration during ischemia. Pretreatment of the animals with nizofenone resulted in a marked suppression of both FFA liberation and ATP depletion during ischemia. This suppression was particularly noteworthy with 20:4 and 18:0. The present study indicates that there is a specific and rapid liberation of 20:4 and 18:0 in a very early stage of ischemia and that this liberation seems to depend on availability of ATP in the brain. The physiological role of this transient 20:4 liberation during ischemia is discussed. PMID- 2987410 TI - High- and low-affinity binding components for [3H]imipramine in rat cerebral cortex. AB - The present study demonstrates that [3H]imipramine binds to both high- and low affinity imipramine binding components on membranes prepared from rat cerebral cortex. Scatchard and computer analyses of saturation experiments using a wide range of [3H]imipramine concentrations (0.5 nM-50 nM) revealed the presence of two binding components. Inhibition experiments in which membranes were incubated with [3H]imipramine and various concentrations of unlabelled imipramine gave shallow inhibition curves with a Hill coefficient of 0.60 +/- 0.04. When dissociation rates of imipramine were studied, biphasic dissociation curves were obtained with apparent half-times of dissociation of 2.5 +/- 0.4 min and 18.5 +/- 2.5 min. Thus analysis of saturation, competition, and dissociation experiments indicate that [3H]imipramine binds to low as well as high-affinity binding sites in rat cortex. PMID- 2987412 TI - Ca2+ and phospholipid-dependent protein kinase activity and phosphorylation of endogenous proteins in bovine adrenal medulla. AB - Soluble and membrane fractions of bovine adrenal medulla contain several substrates for the Ca2+/phospholipid-dependent and cyclic AMP-dependent protein kinases. The phosphorylation of soluble proteins (36 and 17.7 kilodaltons) and a membrane protein (22.5 kilodaltons) showed an absolute requirement for the presence of both Ca2+ and phosphatidylserine; other substrates showed less stringent phosphorylation requirements and many of these proteins were specific for each of the protein kinases. The Ca2+/phospholipid-dependent phosphorylation was rapid, with effects seen as early as at 30 s of incubation. Measurement of enzyme activities with histone H1 as an exogenous substrate demonstrated that the Ca2+/phospholipid-dependent protein kinase was equally distributed between the soluble and membrane fractions whereas the cyclic AMP-dependent enzyme was predominantly membrane-bound in adrenal medulla and chromaffin cells. The activity of the soluble Ca2+/phospholipid-dependent protein kinase of adrenal medulla was found to be about 50% of the enzyme level present in rat brain, a tissue previously shown to contain a very high enzyme activity. These results suggest a prominent role for the Ca2+/phospholipid-dependent protein kinase in chromaffin cell function. PMID- 2987413 TI - 5-Methoxytryptoline, a competitive endocoid acting at [3H]imipramine recognition sites in human platelets. AB - 5-Methoxytryptoline potently inhibits [3H]imipramine binding to membranes from the cerebral cortex and platelets. Since 5-methoxytryptoline, which appears to occur endogenously with particularly high levels in the human pineal gland, also inhibits 5-hydroxytryptamine (5-HT, serotonin) uptake, it should be considered as a putative endogenous ligand modulating 5-HT transport. As the 5-HT transporter complex comprises the imipramine and the substrate recognition sites, which interact allosterically, it was essential to define the mechanism of inhibition of [3H]imipramine binding by 5-methoxytryptoline. Human platelets show an active and saturable uptake of 5-HT and tryptamine. The uptake of both substrates appears to be mediated by the same carrier and it is inhibited by 5 methoxytryptoline at submicromolar concentrations. 5-HT and tryptamine inhibit [3H]imipramine binding in human platelets with a Hill slope for inhibition close to unity and IC50 values of 3,265 and 3,475 nM, respectively. This inhibition is, however, not competitive because both 5-HT and tryptamine significantly decrease the rate of [3H]imipramine-receptor dissociation. Although 5-methoxytryptoline potently inhibits [3H]imipramine binding (IC50 = 44 nM) in human platelets with a Hill slope of unity, it does not affect the receptor-ligand dissociation rate of [3H]imipramine even at concentrations up to 100 microM. The present experiments show that 5-methoxytryptoline, in spite of its chemical similarity to the indoleamine transporter substrates, interacts with the imipramine receptor through a mechanism of competitive inhibition. This conclusion is supported by a selective effect of 5-methoxytryptoline on the Kd of [3H]imipramine binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987411 TI - Differential degradation of different benzodiazepine binding proteins by incubation of membranes from cerebellum or hippocampus with trypsin. AB - When rat brain membranes were incubated with [3H]flunitrazepam in the presence of UV light, predominantly one protein (P51) was irreversibly labeled in cerebellum and at least two proteins (P51 and P55) were labeled in hippocampus. On digestion of membranes with increasing concentrations of trypsin up to 40% of radioactivity irreversibly bound to proteins was removed from the membranes. In addition, P51 was nearly completely degraded to a peptide with apparent molecular weight 39,000 and this peptide was further degraded to a peptide with apparent molecular weight 25,000. In contrast, protein P55 was only partially degraded by trypsin and yielded two proteolytic peptides with apparent molecular weights 42,000 and 45,000 which seemed to be rather stable against further attack by trypsin. Membranes treated with trypsin still had the capacity to bind [3H]-flunitrazepam reversibly with an affinity similar to that of membranes not previously treated with trypsin. When these membranes were irradiated with UV light, the same proteolytic peptides were detected as in membranes first photolabeled and then digested with trypsin. These results suggest a close association between reversible and irreversible benzodiazepine binding sites and indicate that membrane-associated proteins P51 and P55 are differentially protected against degradation by trypsin. PMID- 2987414 TI - Effects of taurine on calcium ion uptake and protein phosphorylation in rat retinal membrane preparations. AB - The effects of taurine on ATP-dependent calcium ion uptake and protein phosphorylation of rat retinal membrane preparations were investigated. Taurine (20 mM) stimulates ATP-dependent calcium ion uptake by twofold in crude retinal homogenates. In contrast, it inhibits the phosphorylation of specific membrane proteins as shown by acrylamide gel electrophoresis and autoradiography. The close structural analogue of taurine, 2-aminoethylhydrogen sulfate, demonstrates similar effects in both systems, i.e., stimulation of ATP-dependent calcium ion uptake and inhibition of protein phosphorylation, whereas isethionic acid and guanidinoethanesulfonate have no effect on either system. A P1 subcellular fraction of the retinal membrane preparation that contains photoreceptor cell synaptosomes has a higher specific activity for the uptake of calcium ions. Phosphorylation of specific proteins in the P1 fraction is also inhibited by the addition of 20 mM taurine. Taurine has no effect on retinal ATPase activities or on phosphatase activity, thus suggesting that it directly affects a kinase system. PMID- 2987415 TI - Dephosphorylation of microtubule-associated protein 2, tau factor, and tubulin by calcineurin. AB - Calcineurin dephosphorylated microtubule-associated protein 2 (MAP2) and tau factor phosphorylated by cyclic AMP-dependent and Ca2+, calmodulin-dependent protein kinases from the brain. Tubulin, only phosphorylated by the Ca2+, calmodulin-dependent protein kinase, served as substrate for calcineurin. The concentrations of calmodulin required to give half-maximal activation of calcineurin were 21 and 16 nM with MAP2 and tau factor as substrates, respectively. The Km and Vmax values were in ranges of 1-3 microM and 0.4-1.7 mumol/mg/min, respectively, for MAP2 and tau factor. The Km value for tubulin was in a similar range, but the Vmax value was lower. The peptide map analysis revealed that calcineurin dephosphorylated MAP2 and tau factor universally, but not in a site-specific manner. The autophosphorylated Ca2+, calmodulin-dependent protein kinase was not dephosphorylated by calcineurin. These results suggest that calcineurin plays an important role in the functions of microtubules via dephosphorylation. PMID- 2987416 TI - Biochemical studies on 5'-nucleotidase of Schwann cells in degenerated nerve. AB - This report describes the partial characterization of 5'-nucleotidase (5'-AMPase) in Schwann-cell plasmalemmae (PM) prepared from degenerated cat sciatic nerve. 5' AMPase was enriched 3.7-fold in the PM fraction over that of the crude homogenate preparation. The plant lectin concanavalin-A (Con-A) reduced Schwann cell PM 5' AMPase activity in a concentration-dependent manner (30-600 micrograms/ml). Plasma membrane 5'-AMPase activity was maximally inhibited to 20% of control values by Con-A (400-600 micrograms/ml), and activity returned to control levels by pretreatment with the hapten sugar alpha-methyl-D-mannoside (50 mM). Equimolar concentrations of UDP and ADP (100 microM) reduced the rate of hydrolysis of labeled AMP to labeled adenosine in PM to 45% and 35% of control, respectively. This is the first study to characterize a Schwann-cell PM enzyme and demonstrates that 5'-AMPase may be used as a Schwann-cell PM marker enzyme. PMID- 2987417 TI - Glucocorticoids potentiate the prostaglandin E1-mediated cyclic AMP formation by a cultured murine neuroblastoma clone. AB - The effect of steroid hormones on the prostaglandin E1 (PGE1)-mediated cyclic AMP formation by murine neuroblastoma clone N1E-115 was studied. Dexamethasone at submicromolar concentrations and corticosterone at micromolar concentrations (steroids with glucocorticoid activity) were able to modify the PGE1-mediated response whereas testosterone, progesterone, and estradiol each at 10 microM had no effect. Glucocorticoids added to the culture medium of N1E-115 cells produced an increase in the maximal response to PGE1 only after long-term (greater than or equal to 4 h) incubation with the hormone. Inhibitors of protein and RNA synthesis blocked this effect of glucocorticoids. Basal activity of adenylate cyclase in treated cells was twofold higher than that in control cells, and this enzyme seemed to be the primary target for the hormone action, since the activity of 3':5'-cyclic AMP phosphodiesterase and the binding of [3H]PGE1 to its receptors were not altered by glucocorticoid treatment. Our results indicate that glucocorticoids modulate receptor-mediated responses in cells of neural origin through a mechanism that involves induction of protein synthesis. PMID- 2987418 TI - Blood flow and oxygen utilisation in the contralateral cerebral cortex of patients with untreated intracranial tumours as studied by positron emission tomography, with observations on the effect of decompressive surgery. AB - Using positron emission tomography, regional cerebral blood flow and oxygen utilisation were measured in the contralateral cortex of 14 patients with a variety of intracranial tumours. A comparison was made with cortical values derived from 14 normal controls. Compared with normal subjects, patients with brain tumours had a significant reduction in oxygen utilisation and blood flow in their contralateral cortex. Decompression resulting from craniotomy and biopsy, led to a partial reversal of this cerebral hypofunction. PMID- 2987420 TI - Sympathetic skin response. PMID- 2987419 TI - An improved automated method for the measurement of thermal thresholds. 2. Patients with peripheral neuropathy. AB - Thermal thresholds were determined by a new technique, at wrists and ankles in 143 patients with peripheral neuropathies of diverse aetiologies. Ninety-nine percent of patients (141/143) had abnormalities of one or both thresholds. In only two patients with mild/early Friedreich's ataxia were thermal thresholds normal. Electromyography was performed and fastest motor nerve conduction velocities and sensory nerve action potential parameters were measured in all the patients using conventional techniques in ulnar, median and sural nerves. Eighty nine percent of patients (127/143) had one or more abnormalities on these electrophysiological studies. However, 39 of 40 patients with completely normal sensory nerve studies had an abnormality of one or more thermal thresholds. Eighty-six percent of 48 patients with normal sural nerve studies had abnormal thermal thresholds at the ankle. Sixty percent of 70 patients with normal sensory median and ulnar nerve studies had abnormal wrist thermal thresholds. This improved technique for the determination of thermal thresholds reveals that disturbances of thermal sensibility are present in the majority of peripheral neuropathies irrespective of aetiology. In some patients disturbances of thermal thresholds antedate the appearance of abnormalities on conventional electrophysiological investigation. The findings suggest that this technique has considerable usefulness in the detection of small nerve fibre dysfunction in the context of generalised neuropathy. PMID- 2987421 TI - Production of and response to interleukin-2 by cultured T cells: effects of lithium chloride and other putative modulators. AB - Immunomodulating substances may be useful as host-modifying adjuvants in patients receiving immunosuppressive therapies or as probes for studying the immune system. Detection of interleukin-2 (IL-2) is possible via a microassay, enabling investigation of the effects of immunomodulating agents on T cell production of and response to IL-2 in vitro. Four such agents were examined: lithium chloride, Bestatin, Azimexon, and muramyl dipeptide (MDP). All four agents increased the response of cultured T cells to IL-2 over control populations supplied with IL-2 alone. Lithium chloride, Bestatin, and MDP enhanced the production of IL-2 in standard mixed lymphocyte cultures over control cultures receiving media alone. These results suggest that several immunomodulators may produce their effects by the modulation of IL-2 or other lymphokines. Additionally, microassay systems may prove useful as probes to elucidate the function of putative immunoregulatory substances. PMID- 2987422 TI - Superoxide anion (O-2) production by peripheral blood monocytes in Hodgkin's disease and malignant lymphoma. AB - Superoxide anion (O-2) is the first metabolite of the monocyte oxygen burst pathway, which plays an important role in the monocyte microbicidal function. The capacity of peripheral blood monocytes to produce O-2 was studied in 63 patients with Hodgkin's disease (31 with active disease and 32 in complete remission), 15 patients with active malignant lymphoma, and 57 normal control subjects. O-2 release was quantified by evaluating superoxide dismutase-inhibitable reduction of cytochrome c after stimulation of monocytes with phorbol myristate acetate. Results were expressed in nanomols O-2 per mg protein per hour. O-2 production was lower than normal in patients with active Hodgkin's disease (163.3 v 214.5, P less than .05). It was normal in patients with Hodgkin's disease in complete remission (216.2 v 214.5, P greater than .05) and high in patients with malignant lymphomas (317.9 v 214.5, P less than .01). Within the group with active Hodgkin's disease, patients in relapse after therapy had a lower O-2 production than those previously untreated (99.8 v 181.8, P less than .01). Stage of disease was unrelated to the defect. The presence of B symptoms and a decreased delayed type hypersensitivity to recall skin test antigens were associated with normal O 2 production. The results obtained suggest that monocyte dysfunction is part of the immune dysregulation associated with active Hodgkin's disease. The O-2 determination is a relatively easy test to perform and may be useful in identifying the patients with Hodgkin's disease who have an increased risk of opportunistic infections. PMID- 2987423 TI - Radiotherapy and combination chemotherapy with carmustine, vincristine, and procarbazine (BVP) in primary brain tumors. AB - 26 patients with astrocytoma grade II-III, and 36 with malignant glioma (astrocytoma grade IV or glioblastoma) were submitted three days after surgery to a cycle of combination chemotherapy, including BCNU, VCR, PCZ (BVP). Eighteen days after surgery, patients received 40 Gy (astrocytoma grade II-III) or 45 Gy (malignant glioma) of megavoltage whole-brain irradiation, with an additional boost to the 'tumor' bed of 20 Gy, delivered in 6 weeks. Vincristine was injected weekly during radiotherapy. At the end of radiotherapy, patients received BVP every 6 weeks for at least 8 cycles or until a recurrence or progressive disease. Performance status of grade 1 or 2 was achieved in 15 (60%) and in 5 (20%), respectively, of patients with astrocytoma grade II-III after 6 months, and in 6 ps. (29%) and in 9 ps. (42%) after 12 months of follow-up. Only 2 (5.5%) and 18 (64%) patients with malignant glioma achieved a performance status of grade 1 or 2 after 6 months, and these proportions are 6% and 35%, respectively, after 12 months. After a 5-year follow-up, 59% of patients with astrocytoma are still alive, with a median survival time of 60+ months, whereas only 4% of patients with malignant glioma are alive, with a median of 11.2 months. PMID- 2987425 TI - Primitive neuroectodermal tumours of the cerebrum. Pathology and treatment. AB - Eighteen cases of cerebral tumour composed partly or totally of primitive embryonal cells are reported. These lesions comprise 2.8% of all primary cerebral hemisphere tumours in the histopathology files of The Royal Marsden Hospital between 1971 and 1980 inclusive. Most exhibited some degree of differentiation towards neuronal or glial elements and, as more than one type of differentiation was often present in the same lesion, we agree with others that the term primitive neuroectodermal tumour (PNET) is more appropriate to describe these lesions than terms based on histogenesis. The extent of the primitive component varied, but usually accounted for more than 80% of the tumour. Although the tumours bear some similarities to posterior fossa medulloblastomas, they exhibit important differences in histology, immunohistology, natural history and response to treatment. Nearly all PNETs examined expressed some glial fibrillary acidic (GFAP) both in primitive areas and zones of astrocytic differentiation. GFAP staining may thus be of value in distinguishing PNETs from undifferentiated non neurogenic tumours. Of 14 patients referred for radiotherapy, the survival rate at 3 years was 29% (4/14) and 5 years 25% (3/12). Patients with tumours in which at least 90% of the tissue was undifferentiated exhibited an extremely poor prognosis with none of 9 patients still alive at 3 years in contrast to 3 of 5 patients (60%) with tumours showing less than 90% undifferentiation. Radical tumour removal, where feasible, followed by irradiation of the whole cerebrospinal axis is recommended. Adjuvant chemotherapy with such agents as CCNU and Vincristine may be of value: the 3 long term survivors in the present series (7-11 years), including one who presented disseminated intracranial disease, received such adjuvant treatment. PMID- 2987424 TI - Phase II study of aziridinylbenzoquinone (AZQ: NSC-182986) in the treatment of malignant gliomas recurrent after radiation. Preliminary report. AB - Seventeen patients with malignant gliomas recurrent after chemotherapy and/or radiation failure were treated with aziridinylbenzoquinone (AZQ) at a dose of 20 15 mg/M2 weekly for four weeks followed by a two week rest. Regression of disease was observed in four patients, 4/17 (24%) for 35, 15+, 40+, and 10 weeks. Toxicity was limited to moderate reversible myelosuppression. AZQ in this dose and schedule has limited but definite activity in patients with malignant gliomas progressive after primary radiation therapy failure. PMID- 2987426 TI - Human neuroblastoma cells and 13-cis-retinoic acid. AB - To a limited extent, we corroborated a previous report of human neuroblastoma sensitivity to 13-cis-retinoic acid. Seven cultured human neuroblastoma, two primitive neuroectodermal tumor, and one melanoma cell line were exposed to 0.001 to 10.0 microM 13-cis-retinoic acid for six to fourteen days. The neuroblastoma cell line, SK-N-DZ, was the only cell line lysed by all concentrations of 13-cis retinoic acid. The other cell lines were refractory to concentrations as high as 10 microM. Increased cell process formation was observed in three neuroblastoma, SK-N-SH, SK-N-BE, SK-N-LE, and one melanoma cell line. We conclude that sensitivity to 13-cis-retinoic acid is unevenly distributed among histogenetically similar tumors from different patients. PMID- 2987427 TI - Supraophthalmic carotid infusion for recurrent glioma: rationale, technique, and preliminary results for cisplatin and BCNU. AB - The chemotherapeutic agents 1-3 bis(2-chloroethyl)-1-nitrosourea (BCNU) and cis diamminedichloroplatinum II (cisplatin) have both shown activity against malignant glioma, especially when given by arterial infusion. The combination of these agents given by this method is logical because their individual major toxicities are directed at different organ systems, and because of their differences in restriction by the blood-brain barrier. Both agents are toxic to the eye, and infusion of both agents simultaneously into the internal carotid artery would deliver doses of the drug to the eye which should be associated with an unacceptable level of ocular toxicity. We have developed a technique utilizing a flexible flow-directed catheter with a tip which is manipulated by hydraulic forces for delivery of the drug into the intracranial carotid artery above the origin of the ophthalmic artery, thus sparing the eye from the high concentration of drug during the first pass through the arterial circulation. In 13 patients with recurrent malignant glioma treated by arterial infusion of both agents (cisplatin 150-200 mg, BCNU 300 mg fixed dose), we have had no damage to the ipsilateral eye. Preliminary results of treatment appear to be good, with definite tumor regression following arterial infusion in 10 of 12 radiographically evaluable cases. Median survival to date is 11 months with 3 patients still surviving. The longest survival is 24 months. The supraophthalmic infusion technique protects the eye and the combination of drugs given by arterial infusion produces a high tumor response rate. PMID- 2987428 TI - Immunohistochemical study of glial fibrillary acidic protein in avian sarcoma virus-induced gliomas in dogs. AB - Anaplastic astrocytomas, gliosarcomas and sarcomas were induced in 33 neonatal dogs by intracerebral injection of purified Schmidt-Ruppin strain of avian sarcoma virus (ASV). A total of 11 anaplastic gliomas, 7 gliosarcomas and 18 sarcomas were induced. Ten of the 11 anaplastic astrocytomas and all 7 gliosarcomas were positive for GFAP using the peroxidase-antiperoxidase method on formalin-fixed, paraffin embedded tissue. None of the sarcomas were positive for GFAP. One anaplastic astrocytoma was analyzed by gel electrophoresis and immunoblot and the amount of GFAP in the tumor was compared with comparable samples from normal white and gray matter. Densitometric analysis of the GFAP stained gels showed that 12-13% of tumor protein was GFAP compared with 2-3% for the non-neoplastic white and gray matter. The results of this study add further evidence to the astrocytic origin of many of the induced anaplastic primary brain tumors by ASV. PMID- 2987429 TI - Phase I clinical trial of intralesional or intraventricular leukocyte interferon for intracranial malignancies. AB - Eight patients were treated with leukocyte interferon for a variety of neurological malignancies that had failed or recurred after conventional therapy. Three patients with malignant astrocytoma received intratumoral interferon in dosages up to 9 million units 3X/week, with total dosages of up to 160 million units. Interferon was administered intraventricularly in 4 patients with leptomeningeal metastases and one patient with multiple brain metastases. Dosages increased from 1 to 10 million units 3X/week, and total dosages of up to 113 million units were given intraventricularly. Acute side effects of fever, nausea, vomiting, and headache occurred almost exclusively with intraventricular injections, and these subsided after the initial injection. Fatigue, loss of appetite, weight loss, and hematologic toxicity developed a few weeks after onset of treatment, independent of the dose given. A modest tumor regression was seen on CT scans of one patient with a malignant astrocytoma, who was treated with interferon for 8 months. In all 4 patients with leptomeningeal metastases, the CSF became free of malignant cells for 6 to 10 weeks, while clinical improvement was less dramatic. PMID- 2987430 TI - Metrizamide myelography in patients with small cell carcinoma of the lung suspected of meningeal carcinomatosis. AB - Twenty patients suspected of meningeal carcinomatosis (MC) secondary to small cell carcinoma of the lung were evaluated by myelography and by cytologic examination of cerebrospinal fluid (CSF). In 4 patients both investigations demonstrated MC. Four had positive cytology but a negative myelogram. Five patients had a positive myelogram but negative cytology. Thus 13 of the 20 patients had MC diagnosed by one or both of these investigations. Two additional patients suspected of MC were studied; both had positive myelography, but cytologic examination of CSF was not performed in either case. Six patients underwent autopsy with microscopic examination of the spinal cord; in all cases post-mortem findings supported myelographic findings. In conclusion, myelography is useful in the evaluation of patients suspected of MC secondary to small carcinoma of the lung and complements the diagnostic role of CSF examination. PMID- 2987431 TI - Histaminergic synaptic transmission in the cerebral ganglion of Aplysia. AB - Standard intracellular stimulating and recording techniques including voltage clamp were used to analyze the synaptic responses mediated by two identifiable histamine-containing neurons (HCNs), designated C2 neurons, located in bilaterally symmetric clusters of the isolated cerebral ganglion of Aplysia california. Activation of each C2 induced unitary chemically mediated synaptic potentials in over 15 identified ipsilateral follower neurons. Several additional followers were connected to the HCNs by nonrectifying electrical synapses. Most of the follower neurons examined received only chemical synapses from the C2s. Some of the followers were reciprocally connected with each other through nonrectifying electrical synapses. A single C2 action potential can evoke six distinctive types of chemically mediated postsynaptic potentials (PSPs) in different follower neurons. Most of the PSPs have been shown to be multicomponent, i.e., they are comprised of various combinations of individual fast (less than or equal to 150 ms), slow (1-2 s), and very slow (greater than or equal to 4 s) depolarizing and hyperpolarizing components. The combination of these components produces PSPs of varying complexity, from simple monophasic responses such as the frequently observed slow excitatory PSPs and slow inhibitory PSPs to responses consisting of two to three components such as fast excitatory, slow inhibitory PSPs or fast inhibitory, slow excitatory PSPs. All of the multicomponent PSPs appear to be mediated through monosynaptic connections from the C2, as determined by various electrophysiological criteria. The slow and very slow synaptic components of the multicomponent PSPs were markedly potentiated in amplitude and duration after repetitive C2 activation. This property of the slow components permits the slower PSPs to exert a major influence on the excitability and integrative properties of the follower neurons. PMID- 2987432 TI - Integration of nonphaselocked exteroceptive information in the control of rhythmic flight in the locust. AB - The integration of exteroceptive information in the flight control system of the locust was studied by determining the cellular basis of ocellar- (simple eye) mediated control of flight. Neural interactions that transform phase-independent sensory input into phase-specific motor output were characterized. Ocellar information about course deviations during flight was conveyed to the segmental thoracic ganglia by three pairs of large fast multimodal descending neurons. These made connections with thoracic motoneurons directly, via short-latency mono or disynaptic pathways, and indirectly, via a population of intercalated thoracic interneurons. The synaptic potentials caused in the motoneurons by the direct pathway occurred at short latency and were adequate for summation with other types of sensory input. However, the strength of the synaptic effects of this pathway was weak compared with the central flight oscillator drive to the same motoneurons. In contrast, synaptic potentials evoked by the descending neurons in the thoracic interneurons were often large and brought these cells close to threshold. In turn, these interneurons always had stronger synaptic effects on postsynaptic flight motoneurons than did the descending neurons alone. We conclude that the indirect interneuronal pathway is more powerful in its effects on motoneurons than the direct pathway. Premotor thoracic interneurons, which received ocellar input appropriate for a role in correctional steering, were also rhythmically modulated during flight motor activity in phase with either depressor or elevator motoneurons. This phasic modulatory drive occurred in deafferented preparations, indicating that its source is the central oscillator for flight. Presentation of ocellar stimulation during flight motor activity showed that the central oscillatory modulation of the thoracic interneurons gated the transmission of sensory information through these interneurons. Ocellar mediated postsynaptic potentials influenced the firing of thoracic interneurons only if they arrived during the proper phase of rhythmic drive. Thus the transmission of ocellar information from interneuron to motor neuron is possible only during appropriate phases of the flight cycle. PMID- 2987433 TI - Relationship among recruitment order, axonal conduction velocity, and muscle-unit properties of type-identified motor units in cat plantaris muscle. AB - A strict interpretation of the size-principle hypothesis (37, 39-41) is that a muscle's motor units should be recruited in an ascending order according to both the size of their motoneurons and the size of their innervated muscle units (for reviews see Refs. 9, 39, 73). Studies of large mixed muscles in the cat hindlimb, however, have shown that motor axonal conduction velocity and tetanic tension, which are frequently considered indices of motoneuron and muscle-unit size, respectively, are uncorrelated in the fast-twitch (type F) motor-unit subpopulation (12, 13, 23, 24, 30, 32, 63, 71, 79). Attempting to focus on type F units, we compared the recruitment order of 42 pairs of cat plantaris (PL) motor units with both axonal conduction velocity and tetanic tension as well as with other muscle-unit properties. Single PL alpha-motor axons were functionally isolated in intact L7 ventral root filaments of decerebrate cats. Tension responses produced by stimulating each isolated motor axon were used to find the tetanic tension of the muscle unit and to classify the unit (12) as either type S (slow twitch, fatigue resistant), type FR (fast twitch, fatigue resistant), type FI (fast twitch, intermediate fatigability), or type FF (fast twitch, highly fatigable). Conduction velocity of each isolated axon was computed from measurements of axonal conduction time and length. The axon's reflex discharges were recorded from the proximal end of the cut filament and compared with the discharges of another PL axon residing in a different, previously cut filament of the same cat. The recruitment order of each motor-unit pair studied was found during reflexes elicited by homonymous muscle stretch, tendon taps, or single shocks at group I intensity to the PL nerve. If either axon of the pair failed to discharge, as often was the case with the high-threshold type F units, the monosynaptic reflex was facilitated by a 500-pps conditioning train applied proximal to a complete reversible cooling block of the PL nerve. In all 42 pairs studied, the weaker motor unit had the lower functional threshold for recruitment. Recruitment also invariably followed the order S greater than FR greater than FI greater than FF units. The motor unit of a pair with the higher resistance to fatigue thus always had the lower functional threshold. In 21 of the 22 pairs containing at least one type S motor unit, the unit with the slower conducting motor axon had the lower functional threshold for recruitment.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2987434 TI - Nerve growth factor treatment enhances nicotine-stimulated dopamine release and increases in cyclic adenosine 3':5'-monophosphate levels in PC12 cell cultures. AB - In order to examine the relationship between cyclic AMP (cAMP) levels and evoked neurotransmitter release, experiments have been performed with cultures of clonal rat PC12 pheochromocytoma cells. Stimulation of the release of endogenous dopamine by nicotine in these cultures is calcium-dependent and blocked by d tubocurarine, a specific nicotinic cholinergic antagonist. Similarly, nicotine causes increases in cAMP levels in PC12 cell cultures that are calcium-dependent and blocked by d-tubocurarine. Cultures treated for 6 days or longer with 2 X 10( 9) M nerve growth factor (NGF) release a 3- to 4-fold greater amount of dopamine than do control cultures in response to a maximal concentration of nicotine. Correspondingly, nicotine causes a 3-fold greater increase in cAMP levels in the NGF-treated cultures than in the controls. These results suggest that stimulation of the nicotinic cholinergic receptor in PC12 cells results in some manner in the activation of adenylate cyclase and further support the notion that cAMP is involved in the process of neurotransmitter release. PMID- 2987435 TI - Regulation of synthesis of the neurosecretory egg-laying hormone of Aplysia: antagonistic roles of calcium and cyclic adenosine 3':5'-monophosphate. AB - The potential role of cyclic nucleotides and calcium as regulators of neuropeptide biosynthesis was examined in the bag cell neurons of Aplysia, which produce and secrete a peptide egg-laying hormone (ELH). Elevated external potassium, which stimulates ELH biosynthesis, increased bag cell cAMP levels when assayed in the presence of a phosphodiesterase inhibitor. Dopamine and serotonin, which increase bag cell cAMP levels, both stimulated ELH synthesis, as did the phosphodiesterase inhibitor isobutylmethylxanthine, the specific adenylate cyclase activator forskolin, and the phosphodiesterase-resistant cAMP analogue 8 benzylthio-cAMP. The stimulatory effect on peptide biosynthesis appears to be specific for cAMP, as bag cell cGMP levels were not altered significantly by high potassium or forskolin, and 8-bromo-cGMP did not stimulate ELH synthesis. In contrast to cAMP, intracellular calcium inhibits ELH production: biosynthesis of the peptide was elevated in a 0 Ca2+/EGTA medium and reduced in the presence of the Ca2+ ionophore A23187. Synthesis was also elevated in the presence of the calmodulin inhibitor calmidazolium. Treatment of intact bag cells with 0 Ca2+/EGTA or A23187 did not alter cAMP levels significantly, suggesting that calcium exerts its effect on peptide synthesis independently of cAMP. The antagonistic effects of cAMP and calcium on ELH synthesis parallel their effects on bag cell excitability, suggesting that, in these cells, neuropeptide synthesis and secretion are co-regulated by the same intracellular messengers. PMID- 2987436 TI - Glutamic acid decarboxylase in the striate cortex of normal and monocularly deprived kittens. AB - Degeneration of the thalamic fibers in the visual cortex of turtles leads to an increase in the numerical density of cortical synapses with flattened vesicles and symmetrical membrane differentiations (Smith, L. M., and F. F. Ebner (1980) Soc. Neurosci. Abstr. 6: 328). This change correlates with an increase in the cortical activity of glutamic acid decarboxylase (GAD), the synthetic enzyme for gamma-aminobutyric acid (GABA). These data are consistent with the hypothesis that removal of thalamic input activity is the stimulus for cortical GABAergic neurons to form new synapses. Pharmacological evidence suggests that even simple environmental deprivation may induce a similar increase in the numerical density of GABAergic synapses in kitten striate cortex (Duffy, F. H., S. R., Snodgrass, J. L. Burchfiel, and J. L. Conway (1976) Nature 260: 256-257). We have examined this possibility in monocularly deprived kittens using methods to localize and measure GAD. GAD in kitten striate cortex was localized using immunocytochemistry. GAD-positive cells were found in all layers and were uniformly distributed in layers II to VI. Immunoreactivity associated with axon terminals (puncta), in contrast, was laminated with a distinct band in layer IV. Monocular deprivation (MD), by either unilateral enucleation or lid closure, had no detectable effect on the distribution of GAD in striate cortex. The band of layer IV puncta remained uniform even under conditions that produced alterations in layer IV cytochrome oxidase activity. We measured GAD activity in homogenates of striate cortex to address the possibility that MD causes an absolute change in the density of GABAergic synapses. Again, however, GAD activity in the binocular and monocular segments of striate cortex was found to be unaffected by early enucleation. These data suggest two conclusions: first, that the numerical density of GABAergic synapses in visual cortex is not regulated directly by thalamic activity, and second, that changes in GABAergic synapse density do not account for the ocular dominance shift observed in kitten striate cortex after MD. PMID- 2987437 TI - Morphological and functional types of neurons in cat ventral posterior thalamic nucleus. AB - Neurons in the thalamic ventral posterior (VB) nucleus of the cat were investigated by extracellular and intracellular recording and by anatomical methods involving either the retrograde transport of horseradish peroxidase (HRP) or the intracellular injection of HRP. Two morphological types of neurons could be detected by retrograde labeling from small injections of HRP in the internal capsule adjacent to VB. These two and one other type, judged to be an interneuron, could also be identified by intracellular staining. Type I cells are large, have thick proximal dendrites which branch in a tuft-like manner, and thick, rapidly conducting axons. They possess few or no dendritic appendages. Type II cells are smaller and have slender proximal dendrites which branch dichotomously and thin, slower conducting axons. Those injected intracellularly are covered in fine, hair-like dendritic appendages. Type III cells are small and have thin processes that give rise to many bulbous dilatations and no obvious axon. Type I and type II cells give off slender axon collaterals in the thalamic reticular nucleus but not in VB. Examples of both types of cell could be antidromically activated from the somatic sensory cortex. Type I and type II cells recovered histologically after intracellular recording included examples of most types of receptive field, including several forms of cutaneous and deep fields, as classified by us in a parallel intra- and extracellular study of unit responses. All but one type I cell, however, responded in a transient manner to peripheral stimulation. The remaining type I cell and all members of an admittedly small sample of type II cells responded in a sustained manner. The sample of recovered interneurons and of units that could not be driven antidromically from the cerebral cortex suggested that they, too, included all receptive field types. We conclude that submodality specificity in VB is not represented by morphological specificity in thalamocortical relay cells or interneurons. Some other functional parameter, such as tonic or phasic responsiveness, may be more obviously correlated with relay cell morphology. PMID- 2987438 TI - Peptidergic neurons of Aplysia lose their response to cyclic adenosine 3':5' monophosphate during a prolonged refractory period. AB - Although the peptidergic bag cell neurons of Aplysia are ordinarily silent, they respond to brief electrical stimulation by producing an afterdischarge of about 30 min duration. This afterdischarge is followed by a refractory period lasting many hours during which electrical stimulation either fails to initiate afterdischarges or produces discharges of much shorter duration. Previous work has demonstrated that cyclic AMP plays a role in the genesis of afterdischarge, both in intact bag cell clusters and in isolated cultured bag cells. We have now examined the hypothesis that in the refractory period either the synthesis of cyclic AMP or the response to cyclic AMP is attenuated. Direct measurements of cyclic AMP showed that cyclic AMP levels in the bag cell neurons are elevated to a similar extent after stimulation in refractory and nonrefractory clusters of neurons. We have found, however, that the response to cyclic AMP is altered during the refractory period. The electrophysiological responses of bag cell neurons were first examined in intact clusters of cells within the abdominal ganglion. Cyclic AMP levels were elevated using the adenylate cyclase activator, forskolin, in the presence of theophylline (FT). The duration of a first bag cell afterdischarge could be greatly increased if FT was added before stimulation. The duration of a stimulated second bag cell afterdischarge could also be significantly increased if FT was added within a brief period following the end of the first afterdischarge. Furthermore, at these times the addition of FT often resulted in the onset of spontaneous afterdischarges.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987439 TI - Embryonal central neuroepithelial tumors and their differentiating potential. A cytogenetic view of a complex neuro-oncological problem. AB - The embryonal central nervous system (CNS) neoplasms are reviewed with special reference to their differentiating potential and in the light of current neuro oncogenetic concepts partly derived from the experimental induction of neural tumors. The conceptual (and, occasionally, practical) distinction between adult type and embryonal CNS tumors raises a complex problem, because neoplastic transformation essentially involves replicating stem cells in tissues of renewal and because in the human brain such cells are found mostly in the course of CNS development. A cytogenetic scheme is therefore needed to serve as a frame of reference for a classification of embryonal CNS tumors that will account for the different histological entities documented so far and for the range and the restrictions of their differentiating capabilities. Most embryonal CNS tumors can be fitted into such a scheme. The cerebral medulloepithelioma, the cerebral and cerebellar neuroblastomas, the primitive polar spongioblastoma, and the ependymoblastoma show characteristic morphological features and a correspondingly distinctive cellular differentiating potential. The differentiating capabilities of the cerebellar medulloblastoma, the pineoblastoma, and the retinoblastoma are also distinctive, and are apparently determined by the cytogenesis of the area of the CNS in which the tumors originate. The indiscriminate application of a simplistic concept that would include all the so-called "primitive neuroectodermal tumors" into a single neuroepithelial tumor entity is unlikely to bring further understanding to the problem. PMID- 2987440 TI - Prediction of survival in glioma patients by means of positron emission tomography. AB - The aim of this study was to determine whether positron emission tomography (PET) with fluorine-18 (18F)-2-deoxyglucose (FDG) can be used as a prognostic test in patients with high-grade cerebral gliomas, regardless of the treatment given. Forty-five patients with astrocytoma Grade III or IV were included in this analysis. The mean survival time of patients with tumors exhibiting high glucose utilization as determined by PET-FDG was 5 months, whereas patients with gliomas showing lower glucose utilization had a mean survival period of 19 months. It is postulated that PET-FDG scans reflect the biological behavior of high-grade astrocytomas and may be used to predict the survival time of patients harboring such neoplasms. PMID- 2987441 TI - Variation in response to CCNU of glioblastoma multiforme in brain and cervical lymph node. Case report. AB - The case is reported of a patient in whom a cervical lymph node metastasis decreased in size while the primary intracranial glioblastoma continued to grow during chemotherapy with CCNU (1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea). This is the first such case reported in humans. Possible explanations for this phenomenon are discussed. PMID- 2987442 TI - Assessment of skin ulcer healing capability by technetium-99m phosphate angiogram and blood-pool images. AB - The accuracy of radionuclide angiography and blood-pool imaging using [99mTc] phosphate to assess skin ulcer perfusion as an indicator of healing capacity was determined in 50 studies performed on 45 patients with nonhealing ulcers of the lower extremities. Two nuclear medicine physicians without clinical bias, graded the perfusion of the ulcer on the images as normal, increased or reduced with respect to the opposite limb. Patients were followed closely with aggressive local wound care for at least 14 days. Of the 31 ulcers which healed, the radionuclide study correctly predicted 30; of the 19 ulcers which did not heal, 14 were correctly predicted. Eight patients had osteomyelitis; four of those healed and four did not. The radionuclide study predicted healing in seven. Excluding those patients with osteomyelitis, the sensitivity for the radionuclide angiogram and blood-pool image in predicting healing was 96%, specificity was 87%, and accuracy was 93%. This technique is a simple, reliable way to predict the microcirculatory adequacy for ulcer healing. Specificity is diminished in the presence of osteomyelitis. PMID- 2987443 TI - Research and clinical potential of receptor based radiopharmaceuticals. AB - Receptors are proteins that have specific binding affinity for substances that produce a physiological event in the body. Receptor-binding radiotracers are being used increasingly to study the function of receptors in health and disease. This review summarizes the proceedings of a symposium on research in the development of receptor-binding radiopharmaceuticals. The key phases in this research include: selection of the receptor system and ligand; synthesis of radiolabeled ligand; validation in animal models; and clinical application. Current research involves a variety of biological systems, such as butyrophenone neuroleptics for dopamine receptors and steroidal estrogens for the estrogen receptor. In the future, it is believed that receptor-binding radiopharmaceuticals will be useful, not only to validate receptor systems in vivo, but also to aid in the diagnosis and therapy of human diseases. PMID- 2987444 TI - Different protocols for parathyroid imaging. PMID- 2987445 TI - A learner analysis experiment: cognitive style versus learning style in undergraduate nursing education. AB - The purpose of this study was to determine whether or not statistically significant differences existed among performance levels of beginning undergraduate nursing students when compared according to cognitive style and learning style subgroups. The sample consisted of 60 nursing students enrolled in an introductory clinical nursing course at a regional community college in the Southwest. Subjects were randomly assigned to experimental and control groups and were administered separate learner analysis instruments. The 30 students in the control group were given a cognitive style mapping assessment, while the 30 students in the experimental group were given a learning style assessment. The control group and experimental group were both given group interpretations of the learner analysis data. As with all nursing students at the College, the control group was allowed individual learning prescriptions by the instructor. The experimental group was allowed no further data interpretation. Both groups were to use the learner analysis data to assist them in completing the course. Performance data were collected during the ensuing semester from five evaluations covering specified modules: "Basic Human Needs;" "Professional Ethics and Communications;" "Emergency Care, Fluids, and Electrolytes;" "Death and Oncology;" and "Pharmacology." Analysis of the data with t-tests revealed no significant differences (p = .05) in performance among the experimental or control groups. Mean scores from the evaluation data did indicate a slightly higher performance level from the control group in four of the five modules. PMID- 2987446 TI - Follow the leader: a learning exercise. AB - This manuscript presents an alternate method for students' acquisition of leadership concepts. It also invites the students to become actively invoLved in their own learning. "Follow the Leader: A Learning Exercise" enables students to observe leadership in action, and to analyze and assimilate leadership/management theories. After observing leaders, the students discuss how they would use what they learned to become better nursing leaders. The exercise covers these key nursing management concepts: principles of delegation of authority and responsibility, communication patterns, group process, decision making, and evaluation. In order to do the exercise, students need background knowledge of basic management procedures of planning, organizing, directing and controlling. PMID- 2987448 TI - Crisis intervention: when the client is a nursing student. PMID- 2987447 TI - Defining scholarly activity in nursing education. AB - In an attempt to address the question of what activities should be considered scholarly, thereby warranting their inclusion in a nursing faculty evaluation model, a study was undertaken which surveyed all National League for Nursing accredited baccalaureate educational programs. The items in the instrument were generated utilizing the Delphi method and a pilot study established inter respondent reliability. A response rate of 73% was attained. Scholarly activity was considered highly important in evaluation for promotion and tenure in over 50% of the schools. There were distinct differences in the activities deemed scholarly when respondents were broken down into categories such as size and type of institution and the existence or non-existence of graduate nursing programs in the same institution. PMID- 2987449 TI - Toward more clarity in terminology: frameworks, theories and models. AB - Lack of precision in a person's choice of words, which perhaps indicates a lack of clarity and accuracy in ideas, may become a subtle form of negative teaching. It is suggested that such terms as "conceptual frame of reference" and "theoretical framework" should be used as broad terms for conceptions of reality, reserving "philosophy," "theory" and "conceptual model" as more restricted terms for conceptions of reality. Nurse authors do not agree that nursing theory exists, that nursing has its own body of knowledge, that nursing is a science or even a discipline. Yet nursing is fully equipped to justify its claim to such terms. Starting from a nursing perspective rather than from a borrowed one, nursing theory can be developed, nursing knowledge can be accumulated, and nursing science can be advanced, all in the interests of a worthwhile contribution to society's health. PMID- 2987450 TI - Guiding students in theory development through conceptual mapping. PMID- 2987451 TI - Helping the pregnant student stay in school. PMID- 2987452 TI - Simulation in teaching clinical nursing. AB - Clinical simulation research is active in nursing. This research has yielded certain curricular implications. Simulated experiences are very effective in identifying student strengths and weaknesses, as well as curricular strengths and weaknesses. In addition, simulated nursing experiences provide learning in a controlled environment that increases the student's confidence and enhances patient safety and comfort. In this paper, the simulated test has been discussed in the context of a generic nursing student; however, there are definite possibilities for validating the knowledge of registered nurse students, new graduates in specialty areas, and expert nurses taking certification exams. In conclusion, the author views simulation (whether written, on computer, as a game, or practiced on mannequins) as a means of bringing clinically-related experiences into the school, and as being a more efficient and safer method of providing clinical experiences. It is apparent that many schools of nursing are utilizing simulation teaching and evaluation as a complement to patient contact experiences. Simulation is being used with a variety of subjects and concepts and with a wide range of student abilities and classifications. It has proven to be efficient and effective in these days of tight time schedules, "full" curricula, and crowded clinical facilities. PMID- 2987454 TI - Interactive clinical conferences: nursing rounds and education imagery. PMID- 2987453 TI - Teaching communication skills in an integrated curriculum. AB - During the seven weeks students showed growth in empathic responding. In addition, many students were able to move beyond beginning empathy to identifying underlying feelings experienced by the patient. Progress in concreteness was noted as irrelevant questioning decreased and selective, open-ended questioning increased. There seemed to be a positive correlation between self-awareness and growth in communication skills. Initially, use of behaviorally specific feedback helped the students identify strengths and weaknesses. As the sessions progressed, students became more independent in critiquing their own interpersonal style. For example, one student identified her abrasive manner in a taped interaction and recognized her need to maintain control. She modified her approach and became more caring and empathic. Initially, Stage II skills were difficult for the students. Some students who expected to see change occur quickly, became impatient with relationship building, and used challenging skills prematurely. Other students had difficulty taking the risk to challenge the patient to focus on problem exploration. Role playing in the training sessions and faculty feedback in the clinical settings helped the students begin to use skills more appropriately. Observing the positive effects on patient behavior reinforced continued use. Students and faculty were enthusiastic about the workshop format and outcome. Student reactions reinforced the value of the systematic approach. Through verbal and written evaluations they identified the small group practice and clinical audiotaping as the most valuable activities. Many liked being able to practice and receive immediate feedback from their peers and instructors. Faculty found that the four components-description, demonstration, practice and application-provided direction for teaching.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987455 TI - Interventions useful to the public health nurse: improving health behaviors. PMID- 2987456 TI - Modification of a simulation game for use in a large group setting. PMID- 2987457 TI - Perceptions of a geriatric rotation: influence on career choices. PMID- 2987458 TI - Values clarification: a critique. PMID- 2987459 TI - Composition of fecal fiber from human subjects. AB - This research addresses two problems of fecal fiber analysis: the nitrogen in fecal neutral detergent fiber (NDF); and the impact of sodium sulfite on NDF yield and composition. Fecal samples for analysis were collected from healthy adults during a study of the effect of a wheat bran supplement on bowel function. When sodium sulfite, originally a component of neutral detergent solution, was omitted, the yield of fecal NDF, the concentration of apparent lignin in the NDF and the amount of nitrogen recovered in both the NDF residue and the lignin increased. These data support a previous report by Van Soest that the sulfite attacks lignin. We present evidence that the effect of sodium sulfite also could be one of inhibiting the formation of Maillard products, which are detected as lignin, during the reflux step of the NDF method. Three different fractions of nitrogen were identified in fecal NDF: the fraction eliminated by sodium sulfite; a fraction removed by dilute acid; and the fraction that remained in Klason lignin. Apparent NDF digestibilities were calculated several ways by using correction factors to account for this nitrogen. Only the subtraction of Klason lignin from fecal NDF eliminated the one negative digestibility we observed. PMID- 2987460 TI - TORCH: a literature review and implications for practice. AB - Major issues related to TORCH (toxoplasmosis, rubella, cytomegalo virus, and herpes virus) perinatal infection are of concern to maternity and neonatal nurses. General and congenital modes of transmission, associated congenital defects, incidence of infection, and relative fetal/neonatal effects of primary as compared to recurrent maternal infection are described. Associated nursing implications emphasizing prenatal prevention and early detection, intrapartal management, neonatal symptom detection, and nursery infection control as well as concerns about breastfeeding with maternal TORCH are discussed. PMID- 2987461 TI - ABC's periodontics: "K" is for the "Keyes Technique". PMID- 2987462 TI - A technique for controlled placement of hydroxylapatite over atrophic mandibular ridges. AB - When hydroxylapatite is injected into a subperiosteal tunnel without placing buccolingual sutures, there is occasionally noticeable drifting of the material outside the desired area, and the labial vestibule may be diminished. By placing horizontal mattress sutures, the hydroxylapatite is controlled and ridge contour is improved. There appears to be no added morbidity from placement of the buccolingual sutures. However, because only a small number of cases have been treated with this procedure, this report should be viewed as preliminary in nature. PMID- 2987463 TI - Effect of inflammation upon human gingival cyclic AMP metabolism. AB - The effect of the increasing degree of human gingival inflammation on adenylate cyclase (basal, fluoride stimulated) and low Km and high Km cAMP phosphodiesterase activities were evaluated in separate studies. Human gingival biopsies were classified by the Loe Bleeding Index as mildly, moderately, and markedly inflamed. Basal and F- stimulated adenylate cyclase (cAMP synthesis) activities were found to be unaltered by the increasing degree of inflammation when the data were expressed on either a mg wet wt, or mg protein basis. A significant loss of F- stimulated adenylate cyclase (mg protein) activity was observed in the moderately inflamed group when the data were compared with either the mildly or markedly inflamed groups of tissue. The low Km, and high Km cAMP phosphodiesterase activities (cAMP degradation) were found to be unaffected by gingival inflammation. This suggests that neither cAMP synthesis, nor degradation are stimulated in human gingiva by inflammation. PMID- 2987464 TI - Matrix vesicle enzymes in human osteoarthritis. AB - The enzymatic activities and in vitro calcification properties of matrix vesicle fractions isolated from normal and osteoarthritic (OA) human articular cartilage were compared to determine the essential conditions for calcification in these tissues. Four groups of human cartilage were examined, I, normal articular cartilage from aged, nonOA joints; II, discolored or fibrillated cartilage from OA joints; III, osteophytic cartilage from OA joints; IV, loose body cartilage from OA joints. Fetal bovine growth plate cartilage was also studied. Both ATP- and 5'-AMP-dependent in vitro matrix vesicle calcification occurs in all cartilage groups examined and, for human articular cartilage, these activities increase progressively from Groups I to II to III. Calcification does not occur in the absence of either phosphate or pyrophosphate. Alkaline phosphatase, 5' AMPase, and ATP:pyrophosphohydrolase activities are increased in Groups III and IV cartilage compared with Group I and are detected at high levels in fetal bovine growth plate cartilage. Pyrophosphatase activity occurs in only those cartilage groups juxtaposed to areas of new bone formation (osteophytic, loose body, and bovine growth plate). These results suggest that OA, growth plate, and even normal articular cartilage all have the potential to undergo calcification as long as both phosphate and pyrophosphate ions can be generated at sufficiently high levels. However, the capacity for cartilage to deposit hydroxyapatite, as it does during bone formation, may depend on the presence of pyrophosphatase activity. PMID- 2987466 TI - [Transmission of Epstein-Barr virus specific antigen associated with cell fusion activity]. PMID- 2987465 TI - Degradative enzyme systems in osteoarthritic cartilage. AB - In osteoarthritis, despite increased matrix synthesis, there is a reduction of both major matrix components, proteoglycan and collagen. This study suggests that this is the result of enhanced degradative activity intrinsic to the cartilage. Because osteoarthritis is a focal disease, histologic controls were used to measure the severity in different areas of the cartilage and in different specimens. Neutral proteoglycanase and collagenase were both described in human cartilage, and their levels matched the severity of the disease as did acid phosphatase, a marker of lysosomal enzymes. Articular cartilage collagenase has an inhibitor in the cartilage and has negligible activity in normal cartilage. This was found not to be a lysosomal enzyme. A model of osteoarthritis was studied and found to have the same biochemical pattern as human disease. Using this method, inhibitors of degradative enzymes were used as a treatment. The chelator of metallic cations EDTA was found to have a significant effect on reduction of degradative enzyme activity and altered the arthritic process. PMID- 2987467 TI - [Measurement of beta-adrenoceptors in lymphocytes of subjects with nasal allergy]. PMID- 2987468 TI - Usefulness of basement membrane markers in tumoural pathology. AB - The distribution of basement membrane (BM) markers, type IV collagen, laminin (LM), heparan sulphate proteoglycan (HSP) and fibronectin (FN) has been studied by indirect immunofluorescence using specific antibodies, in tumoural pathology. The disrupted pattern of BM by these markers in severe dysplastic lesions of the breasts, the bronchi and uterine cervix provides evidence for malignancy. In invasive carcinomas, there is generally a loss of these BM components, with FN persisting in the stroma. The loss of these markers in BM is concomitant and superimposable in double staining studies. In embryonic tumours, the presence of BM markers is related to a mesenchymal differentiation of malignant cells with pericellular FN and/or maturation towards organoid structures with BM. In sarcomas, there is a loss of the pericellular BM staining around most transformed muscular and Schwann cells and adipocytes. The persistence of this labelling in some well-differentiated areas can help to diagnose the nature of the sarcoma. The persistence of intercellular filaments of FN corresponds to the mesenchymal and/or sarcomatous nature of undifferentiated anaplastic proliferations. PMID- 2987469 TI - The effects of calcium regulating hormones on bone resorption by isolated human osteoclastoma cells. AB - Cells were disaggregated from osteoclastomas, and the response of the giant cells to calcium-regulating hormones, prostaglandin (PG)E1 and dibutyryl cyclic AMP (dbcAMP) was observed by phase-contrast time-lapse video microscopy. The pattern and nature of their response was very similar to that previously found to be characteristic of osteoclasts: calcitonin (CT), PGE1 and dbcAMP induced cytoplasmic quiescence, while parathyroid hormone (PTH) showed no influence on cytoplasmic motility or behaviour. The cells were also cultured on slices of devitalized cortical bone for 5 or 18 h. After this time the giant cells were associated with the appearance in the scanning electron microscope of characteristic resorption pits, the volume of which was calculated by computer assisted morphometric and stereophotogrammetric techniques after removal of cells. Calcitonin caused a dramatic reduction in the volume of bone resorbed by these isolated cells compared with control cultures, while PTH was without significant effect. This result supports the view that PTH does not increase bone resorption in intact bone through a direct effect on osteoclasts. PGE1, which stimulates bone resorption when added to intact bone, paradoxically reduced resorption in our cultures. It thus appears possible that PGE1 acts as a direct inhibitor of osteoclastic bone resorption but has an additional effect on other cells in bone, which are induced by PGE1 to cause osteoclastic stimulation. PMID- 2987471 TI - Hepatocellular carcinoma in a noncirrhotic infant after prolonged parenteral nutrition. PMID- 2987472 TI - Generalized odontodysplasia. PMID- 2987473 TI - Improved technique for localized ridge augmentation. A report of 21 cases. AB - Topographical aberrations in a residual edentulous ridge often prevent establishment of a satisfactory pontic/ridge relationship. An improved technique is described for predictable augmentation of localized alveolar-ridge deficiencies. Results are reported from 21 cases involving 26 sites. All 14 sites using fibrous connective tissue grafts demonstrated shrinkage, although an improvement in residual ridge contour was obtained. Hydroxylapatite implant material was placed in 12 sites with shrinkage seen in only two sites. Advantages, requirements for success and technical considerations of the improved technique are discussed. PMID- 2987470 TI - Atypical hypocomplementemic vasculitis syndrome in a child. AB - We report a patient who developed recurrent urticaria and angioedema at age 2 years, severe hypocomplementemic glomerulonephritis at 11 years, and end-stage renal disease at 14 years. His disease resembled the hypocomplementemic vasculitis syndrome but was atypical in its early age of presentation, severe hypocomplementemia, and progression to end-stage renal disease. Serum C1q levels were extremely low, and C4, C2, C3, and C5 levels were significantly reduced. Serum C1 inhibitor (C1INH) levels were slightly low, presumably from consumption. Circulating C1INH-C1r-C1s complexes were evidenced by reduced ratios of functional to antigenic C1INH and antigenic C1r to C1s. Family members had normal functional and antigenic levels of all complement components studied. The patient's serum, erythrocytes, platelets, and mononuclear cells did not activate complement when mixed with normal target serum. Absence of a circulating complement activator and the low serum C3 and C5 levels suggested the presence of a solid-phase complement activator, possibly related to renal or systemic vascular endothelium. As in patients with homozygous deficiencies of classical pathway components, a severe, prolonged, acquired C1q deficiency may have predisposed this patient to the development of glomerulonephritis. PMID- 2987474 TI - Repair of anterior gingival deformity with durapatite. A case report. AB - After the extraction of a maxillary anterior tooth, a gingival deformity may occur due to the loss of the labial plate of bone. This often creates an esthetic problem in the construction of a fixed partial denture. In the past, various surgical techniques have been devised to eliminate this gingival defect. A technique using durapatite is discussed. PMID- 2987475 TI - Alpha adrenoceptor number limits response of some rabbit arteries to norepinephrine. AB - Some properties of the receptor population mediating contraction to I norepinephrine (NE) were studied in rabbit aorta and the intrapulmonary and basilar arteries. The constrictor effect of NE in isolated vessel ring segments were recorded, both in the absence and presence of concentrations of phenoxybenzamine which depressed the maximal contractions to NE. Dose-response curves to NE were biphasic in the basilar artery. In the basilar artery and in medium-sized intrapulmonary arteries, the response to maximally effective concentrations of NE were always lower than responses to other agents. In the larger intrapulmonary artery segments and aorta the responses to NE and serotonin were of similar magnitude. The ED50 for NE did not change with vessel size in intrapulmonary arteries, whereas that of the basilar artery was considerably greater than in the pulmonary and aorta segments. The NE dissociation constant (KA) and the receptor occupation-response relationship was calculated in all vessel segments. The KA for NE was similar in part I of the basilar artery, aorta and both sizes of pulmonary artery. The KA for NE in part II of the basilar artery dose-response curves was 8 to 10 times greater. Half-maximal responses by NE were produced by occupation of 10% of receptors in the aorta and the large intrapulmonary artery and of 40 to 50% receptors in the basilar and medium-sized intrapulmonary artery. It is concluded that in some smaller arteries the small NE response may be limited by receptor number, and that whereas this may not be a phenomenon common to all vessels of this size, it may reflect specialization of certain vascular beds. PMID- 2987476 TI - Acute and subchronic effects of MJ-13859, a potential antipsychotic drug, on rat brain dopaminergic function. AB - The potential antipsychotic drug, MJ-13859, was tested for its acute and subchronic effects on rat brain dopaminergic neurotransmission and function. This drug exhibited an IC50 for displacement of [3H]spiperone binding of 6.35 nM which was similar to trifluoperazine, a potent classical antipsychotic drug. In female rats, MJ-13859 was slightly less potent than trifluoperazine for induction of catalepsy, inhibition of stimulant-induced hyperactivity and for increasing striatal, frontal cortical and olfactory tubercule dopamine metabolism. Both drugs also blocked dopamine autoreceptors on striatal dopamine nerve endings. When administered i.p., rather than s.c., the effect of MJ-13859 on dopamine metabolism was reduced significantly. When administered to male rats, the response of dopamine metabolism to drug was reduced, but a similar inhibition of stimulated activity occurred as in female rats. These results suggest a rapid first pass metabolism of MJ-13859 in the rat and that the antistimulant effect may be partly independent of the antidopaminergic effects. Osmotic minipumps were implanted s.c. for 2 week continuous infusion of MJ-13859 at doses of 1.0 or 3.0 mg/kg/day. Unlike classical antipsychotic drugs, MJ-13859 did not cause a subsensitivity to the ability of acute haloperidol challenge (0.1 mg/kg s.c.) to increase dopamine metabolism. After allowing a 4-day drug washout period before [3H]spiperone binding assay for D2 receptors, neither the maximum binding nor Kd were altered in rats treated for 2 weeks with 3.0 mg/kg/day of MJ-13859. Haloperidol at 1.0 mg/kg/day for 2 weeks caused a 57% increase in D2 receptor maximum binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987477 TI - Reduction of the cyclic adenosine 3',5'-monophosphate response to catecholamines in rat brain slices after repeated restraint stress. AB - Restraint stress reduces the cyclic AMP (cAMP) response to norepinephrine (NE) in slices of the rat cerebral cortex and hypothalamus. This effect is found after repeated but not single exposure to stress and persists for at least 24 hr poststress. The magnitude of the reduction is dose-dependent in that greater decreases are found after higher frequencies and longer durations of restraint as well as after more disturbance during the stress. Analysis of the NE-cAMP dose response curve indicates that the stress reduces the maximum cAMP response to NE but does not increase the EC50 value of NE. The cAMP response to isoproterenol is only slightly affected by the stress. No effect is observed on specific [3H]dihydroalprenolol binding in either brain region at 24 hr poststress. These results suggest that repeated restraint stress produces a selective persistent reduction of the function of brain non-beta adrenergic receptors. This effect may be mediated by an increased release of adrenocorticotropic hormone as chronic infusion of the latter hormone mimics the action of stress on cAMP responses to catecholamines. An increased release of brain NE may also be involved as repeated administration of the NE-reuptake inhibiting antidepressant, desmethylimipamine, reduces the function of non-beta as well as beta adrenergic receptors as evidenced by reductions of both the NE- and isoproterenol-cAMP responses. PMID- 2987478 TI - Selectivity of (+)-4-propyl-9-hydroxynaphthoxazine [(+)-PHNO] for dopamine receptors in vitro and in vivo. AB - The intrinsic activity of the potent dopamine (DA) agonist 4-propyl-9 hydroxynaphthoxazine [(+)-PHNO] was examined in receptor binding assays for the following receptors: DA, alpha-1 and alpha-2 adrenergic, serotonin-1 and -2, neuroleptic, beta adrenergic, anxiolytic, adenosine A-1, gamma-aminobutyric acid, muscarinic and opiate. (+)-PHNO exhibited strong to moderate potencies [IC50 (nanomolars) in parentheses] in binding to DA (24), "neuroleptic" (67), alpha-2 adrenergic (77) and serotonin-1 (277) sites. The pharmacological activity of the naphthoxazine both in vivo and in vitro was contrasted with the known DA agonists apomorphine, pergolide, lisuride and 6-ethyl-9-oxaergoline in tests of DA, alpha 2 adrenergic and serotonergic function. Each compound was examined in vitro in receptor binding assays for interactions with DA, alpha-2 adrenergic, serotonin-1 and serotonin-2 receptors and for alpha-2 adrenergic activity in inhibiting field stimulated contractions of the vas deferens of the rat. In vivo, alpha-2 adrenergic activity was assayed via measurement of mydriasis after i.v. injections in the rat, whereas serotonin activity was assayed by measuring drug induced inhibition of 5-hydroxytryptophan accumulation, and DA activity was assessed by quantifying stereotyped behavior after both i.p. and i.v. injections. Selectivity ratios for the DA receptor were derived from effective dose values determined in these tests and demonstrated that only apomorphine was more selective as a DA agonist than (+)-PHNO in vivo. (+)-PHNO was the least active agent at the alpha-2 receptors in the vas deferens and with serotonergic mechanisms in vivo to reduce 5-hydroxytryptophan accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987479 TI - Neuronal survival during electrical blockade is increased by 8-bromo cyclic adenosine 3',5' monophosphate. AB - Dissociated cultures from spinal cord-dorsal root ganglion were used as a model system to study the interaction between electrical activity and cyclic nucleotides on neuronal survival. Neuronal death produced by electrical blockade with tetrodotoxin (TTX) was attenuated by the addition of 8-Bromo cyclic AMP (8 Br cAMP). [125I]Tetanus toxin fixation, choline acetyltransferase activity and neuronal cell counts were used to quantitate the effects of 8-Br cAMP on the cultures. A dose-dependent increase in both tetanus toxin fixation (neuronal surface marker) and choline acetyltransferase activity was observed with 8-Br cAMP application to TTX-treated cultures as compared to those treated with TTX alone. The rate of choline acetyltransferase induction by 8-Br cAMP was decreased in the presence of TTX. Supplementation of the cultures with 8-bromo cyclic GMP, under the same conditions, had no protective or inductive effects. These data suggest that cholinergic neurons are among those cells affected by electrical blockade and by increased culture cAMP levels. Electrical blockade with TTX produced significant decreases in cAMP levels during a critical period in development. The onset of the critical period was found to be between day 5 and day 8 in vitro and the vulnerability to TTX-mediated decreases in cAMP were not evident in cultures which were older than 1 month. Conditioned medium obtained from spinal cord-dorsal root ganglion cultures before the critical period were tested on TTX-treated cultures that were within the critical period. No decrease in cAMP levels from that of controls was observed after 50% conditioned medium plus TTX treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987480 TI - Inhibitory effect of methacholine on drug-induced relaxation, cyclic AMP accumulation, and cyclic AMP-dependent protein kinase activation in canine tracheal smooth muscle. AB - Functional antagonism between bronchoconstricting and bronchodilating pathways was examined in canine tracheal smooth muscle. Trachealis strips were contracted with either 0.3 microM (EC55) or 3.0 microM (EC80) methacholine before being relaxed by the cumulative addition of isoproterenol, prostaglandin E2, or forskolin. The EC50 for all three relaxants was increased 10-fold in tissues contracted with 3.0 microM methacholine vs. those contracted with 0.3 microM methacholine. Moreover, contracting tissues with the higher concentration of methacholine reduced the maximum relaxation induced by prostaglandin E2 and isoproterenol. Forskolin produced total relaxation regardless of the concentration of methacholine used and thus was a much more effective bronchodilator than either isoproterenol or prostaglandin E2. The inhibitory effect of methacholine on the relaxant response to these agents was paralleled by a reduction in drug-stimulated cyclic AMP-dependent protein kinase activity. Methacholine reduced the maximum activation of cyclic AMP-dependent protein kinase elicited by isoproterenol, prostaglandin E2 and submaximal concentrations of forskolin, which was a much more powerful enzyme activator than the other two agents. The ability of a maximum concentration of forskolin (30 microM) to activate cyclic AMP-dependent protein kinase was not inhibited by methacholine. Although methacholine also appeared to suppress drug-stimulated cyclic AMP accumulation, the inhibitory effect was only statistically significant in forskolin-treated tissues.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987481 TI - Pharmacologic characterization of human and canine thromboxane A2/prostaglandin H2 receptors in platelets and blood vessels: evidence for different receptors. AB - The present study was undertaken to characterize thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptors in platelets and blood vessels. Both human and canine platelet aggregation and saphenous vein contractions were induced by the stable TXA2/PGH2 mimetics (15S)-hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5Z, 13E dienoic acid (U46619) and 9,11-epithio-11,12-methano-TXA2 (ONO-11113). ONO-11113 was a more potent agonist than U46619 in the human saphenous vein but less potent in the platelet. These agonists were equipotent in the canine platelet but ONO 11113 was more potent in the saphenous vein. Platelet aggregation and saphenous vein contraction induced by U46619 were blocked in a dose-dependent manner by the TXA2/PGH2 receptor agonists 9,11-dimethylmethano-11,12-methano-16-phenyl-13,14 dehydro-13-aza- 15 alpha beta-omega-tetranor-TXA2 (ONO-11120), 9,11 dimethylmethano-11,12-methano-16-(4-hydroxyphenyl)-13,14-d ihy dro-13-aza-15 alpha beta-omega-tetranor-TXA2 (PTA-OH), 9,11-dimethylmethano-11,12-methano-16-(4 methoxyphenyl)-13,14-d ihy dro-13-aza-15 alpha beta-omega-tetranor-TXA2 (PTA-OM), 9,11-dimethylmethano-11,12-methano-16-(3-iodo-4-hydroxyphenyl-13,1 4-dihydro-13- aza-15 alpha beta-omega-tetranor-TXA2 (I-PTA-OH) and 9,11-dimethylmethano-11,12 methano-15-phenyl-13,14-dihydro-13-aza- 15 alpha beta-omega-pentanor-TXA2 [PTA (omega-1)].(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987482 TI - In vivo binding of benzomorphans to mu, delta and kappa opioid receptors: comparison with urine output in the rat. AB - In vivo binding affinities of three benzomorphans, Win 44,441-3, bremazocine and MR 2266, were determined at the mu, delta and kappa types of opioid binding sites in rat brain, using an ex vivo labeling technique. The receptor occupancy of the benzomorphans and of previously tested diprenorphine were compared with their activities in increasing urine output (agonist ED50: bremazocine) or inhibiting bremazocine-induced diuresis (antagonist ID50: Win 44,441-3, MR 2266 and diprenorphine). The agonist, bremazocine, bound (in order of decreasing affinity) to the kappa approximately equal to mu greater than delta binding sites, and it's pharmacological effects appeared in the dosage range of kappa and mu binding. In order to positively identify which receptor type is responsible, the potency of the three antagonists to block the effects of bremazocine were compared to their ability to occupy the individual sites in vivo. A fractional occupancy of 0.5 would be expected at the ID50 if one assumes a linear relationship between receptor occupancy by the antagonist and the antagonistic effect. Such a linear relationship was observed for the three antagonists only at the kappa site, whereas variable occupancies were observed at the mu and delta sites. These results support the previously proposed hypothesis that kappa receptors mediate the effects of benzomorphan opioid drugs on urine flow. PMID- 2987483 TI - Lack of desensitization of alpha-2-mediated inhibition of lipolysis in fat cells after acute and chronic treatment with clonidine. AB - The regulation of alpha-2 adrenoceptors and alpha-2 adrenergic responses by alpha 2 agonist treatment was investigated in in vitro and in vivo conditions. The alpha-2 adrenergic responsiveness of the adipocytes was tested with an alpha-2 agonist, clonidine, which inhibits, in a concentration-dependent manner, the lipolytic activity of the isolated fat cells incubated in the presence of adenosine deaminase (1-2 micrograms/ml). The effect of in vitro treatment of human s.c. fat cells and hamster adipocytes with clonidine as well as the incidence of chronic clonidine treatment (10 days) of golden hamsters were studied by testing lipolytic and antilipolytic responses of the isolated fat cells. Moreover, binding parameters were determined on the corresponding fat cell ghosts using [3H]clonidine and [3H]yohimbine. The preincubation of hamster (90 min) and human (3 hr) fat cells with clonidine did not modify the biological responses promoted by isoproterenol or clonidine. Binding analysis showed that the number of [3H]clonidine sites was reduced by 30 to 40% in human fat cell ghosts whereas [3H]yohimbine sites were unaffected by clonidine treatment. The chronic treatment of hamsters with clonidine (10 days, 200 micrograms/day) did not influence the lipolytic and antilipolytic responses of the fat cells. However, the number of alpha-2 adrenoceptor sites, identified by [3H]clonidine was reduced (40-50%). To conclude, short-term incubations of adipocytes with clonidine or long-term in vivo treatments with this drug are without any noticeable influence on the lipolytic and antilipolytic responses of human and hamster isolated fat cells. The reduction of [3H]clonidine binding sites described in both experiments must be questioned concerning its biological relevance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987484 TI - Demonstration of an alpha adrenoceptor-mediated inotropic effect of norepinephrine in human atria. AB - It has been claimed by other investigators that norepinephrine does not evoke a significant alpha adrenergic inotropic effect in human atria in contrast to epinephrine and phenylephrine, indicating a limitation of a possible functional role of the cardiac alpha adrenoceptors. We therefore characterized the inotropic effects of norepinephrine in isometrically contracting muscle strips from human atria obtained during open heart surgery. Both contraction and relaxation were studied by measuring developed tension and its first and second derivatives. Both the influence of propranolol and prazosin upon the inotropic responses to norepinephrine and the qualitative characteristics of the responses revealed that norepinephrine evoked both alpha and beta adrenergic inotropic effects. The alpha adrenergic response to norepinephrine was qualitatively different from the beta adrenergic effect and qualitatively similar to the alpha adrenergic effect of norepinephrine observed in other mammalian species. Although the alpha adrenergic effect was marked, the beta adrenergic effect was the dominating one as has also been found in other species. It is concluded that also in human atria norepinephrine evokes inotropic effects through both alpha and beta adrenoceptors. PMID- 2987485 TI - Inhibition by 5,6-dihydroxy-2-dimethylaminotetralin (M7) of noradrenergic neurotransmission in the rabbit hypothalamus: role of alpha-2 adrenoceptors and of dopamine receptors. AB - Rabbit hypothalamic slices were prelabeled with [3H]norepinephrine and transmitter release elicited by electrical stimulation. In the presence of 10 microM cocaine and in a low Ca++ medium (0.65 mM), exposure for 8 min to exogenous dopamine (0.01-1 microM) inhibited, in a concentration-dependent manner, the electrically evoked release of [3H]norepinephrine. This inhibitory effect of dopamine on [3H]norepinephrine release was antagonized by the dopamine receptor antagonist S-sulpiride (1 microM), but remained unchanged in the presence of the alpha-2 adrenoceptor antagonists idazoxan (1 microM) or yohimbine (0.1 microM). These results indicate that, in a low Ca++ medium, exposure to dopamine decreased [3H]norepinephrine overflow in rabbit hypothalamic slices through the exclusive activation of presynaptic inhibitory dopamine receptors. M7 (5,6-dihydroxy-2-dimethylaminotetralin) is a potent agonist at central presynaptic dopamine autoreceptors and at peripheral alpha-2 adrenoceptors. Exposure to M7 in a normal Ca++ medium, inhibited in a concentration-dependent manner the electrically evoked release of [3H]norepinephrine without affecting the spontaneous outflow of radioactivity. The slope of the concentration-effect curve for these inhibitory effects of M7 was rather flat and the maximal inhibition obtained was 80%. The selective D2 receptor antagonist S-sulpiride (1 microM) failed to produce a significant shift to the right in the concentration effect curve for the inhibitory effects of M7 on [3H]norepinephrine release. The preferential alpha-2 adrenoceptor antagonist yohimbine (0.1 microM) significantly antagonized the inhibition of [3H]norepinephrine release elicited by 0.01 microM M7, but not for higher concentrations of this aminotetraline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987486 TI - Chronic thyroxine treatment of rats down-regulates the noradrenergic cyclic AMP generating system in cerebral cortex. AB - Treatment of Sprague-Dawley rats for 9 days with 15 micrograms/kg of tri-iodo thyronine, 50 micrograms/kg of thyroxine (T4) or 500 micrograms/kg of thyrotropin releasing hormone decreased the number of beta receptors in cerebral cortex as measured by dihydroalprenolol binding. The dissociation constants of dihydroalprenolol (3.5 nM) were not altered by the treatment. Only tri-iodo thyronine and T4 administration resulted in a concomitant reduction of norepinephrine (NE) elicited cyclic AMP formation in cerebral cortical slices. This process required at least a 7-day treatment period and was dose-dependent. Application of 1 microgram of T4 per kg for 9 days significantly diminished the NE responsivity of the cyclic AMP synthesizing system. Dose-response curves with NE indicate a reduction of the maximal response after T4 treatment with no change in ED50. An almost additive interaction between the effects of T4 and a low dose (3 mg/kg) of the tricyclic antidepressant imipramine was observed. Striking differences in the response of the adrenoceptor coupled adenylate cyclase to a 9 day T4 treatment were found when different rat strains, i.e., Fischer F-344, Long Evans, Wistar and Sprague-Dawley were used. The hyperthyroid state of the animals was ascertained by measurements of plasma levels of tri-iodo-thyronine and T4. Down-regulation of NE sensitive adenylate cyclase by T4 treatment required intact synaptic structures because denervation by i.c.v. injection of 6-hydroxydopamine abolished the effect of T4 treatment. This is indicative of a postsynaptic localization of the down-regulated cyclic AMP generating system. The data stress the importance of the neuroendocrine system for adrenoceptor regulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987487 TI - 5-Hydroxytryptamine-stimulated inositol phospholipid hydrolysis in rat cerebral cortex slices: pharmacological characterization and effects of antidepressants. AB - The ability of 5-hydroxytryptamine (5-HT) and related agonists to stimulate hydrolysis of inositol phospholipids was examined in rat cerebral cortex slices using a direct assay which involves prelabeling with [3H]inositol and assaying [3H]inositol phosphates in the presence of lithium. 5-HT agonists stimulated [3H]inositol phosphate accumulation in a dose-related but biphasic manner and only the high-affinity component of the dose-response curve was sensitive to antagonists. This response to 5-HT was blocked potently by ketanserin and other putative 5-HT2 antagonists but the overall pattern of apparent drug affinities was inconsistent with that seen at 5-HT2 sites labeled with [3H]ketanserin in cortical membranes. Pretreatment of slices with the alkylating antagonist phenoxybenzamine reduced the inositol phospholipid response to 5-HT to a greater extent than the suppression of [3H]ketanserin binding. Similarly, chronic but not acute treatment of rats with the antidepressants iprindole and imipramine resulted in a greater loss of 5-HT-induced inositol phospholipid hydrolysis than specific [3H]ketanserin binding. However, the effect of antidepressants was agonist-specific in that neither alpha-1 adrenoceptor nor muscarinic receptor stimulation were altered by acute or chronic treatment. PMID- 2987488 TI - Peripheral-type benzodiazepine receptors: autoradiographic localization in whole body sections of neonatal rats. AB - We have developed a procedure that allows the autoradiographic localization of benzodiazepine receptors in whole-body sections of neonatal rats. Central-type benzodiazepine receptors, visualized with [3H]methylclonazepam, are restricted to nervous tissue. In contrast, peripheral-type benzodiazepine receptors, visualized with [3H]Ro5-4864, occur widely, but with discrete localizations throughout the body. Peripheral-type benzodiazepine receptors are most concentrated in the adrenal cortex and the skin. Substantial levels of these receptors are also evident in the heart, the salivary glands, discrete regions of the kidney, the epithelium of the lung, the nasal and lingual epithelia, the lining of the pulmonary arteries, the thymus, the hair follicles of the vibrissae, the tooth buds and the bone marrow. Considerable binding of [3H]Ro5-4864 is observed in the brown fat pads, the liver and the spleen, but high levels of nonspecific binding preclude accurate evaluation of the actual specific binding in these organs. Only low levels of [3H]Ro5-4864 binding sites are found in the brain and they are virtually undetectable in the skeletal muscle, the eye, the inner ear and the gastrointestinal tract. High levels of peripheral-type benzodiazepine receptor appear present in tissues that derive their metabolic energy primarily from oxidative phosphorylation, whereas only low levels are present in tissues that can derive their metabolic energy largely from glycogenolysis. Association of these receptors with mitochondria and a possible role in modulation of energy metabolism is suggested further by the observation that the histochemically visualized distribution of cytochrome oxidase activity overlaps the autoradiographic pattern of [3H]Ro5-4864 binding sites. PMID- 2987489 TI - Electrical properties of rat dorsal root ganglion neurones with different peripheral nerve conduction velocities. AB - The electrical characteristics of individual rat dorsal root ganglion neurones were studied and related to the peripheral axon conduction velocity and morphological cell type. Neurones were divided into four groups based on the conduction velocity of their peripheral axons (A alpha, 30-55 m/s; A beta, 14-30 m/s; A delta, 2.2-8 m/s and C less than 1.4 m/s). Electrophysiological parameters examined included membrane potential, action potential amplitude and duration, after-potential height and duration, input resistance and the occurrence of time dependent rectification. The mean duration of the somatic action potentials was found to be characteristic for each of the conduction velocity groupings. However, there was considerable overlap between groups. The fast-conducting (A alpha) and slowly conducting (A delta) myelinated fibres had short-duration action potentials, within the ranges 0.49-1.35 and 0.5-1.7 ms at the base respectively. The A beta and C cells had somatic action potentials with durations in the ranges of 0.6-2.9 and 0.6-7.4 ms respectively. The longer action potential durations could be related to the presence of an inflexion on the repolarizing phase seen in a third of A beta neurones (called A beta I neurones) and in all C neurones. The action potential overshoot was larger in C neurones and A beta I neurones than in the other neurone groups. The mean duration of the after hyperpolarization was several times greater in C neurones than in A neurones. A delta neurones displayed the shortest and greatest amplitude after hyperpolarizations. Large, long-lasting after-hyperpolarizations were not limited to neurones displaying an inflexion. The electrophysiological properties of the soma membrane of A delta neurones closely resembled those of A alpha neurones, while in several respects those of C neurones resembled the A beta I neuronal properties. The input resistance was found to be much greater in C than in A cells, although there was no significant difference between specific membrane resistance values calculated for the different groups. A number of A cells exhibited time-dependent rectification. PMID- 2987490 TI - Endothelium derived relaxant factor. PMID- 2987492 TI - Structural similarities between the plasma membrane binding sites for L-thyroxine and 3,3',5-triiodo-L-thyronine in cultured cells. AB - Using 125I-labeled L-thyroxine (T4), the binding of [125I]T4 to GH3 rat pituitary tumor cells was studied. At 15 degrees C, the binding of [125I]T4 to cells is saturable and specific. Least squares analysis of binding data showed two classes of binding sites with apparent dissociation constants of 4.3 +/- 0.3 nM and 350 +/- 30nM and binding capacities of (3.8 +/- 0.5) X 10(4) and (9.1 +/- 0.35) X 10(6) sites/cell, respectively. Affinity labeling of cells or purified plasma membranes with N-bromoacetyl-[125I]T4 (BrAc[125I]T4) showed a major specifically labeled protein band with an apparent molecular mass of 55 kilodaltons (kDal). Digestion of the 55-kDal protein from cells and plasma membrane by Staphylococcus aureus V8 protease or elastase gave similar peptide fragments. Thus, the 55-kDal protein labeled from intact cells is the same protein as that from purified plasma membranes. Peptide mapping was further used to compare the 55-kDal protein specifically labeled by either N-bromoacetyl-3,[125I]3',5-triiodo-L-thyronine (BrAc[125I]T3) or BrAc[125I]T4 in intact cells and highly purified plasma membranes. Very similar patterns were obtained. These results indicate that plasma membrane T3 and T4 binding sites have similar hormone binding domains. In addition the plasma membrane T3 and T4 binding sites of Swiss 3T3-4 mouse fibroblasts and A431 human epithelioid carcinoma cells are structurally similar to the T3 and T4 binding sites of GH3 cells. PMID- 2987491 TI - [Radiological aspects of cerebral and spinal cysticercosis. Apropos of 2 cases]. AB - Two cases of neurocysticercosis are reported, one cerebral and the other spinal. Results of radiologic imaging of this affection, a rare parasitosis in occidental Europe, are discussed with emphasis on computed tomography findings. In fact, when perfect conditions can be established the CT scan provides data confirming the diagnosis of cerebral cysticercosis in most cases. However, this scan is insufficient for investigation of ventricular or cisternal forms when gas or positive contrast is sometimes necessary for visualization of parasitic cysts. Radiologic diagnosis of extra- and intra-medullary forms is based an myelography with water-soluble contrast completed by a scan. PMID- 2987493 TI - Effect of median eminence lesions and hormonal replacement on the prolactin receptors in the adrenal gland and Langerhans islets from ovariectomized adult rats. AB - The effect of hyperprolactinemia induced by median eminence lesions (MEL) and ACTH and glucocorticoid replacement on prolactin (Prl) receptors was studied in the adrenal, isolated Langerhans islets and the liver. Adult rats were ovariectomized 15 days before MEL and they were divided in the following groups: 1) SHAM: injected with saline solution 3 times in alternate days; 2) MEL: saline solution; 3) MEL + ACTH: 50 micrograms: 10 IU/rat, s.c. (Synacthen) and 4) MEL + DEXA: 10 micrograms/rat (dexamethasone). For measuring total lactogenic binding sites an in vitro treatment of the membrane fraction with 4M MgCl2 was used. MEL originated a significant increase in Prl serum levels, which was not altered by injections of ACTH or dexamethasone. In contrast, serum corticosterone (B) levels in MEL rats were significantly lowered, and it was restored by ACTH. Unexpectedly, B levels increased when dexamethasone was administered to MEL rats. Prl receptors were diminished in the adrenal gland and Langerhans islets from MEL animals, as compared with the SHAM group. ACTH and glucocorticoid administration did not affect the pancreatic Prl receptors, while the adrenal gland exhibited a further lowering of Prl binding sites during ACTH treatment. Since no effect was found when dexamethasone was injected, a possible direct action of ACTH is suggested. On the other hand, Prl receptors were induced in the liver by MEL, and this action was abolished by dexamethasone and ACTH. Binding affinity in every tissue studied remained unchanged. Our data suggest that endogenous Prl is able to regulate its own receptors not only in the liver, but also in the adrenal gland and pancreatic islets. PMID- 2987494 TI - Absence of down-regulation of insulin receptors in human breast cancer cells (MCF 7) cultured in serum-free medium: comparison with epidermal growth factor. AB - MCF-7 cells were cultured either in RPMI-1640 medium containing 10% fetal calf serum (FCS) or in a serum-free (SF) medium supplemented with insulin, epidermal growth factor (EGF) and transferrin. Binding studies were performed with 125I insulin or 125I-EGF. In the FCS containing culture, down-regulation was seen for insulin receptors (47%), and for the EGF receptors (75%). Using cells grown in the serum-free medium we could not demonstrate down-regulation of the insulin receptors, while the EGF receptors were down-regulated to the same extent (74%). The number of binding sites per cell was about twice as much in cells cultured in FCS as that in SF medium. No significant differences were observed for receptor affinity of insulin or EGF in cells grown in both media. The rate of internalization of insulin or EGF into cells was similar in both culture conditions. The mechanism in which only EGF but not insulin demonstrated receptor down-regulation in SF medium remains unknown. PMID- 2987495 TI - Relationship between hCG receptors disappearance and steroid accumulation during long term hCG stimulation in porcine Leydig cell cultures. AB - LH-hCG receptors and steroidogenesis can be maintained for more than a week in porcine Leydig cell culture (chemically defined medium). Free receptors and steroid accumulation were measured following long time exposure to hCG. As expected, the percent of receptor disappearance increases with the hCG concentration in the incubation medium. However, the steroidogenesis was also proportional to hCG concentration for 24 hours as well as 48 hours. A 50% receptor disappearance was obtained for 0.5 ng hCG/ml which led to a steroid accumulation of 50% of the maximal accumulation obtained with 100 ng/ml. A very significative positive correlation (rho) was observed between receptor occupancy (internalized and/or irreversibly occupied receptors) and steroid accumulation (T and DHAS) during a 24 or 48 hour period (rho = 0.97 and 0.98 for T, and rho = 0.96 and 0.94 for DHAS). Following 24 or 48 hour exposure to increasing hCG concentration, the cells were acutely stimulated by 25 ng hCG/ml for 4 hours. The steroidogenic response obtained to this acute stimulation was inversely proportional to the concentration of hCG during the previous period. A negative correlation was found between receptor occupancy and acute steroidogenic response (rho : -0.96 and -0.98 for T). These results indicate that during long term stimulation as well as during acute restimulation the absolute number of occupied receptors conditioned the steroidogenic response. These results do not fit into the concept of spare receptors and suggest that in these immature cells all receptors are potentially active in the regulation of steroidogenesis. PMID- 2987497 TI - Adenoid cystic carcinoma of the vulva with lung metastases. A case report. AB - The presenting symptoms of adenoid cystic carcinoma of the vulva in a 57-year-old woman were a vulvar mass and pain. The mass was excised locally. The disease recurred in the lung four years later. The patient was treated with several regimens of chemotherapy, without significant effect. She died of pulmonary failure six years after diagnosis. PMID- 2987496 TI - Clinical, endocrine and ultrastructural study of XY gonadal dysgenesis. A case report. AB - About 120 cases of XY gonadal dysgenesis have been reported on. We treated such a patient with bilateral gonadectomy. The gonadal tissue's capacity to respond to hormonal trophic stimulation was assessed. When the gonads were examined ultrastructurally, structures with the morphologic characteristics of stromal ovarian cells, Sertoli's cells and Leydig's cells were found. Because of the potential malignancy of the XY gonads, bilateral gonadectomy and hormonal substitution therapy are recommended for these patients. We prefer to use combined hormone replacement with sequential estrogen and progesterone rather than sequential unopposed estrogen because of the small but increased risk of endometrial hyperplasia and carcinoma after long-standing sequential therapy. PMID- 2987498 TI - Unusual sarcoma arising in lymphoedema. PMID- 2987499 TI - Arthropod-borne virus zoonosis surveillance in the Cape Province: 1. Prospective serological investigations for virus activity in the Beaufort West and Middelburg Districts during 1981. AB - In addition to the routine sero-epidemiological surveillance for arthropod-borne viral zoonoses in the Cape Province carried out by the Department of Medical Microbiology and State Health Department of Virology laboratory, we conducted a prospective serological investigation for virus activity during 1981 in two districts of the Province, namely the Beaufort West and Middelburg districts, which experienced heavy rainfall during the first two months of that year. The approach used was to obtain paired serum samples from identified domestic stock representative of several species from 2-5 months apart and to test them for haemagglutination inhibition antibodies to Rift Valley fever, Wesselsbron and Middelburg virus antigen preparations in order to ascertain, as an indication of viral activity, whether changes in antibody levels occurred between the collection dates. The results indicated that there was probable activity of Rift Valley fever virus and activity of Wesselsbron virus (or related flaviviruses) and Middelburg virus (or related alphaviruses) in the Karoo between the middle of February and the end of July 1981. Despite this activity and heavy rainfall registered at meteorological stations in both the Beaufort West and Middelburg districts as well as general reports of heavy rainfall and considerable mosquito activity over widespread areas of the Karoo, Eastern and S.W. Cape Province, there were no epizootics or epidemics of overt arthropod-borne zoonotic viral disease in the province during 1981. PMID- 2987501 TI - Basolateral membrane potassium conductance is independent of sodium pump activity and membrane voltage in canine tracheal epithelium. AB - When secretagogues stimulate Cl secretion in canine tracheal epithelium, apical membrane Cl conductance (GCla) increases, and then basolateral membrane K conductance (GKb) increases. Conversely, inhibition of GCla results in a secondary decrease in GKb. The coordination of the two membrane conductances and regulation of GKb is critical for maintaining constant intracellular ion concentrations and transepithelial Cl secretion. The purpose of this study was to test two hypotheses about the regulation of GKb. First, we asked whether GKb is directly linked to the activity of the Na,K-ATPase. We found that pump activity could be dissociated from K conductance. Inhibition of the Na pump with ouabain, in nonsecreting tissues led to an increase in Gb. Elevation of the bathing solution K concentration produced a similar effect. Addition of ouabain to secreting tissues did not appear to alter Gb. These results indicate that GKb does not directly parallel Na pump activity. Second, we asked whether changes in GKb are voltage dependent. We prevented secretagogue-induced depolarization of the electrical potential difference across the basolateral membrane psi b by clamping psi b at its resting value during stimulation of Cl secretion with epinephrine. Despite maintaining psi b constant, the typical changes in transepithelial resistance and the ratio of membrane resistances persisted. This observation indicates that depolarization is not required for the secretagogue induced increase in GKb. In addition we examined the effect of depolarizing and hyperpolarizing psi b by passing transepithelial current in secreting and nonsecreting epithelia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987500 TI - Complex subcellular distributions of enzymatic markers in intestinal epithelial cells. AB - Current procedures for isolating intestinal epithelial cell surface and intracellular membranes are based on the assumption that each organelle is marked by some unique constituent. This assumption seemed inconsistent with the dynamic picture of subcellular organization emerging from studies of membrane turnover and recycling. Therefore, we have designed an alternative fractionation which is independent of a priori marker assignments. We subjected mucosal homogenates to a sequence of separations based on sedimentation coefficient, equilibrium density, and partitioning in aqueous polymer two-phase systems. The resulting distributions of protein and enzymatic markers define a total of 17 physically and biochemically distinct membrane populations. Among these are: basal-lateral membranes, with Na,K-ATPase enriched 21-fold; brush-border membranes, with alkaline phosphatase enriched as much as 38-fold; two populations apparently derived from the endoplasmic reticulum; a series of five populations believed to have been derived from the Golgi complex; and a series of five acid phosphatase rich populations which we cannot identify unequivocally. Each of the five enzymatic markers we have followed is associated with a multiplicity of membrane populations. Basallateral, endoplasmic reticulum, and Golgi membranes contain alkaline phosphatase at the same specific activity as the initial homogenate. Similarly, Na,K-ATPase appears to be associated with Golgi, endoplasmic reticulum, and brush-border membranes at specific activities two- to seven-fold that of the initial homogenate. PMID- 2987502 TI - Spin probe clustering in human erythrocyte ghosts. AB - A model has been developed for 5-nitroxide stearate, I(12,3), distribution in human erythrocyte ghosts which accurately predicts ESR spectral alterations observed with increased probe/total lipid (P/L) at 37 degrees C. This spin probe occupies a class of high-affinity, noninteracting sites at low loading. Saturation occurs with increasing probe concentration, and, at higher loading, the probe inserts itself at initially dilute sites to form membrane-bound clusters of variable size. No 'low' probe remains at high P/L where all I(12,3) clusters in a 'concentrated' phase. This model allows determination of the dilute/clustered probe ratio, and shows that I(12,3) segregates in erythrocytes at what might otherwise be considered low P/L (e.g., 1/359). These findings validate the earlier use of empirical parameters to estimate probe sequestration in biological membranes. PMID- 2987503 TI - Direct insertion and fluorescence studies of rhodamine-labeled beta-adrenergic receptors in cell membranes. AB - Previous studies utilizing the fluorescence of propanolol as a probe for the beta adrenergic receptor showed that this receptor is motionally constrained. To further study the beta-adrenergic receptor in situ we have reinserted rhodamine labeled beta-receptors into cell membranes. This report presents documentation of their insertion and physiologic viability. Beta-receptors were purified by affinity chromatography (10,000-fold), then fluorescently labeled with tetramethyl rhodamine isothiocyanate, repurified (55,000-fold) and incubated with rat pancreatic islet cells. The binding of 3H-dihydroalprenolol by these cells was increased from a Bmax of 168 +/- 2 to 309 +/- 20 fmol/mg protein with no change in Kd. Various treatments which remove peripheral membrane proteins, e.g. NaOH, lithium diiodosalicylate, and trypsinization, did not alter binding by the cells with inserted receptors indicating that the receptors inserted into cell membranes. In islet cells treated with Koshland's reagent I, beta-adrenergic binding was completely abolished, but following incubation with isolated beta receptors, the cells bound beta-adrenergic radioligand with a Bmax of 100 fmol/mg protein, indicating functionality on the part of the inserted receptors. Furthermore, insertion of isolated receptors into frog erythrocytes resulted in increased production of cAMP in response to added isoproterenol. In pancreatic islet cells, incubation with labeled receptors caused the fluorescence to shift in wavelength with increased intensity indicating a shift from an aqueous to a lipid environment, probably into the membrane. Using fluorescence (P), it was found that the inserted receptors became motionally constrained to a P of 0.38 (limiting Po = 0.42) during the first 15 min, remaining so for at least 2 hr. Colchicine (5 micrograms/ml) caused a decrease in P to 0.18 indicating release of constraint. Isoproterenol (10(-5)M) caused a rapid decrease to P = 0.15. This effect was blocked by propranolol. Propranolol itself (10(-5) M) had no effect. These results indicate that the labeled receptors rapidly insert into cell membranes and also support the view that agonist activation of the receptor causes an increase in receptor mobility, presumably due to release of constraint from cytoskeleton elements. PMID- 2987505 TI - Mapping of DNA gyrase cleavage sites in vivo oxolinic acid induced cleavages in plasmid pBR322. AB - We have developed a procedure which permits the mapping of DNA gyrase cleavage sites in vivo. Addition of oxolinic acid, an inhibitor of DNA gyrase, to growing cells of Escherichia coli containing the plasmid pBR322 resulted in double-strand cleavage of DNA, and allowed the isolation of significant quantities of linearized plasmid DNA after lysis of treated cells with sodium dodecyl sulfate. Initially the linear product was purified from agarose gels, cleaved by restriction endonucleases, and then subjected to Southern hybridization analysis using defined DNA probes. A number of distinct cleavage sites, used with varying degrees of efficiency, were identified within pBR322 using this simple procedure. To achieve greater resolution and to improve sensitivity, we then employed an electroblotting procedure to transfer DNA fragments from acrylamide gels onto nylon membranes. This alternative method does not require the isolation of the linearized product before performing the mapping procedure. The improved resolution obtained from acrylamide gels and the superior binding properties of the nylon membranes have allowed us to accurately map 74 distinct oxolinic acid induced cleavage sites within pBR322. The significance of these findings in light of previously reported studies in vitro, as well as the possible role of such sites during illegitimate recombination, are discussed. PMID- 2987504 TI - Interactions involving the cyanine dye, diS-C3-(5), cytochrome c and liposomes and their implications for estimations of delta psi in cytochrome c oxidase reconstituted proteoliposomes. AB - The interference of cytochrome c with absorption and fluorescence changes of the cyanine dye, diS-C3-(5), was investigated in the presence of liposomes and cytochrome c-oxidase reconstituted proteoliposomes. The apparent cytochrome c dependent quenching of diS-C3-(5) fluorescence, and the associated absorbance losses in the presence of liposomes and proteoliposomes in low ionic strength media, are due to destruction of the dye caused by cytochrome c-mediated lipid peroxidation. The rate of this reaction was further enhanced in the presence of hydrogen peroxide. Even in the absence of liposomes or proteoliposomes, a cytochrome c-induced breakdown of dye was observed in the presence of hydrogen peroxide. The cytochrome c mediated absorbance and fluorescence losses of diS-C3 (5) in liposomal or proteoliposomal systems are prevented by Ca2+ and La3+ ions, by ascorbate, by high ionic strength and by the antioxidant BHT. Under these conditions, the process of lipid peroxidation mediated by cytochrome c is inhibited either directly (e.g. by BHT) or indirectly, by preventing the binding of cytochrome c to lipid vesicles. The impact of these findings upon the experimental estimation of membrane potential in aa3-reconstituted proteoliposomes is discussed. PMID- 2987506 TI - A frameshift mutation at the junction of an IS1 insertion within lacZ restores beta-galactosidase activity via formation of an active lacZ-IS1 fusion protein. AB - The insertion of IS1 elements into lacZ results in the loss of beta-galactosidase activity, and such insertions exert a severe polar effect on the expression of the distal genes of the operon. In addition to these properties, the mutation lacZ::IS1-MS319 has the unique property of reversion to Lac+ (ts) spontaneously or after treatment with the frameshift mutagen ICR-191; such revertants retain the IS1 element. We have determined that the site of integration of IS1 into lacZ is at position 4338, 18 nucleotides from the end of the sequence encoding the C terminus of beta-galactosidase. Reversion to Lac+ promoted by ICR-191 results from the loss of a G residue from a GGG sequence located at the junction of lacZ and IS1. As a result an active, but temperature-sensitive, lacZ-IS1 fusion protein is formed containing six amino acids derived from IS1 which replace six amino acids encoded by lacZ. The IS1 element in MS319 is a new member of the iso IS1 family, which we designate IS1T. PMID- 2987507 TI - Members of the Amy-2 alpha-amylase gene family of mouse strain CE/J contain duplicated 5' termini. AB - Several members of the Amy-2 alpha-amylase multigene family from the CE/J strain of mouse have been cloned in cosmid vectors. Structural analysis of these recombinants reveals that the cloned Amy-2 copies contain tandem 5' termini. The duplicated 5'-terminal elements, which lie upstream from the Amy-2 cap site, are separated from their Amy-2 homologues by about 8000 bases. The orientation of these 5' orphons is the same as that of Amy-2. Gene titration and cloning experiments suggest that at least four of the approximately 15 Amy-2 copies present in the CE/J genome contain 5' orphon elements. The extent of sequence homology between 5' orphons and their gene homologues has been determined by DNA sequence analysis. All the orphons are identical and contain the entire 185 base pairs of the first exon, 49 base-pairs of the first intron and more than 400 base pairs of the Amy-2 5' flanking region. Intron and flanking-region sequences of the orphons differ by about 20% from their Amy-2 counterparts, and the exon by about 8%. The TATA box and the cap site are conserved, while the ATG translation initiation signal is mutated to ATA in the orphon. No transcription initiation has been detected at the orphon cap site using run-off transcription in isolated pancreatic nuclei in vitro. PMID- 2987508 TI - Helix movements and the reconstruction of the haem pocket during the evolution of the cytochrome c family. AB - Analysis of cytochromes c (tuna), c2 (Rhodospirillum rubrum), c550 (Paracoccus denitrificans) and c551 (Pseudomonas aeruginosa) shows that they contain 48 residues identifiable as homologous from superposition of the structures. The other 34 to 64 residues are in loops that vary greatly in sequence, length and conformation, or in alpha-helices that are found in only some of the structures. Of the 48 homologous residues, 17 are in three segments which pack onto the haem faces. In all four structures, these segments have the same conformations, and the same locations relative to the haem. The other 31 residues are in three alpha helices which are in contact with each other. These form the back and one side of the haem pocket. In cytochrome c551 the positions of the three alpha-helices have shifted and rotated, in comparison with cytochromes c and c2, by up to 5 A and 25 degrees relative to the haem. These shifts, facilitated by mutations at the helix helix interfaces, are related to the reconstruction of the propionic acid side of the haem pocket described by Almassy & Dickerson (1978). Together these effects produce alternative structures for the haem pocket. This mechanism of adaptation to mutation contrasts with that observed in the globins. In the globins, mutations also produce changes in helix interfaces and shifts of packed helices, but in the globins these shifts are coupled to conserve the structure of the haem pocket. PMID- 2987509 TI - Construction of plasmids carrying lacI mutations. PMID- 2987510 TI - Tn5 transposes independently of cointegrate resolution. Evidence for an alternative model for transposition. AB - The replicon fusion model for transposition predicts that each transposable element encodes a site-specific recombination activity (resolvase). A series of experiments were performed to test for the presence of such a resolvase in Tn5. None could be found when assays were devised to detect inter- or intramolecular recombination. An experiment was performed to determine if expression of resolvase occurs simultaneously with the transposition process, thus explaining the undetectable levels of site-specific recombination. In this case, no resolvase activity could be detected in cells that had undergone transposition. Instead, rearrangements were found that could be explained best by an alternative model of transposition. The rearrangements were consistent with a pathway that invokes excising the transposon from its initial site of insertion, followed by circularization of the element before its eventual transposition. The excision event may occur either with or without DNA synthesis. PMID- 2987512 TI - Simian virus 40 early promoter mutations that affect promoter function and autoregulation by large T antigen. AB - A set of nine mutants containing point mutations, and small deletions or insertions, were constructed in the early promoter region of simian virus 40 (SV40) to determine the role of the DNA sequences between the TATA box and the six upstream G + C-rich clusters in early transcription. The mutant templates were tested for transcription activity in vitro in HeLa cell extracts and in vivo in CV-1 and COS cells using the chloramphenicol acetyl transferase gene (CAT) assay. Both in vitro and in vivo results show that the narrow region from nucleotide positions 38 to 41 is an important domain of the early promoter. Deletion and insertion mutations most strongly affect the level of transcription. Specifically a four base-pair deletion in the promoter region enhances the level of transcription four- to sixfold in vitro, but causes a fourfold suppression of CAT gene expression in the in vivo assay. These opposite effects may result from changes in spacing under in vitro and in vivo conditions between the TATA box and the G + C-rich motifs where transcription factors may make simultaneous contact. Of the three T antigen binding sites (I, II and III), sites I and II have already been shown to be involved in the autoregulation of early transcription. Our mutational analyses demonstrate the role of site III, which partially overlaps with nucleotide positions 38 to 41, in the autoregulation of the SV40 early promoter. PMID- 2987511 TI - Analogs of cyclic AMP that elicit the biochemically defined conformational change in catabolite gene activator protein (CAP) but do not stimulate binding to DNA. AB - We have measured the effects on catabolite gene activator protein (CAP) of 22 synthetic analogs of cAMP. Each analog was assayed to test three parameters: (1) binding to CAP; (2) induction of the conformational change in CAP; and (3) activation of transcription. Thus we have identified seven cAMP analogs that bind to CAP as well or better than does cAMP, cause the assayed conformational change in CAP, yet exhibit no ability to activate transcription. We designate these analogs class D. The conformational change elicited in CAP by the class D analogs was further investigated by: (1) sensitivity to the proteolytic enzymes chymotrypsin, Staphylococcus aureus V8 protease, subtilisin and trypsin; (2) formation of inter-subunit covalent crosslinks by 5,5'-dithiobis(2-nitrobenzoic acid); and (3) degree of labeling of cysteine by [3H]N-ethylmaleimide. These experiments failed to detect a conformational difference between the CAP-class D and CAP-cAMP complexes. Filter binding and nuclease protection experiments indicate that the class D analogs do not efficiently support the binding of CAP to DNA. From these results, we suggest that there exists a hitherto undetected event dependent on cAMP, and required for CAP to bind to DNA. We suggest that this event involves a change that takes place in proximity to the N6 atom of cAMP. Three possible interpretations are discussed. PMID- 2987513 TI - Assignment of aromatic spin systems in the 1H nuclear magnetic resonance spectrum of adenylate kinase. AB - A combination of selective spin decoupling, two-dimensional double quantum spectroscopy, correlated spectroscopy (COSY), and pH titration experiments brought about the assignment of all tyrosyl spin systems and completed the assignment of the histidyl spin systems in porcine adenylate kinase. In the detection of the tyrosyl spin systems it proved to be advantageous to resort to the COSY method rather than to two-dimensional double quantum spectroscopy. In the titration experiments, His189 revealed a second apparent pK value at pH 8.3, which is explained by deprotonation of the adjacent residue Cys187. None of the seven tyrosyl side-chains shows any evidence for deprotonation up to the point of denaturation of the protein, which took place around pH 10. PMID- 2987514 TI - The effect of zinc on vitamin D3-induced cardiac necrosis. AB - Multifocal heart muscle necrotic lesions in rats were induced with high oral doses of vitamin D3 (300,000 iu/rat in three daily doses 100 000 iu each). The calcium content increased over 100 fold in the hearts of rats receiving vitamin D3. Parenteral pre-treatment with zinc sulphate (50 or 200 mg/rat in ten daily doses) resulted either in a reduction or in the total prevention of myocardial lesions on macroscopic, light and electron microscopic examination. The effect of zinc was dose-dependent. The administration of various doses of zinc sulphate resulted in a gradual normalization of heart calcium content. PMID- 2987515 TI - Influence of thyroid hormone levels on the electrical and mechanical properties of rabbit papillary muscle. AB - We have examined the influence of chronic in vivo alterations of thyroxine levels on the electrophysiological and mechanical properties of rabbit ventricular papillary muscles measured in vitro. Marked changes in the repolarization phase of the action potential and the time to peak tension of isometric twitches were observed when thyroid hormone levels were increased above or decreased below normal. The time to peak tension was consistently shorter than normal in hyperthyroid and longer than normal in hypothyroid preparations. In hypothyroid preparations the action potential duration was greater than that of controls at all stimulation frequencies tested (0.1 to 1.0 Hz). In hyperthyroid preparations, the repolarization phase consisted of an initial phase of fast repolarization to membrane potential values between -20 and -40 mV followed by a plateau. The early phase of repolarization persisted at all stimulation frequencies tested (0.1 to 1.0 Hz) but the plateau component increased markedly as the stimulation frequency was decreased. The early phase of repolarization was markedly reduced in the presence of 4-aminopyridine. These results suggest that thyroxine levels may modulate the kinetics of a transient outward current which in rabbit papillary muscle normally is responsible for the frequency dependence of action potential duration. Action potential amplitude, maximum rate of rise, resting membrane potential, and peak isometric twitch tension were not markedly different between the three classes of preparations. A second depolarizing response occurred in all hyperthyroid preparations at low stimulation frequencies (0.1 to 0.4 Hz) but not in control or hypothyroid preparations. This second depolarizing response, which was eliminated by D-600 treatment, was similar to the calcium-dependent slow action potentials recorded in other cardiac preparations. These two component action potentials could represent either intrinsic single cell activity, or a re entry wave of depolarization which results from nonhomogeneous excitation. PMID- 2987516 TI - Separation and reconstitution of regulatory and catalytic components of heart adenylate cyclase. AB - Ion-exchange chromatography of rabbit heart adenylate cyclase solubilized with lubrol PX results in two peaks of activity, AC I and AC II, differing in their sensitivity to guanylylimido-diphosphate [Gpp(NH)p], NaF and cholera toxin. AC I is activated 4- to 5-fold by Gpp(NH)p, 10- to 20-fold by NaF and 3- to 4-fold by cholera toxin. AC II is insensitive to Gpp(NH)p and cholera toxin, but is activated 2-fold by the fluoride. The differences in the regulatory properties of AC I and AC II are probably due to the unequal distribution of the N-protein, the regulatory component of adenylate cyclase, in the preparations. Addition of the N protein, obtained by thermoinactivation of AC I, to AC II results in restoration of the enzyme sensitivity to Gpp(NH)p as well as in an increase of the stimulating effect of NaF. It is shown that the lag-period observed during adenylate cyclase stimulation by Gpp(NH)p is due to the slow transition of the N protein into an activated conformation. This Gpp(NH)p-activated N-protein interacts with the catalytic component of adenylate cyclase without any lag period. Addition of GTP to the Gpp(NH)p-activated soluble adenylate cyclase complex leads to a decrease of the enzymatic activity. This process presumably occurs via substitution of Gpp(NH)p for GTP in the N-protein with subsequent hydrolysis of GTP and not by exchange of the N-protein-Gpp(NH)p complex for the N protein-GTP complex within the catalytic component of adenylate cyclase. PMID- 2987517 TI - Acute cerebellar syndrome in infectious mononucleosis: documentation of two cases with Epstein-Barr virus infection. AB - Acute cerebellar ataxia has been described occasionally with infectious mononucleosis. Two additional cases are reported with serologic identification of Epstein-Barr virus (EBV) infection in blood and cerebrospinal fluid. As with previously described cases, the outcome was benign, and examination and laboratory studies did not indicate diffuse neurologic involvement. Visual and brainstem auditory-evoked responses were normal. Electroencephalograms (EEG) demonstrated 14 and 6 per second positive spikes in both patients. This pattern is considered a normal variant and has been recorded from depth electrodes and reported with deep midline lesions. These cases support the prognosis of benign cerebellar involvement in infectious mononucleosis and suggest that evidence of EBV infection be sought in patients with acute ataxia. The significance of 14/sec and 6/sec positive EEG spikes is uncertain. PMID- 2987518 TI - Nucleotide and amino acid sequence coding for polypeptides of foot-and-mouth disease virus type A12. AB - The coding region for the structural and nonstructural polypeptides of the type A12 foot-and-mouth disease virus genome has been identified by nucleotide sequencing of cloned DNA derived from the viral RNA. In addition, 704 nucleotides in the 5' untranslated region between the polycytidylic acid tract and the probable initiation codon of the first translated gene, P16-L, have been sequenced. This region has several potential initiation codons, one of which appears to be a low-frequency alternate initiation site. The coding region encompasses 6,912 nucleotides and ends in a single termination codon, UAA, located 96 nucleotides upstream from a 3'-terminal polyadenylic acid tract. Microsequencing of radiolabeled in vivo and in vitro translation products identified the genome position of the major foot-and-mouth disease virus proteins and the cleavage sites recognized by the putative viral protease and an additional protease(s), probably of cellular origin, to generate primary and functional foot-and-mouth disease virus polypeptides. PMID- 2987519 TI - Intracellular synthesis of human parainfluenza type 3 virus-specified polypeptides. AB - The intracellular synthesis of human parainfluenza type 3 virus-specified polypeptides was examined by polyacrylamide gel electrophoresis of [35S]methionine-labeled cell extracts under reducing conditions. All of the virion structural proteins were detected in cell extracts, including: L, 180,000 molecular weight (180K); P, 83K; HN, 69K; NP, 66K; F0, 60K; F1, 51K; and M, 38K. P and NP were phosphorylated. HN and F were glycosylated. The kinetics of intracellular viral protein synthesis did not detect any early or late proteins. Pulse-chase experiments failed to detect any precursor-product relationships. No nonstructural proteins were detected. PMID- 2987520 TI - Two major outer envelope glycoproteins of Epstein-Barr virus are encoded by the same gene. AB - Two major outer envelope glycoproteins of Epstein-Barr virus, gp350 and gp220, are known to be encoded by 3.2- and 2.5-kilobase RNAs which map to the same DNA fragment (M. Hummel, D. Thorley-Lawson, and E. Kieff, J. Virol. 49:413-417). These RNAs have the same 5' and 3' ends. The larger RNA is encoded by a 2,777 base DNA segment which is preceded by TATTAAA, has AATAAA near its 3' end, and contains a 2,721-base open reading frame. The smaller RNA has one internal splice which maintains the same open reading frame. Translation of the 3.2- and 2.5 kilobase RNAs yielded proteins of 135 and 100 kilodaltons (Hummel et al., J. Virol. 49:413-417). The discrepancy between the 907 codons of the open reading frame and the 135-kilodalton size of the gp350 precursor is due to anomalous behavior of the protein in gel electrophoresis, since a protein translated from most of the Epstein-Barr virus open reading frame in Escherichia coli had similar properties. Antisera raised in rabbits to the protein expressed in E. coli specifically immunoprecipitated gp350 and gp220, confirming the mapping and sequencing results and the translational reading frame. The rabbit antisera also reacted with the plasma membranes of cells that were replicating virus and neutralized virus, particularly after the addition of complement. This is the first demonstration that the primary amino acid sequence of gp350 and gp220 has epitopes which can induce neutralizing antibody. We propose a model for the gp350 protein based on the theoretical analysis of its primary sequence. PMID- 2987521 TI - Cervical papillomaviruses segregate within morphologically distinct precancerous lesions. AB - We combined molecular hybridization and histological analysis of 58 biopsy samples from the uterine cervix to correlate the presence of certain strains of human papillomaviruses (HPVs) with specific pathological changes. HPV sequences were frequently detected (85%) in biopsies that contained a broad spectrum of precancerous changes. The presence of HPV 16 correlated strongly with precancers that exhibited morphological aberrations commonly found in invasive carcinomas and precancers that have a high risk of progression. This association underscores the potential importance of HPV type 16 in the genesis of genital cancers and emphasizes the value of histological analysis for detecting potentially oncogenic HPVs. PMID- 2987522 TI - Intracellular transport of herpes simplex virus gD occurs more rapidly in uninfected cells than in infected cells. AB - A mouse L cell line which expresses the herpex simplex virus type 1 immediate early polypeptides ICP4 and ICP47 was cotransfected with a cloned copy of the BglII L fragment of herpes simplex virus type 2, which includes the gene for gD, and the plasmid pSV2neo, which contains the aminoglycosyl 3'-phosphotransferase (agpt) gene conferring resistance to the antibiotic G418. A G418-resistant transformed cell line was isolated which expressed herpes simplex virus type 2 gD at higher levels than were found in infected cells. The intracellular transport and processing of gD was compared in transformed and infected cells. In the transformed Z4/6 cells gD was rapidly processed and transported to the cell surface; in contrast, the processing and cell surface appearance of gD in infected parental Z4 cells occurred at a much slower rate, and gD accumulated in nuclear membrane to a greater extent. Thus, the movement of HSV-2 gD to the cell surface in infected cells is retarded as viral glycoproteins accumulate in the nuclear envelope, probably because they interact with other viral structural components. PMID- 2987523 TI - Genomic expansion of Marek's disease virus DNA is associated with serial in vitro passage. AB - An EcoRI restriction endonuclease pattern of Md11 virus DNA, a very virulent strain of Marek's disease virus (MDV), was obtained by using total cellular DNA from infected cells. With the EcoRI restriction endonuclease pattern and a published BamHI map of MDV (Fukuchi et al., J. Virol. 51:102-109), we constructed a partial EcoRI map of a series of MDV clones (gift from H. J. Kung). The clones were used to identify a region of the Md11 genome which is altered as the oncogenic virus is passaged in vitro. This region was mapped into a 1.8-kilobase segment in the inverted-repeat sequences flanking the long unique region of the virus genome. The alteration appeared to result from multiple DNA insertions that produced an increase of 0.6 to 5.4 kilobases. Although the expansion of this region did not diminish the ability of MDV to replicate in vitro, it may be associated with the loss of Marek's disease oncogenicity. PMID- 2987524 TI - Phosphorylation sites on phosphoprotein NS of vesicular stomatitis virus. AB - The phosphoprotein NS of vesicular stomatitis virus which accumulates within the infected cell cytoplasm is phosphorylated at multiple serine and threonine residues (G. M. Clinton and A. S. Huang, Virology 108:510-514, 1981; Hsu et al., J. Virol. 43:104-112, 1982). Using incomplete chemical cleavage at tryptophan residues, we mapped the major phosphorylation sites to the amino-terminal half of the protein. Analysis of phosphate-labeled tryptic peptides suggests that essentially all of the label is within the large trypsin-resistant fragment predicted from the sequence of Gallione et al. (J. Virol. 39:52-529, 1981). A similar result has been obtained for NS protein isolated from the virus particle by C.-H. Hsu and D. W. Kingsbury (J. Biol. Chem., in press). Analysis of phosphodipeptides utilizing the procedures of C. E. Jones and M. O. J. Olson (Int. J. Pept. Protein Res. 16:135-142, 1980) enabled us to detect as many as six distinct phosphate-containing dipeptides. From these studies, together with the known sequence data, we conclude that the major phosphate residues on cytoplasmic NS protein are located in the amino third of the NS molecule and most probably between residues 35 and 106, inclusive. The studies also provide formal chemical proof that NS protein has a structure consistent with a monomer of the sequence of Gallione et al. as modified by J. K. Rose (personal communication). The low electrophoretic mobility of this protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis is not therefore due to dimerization. PMID- 2987525 TI - Use of simian virus 40 large T-beta-galactosidase fusion proteins in an immunochemical analysis of simian virus 40 large T antigen. AB - Simian virus 40 large T antigen is a multifunctional protein that is encoded by the early region of the viral genome. We constructed fusion proteins between simian virus 40 large T antigen and beta-galactosidase by cloning HindIII fragments A and D of the virus into the HindIII sites of expression vectors pUR290, pUR291, and pUR292. Large amounts of the fusion protein were synthesized when the DNA fragment encoding part of simian virus 40 large T antigen was in frame with the lacZ gene of the expression vector. Using Western blotting and a competition radioimmunoassay, we assessed the binding of existing anti-T monoclonal and polyclonal antibodies to the two fusion proteins. Several monoclonal antibodies reacted with the protein encoded by the fragment A construction, but none reacted with the protein encoded by the fragment D construction. However, mice immunized with pure beta-galactosidase-HindIII fragment D fusion protein produced good levels of anti-T antibodies, which immunoprecipitated simian virus 40 large T antigen from lytically infected cells, enabling derivation of monoclonal antibodies to this region of large T antigen. Therefore, the fusion proteins allowed novel epitopes to be discovered on large T antigen and permitted the precise localization of epitopes recognized by existing antibodies. The same approach can also be used to produce antibodies against defined regions of any gene. PMID- 2987526 TI - Analysis of tissue-specific methylation patterns of mouse mammary tumor virus DNA by two-dimensional Southern blotting. AB - We used a two-dimensional Southern blotting procedure to analyze the tissue specific methylation patterns of the five endogenous mouse mammary tumor viruses in the GR/A mouse strain. Our findings suggest that in certain tissues (brain, kidney, and liver) all proviruses are extensively methylated. In other tissues (spleen, placenta, and testes) all proviruses are hypomethylated to some degree. In these tissues individual proviruses display both quantitative and qualitative differences in methylation. We interpret the general patterns of tissue-specific hypomethylation in terms of a "hitch-hiker" model: mouse mammary tumor virus proviral methylation patterns reflect the tissue-specific activity of neighboring sequences. The observation that certain sites on particular proviruses are differentially methylated in a tissue-specific fashion may reflect tissue specific differences in the makeup or conformation, or both, of proviral containing chromatin. PMID- 2987527 TI - An immunoassay for the study of DNA-binding activities of herpes simplex virus protein ICP8. AB - An immunoassay was used to examine the interaction between a herpes simplex virus protein, ICP8, and various types of DNA. The advantage of this assay is that the protein is not subjected to harsh purification procedures. We characterized the binding of ICP8 to both single-stranded (ss) and double-stranded (ds) DNA. ICP8 bound ss DNA fivefold more efficiently than ds DNA, and both binding activities were most efficient in 150 mM NaCl. Two lines of evidence indicate that the binding activities were not identical: (i) ds DNA failed to complete with ss DNA binding even with a large excess of ds DNA; (ii) Scatchard plots of DNA binding with various amounts of DNA were fundamentally different for ss DNA and ds DNA. However, the two activities were related in that ss DNA efficiently competed with the binding of ds DNA. We conclude that the ds DNA-binding activity of ICP8 is probably distinct from the ss DNA-binding activity. No evidence for sequence specific ds DNA binding was obtained for either the entire herpes simplex virus genome or cloned viral sequences. PMID- 2987528 TI - Sequences in the polyomavirus DNA regulatory region involved in viral DNA replication and early gene expression. AB - We constructed and analyzed a series of deletion mutants in the noncoding regulatory region of tsa polyomavirus DNA to identify some of the sequences critical to the DNA replication origin and to the expression of the viral early genes in vivo. By using both transient and long-term assays under conditions where the influence of large T antigen (T-Ag) in replication or autoregulation was minimized, we observed no more than a 30% reduction in early gene expression upon removal of the CAAT or TATA elements or both. These assays demonstrated a predominant effect of upstream promoter or enhancer elements and indicated that removal of the CAAT or TATA boxes did not significantly affect viral early gene expression. Studies on the replicative ability of these mutants in mouse cells constitutively expressing the polyoma early proteins revealed that the removal of DNA sequences contained within a previously identified T-Ag high-affinity binding site (nucleotides 39 to 64) abolished viral DNA replication, whereas removal of two other high-affinity sites, closer to the early mRNA cap sites, did not. Furthermore, a deletion including this same high-affinity site plus a low affinity binding site within the 32-base-pair palindrome of the origin core sequences eliminated the ability of the viral large T-Ag to efficiently repress early gene transcription. It is thus possible that the origin-proximal high affinity T-Ag binding site is involved in both of the functions of large T-Ag, i.e., the initiation of viral DNA replication and the autoregulation of early gene transcription. PMID- 2987529 TI - DNA rearrangement in the control region for early transcription in a human polyomavirus JC host range mutant capable of growing in human embryonic kidney cells. AB - A human polyomavirus JC virus (JCV) host range mutant (JC-HEK) can grow in human embryonic kidney cells, whereas the brain cell-tropic wild-type JCV strain (Mad 1) cannot; JC-HEK contains two complementing defective DNAs, JC-HEK-A and JC-HEK B. We determined the nucleotide sequence of the putative transcriptional control region of JC-HEK-A DNA that can induce T-antigen synthesis in human embryonic kidney cells and compared it with the sequence of JCV Mad-1 DNA. The JC-HEK-A control region was found to have a complex DNA rearrangement, namely, a partial local duplication of a noncoding region generating two extra replication origins and translocation of segments from the large-T-antigen gene (415 base pairs) and the VP-1 gene (78 base pairs). In the rearranged segment, JC-HEK-A had seven sets of the sequence 5'TGGA(T)A(T)A(T)3', which is found in the simian virus 40 enhancer core, whereas JCV Mad-1 had only one set in its control region. JC-HEK-A also had a 5'TGGAAGTGTAA3' sequence resembling the adenovirus early region 1A enhancer core sequence 5'AGGAAGTGAA3'. Because the viral enhancer is host discriminatory and because another human polyomavirus, BK virus, that grows well in human embryonic kidney cells has these signals in its control region, it is likely that some of the newly acquired signals in JC-HEK play an important role in the altered host range of JCV. PMID- 2987531 TI - Production of soluble suppressor factors by herpes simplex virus-stimulated splenocytes from herpes simplex virus-immune mice. AB - Indirect evidence indicates that herpes simplex virus (HSV)-specific cytotoxic-T lymphocyte induction is regulated by suppressor cells. To search for such suppressor effects, supernatant fluids from splenocyte cultures from normal and HSV-immune mice cultured either with or without viral stimulation were tested for their ability to inhibit HSV-specific cytotoxic-T-lymphocyte induction. Only the supernatant fluid from the HSV-stimulated, HSV-immune cultures contained a suppressor activity (HSV-SF). HSV-SF was produced by nylon-wool-purified Thy 1+ cells. HSV-SF was detectable after 3 days of culture and would only suppress cytotoxic-T-lymphocyte induction if HSV-SF was added within 24 h of initiation of the test cultures. HSV-SF was neither dialyzable nor heat stable. Molecular sieve chromatography of HSV-SF yielded multiple peaks of suppressor activity. Although most of these peaks exhibited nonspecific suppressor activity, the suppression mediated by the 90,000 to 150,000-molecular-weight fractions was antigen specific and genetically restricted. These results provide direct evidence for the regulation of HSV-cytotoxic-T-lymphocyte induction by a novel suppressor factor. PMID- 2987530 TI - Nucleotide sequence analysis of a variant human T-cell leukemia virus (HTLV-Ib) provirus with a deletion in pX-I. AB - A variant of human T-cell leukemia virus subgroup I (HTLV-I), designated HTLV-Ib, has been isolated from a transformed T-lymphocytic cell line established from a Zairian patient with adult T-cell lymphoma. A recombinant phage clone of the variant provirus, denoted lambda MC-1, hybridizes under high stringency to HTLV-I DNA probes, but 17 of 43 restriction enzyme sites differ from those of HTLV-I, 10 of them clustering within 1.5 kilobases in the env-pX region. Since this variant virus retains its capacity to transform T-cells in vitro, and since a pX product is suspected to be important in transformation, we have determined the nucleotide sequence of the entire pX region of this virus for comparison to the prototype HTLV-I. In addition, the region between the gag and pol genes, parts of the pol and env genes, and a portion of the U3 region of the long terminal repeat sequence were also analyzed. We noted 141 single-base-pair changes among 3,897 base pairs, which were relatively well distributed over those portions of the provirus that were examined. In addition, an 11-base-pair deletion was found which included the potential initiator ATG codon of the first open reading frame of pX (pX-I). The next potential initiator codon predicted by the sequence is followed by 10 codons and then a termination codon. An identical deletion was also demonstrated in the only provirus present in another cell line established from the same patient on a different occasion after transformation in vitro of normal human umbilical cord blood cells. These results indicate that pX-I is not required for transformation. PMID- 2987532 TI - Generation of a recombinant Moloney murine leukemia virus carrying the v-src gene of avian sarcoma virus: transformation in vitro and pathogenesis in vivo. AB - A Moloney murine leukemia virus (M-MuLV) recombinant carrying the v-src gene of avian sarcoma virus was generated by the introduction of a cloned portion of v src from Schmidt-Ruppin A avian sarcoma virus into a molecular clone of M-MuLV provirus at the recombinant DNA level. The v-src sequences (lacking a portion of the 5' end of v-src) were inserted into the p30 region of the M-MulV gag gene so that M-MuLV gag and v-src were in the same reading frame. Transfection of this chimeric clone, pMLV(src), into NIH 3T3 cells which were constitutively producing M-MuLV gag and pol protein resulted in the formation of foci of transformed cells. Infectious and transforming virus could be recovered from the transformed cells. This virus was designated M-MuLV(src). M-MuLV(src)-transformed cells contained two novel proteins of 78 and 90 kilodaltons. The 78-kilodalton protein, p78gag-src, contained both gag and src determinants, exhibited kinase activity in an immune kinase assay, and is probably a fusion of Pr65gag and src. The 90 kilodalton protein, which is of the appropriate size to be the gPr80gag fused to src, contained gag determinants as well as a V8 protease cleavage fragment typical of the carboxy terminus of avian sarcoma virus pp60src. However, it could not be immunoprecipitated with an anti-v-src serum. M-MuLV(src)-transformed cells showed elevated levels of intracellular phosphotyrosine in proteins, although the elevation was intermediate compared with cells transformed with wild-type v-src. M-MuLV and amphotropic murine leukemia virus pseudotypes of M-MuLV(src) were inoculated into newborn NIH Swiss mice. Inoculated mice developed solid tumors at the site of inoculation after 3 to 6 weeks, with most animals dying by 14 weeks. Histopathological analysis indicated that the solid tumors were mesenchymally derived fibrosarcomas that were both invasive and metastatic. PMID- 2987533 TI - The alpha sequence of the cytomegalovirus genome functions as a cleavage/packaging signal for herpes simplex virus defective genomes. AB - Although herpes simplex virus (HSV) 1 and human cytomegalovirus (CMV) differ remarkably in their biological characteristics and do not share nucleotide sequence homology, they have in common a genome structure that undergoes sequence isomerization of the long (L) and short (S) components. We have demonstrated that the similarity in their genome structures extends to the existence of an alpha sequence in the CMV genome as previously defined for the HSV genome. As such, the alpha sequence is predicted to participate as a cis-replication signal in four viral functions: (i) inversion, (ii) circularization, (iii) amplification, and (iv) cleavage and packaging of progeny viral DNA. We have constructed a chimeric HSV-CMV amplicon (herpesvirus cis replication functions carried on an Escherichia coli plasmid vector) substituting CMV DNA sequences for the HSV cleavage/packaging signal in a test of the ability of this CMV L-S junction sequence to provide the cis signal for cleavage/packaging in HSV 1-infected cells. We demonstrate that the alpha sequence of CMV DNA functions as a cleavage/packaging signal for HSV defective genomes. We show the structure of this sequence and provide a functional demonstration of cross complementation in replication signals which have been preserved over evolutionary time in these two widely divergent human herpesviruses. PMID- 2987534 TI - Synthesis and processing of glycoprotein gG of herpes simplex virus type 2. AB - Monoclonal antibody 13 alpha C5-1-A11 immunoprecipitated two major polypeptides of molecular weights 108,000 and 120,000 from extracts of herpes simplex virus type 2-infected BHK-21 cells labeled with [35S]methionine or [3H]glucosamine. In pulse-chase experiments, both labels were chased from the 120,000-molecular weight peptide (120K peptide) into the 108K molecule. Endoglycosidase H (endo H) reduced the 120K peptide to a 112K peptide but did not affect the 108K peptide. Similar profiles were obtained with monoclonal antibody AP-1 which reacts with a 92K glycoprotein, gG, which maps to the short unique region of the genome. Cross absorption experiments indicated that both antibodies reacted with the same peptides, suggesting that the 120K peptide is a partially glycosylated high mannose-type precursor of gG (pgG1). Immunoprecipitation from monensin-treated cells indicated that pgG1(120K) may undergo peptide cleavage to form a 74K high mannose-type peptide (pgG2) and that this 74K peptide may be further processed into an endo H-resistant 110K to 116K peptide. In the presence of tunicamycin, gG(108K) was replaced by 110K and 105K peptides which were resistant to both endo H and endoglycosidase F. The 105K peptide was the only molecule labeled by [3H]galactose or [3H]glucosamine in the presence of tunicamycin, and none of the peptides were labeled with [3H]mannose, indicating the probable presence of O linked sugars in the 105K peptide. Our results imply that cotranslational glycosylation of the unglycosylated precursor 110K peptide results in the high mannose-type pgG1(120K), which probably undergoes peptide cleavage. This putative cleavage product may then mature into gG (108K) by the trimming of sugars and the addition of complex and probably O-linked sugars; the high-mannose-type pgG2(74K) is probably an intermediate peptide formed in this process. PMID- 2987535 TI - Expression of influenza virus NS2 nonstructural protein in bacteria and localization of NS2 in infected eucaryotic cells. AB - The nonstructural NS2 protein of influenza A/PR/8/34 virus was efficiently expressed in bacteria, and monospecific antisera were prepared against the bacterially synthesized polypeptide. These antisera were cross-reactive among the NS2 proteins of various influenza A viruses. However, they did not react with the NS2 of influenza B/Lee/40 virus nor with other proteins of influenza A viruses such as NS1. Antisera against NS2 were used to determine that the NS2 protein is localized in the cell nucleus during influenza virus infection, as shown by immunofluorescence microscopy. Cells infected with simian virus 40 recombinants containing the influenza virus NS gene revealed that both the NS1 and NS2 proteins appeared in the nucleus, even in the absence of expression of other influenza virus-specific components. PMID- 2987536 TI - Trypsin sensitivity of the Sabin strain of type 1 poliovirus: cleavage sites in virions and related particles. AB - Treatment of the Sabin strain of type 1 poliovirus with trypsin produced two stable fragments of capsid protein VP1 which remained associated with the virions. Trypsinized virus was fully infectious and was neutralized by type specific antisera. The susceptible site in the Sabin 1 strain was between the lysine at position 99 and the asparagine at position 100. A similar tryptic cleavage occurred in the Leon and Sabin strains of type 3 poliovirus, probably at the arginine at position 100, but not in the type 1 Mahoney strain, which lacks a basic residue at either position 99 or position 100. Tryptic treatment of heat treated virus and 14S assembly intermediates produced unique stable fragments which were different from those produced in virions. The implications of our results for future characterization of the surface structures of these particles and structural rearrangements in the poliovirus capsid are discussed. PMID- 2987537 TI - Phytohemagglutinin activation of the transcription of the bovine leukemia virus genome requires de novo protein synthesis. AB - Addition of supramitogenic doses of phytohemagglutinin (PHA) to short-term cultures of neoplastic or nonneoplastic lymphocytes infected with bovine leukemia virus increased the synthesis of the major core virion antigen (p25) by 5- to 10 fold. Such stimulation was not due to the mitogenic effect of PHA or to a generalized increase in cellular RNA or protein synthesis but rather to enhanced transcription of the viral genome by a PHA-induced protein. PMID- 2987538 TI - Insertional activation of c-myc by reticuloendotheliosis virus in chicken B lymphoma: nonrandom distribution and orientation of the proviruses. AB - Chicken syncytial virus, a member of the reticuloendotheliosis virus family, can induce chicken B lymphomas indistinguishable from those caused by avian leukosis virus. Previously, we have demonstrated that the chicken syncytial virus proviruses in these tumors are linked to the proto-oncogene c-myc. We have now determined the arrangement of chicken syncytial virus proviruses in 22 tumors. The results indicate that these proviruses, without exception, are integrated upstream from the second c-myc exon. At least 70% of these insertion sites are clustered in a 0.5-kilobase region immediately preceding the exon. The proviruses are all arranged in the same transcriptional orientation as c-myc. This type of provirus organization bears strong resemblance to that of the avian leukosis virus proviruses involved in c-myc activation. PMID- 2987539 TI - Effects of cobra venom factor treatment on latent feline leukemia virus infection. AB - The role of the complement system in containment of feline leukemia virus infection was studied by cobra venom factor treatment of feline leukemia virus immune cats. One to three weeks after cobra venom factor treatment, an increase in viral antigen in marrow myelomonocytic cells and circulating immune complexes was noted. Prevention of reactivation of feline leukemia virus infection may in part depend on an intact complement system. PMID- 2987540 TI - Simian virus 40 small-t antigen-induced theophylline resistance is not mediated by cyclic AMP. AB - Small-t antigen of simian virus 40 renders CV-1 cells resistant to growth arrest induced by theophylline and other methylxanthines. Elevated levels of cyclic AMP are not involved in growth arrest of CV-1 cells by methylxanthines, and small-t antigen does not alter cyclic AMP levels dramatically after infection. PMID- 2987541 TI - The immediate-early enhancer element of herpes simplex virus type 1 can replace a regulatory region of the c-Ha-ras1 oncogene required for transformation. AB - A 0.8-kilobase SacI DNA fragment in the distal 5'-noncoding region of the c-Ha ras1 oncogene hybridized to high guanine X cytosine sites of herpes simplex virus type 1 (HSV-1) DNA restriction fragments. Nucleotide sequence comparisons localized one of these sites to the intergenic region of HSV between the immediate-early genes coding for IEmRNA-3 and IEmRNA-4/5 that has enhancer-type activity. We tested the possibility that the HSV-1 enhancer and the upstream c-Ha ras1 SacI fragment were functionally related by assaying for the capacity of recombinant plasmids in which the HSV-1 enhancer replaced the oncogene 0.8 kilobase SacI fragment to transform NIH/3T3 cells. Deletion of the 0.8-kilobase SacI fragment abolished the biological activity of c-Ha-ras1, but its replacement by the HSV-1 enhancer fully restored it. These results confirm the enhancer properties of the HSV-1 immediate-early intergenic region and suggest that c-Ha ras1 sequences contained within the 0.8-kilobase SacI fragment plays a role in the transcriptional activation of the oncogene. PMID- 2987542 TI - DNA sequence of the leftward junction in the adenovirus-simian virus 40 hybrid Ad2+D2 and determination of the structure of the D2-T antigen. AB - The nucleotide sequence of the junction between the simian virus 40 early region and the adenovirus type 2 late region L4 in the hybrid virus Ad2+D2 was determined. The deduced amino acid sequence suggests that the D2-T antigen is a chimeric protein sharing 594 amino acids with the C-terminal end of the simian virus 40 T antigen and 104 amino acids with the N terminus of the adenovirus type 2 33,000-molecular-weight protein. The predicted structure of the D2-T antigen was confirmed by an immunoprecipitation analysis. PMID- 2987543 TI - Nucleotide sequence analysis of endogenous murine leukemia virus-related proviral clones reveals primer-binding sites for glutamine tRNA. AB - Nucleotide sequences of the region that corresponds to the site of tRNA primer binding for a functional retrovirus were determined in five murine leukemia virus related sequence clones from mouse chromosomal DNA, which contain a unique 170 to 200-base-pair additional internal segment in the long terminal repeats. The 3' terminal 18-nucleotide sequence of a major glutamine tRNA isoacceptor was found to match well with the putative primer binding site: 18 of 18 in three clones, 17 of 18 in one clone, and 16 of 18 in one clone. This implies that most of these endogenous proviral sequences of the mouse genome, if replicated as retroviruses, will be different from ecotropic murine leukemia viruses and most mammalian type C retroviruses in using glutamine tRNA, rather than proline tRNA, as a primer. PMID- 2987544 TI - Role of the 3' long open reading frame region of bovine leukemia virus in the maintenance of cell transformation. AB - Viral RNA expression was studied by dot blot hybridization with polyadenylated RNAs extracted from a bovine (YR-1) and an ovine (YR-2) tumor cell clone. Both clones were derived from in vivo bovine leukemia virus-induced tumors. The probes used were either the bovine leukemia virus information or only the long open reading frame sequences. No viral RNA corresponding to the bovine leukemia virus long open reading frame region was detected in YR-2, and a very limited amount of bovine leukemia virus messages was unraveled in YR-1. These results strongly suggest that viral expression, even in the long open reading frame region, is not required to maintain transformation of at least some tumor cells. PMID- 2987545 TI - Wilms tumor in horseshoe kidneys: a report from the National Wilms Tumor Study. AB - An analysis of information on 2,961 patients with Wilms tumor entered in the National Wilms Tumor Study up to July 1983 revealed 13 tumors occurring in horseshoe kidneys. The correct preoperative diagnosis was made in 6 of the 13 patients. At presentation 5 patients had stage 1, 3 stage 2 and 5 stage 3 disease. Major preoperative or intraoperative tumor spillage occurred in 3 patients. The over-all survival rate was 85 per cent after a mean followup of 45 months. The surgical, pathological and therapeutic aspects of Wilms tumor in horseshoe kidneys were analyzed carefully. The frequency of horseshoe kidneys in patients with Wilms tumor registered with the National Wilms Tumor Study was determined and was used in conjunction with population data to estimate the relative risk of Wilms tumor occurrence in children with horseshoe kidneys. PMID- 2987546 TI - Serologic surveillance for vesicular stomatitis virus on Ossabaw Island, Georgia. AB - Seventeen species of mammals and seven species of birds from Ossabaw Island, Georgia, were tested for vesicular stomatitis (VS) neutralizing antibodies. Seropositive results were restricted to mammals with six of 17 species testing seropositive for VS (New Jersey type) neutralizing antibodies. Seropositive species included: raccoons (Procyon lotor), white-tailed deer (Odocoileus virginianus), feral swine (Sus scrofa), cattle (Bos taurus), horses (Equus caballus), and donkeys (Equus asinus). All tests for VS (Indiana type) were negative. PMID- 2987548 TI - Poxvirus infection of the Manx shearwater (Puffinus puffinus). AB - During studies on the etiology of puffinosis, a disease of the Manx shearwater, 1 to 4% of full-grown birds were found to have dry, non-pigmented lesions on the webs of the feet. Poxvirus infection was detected in six of seven full-grown birds with such lesions. The lesions contained large encapsulated inclusions which were packed with mature and immature poxvirus particles. Poxvirus infection was not apparent in shearwater fledglings during puffinosis epizootics, and its spatial distribution was not related to that of puffinosis. The results indicate that poxvirus infection produces a mild, self-limiting disease in shearwaters and is not the cause of puffinosis. PMID- 2987547 TI - Persistence of inclusion body disease of cranes virus. AB - Persistence of inclusion body disease of cranes virus (IBDCV) was determined by monitoring virus shedding, serum antibody and in vitro cultivation of trigeminal ganglia from cranes. Samples were collected from captive cranes surviving the outbreak in 1978 and from cranes inoculated with the virus. Tissues and fluids from eggs of cranes that survived the outbreak were also tested for virus. Latent IBDCV was found in the trigeminal ganglion of one crane that was exposed to the virus in 1978. Spontaneous or induced (cyclophosphamide and dexamethasone) reactivation of viral shedding was not detected in any cranes tested. Five of six experimentally inoculated cranes died with lesions of an inclusion body disease, but virus was isolated from only three of them. One crane shed detectable levels of IBDCV prior to death. The surviving crane developed a transient antibody response without evidence of viral shedding, after five exposures to the virus. A latent infection was not detected in this crane. Serum antibody titers of cranes that survived the outbreak declined from 1980-1982. No virus was isolated from the eggs. Although IBDCV is capable of persisting in a latent form in the trigeminal ganglia of cranes, the low frequency of viral shedding suggests that this virus may be only a sporadic problem. PMID- 2987549 TI - Cutaneous fibromas of moose (Alces alces). PMID- 2987550 TI - Modoc-like virus isolated from wild deer mice (Peromyscus maniculatus) in Alberta. AB - Small mammals were trapped in northeastern Alberta, Canada during 1976. Blood samples from these animals were tested for virus by inoculation of suckling mice. Blood clots from two deer mice yielded isolates of the same virus. The virus was related antigenically to a number of flaviviruses which have been isolated from mammals in Central America and North America and was related most closely to Modoc virus. Physical, chemical, and biological properties of the virus were similar also to those of Modoc virus. It did not produce illness or death in deer mice inoculated in the laboratory. Neutralization tests indicated that 1/38 (3%) red squirrels (Tamiasciurus hudsonicus), 3/35 (9%) least chipmunks (Eutamius minimus), 13/109 (12%) deer mice, and 3/50 (6%) humans were infected naturally. This is the first reported evidence of infection of red squirrels and chipmunks with a Modoc-like virus. These data extend the range of Modoc-like viruses northward by 1,500 km and comprise the first isolate from mammals in the boreal forest of Canada. PMID- 2987551 TI - Lung cancer cell type and asbestos exposure. AB - It is often claimed that adenocarcinoma is the most frequent type of lung cancer seen in workers exposed to asbestos. Careful review of the published data, however, fails to support this claim. Rather, it seems that all types of lung cancers occur in asbestos workers, and the presence or absence of a specific cell type cannot be used to prove or disprove an association of carcinoma and asbestos exposure. PMID- 2987552 TI - Disseminated CMV infection. PMID- 2987554 TI - Not there yet, but "on our way" in AIDS research, scientists say. PMID- 2987553 TI - Antibodies to AIDS-associated retrovirus distinguish between pediatric primary and acquired immunodeficiency diseases. AB - Antibody to acquired immunodeficiency syndrome (AIDS)-associated retroviruses (ARVs) was investigated in 68 pediatric patients with abnormalities of T-cell and/or B-cell immunity. All except seven patients conformed to a specific World Health Organization classification for immunodeficiency disease. These seven patients had polyclonal hypergammaglobulinemia and T-cell immunodeficiency. Six of the seven patients had antibody to ARV and had risk factors associated with AIDS. The one patient without antiviral antibody had no AIDS risk factors. No antibody was detected in 61 patients with other primary immunodeficiency disorders. We conclude that ARV first appeared in our population of immunodeficient pediatric patients prior to 1978, is associated with a distinctive immunologic phenotype consisting of polyclonal hypergammaglobulinemia and T-cell immunodeficiency, and does not appear as an opportunistic infection in other immunodeficiency disorders. Detection of the retrovirus associated with AIDS is of value in identifying infants and children who may have unique medical and social problems that occur with AIDS. PMID- 2987555 TI - More heterosexual spread of HTLV-III virus seen. PMID- 2987556 TI - Neurological complications appear often in AIDS. PMID- 2987557 TI - Screening for HTLV-III antibodies: the relation between prevalence and positive predictive value and its social consequences. PMID- 2987558 TI - AIDS serology testing in low- and high-risk groups. AB - The performance characteristics of the acquired immunodeficiency syndrome (AIDS) retrovirus serological tests including enzyme-linked immunosorbent assay (ELISA), Western blot, and immunofluorescence assay were defined in a clinical laboratory setting by testing 1,257 serum specimens from low- and high-risk groups for AIDS. The three prototype AIDS retroviruses (lymphadenopathy-associated virus, human T lymphotropic virus III, and AIDS-associated retrovirus) were equally suitable as target antigen for these assays. Sera from six of 74 laboratory and health care personnel and 91 of 1,014 unselected blood donors were falsely positive by ELISA (positive to negative ratio [P/N], greater than or equal to 2) based on the lack of Western blot confirmation. Only two true-positives (two [0.2%] of 1,014 blood donors) were detected in these low-risk groups. In contrast, 106 of 108 specimens with ELISA P/N ratios of 2 or greater from the high-risk groups including asymptomatic homosexual men, hemophiliacs, AIDS-related complex patients, and AIDS patients were positive by Western blot and immunofluorescence assay. Four false-negative ELISA results based on positive immunofluorescence assay and Western blot were found in the AIDS patient group. Ten of 69 AIDS patients were negative by all three serological tests. The consequence of maintaining high sensitivity for the ELISA (P/N ratio, greater than or equal to 2) as a screening test was a loss of specificity. The number of false-positive results necessitated the use of a confirmation test with greater specificity. PMID- 2987560 TI - Simultaneous occurrence of Pneumocystis carinii pneumonia, cytomegalovirus infection, Kaposi's sarcoma, and B-immunoblastic sarcoma in a homosexual man. AB - The most common manifestations of the acquired immunodeficiency syndrome include Pneumocystis carinii pneumonia and/or Kaposi's sarcoma. High-grade B-cell lymphomas have also been reported in homosexual men at risk for the acquired immunodeficiency syndrome. We herein present the case of a homosexual man, who presented simultaneously with Pneumocystis carinii pneumonia, acute cytomegalovirus infection, Kaposi's sarcoma, and B-cell immunoblastic sarcoma. Severe compromise of both the B- and T-cell arms of the immune system was documented. The patient had evidence of exposure to the human T-lymphotropic retrovirus III, evidence of reactivation of Epstein-Barr virus infection, and cytomegalovirus inclusions within Kaposi's sarcoma tissue. We conclude that exposure to these viral agents in the setting of severe immunocompromise may have led to the observed "opportunistic" neoplasms. PMID- 2987559 TI - Human T-lymphotropic retrovirus type III/lymphadenopathy-associated virus antibody. Association with hemophiliacs' immune status and blood component usage. AB - We studied the human T-lymphotropic retrovirus type III/lymphadenopathy associated virus (HTLV-III/LAV) antibody status of 234 factor VIII concentrate recipients, 36 factor IX concentrate recipients, 69 long-term recipients of frozen packed red blood cells, and 47 persons not receiving routine transfusion therapy. Factor VIII concentrate recipients had a significantly higher rate of seropositivity (74%) than any other group. Factor IX concentrate recipients had a significantly higher rate (39%) than recipients of frozen packed red blood cells (4%) or nontransfused persons (4%). In factor VIII concentrate recipients, HTLV III/LAV seropositivity was significantly associated with more severe hemophilia, greater factor dosage, elevated immunoglobulin and immune complex levels, lower T helper lymphocyte numbers, and lower ratios of T-helper to T-suppressor lymphocytes. For factor IX concentrate recipients, seropositivity was associated with more severe hemophilia. Antibody-positive factor IX concentrate recipients had a lower rate of seropositivity to HTLV-III/LAV p41 membrane antigen than did antibody-positive factor VIII concentrate recipients, but factor VIII and factor IX concentrate recipients had similar rates of seropositivity to core antigens. We conclude that both factor VIII and factor IX concentrates may transmit HTLV III/LAV. For factor VIII recipients, HTLV-III/LAV seropositivity is associated with altered immune test results. PMID- 2987561 TI - Sequelae of breast irradiation. PMID- 2987562 TI - Immunodeficiency. PMID- 2987563 TI - [Effects of diltiazem on plasma ACTH, beta-ELI and ADH levels during surgery]. PMID- 2987565 TI - [The clinicopathological effects of infusion of cis-platinum into the bronchial artery on lung cancer]. AB - Ten patients with histologically and/or cytologically confirmed lung cancer were treated with cis-platinum (CDDP) infused into the bronchial artery (BAI). Radiological findings on chest X-rays taken two weeks after BAI were evaluated according to Takeuchi's criteria. The radiological results were judged to be quite effective in one case of squamous cell carcinoma, with more than 50% shrinkage of the tumor shadow, and to be effective in nine others, with more than 20% shrinkage. As for the histological effects, grade III in one case of squamous cell carcinoma and grade IIa in two cases of adenocarcinoma and one of large cell carcinoma were obtained by Shimosato's criteria. In conclusion, BAI with CDDP was sufficiently useful as a chemotherapeutic modality for lung cancer, especially squamous cell carcinoma, because higher local dosage of CDDP can be delivered without systemic toxicity. PMID- 2987564 TI - [Pre-synaptic inhibition and pentobarbital]. PMID- 2987566 TI - [A long-term survivor of small cell lung cancer]. AB - A 60-year-old man with small cell lung cancer who has been alive seven years in good condition after the combined therapy with radiation and anticancerous drugs, is presented. Chest X-ray examinations revealed the right middle lobe atelectasis and lymph node enlargement of the right hilum. Small cell carcinoma (oat cell type) was diagnosed by cytological examinations of the sputum. The patient was given 42 Gy of telecobalt, one course of METT as chemotherapy and PSK (3.0 g X 52) and BCG-CWS (300 micrograms X 29) as immunotherapy. PMID- 2987567 TI - [Determination of cerebral blood flow by the use of nitrous oxide and radioactive inert gases]. PMID- 2987569 TI - [Brain metabolism disorder associated with cerebral ischemia]. PMID- 2987568 TI - [Quantitative measurement of brain metabolism by positron CT]. PMID- 2987570 TI - [Agents to improve cerebrovascular circulation and cerebral metabolism--ATP and dilazep]. PMID- 2987571 TI - [Agents to improve cerebrovascular circulation and cerebral metabolism- vinpocetine]. PMID- 2987572 TI - [Viral gastroenteritis: serological characteristics and molecular epidemiology of human rotavirus]. PMID- 2987574 TI - [Diagnostic accuracy of CT, US and angiography in histologically verified hepatocellular carcinoma. Report on 125 foci in 84 cases]. PMID- 2987573 TI - [Mechanisms of antiherpes drugs and drug resistant virus--effect of BVDU on varicella zoster virus replication]. PMID- 2987576 TI - [Role of systemic chemotherapy in small cell lung cancer as part of a multidisciplinary treatment. I]. PMID- 2987575 TI - [CT detection of daughter nodules in hepatocellular carcinoma after lipiodol infusion via the hepatic artery]. PMID- 2987577 TI - [An epidemiological study on relationship between hepatitis B virus and hepatocellular carcinoma]. PMID- 2987578 TI - [Partial splenic embolization as a treatment for rupture of esophageal varices: a case report]. PMID- 2987579 TI - [A case of hepatocellular carcinoma with growth into the right atrium, pulmonary tumor embolism, and cerebral metastasis]. PMID- 2987580 TI - [A glucagon-secreting alpha-cell carcinoma of the pancreas]. PMID- 2987581 TI - [Clinical effects of local hyperthermia in patients with advanced hepatocellular carcinoma]. PMID- 2987583 TI - Hereditary hemolytic anemia with special reference to erythroenzymopathies. PMID- 2987584 TI - Alternating combination chemotherapy with a multimodality approach for small cell lung cancer. PMID- 2987582 TI - A case of hyperreninemic hypertension with unilateral hydronephrosis. AB - A patient with unilateral ureteral obstruction by urolithiasis at the ureteropelvic junction was accompanied by hypertension. Plasma renin activity (PRA) was high in peripheral veins and was significantly higher in the renal vein drained from the affected kidney than the contralateral. Infusion of angiotensin II antagonist or an oral administration of captopril, an angiotensin converting enzyme (ACE) inhibitor, resulted in a prompt drop of blood pressure. After pyelolithotomy was successfully performed, both the blood pressure and peripheral PRA completely normalized. It is suggested that the renin-angiotensin system might have played a major role in the mechanism of the accompanied hypertension. Hyperreninemia could have been caused by both renal ischemic vasoconstriction, which might be due to uretero-renal reflex, and increased synthesis of prostaglandins resulting from ureteropelvic obstruction. PMID- 2987586 TI - Comparative study of therapeutic effects of transcatheter arterial embolization and of intra-arterial one shot injection therapy against hepatocellular carcinoma. PMID- 2987585 TI - Clinical evaluation of arterial administration of SMANCS in oily contrast medium for liver cancer. PMID- 2987587 TI - Intratumoral injections of Nocardia rubra-cell wall skeleton for hilar type lung cancer in humans and Lewis lung carcinoma in mice. PMID- 2987588 TI - Differential effects of electroconvulsive shock on four models of experimentally induced aggression in rats. AB - The effects of electroconvulsive shock (ECS) on the hyperemotionality and muricide in olfactory bulbectomized rats (OB rat) were compared with those in spontaneous killer rats (SP rat), raphe lesioned rats (Raphe rat) and delta 9 tetrahydrocannabinol treated rats (THC rat). Single and chronic treatment of ECS inhibited the muricide and attack response to a rod, but did not affect the struggle response to handling and flight response to air blowing. Muricide was markedly inhibited by ECS in OB and SP rats, and was moderately inhibited in THC rats, while it was slightly inhibited in Raphe rats. The present result indicates that muricide of OB rats is a useful model for evaluating antidepressant drugs as this behavior is markedly inhibited by ECS in OB rats. Furthermore, it is suggested that the effect of ECS on muricide is different depending upon the methods to induce muricide, although muricide itself seems to be behaviorally similar. PMID- 2987589 TI - Effects of idebenone (CV-2619) on endogenous monoamine release and cyclic AMP formation in diencephalon slices from rats. AB - The addition of CV-2619 (10(-4) M) stimulated the release of endogenous 5 hydroxytryptamine (5-HT) markedly and that of norepinephrine (NE) slightly from slices of rat diencephalon, but it did not affect dopamine (DA) release. The addition of CV-2619 (10(-5)-10(-4) M) or 5-HT (10(-6)-10(-4) M) stimulated cyclic AMP formation in a concentration-dependent manner. Methysergide (10(-4) M), a 5 HT receptor blocker, almost completely blocked CV-2619 (10(-4) M)- and 5-HT (10( 4) M)-induced cyclic AMP formation. These results suggest that CV-2619 stimulates cyclic AMP formation via endogenous monoamine, particularly, 5-HT release. PMID- 2987590 TI - Selective interaction of DG-5128 with a low agonist affinity state of alpha-2 adrenoceptor. AB - Using rat cerebral cortex membranes, the inhibitory effect of DG-5128 against (3H)-clonidine binding was compared between low (alpha 2L) (in the presence of EDTA) and high (alpha 2H) affinity states (in the presence of excess magnesium) of alpha 2-adrenoceptor for agonists. The Ki value (pKi=6.79) of DG-5128 in the alpha 2L state was 6.4 times higher than the value in the alpha 2H state. Thus, DG-5128 produces alpha 2-adrenoceptor antagonism through the selective interaction with an alpha 2L state of the receptor. PMID- 2987591 TI - Stereoselective enhancement of nociception by opioids in different strains of mice. AB - The effects of Mr2096, Mr2097, diprenorphine, (-)bremazocine and Mr2266 on jumping latencies were evaluated in Swiss, CXBK, C57BL, CXBH and CBA strains of mice. Mr2096 and Mr2097 respectively produced analgesia and hyperalgesia in these strains of mice. Thus their effects on nociception were mediated by stereoselective opioid receptors. Diprenorphine also produced hyperalgesia which might be mediated by both mu and kappa receptors. Kappa receptors also appeared to mediate hyperalgesia at least in Swiss and CBA strains, as both (-)bremazocine and Mr2266 shortened the jumping latencies. PMID- 2987592 TI - Varicella encephalitis. PMID- 2987593 TI - Inhibition by retinoic acid of murine retrovirus-induced cellular transformation and tumor formation. AB - The effect of all-trans-retinoic acid (RA) on cellular transformation and on tumorigenicity of retrovirally transformed cells was investigated. RA treatment of NRK and NIH/3T3 cells transformed by BALB/c murine sarcoma virus (MuSV), Kirsten murine sarcoma virus (K-MuSV), and simian sarcoma virus resulted in a significant reduction in anchorage-dependent growth of only K-MuSV-transformed NRK cells. A 62% reduction in cell number was observed at 10(-5) M RA. In contrast, anchorage-independent growth induced by each of the viruses tested was suppressed by RA. Balb/cMSV3T3 cells showed the greatest level of sensitivity with a significant reduction in anchorage-independent growth occurring at 10(-9) M RA. The level of cytoplasmic retinoic acid-binding protein (CRABP) was determined in both parent and transformed cell lines. CRABP was present at a high level in all 3T3 cell types but was absent in all NRK cell lines. For testing the antineoplastic activity of RA in vivo, Balb/cMSV3T3 cells were injected intradermally into nude mice. Subsequent treatment of the tumor sites of these animals by topical application of RA resulted in a significant reduction in both tumor incidence and tumor size, confirming the in vitro results. Analysis of the level of v-onc mRNA revealed that inhibition of retroviral transformation by RA was not due to a decrease in transcription of the v-onc genes. PMID- 2987594 TI - Induction of tumors with cycasin in newborn and preweanling rats. AB - Comprehensive studies of carcinogenesis in newborn or preweanling SD rats were conducted under various dose schedules of cycasin (CAS: 14901-08-7) administration. When cycasin was given sc to newborn rats at day 0, tumors were detected in more than 80% of rats of both sexes; kidney tumors were by far the most common. The incidences of tumors declined in the older groups, namely, over 60% in both sexes in the 7-day group, 55% in males and 8.3% in females in 14-day rats, and 0% in 21-day groups. By multiple administration, tumor incidences elevated considerably. Administration ip of cycasin also gave rise to tumor induction in newborn rats. A total of 435 kidney tumors found in the experiments were studied pathologically. Most of them were classified as mesenchymal tumor; some of them metastasized. A few other tumors were found in the liver and colon. PMID- 2987596 TI - [Superoxide anion (O2-) release from mouse peritoneal macrophages associated with ingestion of mycobacteria and effects of O2- on intracellular multiplication of mycobacteria]. PMID- 2987595 TI - Inhibitory effect of dietary p-methoxybenzeneselenol on azoxymethane-induced colon and kidney carcinogenesis in female F344 rats. AB - The effect of dietary p-methoxybenzeneselenol on colon carcinogenesis induced by azoxymethane [(AOM) CAS: 25843-45-2] was studied in female F344 rats. Starting at 5 weeks of age, animals were fed the high-fat diet (control diet) or high-fat diet to which 50 ppm of p-methoxybenzeneselenol (experimental diet) had been added. At 7 weeks of age, all animals except the vehicle-treated controls were administered sc injections of AOM (15 mg/kg body wt, once weekly for 3 wk). Animals were fed the control and experimental diets until 1 week after carcinogen treatment when those animals receiving the p-methoxybenzeneselenol diet were fed the control diet until termination of the experiment. p-Methoxybenzeneselenol in the diet significantly inhibited the incidence (percentage of animals with tumors) of tumors in the colon and kidney, as well as the colon tumor multiplicity (adenomas and adenocarcinomas per animal). PMID- 2987597 TI - A solution to the enigma of atrophic alveolar ridges. PMID- 2987598 TI - [Multiple glomus tumors of the stomach]. PMID- 2987600 TI - [Late results of surgical treatment of patients with small-cell carcinoma of the lung]. PMID- 2987599 TI - [Glomus tumor of the peritoneal cavity associated with lymphogranulomatosis]. PMID- 2987601 TI - [Megestrol acetate in various doses in the treatment of metastatic breast carcinoma--clinical and endocrinologic studies]. AB - Both medroxyprogesterone acetate (MPA) and megestrol acetate (MA) are effective in the treatment of metastatic breast cancer. Although the dose-dependent mode of actions of MPA have been extensively clarified, there is still some uncertainty regarding the mode of actions and dosage of MA. Thirty-three patients with metastatic breast cancer were treated with various dosages of MA under a phase-II study. Eight patients were given 200 mg, 9 X 400 mg, 10 X 600 mg and 6 X 800 mg MA daily per os. The LH, FSH, TBI, T3, T4, TSH, ACTH, aldosterone, testosterone, prolactin and cortisol levels were determined regularly during treatment to enable the investigation of the pharmacodynamics of MA. A complete remission was achieved in two patients, a partial remission in seven patients and there was no change in eight patients (total responder rate 51.5%). The clinical and endocrine changes therefore suggest that the dose-dependent mode of actions of MPA and MA are identical. Equivalent dosages of MPA are 1000-1500 mg per os and of MA 160 200 mg. Furthermore, similar relationships between the endocrine changes and remission behaviour of MA and MPA have been observed. Persisting tumour remissions are inevitable under cortisol suppression and normal prolactin, aldosterone and ACTH levels. PMID- 2987602 TI - [Vinyl chloride induced hepatocellular carcinoma]. AB - On the occasion of a hitherto unique observation of three hepatocellular carcinomas in workers of the same industrial plant within 7 years following long term exposure to vinylchloride, the characteristics are discussed of a chemical carcinogenesis leading to two different malignant tumours: haemangiosarcoma and hepatocellular carcinoma. This carcinogenic sequence has been predicted by animal studies. It is not known why the transformation of hepatocytes into carcinoma is far rarer than of sinusoidal cells into sarcoma. Whereas the hepatocellular carcinoma predominantly develops in association with cirrhosis, vinyl chloride is able to cause cancer directly without other known co-carcinogenic agents. This hepatic carcinogenicity is dose-dependent. After the introduction of industrial prevention measures, a new initiation of the tumour is improbable. Nevertheless, because of its long latency period, estimated between 5 and 20 years, clinical manifestations are still possible. An early diagnosis by sonography and computertomography, possibly combined with puncture, in exposed persons or those formerly at high risk is conceivable, while laboratory data, even tumour markers are unreliable. Its fulminant course does not differ from that of other hepatocellular carcinomas and has until now hindered successful treatment. PMID- 2987603 TI - The pathophysiology of cholesterol metabolism in man. AB - The concentration of low density lipoprotein in human plasma depends on the balance between its rates of synthesis and catabolism. Although both processes appear to be independently regulated they occur side by side in the liver and may be linked via the activity of the high affinity low density lipoprotein receptor on hepatocyte membranes. Dietary changes such as cholesterol feeding or variation in fat content can promote synthesis of the lipoprotein without changing catabolism while other interventions (e.g. sequestrant resin therapy) have the opposite effect. These different responses may be explained on the basis of compartmentalisation of regulatory sterol pools in the liver cell. PMID- 2987604 TI - [AIDS and HTLV-III in West Germany: the status February 1985]. PMID- 2987606 TI - Relation of endogenous digoxin-like immunoreacting activities to salt balance and renal function in man. AB - We have previously shown that a natriuretic factor which is present in a small molecular weight fraction (IV) of serum and urine from salt loaded animals and healthy subjects, respectively, inhibits the Na-K-ATPase enzyme in vitro and also binds to a specific digoxin antibody. In the present study digoxin-like immunoreacting activity (DLIA) was therefore determined in the serum of healthy volunteers during low (35 nmol/day) and high (greater than 400 mmol/day) sodium intake and of patients with chronic renal failure and serum creatinine concentrations ranging from 127 to 757 mumol/l. DLIA was determined with a radioimmunoassay for digoxin in native serum and in the salt (III) and post-salt (IV) serum fractions eluted from a Sephadex G-25 column. DLIA in native serum of healthy subjects was less than 0.125 ng/ml. After gel filtration DLIA eluted exclusively in the small molecular weight salt (F III) and post-salt (F IV) fractions. Whereas DLIA increased in F III and decreased in F IV, total DLIA in F III + IV slightly increased from 0.37 +/- 0.03 to 0.49 +/- 0.05 ng/ml (p less than 0.01) with the change from low to high sodium intake. DLIA in native serum of uremic patients ranged from 0 to 1.70 ng/ml and was detectable consistently only in patients with serum creatinine concentrations above 250 mumol/l. DLIA in F III which averaged 0.22 +/- 0.04 ng/ml and total activity which ranged from 0.11 to 0.88 ng/ml closely correlated with the degree of renal impairment (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987605 TI - Cytomegalovirus DNA in colorectal carcinoma tissues. AB - Numerous studies had linked human cytomegalovirus (HCMV) infections with neoplasia. Among various other malignant tumors, colonic carcinoma tissues were reported to contain DNA sequences hybridizing with DNA extracted from virus particles. Gene technology allowed us to use a cloned viral DNA library to measure HCMV in colorectal tumors more specifically. Four of 38 tissue specimens did contain DNA sequences homologous to cloned viral DNA probes; however, in each of those cases, identical hybridization patterns were seen with specimens from non-infiltrated surrounding intestinal wall. The amount of HCMV DNA in normal tissues was at least as much as in tumor biopsies. We conclude that nucleic acid hybridizations at high sensitivity with moleculary cloned HCMV DNA did not support the notion of a correlation between colorectal carcinomas and human cytomegalovirus. PMID- 2987607 TI - Raised sodium pump activity and a circulating sodium transport inhibitor demonstrated on red blood cells of patients with untreated essential hypertension: correlation of pump activity with potassium permeability. AB - We have studied sodium potassium ATPase activity, the effect of endogenous plasma on sodium pump activity, potassium permeability and intracellular sodium and potassium concentrations in normotensive subjects without (n = 36) and with (n = 33) a positive family history of hypertension, and in patients with untreated essential hypertension (n = 52). Sodium pump activity was studied as ouabain sensitive uptake of rubidium 86 in washed red blood cells, incubated in an artificial medium closely resembling the anorganic constituents of plasma. Any influence of endogenous plasma on sodium pump activity was investigated by re incubating the washed red blood cells in their own plasma and comparing ouabain sensitive rubidium uptake in the two media. To correct for any possible differences in external potassium concentration, a function for the relation between extracellular potassium concentration and absolute transport rates was derived experimentally. From this, actual transport rates in plasma were corrected by computer to an extracellular potassium concentration of 4.0 mmol/l. Sodium pump activity, concentration of circulating sodium transport inhibitor, potassium permeability and intracellular electrolytes were not statistically different in subjects with and without a positive family history of hypertension. Hypertensives had significantly raised sodium pump activity in artificial medium, but not when red cells were re-incubated in their own plasma. Thus, endogenous plasma inhibited the sodium pump by between 12% and 15%. Hypertensives also had a significantly raised potassium permeability. Potassium permeability and sodium pump activity were correlated significantly. Intracellular sodium concentrations were similar in normotensives and hypertensives, but the later showed a significantly lower intracellular potassium concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987608 TI - Mechanism of diuretic-induced hypopotassemia in human hypertension. AB - Diuretic treatment (hydrochlorothiazide) induced a marked decrease of red cell membrane Na+K+ ATPase activity in excessive potassium loser hypertensive patients. The decreased activity occurred within 2-4 weeks of treatment and returned to baseline in 4-6 weeks after cessation of treatment. Simultaneously, red cell sodium increased, potassium decreased together with increased 24-h urinary excretion. The persistent low serum potassium may be due to impaired absorption of potassium from the gut as a result of suppressed enzyme activity since total body potassium appears also to decrease. The decreased Na+K+ ATPase activity may be due to a direct effect of the diuretic on cell membrane. PMID- 2987609 TI - Vasoconstrictor effects of ouabain: mechanisms of action. PMID- 2987610 TI - [Glomus tumor of the small intestine with metastasis to the liver]. PMID- 2987611 TI - [Problems in follow-up care of adolescents formerly treated by ward personnel during hospitalization in the child and adolescent psychiatry department]. PMID- 2987612 TI - Experimental Herpes neonatalis in SA 8-infected baboons (Papio cynocephalus). AB - 13 newborn baboons were experimentally infected with a baboon-derived herpesvirus SA 8-strain. Intratracheal infection produced herpetic tracheobronchopneumonia, intravenous inoculation resulted in adrenal necrosis similar to that in human neonatal herpes infection. PMID- 2987613 TI - Epizootiological observations of natural and experimental infection with sialodacryoadenitis virus in rats. AB - The epizootiology of sialodacryoadenitis (SDA) was studied in experimentally and naturally infected rats. The infectivity of SDA virus (SDAV) in intranasally infected rats was lost by seven days after infection as determined by contact transmission. After experimental infection, SN antibody appeared earlier and titers were detectable longer than CF antibody. The prevalence of SN antibody positive rats in naturally infected colonies remained high, whereas an increase in the prevalence of CF antibody-positive rats appeared to coincide with the introduction or resurgence of SDAV. A SDAV-free colony was established by allowing recovered dams to litter in a separate room. A spontaneous cessation of SDAV infection also was observed in an enzootically-infected colony. Clinical observations indicated that SDA can occur as a mild or asymptomatic disease, and that its clinical expression may vary from one inbred strain to another. PMID- 2987614 TI - Properties and compartmentalization of the testicular receptor for 1,25 dihydroxyvitamin D3. AB - Adult rat testis contains a specific, high-affinity, low-capacity binding protein for 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) with properties similar to 1,25 (OH)2D3 receptors in other tissues. The receptor sediments at 3.5 +/- 0.2 S20,w in high-salt sucrose density gradients, but aggregates in low-salt gradients. Binding of 1,25-(OH)2D3 was abolished by trypsin, but not by DNase or RNase. Binding was also heavily reduced by the sulfhydryl alkylating agent, N ethylmaleimide, and by the mercurial reagent, mersalyl, showing that free, reduced SH-groups are necessary for hormone-binding activity. The receptor shows high affinity for 1,25-(OH)2D3 (Kd = 3 X 10(-11) M), but low capacity (Nmax = 8 fmol/mg protein) and is specific for 1,25-(OH)2D3 (Affinity: 1,25-(OH)2D3 greater than 1,24(R),25-(OH)3D3 greater than 25-OH-D3 greater than 1 alpha-OH-D3 greater than 24(R),25-(OH)2D3 much greater than 17 beta-estradiol, testosterone, dexamethasone, R5020, progesterone). With 0.6 nM [3H]1,25-(OH)2D3 and at 0 degrees C, maximum specific binding was achieved after 4 h, and the occupied receptors were stable for more than 24 h. The dissociation of hormone-receptor complexes was temperature-dependent and very slow at low temperature (t1/2 (0 degrees C) much greater than 48 h). At 0 degrees C, the second order association rate constant and the pseudo-first order dissociation rate constant were 2.7 X 10(7) M-1 min-1 and 2 X 10(-5) min-1, respectively. Receptors for 1,25-(OH)2D3 are present in similar amounts in isolated seminiferous tubules and interstitial tissue of adult rats. No specific binding of [3H]1,25-(OH)2D3 could be detected in cultured immature Sertoli cells, cultured immature peritubular (myoid) cells or crude germ cells. PMID- 2987615 TI - Synthesis and biological activity of 3 beta-fluorovitamin D3: comparison of the biological activity of 3 beta-fluorovitamin D3 and 3-deoxyvitamin D3. AB - The alteration in the biologic activity of the vitamin D3 molecule resulting from the replacement of a hydrogen atom with a fluorine atom is a subject of fundamental interest. To investigate this problem we synthesized 3 beta fluorovitamin D3 6 and its hydrogen analog, 3-deoxyvitamin D3 7, and tested the biologic activity of each by in vitro and in vivo methods. Contrary to previous reports which showed that 3 beta-fluorovitamin D3 was as active as vitamin D3 in vivo, we found that the fluoro-analog was less active than vitamin D3. With regard to stimulation of intestinal calcium transport and bone calcium mobilization in the D-deficient hypocalcemic rat, 3 beta-fluorovitamin D3 showed significantly greater biologic activity than its hydrogen analog, 3-deoxyvitamin D3. In the organ-cultured, embryonic chick duodenum, 3 beta-fluorovitamin D3 was approx 1/1000th as active as the native hormone, 1,25-dihydroxyvitamin D3, while 3-deoxyvitamin D3 was inactive even at microM concentrations, in the induction of the vitamin D-dependent, calcium-binding protein. With regard to in vitro activity in displacing radiolabeled 25-hydroxyvitamin D3 from vitamin D binding protein and radiolabelled 1,25-dihydroxyvitamin D3 from a chick intestinal cytosol receptor, 3 beta-fluorovitamin D3 and 3 beta-deoxyvitamin D3 both showed very poor binding efficiencies when compared with vitamin D3. Our results show that the substitution of a fluorine atom for a hydrogen atom at the C-3 position of the vitamin D3 molecule results in a fluorovitamin 6 with significantly more biological activity than its hydrogen analog, 3-deoxyvitamin D3 7. PMID- 2987617 TI - The catalytic mechanism of serine proteases II: The effect of the protein environment in the alpha-chymotrypsin proton relay system. AB - The proton relay system of alpha-chymotrypsin is analyzed by the INDOISCRF method. The effects of the protein electric and polarization fields are explicitly introduced in the calculations. It is shown that the multicharged structure Ser-His+Asp- is the most sensitive, from an energetic view-point, towards the protein surrounding effects. Variations in the permanent and polarization fields are discussed, as well as the influence of the substrate and one water molecule localized in the active site of the enzyme. The catalytic role of such changes is conjectured. PMID- 2987616 TI - Relationships between unconjugated and sulphated steroids in porcine primary Leydig cell culture. AB - Steroidogenesis in immature porcine Leydig cells was investigated in primary culture at 48-84 h under basal conditions and in the presence of hCG. The basal accumulation of unconjugated steroids was close to linear only during the first 4 h of study, whereas the sulphate-conjugated steroids accumulated essentially linearly over the 36 h experimental period. At the last time point, 95% of the steroids measured were sulphated. Stimulation with hCG (1 ng/ml) led to a still more pronounced sulphate conjugation, and approx 99% of the steroids measured were sulphated at 36 h. Under maximal stimulation with hCG (100 ng/ml) the sulphates accounted for 74% of the total steroids measured at 36 h. Testosterone, androstenedione, dehydroepiandrosterone, 5-androstene-3 beta, 17 beta-diol and estrone were usually quantitatively the most important unconjugated steroids, and sulphated dehydroepiandrosterone, estrone, testosterone and 5-androstene-3 beta, 17 beta-diol were the most important steroid sulphates, especially following maximal stimulation of the cultures. These data emphasize the importance of steroid sulphates in porcine testicular steroid metabolism. Under stimulation with hCG, there was a rapid response in testicular steroidogenesis, initially seen as a rapid increase in the secretion of unconjugated and sulphated steroids. At approx 4-12 h, the rate of sulphate conjugation appeared to reach or even to exceed that of steroid biosynthesis, which lead to stabilisation or a decrease in the concentrations of unconjugated steroids. Only high doses of hCG, 10-100 ng/ml, were then able to lead to a net accumulation of unconjugated steroids, at 24-36 h of incubation with hCG. PMID- 2987618 TI - Mediastinal germ cell tumors in childhood. A clinical and pathological study of 21 cases. AB - Twenty-one patients aged 16 years or less had been treated for a primary mediastinal germ cell tumor at the Children's Hospital, Boston Massachusetts, during the last 54 years. There were 13 boys and eight girls with the average age at diagnosis being 7 years (range 2 weeks to 16 years). Twelve mediastinal germ cell tumors were classified as pure teratoma, five contained embryonal carcinoma admixed with other germ cell components, and four were pure embryonal carcinoma. Of 12 patients with pure teratoma, 10 underwent complete surgical resection and were alive and well 1 to 13 years later; two children left untreated died of complications related to local tumor growth. Complete surgical resection was possible for only two of nine patients with embryonal carcinoma; both received adjuvant therapy and were alive and well 3 and 20 years later. Seven patients received radiation and/or chemotherapy but died of residual or metastatic disease. Successful treatment for children with embryonal carcinoma requires an operation aimed at either debulking or complete resection (if possible) coupled with early and aggressive combination chemotherapy. The role of radiation in primary therapy remains undefined with regard to curative intent. PMID- 2987619 TI - Chest wall invasion in carcinoma of the lung. Therapeutic and prognostic implications. AB - From 1974 through 1983, 125 patients underwent operation at Memorial Sloan Kettering Cancer Center for non-small cell carcinoma of the lung invading the chest wall. (Excluded are those with superior sulcus tumors or distant metastases at presentation.) Eighty patients were male and 45 were female. Ages ranged from 33 to 88 years (median 60 years). Histologically, the tumors were epidermoid carcinoma in 46%, adenocarcinoma in 46%, and large cell carcinoma in 8%. All patients underwent thoracotomy (pneumonectomy 19, bilobectomy seven, lobectomy 75, wedge resection 10, and no pulmonary resection 14), with an operative mortality of 4%. At thoracotomy, mediastinal lymph node dissection was routinely performed, and the postsurgical stage was T3 N0 M0 in 53%, T3 N1 M0 in 13%, and T3 N2 M0 in 34%. Extrapleural resection was performed in 66 patients. En bloc resection of chest wall and lung was performed in 45 patients with an operative mortality of 2%. Complete resection of tumor was possible in 77 patients (62%). Extension of tumor beyond the parietal pleura significantly decreased resectability. The median survival of 48 patients having incomplete resection was 9 months, despite perioperative interstitial and external radiation. The actuarial 5 year survival rate (Kaplan-Meier) of 77 patients having complete resection was 40%. This percentage was not significantly influenced by the patient's age or sex or by tumor size or histologic type. Lymphatic metastases significantly reduced survival, with a 5 year actuarial survival rate of 56% for patients with T3 N0 M0 disease and 21% for those with T3 N1 M0 or T3 N2 M0 disease (p = 0.005). The extent of tumor invasion of the chest wall appeared to influence survival, but in the absence of lymphatic metastases the difference at 5 years was not significant. Complete resection offers a significant chance for long-term survival in lung cancer directly extending into parietal pleura and chest wall. Extrapleural resection or en bloc chest wall resection can be performed with a low operative mortality and an expected 5 year survival in excess of 50% in the absence of lymphatic metastases. PMID- 2987620 TI - Decline of natural cytotoxicity of human lymphocytes following infection with human T-cell leukemia/lymphoma virus (HTLV). AB - Cell-mediated natural cytotoxicity (CMNC) of fresh or long-term cultured lymphocytes collected from HTLV-positive patients or infected in vitro with the virus, was tested against K562 target cells. Severe depression of reactivity was found in fresh lymphocytes of three patients with advanced disease, in 12 in vitro established T-cell malignant lines, and two HTLV-infected cord blood (C5/MJ and C91/PL) lines. Moreover, all (eight) HTLV-1 infected cell lines listed showed a significant inhibition of CMNC of peripheral blood lymphocytes of healthy donors. Whether virus infection promotes the outgrowth of pre-existing suppressor cells and/or produce changes of the T-lymphocyte function is unknown. PMID- 2987621 TI - Responsiveness of the in vitro hematopoietic microenvironment to platelet-derived growth factor. AB - The adherent or stromal cells in human long-term marrow cultures, a possible in vitro counterpart of the in vivo microenvironment, were investigated for responsiveness to platelet-derived growth factor (PDGF). Many stromal cells from cultures derived from normal donors as well as from patients with chronic myelogenous leukemia, bore receptors to PDGF and were stimulated to incorporate [3H]-thymidine by highly purified PDGF and to a lesser extent by epidermal growth factor (EGF). These data suggest that PDGF and perhaps EGF may be involved in the regulation of the in vitro microenvironment and, therefore, of normal and possibly neoplastic hematopoiesis. PMID- 2987623 TI - The monoclonal antibodies to p19 of HTLV should not be used alone to screen for the virus in T-cells. Comment. PMID- 2987622 TI - The effect of the leukemic cell line HL60 and acute myeloblastic leukemic cells before and after induction of differentiation on normal pluripotent hematopoietic progenitors (CFU-GEMM). AB - A leukemic cell line (HL60) and acute myeloblastic leukemia (AML) cells from six patients were co-cultured with normal marrow cells to assess their effects on growth of normal CFU-GEMM. The effects of the following inducers: 12-0 tetradecanoyl-phorbol-13-acetate (TPA), retinoic acid (RA), dimethylsulphoxide (DMSO), 1-25 (OH) D3 (Vitamin D3) and PHA-LCM on both the HL60 and AML cells, were studied. Inhibition of growth of normal CFU-GEMM was observed in the co cultures in the presence of 1 X 10(4) HL60 or AML leukemic cells/ml. This inhibition was reversed by pretreating the HL60 line with vitamin D3, TPA and RA. No effect on growth of CFU-GEMM was noted when DMSO and PHA-LCM were used. AML cells were morphologically induced to differentiate by TPA or RA in all six cases. In three cases, reversal of inhibition of growth of normal pluripotent hemopoietic progenitors occurred and in three the inhibition of growth persisted. Regulation of inhibition by different inducers did not seem to correlate in all cases with morphological differentiation. PMID- 2987624 TI - IL-2 production and response in vitro by the leukocytes of patients with acquired immune deficiency syndrome. AB - In order to investigate the nature of the T cell defect associated with the acquired immune deficiency syndrome (AIDS) we studied the ability of peripheral blood mononuclear cells from 8 patients with Kaposi's sarcoma (KS), 2 with opportunistic infection (OI), 23 with AIDS-related symptoms complex (ARC) without KS or OI (ARC), and 29 heterosexual controls to produce interleukin II (IL-2) on phytohemagglutinin (PHA) stimulation and to respond to exogenously supplied IL-2. Patients with AIDS as well as those with ARC produced adequate levels of IL-2 in response to lectin stimulation when compared to controls (AIDS, 3.07 +/- 1.98 units; ARC, 3.03 +/- 1.89 units; controls, 3.75 +/- 1.52 units). However, the ability of these patients' cells to respond in vitro to exogenously supplied IL-2 as measured on short-term PHA-stimulated T cell blasts, was found to be severely impaired in patients with AIDS and ARC (AIDS, 22.4 +/- 6.0 X 10(-3) cpm; ARC, 20.1 +/- 4.2 X 10(-3) cpm; control, 41.4 +/- 4.2 X 10(-3) cpm). This impairment was associated with diminished expression of the IL-2 receptor on 7-day-old lectin-stimulated T cells from both patient groups (AIDS, 17.7 +/- 5.7; ARC, 36.8 +/- 4.4; control 71.8 +/- 1.7). These results should be considered when IL-2 is proposed as potential therapy in the treatment of AIDS. They also suggest that the nature of the AIDS defect is related to impaired hormone receptor expression. PMID- 2987625 TI - Chemical identification of a prostaglandin-induced T suppressor (PITS). AB - Prostaglandin E1 is known to block the activation and function of immunocompetent cells. Previous studies from this laboratory have established that one pathway where prostaglandin E1 acts involves the stimulation of a glass-wool adherent T suppressor cell. Prostaglandin stimulates this suppressor cell to release a suppressor lymphokine(s), termed the prostaglandin-induced T cell-derived suppressor (PITS). We report here the further characterization of this factor using high-pressure liquid chromatographic analyses, precursor labeling studies, and bioanalysis, all of which indicate that this factor contains a leukotriene. PMID- 2987627 TI - A role for copper in mediating oxidative damage associated with degenerative disease processes seems to be more imaginary than real. PMID- 2987626 TI - Interleukin 2 stimulation of protein kinase C membrane association: evidence for IL-2 receptor phosphorylation. PMID- 2987628 TI - Copper can both mediate and prevent oxidative damage. PMID- 2987629 TI - [A 68 year-old woman with cardiac insufficiency, muscular weakness and lumbar pain]. PMID- 2987630 TI - [The ideal blood donor]. PMID- 2987631 TI - [Cytomegalovirus and congenital infection--serologic pregnancy screening is not suitable]. PMID- 2987632 TI - [HTLV-III infections and AIDS--current epidemiology and serology]. PMID- 2987633 TI - [LAV/HTLV-III--new discoveries on its properties, isolation, vaccination and therapy]. PMID- 2987634 TI - Dissociation between adrenocorticotropin and corticosterone responses to restraint after previous chronic exposure to stress. AB - The effect on emotional reactivity produced by a model for chronic stress in which different types of acute stresses were randomly combined for 29 days was studied in adult male rats. Chronically stressed rats showed a slight decrease in body weight gain and an increase in relative adrenal weight. Chronic stress did not modify defecation rate but reduced exploratory activity in the holeboard. Neither basal nor acute-stress induced levels of adrenocorticotropin (ACTH) were modified by previous chronic stress. Likewise, basal corticosterone levels were similar in both groups. However, corticosterone response to acute restraint stress was higher in chronically stressed than in control rats. The mechanisms underlying the dissociation between ACTH and corticosterone as well as its possible implications are discussed. PMID- 2987635 TI - Prenatal stress impairs maternal behavior in a conflict situation and reduces hippocampal benzodiazepine receptors. AB - Maternal behavior (pup retrieval) was assessed in prenatally stressed rats during control and conflict situations (having to pass through an airstream) when their pups were 4-5 days old. There was no difference in pup retrieval between experimental and control rats under normal conditions but only 52% of the former retrieved their pups during the conflict situation, compared with 96% of the controls. Catecholamine (CA) levels in the arcuate nucleus (Arc.n.) and noradrenaline in the medial preoptic nucleus (POM) were not altered in prenatally stressed females, but their dopamine levels in the POM tended to be lower (p less than 0.1). The number of benzodiazepine (BZ) receptors in the hippocampi of prenatally stressed females was significantly lower than in controls. We conclude from these results that random prenatal noise and light stress increases the vulnerability to stressful situations in the female offspring during adulthood, which may be accompanied by altered CA function in the hypothalamus and BZ binding in the hippocampus. PMID- 2987636 TI - An ACTH 4-9 analog impairs selective attention in man. AB - Male adults were tested in a dichotic listening task, providing electrophysiological measures of selective attention. Subjects were tested twice, 60 min after oral administration of either 40 mg of ACTH 4-9 analog, or placebo. Averaged auditory evoked potentials (AEPs) to tone pips when attended and when unattended, EEG spectra, heart rate and blood pressure, and behavioral performance were measured during task performance. ACTH 4-9 analog treatment impaired selective attention as indicated a) by a diminished difference between evoked potential waveforms to attended and to unattended tone pips, b) by an impaired behavioral signal detection performance. Furthermore, frontal EEG theta activity slowed down after ACTH 4-9 analog. With time on task, however, there was no decay, but an improvement of selective attention after peptide administration. Differences in attention could not be due to concurrent changes in general cortical and autonomic arousal as indicated by EEG alpha activity, blood pressure and heart rate. Since separate analyses of the AEPs revealed an increased processing of the unattended tone pips in the ACTH 4-9 analog sessions the impaired selective attention under ACTH 4-9 analog may be described as an inability to suppress processing of irrelevant or distracting stimuli. PMID- 2987637 TI - The effects of various cognition-enhancing drugs on in vitro rat hippocampal synaptosomal sodium dependent high affinity choline uptake. AB - The purpose of the present study was to compare the effect of seven drugs, that have been reported to enhance cognitive functions, on rat hippocampal cholinergic neuronal activity. The latter was assessed by measuring the effects of the drugs on in vitro sodium-dependent high affinity choline uptake (HACU) into rat hippocampal synaptosomes 30 minutes after their in vivo administration. 3,4 Diaminopyridine (0.1 mg/kg IP), like pramiracetam (44 and 88 mg/kg IP), increased HACU with higher or lower doses being ineffective. Centrophenoxine (100 mg/kg IP) decreased HACU. Piracetam (100 and 500 mg/kg IP), aniracetam (10-200 mg/kg PO), lysine vasopressin (0.005-0.05 mg/kg IM) and 4-aminopyridine (0.01-3.0 mg/kg IP) were ineffective. The results indicate that 3,4-diaminopyridine and centrophenoxine, like pramiracetam may be increasing cognitive function in part by affecting hippocampal cholinergic neuronal activity. In addition, the findings indicate the usefulness of using in vitro HACU as a biochemical measurement to assess the potential effect of cognitive-enhancing drugs on cholinergic neuronal activity in vivo. PMID- 2987638 TI - Mu1: a very high affinity subtype of enkephalin binding sites in rat brain. AB - Displacement studies of [3H]-[D-Ala2-MePhe4-Gly-ol5]-enkephalin ([3H]-DAGO) and [3H]-[D-Ala2-D-Leu5]-enkephalin ([3H]-DADL) by the corresponding unlabeled ligands show that there are at least three classes of sites which bind these enkephalin analogs with high affinity. Using computer modeling, the introduction of the third site significantly improved the goodness of fit in ten consecutive experiments. These sites appear to correspond to the mu, delta and mu 1 sites, with mean dissociation constants of 11, 1.3 and 0.9 nM for DADL and 2.5, 300 and 0.3 nM for DAGO, respectively. PMID- 2987639 TI - Partial chemical characterization of cyclopyrrolones ([3H] suriclone) and benzodiazepines ([3H]flunitrazepam) binding site: differences. AB - Rat hippocampus membranes were treated with several protein modifying reagents (iodoacetamide, N-ethylmaleimide, tetranitromethane and N-acetylimidazole). The effects of these treatments on the binding sites of cyclopyrrolones ([3H] suriclone), a new chemical family of minor tranquilizers, and benzodiazepines ([3H] flunitrazepam) were investigated. Here we show that both ligands are similarly sensitive to cysteine alkylation: [3H] suriclone and [3H] flunitrazepam binding are reduced by iodoacetamide and slightly increased by N-ethylmaleimide. On the contrary they are clearly differenciated by tyrosine modification: [3H] suriclone binding is not changed whereas [3H] flunitrazepam binding is increased by tetranitromethane and decreased by N-acetylimidazole. Our present findings and published evidence suggest cyclopyrrolones and benzodiazepines bind to distinct sites or to different allosteric forms of the benzodiazepine receptor. PMID- 2987640 TI - Initial clinical evaluation of gadolinium DTPA for contrast-enhanced magnetic resonance imaging. AB - Gadolinium DTPA was evaluated as an intravenous contrast agent for magnetic resonance imaging in 15 patients with primary or secondary intracranial neoplastic disease. T1 and T2 weighted images were obtained prior to contrast administration. T1 weighted spin echo 35/800 (TE/TR) images were utilized to detect enhancement. The increase in signal intensity observed, identifying areas of breakdown of the blood-brain barrier (BBB), was similar in magnitude to the contrast enhancement observed on CT. This permitted differentiation of neoplastic tissue from surrounding cerebral edema on MRI. Direct visualization of otherwise "isomagnetic" lesions was also demonstrated. The use of intravenous contrast media should significantly extend the diagnostic potential and specificity of magnetic resonance imaging. PMID- 2987641 TI - Do cyclic AMP concentrations in saliva reflect PTH biologic activity? AB - The aim of this study was to investigate whether the concentration of cAMP in saliva might reflect the biologic activity of parathyroid hormone (PTH), as nephrogenous levels of cAMP reflect parathyroid function. Patients with primary hyperparathyroidism (HPT) with elevated levels of PTH, nephrogenous cAMP, and serum calcium were found to have cAMP levels not significantly different from control subjects. When the levels of these parameters were reduced following parathyroidectomy, salivary cAMP concentrations were not affected. Similarly, an oral calcium loading test (which decreased both PTH and nephrogenous cAMP in normal subjects) failed to influence salivary cAMP levels. Basal salivary cAMP concentrations were low in secondary hyperparathyroid patients due to chronic renal failure and rose almost fivefold following dialysis despite the stable plasma cAMP levels and salivary flow rate. We conclude that salivary cAMP concentrations do not appear to be influenced by changes in circulating biologically active PTH. However, salivary cAMP concentrations do change in response to certain perturbations; this suggests that they are regulated by some other modulator(s). PMID- 2987642 TI - Excision of copia element in a revertant of the white-apricot mutation of Drosophila melanogaster leaves behind one long-terminal repeat. AB - The spontaneous mutation white-apricot (wa) in Drosophila melanogaster has a considerably lighter eye colour than the wild-type, and is caused by the insertion of a copia transposable element into a small intron of the white gene. We have analyzed an X-ray induced wa revertant (waR59K1), whose eye pigmentation is incompletely restored, by in situ hybridization, Southern blotting and sequencing analysis. At the site where copia had originally inserted, we found one long terminal repeat of copia, flanked by a 5 bp duplication with the same polarity as the direction of transcription of the white locus. These results suggest that the wa reversion is due to homologous recombination between the two long terminal repeats of copia. PMID- 2987643 TI - Localization of the upstream regulatory sites of yeast iso2-cytochrome c gene. AB - In order to study the regulation of expression of the iso2-cytochrome c gene, we have constructed a fused gene between the 5'flanking region of the gene coding for the yeast iso2-cytochrome c and the coding region of the E. coli beta galactosidase lacZ gene. When introduced in yeast cells this hybrid gene is expressed and regulated like the production of iso2-cytochrome c: it is under the control of the general catabolic repression and of the unlinked trans-acting CYP1 gene whose CYP1-18 allele causes an overproduction of iso2-cytochrome c. The expression of hybrid genes whose upstream region has been progressively shortened or altered by internal deletions was studied either in wild-type CYP1+ cells or in cells carrying the CYP1-18 allele grown either on glucose or on glycerol. It appears that the expression and the regulation of the iso2-cytochrome c gene is controlled by an upstream regulatory site composed of a positive and a negative element. This site is the target of regulation by the CYP1 gene product and, directly or through this gene, of the control by the general catabolic repression. PMID- 2987644 TI - Physical characterization of the genome of the Myxococcus xanthus bacteriophage MX-8. AB - We have constructed a restriction map for the genome of bacteriophage MX-8 from Myxococcus xanthus using the enzymes PvuII, MboI, and EcoRI. The phage genome size, as determined by restriction analysis, is 51.7 +/- 0.6 Kb. Double digestions, redigestions of isolated fragments, and crossed-contact hybridization of partial digestion products show that the restriction map is circular. Restriction analysis and Southern hybridization show that the phage DNA molecules are packaged sequentially from a concatemer starting from a specific site which we have mapped. The DNA molecules have an average terminal redundancy of approximately 8% and are circularly permuted over at least 40% of the genome. PMID- 2987645 TI - The Klebsiella aerogenes glutamate dehydrogenase (gdhA) gene: cloning, high-level expression and hybrid enzyme formation in Escherichia coli. AB - The NADP-dependent glutamate dehydrogenase gene of Klebsiella aerogenes was cloned in E. coli in the expression plasmid pRK9. The cloned gene shows a high level of expression in E. coli in the hybrid plasmid pKG3 and such expression is independent of the vector promoter, as shown by experiments in which the promoter was deleted. Active hybrid GDH hexamers were shown in cell-free extracts of an E. coli strain carrying cloned gdhA genes of both E. coli and K. aerogenes. The nucleotide sequence of the N-terminal coding region of the K. aerogenes gdhA gene was determined and found to be strongly homologous with that of E. coli. PMID- 2987646 TI - Homologous recombination of polyoma virus DNA in mouse cells. AB - We have produced nonviable deletion mutants of polyoma virus in order to study homologous recombination after DNA transfection into mouse cells. The frequency of recombination was determined by the formation of infectious virus. It was dependent on the amount of DNA transfected and the size of the region of homology between the mutations. Recombination frequencies were highest when both mutated genomes were transfected in closed circular form rather than after linearization of one or both of the recombination partners. The system described may be useful for a more detailed analysis of physiological and genetic conditions influencing the frequency of homologous recombination in mouse cells as well as to study enzymes involved and intermediates produced in this process. PMID- 2987647 TI - Cloning and characterization of the rDNA repeat unit of Podospora anserina. AB - DNA coding for ribosomal RNA in Podospora anserina has been cloned and was found as a tandemly repeated 8.3 kb sequence. The cloned rDNA was characterized by restriction endonuclease mapping. The location of 5.8S, 18S and 28S rRNA coding regions was established by DNA-RNA hybridization and S1 nuclease mapping. The organization of P. anserina rRNA genes is similar to that of Neurospora crassa and Aspergillus nidulans. The rDNA unit does not contain the sequence coding for 5S RNA. PMID- 2987648 TI - Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces. AB - Promoter-probe plasmid vectors were used to isolate putative promoter-containing DNA fragments of three Streptomyces antibiotic resistance genes, the rRNA methylase (tsr) gene of S. azureus, the aminoglycoside phosphotransferase (aph) gene of S. fradiae, and the viomycin phosphotransferase (vph) gene of S. vinaceus. DNA sequence analysis was carried out for all three of the fragments and for the protein-coding regions of the tsr and vph genes. No sequences resembling typical E. coli promoters or Bacillus vegetatively-expressed promoters were identified. Furthermore, none of the three DNA fragments found to be transcriptionally active in Streptomyces could initiate transcription when introduced into E. coli. An extremely biased codon usage pattern that reflects the high G + C composition of Streptomyces DNA was observed for the protein coding regions of the tsr and vph genes, and of the previously sequenced aph gene. This pattern enabled delineation of the protein-coding region and identification of the coding strand of the genes. PMID- 2987649 TI - Effect of concentration on the subsequent fate of plasmid DNA in human fibroblasts. AB - The physical fate of plasmid DNA after entry into human fibroblasts was studied using Southern hybridisation and electron microscopy. Exposure of the cells (5 X 10(5) per well) to pC194 DNA-CaPi, containing 50 micrograms plasmid DNA, resulted in the occasional formation of interlocked molecules. Exposure to a co precipitate containing 100 micrograms pC194 plasmid DNA per well resulted in an increase of interlocked molecules by a factor of 10-20 relative to the number of monomers. In addition, new classes of molecules were observed. After prolonged incubation of the cells exposed to the higher DNA concentration, the plasmid DNA was partly contained in structures with a very low electrophoretic mobility. Upon restriction endonuclease digestion of the re-extracted DNA, a pattern of bands was observed, suggesting the involvement of illegitimate recombination between non-random plasmid DNA sequences in the formation of the new classes of molecules. PMID- 2987650 TI - Congenital neoplasia--a stem cell pathology. AB - The theory presented herewith provides a unified explanation for neoplasia and teratogenesis, both regarded here as stem cell disorders. Proliferating cells in the organism form two mutually exclusive classes, immortal stem cells and transitional cells whose life span is limited. Any lasting change in the organism e.g. tissue differentiation or neoplasia, is determined by stem cells. Congenital as well as adult neoplasms exhibit three key features: maturation arrest, stem cell pool expansion and increased cell turnover and they progress through the following states: dysplasia, neoplasia in situ, benign stage (e.g. polyp) and overt neoplasia. The neoplasm is regarded here as an organ with a purpose intended to supply the organism with a missing substance. Besides serving as tissue progenitors, stem cells are postulated here to secrete a substance 'A' necessary for a proper tissue function. Carcinogens interfere with the substance production mainly by depleting stem cells so that less 'A' is produced. The capacity of the adult to replenish the depleted stem cells is limited, and the missing substance has to be formed by an alternative way i.e. by Neoplasia. The 'A' substitute formed by the neoplasm denominated as 'B' is however less efficient than 'A'. Neoplastic growth thus depends upon the relative abundance of substances 'A' and 'B'. Since in the growing organism, stem cells multiply, some of the missing 'A' is replenished by them and the neoplasm may regress. PMID- 2987651 TI - [ACE inhibitors: a new pharmacological principle]. PMID- 2987652 TI - [Gyrase inhibitors]. PMID- 2987653 TI - Influence of maternal immunity on the outcome of murine coronavirus JHM infection in suckling mice. AB - Adult C3H mice are resistant to intraperitoneal infection with murine coronavirus JHM, whereas suckling offspring of non-immune females are susceptible. Resistance can be conferred on suckling C3H mice by postnatal transmission of maternal immunity, if transfer precedes infection. Suckling mice succumb to infection even when they receive maternal antibodies within 1 day after infection. Prenatal transmission alone without subsequent postnatal transmission of maternal immunity is not sufficient to provide resistance. Persistence of virus without clinical consequences was observed when the supply of breast milk anti-JHMV antibodies was terminated 5 days before infection. Immune reactions restricted by histocompatibility antigens do not play a crucial role in bestowing resistance. As neutralizing anti-JHM serum antibody titers of adult mice only rise sharply 5 to 7 days after infection, these results indicate that infection of adults can be arrested by immunological means but that, in addition, the rate of virus dissemination must be limited by other non-immunological mechanisms. PMID- 2987654 TI - A glycopolypeptide (gp 100) is the main antigen detected by HTLV-III antisera. AB - Sera from homosexuals and hemophiliacs in Germany were examined for antibodies to human T-lymphotropic retrovirus type III (HTLV-III) by an enzyme-linked immunosorbent assay (ELISA) against purified virus. ELISA positive sera were used to search by immunoprecipitation for HTLV-III related antigens in a persistently infected human T-cell line. A glycopolypeptide with a mol. wt. of 100 000 was regularly recognized by all positive sera. In analogy to glycosylation and high antigenicity of envelope polypeptides of other mammalian retroviruses, gp 100 seemed to be related to the env gen. Two polypeptides with mol. wts. of 24 000 and 22 000 probably representing viral core polypeptides were additionally detected by sera with high ELISA titers. PMID- 2987655 TI - [Analysis of trace elements present on dental surfaces]. PMID- 2987657 TI - Testing donors of organs, tissues, and semen for antibody to human T-lymphotropic virus type III/lymphadenopathy-associated virus. PMID- 2987656 TI - [Prosthetic rehabilitation in jaw resections followed by reconstruction of the palatal diaphragm]. PMID- 2987658 TI - D2-dopamine receptor-mediated inhibition of cyclic AMP formation in striatal neurons in primary culture. AB - Dopamine (DA) regulation of intracellular cyclic AMP formation in purified, intact striatal neurons in primary culture was examined. DA (EC50, 3 microM) and vasoactive intestinal polypeptide (VIP; EC50, 10 nM) stimulated cyclic AMP formation by 2- and 5-fold, respectively. In the presence of 0.1 microM forskolin (which was virtually ineffective alone), neurohormone efficacy was augmented; potency was unaffected. In the presence of 0.1 microM SCH 23390, a selective D1 antagonist, the DA dose-response curve was shifted rightward in a competitive manner. At low concentrations (0.01-1.0 microM), however, DA inhibited basal cyclic AMP formation. The inhibitory effect, but not the shift of the dose response curve, was blocked by 5 microM l-sulpiride, a selective D2 antagonist. At saturating concentrations of VIP (0.1-1.0 microM), no other neurohormone can further augment cyclic AMP formation. Under these conditions, increasing concentrations of DA resulted in a dose-dependent (IC50, 0.5 microM) inhibition of VIP-stimulated cyclic AMP synthesis. This effect was augmented in the presence of 0.1 microM SCH 23390 and blocked by 5 microM l-sulpiride. Sulpiride antagonism was stereospecific, with the l-isomer being 30-fold more potent than the d isomer. The rank order of potency for a series of dopaminergic agonists and antagonists at the receptor mediating attenuation of cyclic AMP formation suggests that it is of the D2 type. Furthermore, both DA and Met-enkephalin inhibition of cyclic AMP formation is lost after exposure of striatal neurons to islet activator protein. These findings suggest that a D2 receptor mediates the inhibition of intracellular cyclic AMP formation by DA in striatal neurons in primary culture, and may do so by an interaction with the inhibitory guanine nucleotide regulatory protein of adenylate cyclase. PMID- 2987659 TI - Kinetics of binding of membrane-active drugs to receptor sites. Diffusion-limited rates for a membrane bilayer approach of 1,4-dihydropyridine calcium channel antagonists to their active site. AB - Using the model of 1,4-dihydropyridine calcium channel-blocking drug binding to receptors in the cardiac sarcolemmal membrane, diffusion-limited rates of association were calculated for two distinct approaches. In the "aqueous approach," the drug reaches the receptor by diffusion through the bulk solvent, whereas in the "membrane approach," the drug partitions into the membrane bilayer and then diffuses laterally to a specific receptor site. Calculated rates for the membrane approach were approximately 3 orders of magnitude greater than those for the aqueous approach. The membrane approach diffusion-limited rate depends weakly on the sizes of the binding site, the drug molecule, and the vesicle, but depends strongly on ligand asymmetry. Although the measured binding rates for several 1,4 dihydropyridines were all slower than the calculated diffusion-limited rates for either model, other experimental data (such as very high partition coefficients and specific positions of these drugs in the membrane bilayer) suggest that the membrane approach is the most likely. These results have important implications for specifying critical characteristics of active 1,4-dihydropyridines. PMID- 2987660 TI - Effects of substituents on the cytosolic receptor-binding avidities and aryl hydrocarbon hydroxylase induction potencies of 7-substituted 2,3-dichlorodibenzo p-dioxins. A quantitative structure-activity relationship analysis. AB - The binding affinities of 16 7-substituted 2,3-dichlorodibenzo-p-dioxins for the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) cytosolic receptor protein from male Wistar rats have been determined. The EC50 value for each compound was estimated by competitive displacement of [3H]TCDD and the data illustrated that the differences between competitive ligands were dependent on the substituent (X) group. The EC50 value for 7-trifluoromethyl-2,3-dichlorodibenzo-p-dioxin was 1.95 X 10(-8) M and was greater than 1000-fold more active than 7-amino-2,3 dichlorodibenzo-p-dioxin (EC50 = 2.88 X 10(-5) M). Multiple parameter linear regression analysis of the data for 14 different compounds gave the following equation: log (1/EC50) = 1.24 pi + 6.11. This demonstrated that the binding affinity was linearly dependent on the lipophilicity (pi) of the 7-X-group. This contrasted with a comparable analysis of the substituent effects on the binding of 13 4'-substituted 2,3,4,5-tetrachlorobiphenyls to the cytosolic receptor which showed that the lipophilicity, electronegativity, and hydrogen-bonding capacity were important physicochemical determinants which facilitated binding to the receptor protein. These data suggest that the halogenated dibenzo-p-dioxins and biphenyls may interact with different binding sites on the receptor or they may bind to the same site but exert different conformational effects on the receptor protein. For the 7-X-2,3,-dichlorodibenzo-p-dioxins, there was not a rank order correlation between receptor-binding EC50 values and the induction of aryl hydrocarbon hydroxylase (AHH) or ethoxyresorufin O-deethylase in rat hepatoma H-4 II E cells in culture. However, the data could be correlated with an estimate of substituent width, the STERIMOL factor (B5), i.e., log (AHH) = 1.29 log (binding) + 2.19 delta B5 - 1.31 (delta B5)2 - 1.48. The importance of a steric factor in the correlation between receptor binding and AHH induction for substituted dibenzo-p-dioxins and halogenated biphenyls is consistent with a structure dependent conformational change(s) in the receptor protein:ligand complex after the initial binding event. Presumably, this latter process is associated with the steps involving interactions between the ligand:receptor complex and nuclear binding sites. PMID- 2987661 TI - Inhibition of utilization of hypoxanthine and guanine in cells treated with the carbocyclic analog of adenosine. Phosphates of carbocyclic nucleoside analogs as inhibitors of hypoxanthine (guanine) phosphoribosyltransferase. AB - In cell cultures treated with the carbocyclic analog of adenosine (C-Ado, (+/-) aristeromycin), the utilization of hypoxanthine and guanine has been observed to be blocked. In an attempt to define the mechanism of this inhibition, we have reexamined the metabolism of C-Ado and its effects on the metabolism of guanine and hypoxanthine. In cultures of L1210 cells, C-Ado at a concentration of 25 microM inhibited the utilization of hypoxanthine and guanine for nucleotide synthesis by more than 90% but produced little or no inhibition of the utilization of these bases in cultures of L1210/MeMPR cells which lack adenosine kinase and cannot phosphorylate C-Ado. In cultures of mammalian cells (L1210, HEp 2, and colon-26 cells), C-Ado was converted to the triphosphate (as previously observed) and also to the triphosphate of the carbocyclic analog of guanosine. The presence of coformycin in the medium at a concentration sufficient to inhibit AMP deaminase almost completely prevented the formation of carbocyclic GTP; thus, the deamination of C-Ado monophosphate is essential for the formation of phosphates of carbocyclic guanosine. Since hypoxanthine (guanine) phosphoribosyltransferase is known to be subject to end product inhibition, it was considered likely that phosphates of carbocyclic guanosine or carbocyclic inosine, present in C-Ado-treated cells, were responsible for inhibition of utilization of hypoxanthine and guanine. The 5'-phosphates of the carbocyclic analogs of inosine and guanosine were synthesized and found to be effective inhibitors of the phosphoribosyltransferase. Carbocyclic GMP was a better inhibitor than carbocyclic IMP and was also superior to GMP and IMP; the concentration of C-GMP that produced a 50% inhibition of GMP formation was approximately 1 microM. It is probable that the presence of phosphates of carbocyclic guanosine accounts for the inhibition of utilization of hypoxanthine and guanine in C-Ado-treated cells. PMID- 2987662 TI - [Oncoproteins--the products of oncogenes. Tyrosine protein kinases]. AB - The review summarizes the data mostly published in 1982 and 1983 concerning isolation, identification, cellular localization, deciphering of primary structure, and properties of the proteins coded by the retroviral oncogenes. A tentative classification of oncoproteins is suggested. The main part of the review is devoted to oncoproteins with tyrosine specific protein kinase activity. A hypothesis is proposed concerning the possible mode of action of these proteins on transforming cells. It is stressed in conclusion that the remarkable progress of our knowledge in this field has been mostly achieved due to recombinant DNA technique successfully and widely applied for studying oncogene structure and function. PMID- 2987663 TI - [Electron structure of the active center of paramagnetic hemoproteins from the data of low-temperature magnetic circular dichroism. High-spin ferric derivatives]. AB - The temperature dependences of magnetic circular dichroism (MCD) spectra of the high-spin fluoride complexes of metmyoglobin, methemoglobin and horse-radish ferriperoxidase have been studied and analysed in spectral region 250-1100 nm at temperatures from 2 to 300 K. Upon lowering of the temperature the increase of the MCD intensity was observed over all spectral regions studied, indicating the presence of the so called C-type effects. In contrast to other paramagnetic hemoproteins studied previously the significant changes of the shape of MCD spectra of fluoride complexes in the near UV B (pi--pi*)-band (Soret band) was observed at the lowest temperatures. The changes are explained by the superposition of effects of two types: the usual C-type effects which have the shape of absorption dispersion and of the derivative shape CA-type effects. The CA-type effects in the Soret band were analysed in terms of the theoretical model suggested previously. The data are in qualitative agreement with the prediction of the model. New information on the energies of the 4E and 4A2 excited states of the heme iron, on the orbital contribution into ground state paramagnetism and on the value of the pi--d pi-splitting, typical for B(pi--pi*)-state and ground Kramers doublet, were obtained on the basis of quantitative analysis of the MCD temperature dependences. The parameter D of zero-field splitting was estimated from MCD spectra and its value (6 cm-1) agrees well with that obtained by other methods. PMID- 2987664 TI - [Structural organization of the non-transcribed spacer of the rat ribosomal repeated sequence]. AB - Three types of repetitive sequences were found in the non-transcribed spacer of the rat ribosomal genes. Two internal repeats (100-150 bp long) flank the transcribed region encoding pre-rRNA. A number of internal repeats localized at approximately 300 bp upstream the transcription initiation site ranges from 1 to 9. This leads to heterogeneity in this region of the non-transcribed spacer. The non-transcribed spacer comprises of four regions with highly repetitive sequences. Two of these regions are localized 2.0-5.0 kb downstream the 3'-end of the 28S rRNA gene and the other two are localized 3.0-5.0 kb upstream the transcription initiation site. It is shown that these highly repetitive sequences are not only dispersed within the genome, but are found also in the family of extended repeats. One of the dispersed repeats localized upstream the transcribed region is reiterated more than 100 thousand times within the genome and is homologous to the mouse B2 sequence. The dispersed repeats downstream from the transcribed region are repeated 30-40 thousand times within the genome. The nucleotide sequences of the extended region of this sequence (approximately 1.0 kb) has typical blocks of (AC)n, (ACC)n, (GAG)n, (CTGT)n, (TAAC)n, (CCTG)n and (G)n types. Two palindromes and sufficiently long straight repeats were found. The structural-functional organization of the rat ribosomal repeating unit is a convenient model for studying the principles of organization of the genetic material in mammalian cells. PMID- 2987665 TI - [5S genes of the loach: determination of the primary structure of the transcription termination region and nontranscribed spacer]. AB - The primary structure of a 5S gene spacer has been determined by sequencing three cloned 5S rDNA fragments of the loach genome. The region of the spacer adjacent to the 3'-termini of the gene structural part was shown to comprise an AT-rich sequence (24 bp long) including an oligo (T)6-9 block corresponding to the terminator of RNA polymerase III. The results supported our previous data about the 5S rRNA precursor synthesis during the transcription of cloned 5S rDNAs injected into oocyte nuclei. The comparative sequence analysis revealed a homology between the spacer region from -54 to -26 bp and the 5'-termini (1-18 bp) and the 3'-termini (108-117 bp) stretches of the gene coding region. In addition, both the coding part and nontranscribed spacer of 5S DNA include highly diversed repeats which are homologous to the 3'-termini sequence (111-+8 bp) of the gene. Apparently the nontranscribed spacer of loach 5S genes derived in evolution from numerous replicated 5S genes as a result of subsequent base elimination and substitution in the most of the 5S rRNA coding sequences. PMID- 2987666 TI - [DNA-topoisomerases of animal cells]. AB - The properties and functions of the DNA topoisomerases are reviewed on the basis of the literature and the author's own data. The techniques of isolation and characterization of the covalent complexes of the topoisomerases with DNA are described. PMID- 2987667 TI - [Study of 2'-phosphodiesterase activity in cultured NIH 3T3 cells during activation of cAMP-dependent phosphorylation]. AB - A rapid and transient decrease in 2'-phosphodiesterase activity in NIH 3T3 mouse cells was observed after adrenaline addition. The decrease of activity was accompanied by an elevation of intracellular cAMP level. The 2'-phosphodiesterase activity changed similarly when cells sink deeper into the resting state. In the latter case, the fall of the enzyme activity was correlated with elevation of the activity of cAMP-dependent proteinkinase and, moreover, a considerable increase of the intracellular level of 2',5'-oligoadenylate was observed. Phosphorylation of proteins by cAMP-dependent proteinkinase in the cell lysate also produced a pronounced drop of 2'-phosphodiesterase activity. Exogenous 2',5'-oligo (A) treatment of the cells resulted in the rise of 2'-phosphodiesterase activity; actinomycin D prevented this effect. The data presented suggest the involvement of two different mechanisms in regulation of 2'-phosphodiesterase activity: cAMP dependent phosphorylation and induction of 2'-phosphodiesterase by 2',5' oligoadenylate. PMID- 2987668 TI - [Localization of regions of DNA attachment to the nuclear skeleton within chicken alpha-globin genes in functionally active and functionally inactive nuclei]. AB - Short DNA fragments remaining associated with the nuclear skeleton after nuclease treatment and high salt extraction of chicken erythroblasts and erythrocytes nuclei were used as hybridization probes in order to find the positions of DNA attachments to a nuclear skeleton inside the domain of alpha-globin genes. It is found that in erythroblasts nuclei a rather long region including coding sequences is associated with the nuclear skeleton. This region coincides with the region preferentially digested by DNAase I in chromatin. In erythrocytes nuclei, a specific attachment point is found in the region between 4500 and 300 bp upstream to pi-gene. Several attachments in this region are detected also in erythroblast nuclei, but they are not prominent in this case. The results are discussed in terms of the existence of stable (structural) and temporal (functional) attachments of DNA to the nuclear skeleton. PMID- 2987669 TI - [Comparative analysis of recombinant bacteriophages containing the sequences of the chromosomal ceruloplasmin gene in the rat]. AB - Three clones carrying the sequences of ceruloplasmin gene were isolated from the library of EcoRI fragments of rat chromosomal DNA cloned in Charon 4A vector. These clones were identified using, i) plaque hybridization with [32P] cDNA transcribed from highly purified rat ceruloplasmin (Cp) mRNA; ii) blot hybridization of the restriction fragments of recombinant DNAs with Cp cDNA and iii) hybridization selection of Cp mRNA followed by its cell-free translation. Oligo (dT)-primed cDNA transcripts of Cp mRNA having different length as well as cloned Cp cDNA isolated from rat liver cDNA library were used as hybridization probes for the study of mRNA-coding segments of Cp gene. The length of inserts in recombinant DNAs varied from 7.5 up to 12.3 megadaltons. EcoRI-fragments of Cp gene were mapped within recombinant DNA and their homology to each other was studied. PMID- 2987671 TI - Transient gene expression control: effects of transfected DNA stability and trans activation by viral early proteins. AB - The effects of trans-acting factors and transfected DNA stability on promoter activity were examined with chloramphenicol acetyl transferase (CAT) transient expression analysis. With cotransfection into CV-1P and HeLa cells, simian virus 40 T antigen, adenovirus E1a, and herpes-virus IE proteins were compared for their ability to trans-activate a variety of eucaryotic promoters constructed into CAT plasmids. T antigen and the IE protein were promiscuous activators of all the promoters tested [the simian virus 40 late promoter, the adenovirus E3 promoter, the alpha 2(I) collagen promoter, and the promoter of the Rous sarcoma virus long terminal repeat]. Conversely the E1a protein was specific, activating only the adenovirus E3 promoter and suppressing the basal activity of the other promoters. This specificity of activation by E1a contrasted with the high activity generated by all of the promoter-CAT plasmids when transfected into 293 cells, which endogenously produce E1a protein. Examination of transfected 293 cells determined that they stabilized much greater amounts of plasmid DNA than any other cells tested (CV-1P, COS, NIH-3T3, KB). Thus the high activity of nonadenovirus promoter-CAT plasmids in 293 cells results from the cumulative effect of basal promoter activity from a very large number of gene copies, not from E1a activation. This conclusion was supported by similar transfection analysis of KB cell lines which endogenously produce E1a protein. These cells stabilize plasmid DNA at a level comparable to that of CV-1P cells and, in agreement with the CV-1P cotransfection results, did not activate a nonadenovirus promoter-CAT plasmid. These results indicate that the stability of plasmid DNA must be considered when transient gene expression is being compared between cell lines. The use of relative plasmid copy numbers for the standardization of transient expression results is discussed. PMID- 2987670 TI - Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation. AB - We have analyzed the effects of transformation by Rous sarcoma virus on expression of types I and II collagen and fibronectin genes in vertebral chondrocytes and compared them with expression of these genes in skin fibroblasts. Transformed chondrocytes display a dramatically decreased amount of type II collagen RNA, which can account fully for the decreased synthetic rate of this protein. Paradoxically, these cells also display greatly increased amounts of type I collagen RNAs, which are translated efficiently in vitro, but not in the intact cells. We show here that the type I collagen RNAs in transformed chondrocytes are nearly indistinguishable from those found in skin fibroblasts, and that they clearly differ from the type I collagen RNAs found in normal chondrocytes. Transformed chondrocytes also display an increased amount of fibronectin RNAs, which can account fully for the increased synthetic rate of this protein. Thus, the effects of transformation by Rous sarcoma virus on type I collagen and fibronectin RNAs in chondrocytes are the opposite of those observed in fibroblasts, which display decreased amounts of these three RNAs. These data indicate that the effects of transformation on the genes encoding type I collagen and fibronectin must be modulated by host cell-specific factors. They also imply that the types I and II collagen genes may be regulated by different mechanisms, the type I genes being controlled at both transcriptional and posttranscriptional levels, and the type II gene being controlled primarily at the transcriptional level. PMID- 2987672 TI - Differential ability of a T-antigen transport-defective mutant of simian virus 40 to transform primary and established rodent cells. AB - The transforming potential and oncogenicity of a simian virus 40 (SV40) mutant affecting T-antigen (T-ag), SV40(cT)-3, was examined in an effort to dissect T-ag functions in transformation. SV40(cT)-3 has a point mutation at nucleotide 4434 that abolishes the transport of T-ag to the nucleus but does not affect its association with the cell surface. Transfection-transformation assays were performed with primary cells and established cell lines of mouse and rat origin. The efficiency of transformation for established cell lines by SV40(cT)-3 was comparable to that of wild-type SV40, indicating that transformation of established cell lines can occur in the absence of detectable amounts of nuclear T-ag. Transformation of primary mouse embryo fibroblasts by SV40(cT)-3 was markedly influenced by culture conditions; the relative transforming frequency was dramatically reduced in assays involving focus formation in low serum concentrations or anchorage-independent growth. Immunofluorescence tests revealed that the transformed mouse embryo fibroblasts partially transport the mutant cT ag to the cell nucleus. Transformed cell lines induced by SV40(cT)-3 did not differ in growth properties from wild-type transformants. SV40(cT)-3 was completely defective for the transformation of primary baby rat kidney cells, a primary cell type unable to transport the mutant T-ag to the nucleus. The intracellular localization of cellular protein p53 was found to mimic T-ag distribution in all the transformants analyzed. The mutant virus was weakly oncogenic in vivo: the induction of tumors in newborn hamsters by SV40(cT)-3 was reduced in incidence and delayed in appearance in comparison to wild-type SV40. These observations suggest that cellular transformation is regulated by both nuclear and surface-associated forms of SV40 T-ag. PMID- 2987673 TI - Surface T-antigen expression in simian virus 40-transformed mouse cells: correlation with cell growth rate. AB - Cell growth control appears to be drastically altered as a consequence of transformation. Because the cell surface appears to have a role in modulating cell growth and simian virus 40 (SV40)-transformed cells express large T antigen (T-Ag) in the plasma membrane, we investigated whether surface T-Ag expression varies according to cell growth rate. Different growth states were obtained by various combinations of seeding density, serum concentration, and temperature, and cell cycle distributions were determined by flow microcytofluorometry. Actively dividing SV40-transformed mouse cell cultures were consistently found to express higher levels of surface T-Ag and T-Ag/p53 complex than cultures in which cells were mostly resting. In addition, the T-Ag/p53 complex disappeared from the surface of tsA7-transformed cells cultured under restrictive conditions known to induce complete growth arrest (39.5 degrees C), although the surface complex did not disappear from other tsA transformants able to keep cycling at 39.5 degrees C. These results suggest that surface SV40 T-Ag or surface T-Ag/p53 complex, or both, are involved in determining the growth characteristics of SV40-transformed cells. PMID- 2987674 TI - Preferential expression of the c-fps protein in chicken macrophages and granulocytic cells. AB - We have studied the expression of the protein kinase activity of NCP98, the c-fps gene product, in several hemopoietic tissues of chickens as a function of the developmental stage of these organs. We found that in bone marrow, spleen, and bursa, maximum NCP98 kinase activity on a per-cell basis correlates with the peak of granulopoiesis in these organs. Furthermore, in a bovine serum albumin density gradient fractionation of bone marrow cells, granulocytic cells appeared to account for most of the NCP98 kinase activity. No correlation was found between the distribution of erythrocytic, lymphocytic, or thrombocytic cells and the distribution of the expression of NCP98 kinase activity. However, NCP98 protein and kinase activity were 10-fold higher in macrophages than in bone marrow. In addition, depletion by complement-mediated lysis of erythrocytic cells in bone marrow did not significantly reduce the total recovery of NCP98 kinase activity. These results argue for the specific expression of the c-fps gene product in granulocytic cells and macrophages. PMID- 2987675 TI - Nuclei of adenovirus 2-infected cells contain an RNA species that corresponds to an intron excised intact from mRNA precursors. AB - We used Northern and S1 nuclease analyses to identify a nuclear RNA species of the structure predicted for an intron excised from the precursor of adenovirus 2 E2A early mRNA. The structure of this excised intron demonstrated that the splicing of E2A mRNA can involve the removal of the intron sequences as single intact molecules. The concentration of the excised intron is 30 copies per nuclei, comparable to the levels of E2A polyadenylated splicing precursors, but 30-fold less than nuclear E2A mRNA. Late in infection, when the production of E2A early mRNA is dramatically elevated (Goldenberg et al., J. Virol. 38:932-939, 1981), the excess intron was not detected, indicating that either its stability or the mechanism of splicing is altered. We also identified a spliced nonpolyadenylated E2A nuclear RNA species with a structure similar to the mRNA, indicating that splicing may normally occur in the absence of polyadenylation. PMID- 2987676 TI - Alterations in chromatin structure associated with glucocorticoid-induced expression of endogenous mouse mammary tumor virus genes. AB - Alterations in the chromatin structure of endogenous mouse mammary tumor virus genes accompany glucocorticoid induction of viral RNA synthesis in the C57BL/6 T lymphoma cell line T1M1. These alterations are defined by the appearance of sites of DNase I hypersensitivity within proviral DNA in isolated nuclei, as well as by changes in the moderate nuclease sensitivity of entire proviral transcription units. Induced hypersensitive sites, termed type I, appear with a time course comparable to that required for induction of the rate of viral RNA synthesis and are maintained only in the continuous presence of hormone. Two such sites map to analogous positions in the 5' and 3' long terminal repeats of proviral DNA within, or very near, sequences that have been shown to comprise positions of specific binding of the glucocorticoid receptor in vitro and that are required for hormone-inducible transcription in vivo. A third type I site maps to another position of in vitro receptor binding near the 3' long terminal repeat. Some sites of DNase I hypersensitivity, termed type II, appear not to be markedly hormone dependent; two such sites are present in corresponding positions in each long terminal repeat. Comparison of the moderate DNase I sensitivity of mouse mammary tumor virus proviral DNA suggests that the three different endogenous units in T1M1 cells can be maintained in distinct chromatin conformations that are determined by factors related to the site of provirus insertion. It seems possible that altered chromatin conformations may reflect, or actually encode, important mechanistic features of these hormone-responsive genes. PMID- 2987677 TI - Ethidium bromide-induced loss of mitochondrial DNA from primary chicken embryo fibroblasts. AB - Chicken embryo fibroblasts in uridine-containing medium are inherently resistant to the growth-inhibitory effect of ethidium bromide. The drug was found to inhibit the incorporation of [3H]thymidine into mitochondrial DNA circular molecules. Mitochondrial DNA was quantitated by DNA-DNA reassociation kinetics with a probe of chicken liver mitochondrial DNA. A mean number of 604 copies of mitochondrial DNA per cell was found. This number decreased progressively in cells exposed to ethidium bromide, and by day 13 ca. one copy of mitochondrial DNA was detected per cell. When the cells were then transferred to drug-free medium, the number of copies increased very slowly as a function of time. On the other hand, analyses of DNA extracted from cell populations exposed to ethidium bromide for 20 or more days, with or without subsequent transfer to drug-free medium, revealed very little or no mitochondrial DNA by reassociation kinetics or by Southern blot hybridization of AvaI- or HindIII-digested total cellular DNA. As a result of the elimination of mitochondrial DNA molecules, the establishment of cell populations with a respiration-deficient phenotype was confirmed by measuring cytochrome c oxidase activity as a function of the number of cell generations and the absorption spectrum of mitochondrial cytochromes. PMID- 2987678 TI - Biochemical and immunological characterization of mutant L-M cells with altered levels of dibutyryl cyclic AMP-inducible alkaline phosphatase. AB - Phosphotyrosine-Sepharose 4B was synthesized and used to purify L-cell alkaline phosphatase. Antibodies to this enzyme interacted with the alkaline phosphatase of strains A-1-2 and A-3-3, mutants that express the enzyme constitutively. This and thermal stability studies suggest that these mutants contain the same alkaline phosphatase isozyme as their parent strain. PMID- 2987679 TI - Gene transfer method for transient gene expression, stable transformation, and cotransformation of suspension cell cultures. AB - A new method was developed to study transient gene expression, stable transformation, and cotransformation in suspension cells, such as mouse myeloma and erythroleukemia cells. This method involves attachment of cells to a concanavalin A-coated tissue culture dish, treatment of cells with DEAE-dextran to adsorb plasmid DNA to the attached cells, and finally treatment with a 40% solution of polyethylene glycol to facilitate the uptake of DNA by the cells. Plasmids pSV2cat and pSV2neo were used as markers to optimize the conditions for transient gene expression and stable transformation, respectively, of mouse myeloma and erythroleukemia cells. This method was successfully used to obtain cotransformants of mouse myeloma cells. PMID- 2987680 TI - The simian virus 40 sequences between 0.169 and 0.423 map units are not essential to immortalize early-passage rat embryo cells. AB - F8dl is a simian virus 40 early-region deletion mutant that lacks the sequences between 0.169 and 0.423 map units. We show that cloned F8dl DNA immortalized early-passage Fisher rat embryo cells with an efficiency that was about 20% of that of cloned wild-type simian virus 40 DNA. In contrast, we detected no immortalized colonies when we transfected the cells with DNA of five other early region deletion mutants that do not make stable truncated forms of T antigen. Since all five of these mutants have intact early- and late-region control sequences, we conclude that these control sequences are not sufficient for immortalization. Three of the mutants that did not immortalize did make a normal small t antigen, suggesting that the expression of this protein alone is not sufficient for immortalization of early-passage Fisher rat embryo cells. PMID- 2987681 TI - Characterization of pp60src phosphorylation in vitro in Rous sarcoma virus transformed cell membranes. AB - Phosphorylation of the src gene product pp60v-src was studied in plasma membrane fractions prepared from Rous sarcoma virus-transformed vole cells. Upon addition of [gamma-32P]ATP to isolated membrane vesicles, phosphate was incorporated into a 60,000-dalton polypeptide identified as pp60v-src. In the presence of vanadate, pp60v-src phosphorylation was stimulated ca. 30-fold. At low concentrations of ATP (1 microM), this reaction occurred almost exclusively on the carboxy-terminal 26,000-dalton region of pp60v-src. However, at higher ATP concentrations (100 microM), additional sites of phosphorylation were evident in the amino-terminal 34,000-dalton region. Kinetic analyses, performed under conditions in which ATP hydrolysis was minimal, revealed that the phosphorylation reaction at the carboxy terminus exhibited a higher Vmax and a lower Km for ATP than those occurring at the amino terminus. In addition, the amino-terminal region of pp60v-src was more rapidly dephosphorylated than the carboxy-terminal region. These results indicate that interaction of pp60v-src with the plasma membrane may limit the extent of amino-terminal phosphorylation by lowering the rate of the reaction and the affinity for the substrate while increasing its susceptibility to phosphoprotein phosphatases. We suggest that the use of transformed-cell membrane preparations provides a model system for studying the possible regulatory roles of phosphorylation and dephosphorylation on pp60v-src function. PMID- 2987682 TI - Cloning, sequencing, and functional analysis of oriL, a herpes simplex virus type 1 origin of DNA synthesis. AB - The herpes simplex virus type 1 genome (160 kilobases) contains three origins of DNA synthesis: two copies of oriS located within the repeated sequences flanking the short unique arm (US), and one copy of oriL located within the long unique arm (UL). Precise localization and characterization of oriL have been severely hampered by the inability to clone sequences which contain it (coordinates 0.398 to 0.413) in an undeleted form in bacteria. We report herein the successful cloning of sequences between 0.398 to 0.413 in an undeleted form, using a yeast cloning vector. Sequence analysis of a 425-base pair fragment spanning the deletion-prone region has revealed a perfect 144-base pair palindrome with striking homology to oriS. In a functional assay, the undeleted clone was amplified when functions from herpes simplex virus type 1 were supplied in trans, whereas clones with deletions of 55 base pairs or more were not amplified. PMID- 2987683 TI - Ubiquitin is a heat shock protein in chicken embryo fibroblasts. AB - Clones containing heat-inducible mRNA sequences were selected from a cDNA library prepared from polyadenylated RNA isolated from heat-shocked chicken embryo fibroblasts. One recombinant DNA clone, designated clone 7, hybridized to a 1.2 kilobase RNA that was present in normal cells and increased fivefold during heat shock. Clone 7 also hybridized to an RNA species of 1.7 kilobases that was present exclusively in heat-shocked cells. In vitro translation of mRNA hybrid selected from clone 7 produced a protein product with a molecular weight of approximately 8,000. Increased synthesis of a protein of similar size was detected in chicken embryo fibroblasts after heat shock. DNA sequence analysis of clone 7 indicated its protein product has amino acid sequences identical to bovine ubiquitin. In addition, clone 7 contains tandem copies of the ubiquitin sequences contiguous to each other with no untranslated sequences between them. We discuss some possible roles for ubiquitin in the heat shock response. PMID- 2987684 TI - Stimulation of expression of a herpes simplex virus DNA-binding protein by two viral functions. AB - We examined the expression and localization of herpesvirus proteins in monkey cells transfected with recombinant plasmids containing herpes simplex virus (HSV) DNA sequences. Low levels of expression of the major HSV DNA-binding protein ICP8 were observed when ICP8-encoding plasmids were introduced into cells alone. ICP8 expression was greatly increased when a recombinant plasmid encoding the HSV alpha (immediate-early) ICP4 and ICP0 genes was transfected with the ICP8 gene. Deletion and subcloning analysis indicated that two separate functions capable of stimulating ICP8 expression were encoded on the alpha gene plasmid. One mapped in or near the ICP4 gene, and one mapped in or near the ICP0 gene. Their stimulatory effects were synergistic when introduced on two separate plasmids. Thus, two separate viral functions can activate herpesvirus early gene expression in transfected cells. PMID- 2987686 TI - Tests for induction of dominant-lethal mutations and heritable translocations with tetrahydrocannabinol in male mice. PMID- 2987685 TI - TU elements: a heterogeneous family of modularly structured eucaryotic transposons. AB - We describe here a family of foldback transposons found in the genome of the higher eucaryote, the sea urchin Strongylocentrotus purpuratus. Two major classes of TU elements have been identified by analysis of genomic DNA and TU element clones. One class consists of largely similar elements with long terminal inverted repeats (IVRs) containing outer and inner domains and sharing a common middle segment that can undergo deletions. Some of these elements contain insertions. The second class is highly heterogeneous, with many different middle segments nonhomologous to those of the first-class and variable-sized inverted repeats that contain only an outer domain. The middle and insertion segments of both classes carry sequences that also are found unassociated from the inverted repeats at many other genomic locations. We conclude that the TU elements are modular structures composed of inverted repeats plus other sequence domains that are themselves members of different families of dispersed repetitive sequences. Such modular elements may have a role in the dispersion and rearrangement of genomic DNA segments. PMID- 2987688 TI - Cytogenetic changes induced in human diploid fibroblasts by tsA58 SV40 at permissive and restrictive temperatures. AB - Human diploid fibroblasts have been transformed by ts A58 SV40. At the permissive temperature, apparent chromosome and chromatic rearrangements were observed in a high percentage of cells and the frequency of SCE increased. If the transformed phenotype returned to normal at the restrictive temperature these alterations also returned to normal levels. Chromosome banding demonstrated many apparent chromosomal rearrangements in which diffuse staining material was joining intact chromosomes end-to-end and forming pseudostructural abnormalities. Homogeneously staining regions associated with gene amplification or virus-induced alterations in the coiling and stickiness of telomeric regions are possible mechanisms. PMID- 2987689 TI - Chromosomal aberrations induced by restriction endonucleases. AB - Restriction endonucleases (REs) are able to induce chromosomal aberrations in Chinese hamster ovary (CHO) cells. The G1 phase of the cell cycle seems to be especially sensitive for the induction of chromosomal aberrations by REs. The different capacities of REs to induce chromosomal aberrations are probably correlated with the number of recognition sites in the genome. PMID- 2987687 TI - Complementation of a pKM101 derivative that decreases resistance to UV killing but increases susceptibility to mutagenesis. AB - The drug resistance plasmid pKM101 makes Escherichia coli resistant to the lethal effects of ultraviolet (UV) irradiation and more susceptible to mutagenesis by a variety of agents. The plasmid operon responsible for increasing mutagenesis has been termed mucAB (Mutagenesis, UV and chemical). We have isolated a derivative of pKM101 called pGW1975 which makes cells more sensitive to killing by UV but which retains the ability of pKM101 to increase susceptibility to methyl methanesulfonate (MMS) mutagenesis. pGW1975 increases UV mutagenesis less than pKM101 in a uvrA+ strain but more than pKM101 in a uvrA- strain. muc- point and insertion mutants of pKM101 and pGW1975 complement to restore the plasmid mediated: (i) ability to reactivate UV-irradiated phage, (ii) resistance to killing by UV, and (iii) level of susceptibility to UV mutagenesis. We have identified a 2.0 kb region of pKM101 which is responsible for the complementation and which maps counterclockwise of mucAB. PMID- 2987691 TI - Metastatic small-cell carcinoma of the lung in a patient with AIDS. PMID- 2987690 TI - HTLV-III viremia in homosexual men with generalized lymphadenopathy. PMID- 2987693 TI - Fulminant liver failure induced by adenovirus after bone marrow transplantation. PMID- 2987692 TI - Human papillomavirus and carcinoma of the esophagus. PMID- 2987694 TI - Parvovirus infection: chance and investigation. PMID- 2987695 TI - An erythema infectiosum-like illness caused by human parvovirus infection. AB - In the spring of 1980, an epidemic of an illness that resembled erythema infectiosum occurred in Manitoba, Canada. We initiated prospective epidemiologic, clinical, and microbiologic studies of this illness among elementary-school children and their families. Initial microbiologic studies failed to identify the cause of the exanthem. After a similar illness associated with serologic evidence of human parvovirus infection occurred in London, stored specimens of 12 patients with exanthem were investigated for parvovirus infection. Eleven patients had parvovirus-specific IgM antibody, as did two family contacts and a teacher with nonexanthematous illnesses, and two asymptomatic family members. None of 28 children with measles or rubella had serologic evidence of recent parvovirus infection. Human parvovirus was detected by DNA hybridization and immune electron microscopy in the serum of one patient who later had a rash and in one unaffected family contact. Parvovirus DNA was also detected in the pharyngeal specimen of the teacher who was ill but did not have a rash. We conclude that human parvovirus infection can be asymptomatic or cause a variety of clinical manifestations, including nonexanthematous illness and an illness resembling erythema infectiosum. PMID- 2987696 TI - Evolution of tumours and the impact of molecular oncology. AB - It is generally accepted that tumours arise through the accumulation of several changes affecting the control of cell growth. Recent advances in molecular biology have made it possible to define some of these changes in molecular terms and to trace the steps by which certain tumours evolve. PMID- 2987697 TI - Chromosome Y-specific DNA in related human XX males. AB - Human 'XX males' are sterile males whose chromosomes seem to be those of a normal female. About 1 in 20,000 males has a 46, XX karyotype, and most cases are sporadic, that is, they are without familial clustering. It has long been argued that maleness in XX males may result from the undetected presence of a small, testis-determining fragment of the Y chromosome, and there is strong evidence for this in sporadically occurring XX males. Indeed, the genomes of three of four sporadic XX males tested were found to contain certain Y-specific DNA sequences. A pedigree in which three XX males occur has been interpreted as being consistent with autosomal recessive inheritance of maleness, and it has been argued that the basis of XX maleness in this family is fundamentally different from that in the sporadic cases. However, we report here that these related XX males, like the sporadic cases, contain portions of the Y chromosome. The portion of the Y chromosome present in one of the three XX males differs from that present in the other two. PMID- 2987698 TI - Selective inhibition of the anchorage-independent growth of myc-transfected fibroblasts by retinoic acid. AB - Fischer rat 3T3 (FR3T3) fibroblasts transfected with a cellular myc gene can be induced to grow and form colonies in soft agar by treatment either with epidermal growth factor (EGF) alone or with the combination of platelet-derived growth factor (PDGF) and type-beta transforming growth factor (TGF-beta). We now show that induction of anchorage-independent growth by each of these sets of growth factors involves different cellular pathways which can be distinguished by their sensitivity to retinoic acid. Colony formation induced by the combined action of PDGF and TGF-beta is 100-fold more sensitive to inhibition by retinoic acid than is colony formation induced by treatment of the myc-transfected cells with EGF. Moreover, retinoic acid (10(-8) M) is inhibitory for colony growth whenever TGF beta is present, regardless of whether the effects of TGF-beta are stimulatory, as occurs in the presence of PDGF, or inhibitory, as found in the presence of EGF. PMID- 2987699 TI - Association of phosphatidylinositol kinase activity with polyoma middle-T competent for transformation. AB - Polyoma middle-T antigen is required for viral transformation of cultured cells and for tumorigenesis in animals. Like many other transforming gene products, middle-T is bound to the membrane and has an associated tyrosine kinase activity in vitro. This activity seems to result from the interaction of middle-T with pp60c-src, the cellular homologue of the transforming gene product of the Rous sarcoma virus, pp60v-src (refs 3-5). Both pp60v-src (ref. 6) and another retrovirus transforming gene product, pp68v-ros (ref. 7) were shown recently to have an associated phosphatidylinositol (PI) kinase activity in vitro and to increase PI turnover in vivo. These results suggest that viral transformation may be directly connected to a complex network of second messengers generated from PI turnover. Here, we assayed for PI kinase activity in immunoprecipitates made with middle-T- or pp60c-src-specific antisera of cells infected with polyoma virus. A PI kinase activity was detected in those immunoprecipitates which contained middle-T. Studies of mutants of middle-T defective in transformation indicate a close correlation between PI kinase activity and transformation. PMID- 2987700 TI - A gelsolin-like Ca2+-dependent actin-binding domain in villin. AB - Villin is an actin-binding protein of relative molecular mass (Mr) 95,000 found in the core bundle of microfilaments in brush border microvilli from intestine. In physiological calcium concentrations (less than 1 microM), villin crosslinks actin filaments into bundles. However, in free calcium concentrations (greater than 1 microM), villin severs actin filaments into short pieces. To understand how villin can sever and bundle actin filaments, we are studying the molecular basis of villin-actin binding interactions by identifying important actin-binding domains in villin. Here, we report the purification and preliminary characterization of a 44,000-Mr fragment of villin which contains a calcium dependent actin-severing activity. In addition, the partial amino-acid sequence from the amino terminus of this fragment reveals homology with a 16-residue region near the amino terminus of gelsolin, an actin-severing protein found in many cells and sera. The sequence homology suggests a common structural basis for the calcium-regulated actin-severing properties shared by villin and gelsolin. PMID- 2987701 TI - Selfish DNA and the origin of introns. PMID- 2987702 TI - Segregation of pathways leading from area V2 to areas V4 and V5 of macaque monkey visual cortex. AB - V5 and V4 are areas of macaque monkey prestriate visual cortex that are specialized for involvement in different aspects of visual perception, namely motion for V5 (refs 1-4) and colour vision, with other possible functions, for V4 (refs 2, 5-9). Thus, it is unlikely that they should be fed the same information for further processing, yet both receive a strong input from patches of the upper layers of V2 (refs 10, 11), the area immediately adjoining the primary visual cortex, V1. V2, however, seems to comprise functionally distinct subregions, which can be revealed by staining the tissue for the mitochondrial enzyme cytochrome oxidase. Here we report that V4 and V5 are connected with separate cytochrome oxidase-defined subregions of V2, suggesting that cortical pathways dealing with motion and colour perception are segregated in their passage through V2, and reinforcing evidence for functional specialization in the visual cortex. PMID- 2987703 TI - Complex-unoriented cells in a subregion of primate area 18. AB - In primates, both the primary and secondary visual cortical areas can be subdivided histologically by staining for the mitochondrial enzyme cytochrome oxidase. In the primary visual cortex (area 17, the first cortical receiving area for visual information) these histological differences correspond to functional subdivisions, cytochrome-dark regions being concerned with information about colour and cytochrome-light regions concerned with form. Here we report that the second visual area, area 18, which receives its main cortical input from area 17 (refs 7,8), similarly has functional subdivisions that correspond to the cytochrome oxidase staining pattern. In area 18 the segregation between form and colour is maintained, reinforcing our notion that form and colour information follow parallel pathways. The specific differences between cells in areas 17 and 18 suggest that a possible step in hierarchical information processing is spatial generalization, analogous to the difference between simple and complex cells. PMID- 2987704 TI - Detection and sequence of mutations in the factor VIII gene of haemophiliacs. AB - The most common inherited bleeding disorder in man, haemophilia A, is caused by defect in factor VIII, a component in the blood coagulation pathway. The X chromosome-linked disease almost certainly stems from a heterogeneous collection of genetic lesions. Because, without proper treatment, haemophilia can be a fatal disease, new mutations are necessary to account for its constant frequency in the population. In addition, haemophilia A displays a wide range of severity, and some 15% of haemophiliacs generate high levels of antibodies against factor VIII ('inhibitor patients'). The present work elucidates the molecular genetic basis of haemophilia in some individuals. Using the recently cloned factor VIII gene as a probe, we have identified two different nonsense point mutations in the factor VIII gene of haemophiliacs, as well as two different partial deletions of the gene. Our survey of 92 haemophiliacs indicates no firm correlation between antibody (inhibitor) production and gross gene defects. PMID- 2987706 TI - [Spontaneous activity of the hair cells of the snail statocyst and changes in it as a result of intracellular stimulation]. AB - The activity of hair cells in the stotocyst of Helix lucorum was studied by means of intracellular recording. Maximal magnitude of input resistance of hair cell membranes was observed in the range of the injected hyperpolarizing current between 0 and 0.1 nA. Voltage noise was a specific type of hair cell activity. It was independent of synaptic activity and had no pacemaker nature. The voltage noise depended only on the contact between hair cell cilia and statoconia. Hair cell depolarization enabled to reveal the pacemaker properties of the receptor membrane. Spike-triggering zone was found to be located in the axon of hair cell. IPSP recorded in the hair cells of the preparations with the static nerve cut confirmed the existence of inhibitory interactions within the statocyst. PMID- 2987705 TI - The effect of the converting enzyme inhibitor HOE 498 on the renin angiotensin system of normal volunteers. AB - The converting enzyme inhibitor HOE 498 was evaluated in 12 normotensive male volunteers aged 21 to 26. The efficacy of single 5, 10 or 20 mg oral doses in blocking the pressor response to exogenous angiotensin I was tested in 3 of the subjects. All 3 doses of HOE 498 reduced the pressor response to exogenous angiotensin I to below 50% of control within 1,5 h following administration of the drug. Plasma renin and converting enzyme activity, blood angiotensin I, as well as plasma angiotensin II and aldosterone were measured serially before and up to 72 h following oral administration of a single dose of 2.5, 5, 10 or 20 mg of HOE 498 to groups of 5 volunteers each. As expected, blood angiotensin I levels and plasma renin activity rose while plasma converting enzyme activity, plasma angiotensin II and aldosterone concentration fell after administration of the drug. While the dose of 2.5 mg did not reduce plasma converting enzyme activity below 20% of control, the higher doses all resulted in plasma converting enzyme inhibition exceeding 90%. No side-effects were observed. It is concluded that in normal volunteers HOE 498 is an effective potent and long-acting converting enzyme inhibitor. Based on these preliminary findings it is expected that 5 mg HOE 948 will turn out to be adequate for therapeutic use. PMID- 2987708 TI - [Local nonuniformities in the syncytium of the horizontal cells of the retina in the turtle]. AB - Electrical coupling between horizontal cells of the turtle retina was investigated by means of two microelectrodes (current and recording ones) penetrating neighbouring cells at a fixed distance from each other. The morphological coupling was revealed by means of fluorescent dye Lucifer Yellow. The electrical coupling was confirmed between elements of similar type (L1- axonal terminals, or L2--cell bodies, or R/G type cells) and no coupling was found between elements of different types, though L1 and L2 are directly connected through thin axons. In the L1 syncytium the electrical coupling at small (less than or equal to 50 microns) but fixed distances between microelectrodes could differ several times depending on the minimal displacement of microelectrodes. This local nonuniformity of coupling can be explained on the basis of structural nonuniformities in the L1 (axon terminal) network. It is unlikely however that the structural nonuniformities can influence the functional properties of horizontal cell network when the retina is stimulated adequately (by light). PMID- 2987707 TI - [Hair cell interactions in the statocyst of the snail]. AB - Hair cell interactions within a statocyst of the snail (Helix lucorum) were studied by means of intracellular and extracellular recording. The obtained data demonstrated: some hair cells separated by less than or equal to 90 degrees on the circumference of the statocyst were electrically coupled; the electrical synapses were polarized because coupling coefficients within pairs of cells were not symmetrical (in one direction the coefficient was 10 times greater than in the other); some connections between hair cells located at less than or equal to 90 degrees from each other had electrochemical nature; excitatory chemical component of such interaction had a direction opposite to the effective electrical conduction; there were also inhibitory interactions between hair cells; some of them were monosynaptic chemical inhibitory ones (receptors were located approximately at 180 degrees from each other); others were polysynaptic weak interactions (hair cells were separated by more than 90 degrees-100 degrees on the circumference of the statocyst); all types of the interactions between hair cells were observed in preparations with the static nerve cut. PMID- 2987709 TI - [Synaptic responses of propriospinal neurons on stimulation of the locomotor strip of the dorsolateral funiculus of the cat]. AB - Stepping points in the dorsolateral funiculus of thoracic and cervical divisions of the spinal cord were found in mesencephalic cats. Synaptic responses of single neurons at level Th12-Th13 to microstimulation of these points were recorded extracellularly. Latencies of these responses increase when the distance between the stepping point and the site of the recording is over 20 mm, and paired stimuli have now to be delivered to evoke responses. Neurons responding orthodromically to stimulation of the stepping strip send their axons into the ventrolateral funiculus near the grey matter. Occurrence of antidromic responses among neurons sending their axons in the caudal direction is 37% in the average till the distance between the recording and stimulation sites is up to 40 mm. Half of neurons with synaptic responses to stimulation of the stepping point can be also excited by a train of 3-6 stimuli applied to the midbrain locomotor point. Revealed neurons may contribute to propagation of the activity to the stepping generator of the hindlimb. PMID- 2987710 TI - [Correlation between the plasticity of monosynaptic inputs of command neurons of Helix lucorum and the plastic properties of the chemoreceptive membrane during rhythmic stimulation]. AB - Degree of the EPSP depression in different command neurons of the snail defensive reaction during the repeated rhythmic stimulation of presynaptic neuron LPa7 is found to be diverse. A decrease in responses of neurons to microiontophoretic applications of acetylcholine is also significantly different. It is suggested that differences in the EPSP depression are closely connected with the heterogeneity of the chemoreceptive membrane properties. PMID- 2987711 TI - The study of the cytotoxic effect of anticancer drugs on human lung tumors transplanted to mice by use of diffusion chambers. AB - The effect of chemotherapeutic drugs (adriamycin, rubomycin, nitrosomethylurea, cyclophosphamide, vincristine, methotrexate) on the proliferative activity of cells of 19 human lung cancer (adenocarcinoma--8, squamous cell carcinoma--10 and small cell cancer--1) transplanted to mice in diffusion chambers was studied. Rubomycin and adriamycin showed highest activity while methotrexate was least effective. The cyclophosphamide, vincristine and nitrosomethylurea had the moderate and similar activity. The tumors of different histological types hade close sensitivity to the tested drugs. The 3H-thymidine incorporation inhibitory effect of drugs differed for squamous cell carcinoma and adenocarcinoma probably reflecting certain differences in cellular mechanisms of drug action in lung cancers of different histological types. The data on the cytotoxic activity of anticancer drugs against lung tumors were closed to known efficacy of these drugs in treatment of patients with lung cancer. The high activity of rubomycin in human lung tumors allow to hope for its high activity in patients. The results suggested that human lung tumors in diffusion chambers may be used in preclinical tests of antitumor drug activity against lung tumors. PMID- 2987712 TI - The in situ immunological reactivity and its significance in the clinical behavior of the cervical human papillomavirus lesions. AB - The in situ immunocompetent cell (ICC) infiltrates in 286 cervical punch biopsies derived from the women prospectively followed-up since 1981 (16 +/- 14 months, M +/- SD) for an established human papillomavirus (HPV) infection with or without coexistent cervical intraepithelial neoplasia (CIN) were characterized using the histochemical ANAE (acid alpha-naphthyl acetate esterase) staining. Only minor fluctuations in the relative proportions of ANAE T+, ANAE M+, and ANAE- cells were found between the three types of HPV lesions (flat, inverted and papillomatous condylomas), as well as between the lesions with varying degree of HPV-CIN. The percentage of ANAE- cells (mediators of humoral immune response) increases in parallel with the increasing density of the ICC infiltrate. The intensity of the local infiltrate was inversely correlated with the progression, but not with regression or persistence of the HPV lesions. The percentage of ANAE T+ cells was highest in the 32 (18.7%) HPV lesions progressed during the follow up, almost identical in the 88 (51.5%) persistent lesions and lowest in those 51 (29.8%) lesions showing clinical regression. The results are discussed in terms of the proposed immune surveillance functions attributed to the local ICCs according to the MALT (mucosal-associated lymphatic tissue) concept. Conclusion is drawn that no major imbalance in the ANAE- definable main populations of ICCs exist in the cervical HPV lesions, which readily could explain their divergent clinical behavior. Other factors (like HPV type, synergistic action of cocarcinogens or a deranged balance between the ICC subsets) should be searched for to elucidate this issue. PMID- 2987713 TI - Chemotherapy of metastatic islet cell carcinoma. Report of 4 cases and review of the literature. PMID- 2987714 TI - Serum angiotensin-converting enzyme increases during hemodialysis. Evidence for injury of the pulmonary vascular endothelium? AB - In 19 patients hemodialysis was associated with significant decreases in arterial oxygen tension and leukocyte count. After 240 min of dialysis serum angiotensin converting enzyme (ACE) corrected for hemoconcentration increased 14.3% (p less than 0.001) indicating injury of the vascular endothelium during hemodialysis. It is suggested that the increase in serum ACE is due to a complement-mediated sequestration of leukocytes in the pulmonary vasculature leading to injury of the vascular endothelium with interstitial edema and pulmonary dysfunction. Furthermore, it is suggested that serum ACE analysis may prove a new method to assess biocompatibility of materials used in devices for extracorporeal circulation. PMID- 2987715 TI - Effect of hemodialysis on red blood cell Na+-K+-ATPase activity in terminal renal failure. PMID- 2987716 TI - Erythrocyte membrane fluidity decreased in uremic hemodialyzed patients. AB - Erythrocyte membrane fluidity was studied by means of electron spin resonance in 15 uremic, hemodialyzed patients and 14 normal subjects. Erythrocyte membrane fluidity determined using a 16-nitroxide stearic acid spin label probe was of a significantly lower level in the uremic patients, when compared with normal control subjects. Alterations in molar ratios of membrane free cholesterol to phospholipid are probably not a principal factor contributing to this change in fluidity. Significant decreases of phosphatidylcholine and molar ratios of phosphatidylcholine to sphingomyelin were noted in the erythrocyte membrane of uremic patients, and these alterations may relate to the fluidity change. PMID- 2987717 TI - Increased serum angiotensin-converting enzyme in end-stage renal disease. PMID- 2987718 TI - Effects of acute oral administration of lithium compounds on multiple schedule performance in rats. AB - Adult male Sprague-Dawley rats were given acute oral administrations of lithium carbonate or lithium chloride at concentrations of 100, 200, and 300 mg/kg. Barpressing performance on a multiple fixed ratio 20/fixed interval 5 minute schedule was measured daily. The 200 and 300 mg/kg administrations of lithium chloride produced significant decreases in barpressing performance and all levels of lithium carbonate produced significant changes in performance. A 24 hour delay in the maximal behavioral effect of the 300 mg/kg dose of lithium carbonate paralleled previous findings showing delays in peak brain lithium levels after oral administration of lithium carbonate. PMID- 2987719 TI - Radiologic evaluation of the liver in the alcoholic patient. AB - It has been well documented that long-term abuse of alcohol leads to dysfunction of multiple organ systems of the body. The liver, which is the primary organ responsible for alcohol metabolism, is also a major target for damage. Cirrhosis of the liver is the seventh leading cause of death in the United States. The radiologist plays an important role in the evaluation and possibly in the treatment of the conditions which result from alcohol abuse. The advantages and limitations of various radiologic diagnostic modalities in the evaluation of alcoholic liver disease are presented and discussed. PMID- 2987720 TI - Demonstration of neurochemical tolerance following chronic barbital treatment. AB - Control rats (C) and rats rendered barbital dependent (BD) by the consumption of barbital chronically in their food for 6 weeks were treated with saline or 1 of 3 acute dosages of barbital 16 minutes before killing. Cyclic guanosine monophosphate (cGMP) levels and circulating levels of barbital were measured. Significantly higher circulating levels of barbital were required to produce a comparable suppression of cerebellar cGMP in the BD as opposed to the C rats. These data provide evidence for the development of tolerance to barbital in the cerebellar cGMP system during chronic barbital treatment. When barbital was abruptly withdrawn from BD rats, a profound elevation of cerebellar cGMP was observed to precede and outlast behavioral manifestations of barbital abstinence. Further understanding of the withdrawal induced aberrations in the mechanisms regulating cerebellar cGMP may give insight into the pathways involved in the manifestation of the barbital abstinence syndrome. PMID- 2987721 TI - Epinephrine-induced cyclic AMP production in skin fibroblasts from patients with dementia of Alzheimer type and controls. AB - Skin biopsies were obtained from six patients with dementia of Alzheimer type (DAT) and from three hospitalized age matched controls. Fibroblasts from these biopsies were grown in culture and compared for their growth characteristics and sensitivity to epinephrine with four cultures from age matched healthy individuals. The growth characteristics were similar in all three groups. The basal levels of cyclic AMP and the epinephrine-induced increase of cyclic AMP levels were also similar in control and DAT cells. PMID- 2987723 TI - Down-regulation of beta-adrenoceptors in rat cortex by repeated administration of desipramine, electroconvulsive shock and clenbuterol requires 5-HT neurones but not 5-HT. AB - Repeated administration to rats of desipramine (5 mg kg-1, twice daily for 14 days), clenbuterol (5 mg kg-1, twice daily for 14 days) or electroconvulsive shocks (5 ECS over 10 days) decreased the number of beta-adrenoceptors in the cortex. Injection of 5,7-dihydroxytryptamine into the lateral ventricle produced a selective depletion of cerebral 5-HT, particularly in the cortex. In confirmation of earlier studies, this lesion prevented the decrease in the number of beta-adrenoceptors produced by desipramine. However, it was found that this lesion also prevented the down-regulation of beta-adrenoceptors produced by repeated electroconvulsive shock and administration of clenbuterol. Treatment of rats with p-chlorophenylalanine (PCPA) produced a similar depletion of cerebral content of 5-HT. However, this depletion did not prevent the decrease in density of beta-adrenoceptors induced by desipramine or electroconvulsive shock. It is concluded that the modulation of the decrease in the number of beta-adrenoceptors by diverse antidepressant treatments is not by the action of 5-HT itself, but by the action of some other factor present at 5-HT terminals, which is removed by a neurotoxic lesion. PMID- 2987722 TI - Distribution of selected phospholipid modifying enzymes in rat brain microsomal subfractions prepared by density gradient zonal rotor centrifugation. AB - A rat brain P3 fraction enriched in ER derived microsomes was centrifuged through a 20-40% linear sucrose gradient in a Beckman Ti-14 Zonal rotor and 11 fractions were obtained. The distribution of marker enzyme activities and protein were determined in these 11 subfractions. NADPH-Cytochrome C reductase, choline phosphotransferase were employed for endoplasmic reticulum, Na+,K+-ATPase, 5' nucleotidase, and acetylcholinesterase were employed for plasma membrane, 2',3' cyclic nucleotide phosphohydrolase was employed for myelin. The bulk of the protein was recovered in the 24-34% sucrose fractions, Na+,K+-ATPase, 5' nucleotidase, and acetylcholinesterase were in the 22-38% sucrose fractions while NADPH-cytochrome C reductase and CNPase were enriched in the 20-22% sucrose fractions. The ethanolamine and the serine base exchange activities had a bimodal distribution, with highest specific activities in sucrose fractions 32-34% and 20 24%. Choline base exchange activity was nearly undetectable in all the fractions. The specific activities of CDP-choline phosphotransferase, and phospholipid-N methyltransferase were highest in the 20-22% sucrose fraction. Phospholipid-N methyltransferase activity was significantly stimulated in the presence of exogenous phospholipid acceptors as phosphatidylethanolamine or phosphatidylmonomethylethanolamine or phosphatidyldimethylethanolamine, however, the greatest response was with phosphatidylmonomethylethanolamine. The rat brain P3 fraction yielded a population of a membrane at the light end of the sucrose gradient which has a buoyant density similar to myelin but seemed to be enriched with NADPH cytochrome C reductase and phospholipid modifying enzymes. This is in contrast to liver microsomes submitted to a similar fractionation. PMID- 2987724 TI - The effects of benzodiazepines on spinal homosynaptic depression. AB - Clonazepam (0.5 mg/kg, i.v.) changed the characteristic pattern of the exponential decline of the monosynaptic responses, the early tetanic rundown, evoked by trains of 10 stimuli (2, 5 or 10 Hz) applied to either the biceps semitendinosus or triceps surae nerve, and recorded from the ventral root in spinal cats. In the case of the biceps-semitendinosus, clonazepam did not affect the first monosynaptic response or the last five monosynaptic responses forming the plateau, while the second monosynaptic response was markedly depressed, especially at the higher frequencies tested. The triceps surae reacted differently to the administration of clonazepam, in that the first response was increased and the amount of depression of the second response was lessened, with no change of the plateau. All the effects of clonazepam were reversed by the benzodiazepine antagonist, ethyl 8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H imidazo[1,5-a] [1,4]benzodiazepine-3-carboxylate (Ro15-1788; 5 mg/kg, i.v.), which alone had no effect of its own on any parameters, suggesting that the effects of clonazepam were mediated by central benzodiazepine receptors. Diazepam (1.0 mg/kg, i.v.), caused the same changes in the homosynaptic depression of the biceps-semitendinosus pathway as did clonazepam, but increased the plateau instead of the second response in that of the triceps surae pathway. PMID- 2987725 TI - A GABAergic component in homosynaptic depression in the spinal monosynaptic pathway. A requirement for action of benzodiazepines. AB - Spinal monosynaptic responses, evoked by repetitive stimulation, undergo homosynaptic depression the pattern of which is altered by 0.5 mg/kg of clonazepam. The dependence of this effect of clonazepam on the GABAergic system was examined in spinal unanaesthetized cats. Topical application of bicuculline to the spinal cord did not change any feature of the homosynaptic depression in the biceps-semitendinosus (BST) or triceps surae (TS) monosynaptic pathway but antagonized the action of clonazepam. Semicarbazide (200 mg/kg, i.v.) also prevented the effect of the benzodiazepine but alone had actions of its own. Evidence is presented that clonazepam influenced homosynaptic depression of the biceps-semitendinosus pathway by lengthening the primary afferent depolarization (PAD). This prolongation of the primary afferent depolarization did not last for the entire duration of the train as primary afferent depolarization also underwent depression. Therefore later responses in the train were unaffected by clonazepam. Homosynaptic depression of the triceps surae pathway was not similarly affected because activation of triceps surae afferents does not cause significant depolarization of its own afferents. It is suggested that the enhancement of GABAergic transmission at least partially underlies the effect of clonazepam on homosynaptic depression. PMID- 2987726 TI - Grooming behavior induced by ACTH involves cerebral cholinergic neurons and muscarinic receptors. AB - The grooming behavior induced by intracerebroventricular administration of ACTH was specifically antagonized by the muscarinic receptor antagonists, atropine and scopolamine. The antagonism occurred in a dose-dependent manner with an ED50 of 0.2 mg/kg (i.p.) for atropine, and 0.03 mg/kg (i.p.) for scopolamine. Neither methylscopolamine nor methylatropine, injected intraperitoneally affected grooming induced by ACTH (except for a small inhibition at the largest dose of methylscopolamine), but pronounced inhibition occurred when these drugs were administered intracerebroventricularly. The grooming induced by ACTH was also inhibited by prior intracerebroventricular administration of hemicholinium-3, a cholinergic antagonist that acts presynaptically by inhibiting the uptake of choline and hence, the synthesis of acetylcholine. These data suggest a role for cholinergic neurons and muscarinic receptors in the CNS in grooming behavior induced by ACTH. PMID- 2987727 TI - Exposure to rotating magnetic fields alters morphine-induced behavioral responses in two strains of mice. AB - An exposure for 60 min to a 0.5 Hz rotating magnetic field (1.5-90 G) significantly reduced the day-time analgesic and locomotory effects of morphine (10 mg/kg) in CF-1 and C-57BL strains of mice, respectively. Exposure to lower intensity 60 Hz magnetic fields (0.-1.0 G) had no effect on analgesia induced by morphine. The reduction in responsiveness to morphine after exposure to the greater intensity rotating field was not evident 24 hr later. No changes were seen in the latencies of basal thermal responses or levels of activity of saline treated mice exposed to the magnetic stimuli. PMID- 2987728 TI - Cardiovascular effects of tetanus toxin after systemic and intraventricular administration in rats. AB - A single intraperitoneal injection of tetanus toxin (50,100 and 200 minimum lethal dose per rat) produced a significant and dose-dependent increase in systemic blood pressure and heart rate in conscious rats. On the contrary, tetanus toxin, given directly into the third cerebral ventricle (20 and 40 minimum lethal dose), produced a gradual and dose-dependent decrease in blood pressure accompanied by bradycardia. In conclusion, the present findings show that tetanus toxin is able to affect cardiovascular activity and it is suggested that this may be due to an interference with central GABAergic mechanisms. PMID- 2987729 TI - A pharmacological study of the effect of carbamazepine on neuromuscular transmission in the rat diaphragm. AB - The effects of carbamazepine (0.042-0.42 mM) on neuromuscular transmission were studied on the isolated rat phrenic nerve diaphragm preparation using standard pharmacological and electrophysiological methods. Carbamazepine decreased (1) the antidromic activity of the phrenic nerve, (2) the amplitude of the endplate potential (EPP) and miniature endplate potential (MEPP), (3) the quantal content of the endplate potential, (4) the indirectly-elicited twitch tension, (5) the muscle contracture in chronically denervated muscle induced by acetylcholine (ACh) and (6) the amplitude of the compound phrenic nerve action potential, in a concentration-dependent manner. The antidromic activity of the phrenic nerve was the most affected, while the phrenic nerve compound action potential was least affected. However, the IC50 for carbamazepine (the concentration of carbamazepine that inhibited 50% of the response) was in the same order of concentration, i.e. 0.11-0.3 mM. Compared with the effect of carbamazepine on the indirectly-elicited twitch tension with its actions described above, it is concluded that carbamazepine interfered with the neuromuscular activity by inhibiting pre- and postsynaptic process and conduction in the phrenic nerve. PMID- 2987730 TI - Effects of lithium in vitro on noradrenaline-induced cyclic AMP accumulation in rat cortical slices after reserpine-induced supersensitivity. AB - Lithium in vitro at concentrations of 1-2 mM and above inhibited cyclic AMP accumulation due to noradrenaline in cerebral cortical slices prepared from both control rats and rats rendered supersensitive for the noradrenaline response by intraperitoneal injection of reserpine. There was no difference in the concentration-dependence of lithium inhibition between control and reserpine treated rats. PMID- 2987732 TI - In vivo evaluation of mu, delta and kappa opioid receptor agonists on spinal 5-HT metabolism in the rat. AB - Morphine was found to elevate spinal dorsal horn 5-hydroxyindole acetic acid (5 HIAA) in a dose-dependent manner. These actions also were naloxone-reversible. In marked contrast other mu, delta and kappa opioid receptor agonists did not alter 5-HIAA levels. These studies clearly indicate that descending spinal 5 hydroxytryptamine (5-HT) pathways are not essential for opiate analgesia. PMID- 2987731 TI - Studies in vivo with ICI 174864 and [D-Pen2, D-Pen5]enkephalin. AB - We studied the in vivo pharmacology of a selective agonist (DPDPE) and a selective antagonist (ICI 174864) at delta opioid receptors. ICI 174864 (10 micrograms icv) caused postural abnormalities, barrel rotation and hypothermia in rats. DPDPE induced behavioural arousal (at 75 micrograms icv) and barrel rotation (at 125 micrograms) in rats. ICI 174864 (10 micrograms icv) attenuated acetic acid induced writhing in mice. This action was antagonized by naloxone (10 but not 2 mg/kg s.c.). A lower, non-agonist dose of ICI 174864 (5 micrograms) antagonized DPDPE (3 micrograms icv) in this test without affecting DAGO (0.0006 micrograms icv), a selective agonist at mu receptors. In the mouse tail flick test, ICI 174864 (10-50 micrograms icv) did not significantly antagonize the agonist actions of DPDPE (40 micrograms icv) or DAGO (0.3 micrograms icv). At 10 50 micrograms icv, ICI 174864 had no marked effect on gastrointestinal transit in mice. ICI 174864 (25 micrograms icv or 20 mg/kg s.c.) did not interact with mu opioid receptors in mice rendered physically dependent on morphine. PMID- 2987733 TI - N-tetrahydrofurfuryl benzomorphans: comparison of a kappa and an agonist/antagonist analgesic. AB - The N-tetrahydrofurfuryl benzomorphan, MK-2034 is a prototype kappa agonist analgesic. Dehydrogenation and addition of a methyl group into the N-furfuryl side chain (MR-1353) results in a 2-fold decrease in analgesic potency, a 12-fold decrease in affinity for the mu receptor, a 5-fold decrease in affinity for the delta receptor and a 60-fold decrease in affinity for the kappa receptor. Both compounds possess intermediate sodium ratios for 3H-naloxone binding; however, unlike MR-2034, MR-1353 possesses a bell-shaped dose response curve for elevations in striatal dopamine metabolites. These data indicate that MR-1353 is an agonist/antagonist analgesic and suggest that the high kappa affinity of MR 2034 is responsible for the potent analgesia. PMID- 2987734 TI - Sigma opioid receptors; SKF-10,047 update. AB - We found that (-)-SKF-10,047 blocks EEG and behavioral effects of morphine in the naive rat, precipitates withdrawal in morphine-dependent rats, produces physical dependence as evidenced by naloxone-induced withdrawal, and displaces [3H] dihydromorphine from brain homogenates. (+)-SKF-10,047 did not produce dependence upon chronic treatment, and it did not displace [3H] dihydromorphine from brain homogenates. Such pharmacodynamic dissociation of SKF-10,047 effects suggests an association of sigma receptors with psychotogenic, but not opioid characteristics. The latter are most likely mediated by mu or kappa receptors. PMID- 2987735 TI - Evidence for opiate receptor involvement in the consumption of a high palatability diet in nondeprived rats. AB - Nondeprived adult rats were familiarized with a highly palatable diet (powdered small animal diet mixed with sweetened condensed milk and water). The palatability of food was such that it induced vigorous feeding responses, 15-20 g food consumed within the first 30 min of access. In partially-satiated male rats, the kappa receptor agonists EKC and U-50,488 (subcutaneously administered) produced large increases in food consumption in the first 30 min of access, post injection. In experiments with naloxone and WIN 44,441-3, we found that the effects of naloxone (0.01-10 mg/kg; S.C.) were crucially dependent on the sex and dietary history of the animals. Male, obese rats were most sensitive to naloxone's anorectic effect. Lean females were completely insensitive. WIN 44,441 3 (0.01-10 mg/kg, S.C.) had no effect on food intake in any group of animals. PMID- 2987736 TI - Selective attenuation of sweetened milk consumption by opiate receptor antagonists in male and female rats of the Roman strains. AB - Male and female rats of the three Roman strains (Roman High-, Roman Low-, and Roman Control Avoidance; RHA, RLA and RCA, respectively) were familiarized with a highly palatable sweetened milk in a daily 30 min test. The animals were never food- or water-deprived prior to the test. Daily milk intake stabilised at a high level before drug tests were initiated. Effects of naloxone, diprenorphine, WIN 44,441-3, MR2266, MR2267, and ICI 154129 on milk consumption were investigated. Naloxone, diprenorphine and MR2266 each had comparable anorectic effects across strains and sexes. WIN 44,441-3 was relatively ineffective; MR2267 and ICI 154129 were without effect on milk consumption. PMID- 2987737 TI - Enkephalins and endorphins: activation molecules for the immune system and natural killer activity? AB - This article summarizes some of the immunological activities of enkephalins and endorphins. Human and animal lymphocytes possess receptors for endorphins and enkephalins. These opioid peptides enhance T cell mitogen response. They decrease in vitro antibody formation. Endorphins and enkephalins will enhance human natural killer activity. This effect may be mediated by several mechanisms including the production of factor(s) enhancing natural killer activity by lymphocytes incubated in presence of opioid peptides. All these results suggest that opioid peptides may be activation molecules for the immune system and as such immunomodulatory agents. PMID- 2987738 TI - Relative affinities of the quaternary narcotic antagonist, N-methyl levallorphan (SR 58002), for different types of opioid receptors. AB - The relative affinities for different subtypes of opioid receptors (mu, kappa and delta) of the peripheral narcotic antagonist N-methyl levallorphan (SR 58002) have been studied in two in vitro smooth muscle systems (guinea-pig ileum and rabbit vas deferens) and by binding studies (guinea-pig brain and cerebellum membranes) using selective tritiated ligands. All the evidence obtained indicates that SR 58002 is a pure antagonist with relative affinity for mu receptors vs kappa and delta superior to that of the parent tertiary compound, levallorphan. PMID- 2987740 TI - Effects of opiates on urine output in the water-loaded rat and reversal by beta funaltrexamine. AB - The effects of several mu and kappa opioid receptor agonists on urinary excretion were examined in the water-loaded rat. Mu agonists induced anti-diuresis whereas kappa agonists caused diuresis. Furthermore, high efficacy kappa agonists eg. U50, 488H and tifluadom, produced marked diuretic effects, whereas kappa partial agonists eg. nalorphine, produced low maximum diuresis. Compounds having activity at both mu and kappa receptors eg. ethylketocyclazocine(EKC) and MR2034, produced a biphasic effect, an initial anti-diuretic effect followed by a more sustained diuretic effect. The irreversible mu receptor antagonist, beta-funaltrexamine (beta-FNA) antagonized the anti-diuretic effects elicited by fentanyl, pentazocine and buprenorphine and the initial anti-diuretic effect induced by EKC but did not inhibit the diuretic effects induced by U50, 488H and EKC. Thus, the use of beta-FNA in the water-loaded rat provides us with a valuable in vivo test which differentiates both mixed and selective opioid receptor agonists. PMID- 2987739 TI - Studies in vitro with ICI 174,864, [D-Pen2, D-Pen5]-enkephalin (DPDPE) and [D Ala2, NMePhe4, Gly-ol]-enkephalin (DAGO). AB - The interactions of a proposed, selective delta receptor antagonist (ICI 174,864) and selective agonists at mu and delta receptors, [D-Ala2, NMePhe4, Gly-ol] enkephalin (DAGO) and [D-Pen2, D-Pen5]-enkephalin (DPDPE), respectively, have been studied using the electrically-stimulated mouse isolated vas deferens (MVD) and the guinea-pig isolated ileum (GPI). Incubation of increasing concentrations of ICI 174,864 (10,30,100 and 300 nM) produced a dose-related and parallel rightward displacement of the DPDPE dose-response curve in the MVD. In contrast, ICI 174,864 (300-3000 nM) failed to affect the DAGO dose-response curve in the same tissue. Analysis of the DPDPE-ICI 174,864 interaction in the MVD using the pA2 method revealed a Schild plot slope of -0.68 suggesting the involvement of more than one population of receptors. ICI 174,864 (300 nM) failed to antagonize DPDPE in the GPI at doses up to 30 microM. These results suggest that (a) ICI 174,864 acts as a selective delta antagonist in the MVD; (b) DPDPE interacts with mu receptors in the MVD but only at very high concentrations, and (c) delta receptors appear not to be of functional importance in the GPI. PMID- 2987742 TI - Changes in releasability of ACTH and beta-endorphin with chronic stress. AB - The activation of the hypothalamic pituitary adrenal axis by stress is well known. Using inescapable intermittent footshock as a stressor in rats, we have previously demonstrated a rise in circulating plasma Beta-endorphin/Beta-LPH which parallels the rise in plasma ACTH, the primary POMC derived peptides released by anterior lobe. In addition, the rise in ACTH is accompanied by approximately a tenfold rise in plasma corticosteroids. Short term anterior lobe pituitary cultures from rats who have received inescapable intermittent footshock for 30 minutes show a blunted dose response curves to the ACTH releasing secretagogues arginine vasopressin (AVP) and ovine corticotropin releasing factor (oCRF). Similarly a blunted dose response curves to secretagogues can be seen by either the addition of dexamethasone (0.5 nM) to the culture medium or pretreatment of the rats with 1 mg dexamethasone intraperitoneally 90 minutes prior to decapitation. Thus, glucocorticoids may play a role in the blunted response to secretagogues seen in anterior lobe cultures from acutely stressed rats. We now report that chronically stressed rats exhibit increased releasability of ACTH and Beta-endorphin/Beta-LPH products by oCRF, suggesting an increase of the peptides in the releasable pool. PMID- 2987741 TI - Postnatal development of opioid systems in rat brain. AB - Immunocytochemical and receptor autoradiographic techniques have been utilized to examine the postnatal development of opioid peptides and receptors. The pattern of met-enkephalin-like immunoreactivity was very similar at birth to the adult, increasing only in intensity throughout postnatal development. In contrast, beta endorphin-like immunoreactivity (BLI) in neonate was strikingly different from that of adult. In particular, cells and fibers were observed in germinal zones, structures present only in the neonate. The distribution of dynorphin-B-like immunoreactivity at birth was intermediate in its similarity to that of adult. Certain terminal fields were present, others developed postnatally, and others disappeared with age. In parallel studies, autoradiographic maps of mu, delta and kappa opioid receptor subtypes were generated to determine whether any correspondences existed between the developmental distribution of opioids and their receptors. Although significant overlap existed, no simple one-to-one relationship was observed. The differential localization of peptides and receptors in neonatal and adult brain suggests that opioids may subserve distinct functions in the neonate. In particular, the presence of BLI in the germinal zones, where postnatal neurogenesis occurs, implicates opioid systems in regulation of neuronal cell division. PMID- 2987743 TI - Biochemical changes due to narcotic receptor mediated phenomenon. PMID- 2987744 TI - Dynorphin receptor in the blood vessel. AB - Using (3H)etorphine, (3H)E, in binding studies, the KD and Bmax for rabbit mesentery and aorta membrane preparations were 0.61 nM and 0.17 fmol/mg tissue respectively, while it was 0.30 nM and 12 fmol/mg tissue in the brain. The IC50 of dynorphin (1-13) (D1-13) for displacing (3H)E binding in the blood vessel was 20 +/- 2.8 nM (S,E,M,), while PLO17, D-ala2-D-leu5-enkephalin (DADLE) and met5 enkephalin-arg6-phe7 showed very weak inhibition (IC50 greater than 1000nM) though they displaced (3H)E binding very well in the brain. In vitro study showed that D1-13 inhibited electric field stimulation induced vasoconstriction with an IC50 of 53 +/- 12 (rabbit ear artery) and 510 +/- 120 (dog mesenteric artery)nM. Such effect was partially reversed by 1 microM of naloxone. D-ala2-met5 enkephalin and metorphamide displayed much weaker inhibition and DADLE was completely ineffective at doses up to 1 microM. D1-13 did not antagonize noradrenaline (NA) induced vasoconstriction, while phentolamine could abolish vasoconstriction induced either by stimulation or by NA. The result suggests that D1-13 acts presynaptically on neuronal kappa receptor in the blood vessel and inhibits NA release, thus causes vasodilation. PMID- 2987745 TI - The effects of neurotensin and [D-Tyr11]-NT on the hyperactivity induced by intra accumbens administration of a potent dopamine receptor agonist. AB - The effects of neurotensin on the strong and persistent hyperactivity induced in rats by intra-accumbens administration of ADTN, a potent dopamine agonist, were examined. Neurotensin was administered intraventricularly as well as bilaterally into the accumbens. With both routes of administration neurotensin significantly decreased the hyperactivity produced by ADTN. However, important differences in doses required to produce this effect were noted between the two routes of administration. Whereas intraventricular injection of doses as small as 0.05 micrograms neurotensin was sufficient to reduce hyperactivity, bilateral intra accumbens administration of at least 1.8 micrograms was required to replicate the effect. ADTN induced hyperactivity was also significantly decreased by intraventricular and intra-accumbens injections of the structural analog [D Tyr11]-NT. In both routes of administration, the inhibitory action of the analog was more persistent than that observed with neurotensin. As was the case for neurotensin, intraventricular administration of [D-Tyr11]-NT was more potent than intra-accumbens injections. Finally, the results of a preliminary experiment indicate that neurotensin injected intraventricularly can also decrease hyperactivity elicited by intra-accumbens administration of dibutyryl cyclic AMP. Taken together, the results of the present study demonstrate that neurotensin can affect hyperactivity elicited by a strong and persistent activation of mesolimbic dopamine receptors or by stimulation of events beyond these receptors. The observed greater efficacy of intraventricularly administered neurotensin in decreasing ADTN induced hyperactivity suggests an action of the peptide on regions distant from the accumbens, probably on efferent outputs of mesolimbic stimulation. PMID- 2987746 TI - Increased affinity of dimeric enkephalins is not dependent on receptor density. AB - The equilibrium binding and dissociation kinetics of the enkephalin dimer bis-(D Ala2-D-Leu5-enkephalin)-ethylenediamide (designated DPE2) to neuroblastoma glioma NG108-15 cells were investigated and compared with the monomers D-Ala2-D-Leu5 enkephalin (DADL) and D-Ala2-Leu5-enkephalinamide (DALEA). Binding was studied after exposure of the membrane to increasing concentrations of the irreversible delta receptor selective ligand FIT in order to decrease the density of binding sites on the cell membrane. The increased affinity of DPE2 did not revert to that of the monomer DADL by this reduction of binding sites. Similarly, the dissociation of DPE2 did not approach that of the monomer DALEA in the presence of 1 microM DALEA. These data strongly suggest that crosslinking does not occur, and fail to confirm the hypothesis that dimers with short spanning chain length aid the clustering of receptors. We postulate: 1) If the dimer binds to a bivalent binding site, the monovalent binding state of our bivalent ligand may not exist to an appreciable extent, and 2) the bivalent ligand cannot bind when the binding site is irreversibly blocked by a monovalent ligand. PMID- 2987747 TI - Brain biopsy in cases of neonatal herpes simplex encephalitis. AB - Neonatal herpes simplex encephalitis (HSE) can represent a difficult diagnostic problem when it occurs without concomitant mucocutaneus lesions and usually requires brain biopsy for diagnosis. Asymptomatic for the initial 2 to 4 weeks of life, the three infants we describe with localized HSE came to medical attention only because they developed persistent seizures and other nonspecific symptoms. Lumbar spinal fluid obtained from these children at clinical presentation showed an encephalitic pattern. Radionuclide brain scans revealed focal uptake of isotope in a variety of cortical areas, and electroencephalograms (EEGs) demonstrated repetitive, high amplitude, polyphasic sharp waves arising from analogous regions. Computed tomography (CT) showed nonspecific ill-defined areas of low density or contrast enhancement that did not correlate well with radionuclide, EEG, or clinical findings in two neonates. No infant had predominant temporal lobe involvement. Because these data suggested a multifocal, encephalitic process, all three infants underwent brain biopsy. A widespread infiltration of leukocytes and macrophages was observed in each specimen, and abundant intranuclear inclusions were present. Electron microscopy revealed abundant herpesvirus particles, and herpes simplex virus (HSV) was subsequently isolated from each sample. From our observations and our review of the literature, we propose the following criteria as indications for brain biopsy: Brain biopsy is warranted to rule out HSE when a neonate presents with seizures, cerebrospinal fluid mononuclear pleocytosis with a negative gram stain, and focal, cortical disease on EEG and radionuclide scan.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2987748 TI - Cerebral glioblastomas can be cured! AB - In 1959, a 30-year-old man underwent the removal of a glioblastoma multiforme from the right parietal lobe of his brain. After the operation, he received x-ray therapy. He made a complete recovery. Today, over 25 years later, he is alive and well and is regularly employed full-time. If glioblastomas and other cerebral gliomas are removed completely, the patients can be cured of their tumors. PMID- 2987749 TI - Influence on the emotional reinforcing systems of the brain as a method of pathogenetic therapy of alcoholism and toxicomania. PMID- 2987750 TI - Experimental encephalitis in monkeys caused by the Powassan virus. AB - We have carried out a comparative study of the experimental infection of monkeys with the P-40 strain of the Powassen virus, isolated in the Primor'e Territory of the USSR, and with the Canadian prototype LB strain. The Powassan virus was found to be pathogenic for Macaca rhesus. The clinical and pathomorphological picture of the experimental encephalitis was studied, and the full identity of the infection produced in the monkeys by the P-40 strain and the Canadian LB strain of the Powassan virus was demonstrated. On electron microscopic examination of the central nervous system the virus was detected in the neurons, glial cells, and intercellular spaces. The virions of the strains studied have identical morphological parameters, being 37-45 nm in diameter and of spherical shape. The data obtained indicated a marked neurotropism of the virus. They will contribute to the elucidation of the role of the virus in the infection pathology of humans, i.e., in the differentiation of encephalitis cases not associated etiologically with the virus of the spring-summer tickborne encephalitis. PMID- 2987752 TI - Simulation of hippocampal afterdischarges synchronized by electrical interactions. AB - Recent experiments have shown that hippocampal pyramidal cells can generate synchronized action potentials even when chemical synapses are blocked. The computer simulations reported here showed that communication between cells by extracellular currents could cause this synchrony, provided that (1) individual neurons were sufficiently excitable and that (2) the resistivity of the extracellular medium was sufficiently high. Synchronization was enhanced if electronic junctions were also present. PMID- 2987751 TI - Mechanisms of the central action of lithium. PMID- 2987753 TI - Oscillations of the spontaneous slow-wave sleep rhythm in lateral geniculate nucleus relay neurons of behaving cats. AB - Intracellular recordings of 31 lateral geniculate nucleus relay neurons were performed in darkness in behaving cats in order to analyse electrical postsynaptic events which appeared during slow-wave sleep. A specific pattern characterized slow-wave sleep: a rapid depolarizing potential arising from baseline initiated a slow depolarization lasting for 40-60 ms which in turn most often elicited delayed fast spikes. This pattern recurred at a frequency of 6 12/s. The slow depolarizations were voltage dependent, usually not separated by any obvious phasic hyperpolarization and showed refractoriness. Other rapid depolarizing potentials occurring during the time course or at the end of a slow depolarization could have generated spike(s) but were followed by a rapid decay. Slow depolarizations were not observed during arousal or paradoxical sleep when the neurons tonically depolarized and displayed either rapid depolarizing potentials with a fast decay or repetitive firing and long high frequency bursts. In five of the studied neurons, decreases in frequency of the spontaneous rapid depolarizing potentials occurred during slow-wave sleep for 3-30 s oscillatory periods without any change in the behavioural state. During these periods all of the few remaining rapid depolarizing potentials arose from a flat baseline, had a higher amplitude and initiated a slow depolarization which always elicited a spike or burst of spikes after a brief delay. The slow-wave sleep rhythm decreased to 1-5/s. Simultaneously the baseline membrane potential hyperpolarized by a few millivolts and reached a level for reversal of inhibitory postsynaptic potentials. Imposed hyperpolarization of the membrane during wakefulness did not reveal any slow depolarization. But strong synaptic excitatory inputs and direct excitation (a break of the current pulse) from a hyperpolarized membrane did evoke the slow depolarization and eventually the fast spike(s) in both control and oscillatory neurons. A rhythm similar to that of slow-wave sleep was elicited during wakefulness by optic tract stimulation and was enhanced by membrane hyperpolarization. But under these conditions the rhythm was initiated by a phasic hyperpolarization and was composed of an alternating hyperpolarization depolarization. Spontaneously and synaptically evoked rapid depolarizing potentials arising from baseline had a similar rising slope. The spontaneous ones initiated a slow depolarization leading to fast spike(s) during slow-wave sleep and could directly generate fast spike(s) during wakefulness.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2987754 TI - On the location of gamma-aminobutyrate and benzodiazepine receptors in the cerebellum of the normal C3H and Lurcher mutant mouse. AB - Binding of gamma-aminobutyrate and benzodiazepine receptor ligands has been studied in the cerebellum of adult normal (C3H) and Lurcher mutant mice. The adult mutant has lost all Purkinje cells and more than 90% of the granule cells in the cerebellar cortex. When compared with their normal littermates Lurcher mice displayed large decreases in the number of high-affinity binding sites for [3H]muscimol, a synaptic gamma-aminobutyrate receptor ligand, in washed cerebellar homogenates. This observation was consistent with the extensive loss of gamma-aminobutyrate receptive Purkinje and granule cells from the Lurcher cerebellum. However, specific binding of the benzodiazepine-receptor ligand [3H]flunitrazepam to Lurcher cerebellum remained unchanged. Indeed quantitative autoradiography, employing [3H]flunitrazepam as a photoaffinity label, showed no significant differences in the density of labelling between Lurcher and normal littermate mice in any region of the cerebellum. These benzodiazepine binding sites in washed homogenates or tissue sections displayed a gamma-aminobutyrate induced enhancement of [3H]flunitrazepam binding which occurred to the same extent in both Lurcher and normal cerebellum, a facilitatory effect which could be blocked by the addition of bicuculline methobromide. Our results suggest that a large proportion of the high-affinity, specific benzodiazepine binding sites in mouse cerebellum are not coupled to the synaptic gamma-aminobutyrate receptors thought to be labelled by high affinity [3H]muscimol binding. Further, that benzodiazepine binding sites do not appear to be enriched on either the soma or dendrites of Purkinje cells, as has been suggested from previous studies. Investigations at the electron microscope level are now required to elucidate the cellular location of benzodiazepine binding sites in the cerebellar cortex and to examine whether or not they are likely to be exposed to gamma-aminobutyrate in vivo. PMID- 2987755 TI - A serotonin-containing pathway from the area postrema to the parabrachial nucleus in the rat. AB - A combined retrograde tracing-immunofluorescent technique was used to identify the relationships between the cellular population projecting to the parabrachial nucleus and the serotonin-immunoreactive cell population of the area postrema in rats. The retrograde fluorescent tracer True Blue was injected in the parabrachial region and 3 days later the animals were perfused. Serial cryostat sections were processed for serotonin immunofluorescence. Three different groups of labeled cells were identified in the area postrema. First, True Blue-positive cells (up to 250/section) that project to the parabrachial nucleus were observed distributed throughout the area postrema. Second, in the pargyline (a monoamine oxidase inhibitor)-treated animals a large number of serotonergic cells (up to 125/section) was observed distributed throughout the area postrema. There was a tendency to a heavier distribution of serotonin-immunoreactive cells in the dorsal two-thirds of the area postrema. Third, double-labelled cells were also seen. Twenty percent of the True Blue-labelled cells projecting to the parabrachial nucleus were serotonin-immunoreactive. Thirty nine percent of the serotonin-immunoreactive population was retrogradely labelled with True Blue. Thus a new serotonergic pathway from the area postrema to the parabrachial nucleus is described; this pathway may be important in the ascending transmission and modulation of chemical and visceral sensory input. PMID- 2987756 TI - alpha 1- and beta-adrenergic receptors are co-regulated during both noradrenergic denervation and hyperinnervation. AB - Changes in the density of alpha 1- and beta-adrenergic receptors were studied following denervation of rat cerebral cortex and hyperinnervation of cerebellum and motor trigeminal nucleus, caused by neonatal 6-hydroxydopamine treatment. Four well-defined thalamic projection zones to cortex were studied separately using tissue punch methodology. Both alpha 1- and beta-adrenergic receptors were unevenly distributed in motor, sensory, visual and auditory cortex. The density of alpha 1-adrenergic receptors correlated better with the norepinephrine content of the punches (r = 0.62) than did the density of beta-adrenergic receptors (r = 0.38). Noradrenergic denervation increased both alpha 1- and beta-adrenergic receptor density in almost all cortical areas studied, however the percentage increase was larger for beta- than alpha 1-adrenergic receptors. The change in receptor density was largest in visual cortex and smallest in somatosensory cortex for both receptor sub-types. Noradrenergic hyperinnervation caused a 15 18% decrease in both alpha 1- and beta-adrenergic receptor density in the motor trigeminal nucleus of the pons, but did not change the density of either receptor type in the cerebellum. In general, following either noradrenergic denervation or hyperinnervation the change in alpha 1-adrenergic receptor density was correlated (r = 0.64, P less than 0.005) with the change in beta-adrenergic receptor density in each region, suggesting that these different receptor sub-types are under similar control mechanisms. PMID- 2987758 TI - Physiologic assessment of phosphoglycerate mutase deficiency: incremental exercise test. AB - A third case of phosphoglycerate mutase (PGAM) deficiency, a metabolic myopathy involving terminal glycolysis, was identified in a 24-year-old black man with episodic, exercise-induced myoglobinuria since age 13. To better understand the physiologic consequences of PGAM deficiency, incremental exercise testing was performed. Results were compared with those of two patients having myophosphorylase deficiency and five normals. In contrast to the patients with phosphorylase deficiency, the PGAM-deficient patient achieved near-normal levels of maximal exercise and produced a normal peak lactate after exercise. The mechanisms underlying the asymptomatic performance of such strenuous exercise in this case are uncertain, but the data suggest that unidentified factors are operative in precipitating attacks of myoglobinuria in patients with some metabolic myopathies. Despite similar clinical histories, patients with different glycolytic enzyme deficiencies can have striking differences in exercise tolerance. PMID- 2987757 TI - Fatal infantile cytochrome c oxidase deficiency: decrease of immunologically detectable enzyme in muscle. AB - A 2-month-old boy had progressive generalized weakness, hypotonia, and respiratory insufficiency requiring assisted ventilation. At age 3 1/2 months, he started having seizures and recurrent pulmonary infections; he died at age 7 months. Serum lactate was chronically elevated, but there was no aminoaciduria. Histochemical and ultrastructural studies of muscle biopsies at ages 2 and 3 months showed excessive mitochondria, lipid, and glycogen; a third biopsy at 6 months showed marked increase in perimysial fibrous and fat tissue. Cytochrome c oxidase activity was 7% of normal in the first biopsy and undetectable in the others. Cytochrome spectra of mitochondria isolated from postmortem muscle showed complete lack of cytochrome aa3. Antibodies were obtained against cytochrome c oxidase purified from normal human heart. Immunotitration and enzyme-linked immunosorbent assay (ELISA) showed decreased immunologically reactive enzyme protein in the patient's muscle, but SDS-PAGE electrophoresis of immunoprecipitates of muscle mitochondrial extracts showed the presence of all cytochrome c oxidase subunits. These data suggest that decreased synthesis of one or more subunits may result in markedly decreased concentration of electrophoretically normal complex IV in skeletal muscle. PMID- 2987759 TI - [Multiple carcinomas of the large intestine]. PMID- 2987760 TI - [Prognostic significance of plasmatic concentration of fibronectin in patients with bronchial carcinoma]. AB - Plasma levels of fibronectin were determined in 50 lung cancer patients and in 25 normal subjects. Immunoturbidimetric assay was performed to detect possible variations of plasma fibronectin concentration in neoplastic patients. Moreover plasma levels of fibronectin were related to clinical stage, histological type of tumor and survival after surgery. PMID- 2987761 TI - Membrane responses and changes in cAMP levels in Aplysia sensory neurons produced by serotonin, tryptamine, FMRFamide and small cardioactive peptideB (SCPB). AB - While recent evidence indicates a role for serotonin (5-HT) in modulating the defensive tail-withdrawal reflex in Aplysia, little information exists concerning the specificity of these 5-HT effects. As a first-step in addressing this issue we have examined the dose-response relationship for one aspect of the 5-HT modulation (enhancement of cAMP levels in isolated clusters of sensory neurons) and compared the effects of 5-HT with three potential neurotransmitters: tryptamine, FMRFamide (Phe-Met-Arg-Phe-NH2) and small cardioactive peptideB (SCPB). Cyclic adenosine monophosphate (cAMP) levels were enhanced as a graded function of the concentration of 5-HT with an EC50 of 14 microM. At a concentration of 5 microM, both 5-HT and SCPB produced nearly identical increases in the cAMP content of sensory neurons. In contrast, 5 microM tryptamine or 5 microM FMRFamide had little or no effect on cAMP levels. We also examined the effects of these agents on membrane currents and membrane conductance. Both 5-HT and SCPB produced an inward current associated with a decrease in input conductance. Tryptamine had little or no effect, while FMRFamide produced a response opposite to that of 5-HT and SCPB; an outward current associated with an increase in membrane conductance. PMID- 2987762 TI - Astrocyte opioid receptors: activation modifies the noradrenaline-evoked increase in 2-[14C]deoxyglucose incorporation into glycogen. AB - Astrocyte-enriched cultures of the newborn rat cortex accumulated 2 [14C]deoxyglucose (2-DG), a proportion portion of which was incorporated into glycogen. Exposure to exogenous noradrenaline resulted in an increase (30%) in the incorporation of 2-[14C]DG into glycogen in these cultures. The extent to which noradrenaline evoked an increase in 2-[14C]DG labelling of glycogen was modified by both morphine and methionine-enkephalin. Whereas morphine augmented the noradrenaline-induced increase in 2-[14C]DG incorporation into glycogen in these cultures, methionine-enkephalin attenuated this response. PMID- 2987763 TI - Autoreceptors regulate gamma-[3H]aminobutyric acid release from the guinea pig small intestine. AB - The possible presence of a presynaptic gamma-aminobutyric acid (GABA) receptor capable of regulating the release of GABA was investigated using the guinea pig small intestine. Muscimol at 10(-8) M and 10(-7) M, but not baclofen at 10(-6) M, inhibited the K+ (40 mM)-evoked Ca2+-dependent release of [3H]GABA from the small intestine preloaded with [3H]GABA, in the presence of 10(-6) M tetrodotoxin. The effect of muscimol on the K+-evoked GABA release was inhibited by bicuculline and furosemide. These results show that the guinea pig small intestine possesses presynaptic GABA receptors which may be involved in the regulation of the evoked GABA release. The presynaptic GABA receptor is bicuculline-sensitive and is probably coupled to the Cl- ion channel. PMID- 2987764 TI - Cellular uptake of cobalamin. PMID- 2987765 TI - The radionuclide evaluation of septal wall motion following coronary bypass surgery. AB - We investigated the diagnostic accuracy of radionuclide global and regional left ventricular (LV) function after coronary revascularization. A consecutive series of 43 patients was studied. First-pass radionuclide angiograms were performed preoperatively (4 days +/- s.d. 3 days, range 1-18 days) and postoperatively (7 days +/- s.d. 3 days, range 3-19 days). Regional radionuclide LV function was assessed using the two-dimensional display of systole and the three-dimensional ejection fraction image. Electrocardiograms were obtained the day prior to surgery and every 8 h for the first three postoperative days. Creatine kinase (CK) and lactate dehydrogenase (LDH) isoenzymes were obtained the day prior to surgery and every 8 h for the first three postoperative days. In 39 patients who did not develop perioperative myocardial infarction by isoenzyme or ECG criteria, we found that 38 patients showed unchanged or improved global and regional LV function, while one patient without isoenzyme or ECG evidence of perioperative myocardial infarction developed a new septal wall motion abnormality. Thus, the specificity of the radionuclide radioventriculogram for new septal wall motion abnormalities was very high. In four patients who developed isoenzyme and ECG evidence of myocardial infarction, septal wall motion worsened in all four patients while global left ventricular ejection function fell significantly in three patients. Thus, the radionuclide radioventriculogram also had high diagnostic sensitivity. In summary, contrary to past and recent reports, this investigation demonstrated that the radionuclide radioventriculogram can be used to assess global and regional LV function after coronary artery bypass surgery and furthermore, that it reliably indicates the presence of a new postoperative myocardial infarction. PMID- 2987766 TI - Treatment of high-risk gestational trophoblastic disease with methotrexate, actinomycin D, and cyclophosphamide chemotherapy. AB - Seventy-three patients with metastatic high-risk gestational trophoblastic disease were treated with methotrexate, actinomycin D, and cyclophosphamide chemotherapy at the Brewer Trophoblastic Disease Center between 1968 and 1982. Forty-six patients were treated primarily with methotrexate, actinomycin D, and cyclophosphamide because of the presence of one or more high-risk factors. Twenty seven additional patients who had not responded to initial single-agent chemotherapy with methotrexate and/or actinomycin D were subsequently treated with methotrexate, actinomycin D, and cyclophosphamide. Adjuvant surgery and radiotherapy were used in selected patients. The overall cure rate was 51% (37 of 73): 63% (29 of 46) for primary treatment and 30% (eight of 27) for secondary treatment (P less than .01). Several factors that influenced response to primary treatment with methotrexate, actinomycin D, and cyclophosphamide chemotherapy were determined: 1) clinicopathologic diagnosis of choriocarcinoma versus invasive mole (59 versus 100%), 2) metastases to sites other than the lung and/or vagina (44 versus 74%), 3) antecedent term gestation compared with hydatidiform mole or abortion (50 versus 75%), and 4) presence of three or more high-risk factors (27 versus 74%). There were no significant differences in cure rates during the course of the study period. PMID- 2987767 TI - [Obesity therapy--sensible after care of patients following treatment using a nutrition counseling system]. PMID- 2987768 TI - [Tissue polypeptide antigen (TPA) and plasma TPA levels during the postoperative course in female primary breast carcinoma patients. 1st results of a long-term study]. AB - TPA determinations can be used for monitoring advanced cancer patients. Increasing and decreasing TPA levels can be correlated to the further course of the disease. TPA levels should be determined in the postoperative follow-up of cancer patients with a higher risk of recurrence. Single point determinations for primary diagnosis is not recommended. Only the interpretation of long term evaluation studies in combination with clinical findings can be an advantage in managing cancer patients. PMID- 2987769 TI - Acquired immune deficiency syndrome. Pathogenic mechanisms of ocular disease. AB - A gross, light, and electron microscopic study of the eyes from 35 consecutive autopsy cases of the acquired immune deficiency syndrome revealed cotton-wool spots (71% of cases), retinal hemorrhage in areas without cytomegalovirus infection (40%), cytomegalovirus retinitis (34%) with associated retinal detachment, Roth's spots (23%), retinal microaneurysms (20%), papilledema (14%), conjunctival Kaposi's sarcoma (9%), cryptococcal chorioretinitis (6%), Mycobacterium avium-intracellulare in retina and in choroidal granulomas (6%), ischemic maculopathy (6%), bilateral keratitis (3%), and herpes simplex retinitis (3%). Ocular infection with candida or toxoplasmosis were not found in this autopsy series. Immunocytologic studies demonstrated deposition of immunoglobulins in arteriolar walls, consistent with immune complex mediated disease. Ultrastructural studies showed a vasculopathy in the areas near cotton wool spots. A mechanism is proposed linking the deposition of immune complexes with subsequent small vessel lesions, ischemia, cotton-wool spots and later spread of cytomegalovirus to retina via damaged vascular endothelium. PMID- 2987771 TI - [Clinical aspects of the narrow spinal canal]. AB - Not infrequently the lumbosacral root-compression syndrome is due to stenosis of the lumbal spinal canal. The clinical symptoms, usually emerging in the middle aged, may be characterized by either simple radicular pain with or without neurological deficit or by exercise-dependent transient functional disturbance of the cauda equina, namely, the so-called neurogenic intermittent claudication (NIC). In most cases, NIC is due to mechanical compression of the cauda fibers as a result of extension of the patient's lumbar spine. In some patients, exercise induced ischemic radiculopathy is assumed to be the main factor. Differential diagnosis must take into account true intermittent claudication, certain rare myopathies (if the exercise-dependent pain is the main feature), and certain transient disturbances in spinal cord function (if a motor and/or sensory deficit arises during the NIC attack). PMID- 2987770 TI - Synthetics in anterior cruciate ligament reconstruction: a review. AB - Because of a rapidly growing interest in synthetic ligaments, the American Academy of Orthopedic Surgeons Committee on the Knee sponsored a seminar in Atlanta, Georgia, in June 1983. Attending this seminar were Knee Committee Members, six invited guest consultants with expertise in collagen metabolism and healing and materials testings, and eight teams of investigators who presented data on eight different devices. In November 1983, the Federal Food and Drug Administration Advisory Panel for Orthopedics held an open meeting in Washington, D.C., on the subject of artificial ligaments. In February 1984, there was an hour and a half symposium on artificial ligaments at the Annual Meeting of the American Academy of Orthopedic Surgeons. Although there is widespread interest in this subject, it is evident that more scientific information and clinical experience must be accumulated. Uniform methods of evaluation, both biomechanical and clinical, are needed. At present, synthetic materials are being investigated under rigid controls, and none have been released for general clinical use. Indications are limited to cases of chronic instability in which autogenous tissues are insufficient or unusable. PMID- 2987772 TI - [Results of treatment of intradisk therapy with collagenase in lumbar intervertebral disk prolapse]. PMID- 2987773 TI - ACTH-dependent modulation of malaria immunity in mice. AB - Tetracosactrin, a synthetic adrenocorticotrophic hormone (ACTH) analogue delivered by osmotic minipumps implanted s.c. in mice induced a dose-dependent increase of plasma corticosterone levels. In mice with an established immunity to Plasmodium berghei the increase of the plasma corticosterone level due to tetracosactrin treatment correlated with loss of immunity against this malaria parasite. The observed plasma corticosterone levels associated with loss of malaria immunity were of the same order as those in mice that lost their immunity during pregnancy. Adrenalectomy before administration of the ACTH analogue prevented both the increase of plasma corticosterone and loss of malaria immunity. Adrenalectomized mice still lost their malaria immunity when treated with the synthetic corticoid dexamethasone. The effector function of malaria immunity is sensitive to corticoids, and, at least during pregnancy, the naturally occurring serum corticosterone level appears to be an important regulator of malaria immunity. PMID- 2987774 TI - Rotavirus electrophoretypes in infantile gastro-enteritis. An epidemiological study. AB - From January first 1978 to September 30, 1981, 93 fecal samples from children hospitalized with a rotavirus gastro-enteritis were tested after RNA extraction and polyacrylamide gel electrophoresis. 14 electrophoretypes (TI--T14) were identified by this technique. They were studied in relation with epidemiological as well as clinical symptoms. No electrophoretype correlated well with age or living area. The electrophoretype T3 was found only in male infants and the electrophoretype T7 was 3 times more frequent in boys than in girls. The electrophoretype T4 was prevailing during the winter 1980-1981 and the electrophoretype T5 prevailed during the summer 1981. 19 infants were probably contaminated during their stay in the hospital, 9 of them with the electrophoretype T4 prevailing at that time. The prevalence of associated otitis and rash was significantly higher with the electrophoretype T3. The polyacrylamide gel electrophoresis technique appears to be a simple, and reliable epidemiological tool to evaluate rotavirus infection in communities. PMID- 2987775 TI - Sialadenoma papilliferum. AB - Sialadenoma papilliferum is a rare benign papillary intraoral tumour most frequently affecting men over the age of 50. The tumour bears many similarities to adenolymphoma of the parotid and may be considered a variant of the latter. A case is presented and the literature is reviewed. PMID- 2987776 TI - Electron microscopy after direct ultracentrifugation. AB - A direct ultracentrifugation technique was used in the preparation of skin lesion specimens for examination by electron microscopy. The concentration factor of centrifuged specimens was estimated to be in excess of 1,000-fold compared to conventional adsorption techniques. This resulted in an increase of over 300% in the detection rate of herpesviruses and poxviruses from skin lesion specimens. PMID- 2987777 TI - Primary colorectal signet-ring cell carcinoma in young patients: report of 3 cases. AB - Three cases of primary signet-ring cell carcinoma of the rectum are described. They accounted for 0.2% of the 1531 cases of colorectal adenocarcinoma in the 12 yr period from 1972-1983 in the University Department of Pathology at Queen Mary Hospital. The patients were young, aged 18, 24 and 27 yr respectively, in striking contrast to the mean age of 62 in patients with the usual types of colorectal cancer. They were also younger than most patients with this tumour in the literature. They presented with alteration of bowel habit, blood and mucus in stool, and weight loss. Pathological features included constrictive narrowing of the gut lumen by intestinal wall thickened by a desmoplastic reaction to diffusely infiltrating signet-ring carcinoma cells, widespread lymph node and peritoneal metastases, and absent hepatic metastasis. Microscopically, the mucosa was largely intact, but had multifocal tumour involvement. This peculiar feature was responsible for three consecutive negative biopsies in one case. Care in distinguishing it from mucinous adenocarcinoma is emphasized. All three patients presented with Dukes' C lesions. The prognosis is poor. PMID- 2987778 TI - Primary malignant fibrous histiocytoma of the uterus--ultrastructural and immunocytochemical studies of two cases. AB - The clinical and pathological features, including ultrastructural and immunocytochemical findings, of 2 cases of primary malignant fibrous histiocytoma of the uterus are reported. One patient had pulmonary metastases at the time of hysterectomy: she developed pelvic recurrence and died 3 mth after operation. The other patient was well and apparently free of disease 6 mth after hysterectomy. Histologically both tumours resembled pleomorphic leiomyosarcoma, and the diagnosis of a malignant fibrous histiocytoma was confirmed only after electron microscopic and immunocytochemical studies. Hormone receptor assay for estrogen and progesterone was positive in both tumours. Anti-estrogen treatment was started in one case but was ineffective. PMID- 2987779 TI - The functional stability of the nervous system: a neurobiological basis of intelligence? PMID- 2987780 TI - Acute hepatitis--A, B, NANB and delta. PMID- 2987781 TI - The diagnosis and management of Wilms' tumor. AB - Wilms' tumor is the most frequent malignant tumor of the kidney in infants and children. Effective treatment requires abdominal exploration with nephrectomy and administration of postoperative combination chemotherapy, with abdominal irradiation employed only for the treatment of microscopic or gross residual abdominal disease. Current therapeutic research is focused on improving therapy for certain patient groups, such as those with sarcomatous tumors, who continue to have a poor prognosis, and designing less toxic treatment programs for those patients who currently have a favorable prognosis. PMID- 2987782 TI - Characterization of HCO3-/CO2 pool sizes and kinetics in infants. AB - The first bicarbonate pool sizes and kinetic data necessary for the interpretation of substrate oxidation studies have been determined in six fed, nonacidotic infants, ages 2.5 to 5 months. Following an intravenous bolus of NaH13CO3 (50 mumol/kg), breath samples were collected over 240 min for the analysis of breath 13CO2. Each breath 13CO2 disappearance curve was fitted to a multicompartmental bicarbonate model previously derived in adults. The mean sizes of the three bicarbonate pools were: 7.4 +/- 0.8 mmol/kg (central pool), 15.1 +/- 4.8 mmol/kg (rapidly exchanging peripheral pool), and 8.8 +/- 3.5 mmol/kg (slowly exchanging peripheral pool). The mean percentage dose recovery was 57 +/- 10%. The pool sizes suggested that extensive metabolic exchange of carbon between HCO3 and organic metabolites occurred in the infant. PMID- 2987783 TI - Clinical-histopathological correlations in experimental otitis media: implications for silent otitis media in humans. AB - Clinical-histopathological correlations were sought in an experimental animal model of otitis media. Among 20 chinchillas inoculated intranasally with wildtype influenza A/Alaska virus (H3N2) and type 7F Streptococcus pneumoniae, 15 animals (18 ears) developed otoscopic and tympanometric signs of otitis media with middle ear effusion. Middle ear inflammation was most intense 10 days after virus inoculation. Twenty-two days after virus inoculation, eight ears showed diffuse middle ear histopathology and contained effusion, which cultured pneumococcus, five ears showed focal histopathology, and four of the five contained no effusion. Clinical manifestations of otitis media had disappeared in four of the 13 ears with pathology at sacrifice; otoscopy was normal in all four ears, tympanometry was normal in three ears, and both otoscopy and tympanometry were normal in one ear. All four of these ears with clinically "silent" middle ear histopathology had shown abnormalities of ear drum appearance or tympanometry between 7 and 14 days after inoculation. Discordance between histopathology and the clinical examination performed at sacrifice was greatest for ears with focal pathology. PMID- 2987784 TI - Epstein-Barr virus infectious mononucleosis in children. I. Clinical and general laboratory findings. AB - Between 1976 and 1982, 113 children aged 6 months to 16 years with documented Epstein-Barr virus-induced infectious mononucleosis were studied prospectively, and in most instances serially. An unexpected finding was the large number of young children, less than 4 years old, with this disease. Children with infectious mononucleosis, in particular the very young, tended to have more rashes, significant neutropenia, abdominal pain (older children only), and possible hepatosplenomegaly than have been reported in adult patients. The intensity of the characteristic relative atypical lymphocytosis found in peripheral blood was age-related; it was less in the very young. Findings of failure to thrive, otitis media, and episodes of recurrent tonsillopharyngitis appeared to be unique or more closely associated with childhood disease. Complications such as thrombocytopenia with hemorrhagic manifestations, significant airway obstruction, and neurologic problems occurred more frequently whereas jaundice occurred less frequently than noted in adult patients. Six children, all less than 4 years old, developed pneumonia during the disease course. The increased availability of Epstein-Barr virus-specific testing should continue to expand our knowledge of this disease in children of all ages. PMID- 2987785 TI - Epstein-Barr virus infectious mononucleosis in children. II. Heterophil antibody and viral-specific responses. AB - An investigation was performed to address the need to establish the rate of positive heterophil antibody responses, oropharyngeal isolation of Epstein-Barr virus (EBV), and the evolving pattern of EBV-specific antibody responses among children with documented EBV-infectious mononucleosis. Findings showed that the rate of heterophil antibody responses appeared to increase progressively with advancing age from infancy up to 4 years, after which the rates approached values similar to that reported in young adult patients. The rapid slide test detected a heterophil antibody response as frequently as the Paul-Bunnell-Davidsohn horse cell test, except in children less than 4 years old. The decreased sensitivity found with the rapid slide test in the very young was associated with their less intense heterophil response. The younger group of children also developed a lower acute mean titer and, as a result, a decreased persistence of immunoglobulin M antibody to EBV-capsid antigen, whereas they had more frequent responses to EBV early antigen directed to restricted component than both the older subjects and adults reported elsewhere. Antibodies to EBV-nuclear antigen, characteristically a late-onset antibody, tended to develop earlier than noted in adult patients. In contrast, the prevalence and continued excretion of EBV from oropharyngeal secretions was similar to that reported in adult patients. It is speculated that these age-related differences in host responses are associated with the ontogeny of the immunologic system. PMID- 2987786 TI - Sudden hearing loss following infectious mononucleosis: possible effect of altered immunoregulation. AB - Sudden, permanent hearing loss developed in three young patients. In an attempt to determine a viral etiology, their humoral and cell-mediated responses to a panel of seven viruses linked to deafness and altered immunity were measured. Although a specific viral cause was not determined, a mild Epstein-Barr virus infection was documented for each patient. It had preceded their hearing loss onset by 1 to 4 months. Evidence of altered cell-mediated responses to Epstein Barr virus antigens was found in each patient. It is proposed that, in certain susceptible individuals, a temporary cellular immunosuppression, which accompanies normal recovery from Epstein-Barr virus infection, may provide an opportunity for a viral invasion of, or a latent viral reactivation in, the inner ear. This invasion or reactivation leads to deafness. PMID- 2987787 TI - [Circulating immune complexes in adenovirus infections in young children]. PMID- 2987788 TI - [Congenital cytomegalic infection manifesting itself as a severe nephrotic syndrome simulating microcystosis of the kidneys (nephrotic syndrome of the Finnish type)]. PMID- 2987789 TI - A comparative study of the interaction of 5,10,15,20-tetrakis (N-methylpyridinium 4-yl)porphyrin and its zinc complex with DNA using fluorescence spectroscopy and topoisomerisation. AB - Binding of 5,10,15,20-tetrakis (N-methylpyridinium-4-yl)porphyrin (H2TMPyP4+) and its zinc complex (ZnTMPyP4+) to DNA is demonstrated by their coelectrophoresis and by absorption and fluorescence spectroscopic methods. Topoisomerisation of pBR322 DNA shows that H2TMPyP4+ unwinds DNA as efficiently as ethidium bromide showing that it intercalates at many sites. ZnTMPyP4+ may cause limited unwinding. Marked changes in the fluorescence spectra of the porphyrins are found in the presence of DNA. The fluorescence intensity of either H2TMPyP4+ or ZnTMPyP4+ is enhanced in the presence of poly (d(A-T)), whereas in the presence of poly (d(G-C] the fluorescence intensity of ZnTMPyP4+ is only slightly affected and that of H2TMPyP4+ markedly reduced. Both the porphyrins photosensitize the cleavage of DNA in aerated solution upon visible light irradiation. PMID- 2987790 TI - Unique sequences are interspersed among tandemly repeated elements in the murine gamma 1 switch segment. AB - Early in its differentiative pathway, a given B lymphocyte expresses immunoglobulin of the mu heavy chain class (IgM). Subsequent differentiative processes may involve rearrangement within the immunoglobulin heavy chain chromosomal locus to enable cells of the same lineage to synthesize immunoglobulins of other heavy chain classes (e. g. IgG, IgE or IgA), but with specificity for the same antigen as the original IgM molecule. Switch recombination, the molecular event which facilitates this chromosomal rearrangement, has been shown to occur between segments of DNA consisting of tandemly repeated unit sequences. These DNA segments have been functionally defined as switch regions. We have cloned the gamma 1 switch region from the BALB/c germline, and have demonstrated that significantly divergent sequence elements are interspersed among the tandemly repeated units characteristic of this switch region. We show that these unique elements exist in at least three copies within the switch segment, and discuss the implications of this novel and previously unreported primary structure. PMID- 2987791 TI - DNA polymerase alpha and models for proofreading. AB - Using a modified system to measure fidelity at an amber site in phi X174, we have employed DNA polymerase alpha to test different mechanisms for proofreading. DNA polymerase alpha does not exhibit the characteristics of "kinetic proofreading" seen with procaryotic polymerases. Polymerase alpha shows no evidence for a "next nucleotide" effect, and added deoxynucleoside monophosphates do not alter fidelity. Pyrophosphate, which increases error rates with a procaryotic polymerase, appears to weakly improve polymerase alpha fidelity. DNA polymerase alpha does exhibit a dramatic increase in error rate in the presence of a deoxycytidine thiotriphosphate (dCTP alpha S), but this enhanced mutagenesis also occurs under conditions where kinetic proofreading should be otherwise defeated. This particular effect with dCTP alpha S appears specific for DNA polymerase alpha and is not seen with the other polymerases tested. PMID- 2987792 TI - Nucleotide sequences of murine intracisternal A-particle gene LTRs have extensive variability within the R region. AB - Nucleotide sequences of the long terminal repeats (LTRs) of four murine intracisternal A-particle (IAP) genes IAP62, 19, 81 and 14 were determined. Each IAP LTR contains three sequence domains, 5'-U3-R-U5-3', and each is bound by 4 bp imperfect inverted repeats. The transcriptional regulatory sequences, CAAT and TATA, as well as the enhancer core sequence GTGGTAA are conserved and precisely positioned within the U3 region. In the R region, the sequence AATAAA is located twenty base pairs preceding the dinucleotide CA, the polyadenylation site. In IAP19 and IAP81, the 5' and 3' LTRs are flanked by a six nucleotide direct repeat of cellular sequences representing the possible integration sites for these IAP proviruses. Both the size and sequences of different IAP LTRs vary considerably, with the majority of the variation localized within the R regions. The size of R varies from 66 bp in IAP14 to 222 bp in IAP62; in contrast, the U3 and U5 regions are all similar in size. These extra sequences within the R region of large LTRs consist of several unusual directly repeating sequences which account for this variability. PMID- 2987793 TI - Characterization of a factor that can prevent random transcription of cloned rDNA and its probable relationship to poly(ADP-ribose) polymerase. AB - A factor which eliminated nonspecific transcription of cloned rat rDNA was extensively purified from rat mammary adenocarcinoma ascites cells by successive fractionations on DEAE-Sephadex and heparin-Sepharose columns. The fractions containing RNA polymerase I (HS-B) and fractions eluting thereafter (HS-C) from the heparin-Sepharose column were pooled separately. Addition of HS-C to HS-B prevented random transcription of rDNA and yielded an accurate rDNA transcript with negligible non-specific transcription. The factor was essentially homogeneous and corresponded to Poly(ADP-ribose) polymerase with respect to molecular weight, dependence on DNA for its activity and its ability to undergo auto ADP-ribosylation. The total amount of protein in the transcription assay was approximately 2 micrograms, which indicates a high degree of purity of all the factors required for specific transcription of rDNA. PMID- 2987794 TI - The nucleotide sequence recognised by the Escherichia coli D type I restriction and modification enzyme. AB - A type I restriction endonuclease from a new isolate of Escherichia coli (E. coli E166) has been purified and characterised. The enzyme, EcoD, has a recognition sequence similar in overall structure to the previously determined type I enzyme sequences, an exception being that it is degenerate. The sequence is 5'-T-T-A-N-N N-N-N-N-N-G-T-C-Y-3' 3'-A-A-T-N-N-N-N-N-N-N-C-A-G-R-5' where Y is a pyrimidine, R is a purine and N can be any nucleotide. The enzyme methylates adenosyl residues in both strands of the DNA that are separated by ten base pairs, suggesting that the enzyme interacts with DNA along one face of the helix making contacts in two successive major grooves. PMID- 2987795 TI - Nucleotide sequence of a macronuclear DNA molecule coding for alpha-tubulin from the ciliate Stylonychia lemnae. Special codon usage: TAA is not a translation termination codon. AB - The gene-sized macronuclear DNA of the hypotrichous ciliate Stylonychia lemnae contains two size classes of DNA molecules (1.85 and 1.73 kbp) coding for alpha tubulin. Each macronucleus contains about 55000 copies of the 1.85 kbp molecules and about 17000 copies of the 1.73 kbp DNA molecules. Five macronuclear molecules of these sequences were cloned and sequenced, one, from the 1.85 kbp size class in its entirety. The 5 sequences fell into two classes suggesting that Stylonychia lemnae contains at least two different alpha-tubulin genes. All 5 clones show the codon TAA in the same nucleotide positions of the coding region. In this position the TAA codon cannot function as a translational stop codon and we suggest that this codon codes for the amino acid glutamine. The nucleotide sequence of the coding region as well as the encoded amino acid sequence is highly conserved compared to alpha-tubulin genes from vertebrates. The noncoding regions show several putative transcription-regulatory sequences as well as sequences presumably functioning as replication origins. PMID- 2987796 TI - Sequence specific damage of DNA induced by reducing sugars. AB - Reducing sugars induced alkali-labile sites in DNA. The DNA reacted with D fructose 6-phosphate or D-fructose in the presence of Cu2+ was cleaved by the treatment with aqueous piperidine at 90 degrees C for 30 min. Alkali-labile sites were induced frequently at the pyrimidine residues, especially at the pyrimidine residues in pyrimidine-purine (5'----3') sequences. Transition metal ions such as Cu2+ and oxygen radicals such as hydrogen peroxide were possibly involved in the induction of alkali-labile sites. PMID- 2987797 TI - Comparison of genomic fragment and clone sequences within a long interspersed repeated sequence of the mouse genome. AB - The 393bp nucleotide sequence of a HindIII genomic fragment mapping within the major long interspersed repeated sequence family (MIF-1, Bam, L1) of mouse is reported and compared to clone sequences of the same region of this repeated sequence. The consensus of the clone sequences significantly differs from the genomic fragment sequence by additions and deletions that are inconsistent with the physical and biochemical properties of the genomic fragment. While alternative explanations could account for some of these differences, several aspects of the experimental results imply that cloning artifacts contribute to the discrepancies. Despite the differences between the clone and genomic fragment sequences, the biologically interesting features previously noted in clone sequences (promoter-like signals and an open reading frame) are conserved in the genomic fragment sequence. PMID- 2987798 TI - Distinct characteristics of loop sequences of two Drosophila foldback transposable elements. AB - A few foldback (FB) transposable elements have, between their long terminal inverted repeats, central loop sequences which have been shown to be different from FB inverted repeat sequences. We have investigated loop sequences from two such FB elements by analyzing their genomic distribution and sequence conservation and, in particular, by determining if they are normally associated with FB elements. One of these FB loop sequences seems to be present in a few conserved copies found adjacent to FB inverted repeat sequences, suggesting that it represents an integral component of some FB elements. The other loop sequence is less well-conserved and not usually associated with FB inverted repeats. This sequence is a member of another family of transposable elements, the HB family, and was found inserted in an FB element only by chance. We compare the complete DNA sequences of two HB elements and examine the ends of four HB elements. PMID- 2987799 TI - Expression of polyoma early gene products in E. coli. AB - The three products of the early region of polyoma virus have been cloned for expression in E. coli using the Tac promoter. Although the identical promoter and ribosome binding site are used in each final construction, the observed level of protein expression is different for each protein. While plasmids expressing wild type T antigens as well as a plasmid expressing the truncated Py-1387T middle T antigen lacking the membrane-anchoring sequence give rise to synthesis of proteins readily detectible by 35S-methionine labeling and immunoprecipitation, only small T and the middle T of Py-1387T are made in amounts sufficient for ready detection in total cell protein. Unlike middle T expressed in animal cells, middle T produced in E. coli is not detectibly phosphorylated. Further, the E. coli protein lacks tyrosine kinase activity. PMID- 2987800 TI - Recent amplification of an alpha satellite DNA in humans. AB - A repeat sequence 682 base pairs (bp) long produced by cleavage of human DNA with Xba I restriction enzyme is composed of four tandemly arranged subunits with lengths of 171, 170, 171, and 170 bp each. The sequence organization of the 682 bp Xba I repeat bears a striking resemblance to other complex satellite DNAs of primates, including the Eco RI human alpha satellite family which also occurs as a 170 bp repeat. The Eco RI tetramer and the 682 bp Xba I repeat show a sequence divergence of 21%. The 682 bp Xba I repeat sequence is restricted to humans and is only distantly related to the previously reported 340 bp Xba human repeated DNA sequence. These finding are consistent with the concept of occasional amplifications of members or groups of members of alpha satellite DNA during human evolution. Amplifications apparently occurred after humans, apes and gibbons diverged from Old World monkeys (Eco RI satellite), after humans and apes diverged from gibbons (340 bp Xba I satellite) and after humans diverged from the great apes (682 bp Xba I satellite). PMID- 2987801 TI - Characterization of a mouse somatic cytochrome c gene and three cytochrome c pseudogenes. AB - Mouse contains two functional, but differentially expressed, cytochrome c genes. One of these genes is expressed in all somatic tissues so far examined. The other gene is expressed only in testis and is assumed to be spermatogenesis-specific. The nucleotide sequence of four mouse cytochrome c-like genes has been determined. One of these genes (MC1) contains an intron and encodes a polypeptide sequence identical to the published mouse somatic cytochrome c amino acid sequence. The other three genes can not properly encode a mouse cytochrome c protein and appear to be pseudogenes which have arisen via an insertion into the mouse genome of a cDNA copy of a cytochrome c mRNA molecule. PMID- 2987803 TI - A novel strategy for constructing clustered point mutations. AB - We have modified the synthetic linker mutagenesis procedure (1,2) in order to facilitate both the construction and the analysis of deletions, insertions and clustered point mutations generated in DNA in vitro. The protocol as originally described by McKnight and Kingsbury (1) involved attaching a synthetic linker sequence to each 5' or 3' deletion endpoint in DNA. We have designed specific plasmid vectors that can be used to generate nested sets of deletion mutations in the DNA being analyzed. The utility of these vectors is that a linker sequence of choice can be inserted at the endpoint of a deletion in a single intramolecular ligation without the use of synthetic linker DNA. In a second modification of the original procedure, we have adopted a rapid method for sequencing supercoiled plasmid DNAs from 10 ml cultures by primer extension. The site-directed mutagenesis strategy outlined here is suited for studying regulatory regions of DNA, such as origins of DNA replication, transcriptional promoters, enhancer elements, and activator binding sites. We have used this rapid and efficient strategy to generate deletions, insertions, and clustered point mutations in the transcriptional control region of a gene encoding the major human ribosomal RNAs. PMID- 2987802 TI - Characterization of two Drosophila melanogaster cytochrome c genes and their transcripts. AB - Analysis of total Drosophila melanogaster DNA by genomic blot hybridization indicates that two cytochrome c-like sequences exist in the Drosophila genome. These two sequences, DC3 and DC4, have been isolated from a Charon 4A-D. melanogaster genomic library. DC3 and DC4 are located within a 4 kb region of DNA, at position 36A 10-11, on the left arm of chromosome 2. The nucleotide sequence of these two clones has been determined. Both DC3 and DC4 can encode functional cytochrome c proteins. The polypeptide sequences predicted by these two genes, however, differ at 32 amino acid residues. DC4 is expressed at varying, but relatively high levels throughout Drosophila development. In contrast, DC3 is expressed at constant, but relatively low levels throughout development. PMID- 2987804 TI - Sequence organisation of rat seminal vesicle F gene: location of transcriptional start point and sequence comparison with six other androgen-regulated genes. AB - Seminal vesicle F gene, encoding an androgen-regulated serine-rich structural protein of the rat copulatory plug, has been sequenced together with 5' and 3' flanking regions. The intron/exon arrangement of the gene deduced from restriction maps was confirmed. The major and possible minor transcriptional start points were located by primer extension analysis and S1 nuclease mapping. A published nucleotide sequence for seminal vesicle S gene which also encodes an androgen-regulated protein of the copulatory plug has been extended to allow comparison of F and S genes. The considerable sequence homology between the two genes confirms their evolutionary relatedness. Homology is especially high in their promoter regions and their transcriptional start points are identical. They share several regions of dyad symmetry including one just upstream of the promoter. The upstream regions of F and S genes were compared with those of five other androgen-responsive rodent genes in an attempt to identify common sequence motifs that might be involved in hormonal regulation of gene expression. PMID- 2987805 TI - Nucleotide sequence and biochemical characterization of the metJ gene from Salmonella typhimurium LT2. AB - The nucleotide sequence of the Salmonella typhimurium metJ gene is presented along with the sequence of the promoter region for the closely linked metB gene. The two genes are transcribed in opposite directions, with transcription initiating from a single promoter for metB, and from two apparent promoters for metJ. RNA polymerase binding sites for metJ and metB, determined by in vitro protection studies, lie adjacent to each other and may overlap. The two metJ promoters, PJ1 and PJ2, are separated by approximately 65 base pairs. Binding of RNA polymerase in vitro could only be observed for PJ1, even though transcripts are initiated from both promoters in vivo. The metJ gene codes for a polypeptide of 105 amino acids with a calculated Mr of 12,110. The translation start site was determined by N-terminal amino acid sequence analysis of a metJ-lacZ fusion protein. PMID- 2987806 TI - Measurement of gene number for seed storage proteins in Pisum. AB - We have measured the numbers of genes coding for the three seed storage proteins, vicilin, convicilin and legumin, in a number of Pisum genotypes of variant protein composition. No difference in gene number existed among P. sativum genotypes for any of the proteins. There were differences in the number of genes coding for individual proteins with approximately 11 genes (per haploid genome) for vicilin, 8 genes for legumin and 1 gene for convicilin. PMID- 2987807 TI - Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize. AB - The Adh2 gene of maize has a nucleotide sequence closely related to that of the maize Adh1 gene indicating that the two genes arose from a progenitor gene by a duplication event. The coding regions are 82% conserved at the nucleotide level and 87% conserved at the amino acid level. Each gene has nine introns in identical positions but their nucleotide sequences and lengths differ. Adh2 encodes a single mRNA and has a possible polyadenylation signal, AATAAT, fifteen bases upstream from the polyadenylation site. The Adh2 gene together with the Adh1 gene is induced by anaerobic conditions and under these conditions produces an increased level of mRNA. The 5' untranslated regions of the transcripts of Adh1 (100 bp) and Adh2 (126 bp) have diverged in sequence except for a conserved region which may be important for their anaerobic-specific translation. The 3' and 5' flanking sequences of the two genes are also divergent except for 11 bp of precise homology around the TATA boxes and three other 8 bp sequences further upstream. One of the common 8 bp sequences (CACCTCCC) is in a similar position ( 150 bp to -200 bp) in the two genes and may have a regulatory function in gene expression. PMID- 2987808 TI - Association of a truncated cytochrome c processed pseudogene with a similarly truncated member from a long interspersed repeat family of rat. AB - The cytochrome c multigene family of rat contains approximately 30 processed pseudogenes that represent genomic DNA copies of three alternate mRNAs. Here, the DNA sequence of an unusual processed pseudogene reveals that it has a complete 3' noncoding region including a short poly A tail but unlike the others is abruptly truncated at its 5' end, 19 amino acid codons from the translation terminator. At this position the pseudogene is fused through 17 consecutive adenylic acid residues to a 1.3 kb repetitive sequence. This repetitive element is flanked by direct repeats and represents a truncated member from a major long interspersed repeat family. The rat element is a composite of sequences observed in long interspersed repeats from both rodents and primates. Comparison to the equivalent mouse sequences shows that the 5' half of the repeat distal to the pseudogene has an open reading frame and is highly conserved whereas the half adjacent to the pseudogene is evolutionarily unstable. The proportion of cytochrome c pseudogene recombinant clones containing this repetitive DNA is 3 fold greater than observed in random isolates and may reflect a general tendency of processed pseudogenes to associate with other repetitive sequences in the genome. PMID- 2987809 TI - Beta-thalassemia resulting from a single nucleotide substitution in an acceptor splice site. AB - Beta-globin gene mutations which alter normal globin RNA splicing have confirmed the necessity of invariant nucleotides GT at donor splice sites. Functional consequences of point mutations in the invariant AG acceptor splice site have not been determined. We have isolated and characterized a beta-globin gene from a Black patient with beta-thalassemia intermedia which has an A-G transition at the usual intervening sequence 2 (IVS2) acceptor splice site. Functional analysis of transcripts produced by this mutant gene in a transient expression vector indicates that the mutation inactivates the normal acceptor splice site and results in some utilization of a cryptic splice site near position 580 of IVS2. This mutation would be expected to produce a beta-globin gene which results in no normal beta-globin mRNA. PMID- 2987811 TI - Characterization of a mouse interferon gene locus I. Isolation of a cluster of four alpha interferon genes. AB - A BALB/c mouse genomic library was screened with a murine interferon alpha 2 (MuIFN-alpha 2) cDNA coding region fragment. Eight clones were isolated which contain different mouse chromosomal segments related to the MuIFN-alpha 2 probe and a 28 kilobase (kb) region of mouse genomic DNA containing four different MuIFN-alpha genes (alpha 1, alpha 4, alpha 5 and alpha 6) was identified and characterized; an intergenic 1000 nucleotide long conserved sequence was found to be associated with three of these four alpha genes, indicating that this alpha IFN gene cluster evolved through tandem duplications. Sequence analysis revealed the absence of a polyadenylation site in the 3' untranslated region of MuIFN alpha 1, and showed that one of the genes (alpha 4) contains an internal deletion of 5 amino acids in the coding region. PMID- 2987810 TI - Organization, structure and expression of murine interferon alpha genes. AB - Using a human interferon-alpha probe we have isolated recombinant phages containing murine interferon-alpha (Mu IFN-alpha) genes from a genomic library. One of these phages contained two complete Mu IFN-alpha genes and part of a third gene. The insert of a second phage held two IFN genes. This indicates that the Mu IFN-alpha genes are clustered in the genome as is the case for the analogous human genes. The nucleotide sequences of these 5 genes were determined. They show that the genes are all different, albeit highly homologous. The deduced amino acid sequences show that four of the five genes contain a putative glycosylation site. Three genes were transiently expressed in COS cells and they gave rise to protein products showing antiviral properties. The expression of the five Mu IFN alpha genes and the Mu IFN-beta gene was studied in virus-induced mouse L cells. The individual mRNAs were visualized in a nuclease S1 experiment, using a specific probe for each gene. In RNA preparations from induced cells mRNAs for each of the five alpha genes and the beta gene were present. However, substantial differences in the amounts of the individual mRNAs were observed. PMID- 2987812 TI - Characterization of a mouse interferon gene locus II. Differential expression of alpha-interferon genes. AB - A cluster of four MuIFN-alpha genes was recently isolated and characterized (1); one of the genes in this cluster had, in the coding region, an internal deletion of 5 amino acids. Bacterial expression plasmids were constructed to examine the effect of this deletion on the antiviral activity of the MuIFN-alpha 4 peptide and it was found that the alpha 4 interferon peptide had a 100-fold lower antiviral activity than full length alpha-interferon proteins when expressed in E. coli. Three of the four MuIFN-alpha genes identified were expressed coordinately in L-cells infected with NDV. The relative levels of alpha 4 mRNA were substantially higher than the levels of the other alpha mRNAs. Comparison of the 5' end flanking sequences of these four alpha interferon genes revealed that the promoter sequences of alpha 1, alpha 5 and alpha 6 are more homologous to each other than to the alpha 4 promoter which also contains a G rich cluster not seen in the other three promoters. PMID- 2987813 TI - An ATP-dependent supercoiling topoisomerase of Chlamydomonas reinhardtii affects accumulation of specific chloroplast transcripts. AB - We have found that Chlamydomonas reinhardtii cells contain an ATP-dependent topoisomerase activity that supercoils circular DNA in vitro. Subsequent addition of a type I topoisomerase eliminates the supercoils. Like bacterial gyrase, this activity is inhibited by low concentrations of novobiocin (less than 0.1 microM) and by nalidixic acid (less than 0.1 microM). We have examined the effects of these topoisomerase inhibitors on accumulation of various chloroplast transcripts in vivo. Novobiocin differentially affected such transcripts; some transcripts became more abundant while many others were reduced in the presence of this drug. Nalidixic acid on the other hand caused many transcripts to become more abundant albeit to varying degrees. Inhibitors of this algal topoisomerase specifically stimulate a family of related transcripts which we have previously shown to be under light-dark control. We discuss how the inhibitors of this topoisomerase might exert their in vivo effects. PMID- 2987814 TI - Detailed analysis of the mRNAs mapping in the short unique region of herpes simplex virus type 1. AB - We have analysed the mRNAs which map within the short unique (US) region of the herpes simplex virus type 1 (HSV-1) genome. US has a total length of 12979 base pairs (1) and is extensively transcribed with approximately 94% of the total sequence present in cytoplasmic mRNAs and 79% of the total sequence considered to be protein coding. There are several examples of overlapping functions and multiple use of DNA sequence within this region. US contains 12 genes (1) which are expressed as 13 mRNAs. Two of these mRNAs are thought to arise from the same gene since they differ only slightly in the positions of their 5' ends and probably specify the same polypeptide. 11 of the 13 mRNAs are arranged into four nested families with unique 5' ends and common 3' co-termini. The other two mRNAs have unique 5' and 3' ends. PMID- 2987815 TI - Nucleotide sequence of a cluster of early and late genes in a conserved segment of the vaccinia virus genome. AB - The nucleotide sequence of a 7.6 kb vaccinia DNA segment from a genomic region conserved among different orthopox virus has been determined. This segment contains a tight cluster of 12 partly overlapping open reading frames most of which can be correlated with previously identified early and late proteins and mRNAs. Regulatory signals used by vaccinia virus have been studied. Presumptive promoter regions are rich in A, T and carry the consensus sequences TATA and AATAA spaced at 20-24 base pairs. Tandem repeats of a CTATTC consensus sequence are proposed to be involved in the termination of early transcription. PMID- 2987816 TI - Drosophila topoisomerase II double-strand DNA cleavage: analysis of DNA sequence homology at the cleavage site. AB - In order to study the sequence specificity of double-strand DNA cleavage by Drosophila topoisomerase II, we have mapped and sequenced 16 strong and 47 weak cleavage sites in the recombinant plasmid p pi 25.1. Analysis of the nucleotide and dinucleotide frequencies in the region near the site of phosphodiester bond breakage revealed a nonrandom distribution. The nucleotide frequencies observed would occur by chance with a probability less than 0.05. The consensus sequence we derived is 5'GT.A/TAY decrease ATT.AT..G 3', where a dot means no preferred nucleotide, Y is for pyrimidine, and the arrow shows the point of bond cleavage. On average, strong sites match the consensus better than weak sites. PMID- 2987817 TI - DNAaseI-hypersensitive minichromosomes of SV40 possess an elastic torsional strain in DNA. AB - Previously, we have shown that DNA in a small fraction (2-5%) of SV40 minichromosomes was torsionally strained and could be relaxed by treating minichromosomes with topoisomerase I. This fraction was enriched with endogeneous RNA polymerase II (Luchnik et al., 1982, EMBO J., 1, 1353). Here we show that one and the same fraction of SV40 minichromosomes is hypersensitive to DNAase I and is relaxable by topoisomerase I. Moreover, this fraction completely loses its hypersensitivity to DNAase I upon relaxation. The possibility that this fraction of minichromosomes can be represented by naked DNA is ruled out by the results of studying the kinetics of minichromosome digestion by DNAase I in comparison to digestion of pure SV40 DNA and by measuring the buoyant density of SV40 chromatin in equilibrium CsCl gradient. Our data obtained with SV40 minichromosomes may be relevant to the mechanism responsible for DNAase I hypersensitivity in the loops or domains of cellular chromatin. PMID- 2987818 TI - Escherichia coli 23S ribosomal RNA truncated at its 5' terminus. AB - In a strain of E. coli deficient in RNase III (ABL1), 23S rRNA has been shown to be present in incompletely processed form with extra nucleotides at both the 5' and 3' ends (King et al., 1984, Proc. Natl. Acad. Sci. U.S. 81, 185-188). RNA molecules with four different termini at the 5' end are observed in vivo, and are all found in polysomes. The shortest of these ("C3") is four nucleotides shorter than the accepted mature terminus. In growing cells of both wild-type and mutant strains up to 10% of the 23S rRNA chains contain the 5' C3 terminus. In stationary phase cells, the proportion of C3 termini remains the same in the wild type cells; but C3 becomes the dominant terminus in the mutant. Species C3 is also one of the 5' termini of 23S rRNA generated in vitro from larger precursors by the action of purified RNase III. We therefore suggest that some form of RNase III may still exist in the mutant; and since no cleavage is detectable at any other RNase III-specific site, the remaining enzyme would have a particular affinity for the C3 cleavage site, especially in stationary phase cells. We raise the question whether the C3 terminus has a special role in cellular metabolism. PMID- 2987819 TI - Characterization of a 3'----5' exonuclease activity in the phage phi 29-encoded DNA polymerase. AB - Purified protein p2 of phage phi 29, characterized as a specific DNA polymerase involved in the initiation and elongation of phi 29 DNA replication, contains a 3'----5' exonuclease active on single-stranded DNA, but not on double-stranded DNA. No 5'----3' exonuclease activity was found. The 3'----5' exonuclease activity was shown to be associated with the DNA polymerase since 1) the two activities were heat-inactivated with identical kinetics and 2) both activities, present in purified protein p2, cosedimented in a glycerol gradient. PMID- 2987820 TI - rDNA in Locusta migratoria is very variable: two introns and extensive restriction site polymorphisms in the spacer. AB - Cloned ribosomal DNA (rDNA) of Locusta migratoria was analyzed by restriction site mapping and SI nuclease experiments. The repeat unit is 18 kb long. The nontranscribed spacer region (NTS) is very large (11 kb) and homogeneous in length, but many of the restriction sites are heterogeneous among the repeat units. Two introns of different length were found at different positions in the 28S gene. They are present in less than 5% of the genes. PMID- 2987821 TI - Construction of a vector, pRSVcatamb38, for the rapid and sensitive assay of amber suppression in human and other mammalian cells. AB - We describe the generation of an amber mutation in the chloramphenicol acetyltransferase (cat) gene of the mammalian cell transfection vector pRSVcat (Gorman et.al. (1982), Proc.Natl.Acad.Sci. 79 6777-6791). We have demonstrated the in vivo suppression of this amber mutation in monkey and human cells by co transfection with a synthetic Xenopus suppressor tRNATyr under the control of the late SV40 promoter. The vector, pRSVcatamb38, may be used to quantitate amber suppression in various mammalian cells. PMID- 2987824 TI - Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. AB - We have isolated and sequenced a full-length cDNA clone encoding rat glyceraldehyde-3-phosphate-dehydrogenase (GAPDH, E.C.1.2.1.12). The entire mRNA is 1269 nucleotides long exclusive of poly(A) and contains respectively 71 and 196 bases of 5' and 3' non-coding regions. Primer extension as well as S1 nuclease protection experiments clearly established that a single (or at least a highly prominent) GAPDH mRNA species is expressed in all rat tissues examined. This sequence allowed the determination of the hitherto unknown primary structure of rat GAPDH which is 333 aminoacids long. Comparison between GAPDH sequences from rat, man and chicken revealed a high degree of sequence conservation at both nucleotide and protein levels. PMID- 2987825 TI - Stimulation of the UvrABC enzyme-catalyzed repair reactions by the UvrD protein (DNA helicase II). AB - An in vitro assay system was constructed using highly purified preparations of UvrA, UvrB, UvrC, UvrD proteins and DNA polymerase I, the objective being to analyse the role of UvrD protein in excision repair of UV-induced DNA damage. UvrABC enzyme-initiated repair synthesis was greatly enhanced by the addition of UvrD protein to the reaction mixture. Further analysis revealed that UvrD protein stimulated introduction of strand breaks in irradiated DNA by UvrABC enzyme but had no effect on the DNA polymerase I reaction. Thus, the site of action of UvrD protein is probably at the incision-excision step and not in later steps in excision repair. PMID- 2987822 TI - The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini. AB - Our previous DNA sequence comparisons of 3' terminal portions from equivalent herpes simplex virus type 1 (HSV-1) and HSV-2 genes identified a conserved sequence (consensus YGTGTTYY; Y = pyrimidine) located approximately 30bp downstream from the AATAAA signal. We report here that this signal is located downstream from 67% of the mammalian mRNA 3' termini examined. Using constructions with the bacterial chloramphenicol acetyl transferase (CAT) gene linked to an HSV 'terminator' fragment, we show that deletions in the 'terminator' reduce CAT activities and the levels of CAT mRNA 3' termini. Specifically: (1) deletions of downstream sequences which extend up to the consensus YGTGTTYY signal reduce CAT levels to values 35% of those obtained with undeleted plasmids, (2) a deletion of a further 14bp, which removes the YGTGTTYY consensus but not the poly A site, reduces CAT activities to 1%-4%. The levels of CAT mRNA 3' termini reflect the reductions in CAT activities however, levels of mRNA 5' termini are unaffected by these deletions. The RNA produced in the absence of the YGTGTTYY signal is present in the cytoplasm although no CAT activity is detectable. PMID- 2987823 TI - Differential activation of RNA polymerase III-transcribed genes by the polyomavirus enhancer and the adenovirus E1A gene products. AB - We have compared the effect of the polyomavirus cis-acting transcriptional enhancer and the adenovirus trans-acting E1A gene on expression of RNA polymerase III-transcribed genes (the adenovirus VAI gene and a bacterial tRNA gene) using DNA transfection and transient expression assays. The polyomavirus enhancer has little effect upon transcription of the VAI gene by RNA polymerase III in any cell type tested (murine, hamster, or human). In contrast, expression of the E1A gene within adenovirus infected cells stimulates transcription of RNA polymerase III-transcribed genes from co-transfected DNAs. Human 293 cells, which constitutively produce adenovirus E1A gene products, also express high levels of RNA polymerase III transcripts from transfected DNAs. PMID- 2987827 TI - Two new restriction endonucleases DraII and DraIII from Deinococcus radiophilus. AB - In addition to recently characterized DraI (1), two new Type II restriction endonucleases, DraII and DraIII, with novel site-specificities were isolated and purified from Deinococcus radiophilus ATCC 27603. DraII and DraIII recognize the hepta- and nonanucleotide sequences (sequence in text) The cleavage sites within both strands are indicated by arrows. The recognition sequences were established by mapping of the cleavage sites on pBR322 (DraII) and fd109 RF DNA (DraIII). The sequence specifities were confirmed by computer-assisted restriction analyses of the generated fragment patterns of the sequenced DNA's of the bacteriophages lambda, phi X174 RF, M13mp8 RF and fd109 RF, the viruses Adeno2 and SV40, and the plasmids pBR322 and pBR328. The cleavage positions within the recognition sequences were determined by sequencing experiments. PMID- 2987826 TI - Nucleotide sequence of mouse IL-2 receptor cDNA and its comparison with the human IL-2 receptor sequence. AB - We have cloned cDNA encoding the mouse interleukin-2 (IL-2) receptor from a murine T cell line, CTLL using human IL-2 receptor cDNA as probe. COS 7 cells transfected with the cDNA expressed the antigen recognized by the monoclonal antibody against the murine IL-2 receptor. The cDNA identified 4 species of mRNA (4.5, 3.5, 2.2 and 1.5 kb) of the mouse IL-2 receptor in CTLL cells. Difference in the length of mRNA seems to be ascribed to the variable length of the 3' untranslated sequence. Total nucleotide sequence (approximately 1400 bp) of this cDNA was determined and compared with the human receptor. The nucleotide and amino acid sequences of the IL-2 receptor are 70% and 60%, respectively, homologous in average between the two species. The comparison has revealed several conserved regions localized to particular exons such as transmembrane and cytoplasmic portions, suggesting that these regions are important for receptor function and its regulation. PMID- 2987828 TI - Topoisomerase I has a strong binding preference for a conserved hexadecameric sequence in the promoter region of the rRNA gene from Tetrahymena pyriformis. AB - Topoisomerase I is in situ associated with DNaseI hypersensitive sites located in the promotor and terminator regions of the extrachromosomal rDNA in Tetrahymena thermophila at sites with sequences fitting the motif (sequence in text) Reconstitution experiments with purified topoisomerase I and cloned fragments of rDNA demonstrate that the enzyme exhibits the same binding and cleavage properties on naked DNA. These observations are striking as topoisomerase I previously has been found to exhibit low sequence specificity. The specific binding of the enzyme has an absolute requirement for divalent cations with a preference for Ca2+. The strong binding to the hexadecamer has been characterized by competition experiments, and it has been used to determine the molecular weight of the enzyme. PMID- 2987829 TI - Respiratory syncytial virus fusion glycoprotein: nucleotide sequence of mRNA, identification of cleavage activation site and amino acid sequence of N-terminus of F1 subunit. AB - The amino acid sequence of respiratory syncytial virus fusion protein (Fo) was deduced from the sequence of a partial cDNA clone of mRNA and from the 5' mRNA sequence obtained by primer extension and dideoxysequencing. The encoded protein of 574 amino acids is extremely hydrophobic and has a molecular weight of 63371 daltons. The site of proteolytic cleavage within this protein was accurately mapped by determining a partial amino acid sequence of the N-terminus of the larger subunit (F1) purified by radioimmunoprecipitation using monoclonal antibodies. Alignment of the N-terminus of the F1 subunit within the deduced amino acid sequence of Fo permitted us to identify a sequence of lys-lys-arg-lys arg-arg at the C-terminus of the smaller N-terminal F2 subunit that appears to represent the cleavage/activation domain. Five potential sites of glycosylation, four within the F2 subunit, were also identified. Three extremely hydrophobic domains are present in the protein; a) the N-terminal signal sequence, b) the N terminus of the F1 subunit that is analogous to the N-terminus of the paramyxovirus F1 subunit and the HA2 subunit of influenza virus hemagglutinin, and c) the putative membrane anchorage domain near the C-terminus of F1. PMID- 2987830 TI - Long internal inverted repeat in a yeast viral double-stranded RNA. AB - The Saccharomyces cerevisiae viruses are non-infectious double-stranded (ds) RNA viruses present in most laboratory strains of yeast. Their genome consists of one or more dsRNAs separately encapsidated in particles composed mainly of one polypeptide, which has a Mr of 88 kdaltons in the best-studied viral subtype. A large viral dsRNA (L1, of 4.7 kb) encodes the capsid polypeptide. We have determined the sequences of a number of cDNA clones homologous to portions of L1 and mapped them by a novel heteroduplex technique. Several of these clones originate from a region of L1 2.3-2.5 kb from the 5' end of the plus strand that contains stop codons in all three reading frames in the plus strand. We therefore suspect that the capsid polypeptide gene lies in the 5' 2.3-2.6 kb of the plus strand. One of the cloned cDNAs has an inverted repeat of 170 bp that appears to be present in its parental RNA. The inverted repeat in L1 is the longest known inverted repeat in a viral dsRNA and the only known non-terminal inverted repeat. It might serve the function of creating two mRNAs from one viral dsRNA. PMID- 2987831 TI - Nucleotide sequence of a chicken delta-crystallin gene. AB - We have determined the complete nucleotide sequence of one of the two non-allelic delta-crystallin genes in the chicken, arbitrarily designated delta-gene 1, using a genomic clone (lambda g delta 106) containing the entire gene sequence. By comparison of the genomic sequence and the delta-crystallin cDNA sequence previously determined, we have identified exon sequences in the genomic sequence. Thus, the presence of 17 exons and 16 introns in the gene has been clarified. The delta-crystallin polypeptide deduced from the exon sequences consists of 465 amino acids which is larger, by 19 amino acid residues, than the polypeptide deduced from the cDNA sequence previously reported. Re-examination of the cDNA sequence using the same cDNA clone previously used shows that the present exon sequences are correct and the molecular weight of the deduced delta-crystallin polypeptide is 50,615 daltons instead of the previously reported value of 48,447 daltons. In addition, some structural features of the delta-crystallin gene including putative expression signals are discussed. PMID- 2987832 TI - Complete nucleotide sequence of a functional HLA-DP beta gene and the region between the DP beta 1 and DP alpha 1 genes: comparison of the 5' ends of HLA class II genes. AB - The complete nucleotide sequence of an HLA-DP beta 1 gene and part of the adjacent DP alpha 1 gene, up to and including the signal sequence exon, were determined. The sequence of the DP beta 1 gene identified it as the DPw4 allele. The six exons of the DP beta 1 gene spanned over 11,000 bp of sequence. The arrangement of the gene was broadly analogous to genes of other class II beta chains. The beta 1 exon was flanked by introns of over 4 kb. Comparisons with published sequences of cDNA clones indicated that an alternative splice junction, at the 3' end of the gene, is used in at least one allele. Variation in choice of splice junction indicates an additional mechanism for allelic variation in class II genes. The sequence also indicated that the DP beta 1 and DP alpha 1 genes are separated by only 2 kb at their 5' ends. Comparison of the 5' ends of the DP alpha 1 and beta 1 genes with other class II sequences, including the DZ alpha gene, showed conservation of several blocks of sequences thought to be involved in control of expression. Some areas of the introns were partially conserved in the DQ beta gene, and several other intron sequences were homologous to sequences found in other unrelated genes. PMID- 2987833 TI - Structure and expression of the ColE2-P9 immunity gene. AB - The primary structure and expression of the ColE2-P9 immunity gene (imm) were investigated. The imm gene is located behind the colicin gene (col) in the same orientation with an intergenic space of two base pairs. Although the imm gene was transcribed primarily in response to the SOS function of the host cell as well as the col gene, the immunity phenotype also appeared to be expressed by only a slight level of leaky transcription without an evident promoter. On comparing the ColE2-P9 sequence with those of relevant plasmids, a highly homologous sequence with the immE2 gene was found downstream of the immE3 gene of ColE3-CA38, and thus, an evolutional relationships could be deduced among some E-group Col plasmids. PMID- 2987835 TI - Temperature dependence of the gel electrophoretic mobility of superhelical DNA. AB - We have determined the gel electrophoretic behavior of closed circular plasmid pSM1 DNA (5420 bp) as a function of both temperature and of linking number (Lk). At temperatures below 37 degrees, the electrophoretic mobility first increases, then becomes constant as Lk is decreased below that of the relaxed closed DNA. As the temperature is increased above 37 degrees the electrophoretic mobility first increases as Lk decreases and then varies in a cyclic manner with further decreases in Lk. As the temperature is increased over the range 37 degrees - 65 degrees the cyclic behavior is manifested at progressively smaller decreases in Lk and the amplitude of the cycles increases. We interpret the results in terms of the early melting of superhelical DNA, in which the free energy associated with superhelix formation is progressively transferred to local denaturation. Using a two state approximation, we estimate the free energy change in the first cyclic transition to be 35 Kcal/mole DNA at 37 degrees and to decrease linearly with temperature. The free energy becomes equal to zero at a temperature of 71.6 degrees, which lies within 3 degrees of the melting temperature for the corresponding nicked circular DNA. From the slope of this relationship we estimate the apparent entropy and enthalpy of the first mobility transition to be 6.0 Kcal/mole base pair and 17.3 cal/mole base pair/degree, values consistent with duplex melting. PMID- 2987834 TI - Molecular structures of mitochondrial-DNA-like sequences in human nuclear DNA. AB - Two lambda phage clones carrying mitochondrial-DNA-like (mtDNA-like) sequences isolated from a human gene library were named Lm E-1 and Lm C-2, and their DNA structures were characterized. Lm E-1 contains about 0.4 kb DNA homologous to the 5' portion of the mitochondrial 16S ribosomal RNA (rRNA) gene and Lm C-2, a 1.6 kb DNA homologous to the 3' portion of the 12S rRNA gene and to almost all of the 16S rRNA gene. Comparisons of their nucleotide sequences with those of the corresponding regions of the human mtDNA revealed no detectable DNA rearrangement and their homologies to the human mtDNA are 84% and 80%, respectively. There are neither terminal repeats in the nuclear mtDNA-like sequences nor duplications of the nuclear DNAs flanking the mtDNA-like sequences. Evolutionary relationship between these two human nuclear mtDNA-like sequences and the human and bovine mtDNAs is discussed. PMID- 2987836 TI - Two divergent cellular src genes are expressed in Xenopus laevis. AB - Genomic and cDNA clones of the X. laevis src gene have been isolated and characterized by hybridization and DNA sequence analyses. The haploid genome of X. laevis contains two src genes, which can be distinguished from one another by virtue of sequence divergence in the 3' untranslated regions. Both of the genes are functional as indicated by the fact that oocytes contain RNAs transcribed from each of the genes. The two genes each encode an RNA which is 3.3 kb in length, or twice the length required to encode the 60,000 dalton src protein (pp60). Sequence analysis of the cDNA clones revealed that nearly all of the non coding sequence is located at the 3' end. The availability of sequence data from cDNA clones has also made it possible for the first time to identify with certainty the carboxyl terminal sequence of a cellular pp60 molecule. PMID- 2987838 TI - Analysis of regulatory sequences upstream of the E. coli uvrB gene; involvement of the DnaA protein. AB - A region located upstream of the uvrB promoters P1 and P2 was found to cause high plasmid loss when cloned in multicopy vectors. Two sequence elements responsible for this phenomenon were identified by mapping of spontaneous mutations that restore plasmid maintenance: a sequence known to have in vitro promoter activity and a partially overlapping sequence that shows extensive homology to recognition sites for the DnaA protein. Accordingly alterations in the level of DnaA protein in vivo were found to affect the extent of plasmid loss. A possible role for interaction of the DnaA protein with the region of interest is discussed in relation to regulation of uvrB expression. PMID- 2987837 TI - Structural organization of the P25 gene of Bombyx mori and comparative analysis of its 5' flanking DNA with that of the fibroin gene. AB - We have cloned a large portion of the P25 gene of Bombyx mori encoding the 25,000 dalton polypeptide which associates with fibroin to constitute the major silk protein. Its structure has been investigated by restriction mapping R-loop analysis, S1 nuclease protection experiments and nucleotide sequencing of the region spanning the 5' end of the gene and its flanking DNA. This has permitted a comparative sequence analysis of the DNA from the P25 and fibroin genes. The genes demonstrate no relatedness in their coding regions but they exhibit large blocks of sequence homology in their 5' flanking regions. In particular, the DNA upstream of the P25 gene possesses a sequence very similar to a region of fibroin 5' flanking DNA that is known to possess transcription modulation signals. The functional significance of these homologous regions is discussed with regard to the highly coordinated expression of these two genes. PMID- 2987839 TI - Nucleotide sequence of a functional cDNA for human thymidylate synthase. AB - We have determined the nucleotide sequence of a cDNA clone, pcHTS-1, encoding human thymidylate synthase (5,10-methylenetetrahydrofolate: dUMP C methyltransferase, EC 2.1.1.45) which was previously isolated from a human fibroblast expressible cDNA library and functional in mouse cells. The 1.6 kilobase cDNA insert of pcHTS-1 encodes a subunit protein of 313 amino acid (Mr = 35,706) and its predicted amino acid sequence is highly conserved in many regions including folylpolyglutamate and 5-fluoro-2'-deoxyuridylate binding sites, when compared with those of Lactobacillus casei, Escherichia coli, and bacteriophage T4. The cDNA contains in its 5'-untranslated region a triple tandemly repeated sequence consisting of 90 nucleotides, which starts immediately upstream of the ATG initiator codon, is very high in G+C content (80%), and can form three possible interconvertible stem-loop structures. PMID- 2987840 TI - The TGGCA protein binds to the MMTV-LTR, the adenovirus origin of replication, and the BK virus enhancer. AB - TGGCA-binding proteins are nuclear proteins with high affinity for double stranded DNA homologous to the prototype recognition sequence 5'YTGGCANNNTGCCAR 3'. Their ubiquitous tissue distribution in higher vertebrates characterizes them as a class of highly conserved proteins which may exert a basic function. To obtain clues to this function, specific binding sites were mapped on three viral genomes. Recognition sites were identified in the enhancer region of the BK virus, in the LTR of the mouse mammary tumor virus, and in the origin of replication of adenovirus 12. The TGGCA-binding protein from HeLa cells appears to be identical to nuclear factor I described by others, which stimulates initiation of adenovirus DNA replication in vitro. However, data from MMTV, BKV, and from cellular genes suggest that this specific protein-DNA interaction may also be involved in the control of gene activity. PMID- 2987842 TI - Effect of DNA damage on the expression of the chloramphenicol acetyltransferase gene after transfection into diploid human fibroblasts. AB - The activity of the chloramphenicol acetyltransferase (cat) gene after transfection into human fibroblasts has been measured following treatment of the plasmid pRSVcat with either restriction enzymes or ultraviolet light. Restriction enzymes producing single cuts in the plasmid inactivated the expression of the cat gene whether the enzymes cut the plasmid inside the coding region of the gene or several kilobases away from the gene. Ultraviolet light produced a dose dependent inactivation of the gene. The inactivation curve was steeper if the recipient cell strain was derived from a patient with xeroderma pigmentosum. The findings with this transient expression system contrast with previously reported results of experiments using plasmids which transform cells stably by integrating into the cellular genomic DNA. PMID- 2987841 TI - Cloning and nucleotide sequence of the aspartase gene of Escherichia coli W. AB - The aspA gene of Escherichia coli W which encodes aspartase was cloned into the plasmid vector pBR322. The nucleotide sequences of aspA and its flanking regions were determined. The aspA gene encodes a protein with a molecular weight of 52,224 consisted of 477 amino acid residues. The amino acid sequence of the protein predicted from the nucleotide sequence was consistent with those of the NH2- and COOH-terminal regions and also with the amino acid composition of the purified aspartase determined previously. Potential promoter and terminator sequences for aspA were also found in the determined sequence. PMID- 2987843 TI - Human rhinovirus 2: complete nucleotide sequence and proteolytic processing signals in the capsid protein region. AB - cDNA clones representing the entire genome of human rhinovirus 2 have been obtained and used to determine the complete nucleotide sequence. The genome consists of 7102 nucleotides and possesses a long open reading frame of 6450 nucleotides; this reading frame is initiated 611 nucleotides from the 5' end and stops 42 nucleotides from the polyA tract. The N-terminal sequences of three of the viral capsid proteins have been elucidated, thus defining the positions of three cleavage sites on the polyprotein. The extensive amino acid sequence homology with poliovirus and human rhinovirus 14 enabled the other cleavage sites to be predicted. Cleavages in the 3' half of the molecule appear to take place predominantly at Gln-Gly pairs, whereas those in the 5' half (including the capsid proteins) are more heterogeneous. PMID- 2987844 TI - Complete sequence of IS3. AB - We have determined the nucleotide sequence of IS3. Our IS3 isolate has 39 bp inverted repeats (IR's) with 6 mismatches, and is 1258 bp long. IS3 contains a large open reading frame (ORF) of 288 codons with a smaller, partially overlapping ORF of 91 codons on the opposite strand in codon-codon register. The large ORF is preceded by and has a 4 bp overlap with a 99 codon ORF that has potential transcriptional and translational start signals. Thus, IS3 could encode a bicistronic mRNA. The Shine-Dalgarno sequence for this 99 codon ORF could be sequestered in a stem-loop structure, but only if the transcript began outside IS3, as was first seen with IS10. This could be a means for preventing fortuitous activation of IS3 by outside promoters. No DNA sequence homology was found between IS3 and other prokaryotic IS elements, but there is slight amino acid sequence homology and significant conservation of hydropathicity patterns between the putative transposases of IS3 and IS2. PMID- 2987845 TI - Isolation and partial characterization of the entire human pro alpha 1(II) collagen gene. AB - Using a cDNA probe specific for the bovine Type II procollagen, a series of overlapping genomic clones containing 45 kb of contiguous human DNA have been isolated. Sequencing of a 54 bp exon, number 29, provided direct evidence that the recombinant clones bear human Type II collagen sequences. Localization of the 5' and 3' ends of the gene indicated that the human Type II collagen gene is 30 kb in size. This value is significantly higher than that of the homologous avian gene. The segregation of a polymorphic restriction site in informative families conclusively demonstrated that the Type II gene is found in a single copy in the human haploid genome. Finally, sequencing of a triple helical domain exon has confirmed that a rearrangement leading to the fusion of two exons occurred in the pro alpha 1(I) gene, following the divergence of the fibrillar collagens. PMID- 2987846 TI - Cleavage within an RNase III site can control mRNA stability and protein synthesis in vivo. AB - We report that processing at a cloned bacteriophage T7 RNase III site results in strong stabilization of the mRNA relative to the full-length transcript. In contrast, processing by RNase III of the bacteriophage lambda int transcript leads to rapid degradation of the messenger. It is proposed that the mode of cleavage within the RNase III site determines mRNA stability. Single cleavage leaves part of the phage T7 RNase III site in a folded structure at the generated 3' end and stabilizes the upstream mRNA whereas double cleavage at the lambda int site removes the folded structure and accelerates degradation. In addition, the processed transcript is as active a messenger as the unprocessed one and can direct protein synthesis for longer times. This increased efficiency is accompanied by a proportional (3-4 fold) increase in protein levels. In contrast, processing at the lambda int site reduces Int synthesis. Thus, processing may either stabilize mRNA and stimulate gene expression or destabilize a messenger and prevent protein synthesis. The end result appears to be determined by the mode of cleavage within the RNase III site. PMID- 2987847 TI - Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. III. Nucleotide sequence of some 10,000 base pairs in the origin region. AB - Approximately 10,000 nucleotides were sequenced in the oriC region of the Bacillus subtilis chromosome. The first replicating DNA strands are hybridized with a SalI-EcoRI fragment (nucleotide #1206-2954) in one direction (left to right) and an EcoRI-PstI fragment (#2949-4233) in the other. Seven open reading frames (ORF) accompanied with Shine-Dalgarno (SD) sequences were identified. ORF638 and ORF821 were identified as gyrB and gyrA genes respectively based on genetic evidences and amino acid sequence data. Comparison of amino acid sequences revealed that ORF44, ORF446, ORF378 and ORF323 are homologous with rpmH, dnaA, dnaN and recF of Escherichia coli, respectively. Thus, the organization of the ORFs from ORF44 to ORF638 resembles the organization of genes in the rpmH-gyrB region of the E. coli chromosome. Two non-coding regions characteristic for oriC signals were found near the site of initiation of the first replicating DNA. They are composed of repeating sequences whose consensus sequence TTAT(C/A)CACA is identical to that of 4 repeating sequences in the oriC of E. coli. PMID- 2987848 TI - Structure and function of the region of the replication origin of the Bacillus subtilis chromosome. IV. Transcription of the oriC region and expression of DNA gyrase genes and other open reading frames. AB - We have determined nucleotide sequence of some 10,000 base pairs (bp) in the oriC region of the Bacillus subtilis chromosome. Initiation sites of transcription from this region were determined in vivo by the S1-mapping method. Five major initiation sites were found in the leader sequences of five open reading frames (ORF) deduced from the nucleotide sequence. The sixth site is located inside the ORF323("recF"). Putative promoters were found for each transcript. Function of these promoters was demonstrated in Escherichia coli by the Maxi-cell method using appropriate fragments cloned in pBR vectors. Based on these results, genes in 10,000 bp oriC region are divided into 4 transcriptional units. GyrB composes one unit with two other ORFs, while gyrA constitutes a single unit by itself. The promoters for ORF446("dnaA") and ORF378("dnaN") are located within the putative signal sequences for oriC. Transcription from these promoters is dependent on a dna-initiation gene, dnaB. PMID- 2987849 TI - The wheat mitochondrial gene for apocytochrome b: absence of a prokaryotic ribosome binding site. AB - The wheat mitochondrial gene for apocytochrome b (CYB) has been identified by its hybridization to a yeast CYB probe and its nucleotide sequence has been determined. The wheat CYB sequence predicts a cytochrome b apoprotein of 398 amino acids; it is almost identical to that of maize but has ten additional amino acids at the carboxy terminus. No introns are present in the wheat CYB gene, but an internal segment of the gene is repeated at another genomic location. Transcript analysis reveals a single wheat CYB mRNA of approximately 2.4 kb with a long untranslated leader. Sequences upstream of the CYB coding region are very similar in wheat and maize but the stretch proposed to be a ribosome binding site in maize is not conserved in wheat. The corresponding leader regions of the wheat mitochondrial mRNAs for cytochrome oxidase subunits I and II also lack complementarity to the 3'-end of the small subunit rRNA. We conclude that alternative signals are involved in the initiation of translation in plant mitochondria. PMID- 2987850 TI - Primary sequence and partial secondary structure of the 12S kinetoplast (mitochondrial) ribosomal RNA from Leishmania tarentolae: conservation of peptidyl-transferase structural elements. AB - The sequence of the 1173 nt 12S kinetoplast ribosomal RNA from Leishmania tarentolae was determined from the maxicircle DNA sequence, and the 5' and 3' ends localized by primer runoff and S1 nuclease protection experiments. The gene was shown to be free of introns by S1 nuclease analysis. A partial secondary structure model of the 12S RNA molecule is presented which is equivalent in certain respects to the corresponding portions of the Escherichia coli 23S ribosomal RNA model. Domain II of the E. coli model is completely missing in the kinetoplast model with the exception of several phylogenetically conserved stems and one loop. There is a striking conservation of the functionally important peptidyl-transferase region except for the deletion of a few stems and loops. The 12S RNA is the smallest large subunit ribosomal RNA described to date. PMID- 2987851 TI - The nucleotide sequence of the RAD3 gene of Saccharomyces cerevisiae: a potential adenine nucleotide binding amino acid sequence and a nonessential acidic carboxyl terminal region. AB - The RAD3 gene of Saccharomyces cerevisiae is required for excision of pyrimidine dimers and is essential for viability. We present the nucleotide sequence of the RAD3 protein coding region and its flanking regions, and the deduced primary structure of the RAD3 protein. In addition, we have mapped the 5' end of RAD3 mRNA. The predicted RAD3 protein contains 778 amino acids with a calculated molecular weight of 89,779. A segment of the RAD3 protein shares homology with several adenine nucleotide binding proteins, suggesting that RAD3 protein may react with ATP. The twenty carboxyl terminal amino acids of RAD3 protein are predominantly acidic; however, deletion of this acidic region has no obvious effect on viability or DNA repair. PMID- 2987853 TI - A new method for sequence analysis of oligodeoxyribonucleotides. AB - The structure of an oligodeoxyribonucleotide may be determined by a simple two dimensional separation on a polyethyleneimine-cellulose thin layer sheet. Chromatography in the first dimension fractionates by chain length a nested set of fragments that are generated by subjecting the oligomer to partial spleen phosphodiesterase degradation and then labelling their non-common ends with 32P using polynucleotide kinase. A subsequent in situ treatment with nuclease Bal 31 produces labelled mononucleotides, and these are identified by chromatography in the second dimension. Since the method does not identify the 3' terminal nucleotide, a convenient procedure involving 3' end labelling followed by enzymatic digestion to monomers has been developed for this purpose. This approach to sequence analysis also has the advantage of permitting assignment of the identity and location of any modified or unusual bases within the oligonucleotide. PMID- 2987852 TI - The association of transcribed genes with the nuclear matrix of Drosophila cells during heat shock. AB - Using the transcriptional modulation afforded by heat shock, we found that the association of active genes with the nuclear matrix was not dependent on their level of transcription. Heat shock genes were matrix associated both before heat shock (when transcription was relatively low), and during heat shock (when transcription was greatly increased). Conversely, the cytoplasmic actin gene was matrix associated during normal growth conditions (when transcription was high) and during heat shock (when transcription was greatly decreased). Removal of greater than 99.7% of nascent RNA during preparation of the matrices did not affect these findings. Detailed examination of the cytoplasmic actin gene revealed that its matrix association was apparently mediated by multiple interactions near the 5' end of the gene. PMID- 2987854 TI - Deletion analysis of the Escherichia coli lactose promoter P2. AB - The Escherichia coli lactose (lac) operon transcription control region includes at least two sequences which are recognized by RNA polymerase holoenzyme in vitro, the normal lac promoter (termed P1) and an overlapping upstream promoter (termed P2). The structure of the P2 and the effect of RNA polymerase interaction at P2 on the association of RNA polymerase with P1 was analyzed by the isolation and characterization of various mutations at P2. A set of deletions with varying lengths of DNA between the lac P2 -10 region and a "-35 region" contributed by the vector DNA were constructed. In vitro studies indicate that as the spacing between the -10 region and "-35 region" is increased from 16 to 22 base pairs (bp), the steady state occupancy as measured by exonuclease III protection experiments and the ability to initiate transcripts from P2 decrease. Studies were also conducted using a single base pair insertion and a two base pair deletion between the natural -35 and -10 regions of P2. The mutation which decreases the in vitro occupancy and transcription initiation potential of P2 does not significantly affect the steady state in vitro occupancy of P1 nor the in vivo expression of the lac operon. These results are not consistent with the model that RNA polymerase occupancy at P2 competes with the P1 expression and therefore that this competition plays a role in cAMP bound catabolite gene activator protein (CAP-cAMP) control of the lac operon. PMID- 2987855 TI - Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. AB - Full length cDNAs encoding the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from rat and man have been isolated and sequenced. Many GAPDH gene-related sequences have been found in both genomes based on genomic blot hybridization analysis. Only one functional gene product is known. Results from genomic library screenings suggest that there are 300-400 copies of these sequences in the rat genome and approximately 100 in the human genome. Some of these related sequences have been shown to be processed pseudogenes. We have isolated several rat cDNA clones corresponding to these pseudogenes indicating that some pseudogenes are transcribed. Rat and human cDNAs are 89% homologous in the coding region, and 76% homologous in the first 100 base pairs of the 3' noncoding region. Comparison of these two cDNA sequences with those of the chicken, Drosophila and yeast genes allows the analysis of the evolution of the GAPDH genes in detail. PMID- 2987856 TI - The petunia chlorophyll a/b binding protein genes: a comparison of Cab genes from different gene families. AB - In Petunia (Mitchell) there are at least 16 genes which encode the chlorophyll a/b binding proteins; these genes have been classified into small multigene families based upon nucleotide sequence homology (1). A gene from each of five distinct Cab gene families is compared here. These genes have uninterrupted open reading frames of 266 or 267 amino acids corresponding to the Cab precursor proteins of sizes around 32000 daltons. A comparison of the amino acid sequences deduced here with published information from direct NH2-terminal analysis of a mature Cab protein in pea (10) suggests that a 34-36 amino acid transit peptide is cleaved from the NH2-terminal of the petunia precursor proteins. The proposed transit peptide sequences are more divergent than the mature peptide sequences between the Cab genes from different gene families. There are two regions within the mature Cab proteins which are conserved between all genes--a sequence of 28 amino acids near the NH2 terminal, and another sequence of 26 amino acids in the middle of the protein. The DNA sequences proximal to the Cab coding regions contain typical eukaryote promoter elements--TATA and CCAAT boxes, and in addition those genes which are known to be expressed in petunia leaf tissue also have an extensive region of homology (48 nucleotides) centered at approximately 130 nucleotides from the proposed transcription start sites. PMID- 2987857 TI - Analysis of the promoters for the two immunity genes present in the ColE3-CA38 plasmid using two new promoter probe vectors. AB - We have constructed two new promoter probe vectors which carry a polylinker derived from plasmid pUC19 proximal to the 5' end of a promoter-less galactokinase gene. Using these two vectors we have demonstrated that the ColE3imm gene and the ColE8imm gene present on the ColE3-CA38 plasmid have their own promoters, independent of the SOS promoter of the colicin E3 structural gene. The activity of two terminators, one located proximal to the 5' end of the ColE8imm gene, the other located proximal to the 5' end of the lys gene, were shown by a comparison of the galactokinase activity conferred by several of the recombinant plasmids. PMID- 2987858 TI - Nucleotide sequence of the 3' exon of the human N-myc gene. AB - We have analyzed a 3.8 kb Eco RI fragment of genomic DNA obtained from the amplified N-myc gene of human neuroblastoma cell line BE(2)-C. This fragment contains an exon with an open reading frame encoding approximately 170 amino acids of the carboxy-terminal end of the putative N-myc protein. Comparison of the inferred amino acid sequence of this peptide with that of the 3' domain of the human c-myc protein shows that locally conserved but dispersed regions of homology exist throughout the lengths of these peptides, while hydropathy plots indicate that the physical properties implied by their primary sequences are strikingly similar. Based upon these and other considerations, it is suggested that the 3' domains of c-myc and N-myc may potentially share related functions. PMID- 2987859 TI - alpha-Putrescinylthymine and the sensitivity of bacteriophage phi W-14 DNA to restriction endonucleases. AB - The modified base alpha-putrescinylthymine (putT) in phi W-14 DNA blocks cleavage of the DNA by 17 of 32 Type II restriction endonucleases. The enzymes cleaving the DNA do so to widely varying extents. The frequencies of cleavage of three altered forms of the DNA show that putT blocks recognition sites either when it occurs within the site or when it is in a sequence flanking the site. The blocking is dependent on both charge and steric factors. The charge effects can be greater than the steric effects for some of the enzymes tested. All the enzymes cleaving phi W-14 DNA release discrete fragments, showing that the distribution of putT is ordered. The cleavage frequencies for different enzymes suggest that the sequence CAputTG occurs frequently in the DNA. Only TaqI of the enzymes tested appeared not to be blocked by putT, but it was slowed down. TaqI generated fragments are joinable by T4 DNA ligase. PMID- 2987860 TI - Functional RNA polymerase II promoters in solitary retroviral long terminal repeats (LTR-IS elements). AB - LTR-IS elements are middle repetitive sequences in the mouse genome with structural features of solitary retroviral LTRs. In order to get some insight in the possible functional role of these sequences the promotor activity of two LTR IS representatives differing by 105 bp in their U3 region was investigated. Gene fusions between LTR-IS sequences and the bacterial gene coding for chloramphenicol acetyl transferase (CAT) were transfected into mouse 3T6 cells and the expression of CAT was measured. It is shown that the LTR-IS sequences represent weak RNA polymerase II promoters which require enhancement by cis-or trans-activating factors. PMID- 2987861 TI - Autonomously replicating sequences from the non transcribed spacers of Tetrahymena thermophila ribosomal DNA. AB - The non-transcribed spacer (NTS) regions of the linear extrachromosomal palindromic rDNA from the ciliated protozoan Tetrahymena thermophila contain, in addition to sequences regulating transcription, the origin(s) of bidirectional replication as well as telomere associated sequences. These NTS regions function as autonomous replication sequences (ARS) in the yeast Saccharomyces cerevisiae (Kiss, G.B., Amin, A.A. and Pearlman, R.E., Mol. Cell Biol. 1: 535-543, 1981). We have now identified in the 5' NTS two adjacent but non-overlapping restriction fragments which function as ARS in yeast. These fragments contain 200 bp of duplicated sequence and include the in vivo origin of rDNA replication. The ARS in the 3' NTS has been subdivided into a sequence which allows high frequency transformation of yeast but with transformants extremely unstable when grown either under selective or non-selective conditions, and a sequence which appears to be required for plasmid maintenance yet not to be essential for high frequency transformation. Detailed analysis of the DNA sequences in these regions gives rise to interesting information about structural and functional features of the molecule. PMID- 2987862 TI - Methyl phosphotriesters in alkylated DNA are repaired by the Ada regulatory protein of E. coli. AB - The E. coli ada+ gene product that controls the adaptive response to alkylating agents has been purified to apparent homogeneity using an overproducing expression vector system. This 39 kDa protein repairs 0(6)-methylguanine and 0(4) methylthymine residues in alkylated DNA by transfer of the methyl group from the base to a cysteine residue in the protein itself. The Ada protein also corrects one of the stereoisomers of methyl phosphotriesters in DNA by the same mechanism, while the other isomer is left unrepaired. Different cysteine residues in the Ada protein are used as acceptors in the repair of methyl groups derived from phosphotriesters and base residues. PMID- 2987863 TI - An empirical method for the evaluation of the quality of genomic DNA libraries. AB - An empirical method is presented that enables one to determine, with only a relatively small investment of time and materials, whether a lambda genomic DNA library will be a productive source of clones carrying specific sequences. The method provides an indication of the abundance of the sequences in library DNA and indicates whether major DNA rearrangements have occurred during library construction and amplification. PMID- 2987864 TI - In vitro transcription with extracts of nuclei of Drosophila embryos. AB - An in vitro transcription system has been developed from 0.3M NaCl extracts of nuclei of Drosophila embryos. Optimal transcription in the Drosophila embryo extract (DEX) was at 5mM MgCl2, 70mM KCl, 25 degrees C and with promoter concentrations of 0,75-1.0 pmol/assay. In vitro transcription from the Adenovirus 2 major late and the Drosophila histone gene promoters was studied in particular. S1-nuclease protection experiments showed that in vitro transcription from these promoters was accurate. In vitro transcription from the Adenovirus-2 major late promoter was less efficient than from histone gene H3 and H4 promoters in DEX. Vicecersa, in vitro transcription from Adenovirus-2 major late promoter was more efficient in HeLa whole cell extracts. The efficiencies of transcription from histone gene promoters decreased in DEX in the order H4 greater than or equal to H3 greater than H2a. Transcription from H2b and H1 promoters was not detected in DEX. The transcription from the Adenovirus-2 major late promoter was completely inhibited by histone H3 and H4 promoters. Preincubation of DEX with the adenoviral template, however, did not inhibit transcription from histone H3 and H4 promoters. The transcription start sites of histone genes H3 and H4 are separated by 160 base pairs. The H3 and H4 transcription start sites were subcloned separately. Now, a competition of transcription from the H3/H4 promoters with the Adenovirus-2 major late promoter was observed. The competition studies suggest that preincubation of DEX with the adenoviral template inhibited transcription from the H3 promoter more strongly than from the H4 promoter. PMID- 2987865 TI - Chromosome-specific alpha satellite DNA: nucleotide sequence analysis of the 2.0 kilobasepair repeat from the human X chromosome. AB - The pericentromeric region of the human X chromosome is characterized by a tandemly repeated family of 2.0 kilobasepair (kb) DNA fragments, initially revealed by cleavage of human DNA with the restriction enzyme BamHI. We report here the complete nucleotide sequence of a cloned member of the repeat family and establish that this X-linked DNA family consists entirely of alpha satellite DNA. Our data indicate that the 2.0 kb repeat consists of twelve alpha satellite monomers arranged in imperfect, direct repeats. Each of the alpha X monomers is approximately 171 basepairs (bp) in length and is 60-75% identical in sequence to previously described primate alpha satellite DNAs. The twelve alpha X monomers are 65-85% identical in sequence to each other and are organized as two adjacent, related blocks of five monomers, plus an additional two monomers also related to monomers within the pentamer blocks. Partial nucleotide sequence of a second, independent copy of the 2.0 kb BamHI fragment established that the 2.0 kb repeat is, in fact, the unit of amplification on the X. Comparison of the sequences of the twelve alpha X monomers allowed derivation of a 171 bp consensus sequence for alpha satellite DNA on the human X chromosome. These sequence data, combined with the results of filter hybridization experiments of total human DNA and X chromosome DNA, using subregions within the 2.0 kb repeat as probes, provide strong support for the hypothesis that individual human chromosomes are characterized by different alpha satellite families, defined both by restriction enzyme periodicity and by chromosome-specific primary sequence. PMID- 2987866 TI - Nucleotide sequence and characteristics of a Ty element from yeast. AB - We have determined the nucleotide sequence of a complete yeast Ty element (Ty pY109) which is located near a tRNA(Lys1) gene. The element is 5912 bp in length; the internal domain is flanked by two identical delta sequences of 331 bp. Ty pY109 contains two large open reading frames (ORFs) which overlap by 38 bp; the putative proteins consist of 440 and 1328 amino acid residues, respectively. The organisation of the coding sequences in Ty resembles that found in retroviral proviruses and the copia-like elements in Drosophila. Partial homologies have been found between Ty-ORF1 and tnpA from Tn3, and Ty-ORF2 and a reverse transcriptase-like domain (1,2). PMID- 2987867 TI - The human urokinase-plasminogen activator gene and its promoter. AB - The urokinase type of plasminogen activator (uPA) is subject to regulation by hormones, phorbol esters and oncogenic transformation. This enzyme has been suggested to play a key role in processes involving cell migration and tissue remodeling, and to be essential for tumor metastasis. In order to study these processes, we have isolated the human uPA gene, and have determined its entire nucleotide sequence. The gene is organized in 11 exons and is 6.4 kb long. The 5' end of uPA mRNA has been determined by both S1 mapping and primer extension experiments. A fragment of 800 bp containing the entire 5' flanking region shows promoter activity when introduced upstream of a bacterial chloramphenicol acetyltransferase gene and introduced into human cells. The hexanucleotide sequence GGCGGG, previously found at similar regions in several viral and eukaryotic promoters and shown to be essential for promoter activity (McKnight et al. (1984) Cell, 37, 253-262), is repeated three times between the CAAT and the TATA boxes. PMID- 2987868 TI - Cloning and nucleotide sequence of the tzs gene from Agrobacterium tumefaciens strain T37. AB - The trans-zeatin secretion locus (tzs), from the nopaline Ti plasmid of Agrobacterium tumefaciens strain T37, was cloned and the nucleotide sequence determined. This gene is located in the virulence region of pTiT37. The tzs gene is responsible for the secretion of trans-zeatin into bacterial culture medium and in addition has the cytokinin biosynthetic activity, dimethylallylpyrophosphate:AMP dimethylallyltransferase. Sequence analysis showed an open reading frame of 729 nucleotides, capable of encoding a protein of 27,545 daltons. A single new labelled protein of 27,200 daltons was detected in Escherichia coli maxicells expressing the cloned tzs gene. Significant sequence homology was observed between the tzs and the published tmr sequence from pTiT37. PMID- 2987869 TI - Preferential DNase I sensitivity of insert-free ribosomal RNA repeats of Drosophila melanogaster. AB - The five predominant types of rDNA repeats in D. melanogaster were analyzed with respect to their DNase I sensitivity. Only the insert-free repeats showed a generalized DNase I sensitivity pattern whereas the major type I, both minor type I and type II repeats were not as extensively degraded by the nuclease. For XX and XY embryonic nuclei, where there is rapid cell division, the majority of the In- repeats were DNase I sensitive. This indicated that these In- repeats have the potential to be transcribed during this developmental stage. When compared to the In- repeats, the chromatin configuration of the In+ repeats is indicative of a higher order of chromatin folding. The paucity of In+ primary gene transcripts observed in vivo could result from In+ repeats being packaged into a more condensed form of chromatin. PMID- 2987870 TI - Chemical synthesis of a gene for somatomedin C. AB - A synthetic gene for somatomedin C, a human growth factor, has been assembled by a single ligation of 23 oligodeoxyribonucleotides, which were chemically synthesized by an improved solid phase phosphotriester method. PMID- 2987871 TI - A mitochondrial reading frame which may code for a maturase-like protein in Saccharomyces cerevisiae. AB - In S. cerevisiae, the large oxi3/oli2 mitochondrial transcript contains the products of the oxi3, aap1 and oli2 genes and an unassigned reading frame, RF3. In the work presented here, we have completed the nucleotide sequence of RF3. We have shown that RF3 is composed of four fairly large ORFs which overlap within GC rich sequences. Furthermore, a shift of +1 base was found between each pair of consecutive reading frames. We discuss how these frameshifts could be removed to produce a 500 aminoacid long protein containing the two well conserved P1 and P2 oligopeptide sequences featuring several mitochondrial intron reading frames, suggesting, thereby, a RNA-maturase-like activity for the putative RF3 protein. In addition, we suggest that the insertion of GC clusters in a gene could provide a novel way of regulating its expression. PMID- 2987873 TI - Modification of the melting properties of duplex DNA by attachment of a GC-rich DNA sequence as determined by denaturing gradient gel electrophoresis. AB - The melting behavior of a DNA fragment carrying the mouse beta maj-globin promoter was investigated as a means of establishing procedures for separating DNA fragments differing by any single base substitution using the denaturing gradient gel electrophoresis procedure of Fischer and Lerman (1,2). We find that attachment of a 300 base pair GC-rich DNA sequence, termed a GC-clamp, to a 135 bp DNA fragment carrying the mouse beta-globin promoter significantly alters the pattern of DNA melting within the promoter. When the promoter is attached to the clamp, the promoter sequences melt without undergoing strand dissociation. The calculated distribution of melting domains within the promoter differs markedly according to the relative orientation of the clamp and promoter sequences. We find that the behavior of DNA fragments containing the promoter and clamp sequences on denaturing gradient polyacrylamide gels is in close agreement with the theoretical melting calculations. These studies provide the basis for critical evaluation of the parameters for DNA melting calculations, and they establish conditions for determining whether all single base substitutions within the promoter can be separated on denaturing gradient gels. PMID- 2987872 TI - Cloning and characterization of the DAS gene encoding the major methanol assimilatory enzyme from the methylotrophic yeast Hansenula polymorpha. AB - A gene library from the methanol utilizing yeast Hansenula polymorpha, constructed in a lambda Charon4A vector, was used to clone the gene encoding a key methanol assimilating enzyme, dihydroxyacetone synthase (DHAS) by differential plaque hybridization. The nucleotide sequence of the 2106 bp structural gene and the 5' and 3' non-coding regions was determined. The deduced amino acid sequence of the protein is in agreement with the apparent molecular weight and amino acid composition of the purified protein. The codon bias is not so pronounced as in some Saccharomyces cerevisiae genes. PMID- 2987874 TI - (TAA)n within sequences flanking several intrachromosomal variant surface glycoprotein genes in Trypanosoma brucei. AB - In Trypanosoma brucei telomeric copies of intrachromosomal variant surface glycoprotein (VSG) genes are produced through a gene conversion mechanism and are expressed if the telomere is or becomes transcriptionally activated. We have analyzed a sequence that occurs 1 to 1.5 kb 5' to several intrachromosomal VSG genes. This flanking sequence has three distinct sections: a 5' section containing 5 to 116 TAA or TAA-like tandem repeats; a moderately conserved sequence with an alternating GT characteristic; and a highly conserved 3' sequence that is largely alternating TA. Restriction mapping data and the location of this sequence suggest that it occurs at or near the 5' gene conversion boundary. This sequence, however, is not 5' to all VSG genes. These data suggest that this flanking sequence alone is not sufficient for VSG gene expression but that it may function in gene conversion for many, but not all, VSG genes. PMID- 2987875 TI - The plant ferredoxin precursor: nucleotide sequence of a full length cDNA clone. AB - A cDNA clone (pFD1) derived from Silene pratensis ferredoxin mRNA was selected from a cDNA-library using the hybrid released translation technique. Nucleotide sequence analysis showed the cDNA insert to contain the complete coding region of the ferredoxin precursor protein. The ferredoxin precursor has a mol.wt. of 15 300, the transit-peptide has a mol.wt. of 5600. The length of the ferredoxin mRNA was found to be 700 nucleotides whereas the cDNA insert was about 1200 basepairs. S1 nuclease protection experiments showed the ferredoxin-specific DNA to be 660 basepairs in length and to start 39 nucleotides upstream of the ferredoxin coding sequence. Southern blot analysis of genomic DNA revealed the presence of only one fragment with homology to the ferredoxin cDNA probe, so it is probably a single copy gene. Comparison of the ferredoxin transit-sequence with transit sequences of another stromal protein, the small subunit of ribulosebisphosphate carboxylase showed no apparent homology, except for a stretch of three amino acids near the processing site. PMID- 2987876 TI - Pea cytochrome oxidase subunit II gene has no intron and generates two mRNA transcripts with different 5'-termini. AB - We have isolated and sequenced the cytochrome oxidase subunit II (COII) gene from pea mitochondria. The coding sequence (777 bp) shows over 90% homology to the COII genes from three monocotyledonous plants (rice, maize and wheat) and one dicotyledonous plant (Oenothera berteriana). Several codons are deleted, however, in the pea COII gene. Of interest is the deletion in pea of the last three codons, including the stop codon, found at the 3' end of the other four COII genes. Instead, a new stop codon has been created due to a single-base substitution at the 13th bp downstream from the position of the original stop codon. This pea gene does not contain an intron which is found in all three monocots. Two distinct 5' termini of the pea COII transcripts have been identified by S1 nuclease mapping, one at 285 bp (site I) and the other at 302 bp (site II) upstream from the ATG codon. They are located at two identical sites within nearly perfect direct repeats. Transcripts with the 5' end corresponding to site I occur five time more frequently than those with the 5' end corresponding to site II. Both transcripts have the same 3' terminus which has been mapped to be at 193 to 195 bp downstream from the stop codon. PMID- 2987877 TI - In vivo transcription of rDNA spacers in Drosophila. AB - The sequence-nonhomologous rDNA spacers of D. melanogaster and D. virilis exhibit remarkably similar organizations and transcription capabilities. In spacers of both species, tandemly repeated sequences are present upstream of the beginning of the pre-rRNA coding region, and the repeats are capable of transcriptional activity. Short, nuclear transcripts homologous to the spacer repeats have been identified by S1 nuclease protection experiments and by northern blot analyses. Although spacer transcripts are rare in steady state RNA populations, the presence of multiple promoter elements may suggest a regulatory role for rDNA spacers. PMID- 2987878 TI - The divergent region of the Leishmania tarentolae kinetoplast maxicircle DNA contains a diverse set of repetitive sequences. AB - A 2.76 kb segment of the 12 kb divergent region of the Leishmania tarentolae kinetoplast maxicircle DNA consists almost entirely of repeated sequences. The repeats can be grouped into six families, some of which are present throughout the remainder of the divergent region. The repeats are oriented in a head-to-tail fashion with the three simplest repeats clustered into large arrays. A 47 bp palindrome and two copies of a "supercluster" of three different types of repeats are also present in the sequenced region. A sequence change in the divergent region is described for a clonal strain of L. tarentolae which was passaged continuously for several years. The repetitive sequences found in the divergent region appear to be appropriate substrates for the presumed deletion/insertion/recombination events occurring in this rapidly evolving portion of the maxicircle. PMID- 2987879 TI - Influence of monovalent cations on the activity of T4 DNA ligase in the presence of polyethylene glycol. AB - Monovalent cations such as Na+ and K+ inhibit the activity of T4 DNA ligase. However, the extent of inhibition varies with the terminal sequence of the duplex DNA used as substrate; in many cases, ligation of DNA is completely inhibited at 200 mM. The activity of the ligase is stimulated by raising the concentration of polyethylene glycol 6000 from 0 to 15% (w/v) when NaC1 and KC1 were both absent. Ligation was reduced as the concentration of NaC1 or KC1 was raised in a mixture containing 5 or 15% PEG 6000. With 10% PEG 6000, both cohesive- and blunt-end ligation of this ligase increased at high concentrations of salt (150-200 mM NaC1, or 200-250 mM KC1). Further, with 10% PEG 6000, inter- and intramolecular ligation occurred at low salt concentrations (0-100 mM NaC1, or 0-150 mM KC1); only linear oligomers were formed by intermolecular ligation at the high concentrations. PMID- 2987880 TI - Nucleotide sequences of three tRNA genes encoded in Tetrahymena mitochondrial DNA. AB - Nucleotide sequences of three cloned restriction fragments of Tetrahymena mtDNA which showed hybridization with mitochondrial tRNA have been determined. EcoRI fragment 5 (4.1 kbp) contains the tRNAphe gene sequence with anticodon GAA; Hind III fragment 6 (2.0 kbp) the tRNAhis with anticodon GTG; and EcoRI fragment 7 (1.9 kbp) the tRNAtrp with anticodon TCA. The CCA end is not encoded. All three tRNAs show usual features with common invariant and semi-invariant bases and can be folded into a cloverleaf structure with standard loops and regular base pairs in the stems. However, some minor irregular features are present including several GT pairs and an unmatched TT in the stems, and TCC instead of T psi C. All exhibit high G+C contents (about 50%); in contrast, the flanking regions are extremely A+T rich (about 80%). Several short coding frames can be deduced in these sequences, but their significance is not known. PMID- 2987881 TI - T4 DNA polymerase (3'-5') exonuclease, an enzyme for the detection and quantitation of stable DNA lesions: the ultraviolet light example. AB - Ultraviolet light irradiation of DNA results in the formation of two major types of photoproducts, cyclobutane dimers and 6-4' [pyrimidin-2'-one] -pyrimidine photoproducts. The enzyme T4 DNA polymerase possesses a 3' to 5' exonuclease activity and hydrolyzes both single and double stranded DNA in the absence of deoxynucleotide triphosphate substrates. Here we describe the use of T4 DNA polymerase associated exonuclease for the detection and quantitation of UV light induced damage on both single and double stranded DNA. Hydrolysis of UV irradiated single or double stranded DNA by the DNA polymerase associated exonuclease is quantitatively blocked by both cyclobutane dimers and (6-4) photoproducts. The enzyme terminates digestion of UV-irradiated DNA at the 3' pyrimidine of both cyclobutane dimers and (6-4) photoproducts. For a given photoproduct site, the induction of cyclobutane dimers was the same for both single and double stranded DNA. A similar relationship was also found for the induction of (6-4) photoproducts. These results suggest that the T4 DNA polymerase proofreading activity alone cannot remove these UV photoproducts present on DNA templates, but instead must function together with enzymes such as the T4 pyrimidine dimer-specific endonuclease in the repair of DNA photoproducts. The T4 DNA polymerase associated exonuclease should be useful for the analysis of a wide variety of bulky, stable DNA adducts. PMID- 2987882 TI - Conformational characteristics of the hexanucleoside pentaphosphate AUAUAU: a 2D NMR study at 500 MHz. AB - All 36 ribose proton resonances and most of the base proton resonances of the hexanucleoside pentaphosphate AUAUAU have been assigned unequivocally using 2D J resolved spectroscopy, spin echo correlated spectroscopy (SECSY) and 2D NOE spectroscopy (NOESY). The NMR parameters of AUAUAU are compared with those of smaller fragments that contain methylated adenine bases: m62AU, m62AUm62A, m62AUm62AU and m62AUm62AUm62A. Previous studies on this series of compounds have shown that in all these cases purine-pyrimidine-purine sequences prefer to adopt a mixture of states which have as common feature that the interior pyrimidine residues are bulged out, whereas the purine residues stack upon each other. Chemical shift data, proton-proton coupling constants, as well as the observation of imino-proton resonances for AUAUAU show unambiguously that upon lowering the temperature the high-temperature "bulged out" situation reverts to a normal A-RNA like double helix. PMID- 2987883 TI - Junction sequences generated by 'one-ended transposition'. AB - In the presence of the cognate transposase, plasmids containing a single inverted repeat (IR) sequence of Tn21 or of Tn1721 can fuse efficiently with other plasmids ('one-ended transposition'). The junctions across the sequences of donor and recipient DNA in recombinants generated by this process have been determined. These show that the segment of donor DNA starts precisely at the IR sequence (it is variable at the other end), and is flanked by a direct repeat of host DNA (usually 5bp) that was present only once in the original host sequence. These are characteristics of recombinants generated by transposition of Tn21 and Tn1721 themselves, suggesting that the mechanism of one-ended transposition is very similar to that of the corresponding entire elements. PMID- 2987884 TI - Effect of NusA protein on expression of the nusA,infB operon in E. coli. AB - Protein and operon fusions between lacZ and various genes of the nusA,infB operon have been constructed on lambda bacteriophages and used to show that the operon is negatively regulated by the level of NusA protein. Overproducing NusA (but not IF2) from a multicopy plasmid reduces the level of beta-galactosidase from the fusions indicating repression of the operon. Introducing the lambda carrying the fusions into nusA mutant strains produces a higher level of beta-galactosidase indicative of derepression of the operon. In particular, a larger form of the NusA protein which does not affect bacterial growth per se causes a derepression of the operon. As both protein and operon fusions respond equivalently, we conclude that the nusA protein is acting at the transcriptional level to regulate expression of the nusA, infB operon. PMID- 2987886 TI - The effect of site specific methylation on restriction endonuclease digestion. PMID- 2987887 TI - [Checklist for understanding magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS)]. AB - Overview on informations which are necessary to interpret MRI and MRS. The approach is related rather to application than it is a physical one. The central steps in comprehending MR are presented, e.g. frequency decomposition in order to obtain local information with MRI or chemical information with MRS, the spin echo in order to differentiate instrumental dephasation from dephasation by molecular interference, perfusion in MRI, and the components of sensitivity in measuring different nuclei etc. The report terminology is based on the description of grey scale pictures, but one has to consider that the information is not one dimensional but multiparametric. Therefore new terms have to be developed, a problem similar to that of describing scintigraphic time sequences in nuclear medicine. PMID- 2987888 TI - Clinical. Putting on the pressure. PMID- 2987889 TI - AIDS in children: a review of the clinical, epidemiologic and public health aspects. AB - A high level of suspicion by every provider is very important to ensure diagnosis and complete reporting. Such active surveillance will enable effective national monitoring of the occurrence of the disease, the definition of new risk groups and the identification of unusual cases for further study. Physicians and health care workers should report all children and adults suspected of having AIDS to their state health departments who, in turn, report to the CDC. It is clear that the number of cases of AIDS in children and adolescents is increasing because of either increased occurrence, increased diagnosis and/or increased surveillance. Pediatric health care personnel, especially those in high prevalence areas and those caring for high risk populations, need to be aware of AIDS and the problem encountered in caring for these children. PMID- 2987885 TI - Restriction and modification enzymes and their recognition sequences. PMID- 2987890 TI - [Post-transfusion cytomegalovirus hepatitis]. PMID- 2987891 TI - [Health status of workers chronically exposed to lead]. PMID- 2987892 TI - [Acquired immunodeficiency syndrome in children]. PMID- 2987895 TI - Maturation of the diurnal rhythm of corticosterone in female domestic fowl. AB - The purpose of this study was to examine the maturation of the avian corticosterone diurnal rhythm in relation to other indices of adrenocortical maturation. Female chickens were studied at 1, 5, 11, 17, 23, and 55 weeks of age. Blood samples were drawn at 4-hr intervals during a 24-hr (14 hr light: 10 hr dark) photoperiod. The diurnal rhythm in corticosterone concentration was not apparent until 11 weeks of age. At this age, unlike at 17 weeks of age, the maximal corticosterone concentration occurred during the light period. This indicates that although a rhythm is present at 11 weeks, a phase shift apparently occurs before the adult-like diurnal rhythm is attained. Injection of adrenocorticotropin (ACTH) produced an increase in the circulating corticosterone concentration in chickens at all ages. These results indicate 1) a phase shift must occur between the time of the initial emergence of a diurnal corticosterone rhythm and the attainment of the characteristic adult-like diurnal rhythm, and 2) the avian adrenal cortex is capable of responding to an ACTH stimulus with an increase in circulating corticosterone before the central nervous system rhythmically controls the pattern of circulating ACTH or corticosterone. PMID- 2987893 TI - Cytomegalovirus hemorrhagic colitis in a nontransplant patient. AB - A 75-year-old man with major colonic hemorrhage was found at exploratory laparotomy and colectomy to have acute sigmoid colitis induced by cytomegalovirus (CMV) infection. The usual risk factors for CMV colitis (previous mucosal injury, cytotoxic drug therapy, and transplantation) were absent. The stress and debility of an acute uremic state and a long hospital stay may have compromised the patient's immune system and allowed for viral invasion of the colon. This manifestation of cytomegalic disease is an extremely unusual but important clinical entity. Accurate and prompt diagnosis, which requires a high index of suspicion, is important, since in the majority of cases, colectomy is indicated for control of hemorrhage. PMID- 2987894 TI - Hirsutism. Pathophysiology, clinical evaluation, treatment. AB - Hirsutism is generally an androgen-mediated disorder. Tremendous progress has been made in elucidating the numerous clinical disorders that can cause it. Systematic evaluation of hirsute women must be directed at determining the cause of hyperandrogenemia, which in turn allows specific and effective therapy to be initiated. PMID- 2987896 TI - Vaccination against Marek's disease and infectious bursal disease III. Growth rate of both turkey herpesvirus and infectious bursal disease virus in coinfected cell cultures. AB - Titers of the turkey herpesvirus (HVT) and infectious bursal disease virus (IBDV) in coinfected cell cultures, used to produce a live bivalent HVT/IBDV vaccine, were determined daily. Based on the daily titers, cells harvested either 2 or 3 days after inoculating the second virus (IBDV) into HVT infected cell cultures yielded the maximum titers for both viruses. Harvesting coinfected cell cultures on either day provided the most suitable source of the live HVT/IBDV vaccine against Marek's disease and infectious bursal disease. PMID- 2987897 TI - [Pregnancy under converting enzyme inhibitor]. PMID- 2987898 TI - [Bowenoid papulosis of the cervix uteri]. PMID- 2987899 TI - [Quick detection of anticytomegalovirus antibodies]. PMID- 2987900 TI - [Neurotoxicity of vidarabine in chronic active hepatitis]. PMID- 2987901 TI - [Supratentorial glioblastoma in adults. Therapeutic results]. AB - This retrospective study concerns 55 adult patients with supratentorial glioblastoma. The tumours were treated by complete or partial surgical excision whenever possible (31 cases), radiotherapy (22 cases) in doses of 60 Gy over 6 to 7 weeks (40 Gy with telecobalt and 20 Gy with superimposed electrons) and multiple chemotherapy (10 cases) with VM26 and CCNU or BCNU. Although the number of patients in some categories was too small for statistical evaluation, the results obtained were in agreement with those found in the literature and indicative of what can be expected. In patients with inoperable tumours the mean survival was increased from 2 to 8 months by radiotherapy or chemotherapy given separately, and from 2 to 9 or 10 months only when these two methods were combined. The mean survival of patients with partial tumoral excision was 2.5 months extended to 10 months after post-operative radiotherapy; one patient in this group who received both radiotherapy and chemotherapy is still alive after more than 5 years. In patients with macroscopically satisfactory excision, the 12 months good quality survival obtained by surgery was apparently prolonged to 22 months with radiotherapy; 3 of these patients who had both radiotherapy and chemotherapy after surgery are alive and in good condition after 15, 16 and 28 months respectively. PMID- 2987902 TI - [Alternating chemotherapy and radiotherapy in the induction treatment of small cell bronchial carcinoma (forms limited to the thorax)]. AB - Sixty three patients with limited small cell lung carcinoma were entered into a pilot study alternating monthly cycles of combination chemotherapy (doxorubicin, VP16213, cyclophosphamide and methotrexate (group A) or cis platinum (group B) with 3 courses of mediastinal radiotherapy. The total mediastinal dose was 45 Gy for the first 28 patients (group A) and 55 Gy for the remaining 35 (group B). The complete response rate was 86% in group A (median survival 14 months) and 91% in group B (median survival 20 months). Local control at two years was 61% in group A and 82% in group B, while relapse-free 2 year survival rates were 32% and 37% respectively. The acceptable toxicity and high response rate of this combined modality therapy lead us to further research in maintenance therapy. PMID- 2987903 TI - GABA dysfunction in the pathophysiology of tardive dyskinesia. AB - Pharmacologic treatments which diminish central dopaminergic transmission improve symptoms of tardive dyskinesia (TD). These clinical data, supported by results from animal model studies, have provided a basis for the dopamine (DA) receptor hypersensitivity hypothesis of TD. Since its initial formulation, however, knowledge of the multiple effects of prolonged neuroleptic administration in mammalian CNS has greatly expanded. Clinical and animal model studies carried out independently now both suggest that GABA-mediated neuronal tracts of the basal ganglia are important, perhaps pivotal, in TD. Thus, we would extend the DA hypothesis of TD to include the idea that neuroleptic-induced DA receptor hypersensitivity in striatum results in GABA system hypofunction in striatal projection areas in those individuals who develop TD. PMID- 2987904 TI - Pathophysiology of L-dopa-induced abnormal involuntary movements. AB - Among the various deficiencies in neurotransmitters and neuropeptides in the brains of patients with Parkinson's disease, the loss of dopamine (DA) is implicated in a major way in the occurrence of L-dopa-induced abnormal involuntary movements (AIMs). Whatever the clinical pattern, they are triggered by drugs which stimulate DA transmission and can be modified by DA agonists and antagonists. They occur when DOPA plasma concentrations, and thus central DA receptor stimulation, reach a critical level. They are observed in patients with severely damaged central DA neurons, but involvement of other neurotransmitter containing cells cannot be excluded. L-Dopa-induced AIMs have clinical and somatotopic characteristics, which vary from patient to patient. One might speculate that variable damage to DA neurons, associated or not with other neurotransmitter-containing cells in the affected brain structures, causes these differences in AIM patterns. By analogy with behavioral experiments in animals, the hypersensitivity of DA receptors observed in the basal ganglia of parkinsonian patients post mortem might reasonably be considered to mediate L dopa-induced AIMs. However, the role of various subtypes of DA receptors or of changes in DA metabolism in the cell bodies and dendrites (substantia nigra) or nerve terminals (striatolimbic areas) must also be considered. In brief, the features, topography, and timing of L-dopa-induced AIMs are dependent upon alterations of the functional expression of striatal DA output, which is not yet well understood. PMID- 2987905 TI - Induction and reversal of dopamine dyskinesia in rat, cat, and monkey. AB - Abnormal involuntary movements (AIMs, stereotyped or dyskinetic movements) were induced with different dopamine mimetics in rat, cat, and monkey. In the rat only stereotyped movements were observed, whereas in the cat dopamine agonists (apomorphine) preferentially induced dyskinesia but dopamine/noradrenaline uptake inhibitors (d-amphetamine, nomifensine) induced predominantly stereotypes; L-dopa induced an equal, low, number of both kinds of movements in the cat. In the monkey with bilateral lesions of the nigrostriatal dopamine pathways the AIMs could be divided into type 1 dyskinesia (behavioral), type 2 dyskinesia (oral and psychomotor), and chorea. GABA agonists (progabide, muscimol) had a biphasic action on apomorphine stereotypes in the rat, slightly (10%-20%) augmenting these movements at low doses and antagonizing (greater than 50%) them at higher doses. As these latter doses of progabide also antagonize apomorphine-induced circling in rats with a unilateral lesion of the substantia nigra, it is likely that this action is exerted at or beyond the dopamine target cell. In cats the dyskinetic movements induced by apomorphine were abolished by progabide. In contrast, L-dopa induced stereotypies were resistant to the antidyskinetic action of progabide, and at low doses of L-dopa an increased incidence of stereotypies was noted. In the monkey, the type 1 dyskinesia following L-dopa and piribedil were also relatively resistant to progabide administration, whereas the type 2 dyskinesia and chorea were abolished by progabide. These studies are parallel to and support the clinical observations that dyskinetic movements following a direct action at the dopamine receptor (tardive dyskinesia) may be reversed by progabide whereas those associated with dopamine neuron activity, perhaps together with noradrenergic activation (L-dopa dyskinesia), are resistant to the antidyskinetic action of progabide. PMID- 2987906 TI - Influence of GABA mimetics and lithium on biochemical manifestations of striatal dopamine target cell hypersensitivity. AB - The potential mechanisms whereby GABA mimetics and the antimanic agent lithium stabilize dopaminergic transmission are discussed. Evidence is presented that GABA mimetics, and in particular progabide, affect dopamine-mediated events in the basal ganglia on at least three levels. First, they reduce dopamine neuron activity in both the basal and the activated states. Secondly, on a long-term basis, they antagonize the proliferation of striatal dopamine receptors subsequent to chronic neuroleptic treatment. Thirdly, they modulate the expression of dopamine receptor activation by acting distally to the dopaminergic synapse. Lithium and GABA mimetics have the last two properties in common. These effects may represent the biochemical basis for the therapeutic action of GABA mimetics in iatrogenic dyskinesias. Moreover, the similarity between the biochemical effects of GABA mimetics and lithium suggest that the former drugs may have a therapeutic potential in mania. PMID- 2987907 TI - New aspects on the role of dopamine, acetylcholine, and GABA in the development of tardive dyskinesia. AB - In this paper various new findings on the possible anatomical substrates of tardive dyskinesia will be presented. The results show that the striatum is heterogeneously organized, and the syndromes of biting, gnawing, and licking activities in the rat model involve a complex balance between various dopamine (DA), cholinergic, and GABAergic systems within the striatum and the mesolimbic and mesocortical systems. PMID- 2987908 TI - Tumours of minor oral salivary glands. PMID- 2987909 TI - Low-temperature magnetic circular dichroism studies of native laccase: spectroscopic evidence for exogenous ligand bridging at a trinuclear copper active site. AB - The detailed nature of N-3 binding at the multi-copper active site in native laccase is investigated through a combination of low-temperature magnetic circular dichroism (LTMCD) and absorption spectroscopies. This combination of techniques allows charge-transfer spectral features associated with N-3 binding to the paramagnetic type 2 Cu(II) to be differentiated from those associated with binding to the antiferromagnetically coupled, and therefore diamagnetic, binuclear type 3 Cu(II) site. Earlier absorption titration studies have indicated that N-3 binds with two different binding constants, yielding a high-affinity and a low-affinity form. The studies presented here are interpreted as strong evidence that low-affinity N-3 bridges the paramagnetic type 2 and diamagnetic type 3 binuclear Cu(II) sites in fully oxidized laccase. This assignment is further supported by features in the MCD spectrum whose intensity correlates with an EPR signal associated with uncoupled type 3 Cu(II) sites. In these sites, N-3 has displaced the endogenous bridge, thereby rendering the site paramagnetic and detectable by both LTMCD and EPR spectroscopy. High-affinity N-3 is found to bind to the paramagnetic type 2 Cu(II) site in a limited fraction of the protein molecules that contains reduced type 3 sites. Finally, the possible role of this trinuclear (type 2-type 3) Cu(II) active site in enabling the irreversible reduction of dioxygen to water is considered. PMID- 2987910 TI - 1H homonuclear editing of rat brain using semiselective pulses. AB - We have used a semiselective Hahn spin-echo sequence of the form (1331)-tau (2662)-tau-AQ, delivered by a surface coil to obtain high-resolution 1H NMR spectra from the brains of intact dead rats. This sequence gave suppression of the tissue water resonance by a factor of 80,000 when tau = 68 ms. Delivery of a frequency-selective Dante pulse train to the alpha-CH resonance of lactate at 4.11 ppm, simultaneously with the 2662 refocusing pulse, altered the j-modulation in the spin-coupled beta-CH3 protons. Subtraction of this spectrum from one in which the Dante was ineffective gave an edited spectrum containing only the beta CH3 resonance of lactate at 1.31 ppm. When the position of the Dante was shifted to 3.78 ppm to selectively invert the alpha-CH protons of alanine, an edited spectrum of alanine was obtained. PMID- 2987911 TI - A cosmid vector that facilitates restriction enzyme mapping. AB - We describe the construction and use of a cosmid vector, loric, which is derived from the phage lambda origin of replication and appears to be more stable than ColE1-derived cosmids. Loric recombinants can be efficiently packaged in vivo to yield 100-300 micrograms of DNA per liter that is linear and has single-stranded cos ends. We call such molecules "phosmids." Phosmid restriction maps can be rapidly generated by labeling either the left or right cos site by annealing on a 32P-labeled oligonucleotide complementary to either cos-L or cos-R. Partial restriction enzyme digestion, agarose gel electrophoresis, and autoradiography are used to size restriction fragments of increasing length, all of which terminate at the labeled cos site. The procedures have been tested by isolating and mapping a region of the H-2 locus of mouse chromosome 17. PMID- 2987912 TI - Phosphorylation of DNA topoisomerase II by casein kinase II: modulation of eukaryotic topoisomerase II activity in vitro. AB - The phosphorylation of Drosophila melanogaster DNA topoisomerase II by purified casein kinase II was characterized in vitro. Under the conditions used, the kinase incorporated a maximum of 2-3 molecules of phosphate per homodimer of topoisomerase II. No autophosphorylation of the topoisomerase was observed. The only amino acid residue modified by casein kinase II was serine. Apparent Km and Vmax values for the phosphorylation reaction were 0.4 microM topoisomerase II and 3.3 mumol of phosphate incorporated per min per mg of kinase, respectively. Phosphorylation stimulated the DNA relaxation activity of topoisomerase II by 3 fold over that of the dephosphorylated enzyme, and the effects of modification could be reversed by treatment with alkaline phosphatase. Therefore, this study demonstrates that post-translational enzymatic modifications can be used to modulate the interaction between topoisomerase II and DNA. PMID- 2987914 TI - Evidence for conduction of protons along the interface between water and a polar lipid monolayer. AB - Movements of H+ along the polar heads of phospholipids spread in monolayers were compared to movements of H+ in the aqueous subphase. The probe for detecting H+ movement along the monolayer was a pH-sensitive fluorescein chromophore covalently bound to the head group of phosphatidylethanolamine. The behavior of this probe was not affected by the electrical properties of the lipid/water interface. Lateral diffusion of H+ along the phospholipid/water interface was then studied by acid-jump experiments in which advantage was taken of the large size of the monolayer. H+ was injected a few centimeters away from the probe observation area. The time needed for H+ diffusion to the probe was monitored by the change in the fluorescence signal, fluorescein being nonfluorescent in an acid medium. Diffusion of H+ in the bulk phase was monitored by the fluorescence change of water-soluble fluorescein isothiocyanate. Diffusion along the lipid monolayer was found to be 20 times faster than in the bulk water phase and required a structured monolayer in order to occur, as revealed by variation of the molecular area occupied by the lipid molecules. The molecular basis of rapid H+ transfer along the lipid monolayer may be the existence of a hydrogen-bond network along the polar heads, capable of supporting a rapid "hop and turn" of H+. PMID- 2987913 TI - Beta-endorphin-(1-27) is a naturally occurring antagonist to etorphine-induced analgesia. AB - The potent opioid peptide beta-endorphin is found in the brain and pituitary with two related fragments, beta-endorphin-(1-27) and beta-endorphin-(1-26). The fragments retain substantial opioid-receptor binding activity but are virtually inactive analgesically. beta-Endorphin-(1-27) inhibits beta-endorphin-induced and etorphine-induced analgesia when coinjected intracerebroventricularly into mice. Antagonism by competition at the same site(s) is suggested from parallel shifts of the dose-response curves of etorphine or beta-endorphin in the presence of beta-endorphin-(1-27). Its potency is 4-5 times greater than that of the opiate antagonist naloxone. beta-Endorphin-(1-26) does not antagonize the antinociceptive action of etorphine or beta-endorphin in doses up to 500 pmol per animal. PMID- 2987915 TI - The rate-determining step in cAMP-mediated exocytosis in the rat parotid and submandibular glands appears to involve analogous 26-kDa integral membrane phosphoproteins. AB - The possible direct involvement of protein phosphorylation in the regulation of exocytosis during beta-adrenergic receptor stimulation in rat parotid and submandibular salivary glands was investigated in vitro using dispersed cells. The dispersed cells were labeled with [32P]orthophosphate for 40 min prior to experimental manipulation. Subcellular fractions were isolated, the proteins were separated using sodium dodecylsulfate/polyacrylamide gel electrophoresis (NaDodSO4/PAGE), and the phosphoproteins were detected by autoradiography. Changes in the extent of phosphorylation for each phosphoprotein were determined indirectly by densitometric analyses. The analogous parotid and submandibular 26 kDa membrane phosphoproteins had a rapid phosphate turnover rate (t 1/2 = 5-6 min) whereas the analogous 21-kDa membrane phosphoproteins had a much slower phosphate turnover rate (t 1/2 greater than 20 min). The results of Triton X-114 extraction indicated that the 26- and 21-kDa phosphoproteins were integral membrane proteins. The rate of phosphate turnover for the analogous 26-kDa phosphoproteins is compatible with a regulatory role in exocytosis, whereas the slower phosphate turnover rate for the analogous 21-kDa phosphoproteins suggests that these proteins may play a more subordinate role in secretion or they may coordinate secretion with other cellular metabolic events. PMID- 2987916 TI - Molecular cloning and nucleotide sequence of the alpha and beta subunits of allophycocyanin from the cyanelle genome of Cyanophora paradoxa. AB - The genes for the alpha- and beta-subunit apoproteins of allophycocyanin (AP) were isolated from the cyanelle genome of Cyanophora paradoxa and subjected to nucleotide sequence analysis. The AP beta-subunit apoprotein gene was localized to a 7.8-kilobase-pair Pst I restriction fragment from cyanelle DNA by hybridization with a tetradecameric oligonucleotide probe. Sequence analysis using that oligonucleotide and its complement as primers for the dideoxy chain termination sequencing method confirmed the presence of both AP alpha- and beta subunit genes on this restriction fragment. Additional oligonucleotide primers were synthesized as sequencing progressed and were used to determine rapidly the nucleotide sequence of a 1336-base-pair region of this cloned fragment. This strategy allowed the sequencing to be completed without a detailed restriction map and without extensive and time-consuming subcloning. The sequenced region contains two open reading frames whose deduced amino acid sequences are 81-85% homologous to cyanobacterial and red algal AP subunits whose amino acid sequences have been determined. The two open reading frames are in the same orientation and are separated by 39 base pairs. AP alpha is 5' to AP beta and both coding sequences are preceded by a polypurine, Shine-Dalgarno-type sequence. Sequences upstream from AP alpha closely resemble the Escherichia coli consensus promoter sequences and also show considerable homology to promoter sequences for several chloroplast-encoded psbA genes. A 56-base-pair palindromic sequence downstream from the AP beta gene could play a role in the termination of transcription or translation. The allophycocyanin apoprotein subunit genes are located on the large single-copy region of the cyanelle genome. PMID- 2987917 TI - Mitochondrial modulation of maternally transmitted antigen: analysis of cell hybrids. AB - Maternally transmitted antigen (Mta) is a murine cell surface class I-like antigen that is defined by specific cytotoxic lymphocyte reactivity. Mta is unique in that its expression requires cooperation between genetic elements both in the Qa/Tla region of chromosome 17 and in the cytoplasm. In view of the known cytoplasmic, and thus maternal, inheritance of mitochondria, we have directly assessed their potential involvement in Mta expression. The mitochondria-specific lethal dye rhodamine 6G (R6G) was used to control the input of mitochondria into cell hybrids. The parental lines, one of BALB/c and one of NZB origin, were known to differ in Mta and mtDNA phenotype. Our data show that most control BALB/c-NZB hybrids expressed the BALB/c Mta phenotype and likewise contained only BALB/c type mtDNA. The NZB Mta phenotype was not coexpressed in the control hybrids. However, when the mitochondrial contribution from BALB/c was prevented by R6G treatment, the majority of the resultant hybrids expressed only the NZB Mta type and likewise contained only NZB mtDNA. The exceptional R6G-treated hybrids that continued to express the BALB/c Mta phenotype likewise contained only BALB/c mtDNA. Thus, in every case the mtDNA phenotype correlated with the Mta phenotype of the cells. Together, the data support the remarkable conclusion that mitochondria modulate the phenotypic expression of a cell surface molecule. PMID- 2987918 TI - Friend murine leukemia virus-immortalized myeloid cells are converted into tumorigenic cell lines by Abelson leukemia virus. AB - Friend murine leukemia virus (Fr-MuLV) is a replication-competent murine retrovirus that induces acute nonlymphocytic leukemias in NFS/n mice. Fr-MuLV disease is divided into two stages based on the ability of the leukemia cells to grow in culture and transplant into syngeneic mice. Hematopoietic cells taken from the early stage of disease after Fr-MuLV infection grow as immortal myeloid cell lines in the presence of WEHI-3 cell-conditioned medium (CM) or interleukin 3. These growth factor-dependent cell lines do not grow in culture in the absence of CM and do not form tumors in syngeneic animals. If these Fr-MuLV-infected cells are superinfected with Abelson murine leukemia virus (Ab-MuLV), they lose their dependence on WEHI-3 CM and proliferate in culture in the absence of exogenous growth factors. Concomitant with the loss of growth factor dependence in culture, the Ab-MuLV-infected cell lines become tumorigenic in syngeneic mice. This secondary level of transformation is Ab-MuLV specific. Fr-MuLV-immortalized myeloid cell lines superinfected with Harvey murine sarcoma virus (Ha-MuSV) or amphotropic virus remain dependent on WEHI-3 CM for growth in vitro and are not tumorigenic in vivo. Neither Ab-MuLV- nor Ha-MuSV-infected normal mouse myeloid cell cultures produce growth factor-independent or tumorigenic cell lines. We conclude that at least two genetic events are needed to convert a murine myeloid precursor into a tumorigenic cell line. The first event occurs in Fr-MuLV infected mice, generating cells that are growth factor dependent but immortal in vitro. The second event, which can be accomplished by Ab-MuLV infection, converts these immortal myeloid precursors into growth factor-independent and tumorigenic cells. PMID- 2987919 TI - Chromosomal assignments of the genes coding for human types II, III, and IV collagen: a dispersed gene family. AB - The human type II collagen gene, COL2A1, has been assigned to chromosome 12, the type III gene, COL3A1, to chromosome 2, and one of the type IV genes, COL4A1, to chromosome 13. These assignments were made by using cloned genes as probes on Southern blots of DNA from a panel of mouse/human somatic cell hybrids. The two genes of type I collagen, COL1A1 and COL2A1, have been mapped previously to chromosomes 17 and 7, respectively. This family of conserved genes seems therefore to be dispersed throughout the genome. PMID- 2987921 TI - Genes for respiratory chain proteins and ribosomal RNAs are present on a 16 kilobase-pair DNA species from Chlamydomonas reinhardtii mitochondria. AB - We have used heterologous hybridization and DNA sequence analysis to determine whether the 16-kilobase-pair (kbp) DNA from Chlamydomonas reinhardtii mitochondria is the functional equivalent of mtDNA in other eukaryotes. Restriction fragments corresponding to a continuous internal stretch spanning 75% of the 16-kbp DNA have been cloned and mapped, and regions hybridizing with probes specific for the cytochrome oxidase subunit I [CytOx I (acronym COI)] and apocytochrome b (Cyt b) genes of yeast and the mitochondrial 26S and 18S rRNA genes of wheat have been identified. Sequence analysis has verified the presence of CytOx I and the large and small subunit rRNA genes in the C. reinhardtii 16 kbp DNA. In the region of the 16-kbp DNA corresponding to exon 4 in the yeast CytOx I gene, the derived amino acid sequence is 61% and 63% identical with the CytOx I amino acid sequences of yeast and human mitochondria, respectively. Notably, tryptophan is specified by TGG rather than by TGA in this section of the C. reinhardtii CytOx I gene. A probe from the CytOx I region of the 16-kbp DNA hybridizes only with this 16-kbp DNA in Southern blots of total cellular DNA from C. reinhardtii but with a larger DNA species in the total cellular DNA of C. moewusii and C. eugametos--two species that lack a 16-kbp DNA. These observations provide evidence that C. reinhardtii 16-kbp DNA comprises at least part of the mitochondrial genome of this organism and that a homologous DNA exists in other species of Chlamydomonas. PMID- 2987920 TI - A systematic study of HLA class II-beta DNA restriction fragments in insulin dependent diabetes mellitus. AB - DNA restriction fragments of the genes encoding HLA class II-beta antigens were compared in 34 patients with insulin-dependent diabetes mellitus and 34 HLA-DR matched healthy individuals. Ninety-three fragments, determined by six restriction enzymes (EcoRI, EcoRV, HindIII, BamHI, Pvu II, and Taq I), were analyzed: (i) A DR Taq I 12.7-kilobase-pair fragment might be a marker for the extended haplotype HLA-B8, DR3. (ii) In controls, DR4 haplotypes are associated with two distinct clusters of DQ restriction fragments (DQR4 and DQR5). Almost all (94%) DR4 patients belong to the DQR4 and not to the DQR5 cluster. This suggests that, among HLA-DR4 haplotypes, only DQR4 haplotypes are involved in susceptibility to insulin-dependent diabetes mellitus. (iii) A DR Taq I 14.5 kilobase-pair fragment was found to be strongly associated with DQR4, mainly in DR3/DR4 heterozygous patients (P = 5 X 10(-4). However, these results must be interpreted with caution, taking into account the high number of statistical tests performed. PMID- 2987922 TI - Relative rates of homologous and nonhomologous recombination in transfected DNA. AB - Both homologous and nonhomologous recombination events occur at high efficiency in DNA molecules transfected into mammalian cells. Both types of recombination occur with similar overall efficiencies, as measured by an endpoint assay, but their relative rates are unknown. In this communication, we measure the relative rates of homologous and nonhomologous recombination in DNA transfected into monkey cells. This measurement is made by using a linear simian virus 40 genome that contains a 131-base-pair duplication at its termini. Once inside the cell, this molecule must circularize to initiate lytic infection. Circularization can occur either by direct, nonhomologous end-joining or by homologous recombination within the duplicated region. Although the products of the two recombination pathways are different, they are equally infectious. Since homologous and nonhomologous recombination processes are competing for the same substrate, the relative amounts of the products of each pathway should reflect the relative rates of homologous and nonhomologous recombination. Analysis of individual recombinant genomes from 164 plaques indicates that the rate of circularization by nonhomologous recombination is 2- to 3-fold higher than the rate of homologous recombination. The assay system described here may prove to be useful for testing procedures designed to influence the relative rates of homologous and nonhomologous recombination. PMID- 2987923 TI - Analysis of DNA haplotypes suggests a genetic predisposition to trisomy 21 associated with DNA sequences on chromosome 21. AB - To test the hypothesis that there is a genetic predisposition to nondisjunction and trisomy 21 associated with DNA sequences on chromosome 21, we used DNA polymorphism haplotypes for chromosomes 21 to examine the distribution of different chromosomes 21 in Down syndrome and control families from the same ethnic group. The chromosomes 21 from 20 Greek families with a Down syndrome child and 27 control Greek families have been examined for DNA polymorphism haplotypes by using four common polymorphic sites adjacent to two closely linked single-copy DNA sequences (namely pW228C and pW236B), which map somewhere near the proximal long arm of chromosome 21. Three haplotypes, +, +---, and - with respective frequencies of 43/108, 24/108, and 23/108, account for the majority of chromosomes 21 in the control families. However, haplotype - was found to be much more commonly associated with chromosomes 21 that underwent nondisjunction in the Down syndrome families (frequency of 21/50; X2 for the two distributions is 9.550; P = 0.023; degrees of freedom, 3). The two populations (control and trisomic families) did not differ in the distribution of haplotypes for two DNA polymorphisms on chromosome 17. The data from this initial study suggest that the chromosome 21, which is marked in Greeks with haplotype - for the four above described polymorphic sites, is found more commonly in chromosomes that participate in nondisjunction than in controls. We propose an increased tendency for nondisjunction due to DNA sequences associated with a subset of chromosomes 21 bearing this haplotype. PMID- 2987924 TI - Generation of a dominant 8-MDa deletion at the left terminus of vaccinia virus DNA. AB - Vaccinia virus mutants were obtained in high frequency from mouse Friend erythroleukemia (FEL) cells persistently infected with this virus, which contains a large (122-MDa) DNA. During long-term cell passages viral particles with deletions of the DNA are generated in FEL cells. These mutants have a major 8-MDa deletion starting between 2.2 and 3.2 MDa from the left terminus of the viral genome. More than half of the left end terminal repetition is deleted. These mutants have reduced infectivity compared to wild-type virus. The ease with which vaccinia virus mutants are obtained in FEL cells should provide a suitable system for generating mutants with other poxvirus and permit study of the genetic basis of virulence for this group of viruses. PMID- 2987925 TI - Isolation of a gene enhancer within an amplified inverted duplication after "expression selection". AB - We have attempted to isolate and identify cellular expression sequences from F9 teratocarcinoma DNA by utilizing their ability to reactivate a selectable gene devoid of its own expression sequences (expression selection). Restriction nuclease-digested F9 cellular DNA was ligated to a polyoma virus (Py) DNA fragment which contains an intact transforming region but is incapable of inducing transformation because it lacks the viral 5' enhancer sequence. The ligation mixture was used to transfect Rat-1 cells and a transformed cell line, 3B, was isolated. The 3B cell line contained a single type of Py DNA insert, which was molecularly cloned as an 18-kilobase BglII fragment. A weak cellular enhancer was identified in a 4.7-kilobase BamHI fragment upstream from the Py sequences. Both the Py DNA and the enhancer sequences were found to be present in an inverted duplication in the 3B clone. The presence of this structure in 3B genomic DNA was confirmed by the analysis of selectively isolated inverted duplicated sequences, and the structure was found to be at least 22 kilobases long. In the 3B cell line, the inverted duplicated sequences containing the Py and enhancer sequences are quite stable and are amplified 20- to 40-fold. The strongly transformed phenotype of the 3B cells may be a result of this amplification. The formation of inverted duplications as a part of the amplification mechanism as well as a general strategy for the cloning of inverted duplicated (amplified) sequences is discussed. PMID- 2987926 TI - Expression of a second Epstein-Barr virus-determined nuclear antigen in mouse cells after gene transfer with a cloned fragment of the viral genome. AB - Large Epstein-Barr virus (EBV) DNA restriction fragments corresponding to regions transcribed in transformed, proliferating cells were cloned in a cosmid derivative of the dominant-acting selection vector pSV2-gpt. Recombinant vectors carrying the EcoRI A fragment of EBV DNA were modified in the region corresponding to the deletion of the virion DNA in the non-transforming viral substrain P3HR-1, to create a series of recombinants lacking parts of this region. The recombinant vectors were introduced into 3T3 mouse fibroblasts under selective conditions, and resistant clones shown to contain EBV DNA sequences were analyzed for the expression of EBV-related antigens detectable by direct, indirect, and anticomplement immunofluorescence techniques. Cells that contained the BamHI K fragment expressed the EBV-determined nuclear antigen (EBNA) as expected. Cells transfected with recombinant vectors containing the BamHI W, Y, and H fragment part of the EcoRI A fragment also express a nuclear antigen detectable with certain anti-EBNA-positive human sera in anticomplement immunofluorescence tests. The BamHI WYH-induced EBNA polypeptide is similar in size to the EBNA2 polypeptide in Raji cells, as shown by gel electrophoresis and immunoblotting. The antigen is not detected in cells transfected with EcoRI A derived vectors in which the BamHI H fragment has been deleted or in cells transformed with vectors carrying the BamHI H fragment alone. Direct and indirect immunofluorescence did not reveal the presence of antigens associated with productive infection in any of the EBV DNA-transfected fibroblast clones. PMID- 2987927 TI - Nucleotide sequence and structure of the human apolipoprotein E gene. AB - The gene for human apolipoprotein E (apo-E) was selected from a library of cloned genomic DNA by screening with a specific cDNA hybridization probe, and its structure was characterized. The complete nucleotide sequence of the gene as well as 856 nucleotides of the 5' flanking region and 629 nucleotides of the 3' flanking region were determined. Analysis of the sequence showed that the mRNA encoding region of the apo-E gene consists of four exons separated by three introns. In comparison to the structure of the mRNA, the introns are located in the 5' noncoding region, in the codon for glycine at position -4 of the signal peptide region, and in the codon for arginine at position +61 of the mature protein. The overall lengths of the apo-E gene and its corresponding mRNA are 3597 and 1163 nucleotides, respectively; a mature plasma protein of 299 amino acids is produced by this gene. Examination of the 5' terminus of the gene by S1 nuclease mapping shows apparent multiple transcription initiation sites. The proximal 5' flanking region contains a "TATA box" element as well as two nearby inverted repeat elements. In addition, there are four Alu family sequences associated with the apo-E gene: an Alu sequence located near each end of the gene and two Alu sequences located in the second intron. This knowledge of the structure permits a molecular approach to characterizing the regulation of the apo-E gene. PMID- 2987928 TI - T-cell receptor gene rearrangements as markers of lineage and clonality in T-cell neoplasms. AB - Ig gene rearrangements represent markers of lineage, clonality, and differentiation of B cells, allowing a molecular diagnosis and immunogenotypic classification of B-cell neoplasms. We sought to apply a similar approach to the study of T-cell populations by analyzing rearrangements of the T-cell receptor beta-chain (T beta) gene. Our analysis, by Southern blotting hybridization using T beta-specific probes of DNAs from polyclonal T cells and from 12 T-cell tumors, indicates that T beta gene rearrangement patterns can be used as markers of (i) lineage, allowing the identification of polyclonal T-cell populations, and (ii) clonality, allowing the detection of monoclonal T-cell tumors. In addition, our data indicate that T beta gene rearrangements represent early and general markers of T-cell differentiation since they are detectable in histologically different tumors at all stages of T-cell development. The ability to determine lineage, clonality, and stage of differentiation has significant implications for future experimental and clinical studies on normal and neoplastic T cells. PMID- 2987929 TI - The postsynaptic density: a possible role in long-lasting effects in the central nervous system. AB - A theory is proposed that biochemical changes at the synapse that occur as a result of stimulation of specific neuronal circuits can lead to long-term changes only if alterations occur in synaptic structures in these circuits. The main synaptic structure that is thought to undergo this alteration is the postsynaptic density (PSD). There are many reports in the literature of overall structural changes at the synapse, including the PSD, resulting from various neuronal stimuli. These structural changes are here envisaged to include those of concentration and conformation of PSD proteins, changes that could alter the neural physiology of dendritic spines and even that of the presynaptic terminal. PMID- 2987930 TI - Characterization of cultured rat oligodendrocytes proliferating in a serum-free, chemically defined medium. AB - A serumless, chemically defined medium has been developed for the culture of oligodendrocytes isolated from primary neonatal rat cerebral cultures. Combined together, insulin, transferrin, and fibroblast growth factor synergistically induced an essentially homogenous population (95-98%) of cells expressing glycerol-3-phosphate dehydrogenase (EC 1.1.1.8) activity to undergo cell division. Proliferating cels were characterized by several criteria: (i) ultrastructural analysis by transmission electron microscopy identified the cell type as an oligodendrocyte; (ii) biochemical assays showed expression of three oligodendrocyte biochemical markers, induction of both glycerol phosphate dehydrogenase and lactate dehydrogenase (EC 1.1.1.27), and presence of 2',3' cyclic nucleotide 3'-phosphodiesterase (EC 3.1.4.37); and (iii) immunocytochemical staining showed cultures to be 95-98% positive for glycerol phosphate dehydrogenase, 90% for myelin basic protein, 60-70% for galactocerebroside, and 70% for A2B5. Few cells (less than 5%) stained positive for glial fibrillary acidic protein, and none were detected positive for fibronectin. PMID- 2987931 TI - Ultrashort-loop positive feedback of corticotropin (ACTH)-releasing factor to enhance ACTH release in stress. AB - Previous experiments have shown that intraventricular injection of ovine corticotropin (ACTH)-releasing factor (oCRF) in doses too low to elevate plasma ACTH by direct action on the pituitary does not lower plasma ACTH, suggesting that the peptide lacks a negative ultrashort-loop feedback action to suppress its own release under resting conditions. The present study was performed to determine whether oCRF has any action to alter CRF release in stress. The peptide was injected into the third ventricle or external jugular vein of freely moving ovariectomized female rats 5 min prior to application of ether stress. When oCRF was injected into the third ventricle in doses of 500 pg (0.1 pmol) or less, there was no significant alteration in plasma ACTH prior to ether stress; however, there was a significantly enhanced increase in plasma ACTH 2 and 5 min after ether stress applied 5 min after intraventricular injection of oCRF at doses of 50 (0.01 pmol) or 150 pg (0.03 pmol). These results suggest that the peptide acts on structures adjacent to the third ventricle to augment stress induced CRF release. To rule out the possibility that the sensitivity of the pituitary itself to CRF increases dramatically following stress, 10 or 100 ng of oCRF was injected i.v. These doses produced a significant dose-related increase in plasma ACTH at 2, 5, or 15 min. In other groups receiving the same doses of oCRF and ether stressed 5 min later, plasma ACTH was significantly higher 2 or 5 min after ether stress when compared with plasma ACTH in ether-stressed saline injected animals. However, in contrast with the results of intraventricular injection of oCRF, the release of ACTH was no greater than that obtained by summing the independent effects of exogenous oCRF and the CRF released by stress. We conclude that CRF may have a positive ultrashort-loop feedback action to enhance stress-induced ACTH release and that this enhancement is not due to increased sensitivity of anterior pituitary corticotrophs to CRF. PMID- 2987932 TI - Coding sequences for vasoactive intestinal peptide and PHM-27 peptide are located on two adjacent exons in the human genome. AB - The human precursor gene for vasoactive intestinal peptide (VIP) and PHM-27, a peptide that has an NH2-terminal histidine and COOH-terminal methionine amide and is closely related in sequence and activity to VIP, was detected with synthetic oligodeoxynucleotide probes. These specific hybridization segments were constructed according to the neuroblastoma VIP cDNA sequence and contained up to 39 bases. The gene structure was partly deduced by hybridization to synthetic oligodeoxynucleotide probes and partly by direct chemical nucleotide sequencing. Four exons were discovered thus far; among them are two short exons separated by a 0.75-kilobase DNA stretch, one encoding PHM-27 and the second encoding VIP (exons 1 and 2). Each of these two exons encodes both the hormone amino acid residues as well as the post-translational processing signal sequences. The 3' splice sites of the two exons contain an identical stretch of nine nucleotides. At the cDNA level, the 3' splice sites contain the same stretch of six nucleotides, which are identically spliced. The occurrence of VIP and PHM-27 coding sequences on two separate exons of the human genome and the homology of their 3' splice site may allow alternative RNA processing as discussed below. PMID- 2987933 TI - Initiation of enzymatic replication at the origin of the Escherichia coli chromosome: contributions of RNA polymerase and primase. AB - Replication of plasmids that depend on the 245-base-pair origin of the Escherichia coli chromosome (oriC) requires many purified proteins that (i) direct initiation to oriC (e.g., dnaA protein), (ii) influence initiations elsewhere (e.g., auxiliary proteins), and (iii) prime and extend DNA chains (e.g., priming and synthesis proteins). For the RNA priming and initiation of new DNA chains, the requirements for both primase and RNA polymerase (RNA pol) [Kaguni, J. M. & Kornberg, A. (1984) Cell 38, 183-190] have been further analyzed. Depending on the levels of auxiliary proteins (topoisomerase I and protein HU), three priming systems can operate: primase alone, RNA pol alone, or both combined. At low levels of auxiliary proteins, primase alone sustains an effective priming system. At higher levels, primase action is blocked, but RNA pol alone can initiate replication, albeit feebly; at these high levels of auxiliary proteins, primase and RNA pol act synergistically. When RNA pol is stalled by an inhibitor or lack of a ribonucleoside triphosphate, primase action is also inhibited. Based on these and other data [van der Ende, A., Baker, T. A., Ogawa, T. & Kornberg, A. (1985) Proc. Natl. Acad. Sci. USA 82, in press], RNA pol can counteract inhibition by auxiliary proteins and thus activate the origin for the priming by primase of the leading strand of the replication fork. PMID- 2987934 TI - Overproduction of the protein product of a nonselected foreign gene carried by an adenovirus vector. AB - We have constructed a recombinant adenovirus that carries the herpes simplex virus type I gene for thymidine kinase (EC 2.7.1.21) and expresses thymidine kinase under control of adenovirus major late promoter. A DNA fragment carrying thymidine kinase coding sequences but lacking the thymidine kinase promoter was sandwiched between a piece of adenoviral DNA and simian virus 40 early DNA on a plasmid. The aligned fragment was then inserted into the adenoviral genome, replacing internal adenoviral DNA. Hybrid viruses carrying the thymidine kinase gene were obtained by selecting for viruses that express simian virus 40 tumor antigen (T antigen) in monkey cells. The thymidine kinase gene was positioned in the third segment of the adenovirus tripartite leader downstream from the major late promoter by in vivo DNA recombination between the duplicated adenoviral sequences present in the plasmid insert and the viral vector. Levels of thymidine kinase activity in human or monkey cells infected with this hybrid virus were several times higher than in cells infected with herpes simplex virus. Infected cells produced thymidine kinase protein at very high levels, similar to those found for adenovirus late major capsid proteins. The thymidine kinase protein represented 10% of the newly synthesized protein in late infected cells and accumulated to represent 1% of total cell protein under optimal conditions. This vector system offers a procedure by which a variety of gene products that are biologically active and properly modified can be produced at high levels in mammalian cells. PMID- 2987935 TI - Three-dimensional structure of ubiquitin at 2.8 A resolution. AB - The three-dimensional structure of ubiquitin has been determined at 2.8 A resolution. X-ray diffraction data for the native protein and derivatives were collected with an automated diffractometer. Phases were obtained by use of a single isomorphous mercuric acetate derivative. The molecule contains a pronounced hydrophobic core. Prominent secondary structural features include three and one-half turns of alpha-helix, a mixed beta-sheet that contains four strands, and seven reverse turns. The histidine, tyrosine, and two phenylalanine residues are located on the surface of the molecule. PMID- 2987936 TI - Thyroid hormones increase Na+-H+ exchange activity in renal brush border membranes. AB - Na+-H+ exchange activity, i.e., amiloride-sensitive Na+ and H+ flux, in renal proximal tubule brush border (luminal) membrane vesicles was increased in the hyperthyroid rat and decreased in the hypothyroid rat, relative to the euthyroid animal. A positive correlation was found between Na+-H+ exchange activity and serum concentrations of thyroxine (T4) and triiodothyronine (T3). The thyroid status of the animal did not alter amiloride-insensitive Na+ uptake. The rate of passive pH gradient dissipation was higher in membrane vesicles from hyperthyroid rats compared to the rate in vesicles from hypothyroid animals, a result which would tend to limit the increase in Na+ uptake in vesicles from hyperthyroid animals. Na+-dependent phosphate uptake was increased in membrane vesicles from hyperthyroid rats; Na+-dependent D-glucose and L-proline uptakes were not changed by the thyroid status of the animal. The effect of thyroid hormones in increasing the uptake of Na+ in the brush border membrane vesicle is consistent with the action of the hormones in enhancing renal Na+ reabsorption. Further, the regulation of transtubular Na+ flux has now been shown to be concomitant with modulation of the entry of Na+ into the tubular cell across its luminal membrane, mediated by the exchange reaction, and with the previously reported control of the pumping of Na+ out of the cell across its basolateral membrane, mediated by the Na+,K+-ATPase. PMID- 2987937 TI - Monomers through trimers of large tumor antigen bind in region I and monomers through tetramers bind in region II of simian virus 40 origin of replication DNA as stable structures in solution. AB - Large tumor (T) antigen and its bound multimeric states are positioned by scanning transmission electron microscopy (STEM) within a few base pairs at control sequences of the simian virus 40 DNA origin of replication region. Proximal and distal edge positions for each multimer group match the end positions of previously mapped fragments protected from DNase cleavage. Since chance correspondence is shown to be extremely unlikely, STEM mass measurements, obtained concurrently with STEM map positions, indicate that the DNase fragments arise from bound monomers, dimers, trimers, and tetramers in binding region II and monomers, dimers, and trimers in binding region I. Simultaneous binding of seven monomer-equivalent masses is observed, three in region I and four in region II, with an ordered and interpretable mass distribution in the plane of the foil. Although this observation does not prove that the six G-A-G-G-C and one T-A-G-G-C sequences, similarly distributed, function as recognition sequences for T-antigen monomer, it provides strong support for such a model. The stable existence in solution of low-and intermediate-mass structures, observed at lower T-antigen concentrations, suggests a role as assembly intermediates. PMID- 2987938 TI - Expression of human choriogonadotropin in monkey cells using a single simian virus 40 vector. AB - We have inserted the cDNAs coding for both polypeptide subunits, alpha and beta, of human choriogonadotropin (hCG) into a single simian virus 40 expression vector in such a way that they replace the viral VP2 and VP1 coding sequences, respectively. Monkey cells infected with this virus and the appropriate helper virus produce dimeric hCG. hCG produced in this system was shown to chromatograph identically to standard hCG preparations on gel filtration and to be biologically active in the mouse uterine weight assay. PMID- 2987939 TI - Application of the principle of linked functions to ATP-driven ion pumps: kinetics of activation by ATP. AB - If a ligand binds with unequal affinity to two distinct states of a protein, then the equilibrium between the two states becomes a function of the concentration of the ligand. A necessary consequence is that the ligand must also affect the forward and/or reverse rate constants for transition between the two states. For an enzyme or transport protein with such a transition as a slow step in the catalytic cycle, the overall rate also becomes a function of ligand concentration. These conclusions are independent of whether or not the ligand is a direct participant in the reaction. If it is a direct participant, then the kinetic effect arising from the principle of linked functions is distinct from the direct catalytic effect. These principles suffice to account for the biphasic response of the hydrolytic activity of ATP-driven ion pumps to the concentration of ATP, without the need to invoke more than one ATP binding site per catalytic center. PMID- 2987940 TI - A truncated immunoglobulin epsilon pseudogene is found in gorilla and man but not in chimpanzee. AB - Molecular genetic analyses of the young pseudogenes of the immunoglobulin C epsilon genes were carried out to obtain qualitative evidence for the phylogenetic branching pattern of hominoid primates. We found that Old World monkeys had two C epsilon genes, one of which was processed. Among the hominoids examined only the gorilla and human genomes contained three C epsilon genes: an active, a truncated, and a processed gene. Other hominoids so far examined, including chimpanzee, contained two C epsilon genes: one active and the other processed. These results suggest that the processed C epsilon pseudogene was generated before the divergence between Old World monkeys and hominoids and that the gorilla is more closely related to man than the chimpanzee is, unless the chimpanzee has lost the C epsilon 2 gene after the divergence of this species. PMID- 2987941 TI - Processing of donor DNA during Haemophilus influenzae transformation: analysis using a model plasmid system. AB - A plasmid system was used to investigate the processing of donor DNA during transformation of competent Haemophilus influenzae. Using biochemical and genetic methods, we have determined that portion of a donor plasmid molecule that, on average, can become integrated into a homologous recipient plasmid during transformation. Our results show that (i) transformation efficiency decreases linearly with donor DNA length over the range of 11 to 3.5 kilobase pairs, (ii) transformation efficiency decreases exponentially with size for donor molecules less than 3.5 kilobase pairs in length, and (iii) 5'-end label, but not 3'-end label, can be specifically incorporated into the resident homologous region. We present a model for donor processing during entry that encompasses and explains these observations. PMID- 2987942 TI - Deletion mapping of HLA and chromosome 6p genes. AB - We have analyzed a set of heterozygous mutants with deletions that encompass parts of HLA and surrounding regions of chromosome 6p. By a combination of Southern blotting, serologic, enzymatic, and cytogenetic analyses, we have ordered eight independent deletion break points into a sequence that divides chromosome 6p into six regions. The deletion mutants have been used in conjunction with the Southern blot technique to map HLA and other 6p gene probes into those regions. On the basis of these and other data we propose a genetic and physical map of HLA and surrounding regions of chromosome 6p. We find that for HLA probes, most of which hybridize with more than one gene, the multiple copies recognized by single probes map to single regions. Any chromosome 6p gene can now be regionally mapped by using these mutants. PMID- 2987943 TI - Activation of the c-myc oncogene by the immunoglobulin heavy-chain gene enhancer after multiple switch region-mediated chromosome rearrangements in a murine plasmacytoma. AB - Presented is a detailed molecular analysis of the rearranged c-myc oncogene from ABPC45, an unusual plasmacytoma that was originally classified as translocation negative. Previous data obtained by high-resolution chromosome banding suggested that this tumor was a member of a small group distinguished by the absence of rcpt (12;15) or (6;15) and further characterized by a band deletion near the c myc locus on chromosome 15. However, genomic Southern blotting and analysis of the cloned oncogene in the present study reveal that (i) chromosome 12 sequences lie 365 base pairs 5' of the rearranged c-myc; (ii) this DNA consists of immunoglobulin alpha switch region and 5' immunoglobulin mu switch region sequences that are rearranged in an aberrant fashion; and (iii) the immunoglobulin heavy-chain gene enhancer element now resides approximately 2.5 kilobase pairs 5' of c-myc. We infer from these and other data that the rearrangement of c-myc in ABPC45 occurred via a multistep switch region-mediated process and that a reciprocal translocation has indeed taken place. Unlike many other plasmacytomas, this event did not interrupt the normal c-myc transcription unit. Rather, disruption of gene regulation is manifested in part by a change in relative usage of the two promoters normally used by the unrearranged gene. In contrast to several of its counterparts in Burkitt lymphomas, DNA sequence analysis of the translocated c-myc gene of ABPC45 reveals that it has not acquired point mutations in the noncoding first exon. These results strongly imply that a cis-acting regulatory element normally located 5' of exon 1 is lost and that heavy-chain constant region or enhancer sequences exert similar cis effects on translocated c-myc loci. PMID- 2987944 TI - Cloning of cDNAs for human aldehyde dehydrogenases 1 and 2. AB - Partial cDNA clones encoding human cytosolic aldehyde dehydrogenase (ALDH1) and mitochondrial aldehyde dehydrogenase (ALDH2) were isolated from a human liver cDNA library constructed in phage lambda gt11. The expression library was screened by using rabbit antibodies against ALDH1 and ALDH2. Positive clones thus obtained were subsequently screened with mixed synthetic oligonucleotides compatible with peptide sequences of ALDH1 and ALDH2. One of the positive clones for ALDH1 contained an insertion of 1.6 kilobase pairs (kbp). The insert encoded 340 amino acid residues and had a 3' noncoding region of 538 bp and a poly(A) segment. The amino acid sequence deduced from the cDNA sequence coincided with the reported amino acid sequence of human ALDH1 [Hempel, J., von Bahr-Lindstrom, H. & Jornvall, H. (1984) Eur. J. Biochem. 141, 21-35], except that valine at position 161 in the previous amino acid sequence study was found to be isoleucine in the deduced sequence. Since the amino acid sequence of ALDH2 was unknown, 33 tryptic peptides of human ALDH2 were isolated and sequenced. Based on the amino acid sequence data thus obtained, a mixed oligonucleotide probe was prepared. Two positive clones, lambda ALDH2-21 and lambda ALDH2-36, contained the same insert of 1.2 kbp. Another clone, lambda ALDH2-22, contained an insert of 1.3 kbp. These two inserts contained an overlap region of 0.9 kbp. The combined cDNA contained a sequence that encodes 399 amino acid residues, a chain-termination codon, a 3' untranslated region of 403 bp, and a poly(A) segment. The deduced amino acid sequence was compatible with the amino acid sequences of the tryptic peptides. The degree of homology between human ALDH1 and ALDH2 is 66% for the coding regions of their cDNAs and 69% at the protein level. No significant homology was found in their 3' untranslated regions. PMID- 2987945 TI - Homologous recombination between a defective virus and a chromosomal sequence in mammalian cells. AB - Replacement of the early region of simian virus 40 results in virus that cannot replicate in a normal host, CV-1 cells, but can replicate in COS cells, a derivative of CV-1 cells that constitutively express simian virus 40 tumor antigen (T antigen). However, passage of such an early replacement simian virus 40 mutant in COS cells results in the emergence of virus that can propagate in CV 1 cells. Analysis of this virus revealed that the mutant rescued the integrated T antigen gene from the COS cell genome. Comparison of the sequence of the recovered virus with that of the viral DNA resident in COS cells (strain 776) and the mutant used in our studies (derived from strain 777) proves that the mutant virus acquired the T-antigen gene from the COS cell chromosome via homologous recombination. Most probably this process was mediated by a direct genetic exchange. PMID- 2987946 TI - The alpha-spectrin gene is on chromosome 1 in mouse and man. AB - By using alpha-spectrin cDNA clones of murine and human origin and somatic cell hybrids segregating either mouse or human chromosomes, the gene for alpha spectrin has been mapped to chromosome 1 in both species. This assignment of the mouse alpha-spectrin gene to mouse chromosome 1 by DNA hybridization strengthens the previous identification of the alpha-spectrin locus in mouse with the sph locus, which previously was mapped by linkage analysis to mouse chromosome 1, distal to the Pep-3 locus. By in situ hybridization to human metaphase chromosomes, the human alpha-spectrin gene has been localized to 1q22-1q25; interestingly, the locus for a non-Rh-linked form of elliptocytosis has been provisionally mapped to band 1q2 by family linkage studies. PMID- 2987947 TI - Segregation of polymorphic T-cell receptor genes in human families. AB - Polymorphism in the genes encoding the constant (C) region of the beta chain of the T-cell antigen receptor (CT beta, also called C beta) has been detected by molecular genotyping analyses. In initial screenings, a panel of restriction endonucleases was used to digest DNA samples from two individuals; the digested samples were subjected to Southern blot analyses using a CT beta probe. The enzyme Bgl II revealed restriction-fragment-length polymorphism in these samples and was subsequently used to test 59 individual members of eight different families. Polymorphic fragments detected in six of the families could be used to follow the segregation of T-cell receptor genes; in many cases maternal and/or paternal haplotypes could be assigned. All members of two additional families displayed a single CT beta hybridizing fragment. In one family the DNA sample from one of the children lacked an expected Bgl II restriction fragment. On the basis of analyses with other restriction enzymes, the most likely explanation is that the lymphoblastoid B-cell line used as a source of genomic DNA for this individual had rearranged or altered CT beta genes. Restriction-fragment-length polymorphisms used to discriminate CT beta haplotypes in families provide useful markers that will facilitate linkage studies and genetic analyses of T-cell function. PMID- 2987948 TI - The C1q subunit of the first component of complement binds to laminin: a mechanism for the deposition and retention of immune complexes in basement membrane. AB - The C1q subunit of complement component C1 is known to bind to immune complexes, which often are deposited in basement membrane. We investigated the possibility that this deposition is a result of binding to laminin, a large basement membrane glycoprotein. C1q showed saturable binding to immobilized laminin; this binding was increased at reduced ionic strength. Intact C1 did not bind laminin. A ternary complex was formed by laminin, C1q, and aggregated IgG. This complex formation was dependent on and proportional to the amount of C1q bound to the aggregated IgG. Binding of laminin to C1q occurred with a Kd of 2 nM and was stronger than the binding of C1q to fibronectin. Preliminary data, including electron micrographs of rotary-shadowed preparations, suggest that laminin binds to the collagen-like tail of C1q. Electron microscopy localized the site of interaction with C1q to a short arm of laminin. Since laminin is found only in basement membranes, the interaction between laminin and C1q could be involved in the deposition and retention of immune complexes in these structures. PMID- 2987949 TI - Leukotriene C4 as a mediator of luteinizing hormone release from rat anterior pituitary cells. AB - This study demonstrates that leukotriene C4, at concentrations in the picomolar range, released luteinizing hormone (LH) but not growth hormone (GH) from dispersed rat anterior pituitary cells. Leukotriene B4, another lipoxygenase pathway product of arachidonic acid, had no effect on LH or GH release. The stimulatory effect of leukotriene C4 could be seen after 0.5 but not after 3 hr of incubation. This was in contrast to the dose-dependent LH-releasing hormone (LHRH)-induced LH release that was not measurable after 0.5 hr but was fully established after incubation for 3 hr. Furthermore, the LH-releasing ability of leukotriene C4 was blocked in the presence of high doses of LHRH. The immunohistochemical analysis revealed leukotriene C4-immunoreactive fibers at all levels of the median eminence, mainly in the lateral parts. These fibers exhibited a marked overlap distribution with LHRH-immunoreactive fibers and elution-restaining experiments revealed identity of at least a large proportion of the leukotriene C4- and LHRH-immunoreactive fibers. Furthermore, cell bodies in the preoptic area contained both leukotriene C4- and LHRH-like immunoreactivities, suggesting localization of these two compounds in the same neurons. PMID- 2987950 TI - Frequency of molecular alterations affecting ras protooncogenes in human urinary tract tumors. AB - Members of the ras gene family are activated as oncogenes in many different human cancers. To systematically determine the frequency at which such genes might be involved in the neoplastic process affecting a specific target tissue, urothelial cells, we surveyed a large series of urinary tract tumors for ras oncogenes by DNA transfection and by molecular genetic analysis. Harvey (Ha)-ras oncogenes were detected in 2 of 38 tumors by transfection, molecularly cloned in biologically active form, and shown to contain single base changes at codon 61 leading to substitutions of arginine and leucine, respectively, for glutamine at this position. One additional Ha-ras oncogene was identified in a bladder carcinoma by restriction polymorphisms at codon 12. In one of 21 tumors, we observed a 40-fold amplification of the Kirsten (Ki)-ras gene. No amplification of other ras genes was detected in any of the tumors analyzed. Our findings strengthen the conclusion that codons 12 and 61 are the major "hot spots" of ras oncogene activation and suggest that quantitative alterations in expression due to gene amplification may provide an alternative mechanism for ras gene activation in primary human tumors. PMID- 2987952 TI - Asbestos-associated chromosomal changes in human mesothelial cells. AB - Replicative cultures of human pleural mesothelial cells were established from noncancerous adult donors. The cells exhibited normal mesothelial cell characteristics including keratin, hyaluronic acid mucin, and long branched microvilli, and they retained the normal human karyotype until senescence. The mesothelial cells were 10 and 100 times more sensitive to the cytotoxic effects of asbestos fibers than normal human bronchial epithelial or fibroblastic cells, respectively. In addition, cultures of mesothelial cells that survived two cytotoxic exposures of amosite fibers were aneuploid with consistent specific chromosomal losses indicative of clonal origin. These aneuploid cells exhibit both altered growth control properties and a population doubling potential of greater than 50 divisions beyond the culture life span (30 doublings) of the control cells. PMID- 2987951 TI - Phorbol myristate acetate inhibits thrombin-stimulated Ca2+ mobilization and phosphatidylinositol 4,5-bisphosphate hydrolysis in human platelets. AB - The tumor-promoting phorbol diester 4 beta-phorbol 12-myristate 13-acetate (PMA) inhibited mobilization of intracellular Ca2+ in platelets by thrombin (also trypsin and 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphocholine). PMA was effective over the same concentration range that activates protein kinase C in intact platelets; IC50 vs. thrombin = 2 ng/ml, 3.4 nM: greater than 90% inhibition at 10 20 ng/ml. Suppression of thrombin-induced Ca2+ mobilization was evident within 30 sec of pretreatment with PMA and was essentially complete by 6-10 min at 10-20 ng of PMA per ml. Thrombin-induced secretion was initially accelerated in the presence of PMA, but after 1 min it was progressively inhibited when Ca2+ mobilization was depressed by greater than 60%. PMA did not inhibit Ca2+ mobilization or secretion caused by A23187. Thrombin-induced phosphatidylinositol 4,5-[32P]bisphosphate breakdown and [32P]phosphatidic acid production were also initially increased by PMA and then progressively depressed. Inhibition of thrombin-induced lipid metabolism required higher concentrations of PMA (IC50 = 10 ng/ml), and it was not overcome by A23187. 4 alpha-Phorbol 12,13-didecanoate, which lacks the ability to activate protein kinase C, did not inhibit any responses to thrombin. These results suggest that activation of protein kinase C, which initially fosters secretion and aggregation, may subsequently exert negative feedback on the receptor-mediated mobilization of intracellular Ca2+ and the hydrolysis of phosphatidylinositol 4,5-bisphosphate. PMID- 2987955 TI - Synaptic transmission from rods to bipolar cells in the tiger salamander retina. AB - Synaptic transmission between rods and depolarizing bipolar cells (DBC) was studied by using simultaneous recording techniques in the living retinal slice preparation. Current injection into the rod elicited a sign-inverting, sustained voltage change in the DBC. Voltage "tails" after the termination of a bright flash were observed in dark-adapted rods and DBC but not in cones. These simultaneously recorded voltage tails were used to isolate the rod input from the cone input and to study the input-output relation of the rod-DBC synapse. Within the voltage range between 0 and -10 mV from the rod dark potential (-39 +/- 1.2 mV), the input-output relation of the rod-DBC was approximately linear, with an estimated gain of about 3.7. PMID- 2987953 TI - Reduced in vivo mutagenesis by mutant herpes simplex DNA polymerase involves improved nucleotide selection. AB - We present evidence that mutation frequencies in a mammalian system can vary according to the replication fidelity of the DNA polymerase. We demonstrated previously that several derivatives of herpes simplex virus type 1 that encode polymerases resistant to various antiviral drugs (e.g., nucleotide analogues) also produce reduced numbers of spontaneous mutants. Here we show that the DNA polymerase from one antimutator virus exhibits enhanced replication fidelity. First, the antimutator virus showed a reduced response to known mutagens that promote base mispairing during DNA replication (N-methyl-N'-nitro-N nitrosoguanidine, 5-bromo-deoxyuridine). Second, purified DNA polymerase from the antimutator produced fewer replication errors in vitro, based on incorporation of mispaired nucleotides or analogues with abnormal sugar rings. We have investigated possible mechanisms for the enhanced fidelity of the antimutator polymerase. We show that the mutant enzyme has altered interactions with nucleoside triphosphates, as indicated by its resistance to nucleotide analogues and elevated Km values for normal nucleoside triphosphates. We present evidence against increased proofreading by an associated 3',5' exonuclease (as seen for T4 bacteriophage antimutator polymerases), based on nuclease levels in the mutant polymerase. We propose that reduced affinity of the polymerase for nucleoside triphosphates accounts for the antimutator phenotype by accentuating differences in base-pair stability, thus facilitating selection of correct nucleotides. PMID- 2987954 TI - Purification and characterization of a human T-lymphocyte-derived glial growth promoting factor. AB - Antigen- or lectin-stimulated T lymphocytes and human T-cell leukemia virus (HTLV)-infected cell lines secrete lymphokines that can influence the growth and function of a variety of cell types. We recently demonstrated that supernatants from the HTLV-II-infected Mo-T-cell line stimulate the proliferation of rat brain oligodendrocytes and astrocytes. We have now purified a glial growth-promoting factor (GGPF) from these supernatants. Purification from serum-free conditioned medium was accomplished by sequential concentration, ammonium sulfate precipitation, lentillectin affinity chromatography, gel filtration, and reversed phase high-performance liquid chromatography. GGPF is assayed by its ability to stimulate DNA synthesis in oligodendrocytes, as measured by [3H]thymidine uptake. The purified GGPF has an apparent Mr of 30,000 when analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis under nonreducing conditions. Under reducing conditions, however, GGPF appears as a single band of Mr 18,000. Both reduced and unreduced forms have biological activity, suggesting that GGPF exists in both a functional monomeric and dimeric form. Purified GGPF appears to be a biochemically and functionally distinct lymphokine. PMID- 2987956 TI - A sequence downstream of A-A-U-A-A-A is required for formation of simian virus 40 late mRNA 3' termini in frog oocytes. AB - The 3' terminus of simian virus 40 late mRNA is formed by nucleolytic cleavage of an mRNA precursor. In this report we show that efficient cleavage requires a sequence in the 3' flanking region, "downstream" of the highly conserved A-A-U-A A-A sequence. This sequence is not required for polyadenylylation itself or for accurately positioning the 3' terminus that is formed. PMID- 2987957 TI - Cloning of cDNA coding for peroxisomal acyl-CoA oxidase from the yeast Candida tropicalis pK233. AB - Candida tropicalis pK233 cells were grown with n-alkanes as carbon source to induce the synthesis of peroxisomal proteins and the proliferation of peroxisomes. Poly-(A)+ RNA was isolated and used to construct a cDNA library by insertion of double-stranded reverse transcripts into the Pst I site of pBR322 followed by cloning in Escherichia coli. Clones complementary to mRNAs induced by growth on alkanes were selected by differential DNA dot-blot analysis using [32P]cDNA reverse-transcribed from poly(A)+ RNA of glucose-grown cells (which contain few peroxisomes) or of alkane-grown cells. Among these clones, one containing a 1.7-kilobase insert coding for acyl-CoA oxidase (the first enzyme in the peroxisomal Beta-oxidation pathway) was identified by hybridization-selection translation and immunoprecipitation. By RNA blot analysis, the acyl-CoA oxidase mRNA was estimated to be approximately equal to 2.2 kilobases long, of which 2.1 kilobases are required to code for the approximately equal to 76-kDa protein. Since the mRNA is polyadenylylated, there appears to be little additional nontranslated region. Cell-free mRNA translation and RNA dot-blot hybridization analyses demonstrated that, whereas glucose-grown C. tropicalis contained little or no acyl-CoA oxidase mRNA, alkane-grown cells contained so much of this mRNA as to make acyl-CoA oxidase one of the major in vitro translation products. PMID- 2987958 TI - Evidence from two transformed cell lines that the phosphorylations of peptide tyrosine and phosphatidylinositol are catalyzed by different proteins. AB - Two transformed rodent cell lines (RS-1 and LSTRA) were studied in vitro to determine if their major protein tyrosine kinases catalyzed the phosphorylation of phosphatidylinositol (PtdIns), phosphatidylinositol 4-phosphate (PtdIns4P), or diacylglycerol. RS-1 cells, transformed by Rous sarcoma virus, contain high levels of pp60src; LSTRA cells, transformed by Moloney murine leukemia virus, contain a tyrosine kinase (pp56) that is the product of an unknown cellular gene. Rates of phosphorylation of peptide tyrosine were elevated more than 20-fold in RS-1 and LSTRA particulate fractions compared to fractions from suitable control cells (N2 and YAC-1), but there was not a proportional increase in rates of phosphorylation of PtdIns, PtdIns4P, or diacylglycerol. Heat (34 degrees C) completely inactivated the LSTRA tyrosine kinase, while it enhanced the phosphorylation of PtdIns and PtdIns4P and had no effect on the phosphorylation of diacylglycerol. PtdIns4P inhibited the phosphorylation of PtdIns but had no effect on tyrosine kinase activity. An antibody, raised against a peptide with a sequence homologous to the autophosphorylation site of pp60src, immunoprecipitated tyrosine kinase activity from RS-1 and LSTRA extracts but had no effect on PtdIns kinase or PtdIns4P kinase activity. These results provide evidence that the phosphorylations of tyrosine and PtdIns are catalyzed by different proteins. An additional observation was that a monoclonal antibody that binds to pp60src and pp56 removed PtdIns kinase as well as tyrosine kinase activity from RS-1 and LSTRA particulate extracts. This antibody also removed PtdIns kinase from N2 and YAC-1 extracts, in which tyrosine kinase activity was low or undetectable. Thus, the anti-pp60src monoclonal antibody may recognize the PtdIns kinase in addition to pp60src and pp56. PMID- 2987959 TI - Inositol cyclic phosphates are produced by cleavage of phosphatidylphosphoinositols (polyphosphoinositides) with purified sheep seminal vesicle phospholipase C enzymes. AB - Previous studies have shown that metabolism of phosphatidylinositol by phospholipase C produces a mixture of two water-soluble products: inositol 1 phosphate and inositol 1,2-(cyclic)phosphate. In the present study, we demonstrate that the water-soluble products of phosphatidylphosphoinositol (polyphosphoinositide) cleavage by purified ram seminal vesicle phospholipase C enzymes also contain cyclic phosphates. Inositol cyclic phosphates were detected by 18O labeling. In the presence of acid, cyclic phosphates are rapidly hydrolyzed to phosphomonoesters, and when the hydrolysis is carried out in H2 18O, the resultant phosphomonoesters will contain 18O. The 18O content of the phosphomonoesters was measured following alkaline phosphatase treatment and conversion of the inorganic phosphate to a volatile derivative for gas chromatography/mass spectrometry. Inositol cyclic phosphates were found in the phospholipase C cleavage products of all three phosphoinositides, but the ratio of cyclic to noncyclic product was found to decrease in the order phosphatidylinositol greater than phosphatidylinositol 4-phosphate greater than phosphatidylinositol 4,5-bisphosphate. The formation of myo-inositol 1,2(cyclic) 4-bisphosphate was further substantiated by anion-exchange HPLC of the water soluble products of [32P]phosphatidylinositol 4-phosphate metabolism by phospholipase C. Two peaks were detected one of which, on acid treatment, incorporated 18O from H2 18O into phosphate groups, consistent with this peak containing the cyclic phosphate product. These results suggest that polyphosphoinositide breakdown in stimulated cells may occur via a cyclic phosphate intermediate, as has been described for phosphatidylinositol. These cyclic phosphates contain a reactive bond that may play a role in phosphoinositide-derived signal transduction. PMID- 2987960 TI - Torsional stress induces an S1 nuclease-hypersensitive site within the promoter of the Xenopus laevis oocyte-type 5S RNA gene. AB - The internal promoter of the Xenopus laevis oocyte-type 5S RNA gene is preferentially cleaved by S1 and Bal-31 nucleases in plasmid DNA. S1 nuclease sensitivity is largely dependent on supercoiling; however, Bal-31 cleaves within the 5S RNA gene in linear as well as in supercoiled DNA. The S1 nuclease hypersensitive site is centered at position +48-52 of the gene at the 5' boundary of the promoter. A DNAase I-hypersensitive site is induced at this position upon binding of the transcription factor, TFIIIA, specific for the 5S RNA gene. The somatic-type 5S RNA gene promoter is not preferentially cleaved by S1 nuclease or Bal-31 nuclease in supercoiled DNA, nor does TFIIIA induce a DNase I site at position +50. This differential promoter response may be related to a 4-fold difference in TFIIIA affinity between the oocyte and somatic 5S RNA genes. PMID- 2987961 TI - Evidence that ribosomal protein S10 itself is a cellular component necessary for transcription antitermination by phage lambda N protein. AB - Bacteriophage lambda N gene product acts to modify host RNA polymerase allowing the formation of a termination-resistant transcription apparatus. Previous studies have demonstrated that the nusE71 mutation that has altered the ribosomal protein S10 prevents N action in vivo. Using a coupled transcription-translation system, we demonstrate here that purified S10 protein as well as the 30S ribosomal subunit is sufficient to restore N activity in the nusE mutant extract, allowing antitermination of Rho-dependent and Rho-independent terminators. This provides direct biochemical evidence that the S10 protein itself is one of the cellular components necessary for the formation of an antitermination apparatus. PMID- 2987962 TI - Platelet-derived growth factor mimics phorbol diester action on epidermal growth factor receptor phosphorylation at threonine-654. AB - Addition of platelet-derived growth factor (PDGF) to quiescent WI-38 human fetal lung fibroblasts mimics the effect of tumor-promoting phorbol diesters to inhibit the high-affinity binding of 125I-labeled epidermal growth factor (125I-EGF). PDGF, like phorbol diesters, was found to increase the phosphorylation state of EGF receptors immunoprecipitated from intact fibroblasts that were labeled to equilibrium with [32P]phosphate. Phosphoamino acid analysis of the EGF receptors indicated that both PDGF and phorbol diesters increased the level of [32P]phosphoserine and [32P]phosphothreonine. Phosphopeptide mapping of the EGF receptor demonstrated that PDGF increased the phosphorylation of several sites and induced the phosphorylation of a site that was not observed to be phosphorylated on EGF receptors isolated from control cells. This latter phosphorylation site on the EGF receptor was identified as threonine-654, previously shown to be phosphorylated in response to phorbol diesters in intact cells or by purified protein kinase C in vitro. Further, it was observed that PDGF mimicked the action of phorbol diesters to inhibit the EGF-dependent tyrosine phosphorylation of the EGF receptor in [32P]phosphate-labeled fibroblasts. These results are consistent with the hypothesis that increases in diacylglycerol and Ca2+ levels caused by addition of PDGF to fibroblasts activate protein kinase C and that this kinase, at least in part, mediates the effect of PDGF on the phosphorylation of the EGF receptor. The data further suggest that protein kinase C may play an important role in the regulation of cellular metabolism and proliferation by PDGF. PMID- 2987963 TI - Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA. AB - We previously found that a form of Epstein-Barr virus with rearranged DNA induces replication of latent Epstein-Barr virus. We now have found that one of three fragments of this rearranged DNA, when cloned in recombinant plasmids and used to transfect cells, can activate expression of several polypeptides from a latent viral genome. The 33-kDa protein that is the product of the active fragment is likely to be responsible for disruption of latency. PMID- 2987964 TI - Two-photon spectroscopy of locked-11-cis-rhodopsin: evidence for a protonated Schiff base in a neutral protein binding site. AB - We studied the nature of the protein binding site of rhodopsin, using two-photon spectroscopy to assign the location of the low-lying "covalent" 1Ag*- -like pi pi * state in a model rhodopsin containing a locked-11-cis chromophore. The two photon thermal lens maximum is observed at 22,800 cm-1, approximately equal to 2000 cm-1 above the one-photon absorption maximum, indicating that the protein environment has induced a level ordering reversal of the low-lying pi pi * states relative to that observed in retinyl Schiff bases in solution. The spectroscopic results clearly indicate that the chromophore is protonated and that the binding site is uncharged. Electrostatic energy contour maps of the binding site are calculated, showing possible locations for the external counterion(s). Two models of the binding site are proposed that accommodate the available spectroscopic data. One model involves a protonated Schiff base chromophore stabilized by a single negatively charged aspartic or glutamic acid residue. A more complicated model involving two residues (one charged, the other neutral) is also proposed. The latter model is interesting because it also accommodates the observed deuterium isotope effect in the form of a proton translocation between the two residues. The translocation is assumed to be a ground state process, initiated subsequent to the photoisomerization of the chromophore and energetically driven via destabilization of the counterion environment as a result of isomerization induced charge separation. PMID- 2987965 TI - Stimulation of in vitro human skin collagenase expression by platelet-derived growth factor. AB - Platelet-derived growth factor (PDGF) is both chemoattractant and mitogenic for stromal cells. Here, we examined the effects of PDGF on collagenase expression by normal human skin fibroblasts. Culturing cells for 24 hr in the presence of PDGF at 0-180 ng/ml resulted in a dose-dependent, saturable increase in collagenase activity in the culture medium that was paralleled by equal increases in immunoreactive collagenase protein, suggesting enhanced synthesis of a catalytically unaltered enzyme. The specificity of this effect was demonstrated by comparing the collagenase-stimulatory effect with that on total protein synthesis and DNA synthesis. Under in vitro conditions that produced a 2.5-fold increase in collagenase synthesis, there was an approximately equal to 20% increase in total protein synthesis and no change in DNA synthesis. In addition, platelet factor 4, another platelet-derived protein, caused a less than 20% increase in collagenase expression. In time-course studies, stimulation of collagenase synthesis was first observed 8-10 hr after exposure to the growth factor. Conversely, when cells were primed with PDGF for approximately equal to 24 hr and the stimulator was then removed, an increased rate of synthesis was seen for an additional approximately equal to 6 hr, after which the rate reverted to control levels. Since the kinetic data suggested a possible pretranslational effect, fibroblasts cultured with PDGF were used to prepare mRNA. In cell-free translation, total protein synthesis was essentially unaltered; however, the growth factor caused a greater than 2-fold increase in translatable collagenase mRNA. The data suggest that PDGF specifically modulates collagenase synthesis, possibly through a series of events that lead to increased transcription or preferential translation of collagenase mRNA. PMID- 2987966 TI - In situ localization of DNA topoisomerase II, a major polypeptide component of the Drosophila nuclear matrix fraction. AB - DNA topoisomerase II has been immunochemically identified on protein blots as a major polypeptide component of the Drosophila nuclear matrix-pore complex-lamina fraction. Indirect immunofluorescence analyses of larval cryosections have confirmed the nuclear localization of topoisomerase II in situ. Although apparently excluded from the nucleolus, the topoisomerase protein is otherwise distributed throughout the interior of interphase nuclei. Similar immunocytochemical studies performed with permeabilized whole giant cells from third-instar larval salivary glands have shown topoisomerase II to be largely restricted to the polytene chromosomes. Upon nuclear disassembly during mitosis, the topoisomerase polypeptide appears to redistribute diffusely throughout the cell. Faint immunofluorescent staining of mitotic chromosomes is also observed. PMID- 2987967 TI - Rates of evolution of the retroviral oncogene of Moloney murine sarcoma virus and of its cellular homologues. AB - A method is proposed for computing the rates of nucleotide substitution for an oncogene of a retrovirus (v-onc), its cellular homologue (c-onc), and the retrovirus genome simultaneously. The method has been applied to DNA sequences of the v-mos gene of Moloney murine sarcoma virus (Mo-MuSV) and the c-mos and gag genes of Mo-MuSV and Moloney murine leukemia virus (Mo-MuLV). The rates of nucleotide substitution for c-mos, the gag gene, and v-mos are estimated to be 1.71 X 10(-9), 6.3 X 10(-4), and 1.31 X 10(-3) per site per year, respectively. The rate of evolution of c-mos is comparable to that of many functional genes in DNA genomes, suggesting some important biological function played by cellular oncogenes. The rates of nucleotide substitution in the v-mos and gag genes are very high and are similar to those of RNA viral genes such as the hemagglutinin and neuraminidase genes in the influenza A virus. Thus, oncogenes seem to exemplify a general feature of genome evolution: the rate of evolution of RNA genomes can be more than a million times greater than that of DNA genomes because of a high mutation rate in the RNA genome. PMID- 2987970 TI - Loss of sensitivity to ACTH of adrenocortical cells isolated from maturing domestic fowl. AB - Maturation of domestic fowl corticosteroidogenesis was evaluated using purified adrenocortical cells. Basal corticosterone production decreased steadily from 2 days to 26 weeks after hatching. However, maximally stimulated corticosterone production was not changed. In contrast, the half-maximal steroidogenic concentrations (ED50 values or effective doses for 50% maximal effect) of ACTH analogs increased approximately 40 times by 26 weeks, but the ED50 values of 8 bromo-cyclic AMP and pregnenolone were not changed. This suggests that adrenocortical cell sensitivity to ACTH decreases with maturation of the domestic fowl. PMID- 2987971 TI - Sites of action of beta-melanocyte stimulating hormone in aldosterone biosynthesis in the rat. AB - The sites of action of beta-melanocyte stimulating hormone (beta-MSH) on aldosterone biosynthesis were studied using collagenase-dispersed adrenal glomerulosa cells from rats maintained on either normal or sodium-deficient diets for 2 weeks. Isolated cells were treated with a cyanoketone derivative (WIN 19,578) to isolate the early and late steps in aldosterone biosynthesis. WIN 19,578 (1 microM) completely blocked aldosterone production stimulated by sodium depletion, AII, ACTH, and beta-MSH. beta-MSH (1 microM) significantly stimulated pregnenolone production (early step) and the conversion of corticosterone to aldosterone (late step) in aldosterone biosynthesis. The effect of beta-MSH was similar to AII and ACTH. Sodium depletion enhanced the effect of beta-MSH only on the late step in aldosterone biosynthesis. In conclusion, beta-MSH stimulates both the early and late steps of aldosterone biosynthesis. These results suggest that beta-MSH or peptides containing beta-MSH may play a role in the regulation of aldosterone production. PMID- 2987969 TI - Potentiation of antiherpetic activity of acyclovir by ribonucleotide reductase inhibition. AB - Compound A723U, a 2-acetylpyridine thiosemicarbazone, produced apparent inactivation of herpes simplex virus type 1 (HSV-1) ribonucleotide reductase. Inactivation occurred after A723U formed a reversible complex with the enzyme and only while the enzyme was catalyzing the formation of deoxynucleotides. A723U inhibited HSV-1 replication at concentrations that were not toxic to the confluent host cells. Most importantly, A723U and acyclovir (ACV) were found to exhibit mutual potentiation of their antiviral activities. Subinhibitory concentrations of either compound greatly reduced the ED50 (median effective dose) of the other. Studies of the deoxynucleotide pool sizes and the levels of ACV triphosphate (ACV-P3) revealed that A723U not only significantly reduced the pool of dGTP but also increased the level of ACV-P3 in infected cells. The net result was an 80-fold increase in the ratio of ACV-P3 to dGTP. This should greatly facilitate the initial binding of ACV-P3 to HSV-1 DNA polymerase and probably accounts for the mechanism of potentiation. PMID- 2987968 TI - Interleukin 2 (IL-2) augments transcription of the IL-2 receptor gene. AB - We demonstrate that purified interleukin 2 (IL-2) can directly upregulate IL-2 receptor expression on phytohemagglutinin-activated T lymphocytes maintained in culture until IL-2 receptor expression had markedly declined. The IL-2-induced increase in IL-2 receptor number is maximal within 12 hr, requires new RNA and protein synthesis, and is mediated by an interaction of ligand with the high affinity receptors for IL-2. IL-2 stimulation results in increased accumulation of IL-2 receptor mRNA within 4 hr, while an increase in IL-2 receptor gene transcription is detected within 30 min in isolated nuclei. In addition, IL-2 incubation results in increased amounts of c-myc and transferrin receptor mRNA, but it does not augment levels of mRNA encoding the beta chain of the T-cell receptor for antigen. These results demonstrate that IL-2 can directly upregulate transcription and expression of its own receptor and, therefore, indicate that IL 2 may regulate IL-2-dependent immune responses, in part, by influencing the expression of IL-2 receptors. PMID- 2987973 TI - Hemadsorption focus assay for growth of influenza and parainfluenza viruses in human dermal fibroblasts. AB - An assay for virus-induced hemadsorption foci in cultures of human dermal fibroblasts is described. The assay allows determination of activity of species specific as well as nonspecific antiviral agents or biological factors, such as interferons. The assay may be used for abortive (e.g., influenza virus) and productive (e.g., parainfluenza virus) infections of human fibroblasts. PMID- 2987972 TI - Changes in Epstein-Barr virus antibody titers associated with aging. AB - Antibody titers to the Epstein-Barr virus (EBV), early antigen (EA) IgG, and virus capsid antigen (VCA) IgG and IgA, were measured in 44 geriatric subjects to determine if the depression in cellular immunity known to be associated with aging affects the expression of latent EBV. Similar assays were performed on plasma obtained from a young adult (medical student) population as a control group. We found that 89% of the geriatric samples were positive for EA IgG, and 83% of the plasma obtained from medical students were positive for EA IgG. One hundred percent of the geriatric subjects were positive for VCA IgG, and 87% of the medical students were positive for VCA IgG. Seven percent of the medical student blood samples were positive for VCA IgA; in contrast, 36% of the blood samples obtained from the geriatrics subjects were positive. Significant differences were also found in the geometric mean titers (GMT) of antibodies to EBV antigens; the GMT to EBV EA and VCA were significantly higher in the geriatric group. The data suggest that there may be some loss of control over latent EBV by the cellular immune response in geriatric individuals. PMID- 2987974 TI - Morphophysiological studies of the mammalian vestibular labyrinth. PMID- 2987975 TI - An electrophysiological investigation of the rat medial vestibular nucleus in vitro. PMID- 2987976 TI - Synaptic integration in a single neuron. PMID- 2987977 TI - Transformations of exogenous arachidonic acid in human platelets in the presence of oleic- and eicosapentaenoic acids. AB - Effects of various concentrations (12.5-50 microM) of eicosapentaenoic acid (EPA) on the metabolism of exogenous arachidonic acid (AA) in washed human platelets were examined under two different incubation conditions. First platelets were pretreated with EPA before exposing them to labelled AA. In the second incubation platelets were exposed to a mixture of EPA (12.5 and 25 microM) and labelled AA. At all concentrations EPA reduced thromboxane B2 (TxB2) formation, and at 25 and 50 microM this fatty acid reduced also prostaglandins (PGE2, PGF2 alpha). A reduced formation of PGs and TxB2 was confirmed by a decreased formation of PG endoperoxides. While in EPA treated platelets less CO-(TxB2, HHT, PGs) and lipoxygenase (HETE) products were formed at 25 and 50 microM, more HETE seemed to be generated at lower EPA concentration (12.5 microM). Oleic acid reduced TxB2 formation at high concentration (100 microM). EPA showed a dose-dependent inhibition of platelet aggregation induced by arachidonate, epinephrine and collagen; it was most effective against AA-induced aggregation. PMID- 2987978 TI - Potentiating effects of leukotriene B4 and prostaglandin E2 on urinary sodium excretion by the dog kidney. AB - A synergistic vasodilatation was recently demonstrated with leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) in the cutaneous microcirculation. The present study addresses this question to the renal microcirculation, with respect to its eventual influence on the net transport of sodium, infusing small doses of LTB4 (100 ng/min) and PGE2 (3 ng/min) in the left renal artery, and using the contralateral kidney as control in anesthetized Mongrel dogs. In group 1 (hydropenic animals), LTB4 alone failed to influence natriuresis (UNaV) while PGE2 increased UNaV from 152 +/- 20 to 225 +/- 18 uEq/min. The combined infusion of LTB4 and PGE2 resulted in a marked elevation of natriuresis to 368 +/- 26, 317 +/- 30 and 342 +/- 52 uEq/min. In group 2, water diuresis was induced to examine the eventual site of action of these compounds, and to assess the response of the diluting segment of the nephron. In these dogs, PGE2 was first administered and UNaV rose modestly from 51 +/- 12 to 77 +/- 15 uEq/min. LTB4 again had no significant influence on UNaV, but LTB4 and PGE2 produced a marked increment from 75 +/- 16 to 374 +/- 24 uEq/min. Urine volume, as well as free water clearance, increased from 3.3 +/- 0.2 to 6.5 +/- 0.7 ml/min, and from 2.3 +/- 0.2 to 3.7 +/- 0.4 ml/min, respectively, during LTB4 + PGE2. No significant change occurred in the right control kidney during these manoeuvers. Since renal hemodynamics (glomerular filtration and plasma flow) remained relatively stable in both groups of dogs, it is suggested that the combined infusion of LTB4 and PGE2 exerts a direct influence on the net transport of sodium, probably in the proximal tubule, as inferred by the results obtained in group 2. These two important metabolites of arachidonic acid could be involved in the modulation of renal sodium excretion under normal and/or pathophysiological conditions. PMID- 2987979 TI - A metabolite of riboflavin binds to the 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) receptor. AB - A cytosolic receptor to which TCDD binds with high affinity has been reported in a number of rodent tissues, but the endogenous/natural ligand is not known. However, lumichrome, a metabolite of riboflavin, competed with 3H-TCDD for binding to the cytosolic receptor. Neither riboflavin nor riboflavin-5-phosphate competed for binding to the TCDD receptor in rat liver cytosol. Lumichrome is the first known endogenous chemical in the rat to exhibit affinity for the TCDD receptor. PMID- 2987980 TI - Control of 2,3,7,8-tetrachlorodibenzo-p-dioxin binding protein(s) in the hamster kidney. AB - Hamster renal cytosol binds [3H]-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with high specificity. Sucrose density gradient centrifugation revealed two binding entities- one with a low sedimentation coefficient of 4-5 S which was displaced by neither TCDD nor other polycyclic aromatic hydrocarbons (PAHs) and another with a high sedimentation coefficient of 7-8 S which was displaced by TCDD, benzo[a]pyrene (BP), 2-methylcholanthrene (MC), and 7,12 dimethylbenzo[a]anthracene (DMBA) but not by estradiol (E), progesterone (P), cortisol (F), testosterone (T), 5 alpha-dihydrotestosterone (DHT), or methyltrienolone (R-1881), a synthetic androgen. Cytosol from intact male hamsters showed maximum binding of labelled TCDD to the 7-8 S binding site. Castration or hypophysectomy reduced this binding. Pretreatment with DMBA increased binding, whereas diethylstilbestrol (DES) decreased binding. Sex difference was observed in the binding capacity of renal cytosol. This is the first report of endocrine control over TCDD binding and its modulation by other PAHs and steroids. PMID- 2987981 TI - Kynurenine may directly interact with GABA receptors in rat brain. AB - Previous studies have shown that kynurenine may have convulsant activity. In the present investigation the intracerebroventricular injection of L- but not D kynurenine induced convulsions in the rat. In vitro, L- but not D-kynurenine was able to displace 3H-GABA from rat brain membrane preparations. The action was specific for 3H-GABA and was not observed with other ligands. The data suggest that the convulsant activity of L-kynurenine might be due to an interaction with GABA receptors. PMID- 2987982 TI - Motor responses of cyproterone acetate-treated rat vas deferens to electrical field stimulation, noradrenaline and barium chloride. AB - The effect of cyproterone acetate (CA) was studied on the autonomic transmission and smooth muscle function of the rat vas deferens. In the CA-treated vas deferens, the endogenously released noradrenaline (NA) in response to electrical field stimulation or exogenously administered NA produced rhythmic contractions (lacking a tonic component) which were abolished by prazosin - a preferential postjunctional alpha-adrenoceptor blocker. The biphasic nature of the nonadrenergic response elicited on electrical field stimulation in CA-treated rats remained unaltered. Like in controls, yohimbine potentiated this response thereby demonstrating the presence of functional presynaptic alpha-adrenoceptors (inhibitory in nature). The postjunctional alteration on the membrane function appears to be specific to adrenergic stimulation alone, since both tonic and phasic components were demonstrated in the responses resulting from nonadrenergic stimulation or those produced by barium chloride. PMID- 2987983 TI - Cellular transformation by human T cell leukemia retroviruses (HTLV). PMID- 2987984 TI - Cellular transformation by avian viruses. PMID- 2987985 TI - Photoinactivation of a Chlamydomonas mutant (NL-11) in the presence of methionine: roles of H2O2 and O2-. PMID- 2987986 TI - Opioid modulation of ingestive behaviors in woodchucks and racoons. AB - We have examined the effect on feeding of opioid blockade with naloxone in two species which demonstrate a marked seasonality in their feeding patterns, the racoon (Procyon lotor) and the woodchuck (Marmota monax). Naloxone suppressed food intake in the woodchuck which is a true hibernator. Naloxone failed to suppress food intake in the racoon and, in fact, enhanced intake of a preferred sucrose solution. In the racoon, ir-dynorphine concentrations were extremely high in the hypothalamus compared to the values obtained in rats and woodchucks. We suggest that possible explanations for the lack of responsiveness to opiates in racoons may be their extremely high daily food intake relatively to body mass when compared to woodchucks and rats and the high levels of ir-dynorphin may be sufficient to overcome the inhibitory effect of naloxone. These studies stress the occurrence of species diversity in the response to opioid antagonism. PMID- 2987987 TI - Conditioned taste aversion suppresses induction of delayed-type hypersensitivity immune reactions. AB - Conditioned taste aversion was induced in mice by pairing saccharin drinking with an intraperitoneal injection of lithium chloride, a toxic but nonimmunosuppressive drug. Conditioned mice showed not only suppressed saccharin drinking but also a 75% reduction in the induction of delayed-type hypersensitivity immune responses to low doses of sheep erythrocytes. This effect was observed with doses of lithium chloride which had no effect of their own on immune functions. In addition, a reduction in water consumption was not responsible for the reduced immune response of conditioned mice since the immune responses of water deprived mice did not differ from those of nondeprived mice. Conditioned mice exposed to saccharin had higher plasma levels of glucocorticoids than nonconditioned mice, suggesting that the experience of being reexposed to a taste paired with lithium chloride was perceived as aversive. These data demonstrate that alterations in immune functions can be induced by a conditioned taste aversion procedure independently of any immunosuppressive drug. PMID- 2987988 TI - Olfactory cues and pig agonistic behavior: evidence for a submissive pheromone. AB - One hundred and two prepubertal pigs were used in two experiments to determine if adrenocorticotropin hormone (ACTH)-induced increase in submissive behavior could be mediated by odorous signals. In experiment one, urine was collected from pigs treated with either 0, 1 or 10 IU/kg ACTH. Urine from pigs given 1.0 IU/kg ACTH caused a trend for a rise in submissive behavior. Level of plasma cortisol from donor pigs correlated well (r = .92) with duration of submissive behavior in the tested pigs. In experiment two, urine from ACTH-treated pigs increased submissive behavior when sprayed in the air during late fight. Thus, ACTH-induced submissiveness may be mediated by a pheromone. These results fit the hypothesis that, in addition to visual cues, an olfactory cue (perhaps adrenal in origin) is released towards the end of a fight to signal submission. Aerosolizing urine from ACTH-treated pigs may have interfered with this pheromonal signal. PMID- 2987989 TI - Responses of olfactory bulb neurons to repeated odor stimulations in awake freely breathing rabbits. AB - Thirty-one olfactory bulb neurons were recorded in the olfactory bulbs of unanaesthetized rabbits during repeated stimulations. Their single-unit activity associated with the inspiratory phases of the respiratory cycles and that associated with the expiratory phases were processed separately. When responses were classified into 3 types, i.e., excitation, inhibition and null, it was found that a large number of neurons presented variable responses to repeated stimulations with the same stimulus. However, the passage from one type to another was found to be limited: responses by excitation or inhibition to the first stimulation turned into null responses only; null responses turned into either excitation or inhibition. Inspiration- and expiration-related responses were also subjected to a principal component analysis in order to determine whether changes in responses were compatible with a reliable coding of the qualitative properties of a stimulus. The results indicated that the repeated presentations of an odorant induced fairly similar profiles of activity across the set of neurons while different odorants induced clearly discriminable profiles. It is concluded that repeated stimulations do not blur the characteristic features of the across-neuron profile of response of an odorant in the olfactory bulb despite the variability of the responses of the neurons which compose the profile. PMID- 2987990 TI - Multi-dimensional analyses of behavior in mice treated with naltrexone. AB - The effects of naltrexone on the behavior in mice were investigated by using a multi-dimensional behavioral analyser. Within 15 min following observation, naltrexone preferentially suppressed the linear locomotion at the 10 and 30 mg/kg doses. The results suggest that naltrexone selectively disrupts the linear locomotion without affecting other behaviors in mice. PMID- 2987991 TI - Tn5 insertions in the agrocin 84 plasmid: the conjugal nature of pAgK84 and the locations of determinants for transfer and agrocin 84 production. AB - The kanamycin-resistance transposon Tn5 was randomly introduced into pAgK84, a 47.7-kb plasmid coding for agrocin 84 production in Agrobacterium. Using such marked plasmids, pAgK84 was found to be conjugal. It could be transferred to several Agrobacterium strains including those harboring octopine- or nopaline type Ti plasmids. Its presence has no effect on Ti plasmid functions such as opine utilization and tumorigenicity, but it does confer agrocin 84 immunity upon previously sensitive strains. The plasmid could also be conjugally transferred to a Nod+ Fix+ strain of Rhizobium meliloti. The production of agrocin 84 is expressed in all Agrobacterium and Rhizobium transconjugants tested. The agrocin plasmid could not be introduced into restrictionless Escherichia coli or Pseudomonas aeruginosa recipients by conjugation or transformation. The sites of 92 independent Tn5 insertions were mapped on pAgK84. These insertions are dispersed over the entire length of the plasmid. Analysis of the sites and effects of the Tn5 insertions has allowed us to construct a functional map of pAgK84. Forty-three of these insertions, spanning a 20-kb segment of the plasmid, abolished or greatly reduced the production of agrocin 84. The presence of two insertions within this segment having an effect on agrocin production suggests that at least three regions of the plasmid are involved in agrocin 84 biosynthesis. Fourteen of the Tn5 insertion derivatives are no longer conjugally transferable. These insertions all map to a single region of the plasmid and define about 3.5-kb as being associated with transfer functions. PMID- 2987992 TI - Rhizobium meliloti carries two megaplasmids. AB - In Rhizobium meliloti strain 41 the existence of a second megaplasmid (pRme41c) with a molecular weight similar to the sym megaplasmid pRme41b was demonstrated. Derivatives of the wild-type strain carrying pRme41b or pRme41c tagged with Tn5 allowed the examination of the transfer ability of both megaplasmids. The introduction of megaplasmids into the wild-type R. meliloti was not detected, probably because of the action of plasmid genes coding for entry exclusion of the same type of plasmid. However, transmissibility of both megaplasmids was observed in matings with Nod- or Fix- pRme41b deletion mutant recipients and with Agrobacterium tumefaciens at frequencies of 10(-6) - 10(-8). Introduction of the megaplasmids into the R. meliloti recipients resulted in the loss of the same plasmid. On the other hand, pRme41b and pRme41c were compatible. From the extent of deletions in various Nod- and Fix- mutants a DNA region carrying genes probably involved in "surface exclusion" on pRme41b was located. This DNA region is about 50 kb distant from the nod genes and exhibits strong homology with a DNA segment of pRme41c. Symbiotic genes on pRme41c were not identified. PMID- 2987993 TI - Location of rep and inc sequences in the F secondary replicon. AB - Miniplasmids derived by deletion of DNA from the F plasmid secondary replicon have been tested for the ability to replicate and to express incompatibility with the IncFI plasmid, ColV3-K30. The results demonstrate that the minimal rep region of the secondary replicon lies within a 1.9-kb sequence (33.7F-35.6F kb), and that an inc region, presumably involved in replication control, is present in a 0.45-kb portion of the rep region (33.7F-34.15F kb). In addition, the secondary replicon was found not to require DNA polymerase I activity. PMID- 2987994 TI - Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. AB - Derivatives of plasmid pRK290 that are useful for cloning and for analyzing gene expression in a wide variety of Gram-negative bacteria are described. A smaller broad host range plasmid derived from RK2, with properties similar to that of pRK290, is also described. PMID- 2987995 TI - Identification of an Rts1 DNA fragment conferring temperature-dependent instability to vector plasmids. AB - The multiphenotypic drug resistance factor Rts1 expresses a temperature-dependent instability characteristic. This plasmid was digested with the restriction enzyme BamHI. A DNA fragment with a molecular weight of 5.6 MDa (the H fragment) was inserted into plasmid pBR322 (pFK896) or into pSC105 (pYH156) at the BamHI site. These plasmids were unstable at 42 degrees C but stable at 32 degrees C. A restriction-enzyme map of the H fragment was constructed and the instability phenotype (Tdi) was localized to a DNA fragment with 0.5 MDa molecular weight. The temperature-dependent loss of the unstable plasmid pFK896 is abrupt and no gradual plasmid loss of this multicopy recombinant plasmid is observed. The possibility that the Tdi phenotype is due to overgrowth of R- cells was eliminated. PMID- 2987996 TI - R1767, an example of the evolution of resistance plasmids. AB - The Salmonella R-factor system R1767 undergoes frequent rearrangement of its plasmid components. The flux of genetic material within this plasmid system depends on a combination of illegitimate and homologous recombination. The presence of several copies of IS160 and two multiresistance transposons, Tn2410 and Tn2411, are substantial reasons for the observed variations. PMID- 2987998 TI - A vehicle for the introduction of transposons into plant-associated pseudomonads. AB - A recombinant plasmid with wide-host-range transfer functions, narrow-host-range replication functions, and carrying a kanamycin-resistant transposon transferred kanamycin resistance to a number of plant-associated pseudomonads. Southern hybridization studies suggest that only a small portion of the plasmid, coinciding with the location of the transposon, is present in the kanamycin resistant Pseudomonas derivatives. The plasmid sequences appear to be inserted at a number of different sites in the recipient genome. This plasmid can thus be used as a vehicle for the introduction of transposons into some plant-associated pseudomonads and should be useful in both genetic and ecological studies of these bacteria. PMID- 2987997 TI - Comparison of the transposon-like structures encoding clindamycin resistance in Bacteroides R-plasmids. AB - The R-plasmids pBF4, pBFTM10, and pBI136 encode transmissible clindamycin resistance (Ccr) in Bacteroides spp. These plasmids are distinct replicons but the regions implicated in Ccr share some homology and appear to have a transposon like structure. To better understand the mechanism of dissemination and to locate the Ccr determinant(s), the genetic and structural properties of the Ccr regions of each plasmid were compared and contrasted. For this work a single EcoRI restriction fragment containing the Ccr region from each plasmid was cloned into pBR322 in Escherichia coli. Results of restriction mapping and heteroduplex experiments showed that the pBF4 EcoRI-D and pBFTM10 EcoRI-B fragments shared more than 90% base sequence homology but that the EcoRI-C fragment of pBI136 had diverged significantly. The pBI136 fragment also did not confer tetracycline resistance in E. coli as shown for the pBF4 EcoRI-D fragment (D.G. Guiney, P. Hasegawa, and C. E. Davis, 1984, Plasmid 11, 248-252). Heteroduplex experiments showed that the pBI136 EcoRI-C and pBF4 EcoRI-D fragments shared a 1.2-kb region of homology attributed to a directly repeated sequence which bounds the Ccr region. Southern hybridization studies indicated that an additional 0.85 kb of the pBI136 EcoRI-C fragment was homologous to the EcoRI-D fragment of pBF4. This region was characterized by its sequential restriction endonuclease sites for HindIII, AvaII, and DdeI, and it is proposed that the Ccr gene(s) resides in this area. PMID- 2988000 TI - A physical and genetic map of the IncFI plasmid ColV2-K94. AB - A restriction enzyme map of the IncFI plasmid ColV2-K94 was generated using EcoRI, BamHI, HindIII, and XhoI; the genetic features of this element were then mapped from previous heteroduplex studies. PMID- 2987999 TI - Relationships of the Col plasmids E2, E3, E4, E5, E6, and E7: restriction mapping and colicin gene fusions. AB - Thirteen ColE plasmids representing the E2-E7 types have been compared by restriction mapping. Over 80% of their restriction sites were found to be similarly positioned, indicating that these plasmids share a common structure. Three variants are ColE2-CA42 and ColE7-K317, both of which contain 1.8-kb DNA segments in place of a 2.5-kb segment common to the other plasmids, and ColE6 CT14, which has an additional 5.0-kb DNA segment compared to the other plasmids. The colicin (col), immunity (imm), and colicin release (hic) genes of these plasmids have been localized to regions corresponding to those known for ColE3 CA38 and ColE2-P9, with the imm and hic genes adjacent to the 3' end of the col gene. Active colicin is produced from hybrid col genes containing 5' and 3' ends from different E-type plasmids. The 3'-termini of the fused col genes specify the colicin type. PMID- 2988001 TI - [Neurophysiologic findings in chronic alcohol abuse]. AB - Neurophysiological examinations of 100 long term alcohol dependent patients who were having neuropsychiatric treatment, showed symptoms of polytopic damage of the peripheral and central nervous system. The results show that for recognition of the damage an extensive diagnostic programme must be used. PMID- 2988003 TI - Modulation of spinal synaptic transmission by beta-melanocyte stimulating hormone (beta-MSH). AB - Synaptic modulation refers to altered excitability of a synapse by a substance that does not produce a spike potential at the synapse. Available evidence points to the conclusion that beta-melanocyte stimulating hormone (beta-MSH) modulates synaptic transmission through monosynaptic pathways in the cat spinal cord. Earlier evidence is reviewed, and new data are presented. In the first experiments populations of cells contributing to a knee jerk were studied using the Lloyd preparation, and MSH was found to increase the monosynaptic reflex. With intracellular single unit recording techniques, beta-MSH was found to facilitate recovery from synaptic transmission. With extracellular single unit recording techniques and iontophoretic methods for drug application, beta-MSH has been found to increase the probability of generation of single spike potentials by alpha-motoneurons in response to orthodromic stimulation. Administration of beta-MSH did not cause spontaneous discharge of alpha-motoneurons. The physiological and pharmacological importance of synaptic modulation is discussed. PMID- 2988002 TI - 3H-Imipramine binding sites in platelets of hospitalized psychiatric patients. AB - The density of platelet 3H-imipramine binding sites has been proposed as a biological marker in psychiatry. We report the range of platelet 3H-imipramine binding in 55 psychiatric patients and 11 control subjects. All blood samples were withdrawn at 2300 h (on the day of hospital admission for patients). With the use of a slight modification of a previously described 3H-imipramine binding method, a mean B max of 1,510 fmole/mg protein (range: 390-5,560; median: 1,450) and a mean Kd of 2.0 nM (range: 0.6-17.0; median: 1.4) were determined for psychiatric patients. For the controls, a mean B max of 1,590 fmole/mg protein (range: 870-2,570; median: 1,440) and a mean Kd of 1.4 nM (range 0.8-2.4; median 1.4) were determined. When patients were subdivided based on ICD-9 psychiatric diagnoses, no significant differences between distinct subgroups of psychiatric patients with respect to B max or Kd values for platelet 3H-imipramine binding could be established. Similarly, no significant difference between psychiatric patients and controls was obtained. PMID- 2988004 TI - The relationship of endogenous ACTH levels to visual-attentional functioning in patients with congenital adrenal hyperplasia. AB - The within subject experimental approach of either doubling cortisone medication dose or withdrawing steroid treatment for 36 hr preceding behavioral testing was found to induce corresponding significantly elevated or suppressed plasma ACTH levels, as measured by radioimmunoassay, in six of eight adults diagnosed as having congenital adrenal hyperplasia (CAH). During the session characterized by elevated ACTH levels, the CAH patients exhibited significantly reduced median reaction times on the Sternberg Item Recognition Task. Their response pattern was suggestive of facilitated visual attentional functioning and/or overt motor response capacity rather than alteration of simple cognitive processing. Moreover, Sternberg performance was significantly correlated with endogenous ACTH levels but not with levels of plasma cortisol or cortisone replacement medication. This enhancement of performance paralleled a previous finding of improved performance on the Sternberg paradigm by normal adults following exogenous administration of ACTH 4-10 (Ward et al., 1979). Further analysis of the Sternberg performance suggests that other variables such as mineralocorticoid treatment, type of CAH impairment, and sex may act to moderate the degree of ACTH related facilitation on this task. Performance on other visual and verbal attention and memory tasks, found earlier to be sensitive indices of exogenous administration of ACTH 4-10 and related fragments, was not significantly altered by manipulation of endogenous ACTH levels in these CAH patients. PMID- 2988005 TI - [Alveolar process atrophy--therapeutic possibilities and perspectives]. PMID- 2988006 TI - [Effect of x-irradiation on the generation of O2- in rat liver microsomes]. AB - A study was made of generation of O2- in NADPH-dependent chain of oxidation of liver microsomes in irradiated rats (7 and 10 Gy). The rate of O2- generation sharply increased at early times after irradiation. The data obtained prompt an assumption that the increase in the rate of O2- generation is perhaps connected with the changes in functioning of both NADPH-cytochrome P-450-reductase and cytochrome P-450. PMID- 2988007 TI - [Sonographic and computed tomographic diagnosis of liver metastases. A review]. AB - This article reviews the diagnosis of liver metastases by sonography and computed tomography. The ultrasonographic and computer tomographic features of hepatic metastases are described emphasizing the various signs of malignancy and the differential diagnosis of focal liver disease. The advantages and disadvantages of each imaging technique as well as a diagnostic approach in the evaluation of hepatic tumors are discussed. PMID- 2988009 TI - [Arterial DSA of liver masses]. AB - A total of 58 patients underwent intra-arterial hepatic DSA. 42 patients with hepatic masses were preselected by ultrasound or CT. In addition a conventional arteriography was performed in 34 of 58 cases. DSA without diagnostic quality occurred in 4 cases, in 4 other cases a diffuse metastatic disease of the liver was not detected in spite of good image quality. Comparing DSA with conventional angiograms our results demonstrated that DSA had a reduced imaging of intrahepatic small vessels but without negative influence on diagnostic value. Intra-hepatic DSA imaging could be improved using on-line image documentation, small size of image-intensifier and contrast material with an iodine concentration of 300 mg/ml. Advantages of DSA were observed in vascularized tumors and indirect splenoportograms. Contrast material with an iodine concentration of 200 mg/ml was sufficient for hepatic "vascular mapping" or demonstration of vascular diseases including main arteries. PMID- 2988008 TI - [Skeletal manifestations of malignant, nonosseous tumors in childhood]. AB - Skeletal involvement in childhood nonosseous tumors can be due to primary involvement, arrosion or metastasis, secondary due to therapy induced alterations or osteomyelitis following diminished immunity. The occurence of bone changes differs widely from those in adults. Neuroblastoma, rhabdomyosarcoma and malignant lymphoma are discussed in detail. Rare tumors are listed for synopsis. As diagnostic screening method skeletal scintigraphy is recommended, whereas in localized disease X-rays should be performed. Beside roentgenmorphology particularly in primary disease-localisation, frequency and age dependency may give essential diagnostic hints. Prognosis depends on primary tumor. PMID- 2988010 TI - Cytomegalovirus colitis in AIDS: radiographic findings in 11 patients. AB - Radiographic findings in 11 proved cases of cytomegalovirus (CMV) colitis were reviewed and correlated with colonoscopic and pathologic findings. Patients were chronically ill homosexual men with multiple systemic opportunistic infections. Endoscopies showed focal or diffuse inflammation, hemorrhagic plaques, and superficial colonic ulcers. Biopsies revealed inflammatory cells associated with cytoplasmic and intranuclear inclusion bodies. Barium enema examinations showed mucosal granularity, superficial erosions, thickened folds, and spasticity. Disease was either diffuse (four patients) or segmental (two patients), or it involved the cecum exclusively (three patients). Two patients had normal barium enema studies. Computed tomography scans in two patients demonstrated marked colonic wall thickening and mucosal ulcerations. In the homosexual population with acquired immunodeficiency syndrome, CMV colitis should be part of the differential diagnosis of diffuse colitis, segmental colitis, or typhlitis. PMID- 2988012 TI - Cholangiocarcinoma complicating primary sclerosing cholangitis: cholangiographic appearances. AB - Cholangiograms from 104 patients (and serial cholangiograms in 66 patients) with primary sclerosing cholangitis (PSC) were reviewed. In 13 patients the additional diagnosis of cholangiocarcinoma was made at biopsy or autopsy. Cholangiograms from patients with both PSC and carcinoma were compared with cholangiograms from patients with PSC alone. Marked dilatation of ducts or ductal segments (100% vs. 24%) and the appearance of a polypoid mass (46% vs. 7%) were common findings in the group of patients whose disease was complicated by malignancy. In the malignant group, polypoid masses were larger, measuring 1 cm or greater in diameter. On serial cholangiograms, four of 15 patients with progressive stricture formation and four of five with progressive ductal dilatation proved to have carcinomas. The frequent occurrence of bile duct carcinoma as a complication of PSC in this group of patients indicates that PSC has a strong tendency to undergo malignant degeneration. Cholangiographic findings which suggest malignant degeneration include markedly dilated ducts or ductal segments, presence of a polypoid mass 1 cm or greater in diameter, and progressive stricture formation or ductal dilatation. PMID- 2988011 TI - Thyroid radiation absorbed dose from diagnostic procedures in U.S. population. AB - According to a 1981 survey of thyroid imaging methods in the United States, radionuclide thyroid scans and uptake studies increased 250%-300% between 1966 and 1981, while the U.S. population increased only 17%. Collective absorbed dose decreased from 18 X 10(6) rad (18 X 10(4) Gy) in 1966 to 13.9 X 10(6) rad (13.9 X 10(4) Gy) in 1981. The decrease was due to the use of iodine 123 and technetium 99m pertechnetate rather than iodine 131 (I-131 was used for 100% of scans and uptake studies in 1966 and 10% and 54%, respectively, in 1981) and also to fewer free-standing thyroid uptake studies (150,000 in 1966 and 33,000 in 1981). Even with reduced usage, I-131 still accounted for 93% of the collective absorbed dose in 1981. If I-131 were eliminated from diagnostic procedures, the annual absorbed dose would decrease to 1.4 X 10(6) rad (1.4 X 10(4) Gy). The number of radiation induced cancer cases would also be reduced. PMID- 2988013 TI - Lingual thyroid gland: sonographic appearance. AB - Thyroid dysgenesis is often manifested by a lingual or sublingual location of the thyroid gland. If radionuclide studies are not readily available or are contraindicated, ultrasound (US) may be used to locate the gland. The US appearance of a lingual thyroid gland and the role of US in the evaluation of suspected thyroid dysgenesis are described. PMID- 2988014 TI - Coxsackievirus myocarditis--with special reference to acute and chronic effects. PMID- 2988015 TI - The cardiovascular actions of morphine and the endogenous opioid peptides. PMID- 2988016 TI - [Regulation of transcription termination in E. coli: antitermination]. PMID- 2988017 TI - [Coupling of adenylate cyclase to cell membrane receptors--with special reference to the role of GTP-binding protein]. PMID- 2988018 TI - Characterisation of the prostanoid receptors mediating contraction of guinea-pig isolated trachea. AB - The receptors mediating prostanoid-induced contraction of guinea-pig isolated trachea have been characterised in terms of a recently proposed general classification of prostanoid receptors. Results obtained on the trachea were compared with those obtained on guinea-pig fundus, which contains a sub-type of PGE2-sensitive (EP-) receptor termed the EP1-receptor, and guinea-pig lung strip, which contains a thromboxane-sensitive or TP-receptor. The following agonists were studied, PGE2, PGF2 alpha and the thromboxane-like agonists U-46619 and Wy17186. The antagonists studied were SC-19220 which selectively blocks EP1 receptors, and AH19437 which selectively blocks TP-receptors. On guinea-pig fundus the rank order of agonist potency was PGE2 greater than PGF2 alpha greater than Wy-17186 approximately equal to U-46619, and responses to all agonists were antagonised by SC-19220 but not by AH19437. On guinea-pig lung strip the rank order of potency was U-46619 greater than Wy17186 much greater than PGF2 alpha greater than PGE2 and responses to all agonists tested were blocked by AH19437 but not by SC-19220. On the trachea, the rank order was PGE2 = U-46619 greater than Wy17186 = PGF2 alpha. SC-19220 antagonised responses to PGE2 and PGF2 alpha, but not those to U-46619 or Wy17186. Conversely, AH19437 antagonised responses to U-46619 and Wy17186 but not those to PGE2 or PGF2 alpha. It is concluded that prostanoid-induced contractions of guinea-pig trachea can be mediated by both EP1 and TP-receptors. PMID- 2988019 TI - Evaluation of inhibitors of eicosanoid synthesis in leukocytes: possible pitfall of using the calcium ionophore A23187 to stimulate 5' lipoxygenase. AB - The effect on arachidonate metabolism of two compounds (BW755C and benoxaprofen) which have been reported to inhibit 5' lipoxygenase in leukocytes has been evaluated in human polymorphonuclear leukocytes (PMN) stimulated with the calcium ionophore A23187 and serum-treated zymosan (STZ). The syntheses of leukotriene B4 (LTB4) and thromboxane B2 (TXB2) from endogenous substrate were determined by specific radioimmunoassays as indicators of 5' lipoxygenase and cyclo-oxygenase activity in the PMN respectively. Benoxaprofen inhibited the synthesis of leukotriene B4 by human PMN stimulated with the calcium ionophore A23187, but it was approximately 5 times less potent than BW755C. However, benoxaprofen (IC50 1.6 X 10(-4)M) was approximately 100 times less potent than BW755C (IC50 1.7 X 10(-6)M) at inhibiting leukotriene B4 synthesis induced by serum-treated zymosan. Both drugs inhibited thromboxane synthesis by leukocytes stimulated with A23187 or serum-treated zymosan at similar concentrations (approximately 5 X 10(-6)M). The data obtained using STZ as stimulus are consistent with previous in vivo studies and indicate that benoxaprofen is a relatively selective inhibitor of cyclo-oxygenase. However, this selectivity was far less apparent when A23187 was used as a stimulus to release the eicosanoids which suggests that this inhibition could be via an indirect mechanism and therefore A23187 should be used with caution as a stimulus of 5' lipoxygenase for evaluating inhibitors of eicosanoid synthesis. PMID- 2988020 TI - Lipoxygenase products: leukotrienes C4, D4, A4's breakdown products and 12-HPETE influence platelet aggregation in vivo. AB - Clinical and pathological observations of diseases concomitant which leukotriene release show consistently an involvement of platelet activation. This effect was hitherto believed to be due to tissue trauma. The aim of the present study was to elucidate whether lipoxygenase products such as leukotrienes C4, D4, 12-HPETE and the breakdown products of leukotriene A4 have a direct pro-aggregatory property of their own. Platelet aggregation was induced by intravascular excitation of fluoresceiniso-thiocyanate-dextran in arterioles of the hamster cheek pouch. "Time to aggregate appearance" was assessed prior and after parenteral application of the studied compounds. LTA4's breakdown products were found to have anti-aggregatory properties, whereas LTC4, D4 and 12-HPETE enhanced platelet aggregability. PMID- 2988021 TI - Double bond requirement for the 5-lipoxygenase pathway. AB - We showed previously that polyenoic fatty acids with double bonds at carbon 5,8,11 are good substrates for the 5-lipoxygenase and also can be converted to LTC and dihydroxy acids. In order to determine whether all three double bonds are necessary for the 5-lipoxygenase-leukotriene pathway we studied 5,8,14 eicosatrienoic and 5,11,14-eicosatrienoic acid. C14-labeled fatty acids were incubated with 10,000 X g supernatant of homogenate of rat basophilic leukemia (RBL-1) cells in the presence of Ca++ at 37 degrees C. 5,11,14-Eicosatrienoic acid was not converted by the 5-lipoxygenase pathway and 5,8,14-eicosatrienoic acid was mainly converted to 5-hydroxy-6,8,14-eicosatrienoic acid (5-HETE). This monohydroxy was identified by UV spectrometry (UV max 235 nm) and GC-mass spectrometry. Incubations with whole homogenate analyzed by HPLC and bioassay showed that no detectable LTC, LTD or LTE was formed. These data indicate that fatty acids which have double bonds at carbon 5 and carbon 8 are readily converted to the 5-hydroperoxide. However double bonds at carbon 5,8 and 11 are necessary for LTA biosynthesis. This study therefore extends the characterization of the double bond requirement of the 5-lipoxygenase-leukotriene pathway. The number of double bonds necessary at each step varies and increases with each step in the pathway. PMID- 2988022 TI - Mechanism of the inhibition of platelet aggregation produced by prostaglandin F2 alpha. AB - The inhibition of human platelet aggregation produced by PGF2 alpha is not specific for thromboxane A2 mimetics. Aggregation waves induced by PAF and thrombin are also inhibited by PGF2 alpha (8 microM); ADP is unaffected. These effects are still seen in platelets from aspirin-treated donors and platelets desensitized to thromboxane-like agonists (e.g. 11,9-epoxymethano PGH2). In contrast the thromboxane receptor antagonist EP 045 (up to 20 microM) had no effect on primary aggregation induced by PAF, thrombin and ADP. We have previously shown that EP 045 (IC50 = 0.5 microM), but not PGF2 alpha (28 microM), displaces the specific binding of [3H] 9,11-epoxymethano PGH2 to washed human platelets. PGF2 alpha produces small increases in cAMP levels, and both this effect and the anti-aggregation are diminished by the adenyl cyclase inhibitor SQ 22536. The rise in cAMP induced by PGF2 alpha is inhibited to a greater extent by the presence of ADP than by thrombin, PAF or a thromboxane mimetic. The ability of aggregating agents to inhibit this increase correlates inversely with their sensitivity to inhibition by PGF2 alpha. We suggest that the very weak effect of PGF2 alpha on cyclic AMP production is sufficient to account for its inhibitory activity, and it is unlikely to be a competitive antagonist at the platelet thromboxane receptor as suggested by others. PMID- 2988023 TI - Leukotrienes C4 and D4 in psoriatic skin lesions. AB - Chemoattractant arachidonate lipoxygenase products have been recovered from the skin lesions of psoriasis, and may play a role in eliciting the intra-epidermal neutrophil infiltrate that characterises this disease. In view of evidence for lipoxygenase activity in psoriasis, the characteristic vasodilation in psoriatic lesions, and the vasodilator properties of leukotriene (LT) C4 and D4 in human skin, the presence of these LTs in psoriatic lesions has been investigated. Skin chamber fluid from abraded psoriatic lesions contained significantly greater amounts of immunoreactive material than that from clinically normal skin, as determined by a double antibody radioimmunoassay (RIA) that uses antiserum cross reacting with both LTC4 and LTD4. Purification of lesional chamber fluid and scale extracts by high performance liquid chromatography (HPLC) and RIA of fractions showed immunoreactivity which co-eluted with standard LTC4 and LTD4. These findings suggest that LTC4 and LTD4 may play a role in mediating the vasodilation and increased blood flow that characterise psoriatic skin lesions. PMID- 2988024 TI - Comparative effects of prostaglandin E2 and prostaglandin E3 on water flow and cyclic AMP in the urinary bladder of the frog, Rana pipiens. AB - Prostaglandins (PGs) modulate osmotic water flow in amphibian urinary bladders. Gas chromatographic analysis of prostaglandin precursors in bladders showed that arachidonic acid represented 13.0 +/- 0.6% and eicosapentaenoic acid 4.3 +/- 0.1% of the total fatty acid content. The effects of PGE2 and PGE3 on basal and arginine vasotocin (AVT) stimulated water flow were compared. Control water flow (1.1 +/- 0.2 mg/min) was increased to 4.6 +/- 0.3 mg/min with AVT (10(-6)M) present. PGE2 (10(-6)M) inhibited both basal and AVT stimulated water flow. In contrast, PGE3 (10(-6)M) stimulated basal water flow and further increased AVT stimulated water flow. Basal adenylate cyclase activity (ACA, 59 +/- 0.3 pmol cyclic AMP/mg protein/10 min) was stimulated by the addition of AVT in the absence or presence of exogenous guanosine 5' triphosphate (GTP, 10(-5)M). Both PGE2 and PGE3 stimulated basal ACA in the absence, but not in the presence of GTP. In the absence of exogenous GTP, PGE2 increased AVT stimulated ACA, whereas PGE3 decreased it. Both prostaglandins inhibited AVT stimulation when GTP was added. The effects of PGE2, PGE3 and AVT on tissue cyclic AMP levels in whole urinary bladders were similar to the effects seen on ACA in the absence of exogenous GTP. The contrasting effects of PGE2 and PGE3 on control water flow appear distinct from their similar effects on ACA. However, PGE2 and PGE3 may regulate AVT stimulation through mechanisms involving cyclic AMP. PMID- 2988025 TI - Combined modality treatment for stage III ovarian carcinoma. AB - Thirty-eight Stage III ovarian carcinoma patients were treated with a combined modality protocol consisting of sequential initial surgery with a maximal tumor reduction, CHAD combination chemotherapy, second look reductive surgery and whole abdominal irradiation. Sixteen patients (42%) had minimal residual tumors (less than 2 cm) after initial surgery (Stage IIIA) and 22 (58%) had large residual tumors (greater than 2 cm) (Stage IIIB). The patients received 3-14 courses of CHAD combination chemotherapy, with a response rate (CR + PR) in the evaluable (Stage IIIB) patients of 91%. Twenty-eight patients had a second attempt of cytoreductive operation (10 Stage IIIA patients and 18 Stage IIIB patients). In 10 patients no residual tumor was found. In another 12 patients residual tumor (less than 2 cm) was found and completely resected, whereas in six patients a complete resection of large residual tumors (greater than 2 cm) was not possible. Twenty-one of the patients also completed a course of whole abdominal radiotherapy. Radiation was well-tolerated with the usual expected amounts of nausea, vomiting, diarrhea and transient leukopenia and thrombocytopenia. 11/21 (52%) of the patients relapsed within the first 18 months after completion of radiotherapy. The actuarial relapse-free survival at 36 months from completions of radiotherapy was 44%. The actuarial survival for the whole group from diagnosis was 43% at 3 years (70% for Stage IIIA and 41% for Stage IIIB). The data indicated that this combined modality protocol is both feasible and well tolerated but its curative potential for patients with advanced ovarian carcinoma is as yet unknown. PMID- 2988026 TI - Erythropoietic recovery in human after extended field radiotherapy. Ferrokinetic studies of patients treated by radiotherapy followed by autologous bone marrow transplantation and of patients treated by moderate doses (20 Gy) of radiotherapy. AB - The ability of human irradiated bone marrow to provide suitable environment for migrating stem cells has been studied in four patients. These had received chemotherapy plus doses of 40 Gy to thoracic vertebrae and sternum, followed by autologous bone marrow transplantation. The erythropoietic activity in the non irradiated areas was increased but it was low in irradiated areas at 3.5 weeks and undetectable at 6 months after autotransplantation. This shows that no nidation of migrating stem cells actively contributed to bone marrow regeneration after 40 Gy and suggests the existence of environmental lesions. No extramedullary erythropoiesis and no bone marrow extension were found in these patients. Four patients who had received doses of 20 Gy to extended bone marrow volume were submitted to ferrokinetic studies from one to 18 years after radiotherapy. In non-irradiated areas the erythropoietic activity was within the normal range. In irradiated areas, it was significantly lower than in the non irradiated areas and was intermediate between the erythropoietic activity in the sacrum of six healthy control subjects and in the irradiated areas of the four autografted patients. Bone marrow extension was found in these patients. This study suggests that bone marrow recovery after irradiation might not be an all-or none phenomenon. It would be of interest to better document the dose-effect relationship. PMID- 2988027 TI - [Detection of hepatocellular carcinoma by liver scintigraphy]. PMID- 2988028 TI - [Hypercholesterolemia and hepatocarcinoma]. PMID- 2988029 TI - [Clinical picture of multiple sclerosis in the magnetic resonance tomogram]. AB - We performed magnetic resonance tomography (MRT) on 22 patients with multiple sclerosis (MS) using varying scan sequences. In 21 patients MRT produced abnormal findings demonstrating multiple lesions, giving varying signals. The MRT appearances of MS have the following characteristics: circular configuration and homogeneous structure of individual lesions, multiple foci, with involvement of all parts of the brain, but with preference for the periventricular region and white matter. This constellation of MRT features is characteristic for MS, but is largely independent of the varying clinical symptoms. The appearance of the individual lesions and the distribution pattern of the foci is sufficiently characteristic to support the clinical diagnosis of MS in younger patients. PMID- 2988030 TI - [Nuclear spin tomographic studies in liver diseases]. AB - Various diseases of the liver were studied by nuclear resonance tomography using a supra-conductive magnet, with 0.5 T field strength, and the value of the method for the diagnosis of these conditions has been studied. MR tomography is superior to CT for differentiating between solid and fluid lesions, such as haemangiomas since, by choosing suitable techniques, high contrast can be achieved without enhancement. Most diffuse parenchymal diseases and heterogeneous malignant tumours show little change in relaxation times and therefore cannot be diagnosed with great sensitivity or specificity. Nuclear magnetic tomography cannot replace the sonographic, CT and nuclear medicine techniques used for examining the liver, but is a useful addition for special problems. PMID- 2988032 TI - [Budd-Chiari syndrome in liver echinococcosis]. AB - In two female patients a diagnosis of Budd-Chiari syndrome secondary to hepatic echinococcosis was established by CT. One patient developed acute Budd-Chiari syndrome secondary to E. granulosus lesions of the liver. The second patient presented with a picture of chronic Budd-Chiari syndrome secondary to alveolar echinococcosis. CT findings of Budd-Chiari syndrome included ascites, low density areas in the liver parenchyma, hypertrophy of the caudate lobe, non visualisation of the hepatic veins, occlusion of the retrohepatic inferior vena cava and enlarged retroperitoneal veins. PMID- 2988031 TI - [Differential diagnosis of liver diseases in the nuclear spin tomogram]. AB - In evaluating nuclear magnetic tomography for the diagnosis of liver disease, one must differentiate between circumscribed and diffuse lesions. Nuclear magnetic tomography provides additional information for lesions which are echogenic on ultrasound and can differentiate between metastases, haemangiomas and hamartomas. In diffuse parenchymal disease measurement of relaxation time can differentiate between fatty liver, cirrhosis (alcoholic, primary biliary), haemochromatosis (cirrhotic transformation) and hepatoma. NMR spectroscopy is a method for the future. PMID- 2988033 TI - [Diagnostic value of computed tomography in stomach cancer]. AB - 60 patients with gastric cancer were checked by computed tomography; 57 underwent surgery. Small tumors could not be demonstrated reliably. Double contrast studies and gastroscopy remain the leading diagnostic procedures. In advanced cases computed tomography informs the surgeon about extragastric tumor expansion and the existence and location of metastatic disease. Nearly all carcinomas with a diameter of more than 2 cm in computed tomography proved to be stage T4 (UICC). In selected cases diagnostic laparotomy could be avoided. Because of the lack of therapeutic alternatives the influence of computed tomography on the regimen is limited. Therefore CT-studies are not absolutely necessary in gastric cancer, but can be very helpful in selected cases. PMID- 2988034 TI - [Computed tomographic morphometry of the normal pancreas in adults]. AB - The relationship of the morphology of the normal pancreas, as seen on CT, and age was studied in 200 normal people aged between 19 and 87 years. The A.P. diameter, vertebral body/pancreas ratio, cranio-caudal diameter, contour and structure of the pancreas were analysed. The most marked age-related changed occurred in the vertebral body/pancreas ratio, but all other measurements showed a reduction dependent on age. With increasing age, the contour of the pancreas becomes more lobulated and, at the same time, the structure changes from an homogeneous to an inhomogeneous appearance. These findings correspond with pathological-anatomical observations. PMID- 2988035 TI - [Bile duct endoprostheses. Insertion using a combined endoscopic-radiologic technic]. AB - Percutaneous transhepatic biliary drainage is a proven method for treating obstructive jaundice due to malignancy. However, the rate of complications for primary internal drainage is relatively high. Consequently, various types of biliary endoprostheses which can be introduced transhepatically or endoscopically have been developed. These prostheses also have serious disadvantages: there is a high risk of dislocation or obstruction. The present paper describes the technique of combined endoscopic-radiological introduction of large biliary prostheses and discusses the advantages of this method. PMID- 2988036 TI - Significance of balloon pressure recording during angioplasty. An experimental study. AB - During angioplasty of artificial stenoses, atherosclerotic human cadaver arteries, and normal canine arteries, pressure and volume of the dilatation balloons were continuously recorded. We found that a sudden yield of a lesion corresponded to a sudden drop in the pressure curve and an increase of the balloon volume. Volume monitoring was insensitive, but pressure recording was very precise. Continuous pressure recording, using a non-compliant inflation system, correctly demonstrated small breaks and ruptures of atherosclerotic vessels, changes not seen on fluoroscopy. PMID- 2988037 TI - [Transvenous DSA following surgery of the supra-aortic trunks]. AB - This paper presents our experience using transvenous DSA for control of vascular, prosthetic or bypass patency following operative intervention in the supraaortic branches. The results in 33 cases examined under outpatient conditions demonstrate that DSA via transvenous administration of a contrast material with high iodine concentration is useful to produce images of diagnostic quality (32/33 patients). The pre-condition is a good patient's cooperation and a sufficient cardiac output. Using this minimal invasive method the operative results can be evaluated and vascular complications can be excluded. PMID- 2988038 TI - [Intravenous DSA following kidney transplantation]. AB - Intravenous DSA is a simple angiographic technique in the follow-up of patients with renal transplants. It is of value in the post-operative anuric phase, for investigating renal function which is deteriorating and for demonstrating, or excluding, stenosis of the renal artery at the anastomosis during the development of hypertension. Changes in the peripheral arteries are not usually demonstrable. Reduced opacification of the parenchyma is observed during rejection and if there has been damage to the transplant, but is not seen if the transplant is functioning normally. PMID- 2988039 TI - [Importance of digital subtraction angiography in the control of dialysis shunts]. AB - The methods, indications and results of imaging haemodialysis access devices, using DSA techniques, are presented, based on 34 examinations. The advantages of this technique with respect to radiation exposure, dynamic registration and dose of contrast medium are compared with conventional film-screen techniques. Case reports illustrate the result and clinical value. DSA is recommended as the method of choice for the common indications for imaging haemodialysis access devices. PMID- 2988040 TI - [Localization diagnosis of enlarged parathyroid glands: 201Tl-99mTc-subtraction scintigraphy compared with 5-MHz-sonography]. AB - Fifty patients clinically suspected of having primary hyperparathyroidism were examined by scintigraphy (201Tl-99mTc subtraction) and sonography (5 MHz). In 28 cases, the results of subtraction scintigraphy, and in 26 cases, the results of sonography were checked by operation. In 25 patients there was a solitary parathyroid adenoma and in two, primary hyperplasia. In one patient all four parathyroid glands were normal. Subtraction scintigraphy had a sensitivity of 90% and a specificity of 98%, while sonography had a sensitivity of 82% and a specificity of 95%. The combination of 201Tl-99mTc subtraction scintigraphy as a functional investigation and sonography as a structural imaging method produced a sensitivity of 96% and is therefore superior to all other methods of localisation for normally situated glands. Diagnostic certainty is reduced by the presence of a nodular thyroid. PMID- 2988041 TI - [Dynamic reflux scintimetry for quantifying gastroesophageal reflux in patients with prolonged esophageal transit]. AB - A combination of a radionuclide transit test and a dynamic gastroesophageal scan was evaluated in normal volunteers, in patients with achalasia treated by pneumatic dilatation (n = 34) or Heller myotomy (n = 21). Interpretation of 31 of 57 examinations done with usual scintiscan was not possible because of too high esophageal tracer retention. Only one case could not be interpreted with the modified technique. Gastroesophageal reflux was detected and quantified in this manner in 8 patients, 6 more than with the usual scintiscan. 7 of these 8 patients have had Heller procedure, 1 patient even combined with fundoplasty. PMID- 2988042 TI - [Skeletal manifestations of generalized mastocytosis in the skeletal scintigram compared with roentgen findings]. AB - Bone scans and skeletal x-rays of eight patients with systemic mastocytosis were reviewed. Mast cell infiltration of bone marrow had been proven histologically in every patient. Bone scan and roentgenographic findings are not specific for the disease and do not correlate well in some patients. A generalized increase of uptake was noted in two patients, a generalized decrease of skeletal activity with poor delineation of bony structures was observed in others. A circumscribed increase of activity was observed in some patients, only one patient had a normal bone scan. Roentgenographic examination revealed diffuse sclerosis of trabecular bone in three patients, osteoporosis with collapse of multiple vertebral bodies in three patients, and no abnormalities in two patients. PMID- 2988043 TI - [Sonography in salivary calculi--indications and status]. AB - In 23 patients with sialolithiasis the value of real time sonography was investigated. The calculi were correctly diagnosed in 91% of the patients. 3 of 4 calculi could be localized in the main duct of the salivary gland. Because of the high accuracy and the possibility to detect non opaque stones, sonography should be performed, if on plain film the localisation of the calculi is questionable, before sialography is done. In sonolithotrypsy, sonography can be used to document the extraglandular localisation of intraductal calculi. PMID- 2988044 TI - [Changes in the uterus following treatment of the mother with diethylstilbestrol ("DES-daughter")]. AB - In the fifties and sixties pregnant women with a history of habitual abortions or premature deliveries were treated with DES (diethylstilboestrol). This treatment can lead to certain pathognomonic uterine changes in the daughters, that are recognizable on hysterosalpingographs. Because of the obstetric consequences, it is important to interpret these changes correctly. The findings in 7 patients are described. As most of the DES-daughters are now coming into reproductive age, gynecologists and radiologists will be confronted more frequently with their specific problems. PMID- 2988045 TI - [Obstetric pelvimetry using digital radiography (CT topogram)]. AB - Radiographic examination during pregnancy and pelvimetry before pregnancy are still required for particular indications. The use of digital radiography (lateral topogram on CT) reduces the radiation dose to the foetus and ovaries by a factor of 10 to 100, as compared with conventional pelvimetry, as described by Guthmann. The accuracy of this method is satisfactory, the technique is simple, it is rapid and is easy for the patient, particularly for pregnant women. As long as pelvimetry is not possible without ionising radiation, we consider pelvimetry with digital radiography, when strictly indicated, to be acceptable. In order to determine the transverse diameter of the pelvis, a single additional CT cut through the narrowest part of the pelvis is suggested. PMID- 2988046 TI - [Syringomyelia on MR]. PMID- 2988047 TI - [Roentgen findings in an angiomatous dysplasia (Weber type)]. PMID- 2988048 TI - [Unilateral polyostotic fibrous dysplasia]. PMID- 2988049 TI - Congenital absence of a pedicle in a thoracic vertebra. PMID- 2988050 TI - [Residual angina following mammary artery bypass due to anomalous origin of the thyrocervical trunk]. PMID- 2988051 TI - [A.v. malformation in the region of the vein of Galen in a newborn infant. Detection by sonography and subtraction angiography]. PMID- 2988052 TI - Spontaneous dissection of the cervical part of internal carotid artery. PMID- 2988053 TI - [Tilting of the epiglottis following thoracic sympathectomy]. PMID- 2988054 TI - [A rare complication of Crohn disease following contrast enema of the small bowel]. PMID- 2988055 TI - [Computed tomographic image of a trichobezoar]. PMID- 2988056 TI - The renovertebral index in kidney size estimation using computerised tomography. PMID- 2988057 TI - [Applicability of digital methods in x-ray diagnosis]. AB - The significance of digital examination methods continues to grow. Since the introduction of computer tomography, two additional digital imaging modalities digital subtraction angiography (DSA) and magnetic resonance imaging (MRI)--have been developed. In this paper we present basic techniques of computerized image processing and outline foreseeable future developments. Furthermore, we discuss the possibilities and problems presented by digital picture archiving and communication systems (PACS). PMID- 2988058 TI - [Computed tomography of the heart. Technic for quantitative evaluation of the perfusion of the ischemic myocardial region. Experimental basis]. AB - A CT standard procedure has been developed in six series of experiments, using 62 anaesthetised dogs in order to quantitative relative blood flow through ischaemic heart muscle. Measurements of cardiac dynamics by means of contrast media were less strictly controlled. The use of the standardised procedures with contrast injections is necessary because of the variable perfusion and washout intervals of the contrast in normal and ischaemic myocardium. PMID- 2988059 TI - [Quantitative cardiac computed tomography]. AB - The scope and limitations of quantitative cardiac CT have been evaluated in a series of experimental and clinical studies. The left ventricular muscle mass was estimated by computed tomography in 19 dogs (using volumetric methods, measurements in two axes and planes and reference volume). There was good correlation with anatomical findings. The end-diastolic volume of the left ventricle was estimated in 22 patients with cardiomyopathies; using angiography as a reference, CT led to systematic under-estimation. It is also shown that ECG triggered magnetic resonance tomography results in improved visualisation and may be expected to improve measurements of cardiac morphology. PMID- 2988060 TI - [Determination of left ventricular global ejection fraction using ECG-gated cardiac computed tomography]. AB - The left ventricular ejection fraction was calculated in 100 patients by means of ECG-gated cardio-CT and the results compared with those obtained by laevo cardiography. A moderately good correlation of r = 0.736 was obtained. In general, CT under-estimated the ejection fraction. The normal range of the ejection fraction obtained by CT was 62.7% +/- 12.4%. Its specificity was high at 96%, but sensitivity was low, with 48.6%. PMID- 2988063 TI - [Results of percutaneous transhepatic bile duct drainage]. AB - Significant reduction in serum bilirubin levels was obtained in 90.7% of 75 patients on whom percutaneous transhepatic biliary drainage had been carried out, either immediately before operation or as a prolonged palliative procedure. The technical requirements, complications, mortality rate and length of stay in hospital are less than following palliative surgery. PMID- 2988064 TI - [Combined radio-/chemotherapy of invasive bladder cancer]. AB - In a prospective study, 19 patients with advanced transitional carcinoma of the bladder (stages T.3 and T.4) were treated by combined radiotherapy and chemotherapy. A total of 450 mg cis-platinum or 1.6 mg/kg body weight were given in four treatment periods with three-week intervals. At the same time, the total dose of 60 Gy was given to the tumour over two periods (telecobalt or high-energy photons). There were few systemic or toxic side effects. Recurrence-free survival during the next 36 months was 76%, which compares with a control group treated by irradiation only, who had a survival of 48%. PMID- 2988062 TI - [Contrast media in magnetic resonance tomography. A review. 1. Physicochemical and pharmacological bases of MR contrast media using gadolinium-DTPA as an example]. AB - The principles underlying contrast media for MR tomography are totally different from those for conventional contrast media. Substances suitable for MR tomography are paramagnetic complexes which alter the relaxation times of the tissues. Signal intensity on the MR images are influenced correspondingly. The mode of action of MR contrast media is explained. Gadolinium-DTPA, the first MR contrast medium in clinical use by means of intravenous injection, is described. Its tolerance has been investigated by animal experiments and its effectiveness as an contrast agent has been studied in normal individuals. In the second part of the paper, early results from the use of gadolinium DTPA are given. The additional information which can be obtained by using MR contrast media is discussed. PMID- 2988061 TI - [Computed tomography, magnetic resonance tomography and echocardiography in hypertrophic cardiomyopathy]. AB - The value of echo-cardiography, CT and magnetic resonance tomography was evaluated in 10 patients with hypertrophic cardiomyopathy. Echocardiography made the diagnosis in most cases. The diagnostic criteria depended on functional and morphological changes. CT and magnetic resonance can show thickening in the myocardium, either symmetrical or asymmetrical. Magnetic resonance tomography provides excellent demonstration of the myocardium and accurate delineation of the degree of hypertrophy. It is well suited for measuring cardiac wall thickness. PMID- 2988065 TI - [Computed tomography of apical lung cancer]. AB - CT was performed on 22 patients with carcinoma of the lung apex. CT provided reliable information concerning the local extent of the disease. Infiltration of the thoracic soft tissues was found in 91%, ribs in 87%, vertebral bodies in 61% and of the spinal canal and mediastinum in 22%. Metastases in the hilar and mediastinal lymph nodes were demonstrated in 38%. In eight patients, as a result of the CT, radiotherapy was performed, followed by resection of the tumour, and in 15 patients radiotherapy alone was used. The CT findings, by which operability or inoperability are judged, and the actual therapeutic procedures are discussed. PMID- 2988066 TI - [Computed tomography of the skeletal muscles in neuromuscular diseases]. AB - CT-documentation of skeletal muscular lesions caused by neuromuscular diseases implies an essential contribution to conventional techniques in the macroscopic field. Size, distribution and degree of lesions as well as compensatory mechanisms are proved thereby. We report about the different effects on muscle appearance referring to 106 patients of our own experience in amyotrophic lateral sclerosis, spinal muscular atrophy, poliomyelitis, polyradiculitis, polyneuropathy as well as peripheral traumatic nerve lesions. PMID- 2988067 TI - [Computed tomography of the orbit. I. Trauma-induced changes]. AB - The CT findings in 120 patients following trauma to the orbits, and in 60 patients with iatrogenic abnormalities, have been analysed. CT has been found to be extremely useful in the diagnosis of trauma and postoperative changes by its ability to demonstrate fractures, foreign bodies, haematomas, abscesses, soft tissue displacement or perforation of orbital structures, as well as showing prosthetic material. PMID- 2988069 TI - [Differential diagnosis of masses in the head of the pancreas area]. AB - Masses in the head of the pancreas area may be caused by neoplastic, inflammatory and a variety of other processes originating within the pancreas or in adjacent structures and organs. CT following intravenous bolus injection of the contrast agent and oral staining of the small bowel allows the precise anatomic localization of the lesion and helps to evaluate its dignity and pathology. Nevertheless, frequently the final can be established only at open surgery and by histopathologic examination of the resected specimen. PMID- 2988070 TI - [Program for calculating diagnostic x-ray spectra]. AB - A code written in BASIC is described which calculates x-ray spectra for tube voltages from 20 to 150 kVDC. Data for the spectral distribution, the mean photon energy, kerma in air, mean energy of the kerma distribution together with 1. and 2. half-value layers for aluminum are obtained. Numerical values for the differential energy intensity function and the Thomson-Whiddington constant were taken from Birch et al. (6). For target angles up to 30 degrees agreement between calculated and experimental data is good. For larger angles some discrepancies appear which could be either due to increased filtration by tungsten (surface roughness) or limited applicability of the data used. The program is tailored to be run on a personal computer. PMID- 2988068 TI - [Computed tomography of the orbit. II. Iatrogenic changes]. AB - The typical CT visualisation of silicone implants and prostheses after ophthalmological operations to the eye and orbit are described. Changes following surgery or trauma, such as haemorrhage, inflammation or abscess, are discussed and the difficulties in differential diagnosis are dealt with. PMID- 2988071 TI - [Intralobar pulmonary sequestration]. PMID- 2988072 TI - [Persistent myocardial sinusoids]. PMID- 2988073 TI - [Compression of the left coronary artery by a calcified subaortic left ventricular diverticulum]. PMID- 2988074 TI - [Detection of an intramuscular wood fragment using sonography]. PMID- 2988075 TI - Dural arteriovenous malformation of the posterior fossa. PMID- 2988076 TI - [CT aspects of the most common variant of the popliteal artery entrapment syndrome]. PMID- 2988077 TI - [Radiologic diagnosis of proximal hepatic artery aneurysms]. PMID- 2988078 TI - Gas in the portal venous system. Illustrated by two case reports. PMID- 2988080 TI - [Paravesical Meckel's diverticulum]. PMID- 2988079 TI - [Sonographic diagnosis of a choledochal cyst in a child]. PMID- 2988081 TI - [Parameters derived from ventriculography using radioactive isotopes applied to the study of acute myocardial infarction]. PMID- 2988082 TI - [Beta-adrenergic receptors in the liver of rats. Direct localization by means of a fluorescent beta-blocker]. PMID- 2988083 TI - [Localization of glucagon receptors in intestinal and vascular smooth muscle fibers in the dog. Use of autoradiographic technics]. PMID- 2988084 TI - [Changes in renal function and the intracellular environment in cirrhotic patients]. PMID- 2988085 TI - [Effects of pepstatin A on neutrophils; cross-deactivation with FMLP]. AB - The ability of pepstatin A, a protease inhibitor produced by Streptomyces testaceus, to elicit a number of responses by the human PMN has been studied. In lysozyme and beta-glucuronidase release, pepstatin A 10(-5)M is equivalent to the synthetic oligopeptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) 10(-7)M. In superoxide release, pepstatin A 10(-5)M produces 80% of that originated by FMLP 10(-7). After two minutes of incubation the superoxide release is important, there being no further increase after 10 minutes. Preincubation of the cells with cytochalasin B before stimulation with pepstatin A elicits a noticeable increase in O2- release. In chemotaxis, pepstatin A 10(-6) originates the same cell motility as FMLP 10(-9). Pepstatin A produces a cross deactivation with FMLP which adds further evidence to the hypothesis that both stimuli compete for the same receptor in the PMN. PMID- 2988086 TI - Effect of dopamine and dopamine-receptor blockade on in vitro renin release, tissue renin content and tissue cyclic AMP content in the rat. AB - This study evaluated the in vitro renin release, tissue cyclic AMP content (TcAMPc), and tissue renin content (TRC) changes with time, in response to administration of dopamine (DOP) and of the dopamine-receptor blocking agent pimozide (PIM) to renal cortical slices from sodium deficient (SD) rats. Addition of 10(-3)M DOP to the slice preparation resulted in a gradual stimulation of RR with time, which was significantly different from that seen in control samples after 60 min of incubation. In contrast, TcAMPc of the DOP-treated samples was significantly greater than that of controls after 5 min of incubation. At 60 min, mean TRC of DOP-treated samples was greater than that of controls but not significantly. Two PIM doses (10(-8)M and 10(-6)M, whether added alone or together with 10(-3)M DOP to the cortical slice system, significantly increased RR in each instance while simultaneously depressing TcAMP content markedly below that of unstimulated controls at all incubation times examined. Mean TRC of pimozide-treated samples was also lower than that of controls by 60 min. These in vitro data in the SD rat suggest that: 1) stimulation of renin release by DOP is time-dependent and is mediated by a TcAMP-generating mechanism, and 2) the increase in renin release by PIM administration appears to involve pharmacological inactivation of TcAMP-generating pathways and disruption of membrane permeability, leading to uncontrolled RR. PMID- 2988087 TI - [Endocrine responses to insulin and gestational age in the calf]. AB - The influence of hypoglycemia on pituitary and cortical and medullary adrenal response was studied in Holstein X Friesian male calves by giving an intravenous injection of bovine insulin (0.2 U/kg b.w.). Seventeen (9 treated and 8 control) calves were born spontaneously at term (278 days) and 9 (5 treated and 4 control) were delivered by caesarean section on day 260 of gestation. Insulin injections were given 1, 5, 10 and 30 days after birth. Plasma insulin and ACTH concentrations were measured by radioimmunoassay, cortisol by competitive protein binding assay, catecholamines by radio-enzymology and glucose by colorimetry (glucose oxydase). In calves born spontaneously at term, the insulin injection induced a significant rise in plasma ACTH and cortisol concentrations on day 5 after birth; this rise occurred on days 10 and 30 after birth in calves delivered on day 260 of gestation. The rise in plasma insulin concentration following insulin injection was always more pronounced and sustained in calves delivered before term than in those born spontaneously at term. Following insulin injection, plasma epinephrine concentration increased significantly only on day 30 in calves born spontaneously at term. After treatment, no other change in plasma catecholamine concentrations was observed in any group of calves at any age. These results indicate that the pituitary-adrenal axis responded similarly in 5-day old calves born spontaneously at term and in 10 (or 30)-day old calves delivered by caesarean section 18 days before the expected time of parturition. PMID- 2988088 TI - Precipitating antibodies in experimental visna and natural progressive pneumonia of sheep. AB - Serological responses of Icelandic sheep experimentally infected with visna virus (vv) were contrasted with responses in American Targhee sheep naturally infected with progressive pneumonia virus (PPV). Precipitating antibodies assayed by immunodiffusion were compared with the neutralising and complementing fixing antibody response. In experimental infections with vv, complement fixing and neutralising antibodies appeared early after infection and rose to high levels in all sheep, while precipitating antibodies were detected only at minimal titre. In natural infections with PPV, immune responses were less consistent and precipitating antibodies were detected more frequently than complement fixing or neutralising antibodies against PPV. These results may suggest important biological differences between the lytic fibroblast-tropic virus strains used for experimental infection of Icelandic sheep and the nonlytic macrophage-tropic strains of PPV circulating in nature. Lytic strains evoke a brisk response against the viral glycoprotein with high titre neutralising antibody while nonlytic strains induce a less consistent response to the glycoprotein. PMID- 2988090 TI - Pathogenesis of canine parvovirus-2 in dogs: histopathology and antigen identification in tissues. AB - The pathogenesis of canine parvovirus-2 was studied in orally inoculated conventional dogs using histopathological and peroxidase anti-peroxidase staining techniques. Lymphoid necrosis and depletion of lymphocytes from lymphoid tissues were most notable on days 5 and 6 after exposure. Lymphocyte hyperplasia occurred following day 7. Epithelial cell changes in segments of the small intestine were more severe on days 6 to 9 after exposure in areas associated with Peyer's patches and in the upper segments of the small intestine. The lymphocyte was the primary infected cell. Virus infected cryptal epithelial cells were not detected until 24 hours after the identification of infected cells in lymphoid tissues on day 4 after exposure. The majority of virus infected epithelial cells were found in crypts intimately associated with or adjacent to Peyer's patches in the upper segments of the small intestine. PMID- 2988089 TI - Pathogenesis of canine parvovirus-2 in dogs: haematology, serology and virus recovery. AB - The pathogenesis of canine parvovirus-2 (CPV-2) was studied in orally inoculated conventional dogs using haematological, serological and virological techniques. Virus was first isolated from mesenteric lymph nodes on day 2 after exposure, tonsil on day 3 and small intestine on day 3. Viraemia occurred subsequently and was present in most dogs on days 4 and 5 after exposure. CPV-2 could be isolated from all tissues during viraemia. Relative pyrexia, lymphopenia and neutropenia occurred on days 5, 6 and 7 after exposure, respectively. Virus excretion in faeces began in most dogs on day 4 and continued despite the appearance of neutralising serum antibody. Specific serum antibody, detected in some dogs as early as day 3 and in all dogs by day 7 after exposure, eliminated viraemia and inhibited virus isolation from tissues in cell culture. PMID- 2988091 TI - Detection of antibodies against infectious bursal disease virus: a comparison of three serological methods. AB - Four different oil emulsion infectious bursal disease virus (IBDV) vaccines were inoculated into four-week-old specific pathogen-free chickens. At weekly intervals for five weeks, sera were obtained from the vaccinated birds and from uninoculated control birds and examined for antibodies against IBDV by enzyme linked immunosorbent assay (ELISA), the quantitative agar gel precipitin (QAGP) test and the virus neutralisation (VN) test. There was a highly significant correlation between the mean responses to all tests; the highest correlation (0.818) was between VN and QAGP and the lowest (0.573) between QAGP and ELISA. Generally the ELISA detected positive sera earlier than the VN test which in turn was more sensitive than the QAGP test. The ELISA and QAGP test were less variable, more reproducible and easier to perform than the VN test. PMID- 2988092 TI - Development of a system for genetic and molecular analysis of Streptococcus agalactiae. AB - DNA-DNA hybridisation was used to compare the genetic relation of human and bovine strains of Streptococcus agalactiae. All strains showed significant homology under very stringent hybridisation conditions. The extent of relatedness did not correlate with the serological type. It was demonstrated that the S faecalis transposon Tn916 could be inserted randomly into the S agalactiae chromosome when introduced by conjugation. The ability of Tn916 insertion to cause genetic changes in S agalactiae was confirmed by identification of a mutation in lactose and trehalose fermentation associated with acquisition of the transposon. This system should be useful in genetic analysis of the pathogenicity of S agalactiae. PMID- 2988093 TI - Pathological and virological studies of experimental parvoviral enteritis in calves. AB - Calves held in isolation showed a progressive decline in maternally derived antibody titres to bovine parvovirus but low concentrations of inhibitors resistant to heat and kaolin treatment persisted as the animals matured. These inhibitors had both haemagglutination inhibition and plaque neutralising activity and were considered to be of non-specific origin. Following oral challenge with bovine parvovirus, calves developed mild to moderate diarrhoea, with lymphopenia and viraemia. Sequential virological and immunofluorescent studies showed that the virus initially infected tonsils and intestinal tract, subsequently spreading to systemic lymphoid tissues. Histological and scanning electron microscopic examinations revealed moderate small intestinal villus atrophy and fusion due to crypt damage, together with lymphoid necrosis predominantly associated with the intestinal tract and thymus. Although the disease was not very severe, this may have been because the low parasite burden in the animals reduced mitotic activity in susceptible tissues. PMID- 2988094 TI - Epidemiological studies of parvovirus infections in calves on endemically infected properties. AB - Bovine parvovirus serology and virus excretion were monitored in calves located on three endemically infected North Queensland properties. Maternally derived serum antibody to bovine parvovirus was found to have a half-life of 19 days. On all three properties, calves developed intestinal bovine parvovirus infection with seroconversion soon after weaning. This occurred more promptly where the environment was subject to heavier bovine parvovirus contamination due to management practices. The concurrent presence of moderate levels of residual serum antibody had only minor influence on the onset of the infection. On one beef cattle property, onset of intestinal bovine parvovirus infection was associated with an outbreak of post-weaning diarrhoea. Anthelmintic treatment trials indicated that this syndrome was unrelated to helminth burdens, though coccidiosis appeared responsible for occasional subsequent cases of dysentery. It was considered that bovine parvovirus may have significantly contributed to the development of the diarrhoea syndrome, in conjunction with substantial weaning stresses. PMID- 2988095 TI - Transformation of cultured equine fibroblasts with a bovine papillomavirus. AB - Fetal equine fibroblasts exposed to bovine papillomavirus became transformed by the criteria of morphological alterations and the acquisition of an increased life span, although they failed to grow in soft agar. Papillomavirus genome persisted in the transformed fibroblasts and was apparently not integrated with the cellular genome. These findings support the notion that bovine papillomaviruses are involved in the production of equine sarcoids. PMID- 2988096 TI - [Subungual glomus tumor. Apropos of a case]. AB - Glomus tumors are rare, though not exceptional. They are considered by some authors as hyperplasia of a normal neuromyo-arterial glomus, by others as a benign tumoral proliferation. Most of them combine the three histological types initially described by Masson. i.e. solid, angiomatous and muco-hyaline. Subungual glomus tumors are more often seen in middle-aged subjects than in children. Although often belatedly diagnosed, they have characteristic symptoms, notably paroxysmal, acute pain provoked by cold or by minimal traumas: the Raynaud's phenomenon is inconstant and localized to the tip of the affected finger. When clinical and standard radiological examinations are normal, arteriography may be useful to confirm the diagnosis and locate the lesion. The main differential diagnoses before arteriography are post-traumatic neuroma and subungual melanoma. Surgical treatment is straightforward, but pain may recur post-operatively; it may be due to incomplete excision, development of a new tumor, presence of other tumors or cicatricial neuroma. PMID- 2988097 TI - Sympathetically induced changes in the responses of slowly adapting type I receptors in cat skin. AB - The effects of sympathetic efferent activity on slowly adapting Type I receptors in the hairy skin of cats were studied by recording from single afferent units in the saphenous nerve. Stimulation of the sympathetic trunk at 10 Hz had predominantly excitatory effects, which were seen in some units as the development of a background discharge in the absence of overt mechanical stimulation, or in most other units as a reduction in the threshold for mechanical activation. These effects generally persisted throughout the 3-min period of sympathetic stimulation (SS). The percentage of afferent units that began to discharge during SS was significantly greater in female cats than in males (53% vs. 19%). An increase in the force exerted by the skin on the stimulus probe was also observed during SS. Several tests were conducted to assess possible neurochemical and mechanical mechanisms of action. Administration of the alpha-adrenergic blocking agent phentolamine produced a marked reduction in the sympathetic effects. However, histochemical analysis of sections from the touch domes showed no catecholamine fluorescence near the sensory fibers. Cessation of local blood flow just prior to SS, produced by occlusion of the descending aorta, had no apparent effect on the sympathetically induced changes in afferent activity. It was concluded that sympathetic activity has an excitatory action on most Type I afferents in the cat. Because this sympathetic action is neither replicated nor altered by aortic occlusion, it appears not to be mediated by changes in blood flow. It also appears not to be mediated by direct neurotransmitter action on the sensory receptor, because no catecholamine fluorescence was observed, yet the action was blocked by an alpha-adrenergic blocker. It is likely, therefore, that this sympathetic action is mediated by some unidentified mechanical response within the skin. PMID- 2988099 TI - Is insulin-dependent diabetes mellitus caused by coxsackievirus B infection? A review of the epidemiologic evidence. AB - The evidence that coxsackievirus B plays a causal role in the etiology of insulin dependent diabetes mellitus (IDDM) is reviewed. This hypothesis is biologically plausible; one variant of another picornavirus, encephalomyocarditis virus, causes diabetes in genetically susceptible mice, but prior infection with another, serologically indistinguishable variant prevents this. The seasonal distributions of infection due to coxsackievirus B and of IDDM are similar. Case reports document coxsackievirus B infection coincident with the onset of IDDM. More than one-third of patients with recent-onset IDDM have antibody to coxsackievirus B but have not necessarily had recent infection. Case-control studies show no consistent association, with odds ratios ranging from 0.7 to 20. In two prospective studies, IDDM did not follow coxsackievirus B epidemics, but the sample sizes were small. Although epidemiologic data are too inconsistent to allow one to conclude that coxsackievirus B is a frequent cause of IDDM, methodologic difficulties--illustrated in the animal model--suggest that it would be premature to discard this hypothesis. PMID- 2988101 TI - Management of the deficient alveolar ridge: hydroxylapatite augmentation. PMID- 2988098 TI - Transmission of viral infections by the water route: implications for developing countries. AB - The "enteric" virus group comprises greater than 100 different viruses. These viruses typically infect the cell lining of the alimentary canal and are discharged in very large numbers in the feces of infected persons. Contamination of water supplies by enteric viruses represents an important source of viral infection. Many communities, particularly in developing countries, depend on sewage-polluted sources for their recreational and drinking water. Because conventional methods of sewage and water treatment have proved inefficient in the removal and inactivation of most enteric viruses, great concern has been raised over the impact of waterborne infection on the health of such communities. Current evidence implicating drinking and recreational water supplies in the transmission of nonbacterial gastroenteritis and hepatitis A virus and adenovirus infections is overwhelming. Water-borne transmission of other enteric viruses is also possible. Effective antiviral drugs are generally unavailable, and current vaccines can control only a limited number of viral infections; therefore, provision of uncontaminated water is a basic requirement in raising the standard of health in affected communities. PMID- 2988100 TI - [Different models for the study of a new anti-asthmatic substance]. AB - The various physiopathological components of asthmatic disease are interwoven. Experimental models used to study the sites of impact of an anti-asthmatic substance seek to separate the different mechanisms. The in vitro bronchial tissue model is rarely available in man. Progress is being made towards the development of a multicellular and membrane experimental model. However, it is still difficult to establish links between clinical findings, the results of respiratory function tests and biological results at cell or membrane level. There would seem to be a number of essential basic factors in this area: determination of the categories of asthma studied, regular surveillance of respiratory function tests combining spirometric and plethysmographic studies with pharmacodynamic tests, precise therapeutic protocols and the simultaneous use of several cellular biological models. PMID- 2988103 TI - [Pains of deafferentiation]. PMID- 2988102 TI - Role of retinoids and their binding proteins in filarial parasites and host tissues. PMID- 2988104 TI - Estimation of stability of [3H]-ouabain binding site concentration in rat and human skeletal muscle post mortem. AB - The post mortem stability of the [3H]-ouabain binding site concentration and 3-O methylfluorescein phosphatase (MFPase) activity was evaluated in rat skeletal muscle. As compared with the values measured in fresh tissue, the [3H]-ouabain binding site concentration in rat soleus muscle only dropped by around 1% per hour post mortem and a significant decrease was only seen after 12 h (15%, p less than 0.02). The 3-O-MFPase activity in rat gastrocnemius muscle showed a similar decrease. After 4 days, both parameters had dropped by 65% (p less than 0.001). In contrast, when intact fresh rat soleus muscles were incubated in Krebs-Ringer bicarbonate buffer at 20 degrees C for 4 days no significant decrease was seen in the [3H]-ouabain binding site concentration. In 10 human subjects the concentration of [3H]-ouabain binding sites was measured in specimens of the vastus lateralis muscle obtained within half an hour and at 6 and 12 h post mortem. The relative decrease after 6 h was insignificant (8%, p less than 0.30), whereas it was significant after 12 h (29%, p less than 0.005). These results have shown that the [3H]-ouabain binding sites in human skeletal muscle are resistant to post mortem degradation during the first 6 h after death. This makes it possible to perform measurements post mortem of the [3H]-ouabain binding site concentration in human skeletal muscle. PMID- 2988105 TI - Urinary cyclic AMP in spot urine of healthy children. AB - We present reference values for the excretion of cAMP in spot urine collected between 09.00 and 12.00 hours in 143 healthy children aged 2-200 months. The excretion of cAMP was creatinine-corrected and expressed as a substance concentration ratio (UcAMP/crea)U due to a positive significant correlation between the excretion of cAMP and creatinine (r = 0.68, p less than 0.001). The mean value (95% significance limits) for (UcAMP/crea)U was 748 mumol/mol (254 2206 mumol/mol). A logarithmic transformation of the ratio was used, since preliminary analysis showed uneven distribution; when the logarithmic transformation was used, the data appeared evenly distributed. There was no significant difference between the results for girls and boys. The value of lg(UcAMP/crea)U was related to the age or body surface area with decreasing values at higher age or body surface area. (r = -0.55 and r = -0.57, p less than 0.001). Spot urine for measurement of urinary cAMP instead of a 24 h collection appears preferable due to the practicability of the test in children. PMID- 2988106 TI - Arginine vasopressin and cyclic adenosine monophosphate during acute sodium loading in chronic glomerulonephritis. AB - The relationship of arginine vasopressin (AVP) in plasma to cyclic adenosine 3' 5'-monophosphate (cAMP), sodium excretion in urine, and arterial blood pressure were determined during intravenous infusion of hypertonic sodium chloride solution (500 ml of 50 g/l) in 10 normotensive control subjects and in 11 normotensive and 10 hypertensive patients with chronic glomerulonephritis and relatively well preserved kidney function. The concentration of AVP in plasma increased 2-4 fold, osmolality in serum increased 12-16 mosmol/kg, and urinary excretion of cAMP increased 20-40% during sodium loading to the same extent in all three groups. Sodium and water excretion were higher during the sodium loading in the hypertensive patients, but not in the normotensive patients when compared to the control subjects. Neither AVP nor changes in AVP correlated significantly with changes in cAMP excretion, sodium excretion or blood pressure. In the control subjects the level of parathyroid hormone in serum was unchanged during the sodium chloride infusion. Water loading without sodium loading in eight of the control subjects caused a decrease in the excretion of cAMP. In conclusion, the increase in cAMP excretion in urine during the sodium loading might be explained by an AVP-induced stimulation of renal cAMP production. The study does not suggest that AVP plays a role in the increased sodium excretion during sodium loading or in the development of hypertension or chronic glomerulonephritis. PMID- 2988107 TI - The effect of coarse wheat bran in the irritable bowel syndrome. A double-blind cross-over study. AB - Dietary supplementation with wheat bran has been widely advocated as a first-line treatment of patients with the irritable bowel syndrome (IBS). Few controlled trials have been reported, and the results are, furthermore, contradictory. The present study comprised 20 patients with IBS, of whom 18 (14 women, 4 men) completed the trial. The two treatment periods of 6 weeks each, with a daily intake of 30 g coarse wheat bran or 30 g placebo bran, respectively, were randomized in a double-blind cross-over design. Wheat bran significantly (P less than 0.05) increased the stool weight and shortened the intestinal transit time but was without significant effect on the colonic motility index was shown. We conclude that coarse wheat bran used as the only treatment in IBS does not provide a sufficient effect in a 6-week period. However, wheat bran seems to be justified in the treatment of constipation. PMID- 2988108 TI - The effect of guar gum and fiber-enriched wheat bran on gastric emptying of a semisolid meal in healthy subjects. AB - The effect of physiological doses of guar gum (Guarem), 5 g, and fiber-enriched wheat bran (Fiberform), 10.5 g, on gastric emptying was studied by two different methods in healthy subjects: by a simple isotope localization monitor placed over the upper part of the abdomen and by gamma camera. The fiber preparations were added to a semisolid meal consisting of wheatmeal porridge and juice, using technetium-99 DTPA as a marker. The gamma camera showed no effect of fiber on gastric emptying. The isotope localization monitor, however, indicated that Fiberform prevented a postprandial accumulation of the meal within the upper part of the stomach. The simple isotope localization monitor cannot be recommended for measurements of gastric emptying. PMID- 2988109 TI - The mechanism of action of omeprazole--a survey of its inhibitory actions in vitro. AB - In order to study the mechanism of inhibition of gastric acid secretion by omeprazole, its action was investigated in several different in vitro preparations. In preparations from isolated gastric mucosa, isolated rabbit gastric glands and isolated parietal cells, omeprazole was found to inhibit both basal and stimulated acid secretion. These effects were seen irrespective of whether acid formation was stimulated by histamine or by db-cAMP. The inhibitory pattern of omeprazole was found to be of a non-competitive nature against db-cAMP stimulation. Furthermore, in isolated glands, omeprazole was found to inhibit stimulation induced by high medium K+ and low Na+ concentrations. The basal membrane of the intact gland preparation was made permeable to molecules of large size by the use of digitonin, and acid secretion was subsequently initiated by the addition of exogenous ATP. Even under these conditions, omeprazole was found to be inhibitory, with an IC50-value comparable to that of intact glands. SCN- was found to mimic the action of omeprazole in that it counteracted both basal and stimulated acid secretion in the test models described above. In contrast, cimetidine was found to inhibit only histamine stimulation, consonant with its H2 receptor-blocking properties. In the gastric gland preparation, changes in oxygen consumption is closely related to changes in acid formation. When oxygen consumption and acid formation were measured in parallel under histamine stimulation, another benzimidazole, timoprazole (H 83/69) (structurally related to omeprazole), was found to inhibit both parameters. However, under non stimulated conditions, timoprazole was found to have only a minor effect on the oxygen consumption. The isolated H+K+ATPase preparation was used in order to investigate the effects of omeprazole at the "proton pump level". This enzyme was found to be inhibited by omeprazole in a pH-dependent manner. Under neutral or slightly alkaline conditions, slight inhibition occurred. When the pH of the incubation media was progressively decreased, the inhibitory activity of omeprazole was augmented. Several reactions of the H+K+ATPase enzyme cycle were investigated, i.e., K+-stimulated ATPase- and pNPPase- activities and formation of phosphoenzyme. All three of these reactions were inhibited. The results presented are in agreement with the hypothesis that omeprazole inhibits gastric acid secretion by blocking the gastric H+K+ATPase. PMID- 2988110 TI - Emerging strategies in ulcer therapy: pumps and receptors. AB - The treatment of gastric and duodenal ulcers has recently been revolutionised. The reason for this is the increased knowledge of the physiology and biochemistry of the mucosae at the cellular and subcellular levels. In this article, we try to explain how the gastric parietal cell works, and how, based on that knowledge, we might be able to devise increasingly sophisticated tools for inhibition of acid secretion. PMID- 2988111 TI - Transferrin receptors on circulating monocytes in hereditary haemochromatosis. AB - In patients with hereditary haemochromatosis (HH) abnormal functional properties of the macrophage system have been observed. The present study is a preliminary report of increased transferrin receptor expression on monocytes from 12 patients with HH. There was no correlation between the degree of iron overload and the transferrin receptor expression on the monocytes. The results obtained thus indicate that the observed increase in transferrin receptors is not a secondary phenomenon due to systemic iron overload but could be an expression of a primary inborn error of iron metabolism in HH. The functional aspects of the receptors were not evaluated as they were analyzed by means of monoclonal antibody technique. PMID- 2988112 TI - Plasma cyclic nucleotide levels in patients with homozygous beta-thalassaemia. AB - To investigate the possibility that a proliferative non-neoplastic process influences extracellular cyclic nucleotide concentrations, we measured plasma cyclic AMP and cyclic GMP levels in 38 patients with homozygous beta thalassaemia. This group consisted of 20 patients with thalassaemia major transfused regularly (mean pre transfusion Hb levels, 11 g/dl), and 18 patients with thalassaemia intermedia who did not require regular blood transfusion (mean Hb levels, 8.7 g/dl). In the patient group, plasma cyclic AMP levels were similar to those of 37 normal subjects matched for age and sex, whereas plasma cyclic GMP levels were markedly higher. Moreover, in the thalassaemic patients there was a significant negative correlation between plasma cyclic GMP levels and haemoglobin concentrations, suggesting that their marked erythroid hyperplasia may play a role in determining alterations in extracellular cyclic GMP levels. PMID- 2988113 TI - Autoimmune antigens and viral infection. PMID- 2988114 TI - Synovial fluid ferritin in rheumatic diseases. AB - The synovial fluid ferritin level in 49 patients (57 joints) with various rheumatic diseases was analysed. In rheumatoid arthritis (n = 22) the geometric mean ferritin level was 528 micrograms/l (range 56-3 100 micrograms/l), in other inflammatory arthritides (n = 12) 339 micrograms/l (105-2 835 micrograms/l) (p greater than 0.5), in calcium pyrophosphate arthropathy (n = 14) 507 micrograms/l (180-4 230 micrograms/l) (p greater than 0.5) and in non-inflammatory osteoarthritis (n = 9) 167 micrograms/l (14-725 micrograms/l) (p less than 0.05). Synovial fluid/serum ferritin ratios did not differ significantly in the four diagnostic groups; 4 patients had ratios less than 1.0. Synovial fluid ferritin was not correlated to total fluid cell count or differential cell count. Although ferritin content was significantly greater in inflammatory than in noninflammatory fluid (p less than 0.05), the wide scatter of the values and marked overlap between the different groups limit the value of measuring synovial fluid ferritin as a differential diagnostic test for rheumatic diseases. PMID- 2988115 TI - [Occult gastric adenocarcinoma with pulmonary carcinomatous lymphangitis and microangiopathic hemolytic anemia in a young adult]. AB - The case is reported of a 19-year-old patient with gastric carcinoma, in which clinical presentation (intermittent fever, myalgia, proximal muscle weakness and diffuse nodular-trabeculated infiltration of both lungs) was very unusual. The patient developed further complications (microangiopathic hemolytic anemia with disseminated intravascular coagulation) and died of subdural hematoma. Bone marrow biopsy showed metastatic mucin-producing adenocarcinoma, but the gastric primary site of the tumor could only be demonstrated at autopsy. PMID- 2988116 TI - [Potassium regulation--current understanding from a clinical viewpoint]. AB - Mechanisms of renal potassium excretion and internal potassium balance as the main determinants of serum and/or total body potassium are discussed. Renal handling of potassium is reviewed, with particular attention to the effects of diuretics, acid-base disturbances and aldosterone-deficient states. Among the regulatory processes of internal potassium balance, the importance of adrenergic stimuli/drugs, of acid-base balance, and of magnesium and insulin is stressed. Physiologic and pathophysiologic evidence provides the basis for discussion of relevant clinical implications. PMID- 2988117 TI - [Acquired immunodeficiency syndrome (AIDS). An update after 4 years]. AB - At the end of December 1984, 8246 AIDS cases had been reported in the United States, and 762 cases in Europe. The main risk groups are homosexual and bisexual men (73%), heterosexual i.v. drug addicts (17%) and Haitians (4%). AIDS is diagnosed in patients meeting the clinical criteria of the CDC surveillance definition (opportunistic infections and/or Kaposi's sarcoma). The prevalence of HTLV-III antibodies in risk groups is high, its prognostic significance still controversial. Laboratory methods to determine the more relevant circulating viral proteins or the whole virus are now being developed. Limited studies to evaluate the possibility of reconstituting the immune system of AIDS patients with interleukin-2 and gamma-interferon have proved unsuccessful. successful. Several new drugs with possible antiviral properties are now being evaluated in first clinical trials. PMID- 2988118 TI - [Autonomic nerve activity and ventricular fibrillation threshold]. PMID- 2988119 TI - [Effect of peptides on thermoregulation]. PMID- 2988120 TI - [New aspects of the mechanism of action of hormones containing nitrogen]. PMID- 2988121 TI - [Impulsive activity not the sole neural determinant of the type of skeletal muscle fiber]. PMID- 2988122 TI - [Metabolism and its hormonal regulation in short- and long-distance competitive swimming]. PMID- 2988124 TI - Retinal S antigen identified as the 48K protein regulating light-dependent phosphodiesterase in rods. AB - Retinal S antigen chromatographically purified from whole retina, induces experimental autoimmune uveoretinitis in laboratory animals. The 48K protein, a soluble protein found in rod outer segments, is purified through its specific binding to photoexcited rhodopsin and is involved in the quenching of light induced guanosine 3',5'-monophosphate-phosphodiesterase activity. Biochemical, immunological, functional, and pathological tests showed that retinal S antigen and the 48K protein are identical. PMID- 2988123 TI - The LDL receptor gene: a mosaic of exons shared with different proteins. AB - The multifunctional nature of coated pit receptors predicts that these proteins will contain multiple domains. To establish the genetic basis for these domains (LDL) receptor. This gene is more than 45 kilobases in length and contains 18 exons, most of which correlate with functional domains previously defined at the protein level. Thirteen of the 18 exons encode protein sequences that are homologous to sequences in other proteins: five of these exons encode a sequence similar to one in the C9 component of complement; three exons encode a sequence similar to a repeat sequence in the precursor for epidermal growth factor (EGF) and in three proteins of the blood clotting system (factor IX, factor X, and protein C); and five other exons encode nonrepeated sequences that are shared only with the EGF precursor. The LDL receptor appears to be a mosaic protein built up of exons shared with different proteins, and it therefore belongs to several supergene families. PMID- 2988125 TI - Expression of Plasmodium falciparum circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine. AB - The circumsporozoite (CS) protein of the human malaria parasite Plasmodium falciparum may be the most promising target for the development of a malaria vaccine. In this study, proteins composed of 16, 32, or 48 tandem copies of a tetrapeptide repeating sequence found in the CS protein were efficiently expressed in the bacterium Escherichia coli. When injected into mice, these recombinant products resulted in the production of high titers of antibodies that reacted with the authentic CS protein on live sporozoites and blocked sporozoite invasion of human hepatoma cells in vitro. These CS protein derivatives are therefore candidates for a human malaria vaccine. PMID- 2988127 TI - Deregulation of interleukin-2 receptor gene expression in HTLV-I-induced adult T cell leukemia. AB - Infection of human T cells by human T-lymphotropic virus, type I (HTLV-I), a retrovirus, is uniformly associated with the constitutive expression of large numbers of cellular receptors for interleukin-2 (IL-2). Comparison with normal T cells shows that neither IL-2 receptor gene organization nor IL-2 receptor messenger RNA processing are altered in the leukemic cells. However, mitogenic stimuli activate IL-2 receptor gene expression in normal T cells, whereas these stimuli paradoxically inhibit IL-2 receptor gene transcription in HTLV-I-infected leukemic T cells. PMID- 2988126 TI - Immunogenicity of synthetic peptides from circumsporozoite protein of Plasmodium falciparum. AB - In a study of recombinant proteins that might be useful in developing a vaccine against malaria, synthetic peptides from the circumsporozoite (CS) protein of Plasmodium falciparum were found to be immunogenic for mice and rabbits. Antibody to peptides from the repeating region of the CS protein recognized native CS protein and blocked sporozoite invasion of human hepatoma cells in vitro. Antibodies to peptides from regions I and II had no biologic activity, although antibody to region I recognized processed CS protein by Western blot analysis. These data support the feasibility of developing a vaccine against the sporozoite stage of the malaria parasite by using synthetic peptides of the repeating region of the CS protein conjugated to a carrier protein. PMID- 2988128 TI - Biochemical modeling of an autonomously oscillatory circadian clock in Euglena. AB - Eukaryotic microorganisms, as well as higher animals and plants, display many autonomous physiological and biochemical rhythmicities having periods approximating 24 hours. In an attempt to determine the nature of the timing mechanisms that are responsible for these circadian periodicities, two primary operational assumptions were postulated. Both the perturbation of a putative element of a circadian clock within its normal oscillatory range and the direct activation as well as the inhibition of such an element should yield a phase shift of an overt rhythm generated by the underlying oscillator. Results of experiments conducted in the flagellate Euglena suggest that nicotinamide adenine dinucleotide (NAD+), the mitochondrial Ca2+-transport system, Ca2+, calmodulin, NAD+ kinase, and NADP+ phosphatase represent clock "gears" that, in ensemble, might constitute a self-sustained circadian oscillating loop in this and other organisms. PMID- 2988130 TI - Radionuclide imaging of parathyroid tumors: historical perspectives and newer techniques. AB - The increasing use of automated blood chemistry screens for serum calcium levels along with improved methods in measuring parathyroid hormone (PTH) levels have made the diagnosis of parathyroid disease a common clinical problem. Parathyroid adenomas account for the majority of primary hyperparathyroidism with diffuse hyperplasia and parathyroid carcinoma occurring less frequently. Early scintigraphic techniques to identify enlarged parathyroids used selenomethionine 75 which was considered to be incorporated into PTH. In general, the sensitivity of scanning the neck using this tracer was related to the size of the enlarged parathyroid, but in large series, the overall sensitivity was less than 50%. Recent work by Ferlin et al, using a Technetium-99m/Thallium-201 subtraction scintigraphic technique has yielded a sensitivity of 92% in identifying pathologically enlarged parathyroid glands. Winzelberg et al modified this technique to allow imaging the mediastinum plus simplifying the subtraction method. In a prospective study with high-resolution sonography, similar sensitivities and specificities were found with sonography and scintigraphy. Tl 201/Tc-99m pertechnetate subtraction scintigraphy appears to be an accurate technique in identifying pathologic parathyroid enlargement. Its ultimate role in the evaluation of patients with suspected hyperparathyroidism still needs to be determined. PMID- 2988129 TI - Radionuclide diagnosis and therapy of thyroid cancer: current status report. AB - Thyroid cancer is uncommon, with an incidence of 10,300 new patients each year and a mortality of 1,100 patients each year. Patient survival correlates with many factors, including tumor pathology, age, primary lesion size, distant metastases, extent of surgery, and radioiodine therapy. Deaths from thyroid cancer may occur many years after diagnosis, and such an indolent course has hampered the analysis of the multiple treatment programs advocated. Thyroid imaging continues to play an important role in the initial detection and follow up management of thyroid cancer, but the search for a specific tracer for the primary lesion continues. The complementary role of serum thyroglobulin and radioiodine in the follow-up of the thyroidectomized patient is discussed. Radioiodine therapy has proven effectiveness in those patients with radioiodine avid distant metastases and/or regional metastases. Whether radioiodine ablation of residual thyroid bed activity is beneficial remains controversial. PMID- 2988131 TI - Pediatric applications in nuclear oncology. AB - Imaging procedures (both as they apply to initial diagnosis and assessing the response to therapy) using radionuclides are indispensible in the evaluation of malignant diseases that afflict infants and children. The diagnosis and accurate staging are extremely critical because of continuing improvements in the survival of children with malignant disease when treated with appropriate therapy. The major role of nuclear medicine in this patient group serves to evaluate the skeletal system and the liver and spleen. It is important to keep the pediatric specialist well informed concerning the role that nuclear medicine can play in elucidating many of the clinical problems that this specialist faces. PMID- 2988132 TI - Lymphocyte markers and infectious diseases. PMID- 2988133 TI - [The alcoholic liver: steatosis, hepatitis, cirrhosis, cancer]. PMID- 2988134 TI - [Natural history of cirrhosis. The 4 revealing complications]. PMID- 2988135 TI - Amplification and expression of phenylalanine hydroxylase in mouse erythroleukemia cells. AB - A cell line, CY-1, was selected in tyrosine free (tyr-) medium after fusion of mouse erythroleukemia (MEL) cells with mitomycin C-treated rat hepatoma cells. MEL cells do not express the enzyme phenylalanine hydroxylase (PH) and are unable to grow in tyr- medium, whereas the rat hepatoma cells constitutively express PH and are able to grow in tyr- medium. CY-1 cells resemble MEL cells morphologically, karyotypically, and in being inducible for hemoglobin synthesis. In contrast to MEL cells, CY-1 expresses PH and is therefore able to grow in tyr- medium. Using a rat cDNA probe for the PH gene, Southern blot analyses were carried out on DNA isolated from CY-1 and parental cells. CY-1 showed the characteristic mouse PH gene pattern but the gene copy number was amplified four- to eightfold compared to parental MEL cells. PMID- 2988136 TI - Shuttling of integrated vectors from mammalian cells to E. coli is mediated by head-to-tail multimeric inserts. AB - With the aim of producing nonviral shuttle vectors for mammalian cells, we have constructed mouse mitochondrial DNA derivatives comprising the xanthine-guanine phosphoribosyltransferase gene as a selectable marker. Complete or subcomplete mitochondrial genomes were inserted into the plasmid pBB3 and transferred into hepatoma cells in order to generate, in vivo, new recombinant molecules. A second and a third-generation vector, p12.2b and p delta respectively, were thus isolated for their ability to shuttle from mammalian cells to recA+ E. coli. Transfection of rodent fibroblasts and hepatoma cells showed that, contrary to our expectations, p12.2b and p delta are not self-replicating episomes; their shuttling from mammalian cells to recA+ E. coli is mediated by tandem integrated copies. The relevant property of p12.2b and p delta is a ubiquitous propensity to form head-to-tail multimeric structures when they integrate into mammalian host chromosomes. This ability is missing in pBB3 and appears only following the insertion of various mitochondrial or nuclear DNA fragments into the plasmid. These data are discussed in terms of homologous recombination and shuttling of integrated vectors. PMID- 2988137 TI - Molecular approach to analyzing the human 5p deletion syndrome, cri du chat. AB - DNA unique or low-copy fragments were isolated from a genomic DNA library specific for the short (p) arm of human chromosome 5. These chromosome 5p specific DNA fragments were used to analyze, by Southern blot experiments, somatic cell hybrids that retained either a normal chromosome 5 homolog or a homolog with a partial deletion of 5p, which was derived from either of two persons with the common human deletion syndrome, cri du chat or 5p- syndrome. In these studies, two classes of DNA fragments were identified, those located outside the region deleted in the persons with cri du chat and those located within the deleted region. This latter class of DNA probes will help to define, at the molecular level, a region of 5p that is critical in producing the phenotype associated with the cri du chat syndrome. PMID- 2988138 TI - Expression of the receptor for epidermal growth factor correlates with increased dosage of chromosome 7 in malignant melanoma. AB - Epidermal growth factor (EGF) receptor is expressed selectively by human melanoma cells which show the presence of an extra copy of chromosome 7. None of the cells of benign pigmented lesions (nevi) or radial growth phase (nonmetastatic) primary melanoma expressed EGF receptor and none of these cells showed an extra copy of chromosome 7. The results indicate that a single extra dose of a gene (for EGF receptor) may provide a selective advantage to cells in the late stages of tumorigenesis. PMID- 2988139 TI - [Percutaneous transhepatic biliary drainage in malignant extrahepatic cholestasis]. PMID- 2988140 TI - [Diseases in the area of the petrous bone: the value of high-resolution computed tomography]. PMID- 2988141 TI - Prevalence of antibodies to varicella zoster virus in healthy adults. AB - A serological study of immunity to varicella zoster was carried out using an enzyme-linked immunosorbent assay (ELISA) in 244 healthy adult laboratory staff members. The overall immunity was 90%, with a progressive increase from 81% at 20 29 years to 100% at 60 years. Approximately one-third of serologically immune individuals had no certain history of varicella. As the serological test is simple, rapid and reliable, screening for immunity should be carried out in at risk individuals such as immunosuppressed patients, pregnant women, and laboratory and medical patients, pregnant women, and laboratory and medical personnel who come into contact with sources of virus. Definition of serological status will aid in the rational planning of intervention procedures such as epidemiological control and administration of varicella zoster immune globulin and, when it becomes available, varicella vaccine. PMID- 2988142 TI - Sero-epidemiology of HTLV-III antibody in southern Africa. AB - A preliminary survey has demonstrated that antibodies directed against human T cell leukaemia virus type III (HTLV-III), the virus implicated as the agent causing the acquired immunodeficiency syndrome, are not present in the low-risk population groups studied. The survey, in which an indirect immunofluorescence assay was used, has indicated that HTLV-III is not endemic in southern Africa (as opposed to central Africa, where it has been suggested that the virus is endemic). Anti-HTLV-III antibodies were, however, found in sera from male homosexuals, the one high-risk population group studied. PMID- 2988144 TI - Prevalence of serum antibodies to herpes simplex virus types 1 and 2: application of an ELISA technique to 100 cases of anogenital herpes. AB - Sera of 100 patients affected by anogenital herpes were tested by an enzyme linked immunosorbent assay (ELISA) technique using partially purified antigens obtained by extraction from the nuclei of infected Vero cells. Herpes simplex virus type 1 (HSV-1) was isolated from the anogenital lesions of 17 patients and herpes simplex virus type 2 (HSV-2) from 83 patients. The relative proportions of antibody of the IgG class to HSV-1 and HSV-2 were determined and their ratio (R) was calculated, except for 23 patients for whom the corrected optical density (COD) obtained for HSV-1 or HSV-2 antibody was less than the lower value limit of 0.025. A predominance of HSV-2 antibody (R less than 1) was found in 40 patients and was uniformly associated with isolation of HSV-2, whereas a predominance of HSV-1 antibody (R greater than 1) in 37 patients was found in the presence of infection with HSV-1 in 11 (30%) and with HSV-2 in 26 (70%) patients. This study also confirms the association of a higher frequency of recurrences with HSV-2: among the 55 patients with recurrent herpes, those from whom HSV-2 was isolated had a history of more recurrences than those infected with HSV-1 (P less than 0.05; Mann and Whitney's nonparametric test). No correlation between the number of recurrences and the level of HSV-2 antibody was found. PMID- 2988143 TI - Frequency of acquisition of first-episode genital infection with herpes simplex virus from symptomatic and asymptomatic source contacts. AB - Sixty-six source contacts of index patients with first-episode genital infection caused by herpes simplex virus (HSV) were evaluated for evidence of current or past HSV infections. Forty-three source contacts (65%) reported a history consistent with previous recurrent HSV infection or were experiencing a first episode of genital herpes when initially examined. However, 60% of these 43 contacts were not aware that they had transmissible HSV infection. Twenty-nine (67%) of the 43 individuals had had recent sexual contact with an index patient when lesions were present. All of the remaining 23 source contacts, who were without a history of symptoms consistent with HSV infection, had detectable neutralizing antibody to HSV; HSV type 2 was isolated from the cervix of two of these asymptomatic source contacts. Efforts to identify individuals with undiagnosed genital herpes and to instruct these individuals concerning the risk of disease transmission in the presence of lesions are needed if the rate of transmission is to be decreased; however, methods designed to decrease the rate of transmission by asymptomatic individuals must also be evaluated. PMID- 2988145 TI - [Value of biologic tests in breast cancer]. PMID- 2988147 TI - Leiomyomatosis peritonealis disseminata. AB - Leiomyomatosis peritonealis disseminata is a rare benign disorder in females characterized by multiple smooth muscle tumor nodules throughout the peritoneal cavity. All of the reported cases have been in menstrual-age women; some of whom have been pregnant or taking oral contraceptives. We report a postmenopausal woman who had symptoms related to bowel-wall involvement in two episodes 12 years apart. PMID- 2988148 TI - [Calcium pyrophosphate crystal deposition disease]. PMID- 2988146 TI - Lambert-Eaton myasthenic syndrome. AB - The Lambert-Eaton myasthenic syndrome is associated in about 65% of cases with small cell carcinoma, a tumour of neurosecretory origin. It is characterised physiologically by a decrease in the nerve evoked quantal release of acetylcholine, and in the resting non-quantal release ("molecular leakage"). The associations with autoimmune disease, with other autoantibodies, with HLA-B8, and with the IgG heavy chain marker Glm (2) are consistent with an autoimmune aetiology. Clinical and electromyographic responses to plasma exchange point to a humorally mediated disorder. This has been substantiated by passive transfer of the the main electrophysiological features of LEMS to mice by daily injections of LEMS IgG. Plasma was no more effective in inducing the electrophysiological changes than the IgG fraction. The decrease in quantal content appeared closely to follow the level of human IgG in the mouse serum and complement (C5) deficient mice were as susceptible as normal controls. The principal physiological abnormalities are both Ca2+ dependent processes, suggesting that a defect in Ca2+ transport may underlie the disorder. Preliminary studies of quantal content at low Ca2+ concentrations in mice injected with LEMS IgG suggest the functional loss of 40% of Ca2+ channels. Electron microscopic freeze fracture studies in such animals show, as in the human disease, a significant reduction in the number of active zone particles which are believed to represent Ca2+ channels. Thus it seems likely that the disorder of acetylcholine release is due to an IgG antibody directed to nerve terminal determinants that include the Ca2+ channels or structures closely related to them. In cancer-associated LEMS, the autoantibody response may initially be made to similar determinants on the tumour cell membrane, cross-reactivity of the antibody with nerve terminal determinants leading to the disorder of transmitter release. PMID- 2988149 TI - [Functional state of the pituitary-adrenal system in patients with nonspecific ulcerative colitis]. PMID- 2988150 TI - [Radionuclide diagnosis of chronic pancreatitis]. PMID- 2988151 TI - [The effect of beclomethasone dipropionate as inhalation therapy on the pituitary adrenal axis in children]. AB - The function of the pituitary-adrenal axis was investigated in ten asthmatic children (5-14 years of age) before and after administration of beclomethasone dipropionate as inhalation therapy in a dose of 300-600 micrograms/24 hrs, during six months. Cortisol diurnal rhythm, excretion of free cortisol in 24 hours urine and stimulation of plasma-cortisol after ACTH-administration were determined. The same investigations were carried out in a comparable group of ten asthmatic children already using this inhalation therapy for more than one year. Significant differences were found comparing 08.00 h. cortisol values of the children from the first group and those who used medication for more than one year. In this first group, the ability of the adrenal glands to be stimulated by ACTH-administration after six months of medication, decreased significantly as well. These results indicate an impairment of the function of pituitary-adrenal axis during long-term treatment with inhaled beclomethasone-dipropionate. This implicates a decreased ability of the adrenal glands to react in an optimum way in stress-situations. In these patients oral or parenteral administrations of steroids should be considered earlier in stress situations like a severe asthma attack. PMID- 2988152 TI - Different functions and associations of HLA-DR and HLA-DQ(DC) antigens shown by serological, cellular and DNA assays. AB - Two consanguineous Caucasoid HTCs, DHI and FPA, were investigated, the latter having an unusual HLA-DR/DQ(DC) association. Both these HTCs typed clearly as HLA DRw11. However, while DHI typed as DRw11/DQw3(DC4) as expected, FPA typed as DRw11/DQw1(DC1) instead. Although extremely rare in Caucasoids, DRw11/DQw1 is a common pattern of association in Nigerian Negroids. Southern blots of DNA extracted from EBV cell line derived from FPA, hybridized with HLA-DC alpha and HLA-DC beta probes, confirmed this unusual DRw11/DQw1(DC1) association. In addition the DC alpha probe showed a unique additional restriction fragment length polymorphism (8 kb) attributable to the DX gene in the FPA DNA. When DHI and FPA were used as stimulators in MLC, the patterns to typing responses obtained were not completely concordant although they overlap to some extent. For this reason FPA has been locally designated Dw'F5', distinct from Dw5. Furthermore, the HLA-DQ antigens of the responder cells were not necessarily the same as those of the HTCS to which they gave typing responses (FPA and DHI). Functional studies using these two HTCs showed that the DQ(DC) antigens probably have no direct lymphocyte activating properties but rather have a regulatory role in controlling responses to allodeterminants in MLC. PMID- 2988153 TI - Leupeptin, a protease inhibitor, blocks insemination-induced flight muscle histolysis in the fire ant Solenopsis. AB - The effect of the protease inhibitor leupeptin on flight muscle histolysis in queen fire ants was studied by electron microscopy. In untreated animals artificially inseminated, muscle involution was apparent at 6 hr post insemination and complete by 24 hr post-insemination. However, in animals pre treated with leupeptin and subsequently artificially inseminated, no morphologic evidence of flight muscle breakdown was seen at any interval between 6 and 24 hr post-insemination. Such information appears to indicate that one or more proteases are involved in the process of insemination-induced muscle atrophy in fire ants. The most likely candidate is a soluble, calcium-activated myofilament associated protease. PMID- 2988154 TI - Comparison of the action of types A and F botulinum toxin at the rat neuromuscular junction. AB - Blockade of neuromuscular transmission was produced in the lower hind limb of the rat by local injection of either type A or type F botulinum toxin (BoTx). At 1, 3, 7, and 10 days after injection, the extensor digitorum longus (edl) nerve muscle preparation was excised and analyzed for alterations in muscle mechanical properties or spontaneous and nerve stimulus-evoked quantal transmitter release. Muscles receiving type A toxin were paralyzed up to and including 7 days after injection. Muscles treated with type F toxin, although completely paralyzed at 1 and 3 days after injection, twitched in response to nerve stimulation by 7 days. Both toxins induced a marked decrease in the frequency of miniature end-plate potentials, but type A did so to a greater extent. Between 1 and 3 days after toxin injection nerve impulse-evoked transmitter release was reduced in both type A- and type F-treated muscles. Evoked release was temperature sensitive in type A treated muscles but not in those treated with type F. 3,4-Diaminopyridine (3,4 DAP), a compound which increases nerve-evoked transmitter release by increasing Ca2+ influx, was more effective in reversing the paralysis in type A than in type F-treated muscles. 3,4-DAP induced asynchronous end-plate potentials in response to nerve stimulation in type F-paralyzed muscles, but not in muscles treated with type A. Amidination of the amino groups (presumably lysine) on the toxin by treatment with ethylacetimidate increased the potency and efficacy of only type F BoTx. The results show that type F BoTx differs from type A, mainly by its lower potency, efficacy, shorter duration of action, and by being less effectively antagonized by 3,4-DAP. PMID- 2988155 TI - Mechanism of chemical-induced toxicity. II. Role of extracellular calcium. AB - Previous studies disagree as to if chemical-induced cell death is caused by the influx and accumulation of extracellular Ca2+. To determine the role of extracellular Ca2+ in toxic cell death, the viability (leakage of intracellular K+ and lactate dehydrogenase) and total Ca2+ content of isolated hepatocytes incubated in the presence or absence of extracellular Ca2+ were determined during a toxic insult with bromobenzene, ethyl methanesulfonate (EMS), Ca2+ ionophore A23187, and adriamycin (ADR) in combination with 1,3-bis(2-chloroethyl)-1 nitrosourea (BCNU). The present study utilized the dibutyl phthalate separation technique which enabled the analysis of only viable hepatocytes for changes in intracellular Ca2+ and K+ content during toxic cell injury. The three chemical treatments, bromobenzene, EMS, and ADR-BCNU, each caused an accelerated loss of viability in hepatocytes incubated without extracellular Ca2+ as compared to cells incubated with Ca2+. Furthermore, the total Ca2+ content of viable hepatocytes incubated in the presence of extracellular Ca2+ did not increase during chemically induced cell injury as compared to control cells. In fact, a significant decline in total cellular Ca2+ was observed in viable hepatocytes incubated in Ca2+-free medium during toxic cell injury. Treatment with Ca2+ ionophore A23187 was also toxic to hepatocytes incubated in the presence or absence of extracellular Ca2+. At high concentrations of ionophore (20 microM or 4 micrograms/10(6) cells), cell death was accelerated in hepatocytes incubated with Ca2+ as compared to cells incubated in Ca2+-free medium. In contrast, after treatment with lower concentrations of ionophore (10 microM or 2 micrograms/10(6) cells), the rate of cell death was reversed with hepatocytes incubated without extracellular Ca2+ dying first. Thus, depending on the concentration of A23187 and the time of exposure, the presence of extracellular Ca2+ can be shown either to accelerate or protect against cell death. Surprisingly, reversible and irreversible cell injury were not observed in hepatocytes incubated with extracellular Ca2+ and 2 microM A23187 though this treatment resulted in an 800% increase in total intracellular Ca2+ content. We conclude that chemical-induced hepatic cell death is not caused by an increase in total cellular Ca2+ resulting from the influx of extracellular Ca2+. PMID- 2988156 TI - The modulation by arylamines of the in vitro formation of superoxide anion radicals and hydrogen peroxide by rat liver microsomes. AB - A concentration-dependent increase in the generation of the superoxide anion radical (O-2), was observed during the incubation of benzidine (0.1-5 mM), but not of the structurally related compounds 4-aminobiphenyl, 2-aminobiphenyl or 4 fluoro-4'-aminobiphenyl, with NADPH-supplemented rat liver microsomes. This increase was partially inhibited by carbon monoxide and catalase but unaffected by 4-aminobiphenyl, a substrate of the cytochrome P-450 system. Microsomes from benzo(a)pyrene-treated, but not microsomes from phenobarbitone-pretreated rats, were responsible for a larger benzidine-dependent effect compared to microsomes from control animals. In contrast to the above observations, benzidine decreased the formation of hydrogen peroxide by NADPH-supplemented microsomal preparations from untreated rats. These results indicate that free radicals of oxygen are generated during the metabolism of some arylamines. PMID- 2988157 TI - Activities of hepatic epoxide hydrolase and glutathione S-transferase in rats under the influence of tetramethyl thiuramdisulfide, tetramethyl thiurammonosulfide or dimethyl dithiocarbamate. AB - The epoxide hydrolase (EH) activity in the liver of adult female Wistar rats significantly increased 18 h after the administration by gavage of tetramethyl thiuramdisulfide (TMTD, 1 mmol/kg) or tetramethyl thiurammonosulfide (TMTM, 2 mmol/kg). No increase was observed 5 h after administration of Na-dimethyl dithiocarbamate (Na-DMDTC, 4 mmol/kg). The glutathione S-transferase (GST) activity in the cytosol and microsomes of the liver was slightly enhanced after oral (gavage) administration of TMTD, TMTM or Na-DMDTC (doses up to 4 mmol/kg). In vitro, TMTD, TMTM, and Na-DMDTC significantly enhanced the hepatic activity of EH prepared from adult female Wistar rats. Cytosolic and microsomal GST activities from the liver were significantly raised in vitro by Na-DMDTC. The results have a bearing on the evaluation of the risk to health of these chemicals in the workplace. PMID- 2988159 TI - The histopathology and electron microscopy of a human monkeypox lesion. AB - The histological and electron microscopic findings from a solitary cutaneous monkeypox lesion taken post mortem from a child who died after a five-day illness are reported. This child is 44th in the WHO register of monkeypox cases. The lesion was at the papulonecrotic stage, with early evidence of vesiculation and minimal evidence of pustulation. Necrosis affected the stratum basale, the related basement membrane and adjacent areas of the dermal papillae at the centre of the lesion. Cell necrosis affected the next two or three layers of stratum spinosum above the destroyed stratum basale. Lateral to this zone, marked hyperplasia and intracellular oedema of the stratum spinosum constituted the papule and produced spindle-cell features. In the middle layer of the stratum spinosum, above the necrotic focus, there were minute vesicles and between these were occasional multinuclear giant cells. Bodies similar to Guarnieri bodies (GB) were present in the cytoplasm of sweat duct-lining cells in the epidermis and upper corium. Very scanty similar bodies were evident elsewhere in the papular epidermis but were difficult to distinguish from debris. Granules in the lesion with the same size as mature virions (elementary bodies) have been assessed not to be these because similar granules are present in the normal epidermis. Changes in the dermis apart from those mentioned above were minimal oedema, very mild perivascular infiltration by round cells and an occasional eosinophil. Electron microscopy showed abundant immature and mature orthopoxvirus particles in the cytoplasms of infected epidermal cells. A limited range of histochemical tests is detailed. In general, the features are indistinguishable from the papulonecrotic stage of smallpox (variola) and from tanapox as recorded in man. PMID- 2988158 TI - A secondary prevention, randomized trial of suloctidil in patients with a recent history of thromboembolic stroke. AB - Four hundred and thirty-eight patients who had suffered a thromboembolic stroke not less than two weeks or more than four months previously, were entered into a placebo-controlled randomized clinical trial to determine whether suloctidil (200 mg t.i.d.) would influence the subsequent recurrence of stroke, the occurrence of myocardial infarction, or cardiovascular death. The two treatment groups were comparable at baseline with respect to important prognostic variables and there was good adherence to the study protocol during an average follow-up of 20 months. Significantly more patients complained of side-effects in the suloctidil group and more hepatotoxicity was also reported in the suloctidil group. Four cases of clinical hepatitis were suspected to be due to suloctidil, each of which was reversible on termination of study treatment; relative increases in SGOT and SGPT at three months in the suloctidil group were found to be mild and transient. The primary analysis of efficacy was based on the incidence of the first event of stroke, myocardial infarction or cardiovascular death, but excluding events that occurred more than 28 days after complete withdrawal from study medication for whatever reason. Thus, the primary analysis included 38 events in the suloctidil group and 47 in the placebo group (p = 0.17) representing a risk reduction of 24%. If total mortality is substituted for cardiovascular death, the corresponding figures are 47 in the suloctidil group and 58 in the placebo group (p = 0.08).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988160 TI - Mucosal damage simulating acute graft-versus-host reaction in cytomegalovirus colitis. PMID- 2988162 TI - [Genetic transformation of somatic cells. I. Clone of Chinese hamster cells defective in thymidine kinase and characterized by high transformation efficiency]. AB - A characteristics is given of clone A238 of the Chinese hamster cells deficient in thymidine kinase (TK). The isolation procedure is described. Upon transformation with the aid of DNA of plasmids, containing thymidine kinase gene (tk-gene) of Herpes simplex virus type 1 (HSV1) clone A238 cells show frequency (7.10(-5) and efficiency (130 TK+ colonies per 1 microgram of plasmid DNA) compatible with those of mouse line LMtk- cells. Modified transformation and selection conditions of clone A238 cells expressing TK-gene of HSV1 are demonstrated. A simple method is described for discriminating somatic cells, expressing either their proper or a virus TK-gene according to the cloning efficiency of cells on the HAT medium containing thymidine in concentration 100 micrograms/ml. It is shown that at the fixed total DNA concentrations a complete replacement of the eukaryotic carrier DNA for the plasmid DNA, containing no tg gene of HSV1, decreases but only insignificantly the frequency and efficiency of transformation. PMID- 2988163 TI - 11p13 deletion and reduced RBC catalase in a patient with aniridia, glaucoma and bilateral Wilms' tumor. AB - A rare case of a one-year-old child with Wilms' tumor, aniridia and glaucoma is described, in whom bone marrow chromosome analysis showed the presence of an interstitial microdeletion on the short arm of chromosome 11, presumably involving the p13 band. Research of the enzyme activity of RBC catalase showed a 40% reduction. This finding is compatible with the loss of the 11p13 band which contains the gene coding this enzyme. 11p13 deletion in Wilms' tumor and 13q interstitial deletion in retinoblastoma provide a rare case of prezygotic chromosome abnormality, which may be considered to have a determinant role in the tumor etiopathogenesis. PMID- 2988161 TI - [Partial suppression of rec B rec C mutations in E. coli by plasmid pBR 322 containing a Bacillus subtilis chromosome insertion]. AB - ATP-dependent DNAase genes of Bac. subtilis were originally cloned in E. coli plasmid pBR322. These genes are expressed in rec BC mutant E. coli cells leading to a complete recovery of the enzyme activity. Bac. subtilis enzyme suppresses reparative properties of the rec BC mutant to a considerable extent but is unable to replace functionally the E. coli mutant enzyme in recombination process. PMID- 2988164 TI - ABO blood-groups and the risk of gestational trophoblastic disease. AB - The relation between ABO blood group, mating patterns of patient/husband blood group, and the risk of gestational trophoblastic disease was investigated in a case-control study conducted in Milan on 286 women with histologically confirmed trophoblastic disease (245 benign hydatidiform moles and 41 persistent trophoblastic disease) and 433 control subjects admitted for normal delivery to the same hospitals where cases had been identified. ABO blood groups were associated with the risk of gestational trophoblastic disease (chi 2(6) for heterogeneity = 14.46, p = 0.02). Compared to women of group O or B, women of group A and AB had an elevated relative risk (RR) of benign mole (RR = 1.4 and 2.3, respectively). The risk estimates were higher for persistent trophoblastic disease, i.e., 2.2 for women of group A and 4.8 of group AB. The tests for linear trend in risk from benign to persistent disease were statistically significant in both A and AB groups. There was a significant interaction between blood group and age, since the ABO-related risk was elevated only for women over the age of 35. When mating combinations of maternal/paternal blood groups were considered, women of group A married to males of group O had a risk estimate not substantially different than those married to group A males. PMID- 2988165 TI - Inflammatory malignant fibrous histiocytoma of the kidney: an immunohistochemical and ultrastructural study. AB - A primary inflammatory malignant fibrous histiocytoma of the kidney is reported. The differential diagnosis from other sarcomas and pseudosarcomatous lesions is discussed on the grounds of the histologic, immunohistochemical and ultrastructural results. The neoplasm was made up of histiocytes, fibroblasts, myofibroblasts, foam cells and undifferentiated mesenchymal cells with admixed granulocytes, lymphocytes and plasma cells. The immunohistochemical study showed a positivity for alpha-1-antichymotrypsin and a weak positivity for alpha-1 antitrypsin in mononuclear and pleomorphic multinucleated tumor cells. PAS positive, diastase-resistant intracytoplasmic hyaline globules in necrotic cells, examined by electron microscopy, most likely represent lysosomal structures, in accordance with the theories of De Duve, Vattiaux and Von Ardenne. PMID- 2988166 TI - Comparison of the chemotactic response to conditioned medium of BALB/c3T3 fibroblasts and their SV 40 transformants. AB - The chemotactic migration of transformed cells out of their tissue of origin might represent an essential component of tumor invasion and metastasis. In the Boyden chamber assay SV 40 virus-transformed BALB-c3T3 fibroblast (SV/3T3) showed an increased chemotactic response to fibroblasts-conditioned medium in comparison to non-transformed BALB/c3T3 cells. This was verified for a large range of incubation times and cell densities. Fibroblast-conditioned medium contains components of the extracellular matrix of connective tissue. Since organs rich in connective tissue are often the site of metastasis, the enhanced chemotaxis of virus-transformed cells to conditioned medium could contribute to explain the malignancy of these cells. PMID- 2988167 TI - [Biochemical mechanisms of fluorine action]. AB - Biochemical mechanisms of fluorine ion action accounting for the biological role and significance of fluorine for vital activity of the organism are investigated. Results of these investigations are generalized. This trace element is shown to participate at least in two vitally important systems of the organism: the adenylate cyclase system which accounts for the cell response to neuroendocrinological information and the immune protection system providing antimicrobic resistance. Available data permit considering that cytotoxic fluorine action is based on the ability to hinder protein synthesis in eukaryotes and to stimulate peroxidation processes of biomembrane lipids. Inorganic fluorine compounds are recommended to be used with the treatment-and-prophylactic purpose for certain pathologic states, associated with its insufficient or excessive arrival into the organism. PMID- 2988168 TI - [The effect of ATP analogues on ATPase and phosphatase activities of Na+, K+ ATPase for duck salt glands]. AB - ATP analogues are studied for their effect on phosphatase and ATPase activities of Na+, K+-ATPase with the aim to obtain data concerning properties and structure of sites of high and low affinity to ATP. The activating effect of nucleotides on K+-dependent phosphatase reflecting their ability to be bound with the centres of high affinity decreases in a series: ATP, N1-oxy-ATP, CTP, JTP. In the domain of high ATP concentrations, where low affinity site is saturated, ADP is a competitive inhibition of ATPase reaction with Ki of 300 microM. The analysis of N1-oxy-ATP inhibiting effect has shown that its affinity to this site is six times less than that of ADP. The absence of the inhibiting effect of CDP, JDP, GDP and UDP in concentrations up to 10 mM testifies to the fact that sites of both low and high affinity to ATP are characterized by high specificity with respect to the adenine part of the substrate molecule. PMID- 2988169 TI - [The influence of diphosphonic analogues of inorganic pyrophosphate on activity of RNA-polymerases from the calf thymus]. AB - Diphosphonic analogues of inorganic pyrophosphate (PPi): methylene-, oxyethylidene-, aminomethylenediphosphonic acids as well as phosphonacetic, imidodiphosphoric bis- (phosphonomethyl)-phosphonic acids and methylenediphosphonic and phosphonic acid monoanhydrides were studied for their effect on the RNA-synthesizing activity of thymocytes. DNA-dependent RNA polymerases I and II from the calf thymus nuclei were used for these studies. The analogues and PPi under study are shown to be inhibitors of both RNA-polymerases in nuclei from calf thymus and of purified RNA-polymerase II, which is more sensitive to the effect of diphosphonates. Methylenediphosphonic acid is the strongest inhibitor among the studied analogues, and imidodiphosphoric and phosphonacetic acids are the weakest inhibitors. Inhibition of purified RNA polymerase II by diphosphonates has a complex character and includes both interaction of the PPi analogues with enzymes and chelating by them of Mn ions which are cofactors for RNA polymerase. PMID- 2988170 TI - [The influence of diphosphonic analogues of inorganic pyrophosphate on reactions catalyzed by DNA-dependent RNA-polymerase II]. AB - Diphosphonic analogues of inorganic pyrophosphate were studied for their influence both on RNA pyrophosphorolysis and pyrophosphate exchange, catalyzed by purified DNA-dependent RNA-polymerase II from calf thymus. Methylene-, oxyethylene-and aminomethylenediphosphonic acids are shown to compete with PPi for incorporation into nucleoside triphosphate. They activate RNA pyrophosphorolysis in the concentration of 2 mM, but to a less extent than PPi does. PMID- 2988171 TI - [Activity of guanosine diphosphatase and guanosine triphosphatase in the rat brain and liver under radiation injury]. AB - The activity of guanosine diphosphatase (GDPase) and guanosine triphosphatase (GTPase) was determined in mitochondrial fractions and postmitochondrial supernatant fluid from the rat brain and liver nuclei 1, 3, 6, 24 and 48 hours after their irradiation by gamma-quanta of [60Co] in a dose of 774 mC/kg. Under the influence of irradiation the activity of GDPase in the liver and brain mitochondria increased, that of GTPase in the liver mitochondria underwent phase changes. The GDPase activity tended to increase in postmitochondrial supernatant fraction of the liver and brain; the GTPase activity was at first inhibited and then increased. PMID- 2988172 TI - [Effect of tuftsin, a neuropeptide and its derivatives on the content of nicotinamide coenzymes and the activity of cytochrome- c-oxidase of brain tissue]. AB - Tuftsin, a neuropeptide, and its derivatives are studied for their influence on the content of oxidized and reduced forms of nicotinamide enzymes as well as on the activity of cytochrome-c-oxidase of the cortex and limbic system. Peptides under study are shown to increase the content of nicotinamide coenzymes, mainly due to the oxidized forms of NAD and enhance the activity of cytochrome-c-oxidase as well mainly in the emotiogenic brain structures. Under these conditions tuftsin and its derivatives intensify cellular respiration of the neurons. PMID- 2988174 TI - [Synovial sarcoma of the larynx]. PMID- 2988173 TI - [Differential diagnosis of juvenile angiofibroma of the base of the skull]. PMID- 2988175 TI - [Clinical significance of pulsatory noise in the ear]. PMID- 2988176 TI - [Blood circulation in the fingers and electric activity of the muscles after the surgical treatment of children with ectrodactylia]. AB - The rheographic examination of hands in patients aged from 2 to 15 with ectrosyndactyly has revealed a less volume of blood flow, worse elastic properties of the vessels of diseased fingers and lower level of the electric activity of interosseous muscles of the hand. Elimination of syndactyly with the help of different variants of operations did not cause complications in the postoperative period associated with the disturbed blood flow in the fingers. PMID- 2988177 TI - Intimal asteroid bodies in horses: light and electron microscopic observations. AB - The morphology of asteroid bodies in equine arteries is demonstrated by light microscopy as well as by scanning and transmission electron microscopy combined with chemical analysis. Asteroid bodies first occur in horses at four weeks of age in all investigated tissues except the esophagus and always are located in the subendothelial space. The number, shape, ultrastructure and chemical composition of asteroid bodies differ markedly--depending on the age of the horse. Asteroid bodies are round and smooth in foals, but are shaped irregularly and have several projections and marked stratified calcification in adult horses. Asteroid bodies probably originate from smooth muscle cells. No direct etiological connection between asteroid bodies and migrating Strongylus vulgaris could be verified. PMID- 2988179 TI - Suspected hydrocyanic acid poisoning in cattle. PMID- 2988178 TI - Exacerbation of murine respiratory mycoplasmosis in gnotobiotic F344/N rats by Sendai virus infection. AB - Six- to eight-week-old gnotobiotic F344/N rats were inoculated intranasally with 10(5.0) colony-forming units of Mycoplasma pulmonis or were sham inoculated, then one week later were given 10(0.2) 50% tissue culture infective doses of Sendai virus or sterile medium. Groups of rats were killed immediately after virus inoculation and three, five, ten, and 20 days later. Lesions in nasal passages, middle ears, larynxes, tracheas, and lungs from half of the rats in each group were subjectively scored. Organs from the other rats were quantitatively cultured for M. pulmonis and for Sendai virus. Rats given Sendai virus alone had mild, patchy, necrotizing rhinitis, laryngitis, tracheitis, and bronchitis, but not bronchiolitis or interstitial pneumonia. M. pulmonis alone induced mild lesions of murine respiratory mycoplasmosis including mild to moderate suppurative rhinitis, otitis media, laryngitis, and tracheitis with submucosal lymphoid accumulation and epithelial hyperplasia, but not lung lesions. Rats given M. pulmonis and Sendai virus had severe lesions characteristic of advanced mycoplasmal disease throughout the respiratory tract, including suppurative bronchitis with extensive lymphoid accumulations and epithelial hyperplasia; some rats also had suppurative pneumonia and bronchiectasis. Larger numbers of M. pulmonis colony-forming units were in rats given Sendai virus, but there was no statistically significant difference in Sendai virus infectious units between rats also given M. pulmonis and those given virus only. PMID- 2988180 TI - Isolation of avian infectious bronchitis viruses in Morocco including an enterotropic variant. PMID- 2988181 TI - Isolation and identification of bovid enteroviruses from free-living wild animals in Botswana. PMID- 2988182 TI - Serum antibody to respiratory syncytial virus in goats in the UK. PMID- 2988183 TI - Rumen papillomas in sheep. AB - Out of a sample of 200 rumens from sheep slaughtered in Edinburgh, papillomas were found in 25. They occurred as fibro-papillomas, mostly along the pillar of muscle between the dorsal and ventral rumen, and were often multiple. No inclusion bodies were seen in the lesions or papilloma virus particles on electron microscopy. Homogenisation of papillomatous tissue followed by various methods of purification did not yield identifiable virus particles, and viral DNA was not detected. Immunoperoxidase staining showed a very small number of positive cells at or on the surface of 6 out of 10 lesions examined. Thus, it seems probable that virus particles are not found in large numbers in the rumen papillomas of sheep, unlike the situation in ovine skin warts, but are present in a few epithelial cells which are rapidly shed from the surface of the mucous membrane. PMID- 2988184 TI - An attempt to protect calves against experimental bovine leukosis using allografts. AB - Six calves sensitised by implanting skin from a calf were later inoculated with lymphocytes from the same calf after the calf had been infected with bovine leukosis virus (BLV). Two out of 6 calves challenged did not develop BLV antibodies and BLV was not isolated from these animals, whereas all of the 5 control calves became infected with BLV. PMID- 2988185 TI - Inability to detect a K cell in bovine peripheral blood leukocytes. AB - Antibody-dependent cellular cytotoxicity to viral-infected cells, chicken red blood cells, and tumor cells was tested using different effector cell populations: polymorphonuclear cells, mononuclear cells, and mononuclear cells separated into adherent and nonadherent populations by Sephadex G-10. Polymorphonuclear cells were the most efficient mediators of antibody-dependent cellular cytotoxicity against most targets, although a combination of G-10 adherent and polymorphonuclear cells was more efficient in killing infectious bovine rhinotracheitis virus-infected cells than either single cell population. Removal of G-10 adherent cells from the mononuclear cell population removed all antibody-dependent cellular cytotoxicity from that population, indicating the lack of a typical K cell in bovine peripheral blood. PMID- 2988186 TI - Isolation of visna-maedi virus from the choroid plexus of an apparently healthy sheep in Italy. AB - In this paper, the presence of visna-maedi in Italy, reported to exist since 1982 on clinical grounds, and the presence of anti-visna-maedi antibody is confirmed by the isolation of the viral agent from choroid plexus cells of an apparently healthy sheep. Gel diffusion test antigen, prepared from the explanted choroid plexus cells and from normal choroid plexus cells infected with the isolated agent, gave a positive reaction with one or two identity lines with reference antigens. The isolated agent gave both a characteristic cytopathic effect and a cytoplasmic fluorescence starting 24 hours after infection in normal choroid plexus cell cultures. Fluorescence was observed neither in control normal choroid plexus cells nor in infected normal choroid plexus cells incubated with a visna maedi negative serum. By electron microscopy, budding forms arising from the cell membrane and extracellular particles of two distinct types were observed. One type was characterized by an electron-dense central core and a single membrane, and ranged from 80 to 110 nm, while the other had elongated bar-like cores and a double wall, and ranged from 100 to 140 nm. The characteristics observed for the isolated agent are identical to those reported by various authors for visna-maedi virus. PMID- 2988187 TI - Advanced gastric cancer and prognosis. AB - The morphological features of 158 gastric carcinomas were analyzed in an attempt to identify patterns best correlated with prognosis. To this end, the depth of infiltration, vascular invasion, intra- and perineoplastic lymphocytic infiltrate, lymph node metastases and number of metastatic lymph nodes were evaluated according to the several classifications for advanced gastric cancer. A good correlation between prognosis and histological features of malignancy were observed, as well as different five-year survival rates for Mulligan, Lauren and Ming histotypes. However, when the influence of each single morphological criteria of malignancy was examined, these differences disappeared for Mulligan and Lauren histotypes. On the other hand, the better prognosis for Ming expanding type carcinomas appeared unrelated to any individual feature of malignancy. PMID- 2988188 TI - Epithelial markers for paraffin-embedded human tissues. Immunohistochemistry with monoclonal antibodies against milk fat globule antigens. AB - About 200 human tumours and corresponding normal tissue samples were investigated by immunoperoxidase tests for the expression of MAM-3, MAM-5 and MAM-6 antigens, which had previously been defined by monoclonal antibodies to human milk fat globule membranes. All tissue specimens had been treated for routine histopathology, i.e. fixed in formalin and embedded in paraffin. One of the antigens, MAM-6, appeared to be an important epithelial marker, present in all normal and neoplastic breast tissue samples, in about 80% of non-mammary normal tissues and in more than 90% of non-mammary epithelial tumours. It could never be detected in normal and neoplastic mesenchymal and neuroectodermal structures. Direct comparison with the distribution of Carcinoembryonic Antigen (CEA), Tissue Polypeptide Antigen (TPA) and keratin provided clear evidence that MAM-6 is different from these well known epithelial markers. MAM-3 proved to be an additional important marker exhibiting a characteristic distribution pattern in those epithelial tissues investigated. In contrast to MAM-6, it could never be detected in renal cell cancers and carcinomas of the prostate gland, thus allowing differential diagnosis on the basis of immunohistochemistry. MAM-5, known to be associated with lactoferrin, was mainly detectable in secretory organs and their tumours. In the group of breast tumours, its expression was mainly seen in lobular cancers. These findings suggest a use for these new markers for routine histopathology. PMID- 2988189 TI - Immunohistochemical localization of HCG and its subunits in testicular germ cell tumours. AB - Native hCG and its alpha and beta subunits have been localized by immunocytochemistry in 45 testicular germ cell tumours of the testis. Positivity was found for the three molecules in areas with trophoblastic differentiation or in syncytial-like giant cells present in some seminomas. Isolated positivity of hCG alpha was demonstrated in isolated cells usually found in areas of entodermal differentiation of immature malignant teratomas, and probably of neuro-endocrine function. This finding points to genomic derepression in tumour cells and probably also indicates a variability in subunit synthesis and a defect in subunit recombination. PMID- 2988190 TI - Epidemiological features of influenza in a large town of Romania during 1983. AB - Data supplied by the active influenza surveillance - including clinical, epidemiological and laboratory investigations - allowed the characterization of the particularities of an influenza outbreak caused by A(H3N2) virus in a large town of Romania in March 1983. The epidemiological impact of influenza in 1983 is discussed in the light of the complex relationships between the antigenic structure of circulating influenza strains and the profile of antiinfluenza immunity of the population. PMID- 2988191 TI - Virus - tumor cell relationships. In vivo cocultivation of para-influenza type 1 (Sendai) virus and of Rous sarcoma virus (Schmidt-Ruppin strain) in mouse Ehrlich ascites carcinoma. AB - Co-infection of Ehrlich ascites carcinoma (EAC)-bearing mice with Sendai virus and Rous sarcoma virus (RSV) did not result in the formation of complete RSV. Sendai virus could be, however, propagated in this system over 8 serial passages. As demonstrated by immunofluorescence and complement fixation reactions, antigens specific to each virus were synthesized in EAC cells following either single or mixed virus infection. The virus progens also contained antigenic fractions incorporated from the host cell. The incomplete progens synthesized when RSV inoculation preceded that of Sendai virus possessed three polypeptide fractions characteristic of Sendai virus and one RSV-specific fraction. PMID- 2988192 TI - Detection by immunofluorescence of viral, chlamydial and mycoplasma antigens in men with urethritis. AB - The presence of viral, chlamydial and mycoplasma antigens was investigated by the indirect immunofluorescence (IF) reaction in the urethral cells of 30 male patients with urethritis. Positive IF reactions were recorded in 21 (70%) of the cases, 2-6 different antigens being simultaneously visualized in 14 patients. In addition to chlamydial, mycoplasma and herpes antigens, parainfluenza virus and adenovirus antigens were visualized in 39.34% and 11.47% of the cases, respectively. The efficacy of the IF technique in the rapid laboratory diagnosis of male urethritis is discussed. PMID- 2988193 TI - Effect of homologous tumor DNA on the evolution of SV40-induced hamster sarcoma. AB - DNA extracted from SV40-induced hamster sarcoma (SV40--HS DNA) increased the survival length of animals carrying the homologous tumor and--in some cases- inhibited the development of the tumor. The efficacy of the preparation was directly proportional to the number of administrations; it did not necessarily depend on the amount of SV40--HS DNA per dose. SV40 DNA had no favourable effect on the evolution of the SV40-induced tumor, which suggests that viral DNA does not represent the active component of the SV40--HS DNA preparations. Some possible mechanisms of the effect of homologous tumor preparations are discussed. PMID- 2988194 TI - [Effect of immunosuppressants on the enhanced sensitivity of white mice to the eastern equine encephalitis virus]. AB - EEE virus infection could be induced in white mice whose natural resistance to this virus had been reduced by immunosuppressive drugs such as: Adreson, Cyclophosphan and hydrocortisone. PMID- 2988196 TI - Plaque morphology is altered by the label of inorganic components of the cell culture medium. PMID- 2988195 TI - Some trends of research in the domain of viral neuroinfections approached in the "Stefan S. Nicolau" Institute of Virology. AB - The main directions of research in the field of viral neuroinfections approached during 35 years in the Institute of Virology are briefly outlined. After some considerations on terminology and on the classification of viral encephalitides, mention is made of the studies in the domain of herpes infections, rabies, meningitis, encephalitis and slow virus infections of the central nervous system (subacute sclerosing panencephalitis, multiple sclerosis, amyotrophic lateral sclerosis, Creutzfeldt-Jakob disease, etc.). PMID- 2988197 TI - Influence of some organic additives to the culture medium on the size of poliovirus-induced plaques. PMID- 2988198 TI - [Analysis of hospital morbidity from adenovirus keratoconjunctivitis]. PMID- 2988199 TI - [Use of microcomputers for the topical diagnosis of injuries and diseases of the peripheral nervous system]. PMID- 2988200 TI - [Coupling of insulin-stimulated ATP formation by preparations of plasma membrane enriched particles with proton transport across the plasma membranes of target cells]. AB - In the particles enriched with plasmatic membranes of target cells (human erythrocytes, skeletal muscles and adipocytes of rats) ATP was steadily formed within 1 min of incubation of the particles with insulin (4 microgram/ml) in the medium containing Tris-HCl buffer, pH 7.5, ADP, Mg2+, inorganic phosphate, NaF, during NADH oxidation in presence of cytochrome c and oxygen (30 degrees). Introduction of proton ionophore p-trifluoromethoxycarbonyl cyanide phenylhydrazone (pFCCP) into the incubation medium before addition of human erythrocyte particles enriched with plasmatic membranes and of insulin led to a distinct increase of the insulin-stimulated accumulation of ATP in erythrocyte plasmatic membranes: at 10(-6) M concentration of pFCCP the increase was 3.5 fold, at 5 X 10(-6) M of pFCCP it was even 8-fold. In presence of 10(-4) M pFCCP the insulin-stimulated ATP accumulation was increased 2-fold in the particles from rat adipocytes. Output of protons into the medium and its acidification (alteration of pM by 0.01-0.012) within 3-6 sec was found in experiments with impulse administration of insulin 100 micrograms and pFCCP 0.1 micrograms into the suspension of freshly isolated rat adipocytes (5 ml) added to an electrolyte containing 150 mM NaCl, 10 mM beta-hydroxybutyric acid, 0.01 mM Gly-Gly (pH 7.45). PMID- 2988201 TI - [Restriction endonucleases from Shigella sonnei 47]. AB - Two restrictases SsoI and SsoII, belonging to the enzymes of restriction of the class II, were isolated from a strain of dysenteric bacteria. The structure of the site sensitive to SsoI and SsoII was studied after fragmentation of testor DNA as well as by means of direct determination of nucleotide sequence. SsoI was shown to be an isoschizomer of the EcoRI restrictase from E. coli. Restrictase SsoII proved to be a new enzyme, which hydrolyzed the sequence 5' ...CCNGG.. 3' and was distinct from the known restrictases as shown by studies of the type of DNA hydrolyzed. A three-step procedure is developed for isolation of SsoII restrictase involving the consecutive chromatography on Blue Sepharose, phosphocellulose PII and phenyl-Sepharose. Restrictase SsoI and EcoRI were isolated by means of isoelectrofocusing using ampholines. PMID- 2988202 TI - [Cyclic nucleotide content of the rabbit myometrium in various functional states]. AB - Myometrium of female rabbits at the state of functional rest contained 260 pmoles of cAMP per g of tissue and 25 pmoles cGMP. In dynamics of pregnancy content of cAMP was increased up to 400 pmoles/g within the second half of pregnancy; the content of cGMP was decreased down to 10 pmoles/g. During labor content of cAMP and of cGMP became minimal; within the postnatal period concentration of cAMP was increased up to the values observed in the antenatal period and the content of cGMP was unaltered. Under these conditions the content of cAMP, cGMP and the activity of corresponding soluble phosphodiesterases were similarly altered. The regulatory subunit of protein kinase of the type I (RI) was shown to possess two sites for cAMP binding and RII--one site. The data obtained suggest that cyclic nucleotides are important for regulation of the myometrium functional state. PMID- 2988203 TI - [Lung apudomas]. AB - As a result of a complex morphological examination of 140 cases of lung neoplasms, 24 apudomas (17.11%) and 3 carcinomas (2.14%) were identified. The latter consisted of histologically-heterogeneous lesions containing endocrine cells in combination with epidermoid or glandular cells. Clinical symptoms as well as cytologic, histological, histochemical and electron microscopic characteristics of the tumors were studied. Most of those tumors were poorly differentiated, histologically identical to various types of small-cell lung carcinoma, or--less frequently--solid lesions and other types of tumor. The above tumors can be detected by special diagnostic procedures. PMID- 2988206 TI - [Monkeys as a source of human viral diseases]. PMID- 2988205 TI - [Effect of the type of nutrition on cyclic nucleotide metabolism in workers in the phosphate industry]. PMID- 2988204 TI - [T-activin in the combined treatment of patients with small cell cancer of the lung]. AB - T-activin immunotherapy in conjunction with maintenance chemotherapy was given to 13 patients suffering small-cell carcinoma of the lung in whom previous cytostatic and combined treatment (chemotherapy+radiation) resulted in a complete regression of tumor. The median survival time in those patients was 113 weeks versus 75 weeks in 13 controls. The results make the case for further studies on application of the said agent aimed at longer survival and remission in patients with small-cell carcinoma of the lung. PMID- 2988208 TI - [Identification by electron microscopy of a corona-like virus inducing diarrhea in piglets]. AB - The results of identification by means of electron microscopy and immune electron microscopy of a corona-like virus isolated in diarrhea of pigs during the first weeks of life are presented. Morphological forms, diameter of particles, the length of external processes, and the buoyant density of virions were determined. PMID- 2988207 TI - [Reproduction of the vesicular stomatitis virus in mouse splenocytes]. AB - Interaction of vesicular stomatitis virus with immunocompetent cells in vivo and in vitro was studied. In intact mice, a prolonged infection of the spleens with low levels of virus replication was observed. Experiments with various splenocyte cultures demonstrated differences in their capacity to support virus reproduction. A relationship between the functional activity of the cells (the level of spontaneous blastogenesis, index of PHA response) and levels of virus reproduction was shown. Mitogen stimulation of blastogenesis was accompanied by intensified replication in reactive cultures. In splenocyte cultures from immune animals, reproduction levels were decreased and virus replication stopped. PMID- 2988209 TI - [Model of experimental Pixuna infection in white mice]. AB - The results of the development of a nonfatal model of experimental infection induced by Pixuna alpha-virus and suitable for the evaluation of the effectiveness of antiviral drugs are described. The asymptomatic infection induced in white mice by intranasal inoculation of Pixuna virus is characterized by intensive virus multiplication in the brain and spleen of the animals. In these organs virus reproduction is observed early after infection and amplification of the agent reaches maximum titres within 72 h postinfection. With this model, the main criterion of the effectiveness of antiviral drugs would be their effect on virus reproduction in the spleen and brain. PMID- 2988212 TI - [Diagnosis of rotavirus infection using coagglutination reactions]. PMID- 2988211 TI - [Phenotypic mixing of the Rous sarcoma virus with the Sendai virus and the biological properties of the phenotypically mixed virions]. PMID- 2988210 TI - [Epidemiological and immunological research on cytomegaly in parturients and newborn infants among the indigenous and immigrant population in the Far North]. AB - In the immigrant population of the Central Yakutiya, cytomegalic cells in the saliva and urine are found much more frequently than in the indigenous population (P less than 0.001). Significant differences were found in the content of complement-fixing antibody to CMV in parturients and newborns in the Extreme North as compared with the European USSR. While in the latter complement-fixing antibodies to CMV are more frequently found in newborn babies (86% in newborns and 69.8% in parturients), in the Extreme North it is vice versa (52.53% in parturients and 32.73% in newborns). Large numbers of seronegative women of the indigenous and immigrant population becoming pregnant get into the group of risk of infection with CMV which is explained by a high rate of detection of IgM in newborn babies of the indigenous (8.69%) and immigrant (9.8%) population and indicates congenital CMV infection. PMID- 2988214 TI - [Development and use of an immunoenzyme method for detecting interferon]. PMID- 2988213 TI - [Use of human embryonic fibroblasts for the laboratory examination of serious meningitis patients]. PMID- 2988215 TI - [Isolation of somatic cell hybrids by using a specific interferon- virus method of selection]. PMID- 2988216 TI - [Cytomegalovirus infection in a newborn infant primarily diagnosed as hemolytic disease of newborn]. PMID- 2988217 TI - [Acquired immunodeficiency syndrome]. PMID- 2988218 TI - Effect of Acarbose on the production of hydrogen and methane and on hormonal parameters in young adults under standardized low-fibre mixed diets. AB - Short and middle term effects of Acarbose were studied in volunteers on a standardized, low-fibre, mixed diet for the development of tolerance phenomena with gas exhalations and some peptide hormone levels as main parameters. Both hydrogen and methane were measured quantitatively as diurnal profiles. Acarbose caused an about 20-fold increase of H2 exhalation and had only moderate effects on methane production, indicating the presence of fermentable carbohydrates in the large bowel. Methanogenic individuals exhaled significantly less H2 than did non-methanogenic subjects. Changes in blood glucose, serum insulin, GIP, gastrin, and plasma glucagon, caused by Acarbose, reflected delayed glucose absorption and were plausible within the regulatory framework of carbohydrate assimilation. When the Acarbose regime was maintained for 5 weeks on a controlled diet, abdominal sensations like e.g. meteorism declined remarkably while carbohydrate fermentation remained high and lowered GIP was sustained. Thus functional responses of the gastro-intestinal tract to altered carbohydrate supplies, elicited by Acarbose, were found by 3 independent parameters: anaerobic gas production, peptide hormone levels, and subjective abdominal sensations. The objective parameters seem to remain constant in the longer run, while subjective parameters show long-term adaptation. PMID- 2988220 TI - Fluoride content of selected human food, pet food and related materials. AB - A survey was made of the fluoride content of selected human foods, some animal feeds, and related materials, with the purpose of a better characterization of products whose fluoride may contribute to the intake of fluoride by man as well as by some kinds of animals. Uptake of fluoride, especially from food with or without a foregoing food chain, was found to be more widely spread than has been documented previously. The major source of fluoride is represented by marine organisms, regardless of the ways by which man or animals are exposed to fluoride. As a consequence of the detailed analytical data given, recommendations would extend towards i) a more wide-spread analytical control of foods and feeds for fluoride, ii) an intensified assessment of fluoride bioavailabilities from foods and feeds, and iii) the consideration of such data in the evaluation of fluoride supplementations for optimized intakes, as recommended. PMID- 2988219 TI - [Vitamin D sterol in human milk, cow's milk and baby food]. AB - A review is given of the results of vitamin D determinations in human and cow's milk using physico-chemical methods. Thereby only parent vitamin D is determined. Further, the results obtained with protein-binding assays are discussed in detail. With the aid of these newer methods the concentrations of hydroxylated vitamin D derivatives can also be measured. A comparison of the concentration of these vitamin D metabolites in human milk and infant formulas based upon cow's milk suggests that the higher anti-rachitic activity of human milk is connected with its higher concentration of 25-hydroxy-vitamin D. PMID- 2988221 TI - [Biochemistry and significance of prostaglandins, prostacyclins, thromboxanes and leukotrienes and the modification of their biosynthesis by essential fatty acids]. AB - A survey is given of the biochemistry and importance of the prostaglandins, the thromboxanes, the prostacyclins and the leukotrienes. The formation of these compounds takes place from poly-unsaturated fatty acids and is regulated: here the phospholipase A2 plays a role. Among others the prostaglandins take part in the regulation of the blood supply of various tissues (heart, kidneys), in the regulation of the reproduction and in the evokation of labour. The thromboxanes and prostacyclins influence the aggregation and the desaggregation of the thrombocytes, respectively. The leukotrienes take part in the regulation of the permeability of the capillaries, in the migration of the leucocytes, the formation of inflammatory processes and in the evokation of asthma bronchiale. The insufficient intake leads to functional disturbances in various tissues, which partly are to be traced back to a decrease of the formation of the tissue hormones mentioned. An intake transgressing the demand has a favourable effect in various diseases: thus the inclination to aggregation of the thrombocytes is decreased and the development of arteriosclerotic changes is inhibited. PMID- 2988222 TI - [Eccrine porocarcinoma]. AB - We report on a 74-year-old female suffering from an eccrine porocarcinoma nea the mons pubis. Two additional tumors appeared in the same area. The three tumors revealed different stages of development. PMID- 2988223 TI - [Fiber-rich snacks with reference to their effect on the digestive activity and blood lipids of the elderly]. AB - 32 subjects of a home for the elderly and nursing home were fed special test meals beside the usual home diet for 84 test days. The test meals, rich in dietary fibre, "Leinsamen" (linseed) and "Rohfasergemisch" (dietary fibre mix) are adequate for improving the subjects' digestion. The identified reductions are insignificant in regard to the effect on an increased serum triglyceride level. With the snack "Fruchtemuesli" we achieved a highly significant, with "Leinsamen" a significant reduction of blood cholesterol level. PMID- 2988225 TI - Prospects for improved foot and mouth disease vaccines. AB - Improved foot and mouth disease (FMD) vaccines which have increased efficiency and stability are highly desirable. Work which aims to increase the stability of FMD virus particles for vaccine use is described and the use of these particles and of antigenic fragments of the virus for controlled release vaccine products is considered. PMID- 2988224 TI - Nucleotide sequence of bovine 1.723 satellite DNA. AB - The nucleotide sequence of the bovine 1.723 satellite DNA repeated unit was determined. The 680 bp long period of this satellite DNA does not show any significant sequence similarities with the known bovine satellite DNAs. Short repetitive sequences which are parts of 680 bp long repeated units do not form any orderly periodical structure. It seems, however, that the basic repeated unit of the 1.723 bovine satellite DNA has been formed by successive duplications of two, about 100 bp long sequences. The sequence divergence between different copies of the 680 bp repeated unit was also analyzed. PMID- 2988226 TI - Novel approaches to foot and mouth disease vaccination. AB - In view of the problems associated with the use of conventional foot and mouth disease (FMD) vaccines, such as antigenic variability of the FMD virus, the cost of vaccination programmes and problems associated with vaccine storage, alternative approaches to the design of FMD virus are discussed. Identification of the antigenic site of the FMD virus and isolation or synthesis of the peptides which make up the antigenic site are considered for vaccine use. PMID- 2988227 TI - Bluetongue vaccine: cells and/or antibodies. AB - Immunological studies with bluetongue virus have indicated that protection from re-infection involves components of both the humoral and cellular immune response. However, it was found that the humoral response was type-specific, whilst the cellular immune response, particularly through the action of cross reactive cytotoxic T lymphocytes, gave rise to heterotypic protection. Work involving simultaneous and sequential inoculation of live virus and the inoculation of various inactivated preparations has further characterized the type of vaccine formulation needed for efficient protection in multitype endemic areas. The authors cite these studies on bluetongue virus as an example of an immunological approach to vaccine design that is too often ignored by vaccine manufacturers and yet clearly yields results. PMID- 2988228 TI - Evidence for the existence of a thermolabile element in adenovirus type 5 that plays a role during an early stage of infection. AB - In an attempt to learn the molecular mechanism behind the initial steps (before genome expression) of adenovirus infection, the effect of temperature treatment of adenovirus type 5 particles on their infectivity was studied. It was found that adenovirus type 5 could be inactivated by temperature treatment at between 43 and 44 degrees C. Both biochemical and biophysical examination of adenovirus particles failed to disclose any significant changes after exposure to temperatures up to 45 degrees C. Further, viral particles treated with the same temperature range retained their ability to attach to cell surface receptors and to penetrate the plasma membrane. Application of a temperature pulse of 44 degrees C to adenovirus-infected HeLa cells indicates that the infection can be inhibited by exposure of infected cells to a brief temperature pulse of 44 degrees C between 10 and 50 min postadsorption with a mid-point of 30 min postadsorption. In addition, electron microscopic examination of HeLa cells following a temperature pulse of 44 degrees C suggests that there is a failure to accumulate viral particles at the perinuclear region. These findings support the hypothesis that adenovirus contains a thermolabile element which plays a crucial role during an early stage of infection. Possible correlation of this element with the adenovirus endogenous protein kinase is also examined and discussed. PMID- 2988229 TI - Mammary tumorigenesis in C3Hf/Ki mice: examination of germinal mouse mammary tumor viruses and the int-1 and int-2 putative proto-oncogenes. AB - The organization and expression of endogenous mouse mammary tumor virus (MMTV) proviruses in normal and neoplastic C3Hf/Ki tissues were examined. MMTV containing EcoRI, HindIII, BamHI and PstI restriction fragments of C3Hf/Ki DNA were identical to those of C3H/StWi DNA. The full-length endogenous MMTV Units Ia (Mtv-7), II (Mtv-8), III (Mtv-9) and IV (Mtv-10), in addition to the subgenomic endogenous MMTV Units I (Mtv-6) and IX (Mtv-14), were germinally transmitted in C3Hf/Ki DNA. The previously uncharacterized Mtv-7 was contained in EcoRI fragments of 16.7 and 11.7 kbp. The endogenous MMTV Unit V (Mtv-1), which is responsible for virus production and mammary tumorigenesis in C3Hf/He mice, was absent from C3Hf/Ki DNA. The 9.0 kb gag-pol, the 3.8 kb env and the 1.7 kb LTR MMTV RNA transcripts were present in C3Hf/Ki mammary glands. MMTV proviruses, in addition to the endogenous C3Hf/Ki MMTV complement, were not detected in C3Hf/Ki mammary tumor DNA. The DNA organization and RNA expression of the putative mammary proto-oncogene regions int-1 and int-2 were also examined in C3Hf/Ki mammary tumors. The int-1 and int-2 regions did not appear rearranged, amplified, or expressed in C3Hf/Ki mammary tumors. These studies indicate that MMTV proviral activation of the int proto-oncogenes is not necessary for C3Hf/Ki mammary tumorigenesis. PMID- 2988232 TI - The diagnostic value of perinatal ultrasound monitoring for foetal abdominal mass. AB - In the past two years seven cases of foetal abdominal mass were observed prenatally and operated on within 30 days of life. In four of seven cases no abdominal mass was ever palpable neither at birth nor under anaesthesia just before laparotomy, whereas in all cases it was both sonographically and surgically detected. The authors stress the importance of perinatal ultrasound monitoring to cope early with clinically undetectable abdominal mass. Furthermore, they emphasise that the necessary condition for definition of a mass should by now be that it is palpable and seen via ultrasound, but a perinatal ultrasound image of mass is sufficient to warrant careful detailed clinical evaluation. PMID- 2988230 TI - Immunologic characterization of herpes simplex virus type 2 antigens ICP10 and ICSP11/12. AB - Infected cell protein 10 (ICP10) or antigen 4 (Ag4) and infected cell-specific protein 11/12 (ICSP11/12) have been suggested as specific antigenic markers for cervical carcinoma. Experiments were designed to determine whether ICP10 and ICSP11/12 are distinct antigens and to determine the cellular localization of ICP10. Results indicate that an apparent 160 kdalton (kDa) protein analyzed by 8.5% polyacrylamide gels (= 144 kDa protein analyzed by 7.0% polyacrylamide gels) was detected in HSV-2-infected but not mock-infected extracts. This protein is an early virus-induced protein appearing 2-4 h after HSV-2 infection, and it was synthesized in the presence of successive blocks with cycloheximide and actinomycin D. These properties are characteristic for ICP10 (Ag4), thus establishing the identity of the 160 kDa/144 kDa protein as ICP10. Furthermore, Western blot analyses indicated that ICP10 and ICSP11/12 are distinct antigens recognized by antibodies in sera from immune rabbit or human cervical carcinoma patients. In addition, monoclonal antibodies to the HSV-2-induced ribonucleotide reductase were reactive with ICP10. Antibodies in sera from rabbits immunized against ICP10 and monoclonal antibodies to the HSV-2-induced ribonucleotide reductase were reactive with antigens on the plasma membrane surface of HSV-2 infected cells. Also, the reactivity of monoclonal antibodies with these antigens was blocked by the rabbit antibodies based on immunofluorescence analyses. These data provide evidence that ICP10 is antigenically distinct from ICSP11/12, and that ICP10 is present on the plasma membrane of HSV-2-infected cells. Also, our data confirm and extend the tentative identification of ICP10 with the HSV-2 induced ribonucleotide reductase recently suggested by Bacchetti et al. (J. Virol. 49, 591-593, 1984). PMID- 2988231 TI - Wilms' tumor associated with Moyamoya disease: a case report. AB - A high incidence of congenital malformations with Wilms' tumor had been recognized lately. Aniridia, hemihypertrophy, chromosome deletion, urinary tract anomalies, and Wiedemann-Beckwith syndrome have often been observed in patients with Wilms' tumor. However, Wilms' tumor associated with Moyamoya disease, which refers to a stenosis of the distal portion of the carotid artery with an abnormal network in the brain base, has never been reported. In this report, a male child aged 28 mo. with Wilms' tumor associated with Moyamoya disease is presented. PMID- 2988233 TI - [Demonstration of antibodies against tumor-associated antigens in the serum patients with bronchogenic carcinoma using anticomplement immunofluorescence on xenotransplants of human tumors]. AB - 209 sera of tumor-bearing patients and 133 control sera were tested by anti complement immunofluorescence (ACIF) on 2 xenotransplants of human bronchogenic cancer and on 2 xenotransplants of rectum carcinoma passaged in nu/nu mice. Testing of sera from patients with bronchogenic carcinoma resulted in a positive reaction with the xenotransplants of bronchogenic cancer in 48.5 to 58.5%, and sera of patients with digestive tract cancer in 6.6 to 16.1%. On xenotransplants of rectum carcinoma 73.2 to 78.1% of the sera from patients with bronchogenic carcinomas and 22.6 to 29.1% of the sera of patients with digestive tract cancer reacted positively. Control sera were positive dependent on the xenotransplants in 2.0 to 12.5%. These results indicate that there are antibodies in the sera of tumor-bearing patients against one or more non organ specific tumor-associated antigen(s) occurring in an elevated concentration in carcinomas of the digestive tract than in bronchogenic carcinomas. After absorption of the sera with homogenates of bronchogenic carcinoma and liver metastases of colon carcinoma the reaction was inhibited in the ACIF, reaching control values. PMID- 2988234 TI - [Treatment results of patients with granulosa and theca cell tumors and their mixed forms]. AB - Clinical and pathological dates of 28 patients with granulosa- and thecacelltumors and mixed granulosatheca tumors were analysed. The present therapeutical possibilities, clinical courses and the survival rate (63.6%) are described. The treatment of granulosa tumors and mixed granulosathecatumors first of all consists in hysterectomy with salpingo-oophorectomy with following irradiation. Conservative surgical therapy is only practised on young women wanting a baby later on. The connection of hormonal active ovarian tumors and the various alterations of endometrium is pointed out. An endometrial carcinoma was diagnosed only in one case. PMID- 2988235 TI - Attenuation of type A7 and A21 coxsackie viruses by transfection of cells with viral RNA. AB - Coxsackie viruses of types A7 and A21 were produced in HEp-2 cells and concentrated by precipitation with polyethylene glycol. The viral RNA was extracted by means of the hot phenol-SDS method and used for transfection of Hep 2 cells. Transfection was greatly enhanced by the addition of DEAE-dextran. 8 of 11 (72.7 per cent) coxsackie viruses obtained after 1 to 3 transfections were found to have drastically reduced pathogenicity in suckling mice in comparison to the corresponding viruses produced by infection. Attenuation was generally obtained after the first transfection and could be improved by further transfections in one experiment, only. PMID- 2988237 TI - [Pathogenesis and clinical aspects of Aujeszky's disease in cattle following an experimental infection through the respiratory, digestive and genital organs and through the skin]. PMID- 2988236 TI - [Virus infections in fishes: etiology, diagnosis and control]. PMID- 2988238 TI - Rotavirus infections in conventional pigs: kinetics excretion in faeces of rotavirus antigens, antibodies and immune complexes by pigs from birth up to three month of age. PMID- 2988239 TI - [Classification of diseases of the peripheral nervous system and formulation of the diagnosis]. PMID- 2988240 TI - [Manual therapy of patients with lumbar osteochondrosis with neurologic syndromes]. AB - Manual therapy without any additional therapeutic means was given to 325 patients with reflex syndromes of lumbar osteochondritis in an acute stage or in the stage of exacerbation of recurrent osteochondritis. A series of 454 patients in the stationary stage of exacerbation of the disease with a chronic recurrent course received a multiple modality treatment including manual therapy. Efficacy of the treatment in both groups of the patients considerably exceeded that in patients treated with the conventional (medicamentous and physiotherapeutic) methods. A considerable economic effect was achieved as a result of reducing the period of treatment and cutting down on drugs in a substantial part of the patients. PMID- 2988241 TI - [Differential roentgenodiagnosis of congenital craniocerebral hernias]. AB - Ninety-three patients with congenital craniocerebral hernia were subjected to clinical and X-ray examination. Signs characteristic of each type of congenital craniocerebral hernias were detected. Congenital craniocerebral hernia was combined with other developmental anomalies of the skull and brain in 31% of cases. Differential X-ray diagnosis is made between congenital craniocerebral hernias and traumatic birth hernias, traumatic midline hernias, cholesteatoma, dermoids and epidermoids of the orbit, polyps, papilloma, and fibrous tumors. It was established that X-ray examination plays a very important role in disclosing the bone hernial canal. The surgeon must have an exact idea of this canal before undertaking operation for plastics of the hernial defect. PMID- 2988242 TI - The interaction of metal ions with nucleic acids. Ternary complexes of copper(II) with peptides and nucleosides. AB - Difference electronic absorption and electron paramagnetic resonance spectroscopy were used to monitor the formation of the ternary complexes of Cu(II) ions with nucleosides and dipeptides containing Gly, Leu and Trp residues. Stability constants of these mixed-ligand complexes of Cu(II)-peptides with nucleosides were found to decrease in the following order: 6-ketopurines greater than 6 aminopurine greater than pyrimidines. Interpretation of the EPR data indicated that the covalent nature of the copper-ligand bond also decreases in the same order. The EPR findings suggest that nucleosides are bonded in the equatorial position of the Cu(II)-peptide complexes, however, in the case of pyrimidine nucleosides weak axial bonding also seems to occur. PMID- 2988243 TI - Polyprenol content in primary human liver carcinoma. PMID- 2988244 TI - [Tumor of the right atrium revealing a hepatocellular carcinoma]. PMID- 2988245 TI - Drugs recently released in Belgium: ceftazidime, flecainide acetate and varicella oka-strain vaccine. PMID- 2988246 TI - Application of immuno-electron microscopy to the cytologic study of benign and malignant mammary lesions. With special reference to the carcinoembryonic antigen localization patterns. AB - We investigated the application of immuno-electron microscopic (EM) observation of carcinoembryonic antigen (CEA), one of the surface markers of neoplastic epithelium, to the cytologic diagnosis of mammary cells in smears and imprints. Smears were stained by the indirect peroxidase-labeled antibody method after a modified Karnovsky fixation (4% paraformaldehyde containing 0.05% glutaraldehyde). In smears, CEA was immunocytochemically demonstrated on the entire cell membrane of malignant mammary cells whereas CEA was observed only on microvillous surfaces of benign mammary cells. Another characteristic immunocytochemical feature of malignant cells was the appearance of so-called intracellular lumina, of which the microvillous surfaces were CEA positive. These findings demonstrated that electron microscopic observation of the immunocytochemical localization of CEA is a quite useful tool for the cytologic diagnosis and characterization of neoplastic mammary lesions. PMID- 2988247 TI - Crush preparations of lesions of the central nervous system. A useful adjunct to the frozen section. AB - A series of 32 cases in which crush preparations were used in addition to frozen sections for the rapid diagnosis of lesions of the central nervous system (CNS) is presented. The cytopathologic features in crush preparations of astrocytomas, glioblastomas multiforme and a pituitary adenoma are described. Excellent preservation of cellular detail was seen in the crush preparations. Frozen sections lacked cytologic detail but provided a better view of the tissue architecture. The crush preparations yielded the correct diagnosis in 29 of the 32 cases. In the other three, a secondary component of the neoplasms (oligodendroglioma and fibrosarcoma) was identified only in the paraffin sections. Use of both frozen sections and crush preparations is recommended for all cases in which an immediate diagnosis of a CNS lesion is required. PMID- 2988249 TI - Salivary duct carcinoma of the parotid gland. Cytologic and histopathologic study. AB - Ductal carcinoma is an uncommon tumor of the salivary glands. Histopathologically, it is characterized by the presence of intraductal and infiltrative neoplastic components, morphologically resembling mammary carcinoma. A case of ductal carcinoma of the parotid gland, in which preoperative fine needle aspiration biopsy was performed, is presented. Cytologic examination of the aspirate revealed naked nuclei featuring anisokaryosis, chromatin clumps and clear vacuolar zones. PMID- 2988248 TI - Cytology of metastatic neuroendocrine (Merkel-cell) carcinoma in pleural fluid. A case report. AB - A case of Merkel-cell (neuroendocrine) carcinoma of the skin with extensive metastases, including pleural effusion, occurring over 20 years after primary resection and treatment, is reported. The histologic appearance of the primary neoplasm was identical to that seen in the biopsy specimens of the metastatic carcinoma involving the great toe and inguinal lymph nodes and to that of the residual neoplasm tissue found at necropsy. Electron microscopic examination of a lymph node metastasis demonstrated cytoplasmic microfilaments and numerous dense core, peripheral, neurosecretory granules, as previously described in Merkel-cell carcinoma. Cytologic examination of a pleural fluid specimen demonstrated numerous small malignant cells closely resembling the cells seen in the histopathologic sections from the surgical and necropsy tissues involved by metastatic carcinoma. This is the first report of the cytologic findings in a patient with Merkel-cell carcinoma metastatic to the pleural cavity. PMID- 2988251 TI - Adenylate cyclase activity in mononuclear leucocytes from patients with atopic dermatitis. PMID- 2988250 TI - Viral infections in atopic dermatitis. AB - In a study of almost 1000 patients with past or present atopic dermatitis (AD) it was found that histories of recurrent (greater than 5 episodes/year) cold sores and upper respiratory infections, as well as histories of zoster were significantly more common in AD patients than in non-atopic controls. Serological studies revealed that AD patients have clearly elevated titers of antibodies against Epstein-Barr virus. These findings suggest that the increased susceptibility to viral infections in AD is due to immune dysfunction rather than to cutaneous alterations which are associated with the disease. The mechanisms underlying the increased susceptibility to infections may be related to immunological aberrations that are secondary to a basic abnormality in the fatty acid or cyclic AMP metabolism. PMID- 2988252 TI - Lymphocyte and monocyte localization of altered adrenergic receptors, cAMP responses, and cAMP phosphodiesterase in atopic dermatitis. A possible mechanism for abnormal radiosensitive helper T cells in atopic dermatitis. PMID- 2988254 TI - Fate of receptor-bound human chorionic gonadotrophin in pseudopregnant rat ovaries perfused in vitro. AB - Pseudopregnant rats were injected with [125I] hCG, anaesthesized 1 h later and after cannulation of the aorta the ovaries were isolated and perfused with Gey & Gey buffer containing 0.2% BSA. The release of radioactivity was monitored for 2 h and analyzed by gel filtration. Five to ten per cent of the radioactivity was released within 2 h and represented small molecular weight peptides and iodotyrosine and [125I]hCG. Analysis of the ovarian radioactivity prior to and after perfusion revealed that virtually all hCG was receptor-bound. Loading the medium with unlabelled hCG displaced [125I]hCG from the receptor but did not enhance its degradation. Histological examination showed that the ovarian tissues were still intact after the 2 h perfusion. Immunohistochemical studies revealed a localization of the hCG at the cell periphery both prior to and after perfusion. These results provide evidence showing that the rate of internalization of receptor-hCG complexes in rat luteal cells is slow in vivo. PMID- 2988253 TI - Serum antibodies against Coxsackie B1-6 viruses in type 1 diabetics. AB - Twenty-two newly diagnosed insulin-dependent diabetics and 46 control subjects have been examined for Coxsackie B1-6 serum antibodies. Evidence for CoxB4 neutralizing antibodies at titers greater than or equal to 16 and two seroconversions for this single virus type were shown in the diabetic group (31.8% of patients with titers greater than or equal to 16 in comparison with 10% of controls, p = 0.08). Reduced titers against CoxB2 and CoxB5 in diabetics were also observed. In agreement with the majority of publications, these data suggest the hypothesis that CoxB4 infections are frequently associated with type 1 diabetes, and the positive relationship found only for this type of virus may be related to greater tropism for the pancreas. The specificity of this viral infection, as shown by our serologic results, could suggest a possible causative role in the development of type 1 diabetes only in a limited number of cases. The authors propose new studies with monoclonal and IgM specific antibodies, obtained from new isolates, to detect more accurately the qualiquantitative differences between type 1 diabetics and controls. PMID- 2988255 TI - Relaxin-induced changes in adenosine 3',5'-monophosphate levels in the human cervix. AB - The effects of porcine relaxin on the levels of cAMP in human cervical tissue were studied in vitro. The specimens were obtained by needle biopsy from women undergoing hysterectomy, legal abortion in the first trimester or elective Caesarean section at term, and were incubated in Krebs-Ringer buffer for 15 min in the presence of porcine relaxin (5 micrograms/ml, 3000 GPU/mg). cAMP was determined using a modified protein binding assay. The concentration of cAMP was higher in pregnant than in non-pregnant women. Relaxin stimulated the production of cAMP in the 7th-8th week of gestation and at term but did not significantly alter the cervical cAMP levels in neither non-pregnant women nor in women in the 10th-12th week of pregnancy. Previous studies have shown that porcine relaxin reduces collagen synthesis in tissue from the human cervix and lower uterine segment. The present observations indicate that these effects can be mediated by cAMP. PMID- 2988256 TI - Effect of enzyme and enzyme inhibitors on specific binding of hGH to human peripheral lymphocytes. AB - A recently described method to investigate growth hormone (GH) receptors on circulating human blood cells has been used to study the effect of a trypsin preparation and antitrypsin moieties on hormone binding to lymphocytes. Trypsinization with one defined dose of trypsin (10 ng/ml) led to a considerable decrease both of specific binding and of binding affinity (affinity constant after 60 min trypsinization 0.5 X 10(9) vs 1.5 X 10(9) M-1 in untreated control cells). Exposure of peripheral blood lymphocytes (PBL) to antitrypsin activities was followed by a steady increase of affinity and specific binding (affinity constant: with 10 KIU anti-trypsin 1.9 X 10(9) M-1, with 100 KIU 2.4 X 10(9) M-1, with 1000 KIU 3.6 X 10(9) M-1). This antitrypsin effect exceeds the binding values expected after blocking trypsin activities in the incubation medium. In a subset of experiments somatomedin-B (SM-B) was used as the antitrypsin moiety and was shown to increase specific GH binding to PBL in a similar manner as did antitrypsin (with 1000 ng SM-B affinity constant (Ka) 12.0 X 10(9) M-1, specific binding 9.7% of total radioactivity). It is concluded that enzymatic factors and their inhibitors including partially GH dependent moieties like SM-B modulate specific GH binding to human peripheral lymphocytes in vitro. PMID- 2988257 TI - The effect of nifedipine on CRF-41 and AVP-induced ACTH release in vitro. AB - The effect of nifedipine on CRF-41- and AVP-induced ACTH release was examined using monolayer cultured rat anterior pituitary cells and pituitary halves. Nifedipine inhibited ACTH release induced by synthetic rat CRF-41 in two systems. In pituitary halves, CRF-41 significantly stimulated both ACTH release and cyclic AMP accumulation. Nifedipine inhibited CRF-41-induced ACTH release and the inhibitory effect of nifedipine on CRF-41-induced ACTH release was accompanied by parallel decrease of cyclic AMP levels in pituitary halves. Nifedipine did not inhibit AVP-induced ACTH release in pituitary halves, and AVP did not significantly affect cyclic AMP accumulation in pituitary halves. These results suggest that CRF-41 stimulates ACTH release through the intracellular cyclic AMP system and calcium-calmodulin system which are accelerated by the influx of extracellular calcium ions. Moreover, it is suggested that the calcium required for AVP-induced ACTH release is derived primarily from intracellular rather than extracellular sources. PMID- 2988258 TI - Studies on the glucocorticoid-receptor blocking action of RU 38486 in cultured ACTH-secreting human pituitary tumour cells and normal rat pituitary cells. AB - The glucocorticoid-receptor blocking actions of RU 38486, a new compound with anti-progesterone activity, have been investigated in cultured human ACTH secreting pituitary tumour cells and normal rat pituitary cells. Pre-incubation of human pituitary tumour cells for 24 h with RU 38486 (1 microM) did not influence basal or CRF-stimulated ACTH release. RU 38486 (100 nM-1 microM) significantly overcame or prevented the dexamethasone (100 nM-1 microM)-induced inhibition of CRF-stimulated ACTH release by the cultured tumour cells prepared from 2 patients with Cushing's disease. The tumour cells of a third patient were insensitive to CRF. Pre-incubation for 24 h with 1 microM RU 38486 facilitated CRF-stimulated ACTH release significantly. Studies with cultured normal rat pituitary cells showed that the inhibiting effect of 24 h pre-incubation with 10 and 50 nM dexamethasone on CRF-stimulated ACTH release could be acutely (measured over 4 h) overruled in a dose-dependent way by RU 38486 (100 nM, 1 and 10 microM), while pre-incubation for 24 h of these cells with RU 38486 (100 nM and 1 microM) significantly attenuated the acute inhibiting effect of 1 microM dexamethasone on CRF-stimulated ACTH-release. The results of these in vitro experiments are discussed against the background of the possible therapeutic use RU 38486 in patients with Cushing's syndrome in order to block the deleterious effects of high circulating cortisol concentrations. PMID- 2988259 TI - Effect of ovine corticotrophin releasing factor, bromocriptine, and dopamine on release of ACTH and beta-endorphin in a patient with Cushing's disease. AB - Release of immunoreactive ACTH and beta-endorphin (beta-EP) in response to corticotrophin-releasing factor (CRF) and dopaminergic agents was studied in vivo and in vitro in a patient with Cushing's disease. Iv administration of synthetic ovine (o) CRF significantly stimulated plasma ACTH release, accompanied by increase of plasma cortisol levels. Oral administration of bromocriptine significantly suppressed plasma cortisol levels. Although reduced responses of plasma ACTH and cortisol to o-CRF was observed 1 month after removal of the pituitary adenoma, these normalized 6 months after operation. In vitro perifusion of the pituitary adenoma obtained by surgery revealed that o-CRF also stimulated ACTH and beta-EP release in a dose-responsive manner (10(-9)M 10(-5)M) and that dopamine suppressed their basal secretion. Gel exclusion chromatography of the perfusates showed that the predominant component of ACTH and beta-EP before and after o-CRF stimulation coeluted with standard ACTH and beta-EP, respectively. The present data suggest that o-CRF is a potent secretagogue for ACTH and beta-EP release from the human pituitary adenoma causing Cushing's disease and that ACTH secretion from certain adenomas, possibly originating from the intermediate lobe of the pituitary gland, is partly regulated by a dopaminergic mechanism. PMID- 2988260 TI - Serum angiotensin-I-converting enzyme in Hodgkin's disease. PMID- 2988261 TI - Autoradiographic analysis of adrenergic receptors in the mammalian brain. AB - Noradrenaline (NA) exerts its physiological and pharmacological effects in the central nervous system by interacting with specific receptor sites which are divided into four subtypes, namely alpha-1, alpha-2, beta-1 and beta-2 adrenoceptors. Alpha-1 and beta-1 receptors are thought to be neuronal and post synaptic, whereas alpha-2-R are neuronal pre- and postsynaptic and beta-2-R have a non neuronal (glial, vascular) localization. The autoradiographic localization of adrenergic receptors is requisite to a better understanding of adrenergic modulation in the nervous system. It complements analyses of adrenergic fibers and terminals and allows comparisons between afferent transmission and various receptor systems. In addition, receptor autoradiography is a preliminary step towards non invasive, in vivo receptor imaging using positron emission tomography (PET). Classical autoradiographic methods using tritium-labeled ligands are relatively tedious, as they require exposure times of several months. In order to circumvent these difficulties, an autoradiographic procedure was developed for visualization of I-125-labeled ligands. The method is validated by its application to the analysis of neuronal postsynaptic (alpha-1 and beta-1) adrenoceptors, in ferret visual cortex, in the forebrain of normal and reeler mutant mice and in the embryonic mouse brain. Distributions of alpha-1 and beta-1 adrenoceptors are studied using revelation of HEAT and ICYP binding sites, respectively. The cerebral cortex of ferret was chosen because it is widely used in vision research. The density of both alpha-1 and beta-1 adrenoceptors shows laminar heterogeneities. Beta-receptors are most heavily concentrated in cortical layers I, II and III, but very low in layer IV and moderately represented in layers V and VI. In contrast, alpha-1 receptors are more diffusely distributed, although preferentially concentrated in layer IV and, to a lesser extent, in upper cortical laminae. The two adrenoceptors are thus segregated in the radial dimension of the cortex, following distributions which are nearly complementary. These observations suggest that alpha- and beta-adrenoceptors might be associated with different stages and/or modes of information processing in the primary visual area. Adrenoceptor distribution was mapped in normal and reeler mice, in order to correlate receptor patterns with architectonic anomalies known to exist in reeler mutant mice. In normal mice, beta-1-receptors predominate in striatum, cortical layers I to III, hippocampal regio superior and some thalamic nuclei; they are moderately concentrated in cortical layers V and VI and poorly represented in lamina IV.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2988262 TI - Membrane-bound 2',3',-cyclic nucleotide 3'-phosphohydrolase activity of lymphocytes, granulocytes and erythrocytes in multiple sclerosis. AB - A sensitive fluorimetric method was used for the assay of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) activity of lymphocyte, granulocyte and erythrocyte membranes from patients with multiple sclerosis (MS). The data obtained were compared with the corresponding data from normal individuals. CNP activities of granulocyte and erythrocyte membranes of the 2 groups did not differ significantly. However, a 40% decreased membrane CNP activity of MS lymphocytes was found when the data were compared with the normal lymphocytes' activity (P less than 0.0005) by both the non-parametric median test and Student's t-test. A role of CNP in immunoregulation is suggested. PMID- 2988263 TI - Ocular abnormalities in the fetal alcohol syndrome. AB - Thirty children suffering from the fetal alcohol syndrome (FAS) were compared with 22 matched controls regarding malformations of the eyes and the visual function. All mothers to the FAS children had a documented abuse of alcohol during pregnancy. It was not possible to get reliable, detailed data on drinking habits, nor could it be ruled out that some mothers might have used psychopharmaceutic drugs during periods of the pregnancy. Defects of the outer eye region, or intraocular abnormalities, or both, were found in 27 FAS children (90%). Anomalies of the outer region like ptosis and strabismus were frequently found in the FAS children but were not common in the controls. Abnormalities in the anterior segments and media were found in six eyes. Malformations of the fundus were the most frequently occurring abnormalities in the FAS children. Forty-eight per cent of the eyes showed hypoplasia of the optic nervehead and 49% had an abnormal tortuosity of the retinal arteries. These variables were studied quantitatively by a new photogrammetric method. There were serious consequences to the vision of the FAS children. Nineteen per cent of the eyes has a visual acuity of 0.2 (20/100) or less, and 46% had 0.3-0.6 (20/60-20/30). Almost all controls had normal vision. As maternal alcohol abuse during pregnancy was the only variable which could be traced in every case of FAS, it is appears very likely that alcohol caused the abnormalities of the eyes, possibly through non specific, intra-uterine growth retardation. It that were the case, ocular abnormalities should be common in children with non-FAS growth retardation. A control study did not support this possibility. PMID- 2988264 TI - Ocular complications in insulin treated diabetes mellitus. An epidemiological study. PMID- 2988265 TI - Smooth and skeletal muscle myosins in spindle cell tumors of soft tissue. An immunohistochemical study. AB - Histological localization of myosins of the smooth and skeletal muscles was investigated in comparison with that of myoglobin by immunoperoxidase technique using the antibody against each of them in surgical specimens from spindle cell tumors and tumor-like lesions of the soft tissue. Skeletal muscle myosin was demonstrated in all of the cases of rhabdomyosarcoma, whereas myoglobin was found in 75% of the examined cases. Smooth muscle myosin was widely distributed not only in the tumor cells of smooth muscle origin such as leiomyosarcoma and angioleiomyoma, but also in the tumor cells showing myofibroblastic differentiation such as malignant fibrous histiocytoma and in the epithelial components of synovial sarcoma. The results showed that skeletal muscle myosin can be regarded as an excellent marker in the diagnosis of rhabdomyosarcoma and that smooth muscle myosin is a useful marker of leiomyosarcoma and leiomyomas, and of tumors with myofibroblastic differentiation. PMID- 2988267 TI - Xanthogranuloma of the rectum. AB - A 46-year-old man with a suspected malignant submucosal tumor received a transabdominoperineal rectal amputation. The histologic diagnosis was xanthogranuloma. The differentiation of xanthogranuloma from malignant fibrous histiocytoma and other histiocytic tumors is difficult in the preoperative stage. Malignant fibrous histiocytoma can be diagnosed histologically by the presence of pleomorphism, mitotic activity, hyperchromatism, and a storiform pattern of cell arrangement. Moreover, some xanthogranulomas are also thought to have malignant potentiality and surgical resection is regarded as the preferred treatment. As for prognosis, the patient has lived for three years after the operation but further follow-up is thought to be necessary. PMID- 2988266 TI - Multiple primary malignant fibrous histiocytoma of the stomach and small intestine. AB - A case of multicentric malignant fibrous histiocytoma of the stomach and small intestine is reported. The patient was a 60-year-old man who had total gastrectomy under an impression of a gastric carcinoma. The resected stomach revealed a large polypoid mass in the antral portion at the greater curvature. Three months later, he developed ileus and an 80 cm segment of the jejunum was removed. It contained two polypoid masses identical to that seen in the stomach. The tumors showed, in addition to the characteristic light microscopic appearances, strong positivity for alpha-1-antitrypsin by an immunoperoxidase technique, indicating the diagnosis of malignant fibrous histiocytoma (MFH). Electron microscopic findings were also consistent with MFH. We believe that this is the first well-documented case of MFH arising from the stomach and small intestine, to the best of our knowledge. PMID- 2988268 TI - Progressive multifocal leukoencephalopathy. A case report with special reference to SV40 etiology. AB - Clinical, light- and electron microscopic, and immunohistochemical findings of a 44-year-old woman with progressive multifocal leukoencephalopathy were presented. Autopsy revealed a wide distribution of the demyelinating lesion in the cerebrum, cerebellum, brain stem and spinal cord, and intranuclear inclusion bodies and papova-like virions in transmission electron microscopy in the nuclei of oligodendrocytes. SV40 antigen was immunohistochemically detected in these inclusion bodies. The widespread extension of the lesions seemed to correlate with the duration of the patient's illness. The prolongation of the clinical course in this case may be dependent upon the lack of serious underlying diseases except for a small nodule of thyroid carcinoma, SV40 infection rather than JC virus infection and/or improved care of that kind of patient. PMID- 2988269 TI - Polyclonal plasma cell proliferation with systemic capillary hemangiomatosis, endocrine disturbance, and peripheral neuropathy. AB - An autopsy case of so-called plasma cell dyscrasia (PCD) of a 37-year-old woman was reported. This case showed typical clinical symptoms of PCD, except for no abnormality in serum immunoglobulins. Besides the typical histological findings often observed with PCD, the autopsy revealed an additional finding of systemic angiomatous proliferation of small blood vessels throughout the body, especially in the skin, kidneys, brain, various lymph nodes, and soft tissue. A peculiar glomerular lesion in this case, similar to membranoproliferative glomerulonephritis, was likely to be a result of proliferation of glomerular capillaries. In addition, this case showed histological evidence of retroperitoneal fibrosis and osteosclerosis. These findings were also considered to be due to the vascular abnormality. Although the etiology of the vascular lesions is obscure, it is emphasized that the abnormality is probably an important cause of the clinical symptoms seen in PCD. PMID- 2988270 TI - Lysosomal changes in skeletal muscles during the repair of exercise injuries in muscle fibers. AB - Lysosomal changes were recorded in the skeletal muscles of mice and rats during the repair of muscle fiber injuries caused by a single bout of prolonged running. One purpose of the study was to characterize cellular and compartmental distributions of lysosomal enzymes and to investigate ultrastructural changes in the lysosomal system associated with the appearance and repair of muscle fiber injuries. Furthermore, the level of muscle fiber injuries was correlated with the lysosomal enzyme response and the indices obtained were utilized in the evaluation of the pathogenesis of exercise myopathy. The main results were: Heavy physical exertion caused scattered necrotic injuries of muscle fibers and inflammation in several skeletal muscles. The most susceptible muscles to exercise injuries were the red deep parts of quadriceps femoris muscle and the soleus and tibialis anterior muscles. The total activities of several lysosomal acid hydrolases, in particular those of beta/-glucuronidase, cathepsin C and arylsulphatase, strongly increased during the repair of exercise injuries. The highest responses occurred 3-5 days after exertion and the degree of enzyme responses correlated significantly with the level of histological injuries. The histochemical staining intensities of beta-N-acetylglucosaminidase and beta glucuronidase were highest 3 and 5 days after exertion. An increased staining intensity occurred in the inflammatory phagocytes and in the surviving muscle fibers, especially in the red muscle fibers close to necrotic foci. An autophagic response occurred in the muscle fibers close to necrotic foci 2-7 days after exertion. Autophagic vacuoles were frequently small and contained different cellular structures at various stages of degradation. The number of lysosome-like bodies, Golgi complexes and multilamellar bodies also increased. Macrophages removed the debris of necrotic muscle fibers by heterophagic uptake 2-5 days after exertion. The unsedimentable and releasable activities of acid hydrolases increased more prominently than those of the total activities in homogenates and reached their highest values 3 days after exertion. The proportional distribution of various acid hydrolases to unsedimentable, releasable and bound fractions varied strikingly but remained appreciably stable throughout the exercise myopathy. The content of lysosomal phosphomannosyl-enzyme receptors in the membrane fraction was unchanged 0-3 days after exertion but a small increase occurred later. The endogenous receptor-bound activity of beta-N acetylglucosaminidase was considerably increased 1-5 days after exertion but decreased later to the control level.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2988271 TI - Scalp hypothermia in the prevention of chemotherapy-induced alopecia. AB - Alopecia is a common side effect of cancer chemotherapy, especially in combination with regimens with doxorubicin (Adriamycin). The effect of scalp hypothermia in connection with chemotherapy was evaluated as hair protection in 61 women with disseminated breast carcinoma, where earlier treatment routines had caused wig-requiring alopecia in nearly all patients. The cooling was performed with a gel-helmet (Hypotherm Gel-Kap). Of the 61 patients, 47 (77%) had no or slight, not wig-demanding hair loss, and 14 (23%) had severe (wig-demanding) hair loss. Seven patients had liver dysfunction; in 5 of these severe hair loss was observed; 2 had slight hair loss. Eighty-three per cent of the patients with normal liver function had no hair loss. Treatment tolerance was found to be good, and side effects were minimal. The method is found to be simple, effective and inexpensive, though still not technically optimal. PMID- 2988272 TI - 3H-thymidine autoradiography and cytophotometric analysis of needle aspirates from human tumours during radiation therapy, endocrine therapy and chemotherapy. AB - Cell kinetic variables in human tumours were investigated before and during therapy. Tumour cell material was collected by sequential thin needle aspiration biopsy. Small variations in DNA replication could be detected by 3H-thymidine autoradiography. In tumours with a high rate of replication, therapy induced changes could also be detected with flow cytometry and static cytophotometry. Endocrine treatment with glucocorticoids, radiation therapy, and chemotherapy were associated with reduced rates of DNA replication in malignant lymphomas. The conformity between labelling index and S-phase estimates from DNA histograms was poorer in biopsy specimens collected during therapy than in pretreatment specimens. PMID- 2988273 TI - Blood lymphocyte population 4 to 6 years after adjuvant cyclic chemotherapy for operable breast carcinoma. AB - The influence of postoperative cyclic chemotherapy for breast carcinoma on the blood lymphocyte population was examined in 21 recurrence-free women 4 to 6 years after completion of therapy. Chemotherapy consisted of combinations of chlorambucil, methotrexate and 5-fluorouracil or cyclophosphamide, methotrexate and 5-fluorouracil. It was observed that the lymphocyte numbers and mitogenic responses to PHA and in an MLC, known to be depressed during and shortly after treatment, had recovered to the levels of healthy age-matched women. NK activity, which is increased during the period of chemotherapy, still remained elevated. PMID- 2988274 TI - Hemangioma of the vertebral column. A report on twenty-three patients with special reference to functional recovery after radiation therapy. AB - Twenty-three patients with severely symptomatic hemangiomas of the vertebral column were treated by irradiation. Pain in the back and numbness in the limbs were markedly or completely relieved in most of the patients (88% and 80%, respectively). Five out of 7 paraplegic patients recovered sufficiently to be able to walk again. For patients with a severe compression syndrome of the spinal cord, irradiation could be chosen as the primary treatment without preceding surgical decompression. The result of multiple treatment courses with lower doses was not superior to a single course, for which an optimum dose of 30 Gy to 40 Gy/4 to 6 weeks is recommended by the authors. PMID- 2988275 TI - Invasive squamous cell carcinoma of the uterine cervix. IX. Construction of a partial index in a histopathologic malignancy grading system. AB - A malignancy grading system (MGS) for invasive squamous cell carcinoma of the uterine cervix, consisting of four items for the tumour-cell population and four items for the tumour-host relationship, proved to have a good prognostic capacity. A correlation analysis based on 161 patients with invasive squamous cell carcinoma of the uterine cervix in stages I and II was evaluated. The relationships between survival/lethality ratio (S/L) and all single items, MGS, and a reduced partial index were compared. The study showed good agreement with previous studies on another partial index. The MGS had the best predictive value. The items of the tumour-cell population had no significant predictive value. According to a regression analysis, items for the tumour-host relationship or the items vascular invasion and host-cellular response alone could be used without any substantial loss of predictive capacity compared with the MGS. However, the MGS remains the most complete and adequate classification for invasive squamous cell carcinoma of the uterine cervix. PMID- 2988276 TI - Intracavitary irradiation of carcinoma of the cervix stage IB and IIA. A clinical comparison between a remote high dose-rate afterloading system and a low dose rate manual system. AB - The clinical results of a remote high dose-rate afterloading technique with individual three-dimensional treatment planning were compared with those of a manual low dose-rate technique for intracavitary irradiation of cervical carcinoma stage IB and IIA. The rates of residual tumour at operation, local recurrence and survival were comparable with the two techniques. The rate of treatment complications was lower with the high dose-rate technique even when external irradiation was added. Better central shielding for external pelvic irradiation is possible when intracavitary irradiation is performed with higher accuracy. PMID- 2988277 TI - Cohort study of the long-term effect of irradiation for carcinoma of the uterine cervix. Second primary malignancies in the pelvic organs in women irradiated for cervical carcinoma at Radiumhemmet 1914-1965. AB - The risk of second primary malignancy arising after therapeutic irradiation was evaluated in the Radiumhemmet series of carcinoma of the uterine cervix treated in 1914-1965. Only tumours appearing more than 10 years after irradiation were taken into account. Comparisons of observed with expected incidence showed excess of malignancies in urinary bladder, endometrium, ovaries and rectum, but not of colon carcinoma. Comparisons were made with cohorts from the Swedish Cancer Registry. PMID- 2988278 TI - Prognostic factors in stage I and II non-Hodgkin's lymphoma of Waldeyer's ring. AB - Sixty-nine stage I and II patients treated for non-Hodgkin's lymphoma of Waldeyer's ring were retrospectively analysed. Diffuse histiocytic lymphoma (Rappaport's classification) was the most common histology (67%). Staging without laparotomy revealed 31 patients in stage I and 38 in stage II. Sixty-five patients received radiation therapy against involved or extended fields. In 43 patients adjuvant chemotherapy with single or multiple agents was given and 4 patients received only chemotherapy. A relapse occurred in 34 (49%) patients. The 5-year relapse-free survival was 67 per cent and 36 per cent in stage I and stage II, respectively. Radiation doses of 50 Gy gave no recurrence within treated volumes. However, since relapse below the clavicles and especially below the diaphragm was common, the radiation doses did not seem important for reduction of the overall failure rate. In stage II, patients with a single neck node less than 5 cm in diameter seemed to have a better prognosis than patients with large or multiple neck nodes. In this non-randomized study adjuvant chemotherapy seemed to reduce the risk of relapse, especially in stage II patients. PMID- 2988279 TI - Immunodeficiency and prognosis in patients with non-Hodgkin's lymphomas. AB - Monocyte depleted blood lymphocyte subpopulations, their functions and relation to prognosis were studied in 68 untreated adult patients with non-Hodgkin's lymphomas classified according to the Kiel nomenclature. The median observation time was 48 months (range 41-60). The mean total blood lymphocyte and T (SRBC rosetting) cell counts were significantly decreased as compared with age-matched controls (n = 57). Twenty-five per cent of the patients had a monoclonal blood B lymphocyte population. The spontaneous lymphocyte DNA synthesis, measured as incorporation of 14C-thymidine, was increased and the response to mitogen and antigen stimulation was decreased. Blood lymphocyte counts and functions before treatment were not related to the rates of remission or survival. PMID- 2988280 TI - Serum thymidine kinase as a prognostic marker in Hodgkin's disease. AB - In 72 of 81 consecutively diagnosed patients with Hodgkin's disease pretreatment sera were available for analysis of serum thymidine kinase (S-TK) levels. In relation to clinical parameters, such as stage, histopathology and general symptoms, significant correlations were found with higher (S-TK) levels in advanced disease as well as in patients with B-symptoms. When the prognostic ability was examined, patients in stages IA and IIA could be divided according to S-TK levels into two different groups in relation to disease-free survival. This latter finding makes this serum test interesting as an additional tool in the clinical evaluation and in the therapeutic decision concerning patients with Hodgkin's disease. PMID- 2988281 TI - Totally implanted device for venous access. Experience in tumour patients. AB - A device for central venous access consisting of a subcutaneous injection portal (Port-A-Cath) connected to a silicone rubber catheter was implanted in 30 tumour patients. The system was used mainly for the administration of antineoplastic drugs. During the investigation period (18 months) the systems remained in function for a median time of 155 days (range 7-372 days). Drug injections were given to 22/30 patients on a median of 11 occasions (range 2-65). Continuous infusions were administered to 17/30 patients for a median time of 15 days (range 1-43). Routine flushing with heparinized saline was performed monthly. Ten patients died with the system still functioning. In another 18 cases the devices were working at the completion of the study. One catheter extravasated. Skin erosion at the site of the portal occurred in 2 cases, in one of which the system had to be extracted. Partial caval thrombosis developed around a catheter tip in one case. No system became occluded or infected. Overall patient acceptance was excellent. PMID- 2988282 TI - Accumulation of 99Tcm-gluconate in daunorubicin-treated neonatal heart cells in culture. AB - The cardiotoxic effect of daunorubicin was investigated using cultured heart cells from neonatal rat hearts. Exposure of the cells to low but toxic concentrations of daunorubicin led to drastically altered permeability properties of the plasma membrane as measured by release of lactate dehydrogenase. Simultaneously, 99Tcm-gluconate, an isotope complex used in scintigraphic imaging of myocardial infarcts, was accumulated in the cells. The results suggest that daunorubicin-induced cardiac damage may be estimated by isotope uptake. PMID- 2988283 TI - Interactions of radiation and cancer chemotherapeutic drugs in a C3H mouse mammary carcinoma. AB - The interactions of radiation and adriamycin (ADM), bleomycin (BLM), cyclophosphamide (CTX), 5-fluorouracil (5-FU), methotrexate (MTX), mitomycin C (MM-C), or cis-diamminedichloroplatinum II (cis-DDP) were studied in a spontaneously arisen C3H mouse mammary carcinoma. The tumour response to drugs alone was evaluated by measuring the tumour to reach a volume 5 times that of the treatment day. CTX resulted in a marked tumour growth delay whereas the other drugs had a modest or uncertain effect. In the combined treatment experiments, drugs were administered as single doses either 15 min before or 4 hours after graded single doses of irradiation. The end point for each treatment was the radiation dose which on an average was required to achieve local tumour control in 50 per cent of the mice (TCD50). The dose effect factor (DEF) was 1.16 for ADM and 1.17 for CTX, the enhanced radiation response being independent of administration before or after irradiation. MM-C also decreased the TCD50 for radiation alone, but its effect was more marked 15 min before (DEF 1.32) than 4 hours after irradiation (DEF 1.18). BLM, 5-FU, MTX, and cis-DDP had no effect on the radiation response neither when administered 15 min before nor 4 hours after irradiation. PMID- 2988284 TI - Influence of catalase on the radiation sensitizing effect of misonidazole. AB - The radiation modifying action of misonidazole and catalase was investigated in Bacillus megaterium spores at various oxygen concentrations. Catalase (120 micrograms/ml) decreased the radiation sensitizing action of misonidazole. Misonidazole as an electron affinic radiation sensitizer enhanced the build up of H2O2, thus promoting the reaction with catalase. Protection by catalase was not enough to eliminate the total radiation sensitizing effect of misonidazole. PMID- 2988285 TI - Stimulatory effect of intermittent feeding on hemopoietic recovery in sublethally gamma-irradiated mice. AB - The effect of three-week adaptation to intermittent feeding on the recovery of the hemopoietic functions of mice after sublethal gamma irradiation was investigated. Measurement of oxygen consumption, carbon dioxide output and the respiratory quotient demonstrated an increased metabolic rate in the intermittently fed animals and an accentuation of lipogenic processes. This metabolic state persisted even after irradiation. An improvement in the recovery of hemopoietic functions after irradiation was demonstrated in adapted animals, which was reflected by the increased proliferative activity of the hemopoietic cell populations (more intensive incorporation of 125I-UdR into the DNA of cells of the spleen, thymus and femoral bone marrow), by more rapid renewal of spleen weight, more rapid recovery of the femoral bone marrow cellularity and increased levels of granulocytes in peripheral blood. PMID- 2988286 TI - Characteristics of a selectively shielded p-Si detector in 60Co and 8 and 16 MV roentgen radiation. AB - A p-silicon semiconductor detector with a filter of wolfram powder mixed with epoxy, that entirely covered the back of the detector, was investigated and compared with ionization chambers and an unshielded semiconductor detector. Relative depth and profile distributions obtained as signals from the semiconductors were compared with corresponding dose distributions measured with the ionization chambers in 60Co, 8 and 16 MV roentgen radiation of different field sizes. It was found that relative signal distributions from the shielded semiconductor detector agreed, within 1 per cent of the maximum signal, to the depth dose curves and that the relative signal in profile distributions also agreed, within 1 mm or 1 per cent of the signal at the central axis, as compared with dose measurements with a cylindrical, thimble ionization chamber. The relative signal in the building-up region was compared with a plane parallel ionization chamber with a deviation corresponding to a position of less than 1 mm. PMID- 2988287 TI - Transformation of human fetal thymus and spleen lymphocytes by human T-cell leukemia virus type I. AB - Co-cultivation of human thymus and spleen lymphocytes, which were obtained from 26-week and 27-week fetuses, with a lethally-irradiated human cord T-cell line harboring human T-cell leukemia virus type I (HTLV-I) resulted in the establishment of T-cell lines positive for adult T-cell leukemia-associated antigens and producing HTLV-I. These cell lines had the phenotype of a helper/inducer subset of peripheral T-cells as evidenced by the reactivity with monoclonal antibodies to human T-cells. PMID- 2988288 TI - [Drug activation of GABAergic transmission in the central nervous system: benzodiazepines and GABAergic agonists]. PMID- 2988289 TI - Effects of hypoxia and ischaemia on coronary vascular resistance, A-V node conduction and S-A node excitation. AB - Hypoxia and ischaemia produce relaxation of the vascular smooth muscle of the resistance vessels in the myocardium. A low pH may contribute to the arteriolar dilatation but cannot account for the major response. In all likelihood the reduced pO2 acts indirectly by release of a vasodilator mediator rather than by a direct effect on the vascular muscle. Potassium release may account for a transient component of the vasodilation, but it appears that a major factor is the release of adenosine from the ischaemic tissue. Adenosine also appears to be responsible for slowing of the sinus rate and impaired A-V conduction in myocardial ischaemia. These observations are of clinical significance because hypoxia-induced A-V block, such as may occur in inferior myocardial infarction, can be abolished by an adenosine antagonist such as aminophylline. In contrast, adenosine is also useful in the treatment of some cardiac arrhythmias. Supraventricular tachycardia, in which the A-V node is involved in the re-entrant pathway, is readily abolished in humans by the intravenous administration of small doses of adenosine. PMID- 2988290 TI - Tumour ploidy in DNA histograms of pituitary adenomas. AB - DNA analysis was performed by flow cytometry (FCM) in 59 cases of different types of pituitary adenomas who underwent microsurgery and were actively followed over a period of 3-6 years. Aneuploid DNA patterns were detected in 12 (20%) cases. Frequency of aneuploidy was highest in prolactin-secreting (36%) as compared to 17% in GH- and 7% in non-secreting adenomas. Some adenoma parameters valid for tumour growth, such as rate of recurrences and mitotic index, were correlated with aneuploidy of the tumour. FCM studies are, therefore, recommended to gain further parameters for the estimation of proliferative activity of pituitary adenomas. PMID- 2988291 TI - Trophic and myogenic effects with special reference to transferrin. PMID- 2988292 TI - Genetic analysis of Duchenne dystrophy. PMID- 2988293 TI - Biochemical characterization of the adrenergic receptors: affinity labeling, purification, and reconstitution studies. PMID- 2988294 TI - Homology between light-activated photoreceptor phosphodiesterase and hormone activated adenylate cyclase systems. PMID- 2988295 TI - Dual regulation of adenylate cyclase activity and hormone release in the intermediate lobe of the rat pituitary gland: evidence for the involvement of membrane components of the stimulatory beta 2-adrenergic system and of the inhibitory D-2 dopaminergic system. PMID- 2988296 TI - ADP-ribosylation of the inhibitory guanine nucleotide regulatory protein (Ni) as a possible mechanism underlying development of beta-adrenergic responses during primary culture of rat hepatocytes. PMID- 2988298 TI - Lipolysis and cyclic-AMP-dependent protein kinase in adipocytes: adenylate cyclase regulators versus insulin. PMID- 2988297 TI - Regulation of cyclic AMP metabolism by muscarinic cholinergic receptors. AB - The occurrence of muscarinic cholinergic receptor-mediated activation of phosphodiesterase in 1321N1 cells does not represent an isolated phenomenon, since a similar response to cholinergic stimuli is observed in thyroid slices (45) and WI-38 fibroblasts (1,42). Both muscarinic-receptor-mediated inhibition of adenylate cyclase and activation of phosphodiesterase occur in WI-38 fibroblasts (42). Work currently under way in our laboratory is directed toward determining if a guanine nucleotide regulatory protein is involved in the activation of phosphodiesterase in these cells and whether common or separate populations of muscarinic receptors are coupled to these two mechanisms of cyclic AMP metabolism. The analysis of acute hormonal regulation of phosphodiesterase in intact cells is sufficiently complicated to have previously discouraged investigators from pursuing this question in mammalian tissues. The 1321N1 cell line provides a simple model system in which at least one mechanism of hormonal regulation of phosphodiesterase can be examined. In light of the widespread occurrence of muscarinic-receptor-mediated effects on Ca2+ mobilization, it would not be surprising to find that this mechanism represents an important part of cholinergic action in both the peripheral and central nervous systems. Indeed, this system could provide an important regulatory link between Ca2+ -mediated and cyclic-AMP-mediated events in target cells. The potential importance of such a mechanism also need not be restricted to the muscarinic receptor system, since any neurotransmitter or hormone receptor system coupled to events involved in Ca2+ mobilization might produce phenomena similar to that observed for muscarinic receptors in 1321N1 cells. Our studies emphasize that two mechanisms for regulation of cyclic AMP accumulation by muscarinic cholinergic receptors exist. The data to date suggest that separate receptor subtypes are involved in these mechanisms of cholinergic regulation and provide another biochemical basis whereby the well-known interaction of Ca2+ with the cyclic AMP system can be effected. Thus, identification of the molecular events involved in the regulation of PI turnover and its consequences may be crucial in defining the basis of this aspect of cholinergic action. In addition, more extensive analyses of the phosphodiesterase system using cell-free preparations have the potential of providing clues to the molecular basis of this mechanism.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2988299 TI - Human platelet alpha 2-adrenergic receptors: effect of Na+ on interaction with the adenylate cyclase system and on epinephrine-stimulated platelet secretion. PMID- 2988300 TI - Adenosine modulation of synaptic responses in rat hippocampus: possible role of inhibition or activation of adenylate cyclase. PMID- 2988302 TI - Adenosine receptors. PMID- 2988301 TI - Altered subcellular distribution of protein kinase C (a phorbol ester receptor). Possible role in tumor promotion and the regulation of cell growth: relationship to changes in adenylate cyclase activity. PMID- 2988303 TI - Receptors coupled to calcium mobilization. PMID- 2988304 TI - beta-Adrenergic receptors and regulatory GTP-binding proteins: reconstitution of coupling in phospholipid vesicles. PMID- 2988305 TI - Properties required of a functional Ni, the GTP regulatory complex that mediates the inhibitory actions of neurotransmitters on adenylate cyclase. PMID- 2988307 TI - Epstein-Barr virus infection and immunoregulation in man. PMID- 2988306 TI - The complexity of virus--cell interactions in Abelson virus infection of lymphoid and other hematopoietic cells. PMID- 2988308 TI - Drugs, prostaglandins, and renal function. PMID- 2988309 TI - Urinary 6-keto-PGF1 alpha after captopril and indomethacin: possible contribution of PGI2 to the antihypertensive mechanism of ACE inhibitors. PMID- 2988310 TI - Thromboxane and leukotrienes in clinical and experimental transplant rejection. PMID- 2988311 TI - Eicosanoids as regulators of pancreatic islet hormone secretion. PMID- 2988312 TI - Thromboxane- and ADP-independent aggregation and exposure of fibrinogen receptors on platelets from patients with familial hypercholesterolemia. PMID- 2988313 TI - Role of prostaglandins in the activity of some cardiovascular drugs. PMID- 2988314 TI - Is prostacyclin effective on transmembrane calcium movements? PMID- 2988316 TI - [Clinico-epidemiological study of keratoconjunctivitis due to adenovirus type 37 (Ad 37) in Sapporo, Japan]. PMID- 2988315 TI - Superoxide dismutase inhibits LTB4-induced leukotaxis. PMID- 2988317 TI - Reproducibility of T1 and T2 relaxation times calculated from routine MR imaging sequences: phantom study. AB - Measurement of T1 and T2 relaxation times has been sought as one fundamental way to characterize tissue. Relaxation times can be calculated from routine spin-echo (SE) imaging sequences using two distinct repetition times (TRs), each with two SE samplings of signal intensity. Previous reports have quantified relaxation times without discussing the variation in their measurements. By imaging a phantom containing different samples with known T1 and T2 relaxation times on three separate occasions, the variation in relaxation time measurements inherent in different routine imaging sequences was studied. For the present study a more complete and accurate equation was used to calculate T1 values. The variation in T1 and T2 relaxation times for samples with relaxation times similar to solid tissue was 2%-4%. The amount of variability in calculated relaxation times was found to be dependent on the magnitude of the relaxation times themselves. However, the mean values were independent of the imaging sequences used to calculate the relaxation times. No significant differences were seen between left to-right or section-to-section position within the same study or between studies performed on different occasions. The variability in the calculated T1 was dependent on the pair of TR sequences used to calculate T1. Samples with long T1 and T2 relaxation times, similar to many body fluids, had much larger variability. A computer simulation of measurement error was created to explain these results. This study indicates that properly performed routine imaging studies do yield reproducible T1 and T2 measurements. PMID- 2988318 TI - Reproducibility of relaxation times and spin density calculated from routine MR imaging sequences: clinical study of the CNS. AB - This study was undertaken to determine if routine clinical magnetic resonance imaging sequences using only two different repetition times (TRs) and with only two sequential spin echoes (SEs) can be used to calculate reproducible relaxation time and spin density values for normal central nervous system tissue using a 0.35 T production-model instrument. In 43 patients 650 regions of interest of 11 different anatomic sites were measured. T1 and T2 relaxation times and spin density were measured. For each anatomic location, the mean and standard deviation of these values were determined. In most solid regions of brain, the standard deviation of both T1 and T2 was 4%-8%. Relaxation times of cortical gray matter varied more, with a standard deviation of 10%, probably because of volume averaging with adjacent cerebrospinal fluid (CSF). CSF and ocular vitreous humor were neither reproducibly nor accurately measured because of the short TR and TE settings of the imaging sequences relative to the long T1 and T2 relaxation times of these substances. Significant and reproducible differences were found between the spin densities of gray matter and white matter, as well as between different regions of white matter. These differences are of major importance in contrast discrimination of gray and white matter on the long TR images. Knowing that relaxation values and spin densities calculated from routine imaging sequences are in fact reproducible, these normal ranges can now be used to investigate changes occurring in disease states. PMID- 2988319 TI - Cytomegalovirus esophagitis and gastritis in AIDS. AB - The radiographic features of cytomegalovirus involvement of the upper gastrointestinal tract in four homosexual men with acquired immunodeficiency syndrome (AIDS) are described. Esophageal involvement (three cases) was demonstrated as a localized ulceration or as a more diffuse esophagitis affecting predominantly the distal esophagus. Gastric involvement (two cases) presented as large nodular rugal folds in the fundus or as a circumferentially narrowed antrum deformed by numerous large nodular contour defects. Endoscopy revealed mucosal erosions, serpiginous ulcers, and inflammatory exudate; biopsies documented the presence of typical inclusion bodies in the epithelial and endothelial cells. Although the radiographic findings are nonspecific, the diagnosis of cytomegalovirus esophagitis and gastritis should be suspected when similar abnormalities are present in immunocompromised patients, particularly homosexual men with AIDS. PMID- 2988320 TI - Benign and malignant breast disease: magnetic resonance and radiofrequency pulse sequences. AB - Thirty patients with suspected abnormality of the breasts on mammography were evaluated with magnetic resonance imaging (MRI) in a blind fashion. Spin-echo (SE) 250/30 msec scans were used to screen the examined breast. At the location of the suspected abnormality, inversion recovery (IR) 1000/30/300, SE 1000/30, and SE 1000/120 scans were performed. On the basis of these magnetic resonance images and experience with 70 previously studied patients, abnormalities of the breasts were grouped into five patterns. Ten malignant lesions exceeded 1 cm in diameter and were all correctly diagnosed by mammography and MRI. Of the remaining 20 benign conditions, four were suspicious for malignancy on MRI compared to eight with mammography. The shape of the lesion and the change in its signal intensity with different MR radiofrequency pulse sequences allows differentiation between a benign and a malignant process. On the basis of this preliminary experience, it seems MRI may have an adjunctive role to screening mammography. SE 1000/120 scans show higher signal intensity from carcinoma than from normal duct tissue, fibrocystic disease, and fibroadenoma. In this respect, it may allow some tissue specification in the breast. PMID- 2988321 TI - Computed tomography of hilar cholangiocarcinoma: a new sign. AB - Thirty-seven patients with histologic proof of cholangiocarcinoma at the confluence were examined by computed tomography (CT) to determine whether this examination is of value in the assessment of these patients for surgery and whether there are any features specific to this type of tumor. Thirty-two patients showed intrahepatic duct dilatation; six of these showed dilatation of ducts in one lobe only. Eighteen patients had intrahepatic low-attenuation areas, while eight had a mass lesion in the porta hepatis. Two of the lesions in the porta hepatis and four of the low-attenuation lesions enhanced. Atrophy of a lobe was noted in seven patients. This feature is not commonly recognized and is suggestive of cholangiocarcinoma rather than other hepatic tumors. The results of this study show that CT provides useful anatomic information preoperatively but that the appearances are nonspecific. Lobar atrophy is highly suggestive of hilar cholangiocarcinoma, either of long-standing or with unilateral portal venous involvement. PMID- 2988322 TI - "In vitro" basophil degranulation by polymyxin B in asthmatic patients. AB - Polymyxin B is a potent mast cell degranulator. Inhalation of polymyxin B produces bronchoconstriction in atopic asthmatic patients. The study was undertaken to find the influence of polymyxin B on human basophils. Thirty atopic asthmatic patients and 10 healthy controls participated in the study. Preparation and counting of basophils taken from the whole venous blood was performed with the human basophils degranulation test (method by Benveniste). No significant degranulation was observed in the group of healthy subjects. Degranulation of basophils by polymyxin B was stated in 24 from 30 atopic asthmatic patients with dose-reaction dependency. Our results point out, that there exists "basophil releasability" to polymyxin B in atopic asthmatic patients. PMID- 2988323 TI - Sub-aggregatory doses of catecholamines prevent prostacyclin-induced inhibition of platelet aggregation. AB - To assess the effect of subaggregatory concentrations of catecholamines on the antiaggregatory effect of prostacyclin (PGI2), platelets from normal human volunteers were exposed sequentially in vitro to epinephrine (less than or equal to 50 nM)- or norepinephrine (less than or equal to 1 microM) followed by PGI2 and adenosine diphosphate (ADP). Platelets thus pretreated did not manifest the normal inhibitory response to PGI2, aggregating to a similar extent as platelets exposed to ADP alone. This effect was unaffected by aspirin but was abolished by exposure to phentolamine. Catecholamine pretreatment similarly blocked the PGI2 induced increase in intracellular cyclic AMP, an effect which was also reversed by phentolamine. These data suggest that platelets exposed in vivo to elevated catecholamine concentrations, such as are seen clinically during myocardial infarction, might be similarly unresponsive to endogenous PGI2. PMID- 2988324 TI - Modified Marcali method for the determination of total toluenediisocyanate in air. AB - The Marcali method for the analysis of airborne toluenediisocyanate was evaluated for its response to the two isomers of the compound. The absorbance of the chromophore obtained from 2,6 TDI is approximately 47% less than that from the 2,4 isomer of TDI when measured at 550 nm. In addition, the absorbance spectra of the 2,6 isomer exhibits a shifting of the wavelength of maximum absorbance from 550 nm to 565 nm. A simple modification of the method involving changes in diazotization time and temperature is proposed to eliminate this isomer effect. PMID- 2988325 TI - Electrocardiographic, enzymatic and scintigraphic criteria of acute myocardial infarction as determined from study of 726 patients (A MILIS Study). AB - Methods for detecting acute myocardial infarction (AMI) were compared in a prospective study of 726 patients with pain presumed to be caused by ischemia that lasted 30 minutes or longer and was associated with electrocardiographic changes (ST-segment deviation greater than or equal to 0.1 mV and/or new Q waves or left bundle branch block). Using MB-CK values of more than 12 IU/liter as the standard criterion for detection of AMI, 639 patients (88%) were judged to have AMI. Total plasma CK values, technetium-99m stannous pyrophosphate images 48 to 72 hours after admission, and serial 12-lead electrocardiograms over 10 days were analyzed by investigators blinded to other clinical and laboratory data. For detection of AMI, total CK, electrocardiograms (ECGs) and pyrophosphate imaging were all highly accurate and sensitive (total CK accuracy 97%, ECG 92%, pyrophosphate 88%; total CK sensitivity 98%, ECG 96% and pyrophosphate 91%). However, both pyrophosphate and ECG were less specific than total CK (p less than 0.01) (total CK specificity 89%, pyrophosphate 64% and ECG 59%). The sensitivity (p less than 0.05) and accuracy (p less than 0.01) of total CK and pyrophosphate for those patients with Q-wave development were slightly greater than for those in whom Q waves did not evolve. The ECG was less accurate (p less than 0.02) and pyrophosphate was less specific (p less than 0.04) in patients with prior MI compared with those with initial infarction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988326 TI - Effect of enalapril at rest, during tilt, static and dynamic exercise in systemic hypertension. AB - Using the "Oxford system" for ambulatory monitoring of direct arterial blood pressure (BP), the hypotensive effect of enalapril (20 to 40 mg/day), was assessed in 15 patients with essential hypertension. BP was reduced by enalapril throughout the 24 hours of study. A within-patient comparison of corresponding mean hourly systolic and diastolic BP values showed a reduction from (p less than 0.05 to p less than 0.001) for 18 of the 24 hours. Four patients had an increase in BP during treatment with enalapril. There was a significant decrease in systolic BP in response to 60 degrees head-up tilt. During isometric and dynamic exercise there was a smaller increase in systolic BP but a less marked effect on diastolic BP. Heart rate was unaffected either during free ambulation or during physiologic testing. The incidence of side effects was low. These results indicate that enalapril administered once daily may be an effective form of antihypertensive therapy. However, there is a group of patients who are unresponsive to treatment with converting enzyme inhibitors. PMID- 2988327 TI - Cyclic AMP-mediated jejunal secretion in lactose-fed malnourished rats. AB - We evaluated the effects of a hyperosmolar lactose load on the functional properties of the jejunal mucosa of protein-energy malnourished rats. Malnourished animals exposed to an oral lactose load showed an enhanced loss of DNA, protein, and sodium into the intestinal lumen as compared to well nourished controls exposed to the same lactose load. Only the jejunum of lactose-fed malnourished rats showed increased levels of 3'-5' cyclic adenosine monophosphate (cAMP), that appeared principally due to an enhancement of adenyl cyclase. Overall, our data strongly suggest that the intestinal mucosa in malnutrition is more sensitive and responds to an osmotic stress by enhanced secretion. PMID- 2988328 TI - A practical cytopathic effect/dye-uptake interferon assay for routine use in the clinical laboratory. AB - The clinical value of interferon (IFN) level determinations has been demonstrated, but a practical assay procedure for routine use in the diagnostic laboratory has not been available. The authors examined the susceptibility of five cell lines (WISH, HEp-2, Vero, A549, and WI-26VA4) to vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV), and compared the response of these cells to the inhibitory activity of IFN-alpha and IFN-gamma for the respective viral cytopathic effect (CPE). The WISH-EMCV system was the most sensitive for IFN-alpha, and was approximately as sensitive as the HEp-2-VSV system for IFN-gamma. WISH cells were found to be significantly more sensitive for both IFN-alpha and IFN-gamma when EMCV, instead of VSV, was used (P less than 0.001). Therefore, the latter system served as the basis for developing a CPE dye uptake procedure that was found to be considerably more rapid but slightly less sensitive than the conventional technic. However, both procedures were equally reproducible and should be suitable for automation. PMID- 2988329 TI - Cytomegalovirus esophagogastritis in a patient with acquired immunodeficiency syndrome. AB - Symptomatic cytomegalovirus involvement of the gastrointestinal tract has become increasingly recognized in patients with acquired immunodeficiency syndrome. This may take varying forms, often mimicking more well known entities. This is a report of a male intravenous drug abuser with documented acquired immunodeficiency syndrome, who developed dysphagia and evidence of distal esophagitis. Although this was initially thought to be related to reflux peptic disease, he subsequently was documented to have cytomegalovirus esophagitis and gastritis. His course was further complicated by small bowel perforation with cytomegalovirus found here as well. Cytomegalovirus infection, in general, and its increasingly recognized role in the morbidity of patients with acquired immunodeficiency syndrome, is discussed. PMID- 2988330 TI - Cytomegalovirus colitis in acquired immunodeficiency syndrome--a chronic disease with varying manifestations. AB - A patient with acquired immunodeficiency syndrome and cytomegalovirus colitis is described. The colitis is more chronic and less severe than earlier descriptions. This case presents different and more varied endoscopic findings than previously reported. PMID- 2988331 TI - Mapping DNA sequences in a human X-chromosome deletion which extends across the region of the Duchenne muscular dystrophy mutation. AB - A somatic cell hybrid has been constructed and characterized using fibroblasts from a phenotypically normal woman who possesses an X chromosome with an interstitial deletion of the short arm. High-resolution banding indicates that the deleted segment is either Xp22.13-p11.4 or Xp22.11-p11.23. Southern blot hybridization to previously mapped DNA sequences confirms that the missing segment of the X chromosome is a deletion and not an interstitial translocation and supports the cytogenetic interpretation that the deletion extends proximal of Xp11.3 and therefore probably comprises Xp22.11-p11.23. Three further DNA sequences have been localized to the region of the deleted segment. The following order has been assigned to the seven probes used: Xpter-RC8-pXUT22-(OA1,C7,M2C) L1.28-RD6 -Xcen. PMID- 2988332 TI - Genetic mapping of DNA segments relative to the locus for the fragile-X syndrome at Xq27.3. AB - We have tested linkage between the locus for the fragile-X [fra(X)] syndrome at Xq27.3 and five polymorphic restriction sites identified by four DNA probes mapping distal to Xq26.1. A maximum distance of approximately 15 centimorgans (cM) between Xq27.3 and the marker loci mapping to this region was predicted based on the physical chromosome length. Close linkage between the disease and marker loci was excluded for probes DXS19 and DXS37 (theta = .05, Z = -2.94 and Z = -4.17, respectively). These marker loci were estimated to be less than five cM apart but approximately 40 cM proximal to the fragile site, indicating that there is a significantly greater frequency of recombination in this region of the X chromosome than expected from the physical length. Linkage results for the other marker loci and the fra(X) syndrome were inconclusive. However, the pX45d probe locus appears very closely linked to the factor IX locus (Z = 1.94 at theta = 0) and is approximately 20 cM proximal to Xq27.3. A relative map of the polymorphic restriction sites, fra(X) syndrome locus, and factor IX locus was constructed by maximizing lod scores over the Xq26.1----q27.3 region. PMID- 2988333 TI - Choroideremia is linked to the restriction fragment length polymorphism DXYS1 at XQ13-21. AB - Choroideremia (McK30310), an X-linked hereditary retinal dystrophy, causes night blindness, progressive peripheral visual field loss, and, ultimately, central blindness in affected males. The location of choroideremia on the X chromosome is unknown. We have used restriction fragment length polymorphisms from the X chromosome to determine the regional localization of choroideremia by linkage analysis in families with this disease. One such polymorphic locus, DXYS1, located on the long arm (Xq) within bands q13-q21, shows no recombination with choroideremia at lod = 5.78. Therefore, with 90% probability, choroideremia maps within 9 centiMorgans (cM) of DXYS1. Another polymorphic locus, DXS11, located within Xq24-q26, also shows no recombination with choroideremia, although at a smaller lod score of 1.54 (90% probability limit theta less than 30 cM). This linkage with DXS11, a marker that is distal to DXYS1, suggests that the locus for choroideremia is also distal to DXYS1 and lies between these two markers in the region Xq13-q24. These results provide regional mapping for the disease that may be useful for prenatal diagnosis and, perhaps ultimately, for isolating the gene locus for choroideremia. PMID- 2988334 TI - Chromosome-specific organization of human alpha satellite DNA. AB - Restriction endonuclease analysis of human genomic DNA has previously revealed several prominent repeated DNA families defined by regularly spaced enzyme recognition sites. One of these families, termed alpha satellite DNA, was originally identified as tandemly repeated 340- or 680-base pair (bp) EcoRI fragments that hybridize to the centromeric regions of human chromosomes. We have investigated the molecular organization of alpha satellite DNA on individual human chromosomes by filter hybridization and in situ hybridization analysis of human DNA and DNA from rodent/human somatic cell hybrids, each containing only a single human chromosome. We used as probes a cloned 340-bp EcoRI alpha satellite fragment and a cloned alpha satellite-containing 2.0-kilobase pair (kbp) BamHI fragment from the pericentromeric region of the human X chromosome. In each somatic cell hybrid DNA, the two probes hybridized to a distinct subset of DNA fragments detected in total human genomic DNA. Thus, alpha satellite DNA on each of the human chromosomes examined--the X and Y chromosomes and autosomes 3, 4, and 21--is organized in a specific and limited number of molecular domains. The data indicate that subsets of alpha satellite DNA on individual chromosomes differ from one another, both with respect to restriction enzyme periodicities and with respect to their degree of sequence relatedness. The results suggest that some, and perhaps many, human chromosomes are characterized by a specific organization of alpha satellite DNA at their centromeres and that, under appropriate experimental conditions, cloned representatives of alpha satellite subfamilies may serve as a new class of chromosome-specific DNA markers. PMID- 2988335 TI - Melanesians and Polynesians share a unique alpha-thalassemia mutation. AB - Several genetic markers that provide information on population migrations and affinities have been detected by studies of proteins and cellular antigens in blood. Analysis of DNA polymorphisms promises to yield many further population markers, and we report here the distribution of a new alpha-globin gene deletion (-alpha 3.7 III) detected by a restriction enzyme mapping. This is found frequently in Melanesians and Polynesians but not in five other populations in which alpha-thalassemia is prevalent. We used restriction enzyme haplotype analysis to support a single origin for this mutation and propose that it is a useful population marker. Its geographical distribution supports a route through Island Melanesia for the colonizers of Polynesia. PMID- 2988336 TI - Superficial laser vulvectomy. II. The anatomic and biophysical principles permitting accurate control over the depth of dermal destruction with the carbon dioxide laser. AB - The rationale for using the carbon dioxide laser to treat either vulvar intraepithelial neoplasia or extensive papillomaviral infections is to destroy the entire area of abnormal epithelium to a shallow depth, so that rapid healing will occur from normal keratinocytes in the underlying pilosebaceous glands. After the first laser impact, anatomic landmarks in the crater base are disguised by a layer of charred proteins, and any structure that is visible will already have suffered thermal necrosis. Accurate control of depth depends upon special surgical strategies that correlate the level of the underlying zone of thermal necrosis with specific visual appearances within the zone of vaporization. Maneuvers that limit depth of penetration to one of three desirable surgical planes (basement membrane, papillary dermis, midreticular dermis) are described. PMID- 2988337 TI - Long-term survival rates with various chemotherapeutic regimens in stages III and IV ovarian adenocarcinoma. The influence of optimum pretreatment surgical resection. AB - This report compares long-term survival rates for patients treated with four different chemotherapeutic regimens for Stages III and IV ovarian adenocarcinoma. The patients were entered into consecutive, prospective, randomized studies with an essentially common chemotherapeutic arm. The first study compared the single agent melphalan with actinomycin D, 5-fluorouracil, and Cytoxan. The second study compared 5-fluorouracil plus Cytoxan and methotrexate-leucovorin rescue plus Cytoxan. The patient characteristics in the two studies were very similar except for more aggressive tumor-reductive operations in the second study. Observed survival rates for the first 2 years in the second study were very much higher than in the first study. However, by the third, fourth, and fifth years, the survival rates of the 5-fluorouracil-Cytoxan-treated individuals had reached the same low levels seen in the first study. It appears that an optimum surgical procedure by itself may enhance survival during the first 2 years. Survival with methotrexate-leucovorin rescue plus Cytoxan was statistically significantly better than with melphalan or actinomycin D-5-fluorouracil-Cytoxan. Third-, fourth-, and fifth-year survival rates with methotrexate-leucovorin rescue plus Cytoxan were substantially higher than with 5-fluorouracil-Cytoxan; however, the survival distributions for these two treatments were not statistically significantly different. Long-term survival rate data for patients with Stages III and IV ovarian adenocarcinomas treated with chemotherapy are rare. The 19% 5 year survival rate with methotrexate-leucovorin rescue plus Cytoxan in the present study is considerably higher than other reported survival rates. PMID- 2988338 TI - Effect of treatment with sodium valproate on plasma adrenocorticotropic hormone and cortisol concentrations in pregnancy. AB - To investigate whether treatment with valproic acid in pregnancy decreases adrenocorticotropic hormone and cortisol secretion, we measured their concentrations in maternal and umbilical cord blood from a patient receiving valproic acid. We found them to be normal. Valproic acid may not decrease adrenocorticotropic hormone/cortisol levels in pregnant patients or their fetuses. PMID- 2988339 TI - Sonographic appearance of the ovarian thecoma. PMID- 2988340 TI - Tympanoplasty with calcium phosphate. AB - A new approach to the problems of chronic middle ear disease using the hydroxyapatite prosthesis is described. Open surgery is preferred and, independent of the eradication, reconstruction is possible. The posterior canal wall is reconstructed with a canal wall prosthesis of porous hydroxyapatite. The middle ear chain defects are reconstructed with a prosthesis of dense hydroxyapatite. In the case of a cavity with a mobile bare footplate a total alloplastic middle ear of hydroxyapatite is used. PMID- 2988341 TI - Prolactin increases lipid fluidity and prolactin binding of rat prostatic membranes. AB - The objective of these studies was to determine whether prolactin could modify the lipid fluidity of rat ventral and dorsolateral prostate membranes and subsequently modify the availability of prolactin receptors. Additional studies were also undertaken to determine the effects of prolactin on serum lipid fluidity. Adult male rats were injected with 0, 100, or 400 micrograms ovine prolactin/day subcutaneously for a period of 5 days. Serum and prostatic membrane lipid fluidity was measured by a fluorescence polarization method using a lipid probe 1,6-diphenylhexatriene. Prolactin binding in dextran-coated charcoal pretreated prostatic membranes was determined by radioreceptor assay. This pretreatment has been reported by us to remove the endogenous substances that interfere with prolactin binding assay (J. R. Dave and R. J. Witorsch, Endocrinology 111: 2144-2146, 1982). Prolactin binding increased by approximately 44 and 72% in dorsolateral prostate and 16 and 39% in ventral prostate in 100- and 400-micrograms groups, respectively. Membrane fluidity increased by approximately 16 and 19% in dorsolateral prostate and 10 and 13% in ventral prostate in 100- and 400-micrograms groups, respectively. Serum lipid fluidity increased 50 and 79% in 100- and 400-micrograms groups, respectively. PMID- 2988342 TI - Intestinal metabolism and portal venous transport of 1,25(OH)2D3, 25(OH)D3, and vitamin D3 in the rat. AB - We compared the intestinal absorption of three vitamin D3 sterols and tested the hypothesis that the intestine hydroxylates absorbed vitamin D and transports polar metabolites in portal venous blood. Micellar solutions containing 50 nmol of a radiolabeled vitamin D3 sterol (1,25-dihydroxyvitamin D3, 25-hydroxyvitamin D3, or vitamin D3) were placed in closed jejunal segments of rats prepared with lymphatic and mesenteric venous fistulas. Venous blood loss was replaced by infusion of donor rat blood into the saphenous vein. After 1-2 h the rats were killed, and intestinal homogenates, mesenteric blood, and lymph were analyzed. The average rate of absorption of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] was two- and fivefold higher than that of 25-hydroxyvitamin D3 [25(OH)D3] and vitamin D3 (D3), respectively. Transport of hydroxylated vitamin D sterols was primarily via the venous route; the average rate of venous transport of 1,25(OH)2D3 was 18.3 X 10(2) nmol X min-1 X g-1 compared with 8.8 X 10(2) for 25(OH)D3 and 0.13 X 10(2) for D3. High-performance liquid chromatography of intestinal and plasma extracts revealed that there was 25-hydroxylation of absorbed D3, 24- and putative 1 hydroxylation of absorbed 25(OH)D3, and prompt portal venous transport of all hydroxylated metabolites. When 1,25(OH)2D3 was infused into the lumen, the composition of radiolabeled sterols found in intestinal homogenates and mesenteric venous plasma was virtually identical to that of the infusate. These studies provide in vivo evidence for the intestinal metabolism of pharmacological quantities of absorbed vitamin D3 sterols and the prompt portal venous transport of more polar metabolites. PMID- 2988343 TI - Preferential binding of vasoactive intestinal peptide to hepatic nonparenchymal cells. AB - The binding of 125I-vasoactive intestinal peptide (125I-VIP) and 125I-insulin has been examined in highly enriched populations of rat hepatocytes and hepatic nonparenchymal cells. 125I-VIP bound to high-affinity sites (Ka = 1.7 X 10(9) M 1) in nonparenchymal cells. Specific binding in these cells was nearly fivefold greater than in hepatocytes (15.0 +/- 0.6% vs. 3.6 +/- 0.7% radioactivity bound per 4 X 10(5) cells). In contrast, 125I-insulin binding was similar in both cell populations (18.2 +/- 2.4% per 4 X 10(5) cells in hepatocytes vs. 17.1 +/- 1.0% in nonparenchymal cells). Glucagon and insulin had no effect on 125I-VIP binding in nonparenchymal cells. Secretin inhibited 125I-VIP binding but was only about 1% as potent as unlabeled VIP. VIP had no apparent effect on cAMP levels in either cell population, whereas glucagon increased cAMP levels in both cell types. Our findings suggest that VIP binds preferentially to hepatic nonparenchymal cells and that these cells are primarily responsible for the clearance of VIP from the portal circulation. PMID- 2988345 TI - Kinin stimulation of renin release in isolated rat glomeruli. AB - Numerous studies have suggested that a functional relationship exists between the renal kallikrein-kinin and renin-angiotensin systems. We used an in vitro preparation of isolated glomeruli to study the effect of kinins on renin release. Glomeruli were harvested from the dissected cortices of rat kidneys by a passive sieving technique. They were then placed in chambers and superfused with aerated modified Krebs buffer, and the chamber effluent was collected during two sequential 10-min periods. Bradykinin at 10(-6) M increased renin release from 2.17 +/- 0.34 to 4.05 +/- 0.72 ng ANG I/min, and 10(-5) M bradykinin increased renin release from 3.51 +/- 0.64 to 8.94 +/- 1.27 ng ANG I/min. With albumin modified buffer, 10(-5) M bradykinin stimulated renin release from 7.11 +/- 0.79 to 14.03 +/- 2.36 ng/min. Lys-bradykinin at 10(-5) M also increased renin release from 5.69 +/- 1.11 to 14.50 +/- 2.65 ng ANG I/min. However, neither of the inactive kinin analogues, [Tyr8]bradykinin or des-Arg9-bradykinin, had any effect on renin release. We found that isolated glomeruli were relatively free of kininase activity, in contrast to the high activity found in the kidney slice preparation or in rat plasma. These results suggest that biologically active kinins can stimulate renin release in an in vitro preparation that is free of degradative kininases and independent of hemodynamic and neural influences. PMID- 2988344 TI - Effects and interrelationships of PTH, Ca2+, vitamin D, and Pi in acid-base homeostasis. AB - Chronic administration of 1,25(OH)2D or PTH increases the set point at which plasma bicarbonate concentration is regulated by the kidney and thereby maintains metabolic alkalosis in a variety of species. The renal mechanism(s) responsible for the chronic acid excretory response to 1,25(OH)2D and PTH administration have not been defined, but indirect evidence suggests effects on distal nephron segments. With either hormone in dogs, but not in rats, metabolic alkalosis is generated in part by extrarenal mechanisms. Contrary to the variable finding of hyperchloremic acidosis in human primary hyperparathyroidism, a steady state of mild metabolic alkalosis of renal origin is achieved in normal human subjects infused chronically with PTH. The multiple potential mechanisms responsible for this discrepancy will require careful consideration in future investigations. The acute and chronic effects of plasma calcium concentration on both renal and extrarenal acid-base homeostasis are incompletely understood and require extensive further investigation. Studies in rats have suggested that phosphate depletion results in important counterbalancing renal acidosis and extrarenal alkalosis-producing effects. Chronic hypophosphaturia in the absence of appreciable hypophosphatemia can also result in impaired renal acidification by virtue of phosphate's property as a luminal buffer. PMID- 2988346 TI - Alpha 2-adrenoceptor antagonism of vasopressin-induced changes in sodium excretion. AB - Alpha 2-adrenoceptor agonists attenuate vasopressin-mediated changes in water excretion. The effects on sodium excretion, however, are unclear. We therefore utilized the nonrecirculating isolated perfused rat kidney to study the direct effects of vasopressin and alpha 2-adrenoceptor stimulation on sodium and water excretion in the absence of systemic regulatory systems. The perfusate was a Krebs-Henseleit solution (3.5 g/100 ml Ficoll; 1.0% albumin; 36 degrees C) containing prazosin (30 nM) and propranolol (100 nM) to prevent effects of alpha 1- and beta-adrenoceptor stimulation. Vasopressin (10 microU/ml) produced a significant (P less than 0.05) decrease in both water and sodium excretion. Potassium excretion was not significantly altered. Alpha 2-Adrenoceptor stimulation with l-epinephrine (28 nM) reversed (P less than 0.05) the effects of vasopressin on water and sodium excretion. To confirm that this attenuation was mediated by alpha 2-adrenoceptors, an alpha 2-adrenoceptor antagonist, yohimbine, was administered. Yohimbine (300 nM) blocked the effects of epinephrine on sodium and water excretion (P less than 0.05). The adenosine P-site agonist, SQ 22,536 (100 microM), which mediates its effects through inhibition of adenylate cyclase, produced the same reversal as that of epinephrine on vasopressin-mediated changes. Thus alpha 2-adrenoceptor stimulation antagonized the effects of vasopressin on both water and sodium excretion at the renal level. A corollary to this conclusion is that the function-specific activation of renal adenylate cyclase determines the effect of alpha 2-adrenoceptor stimulation. PMID- 2988347 TI - Amiloride inhibition of proximal tubular acidification. AB - In brush border membrane vesicles prepared from mammalian kidney cortex, amiloride is a potent inhibitor of the Na+/H+ exchanger. In the present study, in vivo microperfusion was used to examine the effect of luminal amiloride on transport in the rat superficial proximal convoluted tubule. At a perfusion rate of 14 nl/min, addition of 10(-3) M amiloride to artificial early proximal tubular fluid reduced bicarbonate absorption from 103 +/- 7 to 81 +/- 5 pmol mm-1 X min-1 and volume absorption from 2.03 +/- 0.15 to 1.57 +/- 0.06 nl X mm-1 X min-1. Glucose efflux was unchanged, excluding nonspecific inhibition of Na+-K+-ATPase. Luminal amiloride at 10(-4) M did not affect bicarbonate absorption or volume absorption. At a perfusion rate of 41 nl/min, 10(-3) M amiloride reduced bicarbonate absorption from 179 +/- 8 to 114 +/- 9 pmol X mm-1 X min-1, a significantly greater inhibition than that seen in tubules perfused at 14 nl/min. Amiloride at 10(-3) M had no significant effect on sodium chloride absorption as measured by volume flux from an artificial late proximal tubular fluid. The results show that luminal amiloride specifically inhibits proximal acidification and demonstrate involvement of the Na+/H+ antiporter in proximal tubular acidification. However, the inhibition of acidification is less than the inhibition of Na+/H+ exchange predicted by vesicle studies. PMID- 2988348 TI - Role of citrate excretion in acid-base balance in diuretic-induced alkalosis in the rat. AB - Studies were performed to assess the role of changes in the excretion of citrate, a metabolic precursor of bicarbonate, in acid-base balance in diuretic-induced metabolic alkalosis. Rats on a low-chloride diet with sodium sulfate added were studied during a base-line period, 3 days of furosemide administration, and 4 days post-furosemide. During the period of furosemide administration, net acid excretion and plasma bicarbonate concentration increased. In the post-furosemide period, net acid excretion remained higher than base line but plasma bicarbonate concentration did not increase further. Citrate excretion was significantly higher in the post-furosemide period than in base line. Studies substituting sodium neutral phosphate or sodium bicarbonate for dietary sodium sulfate demonstrated greater increases in net acid excretion post-furosemide and, again, no increase in plasma bicarbonate concentration during this period. Citrate excretion was greater than in the sulfate group. The increment in citrate excretion was proportional to the base "load," defined with respect to changes in net acid excretion and/or dietary bicarbonate. Thus, in these studies alterations of base excretion in the form of citrate play an important role in acid-base balance during diuretic-induced metabolic alkalosis. PMID- 2988349 TI - Sodium-dependent bicarbonate absorption by cortical thick ascending limb of rat kidney. AB - In vitro microperfusion experiments were performed to investigate the mechanism of bicarbonate absorption in the cortical thick ascending limb of the rat. Tubules were perfused at 1.0-1.5 nl X min-1 X mm-1 and bicarbonate concentration was 25 mM in the perfusate and bath. Bicarbonate absorption rates were determined by microcalorimetry. Control tubules absorbed bicarbonate at a mean rate of 9.5 +/- 0.6 pmol X min-1 X mm-1. The limiting luminal bicarbonate concentration was approximately 5 mM for tubules perfused at slow rates with 25 mM bicarbonate in the bath. Acetazolamide (10(-4)M) in the bath reduced bicarbonate absorption by 76% without significant effect on transepithelial voltage. Removing sodium from the perfusate and bath or removing potassium from the bath reduced bicarbonate absorption and transepithelial voltage to near zero. Adding amiloride (5 X 10(-4) or 10(-3) M) to the perfusate reduced bicarbonate absorption by 60-75% without detectable effect on transepithelial voltage. Adding furosemide (10(-4)M) to the perfusate increased bicarbonate absorption significantly by 40-50% while decreasing transepithelial voltage from 17 to 1.8 mV. Thus, bicarbonate absorption by cortical thick ascending limbs requires carbonic anhydrase activity and sodium transport but is not dependent on transepithelial voltage. When considered together, the results are consistent with mediation of the bicarbonate absorption by apical membrane sodium-hydrogen exchange. PMID- 2988350 TI - Cephalic carotid pressure as a measure of transmission through cervical ganglion. AB - Cephalic or cranial pressure, i.e., pressure recorded craniad from the external carotid artery, was recorded in the cat during stimulation of the pre- and postganglionic trunks of the superior cervical ganglion (SCG) as was tension developed by the nictitating membrane under these situations. The influence of distension of the carotid sinus and of metoprolol on hemodynamic and tension responses was compared with control responses. Distension of the sinus with an indwelling balloon produced an increase of cephalic pressure evoked by stimulation of the SCG. Tension developed by the nictitating membrane was unaffected. Metoprolol reduced the increase in pressure resulting from stimulation of the preganglionic trunk of the SCG at 1.0 Hz but enhanced the pressure increase in the cranial circulation evoked by postganglionic stimulation of the SCG at 5.0 Hz. Action potentials, recorded postganglionically from the SCG, could be roughly grouped by height and latency into three populations. The height of the M3 population increased with distension of the carotid sinus. This study provides evidence that cephalic pressure can be used as a measure of the influence of various factors on the transmission of impulses through the SCG. This parameter appears to be a more sensitive measure of transmission than that of tension developed by the nictitating membrane. Distension of the carotid sinus is one factor that facilitates ganglionic transmission, whereas the beta adrenoceptor antagonist metoprolol depresses it. PMID- 2988351 TI - Isolation and functional characterization of crustacean larval salt gland. AB - A batch method for isolating viable salt glands from the naupliar brine shrimp (Artemia salina) has been developed. This protocol produces a final preparation consisting of approximately 185 isolated salt glands, representing 1 X 10(4) secretory cells/g wet wt nauplii, with a final purity of 88%. Assays of cell integrity and function indicate good retention of in situ characteristics. Vital dye was excluded by 95% of the cells for at least 24 h. The O2 consumption rate was 22.7 nM O2 X min-1 X mg protein-1 and could be altered predictably by compounds known to affect oxidative phosphorylation and ion transport. The specific activity of the Na+-K+-ATPase in the salt gland, measured here for the first time, was 9.1 mM Pi X h-1 X mg protein-1. This is a substantial proportion of the body total, 17%, as expected for an active ion-transporting epithelium. PMID- 2988352 TI - Serologic markers of hepatitis A and B in the population of Bali, Indonesia. AB - A total of 343 sera from Balinese subjects in different age groups and geographic locations were tested by radioimmunoassay (RIA) for serum antibodies to hepatitis B surface antigen (anti-HBs) and hepatitis B core antigen (anti-HBc); most sera were also tested for hepatitis B surface antigen (HBsAg), and for antibody to hepatitis A virus (anti-HAV). One hundred percent of the adult population was found to have anti-HAV, with antibody acquisition beginning in early childhood and reaching a level of 95% by the age of 10 years. Antibodies to hepatitis B virus were also frequent in young children, rapidly peaking to near 80% in older children and adolescents, then declining to a plateau that fluctuated between 40% and 60% throughout adult life. Overall, anti-HBc (49%) was detected slightly more often than anti-HBs (45%), but the relative frequencies of the 2 antibodies varied considerably from group to group. Despite these high antibody prevalences, HBsAg was detected in only 1.5% of the general population, and in no woman of child-bearing age. In utero infection is thus far less likely to account for the early acquisition of antibody to hepatitis B virus than inapparent percutaneous transmission occurring under conditions of close personal contact. PMID- 2988354 TI - Suicide attempted by self-immolation during adolescence. II. Psychiatric treatment and outcome. PMID- 2988353 TI - Prevalence of serological hepatitis A and B markers in a rural area of northern Zaire. AB - In a seroepidemiological study of the population of a rural area in northern Zaire, markers of hepatitis A and B were determined. Examinations of serum specimens (n = 142) showed that hepatitis A is acquired early in childhood; virtually all persons beyond 20 years of age are immune. Capillary blood dried on filter discs (n = 352) gave accurate results only in childhood; in older individuals a high percentage of positive results was missed (approximately 20%), presumably due to lower anti-HAV titers in older persons. The dried-blood method showed similar limitations regarding the detection of hepatitis B markers. In the 5-7 year age group there was already a high prevalence of anti-HBc (59%) as determined by the dried blood method. In the 10-19 year age group the prevalence was 94% as determined by examination of serum specimens. In all age groups the percentage of HBsAg positive persons was 20.7%, as demonstrated in capillary blood specimens and 31.7% in serum specimens. PMID- 2988355 TI - [Congenital infections due to Coxsackie viruses]. PMID- 2988356 TI - Natural history, diagnosis, and management of Epstein-Barr virus infection. PMID- 2988357 TI - Viral infections of the liver. PMID- 2988358 TI - Cytomegalovirus infection. PMID- 2988359 TI - Therapy for human herpesvirus infections. A perspective. PMID- 2988360 TI - The effect of ACTH on cerebral blood flow in children with intractable epilepsy. AB - Cerebral blood flow was assessed by ultrasound in 12 children with intractable epilepsy who were treated with ACTH. The average maximal blood velocity (A/L) and end-diastolic blood velocity (d) of the internal carotid artery were measured, before, during and after ACTH therapy in each subject. The right and left mean A/L and d values were significantly decreased during ACTH therapy, and these values returned to the previous levels after the treatment. Cerebral function in children treated with ACTH may be affected by a decrease in cerebral blood flow. PMID- 2988361 TI - Malignant hyperthermia. PMID- 2988362 TI - What's new in neurosurgery in the 1980's. PMID- 2988363 TI - Ethanol and membrane lipids. AB - Although ethanol is known to exert its primary mode of action on the central nervous system, the exact molecular interaction underlying the behavioral and physiological manifestations of alcohol intoxication has not been elucidated. Chronic ethanol administration results in changes in organ functions. These changes are reflective of the adaptive mechanisms in response to the acute effects of ethanol. Biophysical studies have shown that ethanol in vitro disorders the membrane and perturbs the fine structural arrangement of the membrane lipids. In the chronic state, these membranes develop resistance to the disordering effects. Tolerance development is also accompanied by biochemical changes. Although ethanol-induced changes in membrane lipids have been implicated in both biophysical and biochemical studies, measurements of membrane lipids, such as cholesterol content, fatty acid unsaturation, phospholipid distribution, and ganglioside profiles, have not produced conclusive evidence that any of these parameters are directly involved in the action of ethanol. On the other hand, there is increasing evidence indicating that although ethanol in vitro produces a membrane-fluidizing effect, the chronic response to this effect is not to change the membrane bulk lipid composition. Instead, changes in membrane lipids may pertain to small metabolically active pools located in certain subcellular fractions. Most likely, these lipids are involved in important membrane functions. For example, the increase in PS in brain plasma membranes may provide an explanation for the adaptive increase in synaptic membrane ion transport activity, especially (Na,K)-ATPase. There is also evidence that the lipid pool involved in the deacylation-reacylation mechanism (i.e., PI and PC with 20:4 groups) is altered after ethanol administration. An increase in metabolic turnover of these phospholipid pools may have important implications for the membrane functional changes. Obviously, there are other lipid-metabolizing enzyme systems that may exert similar effects but have not yet been investigated in detail. From the results of these studies, it is concluded that the multiple actions of ethanol are associated with changes in enzymic systems important in the functional expression of the membranes. PMID- 2988365 TI - A reflectance method for quantification of immunological reactions on surfaces. AB - A simple optical method for determination of thicknesses of organic layers on solid substrates is described. The method is based on the use of a substrate with a high refractive index, e.g., silicon, and reflection of light at an angle of incidence close to the pseudo-Brewster angle. Under these conditions, a reflectance minimum is obtained for light polarized in the plane of incidence. The presence of an organic layer on the surface will increase the reflectance, which is used to determine the thickness of the layer. The physical basis of the method is described briefly, and the application to immunology is demonstrated. PMID- 2988364 TI - Ethanol oxidation by hydroxyl radicals: role of iron chelates, superoxide, and hydrogen peroxide. AB - Oxygen-derived free radicals such as the hydroxyl radical (.OH) have been shown to mediate the oxidation of ethanol by a variety of oxy radical-generating systems. Among these are microsomal electron transport systems (both intact and purified, reconstituted systems), the coupled oxidation of hypoxanthine or xanthine by xanthine oxidase, and the model iron-ascorbate system. The sequence of reactions leading to the oxy radical-dependent oxidation of ethanol as well as other hydroxyl radical-scavenging agents by these various systems is believed to proceed through the formation of a common intermediate, namely, hydrogen peroxide (H2O2), after dismutation of the superoxide anion radical (O2-.). The presence of iron, especially chelated iron, greatly enhances the production of .OH by serving as an oxidant for O2-. or a reductant for H2O2. Experiments were carried out to evaluate the role of iron, the chelating agent, O2-., and H2O2 in the oxidation of ethanol by a variety of in vitro systems (chemical, enzymatic, and intact membrane bound) that can produce oxy radicals via different mechanisms. The generation of .OH by all the systems studied was sensitive to catalase, which indicates that H2O2 is the precursor of .OH. Superoxide radical appears to be the reducing agent in the hypoxanthine-xanthine oxidase system, indicating an iron catalyzed Haber-Weiss reaction. In the ascorbate, reductase, and microsomal systems, superoxide radical does not appear to be the reducing agent. However, superoxide radical probably is the precursor of H2O2. While iron plays an important role in the production of .OH by the various systems, the effect of iron depends on the nature of the iron chelate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988366 TI - Carbon nuclear magnetic resonance spectra of oligosaccharides isolated from human milk and ovarian cyst mucin. AB - Natural abundance Carbon-13 nuclear magnetic resonance spectra at 20 MHz were reported for the three common human milk oligosaccharides, lacto-N-tetraose and lacto-N-fucopentaoses I and II, as well as for two related tetra- and hexasaccharide alditols isolated from the alkaline borohydride degradation products of an ovarian cyst glycoprotein. Spectral assignments made with the help of deuterium-induced shift (DIS), attached proton test (APT), and T1 data indicated some very irregular glycosylation shifts which were attributed to effects of steric crowding and non-nearest-neighbor interactions. Samples as small as 10 mumol of oligosaccharide gave acceptable 20-MHz spectra with the use of a 5-mm probe coil. PMID- 2988367 TI - An assay method for cyclic AMP using high-performance liquid chromatography with pretreatment by alkaline phosphatase. AB - A convenient method for determination of cyclic AMP is described. This nucleotide was determined using high-performance liquid chromatography after the treatment of tissue extracts by alkaline phosphatase. Tissue extracts contain large amounts of materials which interfere with the measurement of cyclic AMP on the chromatogram. By the alkaline phosphatase treatment, these materials were completely converted to compounds which no longer interfered with the measurement. This method enables the detection of 2 pmol cyclic AMP, and is applicable to various tissue extracts which contain at least 0.1 nmol cyclic AMP/g wet wt. PMID- 2988369 TI - A new competition assay for the solubilized hepatic galactosyl receptor. AB - The present method of quantitating soluble asialoglycoprotein (galactosyl) receptor activity relies on the selective precipitation of receptor-ligand complexes to allow separation from free ligand. To provide an alternative to selective precipitation procedures, a simple and rapid method to assay for detergent-solubilized galactosyl receptor activity has been developed which uses permeabilized, fixed cells as a source of immobilized solid-phase receptors. Isolated rat hepatocytes were treated with digitonin to make available the internal as well as the external receptors. The permeable cells were also treated with glutaraldehyde to prevent further protein loss during subsequent exposure to detergents such as Triton X-100. The permeable/fixed cells, which retained about 70% of their total 125I-asialo-orosomucoid (125I-ASOR)-binding activity, with 89% specific binding, were insoluble even in 0.5% Triton X-100 and were easily pelleted. The permeable/fixed cells can be prepared in advance and stored frozen for months. A detergent extract of receptor is mixed with a constant amount of both 125I-ASOR and permeable/fixed cells. Soluble active receptors compete with immobilized receptors on the treated cell for binding of the 125I-ASOR. The assay is reproducible, linear over a broad range of soluble receptor concentration, and can quantitate receptor activity from as few as 10(5) hepatocytes. A modified purification procedure for the rat hepatic galactosyl receptor using this competition assay is also described. PMID- 2988368 TI - Assay of hybrid ribonuclease using a membrane filter-immobilized synthetic hybrid: application to the human leukemic cell. AB - A method for assaying hybrid ribonuclease has been devised which utilizes as substrate the synthetic hybrid [3H]polyriboadenylic acid [poly(rA)]:polydeoxythymidylic acid [poly(dT)] immobilized on the solid matrix of nitrocellulose filters. The hybridization on filter of [3H]poly(rA) to poly(dT) has been explored in terms of efficacy of the process and the response of the product to RNase H. A pulse of uv irradiation of poly(dT) while in dry state on the filter increased its firm binding to the filter in a concentration-dependent manner, resulting in a concomitant increase of the yield of hybrid formation. The filter-immobilized hybrid was 95% resistant to RNase A but sensitive to RNase H. When stored in toluene in the cold the hybrid maintained its stability for over 6 months, as judged by its resistance to RNase A. The method offers a number of advantages over assays that use solution hybrids as substrates and was readily applicable in the screening of leukemic patients, in the leukocytes of which it has demonstrated increased RNase H levels. PMID- 2988370 TI - High-performance anion-exchange chromatography of oligonucleotides. AB - Several types of high-performance silica-based supports have been found to be effective in the separation of polynucleotides. The principal difference in these materials is the type of bonded phase and the method by which it is attached to the silica support. One approach is the coupling of stationary-phase groups to the surface through siloxane bonding. This technique is simple and produces a material of high capacity and resolution, but it suffers from poor bonded-phase stability. An alternative approach is the adsorption of low-molecular-weight polyethylene imines (PEI) that are crosslinked into a surface film. The stationary phase is held in place by adsorption of the film at multiple sites. A previous report on this material showed the resolution of oligonucleotides containing up to 30 bases. This paper reports further optimization of the PEI bonding chemistry in the preparation of HP-IEC columns for oligonucleotides and tRNA species. Quaternization of the ion-exchange matrix was found to increase resolution of oligonucleotides from 30 to 50 bases. The same support was found to be capable of resolving multiple tRNA species. Separations were achieved on small (0.42 X 5 cm) columns, using a 60- to 120-min ammonium sulfate gradient. The initial solvent was 15% acetonitrile in 0.05 M potassium phosphate (pH 5.9). The addition of 1 M ammonium sulfate to the initial solvent was used to prepare the final solvent. PMID- 2988371 TI - A combination of methods for the preparation of highly intact mitoplasts from beef heart mitochondria. AB - Beef heart mitochondria are noted for difficulty of fractionation compared to mitochondria from other tissues. A method that would give the desired efficiency while leaving the inner membrane intact did not seem to be available. Four variables, alone and in all possible combinations, were evaluated for their effectiveness in fractionation. These variables were treatment with digitonin, hypotonic swelling, phospholipase A2 predigestion, and addition of high concentrations of EDTA. The efficiency of fractionation was evaluated by electron microscopy and marker enzyme assays. Highly intact mitoplasts were obtained when the mitochondria were treated with digitonin together with intense hypotonic swelling, or moderate hypotonic swelling and phospholipase A2 predigestion, or EDTA addition and phospholipase predigestion. The enzyme GTP:AMP phosphotransferase was found to be located exclusively in the matrix compartment. PMID- 2988373 TI - Restriction site polymorphisms in the pig beta-globin gene cluster. AB - A restriction fragment length polymorphism was detected in pig DNA digested with Hind III restriction endonuclease and probed with rabbit beta 1-globin gene. Eight different phenotypes were observed and for six of them family data demonstrated that they are determined by three alleles. As this polymorphism is not found with four other restriction endonucleases (Bam HI, Eco RI, Kpn I, and Pst I), single point mutations are proposed to explain the observed differences. PMID- 2988372 TI - [Changes in renal blood flow and uropoiesis during alpha-adrenoreceptor blockade in shock]. PMID- 2988374 TI - Evidence for mapping pig C4 gene(s) within the pig major histocompatibility complex (SLA). PMID- 2988375 TI - Pathologic effects of intrauterine deposition of pseudorabies virus on the reproductive tract of swine in early pregnancy. AB - Pseudorabies virus was inoculated into the uterus of 15 gilts within 6 hours after natural breeding, and gilts were necropsied at postbreeding days (PBD) 3, 6, 10, 14, and 28; 3 control gilts were treated similarly, except for inoculation with pseudorabies virus and were necropsied at PBD 6, 10, and 14. Tissues were collected for virus isolation, fluorescent antibody staining, and histopathologic examination. Pseudorabies virus was isolated from the reproductive tract up to day 14. Lesions in the reproductive tract consisted of multifocal to diffuse lymphohistiocytic vaginitis and endometritis, and lymphoplasmacytic aggregates in the corpora lutea. Multiple ulcers were seen in the vagina or endometrium of several gilts at PBD 3, 6, and 10. Corpora lutea of 1 gilt were necrotic at PBD 14 and contained large numbers of inflammatory cells. Focal aggregates of lymphocytes and plasma cells were seen in vagina and endometrium of 3 gilts and in the ovary of 1 gilt at day 28. PMID- 2988376 TI - Immunologic response of sheep to inactivated and virulent bluetongue virus. AB - Humoral and cellular immune responses of sheep to inactivated and virulent bluetongue virus (BTV) were studied. All sheep inoculated with inactivated BTV developed BTV group-specific nonneutralizing antibodies, as determined by agar gel immunodiffusion. The development of group-specific, nonneutralizing, complement-fixing antibodies was variable and appeared to be dependent on immunizing BTV serotype, sheep breed, and individual variation. Virus neutralizing antibodies were never detected after inoculation with the inactivated BTV. In vitro lymphocyte stimulation to BTV soluble antigen was observed with cells from all inoculated Warhill sheep and with cells from 1 of 3 inoculated Suffolk cross sheep. Complement-fixation titers did not appear to correlate with the degree of protection observed, ie, duration of postchallenge exposure viremia. The development of postchallenge-exposure neutralizing antibody titer was inversely correlated to protective immunity. The development of a response to BTV antigen in the lymphocyte-stimulation test associated most closely with protection. Warhill sheep were afforded better protection, by inoculation with inactivated BTV, to live virus challenge exposure than were the Suffolk cross sheep. Approximately 30% of the inoculated Suffolk cross sheep responded to challenge exposure with intensified clinical signs of blue-tongue, compared with the challenge-exposed control sheep of the same breed. PMID- 2988377 TI - Retroviral-associated eosinophilic leukemia in the cat. AB - Fourteen specific-pathogen-free cats were inoculated with a putative env gene recombinant feline retrovirus, PR8. An isolate of the Rickard strain of feline leukemia virus (FeLV-R), PR8, has the properties of both an exogenous (FeLV-R) and an endogenous (xenotropic) feline retrovirus (RD-114). Twelve of the PR8 inoculated cats developed viremia; 2 of the 12 cats developed eosinophilia, with 1 being diagnosed with eosinophilic leukemia and the other with extreme eosinophilic hyperplasia. Eosinophilic leukemia is rare in cats and has not previously been associated with retroviral infection. Changes in the viral envelope properties may have altered the pathogenicity of the exogenous virus to cause this rare form of leukemia. PMID- 2988378 TI - Infection of early bovine embryos with bovine herpesvirus-1. AB - Recently hatched bovine embryos were exposed in vitro to 1 of 4 strains of bovine herpesvirus-1 to determine whether the viruses would replicate in these embryos and, if so, what pathologic consequences would ensue. Exposure to each of the viruses resulted in embryonic infection and death, and replication of the agents was demonstrated by electron microscopy and titration of progeny virus. There were no dramatic differences between virus strains in pathogenicity or in the ultrastructural pathologic findings of infection. PMID- 2988379 TI - Prospective study of progeny of inapparent equine carriers of equine infectious anemia virus. AB - Progeny of a band of horses, positive by the agar-gel immunodiffusion (AGID) test for equine infectious anemia (EIA) antibody, were observed through their weaning over a 4-year period. Sentinels (AGID test-negative) were allowed to mingle with EIA-infected mares and their foals in pasture situations in an area with high populations of potential vectors. Of 27 adult sentinels, 8 (30%) seroconverted in annual rates ranging from 0% to 75%. In contrast, only 2 of 31 (6%) foals weaned became infected. Difference in infection rates between adult sentinels and foals was significant (chi 2, P less than 0.05). Possible explanations for differences included protective value of colostral immunity and differences in attractiveness to blood feeding vectors. Detectable colostral immunity to EIA virus in the AGID test persisted for 25 to 195 days, with a mean of 124 days. PMID- 2988380 TI - Pathologic changes caused by transplacental infection with an adenovirus-like agent in pigs. AB - Five hysterectomy-derived colostrum-deprived pigs housed in individual cages with positive ventilation developed severe skin cyanosis and edema of the subcutaneous tissues in the submandibular, thoracic, and abdominal regions. The 5 pigs were killed 7 to 10 days after birth. Eosinophilic intranuclear inclusion bodies were found in endothelial cells of capillary and small blood vessels throughout the body. Ultrastructurally, nuclei of these affected endothelial cells contained small crystalline arrays of virus particles, which were considered to belong to the adenovirus-like group from their size and structure. The present results indicated that the porcine adenovirus-like agent might also have the ability to produce transplacental infection. PMID- 2988381 TI - Transmission of the white-tailed deer cutaneous fibroma. AB - Cutaneous fibromas were successfully transmitted to 7 white-tailed deer (Odocoileus virginianus) inoculated with crude fibroma extracts (2 deer) or with partially purified deer fibroma virus (5 deer). The fibromas were transmitted by intradermal and subcutaneous inoculation and by rubbing the virus preparation into tattoo sites. Inoculation by scarification was not successful. The induced tumors resembled those of naturally occurring fibromas. Tattoo inoculation sites underwent an initial acute inflammatory response followed by mesenchymal proliferation, perivascular lymphocytic infiltration, and finally regression. The deer developed antibody titers against deer fibroma virus as determined by hemagglutination inhibition, using mouse RBC. Viral antigens could not be detected by indirect immunofluorescence in any induced fibroma. PMID- 2988382 TI - Ultrastructure of the intraerythrocytic stage of Cytauxzoon felis. AB - The erythrocytes of 2 cats experimentally infected with Cytauxzoon felis were examined by light and electron microscopy. In stained blood smears, parasitized erythrocytes usually contained a single, roundish organism, but occasionally up to 4 were present in a cell. Chains of these roundish organisms also were seen. Elongated parasites, sometimes with ear-like projections, were present in a few erythrocytes. By electron microscopy, the parasite contained a poorly defined nucleus, rough endoplasmic reticulum, ribosomes, nonplicated mitochondria, food vacuoles, and a cytostome on its limiting membrane. Usually, the parasite was oval, but budding forms also were evident. Crystalloid inclusions were present in parasitized and nonparasitized erythrocytes. PMID- 2988383 TI - Ornithodoros coriaceus (pajaroello tick) as a vector of bluetongue virus. AB - Preliminary studies demonstrated that the argasid tick, Ornithodoros coriaceus Koch, could become infected with bluetongue virus (BTV). Ticks became infected after feeding through artificial membranes on BTV-infected suspensions of cell cultures, chicken embryos, and sheep blood. Ticks also became infected after natural feeding on viremic sheep (BTV serotype 17) and cattle (BTV serotype 11). Virus was recovered from the hemolymph and salivary glands of ticks which had ingested BTV either through an artificial membrane or by natural feeding on a host animal. Ticks infected with BTV serotype 13 were capable of transmitting the virus to a susceptible cow at 42 days after ingestion of virus-infected cultures, thus demonstrating the potential of the tick to serve as a biological vector of BTV. PMID- 2988384 TI - Heterogeneity of immunologic function among subfractions of normal rat alveolar macrophages. II. Activation as a determinant of functional activity. AB - Previous studies have demonstrated differences in immunologic function among density-fractionated alveolar macrophages. The present study was undertaken to correlate these functional differences among alveolar macrophage density fractions with parameters of macrophage activation. Alveolar macrophages were lavaged from normal rats and separated into 5 density fractions by density gradient centrifugation. Cells from each density fraction were analyzed for parameters of macrophage activation: in vitro cytotoxic function directed against cultured neoplastic cells, ectoenzyme activities, and surface expression of Ia like determinants. Lavaged alveolar macrophages with in vitro cytotoxic function were concentrated among cells from higher density fractions, and cytotoxic function among the density fractions correlated inversely with ectoenzyme activities. Although the percentage of Ia-positive cells varied among the density fractions, surface expression of Ia-like determinants did not correlate positively with in vitro cytotoxic function. The results show that there is a variable distribution of cytotoxic function among alveolar macrophage density fractions that correlates with enzymatic markers of activation. Thus, we conclude that differential macrophage activation is a potential determinant of the functional heterogeneity observed in rat alveolar macrophages. PMID- 2988385 TI - Collagenase in the lower respiratory tract of patients with adult respiratory distress syndrome. AB - Collagenase activity in the bronchoalveolar lavage (BAL) of patients with adult respiratory distress syndrome (ARDS) was measured against Type I collagen (17 patients) and against Type III collagen (13 patients). Serine protease activity was also measured against Type III collagen (13 patients). Type I collagenase activity was detectable in 12 of 17 and Type III collagenase was detectable in 12 of 13 patients with ARDS. The 10 control subjects had no detectable Types I or III collagenase activity. Total and differential white cell counts were analyzed in the lavage fluid. Although the total counts did not differ between patients with ARDS and control subjects, the percentage of neutrophils was increased more than 25-fold and the percentage of macrophages was reduced almost 10-fold in the ARDS patients. Serial collagenase activity was followed in 1 ARDS survivor. In this patient Type III collagenase activity peaked before the Type I collagenase activity or serine protease activity reached their maximums. Both the latter enzyme activities paralleled the total recoverable cells in the BAL. PMID- 2988386 TI - Evidence for pancreas injury in adult respiratory distress syndrome. AB - The purpose of this study was to investigate the relation between serum lipase (LP), serum immunoreactive trypsin (IRT), and its inhibitors in patients with adult respiratory distress syndrome (ARDS) of diverse origin and to compare their time course with other acute conditions. The IRT and LP levels were determined at regular intervals in 41 patients hospitalized in the intensive respiratory unit with ARDS (n = 9), acute pancreatitis (n = 5), shock (n = 9), bronchopneumonia (n = 10), or acute cardiogenic pulmonary edema (n = 8). Several trypsin inhibitors were measured simultaneously: serum trypsin inhibitory capacity (TIC), alpha 1 antitrypsin, alpha 2-macroglobulin, and antithrombin III. Concomitantly, angiotensin-converting enzyme (ACE) activity was determined as a potential marker of the endothelial injury. A respective 19- and 13-fold increase in IRT and LP values were observed in patients with ARDS after a mean evolution of 6 days; similar increases were seen in patients with pancreatitis. These values were significantly higher than those observed in the other conditions studied. In patients with ARDS and acute pancreatitis, the evolution of IRT and LP values were associated with a sixfold rise in TIC. A low TIC/IRT ratio in patients with ARDS appeared to be an index of poor prognosis. Conversely, ACE activity evolution was characterized by an early decrease in all the conditions studied. These observations indicate that there is an acute delayed pancreas injury in ARDS. Thus, the release of pancreatic enzymes are not reliable markers of the early evolution of the disease but they may represent secondary mediators for enhancement of the increased endothelial permeability. PMID- 2988387 TI - Lymphoid interstitial pneumonitis in acquired immunodeficiency syndrome-related complex. AB - Three patients had a typical clinical, functional, and histologic pattern of lymphoid interstitial pneumonitis (LIP). All 3 also had a marked follicular and interfollicular hyperplasia of peripheral lymph nodes. A benign lymphocytic infiltration was seen in the kidneys in 2 patients, in the liver of 1 of these, and in the nasopharynx and the bone marrow in another. This LIP, associated with other diffuse benign lymphocytic infiltrates, could be part of AIDS-related manifestations. The patients originated from central Africa (2 patients) and Haiti (1 patient) where AIDS is endemic. They had several immune abnormalities frequently observed in AIDS such as a polyclonal hyperimmunoglobulinemia and, in the 2 patients studied, a decreased number of T4 lymphocytes and an inverted T4/T8 blood lymphocyte ratio. All 3 patients had serum antibodies to human T-cell leukemia viruses. PMID- 2988388 TI - Sjogren's syndrome and infectious mononucleosis. PMID- 2988389 TI - Exposure to human T-lymphotropic virus type III/lymphadenopathy-associated virus and immunologic abnormalities in asymptomatic homosexual men. AB - Immunologic and serologic studies were done in 120 homosexual men who reportedly had no symptoms related to the acquired immunodeficiency syndrome. Forty-nine men (41%) had antibody to the retrovirus human T-lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV), and 37 (31%) had an abnormal T-cell subset ratio and other immunologic abnormalities. These abnormalities were almost exclusively confined to men seropositive for the retrovirus. Multivariate statistical analysis showed that exposure to HTLV-III/LAV was the single best predictor of T-cell (and other) immunologic abnormalities. Sexual practice, particularly receptive anal intercourse, predicted exposure to HTLV-III/LAV but was not independently related to T-cell abnormalities. The association of other microbial serologic findings with HTLV-III/LAV seropositivity or T-cell changes was a coincidental function of sexual activity. Immunologic abnormalities in asymptomatic homosexual men are mostly related to exposure to HTLV-III/LAV. PMID- 2988390 TI - Bone disease in alcohol abuse. AB - We evaluated bone disease in eight white men between the ages of 49 and 61 years who had been abusing alcohol for at least 10 years. The mean density of vertebral cancellous bone was 58% of normal, whereas the mean density of appendicular cortical bone was 90% of normal. Marked reduction in active bone resorption and bone formation was seen without evidence of osteomalacia. Serum levels of calcium and magnesium were in the lower range of normal; serum levels of phosphorus, calcifediol, and calcitriol were normal; and serum levels of parathyroid hormone and nephrogenous cyclic adenosine monophosphate were in the higher range of normal. These data suggest that bone disease in these subjects is not due to inhibition of parathyroid hormone secretion or function, or abnormal vitamin D metabolism, but to an inhibition of bone remodeling by a mechanism independent of the calciotropic hormones. PMID- 2988391 TI - Treatment of hypercalcemia in parathyroid cancer with WR-2721, S-2-(3 aminopropylamino)ethyl-phosphorothioic acid. AB - The chemoprotective and hypocalcemic agent WR-2721, S-2-(3-aminopropylamino) ethyl-phosphorothioic acid, inhibits parathyroid hormone secretion in vivo and in vitro. We report the first clinical use of WR-2721 in refractory hypercalcemia secondary to parathyroid cancer. After several days of saline diuresis the patient received WR-2721, 740 mg/m2 over 15 minutes, resulting in a fall in serum calcium from 11.76 to 9.06 mg/dL within 24 hours. Serum parathyroid hormone levels decreased from 675 to 140 microLeq/mL 2 hours after the infusion was complete. When hypercalcemia recurred the patient was retreated with differing doses and infusion rates to determine the optimal method of drug administration to provide a satisfactory hypocalcemic response without adverse effects. In this patient, WR-2721 in intravenous boluses of 150 mg/m2 was effective without adverse effects. Using high-pressure liquid chromatography with electrochemical detection, plasma pharmacokinetic studies showed that WR-2721's distribution half life is 0.55 minutes. PMID- 2988392 TI - Cytomegalovirus infection and abnormal T-lymphocyte subset ratios in homosexual men. AB - T-helper:T-suppressor cell ratios of 1 or less were found in 2 of 42 homosexual men without antibodies to cytomegalovirus but in 33 of 67 homosexual men with antibodies (p less than 0.001). Of 34 men without antibody who were followed prospectively, 12 became seropositive for cytomegalovirus and all 12 developed helper: suppressor ratios of less than 1.0. These ratios remained at 1 or less for an average of 9.6 months but persisted for 15 months or more in 3 men. None of the men in the prospective study developed antibodies to the acquired immunodeficiency-syndrome-associated retrovirus. These results indicate that in the homosexual men studied, abnormally low T-lymphocyte helper: suppressor ratios occurred almost exclusively in those who were infected with cytomegalovirus, and in those prospectively followed low ratios did not reflect contact with the syndrome-associated retrovirus. Abnormal ratios were rarely seen in men who had never been exposed to cytomegalovirus. Thus, cytomegalovirus infection may be an important cofactor in the immunologic disorders leading to the acquired immunodeficiency syndrome. PMID- 2988393 TI - [Vitamin D deficiency in long-stay geriatric patients. Study of a therapeutic protocol adapted to the institutional milieu]. AB - A vitamin D deficiency was observed in 22 women and 6 men aged 66 to 94 years, hospitalised in a long-stay department in the Paris region for an average of 5 years and incapable of going out. The diagnosis was confirmed by measuring serum 25 (OH) D: 1.89 +/- 0.18 ng/ml during the month of June (11.5 +/- 4.2 ng/ml at the same period in II healthy young adults). The patients were randomly divided into two groups of 11 (Group I) and 17 patients (Group II). The first group was given an improved diet in calcium and Vit. D; the second group received, in addition, an intramuscular supplement of 30 mg of Vit. D3 in monthly injections of 5 or 10 mg. The serum 25 (OH) D levels were measured each month in both groups. Patients with intramuscular supplements showed a slow rise until the total injected dose exceeded 20 mg, followed by a sudden rise to values greater than 30 ng/ml. Eight months after the last injection, mean serum 25 (OH) D was still 24.7 +/- 4.6 ng/ml compared to only 3.6 +/- 3 ng/ml in Group I. Vitamin D deficiency in geriatric patients cannot be corrected by dietary supplement alone, but parenteral Vit. D3 using an easily applicable therapeutic protocol is effective and well tolerated at the suggested dosage. PMID- 2988394 TI - [Anatomo-clinical conference. Hopital de la Pitie-Salpetriere. Case n. 1 - 1985. Prolonged fever and lymphopenia in a 26-year-old woman]. PMID- 2988395 TI - Modeling the mechanism of peptide cleavage by thermolysin. PMID- 2988396 TI - [Angiographic and surgical control of the vertebral and carotid pedicles supplying giant paraganglioma of the skull base and neck]. AB - Results obtained after excision of large jugular paragangliomas are largely conditioned by the possibility of control of the different vascular pedicles, and existing possibilities are discussed in relation to 6 briefly presented cases. The internal carotid artery can either be occluded radiologically by balloon catheter and then embolized if the artery is surrounded by a tumor, or controlled surgically if the artery is in contact with the tumor only and provides only a small proportion of its vascular supply. Whenever internal carotid artery occlusion is envisaged a clamping test by balloon under local anaesthesia is carried out initially. The vertebral artery pedicles are interrupted during surgical approach to the artery above the atlas. The value of a 2 or even 3 stage operation for excision of very large tumors with posterior fossa extension is discussed. PMID- 2988397 TI - [Malignant chemodectoma of the intrapetrous facial nerve]. AB - A 30 year old man presented with a tumor of the intrapetrous portion of the facial nerve, found to be a malignant chemodectoma on investigation. Possible relations with tumors of the APUD system, as well as pathogenic and therapeutic aspects are discussed. PMID- 2988398 TI - [Synovial sarcoma with cervicopharyngeal expression]. AB - A case is reported of a patient with cervicopharyngeal synovial sarcoma, and the clinical characteristics and therapy of these very rare tumors discussed. It appears difficult to assess prognosis because of the very few cases documented. PMID- 2988399 TI - Adenocarcinoma of the extrahepatic biliary tree. AB - Increasing survival rates for carcinoma of the biliary tree could reflect the selection of patients for referral to a specialist centre as well as modern improvements in diagnosis and treatment. To determine the true incidence and outcome of biliary cancer, the records of 243 unselected Bristol patients were reviewed retrospectively over a 15-year period. Mean age was 64 years. Gallstones were associated in 38% of cases. Sixty-nine of 87 patients with gallbladder carcinoma were submitted to laparotomy. The operative (30-day) mortality rate was 56% and the one-year survival rate was 7%; one patient with an unexpected carcinoma is alive 7 years after cholecystectomy (1% five-year survival rate). Among 31 patients with carcinoma of the ampulla the operative mortality rate was 27%; one-year and five-year survival rates were 46% and 15%. Bile duct cancers (n = 125) arose proximal to the entry of the cystic duct (n = 63), distal to that point (n = 46) or diffusely (n = 16). For proximal cholangiocarcinoma the operative mortality rate was 38% and 1-year survival rate 17%; no patient lived for 2 years. For distal cholangiocarcinoma the operative mortality rate was 45% and one-year survival rate 26%; no patient lived for 3 years. Only palliative procedures were undertaken for diffuse cholangiocarcinoma; 2 patients survived 1 year (13%). Overall 5 patients with carcinoma of the extrahepatic biliary tree appear to have been cured (2%). PMID- 2988400 TI - High fibre diet in symptomatic diverticular disease of the colon. AB - Over the past decade fibre supplementation has achieved widespread acceptance in the management of symptomatic diverticular disease, although the efficacy of this treatment has been debated. We have conducted a retrospective review of 72 patients admitted to hospital with symptomatic diverticular disease over a ten year period in order to determine whether or not high fibre diet afforded protection against the development of complications, necessity for surgery or persistence of symptoms. Fifty-six patients were treated non-operatively, of these 43 received advice concerning a high fibre diet but only 31 patients complied. The 12 patients who failed to take additional fibre and the 13 patients who never received dietary advice (25 patients) formed the non high fibre group. Those treated with fibre supplementation fared significantly better in developing fewer complications and required less surgery (P less than 0.05). At the time of follow-up review patients on a high fibre diet reported significantly fewer symptoms (P less than 0.05). PMID- 2988401 TI - [Surgery of insulinoma under an artificial pancreas]. PMID- 2988402 TI - A continuous monitoring spectrophotometric method for the measurement of angiotensin-converting enzyme in human serum. AB - A continuous monitoring spectrophotometric method for the measurement of angiotensin-converting enzyme in human serum is described. It is based on the hydrolysis of furylacrylylphenylalanylglycylglycine and applied to an IL Multistat III centrifugal analyser. The method offers better precision and freedom from interference than a fluorimetric assay we have previously described. PMID- 2988403 TI - Central angiotensin converting enzyme blockade and salt preference in renovascular hypertensive rats. AB - Previous studies have shown that the spontaneously hypertensive rat (SHR) has a preference for 0.9% NaCl solution over water as a drinking fluid. This preference was decreased by chronic treatment of SHR with intracerebroventricular captopril (an angiotensin converting enzyme inhibitor). Although other strains of rats were compared to SHR, no studies, that we are aware of, have been reported in renal hypertensive rats. Wistar-Kyoto rats were sham operated or had a silver clip (i.d., 0.20 mm) placed on the left renal artery to produce renovascular hypertension. Three weeks later the rats were operated upon again to implant osmotic minipumps to deliver captopril or saline either into the right lateral brain ventricle (i.c.v.) or into the peritoneal cavity (i.p.). The rats had a choice of 0.9% NaCl or tap water during the study. Blood pressures were measured by a tail plysmographic method in the conscious rats. The rats that became hypertensive showed a marked preference for saline. Treatment with captopril i.p. (24 mumol/kg/day) stimulated preference for saline but i.c.v. treatment (24 mumol/kg/day) decreased the preference for saline despite reductions in blood pressure in both groups of renal hypertensive rats. These changes were not seen in renal hypertensive rats infused with saline. The results suggest that captopril's antihypertensive effect in this model of renal hypertension may be independent of the effects of the drug on preference to drink saline. PMID- 2988404 TI - Abnormalities in glutamatergic mechanisms in human Huntington's disease. AB - Of many glutamatergic parameters studied in human Huntington's disease, the following abnormalities were documented in the literature: Decreased glutamate content in cerebrospinal fluid and frontal cortex. Decreased activities of glutamine synthetase, ornithine aminotransferase and aspartate aminotransferase in various brain regions, especially the frontal cortex, caudate nucleus and putamen. Decreased glutamate binding in fibroblast membranes. Although it has been hypothesized that Huntington's disease may have a glutamate-related etiology, presently available evidence is too fragmented and inadequate for any conclusion to be made. However, it is noted that interpretation of these human data is very difficult because of two reasons. Firstly, changes observed in postmortem Huntington's disease brains may only be secondary to the disease instead of being the cause of the disease. Secondly, there is always doubts as to the relevance of data obtained with non-neural tissues such as fibroblasts and platelets. PMID- 2988405 TI - Auditory brainstem responses in normal hearing ears. AB - It is essential to know the expected auditory brainstem response latencies of a normal hearing population to utilise this procedure as a diagnostic tool. Absolute wave latencies of the auditory brainstem responses were recorded on 50 normal hearing subjects, aged 3 months to 56 years using a standard sound pressure level of 115 dB alternate clicks. The recordings were examined for differences in latencies between the age groups, sexes and the left and right ears. The results showed that the absolute latencies of subjects aged 2 years and above were the same and no significant differences were obtained between the left and right ears. The infants showed longer latencies. The brainstem transmission time values were calculated as from Wave NI-NIII, Wave NI-NV and Wave NIII-NV. A comparison of the brainstem transmission times between the left and right ears of each individual subject showed that in all cases the differences did not exceed more than 0.2 msec. A value of more than 0.2 msec between the two ears of an individual may be of diagnostic importance. In comparing the male and female brainstem transmission times, there was a significant difference statistically. Another value to use was the Wave PV, which is the most prominent and stable peak. The results showed that the Wave PV latency progressively shortens during infancy to attain the adult value somewhere between 18 and 24 months. PMID- 2988406 TI - Biochemistry of anxiety. AB - The identification of a biochemical (cellular, molecular) basis of anxiety has not only a heuristic value but also considerable consequences for any kind of prophylaxis and therapy of anxiety. An abnormally high activity of noradrenergic and/or serotoninergic neuronal pathways, all originating in the lower brainstem has been proposed by various authors to be critical for the activation of neuronal networks generating anxiety. Recent progress in the understanding of the mechanism of action of anxiolytic drugs and of a variety of anxiogenic agents suggests the possibility that an abnormal neuronal activity of the nonconvulsive type in the limbic system generates the emotion of anxiety and the accompanying behavioral, autonomic and motor symptoms. A central role is suggested for gamma aminobutyric acid (GABA), which normally prevents those types of neurones possessing the inherent properties for paroxysmal activity to respond with epileptiform activity to normal or excessive stimuli. The GABA receptor/chloride channel complex with its regulatory sites may be the site of a primary biochemical functional lesion. PMID- 2988407 TI - The role of the FtsZ protein (SfiB) in UV-induced division inhibition and in the normal Escherichia coli cell division cycle. AB - This paper describes some of the major characteristics of the SOS-dependent division arrest which occurs in Escherichia coli during repair of DNA damage following UV irradiation. We shall review the evidence that the inducible division inhibitor, SfiA, interacts directly with an essential division protein, FtsZ. On the basis of its pivotal role in division inhibition during the UV stress response and other properties of ftsZ, we propose that this is a key gene involved in the actual regulation of the division cycle in E. coli. We also propose that at least some components of the division machinery interact to form a specific complex which we designate as a "septalsome". We shall discuss the possibility that a critical concentration of FtsZ is required to trigger the formation of an active septalsome prior to division. Alternatively, FtsZ might act to inhibit the formation of an active septalsome until a critical point in the cell cycle is reached. PMID- 2988408 TI - [Presence of plasmids in Neisseria meningitidis]. AB - The plasmid content of twenty-nine Neisseria meningitidis strains, isolated from CSF and blood cultures, was investigated by agarose gel electrophoresis. The strains (twenty-seven from serogroup B and two from serogroup C) were sensitive to antibiotics, but 50% were resistant to sulphonamide (MIC greater than or equal to 16 mg/l). Two serogroup-B strains from CSF harboured an extrachromosomal element. These two cryptic plasmids differed in size: 8.4 Kb for pNMT1 and 82 Kb for pNMT2. PMID- 2988409 TI - Inflammatory myopathies: Part 2. PMID- 2988410 TI - Increased prevalence and titer of Epstein-Barr virus antibodies in patients with multiple sclerosis. AB - The prevalence and titer of serum antibodies to several Epstein-Barr virus (EBV) antigens were compared among patients with multiple sclerosis, healthy siblings of multiple sclerosis patients, patients with other neurological diseases, and healthy non-blood-related subjects. Serum-cerebrospinal fluid (serum-CSF) pairs were available on a selected number of multiple sclerosis and control subjects. An increased antibody response to EBV antigens was noted rather consistently in the sera of the multiple sclerosis group in comparison with the control groups. A greater number of reduced ratios of serum:CSF IgG antibody to EBV-capsid antigen and antibody to EBV-early antigen components than to adenovirus, a reference or control virus, were found in the multiple sclerosis group. Reduced ratios of these EBV antibodies were detected more frequently or showed a trend in this direction in multiple sclerosis patients compared with the group with other neurological diseases. Our findings extend the results of an earlier report and strengthen the association between EBV and multiple sclerosis. PMID- 2988411 TI - Reversible, late-onset disulfiram-induced neuropathy and encephalopathy. AB - Disulfiram toxicity is a well-recognized cause of peripheral neuropathy and encephalopathy, usually developing within a few months of the start of therapy. We describe a patient who insidiously developed a peripheral neuropathy and encephalopathy after thirty years of disulfiram ingestion. Both complications partially resolved after the medication was stopped. PMID- 2988412 TI - Fatal infantile mitochondrial myopathy and renal dysfunction caused by cytochrome c oxidase deficiency: immunological studies in a new patient. AB - A 3-month-old female infant had profound generalized weakness, de Toni-Fanconi Debre syndrome, and lactic acidosis. She required assisted ventilation and died at the age of 8 months. Muscle biopsy showed accumulation of mitochondria, glycogen, and lipid droplets. Histochemical reaction and immunocytochemical stain for cytochrome c oxidase showed very weak results, but both reactions were normal in intrafusal fibers of the muscle spindle. In crude extracts of the patient's muscle, cytochrome c oxidase activity was undetectable and enzyme-linked immunosorbent assay showed decreased reaction at all dilutions of antiserum. These data indicate that the amount of immunoreactive enzyme protein is markedly decreased in muscle of patients with fatal infantile cytochrome c oxidase deficiency and renal dysfunction. PMID- 2988413 TI - Experimental lead neuropathy: inorganic lead inhibits proliferation but not differentiation of Schwann cells. AB - Schwann cells were prepared from the sciatic nerves of newborn rats and cultured in a monolayer. Addition of lead acetate at concentrations between 0.4 and 10.0 micrograms/ml, levels comparable to those occurring in neural tissues and physiological fluids of lead-intoxicated rats, diminished both the baseline rate of proliferation of the Schwann cells and their response to the mitogens, axolemmal fragments, glial growth factor, and the adenosine 3':5'-cyclic monophosphate (cAMP) analogues 8-bromo-cAMP and dibutyryl-cAMP. This demonstrates a direct toxic effect of inorganic lead on Schwann cells. Lead acetate in this concentration range did not, however, inhibit the cAMP analogue-induced appearance of the "myelin marker" lipid galactocerebroside on the surfaces of the cultured Schwann cells. PMID- 2988414 TI - Progressive encephalopathy in children with acquired immune deficiency syndrome. AB - We are reporting the cases of four children with a diagnosis of acquired immune deficiency syndrome (AIDS) who demonstrated a progressive encephalopathy manifested by loss of motor milestones or intellectual abilities, and weakness with pyramidal tract signs. Two patients were ataxic. One child was cortically blind and suffered myoclonic jerks before his death. Two of the patients had isolated seizures at some time during the disease course. Two patients developed secondary microcephaly and all four patients showed cortical atrophy on computed tomographic scanning. Three of the patients died. Postmortem neuropathological findings included gross cortical atrophy, microglial nodules, and intranuclear inclusions, as are described in the subacute encephalitis reported in adult AIDS patients. We propose that the possible pathogenetic mechanisms of this encephalopathy include opportunistic or persistent viral infection of the brain. PMID- 2988415 TI - Neurotransmitter receptors in amyotrophic lateral sclerosis: possible relationship to sparing of eye movements. PMID- 2988416 TI - The pharmacology of eating behavior. PMID- 2988417 TI - Phosphatidylinositol turnover in receptor mechanism and signal transduction. PMID- 2988418 TI - The pharmacological and physiological role of cyclic GMP in vascular smooth muscle relaxation. PMID- 2988419 TI - Affinity labels for opioid receptors. PMID- 2988420 TI - Synthetic and fermentation-derived angiotensin-converting enzyme inhibitors. PMID- 2988421 TI - The mechanisms of antiinflammatory steroid action in allergic diseases. PMID- 2988422 TI - The physiology and pharmacology of spinal opiates. PMID- 2988423 TI - Steroid hormone regulation of gene expression. PMID- 2988424 TI - The function of myosin and myosin light chain kinase phosphorylation in smooth muscle. PMID- 2988425 TI - Biochemical and genetic studies of iron-regulated (exotoxin A) and phosphate regulated (hemolysin phospholipase C) virulence factors of Pseudomonas aeruginosa. PMID- 2988426 TI - Contribution of exoenzyme S to the virulence of Pseudomonas aeruginosa. AB - Exoenzyme S is an extracellular ADPR transferase produced by P. aeruginosa. Forms of this enzyme that have thus far been purified are not toxic, however, exoenzyme S clearly contributes to the virulence of strain 388. Thus, a Tn1 mutant deficient in exoenzyme S was found to be markedly less virulent than its exoenzyme S-producing parental strain in both a burned mouse infection model and a rat chronic lung infection model. Exoenzyme S does not appear to contribute to initial colonization of the rat lung or the burned mouse skin. Exoenzyme S does, however, appear to contribute to local tissue damage in the rat lung, and to dissemination of P. aeruginosa from the skin into the blood and distant organs of the burned mouse. Perhaps our most important observation is that specific antibody against exoenzyme S confers a high level of protection against subsequent infection of burned mice. While these results must be extended to include additional strains they are encouraging, and they underscore the relative importance of exoenzyme S in the pathogenesis of P. aeruginosa infections. PMID- 2988427 TI - The Pseudomonas aeruginosa outer membrane permeability barrier and how to overcome it. AB - The intrinsic resistance of P. aeruginosa to most hydrophilic antibiotics can be explained, in part, on the basis of its low outer membrane permeability. Protein F which constitutes the major outer membrane porin protein for the uptake of hydrophilic compounds, is poorly functional. We have demonstrated that less than 1% of the 200,000 or so copies of protein F per cell can form active functional channels. Our working hypothesis is that the species of LPS associated with individual protein F trimers determines whether these trimers adopt a functional conformation. Since low outer membrane permeability constitutes a major problem for the penetration of antibiotics into P. aeruginosa, we have started to search for compounds which permeabilize outer membranes ('permeabilizers') and thus would be potentially synergistic with antibiotics. Eighteen permeabilizer compounds have been discovered and fall into defined chemical groupings including polycations, organic cations and divalent cation chelators. PMID- 2988428 TI - Evidence for a disseminated erythromycin resistance determinant mediated by Tn917 like sequences among group D streptococci isolated from pigs, chickens, and humans. AB - A total of 199 streptococci isolated from feces of healthy chickens, pigs, and beef cattle and 26 human clinical isolates were tested for resistance to kanamycin, streptomycin, tetracycline, erythromycin, and lincomycin. Of 66 isolates resistant to these antibiotics, 12 transferred one or more resistance traits by conjugation in broth. Erythromycin resistance (Emr) was transferred from 10 of the 12 successful donors. AvaI digests of plasmids isolated from Emr transconjugants derived from two human, two chicken, and one pig isolate contained three fragments similar in size to those produced from Tn3871, an Emr transposon. The three fragments from each of the five digests on Southern blots hybridized to radiolabeled Tn3871. Plasmid DNA from a transconjugant derived from a second pig isolate contained two of the three Tn3871-associated AvaI fragments. One of the AvaI fragments from each of the six plasmids hybridized with a radiolabeled probe containing a cloned AvaI fragment from Tn3871 that contained the Emr determinant. Transposition of the Emr trait was demonstrated for the plasmids derived from one human and one pig isolate. We concluded that extensive DNA homology existed between plasmids from streptococcal strains obtained from two human patients, two chickens, and two pigs and the Emr transposon Tn3871, which is very similar or identical to the well-characterized Emr transposon Tn917. The detection of Tn3871-like sequences in streptococcal isolates from Arkansas, Illinois, North Carolina, and Washington, D.C. indicates wide dissemination of Emr mediated by the same or closely related transposons. PMID- 2988429 TI - Mutually exclusive inhibition of herpesvirus DNA polymerase by aphidicolin, phosphonoformate, and acyclic nucleoside triphosphates. AB - Dual inhibitor studies were performed to examine the interaction of aphidicolin, phosphonoformate, 9-(2-hydroxyethoxymethyl)guanine triphosphate, and 9-(1,3 dihydroxy-2-propoxymethyl)guanine triphosphate with herpes simplex virus DNA polymerase. Kinetic data indicated that inhibition by one agent prevents simultaneous inhibition by a second agent, producing a mutually exclusive inhibition pattern. This suggested that binding sites on the DNA polymerase molecule for these compounds are kinetically overlapping. These findings should be taken into consideration for the design of future antiviral compounds and combination chemotherapy protocols. PMID- 2988430 TI - Binding of polymyxin B nonapeptide to gram-negative bacteria. AB - The binding of the outer membrane-disorganizing peptide polymyxin B nonapeptide (PMBN) to gram-negative bacteria was studied by using tritium-labeled PMBN. Smooth Salmonella typhimurium had a binding capacity of ca. 6 nmol of PMBN per mg (dry weight) of bacteria, which corresponds to ca. 1 X 10(6) to 2 X 10(6) molecules of PMBN per single cell. The binding was of relatively high affinity (Kd, 1.3 microM). The isolated outer membrane of S. typhimurium bound ca. 100 nmol of PMBN per mg of outer membrane protein (Kd, 1.1 microM), whereas the cytoplasmic membrane bound 9 to 10 times less. Other bacteria which are susceptible to the action of PMBN (Escherichia coli strains, Pseudomonas aeruginosa, Haemophilus influenzae) also bound large amounts of PMBN. The S. typhimurium pmrA mutant, Neisseria gonorrhoeae, and Proteus mirabilis (all known as resistant to polymyxin and PMBN) bound 3.3, 4, and 12 times less than S. typhimurium, respectively. The binding of PMBN to S. typhimurium was effectively inhibited by low concentrations of polymyxin B, compound EM49 (octapeptin), polylysine, and protamine. Spermine, Ca2+, and Mg2+ also inhibited the PMBN binding although they were ca. 160, 700, and 2,400 times less active (based on molarity) than polymyxin B, respectively. No binding inhibition was found at the tested concentrations of streptomycin, tetralysine, spermidine, or cadaverine. PMID- 2988431 TI - Comparative studies on the modes of action of the antirhinovirus agents Ro 09 0410, Ro 09-0179, RMI-15,731, 4',6-dichloroflavan, and enviroxime. AB - Modes of action of five antirhinovirus agents were compared. Ro 09-0410, 4',6 dichloroflavan, and RMI-15,731 were active preferentially against human rhinovirus. Serotypes of the virus varied in their susceptibility to these three agents, whereas Ro 09-0179 and enviroxime showed activity against all the serotypes of the virus tested to date. Ro 09-0410, RMI-15,731, and 4',6 dichloroflavan inactivated the virus directly, although 4',6-dichloroflavan did so only slightly. Inactivation by 4',6-dichloroflavan and RMI-15,731 was associated with the binding of the agents to the virus, since the infectivity, reduced by exposure to the agents, was restored to the original level by extraction of the agents with chloroform. The binding of [3H]Ro 09-0410 to human rhinovirus type 2 was inhibited by unlabeled Ro 09-0410, 4',6-dichloroflavan, and RMI-15,731 but not by Ro 09-0179 or enviroxime. Furthermore, subtypes resistant to both 4',6-dichloroflavan and RMI-15,731 showed cross-resistance to Ro 09-0410 and vice versa. On the other hand, sublines resistant to these three agents were not cross-resistant to Ro 09-0179 or enviroxime. These results indicate (i) that Ro 09-0410, 4',6-dichloroflavan, and RMI-15,731 exert their activities through the same mode of action, namely, binding to or interaction with some specific site on the viral capsid protein, and (ii) that the binding or interaction sites for these three agents are either the same or very close to each other. PMID- 2988432 TI - Inhibition of human cytomegalovirus by combined acyclovir and vidarabine. AB - The inhibition of human cytomegalovirus (HCMV) isolates by acyclovir (ACV) and vidarabine (ara-A) was assessed by using an infectious-center plaque-reduction assay. When fixed concentrations of 4.5 micrograms of ACV and 250 ng of ara-A per ml were compared singly and in combination, the viral inhibition resulting from the ACV-ara-A combination was synergistic for three of four HCMV clinical isolates studied and additive for one HCMV isolate. An additional four HCMV strains obtained at postmortem examination from the lungs of bone marrow transplant patients were assessed for sensitivity to ACV-ara-A by using the dose required for 50% viral inhibition (ID50) as the endpoint. The mean ID50 of ACV for the four HCMV isolates was 12.3 micrograms/ml, whereas the mean ID50 of ara-A was 3.4 micrograms/ml. When 1 microgram of ara-A per ml (which yielded a mean plaque reduction of 23.6%) was combined with ACV, a mean of 5.2 micrograms of ACV per ml was required for 50% viral inhibition. The sum of the fractional inhibitory concentrations for each of the four HCMV isolates was less than 1, indicating synergy by the ACV-ara-A combination. Although DNA synthesis in growing human embryonic lung fibroblast (HEL) cells, as determined by [3H]thymidine incorporation, was diminished to 61% of that in untreated control cells when 22.5 micrograms of ACV and 1 microgram of ara-A per ml were used, there was no additive inhibition of DNA synthesis when the two-drug combination was used. HEL cell growth remained at 97% of control cell growth at 72 h when concentrations as high as 45 micrograms of ACV combined with 1 microgram of ara-A per ml were used. PMID- 2988433 TI - Inhibitory effect of cephalosporins on gamma-aminobutyric acid receptor binding in rat synaptic membranes. AB - Cephalosporins inhibited gamma-aminobutyric acid receptor binding in a concentration-dependent manner in vitro. Scatchard analysis revealed that cefazolin decreased the binding capacity but did not change the affinity of the receptor. It is suggested that this inhibition of gamma-aminobutyric acid receptor binding may be involved in the induction of convulsions by cephalosporins. PMID- 2988434 TI - Correlation of aminoglycoside resistance with the KmS and Vmax/Km ratios of enzymatic modification of aminoglycosides by 2''-O-nucleotidyltransferase. AB - A clinical isolate, Serratia marcescens 75, was found to be susceptible to netilmicin and yet had a high level of aminoglycoside 2''-O nucleotidyltransferase activity for netilmicin. Kinetic studies of the partially purified enzyme revealed substrate inhibition for gentamicin and tobramycin at concentrations greater than 10(-2) mM, but this was not observed for netilmicin. The MICs of the aminoglycosides tested exhibited a good inverse correlation with the Km values for the enzyme and a direct correlation with the Vmax/Km ratios of the enzyme. PMID- 2988435 TI - Influence of adsorption time, rocking, and soluble proteins on the plaque assay of monodispersed poliovirus. AB - Factors that could affect adsorption of monodispersed poliovirus to cell culture monolayers were evaluated. These included varying the virus adsorption period under static and nonstatic (rocked) conditions and altering the rocking rate. The effects of several soluble proteins on plaque formation, enumeration, and size were also evaluated. Rocking involved the mechanical spread of viruses over cell culture monolayers for 1 to 4 h. Rocked cultures exhibited significantly higher (P less than 0.05) plaque counts than corresponding static cultures. Optimal plaque counts were obtained after a 2-h adsorption period with rocking; increasing the period to 4 h did not significantly increase PFU. Optimal counts were not obtained until greater than or equal to 4 h with static adsorption. Plaque counts were not affected by increasing the rocking rate above one oscillation per minute, but a slower rocking rate resulted in a significant decrease in plaques. Adsorption of poliovirus in the presence of 3% solutions of beef and meat extracts, acid-precipitated oyster protein, two brands of skim milk, and 3 and 10% fetal bovine serum was compared with adsorption in protein free controls. Significant reductions (P less than 0.05) in plaque counts occurred with one brand of skim milk, whereas 3% beef extract yielded highly significant reductions (P less than 0.01) in plaque counts and appreciable decreases in plaque sizes. Salinities of protein-containing virus inocula were high for beef and meat extracts but somewhat below physiological levels for the remaining inocula. Beef extract-associated reductions in PFU were eliminated after the extracts were dialyzed. Plaque reductions were associated with dialyzable components of the beef extract but not with the inoculum salinity. PMID- 2988436 TI - Persistently infected cultures as a source of hepatitis A virus. AB - Primary African green monkey kidney, continuous African green monkey kidney cell line BS-C-1, and buffalo green monkey kidney cultures were infected with a uniform inoculum of hepatitis A virus (HAV). Although both the cell line BS-C-1 and primary African green monkey kidney cultures produced useful amounts of virus, HAV was detected earlier and in greater quantities in primary African green monkey kidney cultures. A persistently infected primary African green monkey kidney culture was developed. The influence of incubation time (4 to 40 days) and concentration (2 to 15%) of fetal calf serum in the maintenance medium on production of HAV by this culture was examined. An incubation period of 24 to 28 days was found to be optimal; reducing this period led to decreased yields of HAV. No significant difference in the amount of HAV produced was observed with differing concentrations of fetal calf serum. Three different methods of extraction and the effect of multiple extractions on the recovery of HAV from cell lysates were examined. Sonication was a critical factor. Two extractions yielded more than 90% recoverable virus. Yields in excess of 10(11) physical particles of HAV per 850-cm2 roller bottle were routine. The total yield could be increased by concentrating the HAV present in spent maintenance medium by using bentonite or organic flocculation. PMID- 2988437 TI - Plasmid-borne Tn5 insertion mutation resulting in accumulation of gentisate from salicylate. AB - Plasmid-borne Tn5 insertion mutants of a Pseudomonas species which accumulated 2,5-dihydroxybenzoate (gentisate) following growth on 2-hydroxybenzoate (salicylate) were obtained from a pool of mutants that were unable to grow on naphthalene. One such mutant was characterized further. The ability of this mutant to oxidize gentisate was 100-fold less than the ability of a Nah+ Sal+ strain harboring the unmutagenized plasmid, although both strains oxidized and grew on salicylate. These bacteria were presumably able to metabolize salicylate via catechol, since they possessed an inducible, plasmid-encoded catechol 2,3 dioxygenase. Our results suggest that there is an alternate, plasmid-encoded route of salicylate degradation via gentisate and that some plasmid-associated relationship between this pathway and naphthalene oxidation exists. PMID- 2988438 TI - Effects of bentonite clay solids on poliovirus concentration from water by microporous filter methods. AB - To determine whether suspended solids interfere with enteric virus recovery from water by microporous filter methods, the effects of bentonite clay solids at a concentration of 10 nephelometric turbidity units on the recovery of poliovirus type 1 from seeded, activated carbon-treated, filtered tap water were studied. Volumes (500 ml) of virus-laden water at pH 5.5 or 7.5, with and without 50 mM MgCl2, were filtered through 47-mm-diameter, electropositive (Virosorb 1MDS) and electronegative (Filterite) filters that had been pretreated with Tween 80 to minimize direct virus adsorption to filter surfaces. Bentonite solids enhanced virus retention on both types of filters, even under conditions in which viruses were not solids associated. However, bentonite solids also interfered with elution of retained viruses when eluting with 0.3% beef extract-50 mM glycine (pH 9.5). Under some conditions, overall virus recoveries were lower from water with bentonite solids than from solids-free control water. The results of this study indicate that clay turbidity can interfere somewhat with virus recovery by current microporous filter methods. PMID- 2988439 TI - Enumeration and activity of cellulolytic bacteria from gestating swine fed various levels of dietary fiber. AB - Cellulolytic bacteria were enumerated and cellulase activity was determined over a 98-day period from fecal samples of gestating swine fed various levels and sources of fiber. The diets, each fed to five pigs, were a corn-soybean control, 20% corn cobs, and 40 and 96% alfalfa meal. Fecal samples were collected from all pigs on days 0, 5, 14, 21, 35, 49, 70, and 98. Overall, the most probable number of cellulolytic bacteria from pigs fed the control, 20% corn cobs, and 40 and 96% alfalfa meal was 23.3 X 10(8), 15.2 X 10(8), 45.1 X 10(8), and 52.5 X 10(8) per g (dry weight) of fecal sample, respectively. Enumeration of cellulolytic bacteria by counting zones of clearing in roll tubes, as compared with the most probable number procedure, accounted for only 1.1 and 17.0% of the cellulolytic bacteria, respectively, from pigs fed the control or 96% alfalfa meal diet. Cellulolytic bacteria (most probable number) on days 70 and 98 accounted for 4.1 and 10.0% of the viable count for the pigs fed the control and 96% alfalfa meal diets, respectively. The viable count was not different between pigs fed the control and 96% alfalfa meal diets. The overall mean cellulolytic activity (milligrams of glucose released from carboxymethyl cellulose per gram [dry weight] fecal sample per 30 min was 17.0, 19.9, 23.8, and 20.6, respectively, for the control, 20% corn cobs, and 40 and 96% alfalfa meal diets. The results indicate that the cellulolytic flora can be increased by prolonged feeding of high-fiber diets and may represent 10% of the culturable flora. PMID- 2988441 TI - Relative resistance to chlorine of poliovirus and coxsackievirus isolates from environmental sources and drinking water. AB - Several poliovirus and coxsackievirus isolates from environmental sources were compared with laboratory strains to determine their rate of inactivation by chlorine. All viruses were tested for up to 1,000 min in the presence of an initial free residual chlorine level of ca. 0.4 mg/liter. Coxsackievirus B5 (CB 5) isolates were found to be more resistant to chlorine than coxsackievirus B4 (CB-4), followed by poliovirus 1, 2, and 3 in order of decreasing resistance to chlorine. Environmental isolates of CB-5 were more resistant than the laboratory strain tested, and for two strains 12 and 22% of the input virus was still infectious after 100 min in the presence of free residual chlorine. Although CB-4 isolates were less resistant to chlorine than CB-5 isolates, after 1,000 min of contact 0.01% of the input virus was still infectious. Except for CB-5 isolates, isolates from environmental sources did not appear to be more resistant to chlorine than laboratory strains. Viruses isolated at different phases during the preparation of drinking water were not more resistant to chlorine and must thus have been protected by other mechanisms. PMID- 2988440 TI - Metabolism of aflatoxin B1 by rat hepatic microsomes induced by polyhalogenated biphenyl congeners. AB - The metabolism of aflatoxin B1 to aflatoxins M1 and Q1 by rat liver microsomes from animals pretreated with polychlorinated or polybrominated biphenyl congeners depended on the structure of the halogenated biphenyl inducers. Microsomes from rats treated with phenobarbital (PB) or halogenated biphenyls that exhibit PB type activity preferentially enhanced the conversion of aflatoxin B1 to aflatoxin Q1. In contrast, microsomes from rats treated with 3-methylcholanthrene (MC) or halogenated biphenyls that exhibit MC-type induction activity increased the metabolism of aflatoxin B1 to aflatoxin M1. The coadministration of PB and MC produced microsomes that exhibited both types of induction activity (mixed type) in catalyzing the oxidative metabolism of diverse xenobiotic agents. However, PB plus-MC-induced hepatic microsomes from immature male Wistar rats preferentially increased the metabolism of aflatoxin B1 to aflatoxin M1 but did not enhance the conversion of aflatoxin B1 to aflatoxin Q1. Comparable results were observed with microsomes from rats pretreated with halogenated biphenyls classified as mixed type inducers; moreover, in some cases there was a significant decrease in the conversion of aflatoxin B1 to aflatoxin Q1 (compared with that of controls treated with corn oil). PMID- 2988442 TI - Effect of an activated sludge wastewater treatment plant on ambient air densities of aerosols containing bacteria and viruses. AB - Bacteria- and virus-containing aerosols were studied during the late summer and fall seasons in a midwestern suburb of the United States before and during the start-up and operation of an unenclosed activated sludge wastewater treatment plant. The study showed that the air in this suburban area contained low-level densities of indicator microorganisms. After the plant began operating, the densities of total aerobic bacteria-containing particles, standard plate count bacteria, total coliforms, fecal coliforms, fecal streptococci, and coliphages increased significantly in the air within the perimeter of the plant. Before plant operations, bacteria were detected from five genera, Klebsiella, Enterobacter, Serratia, Salmonella, and Aeromonas. During plant operations, the number of genera identified increased to 11. In addition to those genera found before plant operations, Escherichia, Providencia, Citrobacter, Acinetobacter, Pasteurella, and Proteus, were also identified. Enteric viruses were detected in low densities from the air emissions of this plant. Only standard plate count bacteria remained at significantly higher than base-line densities beyond 250 m downwind from the center of the aeration tanks. Fecal streptococci and coliphages appeared to be more stable in aerosols than the other indicator microorganisms studied. In general, the densities of microorganism-containing aerosols were higher at night than during the day. The techniques used in this study may be employed to establish microorganism-containing aerosol exposure during epidemiological investigations. PMID- 2988443 TI - Effect of sulfhydryl-disulfide state on protein phosphorylation: phosphorylation of bovine serum albumin. AB - Bovine serum albumin (BSA) was phosphorylated by the catalytic subunit of cAMP dependent protein kinase under general protein phosphorylation conditions. The optimal pH for this phosphorylation was 9.0. The K0.5 (the concentration required for 50% of maximal phosphorylation) for BSA at pH 7.5 was 15 microM. One mole of phosphate was incorporated per mole of BSA, and only one phosphopeptide fragment was obtained after extensive proteolysis with trypsin. BSA phosphorylation required dithiothreitol or GSH, but GSH was only one-fiftieth as effective as dithiothreitol. GSSG counteracted the effect of dithiothreitol and GSH. Phosphorylation increased in a time-dependent and dithiothreitol concentration dependent manner when BSA was preincubated with dithiothreitol. The increase in the incorporation of 32P correlated with the appearance of up to six free sulfhydryl groups. The effect of dithiothreitol on BSA appeared reversible, since reoxidation of reduced BSA decreased its susceptibility to phosphorylation. These experiments showed that this in vitro phosphorylation is dependent on the sulfhydryl-disulfide state of BSA. The possible implications of the sulfhydryl disulfide state of proteins in the regulation of phosphorylation are discussed. PMID- 2988444 TI - Nitroxides as redox probes of melanins: dark-induced and photoinduced changes in redox equilibria. AB - The interaction of nitroxide free radicals and their reduced products (hydroxylamines) with synthetic and natural melanins has been studied. Electron spin resonance spectroscopy was used to measure changes in radical concentration in the dark and during irradiation with visible or uv light. Some reduction of nitroxide occurs in the dark, and is reversible: the nitroxide can be completely regenerated by the one-electron oxidant ferricyanide. The kinetics of the process depend strongly on radical charge and pH. For positively charged nitroxides the rate is much faster than for either neutral or anionic radicals. At pH 10 the rate is about 20 times faster than at pH 5. Oxidation of hydroxylamine also can occur so that a redox equilibrium is established. The equilibrium constant has been estimated for the reaction between a nitroxide and melanin from autoxidation of 3,4-dihydroxyphenylalanine. Results are also dependent upon the type of melanin used and chemical modification (oxidation or reduction) of the melanin. Redox equilibria are altered during irradiation with either visible or uv light. Rapid oxidation of hydroxylamine to nitroxide is apparent, together with a slower reduction of nitroxide. Action spectra for these processes are related to those for melanin radical production and oxygen consumption in nitroxide-free melanin systems. Reduction of nitroxide is inhibited by oxygen, suggesting a competition between nitroxide and oxygen for photoinduced reducing equivalents. PMID- 2988445 TI - Immunoelectrophoretic characterization of the ADP/ATP carrier from heart, kidney, and liver. AB - Earlier studies gave an indication for an organ specificity of the ADP/ATP carrier. We used a modified charge-shift crossed immunoelectrophoresis for a more precise immunochemical characterization of this detergent-solubilized hydrophobic membrane protein. Immunological differences between the carrier protein from heart, kidney, and liver were demonstrated by a different electrophoretic migration of the three ligand-protein complexes in the first dimension and a distinct staining intensity, sharpness, and shape of the precipitates in the second dimension. However, the antibodies against the heart and kidney protein showed a cross-reactivity between the three antigens. The results are consistent with the view that the ADP/ATP carrier has organ-specific antigenic determinants although there is a partial identity between the carrier proteins from heart, kidney, and liver. PMID- 2988446 TI - Chlorination of NADH: similarities of the HOCl-supported and chloroperoxidase catalyzed reactions. AB - The chloroperoxidase-catalyzed reactions of NAD(P)H with H2O2 in the presence of Cl- or Br- have been characterized. With 1 mol H2O2 per mol of NADH, one atom of 36Cl was incorporated into the 264-nm-absorbing intermediate product. This species was oxidized enzymatically by a second mole of H2O2 to a species distinct from NAD+, which retained one Cl atom. Spectroscopically identical species were also produced by reaction of NADH with one and two molar ratios of HOCl, respectively. These data indicate that, with respect to halogenation activities, chloroperoxidase functions similarly to myeloperoxidase, i.e., produces HOCl as the first product of Cl- oxidation by H2O2. Moreover, rapid chlorination of NAD(P)H followed by oxidation may be an important and highly lethal microbicidal effect of HOCl produced by myeloperoxidase in activated neutrophils. PMID- 2988447 TI - Evidence for an aldehyde intermediate in the catalytic mechanism of thiamine oxidase. AB - Thiamine oxidase catalyzes the four-electron oxidation of the 5-hydroxyethyl group of thiamine to form thiamine acetic acid via an aldehyde intermediate. Evidence for the formation of this intermediate is derived from a number of kinetic approaches. The rate of thiamine acetic acid formation, as monitored by the rate of proton release, is subject to substrate inhibition and to inhibition by the presence of semicarbazide while the rate of O2 consumption (due to thiamine oxidation to the aldehyde and subsequently to the carboxylic acid) is unaffected. The transient formation of an intermediate with a maximal absorption at 370 nm in stopped-flow turnover experiments is dependent on the pH and the substrate concentration, and is prevented by the presence of semicarbazide, thus suggesting this transient absorption intermediate to be a result of formation of the aldehyde intermediate. A similar spectral intermediate is observed when hydroxythiamine is the substrate but is not observed with pyrithiamine. In the presence of large concentrations of pyrithiamine, the enzyme undergoes an irreversible inactivation which is not reversed on removal of pyrithiamine or its oxidation products by gel filtration or dialysis. This inhibition is prevented by the presence of thiols or of semicarbazide and is suggested to be due to the release of the aldehyde form of pyrithiamine from the catalytic site, which then reacts with the enzyme in a nonspecific manner. The structure of the 370-nm absorbing intermediate is currently unknown but is suggested not to be the "yellow form" of thiamine. This suggestion is due to observed differences in absorption spectral properties and to the fact that it can also be formed from hydroxythiamine, which does not form the "yellow form" of thiamine on alkaline treatment. Taken together, these data suggest that, at or below saturating concentrations, thiamine remains bound to the catalytic site during the two sequential two-electron transfer steps, with 2 mol O2 being reduced to 2 mol H2O2. At high concentrations (greater than 10 Km), the intermediate thiamine aldehyde can be displaced from the catalytic site by thiamine simply by a mass action effect. PMID- 2988448 TI - Properties and reaction mechanism of C4 leaf pyruvate,Pi dikinase. AB - The properties and reaction mechanism of maize leaf pyruvate,Pi dikinase are described. Km values were determined for the forward reaction substrates, pyruvate, ATP, and Pi, at pH 7.4 and 8.0 and for reverse reaction substrates at pH 7.4. Enzyme activity was almost totally dependent on added monovalent cations in both directions. NH+4 was most effective, with Ka values of about 0.38 mM for the forward reaction and 2 mM for the reverse reaction. K+ also completely activated the enzyme in the forward direction (Ka = 8 mM) but only partially activated in the reverse direction. Na+ had little effect on either reaction. The pH optimum for the forward reaction was about 8.2; the reverse reaction optimum was about 6.9. Maximum activity for the reverse direction was about twice the maximum forward direction rate. From data on the requirements for the ATP-AMP exchange reaction, on the mechanism of inhibition of the forward reaction by PEP, AMP, and PPi, and from the kinetics of the interaction of varying certain substrate pairs, it was concluded that the maize leaf pyruvate,Pi dikinase reaction proceeded by the two-step Bi Bi Uni Uni mechanism. This differs from the mechanism of catalysis by the bacterial enzyme. PMID- 2988450 TI - Chloroplast DNA synthesis during the cell cycle in cultured cells of Nicotiana tabacum: inhibition by nalidixic acid and hydroxyurea. AB - The effects of nalidixic acid and hydroxyurea on nuclear and chloroplast DNA formation in cultured cells of Nicotiana tabacum were investigated. At low concentrations (5 and 20 micrograms/ml) nalidixic acid, an inhibitor of DNA gyrase, exhibited a greater inhibitory effect on plastid DNA synthesis than on nuclear DNA formation. Since the plastid genome is a circular double-stranded DNA, this is consistent with the proven involvement of a DNA gyrase in the replication of closed circular duplex DNA genomes in procaryotic cells. At a high concentration of nalidixic acid (50 micrograms/ml), DNA synthesis in both the plastid and nuclear compartment was rapidly inhibited. Removal of the drug from the culture medium led to the resumption of DNA synthesis in 8 h. Hydroxyurea, an inhibitor of ribonucleoside diphosphate reductase, also depresses nuclear as well as plastid DNA formation. Removal of hydroxyurea from the blocked cells leads to a burst of nuclear DNA synthesis, suggesting that the cells had been synchronized at the G1/S boundary. The recovery of plastid DNA synthesis occurs within the same time frame as that of nuclear DNA. However, whereas plastid DNA formation is then maintained at a constant rate, nuclear DNA synthesis reaches a peak and subsequently declines. These results indicate that the synthesis of plastid DNA is independent of the cell cycle events governing nuclear DNA formation in cultured plant cells. PMID- 2988449 TI - Cloning and expression of the metE gene in Escherichia coli. AB - A lambda-transducing phage was isolated that contains the metE gene. This gene codes for N5-methyl-H4-folate:homocysteine methyltransferase (EC 2.1.1.14), an enzyme that catalyzes the terminal reaction in methionine biosynthesis. A 9.1-kb EcoR1 fragment of this phage, containing the metE gene, was then cloned into pBR325. This plasmid, pJ19, was used to transform Escherichia coli strain 2276, a metE mutant, and restore the MetE+ phenotype. Although the transformed cells produced large amounts of the metE protein in vivo, in vitro studies using pJ19 as template showed low synthesis of the metE protein. PMID- 2988451 TI - Structural changes of rabbit liver fructose-1,6-bisphosphatase: the effect of urea and subtilisin digestion. AB - At pH 6.3 both the native and subtilisin-digested fructose-1,6-bisphosphatase (Fru-P2-ase) molecules exhibit four fast-reacting thiol groups. The kinetic analysis shows that the pK value for the reaction of these thiols is 8.1. The increase of pH from 6.3 to 9.3 results in an uncovering of the remaining 20 thiol groups. In subtilisin-cleaved enzyme the rate of reaction of SH groups is considerably higher than in the native enzyme at pH 9.3, indicating changes in the microenvironments around thiols upon modification. A fluorescent label inserted on a fast-reacting SH group and neighboring NH2 group shifts the pH optimum of the enzyme to alkaline region and decreases its sensitivity toward AMP. Spectral analysis of labeled enzyme indicates that the labeled region of protein is more hydrophilic upon proteolytic digestion. It is concluded that a molecule of subtilisin-digested enzyme has a more relaxed structure than the native enzyme. The relaxation of the enzyme to a new conformation is reflected by urea addition, which mimics the effect of subtilisin digestion. Correlation of enzyme activity versus its sensitivity toward AMP (I 0.5), shows that at low concentrations of urea the active-site region at pH 6.3 is more affected than the region of AMP binding. PMID- 2988452 TI - Aldose reductase inhibitors: flavonoids, alkaloids, acetophenones, benzophenones, and spirohydantoins of chroman. AB - The inhibitory activity of various compounds, including 12 flavonoids, 10 alkaloids, 15 benzophenones, 5 acetophenones, and 7 spirohydantoins of chroman, was tested on rabbit lens aldose reductase, an enzyme involved in complications of diabetes. Almost all compounds tested were found to inhibit the enzyme at low concentrations (10(-5) M). The most potent inhibitor was 2R,4S-6-chloro-2 methylspiro(chroman-4,4'-imidazo-lidine+ ++)-2',5'-dione with an I50 value of 4.7 x 10(-8) M; other spirohydantoins showed similar potency. Polyhydroxybenzophenones were also potent inhibitors with an I50 value of about 10(-7) M. The possible structure-inhibitory activity relationships of the compounds tested are discussed. PMID- 2988453 TI - The Golgi apparatus and adrenal function: the effects of monensin on adrenocorticotropic hormone-stimulated steroidogenesis. AB - It has previously been shown that the steroidogenic action of adrenocorticotropic hormone (ACTH) is accompanied by characteristic alterations in cell ultrastructure. These include hypertrophy of the Golgi complex associated with increased vesicle formation and striking elevations of acid phosphatase activity in the Golgi complex and lysosomes. To investigate a possible relationship of these phenomena to steroidogenic function in monolayer cultures of murine adrenal tumor cells, monensin, a carboxylic ionophore which disrupts the ordered structure and transport function of the Golgi complex, was used. Monensin, at a concentration of 1.2 microM, causes massive vacuolization and hypertrophy of the Golgi complex. No effect on mitochondrial structure was seen. Monensin, 0.6-1.2 microM, inhibits both ACTH-stimulated and basal steroidogenesis by approximately 50% in incubations of 4 h or less. Dibutyryl-cAMP-stimulated steroidogenesis was inhibited to a similar degree. Incubations were carried out in serum-free media to eliminate possible effects due to exogenous cholesterol transport into the cell. There were no direct inhibitory effects of monensin on cholesterol side chain cleavage (SCC) activity in isolated mitochondria. In contrast, mitochondria isolated from cells previously treated with monensin had a reduced capacity for this activity. These experiments suggest that monensin inhibits transport of cholesterol from the Golgi complex to the mitochondrial site of steroidogenesis action or interferes with the transport of key mitochondrial proteins synthesized on cytoplasmic ribosomes. PMID- 2988455 TI - [Phase II study of cis-dichlorodiammineplatinum (II) for non-small cell lung cancer]. AB - A phase II study of cis-dichlorodiammineplatinum (II) (CDDP) was performed in 83 patients with inoperable non-small cell lung cancer (21 squamous cell carcinomas, 57 adenocarcinomas and 5 large cell carcinomas). CDDP was given by i.v. infusion at a dosage of 50 mg/m2 (low-dose regimen) or 80-100 mg/m2 (high-dose regimen) every 3 to 4 weeks. Seventy-nine out of 83 patients were evaluable for tumor response. Sixteen of these evaluable patients achieved complete or partial response, and the overall response rate was 20.3%. The response rates were 27.8% in patients with squamous cell carcinoma and 19.6% in those with adenocarcinoma. None of the patients with large cell carcinoma responded. The high-dose regimen was superior to the low-dose regimen in response rate (25.6% versus 5.8%). The responders survived significantly longer than patients with NC (p = 0.02) or PD (p = 0.003). Leukopenia of less than 3000/mm3 occurred in 12.1% of cases, and thrombocytopenia of less than 7 X 10(4)/mm3 occurred in 7.6%. A transient elevation of serum creatinine value was observed in 9 patients (10.8%). Moderate to severe nausea and/or vomiting occurred in almost all patients. It was therefore considered that CDDP was one of the most effective agents for non-small cell lung cancer with tolerable toxic effects. PMID- 2988454 TI - The reversible activation by Mn2+ ions of the Ca2+-requiring neutral proteinase of human erythrocytes. AB - Mn2+ (50 microM) satisfies the requirement for activity of the purified Ca2+ dependent neutral proteinase from human erythrocytes. Unlike the activation by Ca2+ [E. Melloni et al. (1984) Biochem. Int. 8, 477-489], the effect of Mn2+ is fully reversible and does not involve autodigestion of the native 80-kDa catalytic subunit. However, the native dimeric proenzyme (procalpain), which contains both the 80-kDa subunit and a smaller 30-kDa subunit, is not activated by Mn2+ alone but also requires the presence of micromolar concentrations of Ca2+. Under these conditions, 40% of the maximum activity is expressed without dissociation of the 80- and 30-kDa subunits. Mn2+, but not micromolar Ca2+, can also partially satisfy the metal requirement of the native 80-kDa subunit isolated after dissociation of the heterodimer. This activity is further enhanced by the addition of 5 microM Ca2+, which is ineffective in the absence of Mn2+. After procalpain is converted to active calpain by incubation with Ca2+ and substrate [S. Pontremoli et al. (1984) Biochem. Biophys. Res. Commun. 123, 331 337] full activity is observed with 5 microM Mn2+, which now substitutes completely for Ca2+. Activation of procalpain by Mn2+ represents a new mechanism for modulation of the Ca2+-dependent proteinase activity. PMID- 2988456 TI - [Antitumor effect of human interferon-alpha A/D in mice (II). Activation of natural killer cells and suppression of metastasis]. AB - Human recombinant interferon-alpha A/D (IFN-alpha A/D) is known to be effective on murine cells. We studied the in vivo effects of pure IFN-alpha A/D on murine splenic natural killer (NK) activity and experimentally induced pulmonary metastasis of B16-F10 melanoma. Intraperitoneal injection of IFN-alpha A/D augmented splenic NK activity, and suppressed melanoma metastasis. This suppression was abrogated by pretreatment of mice with anti-asialo GMl, but not with silica or carrageenan, showing that the effect of IFN-alpha A/D was dependent on NK cells. The suppression was strongest when IFN-alpha A/D treatment was given 12 hrs before or at the same time as tumor injection, showing that NK cells are most effective in the early phase of melanoma metastasis. PMID- 2988458 TI - [Analysis of human sera obtained from lung cancer patients by two-dimensional electrophoresis after schizophyllan (SPG) treatment]. AB - Schizophyllan (SPG) was administered to 13 lung cancer patients (i.m. 20mg X 2/week) for 3 weeks without chemo or irradiation therapies, and serum proteins were analyzed by two-dimensional electrophoresis (TDE). Additionally, immunosuppressive acidic protein (IAP) was quantitatively determined by single radial immunodiffusion (SRID). By TDE analysis, human serum proteins were separated into more than 100 spots, and about 14 spots were found to show quantitative changes in cancer patients. Quantitative examination was therefore conducted on changes of 8 components among these spots, including alpha 1-acidic glycoprotein (alpha 1 AG), acidic alpha 2-macroglobulin (acidic alpha 2 M), haptoglobin (Hp) and IAP. The protein which showed the most marked decrease in cancer patients, located between transferrin and IgG on the above TDE patterns, was ascertained to have a molecular weight of about 150,000 using a gel filtration method. This protein was increased in 7 of 13 patients after SPG treatment. PMID- 2988457 TI - [Phase III controlled studies of bestatin in malignant tumors of the skin- results of treatment of squamous cell carcinoma and genital Paget's disease]. AB - We have performed a co-operative controlled clinical study of Bestatin in the treatment of malignant tumors of the skin at 18 research institutions in Japan. Twenty-seven Bestatin-treated cases of stage I and II squamous cell carcinoma and 24 control cases were followed up for a maximum of 46 months, to compare disease free and survival rates in the 2 groups. There was no significant difference in either rate between the groups, though each rate was slightly higher for the Bestatin-treated group. Eight Bestatin-treated cases of genital Paget's disease and 8 control cases were similarly followed up. The survival rate was slightly higher for the Bestatin-treated group, but each group consisted of too small a number of cases to demonstrate the utility of Bestatin. The data obtained up to now only suggest the future potentiality of Bestatin treatment for these types of malignancy. For this reason, the data from phase III studies of Bestatin, including previously reported findings on malignant melanoma, proved Bestatin to be useful in treating stage Ib and II malignant melanoma, but were only suggestive of its potential indication in the treatment of other malignant tumors of the skin. Although Bestatin was administered for long periods (a mean of 21.5 months), only mild gastrointestinal disturbances were observed as adverse reactions to the drug in 4.5% of cases. PMID- 2988460 TI - Glandular extramammary Paget's disease. PMID- 2988459 TI - [Phase I-II study of recombinant interferon gamma]. AB - A phase I-II study of human recombinant interferon gamma (rIFN-gamma) was conducted in patients with various advanced cancer refractory to standard chemotherapies. In the phase I study, seven patients received 14 courses of escalating doses ranging from 2 X 10(6)U/m2 to 64 X 10(6)U/m2 by 1-hour intravenous infusion for 5 consecutive days. The toxicities were high fever with chills, anorexia, occasional nausea and vomiting, elevation of serum GOT, and dose-related leukopenia and neurotoxic symptoms such as heavy fatigue with somnolence or lethargy, both of which were reversible. The pharmacokinetics showed that the peak levels of serum rIFN-gamma activity were dose-related but decreased rapidly to below measurable levels within 6 hours after infusion in patients receiving less than 12 X 10(6)U/m2. Considering these data, the dosage of rIFN-gamma 6 X 10(6) U/m2 by daily intramuscular injection for more than 4 weeks was selected for the early phase II study. There was no partial response out of 11 evaluable patients but a stable condition was observed in 2 cases of renal cell carcinoma and one case each of breast cancer and ovarian cancer. All toxicities seen were similar to those observed in the phase I study, but no tachyphylaxis developed with continued dosage. The antitumor effect of rIFN-gamma remains to be evaluated in a further study employing higher doses. PMID- 2988461 TI - Auto-oxidative damage in cement dermatitis. AB - Oxygen intermediates (OIs) generated by stimulated polymorphonuclear leukocytes (PMNs) are known to induce auto-oxidative tissue damage at the site of inflammation. PMNs from five patients with severe and chronic cement dermatitis generated markedly increased levels of OIs. However, only a slight increase in OI generation by PMNs was observed in cement workers without cement dermatitis. Dapsone, which has recently been shown to decrease OI levels, was found to be clinically effective in the treatment of cement dermatitis in these five patients. After treatment, a significant decrease in OI generation was observed in all patients studied. In skin tissues from the cement workers without cement dermatitis, enhanced superoxide-dismutase (SOD) activities as well as increased OI generation by PMNs were noted. In spite of the greatly increased OI generation by PMNs, the SOD activities in the patients were comparable to those in healthy controls. These findings suggest that the severe skin manifestations in patients with cement dermatitis can partly be explained by a defective capacity for enhancing SOD activity which removes increased PMN-derived OIs and thus prevents subsequent tissue injury by OIs at the site of inflammation. PMID- 2988462 TI - Enzyme activities involved in collagen and proteoglycan degradations in oral mucosas of beige (Chediak-Higashi syndrome) mice. PMID- 2988463 TI - Adrenocortical function after acute carbon monoxide exposure in humans. AB - The effect of acute carbon monoxide (CO) exposure on plasma adrenocorticotrophic hormone (ACTH), renin activity (PRA), aldosterone, and cortisol levels was studied in patients approximately 80-90 min after removal from the scene of exposure. Nine patients had carboxyhemoglobin (COHb) levels below 15%. Seven patients had COHb levels that exceeded 15% (Group II). Plasma ACTH, cortisol, and aldosterone levels were significantly higher in Group II (elevated COHb). Plasma cortisol and aldosterone levels were appropriate for the levels of ACTH and PRA achieved. We concluded that (a) acute carbon monoxide poisoning leads to elevated plasma corticosteroid levels, and (b) the adrenal gland appears to function normally acutely after exposure to carbon monoxide. PMID- 2988464 TI - Prostaglandin (PG) E2 generation by cultured canine synovial fibroblasts exposed to microcrystals containing calcium. AB - Immunoreactive prostaglandin (PG) E2 was released into the ambient medium in a dose dependent fashion when either hydroxyapatite (HA) or calcium pyrophosphate dihydrate (CPPD) crystals were added to canine synovial fibroblasts in tissue culture. PGE2 release peaked 6 to 9 hours after HA or CPPD crystals were added in the presence of serum but at 24 hours if they were added in the presence of lactalbumin hydrolysate. PGE2 release correlated with crystal endocytosis estimated qualitatively by serial phase contrast microscopy and time lapse photography. As postulated previously by others for monosodium urate crystals, prostaglandin production by synovial cells may also be related to the pathogenesis of the destructive arthropathies associated with HA or CPPD crystals. PMID- 2988465 TI - Still's disease associated with Coxsackie infection and haemophagocytic syndrome. AB - At the onset of Still's disease (systemic-onset juvenile arthritis) in a 12-year old girl, serological evidence of Coxsackie B virus infection was found. Two weeks later she developed a haemophagocytic syndrome which was then treated with cytotoxic therapy. Her arthritis is still active six years later. PMID- 2988466 TI - [Digital radiocardioangiography as an auxiliary method for the precise localization of zones of myocardial concentration of 99mTc-Sn(II)-pyrophosphate. Preliminary report]. AB - A new method is introduced for the superposition of 99mTc-pyrophosphate (PiF) myocardial images on the isolated images of the radioangiocardiographic dextrophase and levophase, in order to include cardiac anatomical references for the accurate topographic localization of the PiF concentration areas. Results in two different patients are presented. In one of them, the PiF concentration was evident at the inferoseptal, inferoapical and posterolateral regions of the left ventricle. In the other, the PiF concentration was evident at the same areas than in the former plus the inferoanterior region of the right ventricle. Our project is to validate this procedure by correlating its results with those obtained through other clinical methods that detect right ventricular infarction. PMID- 2988467 TI - Inhibitory effect of GABA on sympathetic neurotransmission in rabbit ear artery. AB - GABA 100 microM inhibited neurogenic vasoconstrictor responses elicited in rabbit ear artery by field stimulation at various frequencies. GABA was ineffective both on resting tone and on noradrenaline (0.05-5 microM)- and high-K+ (24-54 mM) induced tonic contraction. GABA effectiveness against field stimulation-induced vasoconstriction was inversely related to the frequency of stimulation. This action of GABA was mimicked by a selective GABAB agonist, such as baclofen (100 microM), but not by selective GABAA agonists as muscimol (100 microM) and homotaurine (100 microM). The selective GABAB antagonists 5-aminovaleric acid (1 mM) and homotaurine (100 microM) completely suppressed the inhibitory effect of GABA on field stimulation-induced vasoconstrictions. GABA action was partially reversed also by the selective GABAA antagonist picrotoxin (100 microM); however, this drug "per se" determined an increase in amplitude of field stimulation induced contractions which in turn could have determined the minor effect of GABA. As a whole these data suggest that GABA can inhibit sympathetic neurotransmission in rabbit ear artery through the stimulation of a prejunctional receptor of the GABAB subtype. PMID- 2988468 TI - Interference of enantiomers of lofexidine with alpha-adrenoceptors. AB - Some alpha-adrenoceptor-mediated cardiovascular activities and alpha-adrenoceptor binding affinities for (+)- and (-)-lofexidine (Dexlofexidine and Levlofexidine) have been studied in comparison with racemic lofexidine. In pithed normotensive rats, i.v. (-)-lofexidine elicited pressor effects at doses (0.1-30 micrograms/kg), which were approximately 20 times lower than those of the (+) isomer. Both yohimbine and prazosin in selective amounts of 1.0 and 0.1 mg/kg (i.v., -15 min), respectively, attenuated the increase in diastolic pressure induced by (+/-)-, (+)- and (-)-lofexidine, showing the involvement of alpha 1- as well as alpha 2-adrenoceptors in the vasopressor responses. No differences were observed in the sensitivity of the pressor effects of the (+)- and (-) enantiomers to blockade by either yohimbine or prazosin. Following i.v. administration to pentobarbitone-anaesthetized normotensive rats, (-)-lofexidine (0.5-5.0 micrograms/kg) was found about 20 times more effective than the dextrorotatory isomer in decreasing mean arterial pressure and heart rate. The increase in heart rate evoked by electrical stimulation in pithed rats was dose dependently reduced by (+/-)-, (-)- and (+)-lofexidine, the (-)-isomer being about 30 times more potent than the (+)-isomer. Similarly, the electrical stimulation-induced increase in diastolic pressure was also most effectively impaired by the laevorotatory enantiomer of lofexidine. (-)-Lofexidine showed an approximately 9-fold higher affinity than (+)-lofexidine for the alpha 2 adrenoceptor-like binding sites in rat brain membranes identified by [3H] clonidine and was 4 times more potent at displacing [3H]-prazosin from alpha 1 adrenoceptors. It is concluded that the alpha-adrenoceptor activity of lofexidine resides predominantly in the (-)-isomer. The isomeric activity ratio of the enantiomers of lofexidine (about 20-fold) is higher than normally found for other imidazolines. PMID- 2988469 TI - Subcellular distribution of dihydropyridine isothiocyanate binding in guinea-pig ileal smooth muscle. AB - The subcellular distribution of the 3H--2,6-dimethyl-3,5-dicarbomethoxy-4(2 isothiocyano) phenyl-1,4-dihydropyridine (DPSCN) binding to guinea-pig ileal smooth muscle was studied by subcellular fractionation. Initial experiments on subcellular fractionation of 3H-DPSCN-labelled tissues by differential centrifugation showed that there was an excellent correlation between the levels of the label present in a fraction and the plasma membrane marker phosphodiesterase I (r = 0.98) but not between the label and the putative endoplasmic reticulum marker NADPH: cytochrome-c-reductase (r = 0.56) or the inner mitochondrial marker cytochrome-c-oxidase (r = 0.36). Centrifugation of the microsomes on a continuous sucrose density gradient showed an excellent correlation of the migration of the label with phosphodiesterase I activity (r = 0.93) but not with the activities of NADPH: cytochrome-c-reductase (r = 0.66) or cytochrome-c-oxidase (r = 0.44). Treatment of microsomes with digitonin (1 mg/ml) followed by centrifugation on continuous sucrose density gradients increased the weighted mean densities of the phosphodiesterase activity (plasma membrane marker) and the labelling by similar magnitudes (0.04 to 0.06 g/ml). The weighted mean densities of NADPH: cytochrome-c-reductase and the cytochrome-c-oxidase were not altered significantly. It is concluded that in the guinea-pig ileal smooth muscle, DPSCN labels the plasma membrane specifically. PMID- 2988471 TI - Comparison of inhibitory actions of chlorpromazine or its 7,8-dihydroxy and 7,8 dioxo-didesmethyl analogs on DA-sensitive adenylate cyclase and calmodulin activation of phosphodiesterase in rat striatum. AB - Two chlorpromazine analogs, 7,8-diOH- and 7,8-dioxo-didesmethyl-chlorpromazine were compared to chlorpromazine (CPZ) with regard to inhibition of three parameters of enzyme activity in rat striatum: 1) dopamine + GTP-sensitive adenylate cyclase in homogenates; 2) dopamine, GTP, calmodulin and Ca++-sensitive adenylate cyclase in washed particulate fractions; and 3) calmodulin-Ca++ activation of high Km cyclic AMP dependent phosphodiesterase in dialyzed supernatant fractions. Chlorpromazine was clearly the most potent antagonist in all three experimental conditions. The CPZ derivatives displayed greatest potency on the particulate adenylate cyclase and all three drugs were 1 to 2 orders of magnitude more effective as inhibitors of the adenylate cyclase preparations than with the calmodulin-Ca++ phosphodiesterase. PMID- 2988470 TI - Effect of cyclic nucleotide phosphodiesterase regulators on isolated rat uterus: with observations on their mechanism of action. AB - The effect of PDE regulators on isolated rat uterus and their probable mechanism of action was investigated. The potassium chloride-induced contracture was enhanced by imidazole (3.67 X 10(-3)M) and potassium bicarbonate (1.5 X 10(-2)M). In contrast, imidazole (pH adjusted with potassium dihydrogen orthophosphate or HCl) produced a biphasic effect, i.e. initial transient relaxation followed by a sustained contracture. A similar biphasic effect of imidazole was observed in uterine preparations suspended in calcium free K2SO4-Ringer, but in preparations which were allowed to equilibrate to a new base line, it produced sustained contracture. The imidazole-induced increase in contracture appears to be mediated through increased influx of calcium. Phosphodiesterase (PDE) stimulation plays only a minor role, if any. The transient relaxant effect of imidazole may be due to inhibition of thromboxane synthetase and the resultant decrease of intracellular calcium release. The relaxant effect of PDE inhibitors (aminophylline, papaverine, diazoxide), catecholamines and db-cyclic AMP on the K2SO4-Ringer and calcium chloride induced contracture was antagonized by imidazole (pH adjusted with HCl). This inhibitory effect of PDE inhibitors may be mediated through the altered membrane permeability to calcium. PMID- 2988472 TI - Influence of intracerebroventricular phenoxybenzamine on the renal action of intracerebroventricular morphine. AB - Renal function is under regulatory influence of the central nervous system, either through mediation of humoral agents or by way of renal nerves, with the sympathetic nerve playing the most important role. As it was recently shown that morphine and enkephalin, when administered intracerebroventricularly (icv), affect renal function in the rabbit, indicating that the opiate receptors are also involved in the central regulation of renal function, it was attempted to clarify whether the central adrenergic system participates in the center-mediated renal action of morphine, employing phenoxybenzamine (PBZ). Morphine, 10 micrograms/kg icv, produced profound and long-lasting antidiuresis and antinatriuresis. PBZ, 250 micrograms/kg icv, elicited marked diuresis with natriuresis and kaluresis, along with significant increases of osmolar clearance and free-water reabsorption, and these effects were not related to a transient increase in glomerular filtration. When PBZ was given 10 min after morphine, the morphine antidiuresis immediately disappeared and the renal action of PBZ unfolded itself undiminished. Also, when morphine was given after PBZ, the PBZ diuresis developed fully, unaffected by morphine. These observations indicate that the central adrenergic system is involved in the regulation of renal function through central opiate receptors. PMID- 2988473 TI - Amiodarone-induced hypothyroidism. A common complication of prolonged therapy: a report of eight cases. AB - Amiodarone is a widely used antiarrhythmic drug, which contains 75 mg of iodide per 200 mg of active substance. Eight of our patients receiving long-term amiodarone therapy became hypothyroid. Seven of these patients had no previous history of thyroid dysfunction or goiter. Antithyroid antibodies were absent, and standard perchlorate discharge tests were positive in seven patients when hypothyroidism was diagnosed. In one patient, amiodarone therapy was withdrawn; over the next nine months, the hypothyroidism resolved, and results of the perchlorate discharge test reverted to normal. We conclude that amiodarone induced hypothyroidism is similar to previously described iodide-induced hypothyroidism. It may develop in the absence of a previous history of thyroid disease, and all patients receiving long-term amiodarone therapy should therefore be regularly monitored for hypothyroidism. PMID- 2988475 TI - [A sensitive method for virus demonstration using Dukol UD]. PMID- 2988474 TI - Successful chemotherapy for cystosarcoma phyllodes in a young woman. AB - Cystosarcoma phyllodes is a rare neoplasm of the breast. The tumor contains both stromal and epithelial elements. Local excision or simple mastectomy is usually curative. Most recurrences are local and can be controlled with excision. We treated a 23-year-old woman in whom disease had metastasized to the lungs with six courses of doxorubicin hydrochloride and cisplatin at three-week intervals and achieved complete remission. PMID- 2988476 TI - [Chromosomal pathology. Recent advances]. PMID- 2988477 TI - [Molecular approach to the action of vitamin D in man]. AB - Some applications to man of specific markers of the molecular action of vitamin D (1.25(OH)2D3 receptors and antibodies to hormone-dependent proteins (CaBP and cDNA] are reported in this study. On case of type II vitamin-dependent rickets was characterized by 1.25(OH)2D3 plasma level greater than 250 pg/ml and a ten fold decrease of the number of binding sites of the hormone in cultured skin fibroblasts. We propose that CaBP 28K and/or 9K-containing cells, such as Purkinje's cells and chondroblasts may be targets for vitamin D action. Detection in fetuses, from the 20th week of gestation, of CaBP 9K messenger RNA in the duodenum and sternum and presence of CaBP 28K and 9K in the chondroblasts of the upper extremity of tibia, suggest that vitamin D acts on the nucleus of its target-cells during fetal development. Finally, discovery of the gene of CaBP 9K in man opens the prospect of studies which will improve the understanding of the mechanism of action of vitamin D. PMID- 2988478 TI - Diagnosis-related groups and clinical research in psychiatry. PMID- 2988479 TI - Electron microscopic assessment of cervical punch biopsies in women followed-up for human papillomavirus (HPV) lesions. AB - Cervical punch biopsies from 102 women prospectively followed-up for HPV (Human papillomavirus) lesions (flat, inverted and papillomatous condylomas) were assessed on transmission electron microscopy (TEM) for the presence of HPV particles. The virus discovery rate was correlated with the other characteristics of significance to the clinical behaviour (natural history) of these lesions, i.e. the colposcopic pattern, expression of papillomavirus genus specific structural antigens (demonstrated by an indirect immunoperoxidase, IP-PAP technique), as well as the degree of HPV-associated cervical intraepithelial neoplasia (CIN). Virus particles were disclosed with equal frequency in all three types of HPV lesions. Noteworthy, and so far unreported, was the discovery of HPV particles in 56 and 64% of the biopsies derived from the epithelium without colposcopic or histological evidence (cytopathic effects) of HPV lesions, respectively. The degree of CIN was without influence on the virus discovery rates. Viral particles were present with equal frequency (57 and 60%, respectively) in the lesions expressing HPV antigens and in those which did not. The highest frequency (89%) of viral particles was found in lesions presented with warty or mosaicism pattern in colposcopy. The implications of these findings to understanding the biology of HPV infections in the uterine cervix is discussed, and the importance of TEM in the follow-up of these lesions is emphasized. PMID- 2988480 TI - Membrane enzyme cytochemical changes in the hepatocytes of diethylnitrosamine induced hepatomas. AB - Male Wistar rats were treated by 9 i.p. injections with 100 mg/kg body weight diethylnitrosamine (DEN), in 9 consecutive weeks. After decapitation, between the 130th and 140th day, primary hepatocellular carcinomas were found. Tissue from 3 different parts of the liver was processed to study the activity of Glucose-6 Phosphatase in the membranes of the endoplasmic reticulum, Adenosinetriphosphatase in the cell (plasma) membrane and of Succinic Dehydrogenase in mitochondrial membranes. In the experimental animals liver tissue revealed no activity of G-6-P in any region affected by the tumor. ATP and SDG activity was dependent on the type of lesion. Generally speaking it diminished with the lesser differentiation of the hepatoma cells. Other morphological and metabolic aspects of liver DEN cancerogenesis are discussed. PMID- 2988481 TI - Perioperative allogeneic blood transfusions. Survival in patients with resected carcinomas of the colon and rectum. AB - Blood transfusion (BT) is reported to cause immunosuppression. We postulated that BT might therefore adversely affect the prognosis of carcinomas of the colon and rectum. We analyzed the overall and recurrence-free survival of 366 patients whose colorectal carcinomas were resected. The 199 patients who received perioperative BTs appeared to survive less well than the 167 patients who received no blood products. However, Cox analysis, which adjusts for other prognostic variables, shows that the difference was not statistically significant. Although 43% of transfused patients survived, as compared with 56.5% of nontransfused patients, this difference was due to variables other than transfusion. This study did not support the hypothesis that BT has an adverse effect on survival of patients with colorectal cancer. PMID- 2988483 TI - [The rearing of female twin cattle using only feed pellets in comparison with straw pellets plus a concentrate mixture. 2. Weight gain and feed consumption as well as rumen fermentation in the 1st half of the 2d year of life]. AB - In investigations with 30 young cows (twin cattle) the applicability of straw materials as sole roughage was tested during the first half of their second year of life (183 feeding days). In the course of the experiment sole feed pellets (I) and partly pelleted rations of dried feed (straw pellets plus loose wheat and loose wheat straw resp.; II) were used. Approximately 72% of the dry matter intake was provided from straw materials in both groups and on an average of the test period. Weight gain and feed expenditure were ascertained and at the end of the test period also the parameters of rumen fermentation and of the concentration of some metabolism parameters in the blood serum. With feed supply being restrictive (2.2 kg dry matter/100 kg live weight), a high dry matter intake from straw (1.6 kg/100 kg live weight or 4.8 kg/animal and day resp.) and average weight gains of between 574 (I) and 582 g (II) were achieved. The standard expenditure of energy (kEFUcattle) and protein stated in the GDR feed evaluation system for this period of development was undercut by 27 (II) and 29 (I)% and 12 (II) and 18 (I)% resp. On average the expenditure of kEFUcattle/kg weight gain was 4.75 (I) and 4.98 (II) and that of digestible crude protein/kg weight gain 679 (I) and 733 g (II). Compared to standard values the expenditure of energy and feed expenditure from concentrates was 80%. Rumen fermentation conditions corresponded to those of a high cellulolytic activity (C2: C3 ratio of greater than 4: 1). The only significant differences between the groups at the time of sampling were to be observed in the total concentration of volatile fatty acids and in the pH-value in the rumen. The absolute (1/animal) and relative (1/kg dry matter intake) volume of rumen fluid varied between 65 and 70 and between 10.5 and 12.5 1. The ascertained concentration of selected metabolism parameters were within the physiologic standard range. PMID- 2988484 TI - [Localization of nucleoside phosphatases in the antral follicle of the guinea pig ovary]. AB - Ultrastructural localization of nucleozidphosphatases (5'-nucleotidase, adenosin triphosphatase (ATPase) and beta-glicerophosphatase) in antral follicles of the guinea-pig ovary has been studied. Certain heterogeneity has been found in distribution of the enzymes: the cells in the follicular tunic possess the greatest 5'-nucleotidase and ATPase activity. When 5'-adenosin monophosphate (5' AMP) is used as a substrate, the lead phosphate residue is mainly revealed in the external surface of plasmolemma and as "caps" in the margical zone of nucleoplasm. ATPase activity is chiefly observed in nucleoli of granular cells and in those of the external follicular tunic cells. Histochemical reaction with 5'-AMP proceeds most intensively in the lucid tunic and in processes of the granular cells contacting with the oocyte. A possibility is discussed on participation of the metabolic enzymes, that localize in these structures, in the mechanisms controlling the oocyte maturation. PMID- 2988482 TI - Results of conservative operations for breast cancer. AB - The results of conservative operations for breast cancer in 1,593 patients treated at the Cleveland Clinic between 1957 through 1975 are reported. During this period, we individualized our treatment of breast cancer depending on tumor size, location in the breast, and clinical stage of the disease. The following three principal operations were performed: modified radical mastectomy in 592 patients (37%), simple (total) mastectomy in 442 patients (28%), and partial (segmental) mastectomy in 291 patients (18%). Survival results at 5, 10, and 15 years are reported. Factors important in long-term survival included stage of the disease, number of lymph node metastases, delay in therapy, size of the tumor, histologic type, and estrogen receptor status; type of operation was not a significant factor. In this series, partial (segmental) mastectomy without radiation therapy provided five- to 15-year survival rates equal to modified radical mastectomy and simple (total) mastectomy. PMID- 2988485 TI - [Morphology and histogenesis problems in cervical cancer]. AB - Both most common and rare varieties of uterine cervix carcinoma were studied light and electron microscopically. It is shown that tumours histologically classified as squamous-cell carcinoma originate either from the squamous epithelium of the ectocervix or from the metaplastic epithelium. Characteristic ultrastructural features of these histogenetic variants of squamous-cell carcinoma are described. The authors' and literature data are presented indicating that the great histological variety of adenocarcinomas and glandular squamous carcinomas is due to the pluripotential properties of proliferating stem cells capable of forming glandular, solid and squamous-cell structures. The source of clear-cell adenocarcinoma may be not only Gartner's duct but the mullerian epithelium as well. The classification of uterine cervix carcinomas reflecting their histogenesis is proposed. PMID- 2988487 TI - [Signet ring cell cancer of the breast]. AB - A rare case of mammary signet-cell carcinoma which was not diagnosed at life is described. In a woman of 28, an enlargement of the right uterine appendages with pleural and lung metastasis were clinically noted and this was interpreted as ovarian carcinoma with pleural and lung metastasis. Histological examination of ovarian tumour performed during the necropsy allowed one to suggest the metastatic signet-cell carcinoma. No tumours were found macroscopically or histologically in the organs of the alimentary canal. Fibro-cystic mastopathy and no nodes were found macroscopically in the mammary glands. Lobular invasive carcinoma consisting of signet-cell elements containing mucus and carcinomatous lymphangoitis were found histologically. Metastasis to the left axillary, perigastral and pancreatic lymph nodes, lungs, carcinomatosis of pleura were revealed. It is suggested, due to its clinical course and histological structure, to distinguish this form of carcinoma as a separate variety of the lobular invasive carcinoma of the mammary gland. PMID- 2988486 TI - [Collagen of synovial sarcomas (immunomorphological studies)]. AB - Results of an immuno-morphological analysis of the I, II, IV and V collagen types in 7 synovial sarcomas of various location are presented. A fibrillar matrix of synovial sarcomas is represented by a collagen of type I and collagen of type V which is co-distributed with the former. Collagen of type III is found in a smaller quantity and in areas of fibrosarcoma-like structure being included in the largest fibrillar structures. Collagen of type IV is presented in all structural areas of tumours. The greatest amount of the IV type collagen is observed in areas of the epithelioid and hemangiopericytoma-like structure, less in areas of fibrosarcoma-like structure. These results may be used in differential diagnostics. In the walls of fissure-like spaces, only the IV type collagen was found as distinct from vascular tumours. On the basis of the data obtained and analysis of the literature a mesodermal origin of synovial sarcoma is suggested. PMID- 2988488 TI - Prevention of hearing impairment from infection and ototoxic drugs. AB - Infectious diseases are a primary cause of hearing impairment and produce about 25% of profound losses. Of these, one fifth are congenital. The major infections include rubella, cytomegalovirus, measles, pertussis, meningitis, and acute otitis media. Hearing loss from ototoxicity is also observed with a number of drugs, notably the aminoglycosides, loop diuretics, and cisplatin. Preventive measures are defined according to primary, secondary, and tertiary principles. Three principles of prevention are considered: direct action, defined objectives, and the variability of effective prevention according to cause. PMID- 2988490 TI - Intravenous pulse methyl prednisolone in the successful treatment of severe sarcoid polyneuropathy with pulmonary involvement. AB - Severe widespread sarcoid polyneuropathy with associated pulmonary involvement was treated successfully in a 26 year old man using "pulse' methyl prednisolone. After he had failed to respond clinically to a two months course of high-dose oral prednisolone, a regimen of intravenous methyl prednisolone 1 g once a week for eight weeks was instituted, along with 10 mg prednisolone orally daily. Neurological improvement began after the third dose with complete remission achieved after the eighth week. More than 12 months after cessation of treatment he remains in good health with no clinical or laboratory evidence of sarcoidosis. PMID- 2988489 TI - A case of herpes zoster associated encephalitis treated with acyclovir. AB - The case of a 67 year old male who developed severe encephalitis associated with herpes zoster ophthalmicus is described. Encephalitis occurred in the absence of cutaneous dissemination and recovery followed treatment with Acyclovir. PMID- 2988491 TI - An unusual incidence of neurological disease affecting horses during a drought. AB - The clinical, pathological and epidemiological factors were investigated in 12 horses presenting with severe neurological signs. Although the cases involved differing central (n = 1), spinal cord (n = 4) and peripheral nerve (n = 7) deficits in a number of instances, there were similar pathological findings. The possibility of a unifying aetiological factor, such as a toxicosis, is discussed because of the pathological similarities and as the cases appeared during an unusually long dry period. PMID- 2988492 TI - The role of contrast angiography in gastrointestinal bleeding with the advent of technetium labelled red blood cell scans. PMID- 2988493 TI - The airline passenger undergoing withdrawal or overdose from narcotics or other drugs. AB - Inflight passenger problems associated with withdrawal or overdose from narcotics or other drugs have been assessed. The specific categories of common drugs of abuse covered are: opiates, amphetamines and cocaine, central nervous system depressants, marijuana, phencyclidine, LSD and mescaline, and peyote. One or more of these drugs may be abused by persons in all walks of life. Relatively recent newspaper accounts of inflight disturbances related to abuse substances have highlighted the problem. Some case histories are given by the authors along with recommendations for methods of dealing with these problems. PMID- 2988494 TI - Selective association in conditioned stress-induced analgesia: functional differences in interoceptive and exteroceptive sensory pathways. AB - Rats were used to determine whether stress-induced analgesia (SIA) can be produced by conditioning with interoceptive stimuli (LiCl) as with exteroceptive stimuli (footshock). SIA was measured using a tail-flick test. As expected, unavoidable footshock as well as conditioning with footshock produced SIA. In contrast, conditioning with LiCl failed to cause SIA. The findings support the notion of functional differences in neural substrates for conditioning by exteroceptive and interoceptive cue. PMID- 2988495 TI - Changes in cardiac glycogen synthase and phosphorylase activities following stimulation of beta-adrenergic receptors in rats. AB - Following a subcutaneous injection of isoprenaline into rats (5 mg X kg-1 b.w.) the cardiac glycogen stores were depleted by about 90% in less than 15 min. Complete restoration of myocardial glycogen was slow (more than 7-8 hours) despite an elevated glycogen synthase activity. A cardioselective beta-adrenergic receptor blockade (using atenolol) resulted in a complete restoration of glycogen stores in 30 min. The results indicate that the potential of myocardial tissue for glycogenogenesis is great but this capability is obscured by continuous glycogenolysis induced by a long-term activation of phosphorylase. The relative importance of beta-receptor stimulation and actual glycogen level in the control of cardiac synthase is discussed. PMID- 2988497 TI - Immobilization of immunoglobulins on silica surfaces. Stability. AB - The development of new immunosensors based on surface-concentration-measuring devices requires a stable and reproducible immobilization of antibodies on well characterized solid surfaces. We here report on the immobilization of immunoglobulin G (IgG) on chemically modified silica surfaces. Such surfaces may be used in various surface-oriented analytical methods. Reactive groups were introduced to the silica surfaces by chemical-vapour deposition of silane. The surfaces were characterized by ellipsometry, contact-angle measurements and scanning electron microscopy. IgG covalently bound by the use of thiol-disulphide exchange reactions, thereby controlling the maximum number of covalent bonds to the surface, was compared with IgG adsorbed on various silica surfaces. This comparison showed that the covalently bound IgG has a superior stability when the pH was lowered or incubation with detergents, urea or ethylene glycol was carried out. The result was evaluated by ellipsometry, an optical technique that renders possible the quantification of amounts of immobilized IgG. The results outline the possibilities of obtaining a controlled covalent binding of biomolecules to solid surfaces with an optimal stability and biological activity of the immobilized molecules. PMID- 2988496 TI - Effect of beta-adrenergic stimulation on free fatty acid content in subepicardial and subendocardial layers of the dog heart in situ. Separation and assay by capillary gas chromatography. AB - The effects of beta-adrenergic stimulation produced by an infusion of isoproterenol (1 microgram X kg-1 min-1, 30 min) were studied in situ in the anaesthetized dog placed under a total cardiopulmonary bypass. Samples of the subepicardial and the subendocardial layers were homogenized separately prior to the extraction and methylation of free fatty acids (FFA). Gas chromatography on Carbowax 20 M capillary columns was used for the quantitation of myristic (C 14:0), palmitic (C 16:0), palmitoleic (C 16:1), stearic (C 18:0), oleic (C 18:1), linoleic (C 18:2), and arachidonic (C 20:4) acids. Within 5 min, isoproterenol decreased the tissue content of FFA significantly. The decrease was more pronounced in the endocardial layer where the FFA concentration reached its minimum at the 5th or the 15th min. In the epicardial layer, all the FFA reached their minimal concentration at the 30th min of the isoproterenol infusion. In both layers, lactate content remained unchanged at 5 and 15 min and rose at the 30th min only and content in phosphorylated compounds (ATP, creatine-phosphate CP) did not show any significant variation during the beta-stimulation period. A significant correlation was found between the chronotropic effect of isoproterenol and the reduction of FFA concentration. PMID- 2988498 TI - Immobilization of immunoglobulins on silica surfaces. Kinetics of immobilization and influence of ionic strength. AB - The kinetics of, and the influence of ionic strength on, the immobilization of rabbit immunoglobulin G (IgG) on different types of well-characterized silica surfaces were investigated. Adsorptive immobilization was compared with covalent attachment via thiol-disulphide exchange reactions. The amount of immobilized IgG on five different types of silica surfaces as a function of IgG concentration, at two different ionic strengths, was determined. The IgG-solid-surface interaction involved different types of interaction forces, depending on the surface chemistry of the solid surface. The solid-surface chemistry is an important parameter determining the immobilized amount of IgG. When conditions for covalent attachment of IgG to the surfaces were fulfilled, the IgG showed high affinity and the immobilized amount of IgG showed a fast saturation. Changes in ionic strength showed no significant influence on the kinetics of immobilization on these surfaces. The amount of covalently attached IgG was partially ionic strength-dependent, indicating that adsorptive interactions were involved. The results are of fundamental interest for the development of new immunosensors based on surface-concentration-measuring devices. PMID- 2988499 TI - Opiates and cultured neuroblastoma x glioma cells. Effect on cyclic AMP and polyamine levels and on ornithine decarboxylase and protein kinase activities. AB - In cultured NG 108-15 neuroblastoma x glioma cells, opiates decreased cellular cyclic AMP and polyamine levels. This decrease was related to the inhibition of ornithine decarboxylase and cyclic AMP-dependent protein kinase activities during the acute exposure of the cells to the drugs. Growing the cells in the presence of opiates for several days led to drug addiction. In the tolerant-addicted cells, polyamine and cyclic AMP levels were close to normal values as were the activities of ornithine decarboxylase and cyclic AMP-dependent protein kinase. Removal of the opiate from 'addicted' cells, by either washing or by adding the antagonist naloxone, resulted in an increase in cyclic AMP and polyamine levels and the activities of ornithine decarboxylase and cyclic AMP-dependent protein kinase. The effect of opiates was closely related to their biological activities. Inactive enantiomorphs did not affect cyclic AMP or polyamine levels; neither did they decrease ornithine decarboxylase and cyclic AMP-dependent protein kinase activities. PMID- 2988501 TI - Effect of glucose on polyphosphoinositide metabolism in isolated rat islets of Langerhans. AB - The metabolism of inositol-containing phospholipids during insulin secretion was studied in rat islets of Langerhans preincubated with [3H]inositol to label their phospholipids. Glucose (20 mM) caused a rapid breakdown of phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-phosphate and an accumulation of inositol trisphosphate and inositol bisphosphate. This effect was maximal at 60s, did not require the presence of extracellular Ca2+, and was abolished by mannoheptulose (15 mM), but not by noradrenaline (1 microM). Mannose (20 mM) and DL-glyceraldehyde (10 mM) produced similar effects to those of glucose, but galactose (20 mM) and KCl (30 mM) were without effect. These results are compatible with the hypothesis that an early event in the stimulus-secretion coupling mechanism in the pancreatic B-cell is the rapid breakdown of polyphosphoinositides catalysed by phospholipase C. Moreover, they suggest that the breakdown of polyphosphoinositides is linked to sugar metabolism in the B cell. This observation is important, since it demonstrates that events in a cell other than plasma-membrane receptor occupancy can promote polyphosphoinositide hydrolysis. PMID- 2988500 TI - Analysis of thyrotropin receptors by photoaffinity labelling. Orientation of receptor subunits in the cell membrane. AB - Porcine thyrotropin (TSH) receptors have been purified by Sepharose-TSH affinity chromatography and crosslinked to a 125I-labelled photoactive derivative (N hydroxysuccinimidyl 4-azidobenzoate; HSAB) of TSH (125I-HSAB-TSH). Purification of the crosslinked complexes on Sephacryl S-300 followed by polyacrylamide-gel electrophoresis in sodium dodecyl sulphate showed that the receptor contained two subunits. One subunit (A) with Mr 45 000 was crosslinked to TSH and the other (B) subunit, Mr 25 000, was linked to the A subunit by a disulphide bridge(s). Other, as yet unidentified, subunits may have been non-covalently associated with the A and B subunits. Analysis of reduced and non-reduced crosslinked TSH receptor-125I HSAB-TSH on Sephacryl S-300 in the presence and absence of detergent indicated that the A subunit was a hydrophilic peptide. This was confirmed in studies of the release into aqueous solution by reducing agent treatment of 125I-HSAB-TSH crosslinked to the TSH receptor A subunit in thyroid membranes. Similar results were obtained with TSH receptors in human thyroid and guinea pig fat cell membranes. These studies suggest that the hydrophilic A subunit of the receptor forms a binding site for TSH on the outside surface of the cell membrane and that the A subunit is linked to the cell membrane by way of a disulphide bridge to the receptor B subunit. PMID- 2988502 TI - Effect of tryptophan metabolites on the activities of rat liver pyridoxal kinase and pyridoxamine 5-phosphate oxidase in vitro. AB - Pyridoxal kinase was purified 4760-fold from rat liver. The Km values for pyridoxine and pyridoxal were 120 and 190 microM respectively, and pyridoxine showed substrate inhibition at above 200 microM. Pyridoxamine 5-phosphate oxidase was also purified 2030-fold from rat liver, and its Km values for pyridoxine 5 phosphate and pyridoxamine 5-phosphate were 0.92 and 1.0 microM respectively. Pyridoxine 5-phosphate gave a maximum velocity that was 5.6-fold greater than with pyridoxamine 5-phosphate and showed strong substrate inhibition at above 6 microM. Among the tryptophan metabolites, picolinate, xanthurenate, quinolinate, tryptamine and 5-hydroxytryptamine inhibited pyridoxal kinase. However, pyridoxamine 5-phosphate oxidase could not be inhibited by tryptophan metabolites, and on the contrary it was activated by 3-hydroxykynurenine and 3 hydroxyanthranilate. Regarding the metabolism of vitamin B-6 in the liver, the effects of tryptophan metabolites that were accumulated in vitamin B-6-deficient rats after tryptophan injection were discussed. PMID- 2988503 TI - The mechanism of end-organ resistance to 1 alpha,25-dihydroxycholecalciferol in the common marmoset. AB - The common marmoset, a New World monkey, requires a large amount of cholecalciferol (110 i.u./day per 100g body wt.) to maintain its normal growth. In a previous report, we demonstrated that the circulating levels of 1 alpha, 25 dihydroxycholecalciferol [1 alpha,25(OH)2D3] in the marmosets are much higher than those in rhesus monkeys and humans, but the marmosets are not hypercalcaemic [Shinki, Shiina, Takahashi, Tanioka, Koizumi & Suda (1983) Biochem. Biophys. Res. Commun. 14, 452-457]. To compare the effect of the daily intake of cholecalciferol, two rhesus monkeys were given a large amount of cholecalciferol (900 i.u./day per 100g body wt). Their serum levels of calcium, 25 hydroxycholecalciferol and 24R,25-dihydroxycholecalciferol were markedly elevated, but the serum 1 alpha,25(OH)2D3 levels remained within a range similar to those in the rhesus monkeys fed the normal diet (intake of cholecalciferol 5 i.u./day per 100g body wt). Intestinal cytosols prepared from both monkeys contained similar 3.5 S macromolecules to which 1 alpha,25(OH)2D3 was bound specifically. However, the cytosols from the marmosets contained only one-sixth as many 1 alpha,25(OH)2D3 receptors as those from the rhesus monkeys. Furthermore, the activity of the 1 alpha,25(OH)2D3-receptor complex in binding to DNA-cellulose was very low in the marmosets. These results suggest that the marmoset possesses an end-organ resistance to 1 alpha,25(OH)2D3 and is a useful animal model for studying the mechanism of vitamin D-dependent rickets, type II. PMID- 2988504 TI - The amino acid sequence of the cytochrome c-554(547) from the chemolithotrophic bacterium Thiobacillus neapolitanus. AB - An amino acid sequence is proposed for the cytochrome c-554(547) from the bacterium Thiobacillus neapolitanus N.C.I.B. 8539). It consists of a polypeptide chain of 91 residues, with a pair of haem-attachment cysteine residues at positions 15 and 18. There is similarity in sequence with each of the halves of the sequence of the dihaem cytochromes c4 and with a cytochrome c-554(548) from a halophilic strain of Paracoccus. Detailed evidence for the amino acid sequence of the protein has been deposited as Supplementary Publication SUP 50127 (11 pages) at the British Library (Lending Division), Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1985) 225, 5. PMID- 2988505 TI - Substrates for cyclic AMP-dependent protein kinase in islets of Langerhans. Studies with forskolin and catalytic subunit. AB - To investigate substrates for cyclic AMP-dependent protein kinase in intact islets of Langerhans, batches of islets were incubated with [32P]Pi for 1 h in the presence of 10 mM-glucose; the adenylate cyclase activator forskolin, which in parallel experiments was shown to increase islet cyclic AMP content and insulin release, was then added. Islets were homogenized and subcellular fractions prepared by differential centrifugation. Phosphopeptides were electrophoresed on sodium dodecyl sulphate/polyacrylamide gels and quantified by autoradiography and densitometry. Within 5 min forskolin caused increased labelling of Mr-25 000 and -30 000 cytosolic and Mr-23 000 and -32 000 particulate peptides; a rapid decrease in phosphorylation of Mr-18 000 and -34 000 cytosolic peptides was also observed. In addition, rather slower phosphorylation occurred of the Mr-15 000 peptide previously identified as histone H3 [Christie & Ashcroft (1984) Biochem. J. 218, 87-99]. When similar subcellular fractions were incubated with [gamma-32P]ATP and purified catalytic subunit of cyclic AMP-dependent protein kinase, peptides phosphorylated included cytosolic species of Mr 25 000 and 30 000 and particulate species of Mr 23 000 and 32 000. The distribution of RNA in the subcellular fractions suggested that the Mr-32 000 species could be a ribosomal protein. The 24 000 g pellet was heterogeneous, as judged by marker assays, and was therefore fractionated further by Percoll-density-gradient centrifugation. The peak containing the Mr-23 000 peptide was resolved from marker enzymes for plasma membranes, mitochondria and endoplasmic reticulum and coincided with a peak for insulin: hence the Mr-23 000 peptide is likely to be a secretory-granule component. The study demonstrates that the potentiation of insulin release that occurs when islet cyclic AMP is increased is accompanied by rapid phosphorylation of specific islet substrates for cyclic AMP-dependent protein kinase. The data are consistent with the hypothesis that protein phosphorylation is involved in the regulation of insulin secretion. PMID- 2988506 TI - The action of adenosine in relation to that of insulin on the low-Km cyclic AMP phosphodiesterase in rat adipocytes. AB - The adenosine-sensitive cyclic AMP phosphodiesterase of rat adipocytes was found to reside in the same subcellular fraction as the enzyme sensitive to insulin. There were several similarities between the action of adenosine and that of insulin on the enzyme. The action of adenosine on the phosphodiesterase is probably like that of insulin, both being receptor-mediated, although different sites or different receptors could be involved. Adenosine analogues with intact ribose but a modified purine moiety elicited a response similar to that of adenosine. Added Ca2+ was also not a requirement for the action of adenosine. The action of adenosine was not synergistic with that of insulin, neither was adenosine essential for insulin action. Insulin stimulated the enzyme even at low cell concentrations and in the presence of adenosine deaminase. Adenosine, however, enhanced the effect of insulin, but only at insulin concentrations that produced submaximal effects. Thus the mechanisms of action could be similar or related. The time-course effect of a suboptimal concentration of insulin was transitory, like that of adenosine, and was influenced by the presence of adenosine, whereas that of a maximally effective concentration of insulin was sustained for at least 20 min and was not affected by the presence of adenosine. Isoprenaline enhanced phosphodiesterase activity stimulated by optimal concentrations of either adenosine or insulin, suggesting that their effects were mediated through different mechanisms of action. PMID- 2988507 TI - Identification of a cellular 110 000-Da protein substrate for the insulin receptor kinase. AB - Addition of insulin to wheat-germ-lectin-purified glycoproteins derived from rat hepatocytes or rabbit brown adipose tissue results in the increased phosphorylation of a Mr-110 000 protein. This naturally occurring glycoprotein appears as a monomeric structure and is not part of the insulin receptor itself, since it is not immunoprecipitated by highly specific antibodies to insulin receptor. Phosphorylation of the Mr-110 000 protein and autophosphorylation of the receptor beta-subunit (Mr 95 000) are stimulated by insulin in a remarkably similar dose-dependent fasion, with half-maximal stimulation at 1 nM-insulin. Further, kinetic studies suggest that the phosphorylation of the Mr-110 000 protein occurs after autophosphorylation of the insulin-receptor kinase. In conclusion, the present identification of an endogenous substrate for the insulin receptor kinase could suggest that some, if not all, effects of insulin may be mediated through activation of this kinase. PMID- 2988508 TI - Complexes with halide and other anions of the molybdenum centre of nitrate reductase from Escherichia coli. AB - The interconversion of nitrate reductase from Escherichia coli between low-pH and high-pH Mo(V) e.p.r. signal-giving species was re-investigated [cf. Vincent & Bray (1978) Biochem. J. 171, 639-647]. The process cannot be described by a single pK value, since the apparent pK for interconversion is raised by the presence of various anions. The low-pH form of the enzyme exists as a series of complexes with different anion ligands of molybdenum. Each complex has specific and slightly different e.p.r. parameters, but all show strong coupling of Mo(V) to a single proton, exchangeable with the solvent, having A(1H)av. 1.0 to 1.3 mT. Complexes with Cl-, F- [A(19F)av. 0.7 mT], NO3- and NO2- give particularly well defined spectra. The high-pH form of the enzyme is now shown to bear a coupled proton. Like that in the low-pH species, this proton is exchangeable with the solvent, but the coupling is much weaker, with A(1H)av. 0.3 mT. Thus, contrary to earlier assumptions, the proton detectable by e.p.r. is probably not identical with the proton whose dissociation controls interconversion between the two species; the latter proton could be located in the protein rather than on a ligand of molybdenum. Treatment of the enzyme with trypsin [Morpeth & Boxer (1985) Biochemistry 24, 40-46] did not affect its Mo(V) e.p.r. signals. PMID- 2988509 TI - Pertussis toxin does not inhibit muscarinic-receptor-mediated phosphoinositide hydrolysis or calcium mobilization. AB - Pertussis toxin was used to examine the role of the inhibitory guanine nucleotide regulatory protein, Ni, in muscarinic-receptor-mediated stimulation of phosphoinositide turnover and calcium mobilization. In cultured chick heart cells, pertussis-toxin treatment inhibited muscarinic-receptor-mediated attenuation of isoprenaline-stimulated cyclic AMP accumulation. This finding is consistent with the proposal that pertussis toxin blocks the capacity of Ni to couple muscarinic receptors to adenylate cyclase. In contrast, treatment of chick heart cells or 1321N1 human astrocytoma cells with pertussis toxin did not block muscarinic-receptor-mediated stimulation of phosphoinositide hydrolysis, as measured by [3H]inositol phosphate accumulation in the presence of Li+. Pertussis toxin treatment also had little effect on basal and muscarinic-receptor stimulated phosphatidylinositol synthesis, as measured by the incorporation of [3H]inositol into phosphatidylinositol. Activation of muscarinic receptors also enhances the rate of unidirectional 45Ca2+ efflux in 1321N1 cells; this response, like phosphoinositide hydrolysis, was not prevented by pertussis-toxin treatment. Our data suggest that muscarinic receptors are not coupled to phosphoinositide hydrolysis or calcium mobilization through Ni. PMID- 2988510 TI - Breakdown of phosphatidylinositol 4,5-bisphosphate in a T-cell leukaemia line stimulated by phytohaemagglutinin is not dependent on Ca2+ mobilization. AB - Addition of phytohaemagglutinin (PHA) to the [32P]Pi-prelabelled JURKAT cells, a human T-cell leukaemia line, resulted in a decrease of [32P]phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] to about 35% of the control value. The decrease was almost complete within 30s after the PHA addition. This decrease was followed by an increase in the 32P-labelling of phosphatidic acid (maximally 2.8-fold at 2 min). The stimulation of myo-[2-3H]inositol-prelabelled JURKAT cells by PHA induced an accumulation of [2-3H]inositol trisphosphate in the presence of 5 mM LiCl. The result indicates hydrolysis of PtdIns (4,5)P2 by a phospholipase C. The PHA stimulation of JURKAT cells induced about 6-fold increase in the cytosolic free Ca2+ concentration, [Ca2+]i, which was reported by Quin-2, a fluorescent Ca2+ indicator. Studies with partially Ca2+-depleted JURKAT cells, with the Ca2+ ionophore A23187, and with 8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate indicate that the breakdown of PtdIns(4,5)P2 is not mediated through changes of [Ca2+]i. These results therefore indicate that the PHA-induced breakdown of PtdIns(4,5)P2 in JURKAT cells is not dependent on the Ca2+ mobilization. PMID- 2988511 TI - Basal phosphorylation of cyclic AMP-regulated phosphoproteins in intact S49 mouse lymphoma cells. AB - Protein phosphorylation in intact S49 mouse lymphoma cells was studied by using high-resolution two-dimensional gel electrophoresis of proteins labelled with [35S]methionine or [32P]Pi. In wild-type cells substrates for cyclic AMP stimulatable phosphorylation exhibited high basal phosphorylation; in mutant cells deficient in activities of either cyclic AMP-dependent protein kinase or adenylate cyclase, basal phosphorylation of most of these substrates was negligible. Analysis of tryptic phosphopeptides from proteins labelled with [32P]Pi in wild-type cells suggested that identical sites were phosphorylated under conditions of both basal and hormonally elevated concentrations of cyclic AMP. These results argue that most basal phosphorylation is a consequence of partial activation of cyclic AMP-dependent protein kinase and that this activation is attributable to basal concentrations of cyclic AMP. For the intermediate filament protein vimentin, basal phosphorylation was largely at a site distinct from that stimulated by increased cyclic AMP, and basal phosphorylation was not markedly different in mutant and wild-type cells. Vimentin phosphorylated at both sites was not observed. Cyclic AMP treatment resulted in enhanced phosphorylation at the cyclic AMP-specific site and decreased phosphorylation at the cyclic AMP-independent site. PMID- 2988512 TI - Interaction of autoantibodies to thyrotropin receptor with a hydrophilic subunit of the thyrotropin receptor. AB - Reduction of human thyroid membranes with dithiothreitol caused the release of a water-soluble glycoprotein which neutralized the thyrotropin (TSH) receptor binding and thyroid-stimulating activities of Graves' serum. Analysis of the protein by gel filtration and sucrose density gradient centrifugation allowed estimates of 3.45 nm for the Stokes' radius, 3.6 S for the s20,w and 47 000 +/- 5000 (mean +/- S.D.; n = 4) for the Mr. The material released by dithiothreitol treatment could be crosslinked to 125I-labelled TSH coupled to N hydroxysuccinimidyl 4-azidobenzoate (125I-HSAB-TSH), suggesting that it contained a component of the TSH receptor. Furthermore, analysis of the crosslinked material by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicated that it contained the TSH receptor A subunit (Mr 50 000). Several factors suggested therefore that the glycoprotein released by dithiothreitol treatment of human thyroid membranes was the TSH receptor A subunit. In particular, (a) both preparations were hydrophilic and were released from membranes by reduction, (b) they had similar Mr values and (c) both preparations crosslinked to 125I-HSAB TSH. Material similar to the TSH receptor A subunit was released from thyroid membranes by treatment with papain, probably as a result of cleavage of the receptor A subunit at a site close to the interchain disulphide bridge. A similar mechanism, involving thyroid proteinases, was probably involved in release of material with similar properties to the TSH receptor A subunit during freezing and thawing of human thyroid homogenates. PMID- 2988513 TI - Molecular modelling of human complement subcomponent C1q and its complex with C1r2C1s2 derived from neutron-scattering curves and hydrodynamic properties. AB - Models for the structures of subcomponent C1q of first component C1 of human complement and its complex with subunit C1r2C1s2 are compared with experimental neutron-scattering curves. The length of the C1q collagenous arm is closer to 14.5 nm than to 11.5 nm proposed from electron microscopy, and this is consistent with the primary sequence of C1q. The mean C1q base-arm angle is 40-45 degrees and C1q is found to be flexible: the base-arm angle can vary up to 30 degrees from equilibrium at any moment. The complex of C1r2C1s2 and C1q requires a large shape change in C1r2C1s2. Ring-like models for C1r2C1s2 are not as successful at rationalizing the scattering data as are models that involve C1r2C1s2 binding to one side of C1q. Hydrodynamic calculations of the sedimentation coefficients for C1q and C1 are generally consistent with these neutron models. PMID- 2988514 TI - Effects of detergents and cytochrome c binding on scalar and vectorial proton ejection by proteoliposomes containing cytochrome oxidase. AB - The detergent lauryl maltoside abolishes respiratory control and proton ejection by cytochrome c oxidase-containing proteoliposomes over a narrow concentration range. Expression of cryptic activity (inward-facing oxidase) is released over the same concentration range. Catalytic functions (Vmax. and Km) of the enzyme are not changed by the detergent. Lipid micelles containing detergent bind approximately the same amount of cytochrome c as do vesicles containing an equivalent amount of lipid. Uncoupler-insensitive proton release is seen when proteoliposomes are pulsed with ferrocytochrome c at low ionic strength. Such uncoupler-insensitive acidification is not seen at higher ionic strength, nor with oxygen pulses of anaerobic solutions previously incubated with cytochrome c. Vesicles at low ionic strength catalyse cytochrome c autoxidation; this process can mimic proton re-equilibration in systems that have pumped protons from inside to the bulk phase. Proton re-equilibration following a pulse of cytochrome c or oxygen is multiphasic. The slowest phases are attributed to vesicle heterogeneity, some internal alkali being retained within vesicles of low intrinsic proton permeability. This can be overcome by the addition of either very low levels of carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone or high levels of valinomycin. PMID- 2988515 TI - Physicochemical transfer of [3H]cholesterol from plasma lipoproteins to cultured human fibroblasts. AB - The transfer of free cholesterol from [3H]cholesterol-labelled plasma lipoproteins to cultured human lung fibroblasts was studied in a serum-free medium. The uptake of [3H]cholesterol depended upon time of incubation, concentration of lipoprotein in the medium, and temperature. Modified (reduced and methylated) low-density lipoprotein (LDL), which did not enter the cells by the receptor pathway, gave a somewhat lower transfer rate than unmodified LDL, but if the transfer values for native LDL were corrected for the receptor mediated uptake of cholesterol the difference was eliminated. The initial rates of transfer of [3H]cholesterol from LDL and high-density lipoprotein (HDL) were of the same order of magnitude (0.67 +/- 0.05 and 0.75 +/- 0.06 nmol of cholesterol/h per mg of cell protein, respectively) while that from very-low density lipoprotein (VLDL) was much lower (0.23 +/- 0.02 nmol of cholesterol/h per mg) (means +/- S.D., n = 5). The activation energy for transfer of cholesterol from reduced, methylated LDL to fibroblasts was determined to be 57.5 kJ/mol. If albumin was added to the incubation medium the transfer of [3H]cholesterol was enhanced, while that of [14C]dipalmitoyl phosphatidylcholine was decreased compared with the protein-free system. The results demonstrate that, in spite of its low water solubility, free cholesterol can move from lipoproteins to cellular membranes, probably by aqueous diffusion. We propose that physicochemical transfer of free cholesterol may be a significant mechanism for net uptake of the sterol into the artery during atherogenesis. PMID- 2988516 TI - The beta-lactamase of Enterobacter cloacae P99. Chemical properties, N-terminal sequence and interaction with 6 beta-halogenopenicillanates. AB - The beta-lactamase of Enterobacter cloacae P99 consists of one polypeptide chain of Mr 39000 devoid of disulphide bridges and free thiol groups. It contains an unusually high proportion of tyrosine and tryptophan. The N-terminal sequence exhibits overlaps with the tryptic peptide obtained after labelling the active site with 6 beta-iodopenicillanate. The active-site serine residue is at position 64. The homology with the chromosomal beta-lactamase of Escherichia coli K 12 (ampC gene) is lower within the 25 residues of the N-terminal portion than around the active-site serine residue. The P99 beta-lactamase is inactivated by 6 beta bromo- and 6 beta-iodo-penicillanate, with a second-order rate constant of 110 140M-1 X s-1 at 30 degrees C and pH 7.0, a value that is much lower than that observed with class-A beta-lactamases. PMID- 2988517 TI - 'Non-specific' binding. The problem, and a solution. AB - The concept of 'non-specific' binding, as it relates to studies of the binding of hormones to their receptors, is reviewed. It is concluded that the most widely used operational definition, namely binding that is not displaceable by an excess of unlabelled ligand, is often inaccurate, resulting either in overestimation of the number of high-affinity receptors and underestimation of the affinity of a given hormone for its receptor, or in a curvilinear Scatchard plot suggesting (artifactually) the presence of negative co-operativity or multiple classes of binding sites. The general use of an alternative approach to non-specific binding, in which the non-specific component is assessed from an analysis of total binding, is advocated. The superiority of this approach is illustrated with data on the binding of high-density lipoproteins to their receptors. PMID- 2988519 TI - Regulation of gluconeogenesis by glycerol and its phosphorylated derivatives. AB - Glycerol, glycerol-3-phosphate (G3P), and dihydroxyacetone phosphate (DHAP) were evaluated as inhibitors of gluconeogenesis on rat liver enzymes in vitro, and for their effects on glucose formation in vivo in well-nourished and malnourished rats. DHAP was more potent as an inhibitor than G3P on fructose-1,6-diphosphatase (FDPase), phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pase). The I50 for DHAP was 2, 8, and 9 x 10(-3) M, respectively. No effect was observed on rat liver pyruvate carboxylase (PC). Glycerol was a weak inhibitor of FDPase and PEPCK, but did not inhibit PC and G6Pase. In vivo, when G3P was injected before a parenteral L-alanine (Ala) challenge, it produced a hypoglycemic effect in malnourished rats and a lesser, but noticeable, blood glucose level reduction in well-fed animals. Glycerol caused a smaller reduction in glucose formation from Ala. No comparable effects were observed after a fructose pretreatment. These results underscore the potential hypoglycemic effects of phosphorylated glycerol metabolites and identify the steps in gluconeogenesis where this action is exerted. The study also stresses the nutritional component in the glycerol intolerance syndrome, apparent from the far more severe effects observed in malnourished rats given G3P or glycerol prior to Ala. PMID- 2988518 TI - Intracellular calcium and adenosine 3',5'-cyclic monophosphate as mediators of potassium-induced aldosterone secretion. AB - We compared the action of K+ on aldosterone secretion from isolated bovine adrenal glomerulosa cells with that of ionophore A23187. Addition of either 50 nM A23187 or 8 mM-K+ to perifused cells induces a similar initial aldosterone secretory responses, and a similar sustained increases in Ca2+ entry. However, K+ induced secretion is more sustained than is A23187-induced secretion, even though each agonist appears to act by increasing Ca2+ entry into the cells. When [3H]inositol-labelled cells are stimulated by 8 mM-K+, a small decrease in phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] is observed. This decrease is not accompanied by an increase in inositol trisphosphate (InsP3) concentration. Also, if [3H]arachidonic acid-labelled cells are exposed to 8 mM K+, there is no increase in [3H]diacylglycerol production. When [3H]inositol labelled cells are stimulated by 50 nM-A23187, a small decrease in PtdIns(4,5)P2 is observed. This decrease is not accompanied by an increase in InsP3. The cyclic AMP content of K+-treated cells was approximately twice that in A23187-treated cells. If cells are perifused simultaneously with 50 nM-forskolin and 50 nM A23187, the initial aldosterone-secretory response is similar to that induced by A23187 alone, and the response is sustained rather than transient, and is similar to that seen during perifusion of cells with 8 mM-K+. This dose of forskolin (50 nM) causes an elevation of cyclic AMP concentration in A23187-treated cells, to a value similar to that in K+-treated cells. These results indicate that, in K+ treated cells, a rise in cyclic AMP content serves as a positive sensitivity modulator of the Ca2+ message, and plays a key role in mediating the sustained aldosterone-secretory response. PMID- 2988520 TI - Adrenal dysfunction in glycerol kinase deficiency. AB - The infantile form of glycerol kinase deficiency appears to be an X-linked disorder which is consistently characterized by developmental delay and adrenal cortical insufficiency and hypoplasia. We propose that the inherited deficiency of outer mitochondrial membrane-bound glycerol kinase restricts glycerophospholipid synthesis, and, hence, the activation of steroidogenesis. This would limit the conversion of cholesterol to pregnenolone, the precursor for glucocorticoids in the adrenal cortex. The deficiency in cortisol production, with a lack of feedback to the pituitary, would result in increased ACTH production and hypertrophy of the fascicular zone at the same time that replication of the cells within this zone would be inhibited. Similarly, the decreased mineralocorticoid production by the sparse glomerulosal zone would limit the ability of the individual to respond to stress, and would result in development of potentially fatal hyponatremia and hyperkalemia. Organization of the pathway for glycerophospholipid synthesis at the outer mitochondrial membrane would make this pathway particularly vulnerable to mutations disrupting the compartmented production of the parent compound, glycerol 3-phosphate, by mitochondrial-bound glycerol kinase. PMID- 2988523 TI - Characterization of the DNA of the hamster papovavirus: II. A comparison of binding sites for Escherichia coli- and calf thymus RNA polymerase II on the hamster papovavirus genome. AB - Calf thymus (CT) RNA polymerase II bound to hamster papovavirus (HaPV) DNA was visualized by electron microscopy and compared to binding sites obtained after binding of Escherichia coli RNA polymerase to the HaPV genome. Thirteen binding sites were observed with CT polymerase II at map positions 0.04-0.07; 0.11; 0.18; 0.30; 0.37; 0.47; 0.57; 0.65; 0.78; 0.83; 0.90 and 0.94 using the unique BamHI cleavage site as zero point on the HaPV physical map. These binding sites correlate well with A + T rich sequences within the HaPV DNA as revealed by experiments using protein 32 coded for by phage T4. A comparison of binding of prokaryotic and eukaryotic RNA polymerase demonstrates a high degree of correspondence for most of the binding sites on the HaPV genome. PMID- 2988521 TI - Glycogen storage disease type IB: a new model of genetic disorders involving the transport system of intracellular membrane. AB - Our studies have revealed that the primary lesion of GSD type Ib exists in the G6P transport system in the microsomal membrane. Distinct evidence for the existence of a specific G6P transport system in microsomal membrane was obtained through these studies. This is the first example of a genetic disorder involving the transport system of an intracellular membrane. HHH syndrome (hyperornithinemia, hyperammonemia, and homocitrullinuria), in which the transport of ornithine to the mitochondria is presumed to be defective, may be another example belonging to this category of genetic disorders (18-20). A possibility exists that there are many other disorders due to defects in the membrane transport of intracellular organelles. PMID- 2988524 TI - Fluoride ions increase collagenase production by rabbit synovial fibroblasts. AB - Ionic fluoride is taken up by rabbit synovial fibroblasts in culture. The uptake was accompanied by an increased production of latent collagenase, and decreased activity of 5'-nucleotidase. PMID- 2988522 TI - The regulation of mitochondrial-bound hexokinases in the liver. AB - A functional coupling between bound hexokinase and the inner mitochondrial compartment has been shown. It is based structurally on the binding of hexokinase to a pore protein which is present in zones of contact between the two boundary membranes. The latter was observed by electron microscopic localization of antiporin and hexokinase at the mitochondrial surface. The four isoenzymes present in liver differ considerably in their activity after binding to the mitochondrial surface. This was found by binding studies using the four isoenzymes isolated from the supernatant. Isoenzyme IV did not bind at all. Isoenzymes I-III did bind and became activated: I, 5.9-fold; II, 39-fold; and III, 1.3-fold. These results suggest that the in vivo activity of hexokinase in the mitochondrial fraction is much larger than so far observed. Furthermore the binding of isoenzymes was differently affected by metabolites. Glucose-6 phosphate exclusively desorbed isoenzyme I from the mitochondrial membrane whereas free fatty acids predominantly liberated isoenzymes II and III. A reciprocal change of the levels of free fatty acids and glucose 6-phosphate in livers of starved rats therefore, can explain why exclusively mitochondrial-bound isoenzymes II and III decreased 10-fold while at the same time isoenzyme I increased. PMID- 2988525 TI - Dopamine-stimulated fucosylation of brain proteins in vitro is not inhibited by puromycin. AB - In slices of rat hippocampus, the influence of puromycin (0.5 mM) on dopamine (0.5 mM)-stimulated L-fucose incorporation into glycoproteins was studied. Puromycin (15 min, 2 h and 4 h pretreatment) inhibited protein synthesis by some 70% (estimated by the in vitro incorporation rate of L-leucine), whilst L-fucose incorporation into rat hippocampal glycoproteins was only inhibited about -20% (4 h pretreatment) compared to control values (100%). Under these experimental conditions, the dopamine-induced increase in both the L-fucose incorporation into glycoproteins and the activity of fucokinase was not affected by puromycin. These data suggest that these short-lasting dopamine actions do not depend on protein synthesis. PMID- 2988526 TI - Occurrence of a polyubiquitin structure in ubiquitin-protein conjugates. AB - In the ubiquitin-mediated pathway for the degradation of intracellular proteins, several molecules of ubiquitin are linked to the protein substrate by amide linkages. It was noted that the number of ubiquitin-protein conjugates and their apparent molecular size are higher than expected from the number of amino groups in the protein. When the amino groups of ubiquitin were blocked by reductive methylation, it was efficiently conjugated to lysozyme, but the higher-molecular weight conjugates were not formed. This suggests that the higher-molecular-weight conjugates with native ubiquitin contain structures in which one molecule of ubiquitin is linked to an amino group of another molecule of ubiquitin. Methylated ubiquitin stimulated protein breakdown at about one half the rate obtained with native ubiquitin, and isolated conjugates of 125I-lysozyme with methylated ubiquitin were broken down by reticulocyte extracts. These findings indicate that the formation of polyubiquitin chains is not obligatory for protein breakdown, though it may accelerate the rate of this process. PMID- 2988527 TI - Isolation of S-100 binding proteins from brain by affinity chromatography. AB - S-100-binding proteins, and calmodulin-binding proteins were isolated from S-100- and calmodulin-depleted bovine brain extract by Ca2+-dependent affinity chromatography using S-100- and calmodulin-coupled Sepharose columns respectively. The majority of the protein (80 to 90%) including calcineurin that bound to S-100 also bound to calmodulin and vice versa, suggesting both proteins may regulate common targets. However these two regulatory proteins also bind few other proteins specific for each. These include cyclic nucleotide phosphodiesterase, 55k, and 220k proteins for calmodulin and 24k, 42k, and 90k proteins for S-100. Certain proteins also specifically bound to S-100 both in Ca2+-dependent and independent ways. In glial cells S-100 protein may replace calmodulin in regulating Ca2+-influenced functions. PMID- 2988528 TI - Identification of bovine brain Ca2+-binding proteins. AB - In a previous communication (Waisman, D.M., Smallwood, J.I., Lafreniere, D. and Rasmussen, H. (1983) Biochem, Biophys. Res. Commun. 116, 435-441) we reported that chromatography of bovine brain 100,000 X g supernatant on diethylaminoethyl (DEAE) cellulose and analysis of resultant fractions by chelex competitive calcium binding assay, resolved three peaks of calcium binding activity. Gel permeation chromatographic analysis of each peak resolved apparent Mr 40,000 (Peak I), Mr 75,000, Mr 230,000 and Mr 420,000 (Peak II), and Mr 38,000 (Peak III). In the present communication the calcium binding proteins responsible for the calcium binding activity peaks resolved by gel permeation chromatography, have been purified and identified as caligulin, (Mr 40,000), calcineurin, (Mr 230,000) and calmodulin, (Mr 38,000). In addition, a novel calcium binding protein (Mr 48,000 by SDS PAGE) has been identified from the Mr 75,000 calcium binding activity peak. PMID- 2988529 TI - Low levels of mercury inhibit the respiratory burst in human polymorphonuclear leukocytes. AB - The objective of this investigation was to examine the effects of low levels of Hg(II) on the respiratory burst of PMNs by monitoring O2 consumption, superoxide radical formation, and chemiluminescence. Hg(II) at concentrations of 10-100 ng/ml profoundly inhibited zymosan-stimulated human cells. This inhibition was immediate in onset and occurred with minimal loss of cell viability. Effects of Hg(II) on the PMN respiratory burst were compared with those of Sn, Pb, Se, Au, Ag and Cu. Only in the case of Ag and Cu did the inhibitory effects approach those of Hg. The results indicate that Hg(II) may serve as a specific inhibitor of components of the respiratory burst. PMID- 2988531 TI - Activation of bovine heart ATP-MG2+-dependent phosphoprotein phosphatase: isolation of a phosphoenzyme intermediate and its conversion to the active form via a Mg2+-dependent autodephosphorylation reaction. AB - The ATP-Mg2+-dependent protein phosphatase, a holoenzyme form of type I protein phosphatase (phosphatase-1) requires the action of phosphatase-1 kinase (FA) for activation. The enzyme (75 kDa) purified from bovine heart consists of a catalytic (C) and a regulatory (R) subunit of 40 kDa and 34 kDa, respectively, and activation is associated with phosphorylation of the R-sub-unit. A procedure has been developed for isolation of [32P]phosphatase-1 ( [32P]E-P) in non denatured form. In the absence of divalent cation, [32P]E-P is catalytically inactive and the phosphorylation is stable. Addition of Mg2+ triggers autodephosphorylation of [32P]E-P with concomitant generation of phosphorylase phosphatase activity. The autodephosphorylation/activation process is dependent on Mg2+ concentration. The KA value for Mg2+ is 0.6 mM. The phosphorylase phosphatase activity generated from the release of 1 mol. 32P is 1.1 X 10(12) units which is equivalent to 15,000 units per mg enzyme protein. The present findings provide direct evidence that the phosphorylated phosphatase-1 is not the active form (Ea). Instead, Ea is directly produced from the intermediate by a Mg2+-dependent autodephosphorylation reaction. PMID- 2988530 TI - Is cAMP the obligatory second messenger in the action of lutropin on Leydig cell steroidogenesis. AB - Two adenylate cyclase inhibitors: 9-(tetrahydro-2-furyl)adenine and 2'5' dideoxyadenosine decreased cAMP levels in LH-stimulated immature rat Leydig cells by 20-40%, independent of the concentration of LH. Steroid production was not correlated with this decrease in cAMP, but was increased (146%). The phorbol ester 4 beta-phorbol-12-myristate-13-acetate stimulated steroidogenesis and the phosphorylation of a 17 kD and a 33 kD protein, which was also stimulated by LH, whereas the inactive phorbol ester 4 beta-phorbol-12,13-diacetate did not have any effects. Moreover, the Ca2+-channel blocker diltiazem inhibited LH effects, but had no direct effects on the cholesterol side chain cleavage enzyme. It is concluded that cAMP may not be the only second messenger in LH action, and that other second messenger systems are probably also involved. PMID- 2988532 TI - Modulation of the activity of purified phosphatidylinositol 4-phosphate kinase by phosphorylated and dephosphorylated B-50 protein. AB - To investigate the modulation of phosphatidylinositol 4-phosphate kinase activity by the degree of phosphorylation of the B-50 protein, the enzyme was purified from rat brain cytosol by ammonium sulphate precipitation and DEAE-cellulose column chromatography. Purified rat brain B-50 was phosphorylated with protein kinase C and dephosphorylated with alkaline phosphatase. Incubation of the semi purified phosphatidylinositol 4-phosphate kinase with 1 microgram of the B-50 preparation enriched in the dephospho-form, resulted in a small reduction of phosphatidylinositol 4-phosphate kinase activity (-16%), whereas incubation with the phospho B-50 preparation inhibited the enzyme activity by 40%. The effect of exogenous B-50 was studied in the presence of 10 micrograms albumin to minimize aspecific protein-protein interactions. The present data on the effect of exogenous B-50 protein on phosphatidylinositol 4-phosphate kinase activity, further support our hypothesis that the phosphorylation state of B-50 may be a regulatory factor in phosphoinositide metabolism in rat brain. PMID- 2988533 TI - Beta-endorphin: photoaffinity labelling of a non-opioid binding site in a human neuroblastoma. AB - A photoaffinity reagent 2-nitro-4-azidophenylsulfenyl chloride (2,4-NAPS-Cl) and synthetic analogs of human beta-endorphin (beta h-EP) were employed to demonstrate the presence of receptor sites specific for beta h-EP but of non opioid character in a human neuroblastoma cell line (IMR-32). The radioactive photoaffinity probe was carried out using [125I-Tyr1,2,4-NAPS-Trp27]-beta h-EP and IMR-32 cell membranes. After solubilization with sodium dodecyl sulfate (SDS) and SDS polyacrylamide gel electrophoresis, a single labelled protein band was identified with a molecular weight of 72,000. Labelling was blocked by beta h-EP or beta h-EP-(6-31) but remained in the presence of beta h-EP-(1-27). The specificity of this band is thus identical to that of the non-opioid site previously characterized. Various nonionic or zwitterionic detergents did not extract the labelled non-opioid site. PMID- 2988534 TI - 2'-Nor-cGMP: a seco-cyclic nucleotide with powerful anti-DNA-viral activity. AB - As part of our study of antiherpetic acyclonucleosides, we synthesized a cyclic GMP analog, 9-[(2-hydroxy-1,3,2-dioxaphosphorinan-5-yl)oxymethyl]guanine P-oxide, sodium salt (2'-nor-cGMP), and discovered its potent and broad spectrum anti-DNA viral activities. 2'-Nor-cGMP inhibits the replication of many DNA viruses, including herpes simplex virus, human cytomegalovirus, vaccinia, SV40, and adenovirus, but does not inhibit RNA viruses. In plaque reduction studies this potent antiviral agent is also approximately 10-fold more potent than 9-(1,3 dihydroxy-2-propoxymethyl)guanine (2'NDG) against varicella-zoster virus and inhibits cell transformation by bovine papilloma virus. Unlike 2'NDG, the potent activity of 2'-nor-cGMP against herpes virus is not dependent upon the action of virus-specified thymidine kinase. Intercellular metabolism of 2'-nor-cGMP produced small amounts of 2'NDG triphosphate which were insufficient to account for the antiviral activity observed, implying that this potent anti-DNA-viral agent operates by a mechanism different from that of known acyclonucleosides. PMID- 2988535 TI - The interaction of vasoactive intestinal peptide (VIP) with isolated bovine thyroid plasma membranes. AB - The binding of vasoactive intestinal peptide (VIP) and stimulation of adenylate cyclase were studied in bovine thyroid plasma membranes. The binding depended on time, temperature and was saturable and specific. Binding studies suggested the presence of two classes of binding sites: a class with high affinity (Kd = 13 nM) and low capacity (6411 sites/pg), and a class with low affinity (Kd = 480 nm) and high capacity (105,300 sites/pg) at 15 degrees C. Secretin, glucagon, insulin and somatostatin did not displace the tracer from the membranes. VIP stimulated cyclic AMP production. Maximal cyclic AMP production (2-fold above basal values) was observed with 100 nM VIP and half-maximal response was obtained at 5 nM VIP at 15 degrees C. PMID- 2988536 TI - Desensitization of opioid-stimulated GTPase in neuroblastoma x glioma hybrid cells. AB - NG108-15 cells were pretreated with the opioid peptide [D-Ala2, D-Leu5]enkephalin and the opioid-dependent low Km GTPase was assayed in membranes. Pretreatment resulted in a small decrease in basal GTPase activity and led to a concentration dependent reduction in opioid-mediated stimulation of the enzyme. These effects were observed whether the agonist was present or absent throughout all the experimental procedures, but, in the second condition, the desensitization was smaller. The addition of naloxone had no effect on basal GTPase activity, in either control or pretreated cell membranes. Both Na+ and Mg++ were required for the opioid-induced stimulation of the GTPase. Mg++ enhanced basal enzymatic activity in controls, whereas in membranes from pretreated cells, it produced an inhibition. Thus, desensitization of the opioid-dependent low Km GTPase occurs upon chronic opioid treatment and a Mg++ regulatory site might be altered in the course of this process. PMID- 2988537 TI - Polymyxin B causes coordinate inhibition of phorbol ester-induced C-kinase activity and proliferation of B lymphocytes. AB - Lymphocytes were found to be rich in phospholipid/Ca2+-dependent (C-kinase) activity. Addition of polymyxin B (PMB) to in vitro assays of endogenous and exogenous phosphorylation resulted in profound inhibition of C-kinase activity. The phorbol ester 12-o-tetradecanoyl phorbol-13-acetate (TPA) directly activated C-kinase, leading to increased phosphorylation of the same substrates. TPA also stimulated proliferation of B cells as assessed by 3H-thymidine uptake, and PMB strongly inhibited this effect. This coordinate inhibition of TPA-induced phosphorylation and mitogenesis indicates that PMB is a potentially useful inhibitor of C-kinase activity, and that this enzyme may play an important role in mediating B cell responses. PMID- 2988538 TI - Regions of complementarity between the messenger RNAs for epidermal growth factor, transferrin, interleukin-2 and their respective receptors. AB - Messenger (m)RNA sequences complementary to the mRNA sequences for the receptors to epidermal growth factor (EGF), interleukin-2 (IL-2) and transferrin (TF) were written out and compared for homologies with their ligands (EGF, IL-2 and TF, respectively). Highly significant amino acid and nucleotide homologies between the ligands and their appropriate receptor complements were detected in each case. For example, EGF and its receptor complement contained two homologous segments, each being six amino acids in length. When these segments were screened for matches against a protein sequence bank (3060 proteins and 616,748 test segments), only EGF contained either sequence. Similar results were obtained with IL-2 and TF. In each case, the homologous segments corresponded to complementary regions in the ligand binding portion of the receptor. PMID- 2988539 TI - Effect of divalent metal ions and glycerol on the GTPase activity of H-ras proteins. AB - The product of the protooncogenic ras gene (p21N ras) exhibits a weak GTPase activity. A significant increase in the GTPase activity associated with p21N ras protein was obtained by using glycerol in the assay mixture. Of the several metal ions tested, only Mg++ and Mn++ are effective divalent cations that support the GTPase activity of p21N ras protein. p21N ras protein exhibits higher GTPase activity and yields higher [3H] GDP binding in the presence of MnCl2 than with MgCl2. Optimal GTPase and [3H] GDP binding are obtained at micromolar concentrations of MgCl2 or MnCl2. Concentrations in the millimolar range of either MgCl2 or MnCl2 are inhibitory to the GTPase activity, whereas [3H] GDP binding was not affected. PMID- 2988540 TI - Regulation by heme of the synthesis of cytochrome C oxidase subunits V and VII in yeast. AB - The effects of heme on the synthesis of subunits V and VII of cytochrome c oxidase were examined in the heme-deficient yeast mutant, GL1. In vitro translation and immunodetection with subunit-specific antisera indicated a 50% decrease in both proteins, with RNA obtained from hemeless cells. Unsupplemented, pulse-labeled cells contained both V and VII polypeptides, but at extremely low levels as compared with those found in delta-aminolevulinic acid-supplemented cells. The data suggest that heme controls the formation of mRNAs for the two subunits, and may also have a regulatory role in translation and in the stability of the polypeptides. PMID- 2988541 TI - Right-side-out pumping Na,K-ATPase-liposomes: a new tool to study the enzyme's receptor function. AB - The technology to prepare right-side-out pumping Na,K-ATPase-liposomes is described. The 50% right-side-out oriented pumps of ATP-containing liposomes are then activated by the addition of external Rb ions, leading to a ouabain sensitive Rb-influx which is the mirror-image of the inside-out transport. The resulting internal Rb concentration is 4 to 10 fold larger than the external concentration. Finally, the accumulated Rb ions can be extruded by driving the 50% inside-out oriented pumps by external ATP. PMID- 2988542 TI - EPR signals of redox active copper in EDTA washed membranes of the cyanobacterium Synechococcus 6311. AB - A signal of Cu2+ (g = 2.03) was detected by electron paramagmetic resonance spectroscopy in oxidized membrane preparations of Synechococcus 6311. The membranes were prepared and washed in the presence of EDTA (10mM, pH 8.0) and, hence, were depleted of adventitious copper; the treatment also would remove any membrane-associated soluble redox proteins and other paramagnetic metal ions. 0.1% Triton X-100 facilitated detection of the Cu2+ signal which was fully reduced by dithionite or ascorbate plus N,N,N',N',-tetramethyl-p phenylenediamine, and partially reduced NADPH and NADH, which are known to donate electrons to the terminal oxidase of cyanobacteria via the respiratory chain. Using temperature dependence and power saturation of the EPR copper signal, we conclude that copper is a firmly bound constituent of the terminal oxidase in an environment which is very similar if not identical to other cytochrome c oxidase preparations. PMID- 2988543 TI - Folate and cAMP modulate GTP binding to isolated membranes of Dictyostelium discoideum. Functional coupling between cell surface receptors and G-proteins. AB - In membrane preparations from D. discoideum cells GTP-binding activity is observed. The lack of GTP binding to intact cells suggests that the binding sites are localized inside the cell. The GTP-binding activity also remains in the particulate fraction in the presence of 1 mM Ca++. This excludes membrane associated microtubuli to be responsible for the observed GTP binding. Scatchard analysis suggests the existence of one type of binding site (Kd = 2.6 microM and 3.6 X 10(5) sites per cell). The kinetics of association as well as dissociation, however, suggest that GTP binding is more complex than binding to a single type of site. GDP and guanylyl imidodiphosphate are potent competitors of GTP binding (respectively 5- and 10-fold worse than GTP) while GMP, cGMP and several adenine nucleotides are ineffective up to 1 mM. The chemoattractants cAMP and folic acid both increase the equilibrium binding level of GTP, while dissociation of GTP is accelerated. These data indicate the functional coupling between cell surface receptors and G-proteins. PMID- 2988544 TI - Regulation of cyclic GMP levels by neurotensin in neuroblastoma clone N1E115. AB - The binding of 125I-labeled [monoiodo-Tyr3]neurotensin to intact neuroblastoma N1E115 cells and the effect of neurotensin on the intracellular concentration of cyclic nucleotides were studied at 37 degrees C and under physiological conditions of pH and ionic strength. The radiolabeled neurotensin analogue bound specifically to differentiated cells with a dissociation constant of 0.75 nM and a maximal binding capacity of 45 fmol/10(6) cells. Incubation of neuroblastoma cells with neurotensin in the presence of calcium ions resulted in a transient increase of 10 fold over basal level of the intracellular cyclic GMP concentration. Half-maximal stimulation was obtained with 2 nM neurotensin. Under identical conditions the cyclic AMP concentration only decreased by 20-30%. These results suggest that cyclic GMP is a second messenger of neurotensin in neuroblastoma clone N1E115. PMID- 2988545 TI - Methylation and expression of bovine leukemia proviral DNA. AB - In vivo, the BLV proviral DNA usually resides in a transcriptionally inert state, and is hypermethylated. Upon short-term in vitro cultivation of the neoplastic or non-neoplastic lymphoid cells, the viral genome becomes transcriptionally active but without detectable change in its methylation state. Proviral DNA was found to be methylated in one but not in the other long-term BLV producer cell line examined. These data indicate that hypermethylation of proviral DNA may not be responsible for the covert nature of BLV infection in vivo. PMID- 2988546 TI - Effect of amiodarone on membrane fluidity and Na+/K+ ATPase activity in rat-brain synaptic membranes. AB - In rat-brain synaptic membranes at a fixed temperature (37 degrees C), amiodarone dose-dependently inhibits the Na+/K+ ATPase activity (IC50 approximately equal to 2.10(-5)M) and produces a linear increase in the degree of fluorescence depolarization (P) of 1,6-diphenylhexatriene embedded in the lipid matrix. Amiodarone has no effect on Mg++ ATPase and K+PNPase activity up to 3.10(-4)M. Studies carried out at different temperatures indicate that 10(-5)M amiodarone inhibits the Na+/K+ ATPase and decreases the lipid fluidity at all the temperatures studied (9 - 40 degrees C). The compound significantly displaces the temperature of transition observed around 20 degrees C in both Na+/K+ ATPase activity and lipid fluidity to 24 degrees C with no changes in slopes. The results suggest that part of the selective inhibition of Na+/K+ ATPase activity by amiodarone could be due to the effects of the drug on lipid dynamics. PMID- 2988547 TI - Self-regulation of calmodulin-dependent protein kinase II and glycogen synthase kinase by autophosphorylation. AB - Calmodulin-dependent protein kinase II from rat brain underwent autophosphorylation and the autophosphorylation caused a marked decrease in the enzyme activity. Calmodulin-dependent glycogen synthase kinase from rabbit skeletal muscle was also inactivated by incubation under autophosphorylating conditions. The inactivation of the protein kinases by the autophosphorylation may be an important self-regulatory mechanism in controlling the enzyme activities. PMID- 2988549 TI - Lowered level of translatable messenger RNAs for manganese superoxide dismutase in human fibroblasts transformed by SV 40. AB - The activity of manganese superoxide dismutase (MnSOD) revealed by specific staining after gel electrophoresis of cell extracts, is decreased in human fibroblasts transformed by SV40. The decrease in enzyme activity is attributable to decreased amount of enzyme protein as determined by radial immunodiffusion. Total fibroblast RNAs were translated in the presence of (35S) methionine in a cell-free translation system and the neo synthesized proteins submitted to immunoprecipitation with an anti MnSOD antiserum. Gel electrophoresis of the immunoprecipitated material followed by fluorography shows that MnSOD is translated as a peptide which is 2000 daltons larger than the mature enzyme subunit. This precursor (pre-MnSOD) is processed in vitro to mature MnSOD by the action of an isolated mitochondrial preparation. Levels of translatable MnSOD mRNA in normal and SV 40 transformed cells were compared in terms of the radioactivities incorporated into pre MnSOD bands. The results indicate that the decreased amount of MnSOD in SV 40 transformed fibroblasts is due to a decreased level of translatable mRNA for MnSOD. PMID- 2988548 TI - Enzymatic synthesis and hydrolysis of [32P]phosphatidylinositol phosphate. AB - Phosphatidylinositol kinase activity in plasma membrane preparations of mouse liver was found to be comparable to that in A431 cells and higher than that in three human tumor xenografts. This activity was exploited in preparing 32P labeled phosphatidylinositol phosphate of high specific radioactivity in which approximately 4% of the radioactivity of the substrate, [gamma-32P]ATP, was incorporated into the lipid. The subcellular distribution of phosphatidylinositol phosphate phosphatase in a human astrocytoma xenograft was determined using [32P]phosphatidylinositol phosphate as a substrate. The highest phosphatase activity was found in the plasma membranes. PMID- 2988550 TI - Antiviral potency of adenosine analogues: correlation with inhibition of S adenosylhomocysteine hydrolase. AB - For a series of adenosine analogues a close correlation (r = 0.986) was found between their antiviral potency (against vesicular stomatitis virus) and their inhibitory effects (Ki/Km) on S-adenosylhomocysteine (AdoHcy) hydrolase; thus, in order of increasing inhibitory potency for both virus replication and AdoHcy hydrolase activity: (S)-9-(2,3-dihydroxypropyl)adenine less than (RS)-3-adenin-9 yl-2-hydroxypropanoic acid (isobutyl ester) less than carbocyclic 3 deazaadenosine less than neplanocin A. Our findings point to AdoHcy hydrolase as the target for the broad-spectrum antiviral activity of these adenosine analogues. PMID- 2988551 TI - Demonstration of receptors for a PDGF-like mitogen on human osteosarcoma cells. AB - U-2 OS human osteosarcoma cells synthesize, process and secrete a platelet derived growth factor (PDGF)-like mitogen. Incubation of these cells with 1 mM suramin unmasks PDGF receptor sites which are normally occupied or down regulated by the secreted endogenous PDGF-like mitogen. Partially purified preparations of metabolically labelled U-2 OS conditioned medium binds to U-2 OS cells and binding is inhibited by excess PDGF. These findings suggest that U-2 OS cells are capable of autocrine stimulation. PMID- 2988552 TI - SCH 23390 may alter dopamine-mediated motor behaviour via striatal D-1 receptors. AB - SCH 23390 potently displaced the specific binding of 3H-piflutixol to D-1 sites in striatal membranes but haloperidol was only weakly effective. SCH 23390 weakly displaced specific 3H-spiperone binding to D-2 sites, but haloperidol was potent. SCH 23390 was more effective than haloperidol in inhibiting dopamine stimulated striatal adenylate cyclase activity. These results confirm the D-1 selectivity of SCH 23390. However, SCH 23390 inhibited apomorphine-induced stereotypy and climbing behaviour in rats with equal potency to haloperidol. Haloperidol dose dependently increased striatal HVA and DOPAC concentrations without altering dopamine content. Low doses of SCH 23390 elevated striatal DOPAC concentrations but higher doses were without effect; striatal dopamine and HVA overall was unaffected by administration of SCH 23390. Haloperidol did not affect basal 3H acetylcholine release from striatal slices but reversed the apomorphine-induced inhibition of 3H-acetylcholine release. SCH 23390 did not affect basal 3H acetylcholine release nor did it reverse the apomorphine-induced inhibition of 3H acetylcholine release. The ability of SCH 23390 to inhibit motor behaviour in the rat may be due to its action on D-1 receptors since the drug does not cause typical changes in parameters of striatal D-2 receptor function. PMID- 2988553 TI - Forskolin activation of serotonin-stimulated adenylate cyclase in the liver fluke Fasciola hepatica. AB - Properties of forskolin activation of adenylate cyclase in the liver fluke Fasciola hepatica are described. Forskolin stimulated adenylate cyclase activity in cell-free fluke particles to levels more than 30-fold above the basal rate. This activation was not dependent on guanine nucleotides and, upon washing of the particles, was rapidly reversed. Forskolin potentiated the activation of adenylate cyclase by serotonin (5-HT) and lysergic acid diethylamide (LSD), resulting in both an increase in the maximal level of enzyme activity and a decrease in the apparent activation constant (KA). The 5-HT antagonist 2-bromo LSD did not inhibit enzyme activation by forskolin. Furthermore, forskolin had no effect on specific [3H]LSD binding to fluke particles. Activation of adenylate cyclase by sodium fluoride or guanine nucleotides was modified in a complex manner by forskolin with both stimulatory and inhibitory effects present. The results suggest that forskolin does not interact directly with the 5-HT receptor coupled to adenylate cyclase. Instead, it appears that forskolin effects are, at least in part, due to its ability to alter the interaction between the regulatory and catalytic components of adenylate cyclase. Incubation of intact flukes with forskolin increased their cAMP levels 2- to 3-fold. The concentration dependence of this response was similar to that for forskolin activation of adenylate cyclase in fluke particles, with 300 microM forskolin giving the maximum response. Forskolin and other agents that increased fluke cAMP levels also stimulated fluke motility. PMID- 2988554 TI - Interaction of DDT and pyrethroids with calmodulin and its significance in the expression of enzyme activities of phosphodiesterase. AB - To understand the significance of the inhibitory action of 1,1,1-trichloro-2,2 bis(p-chlorophenyl)ethane (DDT) and pyrethroid insecticides on calmodulin, a universal Ca2+ binding protein, a bovine heart phosphodiesterase-calmodulin system was studied. It was found that, at concentrations of less than 10(-5) M, the inhibitory action of DDT of the phosphodiesterase was due entirely to its action on calmodulin alone. Cypermethrin was less potent than DDT, but it also affected only calmodulin. Permethrin was the most potent inhibitor affecting calmodulin and, to a lesser extent, phosphodiesterase. The inhibitory action of these insecticides on calmodulin raises a possibility that many unsuspected Ca2+ related systems are being affected by these insecticidal chemicals, as calmodulin is known to play vital roles in many biological reactions dependent upon Ca2+. These include modulation of phosphodiesterase, neurotransmitter release, adenylate cyclase, Ca2+-dependent protein kinase, myosine light chain kinase and various membrane phosphorylation systems. PMID- 2988555 TI - Ellipticine derivatives interact with muscarinic receptors. AB - Ellipticine derivatives or analogues, tetracyclic alkaloids used in human cancer treatment, have been evaluated with regard to their interaction with several neurotransmitter receptors, in order to explain or to predict the side effects which occur in man. These drugs were recently found to be reversible non competitive inhibitors of cholinesterases. In this study, we have shown that ellipticines are also potent muscarinic antagonists, only 100-fold less active than atropine in inhibiting 50% of the specific binding of (3H) quinuclidinyl benzilate on rat brain preparation of muscarinic receptors. That the interaction with muscarinic receptors is quite unique has been demonstrated by the lack of interaction with three other neurotransmitter receptors. Tertiary amines show relatively less blockade of muscarinic receptors, while substituted ammonium ions are better inhibitors of the QNB binding. The possible mechanisms of in vivo action of these alkaloids is discussed. PMID- 2988557 TI - Enzymatic synthesis of sarin and soman. PMID- 2988556 TI - Novel microsomal anion-sensitive Mg2+-ATPase activity in rat brain. AB - Ethacrynic acid (EA) highly sensitive Mg2+-ATPase activity was demonstrated in rat brain microsomes. Marker enzyme studies suggested that the EA highly sensitive Mg2+-ATPase activity originated mainly from plasma membranes, and possibly from synaptic vesicles. Oligomycin did not affect the EA highly sensitive Mg2+-ATPase activity. Sulfhydryl reagents, such as N-ethylmaleimide and 5,5'-dithiobis-(2-nitrobenzoic acid), and anion transport inhibitors, such as 4 acetamide-4'-isothiocyanostilbene-2,2'-disulfonic acid, 4,4'-diisothiocyano stilbene-2,2'-disulfonic acid and 2,4-dinitro-1-fluorobenzene, completely inhibited the EA highly sensitive Mg2+-ATPase activity with apparent Ki values at 5, 5, 8, 8 and 10 microM respectively. Treatment of microsomes with ethylenediaminetetraacetic acid and ammonium sulfate increased the EA highly sensitive Mg2+ and Na+,K+-ATPase activities, but not EA less sensitive Mg2+- or HCO3-ATPase activity, 2- to 3-fold that in crude microsomes. Relative substrate specificities of ATP much greater than GTP greater than ITP greater than UTP, CTP, a Km for ATP at 0.77 mM, and an optimal pH at pH 7.4 were observed. Among the anions tested (Cl-, Br-, F-, HCO3-, I-, SCN-, NO3-), EA highly sensitive Mg2+ ATPase activity was stimulated significantly by Cl- and reduced by NO3-. These data suggest that a novel, plasma membrane-located and anion-sensitive Mg2+ ATPase activity exists in the brain. PMID- 2988558 TI - Mechanisms of endothelium-dependent vascular smooth muscle relaxation. PMID- 2988559 TI - Correlation between inhibition of a cyclic GMP phosphodiesterase and relaxation of canine tracheal smooth muscle. AB - Inhibition of partially purified cyclic nucleotide phosphodiesterase activity as well as pharmacologically induced relaxation of respiratory airways smooth muscle was examined to determine whether any correlation between these two effects could be found. The phosphodiesterase in extracts of canine tracheal smooth muscle was chromatographed on a DEAE Bio-Gel A column and eluted with a sodium chloride gradient. The peak I activity hydrolyzed cGMP at a higher rate than cAMP although the apparent Km values for these two cyclic nucleotides were relatively close. Comparison of the Ki values for alkylxanthine inhibitors of peak I activity correlated remarkably well with the EC50 values of the same compounds as relaxants of canine tracheal smooth muscle strips. It is concluded that inhibition of the peak I enzyme may cause accumulation of an intracellular pool of cyclic nucleotide and thus produce or contribute to the muscle relaxant effects that were observed. PMID- 2988560 TI - Inhibition of leukotriene B4 formation in human neutrophils after oral nafazatrom (Bay g 6575). AB - Three grams of nafazatrom (Bay g 6575), given orally to healthy male volunteers in a single dose, significantly reduce the formation of leukotriene B4 in polymorphonuclear leukocytes. LTB4 synthesis fell from 57.1 +/- 17.0 ng/10(7) PMNL, mean +/- S.D., in control to 34.3 +/- 14.4 ng/10(7) PMNL 3 hr after nafazatrom (2 P less than 0.001). In vitro, nafazatrom inhibited LTB4 formation in human PMNL in a dose dependent manner. At 1 microM nafazatrom LTB4 formation was reduced to 65% of the control value. Nafazatrom had no effect on the excretion of 2,3-dinor-6-keto-PGF1 alpha and 2,3-dinor-TXB2, the major urinary metabolites of endogenously synthesized PGI2 and TXA2, respectively. Serum levels of TXB2 in clotted whole blood also remained unchanged. The inhibitory effect of nafazatrom on leukotriene biosynthesis in human PMNL suggests a therapeutic potential of this drug in processes like allergy and chronic inflammation, where leukotrienes play a pathogenetic role. PMID- 2988561 TI - Obligatory role of cytochrome b5 in the microsomal metabolism of methoxyflurane. AB - Cytochrome b5 has recently been shown to be required in the reconstituted cytochrome P-450 system for the metabolism of the volatile anesthetic methoxyflurane [E. Canova-Davis and L. A. Waskell, J. biol. Chem. 259, 2541 (1984)]. To determine whether this observation in the reconstituted system was merely dependent on the particular ratios of the various components or some other fortuitous, unknown factor, or whether cytochrome b5 plays a role in the liver microsomal metabolism of methoxyflurane, the following studies were undertaken. Antibody to rabbit holocytochrome b5 was raised in guinea pigs. The antibody to cytochrome b5 was able to inhibit 75% of the microsomal metabolism of methoxyflurane. This same antibody also inhibited methoxyflurane metabolism in the reconstituted system. When the antibody to cytochrome b5 was treated with purified cytochrome b5 before addition to the microsomes, it did not inhibit methoxyflurane metabolism. Furthermore, the antibody to cytochrome b5 did not inhibit the microsomal metabolism of benzphetamine. This suggests that cytochrome b5 was required for the microsomal metabolism of methoxyflurane. It is possible that cytochrome b5 functioned in the metabolism of methoxyflurane by retaining a specific conformation of cytochrome P-450 and not by transferring the second electron to cytochrome P-450. To explore this possibility, cytochrome b5 was reconstituted with Mn3+-protoporphyrin IX. The Mn3+-protoporphyrin IX derivative retained the conformation of cytochrome b5 but not its electron transfer properties. This manganese derivative of cytochrome b5 was unable to stimulate the metabolism of methoxyflurane. The study demonstrated that cytochrome b5 was obligatory for the microsomal metabolism of methoxyflurane, whereas it was not required for the microsomal N-demethylation of benzphetamine. Moreover, the heme moiety of cytochrome b5 functioned to transfer electrons in this reaction. PMID- 2988562 TI - Effect of positively charged local anesthetics on a membrane-bound phosphatase in Acholeplasma laidlawii. AB - The plasma membrane p-nitrophenylphosphatase activity of Acholeplasma laidlawii was stimulated by the spin-labeled local anesthetic 2-[N-methyl-N-(2,2,6,6 tetramethylpiperidinooxyl)]ethyl p-hexyloxybenzoate, abbreviated as C6SL, and its methylated quaternary analog, C6SLMeI. The tertiary amine C6SL (at a concentration of 5 X 10(-5) M) was more potent at pH 6.5 than at pH 7.7. In contrast, the permanently-charged C6SLMeI was equally potent, independently of pH. These results suggest that cationic forms of the anesthetics are responsible for stimulating the enzyme. Electron spin resonance studies of C6SL- and C6SLMeI labeled membranes showed that these anesthetics in their cationic forms interacted electrostatically with components of the Acholeplasma membrane. For C6SL, this interaction was pH dependent and correlated with the pH dependency of the anesthetic-induced enzyme stimulation in the Acholeplasma membranes. Further, studies using 5-doxylstearic acid labels and non-spin-labeled anesthetics at various pH values showed that the membrane-fluidizing effect of anesthetics was not correlated with anesthetic-induced pNPPase stimulation. Our observations are consistent with the hypothesis that electrostatic interactions between cationic local anesthetics and anionic membrane components may lead to functional changes mediated by membrane proteins. PMID- 2988563 TI - Role of enhanced inositol phospholipid metabolism in neutrophil activation. AB - When guinea pig neutrophils were stimulated with chemotactic peptide [formylmethionyl-leucyl-phenylalanine (fMLP)], a marked release of lysosomal enzyme and production of superoxide anion were detected. The breakdown of phosphatidylinositol 4,5-bisphosphate (TPI) and the subsequent formation of diacylglycerol, phosphatidic acid and free arachidonic acid also occurred during the processes. Ca2+ ionophore A23187 caused an evident secretion of lysosomal enzyme but no superoxide anion production. Ca2+ ionophore also caused TPI breakdown to diacylglycerol although this breakdown was not as significant as that detected by fMLP. The tumor promotor tetradecanoylphorbol acetate (TPA), which is a strong activator of superoxide anion production but not a good stimulator of lysosomal enzyme secretion, did not cause a significant decrease of TPI or arachidonic acid release. Since TPA is known not to increase the intracellular Ca2+ level, these results suggest that lysosomal enzyme secretion is correlated closely with enhanced inositol phospholipid metabolism and Ca2+ dependent processes. On the other hand, superoxide anion production seemed to be caused mainly by Ca2+-independent processes, perhaps by protein kinase-C activation through newly formed diacylglycerol, when neutrophils were activated by chemotactic peptide. PMID- 2988564 TI - Beta-adrenergic receptors in rat mammary gland. PMID- 2988566 TI - On the site of action of the anti-adrenal steroidogenic effect of cyproterone acetate. AB - Cyproterone acetate (CA) inhibited 1-24 ACTH (50 ng/l)-stimulated cortisol production by dispersed guinea-pig adrenal cells in a dose-related manner. Inhibition occurred over the range 10(-6) to 5 X 10(-5) moles/l. The concentration of drug which induced 50% inhibition was 4.6 X 10(-6) moles/l. The sites of action of this anti-steroidogenic effect have been established. Dispersed adrenal cells were challenged with the cortisol precursor steroids (all at 10(-5) moles/l) pregnenolone (Pe), 17 alpha hydroxypregnenolone (17-OH Pe), 17 alpha-hydroxyprogesterone (17-Po), and 11 deoxycortisol or 1-24 ACTH (100 ng/1) in the absence and presence of increasing concentrations of CA (10(-5) to 10(-4) moles/l). In the absence of drug, the steroid precursors or ACTH provoked cortisol secretion greater than 10-fold that secreted by cells incubated in their absence. ACTH-stimulated cortisol secretion was inhibited greater than 68% at concentrations of CA greater than 10(-5) moles/l. CA (10(-5) moles/l) had no significant effect on steroid-stimulated cortisol production when the delta 4, 3 ketosteroids 17-OH Po and 11-deoxycortisol were used, but depressed secretion by greater than 61% (P less than 0.001) when the delta 5 3 beta hydroxysteroids (Pe, 17-OH Pe) were employed. Increasing CA concentrations to 10(-4) moles/l had little effect on cortisol secretion provoked by 11-deoxycortisol, but significantly (P less than 0.05) depressed cortisol secretion stimulated by 17-OH Po. These results suggest that the major site of action of CA is delta 5, 3 beta hydroxysteroid dehydrogenase (3 beta-HSD) with a secondary effect on 21 hydroxylase activity. To confirm these findings cortisol secretion provoked by 17, 21, dihydroxypregnenolone (17, 21 diOH Pe) and 21 deoxycortisol (21-DOC) was measured in the absence and presence of increasing concentrations of CA. At the lowest concentration of CA (5 X 10(-6) moles/l), cortisol secretion provoked by 17, 21 diOH Pe was inhibited by 28% (P less than 0.01) whereas secretion provoked by 21-DOC was not significantly affected. At the highest concentration of CA (10( 4) moles/l), the relative inhibition was 80% for 17, 21 diOH Pe and 38% for 21 DOC. We conclude that cyproterone acetate inhibits adrenal steroidogenesis at both 3 beta-HSD and 21-hydroxylase, the degree of inhibition being more pronounced at 3 beta-HSD. PMID- 2988565 TI - Reduced cyclic-AMP responsiveness in the colliculus inferior of audiogenic seizure-prone rats. PMID- 2988567 TI - Inhibition of human platelet aggregation by dihydropyrano- and dihydrofuranocoumarins, a new class of cAMP-phosphodiesterase inhibitors. AB - Certain esters of dihydropyranocoumarin and dihydrofuranocoumarin alcohols have previously been shown to inhibit the cAMP-phosphodiesterase from bovine heart. We now report that these naturally occurring coumarins inhibit the high affinity (Km = 1.1 microM) cAMP-phosphodiesterase from human platelets with activities that closely correlate with those obtained using phosphodiesterase from bovine heart tissue. Additionally the coumarins inhibit the aggregation of human platelets induced with ADP, adrenaline and collagen with activities comparable to those of dipyridamole. A lack of significant correlation between these metabolic and functional activities indicates that there exist, besides cAMP-phosphodiesterase inhibition, additional mechanisms of action for the platelet aggregation inhibitory effect of dihydropyrano- and dihydrofuranocoumarins. PMID- 2988569 TI - [High performance liquid chromatography of peptide bioregulators, their fragments and derivatives. II. Sorption of peptides from aqueous-organic eluents on octadecylsilylsilica gel and unmodified silica gel]. AB - Dependence of the peptide retention upon the organic component concentration in eluent has been studied. A parabolic dependence has been found in a wide range of acetonitrile concentrations. The effect observed with ODS- and unmodified silica as stationary phases extends analytical and preparative potentialities of HPLC of peptides. PMID- 2988568 TI - Cystine depletion by WR-1065 in cystinotic cells. Mechanism of action. AB - Cystinotic leucocytes and skin fibroblasts incubated with the aminothiol N-(2' mercaptoethyl)-1,3-propanediamine (WR-1065) exhibited substantial intralysosomal cystine depletion within 2 hr. Wr-2721, the thiol phosphorylated derivative of WR 1065, did not lower cystinotic leucocyte cystine in 1 hr but depleted cystinotic fibroblasts of cystine after 21 hr. Concentrations of cysteamine (beta mercaptoethylamine) equimolar with those of WR-1065 depleted cystine more rapidly than did WR-1065, but the extent of cystine depletion by WR-1065 approached that for cysteamine when longer periods of incubation or higher concentrations were used. Cystine depletion by WR-1065 was slower for leucocyte lysosomal granular fractions than for whole leucocytes. L-[35S]Cystine-labeled fibroblasts exposed to WR-1065 exhibited new compounds not seen when cells were incubated without WR 1065: WR-1065-cysteine, cysteamine-cysteine and cysteamine-glutathione mixed disulfides. L-[35S]Cystine-loaded lysosome-rich granular fractions from cystinotic leucocytes incubated with WR-1065 formed WR-1065-cysteine mixed disulfide but no cysteamine-cysteine mixed disulfide. We suggest that WR-2721 is dephosphorylated intracellularly to the free thiol, WR-1065, which subsequently is converted to cysteamine by an unknown route. Intracellular cysteamine then enters the lysosome and reacts with free cystine to form cysteamine-cysteine mixed disulfide and cysteine which move into the cytosol and the incubation medium where they participate in further interchange reactions with free thiols present there, namely WR-1065 and glutathione. PMID- 2988570 TI - [A plasmid vector with temperature-controlled gene expression]. AB - A 169 b.p. fragment including the bla gene promoter p3 has been removed from pBR327 plasmid, and the deleted plasmid used for cloning the TaqI/BglII-fragment of the lambda c1857ind- DNA containing promoter pR and gene cI to obtain plasmid pCE119. Cells containing pCE119 produced a high level of beta-lactamase at 42 degrees C, the yield at 42 degrees C being 100 times higher than at 32 degrees C. For cloning and functional assays a pCEZ12 plasmid was constructed, in which promoter pR and repressor cI of lambda phage control the expression of the semi synthetic beta-galactosidase gene. Yield of beta-galactosidase produced by pCEZ12 at 42 degrees C was ca. 300 times higher than at 32 degrees C. PMID- 2988571 TI - Antibodies to human T cell leukemia virus are absent in patients with systemic lupus erythematosus. AB - It has been anticipated that type C oncornaviruses, which participate in the pathogenesis of murine systemic lupus erythematosus (SLE), would be found to have a role in the development of SLE in humans. In studies of tissues from SLE patients, type C-related proteins have been identified. Using information obtained on the recent isolation of the human T cell leukemia virus (HTLV), together with that from studies of murine SLE, we attempted to clarify the role of antibodies to HTLV in the pathogenesis of human SLE. Using a solid-phase enzyme-linked immunosorbent assay and sodium dodecyl sulfate-polyacrylamide gel electrophoretic techniques, we were unable to find evidence of the participation of antibodies to HTLV proteins in the development of human SLE. PMID- 2988572 TI - Synovial fluid crystals in osteoarthritis. AB - Apatite crystals, calcium pyrophosphate dihydrate crystals, or both were observed in knee joint effusions from 60% of 100 consecutive osteoarthritis (OA) patients. Crystals were mor common in patients with more severe OA and in joints that had received previous intraarticular steroid injections. Whether the latter was predominantly related to an effect of the steroid or to the fact that steroids were used in more severe cases was not clear. Crystals existed in many effusions without elevated synovial fluid leukocyte counts. A subgroup of 7 patients had both types of crystals, more crystal phagocytosis, and more severe OA. PMID- 2988574 TI - Cartilage surface charge. A possible determinant in aging and osteoarthritic processes. AB - Polycationic labels such as cationized ferritin and colloidal iron were used to evaluate the surface negative charges over the mandibular condyles of ICR mice. The effects of neuraminidase, hyaluronidase, pronase, and collagenase on the binding of cationized ferritin and colloidal iron particles to the condylar articular surface were also studied. The results of this study clearly indicate that the surface area of the cartilaginous condyle is negatively charged and that its composition consists mainly of a collagenous material embedded within a proteinaceous matrix. With age, a substantial decrease in the density of negative charges took place along the surface area and, in particular, in the context of sialic acid residues. It is, therefore, possible that the reduction in cartilage surface charge might be associated with the onset of osteoarthritic changes commonly seen in aging humans and experimental animals. PMID- 2988573 TI - Role of synovial membrane inflammation in cartilage matrix breakdown in the Pond Nuki dog model of osteoarthritis. AB - Using the Pond-Nuki procedure, osteoarthritis was induced in 26 crossbred dogs. Their right knees were subjected to anterior cruciate ligament sectioning, and their left knees received a sham operation. The unoperated knees of 7 additional dogs served as controls. Cartilage and synovial membranes were excised 2, 4, and 8 weeks after surgery. Collagenolytic activity, determined by direct tissue assay, was higher at all times tested in osteoarthritic cartilage and synovia than in sham operated and control specimens. The increased collagenolytic activity of the cartilage did not correlate with the collagenolytic activity of the synovium, but it did correlate (r = 0.57) with the degree of synovial inflammation, which was graded histologically. Treatment for 4 weeks with prednisone (0.20-0.25 mg/kg/day) blocked the increased collagenolytic activity of the cartilage. Our results indicate that stimulating factors may, but collagenolytic enzymes probably to not, diffuse from the synovium to the cartilage and modulate tissue breakdown. Prednisone may also suppress cartilage breakdown by either acting at the level of the synthesis or by acting on the release/action of the stimulating factors. PMID- 2988575 TI - Calcium pyrophosphate dihydrate crystal deposition in rheumatoid arthritis. PMID- 2988576 TI - Relationship between changes in plasma lipoprotein concentrations and fecal steroid excretion in man during consumption of four experimental diets. AB - Limited information is available on the mechanism by which changes in nutrient intake influence plasma lipids. We compared the effects on plasma lipoprotein levels of 3 dietary modifications involving changes in total fat intake (27-40% of calories), cholesterol intake (100-250 mg/1000 kcal), the dietary polyunsaturated to saturated fatty acid ratio (0.3-1.0) and intake of vegetable derived fiber and protein. On these 3 diets, plasma low density lipoprotein was reduced by 26-34%. Fecal bile acid excretion was similar on all diets (363-379 mg/day). There was no alteration in fecal bile acid output associated with an increase in polyunsaturated or total fat intake. Sterol balance became significantly more negative during consumption of only 1 of the 3 cholesterol lowering diets. The observed reduction in plasma cholesterol levels was not associated with an increase in fecal bile acid output suggesting that diet induced changes in circulating cholesterol are not maintained by an increase in sterol turnover but may reflect alterations in hepatic cholesterol and lipoprotein synthesis. PMID- 2988577 TI - Trigger and amplification mechanisms in visual phototransduction. PMID- 2988578 TI - Organization of glycosphingolipids in bilayers and plasma membranes of mammalian cells. AB - The evidence presented in this review strongly suggests that, when present as a minor component in liquid crystalline phospholipid bilayers, neutral glycosphingolipids are segregated into compositional domains of small size dispersed in the matrix phospholipid. In many instances the glycosphingolipid in the dispersed domains is in the gel state. Because these domains are in the gel state, the individual molecules escape only very slowly from the surface of the bilayer, much more slowly than do the phospholipid components. There is as yet no direct evidence that this slow escape rate is a property of neutral glycolipids in biological membrane bilayers. If it is, however, then these molecules are well suited for their putative role as cell surface markers, a role that involves them in many important biological functions. There is evidence to suggest that molecules of this type are also present in a dispersed microdomain structure on the external surface of at least some mammalian cell plasma membranes. These small domains of glycolipids with their sugar residues projecting outward from the cell surface are much like a large membrane glycoprotein when viewed from the ambient medium near the cell surface. Thus, whether the sugar residues be of glycoprotein or glycolipid origin, they are localized in groups or patches on the external surface of the cell. One important consequence of this patch structure may be in the obvious effect on the free energy of binding a ligand to a patch. Whether the ligand is mono- or polyvalent, the roughly 2 M concentration of sugar in the surface patch will cause the apparent ligand binding free energy to be substantially larger than it would be for a single isolated sugar residue on the surface. In contrast to the neutral glycosphingolipids (and sulfatides, perhaps) the available information suggests that gangliosides are not localized in small domains in model systems and most probably not in biological membranes. Capping of this type of glycosphingolipid does appear to occur under certain circumstances. However, it is almost certain that capping is not an intrinsic property of ganglioside phospholipid-bilayer systems. Although at 37 degrees C gangliosides rapidly transfer from micelles to phospholipid vesicles and to cell membranes, nothing is known about the rates at which this class of molecules leave a phospholipid bilayer. Their known biological functions on the cell surface appear to require that they leave very slowly, if at all, as do the neutral glycosphingolipids. The glycosphingolipids are, by virtue of their polysaccharide moeity, a unique class of lipids and cell surface components.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2988579 TI - Retroperitoneal fibrosis and inferior vena cava obstruction: radionuclide venography (99m Tc pertechnetate). PMID- 2988580 TI - Effects of tifluadom on passive avoidance behaviour in DBA/2 mice. AB - The effects of the selective opiate kappa-receptor agonist tifluadom on memory were investigated in a passive avoidance task in 3 sets of experiments carried out with DBA/2 (DBA) mice both familiarized and unfamiliarized with the apparatus. In a first set of experiments, tifluadom (1.0 or 2.5, but not 0.5 mg/kg) administration immediately after training impaired retention performance of non-familiarized mice. This impairment was still evident when the drug was injected 15 or 30, but not 60 min after training. A second set of experiments was carried out with mice familiarized with the apparatus. Tifluadom was less effective in impairing memory in this group of animals, as compared with non familiarized mice. Finally, in a third set of experiments, carried out with non familiarized mice, a 15 min immobilization stress, which was ineffective when administered alone, enhanced the effects of tifluadom (1.0 mg/kg). The results are discussed in terms of attenuation of emotionality, resulting in impaired retention, following post-training opiate administration. PMID- 2988581 TI - Stimulation of aggression in male mice by alpha-MSH and its relation to light phase and to saline intake effects. AB - The resident/intruder test was used to examine the social approach and aggressive behaviour of male albino mice. Digging, self-grooming and rearing were also recorded, as was the post-test response to hot-plate exposure. The resident mice were given either a single acute injection of MSH (MSH); 0.9% NaCl to drink (for 48 h prior to testing; SAL); a combination of both treatments (MSH + SAL) or an injection of 0.9% physiological saline (control group; CON). Testing was carried out at the midpoints of the light and dark phases of the 12:12 light cycle. Data on plasma ion levels and hypothalamic cAMP levels were collected after the hot plate test. MSH stimulated fighting both in the light and dark, and SAL in the light. MSH + SAL reversed the effects of the single treatments in that fighting declined below CON levels in both light and dark. Social contact and other behaviours were much less affected by treatments. Pain responding and plasma ion levels were not changed. Effects on cAMP were largely inconclusive, but intruders had significantly lower levels than the aggressive resident animals. PMID- 2988583 TI - Studies on cytochrome c oxidase, XI. The amino-acid sequence of bovine heart polypeptide VIc. AB - The complete primary structure of the cytoplasmically synthesized polypeptide VIc from beef heart cytochrome c oxidase was determined via isolation and sequencing of overlapping methionine and glutamic acid fragments. The protein consists of 73 amino acids (Mr 8 480). Through the protein contains, from residues 21 to 40, a hydrophobic sequence interrupted by one lysine it may not penetrate the membrane. A sequence of 33 amino acids highly homologous to the C-terminal part of VIc has been translated from a cDNA clone of a nuclear coded subunit of the enzyme from rat liver. The function of this component of the terminal oxidase is yet unknown. PMID- 2988582 TI - Purification, amino-acid sequence and some properties of the ferredoxin isolated from Bacillus acidocaldarius. AB - Ferredoxin was isolated from the aerobic, thermophilic and acidophilic bacterium Bacillus acidocaldarius and its sequence of 78 amino acids completely determined by automated Edman degradation of the protein and of peptides derived from chemical cleavage between aspartic acid and proline and from enzymatic digestions. The optical spectrum of the oxidized protein has a broad maximum around 400 nm. The ferredoxin is thermostable: its absorbance begins to decrease only at incubation over 71 degrees C. The number of iron and inorganic sulphur atoms per molecule was determined to be 5.3 and 5.0, respectively. The calculated molar extinction coefficient was 23 000 M-1 X cm-1, the molecular mass of the apoferredoxin 8 872 Da. Contrary to all expectations, the sequence of B. acidocaldarius ferredoxin shows very little homology to that of B. stearothermophilus but closely resembles that of Thermus thermophilus. PMID- 2988584 TI - Metabolic zonation in liver of diabetic rats. Zonal distribution of phosphoenolpyruvate carboxykinase, pyruvate kinase, glucose-6-phosphatase and succinate dehydrogenase. AB - The activities and zonal distribution of key enzymes of carbohydrate metabolism were studied in livers of diabetic rats. 48 h after alloxan treatment the following alterations were observed, intermediate values being reached after 24 h: Blood glucose, acetoacetate and beta-hydroxybutyrate were increased to more than 500%; liver glycogen was reduced to about 10%. Portal vein insulin was reduced to below 10%, portal glucagon was increased to almost 200%. The glucogenic enzymes phosphoenolpyruvate carboxykinase and glucose-6-phosphatase were enhanced to 320% and 150%, respectively. The glycolytic enzymes glucokinase and pyruvate kinase L (differentiated from the M2 isoenzyme with a specific anti L-antibody) were lowered to 50% and 75%, respectively. The citrate cycle enzyme succinate dehydrogenase remained unchanged. The normal periportal to perivenous gradient of phosphoenolpyruvate carboxykinase of about 3:1, as measured in microdissected tissue samples, was enhanced to about 4:1 with activities elevated to 230% and 190%, respectively, in the two zones. The normal periportal to perivenous gradient of pyruvate kinase L of about 1:1.7, as determined with the microdissection technique, was reduced to about 1:1.4 with levels lowered to 55% and 45%, respectively, in the two zones. The even zonal distribution of pyruvate kinase M2 remained unaltered.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988585 TI - [Mechanism of ischemic disorders of fatty acid oxidation in heart mitochondria]. AB - The article deals with the effect of permanent, temporal and total myocardial ischemia on the oxidation of acetate, hexanoate, palmitoylcarnitine and palmitoyl CoA in isolated rabbit heart mitochondria. All the three models of ischemia in different experimental situations demonstrated a similar degree of fatty acid oxidation suppression independent of the length of acyl residue, the suppression being the greatest during the first hours and in total ischemia. Both in the control and in ischemia, respiratory activity was at its highest level with acetate, decreasing in the order above. Thus, the rate of Krebs' cycle reactions and of respiratory chain does not limit medium- and long-chain fatty acid oxidation. It is nevertheless established that suppression of their oxidation in ischemia is completely determined by the decrease of cytochrome S and of endogenous substrate intermediates of Krebs' cycle in the mitochondria; decreased adenine nucleotide and carnitine-palmitoyl-transferase transport (other authors' data) is not critical, at least in early ischemia (0.5 h). Ischemic mitochondria are characterized by incomplete palmitoylcarnitine oxidation. PMID- 2988586 TI - E. coli adherence to CER cells infected by vesicular stomatitis virus. AB - Research was carried out on the adherence of a mannose-resistant uropathogenic E. coli strain to CER cells infected with vesicular stomatitis virus (vsv). A decrease in the bacterial adhesion was noticed during the early phases of viral infection, probably due to a close relationship between cell receptors for VSV and E. coli, both containing glycolipids and phospholipids. In a later phase of viral infection, on the contrary, corresponding to the appearance on the cell surface of newly synthesized viral antigens, bacterial adherence was enhanced. This last observation suggested a possible predisposition of the host to bacterial colonization during renal viral infection. PMID- 2988587 TI - [Blood-brain barrier opening microcirculation in human brain tumor]. AB - The changes of the human tumor microcirculation after blood-brain barrier (BBB) opening was studied. BBB opening was performed by retrograde injection of hyperosmotic solution, such as mannitol or glycerol, from superficial temporal artery in the operation procedure. Serial fluorescein micro angiography of human tumor surface was taken by motor-driven camera equipped to surgical microscope. Two glioblastomas, two astrocytomas and one metastatic tumor were studied. In glioblastoma multiforme, after BBB opening, fluorescein sodium perfusion was increased with the enlargement of the venules and extravasation of fluorescein sodium was also observed from venules by the late venous phase of fluorescein microangiography. In low grade astrocytoma, after BBB opening, fluorescein sodium perfusion was relatively decreased in the vasculature which diameter was from 5 micron to 10 micron and no extravasation of fluorescein sodium was observed from the venules. In metastatic brain tumor, extravasation of fluorescein sodium was observed, ever before BBB opening, especially from arterial capillary or venous capillary. After BBB opening, extravasation of fluorescein sodium from those capillaries was increased and the tumor staining of fluorescein sodium was observed after BBB opening. But in low grade astrocytoma, BBB disruption was not observed. The effect of BBB disruption was more marked in malignant tumor, but little effect was observed in low grade astrocytoma and normal brain tissue. The degree of BBB disruption is depend on the volume and speed of hyperosmotic solution. Using our methods of volume and speed, we could observed the different BBB disruption according to the malignancy of the tumor. The difference of BBB reaction to hyperosmotic solution between low grade astrocytoma and malignant tumors is their stability of the BBB.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988588 TI - Nitrous oxide inhalation does not influence plasma concentrations of beta endorphin or Met-enkephalin-like immunoreactivity. AB - The possibility that nitrous oxide releases endogenous opioid peptides into the circulation has been tested in 10 pain-free, unstressed volunteers breathing 30% nitrous oxide in oxygen. Despite achieving plateau concentrations in venous blood, accompanied by subjective effects, there were no significant changes in plasma concentrations of immunoreactive beta-endorphin, methionine-enkephalin or ACTH. These results indicate that, in the absence of nociceptive input, the effects of the inhalation of nitrous oxide are unrelated to alterations in peripheral concentrations of these endogenous opioid peptides. PMID- 2988590 TI - Enalapril (MK421) activation in man: importance of liver status. AB - The in vitro conversion of enalapril (MK421) to enalaprilic acid (MK422) in human autopsy tissues was examined. MK422 was measured by radioimmunoassay. Human cadaver liver was the only tissue in which significant conversion was demonstrated. The esterase activity was stable after post mortem. Autopsy liver tissues from patients with elevated ante mortem liver function tests were found to have a reduced rate of deesterification. PMID- 2988589 TI - Effective dose range of enalapril in mild to moderate essential hypertension. AB - The dose-response relationship of enalapril was evaluated in a double-blind, balanced, two-period, incomplete-block study in 91 patients with mild to moderate essential hypertension. Patients were randomly assigned to two of six treatments: placebo, 2.5, 5, 10, 20 and 40 mg/day of enalapril maleate. There were two 3-week treatment periods, each preceded by a 4-week, single-blind placebo washout. Each dose of enalapril produced significant decreases in standing and supine systolic and diastolic blood pressure after 2 and 3 weeks of treatment. There were no significant changes on placebo. There was a significant linear dose response relationship for both mean blood pressure and mean change from baseline in blood pressure (P less than 0.01 for systolic and mean arterial pressure, and P less than 0.05 for diastolic pressure). Enalapril was associated with an increasing dose-response relationship across the 2.5-40 mg/day range. The 2.5 mg/dose is effective in some patients; however, doses greater than or equal to 10 mg/day may be necessary to achieve satisfactory blood pressure control. PMID- 2988592 TI - Production of plasminogen activators by human T-cell leukaemia virus-transformed human T cell lines. AB - Six human T cell lines HAMA, KUN, KAN, TCL-Haz, TCL-Ter, and TCL-Mor, which were transformed by a retrovirus, human T-cell leukaemia virus (HTLV), constitutively produced plasminogen activators (PAs) in culture supernatants. The amount of PAs produced varied among the cell lines. The PAs were distinguished by immunochemical analysis between two types: urokinase (UK)-type and non-UK-type. KUN, TCL-Ter, and HAMA mainly produced UK-type PA, whereas the other cell lines produced both types. Thus, HTLV-transformed T cell lines differ in the quality and quantity of the PAs they produce. The PAs in the culture supernatants of each cell line were separated into several mol. w forms on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The results indicate that the same cell line produces PAs of different mol. wt. PA production by these cell lines was affected by treatment with phorbol miristate acetate, concanavalin A, and phytohaemagglutinin; the effects were substantially different in each cell line. The data described here indicate that HTLV-transformed T cell lines constitutively produce PAs which are very heterogeneous in both quality and quantity. PMID- 2988591 TI - Pre-dosing plasma concentrations and beta-adrenoceptor blocking effects during repeated once daily dosing with 160 mg sustained-release propranolol (Inderal LA) and 16/260 oxprenolol Oros to healthy volunteers. AB - Plasma drug concentrations and attenuations in sub-maximal exercise tachycardia were measured in six healthy subjects 24 h after the third and fourth doses of Inderal LA and 16/260 oxprenolol Oros administered once daily. Both drugs significantly reduced exercise heart rates relative to the placebo response, at both observation times. The mean percentage reductions on the third and fourth days were, respectively, 15.7 and 11.6% for Inderal LA compared with 18.1 and 12.2% for 16/260 oxprenolol Oros. The differences between formulations were not statistically significant. The mean plasma concentrations at 24 h on the third and fourth day were 119 and 101 ng/g for oxprenolol and 16.2 and 15.8 ng/g for propranolol. PMID- 2988593 TI - Measurement of blood supply to murine tumours using in vivo red cell labelling and dynamic scintigraphy. AB - Blood pool and flow were studied in transplanted adenocarcinomas on the legs of mice. The animals' red blood cells were labelled in vivo by consecutive injections of a stannous compound and 99mTc-pertechnetate. The distribution of radioactivity was then recorded continuously with a gamma camera. This method allows prolonged and repeated estimations of blood supply to undisturbed tumours in conscious mice. It was found that in small tumours (under 1 ml) circulating blood pool was usually high, often 2 or 3 times that in normal leg tissues. In tumours bigger than 1 ml blood pool per unit volume tended to be lower but was still about 1.5 times the normal tissue level. This relatively large blood volume would seem to be outweighed by a very slow rate of flow. Even in the small tumours blood perfusion was greatly reduced compared to that in the normal leg. The blood pool results here provide no evidence that in tumours larger than 1 ml blood supply decreased progressively with growth. PMID- 2988594 TI - Lung tumour cell lines synthesizing peptide hormones established from tumours of four histological types: characterization of the cell lines and analysis of their peptide hormone production. AB - Thirty permanent and more than 60 primary tumour cell lines were established from pleural and pericardial exudates or wedge biopsies from human bronchial carcinoma. The permanent cell lines have their origin in 6 small cell, 5 large cell, 9 squamous and 5 adeno carcinomas of the lung. Tumour cells were purified from non tumour cells by direct cloning in fluid cultures or by soft agar cloning. In vitro secretion of ACTH, bombesin, calcitonin, and neurotensin was demonstrated for lung tumour cells belonging to the four major histological types. Cell suspensions of peptide hormone secreting permanent cell cultures were grown to solid tumours after xenotransplantation into nude mice. Comparative ultrastructural examination of the primary tumour and of cells grown in tissue culture and in xenografts demonstrated the preservation of most tumour type specific structural criteria in the ex vivo/in vitro systems. The present data show that not only tumour cells from small cell carcinoma but also from other histological types are capable of synthesizing a broad spectrum of immunoreactive peptide hormones. This result might be interpreted as indicating a common expression of hormone biosynthesis and secretion by all lung tumours. PMID- 2988595 TI - Salmon and human calcitonin like material in lung cancer. PMID- 2988596 TI - Treatment of chromomycosis with itraconazole. PMID- 2988597 TI - Persistent Epstein-Barr virus infection associated with monosomy 7 or chromosome 3 abnormality in childhood myeloproliferative disorders. AB - This report deals with myeloproliferative disorders associated with chronic, persistent Epstein-Barr virus (EBV) infection and with monosomy 7 and aberrations concerning chromosomes 3 and 5. Altogether five children were affected, their age ranging from 1 to 4 years at time of clinical diagnosis. Principal symptoms were: hepatomegaly, splenomegaly, recurring upper respiratory tract infection and anaemia. The serum IgG level remained persistently increased. Anti EBV antibody concentrations were measured over a period of 9 months to 6 years, demonstrating persistently increased concentrations of IgG antibodies to viral capsid antigen (VCA) and against early antigen (EA). In three patients IgA antibodies were also studied and were found to be elevated. Within 2-5 years two children developed chronic myelomonocytic leukaemia from the chronic myeloproliferative syndrome. A third patient who initially was diagnosed as chronic myelomonocytic leukaemia developed acute leukaemia within a period of 12 months. A fourth patient with myeloproliferative syndrome went into spontaneous remission after an observation period of 2 years. A fifth patient, the only one with translocation t(3;5)(q27;q33), displayed symptoms and a clinical course that can best be characterized as juvenile chronic myelocytic leukaemia. The clinical, haematological, serological and cytogenetic findings may be related. PMID- 2988598 TI - Ultrastructural localization of transferrin, transferrin receptor, and iron binding sites on human placental and duodenal microvilli. AB - Ultrastructural methods were used to determine the subcellular location of the transferrin receptor, transferrin and iron-binding sites on human term placenta and human duodenum microvillus surfaces. The transferrin receptor and transferrin were localized by immunocytochemical methods employing either OKT9, a human transferrin receptor monoclonal antibody, or mouse anti-human transferrin (ATfn), both followed by a horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (GAM-HRP) and diaminobenzidine (DAB) sequence. Iron-binding sites were localized by acid ferrocyanide (AF) staining after saturation of tissue specimens with iron, accomplished with iron nitrilotriacetate (FeNTA), a known transferrin iron donor. Placental microvillus surfaces demonstrated staining for the OKT9-GAM-HRP DAB-reactive transferrin receptor, ATfn-GAM-HRP-DAB-reactive transferrin, and FeNTA-AF-reactive iron acceptor, whereas enterocyte microvillus surfaces lacked significant staining with each of these methods. FeNTA-AF stained iron-binding substance in placental and enterocyte microvilli and cytoplasmic matrix. Thus using the same ultrastructural immunostaining and cytochemical methods transferrin receptor, transferrin, and nitrilotriacetate iron acceptor sites can be demonstrated on the microvillus surface of human placenta but not on the microvillus surface of human duodena. PMID- 2988599 TI - AIDS, infectivity, and health care workers. PMID- 2988600 TI - Ethylene oxide induces central-peripheral distal axonal degeneration of the lumbar primary neurones in rats. AB - Wistar rats subjected to a single exposure lasting six hours to ethylene oxide (EO) at a concentration of 500 parts per million three times a week for 13 weeks developed ataxia in the hindlegs. Myelinated fibres in hindleg nerves and in the fasciculus gracilis showed axonal degeneration sparing the nerve cell body of the lumbar dorsal root ganglion and myelinated fibres of lumbar dorsal and ventral roots. These pathological findings are compatible with central-peripheral distal axonal degeneration. This is the first animal model of EO neuropathy to be histopathologically verified. PMID- 2988602 TI - Abnormal uterine bleeding as a presenting sign of metastatic signet ring cell carcinoma originating in the breast. Case report. PMID- 2988601 TI - Plasma catecholamines and modes of delivery: the relation between catecholamine levels and in-vitro platelet aggregation and adrenoreceptor radioligand binding characteristics. AB - Catecholamines were measured in maternal venous, and mixed umbilical cord blood. Maternal catecholamines were significantly (P less than 0.01) reduced by epidural analgesia with a 36% reduction in noradrenaline and a 33% reduction in adrenaline. Fetal catecholamines were elevated at birth with a 3-8 fold increase in noradrenaline but not adrenaline during spontaneous vaginal delivery. The lowest fetal catecholamines were obtained in the group delivered under epidural analgesia; lower plasma catecholamines were not associated with adverse respiratory effects. Fetal platelets showed impaired alpha 2-adrenoceptor function with absent aggregatory responses to adrenaline in vitro. The defect in platelet function was unlikely to be related to changes in the number of fetal platelet alpha-receptors or to changes in receptor affinity for adrenaline, as fetal platelets failed to aggregate to adrenaline from deliveries with high and low cord blood catecholamines. PMID- 2988603 TI - A case report of neuroectodermal tumour of infancy. PMID- 2988604 TI - Differences in the association of calmodulin with cyclic nucleotide phosphodiesterase in relaxed and contracted arterial strips. AB - Changes in the concentration of cytosolic Ca2+ are assumed to alter the activity of Ca2+-calmodulin-sensitive cyclic nucleotide phosphodiesterase in intact cells. However, this assumption is based on indirect evidence and by analogy from studies of enzyme activities in broken cell systems. We have developed a procedure for estimating the fraction of Ca2+-calmodulin-sensitive phosphodiesterase that is in an activated, ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) sensitive state in intact porcine coronary artery strips. The experimental approach involves homogenization of the strips and assay of cyclic guanosine monophosphate (cyclic GMP) phosphodiesterase activity under conditions that retard changes in the amount of the complex Ca2+ calmodulin-phosphodiesterase. Our findings indicate that cyclic GMP phosphodiesterase in intact coronary artery strips does associate with Ca2+ calmodulin and that interventions that change the concentration of Ca2+ in the cytosol of the intact strip change the extent of this functional association. Exposure to histamine (10 or 100 microM) or 50 mM KCl caused contraction and an increase in EGTA-sensitive cyclic GMP phosphodiesterase activity. Isoproterenol induced relaxation of tissues that had been caused to contract with 10 microM histamine was accompanied by a reduction in EGTA-sensitive cyclic GMP phosphodiesterase activity to the same level as that present before contraction was initiated. PMID- 2988605 TI - Determination of the screw sense specificity of bovine liver fructokinase. AB - Fructokinase from beef liver showed a clear reversal in specificity when the two isomers of ATP beta S were used as substrates with Mg2+ and Cd2+, with the Sp isomer having the higher V/K value with Mg2+ and the Rp isomer the higher value with Cd2+. The delta isomer of MgATP is thus the active form of the substrate. The substitution of sulfur for oxygen in the noncoordinated position of the beta phosphate caused a 102-fold decrease in V/K over the value seen with MgATP, while substitution in the coordinated position gave a 21-fold decrease over the V/K value seen with CdATP. The Km values were little affected by sulfur substitution, showing that the wrong screw sense isomers were nonproductively bound almost as well as the correct ones. When ADP alpha S was used as a substrate in the reverse reaction, the Sp isomer showed the highest V/K value with both Mg2+ and Cd2+, suggesting that the metal ion is not coordinated to the alpha-phosphate during transphosphorylation. The failure of CrATP to act as a substrate for fructokinase suggests that the enzyme inserts one of its side chains into the inner coordination sphere of the metal ion during the reaction. PMID- 2988606 TI - Unambiguous stereochemical course of rabbit liver fructose bisphosphatase hydrolysis. AB - The stereochemical course of rabbit liver fructose bisphosphatase (EC 3.1.3.11) was determined by hydrolyzing the substrate analogue (Sp)-[1-18O]fructose 1 phosphorothioate 6-phosphate in H(2)17O, incorporating the chiral, inorganic phosphorothioate product into adenosine 5'-O-(2-thiotriphosphate) (ATP beta S), and analyzing the isotopic distribution of 18O in ATP beta S by 31P NMR. The result indicates that the 1-phosphoryl group is transferred with inversion of configuration. A series of single-turnover experiments ruled out an acyl phosphate intermediate in the hydrolysis. Consequently, fructose bisphosphatase catalyzes the hydrolysis of fructose 1,6-bisphosphate via a direct transfer of the phosphoryl moiety to water. PMID- 2988607 TI - Factor 390 chromophores: phosphodiester between AMP or GMP and methanogen factor 420. AB - Two chromophores with absorbance maxima at 390 nm (factors 390) have been isolated from oxidized cells of Methanobacterium thermoautotrophicum delta H. The isolation procedure included anion-exchange chromatography of the soluble cofactor pool followed by reverse-phase chromatography. The factor 390 species are novel derivatives of methanogen coenzyme factor 420 in which the 5 deazaflavin 8-hydroxy group is in a phosphodiester linkage to adenosine 5' phosphate or guanosine 5'-phosphate. The structural assignments were based, in part, on the UV-visible and 1H NMR spectra. In addition, the results from amino acid analysis, phosphate determination, 31P NMR spectroscopy, and fast atom bombardment mass spectrometry were consistent with the proposed structures. Confirmation of the factor 390 structures was made following phosphodiesterase release of the nucleotide monophosphates from factor 420. The nucleotide monophosphates were identified as AMP and GMP by UV-visible spectra and based on elution position by using reverse-phase and anion-exchange high-performance liquid chromatography. The presence of AMP was further demonstrated by using adenylate-5'-phosphate kinase which induced a spectral shift during conversion of the sample to IMP. In addition, the presence of GMP was established by a specific enzymatic assay. PMID- 2988608 TI - Protection of amide protons in folding intermediates of ribonuclease A measured by pH-pulse exchange curves. AB - pH-pulse exchange curves have been measured for samples taken during the folding of ribonuclease A. The curve gives the number of protected amide protons remaining after a 10-s pulse of exchange at pHs from 6.0 to 9.5, at 10 degrees C. Amide proton exchange is base catalyzed, and the rate of exchange increases 3000 fold between pH 6.0 and pH 9.5. The pH at which exchange occurs depends on the degree of protection against exchange provided by structure. Pulse exchange curves have been measured for samples taken at three times during folding, and these are compared to the pulse exchange curves of N, the native protein, of U, the unfolded protein in 4 M guanidinium chloride, and of IN, the native-like intermediate obtained by the prefolding method of Schmid. The results are used to determine whether folding intermediates are present that can be distinguished from N and U and to measure the average degree of protection of the protected protons in folding intermediates. The amide (peptide NH) protons of unfolded ribonuclease A were prelabeled with 3H by a previous procedure that labels only the slow-folding species. Folding was initiated at pH 4.0, 10 degrees C, where amide proton exchange is slower than the folding of the slow-folding species. Samples were taken at 0-, 10-, and 20-s folding, and their pH-pulse exchange curves were measured.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988609 TI - Recent advances in tumour imaging. Use of radiolabelled antitumour antibodies. PMID- 2988610 TI - The effect of mono- and divalent salts on the rise and decay kinetics of EPR signal II in Photosystem II preparations from spinach. AB - The rise and decay kinetics of EPR signal II have been used to probe the organization of the donor side of Photosystem II (PS II) before and after extraction of PS II preparations with high concentrations of salt. 800 mM NaCl or 500-800 mM NaBr substantially depletes the preparations of the 16 and 24 kDa proteins and decreases the steady-state rate of O2-evolution by 70-80% from control rates. These treatments do not largely alter the decay kinetics of Signal II; the rise kinetics remain in the instrument limited time range (2 microseconds or less) during the first 8-12 flashes. Treating PS II preparations with 800 mM CaCl2 removes the 16, 24 and 33 kDa proteins with at least 95% inhibition of the steady-state rates of O2 evolution. The additional removal of the 33 kDa polypeptide decreases the rates of oxidation and rereduction of Z, the species responsible for Signal II. Preparations treated with either mono- or divalent salts show a steady-state light-induced increase in Signal II similar to that seen in Tris-washed samples. Such a steady-state increase indicates that the rate of electron transport from water to Z is greatly decreased or blocked. The data are interpreted within a model in which there is an intermediate electron carrier between the O2 evolving complex and Z. PMID- 2988611 TI - Orientation of cytochrome c oxidase molecules in the two populations of reconstituted vesicles resolved by column chromatography on DEAE-Sephacryl. AB - Vesicles reconstituted with bovine heart cytochrome c oxidase and dioleoylphosphatidylcholine can be resolved into two populations by column chromatography in DEAE-Sephacryl (Madden, T.D. and Cullis, P.R. (1984) J. Biol. Chem. 259, 7655-7658). These two fractions (I and II) were treated with two proteases. These are trypsin, which has been found to cleave subunit IV in the M domain of the cytochrome c oxidase molecule, and chymotrypsin, which has been found to cleave subunit III in the C domain. These studies show that fraction I vesicles contain cytochrome c oxidase orientation with the M domain outside, i.e., in the same topology as in submitochondrial particles, while fraction II vesicles contain enzyme molecules with their C domain outside, and thus in the same orientation as in mitochondria. PMID- 2988612 TI - Antimycin binds to a small subunit of the ubiquinol: cytochrome c oxidoreductase. AB - Bovine heart ubiquinol: cytochrome c oxidoreductase in Triton X-100 is split with guanidine into a number of fractions. A new method for measuring antimycin binding is developed using extraction with pentanol of the reversibly bound antimycin. By this method and the normal titration method, antimycin-binding capacity is found in a fraction containing a small subunit with a molecular mass of about 12000. This polypeptide was associated with cytochrome c1 but is probably not the 'hinge protein'. Fractions that contain cytochrome b did not show binding by the pentanol-extraction method. PMID- 2988613 TI - Association of spin-labelled cardiolipin with dimyristoylphosphatidylcholine substituted bovine heart cytochrome c oxidase. A generalized specificity increase rather than highly specific binding sites. AB - The endogeneous lipid of bovine heart cytochrome c oxidase has been replaced by dimyristoylphosphatidylcholine using cholate-mediated exchange. The lipid substituted preparation contained less than 1 mole cardiolipin per mole enzyme and possessed full oxidative activity. The association of spin-labelled cardiolipin with such lipid-substituted cytochrome oxidase preparations has been assayed using ESR spectroscopy. An average relative association constant 5.4 times that for phosphatidylcholine is obtained for cardiolipin. Measurements on preparations with increasing contents of unlabelled cardiolipin, introduced during lipid exchange, reveal that this selectivity corresponds to a generalized increase in specificity for all lipid association sites on the protein. PMID- 2988614 TI - Phase transitions and fatty acid spin label behavior in interdigitated lipid phases induced by glycerol and polymyxin. AB - Glycerol and polymyxin have been shown by X-ray diffraction to induce interdigitated bilayers in phosphatidylcholine (PC) and phosphatidylglycerol (PG), respectively (McDaniel, R.V., et al. (1983) Biochim. Biophys. Acta 731, 97 108; Ranck, J.-L. and Tocanne, J.-F. (1982) FEBS Lett. 143, 175-178). In the present study we have investigated the phase behavior of PC and PG in the presence of glycerol and polymyxin by differential scanning calorimetry and the use of fatty acid spin labels. Interdigitation causes a large increase in the order parameter of a fatty acid spin labeled near the terminal methyl, 16 doxylstearate, so that it was similar to that of a fatty acid labeled much closer to the polar head group region, 5-doxylstearate. Thus interdigitation abolishes the fluidity gradient found in a non-interdigitated bilayer. 16-Doxylstearate may be useful in detecting interdigitation of lipid bilayers caused by other substances. The different samples all went through two transitions on heating or cooling, or both. However, use of the fatty acid spin label showed that the molecular events during these transitions varies for different samples. The results suggested that PC-glycerol freezes from the liquid-crystalline phase into a non-interdigitated gel phase. This subsequently becomes interdigitated upon lowering the temperature a few degrees, in a low enthalpy transition. PG polymyxin shows a similar behavior except that the enthalpy of the non interdigitated gel to interdigitated phase transition is greater and the transition is reversible on heating. Thus on heating PG-polymyxin first goes through a transition from the interdigitated phase to a non-interdigitated gel phase and then, in a separate transition, to the liquid-crystalline phase. This occurs because the fatty acid chains in the presence of polymyxin become too disordered with increase in temperature to maintain the interdigitated state. PG glycerol goes into the interdigitated state less readily than the other mixtures. If cooled rapidly, PG-glycerol freezes into a metastable phase which is more disordered than the interdigitated phase. It goes into the interdigitated phase in an exothermic transition on heating. An increase in fatty acid chain length causes greater steric hindrance to interdigitation but also increases the stabilizing energy gained by interdigitation. PMID- 2988615 TI - Calcium sequestration activity in rat liver microsomes. Evidence for a cooperation of calcium transport with glucose-6-phosphatase. AB - Mechanisms regulating the energy-dependent calcium sequestering activity of liver microsomes were studied. The possibility for a physiologic mechanism capable of entrapping the transported Ca2+ was investigated. It was found that the addition of glucose 6-phosphate to the incubation system for MgATP-dependent microsomal calcium transport results in a marked stimulation of Ca2+ uptake. The uptake at 30 min is about 50% of that obtained with oxalate when the incubation is carried out at pH 6.8, which is the pH optimum for oxalate-stimulated calcium uptake. However, at physiological pH values (7.2-7.4), the glucose 6-phosphate-stimulated calcium uptake is maximal and equals that obtained with oxalate at pH 6.8. The Vmax of the glucose 6-phosphate-stimulated transport is 22.3 nmol of calcium/mg protein per min. The apparent Km for calcium calculated from total calcium concentrations is 31.9 microM. After the incubation of the system for MgATP dependent microsomal calcium transport in the presence of glucose 6-phosphate, inorganic phosphorus and calcium are found in equal concentrations, on a molar base, in the recovered microsomal fraction. In the system for the glucose 6 phosphate-stimulated calcium uptake, glucose 6-phosphate is actively hydrolyzed by the glucose-6-phosphatase activity of liver microsomes. The latter activity is not influenced by concomitant calcium uptake. Calcium uptake is maximal when the concentration of glucose 6-phosphate in the system is 1-3 mM, which is much lower than that necessary to saturate glucose-6-phosphatase. These results are interpreted in the light of a possible cooperative activity between the energy dependent calcium pump of liver microsomes and the glucose-6-phosphatase multicomponent system. The physiological implications of such a cooperation are discussed. PMID- 2988616 TI - Modulation of prolactin binding sites in vitro by membrane fluidizers. Effects on male prostatic and female hepatic membranes in alcohol-fed rats. AB - The objectives of this study were (i) to determine if in vivo administration of ethanol to rats produced changes in apparent lipid fluidity and prolactin binding capacity of male prostatic and female hepatic membranes and (ii) to compare the effects of membrane fluidizers (aliphatic alcohols) in vitro on prolactin binding of prostatic and hepatic membranes in control and alcohol-fed animals. In vitro ethanol has been shown by us previously to increase prolactin receptor levels presumably by unmasking cryptic prolactin receptors. The degree of fluidization was monitored by a fluorescence polarization method using 1,6-diphenylhexatriene. Adult male and female rats were given either water or 4% ethanol as the sole source of drinking fluid for a period of 6 weeks. No significant changes in plasma prolactin were observed between control and ethanol-treated groups of either sex. However, the microviscosity parameter, inversely related to lipid fluidity, was increased approx. 34% and 40%, respectively, in male prostatic and female rat hepatic membranes after ethanol feeding. Furthermore, 125I-prolactin binding capacity was decreased approx. 30% and 26%, respectively, in prostatic and hepatic membranes of alcohol fed animals. In vitro treatment with aliphatic alcohols had no effect on either microviscosity or prolactin binding in hepatic or prostatic membranes from ethanol-fed rats, but both fluidized and increased prolactin binding in the same membrane preparations from control rats. Our observations are consistent with the direct relationship between membrane fluidity and prolactin receptor levels. The changes in prostatic and hepatic membranes after alcohol feeding, namely decreased prolactin receptor levels, decreased fluidity and increased resistance to the fluidizing effects of in vitro aliphatic alcohols may reflect a fundamental membrane defect. PMID- 2988617 TI - The activity of membranes reconstituted from HVJ envelope proteins and lipids to induce hemolysis and fusion between liposomes and erythrocytes. AB - A simple method for preparation of lipid-free envelope proteins (HN protein and F protein) of HVJ (Sendai virus) was developed. Reconstituted 'envelopes' were then prepared from envelope proteins and various lipids by the detergent dialysis method, and the activity to induce hemolysis and fusion between liposome and erythrocyte was studied. Lipid-free envelope protein aggregates could induce hemolysis and liposome-erythrocyte fusion. The activity was however greatly augmented by incorporation of envelope proteins into membrane of viral total lipids. Hemolytic and fusogenic activity was somewhat augmented by incorporation of envelope proteins into dipalmitoylphosphatidylcholine/cholesterol (1:1, molar ratio) and dimyristoylphosphatidylcholine/cholesterol (1:1), though the augmentation was lower than that observed with viral total lipids. When 'envelopes' were reconstituted with the proteins and viral total lipids supplemented with phosphatidylethanolamine, two kinds of 'envelopes' were prepared; one was permeable to Dextran (Mr 75000) and hemolytic, and the other was impermeable to Dextran and nonhemolytic. The latter acquired hemolytic activity after subjection to freezing and thawing, and its barrier function was lost concomitantly. The study suggests that envelope proteins (HN protein and F protein) could function without lipids but their activity was greatly influenced by not only the composition of additional lipids but also mode of arrangement of components on the reconstituted membranes. PMID- 2988618 TI - Thiocyanate and nitrite inhibit proton translocation in gastric mucosa. AB - Isolated frog gastric mucosa was used to study the separation of formation of protons (or their precursors) from proton translocation by using various inhibitors. Both thiocyanate (SCN-) and nitrite (NO2-) inhibit the acid secretion in spontaneously secreting mucosa. The inhibition is reversed when the inhibitor is removed such that the excess acid secreted above baseline in the 'off'-period compensates for the amount inhibited in the 'on'-period. Both agents also inhibit the effect on acid secretion of pulse stimulation with histamine though to a lesser extent. Upon removal of the inhibitor, the total amount of acid secreted in excess of basal is equal to that observed with histamine alone. Likewise, metiamide, an H2-antagonist, also inhibits acid secretion with or without histamine. However, in contrast to SCN- and NO2-, removal of this inhibitor is without effect on the acid-secretion rate. These results indicate that both SCN- and NO2- inhibit the proton translocation rather than the formation of protons or their precursors as is the case with metiamide. PMID- 2988619 TI - Asymmetric orientation of amino groups in the alpha-subunit and the beta-subunit of (Na+ + K+)-ATPase in tight right-side-out vesicles of basolateral membranes from outer medulla. AB - The orientation of amino groups in the membrane in the alpha- and beta-subunits of (Na+ + K+)-ATPase was examined by labeling with Boldon-Hunter reagent, N succinimidyl 3-(4-hydroxy,5-[125I]iodophenyl)propionate), in right-side-out vesicles or in open membrane fragments from the thick ascending limbs of the Henles loop of pig kidney. Sealed right-side-out vesicles of basolateral membranes were separated from open membrane fragments by centrifugation in a linear metrizamide density gradient. After labeling, (Na+ + K+)-ATPase was purified using a micro-scale version of the ATP-SDS procedure. Distribution of label was analyzed after SDS-gel electrophoresis of alpha-subunit, beta-subunit and proteolytic fragments of alpha-subunit. Both the alpha- and the beta-subunit of (Na+ + K+)-ATPase are uniformly labeled, but the distribution of labeled residues on the two membrane surfaces differs markedly. All the labeled residues in the beta-subunit are located on the extracellular surface. In the alpha subunit, 65-80% of modified groups are localized to the cytoplasmic surface and 20-35% to the extracellular membrane surface. Proteolytic cleavage provides evidence for the random distribution of 125I-labeling within the alpha-subunit. The preservation of (Na+ + K+)-ATPase activity and the observation of distinct proteolytic cleavage patterns of the E1- and E2-forms of the alpha-subunit show that the native enzyme structure is unaffected by labeling with Bolton-Hunter reagent. Bolton-Hunter reagent was shown not to permeate into sheep erythrocytes under the conditions of the labeling experiment. The data therefore allow the conclusion that the mass distribution is asymmetric, with all the labeled amino groups in the beta-subunit being on the extracellular surface, while the alpha subunit exposes 2.6-fold more amino groups on the cytoplasmic than on the extracellular surface. PMID- 2988620 TI - Involvement of sulfhydryl groups in the inhibition of brain (Na+ + K+)-ATPase by pyrithiamin. AB - Brain (Na+ + K+)-ATPase was protected by low concentrations of GSH from the inhibitory effect of pyrithiamin. The possible involvement of sulfhydryl groups in the inhibition was then studied by comparing the effect of pyrithiamin with that of N-ethylmaleimide on the enzyme. The treatment of rat brain (Na+ + K+) ATPase with thesee inhibitors caused a significant decrease in reactivity of the enzyme to N-ethyl[3H]maleimide. N-Ethylmaleimide, like pyrithiamin, inhibited the partial reactions of (Na+ + K+)-ATPase system in parallel with the inhibition of the overall reaction. An SDS-polyacrylamide gel electrophoresis procedure indicated that pyrithiamin and N-ethylmaleimide inhibited Na+-dependent phosphorylation of the alpha(+) form of rat brain (Na+ + K+)-ATPase more than that of alpha, though the selectivity for the alpha(+) seemed to be higher with the former inhibitor than in the latter. The treatment also decreased sensitivity of the enzyme to ouabain inhibition. However, pyrithiamin- and N-ethylmaleimide induced inactivations of the enzyme differed in the efficacy of GSH for protection and in the effect of the kind of ligands present during the reaction. Furthermore, pyrithiamin did not appear to interact directly with sulfhydryl groups, but caused the formation of disulfide in bovine brain (Na+ + K+)-ATPase. In contrast to N-ethylmaleimide, pyrithiamin did not affect the sulfhydryl enzymes such as alcohol dehydrogenase and L-alanine dehydrogenase. It is concluded that pyrithiamin modifies the functional sulfhydryl groups of brain (Na+ + K+)-ATPase in a way different from N-ethylmaleimide and causes a structural change and inactivation of the enzyme. PMID- 2988621 TI - The mechanism for inhibition of gastric (H+ + K+)-ATPase by omeprazole. AB - Omeprazole was found to inhibit the K+-stimulated ATPase activity of the gastric (H+ + K+)-ATPase in parallel with the K+-stimulated p-nitrophenylphosphatase activity and the phosphoenzyme formation. The degree of inhibition of ATPase activity was directly correlated to the amount inhibitor bound to the enzyme preparation down to about 15% of the control enzyme activity. The acid-decomposed form of omeprazole, i.e. the inhibitory form, was found to react with and bind to sulfhydryl groups within the (H+ + K+)-ATPase preparation with close to a 1:1 stoichiometry. beta-Mercaptoethanol, when added beforehand and in a 10-fold excess of omeprazole, completely prevented binding of the inhibitor and its inhibition of the enzyme. In the presence of beta-mercaptoethanol two different reaction products could be detected in addition to omeprazole; the reduced form of omeprazole (H 168/22), and a product formed between beta-mercaptoethanol and a decomposition product, generated from omeprazole. Under those conditions neither inhibition nor binding was obtained, indicating that none of these three compounds was the inhibitor. Rather, the compound generated from omeprazole and reacting rapidly with either beta-mercaptoethanol or the -SH groups of the enzyme was the likely inhibitor compound. In order to reverse already established inhibition higher concentrations of beta-mercaptoethanol were needed than for protection indicating two different reaction pathways for protection and reversal by beta-mercaptoethanol. The reversal reaction was explained by a two-step reaction; in the first step the bound inhibitor was exchanged for a beta mercaptoethanol molecule resulting in formation of compound H 168/22 and a mixed disulfide between the enzyme and beta-mercaptoethanol. In the second step, attack of another beta-mercaptoethanol molecule results in liberation of active enzyme and generation of the disulfide form of beta-mercaptoethanol. This hypothesis was substantiated by the fact that when 1 mM beta-mercaptoethanol was added to inhibited enzyme the radiolabel was partially displaced, without any change in the concentration of modified -SH groups. PMID- 2988622 TI - The locus of nucleotide specificity in the reaction mechanism of (Na+ + K+) ATPase determined with ATP and GTP as substrates. AB - ATP and GTP have been compared as substrates for (Na+ + K+)-ATPase in Na+ activated hydrolysis, Na+-activated phosphorylation, and the E2K----E1K transition. Without added K+ the optimal Na+-activated hydrolysis rates in imidazole-HCl (pH 7.2) are equal, but are reached at different Na+ concentrations: 80 mM Na+ for GTP, 300 mM Na+ for ATP. The affinities of the substrates for the enzyme are widely different: Km for ATP 0.6 microM, for GTP 147 microM. The Mg-complexed nucleotides antagonize activation as well as inhibition by Na+, depending on the affinity and concentration of the substrate. The optimal 3-s phosphorylation levels in imidazole-HCl (pH 7.0) are equally high for the two substrates (3.6 nmol/mg protein). The Km value for ATP is 0.1-0.2 microM and for GTP it ranges from 50 to 170 microM, depending on the Na+ concentration. The affinity of Na+ for the enzyme in phosphorylation is lower with the lower affinity substrate: Km (Na+) is 1.1 mM with ATP and 3.6 mM with GTP. The GTP-phosphorylated intermediate exists, like the ATP-phosphorylated intermediate, in the E2P conformation. Addition of K+ increases the optimal hydrolytic activity 30-fold for ATP (at 100 mM Na+ + 10 mM K+) and 2-fold for GTP (at 100 mM Na+ + 0.16 mM K+). K+ greatly increases the Km values for both substrates (to 430 microM for ATP and 320 microM for GTP). Above 0.16 mM K+ inhibits GTP hydrolysis. GTP does not reverse the quenching effect of K+ on the fluorescence of the 5-iodoacetamidofluorescein-labeled enzyme. ATP fully reverses this effect, which represents the transition from E1K to E2K. Hence GTP is unable to drive the E2K----E1K transition. PMID- 2988623 TI - Reconstitution of membrane receptor systems. AB - This review makes an attempt to summarize the present status of the field of receptor reconstitution. First a general discussion on the problem of receptor to effector coupling is discussed with an emphasis on the approaches used to solubilize, purify and reconstitute receptors with their respective biochemical effectors. Two categories of receptors have thus far been studied in great detail: (1) receptors linked to ion channels best represented by the nicotinic acetylcholine receptor and (2) receptors linked to adenylate cyclase. Through a detailed discussion of these two receptor systems the reader should get an idea of where the field of receptor reconstitution is headed. Only in the beta adrenergic-receptor-dependent adenylate cyclase have the receptor and the effector systems been completely separated, purified and reconstituted. Therefore, a detailed discussion on that system occupies a very significant portion of this article. A summary of the state-of-the-art on a number of other receptor systems is also given in the last part of the review. PMID- 2988624 TI - Specificity of lipid-protein interactions as determined by spectroscopic techniques. PMID- 2988625 TI - Comparative analysis of the human and feline c-sis proto-oncogenes. Identification of 5' human c-sis coding sequences that are not homologous to the transforming gene of simian sarcoma virus. AB - Feline and human genetic sequences, homologous to the v-sis gene of simian sarcoma virus, have been isolated from cosmid gene libraries and characterized by restriction endonuclease analysis. Comparison of the two loci revealed their related structural organization. In both loci, similar unique genetic sequences were found upstream of the v-sis homologous region and these hybridized to a 4.2 kbp c-sis transcript in human lung tumor cells. These data establish and map as yet unidentified coding sequences at the 5' part of the c-sis proto-oncogene of both species. PMID- 2988626 TI - Chromatin structure of the 87A7 heat-shock locus during heat induction and recovery from heat shock. AB - We have examined the chromatin organization of the 87A7 heat-shock locus (which contains two hsp 70 genes transcribed in opposite orientation) as a function of the time of heat induction and during the course of recovery from heat shock. Our studies show that both induction and recovery from heat shock are accompanied by highly specific alterations in the nucleoprotein structure of this locus. Moreover, these changes parallel the transcriptional activity of the hsp 70 heat shock genes. We have also examined the effect of inhibitors of transcription and translation. Cycloheximide, an inhibitor of translation, blocks both the attenuation of the heat-shock response (which occurs after a long-term incubation at elevated temperatures) and the re-establishment of the pre-induced chromatin organization of the locus during recovery from heat shock. Actinomycin D, an inhibitor of transcription, prevents some but not all of the alterations in chromatin structure which normally accompany heat induction. PMID- 2988627 TI - Displacement synthesis of globin complementary DNA: evidence for sequence amplification. AB - We have examined a cDNA displacement synthesis procedure in which the extent of precursor incorporation and the unusual kinetics of displacement synthesis suggest a unique replicative form of DNA and the occurrence of multiple rounds of displacement synthesis, leading to amplification of mRNA sequences. Globin double stranded DNA containing a hairpin loop was extended by the addition of a homopolymer to the 3' end. This was followed by displacement synthesis with the Klenow fragment of DNA polymerase I that was primed by an oligonucleotide hybridized to the homopolymer. Thus, the hairpin cDNA was copied to form an open duplex with an inverted repetition of globin sequences. These molecules can then serve as templates for additional synthesis which would be primed from oligomers bound the homopolymer. Globin cDNA sequences appear to be amplified 10-fold or more by this procedure. Globin cDNA obtained by displacement synthesis was similar in size to the original template. However, displaced molecules associate to the extent that they are not readily resolved by electrophoresis or sedimentation under nondenaturing conditions. Restriction endonuclease digests of 32P-labeled displaced strands gave fragment patterns similar to rabbit globin cDNA hairpin molecules. S1 nuclease studies demonstrated that displaced complexes and replication intermediates are partially single stranded, which might account for their aggregation properties. PMID- 2988628 TI - Glucocorticoid modulation of collagenase expression in human skin fibroblast cultures. Evidence for pre-translational inhibition. AB - Glucocorticoids inhibit collagenase accumulation in the medium of human skin explant cultures. To examine the mechanism for this process, skin fibroblasts were placed in serum-free medium containing various steroids. Dexamethasone produced a dose-dependent inhibition of trypsin-activatable collagenase in the culture medium with maximal inhibition of approx. 85% at 10(-6) M. Dexamethasone failed to inhibit collagenase activity directly. The decrease in activity in the medium was paralleled by a decrease in immunoreactive protein, suggesting inhibition of enzyme synthesis. The specificity of the effect was shown in two ways. At 10(-6) M steroid, only dexamethasone and hydrocortisone were inhibitory; estradiol, progesterone and testosterone produced less than 10% inhibition. In biosynthetic studies, exposure to 10(-7) M dexamethasone for 24 h produced approx. 50% inhibition of collagenase synthesis but caused no greater than 10% inhibition of total protein synthesis. The T1/2 for achieving the effect was approx. 16 h after initial exposure to dexamethasone. These kinetics were parallel to the inhibition caused by actinomycin D and cordycepin, two inhibitors of transcription, but were longer than that caused by cycloheximide (T 1/2 less than 3 h). To examine this process, cells were cultured in the presence or absence of 10(-6) M dexamethasone prior to harvesting mRNA for cell-free translation. In each case the inhibition or enzyme activity in the intact cells was paralleled by a reduction in translatable collagenase mRNA from the same cells. At the same time, there was no significant inhibition of total protein translation by the steroid. These data suggest that glucocorticoids regulate collagenase synthesis at a pre-translational level, possibly through inhibition of transcription. PMID- 2988629 TI - Function and structure of microvirid phage alpha 3 genome. DNA sequence of H gene and properties of missense H mutant. AB - The nucleotide sequence of wild-type alpha 3 H gene and its surrounding region was determined and compared with those of phi X174 and G4. The corresponding DNA regions in double mutants amJH22, amJH69 and amJH76 were also sequenced and their missense mutation sites located. A phage strain missH22 having a single missense mutation in gene H was constructed by replacing the J region of amJH22 in vitro with the wild-type DNA. Like amJH22, the missense mutant coded for H protein with aberrant electrophoretic mobility, but formed normal plaques on suppressor deficient Escherichia coli. Heat stability, plating efficiency on certain hosts and rate of eclipse were higher in strain missH22 than in wild-type phage. PMID- 2988630 TI - Preparation and properties of a single-sited fragment from the C-terminal domain of human transferrin. AB - A single-sited iron-binding fragment of human transferrin has been obtained by thermolysin cleavage of the protein, selectively loaded with iron in the C terminal binding site, in a urea-containing buffer. The fragment contains carbohydrate, and hence derives from the C-terminal half of transferrin. Its metal-binding site accepts Fe3+ and Cu2+ with bicarbonate as accompanying anion, but only Fe3+ with oxalate as anion. EPR spectroscopic properties of the fragment are similar to those of the corresponding site in the intact protein. However, iron-binding by the fragment is weaker than by the C-terminal site of the intact protein, particularly at low pH, suggesting that overall as well as local protein conformation influences the metal-binding functions of the site. PMID- 2988631 TI - An alkaline thiol proteinase in the liver mitochondria of bullfrog, Rana catesbeiana. AB - The mitoplasts were prepared from bullfrog (Rana catesbeiana) liver mitochondria by treatment with digitonin and were then separated into the matrix and inner membrane fractions. The matrix fraction thus obtained was free of lysosomal contaminations and exhibited a distinct proteinase activity. pH dependency of the matrix proteinase activity measured in the presence and absence of iodoacetamide revealed that the matrix contained at least two kinds of proteinase, a major alkaline thiol proteinase having an optimal pH at 8.5 and a minor neutral proteinase having an optimal pH at 7.5. The major matrix proteinase activity was strongly inhibited by leupeptin, chymostatin, antipain and E64-C, an inhibitor of Ca2+-dependent thiol proteinase, while it was scarcely affected by diethylpyrocarbonate. The activity was also inhibited by DTNB and p chloromercuribenzoate. Addition of hydrocarbon compounds such as ethylene glycol, glycerol, Triton X-100 and poly (ethylene glycol) to the reaction mixture was found to decrease the matrix proteinase activity. Neither cytochrome c nor glutamate dehydrogenase was hydrolyzed when subjected to the matrix proteinase activity in vitro. On the other hand, cytochrome c oxidase was effectively hydrolyzed, and the enzyme associated with the mitochondrial innermembrane fragments was partially hydrolyzed by the major matrix proteinase activity. PMID- 2988632 TI - Developmental differences in activation of cholinephosphate cytidylyltransferase by lipids in rabbit lung cytosol. AB - Lung cytosolic cholinephosphate cytidylyltransferase is activated by lipids. We examined the lipid activation pattern as a function of development in rabbit lung from 27 days gestation through term (31 days) and in the adult. The enzyme in both the fetal and adult cytosol was dependent on lipids for activity. Extraction of the cytosol with acetone/butanol virtually abolished cytidylyltransferase activity, but the activity could be restored on addition of lipids extracted with chloroform/methanol from additional cytosol. Cytosolic phospholipids from the fetal lung reactivated cytidylyltransferase but both neutral lipids and phospholipids from the adult were required. The lipids had the same effect on cytidylyltransferase activity in delipidated cytosol from either the fetus or adult so the difference in activation pattern was attributable to the lipids rather than the protein. There was a shift from the fetal to the adult lipid activation pattern as development progressed. Further, there was a significant correlation between cytidylyltransferase activities in intact cytosols from developing lung and activities in delipidated cytosol in the presence of lipids from the same animals. Although these data suggest that lipids regulate cytosolic cytidylyltransferase activity in developing lung their physiological significance remains to be established. PMID- 2988633 TI - Phosphatidylglycerol stimulates cholinephosphate cytidylyltransferase activity and phosphatidylcholine synthesis in type II pneumocytes. AB - Phosphatidylcholine synthesis in type II pneumocytes is stimulated by inclusion of phosphatidylglycerol and other phospholipids in the culture medium (Gilfillan, A.M., Chu, A.J. and Rooney, S.A. (1984) Biochim. Biophys. Acta 794, 269-273). We have now examined the effect of phosphatidylglycerol in the medium on enzymes of de novo phosphatidylcholine synthesis in adult rat type II cells. Activities of choline kinase, cholinephosphate cytidylyltransferase and cholinephosphotransferase in homogenates of whole lung and type II cells were generally similar. Phosphatidate phosphatase activity in type II cells, however, was only 16% that in whole lung. Addition of phosphatidylglycerol (10 microM) to the culture medium had no effect on choline kinase, cholinephosphotransferase or phosphatidate phosphatase activities in type II cells but it increased the activity of cholinephosphate cytidylyltransferase by 56%. Since it is known that cholinephosphate cytidylyltransferase is stimulated in vitro by addition of phospholipids to the assay mixture, we also measured its activity in the presence of sufficient phosphatidylglycerol (1.1 mM) to maximally stimulate in vitro. Even under these conditions cholinephosphate cytidylyltransferase activity in type II cells cultured in the presence of phosphatidylglycerol was 32% greater than in control cells. These data show that the stimulatory effect of phospholipid in the culture medium on phosphatidylcholine synthesis in type II cells is mediated by increased cholinephosphate cytidylyltransferase activity. The mechanism of increased cytidylyltransferase activity remains to be elucidated but it is not due to direct in vitro activation by the phospholipid. PMID- 2988634 TI - Recombinant models of lipoproteins. Apolipoprotein A I/phosphatidylcholine/cholesterol complexes formed in a 2-chloroethanol-water mixture. AB - Apolipoprotein A-I can spontaneously associate with phosphatidylcholine and cholesterol in 2-chloroethanol-water mixture. It was demonstrated, using a spin label technique, that dissolved molecules participate in complex formation. The apolipoprotein A-I/phosphatidylcholine/cholesterol complexes were isolated by gel chromatography. Complexes of three types were prepared and characterized: type A, large heterogeneous aggregates with molecular weight 600 000, sedimentation coefficient 10 S and the following molar composition - protein/phosphatidylcholine/cholesterol, 1:(70-100):(10-12); types B and C, with weight average molecular weights 140 000 and 110 000, average sedimentation coefficients 3.6 S and 1.7 S, respectively. Both types have the same molar composition - protein/phosphatidylcholine/cholesterol, 1:25:8. The dissimilar sedimentation coefficients between complexes B and C may be explained by the difference in the monomer/tetramer ratio (monomer molecular weight 50 000). The spin label sn-1-O-stearoyl-2-O-9'-spiro(4'',4''-dimethyloxazolidine-3''-oxyl) heptadecanoylglycero-3-phosphocholine introduced into the complexes A and B showed different thermal properties of these complexes, which may be due to differences in the lipid-protein interactions. PMID- 2988635 TI - The biosynthesis of leukotriene B4, the predominant lipoxygenase product in rabbit alveolar macrophages, is enhanced during immune activation. AB - Rabbit alveolar macrophages synthesize prostaglandins in response to various stimuli. We have previously shown that prostaglandin production was decreased in immunologically activated (live Bacillus Calmette-Guerin (BCG)-injected) animals. (Hsueh, W., Lamb, R. and Gonzalez-Crussi, F. (1982) Biochim. Biophys. Acta 710, 406-414). In the present study, we examined the lipoxygenase products of alveolar macrophages from normal and immunologically activated rabbits injected intravenously with live BCG or complete Freund's adjuvant. We found: unstimulated lung macrophages produced no detectable leukotrienes; the predominant lipoxygenase product upon stimulation was leukotriene B4; alveolar macrophages did not significantly degrade leukotriene B4 into its 20-hydroxy derivative, and the total degradation of leukotriene B4 during 90 min of incubation was minimal; the production of leukotriene B4 reached the peak at 30 min after A23187 stimulation, while zymosan caused a much slower and smaller release; following stimulation, immunologically activated lung macrophages produced more leukotriene B4 than resident macrophages. It is possible that the increased leukotriene B4 production in immunologically activated lung macrophages was related to the immunoregulatory function of this substance, such as enhancing cytotoxicity, interferon production and proliferation of suppressor-cytotoxic T cells. PMID- 2988636 TI - Angiotensin II stimulates receptor-mediated uptake of LDL by bovine adrenal cortical cells in primary culture. AB - Bovine adrenal cells were isolated from the subcapsular region of the gland to obtain cultures enriched in cells of the zona glomerulosa. The cells kept in primary cultures were shown to respond to angiotensin II and adrenocorticorticotropin (ACTH) by a significant increase in aldosterone production. These primary adrenal cultures were used to study the effect of angiotensin II on LDL metabolism. Addition of angiotensin II for 48 h to the culture medium resulted in a 200-300% increase in LDL metabolism, and the lowest effective concentration was 10(-8) -10(-9) M. The angiotensin II effect became evident after 12-16 h of incubation. To compare the metabolism of the 125I labeled protein moiety to that of cholesteryl ester of LDL, the lipoprotein was labeled also with cholesteryl linoleyl ether, a nonhydrolyzable analog of cholesteryl ester. Under basal conditions and in the presence of angiotensin II or ACTH the ratio of [3H]cholesteryl linoleyl ether to 125I indicate some preferential uptake of the cholesteryl ester moiety. Stimulation of specific LDL binding at 4 degrees C and LDL metabolism at 37 degrees C by 10(-7) M angiotensin II occurred at all concentrations of LDL studied. Linearization of the kinetic data showed that angiotensin II increased the LDL receptor number significantly but not the affinity of the LDL receptor for its ligand. The present findings indicate that in analogy to ACTH, angiotensin II can influence receptor-mediated uptake of LDL by adrenal cortical cells. It remains to be shown whether the angiotensin II effect on LDL metabolism is limited to adrenal cells or will affect other cells which express the angiotensin II receptor. PMID- 2988637 TI - Metabolism of eicosapentaenoic acid by aorta: formation of a novel 13 hydroxylated prostaglandin. AB - We have investigated the metabolism by fetal calf aorta of eicosapentaenoic acid (20:5) and docosahexaenoic acid (22:6), two polyunsaturated fatty acids found in high concentrations in marine oils. The major product formed from 20:5 by particulate fractions from fetal calf aorta is delta 17-6-oxoprostaglandin F1 alpha. In addition, we detected a novel isomer of delta 17-6-oxoprostaglandin F1 alpha, in which a hydroxyl group is present in the 13-position instead of the 15 position. Eicosapentaenoic acid is also converted to 12-hydroxy-5,8,10,14 heptadecatetraenoic acid as well as to five monohydroxy isomers with hydroxyl groups present in the 11, 12, 14, 15, and 18 positions. Although 20:5 was metabolized at about one-third the rate of arachidonic acid (20:4), greater amounts of monohydroxy fatty acids, the major one being the 11-hydroxy metabolite, were formed from 20:5. Unlike 20:5, 22:6 was not metabolized to any detectable products by fetal calf aorta, but both of these polyunsaturated fatty acids inhibited the oxygenation of 20:4 by cyclooxygenase from aorta with IC50 values of 4.1 microM (22:6) and 15 microM (20:5). These results suggest that 20:5 has a high affinity for cyclooxygenase, but that the intermediate 11-oxygenated intermediate has a lower affinity than the corresponding intermediate from 20:4, resulting in a greater loss of substrate after a single oxygenation. The formation of oxygenation products from both 20:4 and 20:5 was inhibited by 13 hydroperoxy-9,11-octadecadienoic acid (13hp-18:2). The IC50 values for inhibition of cyclooxygenase products by 13hp-18:2 were about twice as high as those for inhibition of prostacyclin synthase products. Consequently, there was little diversion of prostaglandin endoperoxides to other prostaglandins in the presence of 13hp-18:2. PMID- 2988638 TI - Characterisation of heterologous and homologous low-density lipoprotein binding to apolipoprotein B,E receptors on porcine adrenal cortex membranes: enhanced binding of trypsin-modified human low-density lipoprotein. AB - The characteristics of the binding of homologous and heterologous (human) LDL to membrane preparations from porcine adrenal cortex have been determined. The membranes displayed a single class of high-affinity, saturable binding site for both 125I-labelled porcine and human LDL, which was dependent on divalent cations, in addition to a low-affinity, non-saturable component(s). Porcine LDL displaced both 125I-labelled porcine and 125I-labelled human LDLs from the high affinity binding site more effectively than human LDL, reflecting the lower Kd, (13.2 micrograms/ml) for porcine than human (Kd 19.2 micrograms/ml) LDL. These values are comparable to those obtained for half-maximal binding of human and bovine LDLs in a bovine adrenocortical membrane system (Kovanen, P.T., Basu, S.K., Goldstein, J.L. and Brown, M.S. (1979) Endocrinology 104, 610-616). Tryptic modification of porcine LDL (T-LDL) diminished its ability to compete with 125I labelled native LDL for the high-affinity binding site; in contrast, 125I labelled porcine T-LDL showed an elevated receptor affinity (Kd 9.7 micrograms/ml) and was more efficiently displaced by its unlabelled counterpart than by native porcine LDL. Tryptic treatment of human LDL similarly increased its binding affinity (Kd 8.3 micrograms/ml), although in this case, the unlabelled T-LDL displaced not only 125I-labelled human T-LDL but also 125I labelled human LDL from the high-affinity site more effectively than native LDL. We conclude that (i) porcine adrenocortical membranes possess binding sites specific for LDL and resembling the apolipoprotein B,E receptors already demonstrated in murine, bovine and human adrenal cortex; (ii) tryptic modification of porcine LDL may remove or destroy segments of apolipoprotein B100 which contribute to receptor recognition sites on the surface of the particle; (iii) trypsinised porcine LDL may interact with the membrane binding site by a mechanism differing from that by which native LDL binds, and (iv) trypsinisation of human LDL may cleave or remove species-specific segments of the B100 protein at or close to the receptor recognition site(s) on the particle, thus decreasing structural differences between porcine and human LDL, and thereby enhancing its binding affinity for the porcine receptor. PMID- 2988639 TI - Utilization of exogenous glycerophosphodiesters and glycerol 3-phosphate by inositol-starved yeast, Saccharomyces uvarum. AB - Inositol-starved Saccharomyces uvarum cells hydrolyse exogenous glycerophosphodiesters to glycerol 3-phosphate and the corresponding alcohol. Glycerophosphodiesterase activity is highest with glycerophosphoinositol as the substrate, followed by glycerophosphoethanolamine and glycerophosphocholine; the artificial substrate for phosphodiesterases, bis-p-nitrophenylphosphate,is hydrolysed at a similar rate as compared with glycerophosphoinositol. Competition experiments suggest that distinct phosphodiesterases are involved in the hydrolysis of the respective substrates. An Mg2+-dependent glycerophosphate phosphohydrolase with a pH-optimum around neutral cleaves glycerol 3-phosphate to glycerol and orthophosphate. The latter is taken up into cells without first entering the pool of orthophosphate present in the growth medium. Accessibility to substrates with whole cells, adhesion of enzymes to spheroplasts, and solubilization of enzymes by treatment of whole cells with Triton X-100 under mild conditions suggest that phosphodiesterases and glycerol-3-phosphate phosphohydrolase are loosely associated with the outer side of the yeast plasma membrane. Enzyme activities are only marginal in inositol-supplemented cells, but are derepressed not only by inositol deficiency, but also by starvation of orthophosphate. PMID- 2988640 TI - Analysis of the effects of fatty acids and related compounds on the synthesis of phosphatidylcholine in lymphocytes. AB - Phosphatidylcholine synthesis in cultured bovine lymphocytes is stimulated by cis unsaturated fatty acids. This stimulation is correlated with an activation of the enzyme cytidyltransferase (EC 2.7.7.15) and its apparent translocation from the cytosol to the membrane/particulate of cells. In addition, these agents increase the levels of cytidine diphosphocholine - a product of the cytidyltransferase reaction and a precursor to phosphatidylcholine. Retinoic acid and 5,8,11,14 eicosatetraynoic acid both activate cytidyltransferase activity and raise cytidine diphosphocholine levels, yet they are ineffective as stimulators of overall phosphatidylcholine synthesis. The effects of all of these lipids are reversed by the delayed addition of bovine serum albumin. The data point to the view that cytidyltransferase activation is required but is not sufficient for stimulation of phosphatidylcholine synthesis: regulation at another step is suggested. PMID- 2988641 TI - Leukotriene A4-hydrolase activity in guinea pig and human liver. AB - Guinea pig and human liver homogenates transformed leukotriene A4 into leukotriene B4. In both species, the enzymatic activity was recovered in the 105000 X g supernatant, and it was found to be susceptible to heat treatment (56 degrees C, 1 h). Digestion with a proteolytic enzyme also resulted in loss of enzymatic activity. The formation of leukotriene B4 was pH-dependent, with an optimum between pH 7 and pH 8.5. In addition, two other organs from the guinea pig, lungs and kidneys, contained leukotriene A4-hydrolase activity. The identity of leukotriene B4 was ascertained by high-performance liquid chromatography, ultraviolet spectrometry, gas chromatography-mass spectrometry and bioassay. We have recently demonstrated the presence of leukotriene A4-hydrolase activity in mammalian plasma (Fitzpatrick et al. (1983) Proc. Natl. Acad. Sci. USA 80, 5425 5429). The results of the present study suggest several possible origins of this plasma leukotriene A4 hydrolase. PMID- 2988643 TI - A temperature-sensitive Chinese hamster ovary cell mutant pleiotropically defective in protein export. AB - We have developed a new selection procedure for mammalian cell mutants defective in protein export by the use of diphtheria toxin, and devised a new screening method for defective protein secretion using nitrocellulose membranes. By the combination of these procedures, we have isolated a temperature-sensitive mutant clone of Chinese hamster ovary cells which shows a pleiotropic defect in protein export. This mutant, designated DS28-6, is temperature-sensitive for growth. Secretion of a series of proteins is markedly inhibited at the non-permissive temperature. These proteins seem to be normally synthesized and accumulated within the cell at the non-permissive temperature and secreted upon shift down to the permissive temperature. When this mutant is infected with vesicular stomatitis virus, oligosaccharide processing of G-protein is arrested at an endoglycosidase-H-sensitive stage at the non-permissive temperature. The lesion of this mutant appears to be in the endoplasmic reticulum or the cis Golgi or both. PMID- 2988642 TI - Inhibition of lipid peroxidation by heme-nonapeptide derived from cytochrome c. AB - Heme-nonapeptide, derived from cytochrome c, inhibited both the NADPH- and NADH dependent lipid peroxidation of brain microsomes but, in the case of liver microsomes, this inhibitory effect manifested itself in the presence of SKF-525A (a specific blocker of cytochrome P-450) only. Heme-nonapeptide prevented the transient accumulation of lipid peroxides in microsomes during lipid peroxidation. The oxygen consumption of microsomes in the presence of NADPH or NADH was stimulated by heme-nonapeptide. From these results we concluded that, in vitro, there are two independent mechanisms of lipid peroxidation in liver microsomes. It is suggested that, in vivo, the heme-peptide-sensitive mechanism, observed in brain microsomes, is more important. PMID- 2988644 TI - Cholesterol modulation of beta-adrenergic receptor characteristics. AB - Cholesterol, a major structural component of plasma membranes, has a profound influence on cell surface receptor characteristics and on adenylate cyclase activity. beta-Adrenergic receptor number, adenylate cyclase activity, and receptor-cyclase coupling were assessed in rat lung membranes following preincubation with cholesteryl hemisuccinate. beta-Adrenergic receptor number increased by 50% without a change in antagonist affinity. However, beta adrenergic receptor affinity for isoproterenol increased 2-fold as a result of an increase in the affinity of the isoproterenol high-affinity binding site. The increase in agonist affinity did not potentiate hormone-stimulated adenylate cyclase activity, which decreased 3-fold following cholesterol incorporation. However, the ratio of isoproterenol to GTP-stimulated activity was unchanged with cholesterol. Stimulation distal to the receptor by GTP, NaF, GppNHp, Mn2+ and forskolin also demonstrated 50-80% reduced enzyme activity following cholesterol incorporation. These data suggest that membrane cholesterol incorporation decreases catalytic unit activity without affecting transduction of the hormone signal. PMID- 2988645 TI - Selective effects of CAPP1-calmodulin on its target proteins. AB - Occupancy of one of the two phenothiazine-binding sites on calmodulin does not significantly decrease the affinity of calmodulin for its target proteins; however, it does affect the ability of calmodulin to activate some enzymes. Previously we demonstrated that a covalent adduct of calmodulin with one molecule of phenothiazine (CAPP1-calmodulin) is an antagonist for the calmodulin-dependent enzymes, cAMP phosphodiesterase and myosin kinase, and a partial agonist for calcineurin. We now show that CAPP1-calmodulin is a full agonist for glycogen synthase kinase and phosphorylase kinase. Unlike phenothiazines, CAPP1-calmodulin is specific for calmodulin-regulated proteins; it has no effect on protein kinase C. With the exception of phosphorylase kinase, occupancy of two phenothiazine binding sites completely eliminates the ability of calmodulin to activate these proteins. Thus, the study of the interaction of CAPP1-calmodulin with calmodulin target proteins demonstrates that calmodulin interacts differently with different proteins. This is confirmed by studies of the effect of calmodulin fragments, 1 77 and 78-148, on calmodulin-regulated enzymes. PMID- 2988646 TI - [Changes in the properties of adenylate cyclase from guinea pig lungs in tuberculosis]. AB - Changes in the properties of adenylate cyclase from the lungs of tuberculotic guinea pigs were revealed. The number of beta-adrenergic receptors in the lungs was found to be reduced by 30% at the second and by 70% at the third stage of the disease. The degree and the value of Ka for adenylate cyclase activation by isoproterenol remained thereby unchanged. The basal activity of adenylate cyclase was increased by 20% against the control level at the second stage and decreased by 20% at the third stage of the disease. At these periods, the stimulating effects of guanylyl imidodiphosphate, NaF and forskolin on lung adenylate cyclase were diminished. The experimental results point to the significant role of the enzymes of cAMP metabolism and reflect the course of the tuberculosis process in experimental animals. PMID- 2988647 TI - [Factors reducing the superoxide dismutase activity of the liver of rats with Pliss lymphosarcoma]. AB - The procedures for isolation and purification of Cu/Zn superoxide dismutase (SOD) from small amounts of rat liver at different stages of Pliss lymphosarcoma growth were developed. Two stages of tumour growth were distinguished. At the first stage (4-5th day after reinoculation) the intensity of optical (680 nm) and EPR spectra of SOD was decreased, on the average, to 30%, while at the second stage- by 40% at 680 nm and by 32%, at 260 nm. The intensity of the EPR spectra was also diminished by 40% as compared to the control. It was assumed that at initial stages of tumour growth the decrease of the SOD activity is mainly due to the reduction of enzyme Cu, while at the second stage--to the decrease of the protein content. In all probability, the observed changes are induced by the activation of lipid peroxidation involving active O2 species. The second stage is associated with the superoxide-induced impairment of the function of nucleic acids involved in protein biosynthesis. PMID- 2988648 TI - [Peroxidase activity of catalase with respect to aromatic amines]. AB - The catalase dissociation into subunits has been studied at pH less than 3.5 and greater than 11.0. This process is characterized by pseudo-first order rate constants, depending on the initial concentrations of the enzyme and H+. At pH 2.85, the steady-state kinetics of five aromatic amines oxidation by catalase monomers has been studied for orthodianisidine (o-DA), 3,5,3',5' tetramethylbenzidine (TMB), ortho- and para-phenylene diamine (p-PDA) and 5 aminosalycilic acid. The optimal substrates for catalase in acidic solutions are o-DA, TMB and p-PDA. A comparison has been carried out for the catalase peroxidative activity, and the catalytic characteristics of horseradish peroxidase in the oxidation of the same substrate. The mechanisms of peroxidatic amines oxidation by catalase and horseradish peroxidase are discussed. PMID- 2988649 TI - [Kinetics of the interaction of methylene diphosphonic acid and inorganic pyrophosphate with DNA-dependent RNA-polymerase from calf thymus]. AB - The kinetics of interaction of PPi and its diphosphonic analog, methylenediphosphonic acid (MDPA), with nucleoside triphosphates, DNA and Mg2+ binding sites of DNA-dependent RNA polymerase II from calf thymus was investigated. The values of apparent Km in the NTP polymerization reaction for ATP and CTP equal to 2.7 X 10(-4) and 1.8 X 10(-4) M, respectively, were determined. It was shown that MDPA and PPi competitively inhibited the RNA polymerase reaction with respect to nucleoside triphosphate. The inhibition constants (Ki) of ATP and CTP incorporation for MDPA were 2.2 X 10(-4) and 3.3 X 10(-4) M, respectively, while those of the nucleoside triphosphate incorporation for PPi were equal to 1.4 X 10(-4) and 2.0 X 10(-4) M, respectively. MDPA and PPi were incompetitive inhibitors of template (DNA) and Mn2+. A possible mechanism of inhibition of the RNA polymerase reaction by MDPA is proposed. PMID- 2988651 TI - [Cytochrome c-induced acceleration of the reaction of muscle actin polymerization]. AB - The reaction of rabbit skeletal muscle actin polymerization initiated by the addition of neutral salts is accelerated in the presence of mitochondrial cytochrome c. The observed effect is specific, since the addition of serum albumin does not change the initial velocity of this process. The dependence of the rate of actin polymerization and exogenous cytochrome c concentration correlates within the molar protein ratios from 43:1 to 9:1, respectively. The increase in the polymerization rate occurs immediately after addition of cytochrome c to the reaction mixture; however, the maximal effect is observed only after 5 min coincubation of the proteins. The ability of cytochrome c to stimulate this process is abolished at alkaline values of pH (8.5), which points to the significant role of the molecule positive charge which, in all probability, serves as a primer of the muscle actin polymerization reaction. PMID- 2988650 TI - [ADP-ribosylation of histones and NAD-pyrophosphorylase activity of chicken liver nuclei during induction of DNA damage]. AB - The rate of [14C]NAD incorporation into chicken liver nuclear histones was studied under conditions of DNA damage by N-methyl-N-nitrosourea and pancreatic DNAase I. With an increase in N-methyl-N-nitrosourea concentration from 8.5 X 10( 2) to 34.0 X 10(-2) mM, the ADP ribosylation of histones increases by 20% as compared to the control. In DNAase I-treated nuclei, the binding by histones of [14C]NAD sharply increases, reaching its maximum (18.3 X 10(-8) mM) at 30% cleavage of DNA. When 50% of DNA was cleaved, the rate of [14C]NAD incorporation into the histones was 8.0 X 10(-8) mM as compared to 6.1 X 10(-8) mM/mg protein in control samples. The poly(ADPR)polymerase activity was increased in both cases. It was shown that the NAD-pyrophosphorylase activity in chicken liver nuclei treated with N-methyl-N-nitrosourea does not differ from the control one, while in DNAase I-treated nuclei the maximum of the NAD-pyrophosphorylase activity was achieved, as well as the maximum of [14]NAD incorporation into the histones within the range of DNA damage of 25-35%, being equal to 37 X 10(-8) mM NAD/min/mg protein as compared to 26.0 X 10(-8) mM/min/mg protein in the control. At different degrees of DNA damage, the average length of the poly-ADP-ribose chain did not practically alter, thus suggesting the increase in the number of polymer binding sites in the histones. PMID- 2988652 TI - [Restriction map of maxicircle kinetoplastic DNA of Leishmania gymnodactyli]. AB - Kinetoplast DNA (kDNA) of Leishmania gymnodactyli, as in other Trypanosoma species, is a complicated associate consisting of mini- and maxicircular molecules. Minicircular DNA are presented by the homogeneous in size and heterogeneous in base sequence population. Their size determined by the agarose electrophoretic mobility in gel is 920 pairs of nucleotides. Maxicircular molecules of DNA are homogeneous and contains about 38 thou pairs of nucleotides. The restriction map of maxicircular kDNA molecule of L. gymnodactyli has been made up on the basis of the cleavage data of the whole associate kDNA by restrictases. PMID- 2988653 TI - 5'-Nucleotidase activities in human fetus. AB - Specific 5'-nucleotidase (5'-N) activity in 17 human fetuses was systemically studied in order to get basic values of the enzyme activity. A wide spectrum of 5'-N activity among the organs was revealed. Very strong activity was found in placenta (42.62 +/- 19.24 nmol adenosine/mg protein/min), skeletal muscle (31.80 +/- 11.10) and skin (27.31 +/- 13.00); strong activity in liver (18.96 +/- 7.60), pituitary (15.06 +/- 6.13) and thyroid (12.69 +/- 4.47); moderate activity in lung (8.43 +/- 3.68), pancreas (8.20 +/- 2.26), small intestine (8.07 +/- 2.28), lymph node (7.26 +/- 2.22), thymus (7.06 +/- 2.52), spinal cord (5.86 +/- 1.96) and spleen (5.65 +/- 2.52), and weak activity in testis (4.02 +/- 0.79), heart (3.95 +/- 1.22), adrenal (3.46 +/- 1.68), kidney (3.16 +/- 0.83), ovary (3.16 +/- 0.72), aorta (2.98 +/- 1.20), cerebellar hemisphere (2.28 +/- 1.09) and frontal lobe of the cerebrum (1.49 +/- 0.54). These data are the first to be reported on specific 5'-N activity in the human fetus. The significance of the wide range of 5'-N activity among the organs, and future aspects on the study of 5'-N activity with respect to cell differentiation, maturation, development, aging or malignant transformation were discussed. PMID- 2988654 TI - Postnatal changes in hepatic microsomal enzyme activities in the puppy. AB - The in vitro activities of glucose-6-phosphatase (G6P), UDP-glucuronyl transferase (GT) and P-450 were measured in liver homogenate and/or microsomal suspensions from puppies, 0-42 days of age (n = 26), and adult dogs (n = 3). For each of these enzymes, an age-related increase in the in vitro activity was observed, with the lowest value detected at birth. By the 28th-42nd day of postnatal life, P-450-specific activity was 350 and 85%, G6P 225 and 188%, p nitrophenol GT 430 and 105% and bilirubin GT 317 and 123% of that seen in 0-hour old puppies and adults dogs, respectively. The age-related changes in G6P and GT were observed when native enzymes or enzymes activated by deoxycholate or UDP-N acetylglucosamine (GT) were used. However, the ratio between activated and native p-nitrophenol GT activity decreased as a function of age, and UDP-N acetylglucosamine failed to activate bilirubin GT in puppies of 0-42 days of age. Total liver protein also increased with age, and hepatic water content was significantly higher in 0- to 42-day-old puppies (76.3%) than in adult dogs (71.4%). Thus, differences between puppies and adult animals were not the same when protein content or enzyme activities were expressed per unit of wet or dried liver weight. Phenobarbital, injected intraperitoneally at 15 mg/kg/day for 6 consecutive days to 8- to 13-day-old puppies (n = 3), produced induction of P-450 (235% of age-matched controls) and bilirubin GT activity (160%), diminished G6P activity (81%), and failed to modify p-nitrophenol GT activity (102%). These studies indicate that, in the puppy, (1) the in vitro activities of P-450, G6P and GT are immature at birth and develop during postnatal life; (2) as in other species, bilirubin and p-nitrophenol may be conjugated in the dog liver by two functionally distinct GT. PMID- 2988655 TI - Erythrocyte membrane sodium-potassium and magnesium ATPase in primary affective disorder. AB - Erythrocyte membrane Mg2+ ATPase and Na+-K+ ATPase were measured in patients with affective disorder, their well relatives, and normal controls during euthymic moods. On the average, the Mg2+ ATPase activity was high in subjects belonging to affective disorder families. However, the difference between normal and affective disordered individuals was not statistically significant. Only the well individuals from affective disorder pedigrees as a group had significantly higher than normal Mg2+ ATPase activity (p less than 0.05). The Na+-K+ ATPase activity was similar for all the groups, including normal, bipolar manic-depressive (with or without lithium), unipolar depressive, and well individuals. Lithium treatment did not seem to have any effect on Mg2+ ATPase. Even though the values of Na+-K+ ATPase in the lithium-treated group were high, it is not certain that this was due to lithium per se. PMID- 2988656 TI - The relationships of historically defined subtypes of depression to ACTH and cortisol levels in depression: preliminary study. AB - A family history of depression (but no alcoholism), a history of bipolarity, and a history of nonsuppressor status on the Dexamethasone Suppression Test (DST) have all been positively associated with each other in previous studies. We divided depressives into three mutually exclusive groups, using the three historical parameters described above. Group A included those who were nonsuppressors at index. Group B included normal suppressors at index who met one of the following three criteria: (1) past history of a nonsuppressing DST, (2) past history of a mania, and (3) family history of depression (but no alcoholism). The remaining suppressors at index made up Group C. We found that Groups A and B show a phase advance (an earlier nadir) in the predexamethasone circadian curve for cortisol. Adrenocorticotrophic hormone (ACTH) varies in part (but not solely) with cortisol and may separate the groups. PMID- 2988657 TI - Regulation of infant and developing rat testicular gonadotropin and prolactin receptors and steroidogenesis by treatments with human chorionic gonadotropin, gonadotropin-releasing hormone analogs, bromocriptine, prolactin, and estrogen. AB - Infant (5-day-old) male rats were treated with hormonal regimens to alter their exposure to gonadotropins, prolactin (Prl), and estrogen, and the response of testicular endocrine functions was measured. Human chorionic gonadotropin (hCG) or a potent gonadotropin-releasing hormone agonist analog (GnRH-A) resulted in a short-lived decrease of testicular receptors (R) for luteinizing hormone (LH), but no deleterious effects were found on testicular capacity to produce testosterone (T), which is a typical response of the adult testis. Only GnRH-A, through probable direct testicular action, induced a relative blockade of C21 steroid side-chain cleavage that was observed in vitro upon hCG stimulation. Human chorionic gonadotropin treatment, but not GnRH-A treatment, increased testicular Prl-R. GnRH antagonist analog (GnRH-Ant) treatment did not affect testicular LH-R, but decreased Prl-R and testicular T production. Decrease of serum Prl by bromocriptine had no effect on testicular LH-R or Prl-R, but slightly decreased T production in vitro. Ovine Prl increased binding sites for LH/hCG. The postnatal rats were insensitive to negative effects of diethylstilbestrol when monitored by testis weight, T, and LH-R. In conclusion, the responses to changes in the hormonal environment differed greatly between infant and adult testes. Mainly positive effects of elevated gonadotropin and Prl levels were seen on infant rat Leydig cell functions. Likewise, decreased tropic hormone levels, and exposure to estrogen, were ineffective in bringing about the inhibitory actions seen in the adult. PMID- 2988658 TI - Studies on the mechanism of follicle-stimulating hormone-induced desensitization of Sertoli cell adenylyl cyclase in vitro. AB - When Sertoli cells were cultured in the presence of follicle-stimulating hormone (FSH), a time-and concentration-dependent desensitization of FSH-responsive adenylyl cyclase (AC) was observed. Maximal desensitization (80%) was attained after 6-9 h of incubation with FSH (10 micrograms/ml; NIH-FSH-S12). During 24 h of incubation the concentration of FSH causing a half-maximal desensitization was about 100 ng/ml. Removal of the hormone from the culture medium was associated with a gradual reappearance of the FSH response. Follicle-stimulating hormone induced desensitization of Sertoli cell AC was specific for homologous hormone, since AC activation by isoproterenol was unaffected. Furthermore, AC activity of control and FSH-desensitized cells was equally activated by GTP and fluoride, showing that the interaction of the guanyl nucleotide regulatory (N) component with the catalytic subunit is not affected during FSH-induced desensitization. A loss in specific FSH binding was detected after 9 and 24 h of exposure to FSH, but not at shorter times of incubation. Desensitization of Sertoli cell AC to both FSH and isoproterenol stimulation could also be achieved by dibutyryl cyclic AMP (dbcAMP); however, a 30-40% desensitization required a high nucleotide concentration (1 mM) and a long incubation time (24 h). These results show that desensitization of Sertoli cell AC by FSH is associated with normal function of the N component, and precedes any significant loss in specific FSH binding sites. Furthermore, exogenous addition of dbcAMP (1 mM) did not cause the same effects on Sertoli cell AC as did FSH. PMID- 2988659 TI - The shear strength between bone and porous ceramic root implants in the guinea pig incisor socket. AB - Porous alumina and apatite implants have been placed in 67 guinea pig incisor tooth sockets after tooth extraction under anaesthetic. Bone readily grew into the spaces of the implant, and reduced the fibrous connective tissue between implant and bone. 'Pushout' tests have monitored the shear strength of the interlock between bone and implant. Despite considerable variation in the interlock strength, apatite produced the strongest bone interlock. The interlock with alumina tended to be more fibrous. The relative merits of fibrous and bone interlocks in immediate root implants are considered for the augmentation of the alveolar ridge and prosthetic treatment. PMID- 2988660 TI - Bone microstructure by collagenase etching. AB - A novel technique has been developed for microstructural studies of bone. The spatial organizations of the mineral and collagen fibres in bone have been a matter of discussion for some time, with numerous diverse observations arising from various preparative techniques. In this latest investigation details of the mineral structure are clearly revealed in the SEM by treating a cut and polished surface of bone with collagenase to remove the major organic component. This new procedure has minimal effect on the mineral and hence reveals microstructural detail which is far closer to that in vivo than in previous investigations. This paper concentrates on two aspects of the studies, namely the detailed morphology of the mineral component and the arrangement of the collagen fibres in the osteons of compact bone. Firstly, the mineral component is revealed as comprising 'crystallites' (approximately 20 nm diam.) which aggregate to form larger contiguous 'spheroidal particles' (approximately 100 nm diam.), which in turn form 'granules' (approximately 500 nm diam.). Secondly, the regions from which collagen fibres have been removed are clearly revealed, showing that within an osteon, alternating lamellae have collagen fibres oriented approximately parallel to and circumferential to the Haversian canal respectively. PMID- 2988661 TI - [Analysis of disordered muscle contractile function by assessments of the degree of potentiation of evoked mechanical response of the muscle]. AB - A study was made of the evoked mechanical response of the adductor under supramaximal stimulation of the ulnar nerve in 24 normal test subjects, 10 patients with the involvement of motoneurons and their axons, 30 patients with chronic derangement of the neuromuscular transmission (myasthenia) and 8 patients with hypothyrosis. A reverse dependence was found of the Pt/Po ratio on the magnitude of post-tetanic potentiation in all the examined. The capacity of the muscle to potentiate the tension in response to the double stimulus and during the post-tetanic periods inversely related to the initial amplitude of the evoked mechanical response of the muscle and the time of the muscle contraction. It is assumed that the universal mechanism of the regulation of muscle contractile function in health and disease underlies the regularities described. PMID- 2988662 TI - [Interrelation of immune and mediator lymphocyte receptors in mice]. AB - A study was made of the action of a specific muscarinic antagonist 3H quinuclidinyl benzylate on the immune rosette formation in BALB/c mice. It was shown that treatment of mouse spleen lymphocytes by 3H-QNB at a concentration of 10(-9) M-10(-14) M brought about rosette formation inhibition. The process was dose-dependent. Atropine reversed the action of 3H-QNB. PMID- 2988663 TI - [Induction of chromosome aberrations by the T antigen of the SV40 virus introduced into the cells using liposomes]. AB - A purified SV40 T antigen introduced into hamster cells by means of liposomes accumulated in the nuclei within 10 h and persisted there further as long as 10 to 12 h. Within the first day after cell treatment with T antigen numerous chromosome aberrations including breaks, translocations and gaps were observed in the cells. The number of aberrations slightly reduced by the 2nd day followed by restoration of the normal cell karyotype by the 5th day. Removal of T antigen incorporated into liposomes by a specific immunosorbent or heat inactivation of T antigen abolished the clastogenic effect. It is suggested that induction of chromosome aberrations might activate sell protooncogenes and thus serve a genetic basis of tumor progression. PMID- 2988664 TI - Regulation of receptors and digestive activity toward synthesized formyl chemotactic peptide in human polymorphonuclear leukocytes. AB - A receptor binding and digestive activity of human polymorphonuclear leukocytes (PMNs) toward formyl-methionyl-leucyl-[3H]phenylalanine (3H-FMLP) was examined with the following results: Up- and down-regulation and recovery of 3H-FMLP binding activity were demonstrated. Both intact PMN and a lysate prepared from them cleaved the carboxyl terminal amino acid (phenylalanine) of 3H-FMLP. The digestive activity decreased as the receptor binding was inhibited by n ethylmaleimide and 4-chloromercuribenzoate. Little digestive activity was found in the supernatant from PMN stimulated by FMLP. The released phenylalanine was found in the pellet and supernatant of PMNs. Digestive activity with cathepsin A like characteristics was found in the lysate of PMN. These observations suggest that FMLP is internalized in lysosomes in a receptor-mediated manner and cleaved by the cathepsin A-like enzyme, the free phenylalanine is released extracellularly, and a part of the dissociated receptors with FMLP may return to the surface or to an intracellular receptor pool. Another finding was that the digestive activity of the lysate of cord blood granulocytes was decreased compared with that of adult blood granulocytes. This decrease may explain in part the impaired chemotaxis of cord blood granulocytes. PMID- 2988665 TI - Preleukemic state of adult T cell leukemia: abnormal T lymphocytosis induced by human adult T cell leukemia-lymphoma virus. AB - We report the clinical, hematologic, and immunologic features of 18 preleukemic adult T cell leukemia (pre-ATL) cases with abnormal T lymphocytosis induced by human adult T cell leukemia-lymphoma virus (HTLV/ATLV). The patients were from the Nagasaki district, which is one of the most endemic areas of ATL in Japan. Pre-ATL is a subclinical T cell abnormality differing from ATL. It is characterized by an insidious onset and appearance of abnormal T lymphocytes (10% to 40%) in the peripheral blood without clinical symptoms except for a few cases transiently presenting fever, skin eruptions, and slight lymphadenopathies. Most abnormal T lymphocytes were small and mature with incised or lobulated nuclei and formed E rosettes with sheep RBCs. Virologic and biomolecular analysis revealed that all cases were infected with HTLV, and proviral DNA was integrated in host lymphocytes from 12 of the 14 cases examined. Furthermore, the lymphocyte populations, including abnormal T lymphocytes, were monoclonal with respect to the site of the provirus integration. Abnormal T lymphocytosis persisted from one to more than seven years in six cases, three of which developed ATL after a one- to five-year pre-ATL stage, whereas abnormal T lymphocytes spontaneously decreased in the other seven patients. However, HTLV-infected monoclonal lymphocytes were detected in four cases examined, even after most of the abnormal T lymphocytes had disappeared. Moreover, the same clonally provirus-integrated lymphocytes persisted in two of four cases not only during the course of abnormal lymphocytosis, but also in the subsequent almost-normal blood. These results indicate that the majority of the cases were in a pre-ATL state with a potential to develop ATL. PMID- 2988666 TI - Neutrophils adherent to a nonphagocytosable surface (glomerular basement membrane) produce oxidants only at the site of attachment. AB - Adherence of neutrophils to glomerular basement membrane containing immunoglobulin G aggregates was accompanied by a marked increase in oxygen uptake (eightfold). Very little of the O2 consumed was recovered as superoxide, measured by cytochrome c reduction, or as H2O2, measured with horseradish peroxidase and scopoletin. When neutrophils were incubated with the basement membrane preparation in the presence of cerium chloride to detect H2O2, electron micrographs showed cerium perhydroxide deposits in the contact area between the cells and the basement membrane, but not on the remainder of the cell surface. The results imply that superoxide is produced only where the plasma membrane is in contact with the basement membrane matrix, and that it mostly breaks down to H2O2 or undergoes other reactions at this site. The longer lifetime of H2O2 compared with that of superoxide allows some of the H2O2 produced to be detected in the medium. The results also suggest that the area of contact between the neutrophil and surfaces such as basement membrane is inaccessible to proteins in the medium, eg, cytochrome c. Circulating scavengers such as superoxide dismutase or catalase, or proteolytic inhibitors, may therefore be unable to control events occurring at this site. PMID- 2988667 TI - Platelet-derived growth factor promotes human peripheral monocyte activation. AB - Like in the polymorphonuclear leukocyte (PMN), the platelet-derived growth factor (PDGF) purified to homogeneity is capable of inducing monocyte activation responses as evaluated by generation of superoxide anion (O-.2) from membrane associated oxidase system, release of granule enzymes, and enhanced cell adherence and cell aggregation. Superoxide anion release was maximized at 10 ng/mL PDGF and was comparable to that induced by 10(-7) mol/L formyl-methionyl leucyl-phenylalanine. The potency of PDGF to induce this response in monocytes was of the same magnitude as that observed in PMNs. Similarly, lysozyme release and monocyte adherence were also increased in a dose-dependent manner and achieved maximal responses at 40 ng/mL concentration of PDGF. The PDGF concentration required to achieve maximal monocyte aggregation was two-fold (60 ng/mL) of that found for PMNs. In contrast to PMNs, a positive correlation (gamma = .93; P less than .01) was observed between the increases of PDGF concentration and beta-glucuronidase release. These findings indicate that PDGF can induce the full sequence of cell activation events in human monocytes similar to human PMNs. PMID- 2988669 TI - Hydrops fetalis due to an unusual form of Hb H disease. AB - The occurrence of Hb H hydrops fetalis is reported for the first time. The mother has zeta-alpha thalassemia 1 (zeta zeta alpha alpha/----) and the father has non deletion alpha thalassemia [zeta zeta alpha alpha/zeta zeta (alpha alpha)T]. The complete deletion of the zeta alpha cluster on one chromosome was confirmed by quantitation of alpha and zeta gene numbers, the normal alpha and zeta gene patterns arising from the remaining normal chromosome, and the decreased alpha/beta globin chain ratio of 0.57. The non-deletion alpha thalassemia defect could only be identified by the imbalanced alpha/beta globin chain ratio of 0.65 in the presence of normal gene numbers and patterns. The newborn was markedly anemic, unlike those with classical Hb H disease, because the non-deletion alpha thalassemia defect is more severe than alpha thalassemia 2. The decreased zeta genes during fetal life might have additional deleterious effects. In this family, the distinct BamHI restriction fragment length polymorphism in the hypervariable region of the zeta genes may be used for future prenatal diagnosis. PMID- 2988668 TI - Zinc-induced platelet aggregation is mediated by the fibrinogen receptor and is not accompanied by release or by thromboxane synthesis. AB - We demonstrate that zinc (0.1 to 0.3 mmol/L) induces aggregation of washed platelet suspensions. Higher concentrations (1 to 3 mmol/L) of zinc were needed to aggregate platelets in platelet-rich plasma obtained from blood anticoagulated with low-molecular-weight heparin, probably due to the binding of zinc to the plasma proteins. Zinc-induced aggregation of normal washed platelets required added fibrinogen and no aggregation occurred with thrombasthenic platelets or with normal platelets pretreated with a monoclonal antibody (10E5) that blocks the platelet fibrinogen receptor. These data indicate that the platelet membrane fibrinogen receptor-glycoproteins IIb and IIIa mediate the effect of zinc. Zinc induced aggregation was blocked by the agent TMB-8, which interferes with the internal calcium flux, and by prostacyclin, which elevates platelet cyclic adenosine monophosphate levels. Zinc-induced aggregation was not accompanied by thromboxane synthesis or by the secretion of dense-body serotonin and was not affected by preexposure of platelets to acetylsalicylic acid. Experiments with creatine phosphate/creatine phosphokinase showed that the zinc effect on platelets was independent of extracellular adenosine diphosphate (ADP). Zinc had an additive effect when platelet aggregation was stimulated with subthreshhold concentrations of collagen or ADP. Together with the known effects of nutritional zinc on in vivo bleeding, on platelet aggregation, and on lipid metabolism, the results suggest that zinc may have an important bearing on normal hemostasis, thrombosis, and atherosclerosis. PMID- 2988670 TI - Variable breakpoints on the Philadelphia chromosome in chronic myelogenous leukemia. AB - The abl oncogene is translocated from chromosome 9 to 22 in the creation of the Philadelphia (Ph1) chromosome. This article describes new translocation breakpoints identified in two patients with chronic myelogenous leukemia using Southern blotting and cloned human DNA probes from chromosome 9. The translocation breakpoints on chromosome 9 in both of these patients lie closer to the human cellular abl (c-abl) gene, and the chromosome 22 breakpoints are distributed more widely than previously reported. These data help to define more clearly the chromosomal span of the breakpoints and indicate that some translocations include very little chromosome 9 sequence located 5' to the c-abl gene. PMID- 2988672 TI - ATP produces vasodilation via P1 purinoceptors and vasoconstriction via P2 purinoceptors in the isolated rabbit central ear artery. AB - P1 and P2 purinoceptors mediating mechanical responses in isolated rabbit ear artery were studied by comparing responses to adenosine triphosphate (ATP), alpha, beta-methylene ATP, and adenosine, both when endothelial cells were intact and when they had been removed by mechanical rubbing (as confirmed by histochemical staining and near abolition of relaxation to acetylcholine). alpha, beta-Methylene ATP and ATP (but not adenosine or acetylcholine) contracted preparations at resting tone. alpha, beta-Methylene ATP was significantly more potent as a contractile agent than ATP. Neither was significantly affected by removal of the endothelium. Acetylcholine, ATP, and adenosine relaxed arteries whose tone had been raised by 10(-6) M histamine. Removal of the endothelium virtually abolished relaxations to acetylcholine and significantly decreased those to adenosine and ATP. All relaxations to adenosine and ATP were significantly antagonised by 8-phenyltheophylline, a potent P1 purinoceptor antagonist. alpha, beta-Methylene ATP further contracted the high-tone preparation and was again unaffected by removal of the endothelium. These results confirm that endothelial cells can mediate vasodilation. They also show that adenosine (and ATP) can elicit vasodilation via P1 purinoceptors both on the endothelial cells and on the smooth muscle of the isolated rabbit central ear artery. P2 purinoceptors, however, appear to be located on the smooth muscle only and mediate vasoconstriction. PMID- 2988671 TI - Cell-lineage antigens of the stem cell-megakaryocyte-platelet lineage are associated with the platelet IIb-IIIa glycoprotein complex. AB - The stem cell-platelet lineage is uniquely defined by platelet cell-lineage antigens. These antigens are present on all stem cells measured by the spleen colony assay and become restricted to the platelet cell lineage as differentiation proceeds. In this study, anti-platelet serum (APS) has been used to identify cells in the bone marrow that express platelet cell-lineage antigens and to identify platelet cell surface molecules expressing these antigens. Anti platelet IgG extensively absorbed with brain, thymus, and peritoneal cells bound selectively to stem cells, megakaryocyte progenitor cells (Mk-CFC), and megakaryocytes in CBA mouse bone marrow and to blood platelets. No other hemopoietic cell type, tissue, cell line, or tumor cell bound significant amounts of antibody against platelet cell-lineage antigens as determined by ability to absorb the anti-stem cell activity in APS. Studies with lactoperoxidase-labeled platelets showed that two major iodinated proteins of Mr = 114,000 and 138,000 were immunoprecipitated with APS and with antiserum that had been extensively absorbed. These proteins correspond to the platelet IIb-IIIa glycoprotein complex, which is known to express receptors for collagen and fibrinogen, molecules known to influence hemopoietic cell proliferation and tumor cell growth. A panel of six monoclonal antibodies against human IIb-IIIa inhibited spleen colony formation by 17% to 100%, J15 and A5.15 also being cytotoxic for granulocyte-macrophage progenitor cells and Mk-CFC. Other platelet monoclonal antibodies did not inhibit spleen colony formation. Although APS inhibited fibrinogen binding to platelets and platelet aggregation, these activities were greatly reduced with absorbed antiserum. Furthermore, fibrinogen treatment of bone marrow did not block the anti-stem cell activity in APS. Thus the evidence is consistent with expression of platelet cell-lineage antigens on the platelet IIb-IIIa glycoprotein complex at a site removed from the fibrinogen binding site. PMID- 2988673 TI - Thymosin beta 4 induced phenotypic changes in Molt-4 leukemic cell line. AB - Thymosin beta 4 was tested for its ability to induce phenotypic changes in the human T-cell line Molt-4. Cells were cultured with nanogram concentrations of thymosin beta 4 for up to 16 days and were analyzed with a panel of monoclonal antibodies, sheep erythrocyte rosetting, peanut agglutinin binding (PNA) and an antibody to the enzyme, terminal deoxynucleotidyl transferase (TdT). Thymosin beta 4 induced Molt-4 cells to reduce the expression of a T-cell lineage specific antigen, with preferential expression on T blast-cells, detected by WT 1 monoclonal antibody. Thymosin beta 4 also induced an increase in sheep erythrocyte rosettes and PNA binding as well as an increased expression of OKT 11 A and OKT 8 in Molt-4 cells. TdT was found to be unchanged, however. Analysis of thymosin beta 4-treated cells with other monoclonal antibodies (OKT 3, OKT 6, OKT 9) showed no change when compared to controls. These results showed that thymosin beta 4 is capable of inducing phenotypic changes in Molt-4 cells. Such changes may represent a differentiation process of these cells through the early stages of the maturation process of thymus-dependent lymphocytes, albeit not to the stage of mature T cells. PMID- 2988675 TI - [Development of aggressive cystosarcoma phyllodes after conservative treatment of breast adenocarcinoma]. AB - The case report is that of patient who underwent tumorectomy plus X-ray therapy for mammary invasive ductal carcinoma. Eight years later a large cystosarcoma phyllodes, developed and quickly recurred after conservative surgery. Reviewing previous mammograms, the authors discovered that at the time of irradiation, there was a small lump which had the appearance of fibroadenoma. It may be that heavy X-ray therapy on a benign epithelial and stromal tumour of the breast, played a major role in the later development of cystosarcoma phyllodes. PMID- 2988676 TI - Intrapulmonary shunt is not increased by 100% oxygen ventilation in acute respiratory failure. AB - Increase of shunt has been demonstrated during short periods of pure oxygen breathing, mostly in patients with mild acute respiratory failure (ARF). Twenty patients with a large range of venous admixture (12 to 63%) were studied when FIO2 was increased from maintenance to one. Intrapulmonary shunt was measured with both the conventional oxygen method [QS/QT (O2)] and the multiple inert gas elimination technique [QS/QT (IG)]. Mean venous admixture decreased from 29 to 24% when FIO2 was increased and QS/QT (IG) remained unaltered. The pattern of blood flow distribution remained similar in both conditions, even in the eleven patients with ARF secondary to bacterial pneumonia and who had a low V/Q mode, highly liable to collapse with 100% oxygen. As no increase in shunt was documented in these patients, reluctance to measure shunt during 100% oxygen breathing should be re-evaluated. PMID- 2988677 TI - Enhancement of glomerular permeability to anionic ferritin induced by kidney perfusion with collagenase. AB - The role of collagen in ultrafiltration properties of the glomerular basement membrane (GBM) was tested after a single administration of bacterial collagenase, using native ferritin as a tracer which does not pass through the GBM under physiological conditions. Experiments were performed both in situ and with isolated kidneys. Increased permeability to ferritin occurs 6 hr following enzyme perfusion and becomes patent after 30 hr, numerous tracer molecules appearing in urinary space, without any readily observable changes either in distribution of fixed negative charges (as revealed by colloidal iron and polyethyleneimine) or in structural organization of the glomerulus. Selective permeability of the GBM is progressively restored so that ferritin is almost confined to capillary lumen one month after enzyme injection. We conclude that collagen plays an important part in restricting plasma protein filtration. PMID- 2988674 TI - [Demonstration of calmodulin in cultured human malignant melanocytes by indirect immunofluorescence]. AB - Using calmodulin antibody, evidence for the presence of calmodulin, a calcium binding protein modulator of numerous enzymes was shown by indirect immunofluorescence in 9 cell lines of cultured human malignant melanocytes. Calmodulin was localized in cytoplasm and in higher concentration in the perinuclear region. These observations agree with the distribution of calmodulin binding proteins in cytoplasm and membranes. PMID- 2988678 TI - Transforming growth factors--an overview. AB - Transforming Growth Factors (TGFs) are a sub-group of a larger family of protein hormones. Two major types of TGF are currently known, alpha-TGF and beta-TGF. Biologically their most important property is to act synergistically to induce anchorage-independent growth of target cells otherwise incapable of such growth. Since this growth parameter is well correlated with tumorigenicity in vivo, these factors may play a role in cancer development. Biochemically both alpha-TGF and beta-TGF have been well characterised in some types of cells and tissues. Their role in normal and neoplastic growth is actively studied. PMID- 2988679 TI - Actions of adenine dinucleotides in the guinea-pig taenia coli: NAD acts indirectly on P1-purinoceptors; NADP acts like a P2-purinoceptor agonist. AB - The actions of the adenine dinucleotides beta-nicotinamide adenine dinucleotide (NAD) and beta-nicotinamide adenine dinucleotide phosphate (NADP) were examined on the carbachol-contracted taenia coli of the guinea-pig. Both were capable of inducing full relaxations in a concentration-dependent manner; NADP was 21.4 times more effective than NAD at EC50; the threshold for NADP was approximately 0.1 microM and for NAD approximately 1.0 microM. The P1-purinoceptor antagonist, 8-phenyltheophylline (10 microM), produced a large parallel rightward shift in the NAD concentration-response curve; in contrast it produced a small parallel leftward shift in the NADP concentration-response curve. Dipyridamole (0.2 microM), a purine nucleoside uptake inhbitor, markedly potentiated responses to NAD and slightly potentiated NADP. 8-Phenyltheophylline antagonized the dipyridamole potentiation of both NAD and NADP. By use of high performance liquid chromatography it was shown that the action of NAD involves a breakdown to adenosine. Apamin, a K+ channel blocker, which antagonizes P2-purinoceptor activation in the intestine, abolished responses to NADP but not to NAD. The alpha-and beta-adrenoceptor antagonists, phentolamine (1 microM) and propranolol (1 microM), did not affect responses to NAD or NADP. Tetrodotoxin, a neurotoxin, did not abolish responses to either NAD or NADP. It is concluded that NAD acts largely indirectly as a P1-purinoceptor agonist following its breakdown to adenosine by ectoenzymes, while NADP acts in a similar manner to a P2 purinoceptor agonist. PMID- 2988680 TI - Baclofen activates voltage-dependent and 4-aminopyridine sensitive K+ conductance in guinea-pig hippocampal pyramidal cells maintained in vitro. AB - The ionic mechanism underlying the effect of (-)-baclofen in the hippocampus was investigated using guinea-pig brain slices. (-)-Baclofen either perfused or applied directly by microiontophoresis hyperpolarized the membrane and decreased the membrane input resistance of pyramidal cells in a dose-dependent manner. The value of the reversal potential for the baclofen-induced hyperpolarization, as estimated from the current-voltage relationships, was about -95mV. The reversal potential of the baclofen-induced hyperpolarization measured directly coincided with that for the post-burst hyperpolarization which is known to result from an activation of Ca2+-activated K+ conductance. The amplitude of the baclofen induced hyperpolarization was increased in low K+ (1.24 mM) medium whereas the hyperpolarization was decreased or abolished in high K+ (12.4 and 25 mM). Low Cl- (10.2 mM) medium had no noticeable effect on the baclofen-induced hyperpolarization. The effect of baclofen was antagonized by a low dose of 4 aminopyridine (5 X 10(-6) M) whereas it was unaffected by picrotoxin (2 X 10(-5) M). These results strongly suggest that the effect of baclofen is mediated by an increase in K+ conductance. PMID- 2988681 TI - Characterization of pre- and postsynaptic actions of (-)-baclofen in the guinea pig hippocampus in vitro. AB - The effects of (-)-baclofen on evoked potentials in the hippocampus were examined through intracellular recordings from guinea-pig brain slices. The evoked responses were recorded in two fibre connections within the hippocampus: the Schaffer collateral/commissural-CA1 pyramidal cell, and the mossy fibre-CA3 pyramidal cell. The Schaffer collateral/commissural-CA1 response was suppressed by (-)-baclofen in concentrations over 2 X 10(-5)M, whereas (+)-baclofen, an inactive isomer, in a concentration of 10(-4)M had no effect on the response. A compound action potential of Schaffer collateral/commissural axons was unaffected by (-)-baclofen even at 10(-4)M, a concentration that almost completely depressed the evoked response in the CA1 pyramidal cell. The mossy fibre-CA3 response was not inhibited by (-)-baclofen (10(-4)M). The depressant action of (-)-baclofen on the Schaffer collateral/commissural-CA1 response was unaffected by bicuculline (10(-4)M), whereas the direct membrane effects of (-)-baclofen were antagonized by bicuculline (10(-5)M). It is suggested that (-)-baclofen may modulate neuronal transmission through presynaptic recognition sites possibly by decreasing transmitter release from nerve terminals and also may directly regulate the endogenous neuronal excitability through an activation of the postsynaptic recognition sites. PMID- 2988682 TI - Modulatory activity of GABAB receptors on cholinergic tone in guinea-pig distal colon. AB - The effect of gamma-aminobutyric acid (GABA) administration was studied in both in vitro and in vivo preparations of the guinea-pig distal colon. In in vitro preparations GABA (10(-7) - 10(-3) M) elicited a dose-dependent relaxation; a decrease in the spontaneous contractions was sometimes observed. The effect of GABA was mimicked by (-)-baclofen, which gave a dose-response curve overlapping that of GABA, while (+)-baclofen was about one hundred times less potent. The relaxation responses induced by the above drugs were antagonized by 5 aminovaleric acid (5 X 10(-4) M), which did not affect adenosine-induced relaxation, but they were insensitive to bicuculline (10(-5) M) and picrotoxin (10(-5) M). Moreover, they were prevented by tetrodotoxin (6 X 10(-7) M). In hyoscine (10(-7) M)-pretreated preparations, GABA still evoked a small relaxation response (approx. 10% of the maximum) that was bicuculline-sensitive. Desensitization to GABA (10(-5) M) was observed. A specific cross-desensitization occurred between GABA (10(-5) M) and (-)-baclofen (10(-5) M). In in vivo preparations, GABA (10 mumol kg-1) and (-)-baclofen (5 mumol kg-1) produced a dose-related inhibition of basal tone, while (+)-baclofen (5 mumol kg-1) had much less effect (about 25%). A decrease in the spontaneous contractions was sometimes observed. The relaxant effect of GABA and (-)-baclofen persisted in guinea-pigs pretreated (1-2 min) with picrotoxin (1.6 mumol kg-1), whereas it was significantly reduced in animals injected 1 min beforehand with 5-aminovaleric acid (0.2 mmol). The maximal relaxant effect induced by GABA and (-)-baclofen did not differ from that of atropine (0.9 mumol kg-1) and after atropine administration GABA had no further inhibitory effect. Relaxation responses induced by GABA and (-)-baclofen still occurred after blockade of nicotinic receptors by hexamethonium (0.17 mmol kg-1), which itself caused an increase in the basal tone. When the tone was increased by topical application of physostigmine (40 micrograms), GABA and (-)-baclofen induced a greater relaxation than that obtained in basal conditions. It is concluded that GABA, both in vitro and in vivo administration, inhibits cholinergic tone in guinea-pig distal colon and that this effect is mediated mainly by activation of GABAB receptors. Further experiments are required to ascertain the possible physiological role of a GABA releasing neuronal system in the colon in vivo. PMID- 2988683 TI - Excitatory neurotensin receptors on the smooth muscle of the rat fundus: possible implications in gastric motility. AB - Picomolar concentrations of neurotensin caused concentration-dependent contractions of the longitudinal musculature of the fundus of the rat stomach. The EC50 of neurotensin was approximately 1.5 nM. On a molar basis neurotensin was about 5-10 times more potent than 5-hydroxytryptamine (5-HT) and approximately 80 times as active as acetylcholine in producing similar contractions. Studies with structurally related peptides indicated that whereas the carboxy terminal portion of neurotensin was essential for biological activity, a substantial part of its amino terminus end could be removed without affecting its potency. The EC50 for the neurotensin fragment 8-13 was identical to that of neurotensin, however its 1-8 or 1-11 fragments were completely inactive. Tetrodotoxin did not modify the potency of neurotensin or structurally related analogues suggesting that the neurotensin receptor is probably located on the smooth muscle membrane. In addition, the potency of neurotensin in contracting the fundus was not modified by pretreatment with atropine, methysergide or diphenhydramine. Fade to the contractile response of neurotensin was followed by the development of tachyphylaxis; desensitization was concentration-dependent and characterized by a shift in the agonist concentration response curve to the right and downwards. Desensitization with a priming concentration of neurotensin (approx. EC50) caused a substantial blockade of its excitability. There was cross-desensitization between neurotensin and the contractile activity of neurotensin 8-13 or xenopsin, but not with angiotensin II, bradykinin, substance P, acetylcholine, 5-HT or histamine. Pretreatment of the fundus strip with verapamil 0.3-1 microM antagonized in a concentration dependent fashion the neurotensin-induced contractions but not the muscular contractions caused by acetylcholine. It is concluded that neurotensin activates a specific excitatory receptor probably located on the cell membrane of the smooth muscles of the rat fundus. In addition, we suggest that this receptor is somehow related to a voltage-dependent calcium channel, sensitive to verapamil. PMID- 2988684 TI - On the type of receptor involved in the inhibitory action of adenosine at the neuromuscular junction. AB - The effects of adenosine and adenosine analogues (L-N6-phenylisopropyladenosine (L-PIA), D-N6-phenylisopropyladenosine (D-PIA), N6-cyclohexyladenosine (CHA), N6 methyladenosine, 5'-N-ethylcarboxamide adenosine (NECA) and 2-chloroadenosine) on evoked endplate potentials (e.p.ps) and on twitch tension were investigated in innervated sartorius muscles of the frog. Adenosine and its analogues decreased, in a concentration-dependent manner, the amplitude of both the e.p.ps and the twitch responses evoked by indirect stimulation. The order of potencies in decreasing twitch tension was: L-PIA, CHA, NECA greater than 2-chloroadenosine greater than D-PIA greater than N6-methyladenosine, adenosine. L-PIA was about ten fold more potent than D-PIA. None of the adenosine analogues tested affected the twitch responses of directly stimulated tubocurarine-paralyzed muscles. In concentrations that did not modify neuromuscular transmission, theophylline and 8 phenyltheophylline (8-PT) but not isobutylmethylxanthine (IBMX), antagonized the inhibitory action of 2-chloroadenosine at the neuromuscular junction. 8-PT behaved as a competitive antagonist and was about forty fold more potent than theophylline. It is concluded that the R-type adenosine receptor at the neuromuscular junction should not be classified in the A1/A2 system. The possibility of calcium-linked adenosine receptors having pharmacological profiles distinct from those originally defined as modulating adenylate cyclase is discussed. PMID- 2988686 TI - Biochemical studies in paraplegic renal stone patients. 2. Urinary excretion of citrate, inorganic pyrophosphate, silicate and urate. AB - The urinary excretions of citrate, inorganic orthophosphate and pyrophosphate, silicate and urate were measured in 17 paraplegic patients with renal calculous disease associated with urinary infection, in 16 paraplegics with no history of urinary calculus and in 14 healthy control subjects. The paraplegics excreted less citrate, orthophosphate and pyrophosphate than the control subjects. The stone-formers excreted more urate than the paraplegics without stone disease, but less than the control subjects. There were significant positive correlations between urinary orthophosphate and pyrophosphate and between urinary silicate and 24-h urine volume. The possible roles of increased urate and diminished excretion of citrate and inorganic pyrophosphate in the aetiology of renal calculosis in paraplegic patients deserve further investigation. PMID- 2988685 TI - The use of [3H]-[D-Pen2,D-Pen5]enkephalin as a highly selective ligand for the delta-binding site. AB - The characteristics of the binding of [3H]-[D-Pen2,D-Pen5]enkephalin were determined in homogenates of guinea-pig and rat brain. In the guinea-pig, the maximum binding capacity for [3H]-[D-Pen2,D-Pen5]enkephalin was 4.19 pmol g-1 and the KD 1.61 nM. In the rat, the corresponding values were 2.47 pmol g-1 and 5.42 nM. In both species, the maximum binding capacity and the affinity were not altered when mu-binding was suppressed with [D-Ala2,MePhe4,Gly-ol5]enkephalin. The mu-agonists, [D-Ala2,MePhe4,Gly-ol5]enkephalin and morphine, displaced a small portion of the binding of [3H]-[D-Pen2,D-Pen5]enkephalin with high affinities. PMID- 2988688 TI - Acute appendicitis and dietary fibre. PMID- 2988687 TI - Initial dose of enalapril in hypertension. PMID- 2988689 TI - HTLV-III antibodies in a remote population of eastern Zaire. PMID- 2988691 TI - T cell leukemia/lymphoma in Trinidad. PMID- 2988690 TI - Seroconversion of human T cell lymphotrophic virus III (HTLV-III) in patients with haemophilia: a longitudinal study. PMID- 2988692 TI - HTLV-III, haemophilia, and blood transfusion. PMID- 2988693 TI - Autoradiographic quantitation of beta-adrenergic receptors on neural cells in primary cultures. I. Pharmacological studies of [125I] pindolol binding of individual astroglial cells. AB - As part of an investigation into the expression of neurotransmitter receptors on astroglia, we have developed a method to label beta-adrenergic receptors on immunocytochemically stained cultured cells using autoradiography of 125I labelled beta-adrenergic receptor ligands. The current investigation was undertaken to determine whether the binding of [125I]pindolol (*IPIN) to immunocytochemically stained cultured cells, as measured by quantitative autoradiography, would fulfill the usual pharmacological criteria for specific beta-adrenergic receptor binding. *IPIN binding experiments were carried out on individual astroglia obtained from neonatal rat cerebral cortex and grown as primary cultures on polylysine-coated glass slides. Autoradiographic silver grains on cells which stained for the intracellular astroglial marker, glial fibrillary acidic protein (GFAP), were quantified by a microcomputer-based video digitizing system. We determined that competition for *IPIN binding by propranolol and isoproterenol was stereospecific, that specific binding of *IPIN was saturable, and that *IPIN binding sites were lost after isoproterenol-induced desensitization. These results provide convincing evidence that we can quantitatively examine beta-adrenergic receptors on single cells. The use of computerized video methods greatly facilitates the rapid quantitation of autoradiographic grains associated with immunocytochemically identified cells. This study is a unique demonstration of receptor binding parameters derived from single cells in a known population, and represents a novel approach to the problem of assessing cell-type specific receptors on neural cells in mixed primary cultures. PMID- 2988694 TI - Autoradiographic quantitation of beta-adrenergic receptors on neural cells in primary cultures. II. Comparison of receptors on various types of immunocytochemically identified cells. AB - We have developed a microcomputer-based video method to quantify neurotransmitter receptors on single, immunocytochemically labeled cultured cells. This method has been applied to determine whether beta-adrenergic receptors are more numerous on neurons, astroglia, oligodendroglia or fibroblasts in primary neural cell cultures, and to assess the heterogeneity of receptor expression within a single cell type. Dissociated cells from perinatal rat cerebral cortex were grown in very sparse cultures on polylysine-coated glass slides. The cultured cells were fixed and permeated, then stained with fluorescently labeled immunocytochemical markers for astroglia (glial fibrillary acidic protein), fibroblasts (fibronectin), oligodendroglia (galactocerebroside) or neurons (A2B5). beta Adrenergic receptors were labeled with [125I]pindolol or [125I]cyanopindolol, and dry-mount autoradiography was carried out on the fixed cells. Cells were identified according to their morphology and cell-type specific staining, then autoradiographic grains associated with the defined cells were visualized by reflected polarized light microscopy and counted with a microcomputer-based video digitizing system. Using this technique, we have determined that fibroblasts have less than 15% of the number of beta-adrenergic receptors expressed by polygonal astroglia, whereas oligodendroglia and neurons had no detectable binding of 125I labelled ligands. This suggests that in these mixed neural cell cultures, the great majority of beta-adrenergic receptors are associated with astroglia. Furthermore, we determined that process-bearing astroglia have less than 5% of the number of beta-adrenergic receptors expressed by polygonal astroglia. Since process-bearing astroglia are thought to be derived from polygonal astroglia, these results suggest that the beta-adrenergic receptor is lost from this population of astroglia during development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988695 TI - The formation of deaminated metabolites of dopamine in the locus coeruleus depends upon noradrenergic neuronal activity. AB - The effect of manipulations of noradrenergic neuronal activity on the levels of the deaminated metabolites of dopamine in the locus coeruleus has been investigated in the rat. Antidromic stimulation of the locus coeruleus increased the levels of 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 3-methoxy-4-hydroxyphenylethyleneglycol (MOPEG) in this area. Conversely, local infusion of tetrodotoxin into the locus coeruleus reduced the levels of these metabolites in this region. After systemic injection, idazoxan increased whereas clonidine diminished DOPAC, HVA and MOPEG levels in the locus coeruleus either in normal animals or in animals bearing a lesion of A9 and A10 dopaminergic cells. These results suggest that the formation of dopamine deaminated metabolites in noradrenergic cell bodies is dependent upon, and may serve as an index of, central noradrenergic neuronal activity. PMID- 2988697 TI - Hypothalamic epinephrine is released into hypophysial portal blood during stress. AB - The hypothalamic secretion of epinephrine (EPI), norepinephrine (NE), and dopamine (DA) into hypophysial portal blood was studied in adrenalectomized rats subjected to heat stress. Portal plasma EPI levels were increased 3-fold by heat stress whereas portal plasma concentrations of NE and DA were not altered by stress. These data suggest that EPI of hypothalamic origin may be involved directly in the modulation of anterior pituitary secretion during stress. PMID- 2988696 TI - Light-dependent regulation of dopamine receptors in mammalian retina. AB - The specific binding of [3H]spiperone, a D-2 dopamine receptor ligand, in in retinas from rabbits kept one week in constant light was significantly lower than in retinas from rabbits exposed to constant dark. Constant light did not alter the binding of [3H]spiperone in the striatum, where melatonin does not inhibit dopamine release. The decrease in [3H]spiperone binding induced by constant light in retina appears to be associated with the activation of inhibitory melatonin receptors on dopaminergic neurons. In support of this hypothesis, treatments that elevate melatonin concentrations, such as dark or melatonin administration, reversed the light-induced down-regulation of D-2 dopamine binding sites in retina. It is concluded that the decrease in melatonin levels in constant light disinhibits the dopamine-containing retinal neurons in vivo leading to elevated dopamine release and subsequent D-2 dopamine receptor down-regulation. PMID- 2988698 TI - Inputs from the A1 noradrenergic region to hypothalamic paraventricular neurons in the rat. AB - Electrical stimulation of the rat A1 noradrenergic region produced excitation (77%) of the activity of putative vasopressin (VP)-secreting neurons in the paraventricular nucleus (PVN) and produced excitation (4%), inhibition (26%) and excitation-inhibition (11%) of the activity of PVN neurons that were not antidromically identified by neurohypophysial stimulation. The excitatory response of putative VP-secreting neurons was blocked by microiontophoretically applied phentolamine, an alpha-adrenoceptor antagonist, but not by timolol, a beta-adrenoceptor antagonist. The inhibitory response of unidentified PVN neurons, on the other hand, was blocked by timolol, but not by phentolamine. PMID- 2988699 TI - Brain benzodiazepine receptor binding and purine concentration in Lesch-Nyhan syndrome. AB - Benzodiazepine receptor [( 3H]flunitrazepam) binding and purine concentration were measured in autopsied cerebral cortex of 4 patients who died with Lesch Nyhan syndrome. Receptor density was normal in all 4 regions of Lesch-Nyhan cortex examined. However, an enhancement of benzodiazepine receptor affinity (25% reduction in Kd) was found in well-washed parietal and occipital cortex homogenates. Maximal gamma-aminobutyric acid (GABA) stimulation of [3H]flunitrazepam binding was normal in temporal, parietal and occipital cortex but markedly reduced (by 50-80%) in frontal cortex. Increased sensitivity to hypoxanthine inhibition (30% reduction in Ki) was also observed in parietal cortex. The concentrations of the purines hypoxanthine, xanthine and inosine in Lesch-Nyhan parietal cortex were about twice the values measured in control material matched for postmortem time. We suggest that the above-normal concentrations of purines estimated to be present in Lesch-Nyhan brain may be sufficient to significantly affect the ability of the benzodiazepine receptor to modulate GABA-mediated brain mechanisms. PMID- 2988700 TI - Characterization of the spinal adrenergic receptors mediating the spinal effects produced by the microinjection of morphine into the periaqueductal gray. AB - After the microinjection of morphine (5 micrograms/0.5 microliter) into the periaqueductal gray resulted in an increase in the hot-plate and tail-flick response latency of the unanesthetized rat, the alpha-adrenergic antagonists yohimbine, rauwolscine and corynanthine were given intrathecally. This treatment resulted in a dose-dependent reversal of the inhibition of the thermally evoked tail-flick reflex. The relative potency of these stereoisomers was: yohimbine = rauwolscine greater than corynanthine. Given the reported affinity of these agonists for the alpha 2 (yohimbine/rauwolscine) and alpha 1 (corynanthine) receptors, these observations suggest that the spinopetal noradrenergic systems are acting on alpha 2-adrenergic receptors. Prazosin, an agent with several orders of magnitude higher affinity for the alpha 1 than the alpha 2-receptor, was at best only equiactive with yohimbine. None of the intrathecal treatments produced a significant reversal of the effects of periaqueductal gray morphine on the hot-plate response. This suggests that the activation of spinopetal noradrenergic pathways alone cannot account for the suppression by morphine in the periaqueductal gray of this response which is organized at the supraspinal level. PMID- 2988701 TI - Adrenocorticotropin immunoreactivity is contained within presumptive endocytotic vesicles of rat Brunner's gland cells. AB - Using an immunofluorescence microscopic staining technique, adrenocorticotropin (ACTH)-stained myenteric plexus perikarya and nerve fibers as well as ACTH immunoreactive submucous plexus nerve processes were revealed in the rat duodenum. However, ACTH-immunostained cells were also seen in Bunner's glands. In immunoelectron microscopic experiments could be demonstrated that the ACTH immunoreactivity was contained within presumptive endocytotic vesicles of these cells. The ACTH-positive vesicles had a mean diameter of 270 nm. The ACTH peroxidase anti-peroxidase (PAP) complex (mean diameter 50 nm) was located on the inner surface of the vesicle. At light microscopic level, ACTH-immunofluorescent nerve fibers were in close association with these ACTH-stained Brunner's gland cells. These findings might indicate that ACTH influences both the quality and quantity of the mucous produced by Brunner's gland cells. PMID- 2988702 TI - Tritium-sensitive film autoradiography of guinea pig brain alpha 2-noradrenergic receptors. AB - Tritium-sensitive film autoradiography was used to determine the distribution of alpha2-noradrenergic receptors (i.e. [3H]p-aminoclonidine binding sites) in guinea pig forebrain. Alpha 2-Receptors are heterogeneously distributed throughout the forebrain. Many limbic system structures, such as bed nucleus of stria terminalis, medial preoptic area, medial amygdaloid nucleus and lacunosum molecular layer in hippocampus were heavily labeled. We did not quantify receptor density in areas containing principally white matter but the optical density in those areas was similar to film background suggesting a very low receptor density. Low receptor concentrations were also found in areas that do not contain a high percentage of white matter, such as lateral septum and ventromedial hypothalamic nucleus. PMID- 2988703 TI - Estradiol modulation of alpha 2-noradrenergic receptors in guinea pig brain assessed by tritium-sensitive film autoradiography. AB - In this experiment, we examined the influence of estradiol on alpha 2 noradrenergic receptor binding in the female guinea pig brain with tritium sensitive film autoradiography. Treatment of ovariectomized guinea pigs with estradiol increased alpha 2-receptor binding in regions of the preoptic area and decreased binding in the ventromedial nucleus of the hypothalamus. There was no effect of estradiol on alpha 2-receptor concentrations in other estradiol concentrating regions or in brain areas which do not concentrate estradiol. PMID- 2988705 TI - Dependence of Na+,K+-ATPase and electrogenic component of Em in cultured myotubes on cell fusion. AB - This study was undertaken in order to determine the relation among cell fusion, [3H]ouabain binding and the membrane potential (Em) of cultured rat skeletal muscle. The amount of ouabain bound and the Em both increased with age, the increases being most dramatic following fusion. Inhibition of fusion prevented the developmental increases in both properties of cultured muscle. After fusion, the size of the electrogenic component of Em, determined by the decrease in Em produced by ouabain within 5-10 min, increased independent of the age at which fusion occurred. It is concluded that the increase in Em with age depends on postfusion appearance and activity of Na,K-ATPase. PMID- 2988704 TI - Blockade of the diazepam-induced increase in rat striatal acetylcholine content by the specific benzodiazepine antagonists ethyl-beta-carboline-3-carboxylate and Ro 15-1788. AB - Diazepam increased the acetylcholine content in the striatum and the hippocampus of the rat. This effect was antagonized in both brain areas by treatment with the specific central benzodiazepine blockers ethyl-beta-carboline-3-carboxylate and Ro 15-1788, whereas the peripheral antagonist Ro 5-4864 was ineffective. Pretreatment with picrotoxin, a known GABA antagonist did not interfere with the diazepam-induced acetylcholine increase. These results indicate a specific involvement of benzodiazepine receptors in the cholinergic action of diazepam and this effect appears to be independent of GABA receptor activation. PMID- 2988706 TI - Activation of NMDA receptors elicits fictive locomotion and bistable membrane properties in the lamprey spinal cord. AB - The motor pattern underlying locomotion in the lamprey can be elicited in the spinal cord in vitro by applying excitatory amino acids that activate NMDA receptors. When this is done oscillatory membrane potentials phase-linked with the locomotory rhythm can be recorded in different types of neurones. In some spinal neurones large amplitude oscillation continues after elimination of synaptic input with application of TTX. This oscillatory pacemaker-like activity is dependent on an activation of NMDA receptors, and is probably important in the generation of locomotion. PMID- 2988707 TI - Opioid inhibition of reflex urinary bladder contractions: dissociation of supraspinal and spinal mechanisms. AB - The supraspinal and spinal mechanisms of morphine induced inhibition of reflex urinary bladder contractions were studied in the urethane anesthetized rat. The inhibition of bladder contractions by intracerebroventricular (i.c.v.) morphine was abolished by intrathecal (i.t.) naloxone. In addition systemic reserpine (3.5 5.0 mg/kg, i.p.) abolished the inhibitory effect of both systemic morphine and i.c.v. morphine but not that of i.t. morphine. These data support the involvement of separate supraspinal and spinal mechanisms in the effects of morphine on bladder motility. The supraspinal effect appeared to be mediated indirectly via a monoaminergic system whereas the spinal action was mediated independently of monoamines. However, both the indirect supraspinal and the direct spinal mechanism involved a spinal naloxone-sensitive process possibly an endogenous enkephalinergic system. PMID- 2988708 TI - Specific [3H]gamma-aminobutyric acid binding to vestibular membranes of the chick inner ear. AB - To support a postulated neurotransmitter character of gamma-aminobutyric acid (GABA) in the vertebrate vestibule, [3H]GABA binding was measured in a crude membrane preparation of chick inner ear ampullary cristae. In the absence of divalent cations bound [3H]GABA was displaced by unlabeled GABA, muscimol or bicuculline, but it was not displaced by (+/-)-baclofen. A single population of [3H]GABA binding sites with an equilibrium constant of 19.4 nM and a maximum binding capacity of 0.58 pmol/mg protein was found. These results suggest the possible existence of a synaptic GABAA receptor in the chick inner ear membranes and sustain the neurotransmitter role of GABA in the chick vestibule. PMID- 2988709 TI - The ontogenetic development of neurotensin-like immunoreactivity and neurotensin receptors in the cat striatum. AB - At birth, striatal neurotensin-like immunoreactivity amounted to 10% of the adult values which were reached at the age of 5 weeks. In the caudate nucleus neurotensin-like immunoreactivity presented a patchy distribution throughout development that was in register with Met-enkephalin staining, whereas [3H]neurotensin binding sites were most heavily concentrated in the background matrix. Thus, the adult distribution pattern of neurotensin-like immunoreactivity and [3H]neurotensin binding sites is already established at birth. PMID- 2988710 TI - Immunocytochemical localization of 5'-nucleotidase in oligodendroglia and myelinated fibers in the central nervous system of adult and young rats. AB - Rat central nervous system (CNS) tissue sections were immunostained by the peroxidase-anti-peroxidase (PAP) method using a rabbit serum directed against rat liver 5'-nucleotidase. In paraffin sections from the brains of 60-day-old rats 5' nucleotidase immunoreactivity occurred in the same white-matter regions as myelin basic protein immunoreactivity and histological staining of myelin. The immunostaining of cerebral white matter for 5'-nucleotidase was more intense and wide-spread at the age of 120 days than at 60 days, and the choroid plexus and blood vessels were stained consistently. In the paraffin sections from the brains of younger (20-day-old) rats the staining of 5'-nucleotidase in the white matter was faint and patchy. In paraffin sections from spinal cord, 5'-nucleotidase immunoreactivity was observed throughout the lateral white-matter columns and, frequently, in the cell bodies of interfascicular oligodendroglia. Interfascicular oligodendroglia also showed 5'-nucleotidase immunoreactivity in vibratome sections from the CNS tissue of young and adult rats. The findings were consistent with histochemical and biochemical evidence for 5'-nucleotidase in rat brain myelin and oligodendroglia, with substantial increases in activity in the myelin as rats develop from the ages of 20 to 120 days. 5'-Nucleotidase immunoreactivity was not observed in any astrocytes or in oligodendrocytes in the gray matter; however, the enzyme may occur in those glial cells at levels lower than were detectable using the present method. PMID- 2988711 TI - Blockade of adrenal compensatory hypertrophy by unilateral hypothalamic lesions. AB - To determine if unilateral hypothalamic lesions block adrenal compensatory hypertrophy (ACH), such lesions were produced unilaterally under stereotaxic guidance using direct cathodal current in rats just after the left adrenal gland was removed. The animals were sacrificed one week later and the degree of the ACH of the remaining adrenal gland was determined. Animals without lesions uniformly showed ACH in terms of an absolute increase in adrenal weight of the remaining gland as compared to the one excised or if calculated as mg adrenal weight/100 g body weight. On the other hand, lesions in the region lateral to the paraventricular nucleus, lateral to the median eminence or in the region between these two loci prevented ACH in roughly 50% of the animals whether the lesion was placed on the same or opposite side of the hypothalamus. Some lesions in the mid line in the vicinity of the paraventricular nucleus also resulted in blockade of ACH. We speculate that the blockade is due to a partial elimination of CRF and/or vasopressin/oxytocinergic pathways which are activated by the fall in plasma corticosterone after removal of an adrenal gland. PMID- 2988712 TI - Effects of guanyl nucleotides and ions on kappa opioid binding. AB - Displacement curve analyses demonstrated that GTP and its nonhydrolyzable analog, GPP(NH)P, inhibited the binding of [3H]dihydromorphine (mu agonist), [3H]D-Ala2-D Leu5-enkephalin (delta agonist), and [3H]ethylketocyclazocine (general agonist) but not [3H]diprenorphine (general antagonist). Using a paradigm to block mu and delta sites with specific cold ligands, [3H]ethylketocyclazocine labeled kappa sites which were less GTP sensitive than sites labeled by mu and delta agonists. Further, Na+ and Mg++, important in inhibitory adenylate cyclase systems, also decreased both unblocked and mu-/delta-blocked [3H]ethylketocyclazocine binding. Scatchard analyses revealed that the inhibitory effects of GTP result in a decrease in affinity without a significant change in binding capacity, and that the kappa component of [3H]ethylketocyclazocine binding was less sensitive to the effects of GTP than binding sites labeled by mu or delta agonists. In comparison to the effects of GTP, Na+ decreased binding affinity but increased the binding capacity of the kappa component. These data also suggest that the inhibitory effects of Na+ and GTP on binding affinity are not additive. Association and dissociation plots revealed that although both components of binding may be involved in these affinity changes, the dissociation rate represents the more significant factor. These data suggest that [3H]ethylketocyclazocine binding to kappa sites is GTP and Na+ sensitive. However, it should be noted that [3H]ethylketocyclazocine binding to kappa sites is less sensitive to GTP than its binding to other opiate sites, and that this kappa binding is differentially affected by Na+. The significance of these characteristics with regard to the effect of kappa opiates on adenylate cyclase activity remains to be determined. PMID- 2988714 TI - [Establishment of five human pulmonary carcinoma cell lines and their biologic properties]. PMID- 2988713 TI - [Characterization of the effects of a derivative of pyridoindole (DH-1011) on alpha2-adrenergic membrane receptors in the smooth muscle of mesenteric arteries and coronary arteries]. PMID- 2988715 TI - [Advances in the study of JC and BK viral infections]. PMID- 2988716 TI - Calcium-acidic phospholipid-phosphate complexes in human atherosclerotic aortas. AB - Since aortic calcification chemically resembles bone mineralization, we tested the hypothesis that a bone mineral nucleator, the Ca-acidic phospholipid phosphate complexes, is found in atherosclerotic plaques. Calcium-acidic phospholipid-phosphate complexes were isolated from hydroxyapatite-containing calcified plaques and from adjacent nonmineralized areas of adult human aortas. Neonatal aortas, which served as nonmineralized lesion-free controls, contained negligible amounts of the Ca-acidic phospholipid-phosphate complexes. The concentration of complexed acidic phospholipids in the aortic plaques (5 micrograms/mg demineralized dry wt) was comparable to that found in newly mineralized bone and calcified cartilage. The presence of Ca-acidic phospholipid phosphate complexes in the nonmineralized regions of the adult aorta, as well as in the mineralized plaques, suggests that this tissue may calcify through mechanisms similar to those involved in bone mineralization. PMID- 2988717 TI - The precipitation of calcium carbonate polymorphs in vitro at 37 degrees C. AB - In order to determine some of the factors that affect the formation of the three polymorphs of calcium carbonate in gallstones, its precipitation from undisturbed solutions containing calcium chloride and sodium bicarbonate was studied. The variation in the amount of calcium carbonate formed with time was studied by weighing the precipitate, and the percentages of calcite, vaterite, and aragonite were determined by x-ray diffraction (XRD) analysis. The effects of additives and stirring were noted. Minor changes in conditions were sufficient to affect the formation of the polymorphs, and all three polymorphs could be found simultaneously in some solutions. It is suggested that no great differences in precipitating conditions need to be postulated to account for the presence of different polymorphs in gallstones. Aragonite seems to be formed slowly in undisturbed conditions. PMID- 2988718 TI - Pulsed electromagnetic fields promote collagen production in bone marrow fibroblasts via athermal mechanisms. AB - Primary and passaged cultures of fibroblasts (RBMFs) raised from the bone marrow stroma of young rabbits were treated with pulsed electromagnetic fields (PEMFs) from the start of each culture until 1 week after they became confluent. the PEMF treatment had no effect on cell proliferation, estimated by phase contrast microscopy, by 3H-thymidine incorporation into DNA, or by total DNA assay. Collagen production, estimated by conversion of 3H-proline to 3H-hydroxyproline in nondialyzable material was markedly elevated in postconfluent cultures, but not in cultures that had only just reached confluence. About 65 of 3H hydroxyproline was in low molecular weight form, and a correlation between collagen breakdown and cyclic AMP (cAMP) levels in RBMFs was demonstrated by adding dibutyryl cAMP or prostaglandin E3 (PGE2) to the culture medium concurrently with 3H-proline. The PEMF apparatus caused an insufficient temperature rise (less than 0.1 degree C) to account for these results. We propose that the rise in collagen production is consistent with the hypothesis that PEMFs act by reducing cAMP levels in RBMFs, and that thermal effects are insignificant. PMID- 2988719 TI - Properties of Suc-GPLGP-MCAase and dipeptidyl-aminopeptidase in mouse calvaria derived osteoblastic cells (MC3T3-E1). AB - In this study the properties of Suc-GPLGP-MCAase and dipeptidyl-aminopeptidase (DAP) in clone MC3T3-E1 cells which have osteoblastic ability were examined. The Suc-GPLGP-MCAase was the most active at pH 8.0 and its molecular weight was about 65,000 as judged by gel filtration method. The enzyme activity was increased by some metal ions such as Mn+2, Ca+2, and Mg+2 but inhibited by Zn+2 and Cu+2. EDTA increased the enzyme activity to 23-fold. The enzyme activity was slightly inhibited by thiol inhibitors, N-ethylmaleimide, and p-chloromercuribenzoic acid by 16% and 27%, respectively. The DAP has an optimum pH at 7.5, and a molecular weight of about 100,000. The enzyme was completely inhibited by diisopropylfuorophosphate, but not by N-ethylmaleimide, iodoacetic acid, p chloromercuribenzoic acid, phenylmethylsulfonyl fluoride, EDTA, and several metals. These results show that Suc-GPLGP-MCAase and DAP in MC3T3-E1 cells have different novel properties compared with these enzymes in the peripheral tissues. PMID- 2988720 TI - Ethanol-induced formation of solid calcium phosphates. AB - Conventional transmission electron microscopy requires an effective dehydration of the tissue specimens under investigation. This dehydration, for which ethanol most often is used, may influence the solubility of the slightly soluble calcium phosphates. To study the behavior of calcium phosphates in ethanol, stable neutral aqueous solutions containing the ions were mixed with ethanol in various ratios at 20 degrees C. After 24 hours, the precipitates formed wee identified by x-ray diffraction to be either apatite, brushite, or amorphous calcium phosphate. The results are discussed with reference to possible formation of calcium phosphate artifacts. PMID- 2988722 TI - The herpes affliction. PMID- 2988721 TI - Hydroxylapatite and skin graft vestibuloplasty. A new dimension in preprosthetic surgery. PMID- 2988723 TI - Evidence for oxytocin receptors in the urinary bladder of the rabbit. AB - Experiments were performed on isolated detrusor smooth muscle from New Zealand White rabbits. Oxytocin was shown to exhibit high intrinsic contractile activity on isolated strips of detrusor muscle, where the maximum contractile amplitude was 12% greater than control responses to 1 microM carbachol. Repeated applications of 1 microM oxytocin were associated with tachyphylaxis representing a 49% decrease in the amplitude which became reproducible after several applications without further decay of contractile strength. Dose-response experiments indicated that threshold contractions to oxytocin occur at 3 nM and were maximum at 10 microM with mean effective concentration of 125 nM. The contractile responses to 1 microM oxytocin were not antagonized by phentolamine, atropine, methysergide, saralasin, or naloxone, but were partially inhibited by 1 microM of indomethacin. Ligand binding studies on partially purified membrane preparations from detrusor smooth muscle were performed over a range of 78 pM to 10 nM with 125I-labelled oxytocin. Scatchard analysis of specific bound receptors indicated a KD of 2.5 nM and Bmax of 187 fmol/mg protein and a second compartment that was unsaturable at the concentrations of ligand employed. Nonspecific binding ranged from 36 to 77% of the total binding. PMID- 2988724 TI - Ionic permeability and blood pressure. AB - The difficulty of establishing a causal relationship between any measured phenomenon and hypertension is emphasized. The literature concerning ion movements in tissues from hypertensive patients and animals is briefly reviewed. It is pointed out that the evidence (i) is conflicting concerning changes of permeability of red blood cells in patients with hypertension; (ii) is more consistent in leukocytes; and (iii) is virtually nonexistent regarding ionic permeability of blood vessels from man. A few studies suggest that permeability to calcium is altered in aortae from hypertensive animals and that concentrations of calcium in platelets are correlated with blood pressure in man. Some of our previously published work on red blood cells is then summarized which suggests that altered passive permeability to calcium exists in red blood cells from spontaneously hypertensive rats. Experiments are described using the calcium antagonists D-600 or felodipine to further study previously reported differences in calcium handling in blood vessels from spontaneously hypertensive rats. We could detect no difference in the effects of these calcium antagonists on blood vessels from hypertensive animals compared with controls. It is suggested that the finding of increased passive permeability to calcium in tissues from hypertensive animals is a lead worth pursuing. PMID- 2988725 TI - Primary versus secondary events in hypertension. AB - Functional modifications, such as a reduction in hormonal response, which occur in the cardiovascular system in hypertension, are reflected at the cellular level by anomalies in cyclic nucleotide and other messenger systems. To distinguish between primary and adaptive abnormalities, we pursued three research strategies. (i) Investigations on various models of hypertension. To be considered a primary defect, an abnormality should also be present in other genetically hypertensive models. Indeed, we have confirmed the occurrence of cellular hyperplasia in the heart of spontaneously hypertensive rats (SHR) as well as in spontaneously hypertensive mice (SHM). An increase of calmodulin levels in the heart and kidney is also observed in both the SHR and SHM. (ii) Studies on the evolution of hypertension with age. In humans, a decrease of cAMP levels in response to beta adrenergic stimulation in older patients is contrasted with an excess in younger subjects. In the SHR, protein kinase activity of the heart is lower in the prehypertensive stages, whereas this defect appears much later in the aorta. (iii) Experiments on anomalies in newborns and cultured cells. The heart and kidney in the SHR exhibit significant hyperplasia at birth, and an abnormal growth continues in tissue culture. We hope that these strategies will eventually help to provide biochemical and functional markers for genetic analysis of factors which may be involved in the pathogenesis of hypertension. PMID- 2988727 TI - Transformation of hamster embryo cells by chymotrypsin-treated and untreated polyoma virus: characterization of transformants. AB - The ability of chymotrypsin-treated (chymo+) and untreated (chymo-) polyoma virus to transform cultured hamster embryo fibroblasts was examined. The data show that exposure to this protease reduces the ability of the virus to transform non permissive cells to essentially the same extent as it reduces its ability to replicate in permissive cells. Twenty-five lines of transformed cells were established from colonies growing in soft agar, and after 20 in vitro passages, cells of all lines were characterized with respect to their ability to form colonies in soft agar and their tumorigenicity in hamsters. While the studies showed that there are striking differences among the lines with respect to colony forming ability, and real, though less striking differences in tumorigenicity, they failed to reveal any obvious differences between the groups of cell lines transformed by chymo- and chymo+ polyoma virus. Of 13 lines examined, all were found to express both middle and small polyoma T antigens, none express significant levels of large T antigen, and 11 express some form of what is probably a truncated large T antigen, the most common species having a molecular weight of 67000. PMID- 2988726 TI - Genetic linkage but independent expression of functional HSV-1 tk and mammalian aprt genes after cotransfer to L cells. AB - DNA-mediated gene transformation of mouse Ltk-aprt-hprt-cells was used to obtain stable, doubly selected transformants simultaneously expressing herpes virus thymidine kinase (TK) and mammalian adenine phosphoribosyltransferase (APRT). Cotransformants occurred at a frequency of 5 X 10(-6), a similar frequency for the transfer of the aprt marker has been previously observed. Isozyme and Southern blot analysis show that the TK and APRT expressed in these transformants resulted from gene transfer. For one stable cotransformant, [3H]thymidine [( 3H]TdR) selection against TK activity resulted in the loss of APRT activity as well, suggesting that these genes had become genetically linked together. Similarly selection against APRT expression resulted in the loss of a subset of the transferred herpes simplex virus tk genes. 5-Bromodeoxyuridine (BUdR) selected TK- variants differed from [3H]TdR selected TK- variants, in that they retained tk genes. However, BUdR-selected variants expressed full levels of APRT. Therefore, even though the transferred tk and aprt genes had become genetically linked together, they were, in this case, independently expressed since these cells were phenotypically TK- and APRT+. PMID- 2988728 TI - The contribution of exoproducts to virulence of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa produces a large number of extracellular products which may contribute to its virulence. We have employed a genetic approach to determine the contribution of toxin A, exoenzyme S, elastase and alkaline protease to the pathogenesis of P. aeruginosa. Mutations have been introduced with chemicals or transposons. Mutants have been identified using immunological, chemical, or toxicity assays. Mutants were extensively characterized in vitro to ascertain that they were identical to their parent strain except for the production of the desired product. Appropriate mutants were compared with their parent strains in several animal models: the burned mouse model, the mouse corneal infection model, and a rat model of chronic lung infection. The data indicate that virulence of P. aeruginosa is multifactorial. Further, the relative contribution of a given P. aeruginosa product may vary with the type of infection. PMID- 2988729 TI - The Vancouver Lymphadenopathy-AIDS Study: 2. Seroepidemiology of HTLV-III antibody. AB - Testing for antibody to human T-lymphotropic retrovirus (HTLV-III) was carried out in 448 participants in the Vancouver Lymphadenopathy-AIDS (acquired immune deficiency syndrome) Study. The overall prevalence rate of seropositivity was 34%. Of 130 seronegative subjects followed for an average of 8.5 months, 14 became seropositive; thus, the approximate annual seroconversion rate was 15%. More than 100 male sexual partners in one's lifetime, frequent receptive anal intercourse, fisting, a history of gonorrhea or hepatitis, and frequent sexual contact in clubs were found to be independent risk factors for HTLV-III seropositivity. PMID- 2988731 TI - Clonidine in spinal cord injury. PMID- 2988730 TI - The Vancouver Lymphadenopathy-AIDS Study: 3. Relation of HTLV-III seropositivity, immune status and lymphadenopathy. AB - In a study of 394 homosexual men recruited at the primary care level the prevalence of antibody to human T-lymphotropic retrovirus (HTLV-III) was higher among those with lymph node enlargement than among controls. The degree of abnormal immune function, as shown by abnormalities in immunoglobulin levels, immune complex activity and T-lymphocyte subsets, was correlated with the extent of lymphadenopathy. A similar pattern of immunologic abnormality was associated with seropositivity for HTLV-III antibody. However, HTLV-III seropositivity was the major determinant of immune function after adjustment for lymph node status. The results suggest that the immune dysfunction seen in patients with lymphadenopathy is due for the most part to the high prevalence of HTLV-III seropositivity in these populations. Lymphadenopathy, in many subjects, may in fact represent a physical sign of a stabilized compensated homeostatic host response. Factors responsible for severe immune decompensation associated with acquired immune deficiency syndrome (AIDS) may best be sought by prospective study of HTLV-III seropositive asymptomatic patients or those with stable persistent generalized lymphadenopathy and relatively normal immune function. PMID- 2988732 TI - Relation of HTLV-III seropositivity and lymphadenopathy. AB - Testing for antibody to human T-lymphotropic retrovirus (HTLV-III) was carried out in five groups of homosexual men: 250 without lymphadenopathy (control group), 37 with slight or nonpersistent lymph node enlargement (intermediate group), 141 with persistent generalized lymphadenopathy, 32 with persistent generalized lymphadenopathy who underwent biopsy and 11 in whom acquired immune deficiency syndrome (AIDS) was diagnosed. The rates of HTLV-III seropositivity in the five groups were 18%, 32%, 61%, 94% and 91% respectively. PMID- 2988733 TI - Angiotensin converting enzyme. A possible histologic indicator for human renal cell carcinoma. AB - Angiotensin converting enzyme (ACE) was studied in specimens of human renal cell carcinoma by indirect immunofluorescence using rabbit antibody to human kidney ACE and fluorescein-labeled goat antibody to rabbit immunoglobulin. ACE was demonstrated in the cells of all renal cell carcinomas from 10 patients and in the cultured cells set up from 4 of them, but not in any cells of 12 extrarenal carcinomas of prostate, lung, and alimentary tract. These results suggest that immunohistologic detection of ACE is of value in the histologic diagnosis of renal cell carcinoma. PMID- 2988734 TI - Aspiration biopsy of the kidney. Simultaneous collection of cytologic and histologic specimens. AB - Aspiration biopsy of mass lesions of the kidney is a diagnostic procedure whose potential benefit to patients with equivocal radiologic findings, suspected metastases, palpable flank masses, nonfunctioning kidneys and even cortical cysts has not been fully explored. Over the past 7 years 163 renal aspiration specimens from 152 patients have been examined. Diagnostic yield was enhanced by use of modified "fine" needles with notched tips so that tissue fragments for histology as well as smears for cytology were obtained in 89% of solid tumors. The availability of aspirated tissue contributed significantly to pathologic classification and often spared patients additional surgery for confirmation of the cytologic diagnosis. Among the 152 cases, there were 35 with renal cell carcinomas, a sufficient number for detailed semiquantitative evaluation of their cytologic features. The remaining cases comprised metastatic carcinomas, lymphomas, transitional cell carcinomas, oncocytomas, cortical cysts, and miscellaneous conditions, e.g., abscesses, nonfunctioning kidneys, and hematomas. Overall, the aspiration biopsy determined the nature of the renal mass in 141 cases (93%). False-negative interpretations were due to insufficient diagnostic material in all but one instance. There was one false-positive result. PMID- 2988736 TI - Treatment of malignant fibrous histiocytoma of bone. A plea for primary chemotherapy. AB - Seven patients have been treated for malignant fibrous histiocytoma (MFH) of bone since the end of 1977. One patient received no chemotherapy, and one did not complete attempted chemotherapy. Both died, 7 and 51 months after diagnosis, respectively. The remaining five patients completed chemotherapy. Two first underwent a primary amputation, whereas the other three received primary chemotherapy with histologic evaluation of the effect. These patients showed a complete remission. The five patients who completed chemotherapy are all still alive, without indications of metastases or local recurrence. Although the number of cases is small, a 25- to 58-months (mean, 45) survival, in five patients treated either with chemotherapy alone or chemotherapy and surgery, is surprisingly good in view of previous experience with this tumor. In some of these patients, the authors were able to document an absence of any viable tumor following chemotherapy. PMID- 2988735 TI - The specific diagnosis of hepatocellular carcinoma by scintigraphy. Multiple radiotracer approach. AB - The accuracy of scintigraphy in diagnosing hepatocellular carcinoma (HCC) at Boston City Hospital between January 1, 1978 and September 30, 1983 is retrospectively reviewed. A combined protocol using technetium-99m sulfur colloid (TsSC), gallium (Ga), and scintiangiography (STA) was employed in order to enhance diagnostic specificity. There were 14 cases of HCC, of which 10 were proven histologically. The others were diagnosed clinically and angiographically. With one exception, all patients who had triple tracer scintigraphy showed a specific pattern of findings: (1) cold defects with TcSC; (2) Ga-avid foci, and (3) increased vascular supply from hepatic arteries. One false-positive study and one false-negative study were originally reported, although in both cases, strict adherence to the three criteria above would have avoided diagnostic error. These results indicate that triple tracer scintigraphy may be an effective diagnostic test for HCC. The relative efficacy of scintigraphy, ultrasonography, and computerized tomography in diagnosing HCC is also discussed. PMID- 2988737 TI - Combined modality treatment of operated astrocytomas grade 3 and 4. A prospective and randomized study of misonidazole and radiotherapy with two different radiation schedules and subsequent CCNU chemotherapy. Stage II of a prospective multicenter trial of the Scandinavian Glioblastoma Study Group. AB - A prospective and randomized trial has been performed in order to evaluate combined modality therapy in patients with astrocytomas grade 3 and 4. Follow-up information is available on 244 patients. One half of the series received radiation therapy twice a week (40.00 Gy/5 weeks), the other half five times a week (50.00 Gy/5 weeks). Misonidazole 1.2 g/m2 was given orally to one half of the patients in the first radiation treatment group 3 1/2 to 4 hours before the treatment. The other half received placebo. The second radiation treatment group was also divided in two halves, one receiving 0.48 g/m2 misonidazole and the other placebo 3 1/2 to 4 hours before radiation. The randomization also included a subdivision of the material into eight groups of which four were given CCNU and four no chemotherapy, beginning 3 months after operation. The dose of CCNU was 120 mg/m2 body surface every 6 weeks. All eight treatment groups showed practically identical periods of median survival, and no statistically significant differences were observed with regard to performance status, side effects, or complications. Another dosage and timing of misonidazole administration in relation to the irradiation schedule, and a consideration of effects of concomitant drugs like dexamethasone and phenytoin are discussed. PMID- 2988738 TI - Combination chemotherapy with adriamycin, etoposide, and cyclophosphamide for small cell carcinoma of the lung. A study by the EORTC Lung Cancer Working Party (Belgium). AB - The current study reports the results of Adriamycin (doxorubicin), etoposide, and cyclophosphamide (AVE) in small cell bronchogenic carcinoma. The overall rate of response was 82% in patients with limited disease and 66% in patients with extensive disease; complete remissions have been achieved in 20% of the patients with limited disease and in 7% of those with extensive disease. The median duration of survival was 14 months in patients with limited disease and 8.3 months in those with extensive disease. The results, and the analysis of literature, suggest that survival rather than response should be used to compare studies of chemotherapy in small cell bronchogenic carcinoma. PMID- 2988739 TI - Doxorubicin hydrochloride (Adriamycin) cardiotoxicity evaluated by sequential radionuclide angiocardiography. AB - A prospective study was carried out to evaluate the role of radionuclide angiocardiography (CEF) in accessing subclinical cardiotoxicity secondary to Adriamycin (doxorubicin) therapy in 73 women with gynecologic malignancies. Based on the findings of this study, the authors conclude that all patients should have an initial CEF before Adriamycin therapy. In patients with an initial CEF of greater than or equal to 55, frequent determinations are not necessary unless there is a significant decrease from the initial CEF. Patients with low normal initial CEF or significant difference between the initial CEF and minimum CEF should have CEF studies performed at more frequent intervals. Patients who develop below normal CEF should have Adriamycin withheld and CEF should be repeated at more frequent intervals. If the CEF returns to normal Adriamycin therapy can be reinstated. Patients requiring continuation of Adriamycin past 550 mg/m2, can safely do so as long as the CEF values remain normal. PMID- 2988740 TI - Vincristine, dactinomycin, and cyclophosphamide in the treatment of malignant germ cell tumors of the ovary. A Gynecologic Oncology Group Study (a final report). AB - Seventy-six patients with malignant germ cell tumors of the ovary received vincristine, dactinomycin, and cyclophosphamide (VAC) postoperatively. Fifty-four were treated after removal of all gross disease. The majority of these remain disease-free. Indeed, only 15 (28%) have failed, including 11 of 24 with pure endodermal sinus tumor, 3 of 11 (27%) with mixed germ cell tumor containing endodermal sinus elements, and only 1 of 20 with immature teratoma grade 2 or 3, a patient seen initially with recurrent disease. Postoperative VAC therapy, however, did not appear to be effective in patients with unresectable or incompletely resected germ cell tumors of the ovary. Fifteen of 22 patients (68%) with incompletely resected germ cell tumors failed VAC therapy, including 4 of 7 with pure endodermal sinus tumor, 5 of 5 with mixed germ cell tumors containing endodermal sinus elements, 2 of 2 with embryonal carcinoma, and 4 of 8 with immature teratoma. In failing, patients' median time to progression was 8 months. Dose-limiting toxicity was seen in 30% of the entire group. Combined cisplatin, vinblastine and bleomycin therapy now is being tested in this group of tumors. PMID- 2988741 TI - Malignant pleural mesothelioma following radiotherapy in a 16-year-old boy. AB - A case of diffuse, epithelioid mesothelioma of the right pleura in a 16-year-old boy is presented. Though a history of previous exposure to asbestos could not be elicited, the patient did receive pulmonary irradiation for metastatic Wilms' tumor at the age of 2 years. The authors believe that the development of his mesothelioma was causally related to his prior radiotherapy. PMID- 2988742 TI - Localization of hepatitis B surface and core antigens in human hepatocellular carcinoma by immunoperoxidase methods. Replication of complete virions of carcinoma cells. AB - The localization of hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) was investigated by an indirect immunoperoxidase method in formalin-fixed, paraffin-embedded liver specimens obtained from 95 Japanese patients with hepatocellular carcinomas. Routine and immune electron microscopic examinations were done in one case. The correlation between expression of hepatitis B virus antigens in the tissue and serum hepatitis B e antigen (HBeAg)/antibody to HBeAg (anti-HBe) status was examined. Hepatitis B surface antigen was detected in the cytoplasm of noncarcinomatous hepatocytes in 28 (29.5%) cases and of carcinoma cells in 11 (11.6%) cases. Hepatitis B core antigen was stained in noncarcinomatous hepatocytes in 13 (13.7%) cases and in carcinoma cells in 4 (4.2%) cases. Hepatitis B core antigen was present mainly in the nuclei, and all HBcAg-positive cases were positive for HBsAg. The routine electron microscopic examination revealed many round particles, 25 to 30 nm in diameter both in the nuclei and in the cytoplasm, and larger particles, 40 to 45 nm in diameter in the cytoplasm of carcinoma cells. Both types of particles had reaction products of HBcAg by immunoelectron microscopic study. Therefore, it was confirmed that the former were cores and the latter were Dane particles. There was a tendency that HBeAg-seropositive cases showed localization of HBcAg in the noncarcinomatous tissue. Among four cases with positive HBcAg in carcinoma cells, two were positive for HBeAg, one was positive for anti-HBe, and the other was negative both for HBeAg and anti-HBe in the sera. The data suggested occasional production of complete hepatitis B viruses of carcinoma cells in anti-HBe-positive as well as in HBeAg-positive hepatocellular carcinomas. PMID- 2988743 TI - Malignant fibrous histiocytoma of soft tissue. An analysis of 78 cases located and deeply seated in the extremities. AB - Seventy-eight cases of malignant fibrous histiocytoma (MFH) of soft tissues are reported. All tumors were deeply located in the extremities. An analysis is presented with regard to sex, age, site, macro- and microscopic features. The staging was assessed according to Enneking's classification (IB, one case; IIA, 25 cases; IIB, 42 cases; III, 10 cases). The recurrence rate after surgical treatment was high (37.5%) with significant differences after inadequate surgery (70%), wide excision (38%), or amputation (0%). Adjunctive radiotherapy rarely was effective in preventing local recurrence. The overall 5-year survival rate was 36%. The prognosis was worse in (1) tumors larger than 5 cm in diameter, (2) tumors that recurred early (less than 1 year) after surgery, and (3) tumors that had inadequate surgical treatment (intralesional or marginal excision). PMID- 2988744 TI - Bone marrow morphologic changes after combination chemotherapy including VP-16. AB - Bone marrow aspirates and biopsy specimens were examined from seven patients with small cell lung cancer. All patients had received recent combination chemotherapy including VP-16. No correlation between marrow biopsy cellularity and hematologic toxicity could be established. However, an unusual combination of morphologic changes was seen. This included an initial rapid increase in the M:E ratio, interference with cell division, and eventual cell death. There was little evidence of mitotic arrest or megaloblastosis. These changes are consistent with the known mechanism of action of VP-16, the only agent given to all patients. The small sample size and the unknown contributions of the other cytotoxic agents administered allow only limited conclusions. PMID- 2988745 TI - Molecular and immunologic analysis of a chronic lymphocytic leukemia case with antibodies against human T-cell leukemia virus. AB - The human T-cell leukemia virus type-I (HTLV-I) is a unique, exogenous, horizontally transmitted retrovirus which is T-cell tropic, and has been associated with a specific type of aggressive leukemia/lymphoma of mature T-cell origin. In a survey of lymphoid malignancies in Jamaica, antibodies to HTLV-I were also found in 6 of 17 patients with chronic lymphocytic leukemia (CLL), raising the possibility of an etiologic relationship. Further studies were undertaken on one of these patients to clarify the nature of the disease and possible virus relationship. Cell surface marker analysis of her peripheral blood cells documented that the majority of circulating lymphocytes were B-cells. DNA cloned probe analysis with a complete HTLV-I proviral genome of these peripheral malignant B-cells, was negative for integrated virus. A T-cell line was established in culture from her peripheral blood. The presence of HTLV-I in the cultured T-cell line was established by the detection of expressed viral specific gag protein p-19 and proviral DNA. Thus, a B-cell lymphoid malignancy can occur in the presence of HTLV-I infected T-cells, suggesting the possibility of an indirect leukemogenic mechanism. PMID- 2988746 TI - Glandular or mucus-secreting components in squamous cell carcinoma of the esophagus. AB - A review of 195 patients with carcinoma of the esophagus disclosed 41 cases (21.0%) with glandular and/or mucus-secreting components, in addition to the ordinary component of squamous cell carcinoma. These tumors could be grouped into three types according to representative histologic features of glandular and mucus-secreting portions: glandular type (23 cases), cribriform type (11 cases), and mucoepidermoid type (7 cases). The histologic features of the three types were reminiscent of those of adenocarcinoma, adenoid cystic carcinoma, and mucoepidermoid carcinoma of salivary glands, respectively. Moreover, areas showing glandular or mucus-secreting differentiation were in greater part located in the submucosa and the lamina propria mucosae, thereby suggesting that such differentiation had arisen in the esophageal glands or their ducts. In all 41 cases, the ordinary element of squamous cell carcinoma, invasive, or noninvasive, was admixed in various proportions with the glandular components, indicating that this type of esophageal tumor had originated not only from the covering squamous epithelium but also from esophageal mucous-gland or ductal epithelium. The findings also support the concept of the field origin of carcinogenesis in esophageal carcinoma. PMID- 2988747 TI - The accelerated phase of Chediak-Higashi syndrome. An expression of the virus associated hemophagocytic syndrome? AB - The clinical and pathologic findings of four patients with Chediak-Higashi syndrome in the accelerated phase were studied in order to clarify the nature of this enigmatic process. Fever, lymphadenopathy, hepatosplenomegaly, and cytopenias were present in every patient. All cases demonstrated extensive parenchymal infiltrates in many organs composed of benign-appearing histiocytes manifesting hemophagocytosis accompanied by lymphocytes and plasma cells. Studies in one patient suggested a viral etiology with the findings of a low blood lymphocyte OKT4 to OKT8 ratio, acquired loss of lymphocyte response to mitogens, the presence of Epstein-Barr virus genome in the mononuclear cells of lymph node, blood, and bone marrow, and possible clinical responses to acyclovir. It is concluded that the accelerated phase of Chediak-Higashi syndrome may be the clinicopathologic expression of the virus-associated hemophagocytic syndrome. PMID- 2988748 TI - Serum creatine kinase BB isoenzyme as a diagnostic aid in occult small cell lung cancer. AB - Four patients with metastatic small cell carcinoma of the lung who had initial chest x-rays that did not reveal the cancer showed striking elevations of serum creatine kinase BB isoenzyme, which suggested the diagnosis and directed the subsequent evaluation. Creatine kinase BB isoenzyme elevation with widespread small cell carcinoma of the lung is probably more common than currently recognized, and is a useful diagnostic indicator for extensive stage small cell lung cancer, even in the absence of central nervous system metastases. PMID- 2988749 TI - Retroperitoneal germ cell tumors in childhood. A clinical and pathologic study of 11 cases. AB - During the last 54 years at the Children's Hospital 11 children (10 female, 1 male) had been treated for an extragonadal germ cell tumor arising in the retroperitoneum. There were eight teratomas (five mature, three immature), two endodermal sinus tumors and one tumor with a mixture of both components. Abdominal pain or discomfort was the usual presenting complaint, with the average age at diagnosis being 18 months. On physical examination, each child had a palpable abdominal mass usually localized in the upper quadrants. The finding of calcification, bone or teeth, on radiologic study was most helpful in establishing a preoperative diagnosis of teratoma. The preferred treatment for children with mature and immature teratomas is complete surgical resection; decisions regarding adjuvant therapy for patients with immature tumors must be evaluated on an individual basis. The prognosis remains guarded for children with unresectable primaries or those with endodermal sinus tumor. Of three tumor related deaths, two were due to metastatic endodermal sinus tumor and one resulted from extensive local growth by immature teratoma. Successful management of children with endodermal sinus tumor requires surgery aimed at debulking or complete resection in combination with aggressive adjuvant chemotherapy. PMID- 2988750 TI - Expression of neurofilament and neuron-specific enolase in small cell tumors of skin using immunohistochemistry. AB - Trabecular carcinoma of the skin (Merkel cell tumor), a neoplasm of putative neural origin, must be differentiated from other small cell tumors primary or metastatic to skin. In order to provide more objective diagnostic criteria, four were examined using monoclonal antibodies against neurofilament proteins (NF) and antiserum specific for neuron-specific enolase (NSE). Immunohistochemistry demonstrated immunoreactive NF in three and NSE in four cases. NF immunoreactivity was arranged in paranuclear balls, consistent with ultrastructural observations of aggregated intermediate filaments. A case of pulmonary oat cell carcinoma metastatic to the skin also contained immunoreactive NF and NSE. Although NF and NSE do not discriminate metastatic oat cell carcinoma from trabecular carcinoma of skin, they are useful antigens that provide objective criteria for recognizing tumors of neural histogenesis or neural differentiation. They should make it possible to exclude non-neural lesions from the differential diagnosis of small cell tumors of the skin. PMID- 2988751 TI - The value of the bone scan and bone marrow biopsy staging small cell lung cancer. AB - The charts of 112 patients with small cell lung cancer were reviewed in a retrospective fashion in order to define the role of the radionuclide bone scan and bone marrow biopsy in the staging of this disease. Both a radionuclide bone scan and bone marrow biopsy were performed on all patients at the time of diagnosis. Sixty-one percent of patients had a negative bone scan and negative biopsy; 22% had a positive bone scan and negative biopsy; 8% had a negative scan and positive biopsy; and 9% had a positive scan and positive biopsy. In 21 of the 44 patients with osseous involvement, no other focus of distant metastasis was found. The bone scan showed greater than or equal to 3 areas of increased uptake in 15 patients, 2 areas of increased uptake in 13 patients, and 1 area in 7 patients. The number of patients with bone marrow biopsy results positive for tumor in these 3 groups were 5, 3, and 2, respectively. Our study shows a lack of correlation between bone scan and bone marrow biopsy results. The bone scan and bone marrow biopsy identify independent patterns of osseous metastasis. Both procedures should be performed in the evaluation of patients with small cell lung cancer. PMID- 2988752 TI - Prognostic histologic features of resected small hepatocellular carcinoma (HCC) in Taiwan. A comparison with resected large HCC. AB - The morphologic features and their prognostic significance were analyzed in 83 surgically resected hepatocellular carcinomas (HCC): 44 of them were of less than or equal to 5 cm in diameter and 39 were larger. This study demonstrated a high prevalence of tumor capsule in both small and large HCCs, 86.4% and 84.6%, respectively. In small HCC, the capsule formation was significantly higher in the liver with cirrhosis (96.9%, or 31/32) than without (58.3%, or 7/12) (P less than 0.003). In both small and large HCCs, the most important histologic parameter influencing the tumor recurrence was the liver invasion. In the small but not in the large HCC, the invasion through the capsule also correlated well with recurrence (P less than 0.05). None of the 19 patients whose HCC were confined by a tumor capsule and did not invade the liver, had a recurrence during the 10 months or longer follow-up period. The significantly more favorable outcome of smaller HCC was related to the lower frequencies of liver invasion (P less than 0.001), portal vein involvement (P less than 0.01), and satellite formation (P less than 0.01). Tumor recurrence did not correlate with the tumor grading, presence of clear cells, liver cell dysplasia, or host inflammatory response. The HBsAg positivity in tumor cells was significantly higher in small (40.9%, or 18/44) than in large HCC (10.3%, or 4/39), suggestive of a gradual loss of the cytoplasmic expression of HBsAg in the tumor cells during the growth of HCC. PMID- 2988753 TI - Establishment and characterization of a human glioblastoma cell line with a stable karyotype and nullisomy 13. AB - A permanent cell line (HeRo) with a stable karyotype (80-84,XXYY) and with defined numerical and structural chromosome aberrations was established from a human glioblastoma, a highly malignant brain tumor. Transformation of these cells with SV40 led to a second permanent cell line (HeRo-SV) with a reduced, but also stable, karyotype (72-74,XXYY). The morphological appearance of the glioblastoma line was similar to the main component of the original tumor tissue. The transformed cells differed from their counterparts in accelerated growth, enhanced growth in soft agar, reduced growth conditions, expression of SV40 T antigen, and altered epitheloid morphology. Both cell lines have been grown in continuous culture for more than 2 years. The stability of both the biologic properties and the karyotypic changes induced by SV40 is quite remarkable. Both lines show a nullisomy 13. PMID- 2988755 TI - Preferential lymphocyte-mediated cytotoxicity of syngeneic target cells in chickens bearing tumors induced by avian sarcoma virus. AB - Splenic lymphocytes from chickens bearing tumors induced by avian sarcoma virus are able to cause the specific killing of cultured avian sarcoma cells. This cytotoxicity appears to follow classical patterns of syngeneic restriction. Little or no specific killing of tumor targets occurred when spleen cells from one inbred line of chickens were tested against allogeneic targets, although syngeneic killing proceeded relatively efficiently. Other patterns of immune reactivity did not appear to be syngeneically restricted. Namely, sera from tumor bearing hosts were equally reactive in indirect immunofluorescence assays with syngeneic and allogeneic target cells. And, peripheral blood lymphocytes from sensitized hosts could be stimulated equally well by tumor cell culture fluids of allogeneic or syngeneic origin. PMID- 2988754 TI - Abnormalities of chromosome 14 at band 14q11 in Japanese patients with adult T cell leukemia. PMID- 2988756 TI - Inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced nitroblue tetrazolium reduction in mouse peritoneal macrophages by various tumor promotion inhibitors. AB - The effects of various inhibitors on 12-O-tetradecanoylphorbol-13-acetate (TPA) induced reduction of nitroblue tetrazolium (NBT) in mouse peritoneal macrophages were investigated. The reduction was inhibited by phospholipase A2 inhibitors, such as dibromoacetophenone, the lipoxygenase inhibitor nordihydroguaiaretic acid, an NADH-dehydrogenase inhibitor, the microfilament inhibitor cytochalasin B, oxygen radical scavengers such as superoxide dismutase, antioxidants such as butyl hydroxyanisole and non-specific inhibitors such as retinoic acid. The reduction was not affected by the cyclooxygenase inhibitor indomethacin or the H2O2 scavenger catalase. PMID- 2988757 TI - Effects of altered Ca2+ and Mg2+ concentrations on proliferation and functional differentiation of the clonal insulin-producing cells RINm5F. AB - Effects of calcium and magnesium on proliferation and functional differentiation were investigated in the clonal insulin-producing rat cell line RINm5F. Variations of the extracellular concentrations of Ca2+ and Mg2+ had only minor effects on the cellular contents of these elements. Even when the medium was depleted of extracellular Ca2+ by the addition of EGTA, the Ca/Mg ratio corresponded to approximately 0.1. In a Mg2+-deficient medium a lowering of the extracellular calcium to 0.13 mM reduced the amount of insulin in the media but increased that in the cells. Irrespective of the concentration of Mg2+, depletion of Ca2+ was associated with low media levels of insulin and a reduced proliferation rate. In addition, the cells became smaller in size and showed an increased ability to exclude trypan blue. a rise of the magnesium concentration to 6.30 mM was accompanied by a decrease of the cellular content of insulin despite reduced amounts of hormone in the medium. This finding might be due to suppression of insulin biosynthesis, suggesting that the RINm5F cells have a markedly different metabolism and/or sensitivity to extracellular magnesium than non-tumorigenic beta-cells. PMID- 2988758 TI - Adriamycin does not affect the repair of X-ray induced DNA single strand breaks. AB - The ability of the antitumor antibiotic adriamycin (Ad) to inhibit the rejoining of DNA single strand breaks produced by X-rays was investigated in cultured cells. Chinese hamster ovary (CHO) cells were given 400 rad and were allowed to repair in the presence or absence of Ad for 60 min at 37 degrees C. The drug did not affect the ability of cells to repair DNA breaks and residual breaks found after the repair period were attributable to those induced by Ad alone. PMID- 2988759 TI - Superoxide dismutase levels in Chinese hamster ovary cells and ovarian carcinoma cells after hyperthermia or exposure to cycloheximide. AB - Superoxide dismutase (SOD) activity in Chinese hamster ovary (CHO) and ovarian carcinoma (OvCa) cells was measured after exposure to hyperthermia and correlated with the development of thermotolerance. The SOD activity of each cell type was largely copper- and zinc-containing SOD activity. Both cell types had similar but low levels of SOD activity when the cells were grown at 37 degrees C. After exposure for 2 h at 41.5 degrees C, SOD activity of OvCa cells, but not of CHO cells, was increased. After exposure to 45 degrees C for 15 min, SOD activity was also increased in the OvCa cells, but not in CHO cells. After 15 min at 45 degrees C followed by 1 h incubation at 37 degrees C, SOD activity was increased in OvCa and CHO cells; after an 8-h incubation at 37 degrees C, SOD activity doubled in each cell type. Thermotolerance is maximal after 2 to 3 h of exposure at 41.5 degrees C and after 8 to 10 h incubation at 37 degrees C following exposure to 15 min at 45 degrees C. The turnover of SOD activity in OvCa cells was estimated by the rate at which activity was lost following addition of cycloheximide (10 micrograms/ml). Twenty-four % of the activity was lost with a half-life of 10 min, and 76% was lost with a half-life of 4.5 h. Despite restriction of general protein synthesis 3 to 4 h after 45 degrees C hyperthermia, SOD activity was increased at 1 and 8 h after exposure, presumably coincidental with heat shock protein synthesis and development of thermotolerance. These data suggest that SOD activity may be important in protecting cells exposed to heat and that it may play a role in the development of thermotolerance. PMID- 2988760 TI - Distinct differentiation-inducing activities of gamma-interferon and cytokine factors acting on the human promyelocytic leukemia cell line HL-60. AB - The human promyelocytic leukemia cell line HL-60 and monoblastic leukemia cell line U937 undergo differentiation when induced by lymphokine and cytokine preparations. Growth inhibition, acquisition of immunoglobulin Fc receptors, increased expression of monocyte-related surface antigens, and an increase in lysosomal enzyme contents accompany maturation induced by gamma-interferon and other cytokine factors tested. Additionally, increased receptors for chemotactic peptide (fMLPR), increased hydrogen peroxide release in response to phorbol myristic acetate stimulation, and the release of prostaglandins (PGE2 and 6-keto PGF1a) follow exposure to lymphokine and cell line sources of myeloid colony stimulating activity (CSA). Gamma-Interferon (gamma-IFN) induced fMLPR in HL-60 (only at 1000 units/ml) but not in U937. Additionally, gamma-IFN did not induce prostaglandin release in either cell line. These myeloid colony-stimulating activity-associated differentiation-inducing factors were obtained from the human hepatoma++ cell line SK-Hep and bladder carcinoma cell line 5637, which were free of interferon activity. The 2-day phytohemagglutinin-induced lymphokine contained no detectable CSA and was a good source of differentiation activity. A simple, rapid assay for a new human CSA with pluripotent hematopoietic stimulating activity (pluripoietin) is described based on stimulation of [3H]glucosamine incorporation. Cell line conditioned media containing pluripoietin, purified pluripoietin, and gamma-IFN are active in this assay. These myeloid leukemia cell line differentiation factors are thus different from interferon and conventional CSA. These results suggest that endogenous human cytokines may have a role in the differentiation of leukemic as well as normal myeloid cells. PMID- 2988761 TI - Suppression of tumor promoter-induced chemiluminescence in mouse epidermal cells by several inhibitors of arachidonic acid metabolism. AB - The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) is a stimulator of chemiluminescence (CL) in SENCAR mouse epidermal cells. The CL response is TPA dose dependent (8 to 800 nM) as well as proportional to the number of cells used. Treatment with 166 nM TPA results in a CL response that peaks by 15 min although a strong response persists for over 30 min. The CL response can be inhibited by superoxide dismutase and the superoxide dismutase mimetic copper(II) (3,4 diisopropylsalicylic acid)2, suggesting that the CL response may be due to or mediated by superoxide anions. Catalase, which is specific for H2O2, and mannitol, which is a scavenger for hydroxyl radicals, had negligible inhibitory effects. The CL response is also inhibited by retinoic acid and the analogue ethyl all-trans-9-(4-methoxy-2,3,6-trimethylphenyl)-3,7-dimethyl-2,4,6,8- nonatetraenoate. A series of phorbol esters with different promoting abilities produced corresponding CL responses. The second stage tumor promoter mezerein is as effective as TPA in stimulating CL. Inhibitors of various parts of the arachidonic acid cascade were found to affect the TPA-induced CL response in a manner that corresponds to their effects in vivo tumor promotion experiments: agents which are predominantly lipoxygenase inhibitors, i.e., nordihydroguaiaretic acid, benoxaprofen, or agents which are effective against both lipoxygenase or cyclooxygenase, i.e., 5,8,11,14-eicosatetraynoic acid and phenidone, are effective in diminishing the CL response. Cyclooxygenase inhibitors, i.e., indomethacin and flurbiprofen, have no or a slight enhancing effect at low doses. These data suggest that at least a major part of the TPA induced CL response is due to the metabolism of arachidonic acid, most probably by the lipoxygenase(s). This CL assay may provide a useful system for studying the involvement of oxidants in tumor promotion. PMID- 2988763 TI - Characterization of the blood lymphocyte population in cattle infected with the bovine leukemia virus. AB - Blood leukocytes of cattle characterized in terms of bovine leukemia virus (BLV) infection and persistent lymphocytosis (PL) were examined for the presence of lymphocyte subpopulation markers and viral antigens. The percentages of cells with surface and intracytoplasmic immunoglobulin M (IgM) and erythrocyte-antibody rosetting cells agreed closely in all infected cattle. This correlation and the results of double labeling experiments indicate that virtually all the surface IgM-positive B-lymphocytes in the blood of these animals carry Fc receptors. In PL cattle, the percentages of surface IgM-positive cells were more than twice those of normal cells and accounted for all the increase in peripheral blood lymphocytes. B-cells accounted for most of the increase in peripheral blood lymphocytes seen in cattle with PL. In contrast, most BLV-infected, nonlymphocytotic cattle had normal percentages of B-cells. Thus, the expansion of the B-cell population in blood, while being a conspicuous characteristic of PL, is not necessarily a consequence of BLV infection per se. Comparisons of the percentages of IgM-positive and erythrocyte-antibody complement-rosetting cells, together with the results of double labeling experiments, indicate that about one half the B-cells in the blood of cattle with PL lacked C-3 receptors. The proportion of these cells (most likely immature B-lymphocytes) was smaller in the blood of BLV-infected nonlymphocytotic cattle. Direct comparison showed that, in BLV-infected cattle with or without PL, and in BLV-free cattle, virtually all erythrocyte-rosetting blood cells had peanut agglutinin receptors. With only one exception, the numbers of erythrocyte-positive cells in the blood of BLV-infected cattle with or without PL were within normal values. The "null" blood cell population, estimated as the difference between the IgM-positive and erythrocyte positive populations, was essentially unaffected in BLV-infected cattle without PL, but it was absent in PL cattle. The large majority of the B-lymphocytes present in the blood of cattle with PL were infected with BLV. The proportion of infected B-lymphocytes in the blood of BLV-positive nonlymphocytotic cattle was much lower. Even in cattle with low or moderate levels of BLV-infected blood lymphocytes, the percentages of these cells were remarkably constant during the 12-month period of the study. The data indicate that most of the BLV-infected B lymphocytes of cattle with PL lack C-3 receptors. PMID- 2988762 TI - Correlations between intercalator-induced DNA strand breaks and sister chromatid exchanges, mutations, and cytotoxicity in Chinese hamster cells. AB - Intercalator-induced DNA strand breaks in mammalian cells represent topoisomerase II:DNA complexes trapped by intercalators. These complexes are detected as protein-associated DNA single-strand breaks (SSB) and DNA double-strand breaks (DSB) by filter elution. Using Chinese hamster lung fibroblasts (V79 cells) that were treated for 30 min with various concentrations of 4'-(9 acridinylamino)methanesulfon-m-anisidide or 5-iminodaunorubicin, we measured DNA strand breaks (SSB and DSB), sister chromatid exchanges (SCE), mutations at the hypoxanthine:guanine phosphoribosyltransferase locus, and cell killing. Further, we correlated DNA strand breakage with the three other parameters. Both drugs induced SCE, mutations, and cell killing at concentrations which also produced reversible DNA strand breaks. While the quantity of DSB correlated with SCE, mutations, and cytotoxicity for both drugs, we found more SCE, mutations, and cytotoxicity per SSB in cells treated with 5-iminodaunorubicin than in those treated with 4'-(9-acridinylamino)methanesulfon-m-anisidide. These data show that the DSB (but not the SSB) induced by 4'-(9-acridinylamino)methanesulfon-m anisidide and 5-iminodaunorubicin at DNA topoisomerase II binding sites correlated closely with SCE, mutations, and cell killing and could therefore be responsible for their production. PMID- 2988764 TI - An amplified immunoperoxidase assay to detect bovine leukemia virus expression: development and comparison with other assays. AB - An amplified immunoperoxidase (AIP) assay using an avidin:biotin complex was developed to detect bovine leukemia virus (BLV) antigen expression in lymphocytes which had been cultured 24 h and fixed with acetone. Nonspecific reactions were eliminated by absorbing the test serum with 100% horse or cow serum. DNA synthesis inhibition did not decrease the number of AIP-positive cells, and there were no apparent preferential losses of major lymphocyte subpopulations during culture. Both viable and nonviable BLV-expressing cells were detected. Thus, the number of AIP-positive cells seems to be a good estimate of the minimum number of infected lymphocytes present in the uncultured blood cells. In direct comparisons, twice as many BLV-expressing cells were detected with the AIP assay as with an indirect immunofluorescence test. The AIP assay is as sensitive as the syncytia infectivity assay and only slightly less sensitive than an immunoperoxidase infectivity assay for detecting BLV-infected lymphocytes in the blood of infected cattle that were in early stages of infection and/or had low titers of antiviral antibodies. The AIP assay is the most sensitive, rapid, and reproducible procedure available for the identification of individual cells infected with BLV. This assay may be of great value in studies on the biology of BLV infection. PMID- 2988765 TI - Histological types of carcinoma of the uterine cervix and the detectability of human papillomavirus DNA. AB - Using the Southern DNA hybridization technique, tissues from 17 cases of invasive carcinoma of the uterine cervix, including nine cases of squamous cell carcinoma, four cases of adenocarcinoma, one case of adenosquamous carcinoma, and three cases of undifferentiated carcinoma, were examined for the presence of human papillomavirus (HPV) DNA. None of the studied cases had histologically confirmed association of condyloma acuminatum or cervical intraepithelial neoplasia in the vicinity. HPV DNA was detected in two of 17 cases under low stringency conditions. One lesion was undifferentiated carcinoma, and another was squamous cell carcinoma. Hybridization under high stringency conditions with a variety of HPV DNA probes indicated the presence of HPV-16 in these two lesions. The other HPV-positive lesion was adenocarcinoma, demonstrating weak hybridizations with HPV-2 and HPV-16 DNA probes only under high stringency conditions. Altogether, three of 17 cases (17.6%) contained HPV DNA. This observation contrasts to the rate of HPV DNA present in 15 of 18 cases (83.3%) of the tissues of cervical intraepithelial neoplasia. Our data suggest that HPV was not consistently detected in invasive squamous cell carcinoma, despite the frequent association of HPV with its supposed precursor lesions of cervical intraepithelial neoplasia. PMID- 2988766 TI - Adriamycin-enhanced membrane lipid peroxidation in isolated rat nuclei. AB - Isolated rat liver nuclei enzymatically activated Adriamycin to the electron spin resonance-detectable semiquinone free radical in the presence of reduced nicotinamide adenine dinucleotide phosphate (NADPH). This process resulted in the enhancement of oxyradical-mediated peroxidation of the nuclear envelope membrane unsaturated phospholipids, measured as malonaldehyde equivalents by the thiobarbituric acid method. Peroxidation required the inclusion of NADPH and catalytically active protein, presumably NADPH:cytochrome P-450 reductase, and was enhanced more than 5-fold by Adriamycin. It was observed that Adriamycin stimulated nuclear membrane peroxidation was diminished by superoxide dismutase, reduced glutathione, the hydroxyl radical scavenger, 1,3-dimethylurea, and by the metal ion chelator, diethylenetriaminepentaacetic acid, indicating that multiple species of reactive oxygen and trace amounts of metal ions (iron) were required in the peroxidation reaction. The generation of superoxide and hydroxyl radicals was confirmed by 5,5-dimethyl-1-pyrroline-N-oxide spin trap electron spin resonance spectroscopy. Calf thymus DNA added to incubations containing nuclei and NADPH caused a pronounced concentration-dependent inhibition of Adriamycin stimulated lipid peroxidation. It was found that nuclei incubated with Adriamycin (300 microM) accumulated 128 nmol of the drug per mg of nuclear protein within 1 h, apparently because the Adriamycin was internalized and bound to the DNA and nuclear protein. These results suggest that some of the toxic effects of Adriamycin observed in the nucleus could result, directly or indirectly, through the peroxidation of the unsaturated lipids of the nuclear membrane. PMID- 2988767 TI - Effects of vitamin A and dexamethasone on collagen degradation in mouse mammary adenocarcinoma. AB - Collagenases and other neutral proteases in tumors may facilitate tumor extension, invasion, and subsequent metastasis. We report the effects of vitamin A and dexamethasone, known inhibitors of collagenase production in vitro, on the collagen metabolism of mouse mammary adenocarcinoma and its capsule, borne by C3H/HeJ mice. The weight of the capsule was about 4% of the tumor, yet the total collagen content of the capsule was about 10-fold greater than that of the tumor tissue; tumor cells had no detectable collagen. With tumor growth, the collagenase and other neutral protease activities were increased in the tumor tissue; a negative correlation existed between collagenase activity and collagen content of the capsule. The protease activities of the tumor borne by vitamin A treated hosts were about 50% lower than those of the controls; this coincided with a slight increase in the collagen content of the capsule. In contrast, the collagen content of the capsule borne by dexamethasone-treated hosts was 50% less than that of the controls; the protease activities were similar to the controls and occurred with tumor invasion and metastasis. Results suggest that the collagen metabolism of the capsule may be an indicator of proteolytic events within the tumor and the metastatic potential of the tumor that, in turn, suggests the possibility of preventing metastasis by inhibiting the production of collagenases and other neutral proteases, thereby localizing the tumor cells within the capsule. Vitamin A could be used for that purpose. PMID- 2988768 TI - Isolation of retinoic acid-resistant clones from human breast cancer cell line MCF-7 with altered activity of cellular retinoic acid-binding protein. AB - After ethyl methane sulfonate mutagenesis of the mammary carcinoma cell line, MCF 7, we have isolated three clones, U-2, U-3, and U-9, resistant to retinoic acid. These three clones showed more than a 1000-fold higher level of resistance to retinoic acid than the parental MCF-7 cells when assayed by colony formation in monolayer culture system or by growth curves. The three resistant clones showed a 200-fold higher resistance to 13-cis-retinoic acid, about 10-fold higher resistance to retinol, and about 2-fold higher resistance to retinyl acetate, respectively, than MCF-7. Binding of [3H]retinoic acid or [3H]-retinol to a cellular fraction in situ showed apparent decrease of the specific binding of retinoic acid in U-2, but there was no such specific fraction bound to retinol in U-2 and MCF-7. Sucrose gradient analysis with cytoplasmic fraction showed little, if any, cellular retinoic acid-binding protein in U-2, but a significant amount of the cellular retinoic acid-binding protein could be found in MCF-7. By contrast, there was no activity of cellular retinol-binding protein in both MCF-7 and U-2. The sensitivity or resistance of mammalian cells in culture to retinoic acid is discussed in relation with cellular binding activity for vitamin A. PMID- 2988769 TI - Limited impact of total parenteral nutrition on nutritional status during treatment for small cell lung cancer. AB - During a randomized trial of total parenteral nutrition (TPN) in patients with small cell lung cancer, we evaluated the short- and long-term effects of 4 weeks of TPN on nutritional assessment parameters. All 119 patients who were accrued to the study received the same chemotherapy and radiotherapy protocol which extended over a 1-year period: 57 patients received TPN; and 62 served as controls. At base line, patients with greater than 5% pretreatment weight loss had significantly lower levels of serum albumin, total iron-binding capacity, and creatinine/height index. TPN administration led to a significant increase in mean caloric intake and weight compared with controls (P less than 0.0001). In the short-term study, body fat, as measured by triceps skinfold thickness, was maintained, and there was a small increase in arm muscle circumference. Serum albumin and hematocrit decreased but promptly returned to pretreatment levels when TPN was stopped. There were no long-term differences in any of the nutritional assessment parameters between the two groups. PMID- 2988770 TI - Antigenicity of human T-cell leukemia virus-associated gp52: greater response in leukemia patients compared to healthy donors exposed to the virus. AB - Monoclonal antibody HT462 recognizes a human T-cell leukemia virus type I (HTLV I)-associated Mr 52,000 glycoprotein (HA-gp52), which is found on the surface of HTLV-infected cells. Whether HA-gp52 is encoded by the virus or by the infected cells has not yet been established. Using monoclonal HT462 in a competitive binding assay, natural human antibodies specific for HA-gp52 were detected in 97% of the patients from the United States, the Caribbean, and Japan with adult T cell leukemia but not in healthy donors not exposed to HTLV-I. In contrast, antibodies to HA-gp52 occur in healthy virus-exposed donors, but at a lower prevalence than that observed in patients. Among Japanese from HTLV-I-endemic areas and exposed to the virus as indicated by the presence of antibodies to disrupted HTLV-I, 93% of adult T-cell leukemia patients were also seropositive for HA-gp52 compared to only 16% of healthy individuals. Differing sensitivities in the methods of assaying antibodies to HTLV and HA-gp52 were not responsible for these observations as shown by the lack of correlation of HTLV-I antibody titer with the presence of antibody to HA-gp52. Among adult T-cell leukemia patients, antibody titers to HTLV-I and HA-gp52 also varied independently. These results indicate that HA-gp52 in humans is antigenic and correlated with disease. Detection of antibody to this protein in asymptomatic individuals may be indicative of a predisease condition. PMID- 2988771 TI - Lipid cerebral embolization following lymphogram. PMID- 2988772 TI - The pharmacology of myoclonus. PMID- 2988773 TI - Cisplatin and etoposide combination chemotherapy for refractory small cell carcinoma of the lung. AB - Twenty-nine patients with refractory recurrent small cell carcinoma of the lung were treated with cisplatin (40 mg/m2) and etoposide (200 mg/m2) each day for 3 days, repeated every 3-4 weeks. Fifteen of these patients had received etoposide in their original treatment regimen. Fifteen (52%) of all patients had a major response, as did nine (60%) of the patients with prior exposure to etoposide. Myelotoxicity was moderately severe. The median duration of responses was 3 months (range, 6-36 weeks). This study suggests synergism between cisplatin and etoposide. The toxicity seen in this heavily pretreated group of patients suggests that smaller doses be studied in this group. The synergism may be best utilized in the initial regimens against small cell lung cancer. PMID- 2988774 TI - Eastern Cooperative Oncology Group phase II studies in advanced measurable colorectal cancer. I. Razoxane, Yoshi-864, piperazinedione, and lomustine. AB - During a 6-month interval, Eastern Cooperative Oncology Group randomized 127 patients who had received prior chemotherapy, and who had advanced measurable, surgically incurable colorectal cancer, to receive piperazinedione (PZD), Yoshi 864, or razoxane (ICRF-159). The observed response (and median survival) rates were: PZD, one of 35 patients (17 weeks); Yoshi-864, one of 34 (19 weeks), and ICRF-159, none of 38 (23 weeks). Among 107 evaluable patients, there were five episodes of life-threatening toxicity with PZD (one death) and four with ICRF-159 (two deaths). In the same protocol, 42 evaluable patients who had not received prior chemotherapy were randomized to be treated with lomustine (CCNU) or one of the three drugs in the "previously treated" trial. One CR (41 weeks) was seen with ICRF-159 and two PRs were seen with CCNU. Life-threatening toxicity occurred in three patients, two who received CCNU (one death) and one who received PZD. No survival advantage was seen. We do not encourage further phase II trials in colorectal cancer with the agents studied. PMID- 2988775 TI - Treatment of small cell lung cancer with 6-mercaptopurine: a phase II study. PMID- 2988776 TI - Alpha-adrenoceptor stimulation, lysophosphoglycerides, and lipid peroxidation in reoxygenation induced calcium uptake in rabbit myocardium. AB - Calcium uptake on reoxygenation of hypoxic cardiac muscle is well documented. Alpha-adrenoceptor stimulation by released catecholamines, lysophosphoglycerides and lipid peroxidation have all been suggested as mediators of this effect. We have measured the uptake of Ca2+ on reoxygenation in the isolated arterially perfused interventricular septum of the rabbit heart. The alpha agonist phenylephrine (1 mumol X litre-1) did not alter calcium uptake, and the presence of either prazosin (1 mumol X litre-1) or phentolamine (10 mumol X litre-1) did not alter the reoxygenation induced Ca2+ uptake. Lysophosphatidylcholine caused an increase in Ca2+ uptake above 8 mumol X litre-1 but also produced a simultaneous increase in the distribution volume of 51Cr-EDTA, an extracellular space marker, indicating loss of membrane integrity. Hydrogen peroxide and cumene hydroperoxide both caused an increased Ca2+ uptake, but no disruption of the cell membrane; the effect on Ca2+ uptake could be inhibited by Ni2+ ions. Alpha adrenergic stimulation and lysophosphoglycerides do not appear to be key to Ca2+ uptake on reoxygenation, but lipid peroxidation of the sarcolemma is a possible mechanism. PMID- 2988777 TI - [Quantification of left-right intracardiac shunts using radioisotopes]. PMID- 2988779 TI - Hepatocellular carcinoma presenting as a fever of undetermined origin. PMID- 2988778 TI - Study of the properties of MgATP2--induced stationary bends in demembranated sea urchin sperm. AB - Methods of demembranation and reactivation of Lytechinus pictus sperm were developed that result in non-motile sperm which take on a stable bend of about 3.5 radians at the proximal end of the cell. The middle and distal portion of the flagellum is relatively straight or slightly bent in the same direction forming a somewhat "C" shaped sperm cell. In these studies, we refer to this characteristic shape as the quiescent form, and as opposed to "rigor wave" sperm, the quiescent form is induced and maintained by a relatively high concentration of MgATP2- (greater than 0.2 mM). Other conditions important to the production and maintenance of the quiescent form in demembranated sperm include: starting with concentrated, undiluted sperm, maintaining low Ca++ in the demembranation buffer, using a minimum of 0.2 mM MgATP2- and pH of 7.9-8.1 in the reactivation buffer. Deviation from some of these conditions results in a dramatic increase in motile, asymmetrically beating sperm. Addition of 0.4 mM CaCl2 to the reactivation buffer increased the proximal bend angle to 5 radians. The induction and maintenance of the stationary bend is mediated by dynein activity: "rigor wave" sperm were transformed to the quiescent form upon 0.2 mM ATP addition; micromolar vanadate abolished the quiescent form by "relaxation" of the proximal bend; and the vanadate relaxed sperm were restored to quiescent form by catechol. Importantly, 20 microM cAMP activated motility of the otherwise quiescent-form sperm. Quiescent-form, demembranated sperm were also activated by mild trypsin digestion. These and other data suggest that the quiescent-form sperm are trapped at the end of the principal bend, and these data are consistent with the proposal that the single stationary bend results from asymmetry of active microtubule sliding [Gibbons and Gibbons, (1980): J. Cell Biol. 84:13-27]. PMID- 2988780 TI - Cyclic-AMP-dependent protein kinase type II is associated with the Golgi complex and with centrosomes. AB - The subcellular distribution of the type II enzyme of cAMP-dependent protein kinase (cAMP-dPK II) in epithelial and fibroblastic cells was determined by indirect immunofluorescence microscopy. In interphase cells both regulatory (R II) and catalytic (C) subunits were concentrated in a perinuclear area. By comparison of the R II distribution with the location of a bona fide Golgi membrane constituent, this area was identified as the Golgi complex. The cytochemical localization of R II was confirmed by subcellular fractionation. In addition, cAMP-dPK II was associated with microtubule-organizing centers, in particular with mitotic spindle poles. These distributions of cAMP-dPK II probably represent important factors in mediating the effects of cAMP on basic cellular activities ranging from secretion and proliferation to cell shape and motility. PMID- 2988781 TI - Site-specific inversion: enhancers, recombination proteins, and mechanism. PMID- 2988782 TI - Malignant transformation of a growth factor-dependent myeloid cell line by Abelson virus without evidence of an autocrine mechanism. AB - Abelson virus has been used to transform cells of a murine, factor-dependent myeloid cell line (FD). Factor-independent (FI) cell lines were derived, which expressed the viral genome and were tumorigenic in syngeneic mice. Karyotypic analysis of FI cells before and after passage in vivo indicated that the tumorigenic cells were derived from FD cells. Northern gel analysis of mRNA, bioassay of culture supernatants, and the density-independent growth of the FI cells indicated that the transformation had not induced the synthesis of the hemopoietic growth factors normally required to support the FD cells, that is, granulocyte-macrophage CSF or Multi-CSF. The FD and FI cells displayed similar numbers of cell surface receptors for Multi-CSF (IL-3) and GM-CSF. We conclude that Abelson virus transformation of this line from factor-dependence to factor independence and tumorigenicity did not involve autocrine stimulation. PMID- 2988783 TI - Neoplastic transformation of mast cells by Abelson-MuLV: abrogation of IL-3 dependence by a nonautocrine mechanism. AB - Normal mast cells can be propagated in culture when medium is supplemented with interleukin-3 (IL-3). We demonstrate that Abelson-MuLV (Ab-MuLV) infection of mast cells eliminates dependence on IL-3 for growth. By contrast, Harvey, BALB, and Moloney MSV, which also productively infect mast cells, are unable to relieve IL-3 dependence. Ab-MuLV-induced IL-3-independent lines express the v-abl specific transforming protein and have phenotypic characteristics of mast cells. These cells also possess high cloning efficiencies in soft agarose and are tumorigenic in nude mice. In addition, Ab-MuLV induces transplantable mastocytomas in pristane-primed adult mice resistant to lymphoid transformation, defining a new hematopoietic target for malignant transformation by this virus. None of the Ab-MuLV-derived transformants express or secrete detectable levels of IL-3 nor is their growth inhibited by anti-IL-3 serum. These results argue that Ab-MuLV abrogation of the IL-3 requirement is not due to an autocrine mechanism. PMID- 2988784 TI - c-erbB activation in ALV-induced erythroblastosis: novel RNA processing and promoter insertion result in expression of an amino-truncated EGF receptor. AB - ALV-induced erythroblastosis results from the specific interruption of the host oncogene, c-erbB, by the insertion of an intact provirus. Integrated proviruses are oriented in the same transcriptional direction as c-erbB, and expression of truncated c-erbB transcripts is observed. Evidence, including sequence analysis of cDNA clones, indicates that transcription of truncated c-erbB mRNA is initiated in the 5' LTR of the integrated provirus. This transcript is processed through a series of remarkable splicing reactions to yield viral gag and env sequences fused to erbB sequences. These results establish a novel pathway of promoter insertion oncogenesis that stands in contrast to the pathways used in the activation of c-myc in B lymphomas. PMID- 2988785 TI - Sites of allosteric shift in the structure of the cyclic AMP receptor protein. AB - We have characterized crp mutations in E. coli that allow CRP to function without cAMP. crp* mutants carrying a deletion of the gene encoding adenylate cyclase (cya) show significant lac expression. Cyclic GMP, normally an ineffective activator of CRP+, can stimulate these mutant CRP*s to permit greater lac expression in vivo. Cyclic AMP binding to the amino-terminal domain of CRP+ induces an allosteric transition that changes the DNA-binding property of the carboxy domain. The CRP* phenotype is caused by substitution of amino acids with bulkier side chains in the D alpha-helix of the protein's carboxy domain, near the hinge connecting the two domains. These results are consistent with a model in which the mutant CRP*s assume, in part, a conformation normally evoked only by cAMP binding: one in which the relative orientation of the C, D, and F alpha helices is altered. We define precisely the amino acids of these alpha-helices that interact to cause the allosteric shift. PMID- 2988786 TI - Positive control of a regulon for defenses against oxidative stress and some heat shock proteins in Salmonella typhimurium. AB - S. typhimurium become resistant to killing by hydrogen peroxide and other oxidants when pretreated with nonlethal levels of hydrogen peroxide. During adaptation to hydrogen peroxide, 30 proteins are induced. Nine are constitutively overexpressed in dominant hydrogen peroxide-resistant oxyR mutants. Mutant oxyR1 is resistant to a variety of oxidizing agents and overexpresses at least five enzyme activities involved in defenses against oxidative damage. Deletions of oxyR are recessive and uninducible by hydrogen peroxide for the nine proteins overexpressed in oxyR1, demonstrating that oxyR is a positive regulatory element. The oxyR1 mutant is also more resistant than the wild-type parent to killing by heat, and it constitutively overexpresses three heat-shock proteins. The oxyR regulatory network is a previously uncharacterized global regulatory system in enteric bacteria. PMID- 2988787 TI - Hin-mediated site-specific recombination requires two 26 bp recombination sites and a 60 bp recombinational enhancer. AB - The alternate expression of flagellin genes in Salmonella is the result of an inversion of a 996 bp segment of chromosomal DNA. We have analyzed the components of this site-specific recombination reaction in an in vitro system derived from E. coli. Efficient Hin-mediated inversion requires the 20,000 MW Hin protein and a proteinase K-sensitive host component. The supercoiled DNA substrate must contain two 26 bp recombination sites in inverted configuration and a 60 bp sequence that increases the rate of recombination over 20-fold. This recombinational enhancer can function at many different locations and consists of at least two noncontiguous sequence domains whose relative orientation, but not precise spacing, with respect to each other is important. Synthetically derived wild-type and mutant recombination sites were constructed to analyze the sequence and structural features that are important within the recombination site. PMID- 2988788 TI - Signals for site-specific cleavage of HSV DNA: maturation involves two separate cleavage events at sites distal to the recognition sequences. AB - Mature Herpes Simplex Virus (HSV) genomes are cleaved from concatemeric precursors by a site-specific mechanism. These cleavage events are probably coupled to the encapsidation process. Sequences within the terminal repeat of HSV DNA are necessary for the cleavage and packaging reactions, and are also thought to be responsible for high frequency genome isomerization events. Here we present evidence to show that two viral DNA cleavage and packaging signals reside within a 250 bp subfragment of the terminal repeat, that the termini of mature viral DNA are generated by a process involving two separate DNA cleavages at sites distal to the cleavage signals, and that the sequences between these two cleavage sites are duplicated by the DNA maturation system. PMID- 2988789 TI - Isomerization of herpes simplex virus 1 genome: identification of the cis-acting and recombination sites within the domain of the a sequence. AB - Previous studies have shown that the a sequence located at the termini and at the junction between the L and S components is the site-specific, cis-acting sequence mediating the inversions of herpes simplex virus 1 DNA. We constructed mutated a sequences, inserted them into the thymidine kinase gene, and recombined them into the L component of the viral genome. Deletion of Uc or Ub domains of the a sequence did not affect inversions, whereas the deletion of direct repeat #4 (DR4) drastically reduced their frequency. Deletion of both direct repeat #2 (DR2) and DR4 abolished inversions. Recombinational events leading to inversions appear to occur through DR2, and possibly DR4. These results complement previous studies showing that most of one DR1 sequence can also be dispensed with and are consistent with the hypothesis that DR4 and possibly DR2 are the cis-acting sites for the inversions mediated by the a sequence. PMID- 2988790 TI - The location of cis-acting regulatory sequences in the human T cell lymphotropic virus type III (HTLV-III/LAV) long terminal repeat. AB - The location of cis-acting regulatory sequences within the long terminal repeat (LTR) of the human T cell lymphotropic virus type III (HTLV-III/LAV) was determined. An enhancer element capable of increasing the rate of transcription from a heterologous promoter, irrespective of distance and orientation, is located between nucleotides -137 and -17 (cap site +1). The promoter sequences present near the TATA box respond to heterologous enhancers. The sequences present between nucleotides -17 and +80 are responsive to HTLV-III-associated trans-acting regulatory factors. Activation of these sequences by the viral regulatory factors requires the presence of a functional enhancer. The enhancer requirement is nonspecific, as the enhancer sequences of RSV, HTLV-I, and SV40 can functionally replace the HTLV-III enhancer. These findings define a new type of regulatory element, provide insight into the mechanisms that regulate HTLV-III gene expression, and may help to explain the effects of this virus on infected cells. PMID- 2988791 TI - Two simultaneously active VSG gene transcription units in a single Trypanosoma brucei variant. AB - Trypanosomes can change their surface coat either by slotting a different surface antigen gene copy into an active (telomeric) expression site or by activating a new VSG gene expression site and inactivating the old one. How expression sites are activated or inactivated is not clear. We report an exceptional trypanosome variant in which the inactivation of a surface antigen gene is accompanied by a 30 kb DNA insertion 5' of the gene. Transcription of the region upstream of the insertion continues unaltered and retains the characteristic insensitivity to alpha-amanitin of VSG gene transcription units, showing that the expression site is still active. The expressed VSG gene in this trypanosome variant resides in another telomere. Hence, two VSG gene transcription units can be simultaneously active. This argues against a single mobile activating element controlling VSG gene transcription and favors a stochastic model of telomere activation/inactivation. PMID- 2988792 TI - A truncated form of the bacteriophage Mu B protein promotes conservative integration, but not replicative transposition, of Mu DNA. AB - The phage-encoded proteins required for conservative integration of infecting bacteriophage Mu DNA were investigated. Our findings show that functional gpA, an essential component of the phage transposition system, is required for integration. The Mu B protein, which greatly enhances replicative transposition of Mu DNA, is also required. Furthermore, a truncated form of gpB lacking 18 amino acids from the carboxy terminus is blocked in replicative transposition, but not conservative integration. Our results point to a more prominent role for gpB than simply a replication enhancer in Mu DNA transposition. The ability of a truncated form of B to function in conservative integration, but not replicative transposition, also suggests a key role for the carboxy-terminal domain of the protein in the replicative reaction. The existence of a shortened form of gpB, which uncouples conservative integration from replicative transposition, should be invaluable for future dissection of Mu DNA transposition. PMID- 2988794 TI - DNA gyrase and its complexes with DNA: direct observation by electron microscopy. AB - Electron microscopy of DNA gyrase holoenzyme, of gyrase A subunits, and of the complexes of both species with DNA enables us to deduce the relative locations of subunits in the holoenzyme and to indicate a plausible path for DNA complexed with gyrase. The structural results are discussed in terms of certain models for directional DNA strand transport. PMID- 2988793 TI - Mechanism of transposition of bacteriophage Mu: structure of a transposition intermediate. AB - Mu transposition works efficiently in vitro and generates both cointegrate and simple insert products. We have examined the reaction products obtained under modified in vitro reaction conditions that do not permit efficient initiation of DNA replication. The major product is precisely the intermediate structure predicted from one of the current models of DNA transposition. Both cointegrates and simple inserts can be made in vitro using this intermediate as the DNA substrate, demonstrating that it is indeed a true transposition intermediate. The requirements for efficient formation of the intermediate include the Mu A protein, the Mu B protein, an unknown number of E. coli host proteins, ATP, and divalent cation. Only E. coli host proteins are required for conversion of the intermediate to cointegrate or simple insert products. Structures resulting from DNA strand transfer at only one end of the transposon are not observed, suggesting that the strand transfers at each end of the transposon are tightly coupled. PMID- 2988796 TI - The effect of 4-aminopyridine-induced increased neuromuscular activity on the metabolism of developing muscles in chick embryos. AB - Chick embryos were treated with 4-aminopyridine (4 X 100 micrograms) during a critical stage of muscle development, and the effect of enhanced neuromuscular activity upon energy metabolism was studied in two fast-twitch muscles and a slow tonic muscle. In the slow-tonic muscles of treated embryos, the specific activities of creatine kinase (CK) and lactate dehydrogenase (LDH) were reduced by 11 and 21%, respectively, compared with control values, whereas the ratios of the CK-MB isoforms and the LDH-H subunits increased to 125 and 135% of the control values, respectively. No significant changes could be shown in the enzymatic pattern of fast muscles. These results indicate that a moderate increase in neuromuscular activity of the chick embryo primarily influences the metabolism of developing slow muscles, promoting the development of an enzyme profile characteristic of slow oxidative fibres. PMID- 2988795 TI - HTLV-III env gene products synthesized in E. coli are recognized by antibodies present in the sera of AIDS patients. AB - The envelope gene of HTLV-III, the retrovirus directly linked to AIDS, encodes a protein of 856 amino acids. Our sequence analysis of the cloned HTLV-III (HXB-3) env gene and its comparison with other isolates reveal significant divergence, especially in the external portion of this protein. A large segment of the env gene (1800 bp) was inserted into the expression vector pEV-vrf3, and a corresponding 68 kd protein, which encompasses both the extracellular and the membrane-associated regions of the native protein, was produced in E. coli. Several smaller polypeptides, which appear to be internal initiation products, were also produced. All 50 AIDS patient sera obtained from different locations in the United States specifically recognized the bacterially synthesized envelope proteins, as judged by Western blots. This suggests that these proteins will be useful for the diagnosis of HTLV-III infection and possibly as a vaccine against AIDS. PMID- 2988797 TI - Change of hyaluronic acid synthesis during differentiation of myogenic cells and its relation to transformation of myoblasts by Rous sarcoma virus. AB - Hyaluronic acid synthesis was examined in cultures of differentiating chick embryo muscle cells before, during and after fusion. Prior to fusion, hyaluronic acid was synthesized and secreted into the medium, but once fusion began this synthesis was reduced significantly. Synthesis then increased again after completion of fusion. Thus, production of hyaluronic acid was lowest at the time of or right before cell fusion. When myoblasts were transformed by Rous sarcoma virus (RSV), a higher amount of hyaluronic acid was synthesized, and cells were not able to fuse. The turnover rate of hyaluronic acid might be different between myotubes and RSV-transformed myoblasts. The addition of exogenous hyaluronic acid to myoblast cultures resulted in the partial inhibition of fusion. The effect was reversible because fusion took place after removal of the exogenous hyaluronic acid. These observations suggest that hyaluronic acid plays an important role in the differentiation of myogenic cells, and that elevated hyaluronic acid synthesis may partly be the reason for inhibition of myotube formation upon transformation by Rous sarcoma virus. PMID- 2988798 TI - Asymmetry in the recognition of HLA-A3 molecules by virus-specific cytotoxic T cells. AB - Cytotoxic T cells specific for influenza virus A/HK or Epstein-Barr virus were used to study the heterogeneity of the HLA-A3 molecule. Variability of the recognition of HLA-A3 in both systems was observed. The hierarchy was both effector cell and target cell specific. An extreme example of the hierarchy of HLA-A3 recognition is the following. Virus-specific cytotoxic T lymphocytes of a given donor were found to recognize all HLA-A3-matched target cells, including target cells of a donor from whom the virus-specific effector cells did not recognize target cells of that given donor: Donor A recognizes target B but donor B does not recognize target A. Both will recognize a third HLA-A3-matched target cell C. Cold target inhibition studies confirmed that the recognition of target cell B by effector cell A involved the recognition of only HLA-A3. Examples of such asymmetric recognition were found in both influenza A and Epstein-Barr virus specific cytotoxic T-lymphocyte responses but not one combination was asymmetric in both systems. This suggests that influenza virus A/HK-specific cytotoxic T lymphocytes recognize other HLA-A3 histotopes than do Epstein-Barr virus-specific cytotoxic T lymphocytes. PMID- 2988799 TI - Alveolar ridge augmentation with hydroxylapatite: a report on patient satisfaction. PMID- 2988800 TI - Tumor-promoting, phorbol ester-induced phosphorylation of cell-surface transferrin receptors in human erythroleukemia cells. AB - When human erythroleukemia cells (K562) were exposed to phorbol-12-myristate 13 acetate (PMA), phosphorylation of transferrin receptors was enhanced 5-fold with 10(-7) M PMA and 7-fold with 10(-6) M PMA, but not with 4 alpha-phorbol (5 X 10( 7) M). Stimulation took place in serine residues in the cytoplasmic domain of the receptor. Although phosphorylation in the control cells took place in both cell surface and intracellular receptors, phosphorylation in PMA-treated cells increased only in the cell-surface receptors, not in the intracellular receptors. The number of receptors on the cell surface increased slightly with the increase in phosphorylation at the cell surface, in the PMA-treated cells. No difference in transferrin binding was found for the control and PMA-treated cells. These results indicate that enhanced phosphorylation of the transferrin receptor takes place on the cell surface only and that it presumably is mediated by protein kinase C. PMID- 2988802 TI - Inhibitors of angiotensin-converting enzyme in crude drugs. II. PMID- 2988801 TI - [Human papillomavirus disease of the cervix uteri]. PMID- 2988803 TI - Pharmacological activities in the mouse of delta 9-tetrahydrocannabinol metabolites oxidized at the 8-position. PMID- 2988804 TI - Thermotropic transitions of lipids in porcine low density lipoproteins. AB - The order----disorder thermal transition in porcine low density lipoprotein-1 (LDL1) and low density lipoprotein-2 (LDL2) has been studied by differential scanning calorimetry (DSC), 1H-NMR and ESR spin probing. The results by different methods are in general agreement in that the thermal transition in LDL1 takes place at about 32 degrees C. For LDL2 there are two overlapping transition regions, one centered at about 25.5 degrees C and the other at about 33 degrees C. The nature of the two transitions in LDL2 has been discussed. PMID- 2988805 TI - Comparative study of the lipid dynamics in the surface layer of porcine and human high density lipoprotein subclasses by spin labeling. AB - In order to obtain information on the determinants of the lipid dynamics in the high density lipoproteins (HDL), we have compared the structural properties of human HDL subclasses with porcine HDL artificially subdivided into density subfractions corresponding to those of human HDL. Four different positional isomers of spin labeled fatty acids and spin labeled androstanol experienced more restricted motion in porcine HDL than in the human HDL2 and HDL3 subclasses. The differences in the spin label motion could not be accounted for on the basis of the differences in the chemical composition of the lipoproteins examined. They are, however, most probably due to the specific properties of the interactions between lipids and proteins that differ among the lipoproteins. PMID- 2988806 TI - Inhalation studies on the effects of tremolite and brucite dust in rats. AB - Samples of commercially used asbestos, especially chrysotile, are frequently contaminated by small amounts of other fibrous minerals. Among these are tremolite and brucite although pure tremolite is also produced commercially in relatively small quantities. In order to determine how harmful commercially exploited tremolite might be in comparison with other asbestos types and to explore the possibility that small amounts of tremolite and brucite as contaminants could significantly affect the pathogenicity of industrially used chrysotile, long-term animal inhalation and injection studies using rats were undertaken with what were considered to be mineralogically pure samples of these minerals. Rats treated with tremolite developed very high levels of pulmonary fibrosis as well as 16 carcinomas and two mesotheliomas in a group of 39 animals. Tremolite thus proved to be the most dangerous mineral that we have studied. Animals treated with 'brucite' developed moderate levels of pulmonary fibrosis and two carcinomas. Both tremolite and brucite produced mesotheliomas in greater than 90% of animals following i.p. injection. However, it was found that the supposedly pure brucite in fact contained 10% chrysotile, a level of contamination that could well have been responsible for the pathological changes found in both inhalation and intraperitoneal injection studies. The greatest care should be exercised by industry in handling tremolite or materials contaminated with it. PMID- 2988808 TI - Cellular recovery of dividing and confluent C3H10T1/2 cells from N-methyl-N' nitro-N-nitrosoguanidine in the presence of ADP-ribosylation inhibitors. AB - The relationship between treatment with 3-methoxy-benzamide (MBA), a potent inhibitor of ADP-ribosylation reactions, and the response of C3H10T1/2 cells to N methyl-N'-nitro-N-nitrosoguanidine (MNNG) has been examined. Quiescent cells effected potentially lethal damage repair (PLDR) over a 48-h period following MNNG and the repair was coincident with the removal of DNA strand breaks. MBA had no effect on PLDR but was very co-cytotoxic with MNNG in dividing cells. The presence of MBA caused the appearance of an additional number of DNA strand breaks following MNNG in both quiescent and dividing cells. These results suggest that ADP-ribosylation is required for normal cell cycle progression following DNA damage in dividing cells. PMID- 2988809 TI - NAD and the synthesis of (ADP-ribose)n in a human cell strain (46BR) hypersensitive to the lethal effects of 3-aminobenzamide. AB - The cell strain 46BR, derived from an immunodeficient individual, is hypersensitive to the lethal effects of DNA-damaging agents, and of 3 aminobenzamide (3AB), the latter being an inhibitor of the enzyme ADP ribosyltransferase (ADPRT). This hypersensitivity is not found with the noninhibitory analogue, 3-aminobenzoate. The NAD content of 46BR cells is similar to that of fibroblasts from normal human donors, as is the decrease in NAD content following treatment with dimethylsulphate. Both the activity of ADP ribosyltransferase and its inhibition by 3AB in permeabilized cells are similar in 46BR and in normal cell strains. High concentrations of 3AB interfere with purine metabolism in cultured cells. Again this effect is similar in 46BR and normal cells. Thus there is no apparent anomaly either in the activity of ADPRT or in the gross effects of 3AB in 46BR. The sensitivity to 3AB may be caused by a defect in a specific acceptor for the ADP-ribose synthesized by ADPRT, or in some as yet undiscovered action of the inhibitor. PMID- 2988807 TI - Cell cycle perturbations following DNA damage in the presence of ADP-ribosylation inhibitors. AB - Cell cycle analysis by DNA flow cytofluorimetry and autoradiography has been utilized to investigate the effects of 3-methoxybenzamide (MBA), a potent inhibitor of ADP-ribosylation reactions, on cell cycle progression in N-methyl-N' nitro-N-nitrosoguanidine (MNNG)-treated C3H10T1/2 cells. Following a dose of 6.8 microM MNNG, the presence of MBA resulted in an increased length of S phase from approximately 6.5 h to 10 h and in an accumulation of cells in G2 with a mitosis delay of 12 h. Progression to the next S phase occurred 5-10 times more slowly and the cells ultimately accumulated in G2. Increasing the dose of MNNG resulted in a complete block in cell division in the absence of ADP-ribosylation. These results suggest that ADP-ribosylation reactions, which do not seem to be necessary for DNA excision repair in nondividing cells, are essential for coordinating the events of DNA excision repair with DNA replication and events related to progression through the cell cycle. PMID- 2988810 TI - Iodide accumulation into thyroid Golgi vesicles. AB - Pig thyroid Golgi vesicles incubated in a suitable medium were able to concentrate iodide from the medium. This trapping required the integrity of the vesicles, was time- and temperature-dependent, and was inhibited by a competitive inhibitor of iodide active transport (perchlorate), suggesting a facilitated transport mechanism. PMID- 2988812 TI - Phospholipids in plant and animal chromatin. AB - Isolated hepatic nuclei and hepatic chromatin have been analysed for their DNA, RNA, protein and phospholipid content. The protein/DNA ratio is 3 for nuclei and 1.95 for chromatin extracted from Triton X-100 treated nuclei. The phospholipids, (2.36 +/- 0.91 (S.D.) per cent of the total nuclear material), are lost during the chromatin preparation mainly during the Triton X-100 washings of the nuclei. Nevertheless, 10 per cent of the total nuclear phospholipids remain bound to the chromatin. The comparative analysis of both nuclei and chromatin shows a difference in phospholipids and fatty acid composition. Thus, the chromatin associated phospholipid cannot be attributed simply to contaminating nuclear membrane. This is supported by the autoradiographic study of semi-thin sections of interphase nuclei from root apices of Vicia faba in which [3H] ethanolamine is clearly localized in the chromatin and nucleolar regions of the nuclei. PMID- 2988811 TI - Stimulation by cholera toxin of ADP-ribosylation of membrane proteins, adenylate cyclase and insulin release in pancreatic islets. AB - In rat pancreatic islet membranes exposed to [alpha-32P]NAD, cholera toxin stimulated the labelling of three peptides with Mr close to 22 000, 42 000 and 48 000, respectively. In the islets, the toxin-stimulated ADP-ribosylation of the heavy form of the Ns alpha-subunit predominated over that of the light form, in mirror image of the situation found in the exocrine pancreas. When intact islets were preincubated with cholera toxin, the adenylate cyclase activity of a subcellular particulate fraction was increased. The responsiveness of adenylate cyclase to GTP was also augmented, but that to NaF was decreased. In intact islets, the production of cyclic AMP and the glucose-stimulated release of insulin were also enhanced after pretreatment with cholera toxin. These findings reveal the presence in pancreatic islets of the guanyl nucleotide regulatory protein of adenylate cyclase, with an unusual predominance of the heavy form of the Ns alpha-subunit. PMID- 2988813 TI - The plasma membrane sodium-hydrogen exchanger and its role in physiological and pathophysiological processes. AB - The plasma membranes of most if not all vertebrate cells contain a transport system that mediates the transmembrane exchange of sodium for hydrogen. The kinetic properties of this transport system include a 1:1 stoichiometry, affinity for lithium and ammonium ion in addition to sodium and hydrogen, the ability to function in multiple 1:1 exchange modes involving these four cations, sensitivity to inhibition by amiloride and its analogues, and allosteric regulation by intracellular protons. The plasma membrane sodium-hydrogen exchanger plays a physiological role in the regulation of intracellular pH, the control of cell growth and proliferation, stimulus-response coupling in white cells and platelets, the metabolic response to hormones such as insulin and glucocorticoids, the regulation of cell volume, and the transepithelial absorption and secretion of sodium, hydrogen, bicarbonate and chloride ions, and organic anions. Preliminary evidence raises the possibility that the sodium hydrogen exchanger may play a pathophysiological role in such diverse conditions as renal acid-base disorders, essential hypertension, cancer, and tissue or organ hypertrophy. Thus, future research on cellular acid-base homeostasis in general, and on plasma membrane sodium-hydrogen exchange in particular, will enhance our understanding of a great variety of physiological and pathophysiological processes. PMID- 2988814 TI - Stimulation of hypertrophy of cultured neonatal rat heart cells through an alpha 1-adrenergic receptor and induction of beating through an alpha 1- and beta 1 adrenergic receptor interaction. Evidence for independent regulation of growth and beating. AB - Catecholamines may be one of the molecular signals linking increased circulatory demand to myocardial hypertrophy, and I have found previously that norepinephrine stimulates hypertrophy of cultured neonatal rat heart muscle cells through an alpha 1-adrenergic receptor. Since catecholamine stimulation of contractility is believed to be under beta-adrenergic control, I asked whether these cultured heart cells had dual pathways regulating growth and contractility through alpha- and beta-adrenergic receptors, respectively. I examined the effect of adrenergic agents on hypertrophy and beating of myocytes in serum-free cultures. Hypertrophy was defined as an increase in myocyte surface area and in cell protein content, measured by a radioisotopic method, and chronotropic activity was examined visually. Norepinephrine and epinephrine were equipotent stimulants of hypertrophy and beating, increasing cell protein and area 1.5- to 2-fold, and the proportion of beating cells from 5% or less to 95%. Response maxima occurred 24 48 hours after exposure, and EC50 were 20-200 nM. Studies with other agonists (phenylephrine, methoxamine, clonidine, isoproterenol, dopamine) and antagonists (prazosin, terazosin, yohimbine, propranolol, betaxolol, ICI 118,551) indicated that hypertrophy was mediated through an alpha 1-adrenergic receptor, whereas the induction of beating required both alpha 1- and beta 1-receptor activation. Hypertrophied cells with minimal beating were produced by alpha-stimulation, alone. In contrast, alpha-plus beta-stimulation in the presence of cycloheximide to inhibit protein synthesis resulted in maximum beating but no hypertrophy. These findings imply that growth and beating can be regulated independently through separate cellular pathways. PMID- 2988815 TI - The independent effects of oxygen radical scavengers on canine infarct size. Reduction by superoxide dismutase but not catalase. AB - Previous studies demonstrated a significant reduction of ultimate infarct size in the canine heart by the combined administration of superoxide dismutase plus catalase. This study was performed to assess the independent effects of each enzyme on ultimate infarct size due to ischemia/reperfusion. Dogs received 2-hour infusions of superoxide dismutase, catalase, or albumin (controls) via the left atrium beginning 15 minutes before and ending 15 minutes after a 90-minute occlusion of the left circumflex coronary artery. The dogs were killed 6 hours after reperfusion. After histochemical staining, infarct and risk area masses were calculated by gravimetric and planimetric analysis. Infarct size expressed as a percentage of the area at risk was: superoxide dismutase, 19 +/- 5; catalase, 30 +/- 5; and controls, 40 +/- 3. Infarct size in the superoxide dismutase group, but not the catalase group, was significantly less than in controls (P less than 0.05). No significant differences in hemodynamics or area at risk were observed that could explain the differences in infarct size. The results indicate that superoxide dismutase alone protects reperfused ischemic myocardium as well as does the combination of superoxide dismutase and catalase. The beneficial effect of superoxide dismutase and insignificant effect of catalase suggest that tissue damage during ischemia and reperfusion may be mediated largely by superoxide anion but not by hydrogen peroxide. PMID- 2988816 TI - Mechanisms of neurogenic pulmonary edema. PMID- 2988817 TI - Inhibition of aldosterone biosynthesis by atriopeptins in rat adrenal cells. AB - The effect of synthetic atriopeptins on basal and stimulated aldosterone secretion was determined in isolated adrenal glomerulosa cells of the rat. Neither atriopeptin I (1-21) or III (1-24, i.e., the Phe-Arg-Tyr carboxy-terminal extension of atriopeptin I) altered basal aldosterone release. However, if the cells were prepared from adrenals of sodium-depleted rats, the basal aldosterone release was increased by 9-fold, compared with cells from normal rats. This elevated release was inhibited by 32% by atriopeptin I and atriopeptin III. Atriopeptin III was more potent than atriopeptin I. Angiotensin II and adrenocorticotropin stimulated the release of aldosterone in a concentration related manner. Both atriopeptin I and atriopeptin III inhibited the stimulation by the peptides. Atriopeptin I inhibited angiotensin II- and adrenocorticotropin induced aldosterone production by 50% at concentrations of 12 and 11 nM, respectively, and 0.5 and 0.2 nM, respectively, for atriopeptin III. Potassium stimulated aldosterone production was also inhibited by atriopeptin I and atriopeptin III with 50% inhibition at concentrations of 10 and 0.4 nM, respectively. Shorter peptides (1-20, 1-19, and 3-19) were equipotent to atriopeptin I (1-21) as inhibitors of angiotensin II-induced steroidogenesis. To determine the site at which atriopeptins inhibit aldosterone synthesis, we used cyanoketone to inhibit 3 beta-hydroxy-dehydrogenase and dissociate the early and late pathways. Angiotensin II (2 nM) increased the synthesis of pregnenolone (early pathway), as well as the conversion of [3H]corticosterone to [3H]aldosterone (late pathway). Atriopeptin III inhibited basal pregnenolone synthesis by 36% and completely blocked angiotensin II-stimulated synthesis. The peptide similarly inhibited the late pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988818 TI - Nafazatrom-induced salvage of ischemic myocardium in anesthetized dogs is mediated through inhibition of neutrophil function. AB - The effects of nafazatrom on leukocyte function in vitro and in vivo were related to its ability to salvage ischemic myocardium in an occlusion-reperfusion model of myocardial injury in the anesthetized dog. Nafazatrom (0.4-75 microM) produced dose-related inhibition in vitro of neutrophil aggregation, superoxide anion generation, arachidonic acid metabolism, and, to a lesser extent, the release of beta-glucuronidase. In contrast, nafazatrom (0.4-37.5 microM) did not substantially influence platelet aggregation or the platelet metabolism of arachidonic acid. In vivo nafazatrom (10 mg/kg, po) reduced infarct size from 58 +/- 3% of the risk area (mean +/- SEM, n = 9) in control dogs to 23 +/- 2% of the risk area (n = 9, P less than 0.01). Nafazatrom also reduced the incidence of accompanying arrhythmias. Nafazatrom-induced myocardial salvage was not associated with any hemodynamic changes; moreover, it was independent of platelets, since thrombocytopenia did not prevent nafazatrom from exerting a protective effect. Measurements of the neutrophil-specific myeloperoxidase enzyme in ischemic myocardium indicate that the smaller infarct size in dogs treated with nafazatrom is accompanied by diminished leukocyte infiltration. Thus, the ability of nafazatrom to inhibit neutrophil function in vitro and cell infiltration in vivo may underly its myocardial-protective effects. PMID- 2988819 TI - Appearance of superoxide anion radical in cerebral extracellular space during increased prostaglandin synthesis in cats. AB - When increased prostaglandin synthesis was induced in anesthetized cats equipped with cranial windows by topical application of arachidonate (200 micrograms/ml) or bradykinin (20 micrograms/ml), there was reduction of nitroblue tetrazolium, resulting in deposition of the reduced insoluble form of this dye on the brain surface. The amount of reduced nitroblue tetrazolium deposited on the brain surface was measured spectrophotometrically after fixation of the brain by perfusion with aldehydes to eliminate interference from hemoglobin. Topical application of 56 U/ml superoxide dismutase or 20 micrograms/ml indomethacin inhibited nitroblue tetrazolium reduction by 76.5%-82.5% and by 78%-85.5%, respectively. These results show that most of the nitroblue tetrazolium reduction was accounted for by superoxide anion radical generated in the course of arachidonate metabolism via the cyclooxygenase pathway. No superoxide production could be detected in the absence of arachidonate or bradykinin. Histological examination showed no evidence of parenchymal cellular damage or vascular damage and no accumulation of leukocytes. Pronounced leukocyte accumulation occurred 24 hours after topical arachidonate in rabbits with chronically implanted cranial windows. Superoxide appearance was reduced severely by 4,4'-diisothiocyano-2,2' stilbene disulfonate and phenylglyoxal, two specific inhibitors of the anion channel. The most likely explanation for these findings is that increased metabolism of exogenous or endogenous arachidonate via cyclooxygenase results in the appearance of superoxide anion radical in cerebral extracellular space. Superoxide crosses the membrane of undamaged cells via the anion channel. PMID- 2988820 TI - Mechanisms of supersensitivity to sympathomimetic amines in the chronically denervated heart of the conscious dog. AB - Mechanisms of denervation supersensitivity to sympathomimetic amines were studied in conscious animals. Norepinephrine, 0.1 micrograms/kg, increased left ventricular dP/dt significantly more (4208 +/- 828 mm Hg/sec) in dogs with cardiac denervation than in intact dogs (1029 +/- 280 mm Hg/sec), P less than 0.01, whereas responses to isoproterenol were similar in both groups. Denervation supersensitivity to isoproterenol could be demonstrated only after opposing reflex effects were blocked. After ganglionic blockade, norepinephrine still induced 2- to 3-fold greater increases in left ventricular dP/dt and 3- to 7-fold greater increases in heart rate in cardiac-denervated dogs, whereas isoproterenol and prenalterol, not taken up by presynaptic nerve endings, elicited only 40%-50% greater increases in left ventricular dP/dt and heart rate in dogs with cardiac denervation. The density of beta-adrenergic receptors [( 3H]dihydroalprenolol) was elevated (P less than 0.01) in denervated left ventricles (105 +/- 6.9 fmol/mg protein, n = 8) compared to normal left ventricles (70 +/- 6.3 fmol/mg protein, n = 12). This was accompanied by enhanced isoproterenol-mediated adenylate cyclase activity. However, muscarinic cholinergic receptor density, [( 3H]quinuclidinyl benzilate), decreased from control levels of 251 +/- 11 fmol/mg protein (n = 7) to 193 +/- 14 fmol/mg protein (n = 6). Thus, chronic cardiac denervation results in upregulation of the beta-adrenergic receptor and down regulation of the muscarinic receptor. The increased beta-adrenergic receptor density and adenylate cyclase stimulation correlated well with the amount of denervation supersensitivity to isoproterenol and prenalterol, but accounted for only a minor fraction of denervation supersensitivity to norepinephrine. The major mechanism of denervation supersensitivity to norepinephrine appears to involve lack of the norepinephrine reuptake. PMID- 2988821 TI - Quantification of myocardial infarction: a comparison of single photon-emission computed tomography with pyrophosphate to serial plasma MB-creatine kinase measurements. AB - Single photon-emission computed tomography (SPECT) with 99mTc-pyrophosphate (PPi) has been shown to estimate size of myocardial infarction accurately in animals. We tested the hypothesis that SPECT with 99mTc-PPi and blood pool subtraction can provide prompt and accurate estimates of size of myocardial infarction in patients. SPECT estimates are potentially available early after the onset of infarction and should correlate with estimates of infarct size calculated from serial measurements of plasma MB-creatine kinase (CK) activity. Thirty-three patients with acute myocardial infarction and 16 control patients without acute myocardial infarction were studied. Eleven of the patients had transmural anterior myocardial infarction, 16 had transmural inferior myocardial infarction, and six had nontransmural myocardial infarction. SPECT was performed with a commercially available rotating gamma camera. Identical projection images of the distribution of 99mTc-PPi and the ungated cardiac blood pool were acquired sequentially over 180 degrees. Reconstructed sections were color coded and superimposed for purposes of localization of infarct. Areas of increased PPi uptake within myocardial infarcts were thresholded at 65% of peak activity. The blood pool was thresholded at 50% and subtracted to determine the endocardial border for the left ventricle. Myocardial infarcts ranged in size from 1 to 126 gram equivalents (geq) MB-CK. The correlation of MB-CK estimates of size of infarct with size determined by SPECT (both in geq) was good (r = .89 with a regression line of y = 13.1 + 1.5x).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988822 TI - Cyclic blood flow variations induced by platelet-activating factor in stenosed canine coronary arteries despite inhibition of thromboxane synthetase, serotonin receptors, and alpha-adrenergic receptors. AB - The phospholipid platelet-activating factor (PAF) stimulates platelet aggregation and coronary vasoconstriction. In this study we determined whether PAF alters coronary flow patterns in vivo in a canine preparation with concentric coronary artery stenosis. This preparation is characterized by cyclic flow variations in coronary blood flow associated with transient platelet aggregation at the site of the coronary constriction. Thirty-nine male mongrel dogs were used in three protocols. In protocol 1, PAF (10(-9) or 10(-8) mol/min) was infused into the coronary artery proximal to the stenosis to determine (1) whether PAF induces cyclic flow variations and (2) whether PAF has an effect on systemic hemodynamics. Cyclic flow variations were induced in three of six dogs; in these animals, mean arterial pressure decreased by 5.5% and 42.1% 10 min after infusion of the lower and higher dose of PAF. In protocol 2, cyclic flow variations were abolished with either the thromboxane synthetase inhibitor UK38485 (mean dose 2.2 mg/kg iv), the serotonin antagonist ketanserin (0.5 mg/kg iv), or the alpha 2 adrenergic antagonist yohimbine (2 mg/kg iv). Subsequent administration of PAF restored the frequency of cyclic flow variations to the preantagonist levels. Thromboxane (Tx) B2 and 6-keto-PGF1 alpha, the stable metabolites of TxA2 and prostacyclin, respectively, were measured in blood obtained distal to the coronary stenosis. TxB2 levels increased substantially during cyclic flow variations and were returned to control values with the thromboxane synthetase inhibitor UK38485. Infusion of PAF subsequently restored cyclic flow variations without altering coronary arterial coronary arterial TxB2 levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2988823 TI - Urinary adenylate kinase activity as a predictor of renal allograft crises. AB - We examined data on adenylate kinase (EC 2.7.4.3) activity and other clinical chemical values from patients with renal transplants by analysis of variance and discriminant analysis, using various combinations of variables in an attempt to find a predictor of transplant rejection. Some typical data are presented. We conclude that the combination of urinary adenylate kinase and creatinine clearance is the best predictor for identifying patients with transient or destructive renal transplant crises. PMID- 2988824 TI - Use of biotinylated probes for detecting sickle cell anemia. AB - Earlier, we reported that the 5' end of the normal beta-globin gene (beta) resides on a 1.14-kilobase DNA fragment, whereas the 5' end of the sickle cell gene (beta s) resides on a 1.34-kilobase fragment. In that blot hybridization analysis, we used genomic DNA digested with restricted endonuclease Mst II, and radioactive probes with short half-life. We demonstrate here that, if a biotinylated probe is used instead in a slightly modified procedure, sickle cell anemia can be quickly and directly detected if there is as much as 5 micrograms of total genomic DNA in the sample. This procedure obviates the special precautions necessary when radioactive materials are used. PMID- 2988825 TI - More on enzymic assay of inorganic phosphorus. PMID- 2988826 TI - Detection of chemiluminescence in a neutral pH environment. PMID- 2988827 TI - Blood pressure responses to ACTH and adrenaline infusions in dogs. AB - The hypothesis that interactions between adrenaline and adrenal cortical hormones may increase arterial blood pressure has been examined in trained dogs with chronically indwelling aortic and venous catheters. The dogs received continuous infusions for 4 days of ACTH (400 micrograms/day), or adrenaline (8 mg/day) or both ACTH and adrenaline, or saline vehicle. Compared to pre-infusion control values, ACTH raised mean arterial pressure by 14.9 +/- 2.2 mmHg by the 4th day of infusion, adrenaline infused alone had no effect (-0.2 +/- 3.9 mmHg), and ACTH and adrenaline infused together raised pressure 13.1 +/- 3.2 mmHg (not significantly different to ACTH alone). ACTH increased water consumption, Na+ and K+ excretion and creatinine clearance by the 4th day of infusion and these effects were not significantly altered when adrenaline was also infused. Thus adrenaline did not potentiate the hypertension produced by ACTH administration. PMID- 2988828 TI - Myocardial (Na+,K+)-ATPase activity in Dahl salt-sensitive and resistant rats. AB - Vascular (Na+,K+)-pump activity (ouabain-sensitive 86Rb+ uptake) and myocardial (Na+,K+)-ATPase activity are reduced in animals with various forms of low renin, experimental hypertension. On the other hand, vascular (Na+,K+)-pump activity is increased in Dahl salt-sensitive relative to resistant rats (a genetic model of hypertension), regardless of salt intake or blood pressure and it is also increased in Dahl salt-sensitive rats on high salt (8% NaCl) relative to low salt (0.4% NaCl) diets. It has been suggested that this increase in vascular (Na+,K+) pump activity may be secondary to an increase in the vascular sarcolemmal permeability to Na+ in these salt-sensitive rats. In the present study, (Na+,K+) ATPase activity of left ventricular microsomal fractions, was increased in Dahl salt-sensitive relative to resistant rats on low salt diets; however, this difference disappeared when these salt-sensitive and resistant rats were placed on high salt diets. In contrast, myocardial (Na+,K+)-ATPase activity was decreased in Dahl salt-sensitive rats on high relative to low salt diets. Evidence that this decrease in (Na+,K+)-ATPase activity is not secondary to myocardial hypertrophy in the hypertensive salt-sensitive rats, and mechanisms by which decreased cardiovascular (Na+,K+)-pump activity, increased sarcolemmal permeability or both, might contribute to elevated blood pressure, are discussed. PMID- 2988829 TI - Complex glycerol kinase deficiency syndrome explained as X-chromosomal deletion. PMID- 2988830 TI - Immunoglobulin G subclass antibody responses in influenza A and parainfluenza type 1 virus infections. AB - Antibody responses in immunoglobulin G1, G2, G3, G4, A (IgA1) and M isotypes were studied in 10 patients with an acute influenza A and in another 10 patients with a parainfluenza type 1 virus infection using radioimmunoassay with standardized monoclonal anti-immunoglobulins. A four-fold or greater increase of antibody in patients have an acute influenza A virus infection, were found in IgG1 (all 10 cases), IgG3 (seven cases), IgG4 (eight cases) and in IgA1 (six cases) whereas IgG2 and IgM responses were observed only in one and three cases, respectively. The antibody titre values were converted to immunoglobulin units by multiplying the titre by a pre-determined correction coefficient compensating for the varying affinity of the individual monoclonal anti-immunoglobulins. These units were then used to calculate the actual proportions of each isotype. In the convalescent phase, 78% of total anti-influenza A antibodies were estimated to be of IgG1 isotype and other immunoglobulin isotypes varied from 3 to 7% of total. Similar results in parainfluenza virus antibodies were obtained with serum pairs from patients with an acute parainfluenza virus infection. PMID- 2988832 TI - Immune functions in homosexual men with antibodies to HTLV-III in Finland. AB - The occurrence of HTLV-III antibodies in a voluntary group of 175 homosexual men in a low risk AIDS area was studied, and the findings were correlated to clinical, virological, immunological and lifestyle parameters. Fifteen of 175 men had HTLV-III antibodies; two of these had AIDS, five had LAS and two had enlarged lymph nodes. In the HTLV-III antibody negative group, no signs of AIDS or pre AIDS were seen during a 10 month follow-up. In HTLV-III antibody positive individuals, low TH/TS ratio was mainly due to decreased number of TH cells. Most HTLV-III antibody positive cases had low responses to a specific antigen, PPD, while responses to the mitogens PHA and PWM were only slightly affected. In HTLV III antibody negative cases, 13% had a low TH/TS ratio, mostly due to elevation of TS cells. In this group, mitogen and antigen responses were normal or only slightly affected. The results reinforce the causal relationship between HTLV-III and AIDS and suggest that the cells primarily affected by the virus infection are TH cells, responsible for antigen specific responses. Longitudinal studies are required to find out, what is the relationship of immune response to the development of clinical AIDS in HTLV-III infected individuals. PMID- 2988831 TI - Alterations in immunological reactivity in encephalomyocarditis virus-induced murine diabetes. I. Defective primary IgM plaque forming cell responses to sheep erythrocytes: correction by islet cell transplantation. AB - Increasing data suggest a possible viral aetiology of juvenile onset, insulin dependent diabetes mellitus. The M variant of the encephalomyocarditis (EMC) virus infects murine pancreatic beta cells and causes a diabetes like syndrome in susceptible strains of mice. Abnormalities in immunological function have been documented in patients with diabetes mellitus and in spontaneous, streptozotocin induced and alloxan-induced diabetes in animals. The present study documents a significant impairment of the ability of mice with EMC virus (M variant)-induced diabetes to generate a direct, IgM PFC response after in vivo immunization with sheep erythrocytes. This abnormality appears to be a direct consequence of the diabetic state and not EMC virus infection, per se, since mice infected with EMC virus that do not become diabetic have normal direct PFC responses and islet cell transplantation, which cures the diabetes, corrects the defect in PFC responsiveness. PMID- 2988833 TI - Hydrogen peroxide and superoxide production by peripheral blood monocytes in leprosy. AB - Susceptibility to infection with Mycobacterium leprae, the causative organism of leprosy, is the result of a defect in cell-mediated immunity (CMI). The co operation of macrophages and T lymphocytes is known to be essential for competent CMI response. In this study we have examined peripheral blood monocytes from a range of leprosy patients in an attempt to identify a possible defect in macrophage function. The ability of these cells to produce hydrogen peroxide and superoxide, two bactericidal metabolites of the monocyte/macrophage, has been measured. Monocytes from leprosy patients were found to be capable of producing normal amounts of hydrogen peroxide and superoxide, and no differences in production were found between tuberculoid, lepromatous and control monocytes. These results suggest that macrophages in leprosy are competent, and that probably a T lymphocyte defect contributes to susceptibility to this disease. PMID- 2988834 TI - Lysis of varicella zoster virus infected cells by lymphocytes from normal humans and immunosuppressed pediatric leukaemic patients. AB - Varicella zoster virus (VZV) is an important cause of morbidity and mortality in immunosuppressed children but little is known of the cellular mechanisms of VZV immunity. We therefore developed a clinically applicable system to study responses to VZV infected cells. Fresh blood mononuclear cells (MNC) from VZV immune donors killed VZV infected fibroblasts in an 18 h 51Cr release assay. The specificity for virus was confirmed by cold target inhibition. An enhancing role for HLA matching was demonstrated using targets mismatched for HLA, and blocking by antibodies to HLA framework and T cell subsets. Cytotoxicity was not blocked with anti-Ia or anti-VZV antibodies. Killing of VZV infected target cells was reduced in seven out of nine VZV antibody positive patients in remission who were receiving maintenance treatment for acute lymphocytic leukaemia. Three of these patients had normal lymphocyte proliferative responses to VZV. Of the two patients with normal cytotoxic responses to VZV, one had reduced proliferation. It therefore appears that presence of VZV antibody, T cell proliferative responses, and cytotoxicity are independently variable. Cytotoxicity may be more susceptible to immunosuppression than either antibody or T cell proliferation. PMID- 2988835 TI - Further studies on the effects of adrenal steroids in the active transport of serotonin into rat platelets. AB - Male rats were treated with a fixed dose of aminoglutethimide (50 mg/kg s.c.) or with progressively increasing doses (50-100 mg/kg s.c.) for 3 days. Corticosterone levels were found to be decreased in the latter group. Platelet uptake of serotonin as well as the apparent Vmax were decreased, whereas the Km of uptake were increased when compared with that of the control group. Addition of ACTH (10 iu/dl) to control rat platelet and corticosterone (10-80 micrograms/dl) or triamcinolone (0.5-5.0 micrograms/dl) to adrenalectomized rat platelet suspension in vitro did not increase the serotonin uptake of the preparation. Administration of exogenous dexamethasone (0.05-0.2 mg/kg i.m.) or triamcinolone (0.05-5.0 mg/kg i.m.) to adrenalectomized rats, caused a dose related increase in active uptake of serotonin by the platelets. Deoxycorticosterone (0.1-1.0 mg/kg i.m.) did not have this effect. The time course of response to, and the maximum percentage increase in platelet serotonin uptake by, exogenous corticosteroids are related to their glucocorticoid potency. The possible role of glucocorticoids on platelet serotonin uptake process is discussed. PMID- 2988836 TI - Cholinergic neuromodulation by ATP, adenosine and its N6-substituted analogues in guinea-pig ileum. AB - The effects of ATP, adenosine and N6-substituted adenosines, adenosine receptor agonists, on the twitch contraction of guinea-pig ileum evoked by transmural stimulation were evaluated. Adenosine and ATP produced an immediate and concentration dependent inhibition of the twitch, IC50 being 1.1 X 10(-5) mol/l and 1.2 X 10(-5) mol/l, respectively. N6-l-phenylisopropyl adenosine (L-PIA), N6 cyclohexyl adenosine (CHA) and N6-allyl adenosine also induced inhibitions which were gradual and persistent, IC50 being 2.6 X 10(-8), 2.7 X 10(-8) and 5.4 X 10( 7) mol/l, respectively. Dipyridamole (10(-7) mol/l), an adenosine uptake inhibitor, markedly augmented the inhibition evoked by adenosine and ATP, but not that by three N6-substituted adenosines, while theophylline (10(-4) mol/l) almost completely antagonized the inhibitory effects of all purine compounds. IC50 value of adenosine in the presence of dipyridamole (5 X 10(-7) mol/l) was shifted to the left about 50 times from the control, whereas that of L-PIA was virtually unchanged. Tissue-medium ratios indicating uptake of [3H]adenosine, [3H]ATP and [3H]CHA into the segment were 3.23 (s.e.m. = 0.59), 3.59 (s.e.m. = 0.78) and 0.41 (s.e.m. = 0.04), respectively. These results suggest that not only adenosine and ATP but also these N6-substituted adenosines are potent agonists for the P1 receptor, implying a similarity between P1 and A1 receptor in a functional role and these purine compounds may thereby modulate cholinergic neurotransmission by altering adenylate cyclase activity. PMID- 2988837 TI - Correlation of plasma angiotensin II concentration and plasma renin activity during acute hypoxia in dogs. AB - The effects of isocapnic hypoxia on plasma renin activity (PRA) and angiotensin II (AII) concentration were studied in anaesthetized, artificially ventilated dogs. Regression analysis of plasma AII concentration vs PRA, both measured by radioimmunoassay (RIA), was used as an index of converting enzyme activity in vivo. PaO2 decreased from 82 to 30 mmHg but regression analysis did not reveal any inhibition of AII production within the limits of detection of this method (less than 20% inhibition). We conclude that systemic converting enzyme activity, assessed by in vivo measurement and correlation of PRA and AII, is not inhibited by severe hypoxia. PMID- 2988838 TI - Ketanserin does not prevent ACTH-induced hypertension in sheep. AB - The role of serotonin (5HT) in the pathogenesis of ACTH-induced hypertension in sheep has been examined. The pressor responses to injections of 5HT (0.1-30 micrograms/kg) were similar in normotensive and hypertensive sheep. Prior treatment with the 5HT2 receptor antagonist ketanserin had no effect on the development of hypertension produced by ACTH administration. PMID- 2988839 TI - Angiotensin converting enzyme inhibition does not prevent development of ACTH induced hypertension in sheep. AB - The role of the renin-angiotensin system in the onset of ACTH-induced hypertension was examined in five conscious sheep. Captopril infusion alone (15 mg/kg per day) for 2 days produced a small fall in blood pressure. After 2 days of captopril ACTH was infused (20 micrograms/kg per day) for 3 days together with captopril. The blood pressure and electrolyte effects of ACTH administration were not modified by captopril pretreatment. These experiments establish that angiotensin II is not important in the onset of ACTH-induced hypertension in sheep. PMID- 2988840 TI - Role of Ia antigens and interleukin 1 in T-cell proliferation to phytohemagglutinin. AB - Highly purified human T cells were obtained by a four-step purification procedure which included: removal of plastic adherent cells, rosetting with sheep red cells, passage over nylon-wool columns, and treatment with mouse monoclonal antibodies to human Ia antigens and complement. The resulting T cells did not proliferate to phytohemagglutinin (PHA). Purified human interleukin 1 (IL-1) could not substitute for accessory cells in supporting a PHA response. Reconstitution with as little as 0.03% adherent cells resulted in a proliferative response to PHA. T-Cell proliferation to PHA was supported by monocytes, by Ia+ Epstein-Barr virus-transformed lymphoblastoid B-cells lines, and by Ia- cultured human dermal fibroblasts but not by Ia-containing liposomes. Addition of anti-Ia antibodies to monocyte-containing cultures did not inhibit the T-cell proliferative response to PHA. These results suggest that Ia antigen expression by accessory cells is neither necessary nor sufficient to support T-cell proliferation to PHA and that IL-1 is not sufficient to support the proliferation of T cells to PHA. PMID- 2988841 TI - Analysis of antibody response in patients receiving recombinant Escherichia coli derived human interferon-gamma. AB - Recombinant Escherichia coli-derived human interferon-gamma (rIFN-gamma) was given to a total of 20 patients by iv bolus injection at various doses once a week for 4 weeks. The sera obtained 7-10 days after the final injection were analyzed for antibodies against both rIFN-gamma and natural human IFN-gamma. Biological assays demonstrated that the postinoculation sera of the patients did not neutralize the antiviral activities of either rIFN-gamma or natural human IFN gamma. Enzyme-linked immunosorbent assay showed that no detectable antibodies against rIFN-gamma were elicited. These results indicate that this rIFN-gamma preparation is not a potent antigen and may be suitable for longer-term clinical trials and applications in the future. PMID- 2988842 TI - Variables affecting T-lymphocyte subsets in a volunteer blood donor population. AB - Inversion of the normal ratio of helper/inducer (Th) to suppressor/cytotoxic (Ts) T lymphocytes is a characteristic finding in symptomatic and presymptomatic patients with acquired immunodeficiency syndrome (AIDS). As an interim measure to prevent transmission of AIDS via blood transfusion, the Stanford Blood Center performed T-lymphocyte subset analysis as a screening procedure. In this report we present results from 8715 consecutive volunteer blood donors, aged 17-77. The mean Th:Ts ratio was 2.00 +/- 0.70 with a significant trend for increasing Th:Ts with increasing age. In all age groups, donors with antibody to cytomegalovirus (CMV) had lower Th:Ts ratios than CMV-seronegative donors. 1.9% of the donors had Th:Ts values less than or equal to 0.85 and blood from these donors was not used for clinical purposes. Compared to the overall donor population, individuals with Th:Ts less than or equal to 0.85 tended to be male, (79 vs 53%) and CMV sero positive (71 vs 36%); a majority (83%) had a low absolute number of Th cells as well as a low Th:Ts value. Follow-up of donors with low Th:Ts values revealed that some belonged to AIDS high-risk populations, despite denials at the time of donation. One such donor was diagnosed with disseminated Kaposi's sarcoma 8 months after a markedly abnormal T-lymphocyte subset profile was noted during screening. These results suggest that T-lymphocyte subset analysis is capable of identifying at least some blood donors at risk for transmitting AIDS and also point to variable affecting T-cell subsets in apparently healthy blood donors. PMID- 2988843 TI - Fulminant pulmonary coccidioidomycosis in association with Coxsackie B4 infection. PMID- 2988844 TI - Gastrointestinal malacoplakia in children. AB - Four children, whose ages ranged from 1 to 13 years, with malacoplakia of the gastrointestinal tract were treated at King Faisal Specialist Hospital between 1979 and 1983. All patients had either a preceding or a coexisting chronic illness. In one patient, malacoplakia was an incidental finding, while the remaining three patients presented with bloody diarrhea, abdominal pain, recurrent fever, and severe malnutrition. Colonoscopy in two patients revealed markedly inflamed and friable mucosa with focal ulceration alternating with patches of normal mucosa and pseudopolyposis. They were treated with antibiotics and cholinergic agonists. Three patients responded favorably, while one patient continued to have extensive active disease. Although the response to therapy is unpredictable, patients may respond if the treatment is continued on a long-term basis. PMID- 2988845 TI - Treatment of recurrent herpes simplex labialis with topical butylated hydroxytoluene. AB - Butylated hydroxytoluene (BHT) is a hydrophobic compound with in vitro activity against many enveloped viruses, including herpes simplex virus. The effect of topical therapy with 15% BHT in mineral oil on the course of recurrent herpes simplex labialis was examined in 30 patients in a double-blind, placebo controlled pilot study in which treatment was initiated by the physician. Sixteen patients received BHT and 14 received the placebo mineral oil vehicle. The time from lesion onset to dry crust formation was slightly shorter among BHT recipients than among placebo recipients (2.0 and 2.4 days; P = 0.01). Duration of the vesicle-ulcer stages was likewise shorter (1.2 and 2.0 days; P = 0.09), and lesion virus excretion appeared to be less in the subjects who received BHT than in the controls, but these differences were not significant. There was no clinical or laboratory evidence of toxicity. PMID- 2988846 TI - Studies on the mechanism of action of halogenated aromatic hydrocarbons. AB - The halogenated aromatic hydrocarbons (dibenzo-p-dioxins, dibenzofurans, azo[xy]benzenes and biphenyls), a group of toxic chemicals in the environment, (a) are approximate isostereomers; (b) produce a similar pattern of biologic responses and (c) appear to act by a common mechanism. These compounds reversibly bind to a soluble receptor protein to initiate a coordinate gene expression, analogous to the action of steroid hormones. This receptor controls two distinct and dissociable pleiotropic responses: (a) the induction of microsomal monooxygenase activity and other drug metabolizing enzymes and (b) morphologic (i.e. toxic) changes, many of which involve altered cell proliferation and/or differentiation in epithelial tissues. PMID- 2988847 TI - Short course of synacthen therapy as an adjunct in the management of rheumatoid arthritis. AB - The effect of synthetic adrenocorticotrophic hormone (Synacthen), in conjunction with hydroxychloroquine, aurothioglucose, or penicillamine, was evaluated retrospectively in 21 patients with rheumatoid arthritis (RA). One mg of depo Synacthen was administered at increasing intervals of 4 to 14 days for a total period of 3 to 7 months. Fourteen patients with RA on either hydroxychloroquine or aurothioglucose and not on Synacthen, served as controls. Patients in the Synacthen group were, on the whole, sicker, as indicated by a lower functional capacity, higher mean erythrocyte sedimentation rate, and systemic and articular indices. Physicians' estimate of the patients condition after 1 - 2 months of therapy showed no improvement or deterioration in 10 out of 13 cases in the control group. Likewise, the erythrocyte sedimentation rate decreased significantly more and seronegativity was achieved in more of the Synacthen treated cases. Six to 8 months after the beginning of therapy (1 to 4 months after cessation of Synacthen) clinical improvement was comparable in both groups, although seroconversion was more common in patients who had received Synacthen (7 out of 10 as compared to 1 out of 7 respectively). It is suggested that Synacthen may be used safely in the early phase of selected RA patients, until the effect of second-line drugs is achieved. PMID- 2988849 TI - Mechanism of the analgesic effect of calcitonin evidence for a twofold effect: morphine-like and cortisone-like. PMID- 2988848 TI - A multi-systemic disease (lupus-like) preceding bronchioloalveolar carcinoma. AB - A patient with a multi-systemic disease (lupus-like) that preceded the onset of a bronchioloalveolar carcinoma is described, and a brief review of the literature is presented. We suggest that this tumor be listed among the neoplasms responsible for multi-systemic diseases with lupus-like characteristics. PMID- 2988850 TI - Neutrophil mean peroxidase: a biologically interesting parameter with a potential for WBC differential quality control. AB - This study has shown the neutrophil mean peroxidase measurement on the Technicon H6000 to be a simple reproducible technique for measuring neutrophil myeloperoxidase. This measurement shows no interference from contaminating eosinophils unlike spectrophotometric methods. The WBC count had little effect on the CV of the neutrophil mean peroxidase within the normal working range of the machine. Variations in the hydrogen peroxide concentration of the peroxidase substrate also had little effect on this measurement. Haemoglobin however had a significant inhibitory effect on the neutrophil mean peroxidase but this was in a predictable manner. The same inhibition of peroxidase also caused a decrease in the eosinophil count. This varied between individuals. The method described represents a suitable assay for the detection of varying degrees of neutrophil myeloperoxidase activity including the acquired and hereditary deficiencies. This parameter is potentially useful for the quality control of the cytochemically derived WBC differential. PMID- 2988851 TI - Malignant pleural effusions. AB - Various diseases of the gastrointestinal tract at times are accompanied by an exudative pleural effusion. The exudative pleural effusions resulting from esophageal perforation, pancreatic disease, subphrenic abscess, intrahepatic abscess, splenic abscess, abdominal operations, and diaphragmatic hernia are discussed in this article. PMID- 2988852 TI - Collagen accumulation in the neonatal rat skin: absence of fibrillar collagen degradation during normal growth. AB - Collagen-bound collagenase activity was assayed in the entire skins of growing neonatal rats. The levels of fibrillar collagen degradation were found to be extremely low throughout the first six days of life. Less than one percent of the collagen accumulated during one day's growth could be degraded by the collagenase bound to the extracellular fibrils of the skin. Control experiments, which included the addition of purified rat uterine collagenase to the skins both before and after homogenization, showed that collagenase activity is easily detectable in the tissue when it is present. It therefore appears that the vast majority of skin collagen, once deposited as fibrils in the skin, fails to turn over during growth. Support for this concept was provided by experiments in which neonatal animals were injected with the potent synthetic glucocorticoid, triamcinolone. Very low levels of collagen-bound collagenase, comparable to those observed in control animals, were found in the steroid-treated animals. Furthermore, the inhibition of collagen synthesis in these animals resulted in a constant chemical content of collagen as well as a constant amount of proteinaceous [14C]-hydroxyproline after injection of [14C]-proline over a 72 hour period. Our results strongly suggest that the bulk of fibrillar collagen does not participate in a dynamic equilibrium between synthesis and degradation during normal neonatal growth. In addition, the results in steroid-treated animals suggest that the rate of collagen accumulation during this period appears to be essentially a function only of collagen synthesis. PMID- 2988853 TI - Human skin fibroblast collagenase: interaction with substrate and inhibitor. AB - Human skin fibroblasts secrete collagen, procollagenase and a collagenase inhibitor. This study addresses the nature of the interaction between these important fibroblast products. The binding of procollagenase and of active collagenase to native collagen in solution was examined by employing Sephadex G 150 gel-filtration chromatography to separate bound versus unbound enzyme. Active enzyme bound readily to collagen; the equilibrium constant of binding, Kd, was calculated to be 5.1 to 10(-7)M. Thus, collagenase binds with nearly equal affinity to both monomeric collagen and aggregated fibrils (Kd = 9 X 10(-7)M; [Welgus et al., 1980]). Furthermore, since Kd congruent to Km congruent to 10( 6)M, the ratio k2/k1 must be extremely small, directly implicating the catalytic step represented by the rate constant k2, and not the binding of enzyme to substrate, as the rate-limiting step of collagenase action. In contrast, procollagenase demonstrated no capacity to bind to collagen. The interaction of procollagenase and of active collagenase with inhibitor was examined utilizing both conventional and high-precision liquid gel-filtration chromatography. A higher molecular weight complex could be demonstrated consisting of active collagenase and inhibitor; no such interaction occurred between procollagenase and the inhibitory protein. Analysis of Lineweaver-Burk plots showed that inhibition was accompanied by a corresponding change in Vmax; Km remained unchanged. Such results are indicative of a noncompetitive mechanism of inhibition and are consistent with the formation of an enzyme-inhibitor complex. The Ki of enzyme-inhibitor binding was determined to be less than 10(-9)M. The data indicate that procollagenase can neither interact with its specific inhibitor nor bind to collagen. Extracellular activation of the collagenase zymogen is thus a critical event, which can be followed either by binding to substrate or interaction with inhibitor. PMID- 2988854 TI - Mononuclear cell-conditioned medium containing mononuclear cell factor (MCF), homologous with interleukin 1, stimulates collagen and fibronectin synthesis by adherent rheumatoid synovial cells: effects of prostaglandin E2 and indomethacin. AB - Adherent rheumatoid synovial cells produce and release into supernatant culture medium latent collagenase and PGE2. The levels of collagenase and PGE2 can be increased by a soluble factor present in mononuclear cell-conditioned medium, partially purified by gel-filtration, which has homologies with interleukin 1, and is produced by monocyte/macrophages. The synovial cell cultures produce collagens (procollagens) and fibronectin as well. The factor(s) present in the mononuclear cell conditioned medium which increases medium levels of collagenase PGE2 also stimulates synthesis of total protein as well as types I and III procollagen by the synovial cells. This stimulation by the monocyte factor is augmented in the presence of indomethacin, which blocks endogenous PGE2 production. Medium levels of fibronectin parallel those of procollagen. The addition of exogenous PGE2 abolishes the effect of indomethacin on collagen and fibronectin synthesis. These observations of mononuclear cell-mediated increases in fibronectin synthesis may account for the high levels of fibronectin found by others in rheumatoid synovium and synovial fluids as the increases in collagen synthesis might also explain the fibrosis observed in some rheumatoid joints. PMID- 2988855 TI - Metabolism of glycerate-2,3-P2--VII. Enzymes involved in the metabolism of glycerate-2,3-P2 in cat tissues. AB - The levels of the enzymes involved in the metabolism of glycerate-2,3-P2 (phosphoglycerate mutase, bisphosphoglycerate synthase-phosphatase and bisphosphoglycerate phosphatase) in cat and in pig tissues are different. The main difference is the low level of bisphosphoglycerate synthase-phosphatase in cat tissues. As a consequence, in contrast with pig erythrocytes, in cat erythrocytes, both the synthesis and the breakdown of glycerate-2,3-P2 are mainly controlled by phosphoglycerate mutase. PMID- 2988856 TI - Phytase activity in chicken erythrocytes and its control by organic phosphates (glycerate-2,3-P2 and inositol-P5) during avian development. AB - The activity and basic kinetic constants of phytase were studied in chicken erythrocytes during animal development. The regulatory inhibition of phytase by IHP and 2,3-BPG takes place at key stages of the development. As in mammals, there is a specific control of the levels of organic phosphate involved in the oxygenation process of haemoglobin, during animal development. PMID- 2988857 TI - Chemical modification of the active site of the NADP-linked glutamate dehydrogenase from Trypanosoma cruzi. AB - The NADP-linked glutamate dehydrogenase (NADP-gluDH) purified from epimastigotes of the Tulahuen strain, Tul 2 stock, of Trypanosoma cruzi, was inhibited by Cibacron Blue FG3A, and inactivated by preincubation with phenylglyoxal or Woodward's Reagent K. The inhibition by Cibracron Blue FG3A, competitive towards NADPH with an apparent Ki of 20 microM, suggests that the enzyme presents the "dinucleotide fold" characteristic of most dehydrogenases and kinases. The inactivation of the NADP-gluDH by preincubation with phenylglyoxal, with a reaction order of 1, and the partial protection afforded by alpha-oxoglutarate, suggest the presence of one arginine residue in the active site of the enzyme, which might participate in the binding of alpha-oxoglutarate through interaction with one of the carboxyl groups of the substrate. The inactivation of the NADP gluDH by preincubation with Woodward's Reagent K suggests the presence of a carboxyl group, from an aspartic or glutamic acid residue, at the active site, which might participate in the binding of the cationic substrate NH+4. The presence of NADPH during preincubation with the reagent increased the inactivation rate, which suggests that binding of the coenzyme increases the exposure of the reactive carboxyl group. PMID- 2988858 TI - Hormone-sensitive lipase from swine adipose tissue: identification and some properties. AB - Swine adipose tissue hormone-sensitive lipase, purified 475-fold to 10% protein purity, has been identified as a polypeptide of Mr = 84,000. The enzyme has high specific activity against tri-, di- and monoacylglycerols, as well as cholesterol esters, and is inhibited by millimolar NaF, and micromolar HgCl2 and DFP. The enzyme polypeptide serves as a substrate for cyclic AMP-dependent protein kinase. The characteristics of the hormone-sensitive lipase from swine adipose tissue are similar to those reported previously for the enzyme from rat. They differ from those reported for the lipase from chicken adipose tissue, and possible reasons for these differences are discussed. PMID- 2988859 TI - The use of a logistic model for the quantitative interpretation of indirect sandwich enzyme labelled immunosorbent assays (ELISA) for antibodies and antigens in foot and mouth disease. AB - A three parameter logistic model is described for the analysis of profiles of optical density vs log dose for indirect sandwich ELISA tests in foot and mouth disease. The model describes the observed phenomenon of saturability with increasing dose, and its parameters can be interpreted in terms of molecular binding events. A computer program to fit the model is described. An approximate statistical test is developed which can be used to test for departures from equivalence for replicate profiles. It is found that correlation of the optical density values to a standard reference reaction considerably improves reproducibility. PMID- 2988860 TI - Developmental cystic renal neoplasms in children. Diagnostic imaging characteristics. AB - We report diagnostic imaging and histopathologic findings in four children with different cystic renal neoplasms. The features discerned with computed tomography (CT), as well as ultrasound, correlated well with gross histologic findings, although a definitive diagnosis could not be made on the basis of CT and ultrasound alone. Because the most malignant variants of these lesions cannot be excluded preoperatively, we advocate complete diagnostic imaging studies, followed by nephrectomy, for all cases of cystic renal neoplasms in infants and children. PMID- 2988861 TI - Human metallothionein-II processed gene is located in region p11----q21 of chromosome 4. AB - Metallothionein (MT) genes comprise a multigene family encoding low-molecular weight, heavy-metal-binding proteins. We have mapped a human MT-II processed gene to chromosome 4, using Southern blotting in combination with a human X mouse hybrid clone panel containing defined subsets of human chromosomes. We have further localized this gene to region p11----q21, using in situ hybridization. PMID- 2988862 TI - Chromosomal site of hepatitis B virus (HBV) integration in a human hepatocellular carcinoma-derived cell line. AB - The single site of integration of hepatitis B virus in the human hepatocellular carcinoma cell line Hep 3B 2-1/7 was found to segregate with human chromosome 12 in somatic cell hybrids. Analysis of metaphase spreads of Hep 3B 2-1/7 following in situ hybridization with pHBV revealed integration at 12q13----q14, a location that coincides with a fragile site, fra (12q13). The possible significance of this location to the development of hepatocellular carcinomas is discussed. PMID- 2988863 TI - Transbronchial needle aspiration in the diagnosis of submucosal and peribronchial bronchogenic carcinoma. AB - Although exophytic endobronchial lesions can readily be diagnosed by routine forceps biopsy through the fiberoptic bronchoscope, submucosal or peribronchial tumor can be difficult to diagnose with nonsurgical techniques. We evaluated the utility of transbronchial needle aspiration (TBNA) through the fiberoptic bronchoscope in 31 patients presenting with endoscopic abnormalities suggestive of submucosal or peribronchial tumor. TBNA was performed using a 20 g X 1 cm needle, followed by forceps biopsy of the same area. Forceps biopsy was positive in 17 cases (55 percent) and TBNA in 22 (71 percent) (p = .302). The combination of forceps biopsy and TBNA was positive in 27 cases (89 percent), which was significantly better than forceps biopsy alone (p = .00195). In addition, the wash or the brush detected three additional carcinomas, so the combination of TBNA, forceps biopsy, wash, and brush had a diagnostic yield of 97 percent. We conclude that TBNA significantly increases the yield over forceps biopsy alone in the detection of submucosal or peribronchial bronchogenic carcinoma and that the maximal diagnostic yield is obtained by the combination of TBNA, forceps biopsy, wash, and brush when appropriate endoscopic abnormalities are encountered. PMID- 2988865 TI - Male breast cancer. PMID- 2988864 TI - Nasopharyngeal mucosal changes in EB virus VCA-IgA antibody positive persons. PMID- 2988866 TI - Primary carcinoma of the trachea. Report of six cases. PMID- 2988867 TI - Oropharyngeal shedding of infectious Epstein-Barr virus in healthy virus-immune donors. A prospective study. PMID- 2988868 TI - Primary low malignancy tracheobronchial tumor. PMID- 2988869 TI - Effect of postoperative radiotherapy on the results of radical mastectomy for stage II breast cancer. PMID- 2988870 TI - CSF and PB lymphocyte subpopulations in sporadic encephalitis. PMID- 2988871 TI - Cytomegalovirus and Epstein-Barr virus infection in renal transplant recipients. PMID- 2988872 TI - [Factors related to the therapeutic responses of ovarian malignancies]. PMID- 2988873 TI - [Immunologic function of ovarian cancer]. PMID- 2988874 TI - [Pathological analysis of 1726 cases of ovarian tumors]. PMID- 2988875 TI - [Serum lactic dehydrogenase assay in the diagnosis and therapeutic monitored of ovarian carcinoma]. PMID- 2988877 TI - Centromeric satellite DNA in the newt Triturus cristatus karelinii and related species: its distribution and transcription on lampbrush chromosomes. AB - Two abundant satellite DNA sequences have been identified in and cloned from the DNA of Triturus cristatus karelinii. The smaller of these with a repeat unit of 33 base pairs (bp) is designated TkS1, the larger with 68 bp is designated TkS2. These satellites are also present in DNA from T.c. cristatus, T.c. carnifex and T. marmoratus but in substantially lower copy number. In situ hybridisations to lampbrush chromosomes of T.c. karelinii and T.c. cristatus have shown that the satellites are concentrated in the heterochromatic centromere bars of T.c. karelinii and in a region around the centromere granule in T.c. cristatus. The satellites also bind specifically to the centromere regions of mitotic metaphase chromosomes. They do not bind to the heteromorphic arms of chromosome 1, which have previously been shown to be rich in highly repeated DNA. DNA/RNA-transcript in situ hybrids to lampbrush chromosomes with TkS1 suggest that this sequence is occasionally transcribed on lampbrush loops near the centromeres. PMID- 2988876 TI - Gene amplification in Djungarian hamster cell lines possessing decreased plasma membrane permeability for colchicine and some other drugs. AB - By multistep selection a set of clones and sublines possessing different levels of resistance to colchicine or adriablastin was obtained from the SV40 transformed Djungarian hamster cell lines, DM-15 and DMcap. Resistance to both colchicine and adriablastin is associated with an alteration of plasma membrane permeability leading to a decreased uptake of various drugs (3H-colchicine, 3H cytochalasin B, 3H-actinomycin D, 3H-puromycin, 3H-vinblastine, 14C chloramphenicol). The DNA of cells highly resistant to cholchicine can transmit resistance only to low dosages of the drug. Comparison of DNAs from wild-type and resistant cells digested by restriction endonucleases revealed new classes of repeated DNA sequences in resistant cell lines. The degree of DNA repetition was correlated with the level of drug resistance. The repeated DNA sequences evidently represent parts of the genome that are amplified in resistant cells. The size of the amplified sequences is 200-250 kilobase pairs (kb). Cell lines highly resistant to colchicine contain amplified DNA, which like mitochondrial DNA replicate asynchronously with the main portion of the cellular DNA and related but not identical DNA sequences are amplified in independent cell lines selected for resistance to colchicine, adriablastin, and actinomycin D. These cell lines display similar patterns of alterations of plasma membrane permeability. The amplified DNA sequences may contain a gene or genes the overexpression of which leads to change in plasma membrane permeability and a development of resistance to various drugs. PMID- 2988879 TI - STb enterotoxin of Escherichia coli: cyclic nucleotide-independent secretion. AB - Escherichia coli may produce a heat-labile enterotoxin (LT) or two heat-stable enterotoxins (STa, STb). Experimentally, STb is consistently active only in 5 h weaned pig intestinal loops (WPIL), an effect that is largely removable by rinsing. At least three mechanisms initiate small intestinal secretion: cyclic AMP (LT), cyclic GMP (STa) and calcium (A23187). All three increase short-circuit current (SCC) in Ussing chambers by stimulating net Cl- secretion. STb significantly increases SCC within 2-5 minutes in Ussing chambers and is independent of cyclic AMP and cyclic GMP. When compared to crude culture filtrates of a non-toxigenic strain of E. coli, crude culture filtrates of STb did not alter Na+ or Cl- undirectional or net fluxes. However, the calculated residual ion flux (JRnet) increased significantly in STb-treated tissues and appeared to largely account for the STb-induced increase in SCC. Furosemide applied serosally (10(-3) M), the removal of extracellular calcium, and lanthanum chloride (10(-3) M) did not inhibit the effect of STb on SCC. Chlorpromazine (0.4 mM) completely inhibited STb-induced secretion in porcine loops. This inhibition was a non-specific reversal of the STb effect because in Ussing chambers, chlorpromazine simply induced an equal and opposite effect on SCC. These results indicate that STb initiates intestinal secretion in porcine jejunum in vitro by stimulating primarily non-chloride anion secretion in the absence of extracellular calcium. We postulate that STb causes bicarbonate secretion by a mechanism distinct from those of previously studied enterotoxins. PMID- 2988880 TI - [Pulmonary arterial pressure and shunt in pigs in experimental hypoxia. II. Determination of blood histamine, cAMP and cGMP in hypoxia-induced pulmonary hypertension]. PMID- 2988878 TI - Coding and potential regulatory sequences of a cluster of chorion genes in Drosophila melanogaster. AB - We have characterized at the nucleotide level a 4.8-kilobase pair segment of the third chromosome of Drosophila melanogaster, which contains a cluster of three chorion genes, s18-1, s15-1 and s19-1. These genes are tandemly oriented and share the same basic organization: a small and a large exon separated by a short intron in the signal peptide region. In the coding region, limited similarities at the DNA and protein level suggest a common but distant evolutionary origin. The flanking sequences were searched for elements that might be involved in controlling the tissue-specific and temporally regulated expression and the selective amplification of the chorion genes. A good candidate for a cis regulatory element is the hexamer, TCACGT, which is found in all three genes in a highly significant position, 23 to 27 nucleotides upstream of the TATA-box, accompanied by additional, less exact similarities. Palindromes and short inverted repeats that are found in the vicinity of their complement are non uniformly distributed: they are most concentrated in the 3' flanking part of all three genes, in and near regions of unusually high A and T content. The highest number of dyad symmetries, reminiscent of sequences that function as viral replication origins, is found associated with the T- and A-rich regions between genes s18-1 and s15-1. PMID- 2988881 TI - Factors influencing local recurrence after abdominoperineal resection for cancer of the rectum. AB - Pelvic and perineal recurrences of cancer after rectal amputation are frequent, often isolated, and thus directly responsible for a fatal outcome by local decompression accidents or infection. This study explores the patterns of recurrence after "curative" operation for rectal cancers. One hundred thirteen patients who underwent abdominoperineal resection are reviewed: there were 36 local recurrences, i.e., an incidence of 31.8 percent. About 70 percent of these recurrences occurred within two years after surgical treatment. Low level of tumor in the rectum, local spread into perirectal fat or serosa, lymph node involvement, and histologic grade of malignancy were the only factors that were statistically related to local recurrence. The strategy for careful follow-up of patients at risk is outlined and a plea is made for a controlled trial of postoperative radiotherapy. PMID- 2988882 TI - Small-cell carcinoma of the rectum. Clinicopathologic study. AB - Three cases of small-cell carcinoma of the rectum are presented. Our conclusions are as follows: small-cell carcinoma and carcinoid tumor of the rectum might have a common origin; indications for surgical intervention for small-cell carcinoma of the rectum should be considered seriously; small-cell carcinoma of the rectum should be classified as an aggressive type of rectal tumor that metastasizes easily to distant organs. PMID- 2988883 TI - Stem cell carcinoma of the colon and rectum. Report of two cases and review of the literature. AB - Two cases of highly malignant tumors, one originating in the sigmoid colon and the other in the rectum, are presented. Both tumors showed light microscopic, electron microscopic, and immunohistochemical evidence of multidirectional differentiation. The tumors were composed mainly of undifferentiated cells, but focally merging into areas with adenocarcinomatous and squamous differentiation. Ultrastructurally and histochemically, a predominant endocrine differentiation was present in the undifferentiated areas of the tumors. These two cases lend further support to the recent concept of a pluripotential stem cell in the mucosa of the gastrointestinal tract capable of differentiation in several directions. PMID- 2988884 TI - Villous adenoma depletion syndrome. Evidence for a cyclic nucleotide-mediated diarrhea. AB - Massive secretory diarrhea is associated with some villous adenomas. The mechanism of this secretion is unknown but the character of the diarrhea resembles that of cyclic nucleotide-mediated diarrheas. We have compared the cyclic nucleotide metabolism of a large secretory villous adenoma with a nonsecretory villous adenoma, a solid carcinoma and their normal mucosae. The adenylate cyclase, cyclic AMP content, and a cyclic AMP-dependent protein kinase ratios in the secretory tumor were increased as compared to these values in the nonsecretory tumors and normal mucosae, a situation similar to that seen with cholera toxin-induced diarrhea. Our data suggest that the massive diarrhea in our patient with a secretory villous adenoma may be related to increased adenylate cyclase activity. PMID- 2988885 TI - Obstructive uropathy. An unusual presentation of primary linitis plastica of the colon. AB - A patient with primary linitis plastica of the colon has been presented. The unusually aggressive nature of this tumor is demonstrated in a young man who presented with obstructive uropathy. Extensive renal involvement is a rare complication of this disease, and obstructive uropathy is a previously undescribed presentation. If early recognition is to favorably alter prognosis, then awareness of this form of colonic malignancy is essential. PMID- 2988886 TI - Small bowel neoplasm. PMID- 2988887 TI - Flattening postprandial blood glucose responses with guar gum: acute effects. AB - It has been proposed that high-carbohydrate, high-fiber (HCF) diets might serve as useful therapeutic modality in non-insulin-dependent diabetes mellitus (NIDDM). One problem in evaluating clinical trials of this therapy is that, by their very nature, the trials cannot be double blinded. We have developed HCF and placebo granola-type bars using complex absorbable carbohydrate and guar gum fiber to circumvent this methodologic problem. The HCF bars, when consumed with an ad lib. diet, assure an HCF intake without imposing other dietary restrictions. To test the short-term efficacy of the bars, 9 normal adult volunteers, 2 women with impaired glucose tolerance, and 20 patients with NIDDM consumed the bars alone or with meals. Blood glucose responses when HCF bars were consumed alone were blunted when compared with the placebo response (P less than 0.0005 to P less than 0.002), with the most marked suppression occurring in the early postprandial period. In contrast, when the bars were consumed along with breakfast, HCF and placebo responses were virtually identical in the early postprandial period, but showed a progressively greater difference from 90 to 240 min (P less than 0.02 to P less than 0.0005). When consumed with lunch as well as breakfast, the HCF bars caused flattening of blood glucose responses during the late postprandial period after breakfast and maintained flattened responses during the early and late postprandial periods after lunch (P less than 0.05 to P less than 0.005). It is concluded that these HCF bars can be used to blunt postprandial blood glucose responses, in subjects with either normal or abnormal carbohydrate metabolism. PMID- 2988888 TI - Glucose-induced alterations in nerve metabolism: current perspective on the pathogenesis of diabetic neuropathy and future directions for research and therapy. AB - Recent animal and in vitro studies have identified several interrelated metabolic abnormalities in diabetic nerve that are attributable to elevated ambient glucose concentrations. In combination, these metabolic changes may induce a variety of biochemical and biophysical alterations in peripheral nerve that are highly relevant to the pathogenesis of diabetic neuropathy. This article reviews the current status of several of these metabolic defects and describes ways in which their interaction could lead to pathogenetically important changes in nerve metabolism, function, and structure. Areas of related future research are also discussed. PMID- 2988889 TI - Isolated ACTH deficiency contributing to frequent hypoglycemia in type I diabetes. PMID- 2988890 TI - Use of artificial endocrine pancreas in postoperative management of patients with beta-cell hyperfunction. PMID- 2988891 TI - Mutational analyses of the Moloney murine sarcoma virus enhancer. AB - Retroviral enhancers seem to play an integral role in determining tissue tropism. In an attempt to define the sequences essential for Moloney murine sarcoma virus (Mo-MSV) enhancer activity, we have undertaken deletion and insertion mutational analyses of the 72-bp repeats. We demonstrate that, in contrast to simian virus 40 (SV40), the proximal 19 bp of the 72-bp unit of Mo-MSV are not essential for enhancer function in mouse fibroblasts. Our analysis further localizes the 5' boundary of the Mo-MSV enhancer to a 5'-CAGGAT-3' region. PMID- 2988892 TI - Identification by exonuclease footprinting of a distal promoter-binding protein from HeLa cell extracts. AB - Incubation of a HeLa cell fraction termed P1000 with genomic DNA fragments containing portions of the 5'-flanking region of the ovalbumin gene, followed by digestion with exonucleases, shows that this cell fraction contains a protein (or proteins) that binds to the distal promoter of the ovalbumin gene. The protein protects both strands and spans a minimum of 22 bp on the noncoding strand from nucleotides -68 to -90. The 5' border of the protected region on the coding strand is located at nucleotide -81. A similar site on the chick beta-globin gene is also protected by HeLa cell fraction P1000. The exonuclease footprinting methods used to identify the distal promoter-binding protein are more sensitive than the DNaseI footprinting method, and can be used to identify sequence specific binding proteins in a mixture of DNA binding proteins. PMID- 2988893 TI - [Pathology of small cell lung carcinoma (SCLC)]. PMID- 2988894 TI - [X-ray manifestations of small cell lung carcinoma (SCLC). An analysis of 345 cases]. PMID- 2988895 TI - [Double infusion chemotherapy by hepatic artery and portal vein in primary malignant tumor of the liver. Report of 18 cases]. PMID- 2988896 TI - [Observation on sister chromatid exchange (SCE) in patients with lung cancer]. PMID- 2988897 TI - [Action of chelators of divalent cations on the receptor potential of the retinal rods of the frog]. PMID- 2988899 TI - [Internal homology in the primary structure of the polyprotein of the poliovirus: the possible existence of 2 viral proteinases]. PMID- 2988898 TI - [Proton magnetic resonance spectra of lymphocytes in experimental leukemia]. PMID- 2988900 TI - [Temperature-sensitive mutants of Drosophila with altered cAMP phosphodiesterase activity: has the gene for calmodulin been found?]. PMID- 2988901 TI - [Effect of irradiation with monochromatic visible light on the cAMP content in mammalian cells]. PMID- 2988902 TI - [Effect of X-ray-induced DNA breaks on the hypersensitivity of the minichromosomes of SV40 virus to DNAse I]. PMID- 2988903 TI - beta-Lactamase inhibitors. AB - Numerous beta-lactamase inhibitors have been developed to overcome the problems of bacterial resistance to beta-lactam antibiotics. One such agent, clavulanic acid, has been combined with amoxicillin, and the combination is effective against many amoxicillin-resistant organisms. Clinical studies have demonstrated the efficacy of amoxicillin/clavulanate in a variety of infections, however, comparative studies are few and have not demonstrated sufficiently the superiority of the combination over conventional antibiotic therapy. In addition, side effects appear to be more frequent than with amoxicillin alone. The place in therapy of amoxicillin/clavulanate is discussed. PMID- 2988904 TI - [Endocrinological reactions following UV A whole body irradiation]. AB - The basal secretion of proteo-hormones alpha-MSH and ACTH in plasma as well as the changes of the plasma concentrations following UV A-whole-body irradiation were investigated on 40 young male volunteers with different pigmentation levels (Caucasians: skin types I, II, III. Blacks: skin type VI). Significantly lower mean basic values of alpha-MSH and ACTH of light-haired persons in comparison with dark-haired and black persons (p less than 0,05) were demonstrated. We observed furthermore a significant increase of these proteo-hormones (alpha-MSH: skin type I: 26,7%, skin type II: 22,7%) in persons less pigmented within a short time after UV A whole-body irradiation in contrast to the more pigmented volunteers. These results prove a cutaneous peripheral sensor for UV A-rays, reacting with a different sensitivity depending on disposition and inducing endocrinological reactions. How this cutaneous-hypothalamic-pituitary stimulus mediation functions in detail is not completely revealed up to now. In what respect the present results, which can be explained as a consequence of evolutionary development, have a connection with the induction of melanoma remains to be seen. PMID- 2988905 TI - [New aspects of the vibration disease. Results of a study in a large factory]. AB - The vibration syndrome includes the frequently observed polysystemic symptom complex, and also encompasses dermatological symptoms as well. Under conditions where vibration and noise exceed the acceptable norms, a very high illness index must be reckoned with. Aside from the characteristic symptoms in the periphery and the vascular and bone systems, numerous symptoms in the central nervous system (still not recognised as occupational disease) have been diagnosed. The present study was carried out by the Cracaw Medical Academy, Poland, who performed medical examinations on the employees of large companies. The results of these examinations are linked to pathogenic vibration. Because of these findings, new recommendations were made concerning changes needed in prophylactic examinations. PMID- 2988906 TI - [Occurrence of enzootic bovine leukosis in Papua New Guinea]. PMID- 2988907 TI - [Human papillomavirus (HPV) and carcinoma of the uterine cervix]. PMID- 2988908 TI - [Physiological reactions to trauma]. PMID- 2988909 TI - [Immunity to embryonal stage-specific antigens in viral carcinogenesis]. AB - The oncogenesis induced by the monkey SA7(C8) adenovirus in CBA/Ca mice has shown that immune responses to embryonal antigens are formed at early stages of the latent period and are preserved for a long time reaching the maximum by the 30th day of the latent period. The observed immune response to early embryonal antigens is considered as a factor of the tumour growth immunostimulation and also as a condition limiting the antitumour immunity. PMID- 2988910 TI - The role of calcium/phospholipid-dependent protein kinase in Leydig cell steroidogenesis. AB - Purified rat Leydig cell cytosol was found to contain a protein kinase which was dependent on the presence of both calcium and phospholipids (phosphatidylserine and diolein), i.e. calcium/phospholipid-dependent protein kinase. The peak of Ca/phospholipid-dependent protein kinase was separated from type I and type II cAMP-dependent protein kinase by DE-52 chromatography. 4 beta-Phorbol-12 myristate-13-acetate (PMA), a tumor-promoting agent, could substitute for diolein in activation of Ca/phospholipid-dependent protein kinase. PMA caused dose dependent increments of testosterone formation by Leydig cells, whereas inactive phorbol esters had no significant effects. PMA-induced testosterone formation was dependent on extracellular calcium and could be blocked by the addition of the calcium channel-blocking agent nifedipine. Since PMA can directly activate Ca/phospholipid-dependent protein kinase and increase testosterone formation, these results suggest that Ca/phospholipid-dependent protein kinase may be involved in modulating Leydig cell steroidogenesis in addition to the classical cAMP-dependent protein kinase pathway. PMID- 2988911 TI - Androgen inhibition of follicle-stimulating hormone-stimulated luteinizing hormone receptor formation in cultured rat granulosa cells. AB - Since LH receptors are decreased in atretic follicles known to contain high androgen levels, we have studied the androgen modulation of LH receptor formation in vitro. Granulosa cells from hypophysectomized, diethylstilbestrol-treated rats were cultured for 3 days with FSH in the presence or absence of nonaromatizable androgens, dihydrotestosterone and 5 alpha-androstane-3 alpha, 17 beta-diol, or a synthetic androgen, R1881 (17 beta-hydroxy-17 alpha-methyl-4,9,11-estratrien-3 one). FSH increased LH receptor content in granulosa cells, while concomitant androgen treatment decreased LH receptor content in a dose- and time-dependent manner, without changing the equilibrium dissociation constant (Kd) for human CG. R1881 (10(-7) M), dihydrotestosterone (10(-6) M), and 5 alpha-androstane-3 alpha, 17 beta-diol (10(-6) M) inhibited LH receptor content by 68%, 65%, and 65%, respectively. Similar to earlier findings, these androgens enhanced FSH stimulated progesterone biosynthesis and aromatase activity in the same cells. To study their LH responsiveness, androgen-treated cells were washed and reincubated for 2 more days with or without LH. Although basal progesterone production was elevated by R1881 pretreatment, the androgen-pretreated cells were less responsive to LH. Treatment with cyanoketone, an inhibitor of 3 beta hydroxysteroid dehydrogenase, did not alter the inhibitory effects of R1881 on LH receptors, indicating that the androgen action is not mediated by endogenous progestins. Furthermore, R1881 inhibited the stimulation of LH receptor formation by forskolin, cholera toxin, and 8-bromo-cAMP, suggesting that androgens may inhibit LH receptor induction by affecting post-cAMP events. Estrogen treatment enhanced the FSH induction of LH receptor content, while concomitant addition of R1881 also suppressed the estrogen action. Thus, androgens inhibit FSH-induced functional LH receptors in cultured rat granulosa cells. The androgen effect is exerted, at least partially, at post-cAMP sites and is independent of changes in progestin biosynthesis. PMID- 2988912 TI - Angiotensin and converting enzyme regulate extrarenal salt excretion in ducks. AB - Numerous previous studies have proposed a salt-conserving role for the renin angiotensin system in mammals, but there is little evidence of this putative role in birds. Especially interesting are marine birds, which have a relatively limited ability to regulate the osmolality and ionic composition of their urine but possess extrarenal salt glands capable of excreting a highly concentrated NaCl solution. In the present experiments, hypertonic saline, angiotensin I (ANG I), and captopril were infused iv into chronically cannulated ducks to study the neuroendocrine, cardiovascular, renal, and extrarenal excretory responses to osmotic stress. Infusion of hypertonic saline elicited nasal salt excretion, which could be stopped completely by coadministration of ANG I. The effective dose of ANG I increased the plasma norepinephrine (NE) concentration, but did not alter heart rate or arterial blood pressure. Captopril enhanced extrarenal salt excretion in the saline-loaded ducks. The converting enzyme inhibitor also blocked the noradrenergic and NaCl-retaining actions of ANG I; conversely, coadministration of captopril and ANG I increased the plasma epinephrine (E) concentration. These findings indicate that the renin-angiotensin system, in addition to effects on the sympathoadrenal system, regulates NaCl and water metabolism in birds with extrarenal salt glands. PMID- 2988914 TI - The effect of Streptococcus pneumoniae infection on the binding of triiodothyronine to nuclei isolated from rat liver. AB - Hepatic nuclei were isolated from control and Streptococcus (S.) pneumoniae infected rats to determine the effect of S. pneumoniae infection on the binding capacity and affinity of hepatic nuclei for T3. Infection did not affect the purity or yield of the isolated nuclei. A significant decrease in serum total T3 was observed 40 h after inoculation with S. pneumoniae. Serum free T3 was significantly decreased by 20 h after inoculation, but returned to control levels by 40 h after inoculation. Scatchard analysis of nuclear T3 binding, determined under optimal conditions, confirmed the presence of high affinity, low capacity sites for T3 on nuclei isolated from both control and infected rats. During infection, the maximum binding capacity of the purified nuclei for T3 decreased significantly, but infection had no significant effect on the affinity of the receptor for T3. These results suggest that the decrease in serum T3 observed during infection is not contributed to by increased hepatic nuclear T3 receptor concentration and that the increased hepatic RNA, protein, and lipid synthesis observed during S. pneumoniae infection in the rat is not the result of increased binding or affinity of T3 to the nuclear receptor. PMID- 2988913 TI - Receptors for epidermal growth factor in the rat uterus. AB - Uterine membranes contain high affinity [dissociation constant (Kd) = 0.36 nM], saturable binding sites for [125I]iodo-epidermal growth factor (EGF). The binding of [125I] iodo-EGF is specific since it is abolished by excess unlabeled EGF but not by excess unlabeled insulin, fibroblast growth factor, or multiplication stimulating activity. Incubation of [125I] iodo-EGF with uterine membranes, followed by chemical cross-linking with disuccinimidyl suberate and detergent extraction reveals a major species of specifically bound EGF (mol wt = 170,000) and a minor species (mol wt = 150,000) visualized by autoradiography of sodium dodecyl sulfate gels after electrophoresis of the extracts. In detergent solubilized preparations EGF also stimulates the phosphorylation of major (mol wt = 170,000) and minor (mol wt = 150,000) species of identical molecular weight. The increased phosphorylation produced by incubation of membrane extracts with EGF occurs largely at tyrosine residues, as indicated by phosphoamino acid analysis. These results indicate that the rat uterus contains high affinity EGF binding sites with the properties expected of EGF receptors. PMID- 2988915 TI - Positive regulation of the guinea pig thyrotropin receptor. AB - Subcutaneous administration of bovine (b) TSH (up to 10 IU) to 8-week-old male guinea pigs was followed by a transient elevation in serum thyroid hormone levels (T4 and T3) and an increase in thyroid weight (approximately 25-40%), which returned to control levels by 3 days. Total thyroid TSH receptor content was assessed by the binding of receptor-purified [125I]bTSH to 15,000 X g fractions of thyroid homogenate. The TSH receptor content paralleled the increase in thyroid weight, with no detectable change in the TSH-binding capacity per mg tissue. Intraperitoneal minipump infusions of bTSH (1 IU/day) over 6 days produced marked and persistent increases in thyroid hormone levels and thyroid weight (greater than 300%) and a similar increase in TSH receptor content. There was no change in the single site equilibrium association constant for bTSH [1.1 X 10(9) M-1 +/- (SE) 9.6 X 10(7) M-1] and no alteration in the binding capacity per mg tissue (67.2 +/- 6.4 pg/mg). Investigation of the in vitro adenylate cyclase response to bTSH (10 mU/ml) and the production of immunoassayable cAMP showed no difference between thyroid tissue obtained from bTSH-treated animals and that obtained from untreated control animals. These observations demonstrated that TSH exerted a positive regulatory effect on its receptor and, under the in vivo conditions used, failed to induce TSH receptor loss or physiologically important desensitization. Such data may explain how TSH receptor antibodies are able to act as TSH agonists and maintain increased thyroid hormone output in human disease. PMID- 2988916 TI - Corticotropin-releasing factor stimulation of adrenocorticotropin and beta endorphin release: effects of acute and chronic stress. AB - The effects of acute and chronic stress on the release of ACTH and beta-endorphin in response to stimulation by ovine corticotropin-releasing factor (CRF) and arginine vasopressin were examined. Pituitaries were removed from rats who had received either acute stress, chronic stress daily for 14 days with the last stress occurring 24 h before decapitation, or chronic stress followed by an acute stress immediately before decapitation (chronic stress-acute stress). Pituitaries from naive unstressed animals were used as the control group. After processing into single cell suspensions, the pituitaries were incubated with various doses of CRF (10(-11) M to 10(-9) M) and AVP (10(-10) M to 10(-8) M). Release of ACTH and beta-endorphin into the medium was measured by RIA. A clear dose-dependent response to both releasers was seen in control pituitaries. In acute stress, a decreased responsiveness to arginine vasopressin and CRF was seen. This same blunted response was not seen in chronic stress even if the animals are stressed immediately before decapitation. At higher doses of CRF (10(-9) M) a substantially increased release of ACTH and beta-endorphin was seen in the chronically stressed rats. When the content of the anterior pituitary lobe was assayed in these animals, both chronic stress groups show increased content of ACTH and beta-endorphin, which may indicate an increase amount of ACTH and beta endorphin in the releasable pools in chronic stress. In addition, the failure of further stress to alter the response to CRF in the chronic stress-acute stress group may indicate a down-regulation of the steroid feedback on the pituitary. However, it is clear that no down-regulation of the CRF receptor occurs in this chronic stress paradigm. PMID- 2988917 TI - Adenosine differentially amplifies luteinizing hormone- over follicle-stimulating hormone-mediated effects in acute cultures of rat granulosa cells. AB - Adenosine has been shown to acutely amplify LH-dependent events in luteal cells and FSH-dependent events in granulosa cells. In this study, the specificity of purines on mature rat granulosa cell ATP levels in short term culture was assessed, and a comparison of the relative effect of adenosine on amplification of FSH- and LH-stimulated cAMP accumulation was made. Adenosine rapidly and significantly increased ATP levels in granulosa cells. This effect was maximal (approximately 2-fold) within 60 min of culture and occurred in the absence or presence of FSH or LH. The increase in granulosa cell ATP levels by adenosine was dose dependent, with half-maximal and maximum responses of 10 and 30 microM adenosine, respectively. Dipyridamole (10 microM), a purine transport inhibitor, blocked the adenosine-dependent increase in granulosa cell ATP levels. Adenosine and 5'-AMP were equipotent in increasing cell ATP levels; adenine also increased ATP levels, but was significantly less active (approximately 50% of adenosine), whereas hypoxanthine, inosine, and xanthine were inactive. FSH was consistently found to decrease granulosa cell ATP levels by about 30% in the absence or presence of adenosine, whereas LH had no effect on cell levels of ATP. Both FSH and LH significantly stimulated cAMP accumulation in granulosa cells, but the maximal response to FSH was substantially greater than that to LH. Adenosine significantly amplified cAMP accumulation in response to both FSH and LH, but the effect of adenosine on this response to FSH was modest. Amplification by adenosine of cAMP accumulation in response to LH was substantial and about 2- to 3-fold greater than that seen with FSH. These studies show that purines acutely and specifically increase ATP levels in rat granulosa cells. Since adenosine augments LH-dependent cAMP accumulation to a greater extent than FSH-stimulated cAMP production, we suggest that adenosine may favor premature follicular luteinization and, perhaps, function as a mediator of atresia in the developing follicle. PMID- 2988918 TI - Cytosol activator protein from rat reticulocytes requires the stimulatory guanine nucleotide-binding protein for its actions on adenylate cyclase. AB - Rat reticulocytes contain a cytosol activator protein (RCAP) that augments catecholamine-sensitive adenylate cyclase activity in reticulocyte membranes. A highly purified preparation of RCAP, obtained by Sephacryl S-200 chromatography, was used to elucidate further its mechanism of action. The specific activity of the S-200 fraction to augment isoproterenol responsiveness was increased approximately 1,100-fold over the starting material, from 1.2 to 1,300 nmol cAMP formed per mg RCAP. The mol wt of RCAP is approximately 20,000. The effect of RCAP to enhance isoproterenol responsiveness was apparent within 20 sec, virtually abolishing the normal lag time of hormone-activated adenylate cyclase. In addition to its effects on catecholamine-responsive adenylate cyclase, RCAP significantly increased basal [21 +/- 3 (+/- SEM) to 41 +/- 4 pmol/mg protein X 30 min; P less than 0.02], guanyl-5'-yl-imidodiphosphate-associated (3173 +/- 213 to 4339 +/- 365 pmol/mg X 30 min; P less than 0.03), and fluoride-associated (5152 +/- 64 to 5807 +/- 58 pmol/mg X 30 min; P less than 0.05) adenylate cyclase activities. RCAP also altered the characteristics of agonist binding to the beta adrenergic receptor of reticulocyte membranes, causing an increase in the apparent IC50 for isoproterenol from 0.7 +/- 0.2 to 7.9 +/- 1.6 microM (P less than 0.001). Similar to its effects on reticulocytes, RCAP enhanced isoproterenol and prostaglandin E2-sensitive adenylate cyclase activity in the wild-type S49 lymphoma cell and shifted the binding isotherm for isoproterenol rightward. In cyc-, the mutant that lacks the stimulatory guanine nucleotide-binding protein (Ns) and in UNC, the mutant in which receptors are uncoupled from N, RCAP was ineffective. Moreover, RCAP decreased agonist affinity for the beta-adrenergic receptor in wild-type S49 cells, but not in cyc- or UNC cells. These observations suggest that RCAP requires a functional Ns unit for its effects on hormone sensitive adenylate cyclase activity. PMID- 2988919 TI - Effects and pharmacokinetic properties of the rat/human corticotropin-releasing factor in rhesus monkeys. AB - The biological effects and pharmacokinetic properties of the recently sequenced rat/human corticotropin-releasing factor (r/hCRF) were evaluated in the rhesus monkey and compared to those of the previously studied ovine corticotropin releasing factor (oCRF). An iv bolus of 0, 0.1, 1, 10, and 100 micrograms/kg r/hCRF and 1 microgram/kg oCRF were given to rhesus monkeys (four to five tests per dose). Serial blood samples were drawn before and up to 180 min after the injection for determination of plasma immunoreactive (IR) ACTH, cortisol, and IR r/hCRF or IR-oCRF concentrations. Mean arterial blood pressure and heart rate were monitored. r/hCRF stimulated ACTH and cortisol secretion in a dose-dependent fashion. Its potency was similar to that of oCRF. r/hCRF, however, had a shorter half-life and a 3-fold higher MCR than oCRF. A dose-dependent decrease in the MCR of r/hCRF was observed, which may indicate a saturation of the clearance mechanisms. Significant decreases in mean arterial blood pressure, increases in heart rate, and a profound facial flush occurred at the dose of 100 micrograms/kg r/hCRF. We conclude that r/hCRF stimulates ACTH and cortisol secretion in a nonhuman primate with a potency similar to that of oCRF. The peptide has marked hypotensive effects at high doses and is cleared 3 times more rapidly than oCRF. PMID- 2988920 TI - Protein kinase stimulation of steroidogenesis in rat luteal cell mitochondria. AB - The regulatory role of cAMP-dependent protein kinase in steroidogenesis was examined in luteal cell mitochondria prepared from heavily luteinized prepubertal rat ovaries. The cAMP-dependent protein kinase, localized in luteal mitochondria, comprised 5.5% of the total cellular protein kinase activity (cAMP-dependent). Intact mitochondria supported by a suitable electron-donor substrate and inhibited by isoxazole converted cholesterol to a single steroid product, pregnenolone. Neither (Bu)2 cAMP nor a crude preparation of cytosolic protein kinase stimulated pregnenolone production from cholesterol when added to intact luteal cell mitochondria; however, mitochondria treated with 10 mM CaCl2 became responsive to both (Bu)2 cAMP and protein kinase by showing increased pregnenolone production. Likewise, the addition of cytosol protein kinase to incubations of cholesterol and crude cholesterol sidechain cleavage enzyme (cytochrome P-450cscc) isolated from luteal mitochondria, also stimulated pregnenolone production. Cholesterol-poor mitochondria, depleted of endogenous sterol by prolonged preincubation, when subsequently incubated with Ca+2 plus (Bu)2 cAMP and protein kinase showed significantly increased pregnenolone production. Conversely, mitochondria with greatly increased intramitochondrial cholesterol after preincubation with 200 microM cholesterol and a cytochrome P 450cscc inhibitor (aminoglutethimide) synthesized pregnenolone in significantly higher amounts than either normal or cholesterol-poor mitochondria after removal of the aminoglutethimide block. However, addition of (Bu)2cAMP or protein kinase to Ca+2-treated cholesterol-rich mitochondria failed to additionally stimulate pregnenolone synthesis. We conclude from these observations that the mitochondrial membrane normally excludes protein kinase and (Bu)2cAMP from any stimulatory action on cholesterol side-chain cleavage. Disruption of the mitochondrial membrane by high Ca+2 concentrations eliminates this barrier and permits (Bu)2cAMP and protein kinase stimulation of the CSCC enzyme system. The mechanism of stimulation is not clear. It could involve direct action on the CSCC enzyme. Alternatively, an increase in either intramitochondrial transport or binding of cholesterol substrate to the CSCC enzyme could be facilitated by protein kinase action. Direct stimulation of the enzyme by protein kinase seems less likely, since increased enzyme activity was not observed in the presence of high concentrations of intramitochondrial cholesterol substrate. PMID- 2988921 TI - Hypophysiotropic regulation of adrenocorticotropin secretion in response to insulin-induced hypoglycemia. AB - The hypophysiotropic coding of ACTH secretion resulting from insulin-induced hypoglycemia was investigated in urethane-anesthetized fasted rats. The participation of corticotropin-releasing factor (CRF), arginine vasopressin (AVP), and catecholamines in the ACTH response was first investigated by systemic administration of CRF antiserum, an AVP pressor antagonist, or a ganglionic blocking agent. These treatments were without effect on the hypoglycemic response, which was characterized by a 67% fall in systemic glucose levels within 30 min of insulin administration. ACTH secretion in response to insulin-induced hypoglycemia was differentially affected by these pharmacological treatments. Administration of antiserum to CRF abolished the ACTH response, whereas ganglionic blockade was without significant effect. However, administration of a vasopressinergic pressor antagonist significantly attenuated ACTH secretion after insulin treatment. These observations suggested the participation of both CRF and AVP in mediation of the ACTH secretory response to hypoglycemia. Infusion of glucose to counter the hypoglycemia action of insulin injection prevented the ACTH secretory response. Measurement of immunoreactive (ir) CRF, irAVP, and ir oxytocin in sequential collections of hypophysial portal plasma revealed a significant elevation of irAVP concentration without concomitant elevation of irCRF or ir-oxytocin levels. We propose that CRF functions in a permissive role, maintaining a relatively constant portal concentration and thereby allowing expression of the weaker ACTH-releasing activity of AVP and other secretagogues. Thus, AVP, not CRF, appears to represent the dynamic mediator of ACTH secretion accompanying insulin-induced hypoglycemia. These observations provide additional support for the hypothesis of multifactor stimulus-specific hypophysiotropic coding of ACTH secretion. PMID- 2988922 TI - Factors affecting the conversion of androstenedione to estrogens by human fetal hepatocytes in monolayer culture. AB - The purpose of the present investigation was to characterize and determine what hormones affect the activity of aromatase in human fetal hepatocytes maintained in primary monolayer culture. The major product of aromatization of androstenedione was estrone sulfate. Optimal conditions for assay of aromatase activity in fetal liver cells were determined. The apparent Km for androstenedione was 50 nM. Aromatase activity was stimulated by glucocorticoids in the presence of fetal calf serum. The concentration of dexamethasone required for half-maximal stimulation was 10(-8) M, similar to the concentration required for half-maximal binding to glucocorticoid receptors. This action of dexamethasone was inhibited by cortisol 21-mesylate, a glucocorticoid antagonist. Aromatase activity was also stimulated by (Bu)2cAMP and cholera toxin, and was inhibited by fetal calf serum. This effect of fetal calf serum was mimicked by epidermal growth factor. However, epidermal growth factor did not mimic the permissive action of serum to stimulate aromatase activity by dexamethasone. In these respects, the regulation of aromatase activity of human fetal hepatocytes is similar to that of human adipose stromal cells. A polycyclic hydrocarbon, benzo(a)pyrene, which causes induction of aryl hydrocarbon hydroxylase activity in fetal hepatocytes, inhibited the stimulation of aromatase activity by dexamethasone. Of a number of hormones tested, including glucagon, insulin, angiotensin II, ACTH, hCG, GH, PRL, and T3, only glucocorticoids were effective in stimulating aromatase activity of human fetal hepatocytes. These results emphasize the complex and multiparameter nature of the regulation of aromatase activity in this as in other tissues. PMID- 2988923 TI - Preliminary characterization of enterooxyntic activity on the guinea pig oxyntic cell. AB - Stimuli originating in the small intestine enhance gastric acid secretion, an effect termed the intestinal phase of gastric secretion. A humoral agent, enterooxyntin (EO) may mediate this effect. Mechanical distension of an ex vivo perfused segment of canine jejunum caused the release of EO measured by cytochemical quantification of hydroxyl ion production (HIP) in guinea pig gastric oxyntic cells, an index of acid secretion. The H2 receptor antagonist, cimetidine (10(-5) M), and diamine oxidase, which hydrolyzes endogenous histamine, reduced HIP by 60% and 48%, respectively. Atropine (10(-5) M), the muscarinic cholinergic antagonist, reduced HIP by 75%. EO-induced HIP was also inhibited partially by the Ca2+ antagonist EGTA (10(-6) M) and by blockade of calcium channels with LaCl3 (10(-6) M). EO does not appear to operate through any single pathway. EO may be a single substance, different from gastrin, or a mixture of substances that have stimulatory effects on the oxyntic cell. Its action on the oxyntic cell is apparently mediated by both histaminergic and cholinergic pathways. Since neither cimetidine nor atropine completely inhibited EO-induced HIP, a direct effect of EO on the oxyntic cell seems likely and appears to depend on Ca2+. Isolation and purification of EO will be necessary to better assess the potency, efficacy, and the detailed cellular mechanisms of EO action. PMID- 2988924 TI - Adrenocorticotropin receptors in rat adrenal glomerulosa cells. AB - The results presented here demonstrate, for the first time, the presence of ACTH receptors in the zona glomerulosa of adrenal glands. We obtained the surprising result that the glomerulosa cells carry a higher concentration of ACTH receptors than the fasciculata cells. The analog [Phe2,Nle4]ACTH was iodinated by the iodogen method and separated by HPLC; it was obtained carrier-free and has a specific activity of 600 muCi/micrograms, retaining full biological potency. After 30 min of incubation at 22 C for concentrations of 2 X 10(-11) M [125I]ACTH, specific binding values were 4.85 +/- 0.44% (n = 15) and 1.85 +/- 0.18% (n = 15), respectively, for 50,000 glomerulosa or fasciculata cells. For the glomerulosa, our results indicated a density of 6.5 X 10(4), receptors/cell of the high affinity type (Kd1 = 7.6 X 10(-11) M) and 1.0 X 10(6) receptors of the low affinity type (Kd2 = 1.2 X 10(-9) M). In the zona fasciculata, we found 7.2 X 10(3) receptors of high affinity (Kd1 = 1.1 X 10(-11) M) per cell and 6.3 X 10(5) of low affinity (Kd2 = 2.9 X 10(-9) M). The dissociation constant for the high affinity site of the glomerulosa cells was in excellent correlation with the half-maximal stimulation dose of ACTH for aldosterone and corticosterone (Kd1 = 7.6 X 10(-11) M vs. ED50 of 8 X 10(-11) and 3 X 10(-11) M). Results from primary cultures showed a decrease in binding capacity after 1 day in culture and then an increase to the initial value after 3 days in culture. PMID- 2988926 TI - Pressor-type vasopressin receptors in the adrenal cortex: properties of binding, effects on phosphoinositide metabolism and aldosterone secretion. AB - Specific, high affinity sites that bound tritium-labeled arginine-vasopressin (3H AVP) were detected in a crude membrane fraction of rat adrenal capsules (chiefly zona glomerulosa). Binding displacement experiments with peptide analogs of AVP suggested that the binding site is a pressor (V1) type receptor for AVP. When added to dispersed rat adrenal glomerulosa cells, vasopressin (10(-8)-10(-6)M) stimulated the incorporation of 32P-phosphate into phosphatidylinositol, and the effect was blocked by the AVP receptor antagonist peptide d(CH2)5Tyr(Me)AVP. Vasopressin also increased the breakdown of phosphatidylinositol-4,5-bisphosphate within 1 min after its addition to the incubation medium. Superfused zona glomerulosa cells responded to AVP (10(-8)-(-6)M) by increasing their aldosterone production. The response could be blocked by the antagonist peptide. These data show that functionally active V1 receptors are present in rat glomerulosa cells, and suggest that vasopressin may regulate the function of the adrenal glomerulosa. PMID- 2988925 TI - Epidermal growth factor receptor levels in mouse mammary glands in various physiological states. AB - Experiments were undertaken to demonstrate and characterize specific receptors for epidermal growth factor (EGF) in mammary glands of female BALB/c mice in various physiological states. The results of an in vitro desaturation technique are also presented which allow estimation of the total EGF-binding sites per mg membrane protein. Binding of the ligand [125I]iodo-EGF is both time and temperature dependent. Maximum binding to the membrane is achieved after 6 h of incubation with [125I]iodo-EGF at 23 C. Scatchard analysis of equilibrium binding using membrane preparations of mammary glands from virgin mice yields two classes of high affinity receptors with Kd values of 0.8 +/- 0.1 and 5.0 +/- 0.4 X 10( 10) M and receptor concentrations of 10 +/- 1.2 and 23.5 +/- 2 fmol/mg protein, respectively. Membrane preparations of mammary tissues from cycling, gestating, and lactating mice were used to correlate cellular receptor levels to the physiological state of the animal. Beginning at weaning, there is a constant decrease in high affinity receptor level with increasing age, as well as through the early stages of both gestation and lactation. On day 10 of gestation, receptor levels increase, reaching 15.2 +/- 1.6 fmol/mg protein, followed by a decrease to 3.8 +/- 0.9 fmol/mg protein on day 10 of lactation. We conclude that membrane preparations from the mouse mammary gland contain specific high affinity receptors for EGF, and that receptor levels are characteristic of the physiological state. PMID- 2988927 TI - Growth hormone-releasing factor binding sites in rat anterior pituitary membrane homogenates: modulation by glucocorticoids. AB - Specific high affinity binding sites for growth hormone releasing factor (GRF) were described in rat anterior pituitary homogenates with use of the analog [His1 mono-125I-Tyr10,Nle27]-hGRF(1-32)-NH2 as radioligand. Computerized analysis of competition experiments indicated one class of specific high affinity binding sites with a dissociation constant of 0.19 nM. The relative binding affinities of rGRF, hGRF(1-40) and various analogs correlated well with their in vitro biological potencies. Further, the number binding sites was drastically decreased after adrenalectomy; chronic dexamethasone treatment of these animals restored GRF binding capacity to control without changing binding affinity. These results may in part explain the enhanced responsiveness of the somatotroph after dexamethasone treatment. PMID- 2988928 TI - Association of ulcerative colitis, sclerosing cholangitis and cholangiocarcinoma in a patient with IgA deficiency. AB - A case of an unusual association of ulcerative colitis, sclerosing cholangitis, cholangiocarcinoma and IgA deficiency in a 22-year-old man is reported. The patient was admitted to the hospital with jaundice. Ulcerative colitis had been diagnosed 4 years before. The association of sclerosing cholangitis and cholangiocarcinoma was demonstrated. In addition IgA deficiency was discovered. Possible pathogenetic implications are discussed. PMID- 2988929 TI - The heart and the atrial natriuretic factor. AB - The search for natriuretic hormones or factors by studies of negative pressure breathing, atrial distension experiments, head-out water immersion, expansion of blood volume, Na+/K+-ATPase inhibitors and parabiosis experiments in Dahl rats has led to the finding that the atria are a peptide-secreting endocrine gland. This new natriuretic hormone has now been purified, sequenced and synthetized, and its cDNA and gene have been cloned. The native and synthetic hormones exert identical wide ranging effects (possibly through particulate guanylate cyclase stimulation and adenylate cyclase inhibition) on the kidney, blood vessels, adrenal cortex, and pituitary. Physiopathologic implications of the hormone in experimental hypertension, congestive heart failure, and expansion of blood volume are beginning to emerge. PMID- 2988930 TI - Immune mechanisms in Graves' disease. PMID- 2988931 TI - Interaction of high density lipoproteins with cholesteryl ester-laden macrophages: biochemical and morphological characterization of cell surface receptor binding, endocytosis and resecretion of high density lipoproteins by macrophages. AB - Morphological and biochemical experiments were carried out to investigate the interaction of human serum high density lipoproteins (HDL) with mouse peritoneal macrophages. It is demonstrated that resident mouse peritoneal macrophages express HDL receptors. Subsequent to receptor-mediated binding, HDL are internalized and intracellularly transported into endosomes. These endosomes do not fuse with the lysosomal compartment but interact with the margin of intracellular plasma lipid droplets. Macrophages do not degrade, but rather resecrete internalized HDL particles as described for the transferrin-receptor pathway. HDL binding to freshly isolated macrophages is saturable at a concentration of approximately 320 ng HDL-protein/mg cell protein and a Scatchard plot indicates the presence of some 130 000-190 000 receptors/cell with a Kd of approximately 9 X 10(-7) M. Binding of HDL on the macrophage surface is significantly enhanced in cholesterol-laden macrophages, whereas the increase in the rate of uptake and secretion is less pronounced. Within the HDL fraction the HDL2 subclass showed higher binding, uptake and secretion activity as compared with HDL3. From these experimental data we postulate that cholesterol uptake from macrophages is mediated by HDL particles which interact with these cells via a receptor-mediated retroendocytosis pathway. PMID- 2988932 TI - The spatial and temporal pattern of beta NGF receptor expression in the developing chick embryo. AB - To gain insight into the developmental program of nerve growth factor (NGF) receptor expression, the binding of [125I] beta NGF to frozen chick sections was investigated autorradiographically between embryonic day 3 (E3) and post-hatching day 3. Strong NGF receptor expression was observed as early as E4, throughout embryonic development and in the post-hatching period at the classical NGF target sites: the paravertebral sensory and sympathetic ganglia, the paraaortal sympathetic ganglia as well as the cranial sensory ganglia with neurons of neural crest origin and their respective nerves. Only weak [125I] beta NGF binding was observed during a restricted time span in the parasympathetic ciliary ganglion. Clear differences were observed in the intensity and in the developmental time course of [125I] beta NGF binding to the dorsomedial and ventrolateral aspects of the dorsal root ganglia. NGF receptors were also found to be expressed on central axons of the dorsal root entry zone and the dorsal tract in the spinal cord. A transient expression of specific NGF binding sites of the same high affinity as measured at the classical NGF targets, was detected in the lateral motor column and in muscle at the time of motoneuron synapse formation and elimination. PMID- 2988933 TI - Defective splicing of thyroglobulin gene transcripts in the congenital goitre of the Afrikander cattle. AB - The structure of thyroglobulin mRNA was analyzed in an inbred herd of Afrikander cattle with hereditary goitre. Northern transfer of RNA from affected animals revealed both a shorter (approximately 7100 bases) and a normal-sized (approximately 8200 bases) thyroglobulin mRNA when hybridized to bovine thyroglobulin cDNA clones. S1 nuclease mapping experiments established that 1100 bases are deleted in the 5' region of the smaller mRNA. Electron microscopy of RNA from animals with goitre hybridized to a bovine genomic DNA clone showed that the region deleted corresponds to exon 9 of the thyroglobulin gene. Southern blot analysis of the exon 9 region revealed differences between affected and control animals with the enzymes PstI and TaqI. Although they could reflect a linkage disequilibrium between the mutation and restriction fragment length polymorphism, it is noteworthy that these differences map in the region of the exon 9/intron 9 junction. Our results show that a genetic lesion in the thyroglobulin gene causes aberrant splicing of the pre-mRNA, and suggest that the responsible mutation is at the exon 9/intron 9 junction. PMID- 2988935 TI - The human ubiquitin multigene family: some genes contain multiple directly repeated ubiquitin coding sequences. AB - Ubiquitin coding sequences were isolated from a human genomic library and two cDNA libraries. One human ubiquitin gene consists of 2055 nucleotides and codes for a polyprotein consisting of 685 amino acid residues. The polyprotein contains nine direct repeats of the ubiquitin amino acid sequence and the last ubiquitin sequence is extended with an additional valyl residue at the C-terminal end. No spacer sequences separate the ubiquitin repeats and the coding regions are not interrupted by intervening sequences. This particular gene is transcribed since cDNAs corresponding to the genomic sequence have been isolated. At least two more types of ubiquitin genes are encoded in the human genome, one coding for an ubiquitin monomer while another presumably codes for three or four direct repeats of the ubiquitin sequence. Human DNA contains many copies of the ubiquitin sequence. Ubiquitin is therefore encoded in the human genome as a multigene family. PMID- 2988934 TI - SB subregion of the human major histocompatibility complex: gene organization, allelic polymorphism and expression in transformed cells. AB - The SB region of the human major histocompatibility complex (MHC) has been cloned from cosmid and lambda phage libraries made from the human B-lymphoblastoid cell line Priess (DR4/4, DC4/4, SB3/4). Two alpha genes and two beta genes are encoded in the 100 kb long SB region in the order SB alpha-SB beta-SX alpha-SX beta. The SB alpha and SB beta genes encode the alpha and beta subunits of the SB subset of class II MHC molecules. Both the SX alpha and the SX beta genes are pseudogenes in the haplotype examined. From the isolated clones, the two haplotypes of the Priess cell line, SB3 and SB4, are distinguished by nucleotide sequencing and blot hybridization analyses. Restriction site polymorphisms between the SB3 and SB4 clones were observed only in relatively small regions of the SB beta and SX beta genes. A mouse macrophage cell line was transfected with one of the cosmid clones containing both SB alpha and SB beta genes. Expression of the alpha and beta genes was detected by fluorescene-activated cell sorting (FACS) and two dimensional gel electrophoresis using SB-specific monoclonal antibodies. PMID- 2988936 TI - Queuosine modification of the wobble base in tRNAHis influences 'in vivo' decoding properties. AB - The 'in vivo' decoding properties of four tRNAHis isoacceptors, two from Drosophila melanogaster and two from brewer's yeast, were studied after their microinjection, along with turnip yellow mosaic virus (TYMV) coat protein mRNA, into Xenopus laevis oocytes. The two Drosophila isoacceptors are identical besides containing either a guanosine (G) or the hypermodified nucleoside queuosine (Q) in the wobble position. The brewer's yeast isoacceptors differ by four bases in the anticodon stem, and by one base in the amino acceptor stem. Our results show that, under competing 'in vivo' conditions, the Drosophila tRNAHis with the anticodon GUG clearly prefers the histidine codon CAC to the codon CAU, whereas little preference is observed for the tRNAHis with the anticodon QUG for the codon CAU, and no preference for either codon by the two yeast isoacceptors. Hence, it can be concluded that the presence of the Q-base clearly affects the choice of the codon. This is the first demonstration of an 'in vivo' codon preference by tRNA isoacceptors differing in the modification of the wobble base during the elongation step of protein synthesis. These results imply that one function of the Q-base is at the translational level. PMID- 2988937 TI - Electron microscopy of SV40 DNA cross-linked by anti-Z DNA IgG. AB - Electron microscopy has revealed the specific binding of bivalent anti-Z DNA immunoglobulin G (IgG) to different sites on supercoiled Form I SV40 DNA. The anti-Z IgG links together left-handed regions located within individual or on multiple SV40 DNA molecules at the superhelix density obtained upon extraction. Velocity sedimentation, electrophoresis, and electron microscopy all show that two or more Z DNA sites in the SV40 genome can be intermolecularly cross-linked with bivalent IgG into high mol. wt. complexes. The formation and stability of the intermolecular antibody-DNA complexes are dependent on DNA superhelix density, as judged by three criteria: (1) relaxed circular (Form II) DNA does not react; (2) release of torsional stress by intercalation of 0.25 microM ethidium bromide removes the antibody; and (3) linearization with specific restriction endonucleases reverses antibody binding and DNA cross-linking. Non-immune IgG does not bind to negatively supercoiled SV40 Form I DNA, nor are complexes observed in the presence of competitive synthetic polynucleotides constitutively in the left-handed Z conformation; B DNA has no effect. Using various restriction endonucleases, three major sites of anti-Z IgG binding have been mapped by electron microscopy to the 300-bp region containing nucleotide sequences controlling SV40 gene expression. A limited number of minor sites may also exist (at the extracted superhelix density). PMID- 2988938 TI - A low copy number, copia-like transposon in maize. AB - Bs1, a transposable element that moved into the maize Adh1 gene following barley stripe mosaic virus infection, is shown to be present in 1-5 copies in all maize and teosinte lines tested. Bs1 sequences do not hybridize with the genome of barley stripe mosaic virus. The insertion of Bs1 is bounded by 304-bp perfect direct repeats, similar in structure to Ty1 in yeast, copia and related elements in Drosophila, and vertebrate pro-retroviruses, but different from all other known plant transposons. No free copies of the terminal sequences or large internal deletions of Bs elements could be detected. Bs1 is apparently not related to several transposons which moved into the Shrunken gene in lines made genetically unstable by barley stripe mosaic virus infection, suggesting that this virus may cause genome shock, resulting in a generalized liberation of transposons in response to environmental stress. PMID- 2988939 TI - Two classes of fatty acid acylated proteins exist in eukaryotic cells. AB - Labelling of cultured cells with [3H]palmitic and [3H]myristic acids demonstrates that each of these fatty acids modifies a substantially different subset of cellular proteins. Hydroxylamine treatment can be used to differentiate sensitive thioester linkages to palmitate from insensitive amide linkages to myristate. Several palmitoylated proteins are surface-oriented glycoproteins while all of the myristylated proteins appear to be internal. Myristate addition is much more tightly coupled to protein synthesis than palmitoylation, which is able to continue at a reduced level even in the prolonged absence of protein synthesis. Acyl proteins patterns were affected both qualitatively and quantitatively by transformation and growth status. The preferential addition of palmitate to the transferrin receptor and myristate to pp60src, and the absence of these modifications from several other proteins is reported. We propose a nomenclature for fatty acyl proteins based on these observations. PMID- 2988940 TI - Differential Hm antigen expression on EC cells and early differentiated derivatives. AB - Differences in the expression of minor histocompatibility (Hm) alloantigens on two mouse embryonal carcinoma (EC) cell lines and the PYS-2 and T.D.M.-1 differentiated derivatives have been demonstrated by their ability to elicit a cytolytic T-lymphocyte (CTL) response. Experiments involving the use of various responder-target strain combinations on the one hand and Recombinant Inbred (RI) mice strains on the other have shown that: (i) there are major differences in Hm expression on the EC cells compared with the differentiated derivatives whose Hm expression appears more akin to that of adult splenocytes; (ii) although both EC cell lines show reduced Hm immunogenicity compared with adult splenocytes, major differences in the expression and possibly presentation between the F9 and PCC3 EC cell lines can be detected both by in vivo priming and by in vitro cold competition target experiments. These results are discussed in connection with the unexpected finding that some EC cell lines are capable of specific competition effects for appropriate CTL effectors despite their inability to stimulate such effectors in vitro and the absence of major histocompatibility complex (MHC) products. PMID- 2988941 TI - Eukaryotic type I topoisomerase is enriched in the nucleolus and catalytically active on ribosomal DNA. AB - The distribution of eukaryotic DNA topoisomerase I in the cell has been analyzed at four levels: (i) at the level of the nuclear matrix; (ii) at the cytological level by immunofluorescence of whole cells; (iii) at the electron microscopic level using the protein A/colloidal gold technique; and (iv) at the level of DNA to identify in situ the sequence upon which topoisomerase I is catalytically active. Although topoisomerase I is clearly distributed non-randomly in the nucleus, the unique distribution of the enzyme is not related to the nuclear matrix. The data support the conclusion that topoisomerase I is heavily concentrated in the nucleolus of the cell; furthermore, particular regions within the nucleolus are depleted of topoisomerase. A technique has been developed which allows isolation and analysis of the cellular DNA sequences covalently attached to topoisomerase. Ribosomal DNA sequences are at least 20-fold enriched in topoisomerase/DNA complexes isolated directly from a chromosomal setting, relative to total DNA. This is the first direct evidence that topoisomerase I is catalytically active on ribosomal DNA in vivo. PMID- 2988942 TI - A new member of the polyomavirus family: the hamster papovavirus. Complete nucleotide sequence and transformation properties. AB - The hamster papovavirus (HapV) is associated with multiple skin epitheliomas of the Syrian hamster. We have sequenced its genome. It is a double-stranded circular DNA of 5366 bp. The hypothetical genomic organization deduced from this nucleotide sequence is clearly of the polyoma type with the two strands coding in the opposite directions from a noncoding region that shows some of the features of a replication origin and a transcription control region. The amino acid sequences predicted from the open reading frames show an average of 50% homology with polyoma-coded polypeptides. The HapV is, after polyoma, the second example of a papovavirus coding for a middle T antigen. The cloned DNA can immortalize primary rat embryo cells and transform an established rat cell line. The viral DNA is stably integrated into the host genome. PMID- 2988943 TI - Evidence for a repeating domain in type I restriction enzymes. AB - The primary structures of the recognition subunit (hsdS) in type I restriction enzymes from three isolates of Escherichia coli were compared and aligned by use of amino acid physical properties. A repeating domain was found in each of the subunits suggesting a pseudo-dimeric structure. Secondary structure predictions delineated two helical regions in each domain which suggested that the recognition subunits may act in a fashion similar to that proposed for repressor and activator molecules; namely, interaction with double-stranded DNA through helices and in two successive major grooves on the same DNA side. One helical motif could provide the specific recognition site and the other, the restriction site. PMID- 2988944 TI - Use of transposon mutants to assess the role of exoenzyme S in chronic pulmonary disease due to Pseudomonas aeruginosa. AB - Among the potential virulence factors produced by Pseudomonas aeruginosa there are two distinct ADP-ribosyl transferases, exotoxin A and exoenzyme S. The role of exoenzyme S in Pseudomonas aeruginosa infection was studied using the rat chronic pulmonary infection model. Pseudomonas aeruginosa strain DG1 and an isogenic mutant of DG1 differing only in its capacity to produce exoenzyme S were employed in the study. Both Pseudomonas aeruginosa strains tested established a chronic pulmonary infection in this model and organisms recovered from lung homogenates were phenotypically unaltered with respect to exoenzyme S production in vitro. The extent of the observed pathology was markedly greater with the strain producing exoenzyme S, indicating that exoenzyme S may play a role in the progressive pathology observed in chronic lung disease due to Pseudomonas aeruginosa. PMID- 2988945 TI - Role of exoenzyme S in chronic Pseudomonas aeruginosa lung infections. AB - Exoenzyme S is an extracellular ADP-ribosyltransferase enzyme produced by Pseudomonas aeruginosa. Mutants of Pseudomonas aeruginosa deficient in this enzyme have been shown to have reduced virulence in infections of burned mice. The contribution of exoenzyme S to the pathogenesis of chronic lung infections with this organism was evaluated by examining the incidence of exoenzyme S production by Pseudomonas aeruginosa strains isolated from cystic fibrosis patients and comparing an exoenzyme S deficient mutant and its exoenzyme S producing parent in a rat chronic lung infection model. Of 51 isolates examined, 43% produced detectable levels of exoenzyme S. While both the exoenzyme S deficient mutant and its parent strain were equally capable of colonizing and persisting in rat lungs, the exoenzyme S producing parent elicited a greater degree of lung damage. These data suggest that exoenzyme S contributes to the pathogenesis of chronic lung infections. PMID- 2988946 TI - Pulse-chase studies of the synthesis of apolipoprotein B in a human hepatoma cell line, Hep G2. AB - We have used pulse-chase methodology to study the synthesis of apolipoprotein B in a human hepatoma-derived cell line, the Hep G2 cells. A 2-min pulse with [35S]methionine was followed by a chase period varying from 5-90 min. A protein of large molecular mass (estimated molecular mass: 312 +/- 41 kDa, mean +/- SD, n = 8) could be immunoprecipitated from the cells at all chase periods between 5 min and 60 min with both monoclonal antibodies to a narrow density cut of the low density lipoprotein LDL-2 (density: 1.030-1.055 g/ml) and polyclonal antibodies to the apolipoprotein B apo B 100 or to a narrow density cut of LDL-2 (density: 1.030-1.055 g/ml). In addition to this large molecular mass protein, nascent polypeptides could be precipitated after 5, 10 and 15 min of chase. The apolipoprotein B molecules that had been labelled during the pulse disappeared from the cells after 60-90 min of chase, while they started to appear in the medium after 30-35 min of chase. The results obtained indicate (a) that apolipoprotein B is synthesized as one polypeptide with a large molecular mass, (b) that newly synthesized apolipoprotein B molecules are secreted after a delay of 30-35 min, (c) that no intracellular accumulation of apolipoprotein B occurs, and (d) that apolipoprotein B is recovered in the density fraction less than 1.21 g/ml of the medium suggesting that it is secreted in lipoprotein form. PMID- 2988947 TI - Glycinin A5A4B3 mRNA: cDNA cloning and nucleotide sequencing of a splitting storage protein subunit of soybean. AB - The complete nucleotide sequence of a cloned cDNA, designated pGA5A4B3822, corresponding to glycinin A5A4B3 mRNA was determined. Analysis of the cDNA insert revealed that it contained 1899 nucleotides of mRNA sequence with a 5'-terminal non-translated region of 31 nucleotides, a signal peptide region corresponding to 23 amino acids, an acidic subunit region (A5) corresponding to 97 amino acids, an acidic subunit region (A4) corresponding to 257 amino acids followed by a basic subunit region (B3) corresponding to 185 amino acids, and a 3'-terminal non translated region of 182 nucleotides. These results show that the glycinin A4 subunit, which is not found to be linked to a basic subunit via a disulfide bond, is synthesized as a full-sized precursor, i.e. the A5A4B3 subunit complex, from a single mRNA, followed by post-translational processing to generate an intermediary subunit complex (A5-B3), covalently linked by a disulfide bond, and the mature A4 subunit, which may associate with the above subunit complex by non covalent interactions. From the results obtained by the Chou-Fasman rules we speculated that the two post-translational cleavage sites of this subunit precursor might be processed by the same proteolytic enzyme. PMID- 2988948 TI - Cooperation among electron-transfer complexes in ATP synthesis in chloroplasts. AB - We have investigated the extent to which redox reactions of thylakoid membranes cooperate in ATP synthesis. This was done by measuring the onset of ATP synthesis following a series of single-turnover light-flashes at various levels of electron transport inhibition. In the presence of antibiotics that prevent the formation of a membrane potential, the onset of ATP synthesis seems to depend entirely on the formation of an adequately large delta pH. Under our conditions, the accumulation of about 60 mmol H+ X mol chlorophyll-1 is then necessary to form the requisite delta pH, which in turn requires about 15 saturating flashes in uninhibited thylakoid samples. Inhibition of some of the electron transfer centers by limiting the light intensity of the flashes, by dichlorophenyldimethylurea, by heat treatment, or by NH2OH-treatment caused an increase in the number of flashes required for the onset of ATP synthesis. The increase in the requisite number of flashes reflected the decreased number of electrons transferred in each flash, almost exactly the same number of electrons being transferred before ATP synthesis could begin. This effect of inhibitors was true when the two photosystems were operating in unison and when either of the two photosystems was acting alone. However, when either photosystem acted alone, there was an increase in the number of flashes required for the onset of ATP synthesis, an increase which was consistent with the observed lower flash-induced proton accumulation. A mathematical analysis of the onset of flash-induced ATP synthesis shows that at least several hundred proton-translocating electron transport complexes must be cooperating to form the threshold delta pH. In spite of this evidence for extensive cooperation among different electron transport complexes in ATP formation, the implied pooling of H+ ions does not seem to involve inner vesicle regions accessible to exogenous buffers. Thus, even when the number of H+ ions accumulated per flash was reduced by 70% through attenuation of the intensity of the flashes, exogenous hydrogen ion buffers present within the lumen of the thylakoid vesicle had no effect on the number of flashes required for the onset of ATP synthesis. PMID- 2988949 TI - Protonmotive force and photophosphorylation in single swollen thylakoid vesicles. AB - Swollen vesicles generally 40 micron in diameter were prepared from spinach chloroplasts. These vesicles appear to originate from thylakoids. The present study reports results obtained with individual vesicles using micromanipulative procedures. The electric potential across the membrane was measured with microelectrodes and the pH of the internal space was calculated from the fluorescence of the pH indicator pyranine. The individual vesicles photophosphorylate as measured with luciferin-luciferase. Impalement with microelectrodes did not affect the ability of individual vesicles to photophosphorylate. However, there was no significant membrane potential either with continuous illumination or light flashes. In contrast, we found a delta pH of 3.7 under photophosphorylative conditions and the incubation with the appropriate buffers blocked photophosphorylation presumably by preventing formation of a pH gradient. We propose that, in these vesicles, the membrane potential plays no role in photophosphorylation, whereas a pH gradient is obligatory. PMID- 2988950 TI - Heterogeneous glycosylation of murine transferrin receptor subunits. AB - The N-linked glycosylation of the murine receptor for transferrin has been investigated. Previously we have found that purified receptors appear as two bands after sodium dodecyl sulfate/polyacrylamide gel electrophoresis and Coomassie blue staining [van Driel, I.R., Stearne, P.A., Grego, B., Simpson, R.J. and Goding, J.W. (1984) J. Immunol. 133, 3220-3224]. In the current report we show that the two bands are due to different glycosylation of individual receptor molecules. The receptors have three asparagines to which N-linked glycans can be added, but only two sites are glycosylated in all receptors. The level of glycosylation of the third site varies depending on cell line or tissue. Limited endoglycosidase digestion of mature receptors indicates that differential glycosylation probably occurs at only one particular asparagine residue. Possible mechanisms that could result in such a glycosylation pattern are discussed. PMID- 2988951 TI - The complete 30-base-pair origin region of bacteriophage phi X174 in a plasmid is both required and sufficient for in vivo rolling-circle DNA replication and packaging. AB - The origin of replication of the isometric single-stranded DNA bacteriophages is located in a specific sequence of 30 nucleotides, the origin region, which is highly conserved in these phage genomes. Plasmids harboring this origin region are subject to rolling-circle DNA replication and packaging of single-stranded (ss) plasmid DNA into phage coats in phi X174 or G4-phage-infected cells. This system was used to study the nucleotide sequence requirements for rolling-circle DNA replication and DNA packaging employing plasmids which contain the first 24, 25, 26, 27, 28 and the complete 30-base-pair (bp) origin region of phi X174. No difference in plasmid ss DNA packaging was observed for plasmids carrying only the 30-bp origin region and plasmids carrying the 30-bp origin region plus surrounding sequences (i.e. plasmids carrying the HaeIII restriction fragment Z6B of phi X174 replicative-form DNA). This indicates that all signals for DNA replication and phage morphogenesis are contained in the 30-bp origin region and that no contribution is made by sequences which immediately surround the origin region in the phi X174 genome. The efficiency of packaging of plasmid ssDNA for plasmids containing deletions in the right part of the origin region decreases drastically when compared with the plasmid containing the complete 30-bp origin region (for a plasmid carrying the first 28 bp of the origin region to approximately 5% and 0.5% in the phi X174 and G4 systems respectively). Previous studies [Fluit, A.C., Baas, P.D., van Boom, J.H., Veeneman, G.H. and Jansz, H.S. (1984) Nucleic Acids Res. 12, 6443--6454] have shown that the presence of the first 27 bp of the origin region is necessary as well as sufficient for cleavage of the viral strand in the origin region by phi X174 gene A protein. Moreover, Brown et al. [Brown, D.R., Schmidt-Glenewinkel, T., Reinberg, D. and Hurwitz, J. (1983) J. Biol. Chem. 258, 8402--8412] have shown that omission of the last 2 bp of the origin region does not interfere with phi X174 rolling-circle DNA replication in vitro. Our results therefore suggest that for optimal phage development in vivo, signals in the origin region are utilized which have not yet been noticed by the in vitro systems for phi X174 phage DNA replication and morphogenesis. PMID- 2988952 TI - Chemical modification of methionine residues of the phosphatidylcholine transfer protein from bovine liver. A spin-label study. AB - The role of methionine residues in the interaction of the phosphatidylcholine transfer protein from bovine liver with phospholipid vesicles was investigated by specific modification of these residues with iodoacetamide. The modified protein was digested with cyanogen bromide in order to determine which methionine residues had become resistant to this cleavage. Automated Edman degradation on the digest indicated that after 72 h of reaction, Met-1 was modified for 80%, Met 73 for 50%, Met-109 for 20%, whilst Met-173 and Met-203 were found to be unmodified. This distinct modification did not result in any loss of phosphatidylcholine transfer activity. The interaction of the phosphatidylcholine transfer protein with phospholipid vesicles was investigated by making use of electron spin resonance spectroscopy. The interaction of unmodified protein with vesicles composed of phosphatidylcholine/phosphatidic acid/spin-labeled phosphatidylethanolamine (79:16:5, mol%) or composed of phosphatidylserine/spin labeled phosphatidylethanolamine (95:5, mol%), gave an increase of about 50% in the rotation correlation time. A similar increase was observed with the modified protein. This interaction was further investigated by labeling Met-1 and Met-73 in the transfer protein with iodoacetamidoproxyl spin-label. Spin-labeling did not inactivate the transfer protein. In addition, the electron spin resonance spectra of the spin-labeled protein were not affected upon addition of vesicles composed of phosphatidylcholine/phosphatidic acid (80:20, mol%). These experiments strongly suggest that Met-1 and Met-73 are not part of the site that interacts with the membrane. PMID- 2988953 TI - Natural-abundance 13C nuclear magnetic resonance studies of regulation and overproduction of L-lysine by Brevibacterium flavum. AB - Natural-abundance 13C NMR spectroscopy has been used to study the metabolism of the L-lysine-producing bacterium, Brevibacterium flavum. Relationships of biomass formation, precursor uptake, and product excretion, as a function of culture medium, oxygen supply and specific cell membrane permeability, were rapidly determined using 67.89-MHz 13C NMR. The induction of lysine production throughout the growth cycle was studied. Intracellular and extracellular levels of free metabolites and unconsumed precursor were quantitatively measured as a function of growth culture conditions. Limited availability of oxygen resulted in accumulation and excretion of unfavorable products: lactate, succinate, alanine and valine. However, under optimal aeration conditions L-lysine was the sole metabolite detected extracellularly. Various important long-lived intermediates and storage compounds were detected in the intact cells (by NMR measurements). Carbon resonances of carbohydrates and amino acids were resolved and easily identified. Of particular interest are those of trehalose carbons, a storage carbohydrate. Natural-abundance 13C NMR spectroscopy seems most suitable for biotechnological processes where high concentrations of intermediates and end products can be observed. We anticipate that this approach will be employed to screen overproducing bacterial strains. PMID- 2988954 TI - Quantitative and mechanistic aspects of the hydroperoxide-induced release of Ca2+ from rat liver mitochondria. AB - We have previously demonstrated in rat liver mitochondria a hydroperoxide-induced hydrolysis of pyridine nucleotides and release of Ca2+ [Lotscher, H. R., Winterhalter, K. H., Carafoli, E. & Richter, C. (1979) Proc. Natl Acad. Sci. USA 76, 4340-4344, and Lotscher, H. R., Winterhalter, K. H., Carafoli, E. & Richter, C. (1980) J. Biol. Chem. 255, 9325-9330]. Here we investigate pyridine nucleotide hydrolysis and Ca2+ release under conditions of minimized Ca2+ cycling and with smaller Ca2+ loads. The extent of pyridine nucleotide hydrolysis, measured by pyridine-nucleotide-derived nicotinamide release from intact mitochondria, and the Ca2+ release rate show a very similar sigmoidal dependence on the mitochondrial Ca2+ load. The hydrolysis of oxidized pyridine nucleotides is limited under non-cycling conditions. Whereas pyridine nucleotide hydrolysis as measured by nicotinamide release is extensive, net loss of mitochondrial pyridine nucleotides is observed only at relatively high Ca2+ loads. Our results indicate the ability of mitochondria to resynthesize pyridine nucleotides after hydrolysis. Neither a decrease of reduced, nor an increase of oxidized, mitochondrial glutathione favour Ca2+ release. From these and previous findings it is concluded that the hydroperoxide-induced Ca2+ release is triggered by a factor which is distal to the oxidation of mitochondrial pyridine nucleotides. Ca2+ release is stimulated when the movement of protons across the inner mitochondrial membrane is facilitated, giving evidence for the operation of the hydroperoxide-induced release pathway as a Ca2+/H+ antiport. PMID- 2988955 TI - Imino-proton resonances of yeast tRNAPhe studied by two-dimensional nuclear Overhauser enhancement spectroscopy. AB - Application of two-dimensional nuclear Overhauser enhancement (NOE) spectroscopy to yeast tRNAPhe in H2O solution demonstrates that all imino-proton resonances, related to the secondary structure, and nearly all imino proton resonances, originating from the tertiary structure, can be assigned efficiently by this method. The results corroborate the assignments of the imino-proton resonances of this tRNA as established previously by one-dimensional NOE experiments (only the assignment of base pairs G1 X C72 and C2 X G71 should be reversed). The advantages of two-dimensional NOE spectroscopy over one-dimensional NOE spectroscopy for the assignments of imino-proton resonances and the structure elucidation of tRNA are illustrated and discussed. Furthermore, the use of non exchangeable proton resonances as probes of the molecular structure is explored. PMID- 2988956 TI - Lateral proton conduction at a lipid/water interface. Its modulation by physical parameters. Experimental and mathematical approaches. AB - Fast lateral proton conduction along the lipid/water interface has recently been experimentally demonstrated in our laboratory [Teissie, J., Prats, M., Soucaille, P. & Tocanne, J.F. (1985) Proc. Natl Acad. Sci. USA, in the press]. The present study gives a more precise description of the way various physical parameters can affect this process. The dependence of the distance covered by the proton on time is demonstrated to be quadratic. Increasing the speed of stirring in the injection compartment or the amount of injected acid or the contact between the monolayer and the acidic subphase increased the efficiency of the proton transfer. Raising the strength of the buffer in the bulk phase inhibited proton conduction. Results from experiments where the transfer of protons from the bulk phase to the interface was modified, suggested the occurrence of an 'energy barrier' limiting the access of protons from the bulk phase to the lipid polar head region. PMID- 2988957 TI - Preoperative scintigraphic evaluation of the location of juxtaglomerular cell tumor. AB - Juxtaglomerular cell tumor is a rare condition which cannot be correctly diagnosed preoperatively. In the only previously reported case in which scintigraphy has been used, this method failed to detect the tumor. We present a case of juxtaglomerular cell tumor in which renal scintigraphy revealed a well defined cold area in the hilar region of the left kidney. PMID- 2988958 TI - The 15O continuous-inhalation method: correction for intravascular signal using C15O. AB - Cerebral oxygen metabolism (CMRO2) and fractional extraction (OEF) regional values obtained using the 15O steady-state method and positron-emission tomography (PET) were corrected for the activity remaining in blood vessels (a source of overestimation of the OEF neglected in the simple steady-state model) using an additional scan during continuous inhalation of nontoxic amounts of carbon monoxide labeled with 15O (C15O). The method was applied to ten serial PET studies. In normal tissue, OEF overestimation was 11% +/- 2.6% and 9% +/- 2.0% in gray and white matter, respectively. In pathological tissue, the OEF overestimation was, on average, higher than in normal tissue (28% +/- 17% in the core of lesions), but more variable. In both normal and abnormal tissue, however, it was found that (1) the lower the real OEF, the larger the correction applied, and (2) the correction appears to be particularly necessary in situations of abnormally low OEF. The use of C15O continuous inhalation is a simple, direct, and accurate method for blood-activity correction when the 15O steady-state technique is employed to study pathophysiology. PMID- 2988959 TI - Scintigraphic detection of adenoma in primary hyperparathyroidism. AB - Ten patients with biochemical and clinical evidence of primary hyperparathyroidism were studied scintigraphically using a double-radionuclide subtraction technique. Preoperative imaging was performed with Sodium pertechnetate Tc 99m and thallium chloride Tl 201 using a gamma camera with a pinhole collimator. The data were stored in a matrix of 64 X 64. The avidity of 201Tl for nodules of the parathyroid gland was demonstrated on analogue and digital images both with and without the subtraction of thyroid tissue. In our ten patients, this technique detected nine out of ten histologically proven adenomas, and missed a relatively small lesion embedded within the thyroid of one patient. This method may have considerable advantages over non-specific or invasive radiodiagnostic methods, and may facilitate the diagnosis and planning of the surgical approach in cases of primary hyperparathyroidism. PMID- 2988961 TI - Radio-synovectomy in chronic synovitis of the knee joint in patients with rheumatoid arthritis. AB - The influence of intra-articular (i.a.) colloidal 198Au (5 mCi) or 90Y-silicate (5 mCi) on synovitis of the knee joint in patients older than 45 years with rheumatoid arthritis (RA), who had been treated since 1970 in our hospital, was investigated. Of the 89 knee joints of 77 patients studied, 65 had no or minimal radiological abnormalities of the knee joint treated (group I), whereas 24 patients had moderate to severe changes (group II). Before and at regular intervals after radio-synovectomy the clinical response was scored using pain, hydrops and warmth as parameters. The results indicated that 1 year after treatment the percentage of knee joints with a favourable response was greater in group I than in group II (58% versus 25%, P = 0.001). This difference was still present 3 years after treatment. Clinical response showed no correlation with initial inflammatory activity as measured by 99mTc-pertechnetate uptake measurements. However, in group I, those patients with an ESR below 60 mm/h, measured just before radio-synovectomy, more often had a favourable response than those with an ESR in excess of 60 mm/h (P = 0.01). No or only slight complications of radio-synovectomy were noted, whereas leakage of radioactivity from the knee joints was minimal. It is concluded that radio-synovectomy is an effective and safe procedure in those patients with rheumatoid synovitis of the knee joint without the presence of significant radiological damage and the absence of active systemic disease. PMID- 2988962 TI - False-positive Meckel's scan in a case of infected ovarian cyst. AB - Abdominal scanning using sodium pertechnetate Tc 99m is useful in demonstrating Meckel's diverticula containing ectopic gastric mucosa. However, other structures like the uterus may also concentrate 99mTc. We present a case which showed a focal area of increased uptake in the pelvis due to an infected ovarian cyst. PMID- 2988960 TI - Measurement of cerebral circulation time in man. AB - A simple, inexpensive method for measuring the cerebral circulation time (CCT) was developed. The CCT was considered to be equal to the time that an intravenously injected bolus of sodium pertechnetate Tc 99m took to go from the subclavian artery to the posterior venous confluence. The dilution curves were externally recorded at these two vessels. Particular attention was given to the curve treatment. The computer programmes were specially conceived in order to detect and, if possible, correct the experimental curve defects. Several reliability criteria were also defined in order to test the validity of each measurement. From a physiological point of view, the CCT is equal to the ratio of the global cerebral blood volume to the global cerebral blood flow. Thus, it can be widely applied in clinical research. For example, in a study of the relationship between CCT and age, sex and light cerebrovascular impairment, the technique was sensitive enough to discriminate between healthy subjects and patients suffering from-transient ischemic attack, or regressive stroke, and to show the action of a drug on the cerebral circulation in such patients. PMID- 2988963 TI - Delayed contralateral hemiplegia following herpes zoster ophthalmicus: should antiviral therapy be used? AB - We review clinical virological studies in the syndrome of delayed contralateral hemiplegia following herpes zoster ophthalmicus. Virus could not be isolated from the cerebrospinal fluid (CSF) of the present case, nor was antiviral antibody found in the CSF. There appear to have been no reports of successful virus isolation from the CSF although there are reports of antibody in the spinal fluid. Thus the evidence for ongoing viral replication in the central nervous system is marginal. It is suggested that the sensitive antibody assay against membrane antigens (FAMA) be used in the future as a guide to antiviral therapy. PMID- 2988964 TI - Morphological studies of peripheral nerves and skeletal muscles of an adult case with adrenoleukomyeloneuropathy. AB - A 53-year-old patient with adrenoleukomyeloneuropathy, showing symptoms of mental deterioration, cerebellar ataxia, spastic tetraparesis, and polyneuropathy, was reported. The myelinated fiber densities were decreased in the anterior spinal root and sciatic nerve. Most of the myelinated fibers showed segmental demyelination and a few fibers showed axonal degeneration. Electron microscopy showed numerous lipid inclusions in the cytoplasm of the Schwann cells. The muscles showed neurogenic changes. Numerous lipid bodies and intracytoplasmic crystalline arrays were found in the sarcolemma. PMID- 2988966 TI - Pulmonary function to measure response in small cell lung cancer. PMID- 2988965 TI - Relapse pattern of pathologic stage I nonseminomatous germ cell tumors of the testis following orchidectomy and lymphadenectomy. AB - A retrospective analysis of patients with pathologic stage I nonseminomatous germinal testis tumors treated with orchidectomy and retroperitoneal lymphadenectomy was done in an attempt to define the relapse pattern and to eventually identify risk factors predisposing to tumor recurrence. Of 102 patients, 91 (89.2%) remain disease free with a median follow-up of greater than 5 years (range 3-10 years), and 11 (10.8%) suffered relapse 3-35 months after lymphadenectomy (median free interval 6 months). 9 of 11 patients developed primarily intrathoracic recurrences. The tumor was so rapidly progressing at relapse, that it was diagnosed when clinically advanced in 7 of 11 cases. Nevertheless, 8 of 11 patients were salvaged with effective available chemotherapy and resection of residual disease. Only scrotal violation showed a significant increased risk of tumor recurrence (5 of 24 cases, versus 6 of 78 who had inguinal orchidectomy, p less than 0.05). We recommend a very close follow-up for all patients with pathologically assessed stage I nonseminomatous germinal testis tumors during the first 3 years following orchidectomy and retroperitoneal lymphadenectomy. With early recognition of relapse, an approximately 100% cure rate will be achieved with effective available chemotherapy. PMID- 2988967 TI - Combination endocrine therapy in breast cancer. PMID- 2988968 TI - Effects of cytostatic drugs and 40.5 degrees C hyperthermia on human clonogenic tumor cells. AB - A tumor colony-forming assay was used to investigate the effect of 40.5 degrees C hyperthermia and drugs on the colony-forming ability of human clonogenic tumor cells. In order to be able to perform repeated incubations with identical tumor material, specimens were used that were augmented in the nude mouse system. Five tumor samples (two malignant melanomas, two squamous cell carcinomas of the lung and one small cell carcinoma of the lung) were incubated with seven drugs: doxorubicin, actinomycin-D, bleomycin, melphalan, vincristine, vinblastin and cisplatinum. One additional tumor specimen (chondrosarcoma) was incubated with doxorubicin immediately after resection. The incubation with drugs at 37 degrees C revealed dose response curves typical for each tumor. Incubation at 40.5 degrees C for 2 hr showed enhanced drug effects in five out of the six tumors tested. The drugs, with an enhanced effect and the pattern of enhancement were different in each individual tumor. PMID- 2988969 TI - Changing histology in malignant tumors: diagnostic and therapeutic significance. PMID- 2988970 TI - Polyoma virus-induced hemangiomas in grafts of visceral yolk sac and of embryos. AB - The development of hemangiomas in grafts of visceral yolk sac and embryos after in vitro infection with polyoma virus is described. These hemangiomas were shown to be positive for the polyoma tumor-specific transplantation antigen(s) and to develop as well in recipient virgin as in pregnant rats. In grafts of fetal skin or gut infected with polyoma virus neoplastic transformation was never observed. PMID- 2988971 TI - Regulation of hepatic glucose production during exercise. PMID- 2988972 TI - Haemodynamic and pharmacological effects of the converting enzyme inhibitor CGS 14824A in normal volunteers. AB - The converting enzyme inhibitor CGS 14824A was evaluated in 15 healthy male volunteers. First, the efficacy of a single 5 or 10 mg oral dose in antagonizing the pressor response to exogenous angiotensin I was tested in 2 subjects. Blood pressure and heart rate were monitored continuously through an intra-arterial catheter. CGS 14824A 5 mg reduced the response to angiotensin I within 75 min to 50%, and 10 mg within 1 h to less than 25%, and for a period of more than 4 h. Subsequently, plasma renin and converting enzyme activity, plasma angiotensin I, angiotensin II and aldosterone were measured serially before and up to 72 h following oral administration of 2, 5, 10 or 20 mg CGS 14824A to groups of 5 volunteers. Plasma converting enzyme activity fell to well below 10% of baseline within 1 h after administration of 5 mg or more CGS 14824A. Within 2 h following 2 mg p.o., a similarly low level was reached. Twenty four hours following the 20 mg dose, plasma converting enzyme activity was still below 10%. As expected, plasma renin activity and angiotensin I rose while angiotensin II and aldosterone fell following the 2 mg dose. This pattern of effects was enhanced by increasing the dose. Nonetheless, 24 h after the 20 mg dose, plasma angiotensin II and aldosterone had returned to their baseline levels. No side-effects occurred. Thus, in normal volunteers, CGS 14824A was an effective, potent and long acting converting enzyme inhibitor. PMID- 2988973 TI - The respiratory burst is not required for killing of intracellular and extracellular parasites by a lymphokine-activated macrophage cell line. AB - The macrophage cell line, IC-21, was found to be incapable of producing the oxygen products associated with the respiratory burst. However, IC-21 cells were activated by lymphokine (LK) to kill intracellular (Leishmania donovani amastigotes) and extracellular (Schistosoma mansoni larvae) parasites, as well as tumor cells. In each case, the cytotoxicity exhibited by activated IC-21 cells and activated peritoneal macrophages was indistinguishable. However, nonactivated IC-21 cells were unable to kill L. donovani log-growth phase promastigotes, while nonactivated peritoneal macrophages destroyed greater than 90% of the initial infection. These results indicate that amastigotes and schistosome larvae are susceptible to killing by nonoxidative cytotoxic mechanism induced by lymphokine activation but, on the other hand, support the concept that the killing of log growth phase promastigotes by nonactivated cells is dependent upon the respiratory burst. We propose that the IC-21 cell line may be a useful model for studying nonoxidative killing functions of activated macrophages. PMID- 2988974 TI - Isolation and characterization of an Fc receptor from neonatal rat small intestine. AB - Receptors for the Fc region of IgG from neonatal rat intestinal brush borders were solubilized using 3-[(3-cholamidopropyl)dimethyl-ammonio]-1-propane sulfonate and purified by affinity chromatography. Analysis of IgG-binding material by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions reveals two components with apparent Mr of 41 000-50 000 and 15 000. The larger component is glycosylated and may dimerize, giving a 100-110 kDa band on nonreduced gels. Both proteins are localized in the proximal small intestine, where IgG is specifically taken up during the first three weeks of neonatal life, and disappear when specific transport stops after weaning. Electron irradiation of brush borders shows that the functional unit for IgG binding has a molecular weight in situ of 110 kDa. These data suggest that a dimer of the 41-50-kDa protein together with the 15 kDa and other proteins may mediate intestinal transport of maternal IgG. PMID- 2988975 TI - Adenosine increases an internal calcium store in the smooth muscle of guinea-pig taenia coli. AB - The effect of adenosine on an internal calcium store which can be released by carbachol to produce a transient contraction in Ca2+-free solution was investigated in the taenia depolarized by high K+. The carbachol contraction in Ca2+-free solution was increased by the preceding application of adenosine (3 X 10(-5) to 10(-3) M), an effect which was mimicked by ATP, but not by the slowly degradable analog alpha, beta-methylene ATP. The P1-purinoceptor antagonist 8 phenyltheophylline inhibited the increase caused by adenosine without modifying the carbachol contraction in controls. It is concluded that stimulation of an extracellular P1-purinoceptor increases an internal store of calcium, which might contribute to the relaxation induced by adenosine in the taenia. PMID- 2988976 TI - Topography of binding sites for neurohypophyseal hormones in rat brain. AB - Binding sites for arginine-vasopressin (AVP) and oxytocin (OT) were detected in neuroanatomically defined regions of rat brain by using in vitro incubation of 32 micron brain sections with the tritium-labelled ligands and autoradiography with LKB Ultrofilm. Sites were quantified by computerized densitometry of the film images. The highest amount of [3H]AVP-labelled binding sites were observed in the ventral subiculum of the hippocampus, central amygdala nucleus, lateral septum, olfactory nucleus and nucleus tractus solitarii, while the dentate gyrus contained a large amount of low affinity binding. The binding pattern of [3H]OT was more discrete. Relatively high concentrations of OT binding sites were observed in the central amygdala, ventral subiculum and olfactory nucleus. The lateral septum and the nucleus tractus solitarii were virtually devoid of displaceable OT binding while no significant association of OT with the dentate gyrus was found. The topography of binding sites for neurohypophyseal hormones corresponds to the localization of responsive sites in behavioural, biochemical and electrophysiological studies. PMID- 2988977 TI - Pindolol decreases plasma angiotensin-converting enzyme activity in young spontaneously hypertensive rats. AB - The beta-adrenoceptor antagonist pindolol, given p.o. (10 mg/kg, once a day for 7 days) to 4 and 24 week old male spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY), decreased plasma angiotensin-converting enzyme (ACE) activity only in young SHR. Kinetic studies indicated that the treatment reduced the ACE maximal velocity or the number of available ACE molecules. In vitro studies showed that pindolol had no direct effects on the activity of plasma ACE. A decrease in soluble (loosely bound) ACE activity was observed in the lung of young SHR treated with pindolol. The results suggest that pindolol affects the site of origin of plasma ACE. The susceptibility of young SHR to pindolol may indicate a probable role of beta-adrenoceptors in the regulation of ACE activity in spontaneous (genetic) hypertension. PMID- 2988978 TI - Why is amitriptyline much weaker than desipramine at decreasing beta-adrenoceptor numbers? AB - Desipramine is consistently more effective than amitriptyline at causing beta adrenoceptor down-regulation. Atropine, mepyramine, ketanserin, cyproheptadine and citalopram did not modify this action of desipramine in rats. Therefore inhibition of either muscarinic, histamine-H1, and 5-HT receptors or 5-HT uptake produced by amitriptyline is unlikely to account for its weaker effect on beta adrenoceptors. A more likely explanation implicates noradrenaline uptake inhibition in vivo since amitriptyline was much weaker than desipramine and only effective after repeated dosing. PMID- 2988979 TI - Adrenalectomy reduces the density of 'peripheral-type' binding sites for benzodiazepines in the rat kidney. PMID- 2988980 TI - Relationship between pituitary responses to human corticotropin-releasing factor and thyrotropin-releasing hormone in depressives and normal controls. PMID- 2988981 TI - Depletion of peripheral-type benzodiazepine receptors after hypophysectomy in rat adrenal gland and testis. AB - We examined the effect of hypophysectomy in rats on peripheral-type benzodiazepine receptors in the adrenal gland and testis. Following removal of the pituitary, binding sites for [3H]Ro5-4864, a ligand selective for peripheral type benzodiazepine receptors, are depleted in these tissues in parallel with their atrophy. This reduction in receptor levels appears to be a primary effect of the hypophysectomy, since castration has no effect on benzodiazepine receptors in the adrenal gland, nor does adrenalectomy influence testicular levels of these receptors. Our results indicate that peripheral-type benzodiazepine receptors in these steroidogenic organs are localized to cells which are dependent on trophic maintenance by the pituitary. PMID- 2988982 TI - Profile of activity of kappa receptor agonists in the rabbit vas deferens. AB - The purpose of this study was to investigate further the kappa opioid receptor selectivity of the field-stimulated isolated rabbit vas deferens preparation and to study the profile of a series of kappa agonists in this tissue. Agonists acting at mu, delta and sigma receptors were without detectable effect in the rabbit vas deferens. But a number of kappa agonists, including bremazocine, tifluadom, ethylketocyclazocine, ketocyclazocine, U-50,488 and Win 42,610 all depressed contractions, producing parallel dose-response curves. Mr 2034 generally produced a shallower dose-response curve and achieved a lower maximum effect, thus acting like a partial agonist. The effect of ethylketocyclazocine was not reduced by the irreversible mu antagonist, beta-funaltrexamine, confirming that it is not acting via mu receptors. Another group of drugs, including nalorphine, butorphanol and proxorphan, which produce an agonist action via kappa receptors in the guinea-pig ileum and mouse vas deferens, were antagonists in the rabbit vas deferens, suggesting that this tissue will only respond to high efficacy kappa agonists. PMID- 2988984 TI - Opioid dependence in the guinea-pig myenteric plexus is controlled by non tolerant and tolerant opioid receptors. AB - The experiments concerned the association of opioid dependence with specific opioid receptors. Previous investigations have demonstrated that tolerance may be confined to only one type of opioid receptor. These findings could suggest that dependence develops invariably only with receptors which have been rendered tolerant. We report here that in the guinea-pig isolated ileum, which has been made selectively tolerant to a mu-receptor agonist, naloxone may precipitate a sign of dependence at mu-receptors chronically activated and at naive kappa receptors. Furthermore, suppression of the withdrawal contracture in preparations rendered dependent on a mu-receptor agonist can be achieved by very low concentrations of a kappa-agonist, e.g. dynorphin A. These findings challenge the current concept that confines drug dependence only to that opioid receptor type which has been activated chronically. PMID- 2988983 TI - Potentiation of the inhibitory effect of a thromboxane A2 antagonist (L-640,035) on arterial thrombosis formation in rabbit by the angiotensin converting enzyme inhibitor enalapril. AB - The angiotensin converting enzyme inhibitor enalapril (0.5 mg/kg i.v.) potentiated significantly the inhibitory effect of the thromboxane A2 antagonist L-640,035 (1 mg/kg i.v.) on electrically induced platelet accumulation in the rabbit in vivo. Enalapril had no effect upon platelet accumulation when given alone. The hypotensive effects of enalapril did not account for the potentiation because a combination of hexamethonium (5 mg/kg i.v.) and hydralazine (1 mg/kg i.v.), which decreased blood pressure similarly to enalapril, did not augment the effect of L-640,035. Determination by radioimmunoassay of plasma levels of immunoreactive 6-keto-PGF1 alpha, suggested that increases in PGI2 levels after combined administration of enalapril and L-640,035 could explain the observed potentiation. PMID- 2988985 TI - Arachidonic acid, an in vivo inhibitor of carrageenin induced granulomas in the rat. AB - Arachidonic acid (AA) injected locally into carrageenin/sponge granulomas, but not if given orally, inhibited granuloma growth. Granuloma macrophage (M0) infiltration was inhibited, and prostaglandin E2 (PGE2) synthesis (ng/100 mg granuloma dry weight) stimulated, by AA treatment. M0 adenosine 3',5'-cyclic monophosphate (cAMP) levels and granuloma exudate volume were not affected. Granuloma M0s incubated in vitro with arachidonic acid synthesised thromboxane B2 (Txb2), 6-ketoprostaglandin F1 alpha (6-ketoPGF1 alpha), and preferentially, PGE2. The AA inhibition of granuloma growth was possibly mediated via the synthesis of PGE2. PMID- 2988986 TI - Age-related changes in 5HT uptake and [3H]imipramine binding sites in rat cerebral cortex. PMID- 2988987 TI - Evidence for the existence of an actin-derived protein in ataxia-telangiectasia lymphoblastoid cell lines. AB - In a number of lymphoblastoid cell lines from individuals with the genetic disease ataxia-telangiectasia (A-T), a decrease in the levels of actin and a concomitant increase in the levels of a protein species of 37K has been observed to occur following high cell density. In this paper we describe ataxia telangiectasia lymphoblastoid lines that show this response constitutively, and demonstrate that this 37K protein is most likely an actin-derived peptide. This is because of extensive similarity between the two proteins in the spectrum of peptides produced by proteolytic fingerprinting. It appears likely that this 37K protein is similar to the well documented protease-resistant actin fragments produced by protease digestion of actin in vitro. Examination of a number of possible initiators found no clear signal that promoted this response, although the response is clearly very dependent on cell culture conditions. The possible involvement of a cytoskeletal anomaly in ataxia-telangiectasia is discussed. PMID- 2988988 TI - Focal contacts of normal and RSV-transformed quail cells. Hypothesis of the transformation-induced deficient maturation of focal contacts. AB - Morphology and distribution of cell-substrate contacts and their association with microfilament bundles in normal and RSV-transformed quail fibroblasts (16Q line) were studied by indirect immunofluorescence. The focal contacts were visualized by antibody exclusion method using monoclonal antibody to 80 kD serum protein adsorbed on the substratum. Embryo quail cells formed focal contacts of two morphological types: (1) small punctate; and (2) elongated large contacts. These two variants of contacts were designated respectively as dot and dash contacts. Both of focal contacts contained vinculin and alpha-actinin. Double immunofluorescence staining with polyclonal antibody to actin and monoclonal antibody to vinculin revealed that the dot contacts, in contrast to the dash ones, were not associated with microfilament bundles. The dot contacts were localized mostly near the active cell edges, while the dash contacts were found near the retracted cell edges and also under the central parts of the cell. We suppose that dot contacts represent initial structures which then can undergo maturation transforming them into dash contacts. RSV-transformed 16Q cells had predominantly the dot contacts which were not only located at the edges but also in the more central parts of the lamella. The dash contacts were present only in the minority of 16Q cells. RSV transformation is assumed to affect not the ability of cells to form initial dot contacts, but the maturation of dot contacts into dash contacts. PMID- 2988989 TI - Responses of the pigmented rabbit retina to NMPTP, a chemical inducer of parkinsonism. AB - The electrophysiological and neurochemical effects of N-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (NMPTP), a chemical inducer of Parkinsonism in man and monkeys, on the pigmented rabbit retina were determined under both chronic and acute conditions. The implicit time, the oscillatory potentials, and the amplitude of the b-wave of the rabbit electroretinogram were affected; both dopamine and dihydroxyphenylalanine (DOPA) levels were depressed. Ultrastructural analysis of acute and chronic retinas showed the occurrence of an intranuclear filamentous inclusion in the nucleus of some cells in the inner nuclear and ganglion cell layer of chronic samples. The effects of the neurotoxin NMPTP on the retina suggest that dopaminergic metabolism is impaired, which then affects the components of the electroretinogram attributed to both bipolar and amacrine cells. In addition, the retina may provide a model in which to study the bichemical and pharmacological mechanisms of NMPTP toxicity. PMID- 2988990 TI - Immunodetection of Na,K-ATPase in guinea-pig retinal layers, cornea and lens. AB - Na,K-ATPase was detected in crude microsomes of guinea-pig retinal layers, cornea and lens by immunoblotting using antibodies generated against purified kidney Na,K-ATPase. The antiserum cross-reacted with the alpha catalytic subunit of the enzyme from the eye tissues but not the beta glycoprotein subunit, suggesting a high degree of tissue-to-tissue variability in the beta subunit. The apparent molecular weight of the alpha subunit in retinal layers, cornea and lens was 95 Kd which is similar to alpha subunit in kidney. In the retinal layers and cornea, the antiserum detected proteolytic fragments of the enzyme. This method can be used to detect aggregates or proteolytic fragments of Na,K-ATPase in disease states of eye tissues in which electrolyte imbalances have been implicated, such as in experimental and senile cataracts. PMID- 2988991 TI - Suppression of amygdala-kindled seizure in cats by enhanced GABAergic transmission in the substantia innominata. AB - The chronological effect of intracerebral injection of GABAergic drugs, either muscimol or gabaculine, into the substantia innominata, was examined in amygdala kindled cats. Results obtained indicate: that the substantia innominata may play a significant role in ictal linkage between a nonmotor system such as the amygdala and the motor mechanism responsible for amygdala-kindled convulsion, and that the GABA terminals in the substantia innominata exert a suppressive action toward convulsive seizure generalization of amygdala origin. PMID- 2988993 TI - Seroepizootiology of the herpesvirus Papio (HVP) infection in healthy baboons (Papio hamadryas) of high- and low-lymphoma risk populations. AB - Seroepizootiology of Herpesvirus Papio (HVP) infection was studied in three groups of healthy hamadryas baboons (Papio hamadryas): the main Sukhumi (high lymphoma) stock, forest Sukhumi (lymphoma-free) stock and newly imported wild animals. The prevalence to HVP infection, as judged by anti-VCA-HVP positivity, was approximately the same in both Sukhumi stocks (86% and 90% respectively) and it was significantly lower in the pooled group of newly imported baboons. It is interesting that prevalence of HVP infection in the different independent groups varied markedly (35-79%). Geometric mean titers of positives in all groups were approximately the same. The prevalence of HVP infection was age-dependent. It increased during the first years of life reaching the maximum (about 100%) at the age of 5 years being stable up to the age of 18 years and "decreased" at very old ages (over 18 years). The prevalence of HVP infection in newly imported baboons increased with age up to 71% in a group of the "oldest" monkeys and did not plateau. No significant sex differences in anti-HVP titers were found. Anti-EA HVP-positive (with one exception) and anti-HUPNA-positive animals were found only in the main Sukhumi stock. Thus, "serologic activity" against HVP infection was the highest in the ligh-lymphoma stock. PMID- 2988992 TI - Chemical synaptic transmission is not necessary for epileptic seizures to persist in the baboon Papio papio. AB - The spread and persistence of epileptic seizures have generally been attributed to chemical synaptic interactions. Using ion-sensitive microelectrodes, we showed that in the allylglycine-treated photosensitive baboon, prolonged light-induced generalized seizures were accompanied by abnormally large decreases in the concentration of extracellular calcium ions, reaching values at which chemical synaptic transmission was certainly very reduced or blocked. This feature was observed in all cortical layers. Measurements of the concentration of extracellular potassium ions in the course of such light-induced seizures indicated that the observed low values of the concentration of calcium ions could not be ascribed to the occurrence of spreading depressions. These findings showed that nonsynaptic mechanisms play a prominent role in the persistence of epileptic seizures. PMID- 2988994 TI - Skeletal muscle metabolism in mice bearing adenocarcinoma. I. Histochemical alterations in glycogenolytic, glycolytic, lipolytic and oxidative metabolism. AB - Metabolism of triceps, pectoralis (in the vicinity of tumor) and gastrocnemius (away from the tumor) muscles in Swiss albino mice bearing adenocarcinoma has been studied histochemically with regard to content of glycogen, lipids, phosphorylase, aldolase, lipase, succinate dehydrogenase and cytochrome oxidase in the constituent fibres. At 9-10 weeks after transplantation of adenocarcinoma, a negligible glycogen content and decreased phosphorylase and aldolase activities are observed in the white, intermediate and red fibre types in the three muscles. Hypertrophy of fibres and occurrence of targetoid fibres is distinct in the muscles of tumor-bearing mice. The red fibres demonstrate a general loss of lipids, lipase, succinate dehydrogenase and cytochrome oxidase whereas the hypertrophied fibres reveal intense localization of these parameters in their central zones. The results indicate that a decline in glycogenolysis, glycolysis, lipolysis and oxidative metabolism in the various fibre types may contribute to the muscle weakness and muscle wasting in the adenocarcinoma-bearing mice. PMID- 2988995 TI - The influence of metabolic liver defects on diethylnitrosamine (NDEA) carcinogenesis in Gunn rats. AB - Diethylnitrosamine (NDEA) was administered subcutaneously to non-icteric and icteric Gunn rats. A single dose led predominantly to liver cell tumours. The average survival times and tumour latencies were not dose dependent, but a slightly raised tumour multiplicity was observed in icteric animals. The minimal effective dose was apparently less than 0.05 LD50 and more than 0.025 LD50. Chronic treatment in Gunn rats revealed dose response relationships for body weight development, average survival times and tumour latencies. The most common tumours found in the liver of both non-icteric and icteric animals were hepatocellular neoplasms followed by cholangiocellular tumours and haemangio endotheliomas. Neoplastic growth was also found in the respiratory tract and upper digestive tract. The maximal effective carcinogenic dose for non-icteric and icteric rats was 0.1 LD50. The carcinogenic effect of NDEA in Gunn rats was only marginally influenced by the changes in the liver associated with icterus. PMID- 2988996 TI - Nature of the FeO2 bonding in myoglobin: an overview from physical to clinical biochemistry. AB - The iron(II)-dioxygen bond in myoglobin and hemoglobin is a subject of wide interest. Studies range from examinations of physical-chemical properties dependent on electronic structure, to investigations of stability as a function of oxygen supply. Stability properties are of particular importance in vivo, since the oxygenated form is known to be oxidized easily to the ferric form, which cannot be oxygenated and is therefore physiologically inactive. Kinetic and thermodynamic studies of the stability of native oxymyoglobin have revealed a new feature in FeO2 bonding. In vivo, the iron center is always subject to a nucleophilic attack of the water molecule or hydroxyl ion, which can enter the heme pocket from the surrounding solvent, and thereby irreversibly displace the bound dioxygen from MbO2 in the form of O2- so that the iron is converted to the ferric form. A free energy diagram for the potential reactions of FeO2 visualizes myoglobin as a molecular structure that can provide in solution the delicate balance of kinetic and thermodynamic factors necessary to stabilize reversible oxygenation, as opposed to irreversible autoxidation to metmyoglobin. PMID- 2988997 TI - Differential sensitivity to tributyltin of cytochrome-containing and cytochrome deficient cells of Escherichia coli SASX76. AB - The effect of tributyltin (TBT) chloride on the growth of cytochrome-deficient and cytochrome-containing cells of Escherichia coli SASX76 was examined. The former cells were found to be at least 20 times more sensitive to TBT. It is proposed that the differential sensitivity of these two cell types to the biocide, TBT, may be due to a different mode of energy generation by cytochrome deficient and cytochrome-sufficient cells. In addition to the energy state, the pH change caused by the presence and absence of cytochromes which occurred during growth also resulted in a differential sensitivity of these cells. PMID- 2988999 TI - Regional distribution of thiamin pyrophosphokinase in rat brain. AB - The highest specific activity of thiamin pyrophosphokinase was found in the cerebellum, and lower activity in cerebral cortex and midbrain. The regional difference in the enzyme activity was similar to that in thiamin content and the influx rate in rat brain, suggesting that the enzyme is involved in the thiamin transport. PMID- 2988998 TI - Neuromuscular transmission in arterioles. PMID- 2989001 TI - Synthesis and release of plasminogen activator by lymphoid cell lines of malignant origin and its effect on lymphocyte cytotoxicity. AB - Lymphoid B, T. Null and plasma cell lines of malignant origin were evaluated for their ability to produce plasminogen activator (PA) in the 125I-fibrinolysis assay. These cell lines produced PA at varying extent. PA was detected in cell free conditioned medium as well as in cell lysates. The fibrinolysis due to PA was dependent on the presence of plasminogen in the medium. PA purified by affinity chromatography not only abrogated cytotoxicity of peripheral blood lymphocytes significantly, but also the lysis of autologous EBV-transformed cells by in vitro generated cytotoxic lymphocytes. These results indicate that PA produced by malignant cells could inhibit host cellular immunity, thus providing the tumor cells with an escape mechanism from host defences. PMID- 2989000 TI - Inhibition of virus multiplication and alteration of cyclic AMP level in cell cultures by flavonoids. AB - The inhibitory effect of four flavonoid compounds on virus multiplication and their influence on the intracellular cyclic AMP (cAMP) level were studied in cell cultures. Quercetin and quercitrin reduced the yields of Human (alpha) herpesvirus 1 (HSV-1) and Suid (alpha) herpesvirus 1 (pseudorabies virus), but hesperidin and rutin had no effect. Further, quercetin and quercitrin elevated the intracellular level of cAMP, whereas hesperidin and rutin did not alter the cAMP level. Both antiviral activity and cAMP-enhancing effect were dependent on the concentrations of the flavonoids, and these effects turned out to be parallel. This study suggests that a relation exists between the antiviral effect and the cAMP-enhancing activity of flavonoids. PMID- 2989002 TI - Enantioselectivity of the binding of (S)- and (R)-7-chloro-1,3-dihydro-3-methyl-5 phenyl-2H-1,4-benzodiazepines to human serum albumin. AB - By combining the gel filtration and circular dichroism (CD) methods in studying the binding of chiral (S)/(R)-(I) to human serum albumin (HSA) the following results were obtained: HSA affinity for (S)-(I) is about 17 times higher than for (R)-(I); there exist two independent and nonequivalent binding sites for (S)-(I), and one site of lower affinity for (R)-(I); at equimolar concentrations of (I) and HSA, (S)-enantiomer is bound up to 45%, but (R)-enantiomer binds up to 22% only; differential CD-spectra at various concentrations, in the presence of HSA at 1.45 X 10(-5) M concentration, reveals distortions of the chromophoric system i.e. of the conformation of (S)-(I). This effect, and the low affinity of both enantiomers for HSA, allows only a qualitative interpretation of CD-data. PMID- 2989003 TI - Phosphate-binding sequences in nucleotide-binding proteins. AB - In the three-dimensional model of adenylate kinase, the phosphate-binding site for AMP and ATP has been identified [Pai, E.F. et al. (1977) J. Mol. Biol. 114, 37--45]. In this region one can distinguish a sequence glycine XXXX glycinelysine. The same sequence is found in many other mononucleotide-binding proteins including elongation factors and oncogenic P21 proteins. Dinucleotide binding proteins display a pyrophosphate-binding unit with a glycine pattern different from that of mononucleotide-binding proteins. It has been found that P21 ras protein possesses a strand motif typical for (pyro)phosphate binding of a mononucleotide. A single mutation at position 12 can confer oncogenic activity on the protein. Based on the assumption that amino acid residues which are critical for function are preferentially conserved, we predict from the sequence that glycine residue 15 rather than residue 12 is important for (pyro)phosphate binding. PMID- 2989004 TI - Isolation and characterization of cDNA clones for human liver fibronectin. AB - Cellular and plasma fibronectin dimers are constituted by similar but not identical polypeptides. Their differences are the consequence of internal primary sequence variability due to alternative splicing in at least 2 regions (ED and IIICS) of the pre mRNAs [1-8]. A detailed analysis of human liver fibronectin mRNA in these regions was carried out by isolating cDNA clones and determining their nucleotide sequence. A novel type of IIICS segment (coding for 64 amino acids) was present in the two cDNA clones studied and, as expected from previous S1 mapping studies [6], the ED segment was absent in both. PMID- 2989005 TI - Expression of Semliki Forest virus capsid protein from SV40 recombinant virus. AB - Here, the proteolytic processing of the Semliki Forest virus (SFV) capsid protein was studied in the absence of other viral functions. Two different fragments of the SFV messenger cDNA, coding for capsid protein and 174 and 38 extra amino acids from the envelope proteins, respectively, were cloned in the late region of the SV40 viral DNA. Cells infected with the SV40 recombinant virus stocks were analyzed for the production of SFV capsid mRNA and polypeptide. Immunofluorescence staining of the infected cells indicated that the produced SFV capsid protein accumulated mainly in the nucleus. Polyacrylamide gel electrophoresis of the immunoprecipitated SFV capsid proteins showed that both recombinants yielded a labelled band equivalent in size to the SFV capsid protein. Thus the proteolytic processing takes place even under conditions where the capsid protein is the only virus-specified protein synthesized. PMID- 2989006 TI - Cyclic AMP independence of Escherichia coli protein phosphorylation. AB - The effect of cyclic AMP on protein phosphorylation was analyzed comparatively in two strains of E.coli differing in their capacity to synthesize this nucleotide, one of them lacking the adenylate cyclase activity. The results obtained from both in vivo and in vitro experiments concurred in showing that the bacterial protein kinase activity is cAMP-independent. PMID- 2989007 TI - Ubiquinone-5 is reduced by superoxide in the aerobic state by NADPH oxidase of guinea pig macrophages. AB - The reduction of ubiquinone-5 (Q1) by the phagocytosis-specific NADPH oxidase of guinea pig macrophages was not inhibited by superoxide dismutase (SOD) at concentrations usually used for O2- assay but was inhibited at about 100-times higher concentrations. Titration of the reaction with SOD and a comparison with that of xanthine oxidase showed that the inhibition was not due to the semiquinone oxidation accelerated by a removal of O2- but due to the accelerated dismutation of O2- which otherwise reduces the quinone. Molecular oxygens are therefore preferential electron acceptors in the NADPH oxidase even in the presence of Q1. PMID- 2989008 TI - Rapid purification of calcium-activated protease by calcium-dependent hydrophobic interaction chromatography. AB - Both low Ca2+- and high Ca2+-requiring forms of Ca2+-activated protease (calpains I and II) were found to bind to phenyl-Sepharose in a calcium-dependent manner, suggesting that both enzymes expose a hydrophobic surface region in the presence of Ca2+. Inclusion of leupeptin in column buffers prevented the loss of activity during hydrophobic-interaction and substrate-affinity chromatography. Under these conditions calpain II (high calcium-requiring form) was rapidly purified from bovine brain and rabbit skeletal muscle using successive phenyl-Sepharose and casein-Sepharose columns. PMID- 2989009 TI - Phorbol ester inhibits myoblast fusion and activates beta-adrenergic receptor coupled adenylate cyclase. AB - Primary cultures of myoblasts, derived from embryonic chick pectoral muscle, were treated with phorbol ester (TPA) for 8-96 h. TPA treatment blocked the fusion of myoblasts along with the expression of the MM form of creatine kinase. Interestingly, TPA treatment markedly increased the activity of beta-adrenergic receptor coupled adenylate cyclase (AC) activity. The study suggests that TPA treatment augments the functional interaction between a coupling Ns protein and catalytic unit of AC. The likely significance of these results is briefly presented. PMID- 2989010 TI - Induction of early alveolar injury by inhaled asbestos and silica. AB - Inhaled asbestos fibers and silica crystals are known to cause interstitial fibrotic lung disease in animals and humans. The initial cellular events and biochemical mechanisms that lead to development of disease are poorly understood. In ongoing studies reviewed here it has been shown that inhaled particulates small enough to pass through the conducting airways are deposited initially at the bifurcations of alveolar ducts. Within hours after brief exposure, alveolar epithelial cells phagocytose inhaled particles that subsequently are translocated to interstitial matrix and fibroblasts. Within 48 h after exposure, inhaled asbestos on alveolar surfaces activates a complement-dependent chemotactic factor for macrophages that accumulate at duct bifurcations. Epithelial cells, macrophages, fibroblasts, and the interstitial matrix are significantly altered by brief (1- 5-h) exposure to chrysotile asbestos. The basic mechanisms that mediate these initial events remain to be defined. PMID- 2989011 TI - Epithelial properties of brain capillary endothelium. AB - The specialized endothelial cells (ECs) in brain capillaries provide a blood brain barrier to some solutes while facilitating transcapillary exchange of other solutes. In addition, brain capillaries may contribute to the secretion of spinal fluid, a process that is typically mediated by epithelial cells. This proposal is supported by the many epithelial properties of brain capillary ECs including the presence of 1) continuous tight junctions, 2) low transcellular permeability, 3) transcellular concentration gradients, 4) a transcellular potential difference, 5) a high transcellular resistance, and 6) an asymmetrical distribution of transport systems between the luminal and antiluminal plasma membranes. Thus, the brain capillary contains ECs that are structurally and perhaps functionally related to an epithelial cell. These unique features of brain ECs undoubtedly play an important role in regulating the formation and composition of the brain's interstitial fluid. PMID- 2989012 TI - [Interrelation of 5'-nucleotidase separated from thymocytes by heating and receptors responsible for interaction with autologous erythrocytes]. PMID- 2989013 TI - [Clinical and virological studies of eczema herpeticum]. PMID- 2989014 TI - Specific cell-mediated immunity in patients with epidermodysplasia verruciformis and plane warts. AB - The characterization of different types of human papilloma viruses made it possible to study the specific immune responses to purified viral antigens in patients with warts. The specific cell-mediated immunity was investigated by means of a leukocyte migration inhibition factor test in 9 patients with epidermodysplasia verruciformis and 4 patients with regressing plane warts. The results show a significant increase in specific cell-mediated reactivity concomitant with the regression of the warts. PMID- 2989015 TI - Lack of beta-adrenergic role for catecholamines in the development of hyperglycemia and ketonaemia following acute insulin withdrawal in type I diabetic patients. AB - In order to evaluate the role of beta-receptor mediated effects of catecholamines in the metabolic deterioration following insulin withdrawal in insulin-dependent diabetic patients we have measured in 5 patients metabolic substrate and hormone concentrations during a 6 hours arrest of insulin infusion, without or with a simultaneous infusion of propranolol. During insulin deprivation plasma epinephrine and norepinephrine increased slightly (from 107 +/- 10 ng/L to 173 +/ 6 ng/L and from 307 +/- 37 ng/L to 518 +/- 77/ng/L respectively (p less than 0.05), cortisol decreased physiologically, but growth hormone and glucagon were not significantly modified. Free insulin decreased progressively from 12.2 +/- 2.5 mU/L to 5.4 +/- 1.1 mU/L (p less than 0.01). Blood glucose and ketone bodies rose sharply before any significant change in catecholamine levels. Plasma free fatty acids and blood glycerol increased progressively and their rise appeared somewhat temporally related to the variations of catecholamine levels. The addition of propranolol to insulin deprivation did not modify the changes in hormone concentrations in spite of a slightly greater rise of epinephrine (from 78 +/- 4 ng/L to 179 +/- 7 ng/L, p less than 0.05) and norepinephrine (from 395 +/- 80 ng/L to 679 +/- 153 ng/L, p less than 0.05). The rises of glucose and ketone bodies were unaffected whereas the increases of free fatty acids and glycerol were slightly blunted. In conclusion, we have no evidence for a beta adrenergic mediated role for catecholamines in the development of hyperglycaemia and ketonaemia in non-stressed insulin deprived diabetic patients, and only small evidence for a permissive effect on lipolysis. PMID- 2989017 TI - Alterations in the activities of subcellular fractions marker enzymes in rat liver and brain by hydrocortisone and corticosterone treatment. AB - The effect of subcutaneous injection of hydrocortisone and corticosterone on the activity values of some subcellular fractions marker enzymes from rat liver and brain was investigated and compared with controls (without treatment with hormones). The following enzymes were studied (subcellular fraction are shown between parentheses): N-acetyl-beta-D-glucosaminidase and beta-glucuronidase (lysosomes); succinate dehydrogenase = SDH (mitochondria); glucose-6-phosphatase (endoplasmic reticulum); 5'-nucleotidase and Na+-K+-Mg2+ ATPase (plasma membrane). The specific activity of lysosomal enzymes from liver showed no change when rats were injected either with hydrocortisone or corticosterone. The same enzymes from brain showed significant increases in their activities with both hydrocortisone or corticosterone except beta-glucuronidase; this enzyme gave activity values remaining between the control levels, after treatment with corticosterone. The activity of mitochondrial SDH was increased after corticosterone injection either in liver or brain. After hydrocortisone injection, its activity rises significantly in brain (72%), but it falls in liver compared to the control values. Glucose-6-phosphatase behaves similarly in brain or liver fractions; its activity increases always after corticosterone treatment and decreases by hydrocortisone. The plasma membrane marker enzymes did not change practically in brain fractions, excepted Na+-K+-Mg2+ ATPase which tends to rise its activity after hydrocortisone injection. In liver fractions, both 5' nucleotidase and Na+-K+-Mg2+ ATPase activities increase either by corticosterone or hydrocortisone treatment, except 5'-nucleotidase which specific activity decreases in liver after hydrocortisone treatment. PMID- 2989016 TI - Particles and macrophages in murine Peyer's patches. AB - Mice were given 1% suspensions of 5 insoluble particles (chrysotile asbestos, quartz, carmine, carbon, and iron oxide) in drinking water for 3 months. The particles were subsequently sought in intestinal Peyer's patches by light microscopy. Carbon and iron oxide particles were visible in Peyer's patch macrophages, particularly in the subepithelial region, but the other particles could not be detected. The findings suggest that particle surface properties as well as particle size govern accumulation in Peyer's patches. The cytochemistry of subepithelial, mid-dome, tingible-body, and serosal macrophages of control mice indicated diversity of macrophages within the patch. Macrophages of asbestos fed mice contained more lysosomes than macrophages of controls. Macrophage abundance in the dome apex was not significantly altered by asbestos ingestion. The other particles did not produce detectable alterations in macrophage morphology. PMID- 2989018 TI - Study of carbohydrate metabolism in glycogen storing cell lines derived from cultured rat hepatocytes. AB - Some aspects of carbohydrate metabolism were investigated in three non-malignant, glycogen storing, cell lines derived from a primary culture of rat hepatocytes, and in the Morris hepatoma 3924 cells. The three cell lines show biochemical alterations which are, to a large extent, similar to those found in the hepatoma cells: increased activity of glycolytic enzymes and decreased activity of gluconeogenetic enzymes. An increase of glucose-6-phosphate dehydrogenase activity is also found. The three cell lines, as the Morris hepatoma cells, actively convert glucose into lactate under the in vitro conditions of culture. Fructose is not taken up as quickly as glucose and galactose is not metabolized. As compared with normal hepatocytes, the three cell lines have altered metabolism and growth behaviour. They largely resemble the preneoplastic cells appearing in rat liver at the early stages of experimental carcinogenesis. PMID- 2989019 TI - Acid phosphatase and phosphoamino acid phosphatases in murine erythroleukaemic cells. AB - The activities of acid and alkaline phosphatases and phosphotyrosine, phosphoserine and phosphothreonine phosphatases were measured in Friend murine erythroleukaemic (MEL) cells. The effects of treating the cells with dimethyl sulphoxide (DMSO), an inducer of differentiation, were examined. In untreated cells alkaline phosphatase activity was undetectable, though there were significant amounts of acid phosphatase (76 +/- 15 mU/mg protein) and phosphotyrosine phosphatase (16 +/- 0.9mU/mg protein); phosphoserine and phosphothreonine phosphatase activities (9 +/- 0.4 and 7 +/- 0.6mU/mg protein, respectively) were lower than for phosphotyrosine phosphatase. Addition of 1 or 2% DMSO to the culture medium resulted in the expected cell death within 2 weeks. With 0.5% DMSO, cells remained viable for at least 8 weeks, but while some appeared to have smaller nuclei and retained their rounded appearance, others became fibroblastic within several days and adhered to the culture vessel. The treated cells which had kept their morphology showed no difference in acid phosphatase activities as compared with untreated controls; phosphotyrosine phosphatase was lower (9 +/- 0.8mU/mg protein) and phosphoserine and phosphothreonine phophatases higher (11 +/- 0.5 and 10 +/- 0.4mU/mg protein, respectively) than in the controls. The Km values for p-nitrophenyl phosphate were similar in untreated and treated cells (0.069 and 0.068mM, respectively); for phosphotyrosine the Km value was lower in the treated cells (0.97mM) than in the controls (1.9mM). PMID- 2989021 TI - Formation of HCN by human phagocytosing neutrophils--1. Chlorination of Staphylococcus epidermidis as a source of HCN. AB - Phagocytosis of Staphylococcus epidermidis by human neutrophils is accompanied by HCN liberation. The amount of HCN evolved is significantly higher when the bacteria are damaged by penicillin. One of the substrates yielding HCN during phagocytosis are N-dichloroglycyl residues of bacterial peptidoglycan formed by the chlorinating myeloperoxidase-H2O2-Cl- system of neutrophils. HCN deriving from the bacterial structures constitutes 6-12% of total HCN liberated during phagocytosis, which indicates that there are other substrates for HCN production. Chlorination process within phagocytosing neutrophils seems to be essential for formation of both, total HCN and that deriving from bacterial structures. PMID- 2989020 TI - Studies on the diurnal rhythm of mevalonate metabolism by sterol and nonsterol pathways and of mevalonate-activating enzymes. AB - Both in vivo and in vitro incorporation of mevalonic acid into nonsaponifiable lipids by 17-day-old chick liver and kidney did not show diurnal rhythm. Using 14CO2 production from MVA as an index of the shunt pathway not leading to sterols, we have demonstrated for the first time that there is no diurnal rhythm in this pathway. No significant differences were found in the specific activities of mevalonate kinase, mevalonate-5-phosphate kinase and mevalonate-5 pyrophosphate decarboxylase from chick liver and kidney throughout a period of 24 hr, using [1-14C]mevalonate as substrate. The absence of diurnal rhythm in the decarboxylase activity was corroborated by further experiments carried out using [2-14C]mevalonate-5-pyrophosphate as specific substrate of this enzyme. PMID- 2989022 TI - T-lymphoblast-specific nucleoside kinase: characterization and comparison with deoxycytidine kinase. AB - The results of Sephadex G-100 gel filtration and sucrose density gradient centrifugation showed that T-lymphoblast-specific nucleoside kinase (TSK) purified from MOLT 4FT cell extract has a molecular weight of 26,500, while deoxycytidine kinase (dCK) 56,000. The pI value of TSK (pH 8.2) is quite different from that of dCK (4.8). TSK phosphorylated deoxycytidine, deoxyadenosine, deoxyguanosine and arabinocytidine, similar to dCK, but the respective kinetic properties were quite different. In the phosphate donor specificity and metal ion requirement, some differences were observed between TSK and dCK. dTTP was a good phosphate donor for dCK but no effect at all as phosphate donor for TSK. dCK was inhibited at very low concentration of dCTP, but TSK at much higher concentration of dCTP. PMID- 2989024 TI - Glycosidases in normal and regenerating chicken liver, hepatoma Mc-29, Rous sarcoma, in turkey poult liver and hemocytoblastomes, provoked by the leukosis virus strain Mc-31. AB - Four glycosidases (beta-galactosidase, alpha-mannosidase, alpha-fucosidase and beta-N-acetylglucosaminidase) were studied in chicken normal and regenerating liver, in turkey poult liver and in virus induced avian tumors--chicken hepatoma (strain Mc-29), Rous sarcoma (strain Schmidt-Ruppin) and turkey poult hemocytoblastoma nodules (strain Mc-31). The multiple forms of beta-N acetylglucosaminidase were assayed as well. A particular enzyme pattern was found in the tumor lines under investigation. A characteristic property of hepatoma cells was the elevation of beta-galactosidase activity and of the former enzyme and that of beta-N-acetylglucosaminidase for the hemocytoblastoma. In Rous sarcoma the glycosidase activities (except that of alpha-fucosidase) were much lower, compared to the other two solid tumors. All enzyme activities were compared with those in the normal liver of the corresponding avian species, and with the liver of tumor bearing fowls and with regenerating chicken liver. Unlike the rat liver in the avian normal and tumor tissues the percentual ratio between the multiple forms A and B of beta-N-acetylglucosaminidase was found to be 30:70%. PMID- 2989023 TI - Biochemical basis of muscular fatigue associated with repetitious contractions of skeletal muscle. AB - The decline in force generating capabilities of skeletal muscle associated with prolonged, repetitive low force producing contractions does have a biochemical basis. It is our view that an alteration in neuromuscular transmission results in an uncoupling of excitation-contraction via disturbances in Ca2+ imbalance, an uncoupling of energy utilization and production may result, which affect a favourable cellular environment for the initiation of myofilament degradation. The myofilament dissolution may be the last stage in this fatigue process and associated with only extreme conditions of muscle use. PMID- 2989025 TI - Trifluoperazine abolishes the actions of bradykinin on glucose 1,6-bisphosphate levels and on the activities of glucose 1,6-bisphosphatase, phosphofructokinase and phosphoglucomutase. AB - Injection of trifluoperazine abolished the bradykinin-induced decrease in intracellular concentration of glucose 1,6-bisphosphate (Glc-1,6-P2) in rat tibialis anterior muscle and skin. These changes in Glc-1,6-P2 levels may be attributed to the changes in the activity of glucose 1,6-bisphosphatase (the enzyme that degrades Glc-1,6-P2), which was markedly enhanced by bradykinin and reversed by trifluoperazine. Concomitantly to the changes in Glc-1,6-P2, the potent activator of phosphofructokinase and phosphoglucomutase, the activities of these enzymes were reduced by bradykinin and restored by trifluoperazine. These findings suggest that trifluoperazine treatment may have a beneficial effect on the depressed glycolysis induced by bradykinin in tissue damage. PMID- 2989026 TI - Metabolic effects of buflomedil hydrochloride. PMID- 2989027 TI - Modulation of cultured mouse Leydig cells adenylate cyclase by forskolin and hCG. AB - The diterpene, forskolin, stimulated cAMP accumulation about 15-fold over basal levels in purified mouse Leydig cells; however, it remained far less potent than hCG. Simultaneous addition of forskolin and hCG resulted in a striking synergistic stimulation of cAMP production. In contrast, forskolin-enhanced testosterone accumulation was never synergistic with that produced by maximal concentrations of hCG. hCG (3 X 10(-9) M) lowered about 6-fold the ED50 for forskolin-elicited cAMP accumulation and increased the maximal response to forskolin about 16-fold. Conversely, forskolin 10(-6) M) reduced the ED50 for hCG 2-fold but had a much smaller effect (2-3-fold) on maximal response. Moreover, pretreatment with hCG induced only a homologous desensitization of adenylate cyclase, whereas the enzyme became partially resistant to both hCG and forskolin in cells pretreated with forskolin. The homologous hCG-induced desensitization and the partial heterologous one induced by forskolin suggest that more than the catalytic unit of the cyclase is required for the diterpene activation. PMID- 2989028 TI - Evidence for rapid loss of spare hCG receptors in the corpora lutea of the hypophysectomized rat. AB - In pregnant mare serum gonadotropin treated immature rats hypophysectomized on the day of ovulation (day 1) the corpora lutea (CL) persist as normal morphological structures and produce steroids, especially 20 alpha dihydroprogesterone (20 alpha-DHP), for at least 40 days (Taya and Greenwald, 1982), although there is a rapid decline in human chorionic gonadotropin (hCG) binding sites (Kim and Greenwald, 1984). In this study the number of occupied and non-occupied hCG receptors in CL from hypophysectomized rats decreased to 14% and 36%, respectively, compared to intact day 5 pseudopregnant animals, but the binding affinity was unchanged. Decreased concentration of occupied hCG receptors paralleled hormonal levels of serum luteinizing hormone (LH) which were measurable in only 9 out of 20 animals and were near the lower limits of the assay (40 pg/ml). Luteal progesterone (P4) production in response to LH was markedly decreased after hypophysectomy, but the maximal P4 response to LH and 20 alpha-DHP production in response to LH and 8-Br-cAMP were the same in hypophysectomized and intact animals. Although hCG receptor concentration in CL decreased significantly after hypophysectomy, LH-stimulated luteal production of cAMP was almost the same in both groups. These results indicate that LH spare receptors which are uncoupled from cAMP exist in the CL of the intact pseudopregnant rat and that after hypophysectomy they quickly disappear within 4 days. PMID- 2989029 TI - Down-regulation of nuclear T3 receptors by thyroid hormones in the rat anterior pituitary. AB - The concentration of T3 nuclear receptors may be modulated by thyroid hormones. We have examined the effect of thyroid hormone excess and deficiency in the rat on the in vitro binding of T3 to anterior pituitary nuclear receptors. Nuclei prepared from hypothyroid, hyperthyroid and control rats by homogenisation and centrifugation were incubated with [125I]T3 and unlabelled T3 at increasing concentrations. After incubation for 60 min at 37 degrees C, nuclei were separated by filtration and bound radioactivity counted. Scatchard analysis of binding data revealed no significant difference in binding affinity or capacity for T3 between nuclei from hypothyroid and control rats. Results from hyperthyroid and control animals revealed no change in affinity, but a marked reduction in binding capacity was observed, suggesting depletion of receptor number. This finding of down-regulation may represent a protective mechanism against the effects of high circulating thyroid hormone levels on the tissues. PMID- 2989030 TI - Effect of thyroid status on the development of the different molecular forms of Na+,K+-ATPase in rat brain. AB - The effect of thyroid status on the postnatal development of the two molecular forms of Na+,K+-ATPase, distinguished kinetically on the basis of their ouabain sensitivity, was examined in rat brain. Hypothyroidism induced by PTU from day 1 postnatally significantly reduced the Na+,K+-ATPase activity in cerebellum (22-30 days) but not forebrain, whereas hyperthyroidism (T4 treatment from day 1) had no effect. The hypothyroidism-induced reduction in cerebellum was reflected by a 20 45% reduction in the activity of the alpha(+) form of Na+,K+-ATPase (high ouabain affinity) against control brains compared to a 60-70% reduction in the activity of the alpha form (low ouabain affinity). These results show that neonatally induced hypothyroidism leads to a selectively greater impairment of the ontogenesis of the activity of cerebellar alpha form of Na+,K+-ATPase. This may possibly reflect a retarded development of a selective cerebellar cell population containing predominantly the alpha form of the enzyme. PMID- 2989031 TI - Inhibition by estradiol of binding and mitogenic effect of epidermal growth factor in primary cultures of Xenopus hepatocytes. AB - We demonstrate here the presence of two classes of EGF (epidermal growth factor) binding sites in primary cultures of male Xenopus liver parenchymal cells. One of these corresponds to the high-affinity receptor described in other tissues and species, and which exhibits the property of autophosphorylation. The number of EGF receptors decreased sharply in freshly prepared cultures but recovered to maximum levels within 24 h thereafter. Addition of EGF and insulin to the hepatocyte cultures enhanced the rate of DNA synthesis as measured by the incorporation of [3H]thymidine. Estrogen abolished this increase, reducing the incorporation to that seen with hydroxyurea. At the same time, the addition of estradiol reduced the number or activity of EGF receptors in a dose-dependent manner. The latter paralleled the activation of transcription of vitellogenin genes in Xenopus hepatocytes so that a high rate of DNA synthesis is unnecessary for or incompatible with the activation of the steroid hormone-induced vitellogenin genes. PMID- 2989033 TI - Regulation of aromatase activity of rat granulosa cells: induction of synthesis of NADPH-cytochrome P-450 reductase by FSH and dibutyryl cyclic AMP. AB - Aromatase is an enzyme complex that is composed of a specific form of cytochrome P-450 and a flavoprotein, NADPH-cytochrome P-450 reductase. Aromatase activity of granulosa cells is increased markedly by follicle-stimulating hormone (FSH) and by analogs of cyclic AMP. It was the objective of the present study to investigate the effects of FSH and dibutyryl cyclic AMP (Bt2cAMP) on the synthesis of NADPH-cytochrome P-450 reductase in rat granulosa cells maintained in vitro. Granulosa cells were obtained from the ovaries of diethylstilbestrol (DES)-treated immature rats and were incubated in the presence of DES (10(-7) M), DES + FSH (250 ng/ml), or DES + Bt2cAMP (1 mM) for up to 72 h. After 72 h of incubation, aromatase activity of cells incubated with DES alone was 5 pmoles estrogen formed 2 h-1 mg-1 protein and was increased greater than 60-fold in cells incubated with FSH or Bt2cAMP. NADPH-cytochrome P-450 reductase was immunoisolated from [35S]methionine-labeled lysates of granulosa cells incubated for 72 h in the absence or presence of stimulatory factors. The rate of synthesis of reductase was found to be increased about 3-fold in cells incubated with DES + FSH or DES + Bt2cAMP as compared to cells incubated with DES alone. By immunoblot analysis we found that the cellular content of reductase was increased about 2 fold by FSH and Bt2cAMP treatment. Reductase specific activity was 10 nmoles min 1 mg-1 protein in membrane fractions of DES-treated cells and was increased 1.6 fold by FSH treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989032 TI - Desensitization of LLC-PK1 cells by vasopressin results in receptor down regulation. AB - The molecular mechanism of desensitization of antidiuretic hormone receptors is not well understood. Preincubation of LLC-PK1 cells with lysine vasopressin (LVP) (10(-6) M, 5 h) decreased subsequent LVP-stimulated cAMP accumulation in cells by 83% and reduced the Vmax of LVP-stimulated adenylate cyclase by 81%. Such preincubation also reduced by 90% the binding of [3H]LVP to both intact cells and isolated plasma membranes, suggesting a loss of vasopressin receptors. Both the reduction in cAMP response and the apparent loss of receptors showed similar dose and time dependence. Monensin (33 microM) did not alter [3H]LVP binding or stimulation of cAMP by LVP, nor did it prevent desensitization. However, membranes prepared from cells preincubated with LVP in the presence of monensin did not show a decrease in [3H]LVP binding. Forskolin preincubation, at 0.1, 1, 10 and 100 microM, did not alter [3H]LVP binding or accumulation of cellular cAMP by LVP, nor did it induce desensitization to LVP. Cells desensitized with varying LVP concentrations in the presence of 10 microM forskolin displayed the same loss of [3H]LVP binding and LVP responsiveness as observed in the absence of forskolin. LVP-desensitized cells, upon removal from LVP-containing medium, recovered cAMP responsiveness to LVP and specific binding of [3H]LVP at the same rate, achieving control levels after 50 h. Recovery was prevented by cycloheximide (25 micrograms/ml). These findings are consistent with a desensitization process involving LVP-mediated receptor internalization, and a recovery process requiring protein synthesis. PMID- 2989034 TI - The role of calcium in diuretic hormone action on locust Malpighian tubules. AB - Fluid secretion by the Malpighian tubules (MTs) of Locusta is drastically reduced in the absence of extracellular calcium. Verapamil (10(-4) M) inhibits basal secretion, whereas the ionophore A23187 (10(-5) M) elevates the secretory rate. Cyclic guanosine monophosphate (cGMP) stimulates fluid secretion at a concentration of 10(-3) M. A factor extractable in methanol from the storage lobes of the corpora cardiaca stimulates increased guanylate cyclase activity in MTs, resulting in a 10-fold elevation in intracellular cGMP levels. Attempts to separate the factor stimulating guanylate cyclase by high-performance size exclusion chromatography proved unsuccessful. Neither 5-hydroxytryptamine (5-HT) (10(-4) M) nor A23187 (10(-5) M) are able to elevate intracellular cGMP levels. Elevations of both intracellular cAMP and cGMP levels in response to diuretic hormone (DH) are potentiated in the absence of extracellular calcium. Consistent with these elevations are increases in the rates of fluid secretion by tubules deprived of extracellular calcium. It is concluded that, although calcium has an important role in the regulation of fluid secretion, its role in the mechanism of hormone-stimulated secretion may be modulatory rather than regulatory. PMID- 2989035 TI - Prolactin receptor in rat liver: sex difference in estrogenic stimulation and imprinting of the responsiveness to estrogen by neonatal androgen in male rats. AB - Rat hepatic prolactin receptor is regulated by sex steroids. A high level of the receptor was found in female rats but the level was nearly undetectable in males. Gonadectomy reduced the receptor level in females but increased the level in males. Administration of estradiol benzoate (0.05 mumoles/kg on alternate days subcutaneously for 9 days) to adult gonadectomized females increased the receptor level by 473% whereas the same treatment in adult gonadectomized males produced a more modest 276% increase. This sexually dimorphic pattern in the responsiveness to estrogen stimulation in adult rats appeared to be determined neonatally. Neonatal gonadectomy of male rats changed the hepatic response system to a more female pattern in adulthood. Replacement of testosterone (1.45 mumoles at days 1 and 3 after birth) to these neonatally gonadectomized male rats restored the male pattern. Diethylstilbestrol replacement (1.45 mumoles at days 1 and 3 after birth) to the neonatally gonadectomized male rats showed the same effect as neonatally administered testosterone. Scatchard analysis revealed that the observed changes in binding are related to changes in binding capacity but not affinity. Desaturation by 4 M MgCl2 indicated that the amount of endogenously bound hormone was negligible in our membrane preparations. PMID- 2989037 TI - Polyphosphoinositide metabolism in adrenal glomerulosa cells. AB - We examined the effect of angiotensin II, a calcium-mobilizing hormone on polyphosphoinositide metabolism in isolated rat adrenal glomerulosa cells. In cells preloaded with [32P]phosphate or with [3H]inositol, stimulation with angiotensin resulted in an approx. 40% reduction in the radioactivity of triphosphoinositide (PtdIns4,5P2) within 15 s. Only a slight increase in radioactivity was observed in the subsequent 30 min. Changes in labelling of diphosphoinositide (PtdIns4P) showed similar kinetics. Incorporation studies also showed a reduction in the pool size of [32P]PtdIns4P and [32P]PtdIns4,5P2 in response to angiotensin. Production of inositol phosphates in the absence or presence of lithium, a cation-inhibiting myo-inositol-1-phosphatase, was examined in cells preloaded with [3H]inositol. The results indicate that the production rate of inositol tris- and bisphosphate shows a manifold increase in the first seconds of stimulation and remains enhanced for at least several minutes. The present data suggest that the rate of resynthesis of polyphosphoinositides also increases shortly after the activation of PtdIns4,5P2 phosphodiesterase. Corticotropin, a hormone acting via cyclic AMP, neither affected polyphosphoinositide metabolism nor modified the action of angiotensin II. PMID- 2989036 TI - Control of secretion and the function of C19-delta 5-steroids of the human adrenal gland. PMID- 2989038 TI - Inhibition of cyclic AMP formation by iodide in suspension cultures of porcine thyroid follicle cells. AB - In the present study porcine thyroid cells in suspension cultures were employed to investigate the suppressive effect of iodide on adenylate cyclase under basal conditions and following incubation with TSH, PGE1, cholera toxin and forskolin. Within 30 min of incubation with iodide (half-maximal effect 10(-5) M), inhibition was established and remained unchanged up to 40 h of culture. The inhibitory action was abolished by methimazole. TSH, PGE1, cholera toxin and forskolin stimulated cAMP accumulation 10-, 3-, 24- and 22-fold, respectively. Iodide pretreatment reduced basal cAMP levels and also made the cells less sensitive to stimulation by the various agents. High concentrations of TSH or PGE1 could not overcome the suppressive influence of iodide, whereas with high concentrations of cholera toxin and forskolin the reduction in cAMP levels in iodide-treated cultures was less pronounced. Membranes isolated from iodide treated cultures produced significantly lower amounts of cAMP compared to control membranes. Furthermore, iodide did not inhibit basal or forskolin-stimulated cAMP production in human fibroblasts. The results demonstrate that iodide via an iodination-dependent mechanism influences cAMP generation in thyroid cells. It is suggested that the inhibitory activity, which has a long half-life, involves stable modification of the membrane-localized catalytic unit of adenylate cyclase such that its activation by the regulatory unit is rendered less efficient. PMID- 2989039 TI - Increased gonadotrophin releasing hormone receptors on pituitary gonadotrophs: effect on subsequent LH secretion. AB - GnRH, high potassium concentrations, and cAMP derivatives have been previously shown to increase GnRH receptor levels (GnRH-R) in cultured rat pituitary cells. However, the effect of these changes in receptor number on subsequent stimulated LH release has not been investigated. In this study pretreatment of pituitary cells with either 1 nM GnRH, 58 mM KCl, or 1 mM dibutyryl cAMP (dbcAMP) resulted in a 70-100% increase in GnRH-R 7-10 h later. Subsequent LH responses to GnRH in those cells pretreated with GnRH and KCl were markedly reduced and the dose response characteristics altered such that the curves were non-sigmoidal. When corrected for depletion of cellular LH during the pretreatment period these GnRH response curves were similar to control, implying that hormone depletion was the explanation for apparent desensitisation. By contrast, dbcAMP and low-dose calcium ionophore (0.1 microM A23187) pretreatment, which did not deplete cellular LH, neither enhanced nor decreased subsequent sensitivity to GnRH. Thus, 4 agents which all, under these conditions, increased GnRH receptors did not sensitise gonadotrophs to GnRH. By contrast, pretreatment with 10(-9) and 10(-8) M GnRH for either 12 or 16 h rendered cells completely or partially refractory to further GnRH stimulation, despite an increase in GnRH receptors. This desensitisation could not be explained by cellular LH depletion, and was specific to the homologous ligand since dose-responses to the Ca2+ ionophore A23187 and KCl were normal when corrected for LH depletion. Non-receptor-mediated depletion of cellular LH during A23187 pretreatment (10 microM for 10 h) did not alter subsequent GnRH dose-responses, after correction for LH content. These data indicate that, under these in vitro conditions, the increased GnRH receptors are not functionally linked to the secretory apparatus of the gonadotroph. Furthermore, homologous ligand-induced desensitisation is both time- and concentration-dependent and is mediated largely by post-receptor cellular events independent of cellular LH content. Therefore, post-receptor cellular processes may be more important than changes in GnRH receptors in regulating gonadotrophin secretion. It is suggested that an increase in GnRH receptors may represent a cellular response to generalised gonadotroph activation by a variety of agents, and does not necessarily signify enhanced responsiveness to GnRH. PMID- 2989040 TI - Receptor-mediated degradation of human growth hormone in rat adipocytes and cultured human lymphocytes (IM-9). AB - In cultured human lymphocytes (IM-9) and in isolated rat adipocytes human growth hormone is substrate for a receptor-mediated degradation. When the cells are incubated with monoiodinated human growth hormone half of the radioactivity dissociating from the cells is in the form of [125I]monoiodotyrosine. Since IM-9 lymphocytes have no receptor-mediated degradation of insulin, obviously insulin and human growth hormone follow different pathways in this cell type. In the rat adipocyte colchicine and monodansylcadaverine caused quantitatively different uptake and degradation of these 2 ligands suggesting that also in this cell type the pathways are functionally different. The application of different inhibitors suggests that the receptor-mediated degradation of growth hormone in these 2 cell types takes place in an acidified compartment by an energy-requiring process and involving thiol groups. PMID- 2989042 TI - Production of PDGF-like growth factors by embryonal carcinoma cells and binding of PDGF to their endoderm-like differentiated cells. AB - In this report, we demonstrate that F9 and PC-13 embryonal carcinoma (EC) cells do not bind significant amounts of platelet-derived growth factor (PDGF), whereas the endoderm-like differentiated cells derived from EC cells do. The F9 differentiated cells exhibit approximately 8300 receptors per cell, with an apparent dissociation constant of 30 pM. Two endoderm-like cell lines, PSA-5E and PYS-2, also bind PDGF and exhibit approximately 4800 and 23,500 receptors per cell, respectively. The lack of PDGF binding by the parental EC cells is consistent with their release of a factor(s) that is closely related to PDGF. This factor(s) competes with PDGF for binding to membrane receptors and is recognized by antibodies raised against PDGF. However, this factor(s) does not appear to be antigenically identical to PDGF. We also show that production of this PDGF-like factor(s) is reduced more than 90% when F9 EC cells differentiate into cells that bind PDGF. Thus, our findings indicate that EC cells release a factor(s) that should be capable of binding to their differentiated cells. This raises the possibility that PDGF, or a closely related factor, can influence cell proliferation and/or cell behavior of early embryonic cells. PMID- 2989041 TI - Interaction of mouse prolactin with mouse hepatic receptors. AB - Lactogenic receptors are usually studied in heterologous systems where prolactin is derived from one species and receptors prepared from another. In such systems the foreign prolactin could be seen as a growth hormone by the host tissue. We have therefore developed a homologous radioreceptor assay using secreted mouse prolactin (smPRL) and mouse hepatic receptors. In this system, monovalent anions augment the smPRL-receptor interaction in the order F- greater than Cl- greater than Br- greater than I-. Divalent cations (Mg2+, Ca2+, Sr2+), phosphate and acetate also increase smPRL binding. Temperature and pH optima are at 8 degrees C and pH 8.3, respectively. Under optimum conditions, the percent total, specific and nonspecific binding are 55%, 45% and 10%, respectively. At infinite receptor concentration the maximum specific bindability of labeled smPRL is 50%. The effects of ions on binding of smPRL to the receptor show that hydrophobic forces participate in smPRL-receptor coupling. The biphasic dissociation kinetics show initial and final rate constants of 1.56 X 10(-4)/s and 7.62 X 10(-6)/s, respectively. The lactogenic receptor does not bind mouse growth hormone; however, it binds both mouse placental lactogen (mPL) and smPRL with equilibrium association constants of 3.90 X 10(8) M-1 and 2.25 X 10(8) M-1, respectively, suggesting that smPRL and mPL share biological roles by acting through the same receptor. PMID- 2989043 TI - A characterization of the preaggregative period of Dictyostelium discoideum. AB - The preaggregative period of Dictyostelium discoideum has been characterized by measuring the reduction in time for the onset of aggregation under conditions which hinder close cell-cell associations, inhibit protein synthesis, and/or include continuous high concentrations or pulsed low concentrations of exogenous cAMP. The results demonstrate that: the preaggregative period (normally 7 hr for cells from log phase cultures) can be dissected into two distinct components: an initial component which includes the first 4.5 hr, and a second component which includes the last 2.5 hr; the first component will progress at normal rate in the continuous absence of close cell-cell associations (as single amoebae in suspension) or in the continuous absence of de novo protein synthesis; the second component will not progress in the continuous absence of close cell-cell associations or de novo protein synthesis; high concentrations of cAMP continuously present in suspension cultures do not affect progress through the first component, nor do they support progress through the second component; however, if cells are allowed to form close cell-cell associations during progress through the first component, high concentrations of cAMP will support progress through the second component in the absence of close cell-cell associations; these associations, which render cells sensitive to cAMP, will occur in the absence of de novo protein synthesis and before the acquisition of contact sites A; these associations may be completely bypassed if suspended cells are continuously pulsed with low concentrations of cAMP; in this case, pulses of cAMP will support progress through the final component in continuous suspension cultures; and the acquisition of contact sites A will not occur in the absence of progress through the second component; in contrast, the acquisition of cAMP binding sites on the cell's surface will occur. These results are considered in terms of the complexity and regulation of the preaggregative period of Dictyostelium. PMID- 2989044 TI - Regulation of protein synthesis during the preaggregative period of Dictyostelium discoideum development: involvement of close cell associations and cAMP. AB - The preaggregative period of Dictyostelium discoideum is composed of two rate limiting components which exhibit dramatic differences in either their dependency upon, or sensitivity to, close cell-cell associations, inhibitors of protein synthesis, temperature, and pH. The first component comprises the initial 4.5 hr and the second component the last 2.5 hr of the preaggregative period. By pulse labeling cells with [35S]methionine, separating polypeptides by 2D-PAGE, and semiquantitatively comparing the rates of synthesis of 778 individual polypeptides by fluorography, the following results were obtained: a detailed program of protein synthesis accompanies the preaggregative (0-7 hr) and aggregative (7-10.5 hr) periods of development; this includes significant decreases in the rate of synthesis of 93 polypeptides synthesized during vegetative growth and significant increases in the rate of synthesis of 74 polypeptides either undetectable or synthesized at relatively low rates during vegetative growth; 35 polypeptides are transiently synthesized at different times during the preaggregative and aggregative periods; two peaks of activity are clearly defined for both increases and decreases; these peaks correlate temporally with the first and second rate-limiting components of the preaggregative period; the majority of changes (74%) which occur during the first rate-limiting component will occur in the absence of close cell-cell associations, but the majority (66%) which normally occur during the second rate limiting component do not occur in the absence of close cell-cell associations; a high concentration of cAMP in the medium of continuous suspension cultures does not stimulate most of the changes which are dependent upon close cell-cell associations; even though cAMP stimulates progress through the second rate limiting component in suspension cultures first allowed to associate for 4.5 hr ("competent" cells) prior to disaggregation it still does not stimulate most of the changes which are dependent upon close cell-cell associations; and synthesis of only 3 out of 778 polypeptides appears to be stimulated by addition of exogenous cAMP, and only in resuspended cultures of "competent" cells. The prominent role of close cell-cell association and the surprisingly minor effect of cAMP in the regulation of the program of protein synthesis accompanying the preaggregative and aggregative periods of Dictyostelium are discussed, especially as they relate to the effect of cAMP on protein synthesis in suspended cultures of postaggregative cells. PMID- 2989045 TI - Release of plasminogen activator and a calcium-dependent metalloprotease from cultured sympathetic and sensory neurons. AB - Cultures of neurons from neonatal rat superior cervical, dorsal root, and trigeminal ganglia were grown in the absence of nonneuronal cells in serum-free defined medium. Proteins metabolically labeled with radioactive amino acids and spontaneously released into the culture medium were studied using two-dimensional gel electrophoresis and photofluorography. All three populations of neurons released 12-15 major proteins into the culture medium. Four proteins were released selectively by sympathetic neurons and two proteins were consistently released by both populations of sensory neurons but not by sympathetic neurons. Enzymatic activities are associated with at least two of the released proteins. One is a calcium-dependent metalloprotease, and the other a plasminogen activator. The calcium-dependent metalloprotease has a MW of 62 kDa, requires millimolar calcium for maximum activity, and has a restricted substrate specificity. It degraded native and denatured collagen more readily than casein, albumin, or fibronectin and denatured collagen (gelatin) was a better substrate than native collagen. The plasminogen activator released by neurons has a MW of 51 kDa and is converted to an active 32 kDa form. Its physiochemical properties are similar to urokinase and it was precipitated by a rabbit antiserum produced against human urokinase. A large fraction of both proteases was released by distal processes and/or growth cones suggesting that these proteases could be involved in growth cone functions. PMID- 2989046 TI - Plasmid vectors for the regulated, high level expression of eukaryotic genes in Escherichia coli. AB - A series of plasmid vectors has been constructed for the regulated, high level expression of foreign genes in E. coli. The vectors express cloned genes under the control of the tac promoter, which is a hybrid of trp and lac promoter sequences. Some of our expression vectors carry in addition to the tac promoter, the efficient lacZ ribosome binding site followed by unique cloning sites. These vectors can be used to express cloned genes directly, i.e. in an unfused, mature form. A second type of vector provides, in addition to the above regulatory elements, a translation initiation codon (ATG) for the expression of genes which have been isolated in an incomplete form (for example: cDNA). A third type of vector allow readily the construction of gene fusions to the E. coli beta galactosidase gene, which may stabilize otherwise unstable eukaryotic proteins, and thus allows the production of high amounts of specific antigens in E. coli. With the above vectors, several eukaryotic and viral proteins, including SV40 small tumor antigen, human fibroblast interferon and herpes simplex glycoproteins have been expressed. PMID- 2989048 TI - Towards the development of a new foot and mouth disease vaccine. AB - The causative agent of foot-and-mouth disease is a picornavirus with a single stranded positive sense RNA genome of about 8000 nucleotides. There are seven main serotypes of the virus and a large number of subtypes. Virus harvested from cultured cells, and chemically inactivated, forms the basis of modern vaccines. The present vaccines have successfully controlled virus outbreaks in endemically infected areas but are unstable and require refrigeration. Early studies have shown that the ability to stimulate neutralizing antibodies resides in only one of the four capsid proteins, VPl. Although a number of groups have attempted to produce subunit vaccines from VPl using genetic engineering techniques, VPl may not be sufficiently immunogenic for effective vaccination. More encouraging are the results that have been reported for synthetic peptides corresponding to defined regions of the VPl protein. Further development of these immunogens will be needed to produce a new vaccine that can compete in cost and effectiveness with the present FMD vaccines. PMID- 2989047 TI - Amplification and stability of recombinant plasmid molecules with a very large insert of foreign genetic material. AB - The capacity of bacterial plasmid vector pAT 153 for harbouring and for amplifying in a stable way a very large insert of foreign genetic material was screened. The DNA of a tree shrew herpesvirus (strain 2) with a molecular weight of about 130 megadalton (200 kilobasepairs) consisting of a long unique DNA sequence without direct and inverted repeats (greater than 40 basepairs), was used for molecular cloning. A successful insertion and stable amplification of a DNA fragment with 49 megadalton (74 kilobasepairs) cloned in Eco RI site of pAT 153 was performed, which resulted in the construction of recombinant DNA plasmid pTH2-E-C1. The stability of the recombinant plasmid pTH2-E-C1 with a super insert of 74 kilobasepairs (49 MD) indicates that the bacterial plasmid vector pAT 153 has really a high capacity for harbouring foreign genetic material. PMID- 2989049 TI - Development of molecular probes for simian herpesvirus detection. AB - A number of serologic procedures are available for specifically determining past infection due to H. hominis, or H. simiae (B virus). A herpesvirus isolated from primate tissues, however, does not lend itself to specific identification. Procedures currently in routine use simply do not provide an unequivocal identification of the isolate. We have previously demonstrated that H. simiae contain polypeptide and glycoprotein antigens distinct from those of other herpesviruses. At selected intervals, infected cells and controls were harvested after being both pulsed as well as continuously labelled from 4-24 hours post infection with 3H-glucosamine. 3H-mannose, and 3H-L-amino acid mixture. Between 40-45 infected cell polypeptides were observed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) with 7-11 glycoproteins noted. Analysis of the bands by electroblot utilizing anti-B virus sera showed the presence of eleven reactive bands within this molecular weight range from preparations in infected Vero and HEp-2 cell lines. Glucosamine labelled major glycoproteins migrated at 130K, 100K, 95K, 65K, 39K; mannose moieties migrated at 95K and 65K. SDS-PAGE profiles of B-virus when reacted with anti-gA/gB from HSV-1 and HSV-2 showed weak reactivities on the 125-130K complex. Anti-VP5 from HSV-1 failed to indicate any reactivity. Three polypeptides were revealed by SDS-hydroxylapatite analysis of the 130K complex with approximate molecular weights of 115K, 123K, and 130K; the 115K and 123K bands reacted with antisera prepared against gA/gB from HSV-1 and HSV-2, but not to anti-IgC or anti-2gC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989050 TI - "Cytoplasmic" islet cell antibodies. Evidence that the target antigen is a sialoglycoconjugate. AB - We have biochemically treated (periodate, borohydride, neuraminidase, organic solvents) frozen sections of human pancreas and studied the reactivity of islet cell-antibody-positive human sera and monoclonal antibodies. The autoantigen of pancreatic sections has the properties of sialic acid containing glycolipid. PMID- 2989051 TI - The effect of in vivo glucose administration on human erythrocyte Ca2+-ATPase activity and on enzyme responsiveness in vitro to thyroid hormone and calmodulin. AB - To characterize endogenous control mechanisms for human erythrocyte membrane Ca2+ ATPase ("calcium pump") activity, we studied the effect of changes in blood glucose concentration in vivo within the physiologic range on Ca2+-ATPase activity in red cells. Red cells obtained in the course of induced hyperglycemia were also studied to determine susceptibility of membrane Ca2+-ATPase to stimulation in vitro by thyroid hormone and calmodulin, both of which have been shown previously to enhance Ca2+-ATPase activity. Oral glucose administration (75 g) to eight healthy, adult subjects induced predictable increases in concentrations of blood glucose and immunoreactive insulin. Basal levels of activity of Ca2+-ATPase in red cells obtained after glucose ingestion fell 55% (P less than 0.025) by 30 min after glucose with recovery of enzyme activity to levels not significantly different from basal by 60 min. Activity of red cell Ca2+-ATPase at time zero was significantly stimulated in vitro by thyroxine (T4, 10(-10) M), triiodo-L-thyronine (T3, 10(-10) M), and calmodulin (100 ng/mg membrane protein). In vivo glucose administration led to depression of red cell enzyme responsiveness in vitro to T4 and T3; recovery from this effect did not occur by 120 min after oral administration of glucose. Calmodulin responsiveness of the enzyme in vitro was less significantly reduced in red cells obtained after glucose ingestion. Intravenous (i.v.) glucose administration (20 g) to five subjects also led to decreased basal enzyme activity (61% of fasting level at 20 min). A significant decrease in response of enzyme to T4 was achieved by 8 min after glucose administration (P less than 0.02), with recovery by 60 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989052 TI - Prophylaxis of constipation by wheat bran: a randomized study in hospitalized patients. AB - To evaluate the efficacy of wheat bran in preventing constipation, 200 hospitalized patients were randomly allocated to groups receiving either a dietary supplement of 40 g bran daily or no dietary supplement at all. A quarter of the bran group patients refused to take their bran from the very beginning (refusers), one third stopped bran consumption during the study (dropouts), and only 42% of the patients continued on bran until discharge or death (participants). Independent of a previous history of constipation, neither the hospital incidence of constipation nor the average percentage of days on laxatives was significantly different between the bran group and the control group. Only the dropouts were significantly more constipated than the control patients, whereas no such difference could be demonstrated in the refusers or participants. It is concluded that the administration of bran as a prophylactic laxative confers no benefit in patients hospitalized for a relatively short time. PMID- 2989054 TI - Characterization of the photolysis of 2,4,5,2',4',5'-hexabromobiphenyl. AB - 2,4,5,2',4',5'-Hexabromobiphenyl (2,4,5-HBB) was irradiated with ultraviolet light in hexane with stirring, and photolysis was monitored by gas chromatography (GC). 2,4,5-HBB decomposed at an average rate of 0.66 +/- 0.02 mumol/min and the reaction appeared zero order from 0.159 to 1.59 mM 2,4,5-HBB. Several polybrominated biphenyl (PBB) congeners were identified as photoproducts of 2,4,5 HBB. 2,4,5,2',5'-Pentabromobiphenyl (-PBB), formed by para debromination, accumulated at a higher rate than did 2,4,5,3',4'-PBB, formed by ortho debromination. 2,4,5,2',4'-PBB was formed by meta debromination. 3,4,3',4' Tetrabromobiphenyl (-TBB) was found as a secondary photoproduct, formed by ortho debromination of 2,4,5,3',4'-PBB. 2,5,2',5'-TBB and 2,4,2',5'-TBB were formed by debromination of 2,4,5,2',5'-PBB para and meta, respectively. 2,5,3',4'-TBB could be formed by either ortho debromination of 2,4,5,2',5'-PBB or para debromination of 2,4,5,3',4'-PBB. Rates of degradation and accumulation of the penta- and tetrabrominated biphenyls were also studied. The ultraviolet spectra of the 2,4,5 HBB photolysis mixture, as well as the purified components, were studied and are also reported. PMID- 2989053 TI - Immunotoxicologic evaluation of the respiratory system: animal models for immediate- and delayed-onset pulmonary hypersensitivity. AB - A wide variety of industrial materials is known to cause allergic pulmonary reactions. Respiratory symptoms may occur either immediately upon exposure to the agent (immediate-onset response), or several hours later (delayed-onset reaction). In order to determine both the mechanism of response, and a safe exposure level which would prevent development of pulmonary sensitization, animal models are currently being developed for both types of sensitivity responses. In the models, emphasis is placed on simulating conditions present in the industrial environment. For example, (1) exposure to agents is via the inhalation route, (2) animals are unrestrained and unanesthetized both during exposure and during elicitation of response, and (3) reactive chemicals, as opposed to hapten-protein conjugates, are used for exposure. In the model for immediate-onset sensitivity, concentration-response relationships have been observed between the concentration of agent inhaled and the percentage of animals becoming sensitized. Agents employed were bacterial subtilisin and toluene diisocyanate. Identification of "no observable effect" threshold concentrations implies that exposure levels can be proposed for industrial chemicals which will not result in sensitization of workers. Further development of the animal models, followed by calibration of the models to humans, should allow immunotoxicologic evaluation of chemicals for their respiratory sensitizing ability and recommendation of threshold-limit values (TLVs) which would prevent sensitization. PMID- 2989055 TI - Photolysis products of 2,4,5,2',4',5'-hexabromobiphenyl: hepatic microsomal enzyme induction and toxicity in Sprague-Dawley rats. AB - The irradiation of 2,4,5,2',4',5'-hexabromobiphenyl (2,4,5-HBB) by ultraviolet light created a mixture of lower brominated polybrominated biphenyl (PBB) congeners. Three photoproducts, 2,4,5,3',4'-pentabromobiphenyl (-PBB), 2,4,5,2',5'-PBB, and 3,4,3',4'-tetrabromobiphenyl (3,4-TBB), as well as 2,4,5-HBB and the photolyzed 2,4,5-HBB mixture, were administered to rats as a single ip injection (90 mg/kg, except 3,4-TBB, which was given at 2 mg/kg) 2 weeks before sacrifice. All treatments except 3,4-TBB induced NADPH-cytochrome P-450 reductase and aminopyrine-N-demethylase activities while all treatments except 2,4,5-HBB induced ethoxyresorufin-O-deethylase and UDP-glucuronosyltransferase activities. Thymus to body weight and spleen to body weight ratios were unchanged compared to controls for all treatments whereas an increase in the liver weights was observed for all treatment groups. Histologic examination revealed that the photolyzed 2,4,5-HBB mixture caused moderate to severe hepatocyte enlargement. Results of tissue analysis for the pure PBB congeners indicated that 2,4,5,2',5'-PBB and 3,4 TBB were metabolized in vivo and this was confirmed by in vitro metabolism studies. The results revealed that the photolyzed 2,4,5-HBB mixture caused a mixed-type induction of hepatic drug-metabolizing enzymes. This is most likely due to the effect of 2,4,5-HBB and toxic congeners formed during the irradiation of 2,4,5-HBB. 2,4,5,3',4'-PBB, which is toxic and apparently not metabolized, is believed to be the major congener contributing to the increased toxicity of the photolyzed 2,4,5-HBB mixture since 3,4-TBB was metabolized and appeared not to be as potent as inducer of aryl hydrocarbon hydroxylase activity. PMID- 2989056 TI - [Various norms of eubiotic feeding in healthy elderly persons]. PMID- 2989057 TI - Structure of the pX protein deduced from the nucleotide sequence of a cDNA clone of pX mRNA in cells infected with human T-cell leukemia virus type I. AB - A splice donor site of pX mRNA of human T-cell leukemia virus type I was elucidated by analyzing a cDNA clone of poly A+ RNA isolated from cat fibroblast cells infected with the virus. The donor site was located near the 5' end of the env gene. The putative N-terminal amino acid sequence of the pX protein was deduced to be Met-Ala-His---. PMID- 2989058 TI - Sera of adult T-cell leukemia patients and carriers of human T-cell leukemia virus type I in Japan do not cross-react with antigens of lymphadenopathy virus. AB - The immunological cross-reactivity of human T-cell leukemia virus type I (HTLV-I) related sera with lymphadenopathy virus (LAV) was investigated. The sera tested were obtained from 36 adult T-cell leukemia patients, 182 HTLV-I carriers, and 284 control individuals in Nagasaki. Two test methods were utilized: indirect immunofluorescence on LAV-producing cells and solid-phase radioimmunoassay with lysate of LAV. The results were exclusively negative for the presence of antibody activity against LAV, indicating the absence of cross-reaction and endemy in Nagasaki. PMID- 2989059 TI - Interferons inhibit syncytia-forming ability and in vitro transmission of human T cell leukemia virus. AB - Interferon(IFN)-alpha and -beta strongly inhibited syncytia formation of human T cell leukemia virus (HTLV). They also inhibited transmission of HTLV to normal human fibroblasts. These phenomena suggest a physiological role of IFNs in defense against HTLV infection. PMID- 2989060 TI - Hepatocellular carcinoma in the Philippines: a clinicopathological study of 150 cases. AB - Clinicopathological features were examined in 150 hepatocellular carcinoma cases autopsied from 1977 to 1983 in the Philippines and were compared to those of cases reported in Japan. The characteristics of hepatocellular carcinoma in the Philippines were as follows. 1. The age of autopsied patients with hepatocellular carcinoma in the Philippines was approximately 10 years younger than in Japan. The Philippines cases included even 10- to 11-year-old patients. 2. The macroscopic type of hepatocellular carcinoma in the Philippines consisted predominantly of the massive type of Eggel's classification. On the other hand, an encapsulated tumor was found in only one case. 3. The histological characteristics of hepatocellular carcinoma in the Philippines were not especially well defined. Hyaline globules and fatty metamorphosis were found in 1 and 12, respectively, of 90 Philippine hepatocellular carcinoma cases. 4. The frequency of HBs antigen in hepatocellular carcinoma in the Philippines was 37%. This is almost equal to that in Japan. PMID- 2989061 TI - Stimulation of calcium reabsorption observed in advanced breast cancer patients with hypercalcemia and multiple bone metastases. AB - In an attempt to analyze the pathological processes which lead to hypercalcemia in patients with multiple bone metastases, 23 advanced breast cancer patients with multiple bone metastases, three hypercalcemic patients with other malignancies and seven early breast cancer patients without any distant metastasis were studied. Of the 23 patients with advanced breast cancer, nine showed serum calcium levels higher than 10 mg/dl. In five of the nine hypercalcemic patients with advanced breast cancer, urinary cyclic AMP excretion was lower than 4 nmol/100 ml of glomerular filtrate (GF), indicating that the secretion of parathyroid hormone was suppressed. However, urinary cyclic AMP excretion was higher than 4 nmol/100 ml of GF in the other four hypercalcemic patients with advanced breast cancer and three hypercalcemic patients with other malignancies. In patients with higher urinary cyclic AMP excretion, fractional excretion of calcium (FECa) showed a negative correlation (r = 0.83, P less than 0.05) with urinary cyclic AMP. Parathyroid hormone immunoreactivity was not detected in any of six patients showing serum calcium levels higher than 11 mg/dl. These results suggest that in about a half of hypercalcemic patients with advanced breast cancer and multiple bone metastasis, there is a factor which increases urinary cyclic AMP and enhances calcium reabsorption in the kidney, but which is different from parathyroid hormone. This factor may facilitate retention of calcium mobilized into the circulation by bone metastases, and lead to hypercalcemia. PMID- 2989062 TI - Generation of lymphomagenic mouse type-C viruses from radiation- or chemically induced non-producer T-cell lymphoma cell lines infected with non-oncogenic ecotropic virus. AB - Highly lymphomagenic mouse type-C viruses were generated from radiation- or chemically-induced T-cell lymphoma cell lines of NFS/N mouse origin infected with a non-oncogenic ecotropic virus E4. By analysis of these progeny viruses, the following results were obtained. 1) The viruses were lymphomagenic in neonatally inoculated NFS/N and C3H/He mice and W/Fu rats but not in Balb/c and C57BL/6N mice, indicating that they possess the Fv-1n tropism of exogenously infected parent virus. 2) Lymphomagenic viruses consisted of plural viral subpopulations. Recombinant mink cell focus-inducing (MCF) and ecotropic viruses were cloned from them. Inoculation of either MCF or ecotropic virus alone or both viruses together did not cause lymphoma in NFS/N mice and there was no evidence of viral replication in the recipients. 3) Inoculation of either MCF- or ecotropic virus infected NFS-ME cells alone did not cause lymphoma development in pre-irradiated NFS/N mice, while transplantation of both MCF- and ecotropic virus-infected NFS ME cells resulted in the development of lymphomas of host origin. These results show that lymphomagenic MCF virus was generated through the recombination of E4 viral genome and a modified proviral DNA of endogenous viruses present in radiation- or chemically-induced lymphomas, and that an interaction or synergism of MCF and ecotropic viruses is required for MCF virus to exert lymphomagenic activity. PMID- 2989063 TI - Induction of functional differentiation of a human monocytic leukemia cell line (THP-1) by retinoic acid and cholera toxin. AB - The human monocytic leukemia cell line, THP-1, is induced to differentiate into more functionally mature monocyte (macrophage)-like cells by incubation with retinoic acid at concentrations of 10nM or higher. There is no apparent morphological change accompanying this functional maturation. These induced cells show increases in nitroblue tetrazolium reduction, immunoerythrophagocytosis, hexose monophosphate shunt activity, and 5'-nucleotidase and NAD+-glycohydrolase activities. Prostaglandin E2, dibutyryl cyclic adenosine 3':5'-monophosphate, or T-lymphocyte-derived differentiation-inducing activity, all inactive or less active alone, increase the extent of differentiation of THP-1 in combination with 10nM retinoic acid. THP-1 is also induced to differentiate by 0.1nM or higher concentrations of cholera toxin. Furthermore, 24,24-difluoro-1 alpha,25 dihydroxyvitamin D3 induces less differentiation of THP-1 compared to retinoic acid. Dimethyl sulfoxide and 12-O-tetradecanoylphorbol-13-acetate show no induction of functional differentiation. THP-1 thus joins the list of leukemic myelomonocytic cell lines (e.g., the promyelocytic HL-60 and the monoblast-like U 937) that are blocked at a relatively late stage of maturation and which differentiate in response to retinoic acid. PMID- 2989064 TI - Establishment and characterization of Ph1-positive and Ph1-negative lymphoblastoid cell lines from a patient with chronic myelogenous leukemia. AB - Two continuously growing in vito cell lines, PB-1049 and LN-1049, were established from a patient with Ph1-positive chronic myelogenous leukemia in extramedullary blastic crisis. PB-1049 was established from Epstein-Barr virus (EBV)-infected peripheral lymphocytes of the patient and had a 46,XY,t(9;22) karyotype. This cell line was shown to be tumorigenic in nude mice, and cultured cells recovered from the tumor nodule showed a 46,XY,6p+,t(9;22) stem line karyotype. LN-1049 was derived from a lymph node culture of the same patient, and was found to be non-tumorigenic in nude mice and karyotypically normal. Immunological examinations on EBV-induced antigens, surface and cytoplasmic immunoglobulin, and monoclonal antibodies, and some enzymatic and electron microscopic studies revealed that both cell lines had attained differentiation into late-B cell stage. PMID- 2989066 TI - [Surgical resection of hepatocellular carcinoma. Retrospective study of 42 cases]. AB - We studied the results 42 liver resections performed for hepatocellular carcinoma. Thirty-three patients underwent major liver resections. Hepatocellular carcinomas were associated with cirrhosis in 30.9 p. 100 of cases. The overall operative mortality was 24.3 p. 100: 60 p. 100 in patients with cirrhosis who underwent major liver resections and 12.5 p. 100 in patients with cirrhosis undergoing limited resections. The overall 5-year survival rate was 14.8 p. 100; the 5-year survival rate in patients with hepatocellular carcinoma without cirrhosis was 20 p. 100 whereas no patient with associated cirrhosis of the liver survived at 3 years. We conclude that surgical resection of hepatocellular carcinoma must be performed whenever possible: a limited resection should be done if the tumor is small, especially in patients with cirrhosis; a major liver resection must be proposed when the tumor is large and if the remaining liver parenchyma is normal. PMID- 2989065 TI - Strain differences in the antitumor activity of an immunopotentiator, Nocardia rubra cell-wall skeleton, in B10 congenic and recombinant mice. AB - An immunogenetic evaluation of the antitumor activity of the immunopotentiator Nocardia rubra cell-wall skeleton (N. rubra-CWS) has been performed using non virus-producing tumors induced by the Schmidt-Ruppin strain of Rous sarcoma virus (RSV) in B10 congenic and recombinant mice. Live tumor cells mixed with either N. rubra-CWS or a placebo control were inoculated intradermally into the right flank of syngeneic mice. With N. rubra-CWS, the development and growth of B10 mouse tumors, S1018(B10) and B10SA2F, in B10(H-2b) mice were completely inhibited in all test mice. B10.A(5R) mouse tumor, S322(5R), was completely suppressed in 23 of 25 B10.A(5R)(H-2i5) mice, and B10.BR mouse tumor, S623(BR), was completely suppressed in 7 of 14 B10.BR(H-2k) mice. However, B10.D2 mouse tumor, S908(D2), in B10.D2(H-2d) mice and B10.A mouse tumors, S826(BA) and B10ABr1F, in B10.A(H 2a) mice were not suppressed at all by N. rubra-CWS. N. rubra-CWS showed a remarkable antitumor effect in B10 mice, not only in mixed inoculation with the tumor cells but also with intratumoral administration. These effects were not seen in B10.A mice. Peritoneal exudate cells (PEC) from B10 and B10.A strains previously treated intraperitoneally with N. rubra-CWS and/or MMC-treated tumor cells showed cytolytic and cytostatic activities on tumor cells derived from both strains. These results did not reflect the strain difference seen in the in vivo studies. Mitogenic activity of N. rubra-CWS on spleen cells, however, was significantly different among B10 congenic mice; N. rubra-CWS induced greater blastogenesis of spleen cells from B10 mice than of those from B10.A mice, corresponding to the in vivo results showing strain difference. These results suggest that the host's immunogenetic background may play an important role in cancer therapy with immunopotentiators. PMID- 2989067 TI - [Fulminating hepatitis due to suloctidil]. PMID- 2989068 TI - [Hepatitis B virus, serological markers of viral infections and humoral immunity in alcoholic cirrhosis]. AB - In order to define the role of hepatitis B virus (HBV) in alcoholic liver disease and to study the relationship between HBV and other common viruses, the serological markers of viral disease (HBV, Rubella, Polio, Herpes, and Cytomegalovirus-CMV) were compared in 163 patients with alcoholic cirrhosis (group C), 100 patients with alcoholic steatosis (group S) and in 168 non alcoholic control subjects (group NA). A significantly increased prevalence of HBV markers in group C was related to the presence of anti-HBc antibodies, in 10.5 p. 100 of cirrhotic patients, vs. 1.2 p. 100 in group S and 1 p. 100 in group NA (p less than 0.01). In cirrhotic patients with HBV markers (HBV +) incidence of alcoholic hepatitis was 4 times lower and the total duration of alcohol overconsumption was significantly lower than in cirrhotic patients without these markers (HBV-). Hepatic function tests were not different in HBV + and HBV- cirrhotic patients, excepted for the ASAT/ALAT ratio (1.55 +/- 0.10 vs. 1.92 +/- 0.12; p less than 0.05). Prevalence of anti-CMV antibodies, and anti herpes greater than 1/100 antibodies, was significantly increased in S and C groups (p less than 0.01). Anti-Rubella, Polio, and CMV antibody titers were higher (p less than 0.05) in HBV + than in HBV- cirrhotic patients. In cirrhotic subjects, titers of these 3 anti-virus antibodies were not related to alcoholic hepatitis or to IgG and IgM concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989069 TI - [Hepatocellular carcinoma in France. Clinical, biological and virological aspects in 197 patients]. AB - The authors report the clinical and biological features in 197 patients with hepatocellular carcinoma seen in two French hospitals. Mean age was 63 +/- 12 years. Eighty-nine per cent were men. Cirrhosis was present in 88 p. 100. Alcoholic liver disease was associated with hepatocellular carcinoma in 71.5 p. 100. At the time of diagnosis, ascites was present in 62 p. 100 of the patients, jaundice in 49 p. 100, encephalopathy in 20 p. 100 and gastrointestinal bleeding in 12.5 p. 100. Twenty patients (10 p. 100) did not have any of these complications. An increase in serum gammaglutamyl transpeptidase and ASAT was the most frequent biological abnormality observed in 96 and 94 p. 100 of patients respectively. Hypercalcemia and a high hematocrit were present in 5 and 6 p. 100 of patients respectively. Serum HBs Ag (RIA) was present in 17.5 p. 100 of patients, anti-HBc in 50 p. 100 and anti-HBs in 33.5 p. 100; 38.5 p. 100 of patients had no serum HBV marker. Serum alphafetoprotein levels were higher than 20 ng/ml, 250 ng/ml and 1,000 ng/ml in 76.5 p. 100, 43.5 p. 100 and 33 p. 100 of patients respectively. There were no relationships between the presence of serum markers of HBV or high alphafetoprotein levels and clinical and biological data. These results confirm that the clinical, biological and virological aspects of hepatocellular carcinoma in France are similar to those reported in other western countries. PMID- 2989070 TI - [Effects of a tamoxifen-norethisterone combination in 16 patients with hepatocellular carcinoma]. PMID- 2989071 TI - Opisthorchis viverrini infection and cholangiocarcinoma. A prospective, case controlled study. AB - During a prospective, 24-mo case-controlled study, 551 patients from northeastern Thailand were independently evaluated for Opisthorchis viverrini infection, hepatobiliary tract disease, and hepatic carcinoma to determine whether there was any association between hepatic carcinoma and O. viverrini infection. Stool examination by the formalin-ether concentration method revealed O. viverrini ova in 389 (70.6%) patients. Of the 551 patients, 72 (13.1%) had both clinical and laboratory evidence of hepatobiliary tract disease, chronic liver disease, or hepatic carcinoma, alone or in combination. Of these 72 patients, 28 (38.9%) had a liver biopsy that revealed cholangiocarcinoma in 7 patients with O. viverrini ova in their stools, and in 4 patients without. In another patient with ova in the stool combined hepatocellular carcinoma and cholangiocarcinoma was found. In the 4 patients with cholangiocarcinoma who had no O. viverrini ova in their stools, ova were detected in the bile fluid aspirated from the intrahepatic biliary tree during exploratory laparotomy. An additional patient with clinically suspected cholangiocarcinoma and O. viverrini ova in stool had a left supraclavicular lymph node biopsy specimen taken that revealed metastatic adenocarcinoma; this adenocarcinoma was interpreted as compatible with cholangiocarcinoma. Cholangiocarcinoma, therefore, was found only in patients with O. viverrini ova in stool or in the intrahepatic biliary tree. Statistical analysis revealed that patients with known O. viverrini infection had a higher incidence of cholangiocarcinoma than did patients without such infection (X2 test, p less than 0.05). PMID- 2989072 TI - Benefit of transcatheter arterial embolization for ruptured hepatocellular carcinoma complicating liver cirrhosis. AB - In 6 patients with spontaneous rupture of hepatocellular carcinoma complicating liver cirrhosis, but with no occlusion of the main portal trunk, transcatheter arterial embolization was performed within 7 days of the rupture. All 6 patients were thought to be inoperable because of shock state or severe hepatic dysfunction. In all 6 patients, the progressive decrease in the hematocrit ceased soon after the embolization. Five patients survived for 31-168 days after the embolization; 1 patient who developed septicemia died 10 days later. We conclude that transcatheter arterial embolization is beneficial as a procedure of first choice for ruptured hepatocellular carcinoma when the portal blood flow is maintained. PMID- 2989073 TI - Naloxone is protective against indomethacin-induced intestinal ulceration in the rat. AB - Naloxone, an opiate antagonist, was reported to protect against stress ulcers in dogs and rats. We studied its possible protective effect against indomethacin induced intestinal ulceration in the rat. Naloxone was indeed found to possess a marked protective effect on the intestinal mucosa (ulcer index 73.3 +/- 13.6 vs. 273.8 +/- 21.8, p less than 0.001). Naloxone was found to elevate basal intestinal mucosal prostaglandin E2 (p less than 0.001) and cyclic adenosine monophosphate levels (p less than 0.005) but was unable to overcome the inhibition of prostaglandin E2 caused by indomethacin. An increase of cyclic adenosine monophosphate levels was seen, however, even in the presence of indomethacin, suggesting that cyclic adenosine monophosphate, but not prostaglandins, may play a role in the protective effect of naloxone. PMID- 2989074 TI - Gender differences in antacid-induced phosphate deprivation in rats. AB - To determine whether an animal model could be used to study the susceptibility of women to antacid-induced phosphate deficiency, 6-wk-old male and female rats were given basic aluminum carbonate gel (Basaljel) (1 ml/100 g body wt) or distilled water by gastric intubation daily for 3 wk. Rats were fed either ad libitum (group 1) or by pair-feeding (group 2) with pelleted rat food containing 0.74% phosphorus. In group 1, baseline, 1-wk, and 3-wk values for serum phosphorus in Basaljel-treated females were 7.7 +/- 0.2, 6.3 +/- 0.2, and 6.2 +/- 0.2 mg/dl, respectively. Corresponding values for control females were 7.8 +/- 0.3, 7.0 +/- 0.2, and 7.3 +/- 0.2 mg/dl. Values for treated females were significantly lower (p less than 0.02) than values for control females by 1 wk of treatment. Basaljel treated males did not differ from controls. The pattern for group 2 was similar. Intestinal absorption and intramuscular stores of phosphate were assessed in group 1. After 3 wk of treatment, [32P]phosphate assimilation from the duodenum into the body was lower in Basaljel-treated females than in controls (33.8% +/- 1.9% vs. 49.8% +/- 6.2% of the luminally administered dose, p less than 0.05). This was due to increased retention of [32P]phosphate in the intestine of treated females (19.9% +/- 2.0% vs. 11.9% +/- 2.4% for control females, p less than 0.02). Results in jejunum were similar. Total intramuscular phosphate in females was significantly lower (p less than 0.005) than in males both before and after antacid treatment. Thus hypophosphatemia in the female rat during antacid administration is probably secondary to the additive effects of decreased assimilation and decreased soft tissue stores of phosphate. PMID- 2989075 TI - Colorectal carcinoma in patients with Crohn's disease. AB - Epidemiologic studies have shown a fourfold to 20-fold increased risk of colorectal carcinoma in patients with Crohn's disease as compared with the general population, but management for this risk is controversial. This paper presents a series of 10 patients with Crohn's disease and colorectal carcinoma from one institution and a review of the literature. Compared with 118 consecutive patients having colorectal carcinoma of the usual type at the same institution, the 10 patients were younger (mean age 55 vs. 65 yr, p less than 0.05) and had a higher prevalence of mucinous carcinoma (50% vs. 9%, p less than 0.01), providing evidence in addition to the previous epidemiologic results that Crohn's disease is a predisposing (premalignant) condition for colorectal carcinoma. Eight of the 10 patients had an antecedent history of Crohn's disease, ranging from 4 to 51 yr (mean 24 yr); 2 patients presented with colorectal carcinoma and were found to have Crohn's disease. Of particular note, 9 of the 10 patients had colitis or ileocolitis, and carcinoma occurred in bypassed rectum in 2 patients. One patient had two carcinomas. Three of the 11 carcinomas were not recognized preoperatively. The anatomic sites of the cancers were not significantly different from usual colorectal carcinoma, with 7 of the 11 tumors (64%) in the sigmoid colon and rectum. Dysplasia was identified in all 10 patients, and all of the 10 resected carcinomas were found to be contiguous with high-grade dysplasia. The findings in this study support the proposals based upon epidemiologic data that surveillance should be considered for patients with clinically evident colorectal involvement by Crohn's disease or a bypassed segment of colorectum. PMID- 2989076 TI - [Detection of various papilloma virus types in cytologic smears of precancerous conditions and cancers of the uterine cervix]. AB - DNA-in situ hybridisation on epithelial cells taken from cervical swabs of 101 different patients cytologically diagnosed as III D to V (Munich classification 1975) were performed using 32P-labelled DNA of the human papilloma virus (HPV) types 6, 11, 16 and 18 as smears. The following correlations between the cytological classification and DNA hybridisation were obtained: 55 out of 58 women (94.8%) having a normal Pap smear (Pap II) were negative by hybridisation as well, three cases contained HPV 6 or 11.59.3% of patients revealing abnormalities characteristic of a papillomavirus infection reacted with the 32P labelled DNA, while HPV 6/11 or 16/18, respectively, occurred at approximately the same rate of frequency. In cases with severe dysplasia, carcinoma in situ or invasive cervical carcinoma (IVa, IVb, V) HPV 16 or 18 were more frequently found (67%) than in materials with mild dysplasia (IIID; 32% positive). HPV 6 or 11, on the other hand, was present only once in nine IVb/V-cases (11%), in 17% of IVa cases, but in 32% of patients with mild dysplasia. Infections with all different virus types were found in 9% of the IVa-und 16% of the IIID-cases. The high prevalence of HPV 16 or 18 in CIS as well as in invasive carcinomas is in line with the biological correlation between both types of lesions. HPV 6 or 11 is more frequently associated with the usually reversible mild dysplastic epithelial alterations. The DNA-in situ swab hybridisation should, therefore, be of diagnostic value and might help to optimise the therapy for the individual patient. PMID- 2989077 TI - [Diagnostic problems of virus-induced atypical squamous cell proliferations in the area of the anus and genitalia. A case report on Bowenoid papulosis]. AB - Papillomatous proliferations of squamous epithelium of the genital and anal region termed Bowenoid Papulosis (BP) macroscopically resemble condylomata acuminata, whereas histologically they show in part severe dysplasias. Despite the similarity of these lesions to Bowen's disease they are - like condylomas - induced by papilloma viruses and may regress spontaneously. Condyloma-like but predominantly flat proliferations of squamous epithelium of the transitional zone between exocervix and endocervix, which are likewise caused by viruses and have a prognosis similar to that of BP, are even more problematic in differential diagnostic differentiation from cervical carcinoma in situ. Both these atypical proliferations of squamous epithelium of the exocervix referred to as "flat condylomas" and BP contain koilocytes as a sign of cell regression relatively typical of virus-induced cell change. In a reported case of a 38-year old woman with widespread BP of the ano-genital region intranuclear papilloma virus antigen could be demonstrated immunohistochemically in the papillomatous proliferations of squamous epithelium and furthermore in sections made from the filed paraffin blocks of a cone biopsy taken five years ago. In two of altogether seven cases of BP presented in tables papilloma virus antigen could be shown as well. In one woman of already 54 years of age widespread BP is described more comprehensively, because due to the feature of inverted papillomatosis carcinoma was suspected clinically. BP and flat condylomas have been defined as terms only in recent years and - as virus-induced lesions - been differentiated from carcinoma in situ on account of a more favourable prognosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989078 TI - [Dystrophy and precancerous changes in the vulva: symptoms, histology and therapy]. AB - The article reports about the relatively rare diseases kraurosis vulvae (lichen sclerosus et atrophicus) and leucoplakia vulvae and the symptoms, aetiology, histology and therapy of these diseases. Both diseases are considered to be a "dystrophy" since 1976, by international convention. This publication is a report on 168 patients treated during the last 20 years (1964-1983). 34 patients suffered from diabetes mellitus (21%); 73.2% of the women were aged between the 61st and 90th year of life. 15 women already had an incipient vulvar carcinoma besides kraurosis (8.9%). Precancerous alterations, such as morbus Bowen (29 cases), morbus Paget of the vulva (6 cases), and erythroplasia of Queyrat are subsequently discussed. PMID- 2989079 TI - [Bafverstedt lymphadenosis benigna cutis. Differential diagnostic delineation from Paget disease]. AB - A case is reported showing differential diagnostic delineation between Paget's disease and Bafverstedt's disease. Both show single localisation and an eczematoid appearance of the nipple or mamillary area. PMID- 2989080 TI - [Chemiluminescence of blood and bone marrow cells in children with leukemia]. PMID- 2989081 TI - Effects of L-thyroxine and ovine growth hormone on smoltification of amago salmon (Oncorhynchus rhodurus). AB - In a study of the hormonal control of salmon smoltification, L-thyroxine (T4; 200 ppm in diet), ovine growth hormone (oGH; 2 micrograms/g intramuscular injection weekly), or a combination of these hormones was administered to underyearling amago salmon parr for 72 days. Administration of both T4 + oGH remarkably stimulated growth of the fish. All hormone treatments caused body silvering of the fish. The body silvering was the greatest in the group treated with T4 + oGH. Both T4 + oGH and oGH increased survival rate of the fish in 32% seawater, and kept plasma osmolarity and plasma sodium concentration lower than those of T4 treated and control groups in 27% seawater. Significant elevation of gill Na+, K+ ATPase activity was found in T4 + oGH-treated fish. On the other hand, T4 by itself did not affect the seawater tolerance of the fish. These findings suggest that GH or synergistic action of GH and T4 plays an important role in seawater adaptation during smoltification of amago salmon. PMID- 2989082 TI - Reevaluation of the relative activities of the pituitary glycoprotein hormones (follicle-stimulating hormone, luteinizing hormone, and thyrotrophin) from the green sea turtle, Chelonia mydas. AB - The discovery that the follicle-stimulating hormone (FSH) previously prepared from the green sea turtle, Chelonia mydas, contained a major neurohypophysial contaminant prompted a repurification and characterization of the glycoprotein hormones in this turtle. Results reaffirmed the physicochemical distinctiveness of the three hormones. Minimal cross-contamination between hormones (less than 2%) was achieved by ion-exchange chromatography, subunit dissociation (of contaminating luteinizing hormone (LH], gel filtration, and immuno-affinity chromatography. New preparations of FSH and thyrotrophin (TSH) derived from adult pituitaries proved to be more potent than those described previously (the degree depending on the nature of the assay); FSH showed the expected increase in activity based on estimated contamination of previous preparations. LH was similar to original preparations except for enhanced activity in FSH radioreceptor assays. Binding assays (in heterologous and homologous systems) again demonstrated the general absence of an FSH-specific receptor in the reptilian (chelonian and squamate) testes. In an in vivo bioassay in the lizard Anolis, the turtle FSH was orders of magnitude more potent than LH in stimulating both testis growth and androgen secretion, but in vitro LH was considerably more potent than FSH in stimulating androgen secretion in squamate and chelonian testes. Thus, the possibility exists that androgen secretion in some chelonian systems may exhibit a high degree of LH specificity like that of mammals and birds. PMID- 2989083 TI - Kinetic studies on the enzymes involved in estrogen biosynthesis and evidence for existence of a single 5-Ene-3beta-hydroxysteroid dehydrogenase complex in the ovary of the freshwater catfish, Clarias batrachus. AB - Substrate velocity kinetics and other characteristics of two key enzymes involved in estrogen biosynthesis in the ovary of the catfish, Clarias batrachus, were studied. Enzyme reactions were measured spectrophotometrically during the initial phase of vitellogenesis. 5-Ene-3beta-hydroxysteroid dehydrogenase (5-ene-3beta HSD) was found to have a different Michaelis-Menten constant (Km) for each of three 5-ene-3beta-hydroxysteroids, (pregnenolone (P5), 17alpha hydroxypregnenolone (17alpha-P5), and dehydroepiandrosterone (DHA) but the constant for 17beta-hydroxysteroid dehydrogenase (17beta-HSD) was similar for both the substrates androstendione (A-dione) and estradiol-17beta (E2). 5-Ene 3beta-HSD required exclusively NAD as cofactor for the conversion of P5, 17alpha P5, and DHA. A-dione was converted into testosterone (T) by a NADP-dependent 17beta-HSD, whereas there was an obligatory requirement of NAD for the conversion of E2 to estrone (E1). P5 was utilized more efficiently as substrate by 5-ene 3beta-HSD than DHA, as indicated by its lower Km/Vmax ratio. The rate of the combined substrate reaction was less than the sum of the rate of reactions measured separately for each of the three sets of substrate pairs. These results indicate that a single enzyme is responsible for the oxidation of three types of 5-ene-3beta-hydroxysteroids studied. PMID- 2989084 TI - Molecular cloning of alpha-amylase genes from Drosophila melanogaster. II. Clone organization and verification. AB - Restriction maps of an alpha-amylase structural gene clone, lambda Dm65, and of four putative alpha-amylase pseudogene clones are presented. Two alpha-amylase structural genes, inverted with respect to each other, are contained in lambda Dm65. Subregions of internal DNA sequence homology within lambda Dm65 and of cross-homology between the presumptive pseudogene clones and lambda Dm65 were determined. Subregions of cross-homology between the Drosophila clones and the mouse alpha-amylase cDNA clone, pMSa104, were also determined. The presence of functional alpha-amylase structural genes in lambda Dm65 was verified by injection of appropriate subclones into the germinal vesicle of Xenopus oocytes, followed by incubation of the oocytes under conditions that allowed coupled transcription and translation of injected genes to occur. Subclones of the 3.8- and 5.6-kb EcoRI fragments of lambda Dm65 were shown to code for alpha-amylase isozymes 1 and 3, respectively, of Drosophila melanogaster Canton-S. Both subclones are homologous to RNA of a size sufficient to accommodate the alpha amylase-coding information. No RNA species homologous to other subcloned EcoRI fragments of lambda Dm65 was detected. PMID- 2989085 TI - [Organization of the genes responsible for colicin Ib synthesis and immunity to it in plasmid ColIb-P9]. AB - To study the localization and expression of the ColIb-P9 plasmid genes responsible for colicin Ib synthesis and immunity to it, we isolated a series of Tn5 insertion mutants of recombinant plasmid pIV41 containing the colicin Ib gene in EcoRI fragment of ColIb-P9 (2.7 kb) and the deletion plasmid carrying only a part of the colicin gene. The direction of colicin Ib gene transcription was determined by the analysis of the polypeptides synthesized in minicells carrying the mutant plasmids. The pIV41 plasmid containing cells have been shown to be resistant to both colicin Ib and Ia activities. This type of resistance is usually associated with chromosomal mutations resulting in loss of cell receptors for colicin Ib adsorption. Apparently, the EcoRI fragment of ColIb-P9 studied contains no gene responsible for immunity to colicin. It has been shown that this gene is a portion of SalI fragment (22 kb) of the ColIb-P9, the fragment also carrying the gene which mediates synthesis of colicin Ib. PMID- 2989086 TI - The genetics of hormone-induced cyclic AMP production and phospholipid N methylation in inbred strains of mice. PMID- 2989087 TI - Genome organization of the Pseudomonas aeruginosa narrow host range plasmid R91-5 determined by deletion and cloning analysis. PMID- 2989088 TI - Cloning of the human myoglobin gene. AB - Genomic clones that contain the human myoglobin gene were isolated by cross hybridization to the porcine myoglobin cDNA from human genomic libraries. The myoglobin gene is about 10.5 kb long and contains two introns as in the case with hemoglobin genes. Diverse tandem repetitive sequences with 45 FokI sites are located 1100-1750 bp upstream from the putative cap site of myoglobin mRNA. Two kinds of direct repeats (116 and 58 bp) are contained in this repetitive region. In addition, a purine-rich sequence starting from the FokI site is found around 68-114 bp upstream of a putative initiation site of transcription. Another 33-bp sequence is repeated four times in the first intron and bordered by a 9-bp direct repeat, a structure that is common with the eukaryotic transposable elements. PMID- 2989089 TI - Construction of a human X-chromosome-enriched phage library which facilitates analysis of specific loci. AB - A human X-chromosome-enriched MboI-partial-digest recombinant library in phage lambda Charon30 has been constructed. Twelve out of the thirteen X-chromosome DNA sequences that were tested were present in the library. Most regions were covered in overlapping phage inserts; mean insert size was 13.7 kb. One phage from the library allowed detection of a 225-bp insertion of DNA into a region near the Duchenne muscular dystrophy (DMD) locus. Another recombinant phage represents an expansion of a region which exhibits extensive and varying homology with other human chromosomes, including the Y, as well as with rodent DNA. The present library should have widespread use for examining DNA sequences on the human X chromosome. PMID- 2989090 TI - Nucleotide sequence and transcription of a gene encoding human tRNAGlyCCC. AB - A phage lambda clone containing a 13.1-kb human DNA fragment was isolated and found to contain a tRNA gene encoding a glycine tRNA. The nucleotide sequence of the gene and its flanking regions has been determined. The gene does not have an intervening sequence nor does it encode the 3'-terminal CCA sequence found in mature tRNAs. Although this tRNA gene has an anticodon sequence of CCC, it has a striking homology (96%) with a human glycine tRNA which has an anticodon of GCC. As in other eukaryotic tRNA genes, the coding region contains a characteristic internal split promoter sequence, and the 3'-flanking region has a typical RNA polymerase III termination site of five consecutive T residues. There is no apparent sequence in the 5'-flanking region which could serve as a regulatory element. This gene is accurately transcribed in vitro by RNA polymerase III using a HeLa cell-free system. During the course of in vitro transcription, larger precursor tRNAGlyCCC transcripts are processed to yield a mature-sized tRNA product. A precursor-product relationship was established by comparing the ribonuclease A fingerprints of the precursor and product tRNA transcripts. PMID- 2989091 TI - Isolation and characterization of Chinese hamster ribosomal DNA clones. AB - We have examined the ribosomal RNA (rRNA) genes of the Chinese hamster ovary (CHO) cell line. A partial EcoRI library of genomic CHO DNA was prepared using lambda Charon-4A. We isolated two recombinants containing the region transcribed as 45S pre-rRNA and 13 kb of external spacer flanking 5' and 3' to the transcribed region. These sequences show restriction site homology with the vast majority of the genomic sequences complementary to rRNA. In addition to this form of rDNA, Southern blot analysis of EcoRI-cut CHO genomic DNA reveals numerous minor fragments ranging from 2 to 19 kb which are complementary to 18S rRNA. We isolated one clone which contains the 18S rRNA gene and sequences 5' which appear to contain length heterogeneity within the non-transcribed spacer region. We have nine additional cloned EcoRI fragments in which the homology with 18S rRNA is limited to a 0.9-kb EcoRI-HindIII fragment. This EcoRI-HindIII fragment is present in each of the cloned EcoRI fragments, and is flanked on both sides by apparently nonribosomal sequences which bear little restriction site homology with each other or the major cloned rDNA repeat. PMID- 2989092 TI - Transposon insertion mutagenesis of Pseudomonas aeruginosa with a Tn5 derivative: application to physical mapping of the arc gene cluster. AB - For insertional mutagenesis of Pseudomonas aeruginosa, a derivative of the kanamycin-resistance (KmR) transposon Tn5 was constructed (Tn5-751) that carried the trimethoprim-resistance (TpR) determinant from plasmid R751 as an additional marker. Double selection for KmR and TpR avoided the isolation of spontaneous aminoglycoside-resistant mutants which occur at high frequencies in P. aeruginosa. As a delivery system for the recombinant transposon, plasmid pME305, a derivative of the broad-host-range plasma RP1, proved effective; pME305 is temperature-sensitive at 43 degrees C for maintenance in Escherichia coli and P. aeruginosa and deleted for IS21 and the KmR and primase genes. In matings with an E. coli donor carrying pME9(= pME305::Tn5-751), transposon insertion mutants of P. aeruginosa PAO were recovered at approx. 5 X 10(-7)/donor at 43 degrees C. Among Tn5-751 insertional mutants 0.9% were auxotrophs. A thr::Tn5-751 mutation near the recA-like locus rec-102 is useful for the construction of recombination deficient strains. Several arc::Tn5-751 mutants could be isolated that were defective in anaerobic utilization of arginine as an energy source. From three of these mutants the arc gene region was cloned into an E. coli vector plasmid. Since Tn5-751 has a single EcoRI site between the TpR and KmR genes, EcoRI generated fragments carrying either resistance determinant plus adjacent chromosomal DNA could be selected separately in E. coli. Thus, a restriction map of the arc region was constructed and verified by hybridization experiments. The arc genes were tightly clustered, confirming earlier genetic evidence. PMID- 2989093 TI - Cloning of the DNA repair genes mtcA, mtcB, uvsC, uvsD, uvsE and the leuB gene from Deinococcus radiodurans. AB - A gene library from Deinococcus radiodurans has been constructed in the cosmid pJBFH. A 51.5-kb hybrid cosmid, pUE40, that transduced Escherichia coli HB101 from leucine dependence to independence was selected, and a 6.9-kb fragment which carried the leuB gene from D. radiodurans was subcloned into the EcoRI site of pAT153. The DNA repair genes mtcA, mtcB, uvsC, uvsD and uvsE, which code for two D. radiodurans UV endonucleases were identified by transforming appropriate repair-deficient mutants of D. radiodurans to repair proficiency with DNA derived from the gene library. Hybrid cosmid pUE50 (37.9 kb) containing an insert carrying both the mtcA and mtcB genes was selected and 5.6- and 2.7-kb DNA fragments carrying mtcA and mtcB, respectively, i.e., the genes that code for UV endonuclease alpha, were subcloned into the EcoRI site of pAT153. The three genes uvsC, uvsD and uvsE, that code for UV endonuclease beta, were all present in the 46.0-kb hybrid cosmid pUE60. The uvsE gene in a 12.2-kb fragment was subcloned into the HindIII site of pAT153 and the size of the insert reduced to 6.1 kb by deletion of a 6.7-kb fragment from the hybrid plasmid pUE62. None of the uvs genes introduced into the UV-sensitive E. coli CSR603 (uvrA-) was able to complement its repair defect. The mtcA, uvsC, uvsD and uvsE genes were found in the 52.5-kb hybrid cosmid pUE70. It is concluded that the DNA repair genes mtcA, mtcB, uvsC, uvsD and uvsE are located within an 83.0-kb fragment of the D. radiodurans genome. PMID- 2989094 TI - Orientation and expression of the cloned hemolysin gene of Pseudomonas aeruginosa. AB - The structural gene for Pseudomonas aeruginosa hemolysin, carried on recombinant plasmid pSL2 and cloned in Escherichia coli, was analyzed by insertional and deletional mutagenesis. Expression of the hemolysin was blocked by insertion of transposon Tn5 into different locations. Two of the mutants allowed detectable synthesis of truncated hemolysin polypeptides of two different sizes and thus defined the structural gene. The location of the hemolysin gene in the recombinant plasmid, and the direction of transcription, were further established by nuclease BAL 31 digestion, and by construction of gene fusions between hemolysin and beta-galactosidase. Evidently, the tet promoter contributed to the majority of the expression of cloned hemolysin gene, but the Pseudomonas promoter was present in the cloned DNA and was functional in E. coli since inactivation of the tet promoter either by Tn5 insertion or by deletion decreased synthesis of the 80-kDal hemolysin but did not fully abolish it. PMID- 2989095 TI - A plasmid cloning system utilizing replication and packaging functions of the filamentous bacteriophage fd. AB - DNA cloning vectors were developed which utilize the replication origin (ori) of bacteriophage fd for their propagation. These vectors depend on the expression of viral gene 2 that was inserted into phage lambda, which in turn was integrated into the host genome. The constitutive expression of gene 2 in the host cells is sufficient for the propagation of at least 100 pfd plasmids per cell. In addition to the fd ori, the pfd vectors carry various antibiotic-resistance genes and unique restriction sites. Some of these vectors have no homologies to commonly used pBR plasmids or to lambda DNA. The nucleotide sequence of the vectors can be deduced from published sequences. Large DNA inserts can be stably propagated in pfd vectors; these are more stable than similar DNA fragments cloned in intact genomes of filamentous bacteriophage. Inclusion of phage sequences required for efficient phage packaging and infection with a helper phage resulted in formation of phage particles containing single-stranded plasmid genomes. Growth at 42 degrees C without selective pressure results in loss of pfd plasmids. PMID- 2989096 TI - Nucleotide sequence of a major class-III phage-T3 RNA-polymerase promoter located at 98.0% of phage-T3 genetic map. AB - The entire nucleotide sequence of a 409-bp HincII fragment, located within the MboI-E fragment on bacteriophage T3 DNA and containing a major class-III T3 RNA polymerase promoter positioned at 98% on the standard T3 genetic map, has been determined. Alignment of this class-III promoter with previously determined T3 RNA polymerase promoters, with start points of transcription (+1) in register, indicates high degree of sequence conservation between position -16 to +6 among all T3 RNA polymerase promoters. The conserved portion of the (-) strand sequence is 5'-A-TA-T-AT-A-C-C-C-T-C-A-C-T-A-A-A-G-G-G-A---3'. This fragment also contains an open reading frame (ORF) with a translational start codon located at position +146 which is preceded by a potential ribosome binding site (RBS). There is more than 70% amino acid-sequence homology between the deduced sequences of the -NH2 terminal region of this putative T3 phage protein and the corresponding protein coded by bacteriophage T7 (protein of T7 gene 19.5). PMID- 2989097 TI - Purification of a new restriction endonuclease, StyI, from Escherichia coli carrying the hsd+ miniplasmid. AB - A new restriction endonuclease, StyI, free of contaminating nuclease activities, has been isolated from Escherichia coli carrying the hsd+ miniplasmid of Salmonella typhi origin. In the presence of 10 mM Mg2+, it recognizes and cleaves a hexanucleotide sequence of 5'-C decreases C(AT)(AT)GG. The advantages of the StyI endonuclease include its stability, high yield (more than 2 X 10(3) units/g of wet cells), easy handling of producer cells, and the ability to recognize new sequences, CCAAGG and CCTTGG. PMID- 2989098 TI - The DNA between Rz and cosR in bacteriophage lambda is nonessential. AB - Near the right end of phage lambda DNA, between gene Rz and the cos site, are 2050 bp of apparently non-coding DNA. We have cloned a lambda DNA fragment containing this DNA into a plasmid and constructed a deletion, omega l, extending from a site within the Rz gene to a site about 560 bp from cos. This deletion could be recombined into viable lambda phage at a frequency equal to that observed for the undeleted sequence. Recombinant phage lambda carrying the omega l deletion were demonstrated to have the same burst size and kinetics of phage production as undeleted lambda. The omega l deletion can be used to extend the capacity of lambda cloning vectors and to provide a region for the insertion of heterologous DNA which should exhibit controllable high level expression from the lambda late promoter, p'R. PMID- 2989099 TI - A novel deletion found during cloning of a synthetic palindromic DNA. AB - A 212-bp palindromic DNA comprising two copies of the left end of bacteriophage Mu was assembled from chemically synthesized oligonucleotides and inserted into plasmid pUC9. When cloned and propagated in Escherichia coli, the palindrome was found to be unstable and was generally lost. However, in a few cases, a precise, asymmetric deletion of one half of the insert was observed. This pattern of deletion suggests that the symmetry axis region of the palindrome was involved as recognition site in the deletion process. PMID- 2989100 TI - The araBAD operon of Salmonella typhimurium LT2. I. Nucleotide sequence of araB and primary structure of its product, ribulokinase. AB - Hybrid plasmids containing the araBAD operon of Salmonella typhimurium LT2 were characterized by Southern blot and genetic analyses. The nucleotide sequence of araB was determined. The araB gene product, ribulokinase (EC 2.7.1.16), was purified and the results of amino acid composition analysis and partial amino acid sequence are in agreement with predictions from the DNA sequence. Ribulokinase is 569 amino acid residues long and has a calculated Mr of 61 793. Ribulokinase shares significant homology with xylulose kinase from Escherichia coli. Codon usage in the araB gene does not favor those codons which have intermediate codon-anticodon binding energy. PMID- 2989101 TI - IS50-mediated inverse transposition: specificity and precision. AB - The IS50 elements, which are present as inverted repeats in the kanamycin resistance transposon, Tn5, can move in unison carrying with them any interstitial DNA segment. In consequence, DNA molecules such as a lambda::Tn5 phage genome are composed of two overlapping transposons - the kan segment bracketed by IS50 elements (Tn5), and lambda bracketed by IS50 elements. During direct transposition, mediated by IS50 "O" (outside) ends, the kan gene is moved and the lambda vector is left behind. During inverse transposition, mediated by the "I" (inside) ends of the IS50 elements, the lambda vector segment is moved and the kan gene is left behind. Direct transposition is several orders of magnitude more frequent than inverse transposition (Isberg and Syvanen, 1981; Sasakawa and Berg, 1982). We assessed the specificity and precision of the rare events mediated by pairs of I ends by mapping and sequencing independent inverse transpositions from a lambda::Tn5 phage into the amp and tet genes of plasmid pBR322. Using restriction analyses, 32 and 40 distinct sites of insertion were found among 46 and 72 independent inverse transpositions into the amp and tet genes, respectively. Eleven sites were used in two or more insertion events, and the two sites in tet used most frequently corresponded to major hotspots for the insertion of the Tn5 (by direct transposition). The sequences of 22 sites of inverse transposition (including each of the sites used more than once) were determined, in eleven cases by analyzing both pBR322-IS50 junctions, and in eleven others by sequencing one junction. The sequence of the "I" end of IS50 was preserved and 9-bp target sequence duplications were present in every case analyzed. GC pairs were found at each end of the target sequence duplication in ten of the eleven sites used more than once, and also in seven of the other eleven sites. Our data indicate that transposition mediated by pairs of "I" ends is similar in its specificity and precision to the more frequent transposition mediated by IS50 "O" ends. PMID- 2989102 TI - The promoter of the long terminal repeat of feline leukemia virus is effective for expression of a mouse H-2 histocompatibility gene in mouse and human cells. AB - DNA-mediated gene transfer techniques have been used to study the effectiveness of a novel construction involving the feline leukemia virus long terminal repeat (FeLV LTR) for expressing the mouse H-2 Ld gene in mouse and human cells. In this construction, the transcription initiation (promoter) and termination (polyadenylation) functions of the FeLV LTR have been split by insertion of a promoterless H-2 gene between them. An S1 nuclease assay has been developed that makes it possible to measure accumulated LdRNA against a background of endogenous major histocompatibility antigen RNAs in mouse and human cells. In mouse cells, the H-2 Ld gene was expressed at approximately equal levels (measured as accumulated RNA) when driven either by its own promoter or by the FeLV LTR construction. In human cells, expression at the RNA level was highest when driven by the FeLV LTR. We conclude that the FeLV LTR construction is useful for expressing foreign genes in human cells. PMID- 2989103 TI - A direct-selection vector derived from pColE3-CA38 and adapted for foreign gene expression. AB - The construction of a plasmid vector, pVT25, which allows an efficient and direct selection for transformed cells carrying recombinant plasmids is described. In this vector, the replicon and ApR gene from plasmid pBR327 are fused to the colE3 gene of pColE3-CA38, whereby positive selection is based on the inactivation of the lethal colicin E3 by the insertion of a foreign DNA fragment. However, pVT25 can be maintained within the Escherichia coli cells when complemented with another plasmid, pVT26, which expresses the colicin E3 immunity (imm) and the TcR phenotypes. Furthermore, pVT25 was used to regulate the expression of the synthetic human proinsulin gene fused to the colE3 gene at the single ClaI site. The production of the characteristic C-peptide of proinsulin, monitored by radioimmunoassay, was shown to be under the control of the inducible promoter of the colE3 gene. PMID- 2989105 TI - Cloning of cDNAs coding for rat hepatic microsomal UDP-glucuronyltransferases. AB - Radioiodinated, affinity-purified, anti-UDP-glucuronyltransferase (UDPGT) antibodies have been used to isolate cDNAs coding for UDPGT(s) from a rat liver cDNA library cloned in the expression vector bacteriophage lambda gt11. The sizes of ten cloned cDNAs range from 0.3-2.1 kb. The identity of the cDNAs was confirmed by the hybrid-select translation and immunochemical analyses. Restriction mapping indicates that two classes of cDNA coding for different UDPGT mRNAs have been cloned. PMID- 2989104 TI - Cloning and expression of genes for lysozyme and a 20-kDal protein of colitis bacteriophage. AB - BamHI fragments of colitis phage DNA were cloned in pBR322 DNA, and the recombinant clones carrying the lysozyme gene were identified by lysozyme activity. The inserted DNA was 1.2 kb long and when expressed in minicells it produced lysozyme and a 20-kDal protein. Colitis-phage-specific mRNAs which hybridized to the insert were 0.5 kb and 0.7 kb long and were translated into lysozyme and a 20-kDal protein, respectively, in a cell-free system derived from rice embryos. They were transcribed as monocistronic mRNAs using the internal promoters present on the inserted DNA. PMID- 2989106 TI - Isolation of a mouse DNA fragment with homology to a Drosophila ribosomal protein gene. AB - A mouse genomic library in lambda Charon 4A was screened for putative ribosomal protein genes using a fragment of the gene encoding Drosophila ribosomal protein 49 as a hybridization probe under nonstringent hybridization conditions. A recombinant phage was selected and its restriction enzyme map determined. The major species of mouse poly(A)+ mRNA homologous to the putative gene is about 740 nucleotides long. PMID- 2989107 TI - Cointegrates carrying two copies of a Tn3 derivative in an inverted orientation. AB - We constructed a mutant of Tn3, Tn3 #2, that contains a 55-bp direct repeat of sequences near the amino-terminal coding region of the transposase, and an 8-bp EcoRI linker. This mutant transposase is functional. The plasmid carrying Tn3 #2, pMB8::Tn3 #2, recombines with the plasmid pHS1 at a frequency of 2.8 X 10(-7) recombinants per division cycle. This is similar to the recombination frequency of pHS1 and pMB8::Tn3+ (wild-type) which is 4.5 X 10(-6) recombinants per division cycle. One-third of the recombinants between pMB8::Tn3 #2 and pHS1 were approx. 22 kb in length. Restriction analysis and nucleotide sequencing showed that these large plasmids were Tn3 #2-mediated cointegrates formed by integration of pMB8::Tn3 #2 into pHS1. However, unlike Tn3 tnpR- -mediated cointegrates that contain direct repeats of the incoming element, Tn3 #2-mediated cointegrates carry two copies of Tn3 #2 in the form of inverted repeats. Like the tnpR- repeats, the Tn3 #2 repeats occur at both junctions between the parental plasmids, and are associated with a 5-bp direct duplication of the pHS1 target site. Furthermore, these recombinants contain a small deletion starting precisely at the end of Tn3 #2 and extending into pMB8 sequences. We propose a model for the generation of Tn3 #2-mediated cointegrates. PMID- 2989108 TI - Nucleotide sequence coding for the insecticidal fragment of the Bacillus thuringiensis crystal protein. AB - The insecticidal crystal protein (ICP) gene, icp, from a 68-kb plasmid derived from Bacillus thuringiensis subsp. sotto was cloned in Escherichia coli. The icp expression in E. coli cells was confirmed by both immunological and insect toxicity assays of the cell extract. The entire icp gene resides in the 6.6-kb PstI fragment, which codes for a 144-kDal peptide identical to the intact ICP, as determined by its size and reaction with anti-ICP antibody. Deletion analysis further revealed that the 2.8-kb region within the 6.6-kb PstI fragment codes for ICP. Analysis of the nucleotide sequence indicated that a peptide of 934 amino acid residues truncated at the C-terminal end is encoded by this 2.8-kb fragment. A unique feature of this truncated ICP is the abundance of cysteine and lysine residues within its C-terminal region. PMID- 2989109 TI - Mercuric reductase structural genes from plasmid R100 and transposon Tn501: functional domains of the enzyme. AB - The nucleotide sequence for the 2240 bp of plasmid R100 following the merC gene of the mercuric resistance operon has been determined and compared with the homologous sequence of transposon Tn501. The sequences following merC and preceding the next structural gene merA are unrelated between R100 and Tn501 and differ in length, with 72 bp in Tn501 and 509 bp in R100. The R100 sequence has a potential open reading frame (ORF) for a 140 amino acid polypeptide with a reasonable translational start signal preceding it. The merA genes contain 1686 (Tn501) and 1695 (R100) bp respectively. When optimally aligned, the merA sequences differ in 18% of their positions. These differences were clustered in specific regions. In addition, there was one nucleotide triplet in the Tn501 sequence which has no counterpart in the R100 sequence and one dodecyl-nucleotide sequence in the R100 sequence without counterpart in Tn501. Thus the predicted merA polypeptide of Tn501 contains 561 amino acids and the R100 counterpart contains 564 amino acids. Comparison of the R100 mercuric reductase sequences with that for human glutathione reductase [Krauth-Siegel et al.: Eur. J. Biochem. 121 (1982) 259-267], for which there is a 2 A resolution electron density map [Thieme et al.: J. Mol. Biol. 152 (1981) 763-782] shows a strong homology, with 26% identical amino acids and many conservative substitutions. This homology allows the conclusion that the active site of these enzymes and the contact positions for flavin adenine dinucleotide (FAD) and NADPH are highly conserved, while the amino- and carboxyl-terminal sequences differ. PMID- 2989110 TI - Transcriptional regulation of the MET3 gene of Saccharomyces cerevisiae. AB - The MET3 gene, coding for ATP sulfurylase (ATPS), an enzyme implicated in methionine biosynthesis in Saccharomyces cerevisiae, was cloned by functional complementation, after transformation, of a yeast met3 mutant strain. The cloned MET3 gene was used as a probe to measure the specific MET3 messenger RNA in a wild-type strain grown under conditions which promote or fail to promote repression of ATPS synthesis. It was found that the level of MET3 messenger RNA is reduced ten-fold when the strain is grown under conditions where ATPS synthesis is repressed, suggesting that the MET3 expression is regulated transcriptionally. The direction of transcription and the size of the transcript have been determined. PMID- 2989111 TI - New derivatives of the Streptomyces temperate phage phi C31 useful for the cloning and functional analysis of Streptomyces DNA. AB - The thiostrepton resistance gene (tsr) of Streptomyces azureus, and a synthetic oligonucleotide adapter sequence, were introduced into the DNA of attP-site deleted phage phi C31-based cloning vectors. The DNA of two of the new derivatives, KC515 and KC516, contains single sites for the enzymes BamHI, BglII, PstI, PvuII, SstI (two sites close together) and XhoI, available for the insertion of DNA of up to 4 kb. The two vectors also contain a cloned, promoterless viomycin phosphotransferase gene (vph) from Streptomyces vinaceus. When an internal segment of the Streptomyces coelicolor glycerol (gyl) operon was inserted at the appropriate position and in the correct orientation next to vph, it could bring about in vivo recombination leading to fusion of vph of the chromosomally located gyl operon, resulting in glycerol-regulated expression of viomycin resistance. Two other new phi C31 derivatives, KC505 and KC518, are PstI and BamHI replacement vectors, respectively, for 2-8-kb DNA fragments, and allow simple screening for the presence of inserted DNA. PMID- 2989113 TI - Nucleotide sequence of the streptokinase gene from Streptococcus equisimilis H46A. AB - The entire nucleotide sequence of a cloned 2568-bp PstI fragment from the genome of Streptococcus equisimilis H46A encoding the streptokinase gene (skc) has been determined. The longest open reading frame comprises 1320 bp which code for streptokinase. The protein is synthesized with a 26-amino acid residue N-terminal extension having properties characteristic of a signal peptide. Comparison of the deduced amino acid sequence with the available amino acid sequence of a commercial streptokinase reveals minor primary structure differences. The nucleotide sequencing of skc does not support the hypothesis that the gene has evolved by duplication and fusion, as suggested by internal twofold amino acid homologies of its product. Furthermore, the skc gene sequence shows no extended regions homologous to the staphylokinase gene. Upstream from the skc gene, the putative skc promoter and the ribosome-binding site sequence have been identified; downstream from the coding region, inverted repeat sequences thought to function as transcription terminators have been detected. PMID- 2989112 TI - Cloning of the hexA mismatch-repair gene of Streptococcus pneumoniae and identification of the product. AB - The hexA mismatch repair gene of Streptococcus pneumoniae has been cloned into multicopy plasmid vectors. The cloned hexA gene is expressed as judged from its ability to complement various chromosomal hexA- alleles. Its direction of transcription was defined and the functional limits were localized by original methods relying on homology-dependent integration of nonautonomous chimeric plasmids carrying chromosomal inserts into the chromosome. Comparison of the proteins encoded by recombinant plasmids and by restriction fragments allowed us to identify an Mr 94 000 protein as the probable product of the hexA gene. PMID- 2989115 TI - Cloning of a multicopy plasmid from the actinorhizad nitrogen-fixing bacterium Frankia sp. and determination of its restriction map. AB - An 8.3-kb multicopy plasmid, pFQ31, from the nitrogen-fixing Frankia sp. strain ArI3, was cloned into Escherichia coli plasmid vectors and analysed physically. pFQ31 has no detectable sequence homology with another Frankia plasmid, pFQ32, which is present in the same host. Derivatives of pFQ31 with an antibotic resistance marker were introduced into Streptomyces lividans, which is taxonomically related to Frankia, but no stable replication could be achieved. PMID- 2989114 TI - Vectors with restriction-site banks. III. Escherichia coli-Saccharomyces cerevisiae shuttle vectors. AB - The bank of unique restriction sites present in plasmid pJRD158 has been incorporated into new vectors carrying selective markers and replicons derived from commonly used Escherichia coli-Saccharomyces cerevisiae shuttle vectors pJDB207 and YRp7. The new vectors pMH158 and pJO158 have 21 and 23 unique restriction sites, respectively, and their complete DNA sequences are known. PMID- 2989116 TI - Nutritional needs of the elderly: debate and recommendations. AB - Despite all that has been written, little evidence supports the notion that the American diet for the elderly needs major modifications (table 3). Particularly in counseling and assessing the elderly, physicians must keep in mind that whatever technique older patients used to reach their present age is probably better than what we can recommend. In the last analysis, the most sensible nutritional recommendations we can make are to maintain ideal body weight, reduce animal protein, increase complex carbohydrates and fiber, reduce saturated fats and cholesterol, increase calcium intake, and avoid high doses of supplementary nutrients. If available data are any indication, many community-dwelling elderly persons are now making most of these modifications. PMID- 2989117 TI - [Hygienic basis for redistribution of chemical and biological pollutants in water]. PMID- 2989118 TI - [Affinity of enteroviruses and bacteriophages to natural aluminum silicate sorbents]. PMID- 2989119 TI - [Bioluminescent determination of ATP in peritoneal macrophages phagocytosing dust particles]. PMID- 2989120 TI - [Morphological assessment of pleural tumors induced in rats by Dzhetygara chrysotile asbestos and lizardite]. PMID- 2989121 TI - Constipation; common but preventable. PMID- 2989122 TI - A special recipe to banish constipation. PMID- 2989123 TI - Ovarian metastases from cancer of the lung: problems in interpretation--a report of seven cases. AB - Seven women, 26 to 66 (average 42) years of age, from whom ovarian tumors were removed before (three cases), synchronously with (three cases), or less than 1 year after (one case) the discovery of a pulmonary neoplasm are the subjects of this report. The ovarian tumor was unilateral in six cases and bilateral in one case and was unassociated with intraabdominal spread. The lung tumors, which were similar to the ovarian tumors on microscopic examination, were central in six cases and peripheral in one; three of them were small cell undifferentiated carcinomas, two large cell undifferentiated carcinomas, one a poorly differentiated adenocarcinoma, and one an atypical spindle cell carcinoid tumor. In each case the question of the primary site of the tumor had been raised clinically. The histological similarity of the ovarian and pulmonary tumors in all the cases to well-recognized forms of pulmonary neoplasia as well as additional clinical and pathological features of the cases led to an interpretation of a pulmonary origin of the tumors in all the cases. PMID- 2989124 TI - [Interdisciplinary cooperation with tumor after-care clinics]. PMID- 2989125 TI - [Myeloperoxidase deficiency: prevalence in Brescia Province and a study of microbicidal activity in granulocytes]. PMID- 2989126 TI - Automated differential white cell count (flow cytometry) in a case of eosinophilic total myeloperoxidase (MPO) deficiency. PMID- 2989127 TI - [Differential diagnosis, operative procedure and prognosis in complicated syndactylies]. AB - Two cases of congenital syndactyly of all fingers are presented. Special anatomical abnormalities found during the operations are discussed. The correlation with an anatomically and ontogenetically well-founded classification provides knowledge for use in the individual case which may be helpful in selecting the proper surgical procedure, in establishing a schedule, and in providing the prognosis. PMID- 2989128 TI - [Ingestion of button-batteries by children: the conservative approach]. PMID- 2989129 TI - [The role of hepatitis B virus infection in the development of primary hepatocellular carcinoma]. PMID- 2989130 TI - [Dopamine-induced up-regulation of alpha 2-adrenoceptors in crude synaptic membranes of rat brain]. AB - After cerebral cortical membranes were incubated with 0.1-100 microM of dopamine (DA) in 50 mM Tris-HCl buffer (pH 7.7) at 37 degrees C for 30 min, [3H] clonidine binding to alpha 2-receptors was increased in a concentration-dependent manner without changing [3H] WB4101 and [3H] DHA binding to alpha 1- and beta-receptors, respectively. Scatchard analysis of [3H] clonidine binding to cortical membranes showed that DA increased the Bmax in both high- and low-affinity components. The increasing effect of DA on [3H] clonidine was dependent on incubation time and temperature, and it was antagonized by pimozide and cis-flupenthixol. The addition of GTP produced a reduction in DA-induced elevation in [3H] clonidine binding, while that of cyclic AMP did not affect the effect of DA. DA and Mn2+, though both of them increased [3H] clonidine binding, appeared to act at a different site in the membrane. Furthermore, the DA-induced increase in [3H] clonidine was found uniformly in membranes prepared from 7 other regions of the rat brain. These results suggested that DA regulates specifically alpha 2 receptor density by stimulating D1-receptors and/or via other mechanism(s). PMID- 2989131 TI - [Pharmacological studies on schizandra fruits. III. Effects of wuweizisu C, a lignan component of schizandra fruits, on experimental liver injuries in rats]. AB - The effects of wuweizisu C, a lignan component of schizandra fruits, on liver injuries induced by carbon tetrachloride (CCl4), d-galactosamine and dl-ethionine were investigated by means of serum-biochemical and histopathological examinations in rats. Pretreatment or combined administration of wuweizisu C dose dependently reduced the elevation of serum transaminase activity and histological changes such as fatty degeneration, cell necrosis, inflammatory cell infiltration, etc., which were caused by the single administration of 1 ml/kg, p.o., or the repeated administration of 0.2 ml/kg, s.c., daily for 4 days of CCl4, respectively. The effects of wuweizisu C on the liver injuries induced by a low dose (200 mg/kg, i.p.) and a high dose (400 mg/kg, i.p.) of d-galactosamine were compared with those of uridine. Wuweizisu C significantly lowered the rise of serum transaminase activity after the administration of a low dose of d galactosamine in the serum-biochemical analysis. A tendency was also shown to inhibit cell necrosis and inflammatory cell infiltration caused by both doses of d-galactosamine in the histopathological examination. On the other hand, uridine markedly repaired the serum-biochemical and histopathological changes after the administrations of both doses of d-galactosamine. Also wuweizisu C cured the liver injury by the repeated administration of 150 mg/kg, i.p., daily for 4 days of d-galactosamine. After the repeated administration of 250 mg/kg, s.c., daily for 4 days of ethionine, liver cell atrophy, diffuse fatty degeneration and decrease of serum triglyceride were observed, but not cell necrosis. Wuweizisu C dose-dependently inhibited fatty degeneration and decrease of serum triglyceride. These findings suggest that wuweizisu C can be protective and/or therapeutic on hepatocellular phenomena such as cell necrosis, fatty degeneration, inflammatory cell infiltration, etc., in human hepatitis. PMID- 2989132 TI - [Behavioral pharmacology of amantadine with special references to the effect on abnormal behavior in mice and rats]. AB - Behavioral effects of amantadine, especially on abnormal behavior in mice and rats, were reevaluated, in comparison with those of tricyclic antidepressants, methamphetamine and L-DOPA. 1) Amantadine at 10 approximately 50 mg/kg, i.p., tended to decrease ambulation and rearing in rats and mice. The drug at 50 mg/kg, i.p., caused piloerection and hyperirritability and at doses over 80 mg/kg, i.p., it impaired coordinated motor activity in rats. 2) Amantadine inhibited methamphetamine-induced hyperactivity, but apomorphine-induced stereotyped behavior was unaffected in rats. 3) Amantadine was equipotent to imipramine in suppressing haloperidol-induced catalepsy in rats, but L-DOPA was without effect. On the other hand, amantadine was 40 and 400 times as potent as imipramine and L DOPA, respectively, in suppressing delta 9-tetrahydrocannabinol (THC)-induced catalepsy in rats. 4) Amantadine was as potent as imipramine in suppressing the muricide of olfactory bulbectomized rats, but was 3.5, 8.8 and 225.5 times as potent as methamphetamine, imipramine and L-DOPA, respectively, in inhibiting THC induced reversible muricide in rats. 5) Amantadine at 50 mg/kg did not elicit circling behavior in the rat with unilateral nigral lesion induced by 6 hydroxydopamine. 6) Amantadine at high doses caused irritable aggression characterized by squealing in rats pretreated with intraventricular 6 hydroxydopamine. The most important characteristic of amantadine is its prominent effect suppressing the THC-induced catalepsy and muricide. This may be a reflection of the feature of amantadine activating the dopaminergic as well as the serotonergic systems. PMID- 2989134 TI - Toluene diisocyanate and lung function. PMID- 2989133 TI - Relationship of body-weight gain to longevity and to risk of development of nephropathy and neoplasia in Sprague-Dawley rats. AB - The relationship of weight gain to survival, risk of development of chronic progressive nephropathy and risk of development of various neoplasms has been studied in the control groups from two routine chronic toxicity studies in Sprague-Dawley rats. The groups comprised 100 CFY strain rats of each sex observed up to the age of 109 wk and 120 CD strain rats of each sex observed up to 111 wk of age (females) or 121 wk (males). The eventual incidence of tumours was found to be related to body weight at several ages. There was also a statistically significant association between high body weight at various ages and increased mortality, particularly in the CD strain and particularly in females. The 'heavy' rats proved to have an increased risk of developing both progressive nephropathy and certain tumours. This relationship was particularly marked for pituitary tumours in both sexes and for benign and malignant mammary tumours in females, and was significant irrespective of whether tumours coexisting with marked or severe progressive nephropathy were classified as fatal or incidental. There was also some evidence that increased body weight was positively associated with risk of islet-cell tumours and lipomatous tumours in males and fibromatous tumours in females. The observations illustrate how non specific factors, such as those that affect body weight, may profoundly influence mortality and tumour incidence in chronic toxicity studies. The findings also highlight the difficulty of classifying particular neoplasms as incidental or fatal where other potentially life-threatening pathology (e.g. progressive nephropathy) is present. PMID- 2989135 TI - Angiotoxicity in swine of a moderate excess of dietary vitamin D3. AB - The effects of five different dietary levels of vitamin D3 on the coronary arteries of groups of 17-60 2-month old weanling Yorkshire swine were studied. Four groups were fed the following levels of vitamin D3 per ton of ration continuously for 4 months: group I--100,000 IU, group II--300,000 IU, group IV- 2,000,000, and group V--4,000,000 IU. Swine in group III were fed the diet containing 100,000 IU of vitamin D3 per ton of ration for the first 2 months of the study after which they were fed a diet supplemented with 4,000,000 IU of vitamin D3 per ton of ration for the remaining 2 months of the study. The highest degree of intimal thickening of the coronary arteries was observed among group V. The thickened areas contained numerous lipid-containing cells and degenerate cells without stainable lipid. Electron microscopic examination revealed a greater frequency of degenerate cells without stainable lipid in the coronary arteries of groups III and IV than in groups I and II. These results suggest a possible link between excessive daily intake of vitamin D3 and the development of human coronary atherosclerosis. PMID- 2989136 TI - [The effect of phosphodiesterase inhibitors on retinal function of the eye undergoing arterial perfusion]. PMID- 2989137 TI - [Medial tarsal tunnel syndrome. A not infrequent, but often mistaken, nerve compression syndrome]. PMID- 2989138 TI - The influence of dietary fiber on gastric transit time. AB - Little is known about the effect of dietary fiber on gastric emptying. At present, we do not know whether any therapeutic effects result from delayed gastric transit time. Functional scintigraphy was used to determine the effect of wheat bran, guar and pectin on the transit time of 200 ml fruit juice and 200 or 400 ml of a liquid test meal in normal healthy subjects. The transit time of fruit juice was significantly delayed by the addition of 4 g guar (viscosity 7079 mPA X s). Neither wheat bran, pectin nor guar resulted in a delay of the test meal transit time despite maximal increases in viscosity of up to 21,500 mPa X s. We therefore conclude that dietary fiber does not significantly delay the gastric transit time of test meals regardless of the increase in viscosity. The reduction in postprandial glucose levels after ingestion of guar or pectin is, therefore, probably not due to delayed gastric emptying. PMID- 2989139 TI - Effects of short-term exposure to catecholestrogens on serum concentrations of gonadotrophins and metabolism of catecholestrogens in ovariectomized rats. AB - Equimolar amounts (36.8 nmol) of estradiol and the two catecholestrogens (CE's) 4 hydroxyestradiol (4-OHE2) and 2-hydroxyestradiol (2-OHE2) were injected subcutaneously to ovariectomized adult rats. Serum levels of both LH and FSH were determined at short-term intervals. Moreover, serum levels of the administered steroids and their main free and conjugated metabolites were monitored. Serum levels of the injected steroids reached peak values at different time points: estradiol between 60 and 240 min, CE's between 30 and 60 min. Peak height also differed significantly: with estradiol highest (1500 pg/ml), followed by 4-OHE2 (540 pg/ml) and then 2-OHE2 (135 pg/ml) (ratio 11:4:1). This mirrored the different MCR's of the steroids: CE's, especially 2-OHE2, were rapidly and extensively methylated and/or - to a lesser degree - conjugated. Estradiol remained mainly unchanged. LH-serum levels in steroid treated animals showed - irrespective of the steroid used - a uniform reaction pattern: they were significantly depressed 60 to 240 min after injection and - with the exception of estradiol treated rats - reached pretreatment levels again 480 min after injection. Ultra-short (15 min) effects were not observed. FSH serum levels in CE treated animals were not significantly altered, only E2 application led to a significant but small decrease in FSH levels 240 and 480 min after its injection. In conclusion, neither the effect of 4-OHE2 nor that of 2-OHE2 corresponded to the different MCR's or the MCR-corrected affinities for the classical estrogen receptor. A non-genomic mechanism may be responsible for this impaired effect of CE's. PMID- 2989140 TI - Dissociation of adrenal androgen and cortisol levels in acute stress. AB - Patients recovering from acute surgical stress often excrete increased 17-OH corticosteroids with no change in 17-ketosteroids. The explanation for these findings is unclear. In order to investigate possible divergence between cortisol and adrenal androgen metabolism in acute stress, repeated morning cortisol and dehydroepiandrosterone (DHA) measurements were made in patients undergoing ACTH stimulation 48 to 96 hours preoperatively, followed by determinations before and during major surgery, also performed in the morning. Cortisol and DHA are largely metabolized by the liver, so liver blood flow under a constant general anesthetic regimen known not to affect cortisol metabolism was monitored by pre- and intraoperative indocyanine green dye clearance. Results indicated no difference between the cortisol and DHA stimulation resulting from two hours of ACTH stimulation or major surgery, and a small (14.4%) decline in hepatic blood flow during general anesthesia. However, while DHA concentrations remained constant immediately preceding surgery, cortisol concentrations increased by 61% (P less than 0.05). Previous studies have also demonstrated increased concentrations of cortisol before surgical procedures, presumably due to psychological stress. However, this is the first demonstration of a dissociation between concentrations of cortisol and an adrenal androgen due to psychological stress. PMID- 2989141 TI - Amino acid uptake and protein synthesis in rat testes: stimulation by dissociable factors. AB - Low concentration (25 mM) of sodium in the incubation medium produced a decrease in the amino acid uptake by the testis tissue as well as a reduction in the response to FSH. In this experimental condition, the basal protein synthesis and the stimulatory effect of FSH was not modified. The subcutaneous administration of testosterone to 15 day old rats increased the protein synthesis in the testis without any modification in the amino acid uptake. The addition of DBcAMP (1 mM) or glucose (14 mM) to the incubation medium increased the protein synthesis in the testes of immature (12 day-old) or prepubertal (32 day-old) rats respectively. The amino acid uptake was not modified. In immature rat testes, with protein synthesis completely inhibited by cycloheximide, the restoration of the sodium concentration in the incubation medium to normal levels produced an increase in amino acid uptake. The results above seem to indicate that protein synthesis and amino acid uptake in rat testes tissue can be regulated, at least partially, by different factors. PMID- 2989142 TI - The mechanism(s) of the enhancement of renal compensatory hypertrophy by ACTH in the rat. PMID- 2989143 TI - Immunohistochemical examination of pituitary adenomas. Comparison to clinical and endocrinological findings. AB - A comparison was made with the data of 62 cases of pituitary adenoma, evaluated pre- and postoperatively, including as well the results of immunohistochemical hormone examination (also for calcitonin). Prolactin was found in 18 of the 21 adenomas carrying the preoperative diagnosis of prolactinoma, whereas cells containing other hormones (growth hormone, LH, FSH, TSH, ACTH, beta-endorphin), were only occasionally present. The growth hormone was strongly positive in the adenoma tissue in 16 of the 17 cases of acromegaly. 5 of these adenomas were accompanied by a marked hyperprolactinemia and also contained many prolactin cells. 6 of the 19 adenomas diagnosed as being 'inactive' contained hormone positive cells, but only a very small number of cells. ACTH was found in 3 of the 4 pituitary adenomas of patients with Cushing's disease. 2 of these were also positive for beta-endorphin. The tissue of 1 gonadotrophic adenoma (with elevated FSH in serum) gave positive results with an anti-LH antiserum. Calcitonin was not found in any adenoma. The preoperative serum prolactin levels did not quantitatively correlate with the percentage of prolactin-positive cells. PMID- 2989144 TI - Inhibition of the ACTH adrenal response to stress by treatment with hydrocortisone, prednisolone and dexamethasone in the rat. AB - 16- and 4-week-old intact and adrenalectomized rats have been treated with different doses of the three glucocorticoids hydrocortisone, prednisolone and dexamethasone by gavage. The delayed feedback effect on plasma ACTH and corticosterone response to an ether stress have been assessed. Almost complete suppression of corticosterone response 20 min after an ether stress and an ACTH suppression to 20% of control values 5 min after an ether stress were observed with 25 micrograms of dexamethasone, 10 mg of prednisolone and 20 mg of hydrocortisone. Although the percent inhibition of corticosterone and ACTH response to stress was comparable, a striking dissociation of the ACTH and corticosterone release was observed in terms of absolute concentrations. A mean ACTH concentration of 462 ng/l after 25 micrograms of dexamethasone was measured together with a barely measurable corticosterone concentration of 3 micrograms%. Similarly, after 10 mg of prednisolone, the mean ACTH concentration was 404 ng/l, whilst the mean corticosterone concentration was 3 micrograms%. This dissociation demonstrates that the corticosterone concentration on its own does not necessarily reflect the ACTH release. At 4 weeks of age, the ACTH response to stress is more difficult to suppress than in adult animals. This is more obvious after adrenalectomy, where the excessive ACTH secretion was less inhibited by all glucocorticoids used. The time between the last steroid gavage and stress must be considered. In 4-week-old animals the ACTH response 16 h after 12.5 micrograms of dexamethasone was inhibited by 22%, whereas 4 h after the same dexamethasone dose the inhibition was 85%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989145 TI - The relative prognostic significance of nucleolar morphometry in invasive ductal breast cancer. AB - In a group of 65 patients with invasive ductal breast cancer with adequate follow up for at least 5 years, the prognostic significance of nucleolar area morphometry was investigated. In general, non-survivors have larger and more pleomorphic nucleoli. A value of the standard deviation of the nucleolar area above 2.49 micron2 was almost exclusively found in non-survivors. This is the more interesting because several of these patients had a low mitotic activity index and small tumours, some of these having negative lymph nodes. Without nucleolar morphometry these patients would have been included in a favourable prognosis group. Although the sensitivity of nucleolar morphometry is low, the method can be used as an additional prognostic indicator. This is more important as nucleoli can in principle be automatically measured with digital image processing computers. PMID- 2989146 TI - Nuclear morphometry of epitheliosis and intraduct carcinoma of the breast. AB - Nuclear morphometry has been studied by a simple visual method in order to obtain an objective criterion for distinguishing epitheliosis from intraduct carcinoma in tissue sections. Nuclear area was found to be superior to the measurement of major or minor axes. Estimated area (major X minor axes), although less accurate than measured area, gave equally good discrimination. The best discrimination was obtained using the differences between the mean estimated nuclear area of the diseased duct and a normal duct in the same section. Using a difference of 20 micron2 as the dividing line, epitheliosis and intraduct carcinoma could be correctly classified by this criterion alone in 86% of the lesions studied. PMID- 2989147 TI - DNA hybridization in diagnostic pathology. AB - DNA hybridization is becoming an important new adjunct to conventional methods for the diagnosis of infectious diseases, inherited conditions, and neoplasia. Applications of this technology require very small quantities of tissue or body fluids because the DNA probes used in the hybridization assays detect minute amounts of homologous DNA sequence in the test material. Under the proper conditions, these DNA probes are absolutely specific for the pathogen or gene being examined, and hybridization with them usually yields objective answers that require little interpretation. The relatively minor inconveniences currently associated with DNA hybridization are related to the use of radioactivity as a detection signal and the time and labor required to obtain diagnostic data. In the future, technical improvements currently being developed and the preparation of new probes for additional human and microbial genes are likely to create an increasingly larger role for DNA hybridization in diagnostic pathology. PMID- 2989148 TI - Coxsackievirus B3 murine myocarditis: wide pathologic spectrum in genetically defined inbred strains. PMID- 2989149 TI - Elastosis in benign and malignant breast disease. AB - Elastosis, the presence of clumps of elastic fibers, is known to occur frequently in association with breast carcinoma. To test the hypothesis that the degree of elastosis increases progressively in fibrocystic disease with the severity of epitheliosis (epithelial hyperplasia, papillomatosis; widely believed to be the only premalignant component of fibrocystic disease) and increases further with intraductal and infiltrating duct carcinoma, breast tissue stained for elastic fibers from 84 women in the fifth decade of life was studied. Fourteen cases were evaluated in each of six disease categories: fibrocystic disease without epitheliosis; fibrocystic disease with epitheliosis, graded subjectively as mild, moderate, or severe (based on the degree of epithelial hyperplasia); intraductal carcinoma; and infiltrating duct carcinoma of the breast. Periductal elastosis and stromal elastosis were graded on a scale of 0 to 4 (absent to massive). The grades of both periductal elastosis and stromal elastosis were compared with those for the six disease categories ranked by increasingly advanced disease. The results indicate that the grades of periductal elastosis (Spearman rank correlation coefficient [R] = 0.54; P less than 0.001) and stromal elastosis (R = 0.75; P less than 0.001) increase progressively with the severity of breast disease. PMID- 2989150 TI - Undifferentiated (embryonal) sarcoma of the liver: ultrastructural and immunohistochemical similarities with malignant fibrous histiocytoma. AB - Two undifferentiated (embryonal) sarcomas of liver were studied ultrastructurally and immunohistochemically. Electron microscopic examination of the pleomorphic tumor cells revealed fibroblastic and histiocytic characteristics. There were no specific findings to support rhabdomyoblastic, leiomyoblastic, or epithelial differentiation. Cytoplasmic peroxidase-antiperoxidase (PAP) immunohistochemical staining for vimentin, alpha1-antitrypsin, and alpha1-antichymotrypsin was found. No staining for epidermal or internal organ cytokeratins, desmin, myoglobin, or alpha-fetoprotein was observed. The ultrastructural correlates of the cytoplasmic periodic acid-Schiff-positive, diastase-resistant hyaline globules were large, membrane-bound, heterogenous electron-dense inclusions, probably lysosomal in origin. These inclusions did not react on either alpha1-antitrypsin or alpha1 antichymotrypsin PAP staining. Tumor specimens from two metastatic sites were also examined. Neither contained the ducts or cysts that characterized the primary tumor. These studies confirm the mesenchymal nature of this uncommon childhood neoplasm and support the suggestion that the cytoplasmic hyaline globules represent a degenerative phenomenon. There are ultrastructural and immunohistochemical similarities with malignant fibrous histiocytoma. PMID- 2989151 TI - Bilateral cystic nephroblastomas and multiple malformations with trisomy 8 mosaicism. AB - The case of a 16-month-old female infant with bilateral cystic nephroblastomas, Dandy-Walker syndrome, microcephaly, bilateral cataracts, and cerebellar heterotopia is reported. The patient's older sister, who had had bilateral cystic nephroblastomas, botryoid sarcoma involving the vagina and urinary bladder, microcephaly, arhinencephaly, and bilateral cataracts, was described in a previous report. Chromosomal study in the present case confirmed trisomy 8 mosaicism (rate of mosaicism, 16 per cent). The familial occurrence and the chromosomal disorder suggest a syndrome involving genetic abnormalities. PMID- 2989152 TI - High frequency of triplicated alpha-globin loci and absence or low frequency of alpha thalassemia in Polynesian Samoans. AB - Most of the population in certain areas of Melanesia have one alpha-globin gene deletion (alpha thal2). It is thought that the high frequencies of alpha thal2 in this population is due to a selective advantage given by malaria infection to carriers of alpha thal2. We are interested in neighboring Polynesia which, although adjacent to Melanesia, has always been free of malaria due to the absence of the vector anopheles. We studied 60 Polynesian Samoans and 150 Malaysians by restriction endonuclease gene mapping using Eco RI, Bam HI, and Bgl II and hybridization to 32P-labeled alpha-globin gene probe. Seven among the 60 (11.7%) Samoans had triplicated alpha-globin loci type 1, while none had alpha thal2. On digestion with Bgl II the third alpha-globin gene was found in an additional 3.7 kb fragment in all seven Samoans with triplicated alpha-globin loci, while digestion with Bam HI produced an abnormal elongated 18.2 kb fragment carrying alpha-globin genes in addition to the normal 14.5 kb fragment. None of the Polynesian Samoans had alpha thal2 or alpha thal1. Only two of the Malaysians had triplicated alpha-globin loci. PMID- 2989153 TI - Isolation of probes detecting restriction fragment length polymorphisms from X chromosome-specific libraries: potential use for diagnosis of Duchenne muscular dystrophy. AB - We have isolated 23 human X chromosome-specific DNA fragments from lambda libraries, prepared from flow-sorted X chromosomes. To increase diagnostic potential for X-linked genetic disorders, including Duchenne muscular dystrophy (DMD), the fragments were tested for restriction fragment length polymorphisms (RFLPs) with six restriction enzymes. All fragments were regionally mapped to segments of the X chromosome with a panel of somatic cell hybrids and with human cell lines carrying unbalanced chromosomal abnormalities. Two of the isolated probes detected a high frequency RFLP. One, 754, maps between Xp11.3 and Xp21 and detects a PstI polymorphism with an allele frequency of 0.38. The other, 782, maps between Xp22.2 and Xp22.3 and reveals an EcoRI polymorphism with an allele frequency of 0.40. According to a pilot linkage study of families at risk for Duchenne muscular dystrophy, 754 gives a maximum Lod score of 7.6 at a recombination fraction of 0.03. Probe 782 lies telomeric to DMD with a maximum Lod score of 2.2 at a recombination fraction of 0.17. Using our X-chromosomal probes and a set of autosomal probes, isolated and examined in an identical way, we found a significantly lower RFLP frequency for the X chromosome as compared to the autosomes. PMID- 2989154 TI - Molecular analysis of gene deletion in aniridia--Wilms tumor association. AB - Hybrid clones were produced from the fusion of Chinese hamster cells and human fibroblasts from a patient with the aniridia-Wilms tumor association (AWTA). The DNA from the parental cells and the hybrid clones was screened by Southern blot and DNA hybridization with probes for the human insulin and Ha-ras-1 genes. Two alleles for the Ha-ras-1 gene were shown to exist in the AWTA cells by restriction fragment length polymorphism. One hybrid clone, containing a single allele for Ha-ras-1 was shown to contain a single chromosome 11 with a cytogenetically visible deletion at 11p13. The DNA from this hybrid contained the human genes for insulin, A gamma-globin, G gamma-globin, Ha-ras-1, and calcitonin, but lacked any human sequences homologous to a human catalase cDNA. This clone was also shown to express human lactate dehydrogenase A (LDH A) activity. These data indicate that the deletion of the affected chromosome in this AWTA patient begins distal to LDH A and includes band 11p13, but does not extend to calcitonin or other genes thought to be located in the distal half of chromosome 11p. PMID- 2989156 TI - Single dose intramuscular sulbactam and ampicillin in treating acute uncomplicated gonorrhoea. PMID- 2989155 TI - Immunophenotypes of lymphocytes in prospectively followed up human papillomavirus lesions of the cervix. AB - From 1981 286 women were prospectively followed up for a mean (SD) of 16 (14) months for established infection with human papillomavirus (HPV) with or without coexistent cervical intraepithelial neoplasia (CIN). The in situ immunocompetent cell infiltrates in 263 cervical punch biopsy specimens from these women were phenotypically identified by the avidinbiotin peroxidase complex (ABC) technique using monoclonal antibodies Leu-10, OKT-3, OKT-4, and OKT-8. Leu-10+ B lymphocytes far outnumbered the OKT-3+ T lymphocytes in all types of HPV lesions (flat, inverted, and papillomatous condylomas of the cervix). The ratio of OKT-4+ to OKT-8+ (T helper to T suppressor cells) was slightly reduced in HPV lesions with more severe CIN and correlated positively with the intensity of the immunocompetent cell infiltrate. The ratio of OKT-4+ to OKT-8+ cells was highest in the 47 (28.8%) patients with HPV lesions that regressed during follow up, somewhat lower in the 85 (52.1%) with persistent lesions, and lowest in the 31 (19.1%) with lesions showing clinical progression. The results are discussed in terms of the proposed immune surveillance functions attributed to immunocompetent cells in situ according to the mucosal associated lymphatic tissue (MALT) concept. The conclusion drawn is that a dynamic balance between the immunoregulatory cells and their subtypes is a prerequisite for the proper handling of intracellular infections of the mucosa, including that with HPV. PMID- 2989157 TI - 4-Aminopyridine and haemodialysis in the treatment of verapamil intoxication. AB - Verapamil, a calcium antagonist, is used as an antianginal, antidysrhythmic and antihypertensive agent. Fatal intoxications with this commonly used drug have been described. We report the effects of 4-aminopyridine and haemodialysis in a patient with severe verapamil intoxication. PMID- 2989158 TI - Cardiac involvement in coxsackie virus infection. PMID- 2989159 TI - Radionuclide assessment of left and right ventricular function following acute myocardial infarction. PMID- 2989161 TI - [Isoprenaline in the topical treatment of psoriasis]. PMID- 2989160 TI - The influence of the membrane sodium pump activity on the ability of human peripheral blood lymphocytes to form early rosettes with SRBC. AB - The influence of changes of the cation-transporting activity of the membrane Na,K ATP-ase of human peripheral blood lymphocytes on their ability to form 'early' and 'late' rosettes with SRBC was investigated. It was shown that inhibition of the Na,K ATP-ase-dependent cation transport through the lymphocyte membrane by means of incubation of the cells with various specific inhibitors of the enzyme (ouabain, vanadate) or depletion of the magnesium cations from the incubation medium caused the rise in the percentage of early rosettes, having no influence on the late E rosette percentage. The possible explanation of the observed phenomena is discussed with regard to (ion transport-dependent) electrical forces present at the surface of resting and activated lymphocytes. PMID- 2989162 TI - [Adenoma (erosive adenomatosis) of the nipple]. PMID- 2989163 TI - [Multiple trichoepithelioma, cylindroma, eccrine spiradenoma present in the same family. Histologic and histopathogenetic considerations]. PMID- 2989164 TI - DNA polymorphism of the major histocompatibility class I genes and their association with serologically defined haplotypes. AB - DNA of unrelated persons as well as members of families that were totally or partially homozygous or completely heterozygous on the loci of the major histocompatibility class I genes has been isolated from peripheral blood lymphocytes and blot hybridized with the class I pseudogene pHLA 12.4 probe. The autoradiographic DNA patterns were discussed and compared with well-defined serological features. Positive associations with serologically typed alleles had been demonstrated for HLA-A1,11; -A2; -A3; -B7; -B14; -B35; -Bw41; and -Cw5. PMID- 2989165 TI - How polymorphic are class II loci of the mouse H-2 complex? AB - DNA was isolated from 75 mouse strains carrying classical H-2 haplotypes as well as haplotypes derived from wild mice. The DNA was digested with three restriction endonucleases, Bst EII, Eco RI, and Bam HI, the digests hybridized, using the Southern blotting technique, with probes for the class II genes A alpha, A beta, E alpha, and E beta, and the restriction fragment length polymorphism at these loci determined. The analysis revealed that the most polymorphic of the four loci is A beta, followed by E beta, and, at a different level, by E alpha and A alpha. There is a large difference in the degree of polymorphism between the A beta and E beta genes, on the one hand, and the A alpha and E alpha genes, on the other hand. There is no difference in the degree of polymorphism between the A alpha and E alpha genes. These findings do not substantiate previous postulates of a high A alpha polymorphism and they do not agree with the hypothesis that the class II region is divided into highly polymorphic centromeric and less polymorphic telomeric subregions. Rather, it appears that the differences in the degree of polymorphism of the different segments of the class II region are determined by the class II loci themselves. The polymorphism of the less polymorphic class II genes is, however, still greater than the polymorphism of certain other genes on chromosome 17, notably the alpha 4-globin pseudogene. The distribution of polymorphisms at the A beta and E beta loci suggests that even populations occupying relatively small geographical regions differ in alleles at these loci. Sharing of A beta alleles between unrelated populations is yet to be detected. A certain degree of linkage disequilibrium exists among the A alpha, A beta, and E beta loci; by contrast, the E alpha locus appears to vary largely independently of the other class II loci. PMID- 2989166 TI - Functional analysis of the mouse H-2Kb gene promoter in embryonal carcinoma cells. AB - Mouse embryonal carcinoma (EC) cells do not express the major H-2 class I transplantation antigens. The latter, however, become detectable upon in vitro differentiation of EC cells. Neither class I H-2 genes nor the gene coding for beta-2 microglobulin (beta 2m) is transcribed in EC cells. We have constructed two hybrid plasmids containing the 5' flanking region of an H-2Kb gene followed by the coding regions of either the herpes simplex virus thymidine kinase (H-2 tk) or the chloramphenicol acetyl transferase (H-2 CAT) genes. Upon transfer into EC cells, the H-2 tk hybrid gene is expressed in F9 tk- cell lines which thus acquire a stable tk+ phenotype. When such transformed clones are induced to differentiate in vitro, tk activity shows a moderate increase, which reflects an increase in transcription of the hybrid gene. In transient transformation experiments, EC cells were found to express the H-2 CAT hybrid gene as well. We conclude that the 2 kilobase pair region of the H-2Kb gene which we used contains an active promoter region, but does not include all the elements required for the correct regulation of the H-2Kb gene. PMID- 2989167 TI - Mouse MHC class II gene E beta 2 is closely related to E beta and to HLA-DR beta. PMID- 2989168 TI - Anti-nuclear antibodies in murine alloantisera are not necessarily related to antibodies to murine leukemia viruses. AB - Murine alloantisera often contain antinuclear antibodies. Frequently, these sera also have antibodies to murine leukemia viruses. In order to evaluate the relationship of these activities several alloantisera were tested for the presence of antinuclear and antiviral antibodies. The results of this study show that there is neither quantitative nor qualitative correlation between antinuclear and antiviral antibodies. PMID- 2989169 TI - Effect of whole buckwheat (Fagopyrum esculentum) flour supplementation on lipid profile & glucose tolerance. PMID- 2989171 TI - The contribution of vasopressin and angiotensin to the maintenance of blood pressure after autonomic blockade. AB - The contribution of vasopressin and angiotensin II to the maintenance of blood pressure after short-term autonomic blockade was investigated in conscious Long Evans and Brattleboro (vasopressin-deficient; hereditary diabetes insipidus) rats. After short-term autonomic blockade by atropine (1 mg/kg), propranolol (5 mg/kg), and pentolinium (5 mg/kg and 10 mg/kg/hr), the fall in blood pressure was significantly greater in Brattleboro rats than in Long-Evans rats (48 +/- 3 vs 32 +/- 2 mm Hg; p less than 0.01). Administration of the vasopressin vascular receptor antagonist D(CH2)5Tyr-(Me)AVP (2 micrograms/kg) caused further blood pressure decreases only in Long-Evans rats, so that the final blood pressure in both groups was identical. Administration of enalaprilat (10 mg/kg), an angiotensin converting enzyme inhibitor, further reduced blood pressure in both strains. When enalaprilat was given first after autonomic blockade, it reduced blood pressure in Brattleboro rats but not in Long-Evans rats. Administration of the vasopressin antagonist after enalaprilat further reduced blood pressure only in Long-Evans rats. The fall in blood pressure following vasopressin blockade was greater than that occurring after angiotensin converting enzyme inhibition (14 +/ 1 vs 6 +/- 1 mm Hg; p less than 0.05) in autonomic blockade Long-Evans rats. Plasma levels of vasopressin in Long-Evans rats increased markedly after short term autonomic blockade, whereas plasma renin and angiotensin II levels were unchanged. Plasma angiotensin II levels were increased by the vasopressin antagonist and decreased by enalaprilat. We conclude that, due to sympathetic nervous system blockade and consequent blunting of renal renin release, vasopressin has a greater capacity than the renin-angiotensin system for maintaining blood pressure after short-term autonomic blockade. PMID- 2989170 TI - Atriopeptins as cardiac hormones. PMID- 2989172 TI - Membrane endopeptidases of human neutrophil. AB - Recent studies using inhibitors or synthetic substrates of serine protease suggest that membrane protease activity may be essential for neutrophil chemotaxis, phagocytosis, degranulation, and superoxide production. However, little is known about the nature and localization of the proteases. In this study, we demonstrated that intact human neutrophils hydrolyzed [125I]glucagon. The degradation of glucagon was temperature dependent and was not dependent on the release of lysosomal enzymes. Two endopeptidases were demonstrated: a metalloendopeptidase which accounted for two thirds of the intact cell activity, and a serine endopeptidase, accounting for the rest of the activity. Both enzymes had a neutral to alkaline pH optimum (pH 7-9). The metalloendopeptidase had a Km of 15.3 microM and Vmax of 5.9 nmol/5 X 10(6) cells/45 min. The corresponding values for the serine endopeptidase were 33.3 microM and 5.0 nmol/5 X 10(6) cells/45 min, respectively. Inhibition of the membrane metalloendopeptidase or serine endopeptidase by 1,10-phenanthroline or diisopropylfluorophosphate, respectively, did not inhibit the production of superoxide by phorbol myristate acetate-stimulated neutrophils. PMID- 2989174 TI - Increased release of hydrogen peroxide and superoxide anion from asbestos-primed macrophages. Effect of hydrogen peroxide on the functional activity of alpha 1 protease inhibitor. AB - The ability of asbestos-elicited murine peritoneal macrophages to release superoxide anion and hydrogen peroxide, following in vitro triggering, has been investigated. The asbestos-elicited macrophages produced increased levels of superoxide and hydrogen peroxide compared to control macrophages and similar levels to those produced by Corynebacterium parvum elicited macrophages. The supernatants from asbestos-elicited macrophages which had been triggered in vitro were capable of impairing the ability of alpha 1-protease inhibitor to inhibit elastase function. The catalase sensitivity of this effect showed it to be due to hydrogen peroxide. PMID- 2989173 TI - Characterization of effect of N-formyl-methionyl-leucyl-phenylalanine on leukotriene synthesis in human polymorphonuclear leukocytes. AB - Human polymorphonuclear leukocytes (PMNLs) were exposed to N-formyl-methionyl leucyl-phenylalanine (f-Met-Leu-Phe) in the presence or absence of exogenous arachidonic acid. Analysis of incubation mixtures by high-performance liquid chromatography showed that f-Met-Leu-Phe stimulated the synthesis of 5-hydroxy eicosatetraenoic acid (HETE) and of leukotriene B4 (LTB4) which was rapidly metabolized into 20-OH-LTB4 and 20-COOH-LTB4. The stimulatory effect of f-Met-Leu Phe was dose and time dependent. The tripeptide showed maximum stimulatory activity at the concentration of 1 microM and after 20-30 min of incubation. Addition of arachidonic acid to the f-Met-Leu-Phe-stimulated PMNLs resulted in an increase in the synthesis of LTB4 and 5-HETE. Pretreatment of the PMNLs with cytochalasin B strongly potentiated (up to six-fold) the stimulatory effect of f Met-Leu-Phe on 5-lipoxygenase product synthesis, whereas cytochalasin B alone or with arachidonic acid had no significant effect. The tripeptide did not increase the synthesis of platelet-derived 12-HETE, and 12-hydroxyheptadecatrienoic acid, or of PMNL-derived 15-HETE, suggesting that its action was selective for PMNL 5 lipoxygenase. The present data indicate that f-Met-Leu-Phe causes the release of arachidonic acid from cellular lipids and activates the 5-lipoxygenase. PMID- 2989176 TI - Role of stimulated neutrophils from patients with systemic lupus erythematosus in tissue injury, with special reference to serum factors and increased active oxygen species generated by neutrophils. AB - To examine the possible correlation between tissue injury and neutrophil-produced active oxygen (AO) species in patients with systemic lupus erythematosus (SLE), we studied the capacity of the serum from six patients with untreated, active SLE to generate AO and release lysosomal enzymes by normal neutrophils. Cultured endothelial cells from human umbilical cord vein were incubated with serum stimulated neutrophils to assess AO-induced tissue injury. Serum from patients with bacterial infections and healthy individuals served as controls. AO production was highest in the neutrophils stimulated with SLE patient-derived serum, while lysosomal enzyme release was only slightly increased. SLE neutrophils with or without stimulation and SLE serum-stimulated normal neutrophils produced significantly high levels of cytotoxicity upon coincubation with 51Cr-labeled human endothelial cells. These excessive cytotoxicities were reversed by the presence of superoxide dismutase and catalase, indicating the specificity of the AO effect on endothelial cell damage. These findings suggest that tissue damage in SLE may be partially due to excessive production of AO and that both neutrophils themselves and a serum factor which activates neutrophils are involved in the mechanism for vascular injury. PMID- 2989175 TI - Effect of diethyldithiocarbamate (DDC) on human monocyte function and metabolism. AB - Recent evidence indicates that phagocytic cells play a major role in tissue inflammation. The release of enzymes, lipid metabolites such as prostaglandins, and reactive oxygen species by these cells appear to mediate the inflammatory process. In this study we have evaluated the effects of diethyldithiocarbamate (DDC) on human monocyte function and metabolism. We demonstrate that DDC impairs that antibody-dependent cytoxicity (ADCC) of monocytes to red cell targets. The concentration of DDC which caused maximal suppression of ADCC also prevented the burst of oxidative metabolism in monocytes stimulated by sensitized red cells targets or phorbol myristate acetate (PMA). DDC also impairs the lipid metabolism of these cells as indicated by a decrement in malonyldialdehyde (MDA) production. These data indicate that DDC impairs the activity of two major biochemical pathways in monocytes which are related to the inflammatory process, i.e., the release of oxygen metabolites and prostaglandins. PMID- 2989177 TI - Human monoclonal antibodies against Mycobacterium leprae. AB - Human hybridomas were constructed which produce antibodies against three different extracts of Mycobacterium leprae. A thioguanine-resistant (Thgr), ouabain-resistant (Ouar), human lymphoblastoid cell line, KR-4, was hybridized with Epstein-Barr virus-transformed cell lines from lepromatous leprosy patients with fusion frequencies of greater than 10(-5). Non-Epstein-Barr virus transformed donor cells fused at much lower rates (less than 2 X 10(-7]. Hybrids were selected in medium containing hypoxanthine aminopterin thymidine and 10(-5) M ouabain. An enzyme-linked immunosorbent assay was used to screen for antibodies against three crude extracts of armadillo-derived M. leprae, including (i) a soluble sonic extract preparation, (ii) sodium dodecyl sulfate extract of insoluble sonicated M. leprae, and (iii) a purified phenolic glycolipid antigen. Of a total of 2,200 final clones screened, 359 were found to secrete antibody which bound to soluble sonic extracts and the sodium dodecyl sulfate extract (6.7 and 9.6%, respectively), whereas 12.5% (21 out of 168) showed positivity to the glycolipid antigen. Four selected hybridomas also reacted with the deacylated derivative of M. leprae phenolic-glycolipid antigen. The specificity of these monoclonal antibodies was partially determined by screening on a panel of crude extracts from four other mycobacteria. Nine clones of 122 showed reactivity to M. leprae only. The predominant immunoglobulin was immunoglobulin M, and quantities up to 10 micrograms/ml were produced. Antibody production by hybrid clones was stable in more than 75% of the clones grown in continuous culture. By comparison, 10,000 Epstein-Barr virus-transformed lymphocyte clones from lepromatous leprosy patients were screened for anti-M. leprae antibody production, and all of the 42 clones that were initially positive in the enzyme-linked immunosorbent assay lost their antibody-producing capabilities within 6 weeks in culture. These results suggest that a combination of Epstein-Barr virus transformation and hybridization may be an optimal method in producing human monoclonal antibodies from leprosy patients. PMID- 2989178 TI - Susceptibility of Propionibacterium acnes to killing and degradation by human neutrophils and monocytes in vitro. AB - Propionibacterium acnes, the target of inflammation in acne, was tested for its sensitivity to the bactericidal and degradative functions of human polymorphonuclear leukocytes (PMN), monocytes, and their fractions. P. acnes strains were not killed by PMN under any conditions and were variably killed by monocytes in the presence of serum from acne patients. Control strains of Staphylococcus aureus and Micrococcus lysodeicticus were susceptible to both PMN and monocyte killing. P. acnes strains were also not killed by lysozyme, chymotrypsin, H2O2, human serum, PMN granule lysate, and PMN and monocyte cell lysates. The organism was sensitive to the bactericidal activity of myeloperoxidase in acid pH. In addition, P. acnes was shown to be relatively resistant to the degradative action of PMN and monocyte lysates, whereas M. lysodeicticus, S. aureus, and Staphylococcus epidermidis were all degraded to various degrees. The moieties that were liberated from P. acnes by PMN enzymes were predominantly low in molecular weight (1,000 to 25,000) and were consistent with cell wall fragments. PMID- 2989179 TI - Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. AB - A large plasmid is found in virulent isolates of Shigella sp. and encodes functions essential for invasion of mammalian cells. To identify plasmid sequences necessary for invasion, we isolated a series of Tn5 insertions in pWR100, the virulence plasmid of Shigella flexneri serotype 5. These insertions demonstrated that three separate EcoRI fragments of pWR100 were required for invasion of HeLa cells. However, the corresponding native EcoRI fragments, when cloned into pBR325, did not restore virulence to plasmidless strains. Construction of a lambda-sensitive, plasmidless Shigella recipient enabled us to shotgun clone plasmid DNA directly into S. flexneri by using the cosmid vector pJB8 and score for expression of invasive functions. In this fashion, we succeeded in isolating six independent recombinants which restored invasion of HeLa cells in plasmidless Shigella recipients. The cloned inserts all contained a common core of ca. 37 kilobases, thus defining a minimum sequence necessary for invasion of HeLa cells. Virulence-associated peptides produced by wild-type S. flexneri were also produced by the recombinants. Expression of these peptides and expression of invasiveness by the clones were regulated by growth temperature, as is expression of these traits in wild-type S. flexneri. A complete invasive phenotype was not expressed by the recombinants in that they failed to produce a positive Sereny test. Possible explanations for this behavior as it relates to the mechanism of bacterial invasion are discussed. PMID- 2989180 TI - Effect of estrogen (17 beta-estradiol) on the susceptibility of mice to disseminated gonococcal infection. AB - Studies of the effect of sex hormones on the susceptibility of mice to the disseminated gonococcal infection demonstrated significantly enhanced susceptibility of mice injected with estrogen (17 beta-estradiol). In mice treated with estradiol, bacteremia progressively developed within 12 h postinoculation and mice died within the next 6 h, whereas bacteremia in mice treated with progesterone was completely cleared within 3 h postinoculation. The administration of estradiol affected the function of polymorphonuclear leukocytes (PMN) responsible for eliminating gonococci, but the administration of progesterone did not. The bactericidal activity of PMN mediated by myeloperoxidase was affected by estradiol, but the capacity of PMN to release superoxide anion was not. Furthermore, peritoneal cell analysis demonstrated that the infiltration of PMN in the peritoneal cavity of estradiol-treated mice significantly decreased when mice were injected intraperitoneally with gonococci. These effects on PMN by estradiol may play an important role in the enhanced susceptibility of estradiol-treated mice to gonococcal infection. PMID- 2989181 TI - Enhanced superoxide release and tumoricidal activity by a postlavage, in situ pulmonary macrophage population in response to activation by Mycobacterium bovis BCG exposure. AB - The monocytic phagocyte population of rat lungs is heterogeneous. In addition to the freely lavagable alveolar macrophages, there is a fixed in situ tissue associated subpopulation of pulmonary macrophages. The response of this subpopulation to classical macrophage activation by Mycobacterium bovis BCG exposure was monitored. Results indicate that this population can be activated both metabolically and functionally, as evidenced by enhanced release of superoxide anions and demonstrable tumoricidal activity against syngeneic and xenogeneic target cells. The pattern of metabolic activation of in situ tissue associated macrophages differed somewhat from that of alveolar macrophages and was observed only after subsequent exposure of the cells to either zymosan particles or phorbol myristate acetate. Upon such exposure, the activated zymosan treated tissue macrophages released approximately twice as much superoxide as the nonactivated cells and amounts comparable to the amounts released by activated alveolar macrophages. The tissue macrophages also displayed greater levels of cytotoxicity toward xenogenic targets than the alveolar cells and may have an important role in preventing microbial or tumor cell colonization of respiratory systems. PMID- 2989182 TI - Mechanism of polymyxin B-mediated lysis of lipopolysaccharide-treated erythrocytes. AB - A novel system was used previously to characterize the dynamic interaction of a polysaccharide-deficient, lipid-rich lipopolysaccharide (LPS) with rabbit erythrocytes (RaRBC). Exposure of the RaRBC to the LPS rendered them sensitive to induction of hemolysis by the cationic antibiotic polymyxin B (PB) in a time- and temperature-independent manner. Subsequent decay in the response of LPS sensitized cells to PB was shown to be critically dependent on both the time and temperature of incubation of RaRBC with LPS and to be independent of a change in LPS binding (Carr and Morrison, Infect. Immun. 43:600-606, 1984). In the present study, we performed experiments designed to define the mechanism by which PB mediates hemolysis of LPS-sensitized RaRBC. Experiments were performed to examine the molecular requirements of the LPS and the PB that were essential for hemolytic activity. The capacity of various cations to mediate hemolysis of LPS sensitized RaRBC or to block PB-mediated hemolysis and the temperature dependence of the PB lytic reaction were investigated. The results of these experiments suggest that PB-mediated hemolysis of LPS-treated erythrocytes is dependent upon an initial ionic association of PB with erythrocyte membrane-bound LPS, followed by hydrophobic insertion of the PB fatty acid into the erythrocyte membrane lipid bilayer. PMID- 2989183 TI - Vascular integrity and platelet function. PMID- 2989184 TI - New functions for platelets and their pathological implications. AB - We have recently demonstrated in Schistosoma mansoni infection that rat and human platelets could very efficiently kill parasite larvae, both in vivo and in vitro. The study of this IgE-dependent platelet effector function has led us to several subsequent findings. They concern: (1) the existence of a specific receptor for IgE on the platelet surface; (2) its close association with a platelet membrane glycoprotein of essential functional importance, the GPIIb-IIIa complex; (3) the observation, in extrinsic allergic asthma, of an allergen-specific IgE-dependent platelet activation; (4) the identification, in aspirin-sensitive asthma, of a similar, but non-IgE-dependent, platelet activation selectively induced by cyclo oxygenase inhibitors, and prevented by salicylate. Beyond their implication in anti-parasite immunity, these findings provide a basis for new insights on the participation of platelets in disease. PMID- 2989186 TI - The acquired immunodeficiency syndrome (AIDS): an update. AB - The acquired immunodeficiency syndrome (AIDS) is a disease which is characterized by a profound defect in cell-mediated immunity leading to opportunistic infections and unusual neoplasms such as Kaposi's sarcoma. It is caused by a retrovirus of the human T cell leukemia/lymphoma virus family and has been termed HTLV-III/lymphadenopathy-associated virus. The virus selectively infects the T4 helper/inducer subset of T cells, accounting for the profound defects in immunity noted in AIDS patients. AIDS is transmitted sexually, and by blood and blood products, accounting for its relative confinement to specific risk groups. The disease is now seen worldwide, and as of September 1984, approximately 6,000 cases have been reported in the USA alone. There is no effective treatment for the disease at present, and the mortality of the full-blown syndrome approaches 100%. Hopefully, with the recent isolation of the causative agent of the syndrome, effective therapies and vaccinations will be forthcoming. PMID- 2989185 TI - Adenosine receptors on human inflammatory cells. AB - Adenosine is a natural nucleoside that plays a physiological role in the modulation of human inflammatory cells. We have investigated the presence of adenosine A2/Ra and A1/Ri receptors and of the P-site on human inflammatory cells. Human B and T(OKT4+ and OKT8+) lymphocytes, polymorphonuclear leucocytes, monocytes, basophils and platelets possess a membrane adenosine A2/Ra receptor. The activation of adenosine A2/Ra receptor increases the intracellular level of cyclic AMP in these cells. Human lymphocytes and neutrophils possess also an inhibitory adenosine A1/Ri receptor and a P-site whose activation inhibits the effect of many adenylate cyclase agonists including isoproterenol, PGE1, histamine, adenosine, cholera toxin and forskolin. PMID- 2989187 TI - Administration of adenosine diphosphate-ribosyl transferase antagonist allows in vivo control of anti-dinitrophenyl response. AB - 3-Methoxybenzamide (3MB) is one of a series of chemical inhibitors of the nuclear enzyme adenosine diphosphate (ADP)-ribosyl transferase (ADPRT), which has been shown to inhibit cell differentiation in vitro, but has no effect on differentiation independent proliferation. Treatment of mice with an optimal concentration of 3MB (20 mg/kg body weight) at or 1 day after dinitrophenyl keyhole limpet haemocyanin (DNP-KLH) immunisation reduced anti-DNP plaque-forming cell (PFC) numbers to less than 10% of those of control animals. The period for maximum PFC suppression showed a narrow time window relative to immunisation, suggesting that in vivo, as in vitro, 3 MB was acting only on those lymphocytes differentiating in response to antigen. Experimental findings showed that it was possible to select for PFC derived from different populations of DNP-responsive lymphocytes by adjusting the time of 3MB treatment relative to immunisation. When 3MB was used with antigen priming, the residual PFC showed a lower average affinity than PFC in mice treated with 3MB 3 days after priming, suggesting a differential selection of those lymphocytes responding either 'early' or 'late' in the primary immune response. PMID- 2989188 TI - Decreased calcium dependence of lymphoblastoid cell lines compared with Burkitt lymphoma cell lines. AB - The extracellular calcium level required for proliferation was compared in B lymphoid cell lines from various sources by determining the calcium concentration at which long-term proliferation was inhibited by 50% (CaPD50). Fourteen Burkitt lymphoma (BL) lines had a mean CaPD50 of 44 +/- 28 microM whereas 45 lymphoblastoid cell lines (LCLs) obtained by in vitro transformation of B lymphocytes with Epstein-Barr virus (EBV) had a mean CaPD50 of 3.6 +/- 1.8 microM. This difference applied also to autologous BL lines and LCLs established from the same patient. The decreased calcium requirement of virally-transformed compared with tumour-derived cell lines therefore appears to be a universal phenomenon in mammalian cells. Within the BL group, no correlation was found between the calcium requirement for proliferation and presence or absence of the EBV genome. Arrest of BL lines and LCLs occurred in the G1 phase of the cell cycle and was readily reversed by addition of calcium to the medium. One anomalous LCL was found which showed a high CaPD50 (43 +/- 6 microM) and accumulated in both G1 and G2. These results, in combination with a previous study of EBV transformation in vitro, indicate that the calcium dependence of B lymphocytes generally decreases in the following order: normal cells greater than BL cells = early stage transformation greater than LCL. The 2 transformed phenotypes thus distinguished in human lymphoid cells may offer unique opportunities for defining the status and expression of EBV in vitro and in vivo. PMID- 2989189 TI - Incidence of adult T-cell leukemia-lymphoma and its familial clustering. AB - The yearly incidence of ATLL in the Uwajima district is 6.6 patients per 100,000 inhabitants aged over 40. The yearly morbidity rate from ATLL of persons in this district who are positive for HTLV-antibody and older than 40 is 1 patient per 1,631. Familial occurrence was observed in 9/38 families available for pedigree analyses. Even in the endemic area, the existence of positive HTLV antibody is remarkably high in ATLL families, suggesting that HTLV has been transmitted from generation to generation mainly within these particular families. PMID- 2989190 TI - Mononuclear cells infiltrating human mammary carcinomas: immunohistochemical analysis with monoclonal antibodies. AB - Breast carcinomas were examined by the immunoperoxidase technique using antisera specific for lymphocyte subsets, monocytes, NK cells and major histocompatibility antigens (HLA-A, -B, -C; Ia-like). Sixty-four per cent of the patients had a moderate or strong mononuclear cell infiltration, 77% of the patients without mononuclear cell infiltration had receptors for estrogens as compared to 51% of the patients with infiltration. The majority of the infiltrating mononuclear cells were T cells; generally the OKT8 cells were predominant. The Leu 3A/OKT8 cell ratio was not related to histological type, tumor size, age of the patient or presence of metastases. Some of the T cells had the Ia antigen and were thus probably activated. The B cells were either absent or less numerous than the T cells. There was no relation between their distribution and the various parameters studied. A few monocytes were heterogeneous according to their markers (OKM I and acid phosphatase). In 6 cases only there was a strong infiltration of mononuclear cells positive for acid phosphatase. The number of the natural killer cells was also low. Only a few mononuclear infiltrating cells had receptors for transferrin. There was a positive correlation between the inflammatory infiltration and the presence of HLA class-I antigens on tumor cells. Some of the antisera specific for lymphocyte subsets also stained the breast carcinoma cells. The great variations in the subsets of mononuclear cells in breast carcinomas may correspond to various systems of defense against neoplasm. PMID- 2989191 TI - Regional variation in prevalence of antibody against human T-lymphotropic virus types I and III in Kenya, East Africa. AB - The prevalence of antibodies against HTLV-III and -I was studied among populations of 6 distinctly different regions of Kenya, an equatorial African country in which AIDS has rarely been observed. Overall, 21% of subjects had ELISA reactions suggesting the presence of antibody against HTLV-III. The frequency of HTLV-III antibodies was highest among the Turkana people (50%) and lowest among the Masai (8%). Prevalence increased with age but was not related to sex. The pattern of ELISA-detected antibody against HTLV-I was similar. The specificity of these antibodies was supported by Western blot analysis of a subset of sera with high and low ELISA ratios, in which 66% and 73% of those with ELISA ratios considered positive (= greater than 5.0 in this study) also had a profile of bands consistent with HTLV-III and HTLV-I respectively. The antibodies detected were not cross-reactive between HTLV-III and HTLV-I on Western blot analysis. In a series of subjects with various parasitic and infectious diseases, patients with idiopathic splenomegaly and with schistosomiasis had a high proportion of antibodies against both HTLV-III and HTLV-I. This survey shows that reactivity in the ELISA HTLV-III and HTLV-I assays are common among Kenyans but vary considerably by region. PMID- 2989192 TI - Monoclonal-antibody-defined human lung tumour cell-surface antigens. AB - A panel of 3 monoclonal antibodies (MAbs) directed against human lung tumour cell surface antigens has been produced following immunizations with the established small-cell lung cancer (SCLC) cell line, NCI-H69, and with another SCLC cell line, COR-L32, recently derived from clinical material. One MAb, B10/12, reacted strongly with SCLC, immunoprecipitated a protein having an MW of 100kd and failed to react significantly with non-small-cell lung cancer (NSCLC) in radioimmunoassay and in an immunohistochemical assay. MAbs E10/5 and 2G3 reacted extensively with SCLC but also showed significant reactivity with NSCLC. MAb E10/5 immunoprecipitated a protein with an MW of 80kd but no appreciable protein was specifically precipitated by MAb 2G3. Unlike MAb 2G3, both MAbs B10/12 and E10/5 reacted strongly with selected neuroblastomas whereas only MAbs 2G3 and E10/5 reacted significantly with melanoma. All 3 MAbs reacted with breast carcinomas. Other non-pulmonary tumours thus far examined failed to react with the MAbs in radioimmunoassay or immunohistochemical assay. Immunocytochemistry and the use of viable cells in radioimmunoassay confirmed that the antigenic determinants recognized by these MAbs were surface located. PMID- 2989193 TI - Leukotrienes. PMID- 2989194 TI - Congenital polyonychias. Reduction versus duplication digit malformations. PMID- 2989195 TI - Poly(ADP-ribose) and the recovery from damage in Chinese hamster cells due to 5 bromodeoxyuridine photolysis. AB - Exposure of Chinese hamster cells to near-u.v. light, following the uniform incorporation of 5-bromodeoxyuridine (BrdUrd) into their DNA, resulted in cell killing that was close to exponential. An inhibitor of poly(ADP-ribose) synthesis, 3-aminobenzamide (3-ABA), enhanced the cytotoxic effect of this treatment when present for 2 h at 20 mM after light exposure. The dose modifying factor was 1.4. Under conditions that resulted in a sigmoidal survival curve (a 30 min BrdUrd pulse in S phase, followed 90 min later by light exposure) the effect of 3-ABA was to remove the shoulder of the survival curve with very little change in its final slope. Using various inhibitors of ADP-ribosyl transferase (ADPRT) the enhanced cell killing was found to correlate with the inhibitors' relative potency. Cellular NAD+, the substrate for poly(ADP-ribose) synthesis, was rapidly depleted after exposure. This depletion was largely prevented by 3 ABA; the activity of ADPRT increased with the fluence of near-u.v. light; and the concentration of cellular NAD+ decreased with exposure. ADPRT activity was maximal immediately after exposure to near u.v. light and then decayed to pre exposure levels within 30 min (37 degrees C). The enhanced cytotoxicity of BrdUrd + near-u.v. light, when followed by 3-ABA treatment, disappeared at a rate similar to that of the decay in ADPRT activity. We conclude from these results that poly(ADP-ribose) synthesis is important for the recovery from BrdUrd photolysis damage in DNA. Because this damage and its repair are relatively specific (e.g. compared to ionizing radiation) and relatively easy to manipulate, it could serve as a model system for the study of the role of poly(ADP-ribose) in the repair of DNA damage. PMID- 2989196 TI - Enzymatic restriction of mammalian cell DNA: evidence for double-strand breaks as potentially lethal lesions. AB - Permeabilized Chinese hamster cells were treated with the restriction endonucleases Pvu II and Bam H1 which generate blunt-ended and cohesive-ended DNA double-strand breaks (dsb), respectively. Cells were then assayed for their clonogenic ability. These experiments were performed to test the hypothesis that mammalian cell death following X-ray exposure arises from the induction of dsb in DNA, and via the formation of chromosomal aberrations. It was shown previously that Pvu II induces chromosome aberrations whereas Bam H1 was ineffective in this respect. The results reported here show that Pvu II simulates X-ray exposure, in causing a dose-dependent loss of the reproductive integrity of mammalian cells. Dsb generated by Pvu II, i.e. with blunt ends, can therefore be regarded as potentially clastogenic as well as potentially lethal. Bam H1 was found not to reduce cell survival in the same enzyme dose range. These results support the notion that X-irradiated mammalian cells undergo a mode of death in which dsb in the DNA cause chromosomal aberrations which are lethal as a result of loss of genetic material in the form of chromosome fragments, or as a result of chromosome bridge formation. PMID- 2989197 TI - The labeling of high affinity sites of antibodies with 99mTc. AB - Labeling of F(ab')2 with 99mTc was investigated. The best labeling procedure for F(ab')2 was then applied to IgG and Fab. Stannous ion was used as the reducing agent for 99mTcO-4 and free DTPA was used as a competing reagent to prevent colloid formation and loosely bound 99mTc. The competition reactions revealed two 99mTc binding sites for F(ab')2 and IgG. One is a high capacity, low affinity site. This accounts for 76 and 84% of total IgG and F(ab')2 binding sites. The labeling of these sites can be prevented if the antibody is labeled in the presence of free DTPA. The second site is a low capacity, high affinity site. The labeling of these sites cannot be prevented by free DTPA. Fab, unlike IgG and F(ab')2, does not have an appreciable percentage of high affinity sites. The determination of sulfhydryl groups using Ellman's reagent indicates the production of 5.5, 4.2 and 0.9 sulfhydryl groups when IgG, F(ab')2 and Fab were exposed to 56 micrograms/mL SnCl2 X 2H2O. These sulfhydryl groups may be the source of the high affinity binding. Biodistribution in mice for 99mTc labeled F(ab')2 and F(ab')2-DTPA, both prepared in the presence of excess free DTPA, was similar to that of F(ab')2-DTPA-111In. PMID- 2989198 TI - Assessment of right ventricular ejection fraction during intravenous 133Xe pulmonary ventilation study. AB - Intravenous 133Xe in saline was used to estimate right-ventricular ejection fraction (RVEF). In three healthy volunteers, the first pass RVEF with 133Xe was followed by RVEF estimation by intravenous 99mTc-pertechnetate. There was agreement between the 133Xe and 99mTc determined values. Subjects rebreathed the liberated 133Xe gas, allowing an estimation of regional pulmonary ventilation. Intravenous 133Xe in saline may have potential use in measuring RVEF, as well as in evaluating pulmonary function in cor pulmonale. PMID- 2989199 TI - Management of inflammation: lipid peroxide as a parameter for a better understanding of inflammatory processes. AB - The management of inflammation, based on an overall, coordinated observation of the inflammatory system, requires the assessment of the validity and biological significance of the various components in inflammation, a procedure that is more reliable than the interpretation of individual measurements in isolation. Such an overall approach, here developed from the model of carrageenin-induced inflammation, also predicates that lysosomal enzymes, lipid peroxide and proamidase (related, respectively, to the inflammatory response in a narrow sense, to tissue damage and to tissue repair) are three basic parameters required when studying inflammatory processes. This overall approach, moreover, shows that the inflammatory process must be regarded as being diachronic, and that in the light of the observation of exudate fluid it can be divided into three main stages, namely an early stage of exudation related to lipid peroxide, an intermediate stage of exudate dilution related to granuloma amidase, and a later stage of exudate concentration related to granuloma amidase and plasma protein. PMID- 2989200 TI - Management of inflammation: proamidase as a parameter for a better understanding of inflammatory processes. AB - Coordinated observation of various factors in the inflammatory system and their relations shows that the levels of proamidase in granuloma pouch fluid in carrageenin-induced inflammation increase during the healing process whereas the proamidase shows (1) remarkably low values in rats injected with high doses of carrageenin, (2) low values in the earlier stages of inflammation with tissue damage (high values of fluid lipid peroxide and lactate dehydrogenase) and with phagocytosis (high values of fluid peroxidase and beta-glucuronidase), and (3) high values in the later stages where wound healing is progressing (high values of protein in plasma, fluid and granuloma pouch tissue and low values of fluid volume, fluid peroxidase, beta-glucuronidase and pouch tissue weight). In short, proamidase shows low values in the stage of tissue damage and constantly increasing values during the process of healing. Thus the measurement of proamidase, which reflects the degree of wound healing, provides an important parameter for a broad, coordinated observation of the inflammatory process, and may enable the management of inflammation. PMID- 2989202 TI - Ocular manifestations of the acquired immune deficiency syndrome. PMID- 2989201 TI - Report of a WHO meeting on AIDS. PMID- 2989203 TI - Inflammation and hypersensitivity. AB - The leukotrienes are a family of potent mediators of hypersensitivity and inflammation. The application of sensitive and specific radioimmunoassays for leukotrienes has revealed the generation of significant quantities of several leukotrienes by leukocytes exposed to natural stimuli in the absence of exogenous arachidonic acid. The C6 peptide leukotrienes, LTC4 and LTD4, are potent vasoactive and smooth muscle contractile factors. LTB4 and other di-HETEs stimulate polymorphonuclear leukocyte function and suppress T-lymphocyte activities. The effects of LTB4 on T lymphocytes are subset-specific and include the activation of suppressor and cytotoxic T cells. Receptors for LTB4 on polymorphonuclear leukocytes are specific but exhibit considerable heterogeneity of binding affinity and may mediate different functional effects. The complexity of the pathways of generation and metabolism of leukotrienes, and of the leukotriene receptors, suggest that carefully defined systems will be needed to test the effects of pharmacological inhibitors and antagonists. PMID- 2989204 TI - Mechanisms of DNA reorganization in bacteria. PMID- 2989205 TI - Transposable elements in eukaryotes. PMID- 2989206 TI - [Corticotropin releasing factor (CRF): diagnostic aspects]. PMID- 2989207 TI - [Neurosurgical therapy of neuroendocrinologic disorders]. PMID- 2989208 TI - [Psychoneuroendocrinology]. PMID- 2989209 TI - Quantitation of herpes simplex virus in rabbit corneal epithelium. AB - The authors have developed an objective method for quantitation of herpes simplex virus in the corneal epithelium of rabbits. At appropriate times postinfection, full-thickness rabbit corneas were removed by trephination and subjected to one cycle of freezing and thawing. The corneal epithelium was then disrupted by sonication. The amount of infectious virus recovered from sonicated specimens was determined by an in vitro plaque assay, providing a measure of the quantity of virus present during the acute stage of herpetic keratitis. Using this technique, the authors found that the mean virus titer was reduced from 1.5 X 10(5) plaque forming units (pfu) per cornea in control rabbits to less than 200 pfu per cornea in rabbits treated topically for 2 days with 1% trifluridine. In contrast, instillation of 1% prednisolone acetate resulted in the persistence of higher levels of virus (275 pfu) than those observed in control rabbits (3 pfu) 4 days after the cessation of therapy. PMID- 2989210 TI - The relationship between tonic accommodation and ciliary muscle innervation. AB - Previous studies have used Badal laser optometry to demonstrate significant variance in the distribution of tonic (or "dark-focus") resting positions of accommodation (TA) for groups of observers. This study investigates whether individual differences in TA are due to individual differences in autonomic tone of the ciliary muscle by comparing separately the effects of a muscarinic receptor antagonist (Tropicamide 0.5%) and a beta receptor agonist (Isoprenaline 3%) on the distribution of TA. Ten subjects were used for each study with mean ages of 23.2 and 23.0 yr, respectively. The distribution of TA within each subject group was equivalent to the large sample distributions published in previous literature. Darkroom measurements of TA and pupil diameter were determined with a Badal laser optometer and infrared photography. Distance correction and amplitude of accommodation were determined by standard optometric techniques. Isoprenaline produced a significant hyperopic shift in TA of 0.47 D over 22 min but this was not accompanied by a significant change in standard deviation of the TA distribution. This finding was in accord with the authors' previously reported investigation using the beta receptor antagonist Timolol Maleate. Tropicamide also produced a significant hyperopic shift in TA of 1.24 D over 24 min, and this was accompanied by a significant change in standard deviation of the distribution from 0.87 to 0.17. The results indicate that the parasympathetic system plays a significant role in determining the TA position and that the variations in TA between individuals is a consequence of parasympathetic rather than sympathetic ciliary muscle tone. PMID- 2989211 TI - Distribution of calpain I, calpain II, and calpastatin in bovine lens. AB - Two types of Ca2+-requiring cysteine proteinase (calpain, EC 3.4.22.17), which required for full activation 100 microM Ca2+ (calpain I) and 1 mM Ca2+ (calpain II) were found to exist in the cytosolic fraction of bovine lens. Since calpain may play an important role on the degradation of lens proteins during the aging process of the lens, we attempted to study the distribution of calpain I, calpain II, and calpastatin (an endogenous specific inhibitor of calpain) in bovine lens. It was found that both the capsule-epithelium and cortex fiber cells contained calpains I and II and calpastatin, although the content of calpain I was much lower than that of calpain II. Calpains I and II and calpastatin activities were not detected in the nuclear region at all. PMID- 2989212 TI - Synaptic lamellae of the photoreceptors of pearl and wild-type mice. AB - The retina of the pearl mutant mouse, C57BL/6J pe/pe, exhibits reduced light sensitivity in the dark-adapted condition (Balkema and Pinto, J Neurophysiol 48:968, 1982). The authors searched for an anatomic correlate in the retina which could relate to the functional deficit. Electron microscopic mosaics of the outer plexiform layer of light- and dark-adapted pearl and wild-type mice were analyzed. The numerical density and length of the photoreceptor synaptic lamellae showed these parameters to be indistinguishable in wild-type and pearl retinas under conditions of both light- and dark-adaptation. Light-adapted pearl retinas exhibited some rod spherules that contained structurally modified synaptic lamellae with bulbous thickenings and adjacent electron-dense bodies. These lamellar modifications were neither apparent in the light-adapted, wild-type retinas, nor in the dark-adapted retinas of either genotype. Pronase application to ultrathin sections hydrolyzed synaptic lamellae, bulbous thickenings and electron dense bodies. PMID- 2989213 TI - Herpes simplex virus type 1 and neuronal cells--a special cell-virus interaction. AB - Rat brain glioma cells were semipermissive for herpes simplex virus (HSV) replication, because the growth of HSV was multiplicity-dependent in these cells. By using this property, we successfully isolated 'survivor' glioma cells following HSV infection at low multiplicity and without using any special treatment (such as UV irradiation) either of the cells or of the virus. Under the same conditions there were no survivor BHK or 3T3 cells, which suggests the uniqueness of the glioma cell-HSV interaction. The survivor cells ceased to produce infectious virus after two subcultures, but were highly resistant to superinfection for at least 20 subcultures. Parental cells were significantly more permissive for homologous virus growth than survivor cells. Interferon was apparently not induced in the survivor cells, because they were as susceptible as the parental cells to infection with vesicular stomatitis virus. The survivor cells produced HSV-specific antigens and contained HSV-specific DNA. PMID- 2989214 TI - Congenital guinea pig infection with attenuated Junin virus strains. AB - Guinea pigs born from mothers infected before or during pregnancy with 10(3) PFU of the attenuated XJC13 or XJ0 strains of Junin virus (JV) by the intramuscular route showed 31.5% mortality that was not attributable to the mothers' clinical condition or to lack of care. There was a slight drop in mortality rate when the mothers were infected at the beginning or end of their gestation period. JV isolation from the 9 offspring killed from 1 to 125 days of age proved that virus transmitted transplacentally or soon after birth was able to persist, although titers were not higher than 10(2.7) PFU/g of tissue in various organs, including brain. Cell-associated viremia could thus account for viral spread after birth. Since an active humoral response was detected in the same animals, although Nt antibody titers were below 1:16, a state of tolerance did not exist in these congenitally infected animals. The carrier state appeared to modify guinea pig susceptibility to JV; after challenge with the pathogenic XJ strain of JV, 2 animals survived and developed normal humoral responses, while half of the remaining animals did not show typical signs of Argentine hemorrhagic fever. Although JV persistence appeared to cause no deleterious effects in surviving guinea pigs, its long-term risk remains to be determined. PMID- 2989215 TI - Cirrhosis and hepatocellular carcinoma: a 13 year necropsy study with attention to the role of alcohol. PMID- 2989216 TI - Nephrectomy in childhood--a ten-year review. PMID- 2989217 TI - Horse heart acylphosphatase: purification and characterization. AB - The use of an affinity chromatography step performed with an immunoadsorbent consisting of anti-horse muscle acylphosphatase antibodies covalently linked to Sepharose 4B allowed us to purify horse heart acylphosphatase in a very rapid and efficient fashion. As in skeletal muscle, also in heart the enzyme is present as both a mixed disulfide with glutathione and a S-S dimer. The abundance of these forms in heart is quite lower than in skeletal muscle. The comparison of the molecular forms so purified with those obtained from horse skeletal muscle showed the same aminoacid composition, tryptic fingerprint, together with strictly similar apparent molecular weight and main kinetic parameters, supporting the conclusion that the acylphosphatase present in heart is the same enzyme as that purified from skeletal muscle. PMID- 2989218 TI - [Leukotrienes and monohydroxy fatty acids: controversial role in the pathogenesis of psoriasis]. AB - The pros and cons concerning the involvement of arachidonic acid metabolism in the pathogenesis of psoriasis are presented. The isolation of arachidonic acid metabolites from psoriatic lesions, their extraordinary biological activity, and the therapeutic efficiency in psoriasis of inhibitors of arachidonic acid metabolism all argue in favor of leukotrienes and monohydroxy fatty acids playing an important role in the development of psoriasis plaques. On the other hand, the lack of specificity of the biochemical findings, the failure to reproduce psoriatic lesions by arachidonic acid metabolites, and the therapeutic activity of drugs that have no effect on arachidonic acid metabolism show that the role of arachidonic acid metabolism in the pathogenesis of psoriasis is still controversial. The availability of selective inhibitors of arachidonic acid metabolizing enzymes for clinical testing is a prerequisite before pathophysiological conclusions can be made, as the present status of knowledge makes any conclusions premature. PMID- 2989219 TI - [The importance of adrenergic receptors for dermatology]. AB - Receptor research is a rather new but most promising field of research. This review summarizes the data available on the adrenergic receptors. in addition, information is provided about the important function of these membrane receptors as control or modulator of dermatophysiology and dermatopathy. PMID- 2989220 TI - [Hereditary sclerodactyly and syndactyly]. AB - A 61-year-old man is described with sclerodactyly of the hands and syndactyly of the second and third toes. Hereditary sclerodactyly is a rare condition, beginning in early youth with flexion contracture of the fingers. In this patient the skin of the fingers was sclerotic and thickened, and the dorsal skin of the hands was atrophic and dry. The condition did not progress nor did it show signs of Raynaud's phenomenon. Both feet showed syndactyly of the second and third toes. The family tree suggested autosomal dominant inheritance, with reduced penetrance since the grandfather of our patient was reported to have had a similar disease. PMID- 2989221 TI - Events leading to the treatment of coronary artery disease: implications for nursing care. AB - The selection of a particular treatment for CAD was found to be related to the severity of illness, patients who were more severely disabled by angina elected surgical treatment after vessel blockage was documented by angiography. Differential access to treatments was not described by study subjects and therefore cannot be evaluated as a contributing factor to the selection of treatment. Although most subjects participated in a therapeutic pharmacologic regimen, few patients in either group employed dietary or exercise programs to ameliorate CAD. Most patients received information about CAD and about surgery as a treatment option. This educational communication was directed by the cardiologist for a large majority of patients. Nurses were cited as information sources for surgical subjects only. In retrospect, more than half of the surgical subjects indicated a need for additional information. The decision regarding which type of treatment to have was made autonomously by most subjects, unlike the usual decision-making styles of families of surgical patients. A significantly greater proportion of surgical patients identified their family as a source of support during the elected treatment. Both groups described family changes specific to the illness and selected treatment. PMID- 2989222 TI - Patterns of enzyme activities in the gills of the catfish Heteropneustes fossilis (Bloch) exposed to the anionactive detergent Na-alkyl-benzenesulphonate (LAS). AB - The effect of sodium-alkyl-benzenesulphonate (LAS) on the activity of the respiratory enzymes of the gills of Heteropneustes fossilis (Bloch) was investigated. After 48 h exposure, the main injury to gills was the progressive separation of the lamellae from their vascular components. The enzymes of the aerobic part of the metabolism showed a decrease in activity, whereas the activity of lactate dehydrogenase was strongly increased, thus indicating that LAS has a high potential to interfere with aerobic mechanisms; however, the mode of action of it has yet to be clearly defined. PMID- 2989224 TI - [Quantitative functional scintigraphy of the salivary glands--clinical possibilities and current indications]. AB - A special background subtraction in dynamic scintigraphy of the salivary glands enables the production of net-time-activity-curves and the calculation of quantitative parameters of each gland. Regional functional differences within a single gland are shown by phase and gradient images. The physiological kinetics of the tracer and its modifications in different diseases of the glands (acute and chronic inflammation, tumours, Sjogrens-Syndrome) are analyzed in 367 patients. PMID- 2989223 TI - Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals. AB - The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not detectable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase. PMID- 2989225 TI - Southern analysis of DNA polymorphism among Dw subtypes of DR4. AB - DNA from individuals of four Dw subtypes of DR4 (Dw4, Dw10, Dw14, Dw15) were studied using Southern blotting to determine if subtype-specific DR beta or DQ beta restriction fragment polymorphism could be found. Although very little polymorphism was found among ten DR4 homozygous individuals (4 Dw4, 2 Dw10, 3 Dw14, 1 Dw15) using a Dr beta or a DQ beta probe, restriction fragment polymorphism was easily detected between different DR types (DR1-DRw8). The possible evolutionary significance of the lack of Dw-associated polymorphism relative to DR-associated polymorphism is discussed. PMID- 2989226 TI - Interferon production by mitogen-stimulated human lymphocytes requires differentiation but not DNA synthesis. AB - Mitogens such as phytohemagglutinin (PHA) stimulate T lymphocytes to differentiate, proliferate, and produce lymphokines: e.g., interferon gamma (IFN gamma) and interleukin-2 (IL-2). Induction of IFN gamma is an early event in mitogenesis, accompanied by an increase in RNA and protein synthesis, and later followed by DNA synthesis. Although, in general, good IFN gamma inducers are good mitogens, we show here that DNA synthesis is not an obligatory requirement for the production of IFN gamma. Aphidicolin and hydroxyurea, which completely inhibit DNA synthesis, do not significantly affect IFN production, whether added before, with, or after mitogen stimulation. Inhibitors of differentiation, however, affect DNA and IFN synthesis depending on their time of addition in relation to PHA. The inhibitors, 3-methoxybenzamide (3MB) and a group of compounds that affect cyclic nucleotide metabolism (theophylline, aminophylline, and dibutyryl cyclic AMP), had profound inhibitory effects on IFN production. However, bromodeoxyuridine (BUdR), also an inhibitor of differentiation in some systems, had very little effect on IFN production. PMID- 2989227 TI - The proliferative immune response to autologous Epstein-Barr virus-transformed lymphoblastoid cells. I. Studies with HLA haplotype loss variants demonstrate a role for MHC-linked genes. AB - The human Epstein-Barr virus transformed lymphoblastoid cell line (EBV-LCL) 721 and MHC haplotype loss variants derived from it were utilized to dissect the functional role of MHC genes in the proliferative response of autologous T lymphocytes to EBV-LCL. LCL-721 is heterozygous at all phenotypically defined MHC loci. One type of LCL-721 variant expresses only determinants encoded by the maternal (m) haplotype and the other expresses determinants encoded by the paternal (p) haplotype. Autologous (individual A) primary proliferative responses are strong to each type of haplotype deletant. The strong tertiary responses to the priming haplotype in comparison to the relatively weak responses to the reciprocal haplotype indicate that MHC linked genes encoded by each haplotype are important in the autologous response to EBV-LCL. Similar specific tertiary responses are observed when peripheral blood lymphocytes (PBLs) from the donor's mother are used as responding cells. Allogeneic responses were also studied by priming PBLs from unrelated donors with the haplotype deletants. Quantitative comparisons of the proliferation by primed allogeneic and autologous lymphocytes stimulated by irradiated PBLs from donor A and her mother, and by LCL-721 and its variants, show that some of the tertiary responses involve specific recognition of EBV-LCL while others detect recognition of alloantigens. PMID- 2989229 TI - Breast relapse following primary radiation therapy for early breast cancer. II. Detection, pathologic features and prognostic significance. AB - In order to define better the nature of breast relapse following primary radiation therapy for early-stage invasive breast cancer, we examined the clinical patterns, methods of detection, histopathology and prognosis in 25 patients. Eighty-eight percent of these relapses occurred in the vicinity of the original tumor at an average of 33 months after treatment. Twelve percent occurred in a location distinctly separate from the primary tumor area at an average of 75 months after treatment. In 14 patients breast relapse was detected by physical examination (PE) alone, in 6 patients by mammography alone and in 5 patients by both. In eight of the patients whose relapses were detected by PE alone, mammography was not performed; mammograms were negative in the other six. In 89% of the patients who presented with a new abnormality on physical examination, the recurrence consisted predominantly or exclusively of invasive carcinoma. In contrast, in all six patients who presented with only a new mammographic abnormality, the recurrence consisted predominantly or exclusively of intraductal carcinoma. Eighteen of these patients underwent salvage mastectomy, 3 were inoperable on clinical grounds and 4 refused mastectomy. Only 1 of the 18 patients who underwent mastectomy had a significant complication related to the surgery. Twenty-one of these 25 patients (84%) are alive without further recurrence up to 67 months (mean 24 months) after breast relapse. We conclude that (1) physical examination and mammography are both important for the detection of breast relapses: (2) secondary (salvage) surgery can be performed without significant complications; and (3) a breast relapse does not have the same grave prognosis as a local (chest wall) recurrence after mastectomy. PMID- 2989228 TI - In vitro development of human monoclonal antibody-secreting plasmacytomas. AB - A polyclonal human lymphoblastoid cell line transformed in vitro with Epstein Barr virus produced specific anti-Rhesus D antibody. It was repeatedly enriched by rosetting procedures and subsequently cloned. The cloning conditions employed a combination of mouse macrophage feeder layers, antimycoplasma agents, and low density passage. Formal evidence of monoclonality was obtained in one case which was of human IgG1 isotype and was secreted at the level of 15-20 micrograms/ml. All clones showed long-term stability in culture after 10 months of continuous passage. Both polyclonal and monoclonal cell lines possessed antigens characteristic of highly differentiated B cells, yet they also expressed Epstein Barr virus nuclear antigen (EBNA). This study exemplifies a simple method for obtaining monoclonal antibody secreting plasmacytomas of human origin. PMID- 2989230 TI - Pediatric brain stem tumors: patterns of treatment failure and their implications for radiotherapy. AB - There have been conflicting opinions regarding the correct volume to be used in radiotherapy fields for brain stem tumors of childhood. Whereas many clinicians recommend limited fields designed to cover the tumor volume with a margin, some have advocated whole brain radiotherapy. Using our clinical experience at Duke University Medical Center, we have made an attempt to determine the proper irradiation volume in this group of tumors. We have evaluated 38 patients with brain stem tumors in children less than 18 years of age. The most common presenting symptoms were headache, ataxia, and hemiparesis. Thirteen patients had a histologic diagnosis made prior to treatment or post-mortem. All had either an anaplastic astrocytoma or a glioblastoma multiforme. Tumors were located in the thalamus, hypothalamus, or midbrain in 9 patients and in the pons or medulla oblongata in the remaining 29 patients. All patients received a course of radiotherapy. The mean minimum tumor dose was 52.6 +/- 5 Gy given at 1.7 to 2.0 Gy/fraction. Twenty-three patients received radiation to a limited field and 14 received whole brain irradiation. In one patient, the field size could not be ascertained. The five year survival of the total group was 39%. The survival of patients with thalamic, hypothalamic, or midbrain tumors was 73% compared with 28% for those with tumors of the pons or medulla oblongata (p = 0.0159). Eighty eight percent of the tumor recurrences in evaluable patients (22/25) occurred within the radiotherapy fields. Patients were stratified for tumor location and no difference was observed in survival or relapse-free survival among those individuals treated with limited irradiation fields or whole brain irradiation fields. When our results are examined in conjunction with previously published data, the bulk of existing evidence supports the use of limited fields for irradiation of brain stem tumors of childhood. PMID- 2989231 TI - Prognostic factors in patients undergoing curative irradiation for breast cancer. AB - We have evaluated 147 patients who received primary curative irradiation for breast cancer to determine if there were any common factors that predisposed to failure. There were 10 local recurrences, for an overall local control rate of 93.2%, with a 2 to 16 year follow-up. Disease-free survivals were 85% in Stage I, 69% in Stage II, and 50% in Stage III. Age seemed to be a determinant, with a 31.6% local recurrence rate in women under 50 vs. a 3.1% rate of local recurrence in women age 50 or older when treated. We have also noted a correlation between the radiation dose and local recurrence, with a recurrence rate of 9.1% following a dose of 6000 rad or less, vs. a rate of 4.7% with a dose of 6400 rad or more. The size of the primary carcinoma and the status of the axillary lymph nodes, each considered individually, were not significant prognostic factors. There was no relationship between recurrence and the performance of an axillary dissection. There was evidence suggesting that the recurrence rate was lower when a wide local excision was performed, compared to a lesser procedure. Multiple foci of carcinoma and a high nuclear grade appear to have a poor prognosis. Five of the recurrences occurred early (under 2 years) and three of these were accompanied by fatal distant metastases. None of the five patients with late recurrences (37 to 116 months) developed metastatic disease, suggesting that late recurrences may have a better prognosis. PMID- 2989232 TI - CT scanning for radiation therapy treatment planning of hepatoma. AB - One hundred forty-five patients with hepatoma had CT scanning for radiation therapy treatment planning. In order to demonstrate the anatomical distortions that occur with hepatoma and its effect on treatment planning, a control group of 50 colorectal cancer patients with normal livers was analyzed for comparison. The objectives of planning were to deliver as homogeneous a dose to the whole liver as possible and not to treat more than one of two functional kidneys or more than one-half of both functional kidneys. Conventional AP/PA portals were defined by physical examination, intravenous pyelogram, and bowel gas patterns at simulation and were found to be inadequate for the treatment of 76% of patients with hepatoma and 10% of patients with normal livers. Among the control group patients with no hepatoma, only 10% required oblique portals and 6% could not be treated because of left hydronephrosis or a solitary right kidney. Because the distortion of the liver in hepatoma in relationship to the kidneys required portal modification in 76% of hepatoma cases; 39% required oblique planning, 24% AP/PA, 20% PA and left lateral portals, and 17% required 4-field, 3-field or other plans in order to meet the treatment planning objectives. We concluded that all patients receiving radiation therapy to the liver for hepatoma require CT scanning for optimum radiation therapy treatment planning because of the hepatic distortion that occurs in hepatoma and the requirements of renal tolerance. PMID- 2989233 TI - Lactation following primary radiation therapy for carcinoma of the breast. PMID- 2989234 TI - Studies on antiviral agents. I. Synthesis and in vitro antiviral activity of new kanamycin A derivatives. PMID- 2989235 TI - Excitatory amino acid pharmacology of the auditory nerve and nucleus magnocellularis of the chicken. AB - Experiments were performed on the nucleus magnocellularis and auditory nerve in tissue slices of 19-20-day-old chick embryos. Bath-applied kainate, quisqualate and N-methyl-D-aspartate induced dose-dependent alterations in the antidromic responses of nucleus magnocellularis neurons. The sensitivity of these agonist induced responses to 2,3-cis-piperidine dicarboxylate, glutamate diethylester and D-alpha-aminoadipate were tested, as was the sensitivity of auditory nerve transmission. The data suggest that receptors for all three agonists are present on nucleus magnocellularis neurons and that the postsynaptic receptor of the nucleus magnocellularis-auditory nerve synapse is of the kainate type. The effects of bath-applied baclofen were also studied. Baclofen blocked orthodromic responses suggesting that an excitatory amino acid is released from the presynaptic terminal. PMID- 2989236 TI - Evaluation of the use of deuterium oxide dilution techniques for determination of body composition of beef steers. AB - Body composition as estimated by a one- or two-compartment deuterium oxide dilution technique was compared with directly measured body composition of 15 large- and 15 small-frame steers. Body composition of the steers was measured at 219, 412 and 603 kg live weight. Empty body protein was overestimated (P less than .05) 3.6% from a one-compartment model (1 CM, using the slope, intercept method), while empty body protein was underestimated (P less than .05) 5.4% from a two-compartment kinetic model (2CM). Empty body ether extract estimated by 1 CM was not significantly different from the direct method, although 4.7% larger. Empty body ether extract was overestimated (P less than .001) 32.2% by the 2CM. Empty body water was accurately estimated from the 1CM when a 3.2% correction factor was used for the overestimation of total body water by the 1CM, but water in gastrointestinal tract contents was overestimated (P less than .001) 13.4% by the 1CM. Empty body water was underestimated (P less than .001) 7.8% by the 2CM, and water in gastrointestinal tract contents was overestimated (P less than .001) 41.8% by the 2CM due to its dependence on regression equations that differ between groups of cattle. The 2CM offered no advantage over the 1CM. A three compartment model was not better than the 2CM in estimating body water compartments. Assuming the amount of empty body water associated with either empty body protein or ash to be constant seemed to be valid. Suggested values calculated from data presented in the literature for growing cattle with an empty body weight greater than 175 kg are .302 and .0668, respectively, for the ratios of protein and ash to water. The relationship between empty body fat and water was, percentage empty body fat = 94.27--(1.267)(percentage empty body water), which had a 1.25 residual standard deviation and a .98 coefficient of determination. PMID- 2989237 TI - Effects of dietary alfalfa:orchardgrass hay and lasalocid on nutrient utilization by gravid sows. AB - Twenty-four third- and fourth-parity crossbred gravid sows were housed in metabolism stalls and randomly allotted to four dietary treatments: corn alfalfa:orchardgrass hay (46%; CH), CH plus lasalocid (CHL), corn-soybean meal (CS) or CS plus lasalocid (CSL). Daily feed intake was 2.36 kg for CH- and CHL fed sows and 1.66 kg for CS- and CSL-fed sows. Lasalocid intake was 110 mg X sow 1 X d-1. Total feces and urine were collected during two 5-d collection periods, 40 and 80 d postcoitum. Digestible energy (DE), metabolizable energy (ME), nitrogen (N) retention, dry matter (DM), neutral detergent fiber (NDF) and acid detergent fiber (ADF) digestibilities, and retention of calcium (Ca), total phosphorus (P), magnesium (Mg), sodium (Na) and potassium (K) were determined. Percentage DE and ME were greater (P less than .0001) for CS- and CSL-fed sows (88.3 and 83.4%) than for CH- and CHL-fed sows (65.2 and 61.3%). Lasalocid did not influence (P greater than .05) DE and ME values but decreased (P less than .01) the ME:DE ratio. Percentage N digestibility was greater (P less than .0001) for CS- and CSL-fed sows (85.7%) than for CH- and CHL-fed sows (60.8%), but there was no significant difference in N retention, expressed as a percentage of N intake. Lasalocid increased (P less than .03) percentage N digestibility; however, the diet X additive interaction decreased (P less than .005) grams N retained X sow-1 X d-1 for CSL-fed sows. The CH- and CHL-fed sows digested more grams of DM, NDF and ADF but had lower percentage digestibilities of DM and fiber fractions than did CS- and CSL-fed sows. The diet X additive interaction decreased (P less than .05) percentages of Ca, P and K digested and decreased (P less than .05) grams mineral retained (Ca, P, Mg and K) and mineral retained expressed as a percentage of intake for CSL-fed sows. Results indicate that alfalfa:orchardgrass hay and lasalocid influenced energy, fiber, nitrogen and mineral utilization by gravid sows. PMID- 2989238 TI - Site and extent of starch and neutral detergent fiber digestion as affected by source of calcium and level of corn. AB - Five steers (385 kg) fitted with permanent abomasal cannulae were used to compare Ca source (limestone or dicalcium phosphate) and corn level on site and extent of digestion. Diets contained 50, 70 or 90% corn, with corn silage and supplement to provide .70% Ca from either limestone or dicalcium phosphate. Limestone did not affect ruminal digestion, but postruminal starch and neutral detergent fiber (NDF) digestion were higher (P less than .05) for limestone compared with dicalcium phosphate, which suggests that starch utilization may be increased postruminally by a compound with buffering capacity. As a percentage of total starch intake, total tract and ruminal starch digestion increased (linear, P less than .01) while postruminal starch digestion decreased (linear, P less than .01) with corn level. Neutral detergent fiber digested in the rumen decreased (linear, P less than .01) and postruminal NDF digestion increased (linear, P less than .01) when level of corn in the diet increased. Effects of corn level were not different when organic matter and starch intake were included as covariates. PMID- 2989239 TI - Multiplicity dependence and sensitivity of herpes simplex virus isolates to antiviral compounds. AB - An immunoassay that enables one to assess both viral multiplicity and sensitivity to antiviral drugs, was used to determine the sensitivity of untreated patients' virus isolates. The upper border levels for judging clinical isolates sensitive to Ara-A, ACV, PFA or IDU were calculated from a total of 48 primary herpes simplex isolates. Although five isolates were considered less sensitive to one drug and one isolate to two drugs, all were sensitive at a lower virus multiplicity. Analysis of these isolates showed that no isolate was genetically resistant, but that multiplicity dependence of drugs was high with ACV and Ara-A, lower with PFA. In view of the multiplicity dependence of HSV sensitivity to different drugs, it is recommended that isolates from treatment failures or isolates considered resistant should be assayed in detail. PMID- 2989240 TI - Role of microtubules in surfactant secretion. AB - In the isolated perfused rat lung and cultured type II cells, surfactant secretion and cellular adenosine 3',5'-cyclic monophosphate (cAMP) content was stimulated by beta-adrenergic agonists. Isoproterenol-induced surfactant secretion was inhibited by the antimicrotubule agents colchicine and vinblastine. Incorporation of [3H]glycerol into disaturated phosphatidylcholine was augmented by beta-adrenergic agents but was not significantly different from the enhanced incorporation rate when colchicine was present. This suggests that the augmented incorporation of [3H]glycerol into disaturated phosphatidylcholine was a secondary response to storage depletion rather than direct cAMP stimulation. beta Adrenergic agents shifted the equilibrium in the isolated perfused rat lung and cultured type II cells to favor microtubules. The stimulatory effect of 1.0 microM isoproterenol on tubulin polymerization was observed as early as 1 min and was augmented 2.8-fold at a half-maximal stimulation of 4 nM in cultured type II cells. Cytochalasin B, an antimicrofilament agent, potentiated the isoproterenol induced secretion. These results suggest that an intact microtubule-microfilament system may be obligatory for enhanced surfactant secretion and that beta adrenergic agents not only induce surfactant release but also tubulin polymerization. PMID- 2989241 TI - Development of beta-adrenergic control of phospholipid secretion in rabbit lung. AB - Lung distension is associated with increased phospholipid secretion into the air spaces. Basal, lavage-induced, and inflation-produced phospholipid secretion, in postmortem in situ lungs of newborn rabbits, were examined at three different levels of maturity, with and without 10(-3) M dl-propranolol. Lungs were lavaged with saline at successive 3- and 15-min time intervals to separate basal from lavage-induced secretion. Inflation-produced secretion was studied after static inflation at 30 cmH2O for 30 min. At 27.5 days gestation, basal secretion was undetectable, and neither lavage-induced nor inflation-produced secretion were influenced by propranolol. At 29.5 days gestation, basal secretion was only just detectable. Distension-associated secretion was increased over that present at 27.5 days gestation, and propranolol had a significant inhibitory effect, especially on lavage-induced secretion, in which the inhibition was shown to be rapidly reversible. There was a significant increase of basal secretion at 2.5 days postterm, possibly inhibited by propranolol. In addition, there was a further substantial increase of distension-associated secretion, and the inhibitory effect of propranolol persisted. These changes were independent of the sedimentation behavior of lavaged phospholipid. Overall, the results are consistent with evidence, produced in other laboratories, that there is an increasing density of sympathetic neurons and beta-adrenergic receptors in whole lung preparations during late gestation in the rabbit and suggest that granular pneumocytes, the presumed source of secreted phospholipid, take part in this developmental change. PMID- 2989242 TI - Production of an auto-stimulatory growth factor by human hepatoma cells abrogates requirement for a brain-derived factor. AB - Human hepatoma cells grow at high cell density in the absence of exogenous growth factors. At low cell density, two different hepatoma cell lines required a novel growth factor from brain tissue. A factor with similar physico-chemical properties in the concentrated medium from high density cultures completely substituted for the brain extract. The autogenous secretion of a novel liver cell growth factor that is concentrated in brain tissue may underlie in part the unregulated growth of hepatomas. PMID- 2989243 TI - Free radical damage to cultured porcine aortic endothelial cells and lung fibroblasts: modulation by culture conditions. AB - Culture conditions modulating cell damage from xanthine plus xanthine oxidase derived partially reduced oxygen species were studied. Porcine thoracic aorta endothelial cells and porcine lung fibroblasts were maintained in monolayer culture. Cells were prelabeled with 51Cr before xanthine plus xanthine oxidase exposure. Endothelial cells showed 30 to 100% more lysis than fibroblasts and thus seemed more sensitive to this oxidant stress. The effect of cell culture age, as indicated by population doubling level (PDL), was examined. Response of low PDL endothelial cells and fibroblasts subjected to oxidant stress was compared with the response of PDL 15 cells. Both low PDL endothelial cells and fibroblasts responded differently to the lytic effect of xanthine oxidase-derived free radicals than did higher PDL cells. Specific activities of the antioxidant enzymes catalase, manganese superoxide dismutase, copper-zinc superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase were measured in both low and high PDL fibroblasts and endothelial cells. Antioxidant enzyme specific activities could only partially explain the differences in response to oxidant stress between fibroblasts and endothelial cells and between low and high PDL cells. Cell culture medium composition modulated the rate of production, and relative proportions of xanthine plus xanthine oxidase-derived partially reduced species of oxygen, i.e. superoxide, hydrogen peroxide, and hydroxyl radical. Serum content of medium was important in modulating free radical generation; superoxide production rates decreased 32%, H2O2 became undetectable, and hydroxyl radical generation decreased 54% in the presence of 10% serum. The medium protein and iron content also modulated free radical generation. The data suggest that cell culture media constituents, cell type, and cell culture age greatly affect in vitro response of cells subjected to oxidant stress. PMID- 2989244 TI - Slow-growing Rhizobium japonicum comprises two highly divergent symbiotic types. AB - We examined the interrelationships of the genomes of 10 slow-growing strains of Rhizobium japonicum to provide a foundation for molecular genetic studies of these agriculturally important endosymbiotic bacteria of commercial soybeans. The degree of base substitution in and around known symbiotic genes (nif and presumptive nod), constitutively expressed genes (glnA and recA), and two other cloned sequences was estimated from restriction site variation by using cloned DNAs as hybridization probes to genomic Southern blots. Two highly divergent patterns of conservation of nifDH genes and nod-homologous sequences were found. On this basis, we classified the strains as the symbiotic genotypes sTI or sTII. Existing maps of the nif genes of R. japonicum apply only to strains of the sTI genotype. This division was further characterized by four other probes which also distinguished two sublines within sTI. Phenograms were constructed depicting interrelationships according to DNA sequence divergence. sTI and sTII are two highly divergent evolutionary lines consistent with the status of individual species. Neither is related to fast-growing Rhizobium strains (PRC strains) nodulating soybeans. PMID- 2989245 TI - Direct cloning of the trxB gene that encodes thioredoxin reductase. AB - A strain was constructed which contains mutations in the genes encoding thioredoxin (trxA) and thioredoxin reductase (trxB) such that filamentous phage f1 cannot grow. The complementation of either mutation with its wild-type allele permits phage growth. We used this strain to select f1 phage which contain a cloned trxB gene. The location of the gene on the cloned fragment was determined, and its protein product was identified. Plasmid subclones that contain this gene overproduce thioredoxin reductase. PMID- 2989246 TI - Molecular cloning of the structural gene coding for CTP:CMP-3-deoxy-manno octulosonate cytidylyltransferase from Escherichia coli K-12. AB - The kdsB gene from Escherichia coli K-12, which encodes CTP:CMP-3-deoxy-manno octulosonate cytidylyltransferase (CMP-KDO synthetase), was cloned into pBR322 as an 8-kilobase PstI fragment. Selection of this cloned segment was facilitated by using Salmonella typhimurium SL5283, which is deficient in three restriction enzyme systems and thus allows efficient cloning of E. coli DNA in S. typhimurium. The temperature-sensitive kdsB gene from S. typhimurium HD2 was transduced into strain SL5283 after the insertion of transposon Tn10 near the kdsB allele. Tetracycline-sensitive variants of strain SL5283 were then derived and used to select clones of the E. coli K-12 gene, inserted into the PstI site of pBR322, by complementation of the temperature-sensitive lesion in kdsB. One plasmid, pRG-1, complemented the kdsB temperature-sensitive allele and had the following characteristics: (i) it coded for several polypeptides by coupled transcription-translation in vitro, including one polypeptide which comigrated with CMP-KDO synthetase during polyacrylamide gel electrophoresis in sodium dodecyl sulfate; (ii) it overproduced CMP-KDO synthetase activity 20- to 40-fold depending on strain and growth conditions; and (iii) it coded for activity of CMP KDO synthetase which, when purified to homogeneity, had the same molecular weight and kinetic characteristics as CMP-KDO synthetase of chromosomal origin. PMID- 2989247 TI - Cloning and mapping of the genetic determinants for microcin B17 production and immunity. AB - Plasmid pMccB17 (70 kilobases [kb]) codes for the production of microcin B17, a peptide that inhibits DNA synthesis, and for microcin B17 immunity. A BamHI-EcoRI fragment of 5.1 kb from pMccB17 was cloned into pBR322 in two steps. The resulting plasmid (pMM102) overproduced microcin B17 and expressed immunity against microcin. Mcc- and Mcc- Imm- mutants were isolated on plasmids pMccB17 and pMM102 by deleting various DNA fragments and by inserting different translocatable elements. Physical and phenotypic characterization of these mutants showed that a DNA region of 3.0 to 3.5 kb is required to produce microcin B17, whereas an adjacent region of about 1.0 kb is required to express microcin B17 immunity. PMID- 2989248 TI - Genes aroA and serC of Salmonella typhimurium constitute an operon. AB - Genetic analysis of aroA554::Tn10 derivatives of two mouse-virulent Salmonella typhimurium strains, "FIRN" and "WRAY," and of a nonreverting derivative of each constructed for use as a live vaccine, showed the site of the insertion among mapped aroA point mutants. The WRAY live-vaccine strain gave no aro+ recombinants in crosses with aroA point mutations to one side of the insertion, indicating a deletion from Tn10 through the sites of these point mutations. The FIRN live vaccine strain gave wild-type recombinants with all tested point mutants; it probably has a deletion or inversion extending from Tn10 into aroA but not as far as the nearest point mutation. Some tetracycline-sensitive mutants of aroA554::Tn10 strains required serine and pyridoxine, indicating loss of serC function, and some that were found to be SerC- did not produce gas from glucose, indicating a loss of pfl function. These results show the gene order pfl-serC aroA, as in Escherichia coli. Ampicillin enrichment applied to pools of tetracycline-sensitive mutants of strains with Tn10 insertions near aroA (i.e., zbj::Tn10 strains) yielded Aro- SerC- Pfl-, Aro- SerC+ Pfl+, and Aro- SerC- Pfl+ mutants but none which were Aro+ SerC-. All of the mutants are explicable by deletions or inversions extending clockwise from zbj::Tn10 into or through an operon comprising serC (promoter-proximal) and aroA. Such an operon was also shown by the identification of two Tn10 insertions causing phenotype Aro- SerC-, each able to revert to Aro+ SerC+ by precise excision. serC corresponds to the open reading frame promoter-proximal to aroA that was identified elsewhere by base sequencing of a cloned aroA segment of S. typhimurium (Comai et al., Science 221:370-371, 1983). Both serine and chorismate are precursors of enterochelin; this may be why serC and aroA are in a single operon. PMID- 2989249 TI - Proton translocation coupled to dimethyl sulfoxide reduction in anaerobically grown Escherichia coli HB101. AB - Proton translocation coupled to dimethyl sulfoxide (DMSO) reduction was examined in Escherichia coli HB101 grown anaerobically on glycerol and DMSO. Rapid acidification of the medium was observed when an anaerobic suspension of cells, preincubated with glycerol, was pulsed with DMSO, methionine sulfoxide, nitrate, or trimethylamine N-oxide. The DMSO-induced acidification was sensitive to the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (60 microM) and was inhibited by the quinone analog 2-n-heptyl-4-hydroxy-quinoline-N-oxide (5.6 microM). Neither sodium azide nor potassium cyanide inhibited the DMSO response. An apparent----H+/2e- ratio of 2.9 was obtained for DMSO reduction with glycerol as the reductant. Formate and H2(g), but not lactate, could serve as alternate electron donors for DMSO reduction. Cells grown anaerobically on glycerol and fumarate displayed a similar response to pulses of DMSO, methionine sulfoxide, nitrate, and trimethylamine N-oxide with either glycerol or H2(g) as the electron donor. However, fumarate pulses did not result in acidification of the suspension medium. Proton translocation coupled to DMSO reduction was also demonstrated in membrane vesicles by fluorescence quenching. The addition of DMSO to hydrogen saturated everted membrane vesicles resulted in a carbonyl cyanide p trifluoromethoxyphenyl-hydrazone-sensitive fluorescence quenching of quinacrine dihydrochloride. The data indicate that reduction of DMSO by E. coli is catalyzed by an anaerobic electron transport chain, resulting in the formation of a proton motive force. PMID- 2989250 TI - Expression of symbiotic genes of Rhizobium japonicum USDA 191 in other rhizobia. AB - A 200-megadalton plasmid was mobilized from Rhizobium japonicum USDA 191 to other Rhizobium strains either that cannot nodulate soybeans or that form Fix- nodules on certain cultivars. The symbiotic properties of the transconjugants indicate that both soybean specificity for nodulation and cultivar specificity for nitrogen fixation are plasmid encoded. PMID- 2989251 TI - Amiloride-sensitive Na+-H+ antiporter in Escherichia coli. AB - In everted vesicles of Escherichia coli, delta pH caused by H+ efflux through the Na+/H+ antiporter was measured by using a fluorescent dye. Amiloride inhibited the activity of the Na+/H+ antiporter. Kinetic studies showed that amiloride competed with Na+. The inhibition constant of 40 microM was obtained. PMID- 2989252 TI - Cloning of the contiguous 165-kilobase-pair region around the terminus of Escherichia coli K-12 DNA replication. AB - Escherichia coli K-12 chromosomal DNA was partially digested with either EcoRI or HindIII, and cosmid libraries were constructed. By screening these libraries, a series of partially overlapping clones which covered the terC region was isolated. The cloned area spanned about 165 kilobase pairs, corresponding to the 29.7-to-33.2-min region of the genetic map of the E. coli chromosome. PMID- 2989253 TI - Organization of the Tn6-related kanamycin resistance transposon Tn2680 carrying two copies of IS26 and an IS903 variant, IS903. B. AB - The kanamycin resistance transposon Tn2680, which originates from the R plasmid Rts1, is homologous to Tn6 and carries two directly repeated copies of IS26, one at each end. The kanamycin resistance gene codes for type I aminoglycoside-3' phosphotransferase. Tn2680 also contains, in the middle of the transposon, an additional IS element homologous to IS903. This element, designated IS903.B, is flanked by a 9-base-pair direct target duplication. A novel kanamycin resistance transposon. Tn2681, can be generated from Tn2680 by IS903.B-mediated cointegration and subsequent reciprocal recombination between the directly repeated IS26 sequences. Tn2681 carries a single IS26 element in the middle of the transposon and is flanked by two directly repeated copies of IS903.B. Possible evolutionary relationships between Tn2680 and other kanamycin resistance transposons such as Tn903 and Tn2350 are discussed, based on the gene organization and DNA sequences. PMID- 2989254 TI - Characterization of Saccharomyces cerevisiae mutants supersensitive to aminoglycoside antibiotics. AB - We describe mutants of Saccharomyces cerevisiae that are more sensitive than the wild type to the aminoglycoside antibiotics G418, hygromycin B, destomycin A, and gentamicin X2. In addition, the mutants are sensitive to apramycin, kanamycin B, lividomycin A, neamine, neomycin, paromomycin, and tobramycin--antibiotics which do not inhibit wild-type strains. Mapping studies suggest that supersensitivity is caused by mutations in at least three genes, denoted AGS1, AGS2, and AGS3 (for aminoglycoside antibiotic sensitivity). Mutations in all three genes are required for highest antibiotic sensitivity; ags1 ags2 double mutants have intermediate antibiotic sensitivity. AGS1 was mapped 8 centimorgans distal from LEU2 on chromosome III. Analyses of yeast strains transformed with vectors carrying antibiotic resistance genes revealed that G418, gentamicin X2, kanamycin B, lividomycin A, neamine, and paromomycin are inactivated by the Tn903 phosphotransferase and that destomycin A is inactivated by the hygromycin B phosphotransferase. ags strains are improved host strains for vectors carrying the phosphotransferase genes because a wide spectrum of aminoglycoside antibiotics can be used to select for plasmid maintenance. PMID- 2989255 TI - Uncoupling of osmoregulation of the Escherichia coli K-12 ompF gene from ompB dependent transcription. AB - The expression of the genes for the Escherichia coli K-12 outer membrane proteins, ompF and ompC, is subject to osmoregulation and responds to changes in the osmolarity of the growth medium. The transcription of these genes is dependent on the products of the regulatory locus ompB (comprising the genes ompR and envZ). The native promoter of ompF was replaced with an inducible lpp promoter to eliminate this transcriptional dependence of ompF expression on ompB. As a result, it was possible for the OmpF protein to be produced in an ompB mutant strain that does not normally express ompF. Surprisingly, the expression of ompF under the lpp promoter was still osmoregulated not only in the ompB+ strain but also in two ompB strains tested. These results indicate the involvement of a factor(s) besides the ompR and envZ gene products in the osmoregulation of OmpF production. This factor may interact with a sequence downstream of the ompF promoter. In addition, we show that the expression of ompF under the lpp promoter has no direct effect on ompC expression. PMID- 2989256 TI - Cloning of phosphoenolpyruvate carboxylase gene from a cyanobacterium, Anacystis nidulans, in Escherichia coli. AB - The phosphoenolpyruvate carboxylase gene (ppc) from Anacystis nidulans, a cyanobacterium (blue-green alga), was cloned in Escherichia coli. Chromosomal DNA of A. nidulans was partially digested with Sau3AI, and the obtained DNA fragments were ligated in the BamHI site of pBR322. The hybrid plasmids were first transformed into E. coli K802 (hsdR-, hsdM+) to obtain the gene bank of A. nidulans. The bank consisted of about 12,000 clones. These hybrid plasmids were then transformed into E. coli PCR1 (ppc2-, recA1-, hsdR+, hsdM+), and the transformants were selected by complementation of the ppc mutation (phenotype of glutamate requirement). In the cell-free extracts of E. coli strains having the cloned ppc gene, PEPCase activities were detected, but their properties were different from those of the E. coli enzyme. Analysis by subcloning showed that the ppc gene was included in a DNA fragment 3,500 base pairs long and the maxicell method revealed that the molecular weight of the gene product was about 108,000. It is suggested that the ppc gene is expressed in E. coli mainly by read through transcription, being initiated by the promoter of tetracycline-resistance gene of pBR322, but the significant expression in reversed orientation of the cloned ppc gene indicates that the gene includes a promoter capable of functioning in E. coli cells. PMID- 2989257 TI - Efflux of preloaded iodide from the thyroid induced by externally added iodide. A study using a biological model of the thyroid iodide transport system. AB - Efflux of preloaded I- from the thyroid induced by externally added I- was studied using a biological model of the thyroid I- transport system. Phospholipid vesicles (P-vesicles) made from thyroid plasma membranes and soybean phospholipids were capable of accumulating I- in the presence of external Na+. P vesicles incubated in 136 mM Na+ containing 0.9 microM I- with 125I- for 2 min accumulated I- so that the I- concentration in the vesicles became about 2 microM. Addition of 5-20 microM stable I- to the incubation mixture at 2 min incubation resulted in a dose-dependent decrease in previously loaded 125I- in the vesicles. In other words, a dose-dependent increase in efflux of preloaded 125I- was observed. While the efflux occurred, Na+-dependent I- influx into P vesicles was preserved. When 2 mM ClO4-, a specific inhibitor of Na+-dependent I- influx, was added together with 10 microM I-, the external I- failed to diminish preloaded 125I- in P-vesicles. The 125I- efflux did not occur when a large amount of stable I- entered P-vesicles independently of Na+ in the presence of ClO4-. Similar 125I- efflux induced by externally added 5 microM SCN- was also blocked by simultaneously added ClO4-. These observations suggest that such I- efflux from the thyroid is a certain type of uphill I- transport which is closely related to Na+-dependent I- transport and that ClO4- and SCN- act on a common site of the I- transport system. PMID- 2989258 TI - Reciprocal changes of insulin and glucagon receptors in primary cultured hepatocytes. AB - The specific [125I]insulin binding to primary cultured hepatocytes was significantly greater than that to freshly isolated hepatocytes. Low affinity insulin binding sites in cultured cells were 6-fold greater in number than those of freshly isolated cells without a significant change in high affinity sites. However, both sensitivity (insulin concentration for half maximum stimulation) and responsiveness (% of increase above the basal level) to insulin for the stimulation of ODC activity were similar for isolated and cultured cells indicating an important role of high affinity sites in the insulin action. On the other hand, the specific [125I]glucagon binding to cultured cells was significantly decreased. Low affinity glucagon binding sites in cultured cells decreased by about 50% in cultured cells without a significant change in high affinity sites. Both sensitivity and responsiveness to glucagon for the stimulation of ketogenesis from palmitate also decreased as compared with those of isolated cells, indicating an important role of low affinity sites in the glucagon action. These results indicate that insulin and glucagon receptors were reciprocally changed in cultured cells, as compared with isolated cells. PMID- 2989261 TI - Do sodium and potassium forms of Na,K-ATPase differ in their secondary structure? AB - Infrared spectroscopy in the amide I region of purified membrane-bound Na,K ATPase preparation shows that Na+- and K+-bound forms of the enzyme have almost the same secondary structure. No difference is detected in the beta-structure (pleated sheets) content. This is contrary to the statement of the recent paper (Gresalfi, T. J., and Wallace, B. A. (1984) J. Biol. Chem. 259, 2622-2628) where a similar preparation was examined by circular dichroism spectroscopy and it was claimed that net 7% of protein peptide groups undergo a beta-sheet to alpha-helix conformational change upon Na,K-ATPase conversion from the K+ to the Na+ form. The discrepancy of the results is most likely caused by the particulate nature of the enzyme preparations used that could lead to optical artifacts in CD but not in IR measurements. A thorough comparison of IR spectra of these enzyme forms has revealed a very minor spectral difference which could suggest conformational perturbations, if any, of a much lower scale and another type than that claimed by Gresalfi and Wallace. The K+ form tends to absorb slightly more in the region of the alpha-helix band. This could reflect some distortion or a transition to a random coil structure of a small fraction of alpha-helical segments (less than or equal to 2% protein peptide groups) upon the enzyme conversion from the K+ to the Na+ form. PMID- 2989259 TI - Antimycin-resistant alternate electron pathway to plastocyanin in bovine-heart complex III. AB - Bovine-heart Complex III can catalyze the reduction of spinach plastocyanin by a decyl analog of ubiquinol-2 at a rate comparable with the rate of plastocyanin reduction by plastoquinol as catalyzed by the cytochrome b6-f complex purified from spinach leaves. This plastocyanin reduction as catalyzed by Complex III was almost completely inhibited by myxothiazol at stoichiometric concentrations, partially inhibited by UHDBT (5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole) and funiculosin, and was relatively insensitive to antimycin and HQNO (2-n-heptyl-4 hydroxyquinoline-N-oxide). Cytochrome c reduction as catalyzed by Complex III displayed a residual, inhibitor-insensitive rate of 5% of the uninhibited rate for each of the three inhibitors, antimycin, myxothiazol, and UHDBT. However, the residual rate that was insensitive to each of the inhibitors added singly was inhibited further by addition of the remaining two inhibitors. From these results it is concluded that plastocyanin reduction involves an electron-transfer pathway through Complex III that is distinct from the pathway utilized for reduction of cytochrome c. PMID- 2989260 TI - Divalent cations and the phosphatase activity of the (Na + K)-dependent ATPase. AB - Phosphatase activity of a kidney (Na + K)-ATPase preparation was optimally active with Mg2+ plus K+. Mn2+ was less effective and Ca2+ could not substitute for Mg2+. However, adding Ca2+ with Mg2+ or substituting Mn2+ for Mg2+ activated it appreciably in the absence of added K+, and all three divalent cations decreased apparent affinity for K+. Inhibition by Na+ decreased with higher Mg2+ concentrations, when Ca2+ was added, and when Mn2+ was substituted for Mg2+. Dimethyl sulfoxide, which favors E2 conformations of the enzyme, increased apparent affinity for K+, whereas oligomycin, which favors E1 conformations, decreased it. These observations are interpretable in terms of activation through two cases of cation sites. (i) At divalent cation sites, Mg2+ and Mn2+, favoring (under these conditions) E2 conformations, are effective, whereas Ca2+, favoring E1, is not, and monovalent cations complete. (ii) At monovalent cation sites divalent cations compete with K+, while Na+ at these sites favors E1 conformations. K+ increases the Km for substrate, but both Ca2+ and Mn2+ decrease it, perhaps by competing with K+. On the other hand, phosphatase activity in the presence of Na+ plus K+ is stimulated by dimethyl sulfoxide, by higher concentrations of Mg2+ and Mn2+, but not by adding Ca2+; this is consistent with stimulation occurring through facilitation of an E1 to E2 transition, perhaps an E1-P to E2-P step like that in the (Na + K)-ATPase reaction sequence. However, oligomycin stimulates phosphatase activity with Mg2+ plus Na+ alone or Mg2+ plus low K+: this effect of oligomycin may reflect acceleration, in the absence of adequate K+, of an alternative E2-P to E1 pathway bypassing the monovalent cation activated steps in the hydrolytic sequence. PMID- 2989262 TI - Regulation of glucagon receptor binding. Lack of effect of Mg and preferential role for GDP. AB - The effects of Mg2+ and guanine nucleotides on glucagon binding to its receptor were studied using [125I-Tyr10]monoiodoglucagon. Contrary to findings with beta adrenergic receptors, high affinity binding of the stimulatory hormone was not dependent on Mg2+ and low affinity binding could be obtained on nucleotide addition regardless of presence of Mg2+. GDP, guanyl-5'-yl thiophosphate (GDP beta S), GTP, and guanyl-5'-yl imidodiphosphate (GMP-P(NH)P) were all able to induce low affinity hormone binding. Since the Ns component of adenylyl cyclase, with which the receptor interacts, is inactive in stimulating the catalytic component C of adenylyl cyclase in the absence of Mg2+, both before and after GDP addition, it is suggested that Ns has at least two domains that change conformation independently of each other: a r domain, that interacts with the receptor and confers to it high affinity binding, and a c domain, that interacts with the catalyst C and stimulates it. It is suggested further that Ns is r+c- when stabilizing the receptor in its conformation with high affinity for hormone, and r-c- when under the influence of GDP which results in the receptor adopting the conformation that exhibits low affinity for the hormone. Comparison of potencies of the four nucleotides to induce low affinity binding showed that GDP and GDP beta S were equipotent and 10 times more potent than GTP and 100 times more potent than GMP-P(NH)P. Under the conditions used it was impossible to substantiate that the effects of GTP or GMP-P(NH)P were not due to formation of GDP from GTP or presence of GDP-like material in GMP-P(NH)P. It is suggested that, contrary to widely held opinions, GDP and GDP-like compounds, and not GTP or its analogs, are responsible for the lowering of the affinity of adenylyl cyclase stimulating receptors for their hormones or agonists. Furthermore, the experiments suggest that the c+ conformation of the c domain of Ns co-exists with the r+ and not the r- conformation of its r domain. PMID- 2989263 TI - A human platelet angiotensin I-processing system. Identification of components and inhibition of angiotensin-converting enzyme by product. AB - Mechanisms controlling the local generation of angiotensin II by vascular tissue are incompletely understood. Human platelets were examined for their ability to metabolize angiotensin I. Platelet-dependent angiotensin I metabolism was detected by a high performance liquid chromatography assay which allowed quantitation of angiotensin I substrate utilized and products formed. The major product of platelet-dependent angiotensin I metabolism was identified as des Leu10-angiotensin I. The platelet des-Leu10-angiotensin I-generating activity had a pH optimum of 6.0-6.5 and was inhibited 100% by mersalyl acid (10(-4) M), 86% by leupeptin (10(-4) M), and 95% by iodoacetamide (10(-2) M). The activity had an approximate Mr = 70,000 as determined by Sephacryl S-200 gel filtration. Intact human platelets stimulated with calcium ionophore (1-10 microM) released 13.7 30.8% of the des-Leu10-angiotensin I-generating activity. Des-Leu10-angiotensin I, the major product of platelet angiotensin I metabolism, inhibited human serum and purified rabbit lung angiotensin-converting enzymes with an I50 of 3.7 X 10( 6) and 2.0 X 10(-6) M, respectively. These results suggest that the platelet may control local angiotensin II formation at vascular sites both by metabolism of the precursor peptide angiotensin I and by generation of an endogenous angiotensin-converting enzyme inhibitor, des-Leu10-angiotensin I. This platelet dependent pathway may contribute to the control of local levels of vasoactive peptides, such as bradykinin and angiotensin II, so as to alter local tissue blood flow. PMID- 2989264 TI - Isolation of a phosphomonoesterase from human platelets that specifically hydrolyzes the 5-phosphate of inositol 1,4,5-trisphosphate. AB - Platelets, and a variety of other cells, rapidly hydrolyze the phosphoinositides in response to stimulation by agonists. One of the products of hydrolysis of phosphatidylinositol 4,5-diphosphate is inositol 1,4,5-trisphosphate, which recently has been suggested to mediate intracellular Ca2+ mobilization. We have found that human platelets contain an enzyme that degrades inositol 1,4,5 trisphosphate. We have isolated this soluble enzyme and find that it hydrolyzes the 5-phosphate of inositol 1,4,5-trisphosphate (Km = 30 microM, Vmax = 5.3 microM/min/mg of protein). The products of the reaction are inositol 1,4 diphosphate and phosphate. The apparent molecular weight of the enzyme is 38,000 as determined both by gel filtration and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and absence of 2-mercaptoethanol. This enzyme is specific for inositol 1,4,5-trisphosphate. Other water soluble inositol phosphates as well as phosphorylated sugars are not hydrolyzed, while the only inositol containing phospholipid hydrolyzed is phosphatidylinositol 4,5 diphosphate at a rate less than 1% that for inositol 4,5-trisphosphate. The inositol 1,4,5-trisphosphate 5-phosphomonoesterase requires Mg2+ for activity and is inhibited by Ca2+, Ki = 70 microM. Li+, up to 40 mM, has no effect on enzyme activity. The duration and magnitude of any inositol 1,4,5-trisphosphate response in stimulated platelets may be determined by the activity of this enzyme. PMID- 2989265 TI - Variability in transport rates of secretory glycoproteins through the endoplasmic reticulum and Golgi in human hepatoma cells. AB - We have previously shown that newly synthesized liver secretory proteins are exported at three distinct characteristic rates, with intracellular retention half-times of 110-120 min (e.g. transferrin), 75-80 min (e.g. ceruloplasmin), and 30-40 min (e.g. alpha 1-protease inhibitor) (J. B. Parent, H. Bauer, and K. Olden (1985) Biochim. Biophys. Acta, in press). In the present study we have determined the average time required for specific glycoproteins to move through the various compartments of the intracellular transport pathway, consisting of endoplasmic reticulum and Golgi complex. Localization in particular compartments was monitored by the use of the following complementary approaches: (i) Percoll density gradient fractionation of the subcellular organelles, (ii) sensitivity of the glycan moiety of N-linked glycosylation to endo-beta-N-acetylglucosaminidase H, and (iii) by the lectin-binding characteristics. The cell fractionation studies revealed that alpha 1-protease inhibitor, ceruloplasmin, and transferrin were transported from the rough endoplasmic reticulum with a retention half-time of 10, 30, or 45 min, respectively. Measurements of the rate at which newly synthesized glycoprotein became endo H-resistant (an event localized near the medial region of Golgi) demonstrated that it took 60-70, 30, and 18 min for 50% of transferrin, ceruloplasmin, and alpha 1-protease inhibitor, respectively, to reach the medial Golgi. Consistent with this finding, maximal binding of transferrin to wheat germ agglutinin (also a medial Golgi event) and Ricinus communis agglutinin I (a trans Golgi event) required 75 and 90 min, respectively, and maximal binding of ceruloplasmin to both lectins occurred in approximately 30 min. Maximal binding of alpha 1-protease inhibitor to wheat germ agglutinin and Ricinus communis agglutinin I required 15 and 30 min, respectively. The results presented here clearly indicate that (i) the time required for protein secretion cannot be entirely accounted for by lag in transport from the rough endoplasmic reticulum to the Golgi since the glycoproteins examined are retained in the former organelle for no more than two-fifths of the total intracellular retention half-time, and (ii) the variability in rates of protein secretion is not due solely to differences in rates of transport from the rough endoplasmic reticulum to the Golgi as variability in retention within the Golgi is also demonstrated. The results are discussed in terms of their compatibility with receptor-mediated transport of glycoproteins in both the endoplasmic reticulum and Golgi. PMID- 2989266 TI - Ubiquitin carboxyl-terminal hydrolase acts on ubiquitin carboxyl-terminal amides. AB - Ubiquitin carboxyl-terminal hydrolase (formerly known as ubiquitin carboxyl terminal esterase), from rabbit reticulocytes, has been shown to hydrolyze thiol esters formed between the ubiquitin carboxyl terminus and small thiols (e.g. glutathione), as well as free ubiquitin adenylate (Rose, I. A., and Warms, J. V. B. (1983) Biochemistry 22, 4234-4237). We now show that this enzyme hydrolyzes amide derivatives of the ubiquitin carboxyl terminus, including those of lysine (epsilon-amino), glycine methyl ester, and spermidine. It also hydrolyzes ubiquitin COOH-terminal hydroxamic acid, but is inactivated under the conditions for assaying ubiquitin-hydroxylamine adduct hydrolysis. Amide adducts formed between ubiquitin and epsilon-amino groups of protein lysine residues are much poorer substrates than is the ubiquitin amide of the epsilon-amino group of free lysine. The enzyme is thus a general hydrolase that recognizes the ubiquitin moiety, but is highly selective for small ubiquitin derivatives. It probably functions to regenerate ubiquitin from adventitiously formed ubiquitin amides and thiol esters. It also has the correct specificity to function in regenerating ubiquitin from small ubiquitin peptides that are probable end products of ubiquitin-dependent proteolysis. A simple, large-scale preparation of the enzyme from human erythrocytes is described. PMID- 2989267 TI - Fusion between Sendai virus envelopes and biological membranes. The use of fluorescent probes for quantitative estimation of virus-membrane fusion. AB - The fluorescent probes, N-4-nitrobenzo-2-oxa-1,3-diazole-phosphatidylethanolamine and lissamine-rhodamine-B-sulfonylphosphatidylethanolamine, were inserted at the appropriate surface density into membranes of reconstituted Sendai virus envelopes, thus allowing transfer of energy between the fluorescent probes. In addition, only the fluorescent molecule N-4-nitrobenzo-2-oxa-1,3-diazole phosphatidylethanolamine was inserted into the viral envelopes, resulting in self quenching. Incubation of fluorescent, reconstituted Sendai virus envelopes with human erythrocyte ghosts resulted in either reduction in the efficiency of energy transfer or in fluorescence dequenching. No reduction in the efficiency of energy transfer or fluorescence dequenching was observed when fluorescent, reconstituted Sendai virus envelopes were incubated with glutaraldehyde-fixed or desialized human erythrocyte ghosts. Similarly, no change in the fluorescence value was observed when nonfusogenic, reconstituted Sendai virus envelopes were incubated with human erythrocyte ghosts. These results clearly show that reduction in the efficiency of energy transfer or dequenching is due to virus-membrane fusion and not to lipid-lipid exchange. Incubation of reconstituted Sendai virus envelopes, carrying inserted N-4-nitrobenzo-2-oxa-1,3-diazolephosphatidylethanolamine, with cultured cells also resulted in a significant and measurable dequenching. However, incubation of nonfusogenic, fluorescent reconstituted Sendai virus envelopes with hepatoma tissue culture cells also resulted in fluorescent dequenching, the degree of which was about 50% of that observed with fusogenic, fluorescent reconstituted viral envelopes. It is therefore possible that, in addition to virus-membrane fusion, endocytosis of fluorescent viral envelopes results in fluorescence dequenching as well. PMID- 2989268 TI - Regulation of phosphatidylcholine biosynthesis in Chinese hamster ovary cells by reversible membrane association of CTP: phosphocholine cytidylyltransferase. AB - Treatment of Chinese hamster ovary cells with phospholipase C was previously shown to stimulate the CDP-choline pathway for phosphatidylcholine biosynthesis, and to cause activation of the CTP:phosphocholine cytidylyltransferase with a concomitant change in subcellular location of the enzyme (Sleight, R., and Kent, C. (1983) J. Biol. Chem. 258, 831-835). This paper presents a detailed analysis of the early events in the phospholipase C treatment, and provides evidence that the increased cytidylyltransferase activity causes the increased flux through the pathway. The time courses for the increase in cytidylyltransferase activity, increase in amount of membrane-associated enzyme, decrease in phosphocholine levels, and increase in phosphatidylcholine synthesis were similar, with all changes occurring within 30 min after addition of phospholipase C. These events preceded a decrease in cellular choline levels which correlated with a decreased capacity for choline uptake. The rate at which radioactive label was lost from pulse-labeled phosphocholine was the same as the rate at which label was incorporated into phosphatidylcholine, and these rates were stimulated 2.2-fold by phospholipase C treatment. We have also shown that the association of cytidylyltransferase with membranes was rapidly reversible when phospholipase C was removed from the cultures, and that the rate of decrease in phosphatidylcholine synthesis paralleled the rate of decrease in cytidylyltransferase activity. Cytidylyltransferase became reassociated with membranes when phospholipase C was added back to cultures from which it was previously removed. These results represent the first detailed account of the time frame involved in regulating phosphatidylcholine synthesis by the reversible association of cytidylyltransferase with cellular membranes. PMID- 2989269 TI - Fe(III).ATP complexes. Models for ferritin and other polynuclear iron complexes with phosphate. AB - Polynuclear iron complexes of Fe(III) and phosphate occur in seawater and soils and in cells where the iron core of ferritin, the iron storage protein, contains up to 4500 Fe atoms in a complex with an average composition of (FeO.OH)8FeO.OPO3H2. Although phosphate influences the size of the ferritin core and thus the availability of stored iron, little is known about the nature of the Fe(III)-phosphate interaction. In the present study, Fe-phosphate interactions were analyzed in stable complexes of Fe(III).ATP which, in the polynuclear iron form, had phosphate at interior sites. Such Fe(III).ATP complexes are important not only as models but also because they may play a role in intracellular iron transport and in iron toxicity; the complexes were studied by extended x-ray absorption fine structure, EPR, NMR spectroscopy, and measurement of proton release. Mononuclear iron complexes exhibiting a g' = 4.3 EPR signal were formed at Fe:ATP ratios less than or equal to 1:3, and polynuclear iron complexes (Fe greater than or equal to 250, EPR silent at g' = 4.3) were formed at an Fe:ATP ratio of 4:1. No NMR signals due to ATP were observed when Fe was in excess (Fe:ATP = 4:1). Extended x-ray absorption fine structure analysis of the polynuclear Fe(III).ATP complex was able to distinguish an Fe-P distance at 3.27 A in addition to the octahedral O at 1.95 A and 4-5 Fe atoms at 3.36 A. The Fe-O and Fe-Fe distances are the same as in ferritin, and the Fe-P distance is analogous to that in another metal-ATP complex. An observable Fe-P environment in such a large polynuclear iron cluster as the Fe(III).ATP (4:1) complex indicates that the phosphate is distributed throughout rather than merely on the surface, in contrast to earlier models of chelate-stabilized iron clusters. Complexes of Fe(III) and ATP similar to those described here may form in vivo either as normal components of intracellular iron metabolism or during iron excess where the consequent alteration of free nucleotide triphosphate pools could contribute to the observed toxicity of iron. PMID- 2989270 TI - Gene structure of a major form of phenobarbital-inducible cytochrome P-450 in rat liver. AB - The gene structure of cytochrome P-450b, a major form of phenobarbital-inducible cytochrome P-450 in rat livers was elucidated by sequence analysis of the cloned genomic DNAs and was compared with the previously determined gene structures of cytochrome P-450e, a minor form of phenobarbital-inducible cytochrome P-450 and two forms of 3-methylcholanthrene-inducible cytochrome P-450 (P-450c and -d). The gene for cytochrome P-450b is 23 kilobase pairs (kb) long and is separated into 9 exons by 8 intervening sequences. This gene structure is very similar to that of cytochrome P-450e except for the first intron, the first intron being much longer in cytochrome P-450b gene (approximately 12 kb) than in cytochrome P-450e gene (3.2 kb), but differs greatly from the gene structures of two 3 methylcholanthrene-inducible cytochrome P-450s as pointed out previously (Sogawa, K., Gotoh, O., Kawajiri, K. & Fujii-Kuriyama, Y. (1984) Proc. Natl. Acad. Sci. U.S.A. 81, 5066-5070). The nucleotide sequences in all 9 exons and their flanking regions in introns show very close homology between the two phenobarbital inducible cytochrome P-450 genes. Forty base substitutions are found in approximately 1900 nucleotides of all exonic sequences, and 15 of them result in 14 amino acid replacements. These base substitutions occur in relatively limited regions of the gene sequences. Most of them are found in exons 6, 7, 8, and 9, most frequently in exon 7 as described previously (Mizukami, Y., Sogawa, K., Suwa, Y., Muramatsu, M. & Fujii-Kuriyama, Y. (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 3958-3962). The close sequence homology between the two phenobarbital inducible cytochrome P-450 genes is also found to extend to the promoter region with one notable exception. The simple repeated sequences of (CA)n which is present at -254 position in cytochrome P-450e gene is also observed at the equivalent position in cytochrome P-450b gene, but the repetitiveness is greatly reduced in cytochrome P-450b gene ((CA)5 for P-450b versus (CA)19 for P-450e), and this may somehow be related to the difference in the level of cytochrome P 450b and P-450e in the inductive phase of phenobarbital administration. PMID- 2989271 TI - Regulation of protein kinase and its regulatory subunits during skeletal myogenesis. AB - Rat skeletal myoblasts contain two cytosolic cAMP-dependent protein kinases, types I and II. Photoaffinity labeling with 8-azido-cAMP reveals the presence of regulatory subunits of Mr = 52,000, 47,000, and 36,000. The Mr = 52,000 and 47,000 subunits are very likely RII and RI, respectively, while the Mr = 36,000 subunit appears to be a proteolytic product of RI, as judged by its cross reactivity to anti-RI antiserum. The total protein kinase activity increases about 3-fold during the fusion of myoblasts. In parallel with this increase, the concentration of RI subunit also increases, while the levels of RII remain unchanged. Myoblast mutants which lack the capability to differentiate both biochemically and morphologically also lack the ability to increase the concentration of RI subunit. This ability is restored in complementing somatic hybrids which regain the capability to differentiate. PMID- 2989272 TI - The Na+/H+ antiport is activated by serum and phorbol esters in proliferating myoblasts but not in differentiated myotubes. Properties of the activation process. AB - The properties of the Na+/H+ exchange system have been studied with 22Na+ uptake techniques at two stages of muscle development: proliferating myoblasts and differentiated myotubes. The characteristics of the interactions of the exchanger with external H+, with external Na+, and with amiloride or its more potent analogs are the same at both stages of development. Differences between the two stages of development concern: (i) the internal pH (pHi) dependence of the Na+/H+ exchanger, and (ii) the activation of the Na+/H+ exchanger by serum and phorbol ester which is observed in myoblasts but not in myotubes. Properties of the Na+/H+ exchanger in myoblasts after serum activation seem to be identical to those observed in myotubes with or without serum as if myotube formation stabilized a fully activated state of the exchanger. The activation of the myoblast Na+/H+ exchange system by serum is due to a shift of the pHi dependence towards alkaline pHi values and to an increase in the maximal activity of the Na+/H+ exchange system at acidic pH. Phorbol esters which are well-known activators of protein kinase C can only partially mimic the effects of serum on the Na+/H+ exchanger: they produce a shift of the pH dependence, but they do not increase the maximal activity at acidic pH. PMID- 2989274 TI - Characterization of the human melanoma nerve growth factor receptor. AB - Monoclonal antibodies to the human nerve growth factor receptor have been used to biochemically characterize the receptor in the human melanoma cell line A875. Labeling of A875 cell proteins by culture with [35S]cysteine or labeling of cell surface proteins with 125I followed by immunoprecipitation with anti-nerve growth factor receptor antibody reveals a receptor protein with an apparent Mr of 70,000 75,000 and an isoelectric point of 4.9-5.2. Incorporation of [3H]glucosamine into this species indicates it is a glycoprotein. The receptor becomes phosphorylated on serine residues in intact cells and in isolated membranes incubated with [gamma-32P]ATP. The receptor appears to exist, at least partially, in the form of a disulfide-linked oligomer (probably a dimer) of Mr = 75,000 subunits. Kinetic [35S]cysteine labeling studies reveal an Mr = 59,000 core protein which is glycosylated via N-linked and probably also O-linked sugar moieties to produce the mature (Mr = 70,000-75,000) receptor. PMID- 2989273 TI - Phorbol ester causes desensitization of gonadotropin-responsive adenylate cyclase in a murine Leydig tumor cell line. AB - The murine Leydig tumor cell line, MLTC-1, contains gonadotropin receptors and a gonadotropin-responsive adenylate cyclase system that became refractory (desensitized) when exposed to human chorionic gonadotropin (hCG). MLTC-1 cells also contain phorbol ester receptors with a Kd of 53 nM for [3H]phorbol dibutyrate. Exposing cells to 12-O-tetradecanoyl phorbol 13-acetate (TPA) also causes desensitization of the hCG response. TPA-induced desensitization was similar to hCG-induced desensitization by every criteria tested. Both TPA- and hCG-induced desensitization caused approximately 50% loss of the hormone response within 30 min. Neither TPA or hCG altered receptor affinity for hCG. The dose response of adenylate cyclase to hCG or GTP in isolated membranes was not affected by either hCG- or TPA-induced desensitization. Similarly the dose response to hCG of cAMP accumulation in intact cells was not altered by desensitization with hCG or TPA. It was determined that MLTC-1 cells have Ca2+/phospholipid-dependent protein kinase activity that displayed a dose dependent response to TPA. The concentration of TPA required to activate the protein kinase was similar to that required for desensitization. Phorbol esters that were unable to activate protein kinase C were also unable to desensitize MLTC-1 cells. The protein kinase from MLTC-1 cells was also activated by diacylglycerol. In addition, diacylglycerols caused desensitization of the hCG response. TPA- and diacylglycerol-induced desensitization is probably mediated by protein kinase C, and the similarities between hCG- and TPA-induced refractoriness suggests a convergence of mechanisms at some point of MLTC-1 cell desensitization. PMID- 2989275 TI - Purification and characterization of a protein-tyrosine kinase encoded by the Abelson murine leukemia virus. AB - Sequences termed v-abl, which encode the protein-tyrosine kinase activity of Abelson murine leukemia virus, have been expressed in Escherichia coli as a fusion product (ptabl50 kinase). This fusion protein contains 80 amino acids of SV40 small t and the 403 amino acid protein kinase domain of v-abl. We report here the purification and characterization of this kinase. The purified material contains two proteins (Mr = 59,800 and 57,200), both of which possess sequences derived from v-abl. Overall purification was 3,750-fold, with a 31% yield, such that 117 micrograms of kinase could be obtained from 40 g of E. coli within 6-7 days. The specific kinase activity is over 170 mumol of phosphate min-1 mumol-1, comparable to the most active protein-serine kinases. Kinase activity is insensitive to K+, Na+, Ca2+, Ca2+-calmodulin, cAMP, or cAMP-dependent protein kinase inhibitor. The Km for ATP is dependent on the concentration of the second substrate. GTP can also be used as a phosphate donor. The enzyme can phosphorylate peptides consisting of as few as two amino acids and, at a very low rate, free tyrosine. Incubation of the kinase with [gamma-32P]ATP results in incorporation of 1.0 mol of phosphate/mol of protein. This reaction, however, cannot be blocked by prior incubation with unlabeled ATP. Incubation of 32P labeled kinase with either ADP or ATP results in the synthesis of [32P]ATP. This suggests the phosphotyrosine residue on the Abelson kinase contains a high energy phosphate bond. PMID- 2989276 TI - Two mechanisms for inhibition of ADP-induced platelet shape change by 5'-p fluorosulfonylbenzoyladenosine. Conversion to adenosine, and covalent modification at an ADP binding site distinct from that which inhibits adenylate cyclase. AB - The interaction of ADP with platelets leads to shape change, exposure of fibrinogen binding sites, and aggregation, all of which have been shown to be inhibited by 5'-p-fluorosulfonylbenzoyladenosine (FSBA), an alkylating analogue of adenine nucleotides which binds covalently to a 100-kDa polypeptide in intact platelet membranes (Figures, W. R., Niewiarowski, S., Morinelli, T., Colman, R. F., and Colman, R. W. (1981) J. Biol. Chem. 256, 7789-7795). In plasma, FSBA can break down to adenosine which stimulates adenylate cyclase. To distinguish between direct effects of FSBA and the actions of adenosine, we have used washed platelet suspensions and adenosine deaminase. We studied the effects of FSBA on shape change and cyclic AMP metabolism, and on the binding of 2-methylthio-ADP, which mimics the effects of ADP on cyclic AMP metabolism at concentrations too low to activate platelets. Inhibition of ADP-induced shape change of platelets incubated with FSBA for 2 min in platelet-rich plasma was greatly reduced by adenosine deaminase. In the presence of a phosphodiesterase inhibitor, 100 microM FSBA increased platelet cyclic AMP to the same extent as did 10 microM adenosine. These effects were inhibited by theophylline, an adenosine receptor antagonist, and by adenosine deaminase. Incubation of washed platelets for 60 min with FSBA and adenosine deaminase caused a concentration-dependent inhibition of ADP induced shape change. Inhibition closely paralleled the covalent incorporation of 3H from tritiated FSBA into platelet membranes. Under these conditions, FSBA did not block inhibition of cyclic AMP accumulation by ADP, nor did it block the binding of 2-methylthio-ADP. We conclude that part of the inhibition of shape change caused by brief exposure to FSBA is due to adenosine, but at longer times shape change is inhibited in association with covalent incorporation of sulfonylbenzoyladenosine. This effect of FSBA is independent of adenosine and occurs at a site distinct from that at which ADP inhibits adenylate cyclase. PMID- 2989277 TI - Characterization and identification of the hyaluronate binding site from membranes of SV-3T3 cells. AB - Previous research has shown that binding sites for hyaluronate are present on the surfaces of a number of different cell types. To further characterize these binding sites, membranes were prepared from SV-3T3 cells and dissolved in a solution of sodium deoxycholate. Hyaluronate binding activity was detected by mixing the sodium deoxycholate extract with [3H]hyaluronate and then adding an equal volume of saturated (NH4)2SO4, which precipitated the binding protein and any [3H]hyaluronate associated with it, but left free [3H]hyaluronate in solution. Following partial purification by hydroxylapatite chromatography, the binding site was examined by molecular sieve chromatography and by rate-zonal centrifugation, which revealed that it has a Stokes radius of 6.5 nm and a sedimentation coefficient of 4.8 S. From these values, it was possible to calculate that the sodium deoxycholate-solubilized binding site has a frictional coefficient of 1.87 and a molecular weight of 132,000. Since this latter value applies to the complex of both detergent and protein, the binding protein by itself must have a molecular weight lower than 132,000. To determine the molecular weight of the hyaluronate binding site itself, the protein was purified by the sequential application of hydroxylapatite chromatography, molecular sieve chromatography, rate-zonal centrifugation, and finally lectin-affinity chromatography on concanavalin A-agarose. Analysis of the purified material by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed an 85,000 Mr protein which has been identified as the binding site. This protein was also detected on nitrocellulose blots which had been specifically stained for concanavalin A binding material, suggesting that the binding site is a glycoprotein. PMID- 2989278 TI - Distribution of a fatty acid spin probe in sarcoplasmic reticulum. Evidence of membrane asymmetry. AB - The distribution of a lipophilic spin probe, 5-doxyl stearate, between the inner and outer halves of the sarcoplasmic reticulum (SR) bilayer was determined by titration with Ni X EDTA, a spin broadening agent. Titrations were also performed with Fe(CN)3-6 and with the solvated Ni2+ cation. Ni X EDTA titrations reached a clearly defined asymptote at 35% signal reduction. Fe(CN)3-6 and Ni2+ titrations gave biphasic curves but showed 35% of the signal to be readily eliminated at low concentrations. When the Ni2+ cation was used with ionophore, titrations indicated that 96% of the probe is aligned in the bilayer with the spin moiety at either the inner or outer interface. It was concluded that the spin probe distribution between the outer and inner halves of the SR bilayer is 35:65, respectively. Titrations performed on vesicles of purified SR lipids gave a ratio of 60 exposed:40 protected, consistent with the vesicular geometry. In addition the spin probe distribution in SR vesicles did not vary as a function of temperature, salt concentration, or spin probe concentration. On this basis it was concluded that the spin probe distribution gives a reasonable estimation of the volume of fluid lipids available to readily solubilize the probe in each half of the bilayer and that the observed asymmetry in distribution is due to the presence of SR proteins which were eliminated in the pure lipid vesicles. Furthermore, as EDTA is unique in its ability to chelate transition metals, Ca2+ and EGTA can be used in Ni X EDTA titrations without altering the chelation of Ni2+. Known changes in ATPase conformation accompanying Ca2+ and adenyl-5'-yl imidodiphosphate X Mg binding did not affect the spin probe distribution. However, phosphorylation of the enzyme by Pi gave a small, but clearly discernible, protection of spin probe signal. Chemical reduction with ascorbate indicated that this was due to occlusion of a small fraction of spin probes and thus possibly SR lipids. PMID- 2989279 TI - A water-soluble form of porin from the mitochondrial outer membrane of Neurospora crassa. Properties and relationship to the biosynthetic precursor form. AB - Mitochondrial porin, the outer membrane pore-forming protein, was isolated in the presence of detergents and converted into a water-soluble form. This water soluble porin existed under nondenaturing conditions as a mixture of dimers and oligomers. The proportion of dimers increased with decreasing porin concentration during conversion. Water-soluble porin inserted spontaneously into artificial bilayers as did detergent-solubilized porin. Whereas the latter form had no specific requirements for the lipid composition of the bilayer, water-soluble porin inserted only into membranes containing a sterol, and only in the presence of very low concentrations of Triton X-100 (0.001% w/v) in the solution bathing the bilayer. The channels formed by water-soluble porin were indistinguishable from those formed by detergent-purified porin with respect to specific conductance and voltage dependence of conductance. Water-soluble porin bound tightly in a saturable fashion to isolated mitochondria. The bound form was readily accessible to added protease, indicating its presence on the mitochondrial surface. The number of binding sites was in the range of 5-10 pmol/mg of mitochondrial protein. Water-soluble porin apparently binds to a site on the assembly pathway of the porin precursor, since mitochondria whose binding sites were saturated with the water-soluble form did not import porin precursor synthesized in a cell-free system. PMID- 2989280 TI - An equilibrium between distorted and undistorted DNA in the adult chicken beta A globin gene. AB - We have used single strand specific nucleases to map DNA distortion in the adult chicken beta A-globin gene. We have detected two structures of that kind and have mapped nuclease-cutting sites at one base resolution. One prominent site is centered at -190 relative to the RNA capping site and is positioned at the center of a stretch of contiguous C residues. The second site is near the first intron/exon junction (+620) and appears as a series of discrete 1-base-long enzyme-cutting sites. Based upon the pattern of nuclease cutting and the kinetics of nuclease cutting we conclude that the "poly(C)" stretch may assume a looped geometry in supertwisted DNA molecules which is similar to that proposed by Felsenfeld (Nickol, J. M., and Felsenfeld, G. (1983) Cell 35, 467-477). We show that S1 nuclease cuts within the intron occur mainly at the end points of polypurine segments and suggest that such end points may assume a distorted transitional geometry. We find that Neurospora crassa endonuclease cuts both the promotor and intron sites in linear DNA molecules but that in linear DNA the cutting process is limited by a first order conformation change of the DNA substrate. Based upon those kinetics we propose that in unstressed DNA, each of the two sites can convert between a distorted and undistorted geometry. In the enzyme assay buffer at 37 degrees C, the time constant for the equilibrium is nearly 10 h for the promotor site and 7 h for the intron. PMID- 2989281 TI - Evolution and heterogeneity of the alpha-/beta-type and gamma-type gliadin DNA sequences. AB - Near full length cDNA clones for both alpha-/beta- and gamma-type gliadins were isolated and studied for sequence diversity. Based on restriction site polymorphism and cross-hybridization studies, alpha-/beta- and gamma-type clones could be divided into five and three homology classes, respectively. Clones representing each of the different classes were sequenced and compared. Sequence divergence between the classes was due to single-base substitutions and to duplications or deletions within or near direct repeats. Thus, through numerous duplications and subsequent divergence, the gliadin multigene family encodes a polymorphic set of polypeptides differing in both isoelectric point and molecular size. Southern blot analysis of wheat DNA suggested that the number of genes encoding the alpha-/beta-type gliadins was extremely large (greater than 100 copies/haploid genome). Inasmuch as hybridization patterns were the same using DNA isolated from seeds or leaves, amplification or rearrangement of DNA does not occur during development. The complete coding sequence of a gamma-gliadin was similar to that observed for the alpha-/beta-gliadins, but with several notable differences. Comparison of gamma-type gliadin cDNA sequences showed that, unlike the conserved dodecamer repeat common to all the alpha-/beta-gliadins, the tandem repeat unit differed among gamma-gliadin clones. PMID- 2989282 TI - Structure of two unlinked Drosophila melanogaster glyceraldehyde-3-phosphate dehydrogenase genes. AB - Two Drosophila genes that code for the enzyme glyceraldehyde-3-phosphate dehydrogenase (Gapdh) have been isolated and their structures determined by DNA sequence analysis. The two genes, Gadph-1 and Gapdh-2, are homologous to each other in their coding regions but differ entirely in the 5' and 3' flanking regions. Both genes are functionally expressed in adult flies as determined by Northern blot analysis using gene-specific probes. Gapdh-1 is mapped by in situ hybridization at position 43E-F on the right arm of the second chromosome and Gapdh-2 at position 13F on the left arm of the X chromosome. Transcription initiation sites as well as polyadenylation sites for both Gapdh transcripts have also been determined. Gapdh-1 lacks a sequence homologous to the TATA box in its 30-base pair region that is characteristic of many RNA polymerase II transcribed promoters. In contrast, Gapdh-2 contains a consensus TATA box sequence as well as a CAAT box in its promoter region. Furthermore, a sequence element ATTTGCAT (dc) and nontandem multiple direct repeats have been found in the -35 to -155-base pair 5' flanking region. Other than the intron located in the 5' noncoding region of Gapdh-2, both genes lack intervening sequences. PMID- 2989283 TI - Photoaffinity labeling of atrial natriuretic factor receptors of rat kidney cortex plasma membranes. AB - Synthetic rat atrial natriuretic factor (ANF) was derivatized with the N hydroxysuccinimide ester of [125I]iodoazidosalicylic acid to yield a radioactive photoaffinity probe. Incubation of purified plasma membranes from rat kidney cortex with this photoaffinity probe resulted in the specific labeling of a 140 kDa glycoprotein. The photoaffinity labeling of this protein was inhibited by ANF but not by reduced and alkylated ANF nor by other unrelated peptides. A 140-kDa band was also specifically labeled in liver plasma membranes but not in adipocyte plasma membranes. These observations suggest strongly that the 140-kDa glycoprotein is the ANF receptor. PMID- 2989285 TI - The effect of chloride on the redox and EPR properties of myeloperoxidase. AB - Myeloperoxidase was purified from human polymorphonuclear leukocytes and the effect of chloride upon the EPR and potentiometric properties was studied. The redox titration between the ferrous and ferric states of the enzyme yielded n = 1 Nernst plots between pH 9 and 4, with clear isosbestic points in the optical spectra during the redox change. The midpoint potential (Em) between the ferric and ferrous forms of the enzyme exhibited a pH-dependent change between pH 4 and 9, and the effect of added chloride ion indicated that Cl- competed with OH- for a binding site on the enzyme. Interestingly, the pH dependence of the Em indicated that the overall redox reactions of the enzyme was: ferric myeloperoxidase + 2e- + 1H+ = ferrous myeloperoxidase. Myeloperoxidase exhibited a rhombic high spin EPR signal which exhibited reduced rhombicity upon the binding of chloride. Our results strongly suggest that chloride binds to the sixth coordination position of the chlorin iron in myeloperoxidase by replacing the water which is the sixth ligand in the resting state. It is also concluded that the two iron centers are identical and that there is no interaction between them. PMID- 2989284 TI - Structural characterization of the avian retrovirus reverse transcriptase and endonuclease domains. AB - The enzymatic domains of the avian retrovirus polymerase (pol) gene have been mapped by the use of peptide antibodies and COOH-terminal amino acid analysis. The processed pol beta polypeptide is cleaved in vivo to yield alpha and pp32. Rabbit antibodies were directed against synthetic peptides whose sequence was deduced from the known pol sequence of Rous sarcoma virus, Prague C (Schwartz, D.E., Tizard, R., and Gilbert, W. (1983) Cell 32, 853-869). The RNase H active site of pol was located in the NH2-terminal region of the alpha DNA polymerase subunit. The COOH terminus of the alpha subunit was found to be immediately adjacent to the NH2 terminus of the pp32 pol protein. COOH-terminal amino acid analysis of pp32 revealed that this protein is also processed. From the deduced amino acid sequence of pol, it appears likely that pol encodes an additional 4100 dalton polypeptide located at its extreme COOH terminus. The enzymatic domains on beta appear to map in the following order: RNase H-DNA polymerase-DNA endonuclease. Hydrophilicity analysis and secondary structure predictions of wild type Rous sarcoma virus pol products and mutated pp32 possessing single amino acid changes permit further structural evaluation of the multifunctional pol protein. PMID- 2989286 TI - Purification and characterization of a guanosine diphosphatase activity from calf liver microsomal salt wash proteins. AB - A potent guanosine diphosphatase activity that hydrolyzes GDP to 5'-GMP + Pi has been isolated and purified from the salt wash proteins of calf liver microsomes. The purified enzyme, a monomeric protein of approximate Mr 46,000, possesses nucleotide substrate specificity since, among the nucleoside diphosphates and triphosphates tested, only GDP and UDP are hydrolyzed by the enzyme. The relative affinity of the enzyme for GDP is, however, much higher than for UDP. The effect of the enzyme on the binary complex formed between eukaryotic initiation factor 2 (eIF-2) and GDP has also been investigated. The enzyme neither hydrolyzes GDP bound to eIF-2 nor catalyzes the exchange of eIF-2-bound GDP with GTP even in the presence of Met-tRNAf. The enzyme, therefore, is presumably not involved in recycling of eIF-2 in eukaryotic polypeptide chain initiation reaction. The possible biological function of the enzyme in maintaining the cellular pool of GTP-GDP is discussed. PMID- 2989287 TI - Amino acid sequence of the active site peptide of bovine intestinal 5'-nucleotide phosphodiesterase and identification of the active site residue as threonine. AB - We report here the identification of the amino acid residue which forms the covalent intermediate in the catalytic mechanism of bovine intestinal 5' nucleotide phosphodiesterase and the sequence of the neighboring amino acids. The active site of 5'-nucleotide phosphodiesterase was labeled using thymidine 5' [alpha-32P]triphosphate as substrate. A single labeled cyanogen bromide peptide was isolated using reversed-phase high performance liquid chromatography. After subdigestion with endoproteinase Lys-C and chymotrypsin, the entire amino acid sequence of the 60-residue active site peptide was obtained using automated Edman degradation. All of the radioactivity of the active site peptide was localized to a hexapeptide with sequence Thr-Phe-Pro-Asn-His-Tyr. Phosphoamino acid analysis of this peptide indicated that the labeled residue was threonine. We are not aware of any other enzymes in which threonine is phosphorylated as a covalent intermediate in the catalytic mechanism. PMID- 2989288 TI - Mechanism of oxidative C alpha-C beta cleavage of a lignin model dimer by Phanerochaete chrysosporium ligninase. Stoichiometry and involvement of free radicals. AB - The hemoprotein ligninase of Phanerochaete chrysosporium Burds. catalyzes the oxidative cleavage of lignin model dimers between C alpha and C beta of their propyl side chains. The model dimers hitherto used give multiple products and complex stoichiometries upon enzymatic oxidation. Here we present experiments with a new model dimer, 1-(3,4-dimethoxyphenyl)-2-phenylethanediol (dimethoxyhydrobenzoin, DMHB) which is quantitatively cleaved by ligninase in air to give benzaldehyde and veratraldehyde according to the stoichiometry: 2DMHB + O2----2PhCHO + 2Ph(OMe)2CHO. Catalytic amounts of H2O2 are required for this aerobic reaction. Under anaerobic conditions, ligninase uses H2O2 as the oxidant for cleavage: DMHB + H2O2----PhCHO + Ph(OMe)2CHO. Electron spin resonance experiments done in the presence of spin traps, 2-methyl-2-nitrosopropane or 5,5 dimethyl-1-pyrroline-N-oxide, show that C alpha-C beta cleavage yields alpha hydroxybenzyl radicals as intermediate products. Under anaerobic conditions, these radicals react further to give the final aldehyde products. In air, O2 adds to the carbon-centered radicals, probably giving alpha-hydroxybenzylperoxyl radicals which fragment to yield superoxide, benzaldehyde, and veratraldehyde. These results lead us to propose a mechanism for C alpha-C beta cleavage in which attack by ligninase and H2O2 on the methoxylated ring of DMHB yields a cation radical, which then cleaves to give either benzaldehyde and an alpha hydroxy(dimethoxybenzyl) radical or veratraldehyde and an alpha-hydroxybenzyl radical (cf. Kersten, P. J., Tien, M., Kalyanaraman, B., and Kirk, T.K. (1985) J. Biol. Chem. 260, 2609-2612; Snook, M. E., and Hamilton, G. A. (1974) J. Am. Chem. Soc. 96, 860-869). Similar mechanisms probably apply to the enzymatic C alpha-C beta cleavage of natural lignin. PMID- 2989289 TI - Diffusion of extracellular hydrogen peroxide into intracellular compartments of human neutrophils. Studies utilizing the inactivation of myeloperoxidase by hydrogen peroxide and azide. AB - It is well known that catalase is transformed to nitric oxide-Fe2+-catalase by hydrogen peroxide (H2O2) plus azide. In this report, we show that myeloperoxidase is also inactivated by H2O2 plus azide. Utilizing this system, we studied the presence and source of intracellular H2O2 generated by activated neutrophils. Stimulation of neutrophils with phorbol myristate acetate (PMA, 100 ng/ml) plus azide (5 mM) for 30 min completely inactivated intragranular myeloperoxidase and reduced cytosolic catalase to 35% of resting cells. This intracellular inactivation of heme enzymes did not occur in normal neutrophils incubated with either PMA or azide alone or in neutrophils from patients with chronic granulomatous disease (CDG) which cannot produce H2O2 in response to PMA. Incubation of neutrophils with azide and a H2O2 generating system (glucose glucose oxidase) inactivated 41% of neutrophil myeloperoxidase. Glutathione glutathione peroxidase (GSH-GSH peroxidase), an extracellular H2O2 scavenger, totally protected neutrophil myeloperoxidase from inactivation by azide plus glucose-glucose oxidase. In addition, when a mixture of normal and CGD cells was stimulated with PMA in the presence of azide, 90% of the myeloperoxidase in CGD neutrophils was inactivated. Therefore, H2O2 released extracellularly from activated neutrophils can diffuse into cells. In contrast, myeloperoxidase in normal polymorphonuclear leukocytes stimulated with PMA in the presence of azide and GSH-GSH peroxidase was 75% inactivated. Thus, the results indicate that a GSH GSH peroxidase-insensitive pool of H2O2 is also generated, presumably at the plasma membrane, and this pool of H2O2 can undergo direct internal diffusion to inactivate myeloperoxidase. PMID- 2989290 TI - Nucleotide sequence of yeast GDH1 encoding nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase. AB - The yeast GDH1 gene encodes NADP-dependent glutamate dehydrogenase. This gene was isolated by complementation of an Escherichia coli glutamate auxotroph. NADP dependent glutamate dehydrogenase was overproduced 6-10-fold in Saccharomyces cerevisiae bearing GDH1 on a multicopy plasmid. The nucleotide sequence of the 1362-base pair coding region and 5' and 3' flanking sequences were determined. Transcription start sites were located by S1 nuclease mapping. Regulation of GDH1 was not maintained when the gene was present on a multicopy plasmid. Protein secondary structure predictions identified a region with potential to form the dinucleotide-binding domain. The amino acid sequences of the yeast and Neurospora crassa enzymes are 63% conserved. Unlike the N. crassa gene, yeast GDH1 has no introns. PMID- 2989291 TI - Characterization of receptors for immune interferon in U937 cells with 32P labeled human recombinant immune interferon. AB - Recombinant human immune interferon (HuIFN-gamma) was labeled with [gamma-32P]ATP and cyclic-AMP-dependent protein kinase from bovine heart to a specific radioactivity of 11,000 Ci/mmol. At least two molecules of phosphate were incorporated per molecule of interferon. The binding of [32P]HuIFN-gamma to human U937 histiocytic lymphoma cells was time dependent, and displaceable by HuIFN gamma but not by HuIFN-alpha A or HuIFN-beta. The specific binding was saturable with less than 10% nonspecific binding. The dissociation constant of [32P]HuIFN gamma for U937 interferon receptors was calculated to be 1.5 X 10(-10) M with a total of 1,800 binding sites/cell. Dissociation of bound [32P]IFN-gamma at 24 degrees C exhibited two distinct rates. A fast dissociation with a specific rate constant of 0.141 min-1, and a slow dissociation with a specific rate constant of 0.0027 min-1. The Kd for [32P]HuIFN-gamma was calculated from kinetic constants to be 5.4 X 10(-10) M. PMID- 2989292 TI - Purification of pT181-encoded repC protein required for the initiation of plasmid replication. AB - The plasmid pT181 of Staphylococcus aureus consists of 4437 base pairs and encodes resistance to tetracycline. Initiation of pT181 replication specifically requires the plasmid-encoded repC protein. An in vitro system has been shown to carry out semiconservative replication of pT181 and its derivative plasmids (Khan, S A., Carleton, S. M., and Novick, R. P. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 4902-4906). We have used this replication assay to isolate repC protein, which was purified to near homogeneity. The repC gene was cloned into the pKJB825 plasmid that contains the phage lambda temperature-sensitive repressor gene, cI857, and the rightward promoter, PR. Upon temperature induction, Escherichia coli clones containing the recombinant plasmid overproduced repC protein, which was purified in significant quantities. The molecular weight of repC protein under denaturing conditions is 38,000, which is consistent with the size predicted from the DNA sequence data. Presence of repC protein was absolutely essential for the initiation of replication of pT181 and its derivatives in vitro. PMID- 2989293 TI - Cloning and sequence analysis of cDNAs for human high molecular weight and low molecular weight prekininogens. Primary structures of two human prekininogens. AB - cDNA sequences for both human high molecular weight (HMW) and low molecular weight (LMW) prekininogens have been isolated by molecular cloning and determined by sequence analysis. The sequence determination together with the S1 nuclease mapping and RNA blot-hybridization analyses indicate that human HMW and LMW prekininogen mRNAs share an identical sequence throughout the 5'-untranslated region and the protein-coding region up to the sequence encoding the 12 amino acids distal to the bradykinin sequence, and the two mRNAs then completely diverge from each other. The signal peptide, the heavy chain (H chain), and the bradykinin moiety, which are common between the two prekininogens, consist of 18, 362, and 9 amino acids, respectively, while the light chains (L chains) of the HMW and LMW prekininogens are composed of 255 and 38 amino acids, respectively. All 17 cysteine residues present in the human and bovine H chains are located at exactly equivalent positions, indicating that the human H chain, like the bovine counterpart, can form 8 loop structures, each connected by two adjacent cysteine residues. The L chains of human and bovine kininogens differ in the protein lengths as well as in some amino acids crucial for the processing of the kininogens by kallikrein. Based upon this finding, we have discussed the molecular basis for the different modes of processing of human and bovine HMW kininogens and for the different kinetics of contact activation reactions exhibited by the two HMW kininogens. PMID- 2989294 TI - Structural organization of the human kininogen gene and a model for its evolution. AB - The entire human kininogen gene has been isolated as a set of overlapping genomic DNA fragments, and the 11 exons encompassing approximately 27 kilobase pairs have been mapped by restriction enzyme analysis and nucleotide sequence determination. The nine 5'-terminal exons encode the 5'-untranslated region and the protein coding region for the signal peptide and the heavy chain, which are common for high molecular weight (HMW) and low molecular weight (LMW) prekininogen mRNAs. Exon 10 consists of the common sequence for bradykinin and the immediately following unique sequence for HMW prekininogen mRNA. Exon 11 is then located following a 90-nucleotide sequence downstream from exon 10 and precisely specifies the sequence unique to LMW prekininogen mRNA. This, together with the hybridization analysis of total human cellular DNA, leads us to conclude that human HMW and LMW prekininogen mRNAs are produced from a single gene as a consequence of alternative RNA processing events. The structural analysis of the kininogen gene also shows that each of the nine 5'-terminal exons discretely specifies the nine protein domains observed in the amino-terminal portion of the kininogens. Furthermore, these nine genetic domains can be characterized by a thrice repeated pattern of three genetic segments, and two sets of these three domains, encompassing exons 3-5 and exons 6-8, are most closely related to each other. Therefore, we have proposed two successive duplication mechanisms as a model for the generation of the structure of the kininogen gene. PMID- 2989296 TI - Dictyostelium amebae alter motility differently in response to increasing versus decreasing temporal gradients of cAMP. AB - Using a perfusion chamber, we examined the behavior of individual amebae in increasing and decreasing temporal gradients of cAMP. We demonstrated that amebae respond to increasing temporal gradients of cAMP with stimulated motility and to corresponding decreasing temporal gradients with depressed motility. Depressed motility observed in decreasing temporal gradients corresponded to the inhibited levels observed when cAMP was applied at constant concentrations. These results were consistent with a simple model for the motile behavior of amebae in an early aggregation territory in which nondissipating waves of cAMP originate at the aggregation center and travel outward periodically. We conclude that chemotactically responsive amebae can assess whether a temporal gradient of chemoattractant is increasing or decreasing in the absence of a spatial gradient, and can adjust their motility accordingly. PMID- 2989295 TI - Sequence rearrangements and genome instability. A possible step in carcinogenesis. AB - A substantial part of the mammalian genome is composed of sequences that do not contain structural genes. These sequences may constitute the major target for physical, chemical and biological DNA-damaging agents and can be involved in carcinogenesis. DNA-damaging agents contribute to the instability of the genome by introducing recombination-prone sites at DNA; these agents lead to extensive chromosomal lesions and rearrangements of genes and their regulatory sequences. Movable sequences that exist and operate in certain bacteria, yeast, and the fruit fly are responsible for sequence rearrangements and contribute to the majority of mutations. Their presence and role in higher animals is not well established. Extensive chromosomal rearrangements were identified in numerous malignancies in man and animals and definitely seem to represent a characteristic of malignancy. Vast chromosomal damage and sequence reshuffling may be of no less importance in the malignant transformation than the point mutation of a particular gene. PMID- 2989297 TI - Effects of chemotactic factors and other agents on the amounts of actin and a 65,000-mol-wt protein associated with the cytoskeleton of rabbit and human neutrophils. AB - Stimulation of rabbit neutrophils by the chemotactic factors fMet-Leu-Phe and leukotriene B4, by platelet activating factor, or by arachidonic acid produces a rapid and dose-dependent increase in the amounts of actin and of a 65,000-mol-wt protein associated with the cytoskeleton. Phorbol 12-myristate, 13-acetate, the calcium ionophore A23187 in the presence or absence of EGTA, and the fluorescent calcium chelator quin-2 also cause an increase in cytoskeletal actin. The stimulated increases in the cytoskeletal actin are not dependent on a rise in the intracellular concentration of free calcium and are not mediated by an increase in the intracellular pH or activation of protein kinase C. The increases in the cytoskeletal actin produced by fMet-Leu-Phe and leukotriene B4, but not by phorbol 12-myristate, 13-acetate, are inhibited by high osmolarity. The effect of hyperosmolarity requires a decrease in cell volume, is not mediated by an increase in basal intracellular concentration of free calcium, and is not prevented by pretreating the cells with amiloride. Preincubation of the cells with hyperosmotic solution also inhibits degranulation produced by all the stimuli tested. The inhibitory action of high osmolarity on the fMet-Leu-Phe and leukotriene B4 induced stimulation of cytoskeletal actin is discussed in terms of the possibility that the addition of high osmolarity, either directly or through activation of protein kinase C, causes receptor uncoupling. PMID- 2989298 TI - An efficient method for introducing macromolecules into living cells. AB - The hemagglutinin (HA) of influenza virus was used to obtain efficient and rapid bulk delivery of antibodies and horseradish peroxidase (HRP) into the cytoplasm of living tissue culture cells. By exploiting HA's efficient cell surface expression, its high affinity for erythrocytes, and its acid-dependent membrane fusion activity, a novel delivery method was developed. The approach is unique in that the mediator of both binding and fusion (the HA) is present on the surfaces of the target cells. A recently developed 3T3 cell line which permanently expresses HA, Madin-Darby canine kidney cells infected with influenza virus, and CV-1 cells infected with a simian virus 40 vector carrying the HA gene were used as recipient cells. Protein-loaded erythrocytes were bound to the HA on the cell surface and a brief drop in pH to 5.0 was used to trigger HA's fusion activity and hence delivery. About 3 to 8 erythrocytes fused per 3T3 and CV-1 cell, respectively, and 75-95% of the cells received IgG or HRP. Quantitative analysis showed that 1.8 X 10(8) molecules of HRP and 1.4 X 10(7) IgG molecules were delivered per CV-1 cell and 6.2 X 10(7) HRP molecules per 3T3 cell. Cell viability, as judged by methionine incorporation into protein and cell growth and division, was not impaired. Electron and fluorescence microscopy showed that the fused erythrocyte membranes remained as discrete domains in the cell's plasma membrane. The method is simple, reliable, and nonlytic. The ability to simultaneously and rapidly deliver impermeable substances into large numbers of cells will permit biochemical analysis of the fate and effect of a variety of delivered molecules. PMID- 2989299 TI - Calcium-dependent association of a protein complex with the lymphocyte plasma membrane: probable identity with calmodulin-calcineurin. AB - A protein complex is shown to participate in a calcium-dependent association with plasma membranes purified either from pig mesenteric lymph node lymphocytes or from human lymphoblastoid cell lines. Plasma membranes prepared in the presence of calcium possess this complex; those prepared in the absence of calcium (5 mM EGTA) do not. The complex associates itself with the inner cytoplasmic surface of the plasma membrane. This complex is referred to as the "acidic protein band" because of its location during migration upon alkaline-urea gel electrophoresis. The complex dissociates from the plasma membrane during electrophoresis on 8-M urea gels, irrespective of calcium levels during electrophoresis; at intermediate urea concentrations (4-6 M), the complex is not dissociated in the presence of calcium. Upon purification of the acidic protein band, SDS acrylamide gel electrophoresis, immunoblotting, and radioimmunoassay techniques suggest that the acidic protein band is composed of at least four peptides (designated 68K, 59K, 20K, 20K): two of these (68K, 20K) are immunopositive for calcineurin and one (20K) is immunopositive for calmodulin. Immunoblots of urea gels also indicate that the calcineurin heavy chain (68K) can also appear at three different locations on the urea gel. Patches and caps induced in human peripheral blood lymphocytes by fluorescein-conjugated goat anti-human IgG are not coincident with the location of calcineurin, which remains distributed throughout the cell. PMID- 2989300 TI - Osmotic and phorbol ester-induced activation of Na+/H+ exchange: possible role of protein phosphorylation in lymphocyte volume regulation. AB - The Na+/H+ antiport is stimulated by 12-O-tetradecanoylphorbol-13, acetate (TPA) and other phorbol esters in rat thymic lymphocytes. Mediation by protein kinase C is suggested by three findings: (a) 1-oleoyl-2-acetylglycerol also activated the antiport; (b) trifluoperazine, an inhibitor of protein kinase C, blocked the stimulation of Na+/H+ exchange; and (c) activation of countertransport was accompanied by increased phosphorylation of specific membrane proteins. The Na+/H+ antiport is also activated by osmotic cell shrinking. The time course, extent, and reversibility of the osmotically induced and phorbol ester-induced responses are similar. Moreover, the responses are not additive and they are equally susceptible to inhibition by trifluoperazine, N-ethylmaleimide, and ATP depletion. The extensive analogies between the TPA and osmotically induced effects suggested a common underlying mechanism, possibly activation of a protein kinase. It is conceivable that osmotic shrinkage initiates the following sequence of events: stimulation of protein kinase(s) followed by activation of the Na+/H+ antiport, resulting in cytoplasmic alkalinization. The Na+ taken up through the antiport, together with the HCO3- and Cl- accumulated in the cells as a result of the cytoplasmic alkalinization, would be followed by osmotically obliged water. This series of events could underlie the phenomenon of regulatory volume increase. PMID- 2989302 TI - Hypoglycemic coma in a high school dropout. PMID- 2989301 TI - Partial disassembly of peroxisomes. AB - Rat liver peroxisomes were subjected to a variety of procedures intended to partially disassemble or damage them; the effects were analyzed by recentrifugation into sucrose gradients, enzyme analyses, electron microscopy, and SDS PAGE. Freezing and thawing or mild sonication released some matrix proteins and produced apparently intact peroxisomal "ghosts" with crystalloid cores and some fuzzy fibrillar content. Vigorous sonication broke open the peroxisomes but the membranes remained associated with cores and fibrillar and amorphous matrix material. The density of both ghosts and more severely damaged peroxisomes was approximately 1.23. Pyrophosphate (pH 9) treatment solubilized the fibrillar content, yielding ghosts that were empty except for cores. Some matrix proteins such as catalase and thiolase readily leak from peroxisomes. Other proteins were identified that remain in mechanically damaged peroxisomes but are neither core nor membrane proteins because they can be released by pyrophosphate treatment. These constitute a class of poorly soluble matrix proteins that appear to correspond to the fibrillar material observed morphologically. All of the peroxisomal beta-oxidation enzymes are located in the matrix, but they vary greatly in how easily they leak out. Palmitoyl coenzyme A synthetase is in the membrane, based on its co-distribution with the 22 kilodalton integral membrane polypeptide. PMID- 2989303 TI - New pharmacologic approaches to thromboembolic disorders. PMID- 2989304 TI - Analysis of soman and sarin in blood utilizing a sensitive gas chromatography mass spectrometry method. AB - Gas chromatography with electron impact mass spectrometry and selected ion monitoring provided a simple and sensitive method for measuring organophosphorus compounds sarin and the two isomers of soman (isomer I and isomer II) in blood. These compounds were extracted from blood or isotonic saline using a modification of the method developed by Sass et al. Blood was deproteinized with perchloric acid before extraction. The acid-induced degradation of the organophosphorus compounds could be minimized by neutralizing the acid immediately after deproteinizing. In saline and blood, 81% of the extractable soman and 74% of the extractable sarin was recovered with a single extraction. The overall recovery of added organophosphorus was less in blood than in saline because of the binding of organophosphorus to blood constituents, probably various enzymes and proteins. A time-dependent decrease in extractable organophosphorus was found in whole blood but not in saline. Although soman isomer II was degraded in blood faster than soman isomer I, no significant difference in the affinities of these two isomers to acetylcholinesterase was observed. PMID- 2989305 TI - Simultaneous assay of choline kinase and choline oxidase in tissue by high performance cation-exchange chromatography and continuous radioactive detection. PMID- 2989307 TI - Rapid purification of human placental angiotensin I converting enzyme by captopril affinity chromatography. PMID- 2989306 TI - Assay of human erythrocyte pyrimidine and deoxypyrimidine 5'-nucleotidase by isocratic reversed-phase high-performance liquid chromatography. AB - We report a rapid and reproducible assay for activity of human erythrocyte pyrimidine 5'-nucleotidase and deoxypyrimidine 5'-nucleotidase. The nucleotides CMP, UMP, dUMP, dCMP or dTMP are individually incubated 30 min at 37 degrees C with erythrocyte hemolysate and 4 mM magnesium chloride in Tris, pH 7.5. Data are provided for standardization of the reaction with each substrate. Individual nucleoside products are assayed in less than 10 min by reversed-phase high performance liquid chromatography at 280 nm with 0-14% methanol in 0.01 M potassium dihydrogen phosphate. This is the first report of a high-performance liquid chromatographic assay system which allows quantitation of the activity of pyrimidine 5'-nucleotidase isozymes using five individual pyrimidine and deoxypyrimidine nucleotides as the substrates. PMID- 2989308 TI - Quantification of ciclopirox by high-performance liquid chromatography after pre column derivatization. An example of efficient clean-up using silica-bonded cyano phases. PMID- 2989310 TI - Determination of specific cytomegalovirus IgM antibodies using infected air dried cells and isolated nuclei by immunoperoxidase assay. AB - A simple immunoperoxidase assay (IPA), adapted for detection of serum IgM antibodies to cytomegalovirus (CMV) is described. The antigen consisted of CMV infected human embryonic fibroblasts or isolated nuclei. The sera were absorbed with aggregated gamma-globulins prior to testing. Rabbit anti-human IgM peroxidase conjugate was used to detect IgM bound to viral antigen. In parallel the enzyme linked immunosorbent assay (ELISA) technique was used to determine IgG and IgM antibodies to CMV, respectively. All patients with acute CMV infections who were tested had CMV-specific IgM antibodies by IPA, both whole cell and nuclei antigen. The maximal IgM titers were higher by ELISA than by IPA but in 3 of the CMV patients IgM was detected earlier by IPA (with both types of antigens) than by ELISA. In 3 of 5 transplant patients with recurrent CMV infection IgM was demonstrated by immunoperoxidase techniques, while by ELISA IgM was demonstrated in only 2 of them. No cross reactivity with other herpes viruses was observed. The described IPA is simple, rapid and has the potential for widespread use in routine laboratories. PMID- 2989309 TI - Antibody-capture ELISA for detection of immunoglobulin M antibodies in sera from Japanese encephalitis and dengue hemorrhagic fever patients. AB - Immunoglobulin M (IgM) antibody titers in paired sera from 19 encephalitis and 44 dengue hemorrhagic fever (DHF) patients in Thailand and 42 Japanese encephalitis (JE) patients in Japan were measured by the antibody capture ELISA and applied to distinguish JE virus infection from dengue virus infection. Titer distribution and the ratio of the titers against JE and dengue antigens led to the following diagnostic criteria. The specimens can be considered as positive with JE when IgM ELISA titer showed over 200 against JE and 4-fold or more higher than titers against any types of dengue antigens. The specimens can be considered as positive with dengue infection when IgM ELISA titer showed over 200 against one of the 4 types of dengue antigens and 4-fold or more higher than against JE antigen. Based on these criteria, 41 of 42 patients in Japan and 11 of 19 encephalitis patients in Thailand could be diagnosed as having JE virus infection while 2 of 19 encephalitis patients in Thailand and 26 of 44 DHF patients in Thailand could be diagnosed as having dengue virus infections. PMID- 2989311 TI - Purification of biologically active Epstein-Barr virus by affinity chromatography and non-ionic density gradient centrifugation. AB - Epstein-Barr virus was purified by affinity chromatography on ricin agglutinin Sepharose followed by non-ionic density gradient centrifugation on Nycodenz. The purified virus was highly active in three biological assays: stimulation of immunoglobulin synthesis by B lymphocytes, transformation of B lymphocytes, and superinfection of Raji cells, as well as in an indirect immunofluorescent binding assay. Electron microscopy revealed intact viral particles free of contaminating membranous structures. CsCl density gradient analysis of nick-translated DNA showed material only at the expected viral density with no detectable material at the density of cellular DNA. SDS gel electrophoresis of radioiodinated virus revealed the characteristic viral polypeptides. This method produces highly purified, biologically active virions with an overall recovery of about 30%. PMID- 2989312 TI - Radioimmunoassay and enzyme-linked immunoassay of antibodies to the core protein (P24) of human T-lymphotropic virus (HTLV III). AB - Human T-cell lymphotropic viruses designated HTLV III or LAV are considered to represent the causative agent(s) of the acquired immunodeficiency syndrome (AIDS). Individuals who have been infected with these viruses may generally be identified on the basis of a positive serological test for antibodies against the protein components of these viruses. Purified viruses or viral proteins have been utilized for developing such tests. Since AIDS may be transmitted by blood transfusion and by blood products, screening of donors for antibodies to HTLV III/LAV has become a necessity. Such screening may be facilitated by the application of assays based on the use of crude virus-infected tissue culture media avoiding elaborate, expensive and potentially hazardous virus purification steps. Serum specimens were mixed with an appropriate dilution of an HTLV III infected tissue culture-derived fraction, obtained by precipitation with polyethylene glycol 6000 and treatment with Tween 80 and tri-n-butylphosphate (to disrupt virus particles), and incubated with polystyrene beads coated with antibodies to HTLV III/LAV (anti-HTLV III). Subsequently, washed beads were incubated with either 125I- or beta-lactamase-labeled anti-HTLV III. The radioactivity or enzymatic activity associated with the beads was proportionate to the quantity of HTLV III antigen originally added to the beads. The presence of anti-HTLV III in serum specimens resulted in decreased antigen binding and thus in decreased radioactivity or diminished beta-lactamase activity associated with the beads. The test was specific for antibodies to the approximately equal to 24 kDa core protein of HTLV III. The prevalence of these antibodies (given in parentheses) in distinct populations was as follows: random blood donors (0.33%); hemophiliacs (36.4%); random homosexual males (25.1%); homosexual males preselected on the basis of positive markers for infection with hepatitis B virus (50%); and those with persistent lymphadenopathy (70%). PMID- 2989313 TI - Purification method for the avian adeno-associated virus. AB - Avian adeno-associated virus (AAAV) pre-purified by Uvasol extraction, one discontinuous and two CsCl equilibrium density gradients, was still considerably contaminated with avian adenovirus (CELO strain). Four different approaches were investigated in attempts to improve the elimination of the contaminating CELO virus. The contaminations were assayed by double immunodiffusion and indirect immunofluorescence. The immunoprecipitation of CELO virus with antiserum and protein A-Sepharose is the most effective method of obtaining purified AAAV free of CELO virus. PMID- 2989314 TI - Antithyrotropin antibodies in the sera of Graves' disease patients. AB - To determine the presence and potential importance of antiidiotypic antibodies (anti-id) in the immune regulation of Graves' disease, sera from 57 patients with Graves' disease were screened before or during antithyroid therapy by enzyme linked immunoabsorbent assay (ELISA) for presumptive anti-id, as defined by the presence of immunoglobulins (Igs) directed against TSH. The mean optical density, indicating the presence of TSH-binding antibodies, was 0.34 +/- 0.28 (+/- SD) in the sera of Graves' disease patients and 0.19 +/- 0.12 in the sera of 24 normal subjects (P less than 0.004). Control antigens (hCG and albumin) did not bind significant amounts of serum Igs. In 8 Graves' patients whose sera bound TSH, 40 80% inhibition was obtained with the addition of TSH receptor-purified IgG (approximately 1 microgram/ml) derived from a single Graves' patient's serum; no inhibition was found with normal IgG (approximately 10 micrograms/ml). Presumptive anti-id was isolated from sera of 6 Graves' patients by affinity purification with a TSH affinity column; the resultant IgG blocked immunoglobulin binding to the TSH receptor when added to the serum of the same patient from whom it had been isolated. The presence of anti-id correlated inversely with the presence of TSH receptor antibodies (r = -0.76; P less than 0.01). These studies demonstrate that 1) significant TSH binding is present in sera from Graves' disease patients, and 2) this TSH binding is specifically inhibitable by Graves' IgG, but not by normal IgG. These data support the hypothesis that TSH-binding immunoglobulins may represent anti-id that are present in Graves' disease as part of the immunological response to TSH receptor or TSH receptor antibodies. Such anti-id could modulate the expression of disease activity in Graves' disease by altering TSH receptor antibody action or production. PMID- 2989315 TI - Augmentation by propranolol of growth hormone-releasing hormone-(1-44)-NH2 induced growth hormone release in normal short and normal children. AB - The effect of a 90-min iv infusion of propranolol, a beta-adrenergic antagonist (0.2 mg/kg BW), on basal plasma GH levels and the GH responses to an iv bolus injection of GH-releasing hormone-(1-44)-NH2 (GHRH; 1 microgram/kg BW) was examined in 10 prepubertal children (6 short but otherwise normal and 4 normal). The iv injection of GHRH resulted in significant increases in plasma GH, comparable to those after either insulin-induced hypoglycemia or arginine infusion. Only a small and inconsistent increase in plasma GH levels occurred during the iv infusion of propranolol, whereas simultaneous administration of propranolol with GHRH caused marked enhancement of GHRH-induced GH release in all subjects. The difference between the plasma GH response to GHRH given with propranolol and the response to GHRH given with 0.9% saline was significantly greater than that between the plasma GH level after propranolol and that after 0.9% saline infusion without GHRH injections. There was no difference in plasma GH responses to GHRH, propranolol, or both in the normal short children or normal children. These findings indicate that beta-adrenergic blockade potentiates GHRH induced GH secretion in prepubertal children. PMID- 2989316 TI - Diurnal variation in the response of plasma adrenocorticotropin and cortisol to intravenous ovine corticotropin-releasing hormone. AB - To determine whether the plasma immunoreactive ACTH (IR-ACTH) and IR-cortisol responses to ovine corticotropin-releasing hormone (oCRH) depend on the time of day, we administered 1 microgram/kg BW synthetic oCRH as an iv bolus dose to five normal men at their usual time of awakening between 0530-0740 h, at 1600 h, and at 2300 h. Mean basal plasma IR-ACTH and IR-cortisol levels were highest upon awakening, intermediate at 1600 h, and lowest at 2300 h, reflecting the diurnal rhythm of ACTH secretion. There was no significant difference in the plasma IR ACTH response to oCRH at different times of the day. In contrast, the mean maximum plasma IR-cortisol increment and mean integrated response were 2- and 2.6 fold greater (P less than 0.05), respectively, at 2300 h than upon awakening. In another study, oCRH was given in the morning (0700-0900 h) to 22 normal men and in the late afternoon (1600-1800 h) to 24 normal men. Mean basal plasma IR-ACTH and IR-cortisol levels were significantly higher (P less than 0.001) in the morning [24 +/- 3 pg/ml (mean +/- SEM) and 10.6 +/- 0.8 micrograms/dl, respectively] than in the afternoon (13 +/- 2 pg/ml and 5.6 +/- 0.6 micrograms/dl, respectively). Mean peak plasma IR-ACTH was slightly greater in the morning (60 +/- 5.5 pg/ml) than in the afternoon (47 +/- 5.5 pg/ml), the mean maximum plasma IR-ACTH increments were the same (35 +/- 4 and 34 +/- 5 pg/ml, respectively), and the mean integrated IR-ACTH response was slightly less in the morning (2036 +/- 414 vs. 2365 +/- 358 pg . min/ml), but none of these differences was statistically significant. Mean peak plasma IR-cortisol concentrations in the morning and afternoon were similar (18.7 +/- 0.7 and 17.3 +/- 0.9 micrograms/dl, respectively), but the mean maximum plasma IR-cortisol increments (8.1 +/- 0.8 and 11.7 +/- 0.9 micrograms/dl, respectively; P less than 0.005), and the mean integrated IR-cortisol responses (588 +/- 115 and 976 +/- 95 micrograms . min/dl, respectively; P less than 0.01) were greater in the afternoon. There was an inverse correlation between basal plasma IR-cortisol concentration and the integrated IR-ACTH response (P less than 0.05), the maximum IR-cortisol increment (P less than 0.001), and the integrated IR-cortisol response (P less than 0.001).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2989317 TI - Dissociation between the effects of endogenous parathyroid hormone on adenosine 3',5'-monophosphate generation and phosphate reabsorption in hypocalcemia due to vitamin D depletion: an acquired disorder resembling pseudohypoparathyroidism type II. AB - In 6 of 8 adults with severe hypocalcemia and osteomalacia due to vitamin D depletion, basal excretion of nephrogenous cAMP (NcAMP) was increased, but the mean renal phosphate threshold (TmP/GFR) was normal, indicating that the steady state phosphaturic response to cAMP generated by endogenous PTH was impaired, as in pseudohypoparathyroidism type II. In all 6 patients, correction of hypocalcemia by administration of vitamin D and calcium restored the normal relationship between NcAMP and TmP/GFR. By contrast, in 13 patients with normocalcemic osteomalacia due to vitamin D depletion, TmP/GFR was reduced, with a significant negative regression on NcAMP, and rose to normal after treatment. Bone histomorphometry after double tetracycline labeling did not differ significantly between the 2 groups. In 72 patients with primary hyperparathyroidism, the slope of the negative regression of TmP/GFR on NcAMP was the same as in normocalcemic secondary hyperparathyroidism, but the adjusted mean for TmP/GFR was significantly lower. We conclude that the effect of endogenous PTH on phosphate reabsorption varies with the level of plasma calcium, and that dissociation between this effect and the generation of cAMP is nonspecific and can be a consequence of hypocalcemia. Exclusion of vitamin D depletion should be an additional diagnostic criterion for pseudohypoparathyroidism type II. PMID- 2989318 TI - Angiotensin II induces the release of 19-hydroxyandrostenedione in man. AB - Plasma concentrations of 19-hydroxyandrostenedione [19-hydroxy-4-androstene-3,17 dione (19-OH-A-dione)], which we reported to be an amplifier of the action of aldosterone on the basis of results obtained in bioassays using adrenalectomized rats, were measured in man using a specific and sensitive RIA. Plasma 19-OH-A dione concentrations (mean +/- SE) in normal subjects were 56 +/- 4 pg/ml in men and 51 +/- 4 pg/ml in women. Plasma 19-OH-A-dione rose significantly during ACTH stimulation and declined significantly during dexamethasone suppression. Plasma 19-OH-A-dione levels in the adrenal vein were an order of magnitude higher than those in the inferior vena cava. These results demonstrate that 19-OH-A-dione is directly secreted by the adrenal cortex and that its secretion is under the control of ACTH. As the action of 19-OH-A-dione may be closely related to that of aldosterone, the response of 19-OH-A-dione to angiotensin II infusion was evaluated and compared with that of aldosterone. During the infusion of graded doses (0.5-4.0 ng/kg . min) of angiotensin II, plasma 19-OH-A-dione increased significantly after the infusion of angiotension II at rates of 1.0 and 2.0 ng/kg . min. It then declined after the infusion of angiotensin II at a rate of 4.0 ng/kg . min. In contrast, plasma aldosterone did not increase significantly until the infusion rate reached 4.0 ng/kg . min. These results indicate that the secretion of 19-OH-A-dione is under the control of angiotensin II. Similar changes in 19-OH-A-dione and aldosterone were found during postural changes. In those subjects who had a small increase in plasma aldosterone when they were upright, 19-OH-A-dione significantly increased. In contrast, in those subjects who had a large increase in plasma aldosterone when upright, 19-OH-A-dione significantly declined. These results suggest that a small elevation of endogenous angiotensin II induces an elevation of plasma 19-OH-A-dione, and a sharp increase in endogenous angiotensin II induces an increase in plasma aldosterone and a decline in plasma 19-OH-A-dione. As angiotensin II stimulated the release of 19-OH-A-dione when the secretion of ACTH was suppressed, angiotensin II acted directly on the adrenal cortex. 19-OH-A-dione is a newly recognized biologically active adrenal C19 steroid which is regulated by both ACTH and the renin-angiotensin system in man. PMID- 2989319 TI - Excess mineralocorticoid receptor activity in patients with dexamethasone suppressible hyperaldosteronism is under adrenocorticotropin control. AB - Total mineralocorticoid activity in human serum was assessed by the rat renal slice receptor assay (RRA). RRA values were compared to RIA-derived aldosterone (aldo) equivalents. Our data demonstrate that in normal subjects, mineralocorticoid receptor-binding steroids can be almost totally accounted for by immunoreactive deoxycorticosterone, corticosterone, cortisol, and aldo (RRA, 4.73 +/- 1.34 ng/ml aldo; RIA, 3.91 +/- 1.52 ng/ml aldo equivalents), while in 8 patients with dexamethasone-suppressible hyperaldosteronism (DSH), RRA values were greater than RIA values in the basal state (RRA, 7.57 +/- 0.75; RIA, 3.24 +/ 0.34; P less than 0.01). DSH patients had a RRA to RIA ratio after ACTH stimulation similar to the ratio in these patients in the basal state (basal, 2.34; ACTH-stimulated, 2.04). During dexamethasone treatment, RRA values fell markedly (1.82 +/- 0.26). Thus, total mineralocorticoid activity, as measured by RRA in DSH patients, was greater than RIA-measurable deoxycorticosterone, corticosterone, cortisol, and aldo, indicating the presence of an unidentified steroid which is dexamethasone suppressible and ACTH stimulable. PMID- 2989320 TI - The inhibitory adenylate cyclase coupling protein in pseudohypoparathyroidism. AB - Deficient activity of the adenylate cyclase stimulatory coupling protein (Ns) has been demonstrated in many patients with pseudohypoparathyroidism type I (PHP) who have Albright's hereditary osteodystrophy and multiple hormone resistance. Since an abnormality in the activity of the related adenylate cyclase inhibitory coupling protein (Ni) could influence hormone responsiveness, we measured pertussis toxin-catalyzed [32P]ADP ribosylation of the 40,000-dalton alpha subunit of Ni (Ni alpha) in erythrocyte membranes from patients with PHP and normal subjects. There were no significant differences in the amounts of Ni alpha in membranes from normal subjects, patients with PHP who have low Ns associated with Albright's hereditary osteodystrophy and multiple hormone resistance, and patients with PHP who have normal Ns. Abnormal Ni is not likely to cause hormone resistance in patients with PHP who have normal Ns or to influence hormone responsiveness in patients with PHP who have low Ns. PMID- 2989321 TI - gamma 3-Melanocyte-stimulating hormone immunoreactivity is a component of the neuroendocrine response to maximal stress (cardiac arrest). AB - The levels of gamma 3-MSH immunoreactivity (gamma 3MSH-IR), beta-endorphin immunoreactivity (beta-endorphin-IR), ACTH, PRL, and cortisol were determined in 23 patients with cardiac arrest (CA) and in a group of 22 patients consecutively admitted to the Intensive Care Unit (ICU controls). Blood was obtained immediately after CA and at frequent intervals for the next 2 h. In ICU patients, blood was obtained the morning after admission. gamma 3MSH-IR was consistently elevated after CA; it was present in all 23 patients and was detectable in 113 of 114 samples. The mean peak gamma 3MSH-IR level was 162 +/- 20 (+/- SE) pg/ml in CA patients and the mean gamma 3MSH-IR level was 35 +/- 6 pg/ml in ICU controls (P less than 0.01). gamma 3MSH-IR was undetectable (less than 20 pg/ml) in normal subjects. CA also was associated with increases in ACTH, beta-endorphin-IR, cortisol, and PRL. The group of ICU controls had stress hormone levels that were generally within the normal range. Distribution of plasma beta-endorphin-IR by gel chromatography showed two peaks of immunoreactivity corresponding to the beta endorphin and beta-lipotropin standards in both the CA and ICU control groups. Distribution of gamma 3MSH-IR in CA plasma showed a major peak of immunoreactivity with a mol wt of approximately 6,000 daltons and two minor components of approximately 4,000 and 11,000 dalton. No immunoreactivity coeluted with a gamma 3MSH standard. We conclude that gamma 3MSH-immunoreactive peptides are a consistent component of the massive release of pituitary and adrenal glandular products after CA. PMID- 2989322 TI - Normal free fatty acid response to isoproterenol in pseudohypoparathyroidism. AB - The lipolytic response to isoproterenol infusion was examined in seven normal subjects and six patients with pseudohypoparathyroidism type I [two had deficiency of the hormone receptor-cyclase coupling protein (N-protein) and four did not]. Despite blunted plasma cAMP responses to isoproterenol in both subgroups of pseudohypoparathyroidism patients, the serum FFA responses were normal. We conclude that changes in plasma cAMP do not reflect the adequacy of the lipolytic response to isoproterenol. PMID- 2989323 TI - Factitious Cushing's syndrome. AB - A woman with pituitary-dependent Cushing's disease remained hypercortisolemic after bilateral adrenalectomy. A search for an adrenal remnant by venous catheterization study suggested persistent cortisol-secreting tissue in the left adrenal bed. During ACTH stimulation, plasma corticosterone concentrations remained low and cortisol remained high, suggesting the cortisol was of exogenous origin. Cushingoid features resolved after confronting the patient with this evidence. Plasma corticosterone concentrations played a novel role in the diagnosis of factitiously induced Cushing's syndrome in this patient. PMID- 2989324 TI - Varicella pneumonia in a bone marrow-transplanted, immune-reconstituted adenosine deaminase-deficient patient with severe combined immunodeficiency disease. AB - Bone marrow transplantation provides an important modality for "enzyme replacement" and the immune reconstitution of patients with adenosine deaminase (ADA) deficiency and severe combined immunodeficiency disease. We report a patient with ADA deficiency who develops severe varicella pneumonia 6 years after successful bone marrow transplantation and immune reconstitution. Marked abnormalities in T-cell mitogen responsiveness and pokeweed mitogen-induced polyclonal immunoglobulin synthesis occurred. Coculture experiments suggested the presence of increased suppressor activity. T-cell phenotyping showed decreased T3 and T4 subsets. These abnormalities slowly resolved over several months as the patient recovered from the varicella infection. ADA enzyme levels and metabolite concentrations in urine and erythrocytes remained unchanged. These findings, together with the chromosome and immune studies, suggested that the bone marrow graft remained intact. These studies indicate that immunologically reconstituted ADA-deficient patients may be at higher risk for complications related to varicella infection and suggest that the institution of preventive measures is important. PMID- 2989325 TI - Enzyme immunoassay versus plaque neutralization and other methods for determination of immune status to measles and varicella-zoster viruses and versus complement fixation for serodiagnosis of infections with those viruses. AB - Results by an enzyme immunoassay method (EIA) performed at one serum dilution and results by indirect immunofluorescence (IFA) and hemagglutination inhibition (HI) tests performed at step dilutions were correlated with results by a neutralization test (50% plaque neutralization [PN]) performed at step dilutions on single serum samples for serologic evaluation of immunity status to measles virus. PN results were taken as true indicators of immunity, and the other tests were evaluated on that basis. The predictive value of a positive result being positive also by PN was 95.3% for HI and 93.3% for EIA and IFA. The predictive value of a negative result being negative also by PN was 81.1% for HI, 100% for EIA, and 75.0% for IFA. A similar study on immunity status to varicella-zoster virus by EIA and by an anticomplement immunofluorescence test versus PN showed a 100% predictive value of a positive or negative result by EIA. By the anticomplement immunofluorescence test, the predictive value of a positive result was 97.7%, and that of a negative result was 88.5%. Studies on the comparative ability of EIA versus complement fixation (CF) to detect significant changes in antibody concentration between acute-phase and convalescent-phase serum samples indicative of a current infection were also done. Both tests were satisfactory for the serodiagnosis of measles or varicella-zoster virus infections. However, EIA was preferable to CF because it was less technically difficult, less labor intensive, and could be performed on sera that were anticomplementary in CF reactions. PMID- 2989326 TI - Immunoglobulin M antibodies detected by enzyme-linked immunosorbent assay and radioimmunoassay in the diagnosis of cytomegalovirus infections in pregnant women and newborn infants. AB - Immunoglobulin M (IgM) antibodies were detected by a commercially available enzyme-Linked Immunosorbent Assay (ELISA) in 36 of 49 (73%) pregnant women with primary cytomegalovirus (CMV) infection. A positive ELISA-IgM result occurred in 10 of 13 patients (77%) assessed within 8 weeks of seroconversion. The sensitivity of the radioimmunoassay (RIA) to identify primary infection in pregnant women was comparable, 78% in general and 86% for women tested within 16 weeks of seroconversion. Of the 36 women with primary infection who had detectable IgM antibodies by ELISA, 25 (69%) were delivered of congenitally infected infants, whereas of the 13 with undetectable IgM antibodies, 7 (54%) transmitted the infection in utero. IgM antibodies were detected by ELISA in only 5 of 43 (11%) women who experienced a recurrence of CMV which either did or did not result in congenital infection. RIA was less likely to measure CMV-specific IgM in recurrent infection, inasmuch as 1 of 19 (5.2%) women with proven recurrent infection had detectable IgM antibody, giving RIA a better specificity for primary infection. Specific IgM antibodies were detected by ELISA in 42 of 61 (69%) babies congenitally infected with CMV and in 4 of 70 (5.7%) uninfected control newborn infants. The RIA was superior for diagnosis of congenital CMV infection, with a sensitivity of 89% and a specificity of 100%. The lower sensitivity of the ELISA-IgM occurred in the category of congenitally infected infants born to mothers with recurrent infection (43%), a group that is at the lowest risk of disease or to develop sequelae. This commercially available ELISA IgM could be used in combination with a CMV-specific IgG test for monitoring women during pregnancy for primary infection. PMID- 2989327 TI - Rapid neutralization assay for human cytomegalovirus antibody. AB - Human cytomegalovirus induces the appearance of immediate early antigens in infected cells 1 h after infection. This provided the basis for the development of a rapid neutralization assay for cytomegalovirus antibody which was able to yield results within a single day. Indirect immunofluorescence to visualize immediate early antigen-positive cells was applied to the rapid determination of cytomegalovirus-neutralizing antibody. The neutralization titers obtained with this assay on 92 serum samples were in accordance with the immune status as determined by enzyme-linked immunosorbent assay to cytomegalovirus-induced immediate early, early, and late antigens. PMID- 2989328 TI - Regulation of glial fibrillary acidic protein (GFAP) expression in CNS development and in pathological states. AB - Studies that have described changes in glial fibrillary acidic protein (GFAP) expression during CNS development and during pathological reactions are summarized. We describe our own studies examining GFAP expression in vitro. In primary cultures established from newborn rat forebrain and maintained in the presence of serum, GFAP expression is first observed in cells from the subventricular germinal zone. Glial precursor cells, at an uncommitted stage, can be induced by serum to begin GFAP synthesis. In astrocytes already established in culture, GFAP synthesis can be further increased by dibutyryl cyclic AMP (dbcAMP). By labelling astrocytes with radioactive methionine, we demonstrate increases of GFAP and vimentin synthesis within 48 h of exposure to dbcAMP. Continuous exposure results in a gradual rise in the cellular level of GFAP. PMID- 2989329 TI - Anaphylatoxin C5a modulation of an alpha-adrenergic receptor system in the rat hypothalamus. AB - C5a anaphylatoxin injected via implanted cannulae into the perifornical region of the hypothalamus stimulated eating in sated rats. C5a also attenuated carbamyl choline-induced drinking, and carbamyl choline inhibited C5a-induced eating, a mutual inhibition characteristic of the adrenergic-cholinergic interactions at this site. The increased food intake induced by C5a was also reversed by phentolamine, an alpha-adrenergic antagonist. Granulocytes infiltrating as a result of C5a-mediated leukotaxis did not arrive at the site in time to influence C5a activity. We propose that C5a in some way activates an alpha-adrenergic receptor system in the hypothalamus, and that anaphylatoxins could mediate neuropsychiatric symptoms sometimes associated with immune complex diseases affecting the central nervous system. PMID- 2989330 TI - Regulation of alpha 1 proteinase inhibitor function by rabbit alveolar macrophages. Evidence for proteolytic rather than oxidative inactivation. AB - Rabbit alveolar macrophages were cultured in an environment conducive to the secretion of both reactive oxygen and proteinases, so that the relative importance of proteolytic and oxidative inactivation of alpha 1-proteinase inhibitor by alveolar macrophages could be evaluated. The inactivation of alpha 1 proteinase inhibitor was proportional to its proteolysis, and there was no detectable inactivation in the absence of proteolysis. Although the live macrophages were capable of secreting reactive oxygen, they did not inactivate alpha 1-proteinase inhibitor by oxidation. The inactivation of alpha 1-proteinase inhibitor by proteolysis was proportional to the secretion of elastinolytic activity by the alveolar macrophages. The inability of the alveolar macrophages to oxidize alpha 1-proteinase inhibitor was attributed to the methionine in the macrophages, in secreted proteins, and in the culture medium competing for oxidants. The data suggest that proteolytic inactivation of alpha 1-proteinase inhibitor may be important in vivo and that the methionine concentration in vivo may protect alpha 1-proteinase inhibitor from significant oxidative inactivation. PMID- 2989331 TI - Continuous administration of synthetic ovine corticotropin-releasing factor in man. Physiological and pathophysiological implications. AB - The continuous 24-h infusion of a maximally stimulating dose (1 micrograms/kg per h) of ovine corticotropin-releasing factor (CRF) in man caused a modest elevation of plasma cortisol (17.2 +/- 1.4 micrograms/dl) and urinary-free cortisol (173 +/ 43 micrograms/24 h) concentrations, which was far less than that seen with a maximally stimulating dose of ACTH (50.4 +/- 2.2 micrograms/dl and 1,200 +/- 94 micrograms/24 h, respectively). The circadian rhythms of plasma ACTH and cortisol were preserved during CRF administration. An intravenous bolus injection of 1 microgram/kg of ovine CRF given to normal volunteers under basal conditions resulted in elevated plasma ACTH and cortisol peak levels (28 +/- 6 pg/ml and 15.0 +/- 1.0 micrograms/dl, respectively). However, no plasma ACTH and cortisol responses were observed when an identical CRF stimulation test was given at the end of the continuous infusion. These findings suggest that the stimulatory activity of exogenous CRF on the ACTH-secreting cells of the pituitary gland is restrained by the negative feedback of cortisol. The persistent circadian rhythm of ACTH, despite a constant level of plasma CRF during the infusion, suggests that the circadian variation in the activity of the hypothalamic-pituitary adrenal axis cannot be explained solely by circadian periodicity of the endogenous CRF stimulus. PMID- 2989332 TI - Characterization of high density lipoprotein binding to human adipocyte plasma membranes. AB - Freshly isolated human adipocytes showed specific uptake of 125I-labeled human high density lipoprotein (HDL2 and HDL3), a portion of which could be released by subsequent incubation with excess unlabeled ligand. To study the mechanism of HDL binding, sucrose gradient-purified adipocyte plasma membranes were incubated with radioiodinated lipoprotein particles under equilibrium conditions in the absence (total binding) or presence (nonspecific binding) of 100-fold excess unlabeled ligand. Specific binding of HDL2 and HDL3, calculated by subtracting nonspecific from total binding, was Ca++ independent, unaffected by EDTA, and not abolished by pronase treatment of the membranes. Modification of HDL3 by reductive methylation or cyclohexanedione treatment also failed to affect its binding to adipocyte plasma membranes. High salt concentration (200 mM NaCl) inhibited specific binding of HDL2 and HDL3 but had no effect on LDL binding. A significant portion of 125I-HDL2 or 125I-HDL3 binding was consistently inhibited by adding excess unlabeled LDL, but this inhibition was incomplete as compared with a similar molar excess of unlabeled HDL2 or HDL3. The role of apoproteins (apo) in HDL binding to adipocyte membranes was examined by comparing binding of HDL2 and HDL3 isolated from normal, abetalipoproteinemic (abeta) and apo E-deficient (apo E0) plasma. Specific binding was observed with all normal and mutant HDL particles. Furthermore, a significant portion (61-78%) of abeta-HDL2, apo E0 HDL2, and apo E0-HDL3 binding was inhibited by adding 100-fold excess of unlabeled low density lipoproteins (LDL). The cross-competition of LDL and HDL binding was confirmed by the ability of normal, abeta, and apo E0-HDL2 to completely inhibit 125I-LDL binding. These data suggest that HDL binding is independent of apo E and that the responsible apoprotein(s) of HDL complete with LDL-apo B for binding to the same or closely related site in the adipocyte plasma membrane. Normal and apo E0-HDL3 binding was also completely inhibited by normal HDL2, which suggested that HDL2 and HDL3 probably bind to the same site. Scatchard analysis of normal HDL2, normal HDL3, and apo E0-HDL3 binding data best fitted a one-component binding profile with similar equilibrium dissociation constants (40-96 nM). HDL3 binding was found to be effectively inhibited by anti human apo AI or anti-human apo AII, but not by anti-human apo B antisera. This binding was also unaffected by monoclonal anti-human apo B or E antibodies known to inhibit binding of apo B or apo E containing lipoprotein to the LDL receptor of cultured fibroblasts. These findings, taken together, suggest that human fat cells possess HDL binding sites with apo AI and /or apo AII specificity. The significant but partial inhibition of HDL2 and HDL3 binding by LDL along with the complete inhibition of LDL binding by HDL2 and HDL3 tends to exclude a single binding site that interacts both lipoproteins and favors the interpretation that LDL and HDL particles bind to multiple recognition sites or to different conformation of the same lipoprotein binding domain on the human fat cell. PMID- 2989333 TI - Race and sex differences in erythrocyte Na+, K+, and Na+-K+-adenosine triphosphatase. AB - Several reports indicate that erythrocytes (RBCs) from blacks and men have higher sodium concentrations than those from whites and women. One possible mechanism to explain this finding is a difference in the activity of Na+-K+-ATPase. To explore this possibility, we have studied the Na+ and K+ kinetics of RBC Na+-K+-ATPase and RBC Na+ and K+ concentrations in 37 normotensive blacks and whites, both males and females. The maximal initial reaction velocity (Vmax) values for RBC Na+-K+-ATPase were lower in blacks and men as compared with whites and women. Higher RBC Na+ levels were observed in blacks and males vs. whites and females. Significant inverse correlations were noted between the Na+-K+-ATPase activity and RBC Na+ concentrations. These findings indicate that cellular Na+ homeostasis is different in blacks and men as compared with whites and women. Since higher RBC Na+ concentrations have also been observed in patients with essential hypertension as compared with normotensive subjects, the higher intracellular Na+ concentrations in blacks and men may contribute to the greater predisposition of these groups to essential hypertension. PMID- 2989334 TI - Deficiency of the autologous mixed lymphocyte reaction in patients with classic hemophilia treated with commercial factor VIII concentrate. Correlation with T cell subset distribution, antibodies to lymphadenopathy-associated or human T lymphotropic virus, and analysis of the cellular basis of the deficiency. AB - 14 patients with hemophilia were studied for the distribution of T cell subsets, the presence of antibody to lymphadenopathy-associated or human T lymphotropic virus type III (LAV/HTLV-III), and their responsiveness in autologous mixed lymphocyte reactions. In addition, mitogen and alloantigen responsiveness and Interleukin-2 production were investigated. Seven patients were found to have low Leu 3a/Leu 2a (T4/T8) ratios; eight patients had antibody to LAV/HTLV-III; and an additional patient had acquired immunodeficiency syndrome. Responsiveness to mitogens and alloantigens as well as Interleukin-2 production were comparable with those of healthy individuals. However, patients with low ratio, many of whom had antibodies to LAV/HTLV-III, had a highly deficient autologous mixed lymphocyte reaction. This reduced response of T cells to autologous non-T cells could not be corrected by elimination of Leu 2a/T8 cells, which indicated that there was a preferential loss of the Leu 3a cell subset(s) which responded to autologous non-T cells. Thus, these patients have a deficiency of intercellular communication within their immune system. PMID- 2989335 TI - Pathogenesis of nephrogenic diabetes insipidus due to chronic administration of lithium in rats. AB - A polyuric syndrome with nephrogenic diabetes insipidus (NDI) is a frequent consequence of prolonged administration of lithium (Li) salts. Studies in the past, mainly the acute and in vitro experiments, indicated that Li ions can inhibit hydroosmotic effect of [8-arginine]vasopressin (AVP) at the step of cAMP generation in vitro. However, the pathogenesis of the NDI due to chronic oral administration of low therapeutic doses of Li salts is not yet clarified. We conducted a comprehensive study to clarify the mechanism by which Li administered orally for several weeks induces polyuria and NDI in rats. Albino rats consuming a diet which contained Li (60 mmol/kg) for 4 wk developed marked polyuria and polydipsia; at the end of 4 wk the plasma Li was 0.7 +/- 0.09 mM (mean +/- SEM; n = 36). Li-treated rats had a significantly decreased (-33%) tissue osmolality in papilla and greatly reduced cortico-papillary gradient of urea (cortex--43%; medulla--64%; papilla--74%). Plasma urea was significantly (P less than 0.001) lower in Li-treated rats (5.4 +/- 0.2 mM) compared with controls (6.8 +/- 0.3 mM). Medullary collecting tubules (MCT) and papillary collecting ducts (PCD) microdissected from Li-treated animals had higher content of protein than MCT and PCD from the control rats. The cAMP accumulation in response to AVP added in vitro was significantly (delta = -60%) reduced. Also, the cAMP accumulation in MCT and PCD after incubation with forskolin was markedly lower in Li-treated rats. Addition of 0.5 mM 1-methyl,3-isobutyl-xanthine did not restore the cAMP accumulation in response to AVP and forskolin in MCT from Li-treated animals. In collecting tubule segments from polyuric rats with hypothalamic diabetes insipidus (Brattleboro homozygotes) the AVP-dependent cAMP accumulation was not diminished. The activity of adenylate cyclase (AdC) in MCT of Li-treated rats, both the basal and the activity stimulated by AVP, forskolin, or fluoride, was significantly (delta approximately equal to -30%) reduced, while the activity of cAMP phosphodiesterase (cAMP-PDIE) in the same segment showed no significant difference from the controls. Also, the content of ATP in MCT microdissected from Li-treated rats and incubated in vitro did not differ from controls. The rate of [14C]succinate oxidation to 14CO2 in MAL was inhibited (-77%) by 1 mM furosemide, which indicates that this metabolic process is coupled with NaCl cotransport in MAL. The rate of (14)CO(2) production from [14C]succinate in MAL was not significantly different between control and Li-treated rats. In MCT of control rats, the rate of [14C]succinate oxidation was approximately 3 times lower than in MAL. The rate of (14)CO(2) production from [(14)C]succinate in MCT of Li treated rats was significantly (delta +33%) higher than in MCT dissected from control rats. Based on these results, we conclude that at least two factors play an important role in the pathogenesis of NDI consequent to chronic oral administration of Li: (a) decreased ability of MCT and PCD to generate and accumulate cAMP in response to stimulation by AVP; this defect is primarily due to diminished activity of AdC in these tubular segments caused by prolonged exposure to Li; and (b) lower osmolality of renal papillary tissue, due to primarily to depletion of urea, which decreases osmotic driving force for water reabsorption in collecting tubules. On the other hand, NaCI reabsorption in MAL is apparently not affected by chronic Li treatment. PMID- 2989336 TI - Characterization of precursor and secreted forms of human angiotensinogen. AB - To define the basis of the heterogeneity of angiotensinogen, we have characterized the immunoreactivity of high molecular weight (HMW) and low molecular weight (LMW) plasma angiotensinogen, the angiotensinogen precursor synthesized by cell-free translation, and angiotensinogen secreted by human hepatoma (Hep G2) cells. Angiotensinogen precursor synthesized by rabbit reticulocyte lysate primed with RNA prepared from liver or Hep G2 cells was compared with angiotensinogen secreted by Hep G2 cells by using immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). So as to assess the contribution of N-glycosylation of angiotensinogen, Hep G2 cells were incubated in the presence of tunicamycin. Glycosylation of secreted angiotensinogen was further characterized by using chromatography on concanavalin A-Sepharose, digestion with neuraminidase, and treatment with trifluoromethane sulfonic acid. In Sephadex G-200 column chromatography, HMW plasma angiotensinogen eluted just after the column void volume and was clearly separated from LMW angiotensinogen which eluted just before bovine serum albumin. Both HMW and LMW plasma angiotensinogen were shown to bind to monoclonal and polyclonal antibodies raised against pure LMW angiotensinogen. Only one angiotensinogen precursor (mol wt 50,000) was identified by cell-free translation which, after cleavage by renin, was reduced to mol wt 45,600. Angiotensinogen secreted by Hep G2 cells showed electrophoretic heterogeneity (mol wt 53,100-65,400). Tunicamycin-treated Hep G2 cells secreted five discrete forms of angiotensinogen, a predominant form of mol wt 46,200, with other forms (mol wt 46,800, 48,100, 49,200, and 49,600) representing 10% of secreted angiotensinogen. All five forms showed a similar reduction in molecular weight after cleavage by renin. The predominant 46,200-mol wt protein represented nonglycosylated angiotensinogen in that, after cleavage by renin, it had an electrophoretic mobility (mol wt 45,600) identical to the desangiotensin I angiotensinogen resulting from renin cleavage of the angiotensinogen precursor. The other higher molecular weight forms of angiotensinogen secreted by tunicamycin-treated Hep G2 cells were shown to represent O-glycosylated angiotensinogen in that they were reduced to 46,200 mol wt by treatment with trifluoromethane sulfonic acid. Dexamethasone (10(-7) and 10(-6)M) stimulated angiotensinogen secretion by Hep G2 cells two- to fourfold, both in the absence and presence of tunicamycin. However, a small stimulatory effect of mestranol (10(-7) M) was evident only in the presence of tunicamycin. Neither dexamethasone nor mestranol influenced the electrophoretic pattern (SDS-PAGE) of angiotensinogen secreted by Hep G2 cells. However, when incubation media were chromatographed on Sephadex G-200 with subsequent immunoprecipitation of the column fractions, both dexamethasone and mestranol were shown to stimulate the secretion of HMW angiotensinogen (eluting just after the column void volume) which, on SDS-PAGE, migrated in a position identical to LMW angiotensinogen. From these studies, we conclude that all forms of human angiotensinogen are derived from a single precursor. The heterogeneity of secreted angiotensinogen represents differences in posttranslational processing of angiotensinogen. This processing includes both N- and O-glycosylation, and also the formation of HMW complexes (HMW angiotensinogen) through association either with other angiotensinogen molecules or with some other protein(s) whose secretion by hepatocytes is stimulated by glucocorticoids and estrogens. PMID- 2989337 TI - Sera from patients with the acquired immunodeficiency syndrome inhibit production of interleukin-2 by normal lymphocytes. AB - We studied the effects of sera from patients with the acquired immunodeficiency syndrome (AIDS) on interleukin-2 (IL-2) production to help elucidate the mechanism of immunodeficiency. Compared with sera from healthy controls, sera from AIDS patients suppressed phytohemagglutinin (PHA)-induced IL-2 production by normal blood mononuclear cells. Sera from homosexual contacts of AIDS patients and from adults with acute cytomegalovirus infection generally lacked this suppressive activity. The effect of the AIDS sera could not be attributed to absence of a stimulatory or nutritive factor, to inactivation of IL-2, to inhibition of the IL-2 assay, nor to increased turnover of IL-2. The suppressive effect of the sera was not mediated by radiosensitive or T8 antigen-bearing suppressor cells or by increased prostaglandin production or decreased interleukin-1 production. The sera acted directly on the groups of cells that produce IL-2, T cells and large granular lymphocytes; suppression occurred at an early, probably pretranslational, stage. When cells were incubated with AIDS sera and then washed, the suppressive effect persisted. The sera did not cause direct or complement-mediated cytotoxic effects on normal mononuclear cells nor did they suppress PHA-induced interferon production, nor proliferation of T lymphoblasts or lymphocyte lines. The suppressive effect was not mediated by interferon, cortisol, immunoglobulin G or M, or immune complexes. The activity was stable at pH 3, pH 10, and 60 degrees C; inactivated at 100 degrees C; and not ether extractable. Because IL-2 plays a central role in the development of many immune responses, the serum factor(s) that inhibits IL-2 production could contribute significantly to the immunodeficiency of AIDS. PMID- 2989338 TI - Administration of atrial natriuretic factor inhibits sodium-coupled transport in proximal tubules. AB - The newly discovered peptides extracted from cardiac atria, atrial natriuretic factors (ANFs), when administered parenterally cause renal hemodynamic changes and natriuresis. The nephron sites and cellular mechanism accounting for profound increase in Na+ excretion in response to ANFs are not yet clarified. In the present study we investigated whether synthetic ANF peptide alters the reabsorption of Na+ and reabsorption of solutes cotransported with Na+ in the proximal tubules of rats. Synthetic ANF peptide consisting of 26 amino acids, 4 micrograms/kg body wt/h, or vehicle in controls, was infused to surgically thyroparathyroidectomized anesthetized rats. After determination of the fractional excretion (FE) of electrolytes (Na+, K+, Pi, Ca2+, Mg2+, HCO3), the kidneys were removed and luminal brush border membrane vesicles (BBMVs) were prepared from renal cortex. Solute transport was measured in BBMVs by rapid filtration techniques. Infusion of ANF peptide increased FENa, FEPi, and FEHCO3; but FECa, FEK, and FEMg were not changed. The increase in FENa was significantly correlated, on the one hand, with increase of FEPi (r = 0.9, n = 7; P less than 0.01) and with increase of FEHCO3 (r = 0.89, n = 7; P less than 0.01). On the other hand, FENa did not correlate with FEK, FECa, or with FEMg. The Na+ gradient dependent uptake of Pi by BBMVs prepared from renal cortex of rats receiving ANF infusion was significantly (P less than 0.05) decreased (-25%), whereas the Na+ gradient-dependent uptake of L-[3H]proline and of D-[3H]glucose or the diffusional uptake of 22Na+ were not changed. ANF-elicited change in FEPi showed a close inverse correlation with decrease of Na+-dependent Pi uptake by BBMVs isolated from infused rats (r = 0.99, n = 7; P less than 0.001). Direct addition of ANF to BBMVs in vitro did not change the Na+ gradient-dependent Pi uptake. In rats infused with ANF, the rate of amiloride-sensitive Na+-H+ exchange across the brush border membrane (BBM) was significantly (P less than 0.05) decreased ( 40%), whereas the diffusional 22Na+ uptake (0.5 min) and the equilibrium (120 min) uptake of 22Na+ were not changed. The inhibition of Na+-H+ exchange after ANF was likely due to alteration of the BBM antiporter itself, in that the H+ conductance of BBMVs was not increased. We conclude that synthetic ANF (a) decreases tubular Na+ reabsorption linked to reabsorption of HCO3 in proximal tubules, and (b) inhibits proximal tubular reabsorption of Pi coupled to Na+ reabsorption, independent of secretion and/or action of parathyroid hormone or calcitonin. These ANF effects are associated with inhibition of Na+-Pi synport and of Na+-H+ antiport in luminal BBMs. Our findings document that inhibition of Na+-coupled transport processes in proximal tubules is an integral part of the renal response to ANF. PMID- 2989339 TI - Abnormal function of B lymphocytes from peripheral blood of multiple myeloma patients. Lack of correlation between the number of cells potentially able to secrete immunoglobulin M and serum immunoglobulin M levels. AB - Multiple myeloma patients are deficient in normal polyclonal serum immunoglobulins. To determine the reasons for this decrease, we quantitated and compared the number of surface IgM+ B lymphocytes, and the number of B cells susceptible to transformation by Epstein-Barr virus (EBV) with the concentration of IgM in serum. Serum IgM levels varied considerably in individual patients, temporally shifting from undetectable to normal amounts and then dropping again to undetectable levels. A transient rise to normal serum IgM concentrations was seen in 42% of patients assessed at two or more time points. Of 44 patients, 52% showed a lack of correlation between the number of surface IgM+ (sIgM+) B cells and serum IgM concentration. One subset of patients (25%) had detectable to normal numbers of sIgM+ B cells in blood but undetectable levels of serum IgM. Transformation of B cells from these patients indicated a block in IgM secretion that was extrinsic to the B cells that were fully able to transcribe, translate, and secrete IgM after EBV transformation. A second subset of patients (27%) had undetectable numbers of sIgM+ B cells but near normal levels of serum IgM, suggesting abundant secretion by few clones of B cells. Of 18 patients with monoclonal gammopathy of undetermined significance (MGUS), 26% showed a lack of correlation between the numbers of sIgM+ B cells and serum IgM concentration. We suggest that in patients with multiple myeloma, and in some with MGUS, there exists a mechanism(s) extrinsic to the B cell that mediates an arrest in terminal B lymphocyte maturation. PMID- 2989341 TI - Hepatic copper distribution in primary biliary cirrhosis shown by the scanning proton microprobe. AB - A number of conditions are associated with abnormalities of trace metal handling by the liver. We report the application of the Oxford scanning proton microprobe to the analysis of hepatic copper in one such condition, primary biliary cirrhosis. The scanning proton microprobe analyses conventional tissue sections (5-10 micron thickness) and produces simultaneous elemental distribution maps of biologically relevant elements with a spatial resolution of 1 micron and a detection limit better than 1 ppm. We have confirmed the localisation of excess copper to periportal areas and suggest that such accumulation is confined to a proportion of periportal hepatocytes. We have also shown a close spatial correlation between regions of copper accumulation and areas of high sulphur concentration. The copper to sulphur ratio in these areas is consistent with their identity as aggregates of copper loaded metallothionein, and the scanning proton microprobe was further able to show that the aggregates contain less than 30 ppm zinc. PMID- 2989340 TI - Cyclic adenosine monophosphate-stimulated bicarbonate secretion in rabbit cortical collecting tubules. AB - We studied the effects of cyclic AMP (cAMP) on HCO-3 transport by rabbit cortical collecting tubules perfused in vitro. Net HCO-3 secretion was observed in tubules from NaHCO3- loaded rabbits. 8-Bromo-cAMP-stimulated net HCO-3 secretion, whereas secretion fell with time in control tubules. Both isoproterenol and vasopressin (ADH) are known to stimulate adenylate cyclase in this epithelium; however, only isoproterenol stimulated net HCO-3 secretion. The mechanism of cAMP-stimulated HCO-3 secretion was examined. If both HCO-3 and H+ secretion were to occur simultaneously in tubules exhibiting net HCO-3 secretion, cAMP might increase the net HCO-3 secretory rate by inhibiting H+ secretion, by stimulating HCO-3 secretion, or both. These possibilities were examined using basolateral addition of the disulfonic stilbene (4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS). In acidifying tubules from NH4Cl-loaded rabbits, DIDS eliminated HCO-3 reabsorption, a result consistent with known effects of DIDS as an inhibitor of H+ secretion. In contrast, cAMP left acidification (H+ secretion) intact. DIDS applied to HCO-3 secretory tubules failed to increase the HCO-3 secretory rate, indicating minimal H+ secretion in HCO-3 secreting tubules. Thus, inhibition of H+ secretion by cAMP could not account for the cAMP-induced stimulation of net HCO-3 secretion. cAMP-stimulated HCO-3 secretion was reversibly eliminated by 0 Cl perfusate, whereas luminal DIDS had no effect. Bath amiloride (1 mM) failed to eliminate cAMP-stimulated HCO-3 secretion when bath [Na+] was 145 mM or 5 mM. cAMP depolarized the transepithelial voltage. The collected fluid [HCO-3] after cAMP could be accounted for by electrical driving forces, suggesting that cAMP stimulates passive HCO-3 secretion. However, cAMP did not alter HCO-3 permeability measured under conditions expected to inhibit transcellular HCO-3 movement (0 Cl- solutions and bath DIDS). This measured HCO-3 permeability was not high enough to account, by passive diffusion, for the HCO-3 fluxes observed in Cl-containing solutions. We conclude the following: cAMP increased net HCO3- secretion by stimulating HCO3- secretion and not by inhibiting H+ secretion; this HCO3- secretion may have occurred by Cl-HCO3- exchange; Na+-H+ exchange appeared not to play a role in basolateral H+ extrusion under these conditions; and the stimulation of HCO3- secretion by isoproterenol, but not ADH, suggests the existence of separate cell cAMP pools or cellular heterogeneity in this cAMP response. PMID- 2989343 TI - Angiokeratoma corporis diffusum and normal enzyme activities. PMID- 2989342 TI - Detection of rotavirus by a latex agglutination test, Rotalex: comparison with electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. AB - A commercially available latex agglutination test, Rotalex (Orion Diagnostics, Finland), for detecting rotaviruses was evaluated in comparison with four other tests (electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay) routinely used in our laboratories. Although Rotalex was the least complex method, it showed lack of specificity and sensitivity when carried out according to the manufacturer's instructions. Four basic modifications of Rotalex are described. These include the use of Hank's balanced salt solution, increasing the incubation time to 20 min, reading the agglutination result by an experienced observer, and the use of 50 mm square glass plates. The modified procedure gave results which were comparable with those obtained by electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. The latter techniques, when used to detect rotavirus, all gave similar results. PMID- 2989344 TI - Glucagonoma syndrome. A clinicopathologic, immunocytochemical, and ultrastructural study. AB - Necrolytic migratory erythema has been considered to be a paraneoplastic dermatosis. Although some individuals have had no associated neoplasm, a pancreatic glucagonoma is typically found in the vast majority. We describe a 71 year-old woman with findings strongly suggestive of the glucagonoma syndrome. The plasma glucagon level was modestly elevated. Special studies of the pancreas, including computerized axial tomography and selective angiography, did not demonstrate a tumor. Surgery, however, revealed a 5-cm tumor in the head of the pancreas. Although immunocytochemical stains showed only rare, weakly reactive cells, the electron microscopic study confirmed the diagnosis of an islet cell tumor and revealed characteristic morphologic features of glucagonoma. PMID- 2989345 TI - Herpes simplex complicating topical 5-fluorouracil therapy. PMID- 2989346 TI - The effect of soluble pyrophosphates on dental calculus in adults. AB - A sodium fluoride dentifrice, containing a mixture of soluble pyrophosphates, was used ad libitum in an adult population for 6 months after receiving a dental prophylaxis. In comparison with the control group, who used the sodium fluoride formula without the pyrophosphates, the test group had significantly less calculus by occurrence and severity. Thus, a significant reduction in newly forming calculus was obtained with the experimental dentifrice when used as an adjunct to regular professional care and personal oral hygiene procedures. PMID- 2989347 TI - Measurement of myocardial infarction fraction using single photon emission computed tomography. AB - Although infarct size correlates generally with prognosis after acute myocardial infarction, an absolute measure of infarct size may have differing prognostic significance depending on absolute left ventricular mass. To test the hypothesis that single photon emission computed tomography can accurately measure myocardial infarct size as a percent of total left ventricular mass ("infarction fraction"), thallium-201 and technetium-99m pyrophosphate tomograms were acquired in 21 dogs 24 to 48 hours after fixed occlusion of the left anterior descending or circumflex coronary artery. Pathologic infarct weight was measured as the myocardial mass that showed no staining with triphenyltetrazolium chloride. Scintigraphic infarct mass by technetium-99m pyrophosphate was calculated from the total number of left ventricular volume elements (voxels) demonstrating technetium-99m pyrophosphate uptake X voxel dimension [( 0.476 cm]3) X specific gravity of myocardium (1.05 g/cm3). Scintigraphic left ventricular mass was calculated in a similar fashion using an overlay of the thallium-201 and technetium-99m pyrophosphate scans. The "infarction fraction" was calculated as: infarction fraction = infarct mass/left ventricular mass. There was good correlation between single photon emission computed tomography and pathologic measurements of infarct mass (technetium-99m pyrophosphate mass = 1.01 X pathologic infarct mass + 0.96; r = 0.98), left ventricular mass (single photon emission computed tomographic left ventricular mass = 0.60 X pathologic left ventricular mass + 37.4; r = 0.86) and "infarction fraction" (single photon emission computed tomographic infarction fraction = 1.09 X pathologic infarction fraction - 1.7; r = 0.94).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989348 TI - Occurrence and transmission of localized repolarization abnormalities in vitro. AB - Abnormal delay of repolarization leading to prolonged reduction of membrane potential could lead to arrhythmias. Transmission of this type of abnormality from an "abnormal" to a "normal" segment of Purkinje fiber was studied as follows: In a double compartment bath one segment was superfused with unmodified Tyrode's solution and the other with solutions known to induce abnormal prolongation of action potential duration (manifest by prolonged step-like delays in repolarization, or "secondary plateaus" and "early afterdepolarization," that is, activations arising as a consequence of the prolonged depolarized membrane potential). Transmission of abnormalities varied. Complete transmission of secondary plateaus and early afterdepolarizations led to action potentials configurationally similar throughout the fiber. Selective transmission of early afterdepolarization led to the occurrence of new premature action potentials in the normal segment. Cycle length prolongation favored both the occurrence and the transmission of abnormalities. After addition of lidocaine or high potassium ion solutions to the normal segment, shortening of action potential duration occurred and was transmitted from normal to abnormal segment. In this way, reversal of abnormality throughout the fiber occurred via an intervention applied selectively to the normal segment. Observations in this study may be relevant to arrhythmias and their treatment. PMID- 2989349 TI - Acute and long-term effects of enalapril on the cardiovascular response to exercise and exercise tolerance in patients with congestive heart failure. AB - Enalapril is a recently developed angiotensin-converting enzyme inhibitor that improves cardiac function at rest in patients with congestive heart failure. This study investigated the acute effects of enalapril on the cardiovascular response to exercise, and then evaluated the long-term effects of enalapril on exercise capacity and functional status during a 12 week placebo-controlled trial in patients with heart failure. Ten patients underwent hemodynamic monitoring while at rest and during incremental bicycle exercise before and after 5 to 10 mg of enalapril orally. At rest, enalapril decreased mean blood pressure 13% (p less than 0.01) and systemic vascular resistance 20% (p less than 0.05) and increased stroke volume index 21% (p less than 0.01). During maximal exercise, enalapril decreased systemic vascular resistance and increased both cardiac and stroke volume indexes. Enalapril acutely increased exercise duration (p less than 0.05) and maximal oxygen consumption (p less than 0.001). These 10 patients and an additional 13 patients were then randomized to either placebo or enalapril treatment and followed up for 12 weeks. Of the 11 patients assigned to active treatment, 73% considered themselves improved compared with 25% of the patients assigned to placebo treatment (p less than 0.02). During long-term treatment, exercise capacity increased in patients receiving enalapril (p less than 0.001) but was unchanged in patients receiving placebo (intergroup difference, p less than 0.05). During long-term treatment, no adverse effects of enalapril occurred. Thus, enalapril improves cardiac function at rest and during exercise. Compared with placebo, maintenance therapy with enalapril results in symptomatic improvement and increased exercise capacity. PMID- 2989350 TI - The oxygen free radical system: potential mediator of myocardial injury. AB - The sequential univalent reduction of oxygen gives rise to very reactive intermediate products including superoxide anion radical, hydrogen peroxide and free hydroxyl radicals. Normally, the tissue concentration of these intermediate products of oxygen is severely limited; however, if oxygen free radicals are produced in excess of the capacity of the tissues to eliminate them, they may cause serious damage. The biochemistry and possible sources of free radical generation in animal models of ischemic/reperfusion injury are reviewed. The ability of scavengers of oxygen free radicals to improve mechanical, mitochondrial and sarcoplasmic reticulum function in animal models of ischemic/reperfusion injury suggests that oxygen free radicals are partly responsible for myocardial injury in these models. Future research should be directed at establishing the relevance of oxygen radical-mediated myocardial injury in the experimental setting to analogous clinical situations. PMID- 2989351 TI - Subchronic toxicology of butyl nitrites in mice by inhalation. AB - Mice were exposed by inhalation to n-butyl, iso-butyl sec-butyl or tert-butyl nitrite in a dynamic airflow chamber 7 h per day for 60 days at concentrations that caused less than 20% fatalities. Under these conditions, body-weight gain was depressed over the first 30 days by all four compounds, but returned to normal over the final 30 days for all compounds except tert-butyl nitrite. Spleen weights were increased by all four butyl nitrites, and lung weights were increased by all except sec-butyl nitrite. Kidney weights were increased by iso butyl and sec-butyl nitrites, but decreased by the tert-butyl compound. Liver weights were increased by iso-butyl nitrite exposure and decreased by tert-butyl nitrite exposure. The results are consistent with the hypothesis that a significant aspect of butyl nitrite toxicity is due to the resultant methemoglobinemia. PMID- 2989352 TI - [Value of alveolar lavage and transbronchial biopsy in the diagnosis of isolated sarcoid uveitis]. AB - The diagnosis of sarcoidosis is established by biopsy of sarcoid tissues demonstrating non-caseating granuloma, but difficulties arise when extra pulmonary organs are involved separately. Positive histologic signs can however be found in the lung even when there are no radiologic features. Bronchoalveolar lavage and biopsies were performed in 19 patients presenting severe, strictly isolated uveitis (5 cases of anterior uveitis, 3 cases of posterior uveitis and 11 cases of panuveitis). Criteria of positivity were non-caseating granuloma and lymphocytosis. Positive signs were obtained in 6 cases (31,5%); 1 case of posterior uveitis and 5 of panuveitis. There was no case of false positive results but one false negative result. There was a lack of correlation between results of these investigations and angiotensin converting enzyme blood levels. These investigations are non-invasive in experienced hands. PMID- 2989353 TI - Aging: effects on sodium- and potassium-activated adenosine triphosphatase activity and ouabain binding sites in rat brain. AB - Sodium- and potassium-activated adenosine triphosphatase (Na,K-ATPase) activity and [3H]-ouabain binding were examined in homogenates of cerebral cortex, striatum, and hypothalamus of 3-, 8-, and 26-month-old rats to determine if aging related alterations in energy utilization demonstrated in brain slices and homogenates are potentially associated with alterations in Na,K-ATPase. There were no consistent age-related changes seen in Na,K-ATPase activity, the number of [3H]-ouabain binding sites, or their affinity for ouabain. Moreover, enzyme activities of the two molecular forms of Na,K-ATPase and their inhibition by strophanthidin did not appear to be different in partially purified enzyme preparations obtained from whole brain of 3- and 26-month-old rats. In contrast, the concentration of [3H]-ouabain binding sites was lower in cardiac muscle of senescent rats indicating a reduction in the number of active Na,K-ATPase units. It appears unlikely that aging-related central nervous system changes are associated with alterations in the Na,K-ATPase enzyme system. PMID- 2989354 TI - Physical-chemical and biochemical differences in liver plasma membranes in aging F-344 rats. AB - Age-associated differences in the microviscosity and the activities of enzymes (Mg++-ATPase, Na+,K+-ATPase and 5'-nucleotidase) in the liver plasma membrane were investigated in male and female rats of various ages ranging from 2 to 30 months. The membrane microviscosity, as determined by fluorescence polarization using 1,6-diphenyl-1,3,5,-hexatriene (DPH) as a probe, increased progressively with age after 2 months in male rats, whereas in female rats the microviscosity began to increase only after 24 months. On the other hand, age-associated differences in the activities of membrane-bound enzymes were generally minimal or not significant with the exception of the 5'-nucleotidase activity determined at the pH 9.1, which progressively decreased with age in male rats. The Na+, K+ ATPase activity tended to decrease in both sexes. These decrements in activity did not appear to be large enough, however, to be of definitive physiological significance. These results suggest that age may change the physical-chemical and biochemical qualities of the rat's hepatocyte plasma membrane, but the relationship between the membrane microviscosity and the activities of membrane bound enzymes is not a simple parallelism. PMID- 2989355 TI - Thyrotropic hormone and thyroidal function initiation in fetal rat. AB - In rat fetus thyrotropic hormone (TSH) is present in the blood as early as day 16 and reaches maximum on day 18. On day 21, TSH level falls off to an intermediate value. Thyroxine is detected in the plasma of fetuses from day 18; the subsequent increase in T4 concentration until day 21, probably generates the negative feedback controlling TSH secretion. The observed increase and maximum in plasma TSH are concomitant with the highest concentrations of cAMP in the thyroid. There may be a causal relationship behind this concomitance since TSH stimulates in vitro the accumulation of cAMP in explanted thyroid from 17 days-old fetuses. The implications of these findings on the role of TSH in the developing rat thyroid are discussed. PMID- 2989356 TI - The etiology of diarrhoea studied in the community in young urban Gambian children. PMID- 2989357 TI - Computed tomography in the evaluation of cirrhosis and portal hypertension. AB - Patients with cirrhosis present a continuing diagnostic and therapeutic challenge. The status of their disease frequently changes, necessitating intensive serial evaluation. CT is an invaluable tool in the management of these patients because it can noninvasively provide vital information concerning liver size, contour, and occasionally hepatic parenchyma. More importantly, CT can demonstrate superficial and deep varices, assess the patency of the extrahepatic portal system, and detect other complications including ascites, hepatic steatosis, hemochromatosis, and hepatocellular carcinoma. PMID- 2989359 TI - Surveillance of respiratory viral infections by rapid immunofluorescence diagnosis, with emphasis on the epidemiological development of respiratory syncytial virus infections. AB - Surveillance of certain respiratory viral infections by applying immunofluorescence (IF) examinations to samples of nasopharyngeal secretions has been evaluated using a simplified procedure for the preparation of cell smears. Samples from 711 children living in different parts of Norway were examined during the winter 1982/83 and a positive diagnosis was made for 290 children (41%). Temporal epidemic peaks were observed for respiratory syncytial virus (RSV), parainfluenza virus type 3 and influenza virus. On the other hand, the monthly number of negative samples was almost constant throughout the period. Differences in timing of RSV outbreaks were observed between two regions in Norway. Compared to rapid IF diagnosis, RSV notifications obtained by serological examinations were delayed by several weeks. Rapid virus diagnosis by IF examinations with our simplified procedure for preparation of nasopharyngeal samples seems to be suitable for the epidemiological surveillance of respiratory viral infections, both for its simplicity of preparation of the samples and for its accuracy in defining the time of the actual virus infection. Nevertheless, the method is not without pitfalls; a close cooperation between those who take the specimens and the laboratory is essential, and the IF examinations should be performed by an experienced microscopist. PMID- 2989358 TI - A study of enterotoxigenic Escherichia coli, serogroup 0126, by bacterial restriction endonuclease DNA analysis (BRENDA). AB - Sixteen isolates of Escherichia coli were subjected to bacterial restriction endonuclease DNA analysis (BRENDA). Nine of these isolates were from an outbreak of human diarrhoea and produced stable toxin, the remaining seven were non toxigenic strains from animal and human sources. The isolates from the outbreak produced indistinguishable DNA electrophoretic patterns in spite of their assignment to seven different H serotypes. Their BRENDA patterns were markedly different from the other isolates examined. These results support the epidemiological evidence that a single-strain outbreak had occurred, and they cast doubt on the value of H typing for this particular investigation. PMID- 2989360 TI - A preliminary survey of the epidemiology of bluetongue in Costa Rica and northern Colombia. AB - Recent evidence of bluetongue (BT) virus infection of livestock in scattered localities in the neotropics prompted a serologic survey of cattle in Colombia and Costa Rica. In Costa Rica 48.1% of 1435 bovine animals had BT virus antibody in the agar gel precipitation test (AGPT). In Colombia 51.8% of 635 cattle were AGPT-positive for BT virus. Antibody prevalence ranged from over 50% in the lowlands to 0% in Costa Rica and 19% in Colombian cattle above 2000 m altitude. Neutralization tests indicated that Costa Rican cattle had been exposed to BT virus types 6, 12, 14 and 17. PMID- 2989361 TI - Sodium periodate-induced T cell mitogenesis: an analysis of the requirement for Ia and IL 1. AB - T lymphocytes oxidized with the mitogen sodium periodate undergo a proliferative response when cultured in the presence of Ia+ accessory cells. However, the exact role(s) the accessory cells play in such a response has not been clearly defined. We have evaluated the role of Ia and the requirement for interleukin 1 (IL 1) in periodate mitogenicity by using the Ia+ cloned tumor cell lines P388AD (Ia+, IL 1 inducible) and P388NA (Ia+, IL 1 noninducible) as accessory cells. P388AD but not P388NA was able to supply accessory cell function to periodate-treated T cells, suggesting that Ia expression alone was not sufficient to reconstitute a response. Monoclonal anti-I-Ad and anti-I-Ed antibody blocked the accessory cell function of P388AD. In addition, monoclonal antibody GK 1.5, directed against the T cell determinant L3T4a, blocked the P388AD/periodate-treated T cell interaction, confirming that this interaction was restricted by class II molecules. Although Ia expression was required, the response was not major histocompatibility complex (MHC) restricted, because allogeneic as well as syngeneic macrophages were capable of supplying accessory cell function to periodate-treated T cells. Exogenous IL 1 alone was able to trigger periodate treated T cells, suggesting that Ia was required for the induction of IL 1 synthesis by the accessory cells. Furthermore, purified IL 2, devoid of IL 1 activity, was able to fully reconstitute the proliferative response of accessory cell-depleted oxidized T cells to a level equal to that of whole spleen accessory cells or P388AD. These data suggest that periodate-treated T cells can proliferate with IL 1 alone and that Ia+ accessory cells in periodate-mediated T cell mitogenicity may function in the release of IL 1 and the induction of IL 2 synthesis by the T cells. PMID- 2989362 TI - Prostaglandin E2 acts at two distinct pathways of T lymphocyte activation: inhibition of interleukin 2 production and down-regulation of transferrin receptor expression. AB - The mechanism by which prostaglandin E2 (PGE2) inhibits human T lymphocyte activation and proliferation was studied. We analyzed the effect of physiologic concentrations of PGE2 on interleukin 2 (IL 2) production, expression of IL 2 receptor (Tac antigen), and expression of the transferrin receptor after in vitro activation with phytohemagglutinin. PGE2 inhibited T lymphocyte proliferation by 80 to 90% of control values. This was associated with a similar degree of inhibition of IL 2 production while the expression of IL 2 receptor was not affected. This was in marked contrast to the expression of the transferrin receptor, which was inhibited 65% after 72 hr of in vitro activation. The addition of exogenous, purified IL 2 reconstituted lymphocyte proliferation to 50% of control values, but had no effect on transferrin receptor expression. Because PGE2 is known to increase the intracellular concentration of 3',5' cyclic adenosine monophosphate (cAMP), we investigated the effect of another adenylate cyclase activator, i.e., isoproterenol, as well as the effect of extracellular administration of the cAMP derivative dibutyryl cAMP (dBcAMP) on IL 2 production, Tac antigen expression, and transferrin receptor expression. It was demonstrated that isoproterenol, as well as dBcAMP, inhibited transferrin receptor expression on PHA-activated T lymphocytes to the same extent as PGE2, and exogenous IL 2 could not counteract the down-regulation of the receptor expression. In contrast, neither isoproterenol nor dBcAMP had any significant effect on IL 2 receptor expression. Prostaglandin F2 alpha (PGF2 alpha), which has been reported to elevate intracellular cyclic GMP levels, had no effect on lymphocyte activation and proliferation, and did not counteract the PGE2-induced depression in IL 2 production. In contrast to its effect on peripheral blood lymphocytes, PGE2 had no effect on transferrin receptor expression or cell proliferation by IL 2 dependent T cell clones and IL 2-independent T cell lines. These studies demonstrate that PGE2 exerts its inhibitory effects on T cell activation and proliferation via two distinct pathways: inhibition of IL 2 production and inhibition of transferrin receptor expression. The transferrin receptor inhibition is mediated via the cAMP pathway and is IL 2-independent. PMID- 2989363 TI - Thymocyte cyclic AMP and cyclic GMP response to treatment with metabolites issued from the lipoxygenase pathway. AB - Evidence has been presented that cGMP is the second messenger for the lipoxygenase metabolites 15-HETE and LTB4 in the mouse splenocyte and thymocyte. Incubation of splenocytes with 10(-7) to 10(-9) M 15-HETE caused a slight decrease in cAMP levels and an increase in cGMP levels after 10 to 20 min. Mature PNA-, immature PNA+, and whole thymocytes treated with 10(-7) to 10(-10) M 15 HETE and 10(-11) M LTB4 showed an approximately 100% increase in cGMP production. In mixed lymphocyte reactions, 15-HETE- and LTB4-treated PNA+, PNA-, and whole thymocyte populations inhibited thymidine uptake by fresh allostimulated splenocytes. These results demonstrate that the eicosanoid-induced generation of suppressor cells follows a rise in lymphocyte cGMP levels. PMID- 2989364 TI - Adenosine; a physiologic modulator of superoxide anion generation by human neutrophils. Adenosine acts via an A2 receptor on human neutrophils. AB - Adenosine specifically inhibits superoxide anion generation by N-formyl-methionyl leucyl-phenylalanine-stimulated neutrophils without affecting either degranulation or "aggregation." We present data that also supports the hypothesis that adenosine engages a specific cell surface receptor to mediate inhibition of stimulated neutrophils. Theophylline (10 and 100 mu M), a competitive antagonist at adenosine receptors, reversed the effects of adenosine (0.1 mu M) on superoxide anion generation by stimulated neutrophils. The adenosine analogue 5'N ethylcarboxamidoadenosine (NECA) was a more potent inhibitor of superoxide anion generation than either N6-phenylisopropyladenosine (PIA) or adenosine, an order of potency consistent with that previously demonstrated for adenosine A2 receptors. 2-Chloroadenosine inhibited superoxide anion generation at concentrations similar to NECA. [3H]-NECA and [3H]-2-chloroadenosine bound to a single receptor on intact neutrophils. The characteristics of the receptors for [3H]-NECA and [3H]-2-chloroadenosine were similar (Kd = 0.22 and 0.23 mu M, respectively; number of binding sites = 9.31 and 11.1 X 10(3) sites/cell, respectively). NECA, 2-chloroadenosine, adenosine, and PIA inhibited binding of [3H]-NECA with a rank order similar to that for inhibition of superoxide anion generation (NECA = 2-chloroadenosine greater than adenosine greater than PIA). There was 50% inhibition of superoxide anion generation by NECA at approximately 20% receptor occupancy. Adenosine, derived from damaged tissues, may serve as a specific, endogenous modulator of superoxide anion generation by activated neutrophils through interaction at this newly described receptor on human neutrophils. PMID- 2989365 TI - Abrogation of anti-Pichinde virus cytotoxic T cell memory by cyclophosphamide and restoration by coinfection or interleukin 2. AB - Previously, we demonstrated that memory cell-mediated immune responses can be generated in Pichinde virus (PV)-primed mice after secondary challenge in vivo with homologous virus. Further, treatment of mice with cyclophosphamide (CY) before primary infection with PV abrogated the generation of H-2-restricted, virus-specific cytotoxic T lymphocytes (CTL), and rechallenge of these mice was followed by neither a primary nor a secondary CTL response. Here, we demonstrate that this CY-induced block in memory anti-PV CTL generation was not due to establishment of a persistent infection. Interestingly, this CY-induced block in memory anti-PV CTL generation was overcome by secondarily coinfecting mice with PV and lymphocytic choriomeningitis virus (LCMV) or PV and Tacaribe virus. Secondary infection with LCMV or Tacaribe virus alone did not elicit anti-PV CTL. Coinfection resulted in the generation of a PV-specific memory CTL response as judged by maximal activity on day 4 after rechallenge. Co-infection with PV and vesicular stomatitis virus, an unrelated rhabdovirus, did not efficiently restore memory anti-PV CTL responses. Memory anti-PV CTL responses were also restored when interleukin 2 (IL 2)-containing supernatants were injected i.p. after rechallenge of CY-treated mice with PV. To demonstrate that IL 2 was the responsible lymphokine in these preparations, highly purified IL 2 was added to in vitro cultures of spleen cells from CY-treated PV-primed mice. In the presence of PV-infected syngeneic macrophages, addition of purified IL 2 resulted in a dose-dependent restoration of H-2-restricted anti-PV CTL activity. The CTL precursor (CTLp) frequency of CY-treated PV-primed mice was markedly decreased from that of normal PV-primed mice. Thus, the long-lasting block in the ability to generate a PV-specific memory CTL response after CY treatment appears to be due to both a lack of helper T cell activity and a significant reduction of CTLp. However, this block may be overcome by coinfecting with viruses that cross-react at the helper T cell level or by exogenous treatment with highly purified IL 2. PMID- 2989366 TI - Defective cell-mediated immunity in leprosy: failure of T cells from lepromatous leprosy patients to respond to Mycobacterium leprae is associated with defective expression of interleukin 2 receptors and is not reconstituted by interleukin 2. AB - Patients with lepromatous leprosy (LL) but not borderline tuberculoid leprosy (BT) have defective cell-mediated immune responses to Mycobacterium leprae, despite normal responses to other stimuli, as judged by in vivo skin testing and in vitro lymphocyte transformation. To investigate the basis of the immune defect in LL patients, we studied the ability of patient mononuclear leukocytes to produce interleukin 1 (IL 1) and interleukin 2 (IL 2) upon stimulation with M. leprae, and determined the ability of exogenous IL 1 and IL 2 to reconstitute the LL patient response to this antigen in vitro. Equal numbers of adherent non-T cells from LL and BT patients produced similar amounts of IL 1 upon challenge with M. leprae, and addition of IL 1 to the culture medium failed to reconstitute the response of lymphocytes from LL patients to M. leprae. On the other hand, T cells of LL patients failed to express receptors for IL 2 or to produce IL 2 in response to M. leprae, whereas similarly treated T cells of BT patients both expressed IL 2 receptors and produced IL 2. Finally, recombinant human IL 2 purified to homogeneity as well as crude supernatants of mitogen-activated lymphocytes failed to reconstitute the response of LL patients to M. leprae. These results suggest that T cells of LL patients fail to respond to M. leprae despite an ability to produce IL 1 and that their failure to express receptors for IL 2 may explain both defective proliferation and the failure of exogenous IL 2 to reconstitute the response. PMID- 2989367 TI - Polymorphic class II sequences linked to the rat major histocompatibility complex (RT1) homologous to human DR and DQ sequences. AB - Until recently, the analysis of Class II genes linked to the rat major histocompatibility complex, RT1, has been confined to serologic and electrophoretic analysis of their gene products. To obtain a more definitive estimate of the number and relative polymorphism of RT1 Class II sequences, we performed Southern blot analysis of rat genomic DNA employing human cDNA probes specific for Class II heavy and light chain genes. Southern blots of EcoRI and BamHI digests of genomic DNA from ten inbred strains, expressing eight RT1 haplotypes, were hybridized with the human DQ beta or DR beta cDNA that are homologous to Class II light chain sequences. Four to eight bands were observed to hybridize with the light chain cDNA: band sizes ranged from 2.5 to 28 kb. Restriction fragment patterns were polymorphic; the only identical patterns observed were those associated with RT1 haplotypes with identical RT1.B regions. The number and size of bands hybridizing with DQ beta and DR beta suggested a minimum of four light chain sequences in each haplotype. Southern blots of BamHI and EcoRI digests of genomic DNA from the same strains were hybridized with a DR alpha cDNA that is homologous to Class II heavy chain sequences. All RT1 haplotypes expressed either a 10.0-kb or 13.0-kb band when digested with BamHI, and either a 17-kb or 3.7-kb band when digested with EcoRI. Considerably less polymorphism was detected with the DR alpha probe; this observation is consistent with previously reported limited protein polymorphism of the rat equivalent of the I-E alpha subunit. The size and number of bands hybridizing with the DR alpha probe suggests a minimum of two heavy chain sequences. These observations suggest that the RT1 complex includes more Class II sequences than have been observed in serologic and electrophoretic analyses of Class II gene products. Furthermore, the level of polymorphism of RT1 Class II sequences appears to be comparable with mouse and human Class II sequences. PMID- 2989368 TI - Cyclic AMP induces terminal deoxynucleotidyl transferase in immature B cell leukemia lines. AB - Terminal deoxynucleotidyl transferase (Tdt), a unique DNA polymerase found only in lymphoid cells, may be involved in the generation of immunoglobulin-combining site diversity. To study the actual metabolic function of the enzyme, we developed a system in which Tdt expression is induced under defined culture conditions. We found that pharmacologic agents that raise intracellular cyclic AMP levels, such as caffeine, induce a three- to 10-fold increase in enzyme biosynthesis rate and activity. This phenomenon is observed only in pre-B cell lines of human or murine origin. PMID- 2989369 TI - Relationship between immunoglobulin production and immortalization by Epstein Barr virus. AB - After infection with Epstein Barr virus (EBV), human B lymphocytes actively secrete immunoglobulin (Ig) and are immortalized to become long-term cell lines. In these studies, we investigated the relationship between these virally induced processes utilizing limiting dilution culture techniques, and asked whether all B cells stimulated by EBV to secrete Ig are also immortalized. The activation of B cells by EBV resulting in Ig production and immortalization involved a single precursor cell, required live viral particles, and was independent of immunity to EBV by the lymphocyte donor. However, the precursor frequency of B cells activated to secrete Ig (mean 4.7%) was higher than the precursor frequency of B cells activated to long-term in vitro growth (mean 2.1%). When examined at a single cell level, it appeared that although the vast majority of the immortalized B cells also secrete Ig, only approximately 50% of the B cell precursors induced by EBV to secrete Ig go on to form long-term cell lines. In addition, although immortalized B cell clones producing all major classes of Ig were detected, IgM-committed precursors were more likely to become immortal than were precursors committed to IgG or IgA production. In contrast to these findings in B cells freshly infected with EBV, Ig production was almost always associated with evidence of long-term growth when B cells from previously established EBV induced B cell lines were tested in identical limiting dilution cultures. Thus, after infection with EBV, human B cells can either become transiently activated to proliferate and to secrete Ig, or become transformed into long-term cell lines most of which produce Ig. PMID- 2989370 TI - Lymphocyte production of endorphins and endorphin-mediated immunoregulatory activity. AB - The immune and neuroendocrine systems appear able to communicate with each other by virtue of signal molecules (hormones) and receptors common to both systems. This review summarizes our data concerning the production of one type of neuroendocrine hormone, endorphins (END), by stimulated lymphocytes. The induction by various agents and structural characterization are described, as well as the lymphocyte-derived END's potential role as a mediator of endotoxic shock. Also discussed is the ability of pituitary-derived END and the related enkephalins to inhibit a primary in vitro antibody response. These activities are discussed in the context of a complete regulatory circuit between the immune and neuroendocrine systems. PMID- 2989371 TI - Neuropeptides and their receptors: a psychosomatic network. AB - A major conceptual shift in neuroscience has been wrought by the realization that brain function is modulated by numerous chemicals in addition to classical neurotransmitters. Many of these informational substances are neuropeptides, originally studied in other contexts as hormones, "gut peptides," or growth factors. Their number presently exceeds 50 and most, if not all, alter behavior and mood states, although only endogenous analogs of psychoactive drugs like morphine, Valium, and phencyclidine have been well appreciated in this context. We now realize that their signal specificity resides in receptors (distinct classes of recognition molecules), rather than the close juxtaposition occurring at classical synapses. Rather precise brain distribution patterns for many neuropeptide receptors have been determined. A number of brain loci, many within emotion-mediating brain areas, are enriched with many types of neuropeptide receptors suggesting a convergence of information at these "nodes." Additionally, neuropeptide receptors occur on mobile cells of the immune system; monocytes can chemotax to numerous neuropeptides via processes shown by structure-activity analysis to be mediated by distinct receptors indistinguishable from those found in brain. Neuropeptides and their receptors thus join the brain, glands, and immune system in a network of communication between brain and body, probably representing the biochemical substrate of emotion. PMID- 2989372 TI - Stress, opioid peptides, the immune system, and cancer. AB - Our results indicate that a particular form of footshock stress can suppress immune function in rats and decrease their resistance to tumor challenge. These effects appear to be mediated by opioid peptides released by stress, and they can be mimicked by high doses of morphine given systemically or by a vastly smaller dose delivered intracerebroventricularly. Such findings fit well into the emerging field of behavioral neuroimmunology and reinforce continuing efforts to elucidate the neural and neurohumoral mechanisms by which the environment can affect the organism's immune system. PMID- 2989373 TI - Peptide hormones shared by the neuroendocrine and immunologic systems. AB - While numerous studies have demonstrated that the neuroendocrine system can control immune functions, it is only now becoming apparent that the control is reciprocal in that the immune system can control neuroendocrine functions. In this paper, recent studies which seem to provide a molecular basis for this bidirectional communication are reviewed. These studies suggest that the immune and neuroendocrine systems represent a totally integrated circuit by virtue of sharing a common set of hormones, such as corticotropin, thyrotropin, and endorphins, and their receptors. Possible hypothalamic and immunologic controls of this circuitry are discussed. PMID- 2989374 TI - Human and chimpanzee monoclonal antibodies. AB - Monoclonal antibody-secreting cell lines were isolated after transformation of peripheral blood leukocytes with Epstein-Barr virus. Blood samples were obtained from human donors having circulating antibodies against hepatitis viruses (HAB, HBV), rubella, or rabies virus and from a chimpanzee infected with HAV. Dextran isolated leukocytes were submitted to Epstein-Barr virus infection at low cell concentrations (1 X 10(4) cells X ml-1). Proliferating clones could be observed in 50-100% of the cultures within 4-6 weeks. Out of 1 ml blood (1 X 10(6) leukocytes) 1-10 stable clones were isolated, secreting specific anti-viral antibodies. These clones were fused with an aminopterin-sensitive, ouabain resistant, non-immunoglobulin producing mouse-human hybridoma (Org MHH.1). From such fusions 10-90% of the cultures yielded viable hybridomas of which 45% produced antibodies with the same specificity as of the parental EBV transformant. Immunoglobulin production of both EBV transformants and hybridomas was shown to be stable for more than 6 months and at a concentration up to 100 micrograms X ml-1 X 48 h-1. Chimpanzee EBV-transformed lymphocytes proliferated excellently in vitro. Mouse-human hybridomas, however, could be more easily cultivated, cloned and scaled up than the parental EBV-transformed lymphocytes. In conclusion, stable, monoclonal antibody-secreting cell lines of either human or chimpanzee origin could be isolated with an efficiency that exceeds by 10-100 fold standard murine hybridoma technology. PMID- 2989375 TI - Multifunctional inhibition by anthralin in nonstimulated and chemotactic factor stimulated human neutrophils. AB - Treatment of human polymorphonuclear leukocytes (PMN) with anthralin (0.2-50 micrograms/ml) results in dose-dependent inhibition of nondirected as well as directed migration (chemotaxis) against the synthetic tripeptide N-formyl methionyl-leucyl-phenylalanine (FMLP), the complement fragment C5a and leukotriene B4. Polymorphonuclear leukocytes (PMN) pretreated with anthralin at concentrations which inhibit cell motility also show a dose-dependent inhibition of superoxide anion generation. In contrast to anthralin two derivatives (danthrone and anthralin dimer) were ineffective. Specific binding of [3H]FMLP to neutrophil membrane receptors was impaired by anthralin at concentrations 5-10 fold higher than those which were inhibitory for cell function. Release of beta glucuronidase from azurophilic (lysosomal) granules provoked by various chemotaxins in the presence of cytochalasin B was not affected by anthralin over a wide range of concentrations. Also there were no signs of cytotoxicity e.g., leakage of cytoplasmatic lactate dehydrogenase (LDH) caused by anthralin, These data indicate that neutrophil functions may become substantially altered by anthralin. The effective dosages correspond to concentrations obtained in vivo after local application. Danthrone as well as anthralin dimer, known to be clinically ineffective, showed no effects upon PMN function. It is suggested that anthralin via a free radical mechanism alters sensitive sites at or in the cellular membrane including receptors. PMID- 2989376 TI - Immune regulation: what immunodeficiency disease has taught us. AB - "Immune regulation: what immunodeficiency disease has taught us" is reviewed by discussing three immuno-deficiency disorders. Hypogammaglobulinemia, the first documented primary immunodeficiency disorder, has a well defined and uniform clinical presentation which reflects a variety of underlying abnormalities involving the B cell, T cell, and monocyte. X-linked hypogammaglobulinemia, transient hypogammaglobulinemia of infancy common variable immunodeficiency, and their pathogenesis are discussed. Combined immunodeficiency with adenosine deaminase (ADA) deficiency first led to the now accepted concept that a biochemical abnormality may result in immunodeficiency. The clinical presentation, possible biochemical abnormalities resulting in the observed immunodeficiency, relative selectivity of the defect for the immune system, and potential applications of knowledge gained from the study of ADA deficiency are presented. Acquired immunodeficiency (AIDS) has resulted in the concept that a virus is cytopathic for a specific population of T cells and that this, at least in part, results in the immunodeficiency seen in AIDS. PMID- 2989378 TI - Molecular cloning of human and murine interleukin-2 genes and their expression in various host cells. AB - Interleukin-2 (IL-2), originally defined as a lymphokine which permits clonal expansion of T-cell clones, appears to elicit multiple biological activities in immune regulation. IL-2 cDNA cloning was one possible means to elucidate the whole structure of the IL-2 molecule and to obtain the molecule as a pure lymphokine in large quantity. The cloning and expression of human IL-2 cDNA is previously reported by us. Recently, we have obtained mouse IL-2 cDNA clone. Sequence analysis of the cloned cDNAs revealed that the murine IL-2 consists of 169 amino acids including a signal peptide. Interestingly, murine IL-2 appears to contain 12 glutamine residues in a row at the N-terminal region. Expression of this cDNA under the control of SV40 early promoter resulted in a typical mouse IL 2 activity. To reveal the structure, particularly potential regulatory elements of the IL-2 gene, we have cloned human and mouse IL-2 genes. Both genes are contained in a 7 kb genomic DNA fragment and blocked by 3 introns. The sizes of corresponding introns and exons between human and mouse are very similar to each other. The homology upstream from the TATA box is significantly higher than exon homology, implying the presence of regulatory sequence in this region. PMID- 2989377 TI - Regulation of human IgE synthesis. AB - The capacity to mount an IgE antibody response to antigen differentiates normal from allergic individuals. The frequent association of high serum IgE with T cell immunodeficiencies suggest a role for T cells in the regulation of human IgE. Peripheral blood lymphocytes (PBL) from allergic individuals spontaneously secrete IgE in vitro whereas normal PBL do not. Unlike the situation with other isotypes the polyclonal B-cell activators, pokeweed mitogen, and Epstein-Barr virus, do not induce IgE synthesis in normal B cells. However, normal B cells synthesize IgE when cultured with T cell clones that recognize determinants expressed by the B cells. B cells from allergic subjects but not from normal subjects are induced to secrete IgE via a bystander effect when cultured with T cell clones which are stimulated by antigenic determinants not expressed by the B cells. T cells with Fc receptors for IgE can be isolated from patients with hyper IgE syndrome and maintained as long term continuous T cell lines or T-T hybridomas. These cells secrete IgE binding factors which enhance IgE synthesis by preactivated IgE bearing B cells from allergic subjects but not resting B cells from normal donors. The phenotype of these Fc R+ IgE-potentiating T cell lines are predominantly T3+T4+Ia+. IgE binding factors have been isolated from normal serum and selectively suppress IgE synthesis. In contrast IgE binding factors isolated from patients with hyper IgE syndrome contain IgE-potentiating activity as well as IgE suppressor activity. These results suggest that in allergic individuals IgE B cells are activated by T cells and their antibody secretion is modulated by IgE binding factors. PMID- 2989379 TI - Human complement receptors for C3b (CR1) and C3d (CR2). AB - The human C3b receptor (CR1) is a polymorphic glycoprotein comprised of a single polypeptide chain. Of the 4 allotype forms of CR1 that have been described, the 2 most common have Mr's of 250,000 and 260,000, and are regulated by alleles having frequencies in a Caucasian population of 81.5% and 18.5%, respectively. CR1 is present on erythrocytes, neutrophils, eosinophils, monocytes, macrophages, B lymphocytes, some T lymphocytes, mast cells, and glomerular podocytes. CR1 number on erythrocytes is genetically regulated, and ranges from less than 100 sites per cell to greater than 1000 sites per cell, the average in the normal population being 500-600 sites per cell. A model accounting for this wide distribution proposes the existence of 2 codominant alleles determining low and high receptor number respectively; CR1 number is not affected by the structural polymorphism, so that the loci for these two phenotypic characteristics are distinct. The function of CR1 on erythrocytes may be to promote the clearance of immune complexes from the circulation. CR1 number on myelomonocytic cells is regulated by chemotactic factors which can rapidly transfer CR1 sites from a latent, presumably intracellular, site to the plasma membrane of these cells, thereby enhancing their ability to interact with opsonized foreign material. The receptor is involved in the endocytic reactions of these cells, and recent findings have demonstrated that this function can be modulated by T cell-derived factors, fibronectin, and phorbol esters. The role of CR1 on lymphocytes remains to be fully explored, although the receptor may enhance the differentiation of B cells into antibody-secreting cells. PMID- 2989380 TI - Viral infections and cell-mediated immunity in immunodeficient homosexual men with Kaposi's sarcoma treated with human lymphoblastoid interferon. AB - Herpesviral infections and cellular immunity were studied in 19 patients with acquired immunodeficiency syndrome who were treated with human lymphoblastoid interferon for Kaposi's sarcoma. Before treatment, cytomegalovirus (CMV) and Epstein-Barr virus were isolated from 18 of 19 patients and 13 of 14 patients, respectively. Serum levels of interferon were measurable in all cases. Concanavalin A induced lymphocyte proliferation normally in 16 of 18 patients, but CMV induced proliferation in only nine of 18 patients. Natural killer cell activity was normal in 12 of 19 patients and was augmented in vitro by interferon in six of 19 subjects. CMV-specific HLA-restricted cytotoxic T cell activity was found in only two of 15 cases. With therapy, serum levels of interferon increased in 15 of 18 patients. There were two partial tumor remissions but no improvements in viral infections. Natural killer cell activity was decreased in 11 of 14 cases, and in vitro augmentation by interferon was absent in all five previous responders. CMV-specific T cell activity did not improve, but HLA-unrestricted cytotoxicity was increased in four of eight cases. PMID- 2989381 TI - Subtypes of herpes simplex virus type 1 in Japan: classification by restriction endonucleases and analysis of distribution. AB - An attempt was made to classify herpes simplex virus type 1 (HSV-1) isolates into subtypes on the basis of the combination of the gain or loss of specific cleavage sites of HSV-1 genomes with each of three restriction endonucleases (Bam HI, Kpn I, and Sal I). According to the criteria we used for the determination of HSV-1 subtypes, 93 strains of HSV-1 that were isolated in three areas of Japan (Sapporo, Tottori, and Kagawa) were tentatively classified into eight subtypes: subtypes A-H. The bulk of the strains (84 of 93) fell into three subtypes: A, C, and H. There were highly significant differences (P less than .01) in the proportion of subtypes A and H that were isolated in Sapporo as compared with those isolated in Tottori and in Kagawa, which are geographically far from Sapporo. No significant differences, however, were found in subtypes isolated in Tottori as compared with those isolated in Kagawa, which are geographically close to each other. These data suggest that there might be a correlation between the genome structure of HSV-1 and the areas of their isolation in Japan. PMID- 2989382 TI - Phenotypic properties of atypical lymphocytes in cytomegalovirus-induced mononucleosis. AB - The phenotypic properties of the surface of the atypical lymphocytes seen in cytomegalovirus-induced (CMV) mononucleosis were evaluated. Mononuclear cells from peripheral blood were obtained from adult patients within seven to 35 days of the onset of acute CMV mononucleosis. Sheep red blood cell rosetting techniques were used to obtain populations depleted of, or enriched for, T cells. Cell populations were further purified for helper/inducer lymphocytes (T4), cytotoxic/suppressor lymphocytes (T8), or non-T-lymphocytes by monoclonal antibody binding and by complement-lysis techniques. Cytocentrifuge preparations of the cell fractions were evaluated and atypical lymphocytes were identified by morphological characteristics. The appearance and disappearance of atypical lymphocytes paralleled and antedated the increase and subsequent decrease in T8 cells seen in these patients. A total of 69% +/- 22% of the atypical lymphocytes in the lymphocyte population were of the T8 phenotype, whereas 13% +/- 10% of the atypical lymphocytes were of the T4 phenotype and 18% +/- 13% were non-T lymphocytes. Thus, atypical lymphocytes in CMV mononucleosis reside predominantly, but not exclusively, in the T8 cell population. PMID- 2989383 TI - Hepatitis A virus infection and the interferon system. PMID- 2989384 TI - Experimental Argentine hemorrhagic fever in rhesus macaques: viral strain dependent clinical response. PMID- 2989385 TI - Diagnostic significance of antibodies to the Epstein-Barr virus-specific membrane antigen gp250. PMID- 2989386 TI - The significance of specific IgA antibodies in the serum in the early diagnosis of zoster. PMID- 2989387 TI - Mechanisms of resistance of Aspergillus fumigatus Conidia to killing by neutrophils in vitro. AB - Despite the critical role for neutrophils in host defenses against invasive aspergillosis, previous studies have established that neutrophils are unable to kill resting conidia of Aspergillus fumigatus. The mechanisms of resistance of the conidia were therefore investigated. Electron microscopy studies showed the fusion of phagosomes containing A. fumigatus conidia with lysosomes of the neutrophil. Resting conidia of A. fumigatus were then compared with those that had been preincubated in broth until swollen, but not germinated, as well as with blastospores of Candida albicans (two fungal forms that are killed by neutrophils) and zymosan particles. Despite comparable susceptibility to phagocytosis, resting conidia of A. fumigatus stimulated production of significantly less superoxide anion, hydrogen peroxide, and hypochlorous acid and induced less myeloperoxidase-dependent iodination by neutrophils than did the preincubated conidia of A. fumigatus, blastospores of C. albicans, or zymosan particles. In addition, resting conidia of A. fumigatus were relatively resistant to cell-free killing by oxidants presumed to be generated by neutrophils. Thus, resistance of resting conidia of A. fumigatus to neutrophil fungicidal mechanisms appears to be secondary to both failure of the conidia to stimulate an optimal respiratory burst as well as resistance of the conidia to neutrophil oxidants. However, the reversal of this resistance by preincubation of the conidia suggests that neutrophils still may form an important host defense against the conidia of A. fumigatus. PMID- 2989388 TI - Detection of herpes simplex virus in women with acute pelvic inflammatory disease. AB - Four women are described with acute salpingitis confirmed by laparoscopy who had herpes simplex virus (HSV) isolated from the cervix or the upper genital tract (endometrium, fallopian tube, or cul-de-sac) or both. None of the patients had overt genital herpes, but one had typical HSV cervitis on a cervicovaginal smear stained with Papanicolaou's stain, one had a significant change in level of antibodies to HSV, and one had an endometrial biopsy specimen positive for HSV antigen. There are at least three potential explanations for these findings: chronic viral shedding, viral reactivation caused by acute pelvic inflammatory disease (PID), or that the PID was actually caused by HSV. Further prospective studies are needed to document the role of HSV in causing PID. PMID- 2989389 TI - In vivo antiviral activity of recombinant murine gamma interferon. AB - Recombinant murine gamma interferon (gamma-IFN) was tested for its antiviral activity in vivo. IFN preparations purified to greater than 95% purity were administered to CD-1 mice infected with lethal doses of encephalomyocarditis (EMC) virus. An initial treatment with rMuIFN-gamma administered 4 h prior to infection with virus, followed by daily treatment for 3 consecutive days significantly protected mice against EMC virus as evidenced by animal survival after 3-4 weeks post-viral infection. Variations in the antiviral effect relative to dose levels and routes of administration were also studied. PMID- 2989390 TI - Potentiation of the antiviral and anticellular activities of interferons by mixtures of HuIFN-gamma and HuIFN-alpha or HuIFN-beta. AB - Treatment of transformed human amnion WISH cells or human diploid fibroblasts (FS 4) or human fibroblasts trisomic for chromosome 21 (GM2767) with mixtures of human interferon gamma (HuIFN-gamma) and either natural leukocyte HuIFN-alpha or recombinant HuIFN alpha 2 or natural fibroblast HuIFN-beta resulted in potentiation of the antiviral activity of these IFNs. Pretreatment for 22 h of WISH cells with HuIFN-gamma followed by the addition of either HuIFN-alpha or HuIFN-beta resulted in significant potentiation of the antiviral action of these IFNs. The range of potentiation was 3 to 15-fold. Similar potentiation was observed when these IFNs were added simultaneously to the cells. Pretreatment for 22 h of WISH cells with either HuIFN-alpha or HuIFN-beta followed by the addition of HuIFN-gamma also resulted in significant increase of the antiviral protection against virus yield (3 to 39-fold). The level of the potentiation was higher in comparison with the antiviral activity observed when all these IFNs were added at the same time. Treatment of human FS-4 and GM2767 fibroblasts with HuIFN-gamma and either HuIFN-alpha or HuIFN-beta revealed potentiation of anticellular properties of these IFNs. The level of potentiation of anticellular activity was in the range of 2.7- to 9.4-fold. In the majority of the experiments, maximum potentiation of either antiviral or anti-cellular activity was observed when mixtures of equivalent concentrations of IFNs were used. The antiviral and anticellular functions of natural or recombinant IFN-alpha and fibroblast IFN beta were potentiated usually to a similar degree by the presence of IFN-gamma. In contrast, combinations of HuIFN-alpha (natural or recombinant) and HuIFN-beta, in the absence of HuIFN-gamma, did not potentiate the anticellular or antiviral activity. PMID- 2989391 TI - [Protein phosphatases]. PMID- 2989392 TI - [On the origin of spinal cord potentials evoked by peripheral nerve stimulation and its clinical meaning. Part 1: Clinical study]. AB - In the present paper, the author has attempted to show the origin of the spinal cord potential in man and to establish a practical technique for evaluating the function of the spinal cord and root. In 80 patients with spinal cord and root lesions rectangular current electrical stimulation of 0.1-0.2 msec duration and 1 3 Hz in frequency was given to the median nerve. Basic wave shape of spinal cord potential recorded from epidural space showed the two components which were the biphasic spike wave (the 1st potential) and the negative slow wave (the 2nd potential). The 1st potential elicited by stimulation of the median nerve at the elbow had a duration of 2.3 +/- 0.2 msec and negative peak latency of 7.2 +/- 0.3 msec. The 2nd potential under the same conditions had a duration of 4.1 +/- 1.3 msec and negative peak latency of 9.6 +/- 0.5 msec. The 1st potential elicited by stimulation of the median nerve at the wrist had a duration of 1.9 +/- 0.2 msec and negative peak latency of 10.4 +/- 0.7 msec. The 2nd potential under the same conditions had a duration of 6.2 msec and negative peak latency of 12.2 +/- 0.6 msec. According to the position of leading electrode, the two components changed independently in amplitude. Amplitude of the 2nd potential obtained from the midline of the epidural space was relatively larger than that from facet joints. Evoked spinal cord potentials obtained from the lateral border of facet joints revealed marked 1st potential. Recovery curves of the two components showed that the 1st potential quickly recovered to 100% in amplitude in 10 msec, but that the 2nd potential recovered slowly to only 70% in 100 msec. In cases of root lesions, spinal cord potentials revealed positivity of the 1st potential, polyphasic wave or disappearance of wave shape. In cases of spinal cord lesion, normal 1st potentials and flattening of the 2nd potentials were seen. The author has confirmed that the origin of the 1st potential in man is in primary afferent fiber and that the 2nd potentials in man reflect activity of spinal cord and are excited by synaptic relay. It is highly possible that the cervical root lesion can be differentiated from spinal cord lesion by this method. PMID- 2989393 TI - A study on the osseous elements in malignant fibrous histiocytoma of bone. AB - By studying histochemically and ultrastructurally a case of malignant fibrous histiocytoma (MFH) of bone with neoplastic osteoid elements, the distinction of the tumor from osteosarcoma was discussed. MFH was likely to originate from undifferentiated mesenchymal cells that induced a histiocytic line of differentiation, and some histiocytes seemed to be transformed into well differentiated fibroblastic cells which accordingly differentiated into fibroblasts and osteoblasts. This may explain the existence of the osseous elements in MFH. Sarcoma showing the bimodal--histiocytic and fibroblastic- differentiation should be diagnosed as malignant fibrous histiocytoma, even if it contains tumorous osteoid; it must be clearly distinguished from osteosarcoma, in which the various cells only on a fibroblastic line of differentiation can be identified and the osteoblasts are the predominant cells. PMID- 2989394 TI - [Tissue reactions to calcium phosphate ceramics used as root implants in the alveolar bone of dogs]. PMID- 2989395 TI - Renal and hormonal responses to acute hypoxia in normal individuals. AB - We studied, in normal volunteers, the effects of 1 hour of hypoxia on the concentration of angiotensin-converting enzyme and bradykinins, along with previously measured parameters of renal and endocrine function. Ten men, 18 to 42 years of age, undergoing water diuresis, breathed a low-oxygen mixture (five breathed 10.5% O2 and five 12% O2); all breathed 21% O2 on a control day. Measurements included mean blood pressure and heart rate every 2 to 3 minutes; plasma levels of renin activity, aldosterone, arginine vasopressin, norepinephrine, and bradykinin, and angiotensin-converting enzyme activity, before and at the end of gas breathing; and urine volume (UV), creatinine, Na+, and bradykinin concentration. Arterial blood gases and effective renal plasma flow were determined at the end of gas breathing only. Mean values +/- SEM for arterial blood gases with low O2 were pH 7.39 +/- 0.02, PO2 46 +/- 2 torr, PCO2 39 +/- 2 torr (12% O2) and 7.48 +/- 0.01, 35 +/- 1 torr, 33 +/- 1 torr (10.5% O2). Responses were otherwise identical in both groups, and data were combined for analysis. With hypoxia, heart rate and effective renal plasma flow increased significantly, P less than 0.005; no changes occurred in Uv, urine Na+ concentration, glomerular filtration rate, plasma or urine bradykinin concentration, serum angiotensin-converting enzyme activity, plasma renin activity, plasma aldosterone concentration, plasma arginine vasopressin concentration, or plasma norepinephrine concentration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989396 TI - Sodium-potassium-adenosine triphosphatase in nephron segments of spontaneously hypertensive rats. AB - Sodium pump activity of blood vessels has been reported to decrease in several animal models of hypertension. We studied sodium-potassium-adenosine triphosphatase (Na-K-ATPase) activity of renal tubular segments in 12-week-old spontaneously hypertensive rats and in age-matched Wistar-Kyoto normotensive rats. The enzyme activity of the individual nephron segments was determined by a microfluorometric assay in which ATP hydrolysis is coupled with NADH oxidation. In the spontaneously hypertensive rats, systolic blood pressure was significantly higher (181 +/- 3 mm Hg) than in the Wistar-Kyoto rats (134 +/- 2 mm Hg). However, there was no difference in mean Na-K-ATPase activity in any of the nephron segments from the spontaneously hypertensive compared with the Wistar Kyoto group. It is concluded that Na-K-ATPase activity does not change in any of the nephron segments with spontaneous hypertension. PMID- 2989397 TI - Demonstration of functional low-density lipoprotein receptors by protein blotting in fibroblasts from a subject with homozygous receptor-negative familial hypercholesterolemia. AB - We report the detection of low-density lipoprotein (LDL) receptors by the technique of receptor blotting in fibroblasts from a patient with homozygous familial hypercholesterolemia (FHC) previously classified as "receptor negative." Solubilized receptors were electrophoresed, transferred to nitrocellulose paper, treated with LDL followed by radiolabeled antibody to LDL, and visualized by autoradiography. GM 2000 FHC fibroblasts revealed LDL receptors with an apparent molecular weight of approximately 140,000, the same as in normal cells. LDL receptor activity by blotting in GM 2000 cells was greatly diminished in comparison with normal cells, but was calcium dependent. Receptor activity was also detectable by conventional monolayer binding and degradation assays. Thus, GM 2000 cells have profoundly diminished LDL receptor activity, but retain the genetic capacity to make LDL receptor material of normal molecular weight that is capable of binding LDL. Previous studies have demonstrated the presence of trace amounts of immunoreactive LDL receptor protein in fibroblasts from some receptor negative FHC homozygotes. Our work extends these studies by demonstrating the ability of this material to bind LDL. PMID- 2989398 TI - Long-term follow-up of IgG and IgA antibodies against viral capsid antigens of Epstein-Barr virus in nasopharyngeal carcinoma. AB - A total of 137 patients with biopsy-proved anaplastic epidermoid nasopharyngeal carcinoma (NPC) seen at the National Taiwan University Hospital from December 1971 through December 1973, were studied serologically before radiotherapy and during the follow-up period up to December 1982. A clear control study on 134 healthy patients or patients with diseases other than NPC was also done. EB virus associated anti-VCA antibodies in both IgG and IgA classes were titrated by means of indirect immunofluorescent antibody method on two to six samples of serum from the patients during the follow-up study. When seropositive standard was set at 1:640 for anti-VCA/IgG and 1:40 for anti-VCA/IgA, the seropositive rates were 63.5 per cent and 81.1 per cent for NPC patients before treatment and 2.9 per cent and 2.2 per cent for the control respectively. The differences of seropositive rates between the patients and control were statistically highly significant, as chi 2 greater than 111, p less than 0.0005. At the completion of radiotherapy and during the following year, some reduction and fluctuation of seropositive rates were seen in both cured and recurrent patients. From the second year after radiotherapy and thereafter, the seropositive rates were 72 per cent-100 per cent for patients with recurrences and 17.6 per cent-30.8 per cent for cured patients. The differences were significant (chi 2 greater than 24.8, p less than 0.0005). Therefore, high titres of anti-VCA antibodies may coexist with cancer tissue in NPC patients. IgA class antibodies is slightly higher in sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989399 TI - Pleomorphic adenoma of the larynx. AB - A case of pleomorphic adenoma of the subglottic larynx in a 69-year-old female with unusual presentation is reported. The origin, histopathology and treatment of pleomorphic adenomas are discussed. PMID- 2989400 TI - Isolation and characterization of cDNA clones corresponding to two different human apoC-III alleles. AB - We have recently reported that the human apolipoprotein A-I (apoA-I) and apolipoprotein C-III (apoC-III) genes are physically linked and that the presence of a DNA insertion in the apoA-I gene is correlated with apoA-I-apoC-III deficiency in patients with premature atherosclerosis. In addition, the presence of a polymorphic restriction endonuclease site (SacI) in the 3' noncoding region of apoC-III mRNA has been correlated with hypertriglyceridemia in humans. In this study, we report the isolation and characterization of cDNA clones containing the entire apoC-III mRNA coding sequence. The nucleotide-derived apoC-III amino acid sequence indicates that the apoC-III primary translational product contains a 20 amino acid N-terminal extension, which conforms with the general properties of known signal peptides, and is highly homologous to the recently reported rat apoC III signal peptide. The DNA-derived apoC-III amino acid sequence differs from the previously reported apoC-III amino acid sequence at four amino acid residues. More specifically, at positions +32, +33, +37, +39, the DNA sequence predicts Glu, Ser, Gln, Ala, respectively, while the previously reported sequence specifies Ser, Gln, Ala, Gln, respectively. Finally, isolation and characterization of apoC-III cDNA clones, with or without the polymorphic SacI restriction site, indicated that the apoC-III nucleotide sequence corresponding to the Sac+ and Sac- clones differs at three nucleotide sites; however, the amino acid sequence specified by the Sac+ and Sac- alleles is identical. PMID- 2989401 TI - A comparative study of eicosapentaenoic acid metabolism by human platelets in vivo and in vitro. AB - During long-term dietary n-3 fatty acid supplementation, eicosapentaenoic acid (EPA) is not incorporated into phosphatidylinositol or -serine of human platelets in vivo and is not detectable in phosphatidic acid upon stimulation with thrombin. However, EPA is released from platelet phospholipids and metabolized to thromboxane B3 (TXB3). In contrast, in vitro, platelets incorporate [14C]EPA into phosphatidylinositol, whether they contain endogenous EPA in their cellular lipids or not. Following platelet stimulation, [14C]EPA appears in phosphatidic acid, as free fatty acid, and is transformed to TXB3. We conclude that the fatty acid compositions of platelet phospholipid subclasses are regulated with a high degree of specificity in vivo. Qualitative differences exist between in vivo and in vitro uptake of EPA into platelet phospholipid subclasses. After in vivo incorporation, EPA is released by action of a phospholipase A2. PMID- 2989402 TI - Alkaline phosphatase activity can interfere with the enzymatic determination of triglycerides. AB - Unlike human plasma, rat plasma was found to contain substantial quantities of alkaline phosphatase. The large amount of phosphatase activity in rat plasma interferes with assay methods that employ orthophosphoric acid esters as substrates or reaction intermediates. Although these methods are effective when applied to human plasma samples, they cannot be used with rodent plasma. PMID- 2989403 TI - Trophoblastic tumors. PMID- 2989404 TI - Structure of the penicillin acylase gene from Escherichia coli: a periplasmic enzyme that undergoes multiple proteolytic processing. AB - Penicillin acylase is processed from a 90-kD precursor through the cleavage of a leader peptide and two further endopeptidase cleavages to yield an enzyme that contains a 22-kD (or 23-kD) and a 65-kD subunit. The endopeptidase cleavages require an intact carboxy terminus. This type of processing appears to be unique for a prokaryotic enzyme, having its most closely related analog in the synthesis and processing of preproinsulin and other eukaryotic hormones. PMID- 2989405 TI - A chlorophyll a/b-protein encoded by a gene containing an intron with characteristics of a transposable element. AB - We have sequenced a genomic subclone (pLg AB19/H5c) of Lemna gibba nuclear DNA containing a complete chlorophyll a/b protein coding region and 5' and 3' flanking nucleotides. The coding region contains an intron of 84 nucleotides that has features characteristic of a transposable element. Evidence from S1 nuclease mapping experiments is consistent with correct transcription and splicing of the AB19 or another closely related intron-containing gene. The encoded precursor polypeptide of 264 amino acid residues has a predicted Mr of 28,327. Approximately 35 N-terminal residues are cleaved from this protein to form the mature apoprotein. We have used theoretical considerations of protein structure to propose an experimentally testable model of the structure of this protein in thylakoid membranes. PMID- 2989406 TI - Comparison of the transient late region expression of SV40 DNA and SV40-based shuttle vectors: development of a new shuttle vector that is efficiently expressed. AB - Transient expression of an SV40-based shuttle vector similar in design to commonly used vectors is shown to be inefficient when compared to expression of SV40 viral DNA. To eliminate this problem, we have designed and constructed a new vector, pSVPiC, which contains a minimal noninterfering plasmid component, the 885-bp plasmid PiAN7, and two SV40 promoter/origin regions. Transient expression of the SV40 late region from pSVPiC is much more efficient than that from previously used vectors and even more efficient than that from SV40 viral DNA. When the gene for rabbit beta-globin is placed in the late region of pSVPiC, it is also expressed at high levels, indicating that this shuttle vector is generally useful for expressing eukaryotic genes. PMID- 2989407 TI - A quantitative description of flagellar movement in golden hamster spermatozoa. AB - Flagellar movement of golden hamster spermatozoa obtained from the testis and the caput and cauda epididymides was observed by a light microscope while holding them at their heads with a micropipette. Flagellar movement of capacitated spermatozoa and of reactivated spermatozoa demembranated with Triton X-100 was also observed. Testicular and caput epididymal spermatozoa showed weak movement in Tyrode's solution, whereas cauda epididymal spermatozoa showed vigorous movement. The flagellar bends of the cauda epididymal spermatozoa were almost planar. Capacitated spermatozoa moved with waves of a large amplitude. Demembranated spermatozoa reactivated with ATP only had a latent period before the initiation of flagellar movement, and beat at low frequency, whereas demembranated spermatozoa reactivated with both ATP and cAMP began to move immediately at high frequency. Thrust and hydrodynamic power output were calculated using the parameters for the typical waveforms of cauda epididymal spermatozoa before and after capacitation. The possible role of the large amplitude beat in capacitated spermatozoa is discussed. A comparison of the 'principal' and 'reverse' bends in golden hamster sperm flagella as defined by Woolley (1977) with those in sea urchin sperm flagella suggests that the so called 'principal' bend in golden hamster sperm flagella corresponds to the reverse bend in sea urchin sperm flagella and vice versa. PMID- 2989408 TI - Sequential expression of genes involved in human T lymphocyte growth and differentiation. AB - Nuclear transcription assays were performed with isolated nuclei from human peripheral blood T lymphocytes stimulated with phytohemagglutinin and phorbol myristate acetate to determine the kinetics of transcriptional activity of various genes occurring in T cell activation. Although silent in resting T cells, the genes encoding c-myc and the interleukin 2 (IL-2) receptor were induced early, preceding gamma interferon (IFN-gamma), IL-2, and transferrin receptor gene transcription. Transcriptional activity of these genes fell after their respective peaks, indicating that the expression of these genes is a transient event during T cell activation. With the exception of the transferrin receptor gene, the kinetics of induction of these genes were not altered by concentrations of cycloheximide that inhibited protein synthesis. These data indicate that the induction of genes encoding c-myc, IL-2, IL-2 receptor, and IFN-gamma occur independently of the sequential production of the proteins they encode. PMID- 2989409 TI - Pertussis toxin inhibition of chemotactic factor-induced calcium mobilization and function in human polymorphonuclear leukocytes. AB - Chemotactic factors stimulate a rapid increase in the cytosolic concentration of intracellular calcium ions ([Ca2+]in) in human polymorphonuclear leukocytes (PMNL), which may be an event that is critical to the expression of chemotaxis and other PMNL functions. Treatment of PMNL with pertussis toxin catalyzes ADP ribosylation of a protein similar or identical to the inhibiting regulatory protein of adenylate cyclase, Gi, and suppresses the increase in [Ca2+]in elicited by leukotriene B4(LTB4) and formyl-methionyl-leucyl-phenylalanine. Chemotactic migration and lysosomal enzyme release elicited by chemotactic factors were inhibited by pertussis toxin with a concentration-dependence similar to that for inhibition of the increase in [Ca2+]in, without an effect on lysosomal enzyme release induced by the ionophore A23187 and phorbol myristate acetate. Activated pertussis toxin catalyzed the [32P]ADP-ribosylation of a 41 kD protein in homogenates of PMNL. The extent of [32P]ADP-ribosylation of this protein was reduced 59% by pretreatment of intact PMNL with pertussis toxin. Pertussis toxin selectively decreased the number of high-affinity receptors for LTB4 on PMNL by 60% without altering the number or binding properties of the low affinity subset of receptors. Pertussis toxin modification of a membrane protein of PMNL analogous to Gi thus simultaneously alters chemotactic receptors and attenuates the changes in cytosolic calcium concentration and PMNL function caused by chemotactic factors. PMID- 2989410 TI - Ig heavy chain variable region gene complex of lupus mice exhibits normal restriction fragment length polymorphism. AB - B cell hyperactivity, hypergammaglobulinemia, and autoantibody expression, the hallmarks of systemic lupus erythematosus, might be associated with structural abnormalities within the Ig heavy chain variable region (Igh-V) gene complex. The Igh-V loci from several lupus-prone mouse strains, their ancestors, and other nonautoimmune mice were therefore analyzed by restriction fragment length polymorphisms with DNA probes corresponding to seven VH gene families. These seven families comprise the majority of the known polymorphic murine VH gene repertoire, including some involved in autoantibody generation. Our study showed that the Igh-V loci from lupus and haplotype-matched nonlupus mice resulted in essentially identical restriction fragment patterns, a finding which suggests that the Igh-V gene complex does not carry a primary defect responsible for autoimmune disease. PMID- 2989411 TI - Stable expression of cDNA encoding the human interleukin 2 receptor in eukaryotic cells. AB - Human interleukin 2 (IL-2) receptor cDNA derived from HUT 102B2 cells was stably expressed in murine L cells. These L cell transfectants (a) displayed surface receptors of the aberrant size of the IL-2 receptors on HUT 102B2 cells, (b) did not respond to exogenous IL-2 with augmented proliferation, and (c) expressed low affinity but not high affinity receptors for IL-2. PMID- 2989412 TI - Human T cell leukemia/lymphoma virus I infection and subsequent cloning of normal human B cells. Direct responsiveness of cloned cells to recombinant interleukin 2 by differentiation in the absence of enhanced proliferation. AB - A human T cell leukemia/lymphoma virus (HTLV)-I-infected B cell clone expressed Tac antigen on its cell surface and responded to recombinant interleukin 2 (IL-2) by increased production of IgM without any increase in proliferation. Anti-Tac antibody completely inhibited the IL-2-induced differentiation of this HTLV-I infected B cell clone. This study demonstrates that HTLV-I can directly infect normal mature human B cells, and that the Tac antigen, which may be induced by infection with HTLV-I, is the functional receptor for IL-2-induced B cell differentiation. The availability of such cell lines and clones should provide useful tools to delineate precisely the differentiation step in the human B cell cycle. PMID- 2989414 TI - Spontaneous configuring in conditioned flavor aversion. AB - A series of experiments investigated spontaneous configuring using the conditioned flavor aversion paradigm with rat subjects. In Experiment 1, extended training of a two-flavor compound stimulus did not produce spontaneous differentiation of conditioned responding to that compound and its elements. In Experiment 2 we found that extended nonreinforced exposure to a compound stimulus generated spontaneous element-compound differentiation when the elements were later conditioned. Rats that received extended preexposure to the compound showed less conditioned responding to the compound than to either of its elements. However, rats that had not received preexposure to the compound showed greater conditioned responding to the compound than to either of its elements (summation). In Experiment 3, nonreinforced preexposure to the compound stimulus prior to minimal reinforced compound training produced spontaneous compound element differentiation, but extended reinforced compound training eliminated that differentiation. In Experiment 4, extended partial reinforcement training with a compound produced differentiation of the compound from its elements. Implications of these data for the mechanisms responsible for spontaneous configuring and for the summation assumptions common to most learning theories are discussed. PMID- 2989413 TI - Early events in Epstein-Barr virus infection provide a model for B cell activation. AB - We have used Epstein-Barr virus (EBV) infection in vitro to delineate two distinct stages in B cell activation. Previous studies have shown that the BLAST 2 (EBVCS) (EBV cell surface) activation antigen is expressed on a small fraction of B cells within 24 h of stimulation with a variety of agents, including mitogens and EBV. In this study, we have been able to isolate the BLAST-2 (EBVCS)+ cells early after activation/infection with EBV. These cells are small B cells that are actively synthesizing RNA but not DNA, and are, therefore, clearly distinct from large proliferating lymphoblasts. In addition, they contain multiple copies of the EBV genome, express the viral nuclear antigen (EBNA) and, most importantly, proceed to undergo transformation when placed back in culture. By comparison, the BLAST-2 (EBVCS)- population does not undergo transformation, even though a fraction of these cells are activated for RNA synthesis and express EBNA. Thus, using the EBV system, we have been able to show directly that an activated B cell first expresses the BLAST-2 (EBVCS) antigen concomitant with an increase in RNA synthesis, and then subsequently proceeds to differentiate into a proliferating lymphoblast. PMID- 2989415 TI - Quantitative analysis of lymphocyte populations and T cell subsets in patients with intracranial glioma. PMID- 2989416 TI - Two electropherotypes of bluetongue virus serotype 2 from naturally infected calves. AB - The first isolation of bluetongue virus (BTV) serotype 2 in the U.S.A. was in 1982 from a sentinel herd of cattle at Ona, Florida. Electrophoretic analysis of genome RNA revealed that all 16 serotype 2 isolates obtained from this focus of infection had one of two electropherotypes (designated Ona A and Ona B). All genome segments of Ona A and Ona B had detectable differences in electrophoretic mobility, with major differences noted for segments 1, 4, 5, 7, 8, 9 and 10. Electrophoretic comparison revealed that Ona A was indistinguishable from the African serotype 2 prototype isolated 23 years earlier. In 1983, Ona B, in the apparent absence of Ona A, was isolated from two additional cattle herds in Florida. These results suggest that Ona B may be a variant of Ona A that evolved as a result of mutation or reassortment with another BTV strain, and may be better adapted to the selective pressures found in southern Florida. Comparison of the electropherotypes of Ona B with two Florida isolates of serotype 13 and 17 and the prototypes of BTV 10, 11, 13 or 17 produced no evidence for reassortment between Ona A and any of these strains as the possible origin of Ona B. PMID- 2989417 TI - Isolation and characterization of deletion mutants of herpes simplex virus type 2 (strain HG52). AB - We provide evidence that: (i) two variants lacking the XbaI site at map coordinate 0.7 have been selected following XbaI treatment of the DNA of herpes simplex virus type 2 strain HG52; (ii) one of these mutants had lost the 0.7 restriction site due to a deletion of approximately 150 base pairs and in the other the site loss was due to a similar sized sequence insertion; (iii) following XbaI treatment, four variants with deletions ranging in size from 1.5 kb (in both TRL and IRL) to 9 kb in IRL were isolated; (iv) substantial deletions in the long terminal repeat regions of HG52 are present with a frequency of 24% of genomes in the elite stock, a variant with a 3.75 kb deletion in IRL making up 10% and one with a 1.5 kb deletion in both IRL and TRL making up 14%; (v) two of the variants isolated after XbaI treatment of viral DNA were identical to the deletion prototype within the elite stock, suggesting that these variants were not generated as a result of XbaI treatment but pre-existed in the viral DNA pool; (vi) the deletion variants were stably maintained during routine stock propagation, were viable and could be propagated as cloned populations; (vii) the deletions did not have a marked deleterious effect on the one-step growth kinetics of the virus. PMID- 2989418 TI - Detection of enteric adenoviruses by dot-blot hybridization using a molecularly cloned viral DNA probe. AB - Enteric adenoviruses (EAds) (candidate adenoviruses 40 and 41, subgroups F and G) have been implicated in the etiology of gastroenteritis in infants, but their clinical significance has been unclear because a rapid test to distinguish these agents from other adenovirus (Ad) types has not been available. We developed a dot-blot hybridization assay for EAd DNA using a cloned DNA fragment that has little homology to non-EAd DNAs. The dot-blot system detected less than 20 pg of EAd DNA, while showing minimal cross hybridization to representative strains from all other Ad groups. There was no detectable hybridization to extracts of samples known to contain other enteric viruses. It was further shown that low levels of EAds in specimens could be amplified by culturing for 1 day in 293 cells. Stool samples and tissue culture lysates prescreened by electron microscopy, cell culture or ELISA were tested in a blind fashion. Using endonuclease analysis as the standard for typing the isolates, we found the dot-blot system to have a 91% sensitivity and 71% specificity for detecting EAds and distinguishing them from other Ads. False-positive and equivocal dot-blot results appeared to be caused by other Ads. PMID- 2989419 TI - Clinical efficacy of a herpes simplex subunit vaccine. AB - A DNA-free herpes simplex type 2 subunit vaccine was administered to 18 volunteers without past evidence of herpes simplex type 1 (HSV 1) or herpes simplex type 2 (HSV 2) infection, to 44 patients with severe recurrent genital HSV 2 infection, and to 15 patients with severe oral type 1 HSV recurrences. The vaccine elicited both humoral and cell-mediated immunity in 97% of the subjects without past HSV infections and boosted significantly the cell-mediated immunity and antibody titers in almost all the patients with recurrent HSV 1 or HSV 2. The vaccine elicited particularly the production of complement-dependent cytotoxic antibodies in 96% of the patients with recurrent HSV 2 infections. This might, at least partly, explain the clinical efficacy of the vaccine. Indeed, we observed a significant decrease (t test, p less than 0.01) in the attack rate of the recurrences and also a significant shortening of the time needed to complete healing of the lesions (t test, p less than 0.01). PMID- 2989420 TI - Rotavirus prevalence and relationships with climatological factors in Gabon, Africa. AB - A one-year study of rotavirus prevalence was carried out using electron microscopy in Gabon, equatorial Africa. One hundred fifty-six nonhospitalized diarrheic children up to 10 years of age and 115 age-matched controls were investigated together with diarrheic adults and children over 10 years of age. Rotaviruses were observed only in diarrheic children under 10 years of age (10.2%). Rotaviruses were more prevalent in the dry-season months (19.2% versus 1.3%, P less than .001). Rotaviruses were the most commonly found among all the viral, bacterial, and parasitic agents we detected, with the exception of the coronaviruslike particles, which are the subject of another paper [Sitbon, 1985]. PMID- 2989422 TI - Intrathecal synthesis of IgG antibodies to varicella-zoster virus in two cases of acute aseptic meningitis syndrome with no cutaneous lesions. AB - IgG antibodies to varicella-zoster virus (VZV) were detected by indirect enzyme immunoassay (EIA) in CSF of two patients with acute aseptic meningitis syndrome (AAMS) not associated with evident cutaneous lesion or recent history of zoster infection. Their characteristic features and serological data are compared with those observed in two patients with AAMS and zoster cutaneous lesions, and in 13 patients with AAMS of unknown or other etiology. According to several indexes applied to assess the origin of the detected antibodies, it is concluded that VZV IgG antibodies are of intrathecal production. The possible etiologic relationship between the neurological syndrome and the detection of VZV antibodies in CSF is discussed. Routine serological procedures are proposed for the diagnosis of CNS acute infections of probable viral etiology that should provide additional data on these rarely described cases. PMID- 2989421 TI - Diagnosis of parainfluenza virus infection in children and older patients by detection of specific IgM antibody. AB - The significance of detecting specific antibody of the IgM class for the diagnosis of parainfluenza infections was examined. Paired sera from 763 children and adults admitted to the hospital for acute respiratory disease were tested for significant antibody titer rises in the hemagglutination inhibition (HI) test and for specific IgM antibody with the hemadsorption immunosorbent techniques (HIT). Sera were collected during two 6-month periods, January through June, 1982 and 1983. Evidence of parainfluenza infections was found in 122 patients (16%): 83 (25%) in 1982 and 39 (9.1%) in 1983. The HIT was superior to the HI test for detection of parainfluenza infection, in particular in infants and aged patients, 94 patients were positive only in the HIT test, 12 in the HI test, and 16 in both tests. In a control group of 120 persons (time- and age-matched to the patients of 1982) admitted for nonrespiratory illness, six (5%) showed parainfluenza IgM in their serum. Blocking experiments and retrospective clinical information indicated that the IgM antibody detected in these individuals is specific IgM acquired after a mild parainfluenza infection. Most (66%) patients showed IgM antibody titer rises or high titers (greater than 1,280) in both sera, and in 23%, a fall in IgM antibody titer was found. Detection of specific IgM antibody by HIT permitted early presumptive diagnosis in 71% of the patients with parainfluenza infection. IgM antibody persisted for 2-11 weeks. The HIT appears to be an important supplement for the diagnosis of parainfluenza infections. PMID- 2989423 TI - Antibody response to Epstein-Barr-virus-specific DNase in 13 patients with nasopharyngeal carcinoma in Taiwan: a retrospective study. AB - Serum samples obtained from 13 individuals who were found to have a previous history of or to be suffering from nasopharyngeal carcinoma (NPC) were examined for the presence of antibody to Epstein-Barr virus (EBV) DNase activity. Significant to high levels of antibody to EBV DNase activity were detected in most serum samples obtained from four patients prior to the diagnosis of NPC. The samples from the other nine patients showed variable levels of antibody. The majority of the samples collected at the remission stage of the disease, especially those from long-term survivors, contained little or no antibody to the DNase activity. Data presented here suggest that antibody to EBV DNase activity may be a useful marker for the early diagnosis as well as the prognosis of NPC. PMID- 2989424 TI - Behavioral recovery associated with central nervous system regeneration in the snail Melampus. AB - The pulmonate snail Melampus bidentatus regenerates central nervous tracts following commissurotomy, connective transection, and cerebral ganglion ablation. Our goal was to determine whether or not neural regrowth within the central nervous system restored behaviors disrupted by lesions. One behavior that is disrupted by commissurotomy is retraction of facial structures that are contralateral to a stimulated facial region, a response that normally accompanies the ipsilateral retraction. Tentacle withdrawal on the side contralateral to stimulation reappeared on a timescale that was correlated with growth of a commissural link (8-19 days post-lesion). Electrophysiological recordings from a labial nerve pathway that has a contralateral component similar to the contralateral tentacle response showed that development or strengthening of an alternative pathway could also mediate contralateral responses. Thus, a major conclusion of this study was that both tract regeneration and changes in existing CNS pathways can underlie recovery. The percentage (approx. 75%) of snails that regenerate the cerebral commissure and show behavioral recovery is established early in the period following commissure transection. Behavioral recovery and anatomical evidence of regeneration were also correlated in the other two operations: single cerebral ganglion removal and unilateral cerebropleural and cerebropedal connective transection. We conclude that Melampus is able to regenerate neuronal connectivity that can restore normal behavior. PMID- 2989425 TI - Octopamine enhances neuromuscular transmission in developing and adult moths, Manduca sexta. AB - The effect of octopamine on neuromuscular transmission was examined in developing and adult Manduca sexta. Intracellular recordings were made from the dorsal longitudinal muscle (DLM), superfused with solutions containing DL-octopamine or other amines. In untreated adult moths and pharate adults nearly ready to enclose (stage Day 19), stimulation of the motor nerve evokes a large excitatory junction potential (EJP), an active membrane response, and a twitch. In adults and Day 19 animals DL-octopamine (10(-7) to 10(-4)M) has no effect on the amplitude and rise time of the electrical response in normal saline, but 10(-6) to 10(-4) M DL octopamine increases the amplitude of the excitatory junction potential recorded in saline containing one-third the normal calcium concentration. Immature (Day 16) muscle, which normally produces only small EJPs following stimulation of its motor nerve, responds to 10(-6) to 10(-4) M DL-octopamine by an increase in the EJP above threshold for an active membrane response and a contraction. When the muscle has developed sufficiently to spike and contract in response to nerve stimulation in the absence of exogenous octopamine (Days 17 and 18), application of DL-octopamine increases the maximum rate at which the muscle contracts in response to each stimulus in a train (designated the maximum following frequency, MFF). The threshold dose for an effect on the MFF of Day 18 immature moths is less than 10(-10) M. At this stage 10(-8) M DL-octopamine increases the MFF four fold. The effect on the MFF is dose-dependent over the range 10(-10) M to 10(-6) M. The biogenic amines DL-epinephrine, DL-norepinephrine, tyramine, DL phenylethanolamine, 2-phenylethylamine, and dopamine, applied at concentrations of 10(-8) or 10(-4) M, do not change the MFF. Both DL-synephrine (10(-8) M) and serotonin (10(-7) M) mimic the action of 10(-10) M DL-octopamine on the MFF. The action of DL-octopamine (10(-7) M) is blocked by phentolamine (10(-4)M) but not by propranolol (10(-4)M). The octopamine content of hemolymph was determined with a radioenzymtic assay. The concentration of octopamine in the hemolymph increases 3.6-fold, from 5 X 10(-8) M on Day 18 (duration of adult development is 19 days) to 1.85 X 10(-7) M one day following eclosion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2989426 TI - Kinetics of dithionite reduction of the heme nonapeptide of cytochrome c. AB - The kinetics of dithionite reduction of the oxidized heme nonapeptide fragment of horse heart cytochrome c have been measured as a function of ionic strength at pH 7 and pH 9 by the stopped-flow technique. Dithionite concentration dependences indicate that the radical anion monomer, SO2-., is the active reductant. The pH 7 ionic strength dependence suggests that the heme peptide is reacting as a negatively charged molecule (its overall charge is calculated to be -1). Comparison of these results with the known rate of dithionite reduction of cytochrome c indicates that the heme nonapeptide has substantially greater inherent reactivity than cytochrome c, perhaps due to the greater accessibility of the heme. PMID- 2989427 TI - Various types of thyrotropin-releasing hormone receptors in discrete brain regions and the pituitary of the rat. AB - Receptors for thyrotropin-releasing hormone (TRH) in the rat brain and the pituitary are heterogenous. The receptors were classified into four types according to the dissociation constant (KD). High-affinity receptors (KD less than 3 nM) are present in the pituitary, hypothalamus, amygdala, and limbic forebrain which contains the nucleus accumbens and the septum. Intermediate affinity receptors (KD, 5-16 nM) are evidently present in the frontal cortex, hippocampus, striatum, thalamus, and the brainstem, but may also be present in other regions. Low-affinity TRH receptors (KD, 50-80 nM) are seen in the limbic forebrain, amygdala, and the hypothalamus. Very-low-affinity receptors (KD, 215 nM) exist in the pituitary. Experiments using DN-1417 (gamma-butyrolactone-gamma carbonyl-histidyl-prolinamide citrate), a synthetic TRH analogue with a more potent central activity, indicated the presence of TRH receptors having a high affinity to DN-1417 at least in the limbic forebrain but not in the pituitary. This type of receptor is not labeled by [3H](3-methyl-histidine2)-TRH. Density of the TRH receptor is the highest in the pituitary and next highest in the amygdala. PMID- 2989429 TI - Decreased incorporation of [3H]inositol and [3H]glycerol into glycerolipids of sciatic nerve from the streptozotocin diabetic rat. AB - The incorporation of [3H]myo-inositol into individual phosphoinositides and of [3H]glycerol into glycerolipids was determined in sciatic nerve obtained from normal and streptozotocin diabetic rats and incubated in vitro. The uptake of inositol into lipid was approximately linear with time. More than 80% of the label was present in phosphatidylinositol with the remainder divided about equally between phosphatidylinositol phosphate and phosphatidylinositol-4,5 bisphosphate. Labeling was unchanged 2 weeks after induction of diabetes, but was reduced by 32% after 20 weeks of the disease. Glycerol incorporation occurred primarily into phosphatidylcholine and triacylglycerol and was depressed up to 45% into major phosphoglycerides in nerves from both 2- and 20-week diabetic animals. Triacylglycerol labeling was also substantially decreased, and the reduction was comparable in intact and epineurium free nerve, suggesting that a metabolically active pool of this compound, which is sensitive to hyperglycemia and/or insulin deficiency, is located in or immediately adjacent to the nerve fibers. The considerable decline in incorporation of these lipid precursors in diabetic nerve may be related to impaired inositol transport and to decrease overall energy utilization by the tissue. PMID- 2989430 TI - Solubilization of an adenosine uptake site in brain. AB - Procedures are described for the solubilization of adenosine uptake sites in guinea pig and rat brain tissue. Using [3H]nitrobenzylthioinosine [( 3H]NBI) the solubilized site is characterized both kinetically and pharmacologically. The binding is dependent on protein concentration and is saturable, reversible, specific, and high affinity in nature. The KD and Bmax of guinea pig extracts are 0.13 +/- 0.02 nM and 133 +/- 18 fmol/mg protein, respectively, with linear Scatchard plots obtained routinely. Similar kinetic parameters are observed in rat brain. Adenosine uptake inhibitors are the most potent inhibitors of [3H]NBI binding with the following order of potency, dilazep greater than hexobendine greater than dipyridamole. Adenosine receptor ligands are much less potent inhibitors of binding, and caffeine is without effect. The solubilized adenosine uptake site is, therefore, shown to have virtually identical properties to the native membrane site. The binding of the adenosine A1 receptor agonist [3H]cyclohexyladenosine [( 3H]CHA) to the solubilized brain extract was also studied and compared with that of [3H]NBI. In contrast to the [3H]NBI binding site [3H]CHA binds to two apparent populations of adenosine receptor, a high affinity site with a KD of 0.32 +/- 0.06 nM and a Bmax of 105 +/- 30 fmol/mg protein and a lower-affinity site with a KD of 5.50 +/- 0.52 nM and Bmax of 300 +/- 55 fmol/mg protein. The pharmacology of the [3H]CHA binding site is consistent with that of the adenosine receptor and quite distinct from that of the uptake [( 3H]NBI binding) site. Therefore, we show that the adenosine uptake site can be solubilized and that it retains both its binding and pharmacologic properties in the solubilized state. PMID- 2989428 TI - Effects of 5,5'-dithiobis-(2-nitrobenzoic acid) on opiate binding to both the membrane-bound receptor and the partially purified opiate receptor. AB - Pretreatment of partially purified opiate receptor from rat brains with 5,5' dithiobis-(2-nitrobenzoic acid) (DTNB) decreased opiate agonist binding more effectively than that of antagonist. This agent, at a concentration that inhibits only 3H-agonist binding, increases the IC50 values of agonists but not those of antagonists. We also observed similar effects of DTNB on opiate binding to the membrane-bound receptor that are in good agreement with the published data. Moreover, there was an excellent correlation between the IC50 values of the two different preparations. However, opiate binding to the partially purified receptor was about a thousandfold more sensitive to DTNB than binding to this membrane-bound receptor. Dithiothreitol, a sulfide bond reducing agent, reversed the effects of DTNB on the opiate binding. PMID- 2989431 TI - Renin and angiotensin-converting enzyme in human neuroblastoma tissue. AB - High activity of renin was demonstrated in human neuroblastoma tissue. This activity was inhibited by specific antibody raised against human renal renin, indicating that it was not due to the nonspecific action of proteases. The specific activity of renin was 122.8 ng of angiotensin I generated mg of protein 1 h-1. It shared some biochemical features with well-known kidney renin, such as molecular weight, optimum pH, the presence of trypsin-activatable inactive renin, and glycoprotein nature. Furthermore, angiotensin-converting enzyme (ACE) activity (2.64 nmol mg of protein-1 min-1) was found in the tissue. This activity was inhibited by captopril, a specific ACE inhibitor, or by omission of chloride ion. These results suggest that true renin in addition to ACE exists in human neuroblastoma tissue. PMID- 2989432 TI - Molecular weight of human brain neutral sphingomyelinase determined in situ by the radiation inactivation method. AB - The radiation inactivation method was used to determine the molecular weight of membrane-bound neutral sphingomyelinase from normal human brain. Inactivation curves showed a molecular mass of 167,000 +/- 32,000. Molecular weights of two control enzymes, beta-N-acetylglucosaminidase and nonspecific beta-glucosidase, determined by the same procedure, were consistent with previous reports. PMID- 2989434 TI - Medium isotope effect in [3H]diazepam binding to benzodiazepine receptors of synaptic membranes. AB - Replacement of H2O by D2O resulted in significantly higher amount of [3H]diazepam specifically bound to synaptic membranes. The isotope effect arises from increased number of binding sites in D2O and is associated with a stronger solvation of membrane receptors by heavy water. PMID- 2989433 TI - Inhibition of calmodulin-stimulated phosphodiesterase activity by vasoactive intestinal peptide. AB - The effects of certain peptides of the glucagon family on calmodulin activity were determined from their capacity to inhibit a calmodulin-dependent form of phosphodiesterase. Vasoactive intestinal peptide and secretin were potent inhibitors of calmodulin activity, having IC50 values of 0.5 microM and 2 microM, respectively. By contrast, glucagon failed to inhibit calmodulin activity even at concentrations of 100 microM. None of these compounds significantly inhibited the basal activity of phosphodiesterase at concentrations up to 100 microM. These findings support the suggestion that important structural features of peptides for anticalmodulin activity include a net positive charge and a hydrophobic surface. PMID- 2989435 TI - Circulating T cell subsets in the Lambert-Eaton myasthenic syndrome. AB - Peripheral blood T cell subsets were measured using monoclonal antibodies and a fluorescence activated cell sorter in 15 untreated patients with Lambert-Eaton myasthenic syndrome (nine with small cell carcinoma, one undifferentiated epithelial tumour (ca-LEMS], five with no demonstrable tumour (non-ca-LEMS), 10 age-matched healthy controls and 10 patients with small cell carcinoma without neurological disease. OKT8+ (suppressor/cytotoxic) T cells were significantly decreased in ca-LEMS compared with non-ca LEMS (p less than 0.001) ca-controls (p less than 0.01) and healthy controls (p less than 0.001). In one patient depressed OKT8+ T cells antedated clinically evident tumour by five months. OKT3+ (total) and OKT4+ (helper) T cells were similar in ca-LEMS, non-ca LEMS and controls. The mechanism underlying the loss of circulating OKT8+ T cells in ca LEMS is unknown, but these changes may help to predict the presence of carcinoma in this disease. PMID- 2989436 TI - Peripheral neuropathy during longterm high-dose amiodarone therapy. AB - Three patients developed peripheral neuropathy after taking amiodarone for more than 18 months. All had high serum concentrations of amiodarone and its desethyl metabolite; in one patient concentrations in a sural nerve biopsy were 80 times higher than in serum. Peripheral neuropathy is a complication of large doses of amiodarone taken over long periods. PMID- 2989437 TI - Brachial plexus myoclonus. AB - Rhythmic myoclonus in an arm began abruptly following an injury and persisted continuously for six years. Topographical EMG showed abnormal activity confined to muscles innervated by the axillary and radial nerves from the posterior cord of the brachial plexus. Abduction of the arm above horizontal level stopped myoclonus and EMG discharges. EEG was normal. It is suggested that the myoclonus was caused by mechanical irritation of the posterior cord of the brachial plexus. PMID- 2989438 TI - Evidence on hypomyelination of central nervous system in murine muscular dystrophy. AB - To elucidate the disturbances of myelin metabolism in the nerve tissue of murine muscular dystrophy, the lipid composition of and the developmental changes in 2', 3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) and cholesterol ester hydrolase (CEHase) activities in the purified CNS myelin of dystrophic mice were determined. Several kinds of lipids, total galactolipid and cerebroside sulfatide levels were significantly reduced as compared with controls. Total cholesterol levels in the spinal cord of dystrophic mice were moderately higher. CEHase and, to a lesser degree, CNPase activities were reduced in the purified myelin of the CNS of the dystrophic mice. The reduced myelin CEHase activity in dystrophic mice suggests that impairment of hydrolysis of steryl esters may be important in the development of hypomyelination of the CNS. PMID- 2989439 TI - Changes in electrical threshold in human peripheral neuropathy. AB - A new method for analysis of excitability changes in human peripheral neuropathy is described. The range of the electrical threshold in a nerve is estimated in relative units from the ratio (S90-S10)/S10, where S10 and S90 are the stimulus voltages for 10% and 90% of maximal compound muscle potential. The ratio is increased if there is a fraction of high threshold fibres in the nerve; it is not affected by extraneural factors (e.g. distance to the nerve). The ratio was 0.16 +/- 0.03 in normals and significantly higher in subjects with chronic renal insufficiency or diabetes mellitus. Some of these had normal nerve conduction velocity. The ratio was markedly increased in the median nerve at the wrist in subjects with the carpal tunnel syndrome, which indicated that there are retrograde changes in the entrapped nerve fibres. PMID- 2989440 TI - Brain myelin of genetically obese mice. AB - Brain myelin was studied in genetically obese (ob/ob) mice and compared with that of normal (+/+) mice from the same strain. The brain from obese mice had a significantly lower amount of myelin, and marked changes in the fatty acid composition of myelin were observed. In contrast, the myelin cholesterol: phospholipid: galactolipid and the cerebroside: sulfatide molar ratios and also the phospholipid composition were normal. 2',3'-Cyclic nucleotide 3' phosphohydrolase (CNP) and UDP-galactose ceramide galactosyltransferase (CGalT) enzymatic activities were normal, and 3'-phosphoadenosine 5'-phosphosulfate (PAPS): cerebroside sulfotransferase (CST) was only slightly decreased. Cholesterol esters were not present in the mutant. These results indicate that brain myelination is affected in obese mutant mice. PMID- 2989441 TI - Focal deficiency of cytochrome c oxidase and of mitochondrial ATPase with histochemical evidence of loosely coupled oxidative phosphorylation in a mitochondrial myopathy of a patient with bilateral ptosis. An enzyme histochemical, immunocytochemical and fine structural study. AB - In the skeletal muscle of a patient with bilateral ptosis suggestive of progressive external ophthalmoplegia (PEO), but without ragged red fibres, electron microscopy revealed a moderate proliferation of mitochondria in nearly all fibres. A focal absence of cytochrome c oxidase and of mitochondrial ATPase was demonstrated histochemically in 3.2% and 1.4% respectively of the fibres. In 0.9% of the fibres both enzymes were deficient. In addition, mitochondrial ATPase, the ATP-synthesizing enzyme latent in controls, showed activation already before addition of an uncoupler. This indicates loosely coupled oxidative phosphorylation. The findings point to a complex derangement of mitochondrial function. Immunocytochemistry of cytochrome c oxidase favours the assumption that the defect is based on a highly diminished content of immunoreactive enzyme protein. PMID- 2989442 TI - Modulation of antigenic expression in cultured adult human oligodendrocytes by derivatives of adenosine 3',5'-cyclic monophosphate. AB - Oligodendrocytes were isolated from adult human brains obtained at autopsy by enzyme treatment - Percoll density gradient centrifugation, and grown in culture. During the first week in vitro, these cultures consisted of an enriched population (93-98%) of galactocerebroside-immunoreactive oligodendrocytes. After 2 weeks and onward, a large number of GFAP-positive astrocytes and glial cells doubly positive for galactocerebroside and GFAP markers was found among the oligodendrocytes. When these cultures were exposed to dibutyryl cyclic AMP, 8 bromocyclic AMP and RO-1724, an inhibitor of cyclic nucleotide phosphodiesterase, for 4-14 days, the majority of cells returned to express oligodendrocytic phenotype. These findings suggest the presence of heretofore unidentified "transitional" or "bipotential" glial cells in human brains that express both oligodendrocytic and astrocytic phenotypes, and the regulatory role of cyclic AMP derivatives which may induce a stable antigen expression in oligodendrocytes. PMID- 2989444 TI - Differential vulnerability of mixed and cutaneous nerves in lead neuropathy. AB - The prevalence of demyelinated fibers in mixed nerve (sciatic) and cutaneous nerve (sural) and the change in lead levels in various tissues over time were assessed in a model of lead neuropathy in the rat. Long-Evans rats were given drinking water containing 4% lead acetate and killed between one and 213 days of exposure. Lead levels in blood, brain, kidney, and femur increased over the 213 day period. Lead levels in sciatic nerve appeared to increase rapidly during the first few weeks of exposure and then decline to a lower plateau. The neuropathy was characterized by segmental demyelination and remyelination; neither axonal degeneration nor a microangiopathy was found. Sciatic nerve had a significantly greater prevalence of demyelinated fibers than sural nerve; the prevalence of demyelinated fibers was similar in proximal and distal sciatic nerve. The variable, brain-lead concentration times days on lead, which is an indicator of cumulative brain exposure, was the best predictor of the prevalence of demyelination. The differential involvement of sciatic and sural nerves in lead neuropathy may either indicate that Schwann cells myelinating different nerve fiber populations have different susceptibilities to lead toxicity, or that lead preferentially enters sciatic nerve. PMID- 2989443 TI - Phosphorylation of the sarcolemma of dystrophic and normal hamster muscle following denervation. AB - The rapid phosphorylation at 0 degree C of sarcolemma preparations of hamster leg muscle was compared with (Na+,K+)-ATPase activity in sham-operated and 7-day denervated muscle. The phosphorylation appeared to be under the trophic influence of the sciatic nerve since the denervated preparations exhibited a markedly reduced phosphorylation. In similar studies using dystrophic hamsters the sarcolemma preparations from sham-operated and denervated leg muscle both exhibited the same degree of phosphorylation. PMID- 2989445 TI - Treatment of nasopharyngeal carcinoma with human leukocyte interferon. AB - Nasopharyngeal carcinoma (NPC) is a human neoplasm closely associated with Epstein-Barr virus (EBV). Human leukocyte interferon (IFN) has known antiviral and antineoplastic properties. After initial IFN treatment in one NPC patient demonstrated acceptably low toxicity, 12 additional patients were treated on a protocol with IFN, 10 X 10(6) units intramuscularly (IM) daily for 30 days. IFN did not affect serum anti-EBV antibody titers (IgA and IgG antiviral capsid and early antigens). Of six patients tested, none was found to excrete EBV in saliva before, during, or after IFN. Four patients had measurable tumor regression (two partial responses and two minor responses), three had stable disease, and five patients plus the initial preprotocol patient had progressive disease. Toxicity included fever, fatigue, and myalgias in all patients, thrombocytopenia in two patients, and neutropenia in three patients. Three patients were withdrawn from the study, one each for severe fatigue, neutropenia, and hypotension. This study demonstrates that IFN has sufficient activity in advanced NPC to justify further investigation. PMID- 2989446 TI - Population-based study of small-cell lung cancer. PMID- 2989447 TI - Leukemia following treatment of germ cell tumors in men. PMID- 2989448 TI - Axonal projection patterns of ventrolateral medullospinal sympathoexcitatory neurons. AB - We studied the following properties of cat ventrolateral medullary (VLM) neurons that projected to the thoracic spinal cord: the relationship between their spontaneous activity and that in the inferior cardiac postganglionic sympathetic nerve, their responses to baroreceptor-reflex activation, their axonal conduction velocities, the funicular trajectories of their axons, the likely sites of termination of their axons, and their axonal branching patterns. Microstimulation in the second thoracic spinal segment (T2) antidromically activated 67 VLM neurons (as determined with time-controlled collision of spontaneous and evoked action potentials), whose activity was correlated to inferior cardiac sympathetic nerve discharge (as determined with spike-triggered averaging). We tested the effect of baroreceptor-reflex activation on the firing rate of 20 of these VLM spinal neurons. Because the firing rate decreased in each instance, these neurons apparently subserved a sympathoexcitatory function. The axonal branching patterns of 51 VLM-spinal sympathoexcitatory neurons were studied. Thirty-four neurons were antidromically activated by stimulation in the T2 gray matter and in more caudal thoracic spinal segments (T11 and/or T6). In each case, the antidromic response evoked by stimulation in the T2 gray matter was due to activation of an axonal branch rather than the main axon (via current spread to the white matter). This was demonstrated with tests that included time-controlled collision of the action potentials initiated by stimulation in T2 and a more caudal thoracic spinal segment. Some VLM-spinal axons that projected to T11 branched in T6 as well as in T2. These data indicate that some VLM-spinal neurons exerted widespread excitatory influences on sympathetic outflow. Seventeen VLM sympathoexcitatory neurons that innervated the T2 gray matter could not be antidromically activated by stimulation in T5, T6, and T11 despite an extensive search at each level. Thus the axonal projections of some VLM-spinal neurons were restricted to upper thoracic segments. Antidromic mapping in T2 revealed that the axons of VLM sympathoexcitatory neurons coursed through the dorsolateral or ventrolateral funiculus to innervate the region of the intermediolateral nucleus. Mean axonal conduction velocity was 3.5 +/- 0.3 m/s. Those VLM-spinal axons restricted to upper thoracic segments generally were located dorsally and/or medially to those that innervated widely separated thoracic segments. The discharges of 35 other VLM neurons that were antidromically activated by T2 stimulation were not related to sympathetic nerve activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2989449 TI - Axonal projections of single bulbospinal inspiratory neurons revealed by spike triggered averaging and antidromic activation. AB - Activity was recorded extracellularly from 26 inspiratory bulbospinal neurons in anesthetized, paralyzed, artificially ventilated cats. All but one were located in the ventral respiratory group. A neuron was classified as either I alpha or I beta by comparing its firing pattern during inspiratory cycles with lung inflation to its pattern when lung inflation was withheld during the central inspiratory phase (2, 14, 15). In this study, the projection and conduction velocity of these axons were determined using two methods: antidromic activation (AA) of the bulbospinal neurons and spike-triggered averaging (STA) of the extracellular field potentials. These methods have been compared directly because the same electrode was used both for stimulating the axon of the bulbospinal neuron and recording its axonal potential in the same location. Axonal projections from these neurons were mapped in the contralateral spinal cord with a mobile electrode by determining where the lowest stimulus threshold occurs for AA and greatest axonal potential can be recorded with STA. The locations of these axons were in the ventral and lateral funiculi. Each method determined a similar location for an axon. Positions of 10 axons were determined at both the third (C3) and fourth (C4) cervical segments. Single axons maintained their positions in either descending tract from rostral C3 to mid-C4. In five of six cases where two "neighboring" medullary units were characterized, the axons of each pair projected together within 350 micron of each other in the cervical spinal cord. Estimates of mean axonal conduction velocity (CV) from antidromic activation from a single stimulus site, "single-point AA," were as much as 42% less than corresponding estimates from STA extracellular field potentials at that point (P less than 0.001). Such single-point estimates were less than determinations that were calculated from the difference in conduction time and the difference in conduction distance from two points in the spinal cord. These two-point determinations averaged 55.4 +/- 13.1 m/s (using AA) and 53.3 +/- 13.1 (using STA) for 10 neurons. These values were not significantly (P greater than 0.2) different from each other and are greater than most earlier reports, which used the single-point AA method. Either method, AA or STA, can be used to determine axonal position and CV. The advantages and disadvantages of each method are discussed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2989450 TI - Dipeptides of glutamate and aspartate may be endogenous neuroexcitants in the rat hippocampal slice. AB - The dipeptide N-acetylaspartylglutamate (NAAG), and possibly the related dipeptide aspartylglutamate (AG), have been found in high concentrations in rat brain, and have been shown to bind avidly and selectively to a subset of glutamate (GLUT) receptors. Certain observations regarding GLUT and aspartate might be explained if the endogenous transmitter were a compound composed of both amino acids. We therefore examined the electrophysiological actions of NAAG and AG in the rat in vitro rat hippocampal slice model. NAAG or AG and GLUT were applied locally to cells by a dual-barrel pressure technique. Intracellular recordings from 34 CA1 pyramidal neurons showed depolarizations and conductance increases resembling evoked excitatory postsynaptic potential in 15 of 20 cells exposed to NAAG, and 14 of 14 exposed to AG. Many GLUT-responsive sites did not respond to AG, and most did not respond to NAAG. Responses to NAAG were usually too small to induce cell firing; they were best detected, therefore, by intracellular recording. With extracellular unit recordings, GLUT was equally excitatory in stratum radiatum and pyramidale of CA1 (N = 19; p greater than 0.10, one-way analysis of variance). In contrast, AG was considerably more potent (N = 21; p less than 0.01) in stratum radiatum. NAAG was not noted to excite cells when applied to stratum pyramidale. The region of maximal responsiveness to AG in CA1 coincided with the area of the dendritic tree receiving Schaffer collateral-commissural afferents. This spatial profile, together with other neuropharmacological evidence, support the candidacy of glutamate-containing peptides as endogenous excitatory compounds in certain pathways of hippocampus. PMID- 2989452 TI - Concussive head injury producing suppression of sensory transmission within the lumbar spinal cord in cats. AB - This study examines the effects of concussive levels of a fluid-percussion head injury on sensory transmission within the lumbar spinal cord of the cat. Primary afferent depolarization (PAD) was suppressed for 2 to 5 minutes following injury, as assessed by dorsal root potentials and augmentation of antidromic dorsal root potentials, both evoked by stimulation of adjacent dorsal roots. Polysynaptic reflex discharges in ventral root potentials evoked by dorsal root stimulation were also profoundly suppressed during this same period, even when spontaneous and monosynaptic reflex discharges were facilitated. Changes in PAD produced by injury were abolished by spinal cord transection, but were not affected by midpontine transection. These findings suggest that concussive head injury can produce suppression of segmental sensory transmission by neurally mediated processes involving the bulbar brain stem. Recordings of dorsal root resting potentials, antidromic dorsal root potentials, and reductions of antidromic dorsal root potentials induced by tetanic root stimulation indicated that depressed segmental sensory function produced by injury was due to suppression of postsynaptic interneuronal transmission rather than to excitability changes in primary afferent fibers. Somatosensory cortical potentials evoked by dorsal root stimulation were profoundly depressed at the same time as segmental sensory transmission was suppressed, suggesting that suppressed segmental sensory transmission may also contribute to suppression of ascending sensory transmission. It is hypothesized that transmission failure of interneuronal systems in the initial period following insult may be a general response occurring in wide areas of the central nervous system, and not restricted to areas to which mechanical stress is directly applied. This response pattern may result from indiscriminate activation of interconnected excitatory and inhibitory elements of interneuronal systems. PMID- 2989451 TI - The co-occurrence of a substance P-like peptide and cholecystokinin-8 in a fiber system of turtle cortex. AB - Single-label and double-label immunohistochemical techniques were used to demonstrate the coexistence of substance P-like immunoreactivity (SPLI) and cholecystokinin-8-like immunoreactivity (CCK-8-LI) in an extensive fiber system within the telencephalic cortex of turtle. All SPLI-containing fibers and terminals of this system contain CCK-8-LI and vice versa. The fibers of this system course from more medial cortical regions to more lateral ones, originating either from neurons in the more medial cortices or from extracortical neurons, the axons of which ascend the medial wall of the cortex. The precise location of the neurons that give rise to this cortical projection system is uncertain, but a hypothalamic location seems most likely at present. The fibers and terminals of this system are found throughout the entire mediolateral and rostrocaudal extent of the telencephalic cortex of turtle and are largely confined to the cell body layer of the cortex. Fewer SPLI/CCK-8-LI-containing fibers are found in pyriform (olfactory) cortex than in the other cortices. Ultrastructural studies indicate that SPLI/CCK-8-LI-containing terminals make asymmetric synapses on cell bodies or their proximal dendrites. Both SPLI and CCK-8-LI are found in large dense core vesicles in these labeled terminals. Labeled terminals also contained numerous small, round, unlabeled vesicles clustered near synaptic release sites and a number of unlabeled large dense core vesicles. Quantification of the percentage of the large dense core vesicles that were labeled in SP-labeled terminals, in CCK-8-labeled terminals, and in terminals labeled for both SP and CCK-8 provided suggestive evidence that SPLI and CCK-8-LI must be contained within the same large dense core vesicles. Radioimmunoassay indicated that the SP/CCK-8 containing system of turtle cortex contains 0.93 +/- 0.090 pg of SP/microgram of cortical tissue protein and 0.31 +/- 0.11 pg of CCK-8/micrograms of cortical tissue protein. The CCK-8-like material in turtle cortex coelutes with CCK-8 sulfate, using gradient elution high pressure liquid chromatography (HPLC). The SP-like material, although immunologically highly similar to undecapeptide SP (Reiner, A., J. E. Krause, K. T. Keyser, W. D. Eldred, and J. F. McKelvy (1984) J. Comp. Neurol. 226: 50-75), does not coelute with undecapeptide SP using gradient elution HPLC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2989453 TI - Comparison of fully automated and manual ejection fraction calculations: validation and pitfalls. AB - Resting multigated blood-pool studies were performed on 61 patients without arrhythmias and data were simultaneously acquired to two computer systems. Using one computer, manual ejection fraction (EF) was calculated by two trained observers. EF was also calculated from the other computer using a commercially available fully automated program; quality control (QC) images were routinely obtained to evaluate correct left-ventricular center location, background region assignment, or gross edge mispositioning. When errors were noted, the automated analysis was reprocessed with operator intervention. Forty-eight of the 61 studies (78%) produced adequate QC images. Operator redefinition of the left ventricular center and background of the 13 QC failures raised the automated success level to 92%. Correlation of the manual EF by two observers was excellent (r = 0.969). The automated EF measurements correlated well with the average observer's (r = 0.898). An improved version of software reduced the QC failures from 13 to 10. Operator assisted automated processing gave the success rate of 94%. The remaining 6% of patients required manual processing to obtain a correct ejection fraction. Our normal range for manual EF is greater than or equal to 0.50 and our corresponding normal range for this group of patients using the automated program was greater than or equal to 0.44. Based on greater than or equal to 0.44, four patients with a low manual EF had a normal automated EF. All four patients had cardiac disease but there was no evidence of abnormal cardiac function. The fully automated program provided good correlation with manual EF and can remove some of the subjectivity in manual edge determination. However, QC images must be carefully examined and the normal EF range for the automated program must be determined. PMID- 2989454 TI - A test of Cohen's developmental model for professional socialization with baccalaureate nursing students. AB - The study was designed to test the Cohen (1981) model of professional socialization, which proposes that nursing students progress through four developmental stages of unilateral dependence, negative/independence, dependence/ mutuality, and interdependence as they advance through an educational program. The Professional Socialization Staging Scale (PS3), consisting of four subscale scores representative of the stages, was administered to 422 students enrolled in eight clinical nursing courses. Instrument revision was accomplished using a cross-validation design, and the study hypotheses were tested with 214 subjects in the cross-validation subsample. Significant findings (p less than .05) were the beginning students were more dependent than were graduating students, that older students were less dependent and more highly interdependent than were younger students, and that students with concurrent work experience in nursing related fields were more highly interdependent than were students without concurrent work experience. With all groups of students evidencing the interdependence stage, no relationship was found between the sequential developmental stages and level of enrollment. Thus results of the study failed to support the Cohen model (McCain, 1984). PMID- 2989455 TI - A new look at nursing school program development: self- and external evaluation. AB - Developing and evaluating educational programs in schools of nursing is one of the most critical functions of the faculty. This process can be approached from a variety of perspectives. Historically, most faculty efforts have been directed at development or evaluation; however, a more proactive approach is to incorporate a process which will encompass both. To accomplish this, two models, the Holistic Formative Evaluation Model and the Stakes Countenance Model, have been merged to produce a model which is applicable to the integrated development and evaluation of a nursing curriculum. This model, the Internal Program Development and Self Evaluation Model, provides faculty, curriculum developers, and administrators with a comprehensive process which focuses on the monitoring program internal consistency and congruence. PMID- 2989456 TI - The success of non-BSNs in graduate nursing programs. AB - Although there is a great need to increase the number of master's prepared nurses, admissions criteria often limit the pool of possible applicants to those with a bachelor's degree in nursing (BSN), and/or require that non-BSN applicants complete special requirements to compensate for their lack of a degree in nursing. One of the few schools that admits RNs without the BSN, and does not require make-up of undergraduate nursing courses, is Yale University School of Nursing (YSN). Yale also has a program for preparing college graduates at the master's level for entry into nursing practice. Those individuals spend the first of three years in prespecialty basic nursing and the last two in the regular master's program. To determine if there was any difference in success among the groups at YSN, the three groups of individuals admitted to the two-year master's program at Yale--Non-Nurse College Graduates, BSNs, and nurses with a non-nursing baccalaureate degree were compared on their theoretical and clinical grades at the end of the first and second years of the specialty programs. Data were gathered from 435 students who had attended Yale over a ten-year period. No significant differences were found among these groups on clinical or theoretical grades at either the end of the first or second year of the program. This study indicates that although careful selection of candidates for graduate study is essential, intelligent, motivated individuals are able to acquire skills through self-learning and professional experience that are usually provided in baccalaureate nursing programs. PMID- 2989457 TI - The relationship of selected admission criteria to the academic success of associate degree nursing students. PMID- 2989458 TI - The community assessment assignment: process or product. PMID- 2989459 TI - Faculty practice policy. PMID- 2989460 TI - Designing learning to increase competency in interpersonal communication skills. PMID- 2989461 TI - Somewhere in the land of curriculum revision. PMID- 2989462 TI - Effect of wheat bran, pectin and cellulose on the secretion of bile lipids in rats. AB - We assayed the lipid content of bile from rats that had been fed either a standard diet (5% fat) or a high fat diet (25% fat, 1.2% cholesterol) in the presence or in the absence of various dietary fibers (namely, wheat bran, pectin and cellulose). The cholesterol concentration in bile from rats fed the high fat diet plus wheat bran or pectin was lower than that of the rats fed the high fat, high cholesterol diet without fiber. Bile phospholipids did not vary significantly from one group to another. In comparison to the standard diet, the high fat, high cholesterol diet led to a greater ratio of primary to secondary bile salts and a higher level of glycoconjugates. The observed differences may be explained by a variation in the metabolism of bile salts brought about by the difference in diet. PMID- 2989463 TI - Adenine nucleotide and lactate levels in organs from copper-deficient mice and brindled mice. AB - Experiments were conducted with 12-d-old suckling male mice to investigate energy metabolism during copper deficiency. Brindled mice, which have a genetic defect that affects copper distribution, were compared to their normal brothers and to nonbrindled mice that were copper deficient (-Cu) because their dams were consuming a diet low in copper since impregnation and to a fourth group of control suckling mice (+Cu) from copper-supplemented dams. A second study was done with older mice. Dietary copper deficiency was initiated at birth through dams and continued for 7 wk. Organs were fast-frozen in liquid nitrogen and then analyzed for adenine nucleotides by high performance liquid chromatography and for lactate spectrophotometrically. Levels of ATP, ADP and AMP were equivalent for liver, brain, heart and kidney from the young mice regardless of their copper status. Nucleotide levels were low in spleens from brindled mice. Lactate concentrations were elevated in brains from both -Cu and brindled mice but not in other organs. Older -Cu mice, in comparison, did have lower ATP levels and "energy charge" in heart but not liver compared to +Cu mice. Lactate levels were not higher despite a much lower cytochrome c oxidase activity. Thus, organs from copper-deficient mice with major reductions in cytochrome c oxidase activity do not necessarily have altered steady-state levels of adenine nucleotides. The reduction in cytochrome c oxidase activity in copper deficiency may play a minor role in the expression of pathophysiology. PMID- 2989464 TI - State and local health departments. Forgotten resources in occupational safety and health. PMID- 2989465 TI - Gingival metastasis from breast carcinoma. PMID- 2989466 TI - Use of collagen tubes for implantation of hydroxylapatite: an experimental study. AB - Collagen tubes were compared with routine injection for placement of hydroxylapatite in mandibular defects. It was found that the collagen film containers were effective in providing easier handling, more effective shaping, and less migration of particles. PMID- 2989467 TI - Sex hormone involvement in the development of experimental virally induced murine salivary gland tumors. AB - Mice were given a single inoculation of polyoma virus at birth and then orchidectomised or oophorectomised. Unoperated groups received polyoma virus alone whilst further males received testosterone, and females received oestradiol thrice-weekly in addition to polyoma virus inoculation. All groups were then observed over the succeeding 370 days for the development of tumors of the submandibular and parotid salivary glands. Polyoma virus inoculation selectively induced salivary gland tumors in 70% of male animals and 30% of female animals given polyoma virus alone. Testosterone therapy increased the salivary tumor frequency from 69-90% whilst orchidectomy reduced the tumor frequency to 50%. Oophorectomy or oestrogen therapy did not significantly alter the salivary gland tumor frequency in females but oestrogen therapy did result in the development of second primary tumors of breast in 60% of female animals bearing salivary gland tumors. The role of androgens in the development of virally induced salivary gland tumors is discussed as are the possible mechanisms responsible for the development of second primary tumor of breast. PMID- 2989468 TI - Morphological study of 391 cases of exocrine pancreatic tumours with special reference to the classification of exocrine pancreatic carcinoma. AB - Three hundred and ninety-one cases of primary pancreatic tumours, excluding endocrine tumours, were studied histologically. Carcinoma of the exocrine pancreas formed the largest group (98.5 per cent), benign tumours (1.25 per cent) and other malignant tumours (0.25 per cent) formed the remainder. Ductal adenocarcinoma was the commonest type and was divided into four sub-types, papillary, well, moderately and poorly differentiated duct adenocarcinoma. The moderately and poorly differentiated tumours were the commonest types. Papillary carcinoma was separated from the well differentiated tumours by its different morphological appearances and was found to exhibit different behaviour. Other special morphological types of pancreatic carcinoma, pleomorphic, mucinous, adenosquamous, acinar, microadenocarcinoma, cystadenocarcinoma and oncocytic carcinoma were also represented. Benign microcystadenomata (four cases) were considered because of their interesting morphological features and their significance in the differential diagnosis of carcinoma. Based on the morphology and behaviour of these 391 tumours, the classification of pancreatic carcinoma is discussed and some rare types are compared with previously reported cases and discussed. PMID- 2989469 TI - Immunohistochemical demonstration of S-100 protein in salivary gland neoplasms. AB - A peroxidase-antiperoxidase technique for S-100 protein has been applied to 68 salivary glands. These included 17 pleomorphic adenomas, seven adenoid cystic carcinomas, 23 adenolymphomas and a number of other neoplasms. In addition, five specimens of myoepithelial sialadenitis ('benign lymphoepithelial lesion') and five normal parotid glands were included. Consistent results were obtained, myoepithelial cells and cells in myxoid and chondroid areas in pleomorphic adenomas staining intensely. In adenoid cystic carcinoma, on the other hand, there was no staining. The adenolymphomas possessed intensely S-100 protein positive cells in the interfollicular lymphoid areas; these were probably interdigitating reticulum cells. In addition, branching structures, probably corresponding to Langerhans' cells, were observed in the epithelium of adenolymphomas. PMID- 2989470 TI - Immunohistochemistry of soft tissue tumours: a review. AB - The diagnosis and classification of soft tissue sarcomas can pose difficult problems for the histopathologist. Many sarcomas are too poorly differentiated to exhibit morphological features specific enough to define their histogenesis. Using the immunoperoxidase technique with commercially available antisera as a routine adjunct to other diagnostic aids, it is possible to arrive at more accurate diagnoses on which treatment protocols can be based. In addition a better understanding of mesenchymal neoplasms and their origins can be obtained by functional immunohistochemical studies. PMID- 2989471 TI - Human parvovirus (B19) and erythema infectiosum. AB - An outbreak of erythema infectiosum ("fifth disease") was studied in Fukuoka, Japan, in 1980-1981. Human parvovirus (HPV) antigen was not detected in any patients, but anti-HPV, measured by countercurrent immunoelectrophoresis, was found in 33 of 34 affected children and in 21 (15%) of 141 children of the same ages without the disease. Immunoglobulin M class anti-HPV was present in all 25 children with erythema infectiosum tested. In a survey of hospital patients, the prevalence of anti-HPV detected by CIE was 12% in the cohort 5 to 9 years of age, 19% in the cohort 10 to 14 years, and 32 to 55% in the cohorts greater than or equal to 30 years. The antibody reactions in the cases of erythema infectiosum, which were already well established at the onset of disease, indicate that HPV was the cause of the outbreak. PMID- 2989472 TI - Recurrence of Wilms tumor after apparent cure. AB - The recurrence of Wilms tumor after a 5-year disease-free interval is rare. We present two patients who had recurrent disease after a disease-free interval of greater than 7 years. Three additional patients, registered with the National Wilms Tumor Study who had a recurrence after 5 years, are also described. Of these three patients, two had nephroblastomatosis. Because more patients are achieving long-term survival, careful surveillance after apparent "cure" is recommended, particularly if nephroblastomatosis is identified in the original nephrectomy specimen. PMID- 2989473 TI - C-reactive protein in respiratory virus infections. PMID- 2989474 TI - Second-look laparotomy for Wilms' tumor: indications and results in 19 patients. AB - Nineteen of 74 children with Wilms' tumor underwent second-look laparotomy. Transperitoneal operation was done in five cases referred after flank operation elsewhere. Four had a change in stage from I to II or III, while one considered "inoperable" was resected (4/5 survived). Reoperation was done in two late referrals after operations with tumor spill. One had recurrent flank disease; the other had a flank mass with unrecognized diaphragmatic and intracaval extension (1/2 survived). Three children with giant tumors initially considered unresectable were successfully resected after cytoreduction with chemotherapy (3/3 survived). Two of three patients with bilateral Wilms' tumor survived reoperative procedures (partial or total nephrectomy). Five children with late intraabdominal recurrence (3 liver: 2 flank) eventually died despite reoperation and adjunctive therapy. All five had unfavorable histology. One child with en bloc hepatic resection had successful reoperation for suprahepatic vena caval obstruction due to regeneration of liver, but subsequently died. Ten of the 19 patients survived (52.6%) following reoperation and adjunctive therapy. Second look laparotomy is quite useful in patients inadequately staged with flank operations, in cases of bilateral Wilms' tumor, and in children with initially unresectable tumors following cytoreduction. Patients with extensive tumor spill at a previous procedure may benefit from early reoperation. Late recurrence of tumor (especially with unfavorable histology) and/or liver metastases carried an ominous prognosis. PMID- 2989475 TI - An evaluation of surgical treatment and chemotherapy of advanced neuroblastoma (stage III & IV) with special reference to proliferation kinetics of residual tumors. AB - Twenty two children with advanced retroperitoneal neuroblastoma and one child with advanced posterior mediastinal neuroblastoma admitted to our clinic were treated as follows. Seven patients (group A) underwent primary resection of tumor immediately after diagnosis. In two patients of this group, the levels of VMA and HVA in urine after surgery decreased to nearly normal (group A-I), while they did not change appreciably in the other 5 patients (group A-II). Seven patients (group B) underwent resection of tumor following complete or partial response to preoperative chemotherapy. Nine patients (group C) did not undergo resection of the tumor except for exploratory laparotomy. Two group A-I patients have survived, free of disease, for 6 months and 12 months after diagnosis. All patients of group A-II died within a year. Residual tumors of 4 patients of this group began to grow explosively just after surgery, although they received persistent postoperative chemotherapy. Four patients of group B survived for more than two years and the two patients of this group who received continuous intra arterial PGE1 therapy had no postoperative explosive growth of residual tumors. Two patients in group C survived for 20 months and the others died within a year. Primary tumors and metastatic foci responded well to therapy as compared with group A-II, which suggests that presence of primary tumors may inhibit rapid growth of metastatic foci. Resection of primary tumors, therefore, was not always conducive to survival unless residual tumor responded to postoperative chemotherapy. PMID- 2989476 TI - The prognostic implication of hypercholesterolemia in infants and children with hepatoblastoma. AB - This paper reports the relationship between serum cholesterol level and hepatoblastoma in nine patients. Four of the nine patients had a high (417-544 mg%) serum cholesterol. All of these patients were less than one year of age and had tumors of the epithelial type. Three of the infants died soon after being diagnosed. Three patients had a moderately elevated serum cholesterol (206 249mg%). One underwent primary hepatic resection. Two had nonresectable tumors that became resectable after chemotherapy. One died secondary to pulmonary metastases. The remaining two patients had a normal serum cholesterol level prior to treatment. Both patients had hepatic resection after chemotherapy and had no evidence of increased cholesterol postoperatively and are alive six years after diagnosis. From these data, it is suggested that the pretreatment level of serum cholesterol may be of prognostic significance in infants and children with hepatoblastoma. PMID- 2989477 TI - Freeze-dried bone and coralline implants compared in the dog. AB - Partially demineralized, antigen-extracted, freeze-dried cadaver bone and a hydroxyapatite replica of a marine coral skeleton were implanted in 30 surgically created two-wall intrabony pockets in two dogs. Results after 6 and 8 months showed connective tissue encapsulation and some periodontal ligament formation around the cadaver bone. In contrast, the coralline implant was consistently invaded by fibrovascular tissue, showed bone formation on its internal surfaces and had some periodontal ligament formation around it. PMID- 2989478 TI - [Preliminary study of nitrogen encapsulation on Zeolite 3A and its pharmaceutical application]. PMID- 2989479 TI - A high-performance liquid chromatographic method for the assay of Na+, K+ adenosine triphosphatase inhibition. AB - A new method for the assay of Na+, K+-adenosine triphosphatase (Na+, K+-ATPase) inhibition has been devised involving the determination of enzymatically produced adenosine diphosphate (ADP) and unchanged adenosine triphosphate (ATP) by high performance liquid chromatography (HPLC). The substrate, ATP, was incubated with the enzyme preparation in the presence of an inhibitor. The incubation mixture was filtered through a membrane filter, and ADP and ATP in the filtrate were separated by (HPLC). The inhibitory effect of a cardiac steroid on the enzymic reaction was estimated by measuring the peak area ratio of ADP to ADP plus ATP on the chromatogram. The proposed assay method has proved to be satisfactory with respects to simplicity, sensitivity, and reproducibility. PMID- 2989480 TI - Influence of nourseothricin on growth and secondary metabolism of Streptomyces noursei JA 3890b. AB - The nourseothricin-producing S. noursei strain JA 3890b possessed a high degree of resistance to its own antibiotic when grown in submerged cultures started from mycelium samples as inocula. In contrast, both the outgrowth of spores and the development of surface colonies from mycelium samples were severely inhibited in the presence of relatively low concentrations of nourseothricin, suggesting that the producer organism is susceptible to the autotoxic metabolite in particular stages of its development. Nourseothricin production by submerged cultures has been found to be independent of negative feedback regulation by the antibiotic. PMID- 2989481 TI - Localization of a streptothricin acetyl transferase in cells of Escherichia coli K-12. AB - Streptothricin acetyl transferase coded for by plasmids pIE636 and pIE637 in Escherichia coli K-12 was found to be located at the inner side of the cytoplasmic membrane. PMID- 2989482 TI - Synthesis and biological evaluation of the superagonist [N alpha chlorotriazinylaminofluorescein-Ser1,Nle4,D-Phe7]-al pha-MSH. AB - The fluorescein-labeled melanotropin [N alpha-chlorotriazinylaminofluorescein Ser1,Nle4,D-Phe 7]-alpha-MSH, was prepared by solid-phase techniques of peptide synthesis. The biological actions of this analogue were determined in several melanocyte bioassays and were compared with the parent peptide [Nle4,D-Phe7] alpha-MSH and the native hormone alpha-MSH. The fluorescein compound was a superpotent agonist with approximately 10 times more activity than alpha-MSH in both the frog and the lizard skin bioassays. Murine S91 melanoma cells assayed in vitro (tyrosinase bioassay) were as responsive to the fluorescein analogue as to alpha-MSH. The analogue exhibited ultraprolonged biological activity and the biological activities were unaffected by treatment of the analogue with alpha chymotrypsin. The fluorescein-labeled melanotropin should prove useful for melanotropin receptor characterization. PMID- 2989483 TI - Application of differential scanning calorimetry to the study of solid drug dispersions. AB - The present study describes the application of differential scanning calorimetry (DSC) to ascertain the crystalline state of a drug with a melting point of approximately 53 degrees C after dispersion on hydrophilic carriers by either simple mixing or by fusion. The carriers examined include polyethylene glycol 6000 and colloidal silicon dioxides. The most interesting of the systems investigated, in which the drug is gradually transformed from the crystalline to the amorphous state at room temperature, are physical mixtures of the drug and colloidal silicon dioxides. The crystalline transformation is manifested by the gradual decrease in the endothermic transition energy of the physical mixture with time. The crystalline transformation is characteristically biphasic with initially fast first-order kinetics, followed by a slow conversion process. The rate of transformation is dependent on the drug-silicon dioxide ratio, temperature, and certain physical properties of the silicon dioxides. An inverse relationship exists between transition energy and the in vitro dissolution rate of the drug in the physical mixtures with silicon dioxide. This suggests that DSC may provide a useful method for evaluating the effects of formulation variables upon dissolution rate. PMID- 2989484 TI - Simultaneous liquid chromatographic determination of glutaric acid, phenylephrine, and benzyl alcohol in a prototype nasal spray with application to di- and tricarboxylic acids. AB - A rapid reversed-phase high-performance liquid chromatographic method for the simultaneous determination of glutaric acid, phenylephrine, and benzyl alcohol in nasal spray has been developed. UV detection was utilized at 210 nm for the assay of glutaric acid and phenylephrine with an adjustment to 254 nm for the measurement of benzyl alcohol. Linearity and recovery data were obtained for each component in spiked placebo studies. An investigation of the retention mechanisms of the three components showed that phenylephrine was retained by ion-pairing with octanesulfonate anion while glutaric acid and benzyl alcohol partitioned as a suppressed ion and a neutral molecule, respectively. The method has been further extended to the reversed-phase separation of di- and tricarboxylic acids using a totally aqueous 0.0074 M phosphoric acid mobile phase. The retention of these acids was related to their octanol-water partition coefficients and structural variation. PMID- 2989485 TI - Synthesis of N2-[(S)-1-carboxy-3-phenylpropyl]-L-lysyl-L-proline (lisinopril). AB - The synthesis and some of the spectral properties of N2-[(S)-1-carboxy-3 phenylpropyl]-L-lysyl-L-proline (lisinopril, MK-521) are described. This compound inhibits angiotensin-converting enzyme with an IC50 of 1.2 X 10(-9) M. PMID- 2989486 TI - Behavioral studies with anxiolytic drugs. I. Interactions of the benzodiazepine antagonist Ro 15-1788 with chlordiazepoxide, pentobarbital and ethanol. AB - Lever pressing by squirrel monkeys was maintained under two behavioral procedures known to be sensitive to anxiolytic drugs. Under one procedure, responding maintained by food was suppressed by electric shock (punishment). Under a second procedure, responding was maintained under a multiple schedule in which the first response after 5 min produced either food or shock depending on the stimulus that was present throughout the interval (fixed-interval schedule). Under the punishment schedule, chlordiazepoxide (1.0-100 mg/kg), pentobarbital (1.0-17.0 mg/kg) and ethanol (0.5-2.5 g/kg) increased responding. The benzodiazepine antagonist, Ro 15-1788 (1.0-10.0 mg/kg), which was without behavioral activity when given alone, reversed the effects of chlordiazepoxide in a dose-dependent manner. Ro 15-1788 did not antagonize the effects of pentobarbital or ethanol but potentiated the rate-increasing effects of these compounds. Under the multiple fixed-interval food- or shock-presentation schedule, both chlordiazepoxide and pentobarbital increased responding maintained by food but only decreased responding maintained by shock. Ro 15-1788 antagonized the rate-increasing effects of chlordiazepoxide under the food schedule and reversed the rate decreasing effects during the shock-presentation schedule; pentobarbital effects were not altered by Ro 15-1788. Certain dose-combinations of chlordiazepoxide and Ro 15-1788 produced large increases in responding maintained by shock, an effect not seen with either drug alone. These studies indicate that Ro 15-1788 antagonizes the behavioral effects of benzodiazepines selectively but not those of other sedative-hypnotic drugs. These results also suggest that Ro 15-1788 may exert certain actions of its own or may unmask other drug effects when given in combination with benzodiazepine and nonbenzodiazepine compounds. PMID- 2989488 TI - Sympathetic denervation does not alter the density or properties of alpha-1 adrenergic receptors in rat vas deferens. AB - Alpha-1 adrenergic receptors in surgically denervated rat vas deferens were studied using radioligand binding assays of [125I] BE 2254 ([125I]BE) and contraction measurements. Scatchard analysis of saturation isotherms of specific [125I]BE binding showed no change in the affinity or density of binding sites 4, 7 or 14 days after denervation of rat vas deferens. The potency of norepinephrine in inhibiting specific [125I]BE binding was also unchanged 7 days after denervation of vas deferens. The potency of phenylephrine in causing contraction in vitro did not change 4, 7 or 14 days after denervation of vas deferens; however, there was a significant increase in the maximum contractile response to phenylephrine at all time points. After partial inactivation of alpha-1 adrenergic receptors in vitro with phenoxybenzamine, there was an equivalent reduction in the number of [125I]BE binding sites in the control and 14-day denervated vas deferens. The equilibrium dissociation constants calculated from contractile measurements for norepinephrine were the same in the control and denervated tissues. However, there was a 2.2-fold increase in contractile sensitivity to norepinephrine 14 days after denervation and a 3.6-fold increase in contractile sensitivity to methacholine 7 days after denervation. These results show that surgical denervation of the rat vas deferens results in an increase in contractile sensitivity to norepinephrine and methacholine and an increase in maximum contraction. However, there is no change in alpha-1 adrenergic receptor density or properties at any time after denervation. Thus, alterations in alpha-1 adrenergic receptors do not contribute to contractile supersensitivity of denervated rat vas deferens. PMID- 2989487 TI - Differences in responsiveness of intrapulmonary artery and vein to arachidonic acid: mechanism of arterial relaxation involves cyclic guanosine 3':5' monophosphate and cyclic adenosine 3':5'-monophosphate. AB - The objective of this study was to examine the relationship between responses of bovine intrapulmonary artery and vein to arachidonic acid and cyclic nucleotide levels in order to better understand the mechanism of relaxation elicited by arachidonic acid and acetylcholine. Arachidonic acid relaxed phenylephrine precontracted arterial rings and elevated both cyclic GMP and cyclic AMP levels in arteries with intact endothelium. In contrast, endothelium-damaged arterial rings contracted to arachidonic acid without demonstrating significant changes in cyclic nucleotide levels. Indomethacin partially inhibited endothelium-dependent relaxation and abolished cyclic AMP accumulation whereas methylene blue, a guanylate cyclase inhibitor, partially inhibited relaxation and abolished cyclic GMP accumulation in response to arachidonic acid. All vessel responses were blocked by a combination of the two inhibitors. Prostaglandin (PG) I2 relaxed arterial rings and elevated cyclic AMP levels whereas PGE2 and PGF2 alpha caused contraction, suggesting that the indomethacin-sensitive component of arachidonic acid-elicited relaxation is due to PGI2 formation and cyclic AMP accumulation. The methylene blue-sensitive component is attributed to an endothelium-dependent but cyclooxygenase-independent generation of a substance causing cyclic GMP accumulation. Intrapulmonary veins contracted to arachidonic acid with no changes in cyclic nucleotide levels and PGI2 was without effect. Homogenates of intrapulmonary artery and vein formed 6-keto-PGF1 alpha, PGF2 alpha and PGE2 from [14C]arachidonic acid, which was inhibited by indomethacin. Thus, bovine intrapulmonary vein may not possess receptors for PGI2. The failure of endothelium-intact vein to relax to acetylcholine may be related to the lack of a relaxant effect by arachidonic acid, perhaps attributed to the absence of generation of an endothelium-derived relaxing factor. PMID- 2989489 TI - Sigma opioid receptor: characterization and co-identity with the phencyclidine receptor. AB - The properties of the sigma opioid receptor of rat brain cortex have been characterized using the prototypic ligand (+)-[3H] SKF 10,047. Binding to this receptor was rapid, and equilibrium was obtained within 30 min at 37 degrees C. Specific binding was linear with protein concentration up to 500 micrograms/2 ml and was dependent upon protein integrity. Denaturation by boiling destroyed over 95% of the specific binding. A high-affinity binding site with a KD of 150 +/- 40 nM and a maximum binding of 2.91 +/- 0.84 pmol/mg of protein was determined from a Scatchard plot of the binding data. The addition of salt, either NaCl or CaCl2, to the buffers markedly decreased binding, with CaCl2 being more potent than NaCl. A broad pH optimum for specific binding was observed; maximum binding was at pH 9.0. The affinity of a number of ligands for the sigma site and the phencyclidine receptor were compared. The binding (IC50) of 13 ligands to the sigma site showed a correlation of 0.86 (P less than .01) with binding to the phencyclidine site. The data demonstrate that the biochemical properties of the sigma and phencyclidine receptors are similar and support the view that these receptors are one and the same site. PMID- 2989490 TI - Solubilization and characterization of putative alpha-2 adrenergic isoceptors from the human platelet and the rat cerebral cortex. AB - Alpha-2 adrenergic receptor isotypes have been proposed to explain several pharmacologic differences between rodent and nonrodent species. In support of this hypothesis, we found that the differences in the pharmacologic properties of rat cerebral cortex and human platelet alpha-2 adrenergic receptors were not due to 1) differential proteolysis of the receptor, 2) degradation of the ligand, 3) the detection of different affinity states or 4) the presence of different quantities of various affinity modulators. In an effort to test the hypothesis further we have characterized these prototype isoceptors in soluble form. Soluble receptors from both species showed the appropriate rank order of potencies for various adrenergic agonists and antagonists expected for an alpha-2 adrenergic receptor. The KD values for soluble human platelet and rat cerebral cortical alpha-2 receptors were 4.2 and 5.9 nM, respectively. The species differences in the affinities of prazosin and oxymetazoline were retained upon solubilization. Both soluble receptors were insensitive to modulation by guanine nucleotides, indicating uncoupling from the inhibitory regulatory subunit Ni. Sucrose density gradient centrifugation of soluble receptors did not indicate any significant molecular size differences. PMID- 2989491 TI - Calcium entry blockade by nitrendipine and alpha adrenergic responsiveness in vivo: comparison with noncalcium entry blocker vasodilators in absence and presence of phenoxybenzamine pretreatment. AB - To investigate the in vivo interaction between calcium entry blockade by nitrendipine (a dihydropyridine calcium entry blocker) and alpha adrenergic mediated end-organ responsiveness, four series of experiments were carried out in normal Sprague-Dawley rats. In ganglion-blocked rats (hexamethonium, 10 mg/kg i.p. plus atropine, 1.0 mg/kg i.p.), nitrendipine (0.3 mg/kg) antagonized the pressor responses to angiotensin II and vasopressin as well as to norepinephrine, thus indicating the lack of specificity of its antagonism to alpha adrenergic vasoconstriction. The results of the next two series of experiments showed first that, in pithed rats, nitrendipine (0.01 to 0.3 mg/kg) in presence of prazosin shifted the norepinephrine pressor dose-response curves to the right whereas it was ineffective in yohimbine-pretreated animals. These data, suggesting a preferential alpha-2 antagonism by nitrendipine, were confirmed further by its little effect on pressor responses to methoxamine as contrasted with its marked progressive depression of the maximum response to B-HT 920 (about 80% at the highest rate of infusion). However, qualitatively similar results were obtained by the noncalcium entry blocker vasodilators, both sodium nitroprusside and hydralazine, both of which led to minor shifts to the right of the methoxamine pressor dose-response curves, whereas dose-dependently depressing the maximum pressor response to B-HT 920 (about 70 and 40%, respectively). Thus, calcium entry blockade appeared to antagonize preferentially alpha-2-mediated vasoconstriction, but this effect was common to other vasodilators devoid of calcium entry blocking properties.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989492 TI - Characterization of [3H]leukotriene D4 binding sites in guinea-pig ventricular myocardium. AB - [3H]Leukotriene (LT) D4 was used to identify specific LTD4 binding sites in guinea-pig ventricular myocardial membranes. High-performance liquid chromatography analyses indicated that, in the presence of the gamma-glutamyl transpeptidase inhibitor L-serine-borate (80 mM), less than 3% of membrane-bound [3H]LTD4 was converted to [3H]LTC4 or [3H]LTE4 at 30 degrees C. The specific [3H] LTD4 binding, assayed in the presence of 80 mM L-serine-borate, reached a stable steady state within 45 min at 30 degrees C (pH 7.5). A monophasic Scatchard plot of saturation binding data yielded an apparent dissociation constant (Kd) of 3.4 +/- 2.1 nM and a maximum number of binding sites of 850 +/- 91 fmol/mg of protein. Competition binding studies with [3H]LTD4, synthetic 5S, 6R-LTD4 (LTD4) and its diastereoisomer 5R,6S-LTD4, LTE4, LTC4 and the putative LT antagonists FPL 55712, 4R-hydroxy-5S-1-cysteinylglycine-6Z-nonadecanoic acid (2-nor-LTD1) and SKF 88046 demonstrated a potency order of LTD4 greater than LTE4 greater than LTC4 greater than 5R,6S-LTD4 much greater than 2-nor-LTD1. FPL 55712 and SKF 88046 were ineffective in displacing the specific [3H]LTD4 binding. Pretreatment of the heart membranes with the sulfhydryl reducing reagent dithiothreitol decreased the specific [3H]LTD4 binding in a concentration-dependent manner. Scatchard analyses of saturation isotherms indicated that 0.3 mM dithiothreitol pretreatment of heart membranes decreased the maximum number of binding sites of the [3H]LTD4 binding to 368 +/- 61 fmol/mg of protein with minimal effects on the apparent Kd.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989493 TI - Aluminum and gallium arrest formation of cerebrospinal fluid by the mechanism of OH- depletion. AB - AlCl3 or GaCl3 was added to artificial cerebrospinal fluid and perfused through the cerebral ventricles of the rat. Depending on the metal and its concentration (1-10 mM) the pH of the perfusate ranged from 7.2 to 3.5. At 10 mM metal chloride, yielding pH 4.7 (Al) or 3.5 (Ga), formation of cerebrospinal fluid was suppressed 100%. The effect was reversed as soon as control cerebrospinal fluid (pH 7.35) was introduced. There was no effect at pH greater than 6 in the presence of metal ions nor was the effect mimicked by cerebrospinal fluid acidified with HCl or phosphate buffer to pH 4.7. HCl and phosphate at pH 4.7 had partial effects (25-30%). Inhibition of carbonic anhydrase is well known to have a partial effect (approximately 50%), the remainder of normal flow being dependent on the uncatalyzed formation of HCO3- and the corresponding movement of Na+. The complete cessation of flow after exposure to Al or Ga ions appears to occur when these ions are perfused at pH where they are partly hydrolyzed. Under these circumstances they form very powerful proton generating systems (as shown by titration data), which could lower [OH-] at the secretory border of choroid plexus cells. As a result, both the catalyzed and the uncatalyzed processes for formation of HCO3- from CO2 are abolished and secretion stops. This mechanism may also account for the antiperspirant action of Al salts. Using metal ion hydrolysis to probe other secretory systems will also be of interest. PMID- 2989494 TI - Opioid binding to rat and guinea-pig neural membranes in the presence of physiological cations at 37 degrees C. AB - We have identified mu, delta and kappa opioid binding sites in four types of neural membranes under conditions which include physiological concentrations of ions and an incubation temperature of 37 degrees C. We hypothesize that binding parameters determined under these conditions should be more directly comparable with physiological experiments than parameters obtained under conditions of low ionic concentration and at low temperature. By using either a radioligand which is selective for a single type of opioid binding site or a relatively nonselective radioligand in the presence of an unlabeled selective ligand, we have isolated binding to single populations of sites. Saturation and displacement data were analyzed with the aid of a computerized nonlinear curve fitting program. [3H]Tyr-D-Ala-Gly-(Me)Phe-Gly-ol bound to a single population of sites with the characteristics of mu receptors, as determined by saturation and displacement analysis. Binding to the mu site represented 70% of the total specific opioid binding in rat brain, but only 20 to 30% in guinea-pig tissues. [3H][D-Ala2-D-Leu5]enkephalin bound almost equally well to mu and delta sites, but the delta site could be examined by the inclusion of unlabeled Tyr-D-Ala-Gly (Me)Phe-Gly-ol in the incubations. [3H]Ethylketocyclazocine bound mu and kappa sites, and Tyr-D-Ala-Gly-(Me)Phe-Gly-ol was also used to block the mu component in experiments in which we studied kappa binding. Binding to kappa sites represented 50 to 60% of the total in guinea-pig tissues, but less than 20% in rat brain. PMID- 2989495 TI - In vivo receptor binding of opioid drugs at the mu site. AB - The in vivo receptor binding of a series of opioid drugs was investigated in intact rats after s.c. administration of [3H]etorphine tracer, which selectively binds to mu sites in vivo. Receptor binding was determined by a membrane filtration assay immediately after sacrifice of the animals and brain homogenization. Coadministration of unlabeled opioid drugs together with tracer led to a dose-dependent decrease of in vivo tracer binding. Estimates of the doses required to occupy 50% of the mu sites in vivo established the following potency rank order: diprenorphine, naloxone, buprenorphine, etorphine, levallorphan, cyclazocine, sufentanil, nalorphine, ethylketocyclazocine, ketocyclazocine, pentazocine, morphine. In vivo-in vitro differences among the relative receptor binding potencies were only partially accounted for by differences in their access to the brain and the regulatory effects of Na+ and GTP, which are expected to reduce agonist affinities in vivo. The relationship among mu receptor occupancy in vivo and pharmacological effects of the opioid drugs is described. PMID- 2989496 TI - Transport of p-aminohippurate, tetraethylammonium and D-glucose in renal brush border membranes from rats with acute renal failure. AB - Transport of D-glucose, p-aminohippurate and tetraethylammonium has been studied using renal brush border membrane vesicles isolated from rats with uranyl nitrate induced acute renal failure (ARF). Initial rate and overshoot magnitude of Na+ gradient-dependent D-glucose uptake were decreased in brush border membrane vesicles from ARF rats compared with normal rats, although there was no significant difference on D-glucose uptake in the presence of equilibrated Na+ between normal and ARF rats. Uptake of p-aminohippurate by membrane vesicles from ARF rats did not differ from normal membrane vesicles. Uptake of tetraethylammonium with or without an H+ gradient was decreased in membrane vesicles from ARF rats compared with normal rats. Dissipation rate of H+ gradient across brush border membranes did not differ between both groups. In vitro incubation of normal brush border membrane vesicles with uranyl nitrate caused no alteration in any substrate transport. However, enzyme activities such as (Na+ + K+)-adenosine triphosphatase in renal cortical homogenate were inhibited markedly in the presence of uranyl nitrate. These results suggest that uranyl nitrate induced ARF caused alterations in the transport properties of renal brush border membranes and that these transport dysfunctions were not due to the direct effect of uranyl nitrate, but could be secondarily induced after the impairment of the integrity for tubular cells. PMID- 2989497 TI - Inhibitory effects of norepinephrine, methoxamine and phenylephrine on renin release from rat kidney cortical slices. AB - We attempted to determine whether the inhibition of renin release from rat kidney cortical slices by alpha adrenoceptor agonists is mediated by activation of alpha 1 and/or alpha-2 adrenoceptors, and to investigate the role of calcium in the mechanisms of this inhibition. Norepinephrine (NE), methoxamine (ME) and phenylephrine (PE) produced a concentration-dependent inhibition of renin release from rat kidney cortical slices, whereas clonidine was without effect. NE-, ME- and PE-induced inhibition of renin release was blocked by prazosin, which was 2 or 3 orders of magnitude more potent than yohimbine. The inhibitory effects of NE, ME and PE on renin release from the slices were abolished by removal of calcium from the incubation medium. Calcium antagonists, verapamil and nifedipine, attenuated the responses of renin release to NE, ME and PE, in a concentration-dependent manner. The inhibitory effects of NE, ME and PE on renin release were blocked significantly by N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide, a calmodulin antagonist, but not by N-(6-aminohexyl)-1 naphthalenesulfonamide, which has virtually no calmodulin antagonistic activity. These findings suggest that alpha adrenoceptor agonists inhibit renin release from rat kidney cortical slices mainly via alpha-1 adrenoceptors and that calcium influx followed by the activation of the calcium-calmodulin system is involved in the above inhibition. PMID- 2989498 TI - Disposition of enalapril in the perfused rat intestine-liver preparation: absorption, metabolism and first-pass effect. AB - A new procedure, namely the in situ perfused rat intestine-liver preparation, was introduced to examine the roles of the intestine and the liver in the elimination of enalapril, a new angiotensin-converting enzyme inhibitor. The in situ perfused rat intestine preparation was used to determine the rate and extent of enalapril absorption after an-intraduodenal dose. In the former technique, enalapril in blood perfusate (10 ml/min) was delivered via the superior mesenteric artery into the once-through perfused rat intestine-liver preparation, with sampling effected in reservoir, portal vein and hepatic vein. The ease of sampling, proximal and distal to the intestine and liver, allowed the direct estimation of the extraction ratios by the intestine and the liver. The steady-state intestinal extraction ratio of enalapril was small (0.04 +/- 0.066) compared to that for the liver (0.74 +/- 0.06), indicating that the liver was responsible for most of the hydrolytic conversion of enalapril to its pharmacologically active diacid metabolite, enalaprilat. Moreover, no trend in the values of the extraction ratios by both organs was apparent among the input concentrations of enalapril (0.55, 2.6 and 13.3 microM) used. Portal venous plasma consisted mainly of enalapril and was devoid of enalaprilat, whereas both enalapril and enalaprilat were detected in bile and hepatic venous plasma. With the latter technique, an intraduodenal injection of a tracer dose of [14C]enalapril (0.14-0.39 mumol) was made close to the pyloric sphinctor, whereas the intestine preparation was recirculated (7.5 ml/min) with blank perfusate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989499 TI - CM 40907: a structurally novel anticonvulsant in mice, rats and baboons. AB - CM 40907 [3-(4-hydroxypiperidyl)-6-(2'-chlorophenyl)-pyridazine] is a chemically original compound which possesses the pharmacological properties of a potent, p.o. active anticonvulsant. The anticonvulsant activity of CM 40907 was examined in mice, rats and photosensitive Papio-papio baboons and compared to that of phenobarbital, diphenylhydantoin, carbamazepine, sodium valproate and ethosuximide. In mice, CM 40907 antagonized electroconvulsive shock and chemically induced seizures with an overall potency comparable to that of carbamazepine and a therapeutic ratio (ED50 rotorod/ED50 electroshock) superior to that of ethosuximide, sodium valproate, phenobarbital and carbamazepine. In the rat CM 40907 suppressed completed kindled amygdaloid seizures and was approximately as active as phenobarbital. In naturally photosensitive Senegalese Papio-papio baboons CM 40907 antagonized myoclonus and cortical paroxysmal discharges. In this model CM 40907 was approximately one-fourth as potent as phenobarbital, twice as potent as carbamazepine and 6 times more potent than sodium valproate. In mice CM 40907, at anticonvulsant doses, increased the affinity of [3H]flunitrazepam for its central receptor site. Based on these results it is postulated that CM 40907 is a potent and relatively nonsedative anticonvulsant and may be of therapeutic benefit in epileptic disorders. PMID- 2989500 TI - Plasma corticosterone changes in response to central or peripheral administration of kappa and sigma opiate agonists. AB - The acute administration of morphine produces a characteristic increase in plasma corticosterone. By using the male rat with a chronic i.v. catheter, a stereotaxically placed cannula in the lateral ventricle and a sound-attenuated one-way vision chamber, the effects of prototypic kappa and sigma opiate receptor agonists were studied. Both ethyl-ketocyclazocine (EKC) and N-allyl-normetazocine [SKF 10047] (SKF) also produced such a rise in hormone level after i.v. or i.c.v. administration. The former effect was blocked by concurrent treatment with naloxone (NX), 0.4 mg/kg. The latter effect was not blocked by i.c.v. pretreatment with beta-funaltrexamine, a long-acting mu receptor antagonist, whereas the response to i.c.v. morphine was attenuated significantly. The development of acute dependence and short-term tolerance to EKC and SKF was also studied. Priming with either drug (i.v.) did not result in a NX-precipitated plasma corticosterone withdrawal response 3 hr later. Similar studies priming with morphine (i.c.v.) did result in the plasma corticosterone-elevated response when NX was administered i.c.v. after 3 hr. When EKC or SKF was substituted for morphine, no NX-induced response was observed. Short-term tolerance to the effects of EKC and SKF on the hypothalamo-pituitary-adrenal axis did not appear to occur. These data support the notion that stimulation of several subclasses of opiate receptors will result in the activation of the hypothalamo-pituitary adrenal axis. Furthermore, it appears that the mu opiate receptor is involved in the initiation of acute opiate dependence. PMID- 2989501 TI - Desensitization of alpha-1 adrenergic receptor-mediated vascular smooth muscle contraction. AB - Desensitization of alpha-1 receptor-mediated smooth muscle contraction was studied in rabbit aorta. Incubation of rabbit aorta ring segments with epinephrine (10(-6) M) for 7 hr resulted in a 10-fold loss in sensitivity of the tissue to alpha-1 adrenergic receptor-mediated contraction with no change in maximal force of contraction. This loss in sensitivity was specific for alpha-1 receptor-mediated contraction because responses to histamine and serotonin were unchanged in these aortas. Conversely, prolonged exposure of vessels to histamine (10(-5) M) led to desensitization of histamine-mediated contraction without altering responses to alpha-1 receptor stimulation. Using [125I] BE2254, a potent alpha-1 receptor antagonist, the loss in sensitivity to catecholamines was found not to be mediated by down-regulation of alpha-1 receptors nor by a loss in their affinity for epinephrine. However, desensitization was associated with a blunting of alpha-1 receptor stimulation of phosphatidylinositol turnover. These results suggest that desensitization of alpha-1 receptor-mediated contraction in rabbit aorta does not appear to be mediated by changes in receptor number or affinity but may involve alterations in receptor coupling. PMID- 2989502 TI - Role of extracellular calcium in contractions produced by activation of postsynaptic alpha-2 adrenoceptors in the canine saphenous vein. AB - Canine saphenous vein (CSV) has been shown to contain both postsynaptic alpha-1 and alpha-2 adrenoceptors. Our previous studies have shown that activation of postsynaptic alpha-1 adrenoceptors in this tissue utilizes both extracellular and intracellular Ca++ to produce contractions. In the present study, the source of calcium mobilized by activation of postsynaptic alpha-2 adrenoceptors in CSV was elucidated. Contractions of tissue rings to the supramaximal concentrations of three selective alpha-2 agonists, B-HT 920, M-7 and clonidine, were determined in the absence and presence of 5 mM La . In the presence of La , clonidine and M-7 produced small but statistically significant contractions (8-14% of control) which were abolished when the alpha-1 adrenoceptors were inactivated by phenoxybenzamine (10(-7) M, 30 min). In contrast, contractions to B-HT 920 were abolished completely in the presence of La . All the three alpha-2 agonists stimulated 45Ca++ uptake into CSV (0.3-0.4 mmol/kg wet weight, 10 min). 45Ca++ efflux studies demonstrated that the selective alpha-2 agonist, B-HT 920 (10(-5) M plus 10(-7) M phenoxybenzamine), did not induce an increase in the rate of 45Ca++ efflux. In contrast, an augmented 45Ca++ efflux rate was observed with the alpha-1 agonist, phenylephrine (10(-4) M plus 10(-7) M rauwolscine). These results suggest that activation of postsynaptic alpha-2 adrenoceptors in CSV utilizes primarily extracellular Ca++ to produce contractions. PMID- 2989503 TI - Inhibition by metoprolol of the antihypertensive effect of aspirin in young rats. AB - A group of 21-day-old, spontaneous hypertensive rats (SHR) received either aspirin (75-100 mg/kg), metoprolol (1.0-1.5 mg/kg) or both in their drinking water for 56 days. Controls received plain water. Groups of normotensive Wistar Kyoto rats (WKY) received the same drug treatment as did the SHR. Blood pressure, heart rate and body weight were determined weekly, and water consumption was monitored on a daily basis. Renal prostaglandin (PG) activities were determined in a separate group of rats at 7 and at 56 days on each protocol. Blood pressures in the SHRs remained in the range of the WKY during 56 days of treatment with aspirin or metoprolol, but increased to hypertensive levels after exposure to plain water. Blood pressures in the WKY receiving aspirin or metoprolol were also lower than those receiving plain water, but the antihypertensive effect was somewhat slower in onset. The antihypertensive effects of aspirin and of metoprolol were lost in the SHR but not in the WKY when both drugs were given simultaneously in the drinking water. In comparison to rats receiving plain water, renal PG activity (PGF1 alpha and PGF2 alpha) was lower in rats receiving aspirin, metoprolol or both together. The mechanism for the antihypertensive effect of aspirin in the young SHR may be related to a decline in renal PG content, or to the release of other hormones controlling water and electrolyte balance. Reports that the cyclooxygenase inhibitors, such as aspirin, can block the antihypertensive effect of the beta adrenoceptor antagonists is confirmed in chronic studies using the SHR model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989504 TI - Serotonin-stimulated phosphoinositide turnover: mediation by the S2 binding site in rat cerebral cortex but not in subcortical regions. AB - In rat cerebral cortex, serotonin (5-HT) stimulates phosphoinositide turnover with an EC50 of 1 microM in the presence of pargyline. The EC50 is 16-fold higher in the absence of pargyline. Selective S2 antagonists inhibit 5-HT-stimulated phosphoinositide turnover. Schild analysis of the blockade by ketanserin of the 5 HT effect gives an estimated Kd of ketanserin for the phosphoinositide-linked receptor of 11.7 nM, which agrees with the Kd (3.5 nM) of [3H]ketanserin for the S2 site. Furthermore, MK-212, 5-HT and 5-fluorotryptamine stimulate phosphoinositide turnover with potencies that resemble their potencies at the S2 but not the S1 binding site. Of 11 agonists tested, the tryptamine derivatives tend to be more efficacious than the piperazine derivatives. The selective S1 agonist 8-hydroxy-2-(di-N-propylamino)tetralin is inactive at stimulating phosphoinositide turnover. No significant relationship exists between the regional distributions of 5-HT-stimulated phosphoinositide turnover and S2 binding sites. Furthermore, the S2 antagonist ketanserin is less potent and less efficacious in hippocampus and limbic forebrain than in cerebral cortex. These data suggest that 5-HT-stimulated phosphoinositide turnover is linked to the S2 binding site in rat cerebral cortex. However, 5-HT increases phosphoinositide turnover in subcortical regions by mechanisms other than stimulation of the S2 receptor. PMID- 2989505 TI - Release of leukotrienes, induced by the Ca++ ionophore A23187, from human lung parenchyma in vitro. AB - When chopped human lung is stimulated with the Ca++ ionophore A23187 (0.25-10 microM) leukotriene (LT) B4, LTD4 and LTE4 are found in the incubation medium according to different patterns. LTD4 is released promptly and its levels increase up to 45 min after the stimulus (A23187, 10 microM) and decline later (120 min); LTE4 formation follows a sigmoidal shape and continues to accumulate even 2 hr after the challenge; LTB4 levels reach a plateau at 45 min. LTC4 was undetectable in most experiments but it was found to accumulate when reduced glutathione (10 mM) was added. Addition of exogenous LTC4 to unstimulated fragments of human lung shows that an effective interconversion to LTD4 and LTE4 takes place: A23187 stimulates formation of LT and cyclo-oxygenase products dose dependently; a statistically significant formation of LT occurs at A23187 concentration of 1 microM whereas thromboxane B2 (TXB2) and 6-K-prostaglandin F1 alpha levels increased significantly only at 2.5 microM A23187. Pretreatment with U-60257 (100 microM) prevented formation of LT without a concomitant increase of TXB2 levels. Indomethacin (1.5 microM) blocked the release of 6-K-prostaglandin F1 alpha and TXB2 without a shift of arachidonic acid towards LT-like activity. Addition of exogenous LTC4 did not trigger synthesis of TXB2 or 6-K-prostaglandin F1 alpha. Our results indicate that reduced glutathione levels and the activity of the enzymes involved in LT biosynthesis and/or metabolism play an important role in controlling the pattern of LT release from human lung. PMID- 2989506 TI - Biologic activity of 3beta-D-glucopyranosides of vitamin D compounds. AB - Polar glycosidic conjugates of vitamin D compounds occur in the vegetable and possibly in the animal kingdom. The biologic activity of these conjugates has not been examined systematically. To obtain more information on the biological role of such sterol conjugates, we examined the biological activity of the 3beta-D glucopyranosyl conjugates of vitamin D3 5, 25-hydroxyvitamin D3 6, 1alpha hydroxyvitamin D3 7 and 1alpha,25-dihydroxyvitamin D3 8. When these compounds were administered i.v. we found that a dose of between 50 and 500 pmol/rat of the four glucopyranosides tested increased active intestinal calcium transport and increased bone calcium mobilization in vitamin D-deficient rats fed a low calcium diet. Under the same conditions, corresponding doses of the parent vitamin D3 compounds elicited comparable increases in both intestinal calcium transport and bone calcium mobilization. When these compounds were administered p.o. 3beta-D glucopyranosyl vitamin D3 5 exhibited no biological activity at doses of up to 5000 pmole/rat, whereas the corresponding glycosides of 25-hydroxyvitamin D3 6, 1alpha-hydroxyvitamin D3 7 and 1,25-dihydroxyvitamin D3 8 were active at doses of 500 to 1000 pmol/rat in the intestinal calcium transport system. When the glucopyranosyl conjugates were administered i.v. to vitamin D-deficient rats, 25 hydroxyvitamin D3 and 1alpha,-25-dihydroxyvitamin D3 were detected in the serum at levels less than or equal to those noted in animals dosed with the respective free sterols. PMID- 2989508 TI - Supraspinal convulsions induced by inverse benzodiazepine agonists in rabbits. AB - The electroencephalographic (EEG) effects of inverse benzodiazepine (BDZ) agonists have been studied in rabbits after i.v. administration. A dose-dependent progression of three different stages of EEG changes have been observed with inverse BDZ agonists. At first, trains of slow waves in the occipital cortex occur, followed by trains of spike-and-wave complexes in the sensorimotor cortex. These two stages are superimposed on a desynchronized cortical activity, accompanied by an enhancement of the hippocampal theta rhythm. These EEG changes parallel a state of alertness. The third stage is characterized by generalized grand-mal seizures made up of high voltage spikes in the cortical and subcortical brain areas accompanied by generalized tonico-clonic convulsions. No modification of electrical activity is observed at the level of the spinal cord. Methyl-beta carboline-3-carboxylate (beta-CCM) (at doses higher than 0.2 mg/kg) and 6,7 dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM) (at doses higher than 0.4 mg/kg) elicit all three stages, whereas ethyl-beta-carboline-3-carboxylate (beta CCE) (0.2-2 mg/kg) and N-methyl-beta-carboline-3-carboxamide (2-20 mg/kg) only elicit the first two, and finally CGS 8216 only the first. The extent of the EEG progression by inverse BDZ agonists may therefore be used as an index of the efficacy of each compound. The BDZ antagonists Ro 15-1788 and Ro 15-3505 (0.3 mg/kg or higher), which do not change the EEG pattern, block the effects of the convulsant and subconvulsant doses of the inverse BDZ agonists, giving rise to a desynchronized EEG pattern.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989507 TI - Differential effects of sulmazole (AR-L 115 BS) on contractile force and cyclic AMP levels in canine ventricular muscle: comparison with MDL 17,043. AB - MDL 17,043 [1,3-dihydro-4-methyl-5-[4-(methylthio)-benzoyl]-2H-imidazol-2-one] and AR-L 115 BS [sulmazole, 2-[(2-methoxy-4-methylsulfinyl)phenyl]-1H-imidazo[4,5 b] pyridine] produced substantial positive inotropic effects in a concentration dependent manner (10(-5) to 10(-3) M) in the isolated canine ventricular trabeculae in the presence of pindolol (3 X 10(-8) M). Both agents exhibited equal potency and intrinsic activity in this preparation. The time to peak tension, relaxation time and total duration of contraction were shortened by MDL 17,043 in a concentration-dependent manner up to the highest concentrations tested (10(-3) M). In contrast, AR-L 115 BS shortened these times in concentrations of 3 X 10(-4) M and lower, but prolonged them in higher concentrations. The positive inotropic effects of MDL 17,043 and AR-L 115 BS were associated with increases in the cyclic AMP levels of the muscles. The time course of the increase in cyclic AMP levels differed for the two drugs. The elevation in cyclic AMP and the positive inotropic effect produced by MDL 17,043 followed a similar time course. By contrast, the rise in the cyclic AMP level became significant only after the positive inotropic effect produced by AR-L 115 BS had reached a steady level. Carbachol, a muscarinic agonist, converted the positive inotropic effect of 10(-3) M MDL 17,043 into a negative inotropic effect, but did not alter the positive inotropic effect of 10(-3) M AR-L 115 BS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989509 TI - Evidence for gamma-aminobutyric acid mediation of the sympathetic nerve inhibitory response to vagal afferent stimulation. AB - The involvement of gamma-aminobutyric acid (GABA) in the vagal-stimulated reflex inhibition of sympathetic nerve discharge (SND) was investigated in the cat. Computer summation was used to assess the sympathoinhibitory response to vagal afferent stimulation and the resultant changes seen with agents known to effect GABAergic neurotransmission. The GABA antagonists picrotoxin and bicuculline at 0.25 to 1.0 mg/kg i.v. attenuated the vagal sympathetic inhibitory response, but increased the 1:1 locking of SND to the arterial pulse. Conversely, the GABA agonist diazepam at 0.3 mg/kg i.v. potentiated the vagal sympathoinhibition, reduced total SND and diminished SND locking to the arterial pulse. Picrotoxin also blocked the vagal sympathoinhibitory response in midcollicular transected cats and when applied topically to the dorsal brain stem. Midcollicular transection forestalled the irregular SND spiking normally seen with picrotoxin in the intact cat, thus resulting in improved 3 cycle/sec SND periodicity. None of these drug-induced changes effected the SND shutoff response to vasopressor tests, however. These results suggest that GABA plays a role in the SND inhibitory response to vagal afferent stimulation in the brain stem, independent of the sinoarterial baroreceptors, and may also be involved in the entrainment of SND to the arterial pulse. PMID- 2989510 TI - Effects of clenbuterol and antidepressant drugs on beta adrenergic receptor/N protein coupling in the cerebral cortex of the rat. AB - Repeated administration of the centrally acting beta adrenergic agonist clenbuterol to rats reduced the ability of isoproterenol to increase levels of cyclic AMP in slices of cerebral cortex. This lessened response to isoproterenol was not due to a reduction in beta receptor density but appeared to be due to diminished receptor/N-protein coupling. This was determined by measuring the ability of isoproterenol to inhibit the binding of the beta adrenergic antagonist [125I]iodopindolol to membranes prepared from cerebral cortex. Using membranes prepared from vehicle-treated rats, isoproterenol, in the absence of GTP, inhibited the binding of [125I]iodopindolol with an IC50 value of 85 nM and a Hill coefficient of 0.65. GTP (250 microM) increased the IC50 value to 290 nM and the Hill coefficient to 0.98. After repeated administration of 10mg/kg of clenbuterol to rats for 8 days, isoproterenol inhibited the binding of [125I]iodopindolol with an IC50 value of 125 nM and a Hill coefficient of 0.90; GTP increased the IC50 value to 170 nM and the Hill coefficient to 0.98. It was inferred from the results of modeling of the isoproterenol competition curves that the repeated administration of clenbuterol reduced or eliminated the high affinity component of isoproterenol binding. These effects of clenbuterol were found to depend on dose and duration of treatment and were reversible. Repeated administration of the antidepressant drugs desipramine, imipramine, phenelzine, zimelidine and mianserin twice daily for 21 days, by contrast, did not affect receptor/N-protein coupling.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989511 TI - Infusion of opiates into substantia nigra protects against maximal electroshock seizures in rats. AB - Microinfusion of morphine sulfate (50 nmol), [d-Ala2]-Met-enkephalin (35 nmol) or dynorphin A 1-13 (1 nmol) bilaterally into the substantia nigra significantly attenuated seizures induced by maximal electroshock in rats. This action was accompanied by stereotyped behavioral hyperactivity. These anticonvulsant and behavioral effects were antagonized by systemic naloxone administration; neither effect was observed after intranigral microinjection of dynorphin A 1-17 amide (1 nmol). These results are consistent with a mu opiate receptor-mediated inhibition of substantia nigra efferent neurons, and with the proposal that bilateral inhibition of nigral efferents attenuates seizure propagation. However, intranigral morphine failed to alter the severity of i.v. bicuculline seizures, indicating that opiate-mediated inhibition in substantia nigra is distinct from that produced by gamma-aminobutyric acid. PMID- 2989512 TI - [Percutaneous injection of a contrast medium in isolated lesions of the breast]. AB - The percutaneous injection of contrast medium was performed into 44 breast fibroadenomas, 13 tumors of fibrocystic disease and 5 cancers. The radiological aspect does not to assure some discrimination between peri- or intracanalicular fibroadenomas, nor between giant fibroadenomas and cystosarcoma phyllodes. The opacification of the intratumoral galactophoric channels is achieved in 34,1% of fibroadenomas. It can be observed in fibrocystic disease but nor in cancer. A pattern of fibrous tissue and/or opacification of large peripheric extratumoral channels will appear in 34,1% of fibroadenomas. They are be observed in fibrocystic diseases and cancers. It is the same thing for spot figure. For diagnosis, the technic is not recommended to be generalized since the accuracy seems to be less serious than the cytological analysis of the puncture. PMID- 2989513 TI - [Malignant fibrous histiocytoma of the mesentery. Echographic and x-ray computed tomographic aspects. Apropos of a case]. AB - We report one case of mesenteric malignant fibrous histiocytoma, demonstrated by pain, abdominal mass and hypoglycaemic faintness. Ultrasound showed an heterogeneous mass, with liquid areas surrounded with echogenic trabeculations. A percutaneous aspiration under ultrasound guidance of the liquid collections demonstrated sero-sanguineous material. A computed tomography showed the mass to involve the upper small bowel mesentery; organic hypoglycaemic series suggested a mesenchymal tumor, the histologic type of which was only known by post-mortem examination of the specimen. PMID- 2989514 TI - Relationship between cervical condylomata, pregnancy and subclinical papillomavirus infection. AB - From a retrospective study of 415 sets of colpophotographs, 25 clinically obvious condylomata of the cervix were identified. Two morphologically distinct forms were found and termed erythrocondylomata and leukocondylomata on the basis of their colposcopic appearance. Erythrocondylomata are red, raised lesions with diagnostic large capillary loops. Those lesions were found in young women (mean age, 22 years) and associated with pregnancy in 72% of cases. They were observed to involute and disappear to the naked eye within a few months of the diagnosis. Colposcopic and histologic studies, however, indicated that the lesions could persist in subclinical form. Of eight such lesions studied, five showed histologic and colposcopic features of subclinical papillomavirus infection, and two showed histologic features compatible with cervical intraepithelial neoplasia (CIN). Leukocondylomata are brilliant white lesions associated histologically with a thick layer of keratin over a typical condylomatous base. Those lesions occurred at a later age (mean, 32 years), were not associated with pregnancy and tended to increase rather than involute with time. Epidemiologic studies have indicated that it is the young and sexually active who are at most risk of developing cervical cancer. This study indicated that it is this same group that is also at highest risk of developing erythrocondylomata of the cervix. Such lesions have been shown to involve the squamocolumnar junction and transformation zone when that area is most active and vulnerable. The progression of such lesions to subclinical papillomavirus infection and CIN suggests that they may be involved in the initiation of the cervical neoplastic process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989515 TI - Human fibroblast interferon in cervical and vulvar intraepithelial neoplasia associated with viral cytopathic effects. A pilot study. AB - Sixteen patients with cervical intraepithelial neoplasia (CIN), grades 2 and 3, and vulvar intraepithelial neoplasia (VIN), grade 3, associated with human papillomavirus (HPV) infection were treated with human fibroblast interferon (HFI). Treatment consisted of 2-3 X 10(6) IU/day injected intra- and perilesionally for five days per week for two to three weeks with or without topically applied cream (1.2 X 10(6) IU/day for five days). Eight complete and two partial regressions occurred. The duration of the response ranged from 4 to 20 months. Fever, chills and fatigue with or without headache were the side effects. Our preliminary results indicate that HFI was active against CIN and VIN associated with HPV infection. PMID- 2989516 TI - Low-dose guar improves diabetic control. AB - Twenty diabetic outpatients (12 non-insulin-treated and 8 insulin-treated) were given guar granulate in a dose of 10 g daily for two months in order to study the effect on glycaemic control and lipid levels. Mean glycosylated haemoglobin levels (HbA1c%) fell from 11.1 +/- 2.0% pre-guar to 10.5 +/- 2.2% (P less than 0.001) after one month on guar and to 10.1 +/- 2.3% (P less than 0.0001) after two months. Following discontinuation of guar, HbA1c% rose to 11.1 +/- 2.5% (P less than 0.002). However, there were no significant changes in fasting blood glucose, 1 h postprandial blood glucose following a test meal, 24 h urinary glucose excretion or in lipid levels. Gastrointestinal side effects occurred in 4 patients during treatment with guar. Four patients reduced their dose of insulin and 2 patients reduced their dose of sulphonylurea therapy during this time because of symptoms suggestive of hypoglycaemia. We suggest that the low dose of guar used in this study may help improve glycaemic control in diabetic patients and that this may be achieved with a low incidence of gastrointestinal side effects. PMID- 2989517 TI - Isoxazoles with antipicornavirus activity. AB - The synthesis and evaluation of a series of 3,5-disubstituted isoxazoles as antipicornavirus agents have led to the discovery of several compounds effective in vitro against rhinovirus type 2 and poliovirus type 2. Compound 32 was found more effective than 4',6-dichloroflavan against both viruses and was evaluated orally in mice infected intracerebrally with polio-2. At 31 mg/kg bid, compound 32 showed a 53% survival rate as compared to 22% for the nonmedicated animals. PMID- 2989518 TI - Hashish: synthesis and central nervous system activity of some novel analogues of cannabidiol and oxepin derivatives of delta 9-tetrahydrocannabinol. AB - Several C-10 substituted cannabidiol (CBD) derivatives and novel oxepin derivatives of delta 9-tetrahydrocannabinol (delta 9-THC) were synthesized and evaluated for biological activity in mice and dogs. Treatment of 10 bromocannabidiol diacetate (3) with various amines in Me2SO gave the corresponding 10-aminocannabidiol derivatives 4-6. Similarly, treatment of 3 with NaN3 gave the azido compound 7, which with LiA1H4 afforded the 10 aminocannabidiol 9. However, reduction of 7 with CrCl2 formed the amide 8, which on further reduction with LiA1H4 gave the N-ethyl analogue 10. Coupling of 9 with 11 in the presence of dicyclohexylcarbodiimide formed 12, which was then deprotected with HCl to give the analogue 13. The oxepin analogue 14a was synthesized from 3 by treatment with Na2CO3 in CH3OH/H2O at room temperature. The dimethylheptyl analogue 14b was similarly prepared. Incorporation of N-ethyl (10), N-methyl-N-propargyl (6), and morpholino (4) groups in CBD at position 10 resulted in analogues that were more potent than CBD in producing hypoactivity in mice. These analogues had relatively little effect on rectal temperature. Selected substitutions at C-10 also resulted in analogues that were partially effective in blocking delta 9-THC antinociceptive activity. This blockade was observed particularly in compound 10, which also showed unusually toxic properties. Incorporation of a seven-membered oxepin in the delta 9-THC structure eliminated cannabinoid activity although substitution of the pentyl side chain with a 1,2-dimethylheptyl in the oxepin 14b resulted in CNS depression in mice. PMID- 2989519 TI - Synthesis and antiviral activity of 11-azapentacyclo[6.2.1.0.0.0]decane. AB - 11-Azapentacyclo[6.2.1.0.0.0]decane (6a) as well as its 6,7-dimethyl derivative 6b was synthesized by a novel, four-step sequence that holds promise for the construction of a variety of cage compounds with bridging nitrogen atoms. The hydrochloride salt of 6a was shown to possess no antiviral activity against either the influenza virus A/Victoria/3/75 or the herpes simplex viruses HSV-1 and HSV-2. PMID- 2989520 TI - 3-Amino-beta-carboline derivatives and the benzodiazepine receptor. Synthesis of a selective antagonist of the sedative action of diazepam. AB - Seven 3-N-substituted derivatives of 3-amino-beta-carboline were synthesized and their affinities for the benzodiazepine receptor were assessed in vitro. Two compounds, 3-(ethylamino)-beta-carboline and 3-[(methoxycarbonyl)amino]-beta carboline (beta-CMC), showing IC50 values of 460 and 71 nM, respectively, were selected for in vivo studies. The former compound showed long-lasting proconvulsant activity in Papio papio baboons while beta-CMC was shown in mice to selectively antagonize the sedative effects of diazepam without exhibiting convulsant, proconvulsant, or anxiogenic activity by itself. PMID- 2989521 TI - Synthesis and antiviral evaluation of nucleosides of 5-methylimidazole-4 carboxamide. AB - Due to the antiviral activity of certain 5-substituted imidazole nucleosides related to ribavirin, 5-methylimidazole-4-carboxamide nucleosides having beta-D ribofuranosyl, 2-deoxy-beta- and -alpha-D-ribofuranosyl, and (2 hydroxyethoxy)methyl moieties have been prepared and tested as antiviral agents. 1-beta-D-Ribofuranosyl-5-methylimidazole-4-carboxamide was obtained by deacetylation of the corresponding tri-O-acetyl nucleoside 11 or by deacetylation and ammonolysis of the blocked ethyl 5-methylimidazole-4-carboxylate nucleoside 10, which was prepared from the stannic chloride catalyzed condensation of the trimethylsilyl derivative of ethyl 4(5)-methylimidazole-5(4)-carboxylate. Glycosylation of 4(5)-methylimidazole-5(4)-carboxamide with 3,5-di-O-p-toluoyl-2 deoxy-D-erythro-pentofuranosyl chloride via mercuric cyanide method provided an anomeric mixture of the blocked 5-methylimidazole-4-carboxamide deoxynucleoside 14 along with an anomeric mixture of the 4-methyl 5-carboxamide isomer 15. Separation of compound 14 into the corresponding beta and alpha anomers was achieved by conversion to the 3',5'-di-O-acetyl derivatives 17 and 18, which after chromatographic separation were deacetylated to give 1-(2-deoxy-beta-D erythro-pentofuranosyl)-5-methylimidazole-4-carboxa mid e and its alpha anomer 20. 1-[(2-Hydroxyethoxy)methyl]-5-methylimidazole-4-carboxamide was prepared by alkylation of the imidazole 13 with (2-acetoxyethoxy)methyl bromide followed by treatment with methanolic ammonia. All these imidazole nucleosides were tested in HeLa cell cultures against type 1 herpes simplex and vesicular stomatitis viruses. The ribofuranosyl derivative 12 showed a significant activity against type 1 herpes simplex virus. PMID- 2989522 TI - Synthesis and biological activities of 2-pyrimidinone nucleosides. 2. 5-Halo-2 pyrimidinone 2'-deoxyribonucleosides. AB - 1-(2-Deoxy-beta-D-ribofuranosyl)-5-bromo-2-pyrimidinone (BrPdR) and 1-(2-deoxy beta-D-ribofuranosyl)-5-iodo-2-pyrimidinone (IPdR) have been synthesized by condensation of the appropriate silylated bases 2a and 2b, respectively, with 3,5 bis-O-(p-chlorobenzoyl)-2-deoxy-alpha-D-ribofuranosyl chloride (8) in 1,2 dichloroethane, in the presence of SnCl4, followed by separation of the anomeric blocked nucleosides via column chromatography and subsequent deprotection with methanolic ammonia. Both BrPdR and IPdR exhibited significant antiherpes activities against various strains of HSV-1 and HSV-2, the latter compound (IPdR) showing the higher activity as well as the stronger binding to the virus-specific thymidine kinase. PMID- 2989523 TI - Synthesis and antiherpetic activity of (S)-, (R)-, and (+/-)-9-[(2,3-dihydroxy-1 propoxy)methyl]guanine, linear isomers of 2'-nor-2'-deoxyguanosine. AB - Racemic 9-[(2,3-dihydroxy-1-propoxy)methyl]guanine [(+/-)-iNDG], a new analogue of acyclovir (ACV) and a structural analogue of 2'-nor-2'-deoxyguanosine (2'NDG), was synthesized and found to inhibit the replication of herpes simplex virus types 1 (HSV-1) and 2 (HSV-2). Subsequently, its optical isomers, (R)- and (S) iNDG, were prepared from chiral intermediates. The chloromethyl ethers of 1,2-di O-benzyl-D- and -L-glycerol were made and reacted with tris(trimethylsilyl)guanine to give the 9-alkylated guanines, which were deprotected by catalytic hydrogenolysis. Against HSV-1 and HSV-2 in cell culture, (S)-iNDG was approximately 10- to 25-fold more active than the R enantiomer and had an ED50 comparable to those for ACV and 2'NDG. The inferior activity of (R) iNDG paralleled the poor inhibition of viral DNA polymerase by its phosphorylation products. In mice infected intraperitoneally or orofacially with HSV-1 or intravaginally with HSV-2, (S)-9-[(2,3-dihydroxy-1 propoxy)methyl]guanine [(S)-iNDG] was less efficacious than 2'NDG but comparable to or more active than ACV. PMID- 2989524 TI - Synthesis, in vitro [3H]prazosin displacement, and in vivo activity of 3-aryl 4,5,6,7-tetrahydropyrazolo[4,3-c]pyridines, a new class of antihypertensive agents. AB - A series of new 3-aryl-4,5,6,7-tetrahydropyrazolo[4,3-c]pyridines was synthesized and screened for in vitro [3H]prazosin displacement activity. The results correlated well with their antihypertensive activity in spontaneous hypertensive rats. 1-Benzyl-3-(4-fluorophenyl)-4,5,6,7-tetrahydropyrazolo[4,3-c]pyrid ine (50, L 16052) was selected for further pharmacological evaluations of its potency when administered orally to conscious renal hypertensive dogs. PMID- 2989526 TI - Exclusion of the alpha 1(II) cartilage collagen gene as the mutant locus in type IA osteogenesis imperfecta. AB - Using two restriction site polymorphisms within the structural gene coding for human type II collagen we have examined the segregation of this gene in three pedigrees with dominantly inherited osteogenesis imperfecta (Sillence type IA). We have demonstrated that the gene does not segregate with clinical expression of the disease and cannot, therefore, contain the mutation responsible for osteogenesis imperfecta in these families. PMID- 2989525 TI - Linkage analysis of a DNA polymorphism proximal to the Duchenne and Becker muscular dystrophy loci on the short arm of the X chromosome. AB - The inheritance of a restriction fragment length polymorphism (RFLP) detected by a cloned DNA sequence (p754) from the short arm of the X chromosome has been studied in 14 Duchenne muscular dystrophy kindreds and six Becker muscular dystrophy kindreds. The linkage data obtained suggest that both the DMD and BMD loci are located in the same region (p21) on the short arm of the X chromosome at a distance of about 15 to 20 cM from the 754 locus. Data from families informative for both the p754 and L1.28 polymorphisms suggest that p754 is closer to the disease loci than is L1.28. PMID- 2989527 TI - Outer surface changes of Pseudomonas aeruginosa in relation to resistance to gentamicin and carbenicillin. AB - The outer surface structure of clinical isolates of Pseudomonas aeruginosa resistant to carbenicillin or gentamicin or both was studied by thin-section electronmicroscopy and compared with that of sensitive isolates. The latter had a fairly smooth outer layer. Strains resistant to both antibiotics were characterised by extrusions that appeared to constitute extensions of the outer membrane. The outer membrane appeared wavy with distortion of its tripartite structure. The latter findings were also present in isolates resistant to only one of the two antibiotics. The disorganisation of the outer membrane might contribute to the expression of resistance. PMID- 2989528 TI - Sodium-dependent modulation of the renal Na-K-ATPase: influence of mineralocorticoids on the cortical collecting duct. AB - Mineralocorticoids play a major role in the regulation of sodium transport in a variety of tissues, including the cortical collecting duct (CCD) of the mammalian nephron. To assess, in part, the underlying mechanism(s) of this control, the present studies were designed to evaluate, first, the influence of mineralocorticoids on the Na-K-ATPase activity in the rabbit CCD and, secondly, a possible role of sodium entry into the cell at the luminal border on the regulation of the Na-K-ATPase. In the first series of studies, rabbits were maintained on a low sodium diet which raised serum aldosterone levels from 16 to 70 ng/dl after 3-4 days, with further elevations being expressed with treatment for two weeks or more. In CCDs isolated from these animals, the Na-K-ATPase increased from 13 to 40 pmol ADP min-1 mm-1 after 3-4 days on the low sodium regimen, but then declined, returning to control values after approximately 2 weeks. This decline in activity was preceded by a decrease in the Na+ concentration of the urine to low levels and hence, likely coincided with a decreased delivery of sodium to, and sodium entry into the cells of, the CCD. If dietary manipulations were used to maintain a high delivery of sodium to the CCD in the animal, elevation of plasma mineralocorticoid levels by treatment with deoxycorticosterone acetate (DOCA) caused a similar elevation in the Na-K-ATPase activity after 3-4 days, which did not decline with continued treatment for up to 2 weeks. Furthermore, it was observed that mineralocorticoids only exerted their effect on the Na-K-ATPase after a latent period of 1 day, well after sodium excretion had fallen, indicating that sodium entry into the CCD cells was already stimulated. If animals were simultaneously treated with DOCA and the sodium channel blocker amiloride for 3-4 days, the effects on the Na-K-ATPase were markedly reduced, whereas amiloride treatment alone had no effect on the enzyme activity. Since others have shown that mineralocorticoids induce synthesis of the Na-K-ATPase subunits in toad bladder cells in an amiloride-insensitive manner, sodium must be exerting its effect on a process after translation. It is concluded that the initial effect of mineralocorticoids in the CCD is on sodium entry with a delayed induction of the Na-K-ATPase, which is regulated by Na dependent modulation of a posttranslational process. PMID- 2989529 TI - Mouse alpha-amylase loci, Amy-1a and Amy-2a, are closely linked. AB - We have cloned a contiguous 106 X 10(3) base-pair long stretch of mouse DNA. The isolated chromosomal DNA segment contains the single copy gene Amy-1a that is strongly expressed in the parotid gland and, 23 X 10(3) base-pairs downstream from it, one member of the pancreas-specific Amy-2a oligogene family. At least two of the four Amy-2a genes, including the copy linked to Amy-1a, are efficiently transcribed. The cloned DNA sequences do not appear to specify messenger RNAs other than those encoding alpha-amylase in pancreas, parotid gland or liver. Transcription termination on Amy-1a occurs within 3 X 10(3) base-pairs downstream from the polyadenylation site in both parotid gland and liver, in which this gene is transcribed at different rates from different promoters. PMID- 2989530 TI - Characterization of the rat gamma-crystallin gene family and its expression in the eye lens. AB - Rat genomic clones, which together contain all of the rat genomic gamma crystallin sequences, have been characterized. Five gamma-crystallin genes are located on a contiguous DNA region, 63 X 10(3) base-pairs long. These genes, named (5') gamma 1-1, gamma 1-2, gamma 2-2 and gamma 3-1 (3'), are all oriented head to tail. A sixth gamma-crystallin gene, named the gamma 4-1 gene, could not be linked to the gamma-crystallin gene cluster with our present set of genomic clones. Mapping experiments using single copy sequences which form the extreme 5' or 3' region of the gene cluster showed that, if the gamma 4-1 gene is located on the same chromosome, then it must be separated from the gene cluster by at least 25 X 10(3) base-pairs of DNA. All gamma-crystallin genes have a similar mosaic structure. They contain a large (0.9 X 10(3) to 1.88 X 10(3) base-pairs) intron in the middle of the gene and are further interrupted close to the 5' end of the gene. The length of the first exon varies from about 40 to about 50 base-pairs. The complementary DNA clone pRL-gamma-3 used in this study is a copy of the transcript of the gamma 3-1 gene, while the second complementary DNA clone, pRL gamma-2, is most likely a copy of the transcript of the gamma 2-1 gene. It is further shown that rat lens messenger RNA protects fragments from the 3' ends of the four other gamma-crystallin genes against degradation by S1 nuclease, hence all six gamma-crystallin genes present in the rat genome must be transcribed in the lens. Repetitive sequences were found to be present between and around the gamma-crystallin genes. Mapping with cloned repetitive sequences showed that three different repeats, designated A, B and C, occur more than once in the gamma crystallin gene cluster. Repeat C is also found in the gamma 4-1 region. A repetitive region 3' to the gamma 3-1 gene contains members of all three repeat families. PMID- 2989531 TI - Sequences essential for IS50 transposition. The first base-pair. AB - Sequences near the ends of the insertion element IS50 are essential for its transposition, probably because they serve as sites upon which the IS50-encoded transposase protein acts. To determine if these essential sequences include the first base-pair at each end of IS50 we generated 5'C to 5'G transversions at these positions. Each mutation reduced the transposition frequency to 1% to 2% of wild-type. DNA sequence analyses showed that the mutant 5'G is preserved during transposition. PMID- 2989532 TI - Bacteriophage P4 DNA replication. Location of the P4 origin. AB - An electron microscopic examination of replicating bacteriophage P4 DNA molecules has revealed theta-type structures that replicate bidirectionally from a single origin. Many replicating P4 DNA molecules also contain long (2000 bases) single strand DNA regions at the growing fork that are deployed in a trans configuration, which supports the concept of continuous leading strand and discontinuous lagging strand syntheses. The position of the P4 origin was localized by the use of a plasmid complementation test for replication in vivo, as well as by labeling of DNA replicating in vitro in the presence of a chain terminating inhibitor. During this study we discovered a second site on the P4 genome which is essential for replication, and we have named it crr (cis region required for replication). The site is located at least 3300 bases from the origin but appears to be required for the initiation of DNA replication in vivo as well as in vitro. PMID- 2989533 TI - Messenger RNAs from the transforming region of bovine papilloma virus type I. AB - Messenger RNAs present in C127 mouse cells transformed by bovine papilloma virus type 1 (BPV-1) were studied by the S1 nuclease protection technique, Northern blotting, and electron microscopic heteroduplex analysis. The results revealed at least five classes of spliced mRNAs which we designate types 1 to 5. They had a common poly(A) addition site located at co-ordinate 53 and all mRNAs, except the type 3 mRNAs, contained an exon located between co-ordinates 41 and 53. In the type 1 mRNAs this exon was connected to a very short leader sequence located around co-ordinate 31. The type 2 mRNAs contained 220 to 400-nucleotide long leaders which were located approximately 1.5 X 10(3) base-pairs further upstream. Two different subclasses of type 2 molecules (2A and 2B) were identified and these had slightly different leaders. The type 4 mRNAs contained a bipartite leader, whereas the type 5 mRNAs carried an approximately 900-nucleotide long leader. The type 3 mRNAs consisted of a main exon located between co-ordinates 32 and 53, linked to the same leader as is present in the type 2A mRNAs. A cap site which presumably is utilized by the type 2A, type 3, type 4 and type 5 mRNAs was mapped at nucleotide 89 in the BPV-1 sequence. A putative cap site for the type 1 mRNAs was mapped at co-ordinate 31. PMID- 2989534 TI - Inhibition of the type I restriction-modification enzymes EcoB and EcoK by the gene 0.3 protein of bacteriophage T7. AB - The gene 0.3 protein of bacteriophage T7 is a potent inhibitor of the restriction modification enzymes EcoB and EcoK, both in vivo and in vitro. We have analyzed the ability of purified 0.3 protein to inhibit different steps in the reactions of EcoB and EcoK with DNA. Most of our experiments were done with EcoK, but selected tests with EcoB indicate that the two enzymes are affected by 0.3 protein in the same way. Purified 0.3 protein binds tightly to free enzyme, apparently to one of the small subunits, and prevents it from binding to DNA. If EcoK is allowed to form specific recognition complexes with unmodified DNA before 0.3 protein is added, relatively low levels of 0.3 protein prevent the nuclease activity that would otherwise appear upon addition of ATP, but considerably higher levels are needed to prevent formation of filter-binding complexes or ATPase activity. This, together with other results, suggests that the binding site for 0.3 protein is protected in recognition complexes and in the early stages of the ATP-stimulated reactions, but that it becomes accessible again before cleavage of the DNA, perhaps after the translocation step. If added after the nuclease reaction is substantially over, 0.3 protein has little effect on ATPase activity, and indeed, the subunit having the binding site for 0.3 protein apparently dissociates from the enzyme-DNA complex. The methylase activity of EcoK on hemi-methylated recognition sites is strongly inhibited by 0.3 protein added at any stage of the reaction. PMID- 2989535 TI - Two type I restriction enzymes from Salmonella species. Purification and DNA recognition sequences. AB - We have purified the type I restriction enzymes SB and SP from Salmonella typhimurium and S. potsdam, respectively, and determined the DNA sequences that they recognize. These sequences resemble those previously determined for the type I enzymes, EcoB, EcoK and EcoA, in that the specific part of the sequence is divided into two domains by a spacer of non-specific sequence that has a fixed length for each enzyme. Two main differences from the previously determined sequences are seen. Both of the new sequences are degenerate and one of them, SB, has one trinucleotide and one pentanucleotide-specific domain rather than the trinucleotide and tetranucleotide domains seen for all of the other enzymes. The only conserved features of the recognition sequences are the adenosyl residues that are methylated in the modification reaction. For all of the enzymes these are situated ten or 11 base-pairs apart, one on each strand of the DNA. This suggests that the enzymes bind to DNA along one face of the double helix making protein-DNA interaction in two successive major grooves with most of the non specific spacer sequence in the intervening minor groove. PMID- 2989536 TI - Apparent alteration in properties of arl mutants of Escherichia coli. AB - The published properties of lambda phages grown in Escherichia coli arl mutants, and plasmids maintained in them, included increased homologous recombination, decreased DNA-cytosine methylation, and increased sensitivity of DNA to nuclease S1. Some of these properties now appear altered; others remain approximately as published. PMID- 2989537 TI - Electrostatic interactions in globular proteins: calculation of the pH dependence of the redox potential of cytochrome c551. AB - An equal mixture of oxidized and reduced cytochrome c551 experiences a change in the potential of the haem iron when one of the propionates attached to the haem is ionized. The change is 65 mV, which corresponds to an effective dielectric between the propionate and the iron of 27. It has been possible to use the algorithm of Warwicker & Watson (1982) to calculate the change in the potential at the haem iron arising from the change in ionization of the propionate. This gives the extra work required to oxidize or reduce the iron. The change in potential that we calculate is 90 mV, which corresponds to an effective dielectric of 19.5, between the propionate and the iron. In comparison with other commonly used dielectric models the agreement is very good. PMID- 2989538 TI - Novobiocin blocks the Drosophila heat shock response. AB - In the studies reported here we show that the antibiotic novobiocin, an in vitro inhibitor of topoisomerase II, blocks the Drosophila heat shock response. If novobiocin is added prior to induction, there is no detectable expression of the Drosophila heat shock genes. Moreover, analysis of the chromatin organization of the 87A7 heat shock locus indicates that the antibiotic prevents the structural alterations which normally accompany heat induction. When novobiocin is added after induction, transcription appears to be rapidly turned off, and the chromatin organization of the 87A7 locus is "fixed" in an "active" configuration. Novobiocin also prevents the re-establishment of the pre-induced 87A7 chromatin organization which occurs during recovery from heat shock. We have also presented data suggesting that this antibiotic blocks transcription at 25 degrees C. These findings raise the possibility that topoisomerase II may be required in eukaryotes for both gene activation and deactivation. PMID- 2989539 TI - Partial suppression of an ochre mutation in Saccharomyces cerevisiae by multicopy plasmids containing a normal yeast tRNAGln gene. AB - We screened a yeast genomic library for recombinant DNA plasmids that complemented the ultraviolet (u.v.) sensitivity of a strain of Saccharomyces cerevisiae designated rad4-3 that is defective in excision repair of DNA. A multicopy plasmid (pNF4000) with a 9.4 X 10(3) base-pair yeast DNA insert partially complemented the u.v. sensitivity of rad4-3, but not of two other rad4 allelic mutants (rad4-2 and rad4-4), or of other u.v.-sensitive rad mutants. The yeast insert was analyzed by restriction mapping, DNA-DNA hybridization, DNA-tRNA hybridization and DNA sequencing. This analysis revealed the presence of a normal tRNAGln gene, a yeast sigma element situated 5' to the transfer RNA gene, a Ty element and a solo delta element. Deletion analysis of pNF4000 showed that the tRNAGln gene is required for partial complementation of the u.v. sensitivity of rad4-3. Furthermore, a multicopy plasmid containing a tRNAGln gene derived from a different region of the yeast genome also partially complemented the u.v. sensitivity of rad4-3. The rad4-3 mutation is suppressed following transformation with a plasmid containing the known ochre suppressor SUP11-o, indicating that it is an ochre mutation. We therefore conclude that when expressed in sufficient quantity, normal tRNAGln (which usually decodes the sense codon CAA) can weakly suppress the nonsense ochre codon UAA, and suggest that this represents an example of wobble occurring at the first rather than at the third position of the codon. PMID- 2989540 TI - Differential expression of two clusters of mouse histone genes. AB - The mouse histone mRNAs coded for by three different cloned DNA fragments have been characterized. Two of these cloned DNA fragments, MM221 and MM291, located on chromosome 13, code for H3, H2b and H2a histone mRNAs, which are expressed at low levels in cultured mouse cells and fetal mice. The other DNA fragment, MM614, located on chromosome 3, codes for an H3 and an H2a mRNA, which are expressed at high levels in these cells. The mRNAs for each histone protein share common coding region sequences, while the untranslated regions of all the genes have diverged significantly, as judged by S1 nuclease mapping. Amino acid substitutions in some H3, H2a and H2b proteins are detected as internal cleavages in the S1 nuclease maps. All of these genes code for replication variant histone mRNAs, which are regulated in parallel with DNA synthesis. PMID- 2989541 TI - Molecular analysis of the heterogeneity region of the human ribosomal spacer. AB - The human ribosomal non-transcribed spacers are 30 X 10(3) base-pairs (or 30 kb) in length with a limited length heterogeneity localized in a specific region downstream from the 3' end of the transcribed region. Total DNA digested with EcoRI and BamHI and hybridized with a probe containing the 3' end of the 28 S ribosomal RNA coding region shows four major bands of 3.9 kb, 4.6 kb, 5.4 kb and 6.2 kb. The 5.4 kb band is the most abundant in every individual, followed by the 4.6 kb band. The longest and the shortest size classes are less well-represented and may even be absent. Every individual shows his own pattern of relative abundance of non-transcribed spacer length classes that can be followed through generations. We decided to investigate the molecular structure of the heterogeneity region, in order to cast light onto the mechanisms underlying the origin and maintenance of this length heterogeneity. Pertinent spacer regions of eight ribosomal clones from two human genomic libraries were subcloned and analyzed by restriction mapping and nucleotide sequencing. In the minimal length class, there is a sequence of 700 base-pairs that appears to be tandemly duplicated once, twice or three times in the other length classes. This repeated DNA module contains a region consisting of repetitions of simple pyrimidine groups like C-T, C-T-T-T or C-C-C-T. DNA module repeats may differ by the length of this pyrimidine-rich region. However, these length variations are not continuous, as revealed by Southern transfer analysis of several individuals and different cloned gene units: instead, the repeated modules fall into two discrete length classes of about 700 base-pairs and 800 base-pairs. An imperfect duplication of a short sequence of 86/89 base-pairs is present at the boundary between the heterogeneity region and the upstream flanking region, representing a very ancient duplication event. PMID- 2989542 TI - The terminase of bacteriophage lambda. Functional domains for cosB binding and multimer assembly. AB - Terminase is a protein complex involved in lambda DNA packaging. The subunits of terminase, gpNul and gpA, are the products of genes Nul and A. The actions of terminase include DNA binding, prohead binding and DNA nicking. Phage 21 is a lambdoid phage that also makes a terminase, encoded by genes 1 and 2. The terminases of 21 and lambda are not interchangeable. This specificity involves two actions of terminase; DNA binding and prohead binding. In addition, the subunits of lambda terminase will not form functional multimers with the subunits of 21 terminase. lambda-21 hybrid phages can be produced as a result of recombination. We describe here lambda-21 hybrid phages that have hybrid terminase genes. The packaging specificities of the hybrids and the structure of their genes were compared in order to identify functional domains of terminase. The packaging specificities were determined in vivo by complementation tests and helper packaging experiments. Restriction enzyme site mapping and sequencing located the sites at which recombination occurred to produce the hybrid phages. lambda-21 hybrid 51 carries the lambda A gene, and a hybrid 1/Nul gene. The crossover that produced this phage occurred near the middle of the 1 and Nul genes. The amino-terminal portion of the hybrid protein is homologous to gp1 and the carboxy-terminal portion is homologous to gpNul. It binds to 21 DNA and forms functional multimers with gpA, providing evidence that the amino-terminal portion of gpNul is involved in DNA binding and the carboxy-terminal portion of gpNul is involved in the interaction with gpA. lambda-21 hybrid 54 has a hybrid 2/A gene. The amino terminus of the hybrid protein of lambda-21 hybrid 54 is homologous with gp2. This protein forms functional multimers only with gp1, providing evidence that the amino terminus of gpA is involved in the interaction with gpNul. These studies identify three functional domains of terminase. PMID- 2989543 TI - Essential interaction between lambdoid phage 21 terminase and the Escherichia coli integrative host factor. AB - Lambdoid phage 21 requires the Escherichia coli integrative host factor (IHF) for growth. lambda-21 hybrids that have 21 DNA packaging specificity also require IHF. IHF-independent (her) mutants have been isolated. her mutations map in the amino-terminal half of the 21 1 gene. The 1 gene encodes the small subunit of the 21 terminase, and the amino-terminal half of the 1 polypeptide is a functional domain for specifically binding 21 DNA. Hence changes in the DNA-binding domain of terminase, her mutations, render 21 terminase able to function in the absence of IHF. Three of four her mutations studied are trans-dominant. An in vitro system was used to show that packaging of 21 DNA is IHF-dependent. IHF is directly required during the early, terminase-dependent steps of assembly. It is concluded that IHF is a host factor required for function of the 21 terminase. It is proposed, in analogy to the role of IHF in lambda integration, that IHF facilitates proper binding of 21 terminase to phage DNA. Consistent with this proposal, possible IHF-binding sites are present in the 21 cohesive end site. PMID- 2989544 TI - Effects of perfusion pressure on the bursting neurons in the intact or segmented cardiac ganglion of the lobster, Panulirus japonicus. AB - Impulse activity of the bursting neurones in the intact or segmented cardiac ganglion could be recorded in the isolated and freely beating hearts of lobsters. In the segmented ganglion, the four large cells (LCs), the medium cell (MC), and the four small cells (SCs) generated phasic, short tonic, and long tonic bursts of impulses with different rhythms, respectively. The periodic bursts of the LCs, the MC, and the SCs in the intact and the segmented ganglia were perturbed by a change of heart tonus which reflected a change of perfusion pressure in the heart. When correlation between period and duration of the perturbed bursts was plotted, the LCs, the MC, and the SCs in the segments were different from one another in distribution of the plots. Further, the correlation plots for the bursts of the MC in the intact ganglia were compared to those of the LCs, the MC, and the SCs in the segmented ganglia. The burst period and duration of the MC in the intact ganglion were altered markedly by a change of the burst period of the LCs and the burst duration of the SCs, as well as by change of heart tonus. PMID- 2989545 TI - The spectrum of myocardial contusion: a review. AB - Blunt trauma to the chest and abdomen frequently results in cardiac injury. A wide spectrum of pathology can follow, including myocardial concussion and contusion, valvular disruption, and pericardial effusion and tamponade. Likewise, sequelae may be inconsequential or lead to sudden death, and may occur immediately at the time of trauma or be delayed by days to years. Despite the increasingly common occurrence of myocardial contusion there remains much confusion as to how the diagnosis is made. The various diagnostic studies utilized are frequently misinterpreted. The pathophysiology, clinical presentation, and a critical evaluation of the diagnostic tests used in the confirmation of this entity are reviewed, and an approach to the evaluation and management of these patients is presented. PMID- 2989546 TI - A rationale for periodontal therapy in the Keyes era. PMID- 2989547 TI - Intraoperative ultrasound guidance for needle biopsy of a contralateral intrarenal "tumorlet" in a child with Wilms' Tumor. PMID- 2989549 TI - Molecular relationships among serogroup B bacteriophages of Staphylococcus aureus. AB - The typing bacteriophages 55, 80, 83A, and 85 of Staphylococcus aureus, representative of the three major lytic groups of serological group B aureophages, have been examined for relatedness of their genomes and virion proteins. Phages 11 and 80 alpha were also examined to determine the relationship of phage 80 alpha to phages 11 and 80. Total genome hybridization measurements divided the phages into two groups. Phages 55 and 80, in the first group, had DNA homology of 50%. Phages 11, 80 alpha, 83A, and 85 formed a second group with 27 to 65% homology. Homology between the two groups was in the range of 14 to 22%. Phage 80 alpha is more closely related to phage 11 than to phage 80, though it is probably not a simple recombinant of phages 11 and 80. Restriction enzyme digestion and phage [32P]DNA hybridization analysis of the endonuclease-generated fragments from each phage DNA confirmed the findings of the DNA homology measurements. The endonuclease fragment patterns generated by EcoRI and HindIII were distinctive for each phage, confirming that none of the phages are closely related. Common sequences were present in most fragments from the phage DNAs when the labeled probe DNA was from a different phage in the same group. Cross-group probing of endonuclease fragments revealed both a diminished level of homology when similar sequences were present and the probable absence of some sequences. Virion proteins, examined by polyacrylamide gel electrophoresis, were similar in number and molecular weight for phages 11, 80 alpha, 83A, and 85, reflecting the DNA homology analyses. The virion proteins from phages 55 and 80, however, were more distinctive, and both differed from the phages in the other group. PMID- 2989548 TI - Site-directed mutagenesis of the simian virus 40 large T-antigen gene: replication-defective amino acid substitution mutants that retain the ability to induce morphological transformation. AB - We used a heteroduplex deletion loop mutagenesis procedure for directing sodium bisulfite-induced mutations to specific sites on viral or plasmid DNA to generate a series of SV40 large T-antigen point mutants. The mutations were directed to a region of the T-antigen gene, 0.5 map units, that is thought to be important for interaction of the protein with the viral origin of DNA replication. Of the 16 mutants reported here, 10 had lost the ability to replicate their DNA, and 3 others showed a reduced level of replication compared to wild type. All of the mutants tested were capable of transforming rat cells in culture by the dense focus assay. We conclude that the sequences of the early region around 0.5 map units are critical for the replication of viral DNA but not for the transformation function of T antigen. PMID- 2989550 TI - Terminal repetitive sequences in herpesvirus saimiri virion DNA. AB - The H-DNA repeat unit of Herpesvirus saimiri strain 11 was cloned in plasmid vector pAGO, and the nucleotide sequence was determined by the dideoxy chain termination method. One unit of repetitive DNA has 1,444 base pairs with 70.8% G+C content. The structural features of repeat DNA sequences at the termini of intact virion M-DNA (160 kilobases) and orientation of reiterated DNA were analyzed by radioactive end labeling of M-DNA, followed by cleavage of the end fragments with restriction endonucleases. The termini appeared to be blunt ended with a 5'-phosphate group, probably generated during encapsidation by cleavage in the immediate vicinity of the single ApaI recognition site in the H-DNA repeat unit. The sequence did not reveal sizeable open reading frames, the longest hypothetical peptide from H-DNA being 85 amino acids. There was no evidence for an mRNA promoter or terminator element, and H-DNA-specific transcription could not be found in productively infected cells. PMID- 2989551 TI - Nucleotide sequence and structural features of a novel US-a junction present in a defective herpes simplex virus genome. AB - Defective genomes generated during serial propagation of herpes simplex virus type 1 (Justin) consist of tandem reiterations of sequences that are colinear with a portion of the S component of the standard viral genome. We determined the structure of the novel US-a junction, at which the US sequences of one repeat unit join the a sequences of the adjacent repeat unit. Comparison of the nucleotide sequence at this junction with the nucleotide sequence of the corresponding US region of the standard virus genome indicated that the defective genome repeat unit arose by a single recombinational event between an L-S junction a sequence and the US region. The recombinational process might have been mediated by limited sequence homology. The sequences retained within the US a junction further define the signal for cleavage and packaging of viral DNA. PMID- 2989552 TI - Structure and physical map of Rhodopseudomonas sphaeroides bacteriophage RS1 DNA. AB - We analyzed, by restriction endonuclease mapping and electron microscopy, the genome of the lytic Rhodopseudomonas sphaeroides-specific bacteriophage RS1 and characterized it as a linear molecule of approximately 60 to 65 kilobases. When the DNA from purified phage particles was examined by several independent methods, considerable size heterogeneity was apparent in the RS1 DNA. This size heterogeneity was concluded to be of biological origin, was independent of the specific host strain used to propagate virus, and was not due to the presence of host DNA within or nonspecifically associated with purified virions. In addition, treatment of RS1 DNA with either BAL 31 nuclease or DNA polymerase I Klenow fragment revealed that several distinct regions exist within the viral chromosome which contain free 3' hydroxyl groups. A restriction endonuclease map of the RS1 genome was constructed by using the restriction endonucleases EcoRI, ClaI, KpnI, BamHI, MluI, SmaI, and BclI; thereby allowing the positioning of some 40 restriction sites within the viral genome. The results are discussed in terms of the significance and the possible biological origin of the unique features discovered within the phage RS1 DNA. PMID- 2989553 TI - Isolation of cis-acting vaccinia virus DNA fragments promoting the expression of herpes simplex virus thymidine kinase by recombinant viruses. AB - Recombinant TK- vaccinia viruses containing the pBR322 sequence inserted in either orientation within the coding sequence of the viral thymidine kinase gene were constructed. They were characterized by genomic analysis, hybridization studies, reversion to wild-type virus by in vivo recombination, and rescue from their genomes of plasmids which contained all or parts of the pBR322 sequence. TK cells were infected with one of these recombinant viruses and then transfected with pools of chimeric plasmids composed of a cloned herpes simplex virus thymidine kinase gene which contained upstream inserts of different vaccinia DNA fragments prepared by restriction or sonication. Recombination between homologous pBR322 sequences within infected cells generated selectable recombinant viruses in which expression of the herpes simplex virus thymidine kinase gene was promoted by the upstream vaccinia insert. These viruses were characterized by genomic analysis, hybridization, and in vivo or in vitro phosphorylation of (5 [125I]deoxycytidine as a specific assay for the expressed herpes simplex virus thymidine kinase. Vaccinia DNA inserts were isolated conveniently for transfer to bacteria by rescuing appropriate plasmids from the genome of recombinant viruses. The sequence of 100 nucleotides adjacent to the upstream region of the herpes simplex virus gene was determined in nine different inserts measuring 0.17 to 1.07 kilobase pairs. PMID- 2989554 TI - Nucleotide sequence of a transduced myc gene from a defective feline leukemia provirus. AB - The nucleotide sequence of a feline v-myc gene and feline leukemia virus (FeLV) flanking regions was determined. Both the nucleotide and predicted amino acid sequences are very similar to the murine and human c-myc genes (ca. 90% identity). The entire c-myc coding sequence is represented in feline v-myc and replaces portions of the gag and env genes and the entire pol gene. The coding sequence is in phase with the gag gene reading frame; v-myc, therefore, appears to be expressed as a gag-myc fusion protein. Viral sequences at the 3' myc-FeLV junction begin with the hexanucleotide CTCCTC, which is also found at the 3' fes FeLV junction of both Gardner-Arnstein and Snyder-Theilen feline sarcoma viruses. These similarities suggest that some sequence specificity may exist for the transduction of cellular genes by FeLV. Feline v-myc lacks a potential phosphorylation site at amino acid 343 in the putative DNA-binding domain, whereas both human and murine c-myc have such sites. Avian v-myc has lost a potential phosphorylation site which is present in avian c-myc five amino acids from the potential mammalian site. If these sites are actually phosphorylated in normal c-myc proteins, their loss may alter the DNA-binding affinity of v-myc proteins. PMID- 2989555 TI - Transformation of Brown Leghorn chicken embryo fibroblasts by avian myeloblastosis virus proviral DNA. AB - Brown Leghorn chicken embryo fibroblasts were transfected with a mixture of avian myeloblastosis virus (AMV) and myeloblastosis-associated virus type 1 (MAV1) proviral DNA purified from lambda-Charon 4A recombinant clones. A transformed cell line (T1AM) able to grow without anchorage in semisolid medium was obtained. The presence of both proviral AMV and MAV sequences was detected in T1AM DNA by hybridization with v-myb- and MAV1-specific probes. Altered AMV and MAV1 proviral genomes were found in T1AM genome. Characterization of the RNA species expressed in transformed cells showed that in addition to a 2.5-kilobase (kb) putative subgenomic v-myb-specific RNA, three other myb-containing RNAs (9.4, 8.4, and 7.0 kb) were present in T1AM cells. No AMV genomic RNA was detected. Also, a new 5.0 kb MAV1-specific RNA species was expressed in transformed cells in addition to MAV1 genomic RNA species (7.8 kb). No infectious AMV virions are released by T1AM cells. Chicken embryo fibroblasts infected by T1AM-released virions contained and expressed all MAV1 sequences detected in T1AM transformed cells but did not express any transformation parameter. These results indicated that the presence of AMV proviral sequences in T1AM cells is responsible for their transformed phenotype. PMID- 2989557 TI - Proviral genome of radiation leukemia virus: molecular cloning of biologically active proviral DNA and nucleotide sequence of its long terminal repeat. AB - The proviral genome of a leukemogenic and thymotropic C57BL/Ka mouse retrovirus, RadLV/VL3(T+L+), was cloned as a biologically active PstI insert in the bacterial plasmid pBR322. Its restriction map was compared with those, already known, of two nonthymotropic and nonleukemogenic viruses of the same mouse strain: the ecotropic BL/Ka(B) virus and the xenotropic constituent of the radiation leukemia virus complex. Differences were observed around the gag-pol gene junction, in the pol gene, and in the env gene. Moreover, the nucleotide sequence of the RadLV/VL3(T+L+) long terminal repeat revealed the existence of two copies of a 43 base-pair sequence, of which BL/Ka(B) possesses only one copy. PMID- 2989556 TI - Developmental and molecular aspects of nephroblastomas induced by avian myeloblastosis-associated virus 2-O. AB - Avian myeloblastosis-associated virus-induced nephroblastomas are tumors consisting mainly of mesenchymal and epithelial renal elements with variable degrees of differentiation. The spatial distribution of developmental stages reflects a gradient of differentiation from less differential structures in the periphery towards more differentiated structures in the center of the lobules formed in the nephroblastomas. These heterogenic tumors contain discrete virus cell DNA junction fragments and are therefore clonal outgrowths of a single transformed cell. These findings support the hypothesis that a mesenchymal, nephrogenic cell residual in the postembryonic kidney is the origin of the tumor, which grows by proliferation and differentiation of this target cell. All the tumors expressed higher levels of viral genomic and env messages than nontransformed tissue from the same kidney. A screening of oncogene expression with 13 different oncogenes revealed enhanced myc levels. There was, however, no rearrangement of c-myc or of the other oncogenes detected with EcoRI-digested tumor DNAs. This suggests that there is no insertion of viral elements adjacent to a c-myc. The levels of myc expression in embryonic kidneys were as high as in the tumors. Therefore, the enhanced myc expression in nephroblastomas is a reflection of the embryonic status of the tumor rather than a newly acquired function. This finding, plus the similarity of development and morphology of nephroblastomas and embryonic kidneys, suggests that the tumors arise as a result of a deficiency in a function which turns the embryonic status off. PMID- 2989558 TI - Deletion mutants of Bacillus subtilis bacteriophage SP beta. AB - The central portion of the chromosome of temperate Bacillus subtilis bacteriophage SPB was found to contain a region in which large deletions occurred, sometimes at high frequency. Most of the deletions could be placed into one of three groups, del1, del3, and del4, which were missing 11.8, 14.2, and 14 kilobase pairs of DNA, respectively. The chromosomal positions of the three types of deletions overlapped and together defined a continuous region of 27 kilobase pairs surrounding the prophage attachment site attPSPB. The 27-kilobase-pair segment contained no functions required for lytic growth of the phage, but DNA within this region was used as a template for RNA synthesis at several stages in the life cycle of SPB. In addition the transcription of DNA during lytic infection was found to be initiated over a large portion of one-half of the viral chromosome (the arbitrary left half). Subsequently, the synthesis of early RNA was terminated as late transcription continued on the opposite side of the chromosome. PMID- 2989559 TI - DNA-binding proteins present in varicella-zoster virus-infected cells. AB - DNA-binding proteins present in varicella-zoster virus-infected cells were identified by DNA-cellulose chromatography of radioactively labeled cell extracts. Seven virus-specific proteins, ranging in molecular weight from approximately 175,000 to 21,000, showed affinity for single- or double-stranded DNA or both. These proteins include the varicella-zoster virus major capsid protein, a phosphorylated tegument protein, and a 125,000-molecular-weight species which may be analogous to the major DNA-binding protein of herpes simplex virus. We also identified a number of DNA-binding phosphoproteins by these procedures. Finally, protein blot studies were carried out to determine whether these proteins bind preferentially to virus rather than to host cell DNA. PMID- 2989560 TI - Vesicular stomatitis virus NS proteins: structural similarity without extensive sequence homology. AB - The complete nucleotide sequence of the NS mRNA of vesicular stomatitis virus (New Jersey serotype) was established from two cDNA clones spanning the entire coding region of the mRNA. The gene is 856 nucleotides long and can code for a polypeptide of 274 amino acids. Comparison with the nucleotide sequence of the NS gene of the Indiana serotype revealed only 41% sequence homology. The deduced amino acid sequences of the NS proteins were only 32% homologous, with no identical stretches of more than five amino acids. However, at the C-terminal domain there was a conserved region of 21 amino acids with greater than 90% homology. Surprisingly, relative hydropathicity plots also demonstrated the presence of a large number of hydrophilic amino acids sequestered similarly over the N-terminal half of the protein. In addition, the total number of serine and threonine residues, presumptive phosphorylation sites, was similar and included seven serine and three threonine residues located at identical positions. It appears that during divergent evolution of these two vesicular stomatitis virus serotypes from a common ancestor, considerable mutation occurred in the main body of the gene but the overall structure of the protein was retained. The function of the NS protein in relation to the evolution of the two viruses is discussed. PMID- 2989562 TI - Truncated gag-related proteins are produced by large deletion mutants of Rous sarcoma virus and form virus particles. AB - Large deletion (LD) mutants of Prague strain Rous sarcoma virus subgroup B (PrB), derived by serial undiluted passage through chicken (C/E) cells, contain two deletions relative to wild-type virus. One of these joins gag sequences in the p12 coding region to env sequences in region encoding gp37; the other deletion spans the src region. Analysis of the viral proteins of QT6 cell clones containing only LD proviruses by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major truncated gag-related phosphoprotein of 60,000 to 66,000 daltons (P63LD). P63LD was stable, but could be cleaved in vitro to the predicted products by p15gag. A second gag-related LD protein of about 68,000 to 74,000 molecular weight (P70LD) was also found which often reacted with an anti gp37 serum. P70LD was unstable and may represent a short-lived gag-gp37 fusion protein. Finally, immunoprecipitation indicated that particles containing P63LD were shed from QT6-LD clones. Thin section preparations of these clones viewed in an electron microscope showed enveloped budding particles of "immature" morphology. Thus, the synthesis and release of particles from infected cells does not require cleavage of the gag precursor, nor does it require the presence of p15 or (most of) p12. PMID- 2989561 TI - Evolutionary variants of Rous sarcoma virus: large deletion mutants do not result from homologous recombination. AB - Large deletion (LD) mutants of Prague strain Rous sarcoma virus, subgroup B (PrB), derived by serial undiluted passage through chicken (C/E) cells, were isolated and characterized. Individual LD viruses were initially isolated by cloning in soft agar of infected, chemically transformed quail (QT6) cells. Two regions of the PrB genome were deleted in the formation of the LD virus. This resulted in the junction of gag sequences in p12 to env sequences in gp37, and in the loss of the src gene. DNA restriction analysis of biologically active lambda Charon 27-LD recombinant clones indicated that individual LD viruses contained similar but not identical deletion endpoints. Two LD isolates, LD25 and LD85, were further subcloned into pBR322, and the deletion junctions were examined by DNA sequencing. Although the gag-env deletion endpoints were identical in the two subclones, heterogeneity was observed across the src deletion in that both mutants analyzed had the same 5' endpoint but slightly different 3' endpoints. In all cases, only a single homologous base (always an A residue) was found at the deletion endpoint. S1 nuclease analysis of the RNA from a number of QT6-LD clones gave similar results, indicating that the LD population was composed of viruses with similar but not identical deletion endpoints. Such viruses may have been generated from errors during reverse transcription of the virion RNA with subsequent selection assuring their dominance in the population. PMID- 2989563 TI - Molecular cloning and physical mapping of the tupaia herpesvirus genome. AB - Purified virion DNA of about 200 kilobase pairs of tupaia herpesvirus strain 2 was cleaved with EcoRI or HindIII restriction endonuclease. Restriction fragments representing the complete viral genome including both termini were inserted into the EcoRI, HindIII, and EcoRI-HindIII sites of the bacterial plasmid pAT153. Restriction maps for the restriction endonucleases EcoRI and HindIII were constructed with data derived from Southern blot hybridizations of individual viral DNA fragments or cloned DNA fragments which were hybridized to either viral genome fragments or recombinant plasmids. The analysis revealed that the tupaia herpesvirus genome consists of a long unique sequence of 200 kilobase pairs and that inverted repeat DNA sequences of greater than 40 base pairs do not occur, in agreement with previous electron microscopic data. No DNA sequence homology was detectable between the tupaia herpesvirus DNA and the genome of murine cytomegalovirus, which was reported to have a similar structure. In addition, seven individual isolates of tupaia herpesvirus were characterized. The isolates can be grouped into five strains by their DNA cleavage patterns. PMID- 2989564 TI - Inguinal lymph node metastases from testicular tumor. AB - The Northern Israel Cancer Center serves 1 million inhabitants. Between 1968 and 1982, 33 patients with a diagnosis of nonseminomatous tumors of the testis were referred to this center. Of these patients inguinal lymph node metastasis developed in 4, each of whom had had risk factors for such metastasis. Two patients had undergone previous orchiopexy, and 2 had extension of the tumor to the epididymis and the tunica vaginalis testis. The inguinal region should be examined and watched carefully in patients with testicular tumor, especially those at high risk for inguinal metastases. Today, disease in patients with inguinal metastasis is curable by lymph node dissection and/or combination chemotherapy. PMID- 2989565 TI - Chlorpromazine: adjuvant therapy for the metabolic derangements created by urinary diversion through intestinal segments. AB - The hyperchloremic metabolic acidosis which can occur following urinary diversion through intestinal segments has been managed with bicarbonate or citrate salts. However, satisfactory management is not always possible with this form of treatment. The development of this acidosis has been attributed to intestinal reabsorption of urinary solutes or intestinal secretion of bicarbonate. Intestinal absorptive and secretory processes are modulated by an adenylate cyclase-cyclic AMP system. Chlorpromazine inhibits the effect of cyclic AMP on the intestinal mucosal cell. The use of chlorpromazine in the management of the hyperchloremic metabolic acidosis following urinary diversion was investigated. A canine model employing an ileal segment between ureter and bladder and a rat model in which urine is diverted through the entire colon have been developed. Chlorpromazine (5 mg./kg./day) was found to be efficacious in the management of the metabolic derangements that occur in both of these models. A case study is presented in which conventional management of this syndrome with bicarbonate salts was unsuccessful. The use of chlorpromazine as an adjuvant treatment allowed correction of the acidosis. PMID- 2989566 TI - Leads from the MMWR. Revision of case definition of acquired immunodeficiency syndrome for national reporting--United States. PMID- 2989567 TI - Leads from the MMWR. Results of human T-lymphotropic virus type III test kits reported from blood collection centers--United States, April 22 to May 19, 1985. PMID- 2989568 TI - Infection with HTLV-III/LAV and transfusion-associated acquired immunodeficiency syndrome. Serologic evidence of an association. AB - We studied patients with transfusion-associated acquired immunodeficiency syndrome (AIDS) and their blood donors for serologic evidence of infection with human T-cell lymphotropic virus type III/lymphadenopathy-associated virus with two enzyme-linked immunosorbent assays and a Western blot assay. All 19 patients with AIDS were seropositive by at least one test. In all 28 donor sets containing "high-risk" donors, at least one donor was seropositive by one or more tests. Of 255 donors not considered high risk, two (0.8%) were seropositive by all three tests. When 30 seropositive high-risk donors were evaluated a median of 29 months after donation, four (13%) had developed AIDS and eight (27%) had lymphadenopathy. Our findings support the hypothesis that human T-cell lymphotropic virus type III/lymphadenopathy-associated virus causes AIDS and indicate that seropositive high-risk donors may be at relatively high risk for developing AIDS or related conditions themselves. PMID- 2989569 TI - Revisiting the Holy Cross football team hepatitis outbreak (1969) by serological analysis. AB - Clinical and biochemical data collected during the Holy Cross College football team hepatitis A outbreak in 1969 suggested that 32 team members had icteric hepatitis, 58 had anicteric illness, and only seven were not infected. Using a currently available radioimmunoassay, we tested stored serum samples obtained during the outbreak for IgM antibody to hepatitis A virus (IgM anti-HAV). Only individuals with icteric hepatitis were found to have IgM anti-HAV in serum; those with presumed anicteric illness were shown not to be infected with hepatitis A virus. The attack rate was thus only 34%, not 93% as originally reported, and the incidence of icteric illness in those infected was 100%, not 33%. This serological analysis of a classic outbreak of hepatitis A illustrates the utility and importance of IgM anti-HAV testing in seroepidemiologic investigations of hepatitis outbreaks. PMID- 2989570 TI - Laboratory detection of marijuana use. AB - Frequent smoking of cannabis (marijuana) has been shown to be associated with a decline in social, mental, and perceptual skills and, during adolescence, with maladaptive emotional development. Urinalysis for the detection of such use can be a useful tool for the physician responsible for treatment and counseling of adolescents who develop habitual use of marijuana. Primary methods for urinalysis detection of cannabis use include the homogeneous enzyme immunoassay (EMIT) and the radioimmunoassay. These and other methods are discussed along with the issues of "false" results (both positive and negative) and the "limits of interpretation" that can be placed on a positive urine result. The pharmacokinetics and metabolism of the active constituents of cannabis are described as well as the interpretation of urinalysis results as they relate to use patterns. Guidelines are presented for the primary care physician for selecting candidates for such testing and for the use of such tests in the treatment or counseling of adolescents for whom marijuana abuse has become a psychological and social problem. PMID- 2989571 TI - Bowel function of healthy men consuming liquid diets with and without dietary fiber. AB - The importance of the level of dietary fiber intake on bowel function was measured in 16 healthy young males consuming self-selected and liquid enteral diets. Subjects consumed liquid Ensure as their sole nutrient source plus 0 (diet a), 30 (diet c), and 60 (diet d) g/day soy fiber, a fiber source high in hemicellulose that was added to the Ensure. Further, to examine whether heat processing affects the physiologic action of fiber, subjects consumed Enrich, an enteral formula similar to Ensure, which contains 30 g of the same soy fiber (diet b). The four diets were consumed in randomized order each for 10 days. Daily wet stool weight averaged 144.6 g on the self-selected diet and decreased significantly to 67.3 g when Ensure was consumed alone. Average stool weights on diets (b), (c), and (d) were 114.6, 100.2, and 150.3 g/day, respectively. Average fecal dry weights for the four diets were 19.1, 28.7, 25.0, and 30.3 g/day for (a d), respectively. Gastrointestinal transit time, as measured with radiopaque pellets, was longest on the Ensure diet, 72.4 hr, and approximately 2 days on the fiber-supplemented diets and the self-selected diet. Thus, soy fiber's physiologic effect on laxation was not changed by heat processing. Also, although stool weights were larger on higher fiber intakes, transit rates were similar on all the fiber-containing diets. PMID- 2989572 TI - [Passage into the human milk and clinical evaluation of sulbactam/cefoperazone]. AB - A combined agent of sulbactam, a beta-lactamase inhibitor, plus cefoperazone (SBT/CPZ: 1/1) was studied on the passage into the maternal milk as well on the clinical application and the results were followings: One or 2 grams of SBT/CPZ was given by one shot intravenous injection to puerperal mothers to determine the passage into the milk up to 6 hours after the administration. A trace of passage was detected for both SBT and CPZ as in the case of other antibiotics. Good clinical efficacy was observed in all 5 patients with female genital organ infections, to whom a daily dose of 2.0 g of SBT/CPZ was administered by drip infusion for 4 to 6 days. No side effects were noticed. PMID- 2989574 TI - Adult rat heart cell culture. The morphological study of tissue fragments on various environments in culture. AB - This paper reports a modification of Jacobson's method for culturing adult rat heart cells after mechanical and enzyme treatment. For heart cell dissociation, it is useful to wash tissue fragments 3 times with saline A solution after enzyme treatment, and it is important to change the medium immediately after culturing heart cells in order to prevent myocardial cells from degeneration caused by the enzyme solution and toxins in the cell debris. PMID- 2989573 TI - Effects of adenosine on transmembrane potential and sarcolemmal Na+-K+ ATPase activity of human atrial myocardium. AB - Adenosine effects on the transmembrane potential characteristics and the sarcolemmal Na+-K+ ATPase activity of human atrial myocardium were studied in tissue from 20 patients who were divided into 2 groups based on the maximum diastolic potentials (MDP) greater than or less than -60 mV. Group A consisted of 10 patients with MDP of 70.84 +/- 4.20 mV and Na+-K+ ATPase activity of 15.37 +/- 0.46 mumole Pi/mg/hr. Ten patients with MDP of 44.54 +/- 6.24 mV and Na+-K+ ATPase activity of 12.55 +/- 0.42 mumole Pi/mg/hr were included in group B. Adenosine had no effects on the electrophysiological properties and the sarcolemmal Na+-K+ ATPase activity of atrial myocardium at concentrations below 1 X 10(-5) M in either group. Adenosine resulted in mildly altered atrial transmembranes potentials without significant effect on Na+-K+ ATPase activity at concentrations between 1 X 10(-5) M and 5 X 10(-4) M. However, a significant reduction of transmembrane potentials and an apparent inhibition of Na+-K+ ATPase activity were observed only in tissue from group B. These results suggest that: 1) adenosine has no effect on the electrophysiological properties and the sarcolemmal Na+-K+ ATPase activity of human atrial myocardium at physiological concentrations; 2) adenosine induced inhibition of the sarcolemmal Na+-K+ ATPase activity in slow channel-dependent atrial tissues may be a mechanism responsible for the alterations of transmembrane potentials under unphysiological conditions; and 3) adenosine contributes to the genesis of cardiac arrhythmias during acute myocardial ischemia, which can reduce transmembrane potentials of the myocardial cells and may increase the myocardial adenosine level above its effective concentration. PMID- 2989575 TI - Calcium accumulating ability of mitochondria from bovine coronary artery. Comparison with aortic mitochondria. AB - The calcium accumulating ability of mitochondria isolated both from bovine coronary artery and aorta was investigated. Coronary artery and aorta were pretreated with 0.1% collagenase. Cytochrome c oxidase activities of mitochondria isolated from coronary artery and aorta showed 25-fold and 19-fold increases, respectively, as compared with those of each homogenate, whereas NADPH-cytochrome c reductase, potassium-phosphatase and Na+-K+ ATPase activities increased less than 2-fold. This suggests that the isolation procedure is capable of obtaining a subcellular fraction highly enriched with mitochondria. Mitochondrial calcium uptake activity of the coronary artery was approximately 250 nmoles Ca2+/mg protein/10 min, and was markedly depressed with metabolic inhibitors such as NaN3, ruthenium red and 2,4-dinitrophenol. Calcium uptake activity of bovine aortic mitochondria showed similar activity and a similar trend in sensitivity to metabolic inhibitors. By contrast, the onset of the calcium binding reaction of the aortic mitochondria was slower and the azide-sensitivity of the mitochondria to magnesium ATPase activity was lower than those for coronary artery mitochondria. The present study has provided a method for isolation of mitochondria with a high capacity of calcium uptake activity, which may prove meaningful for future physiological and pharmacological evaluation of mitochondrial calcium accumulation in vascular smooth muscle. PMID- 2989576 TI - Necropsy finding in a patient with apical hypertrophic cardiomyopathy. AB - Whether apical hypertrophic cardiomyopathy is a variant of classic hypertrophic cardiomyopathy or a separate entity is controversial. This is a case report of an apical hypertrophic cardiomyopathy. The patient was a 67-year-old man associated with giant negative T waves in electrocardiogram and asymmetric apical hypertrophy on echocardiogram. He died of liver cirrhosis and liver cell carcinoma. At necropsy the heart showed apical hypertrophy grossly and extensive disarray of myocardial fibers near the apex of the left ventricle histologically. The necropsy findings were indistinguishable from those of classic hypertrophic cardiomyopathy. This suggests that apical hypertrophic cardiomyopathy is a variant of hypertrophic cardiomyopathy. PMID- 2989577 TI - [Host factors in lung cancer patients--with reference to family and past histories]. AB - An analysis of cigarette smoking habits and family and past histories of 494 lung cancer patients was undertaken to determine whether host factors in such patients could be related to cancer. In men, 35.8% of the patients with squamous cell carcinoma had family histories of cancer, which is significantly higher than the ratio in control subjects (P less than 0.05). In women, 58.3% of the patients with bronchoalveolar cell carcinoma had family histories of cancer, and, in six out of eight of them, their parents had had the disease. As for past histories, 7.7% of the women with lung cancer had undergone surgery ovarian tumor. These results suggest that familial (hereditary) factors and predisposition play a role in the etiology of lung cancer. PMID- 2989578 TI - [Results of surgical treatment for large cell carcinoma of the lung]. AB - Twenty-six patients with histologically confirmed large cell carcinoma of the lung were evaluated. The overall five-year survival of these cases was 19.9%. Fifteen patients who had undergone surgical resection, excluding one operative death, had a five-year survival rate of 33.8%. None of the nonresected cases survived more than six months after the beginning of treatment. Among prognostic factors in surgical treatment of large cell carcinoma, survival correlated well with clinical stage, extent of lymph-node metastasis, curative resection and histological subtypes. As for histological subtypes, stratified subtype by division of nongiant cell carcinoma revealed poorer prognosis than that of nonstratified subtype and appeared to have some correlation with the frequency of vascular invasion by the tumor. From the clinical viewpoint, giant cell carcinoma should be regarded as a distinct clinical entity. Two cases, however, had benefit of long-term survival due to surgical modality. The clinicopathological analysis of those cases remains to be resolved in the future. PMID- 2989579 TI - [Retrospective study of short survival cases in hepatocellular carcinoma after transcatheter arterial embolization therapy]. AB - Twenty-two patients with hepatocellular carcinoma were treated with transcatheter arterial embolization therapy. In the six who died within about a month, serum albumin and Ch-E were lower, and total bilirubin and ICG R15 were higher than in the other cases. In four of them, more than 50% of the liver was occupied by tumor, and tumor thrombosis were found in the portal trunk or bilateral first portal branch. Five patients died of hepatic failure followed by upper gastrointestinal bleeding. One died of cachexia. The causes of short survival were 1) severe liver cirrhosis, 2) portal obstruction, 3) large tumor, 4) widespread TAE, 5) retrograde flow of gelfoam. PMID- 2989580 TI - [Nuclear magnetic resonance investigations of DMBA-induced rat breast cancers- the relationship between tumor doubling times and water proton spin-lattice relaxation times]. AB - Our studies of human neoplastic tissues had revealed that the T1 values of many malignant tissues had a tendency to be longer than those of normal ones. But the T1 values did not always reflect the grade of malignancy on account of the variety of their histologies. Therefore, by using DMBA-induced rat breast cancer, we investigated the relation between the degree of malignancy and the T1 values, and the following results were obtained. 1) The growth rate of the cancers had a correlation with their T1 values, and 2) the water content of the cancers correlated with their T1 values, but 3) there was no correlation between their growth rate and their water content. PMID- 2989581 TI - [Prostaglandins and related substances: biosynthesis]. PMID- 2989582 TI - [Physiological and pharmacological actions of prostaglandins and related substances on the respiratory system]. PMID- 2989583 TI - [Physiological and pharmacological actions of prostaglandins and related substances: the adipose tissue and prostaglandins]. PMID- 2989584 TI - [Physiological and pharmacological actions of prostaglandins and related substances: inflammation and allergy]. PMID- 2989585 TI - [Physiological and pharmacological actions of prostaglandins and related substances on mineral metabolism]. PMID- 2989586 TI - [Bronchial asthma and prostaglandin related substances]. PMID- 2989587 TI - [Prostaglandin-related substances and neoplasms]. PMID- 2989588 TI - [On the multivariate analysis of biochemical liver function tests in relation to histopathological diagnosis--with special reference to mass screening of hepatocellular carcinoma]. PMID- 2989589 TI - [One-step determination of guanase activity]. PMID- 2989590 TI - [Therapeutic effect of transarterial embolization on daughter nodules of hepatocellular carcinoma]. PMID- 2989591 TI - [Studies on platelet aggregation and cortinellus shiitake]. PMID- 2989592 TI - Establishment of Cockayne syndrome fibroblast cell line belonging to complementation group B by SV40 transformation. PMID- 2989593 TI - [Plasma angiotensin I converting enzyme and renin angiotensin aldosterone system in hypertensive subjects]. PMID- 2989594 TI - Peak systolic pressure-volume relationships in man: noninvasive determination by equilibrium gated radionuclide angiocardiography and cuff sphygmomanometry. PMID- 2989595 TI - [Designing and evaluation of a dynamic cardiac phantom for ECG gated blood-pool scintigraphy]. PMID- 2989596 TI - 2',3'-Cyclic nucleotide 3'-phosphohydrolase in acrylamide-induced optic neuropathy in rabbit. AB - The in vivo effects of acrylamide on the nerve tissues of the rabbit optic pathway were examined. The optic nerve of acrylamide-treated rabbits showed inflammatory changes and degeneration. The decrease of myelin was observed with myelin stain. The activity of 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC.3.1.4.37) which is firmly bound to myelin was markedly diminished by acrylamide intoxication. PMID- 2989597 TI - An analysis of precipitated withdrawal in rats acutely dependent on morphine. AB - Acute dependence on a single dose of morphine was assessed in Wistar rats by observing the frequencies of occurrence of several signs of withdrawal precipitated by naloxone, diprenorphine, Mr2097, Mr1452 and Mr2266. Naloxone significantly precipitated urination, paw shakes, head shakes and chewing. Diprenorphine significantly precipitated urination and chewing. Mr2097 precipitated urination, head shakes, teeth chattering and chewing. The selective kappa antagonists Mr1452 and Mr2266 significantly precipitated only urination and teeth chattering. Signs of the precipitated withdrawal by Mr2097 were mediated by stereoselective opioid receptors, as the other diastereoisomer, Mr2097, did not precipitate them. Stereospecific opioid receptors were also involved in the induction of acute dependence, as naloxone precipitated withdrawal only in I methadone-treated rats, but not in d-methadone treated rats. All the opioid antagonists produced at least some degree of "abstinoid" signs in morphine-free rats which might be caused by the blockade of endogenous opioids acting on mu and/or kappa receptors. The signs of withdrawal precipitated by naloxone and Mr2097 might be primarily mediated by mu receptors, those of diprenorphine by both mu and kappa receptors, and those by Mr1452 and Mr2266 were likely to be selectively mediated by kappa receptors. The latter aspect was further supported by experiments showing that the novel kappa agonist U-50488H did not precipitate withdrawal. A low degree of precipitation of withdrawal by Mr1452 and Mr2266 and the absence of precipitation of abstinence by U-50488H might be related to either a lack or an existence of a low proportion of kappa receptors in rat brain. Further experiments using selective agonists and antagonists are needed to evaluate these findings. PMID- 2989598 TI - Development of practical ELISA for detection of antibodies to bovine leukemia virus: a comparison of its sensitivity with that of virus neutralization and agar gel immunodiffusion tests. PMID- 2989599 TI - Reappearance of Getah virus infection among horses in Japan. PMID- 2989601 TI - Multiple cutaneous papillomas and carcinomas that develop spontaneously in a mouse mutant, the repeated epilation heterozygote Er/+. AB - After a chance observation that multiple cutaneous papillomas and squamous cell carcinomas occurred in 2 adult mice heterozygous for the repeated epilation gene Er, we surveyed a panel of 10 +/+ (wild type) and 30 Er/+ (heterozygous) mice from birth to over 2 years of age. Homozygous Er/Er mice could not be included since their defect is lethal at birth. Whereas no cutaneous tumors developed in the +/+ mice, 20 of the Er/+ mice, males and females, had developed 1-5 cutaneous papillomas and at least 1 cutaneous invasive squamous cell carcinoma by 2 years of age. No lesions were seen in mice younger than 6 months old. Although almost all Er/+ mice died with their tumor burden, no metastases have yet been proven histologically. The Er/+ mouse should serve as a useful model for the exploration of genetic factors in cutaneous squamous cell carcinomas in humans. PMID- 2989600 TI - Bovine papillomavirus type 1 monoclonal antibodies. AB - Bovine papillomavirus type 1 (BPV-1) and type 2 (BPV-2) are the etiologic agents of fibropapillomas in cattle. Polyclonal antisera produced against BPV-1 structural antigens are cross-reactive with BPV-2. In this study BPV-1 type specific monoclonal antibodies were produced that were not reactive with BPV-2. These monoclonal antibodies could be used for identification of BPV-1 structural antigens in acetone-fixed, frozen sections by immunofluorescence and Formalin fixed, paraffin-embedded sections by immunoperoxidase techniques. In addition, these antibodies could be used for identification and purification of BPV-1 virions by immune electron microscopy and immunoadsorption techniques, respectively. PMID- 2989602 TI - Establishment and characterization of the human cholangiocarcinoma cell line HChol-Y1 in a serum-free, chemically defined medium. AB - A human cholangiocarcinoma cell line, designated as HChol-Y1, was established in a protein-free, chemically defined medium after a very short period of primary culture in 0.1% fetal bovine serum (FBS)-containing medium. The cell line has been propagated in this medium for 2 years. The cells grew as a monolayer and the doubling time was about 52 hours. Addition of FBS did not stimulate cell growth (population-doubling time = 50 hr) or increase saturation density. The cells grown in a protein-free medium secreted small amounts of carcinoembryonic antigen (CEA) and large amounts of carbohydrate antigen (CA) 19/9 (CEA: 12.5 +/- 2.1 ng/10(6) cells/48 hr; CA 19/9: 760 +/- 52 IU/10(6) cells/48 hr); these tumor markers were immunohistochemically demonstrated in HChol-Y1 cells. Addition of FBS slightly stimulated the production of CEA and CA 19/9. The HChol-Y1 cell line was xenotransplantable in athymic nude mice and increased the serum CEA and CA 19/9 levels in the tumor-bearing nude mice. For determination as to whether a human carcinoma cell line can proliferate and secrete CEA and CA 19/9 in synthetic medium without any protein supplements, the cells were cultivated long term (2 yr) in a protein-free, chemically defined medium. When this method of cultivation is used, it is easy to purify these substances from spent medium, because contaminating antigens such as FBS or other substances usually added to cultures are absent. PMID- 2989603 TI - Chromosome 1 abnormalities: a common feature of pediatric solid tumors. AB - Abnormalities of chromosome 1 were found in 32 of 46 pediatric solid tumors including Ewing's sarcoma, Wilms' tumor, rhabdomyosarcoma, primitive neuroectodermal tumor, and hepatoblastoma. Trisomy of 1q was the most common abnormality, and breakpoints were most frequent in the region 1cen to 1p22. Abnormalities of chromosome 1 are not specific to any type of tumor. However, their frequent occurrence indicates that they may endow a clonal advantage in the development of cancer. PMID- 2989604 TI - Lymphokine-activated killer (LAK) cell phenomenon. IV. Lysis by LAK cell clones of fresh human tumor cells from autologous and multiple allogeneic tumors. AB - Activated killer cells are generated by the incubation of peripheral blood mononuclear leukocytes (PBL) in the lymphokine interleukin 2 (IL-2). Unseparated populations of these lymphokine-activated killer (LAK) cells lyse a variety of fresh noncultured human tumor targets, but they do not kill normal PBL. This study analyzed the generation and lytic specificity of LAK cell clones. Of 49 (84%) clones isolated by limiting-dilution techniques from a whole population of LAK cells, 41 manifested significant LAK cell activity. LAK cell clones had varied cell surface phenotypes. Clones with high and intermediate LAK cell activity were Leu 2+3-4+7-DR+Tac+ and Leu 2-3+4+7-DR+Tac+, respectively. Single LAK cell clones lysed multiple fresh human tumor targets including autologous sarcoma, 5 allogeneic sarcomas, and a colon cancer in addition to the cultured cell line K562. Autologous PBL were not lysed. Tumor targets were each lysed by multiple LAK cell clones. Sixteen subclones were derived from 5 of these LAK cell clones. These subclones had 99% or greater probability of being derived from a single cell. These subclones also exhibited lysis of multiple tumor targets. These findings suggest the existence of a shared determinant, expressed by multiple human tumors, which is recognized in common by multiple LAK cell clones. PMID- 2989605 TI - Cohort study of internal malignancy in genetic hemochromatosis and other chronic nonalcoholic liver diseases. AB - The risk of hepatocellular carcinoma (HCC) and other internal malignancies was examined in patients with genetic hemochromatosis (GH) by following 208 patients from the time of diagnosis to June 1983 and by comparing the numbers of cancers they developed with expected values constructed from cancer registry incidence data by means of actuarial methods. In addition, cancers occurring in a comparison group of 148 subjects with other chronic nonalcoholic liver diseases (CLD) were determined. Among the GH group, 16 new cases of HCC occurred subsequent to the diagnosis of GH, together with 8 other malignancies. The 16 cases of HCC reflect a 200-fold excess risk, which from all indications represents the first quantitation of the risk of this tumor in GH. There appears to be no increased risk of other malignancies in this disease. Among the CLD group only 1 HCC and 1 other malignancy occurred. PMID- 2989606 TI - Expression of collagenase IV (basement membrane collagenase) activity in murine tumor cell hybrids that differ in metastatic potential. AB - Expression of a basement membrane collagen-degrading metalloprotease activity (collagenase IV) was studied in a series of murine cell hybrids derived from fusions between highly metastatic cells (B16-F10RR) or moderately metastatic cells (UV-2237RR) and tumorigenic cells (K-1735 clone 16) or normal cells [peritoneal macrophages (PEC) or C3H mouse embryo fibroblasts (C3H-F)]. The collagenase IV activity of the parent cells and the hybrids was assayed in vitro and compared to the metastatic propensity of the same cells evaluated in both syngeneic (C57BL/6 X C3H/HeN)F1 mice and BALB/c nude mice. The level of collagenase IV activity secreted by the parent lines correlated with their metastatic capacity. The highly metastatic B16-F10RR line secreted the highest enzyme activity, whereas the tumorigenic but nonmetastatic K-1735 clone 16 and the normal parents PEC and C3H-F secreted the lowest enzyme activity. The enzyme activity was completely inhibited with EDTA. The hybrid derived from fusion of cells from two metastatic cell lines as well as hybrids derived from a metastatic and a nonmetastatic tumor cell line expressed higher levels of collagenase IV activity than either parent, and this expression was associated with a high ability to produce metastases in both nude and syngeneic mice. Fusion of metastatic cells with normal cells produced hybrid cells that exhibited suppression of both collagenase IV activity and metastatic capacity. Collagenase IV activity and metastatic propensity can, therefore, be altered by somatic cell hybridization; in the series of hybrids examined in these experiments the expression of type IV collagen-degrading metalloprotease activity and the metastatic ability were closely correlated, which suggests that collagenase IV activity and other properties required for metastasis are genetically linked. PMID- 2989607 TI - Atrial natriuretic factor: structure and functional properties. PMID- 2989609 TI - [Multiple primary malignant tumors of the respiratory organs (review of the literature)]. PMID- 2989608 TI - Primary acquired hypoaldosteronism. PMID- 2989610 TI - [Level of plasma ACTH and aldosterone in endotoxic shock in rats]. PMID- 2989611 TI - [Effect of lithium on the proliferation of fibroblasts and tumor cell lines in vitro]. AB - The specificity of a proliferation-inducing effect of lithium was investigated. Cell lines of embryonal and adult solid tumors as well as fibroblasts were cultured in lithium-concentrations ranging from 0,5 to 5,0 mmol/l. Neuroblastoma cell-lines SK-N-SH, SK-N-LO, IMR 32, osteosarcoma-cell-line SAOS 2, melanoma-line IgR 3 and a fibroblast-line were used in this study. Cell proliferation was measured with a 3H-TdR-incorporation-assay and a tumor-stem cell-assay, the fibroblast-proliferation was measured following growth as monolayer respectively. No stimulation of proliferation was observed. The data of this in vitro study are basic for the clinical evaluation of the benefit of lithium in attenuation of chemotherapy induced leukopenia in patients with solid tumors. PMID- 2989613 TI - Inconsistent stimulation of plasma ACTH through corticotropin-releasing factor in a patient with central Cushing's disease due to pituitary adenoma. AB - Three uncommon findings were observed in a case of Cushing's disease due to macroadenoma: no suppression of plasma ACTH during an 8-mg dexamethasone test, a negative corticotropin-releasing factor test, and a normal X-ray of the sella turcica. Despite these findings, the diagnosis of pituitary was confirmed Cushing's syndrome by computerized tomography and a transphenoidal operation. PMID- 2989612 TI - Rare association of herpes simplex virus IgM-specific antibodies and Guillain Barre syndrome successfully treated with plasma exchange and immunosuppression. AB - Herpes simplex virus (HSV) has been associated with various neurological disorders. In contrast, HSV infection is very rarely found in acute polyneuroradiculitis. In this report, a patient is described with a severe course of Guillain-Barr? syndrome (GBS). HSV IgM-specific antibodies and a rise of complement-fixation antibodies were detected. During the acute phase of neurologic syndrome, a nerve biopsy showed myelin damage and IgM deposits on the inner layer of the perineurium. Plasma exchange, in combination with immunosuppression, was successfully applied as a treatment in the relapsing course of GBS. Finally, after recovery, HSV-specific IgM antibodies disappeared. PMID- 2989614 TI - Comparison of serum angiotensin-converting enzyme in Graves' disease, toxic nodular goiter, and other thyroid conditions. AB - Angiotensin-converting enzyme (ACE) was measured in plasma of patients with Graves' disease and with toxic nodular goiter, as well as in their treated counterparts and in normal controls. A significant elevation of ACE levels and a positive correlation between ACE, thyroxine, and triiodothyronine levels was found in both groups of thyrotoxicosis. Parallel fluctuations of triiodothyronine and ACE levels, albeit with a certain lag of the latter, was observed in patients in whom multiple measurements were taken. ACE levels, which were also determined in hypothyroid patients and in patients with euthyroid goiter on suppression therapy, were in the same range as normal controls and as in treated thyrotoxic patients. We conclude that the pathogenesis of thyrotoxicosis does not play a role in ACE elevation but that increased thyroid hormone seems to induce elevation of ACE. ACE elevation in thyrotoxicosis may constitute an integral part of the renin-angiotensin axis. PMID- 2989615 TI - Cyanamide, ethanol, and liver cell inclusion bodies. PMID- 2989616 TI - Quantification of expired metabolites following potassium cyanide administration: a new method. AB - A sensitive and specific radiolabel method for measuring expired hydrogen cyanide (HCN) or carbon dioxide (CO2) derived from cyanide was developed. An ethanol collecting solution containing a 10(-2) M of cobalt chloride trapped 88% of the H14CN passed through the solution following acid volatilization of a known amount of K14CN. The range of linearity, r = 0.998, exceeded the 0.01 to 0.1 mumoles tested for measuring the pulmonary metabolites. The cobalt chloride collecting solution trapped less than 0.02% of the 10 to 100 mumoles of 14CO2 generated by acid hydrolysis of 14C-sodium bicarbonate (NaH14CO3). Introduction of a second collecting solution specific for CO2, composed of ethanol: ethanolamine (2:1, v/v), was used to collect the CO2 derived from cyanide. Following the subcutaneous (sc) administration of 4.6 mg/kg of potassium cyanide (KCN) and 4.5 microcuries (microCi) K14CN, 1 to 2% and 2 to 3% of the dose was expired as H14CN and 14CO2, respectively. PMID- 2989617 TI - Physical separation of aortic corticoid receptors with type I and type II specificities. AB - Previous gel filtration binding assay studies indicated that rat vascular smooth muscle cells contained corticoid receptor I and corticoid receptor II sites which could be distinguished on the basis of their relative affinities for aldosterone and dexamethasone. Ion-exchange chromatography experiments were designed to separate the two sites for further studies on their physical characteristics and role in vascular smooth muscle cell physiology. Cultured aortic cells were incubated with 5-10 nM 3H steroid alone or in the presence of 10-fold non radioactive steroid competitor for 30 min at 37 degrees C. Following cell lysis, total cellular protein-bound steroid was isolated using Sephadex G-25 and applied to a DEAE-cellulose ion-exchange column. Three peaks of radioactivity were eluted using a 1-200 mM sodium phosphate gradient: peak I (30-38 mM), peak II (52-64 mM), and peak III (92-102 mM). Peaks I and II contained 60% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor II sites (dexamethasone greater than aldosterone). Peak III contained 40% of the eluted radioactivity and exhibited the same steroid specificity as corticoid receptor I sites (aldosterone greater than dexamethasone). These studies support the binding assay data on steroid specificity and relative proportion of type I and II sites. They also document the existence of type I and II corticoid receptors with different physicochemical characteristics in rat aortic smooth muscle cells. PMID- 2989618 TI - 17 alpha-oxidoreductase activity in kidneys of male and female mice of various ages. AB - Male and female mice at 0-120 days of age were used. Homogenates of kidneys were incubated with [14C]4-androstene-3,17-dione, and 17 alpha-oxidoreductase activity per g tissue was examined. The activities of 17 alpha-oxidoreductase in the kidneys of both sexes increased markedly with age during sexual development by up to 150-fold and reached the maximum values (2700 and 1500 nmol/g/h in male and female kidneys, respectively) at 60 days of age. In the adult male mouse kidney, the activity in isolated cortex fractions was 14 times as high as the activity in isolated medulla fractions; in the medulla fractions renal tubules from the cortex accounted for 3-15% of the total tissue. Furthermore, histochemical examination showed the activity present only in the cortex, at which much higher levels in the tubules than in the glomerulus. Activity at 35-120 days of age was significantly higher in male kidneys than in female kidneys. The difference appears to be induced by testicular androgens during sexual development, since neonatal castration in males resulted in decreases of activity to levels similar to those in female kidneys. However, castration at 60 days of age showed no significant effect on the activity. The present results show that the activity per g tissue of 17 alpha-oxidoreductase in the mouse kidney increases markedly with age, and that the activity is largely confined to the renal tubules of the cortex. PMID- 2989619 TI - Bronchial carcinoid arising in intralobar bronchopulmonary sequestration with vascular supply from the left gastric artery. Case report. AB - We report the unique case of a large, nonmetastasizing bronchial carcinoid tumor that arose within an intralobar bronchopulmonary sequestration in a 45-year-old man. The vascular supply to the sequestrated area within the left lower lobe as well as to the carcinoid tumor originated from atypical branches of the left gastric artery and the thoracic aorta. A left lower lobe lobectomy was performed. Histologically, a typical carcinoid tumor without lymph node metastases was found (T2 N0 M0). Seven years postoperatively, the patient is without signs of recurrence. PMID- 2989620 TI - Prevention of free radical-induced myocardial reperfusion injury with allopurinol. AB - Growing evidence supports the concept that oxygen free radicals are an important cause of myocardial ischemic and reperfusion injury. This study was designed to determine if toxic oxygen metabolites may exacerbate ischemic injury upon reoxygenation. Left ventricular function was studied in a group of seven dogs receiving intermittent, 4 degrees C, hyperosmolar, hyperkalemic (KCI 25 mEq/L) saline cardioplegic solution. This group was compared to a group (n = 7) receiving a hyperkalemic (KCI 25 mEq/L) cardioplegic solution designed to scavenge superoxide anion and hydroxyl radical: superoxide dismutase (3,000 U/ml) and mannitol (325 mOsm/L). A third group of five animals received allopurinol pretreatment (50 mg/kg/day) for 72 hours and hyperkalemic saline cardioplegic solution. After 60 minutes of ischemia (10 degrees to 15 degrees C) and 45 minutes of reperfusion, left ventricular mechanical function was better in the groups receiving free radical scavengers and allopurinol pretreatment than in the group receiving only hyperkalemic saline cardioplegic solution. Free radical scavengers preserved myocardial function in this model of hypothermic global ischemia and reperfusion. Our data support the concept that injury occurs primarily during reperfusion with the generation of oxygen free radicals via the hypoxanthine-xanthine oxidase reaction. Allopurinol has potential clinical application in the prevention of reperfusion injury. PMID- 2989621 TI - Viruses as risk factors or causes of human leukaemias and lymphomas? PMID- 2989623 TI - Animal models: lessons from feline and bovine leukaemia virus infections. PMID- 2989622 TI - Epidemiology of human retroviruses. PMID- 2989624 TI - The epidemiology of lymphomas and leukaemias in Africa--an overview. PMID- 2989626 TI - Microcomputer-assisted densitometer for quantitative receptor autoradiography. AB - We describe here a simple, inexpensive microcomputer-assisted densitometer for use with quantitative receptor autoradiography. The resolution of this system is approximately 100 micron. With this system, and an accompanying program DENSIT, it is relatively easy to convert density values of autoradiograms into molar quantities of bound ligand. The general design of the system and the logic of the DENSIT program are applicable to a variety of hardware systems. PMID- 2989625 TI - Influence of life-style on the pattern of leukaemia and lymphoma subtypes among Nigerians. PMID- 2989627 TI - [Nursing of a dying patient. A thought on nursing after interactions with a patient in the terminal stage of liver cancer]. PMID- 2989628 TI - [Vitamin D. II. Clinico-therapeutic implications]. PMID- 2989629 TI - [Liver transplantations at the Huddinge Hospital--experience, indications, needs]. PMID- 2989630 TI - [Ectopic internal carotid artery in the tympanic cavity. Intraoperative complications and their treatment. A case report]. AB - Ectopy of the internal carotid artery involving the middle ear occurs only rarely. In this case, middle ear surgery will almost always result in severe haemorrhage. After package of the external auditory canal and the epipharynx, immediate closure or ligation of the internal carotid artery is mandatory. The authors describe the interventional neuroradiological treatment as an alternative way to the known neurosurgical methods like cervical and intracranial ligation of the artery. Intravasal blocking of the leakage was done by using a detachable balloon filled with contrast medium. During this procedure cortical evoked potentials were measured showing no decrease of the homolateral haemispheric blood flow. Although this procedure cannot maintain the continuity of the internal carotid artery, it preserves a maximum of collateral vessels. The cortical evoked potentials simultaneously show the brain function during blockage of the artery. PMID- 2989631 TI - Fluoxetine, a selective inhibitor of serotonin uptake, potentiates morphine analgesia without altering its discriminative stimulus properties or affinity for opioid receptors. AB - The analgesic effect of morphine in the rat tail jerk assay was enhanced by the serotonin uptake inhibitor, fluoxetine. Tail jerk latency was not affected by fluoxetine alone. Morphine's affinity for opioid receptors labeled in vitro with 3H-naloxone or 3H-D-Ala2-D-Leu5-enkephalin was not altered by fluoxetine, which has no affinity for these sites at concentrations as high as 1000 nM. In rats trained to discriminate morphine from saline, fluoxetine at doses of 5 or 10 mg/kg were recognized as saline. Increasing the fluoxetine dose to 20 mg/kg did not result in generalization to either saline or morphine. The dose response curve for morphine generalization was not significantly altered by fluoxetine doses of 5 or 10 mg/kg. Those rats treated with the combination of morphine and 20 mg/kg of fluoxetine did not exhibit saline or morphine appropriate responding. Fluoxetine potentiates the analgesic properties of morphine without enhancing its affinity for opioid receptors or its discriminative stimulus properties. PMID- 2989633 TI - Daily rhythms of benzodiazepine receptor numbers in frontal lobe and cerebellum of the rat. AB - Behavioural, biochemical and neurophysiological evidence suggests that gamma aminobutyric acid (GABA) may play an important role in the neural control of circadian rhythms. Central receptors for benzodiazepines are functionally coupled to GABA receptors and appear to mediate behavioural effects of exogenous benzodiazepines. The binding of 3H-flunitrazepam to synaptic plasma membranes prepared from various regions of rat brain was examined at 6-hour intervals over a 36-hour period. Prominent daily rhythms in receptor number (Bmax) were observed in the frontal lobe and the cerebellum but not in the temporoparietal regions, hypothalamus or medulla/pons. Binding was highest during periods of sleep/low activity with a significant decrease occurring just prior to waking. These results suggest that daily fluctuations in benzodiazepine receptor numbers may be related to the temporal control of sleep/wake and muscle activity cycles. PMID- 2989632 TI - Increased analgesic potency of morphine and increased brain opioid binding sites in the rat following chronic naltrexone treatment. AB - Implantation of rats with prolonged-release naltrexone pellets increased both morphine's analgesic potency in the tailflick assay and radiolabeled opioid binding in the brain. The increases in both radiolabeled opioid binding and morphine potency were time-dependent. Implantation for 24 hours did not increase binding, whereas increases of approximately 45% were seen following 8 days of implantation. Similarly, morphine's analgesic potency, measured as ED50 values, was increased by 50% following 8 days of exposure to naltrexone while a 24 hour exposure had no significant effect. PMID- 2989634 TI - Caffeine tolerance: behavioral, electrophysiological and neurochemical evidence. AB - The development of tolerance to the stimulatory action of caffeine upon mesencephalic reticular neurons and upon spontaneous locomotor activity was evaluated in rats after two weeks of chronic exposure to low doses of caffeine (5 10 mg/kg/day via their drinking water). These doses are achievable through dietary intake of caffeine-containing beverages in man. Concomitant measurement of [3H]-CHA binding in the mesencephalic reticular formation was also carried out in order to explore the neurochemical basis of the development of tolerance. Caffeine, 2.5 mg/kg i.v., markedly increased the firing rate of reticular neurons in caffeine naive rats but failed to modify the neuronal activity in a group exposed chronically to low doses of caffeine. In addition, in spontaneous locomotor activity studies, our data show a distinct shift to the right of the caffeine dose-response curve in caffeine pretreated rats. These results clearly indicate that tolerance develops to the stimulatory action of caffeine upon the reticular formation at the single neuronal activity level as well as upon spontaneous locomotor activity. Furthermore, in chronically caffeine exposed rats, an increase in the number of binding sites for [3H]-CHA was observed in reticular formation membranes without any change in receptor affinity. We propose, therefore, that up-regulation of adenosine receptors may underlie the development of tolerance to the CNS effects of caffeine. PMID- 2989635 TI - Glucocorticoid receptor regulation. AB - Glucocorticoids, like other classes of steroid hormones, must bind to cellular receptors in order to exert their effects. Because of this central role in mediating hormone action, it is important to elucidate those factors that control receptor content. The purpose of this minireview is to summarize the recent work that explores the mechanisms through which cells modulate their glucocorticoid receptor binding capacity. PMID- 2989636 TI - Differential mediation of the anticonvulsant effects of carbamazepine and diazepam. AB - Possible mechanisms of action of carbamazepine and diazepam on amygdala-kindled seizures were studied using compounds acting at the central and "peripheral-type" benzodiazepine binding sites. Ro-15-1788, a selective antagonist at the central benzodiazepine site, blocked the anticonvulsant effect of diazepam, but not of carbamazepine. In contrast, Ro5-4864, which acts at the "peripheral-type" benzodiazepine site, blocked the anticonvulsant effect of carbamazepine, but not of diazepam. The effect of Ro5-4864 was itself reversed by PK-11195, a compound that displaces Ro5-4864 binding in vitro and in vivo. These data indicate that the anticonvulsant effects of carbamazepine and diazepam on amygdala-kindled seizures are differentially mediated and suggest that the "peripheral-type" benzodiazepine binding site is functionally involved in the anticonvulsant effect of carbamazepine. PMID- 2989637 TI - Absent aldosterone response to metoclopramide in patients with high spinal cord transection: evidence that metoclopramide stimulates aldosterone secretion through central pathways. AB - This study evaluates dopaminergic regulation of aldosterone secretion in 6 patients with high spinal cord transections. Administration of the dopamine antagonist metoclopramide resulted in a marked rise in plasma aldosterone and 18 hydroxycorticosterone levels in 12 normal individuals, but no change in plasma levels of these zona glomerulosa corticosteroid products in spinal cord patients. Spinal cord transected patients also did not have the rise in plasma renin activity that was observed in normals following metoclopramide administration. Basal levels of aldosterone, 18 hydroxycorticosterone, corticosterone and renin activity as well as the aldosterone responses to graded dose infusion of adrenocorticotropin were similar in the spinal cord patients and the normals. These data suggest that dopaminergic regulation of adrenal zona glomerulosa corticosteroid and renal renin secretion is absent in patients with high spinal cord transections, suggesting that intact neural pathways from the central nervous system are necessary for metoclopramide stimulation of aldosterone and renin secretion in men. Since basal plasma aldosterone levels were normal in spinal cord transected patients, it appears that the absence of dopaminergic control does not result in elevated secretion. PMID- 2989638 TI - Rabbit anti-idiotypic antibodies raised against monoclonal anti-morphine IgG block mu & delta opiate receptor sites. AB - Rabbit antibodies have been raised against murine monoclonal anti-morphine Fab fragments. Following affinity purification, these antibodies competitively inhibit morphine binding to anti-morphine monoclonal antibodies, yet do not recognize normal mouse IgGs, suggesting that the antibodies produced are anti idiotypic with respect to the anti-morphine IgG. More importantly, the purified antibodies competitively inhibit binding of morphine, naloxone, and D-ala-2-D-leu 5-enkephalin to rat brain opiate receptors. PMID- 2989639 TI - [3H] diazepam binding to human granulocytes. AB - [3H]-diazepam binds to sites on human granulocyte membranes, with little or no binding to platelets or lymphocytes. These [3H]-diazepam binding sites are of the peripheral type, being strongly inhibited by R05-4864 (Ki = 6.23nM) but only weakly by clonazepam (Ki = 14 microM). Binding of [3H] diazepam at 0 degree is saturable, specific and stereoselective. Scatchard analysis indicates a single class of sites with Bmax of 109 +/- 17f moles per mg of protein and KD of 3.07 +/ 0.53nM. Hill plots of saturation experiments gave straight lines with a mean Hill coefficient of 1.03 +/- 0.014. Binding is time dependent and reversible and it varies linearly with granulocyte protein concentration over the range 0.025 0.300 mg of protein. PMID- 2989641 TI - 1,25-Dihydroxyvitamin D3 regulates proliferation of activated T-lymphocyte subsets. AB - The biologically active vitamin D metabolite, 1,25-(OH) 2D3 suppressed phytohaemagglutinin (PHA)-induced lymphocyte proliferation dose-dependently (0.1 nM-100 nM), and decreased the OKT4+/OKT8+ ratio and transferrin-receptor-positive (OKT9+) cells. A possible parallelism between expression of 1,25-(OH) 2D3 receptors and interleukin 2 (IL2)-receptors recognized by anti-Tac antibody was not confirmed in this study. However, the addition of exogenous IL2 abolished the inhibitory effects of 1,25-(OH) 2D3 on PHA-stimulated T-cell proliferation, and the decrease of OKT4+ and OKT9+ T-cell in this population. Among various vitamin D3 analogues examined, 1,25-(OH) 2D3 was the most potent anti-proliferative effect, followed in order by 1,24S-(OH) 2D3, 1 alpha OH D3, 25 OH D3 and 24,25 (OH) 2D3. PMID- 2989640 TI - Regulation of adenosine 3':5'-monophosphate-dependent protein kinase in cerebral cortex. AB - Alterations in the cyclic AMP-dependent protein kinase activity ratio in response to putative neurotransmitters and other cyclic AMP-elevating agents in intact cerebral cortical slices and Krebs-Ringer particulate preparations from cerebral cortex were examined. Both norepinephrine (30 microM) and forskolin (20 microM) produced a time-dependent increase in intracellular levels of cyclic AMP in cerebral cortical slices which was paralleled by an increase in both cyclic AMP and the protein kinase activity ratio. The increases were maximal at 5 min. and remained elevated for at least 15 min. Forskolin, norepinephrine, adenosine and isoproterenol produced a concentration-dependent increase in both cyclic AMP and the protein kinase activity ratio, however, the degree of increase observed was dissimilar. Thus, a 5-fold change in intracellular cyclic AMP resulted in only a 2-fold increase in the activity ratio. Of the agents examined, forskolin produced the most marked change in the activity ratio (from 0.23 to 0.78 at 100 microM) while isoproterenol at 100 microM produced only a 50% increase in the activity ratio. The half-time for the decline in forskolin elicited elevations of either the activity ratio or cyclic AMP was about 4-6 min. In the presence of the phosphodiesterase inhibitor, Ro 20-1724, both were significantly prolonged being 60-70% of the maximum observed immediately after forskolin stimulation, at 15 min. Potentiation of forskolin elicited increases in the activity ratio by Ro 20 1724 were also observed but the increase in the activity ratio was maximal at 7.5 min. while cyclic AMP accumulations continued to rise during the entire 15 min. incubation. Particulate preparations from cerebral cortex were found to contain a cyclic AMP-dependent protein kinase which could be activated 2 to 3-fold with either forskolin, norepinephrine, or adenosine. Unlike the intact brain slice the changes in protein kinase activity ratio and intracellular levels of cyclic AMP in cell-free particulate preparations were similar in both time and degree. PMID- 2989642 TI - Long-term changes in self-stimulation threshold by repeated morphine and naloxone treatment. AB - To analyse the interaction between endogenous opioid systems and brain reward, the influence of repeated treatment for 3 weeks with morphine and the opioid antagonist naloxone was investigated in rats with self-stimulation electrodes in the ventral tegmental area. Changes in threshold of self-stimulation determined by a response rate insensitive two lever method were considered as changes in reward. Morphine induced a temporary decrease of the response rate which lasted 3 days, and decreased the threshold for self-stimulation. The effect on threshold remained present till morphine treatment was discontinued, indicating that tolerance does not develop to this effect of morphine. Repeated naloxone treatment gradually increased the threshold for self-stimulation. This effect persisted after discontinuation of naloxone treatment. It is concluded that blockade of opioid receptors induces long term changes in the setpoint of self stimulation reward. PMID- 2989643 TI - GABA in morphine analgesia and tolerance. AB - Drugs affecting various steps of GABA transmission exhibit analgesia in a variety of experimental models in animals; this analgesic response generally requires high doses of the drugs and does not appear to be opiate-like since the GABAergic analgesia is naloxone-insensitive and lacks dependence liability. The outcome of the analgesia response is variable when opiate and GABAergic drugs are administered together; however, directly acting GABA receptor stimulants and GABA transaminase inhibitors generally enhance the analgesic effect of opiates. The development of newer GABAergic drugs with greater potency and specificity may offer an alternative to opiate analgesics. The results obtained over the years, on the possible involvement of the GABA system in morphine tolerance and dependence are equivocal. Studies on region-specific changes in opiate-GABA interaction as well as opiate-GABA-benzodiazepine interaction are needed to further elucidate the role of GABA on opiate system. PMID- 2989644 TI - Modulation of erythrocyte Na transport pathway(s) by excess Na intake. AB - Different Na transport pathways were studied in the erythrocytes of 10 normotensive subjects who received 240 meq/day of Na in excess of their usual diet. In most of these subjects the maximal rate (Vmax) of the Na,K pump and the Na,K-cotransport system was markedly decreased on the first day of the diet. In some of these subjects, excess Na intake induced an increase in the apparent affinity for internal Na for the Na,K pump and the Na,K-cotransport system. The decrease in the Na,K pump fluxes was not concomitant to that of the co-transport system and not accompanied with an increase in blood pressure or cation concentration in the plasma. Interestingly, the apparent affinity for internal Li of the Li-Na exchange was markedly increased without alteration of the Vmax. The passive permeability for Na and the cellular Na content were not altered by excess Na intake. Ouabain and bumetanide at low concentrations respectively induced an increase in the apparent affinity for internal Na of the Na,K pump and the Na,K- cotransport system. These results are similar to those observed after excess Na intake. These later agree with the hypothesis that Na homeostasis regulates some endogenous factors with ouabain-like and furosemide-like properties that might contribute to the regulation of cellular Na handling. PMID- 2989645 TI - Reduction of GABAB receptor binding induced by climbing fiber degeneration in the rat cerebellum. AB - When the rat cerebellar climbing fibers degenerated, as induced by lesioning the inferior olive with 3-acetylpyridine (3-AP), GABAB receptor binding determined with 3H-(+/-)baclofen was reduced in the cerebellum but not in the cerebral cortex of rats. Computer analysis of saturation data revealed two components of the binding sites, and indicated that decrease of the binding in the cerebellum was due to reduction in receptor density, mainly of the high-affinity sites, the Bmax of which was reduced to one-third that in the control animals. In vitro treatment with 3-AP, of the membranes prepared from either the cerebellum or the cerebral cortex, induced no alteration in the binding sites, thereby indicating that the alteration of GABAB sites induced by in vivo treatment with 3-AP is not due to a direct action of 3-AP on the receptor. GABAA and benzodiazepine receptor binding labelled with 3H-muscimol and 3H-diazepam, respectively, in both of brain regions was not affected by destruction of the inferior olive. These results provide evidence that some of the GABAB sites but neither GABAA nor benzodiazepine receptors in the cerebellum are located at the climbing fiber terminals. PMID- 2989646 TI - Inhibition of Purkinje cell firing by systemic administration of phenylisopropyl adenosine: effect of central noradrenaline depletion by DSP4. AB - The effect of the metabolically stable adenosine analog (-)-N6(R-phenyl isopropyl)-adenosine (PIA) on the rate of spontaneous Purkinje cell firing was studied in anesthetized rats. In control animals, systemically administered PIA elicited only small and inconsistent changes in firing rate. However, in animals previously treated with DSP4 (50 mg/kg i.p.), which selectively lesions central noradrenergic afferents, or with the adrenergic antagonist sotalol (15 mg/kg), PIA elicited consistent decreases in firing rate. These effects were antagonized by the systemic administration of the adenosine receptor antagonist aminophylline (50-150 mumol/kg). Local administration of adenosine by pressure ejection caused a dose-dependent depression of Purkinje cell firing that was likewise inhibited by the methylxanthine. In DSP4 treated rats the depression of synaptic transmission by adenosine in rat hippocampus in vitro was unaltered, and theophylline did not cause any marked rise in Purkinje cell firing, suggesting that DSP4 does not sensitize neurons to the depressant effects of adenosine derivatives. PIA also caused a dose-dependent decrease in arterial blood pressure and a decrease in heart rate that was of equal magnitude in control and DSP4 treated rats. The results show that the central effects of systemically administered adenosine analogs are altered by procedures that disrupt the normal depressant effect of tonic noradrenergic input. PMID- 2989647 TI - Absorption, metabolism, and excretion studies of carbon 14- and tritium-labeled derivatives of a ketomethylene containing tripeptide. AB - Tritium and Carbon 14 analogs of the angiotensin converting enzyme inhibitor ketoACE were synthesized and their oral absorption, metabolism and excretion in rats were investigated. KetoACE, a ketomethylene analog of the tripeptide Bz-Phe Gly-Pro, was slowly absorbed at a 35% level upon oral administration. It is rapidly eliminated from the blood with a half-life of about 10 minutes. Its excretion is primarily via the bile duct and it is excreted as 80% unchanged drug. The only identified metabolite consisting of 5-10% of the excreted radioactivity was determined to be the reduced ketoACE in which the ketone group was reduced to a hydroxyl. PMID- 2989649 TI - Modulation of the cardiac membrane-bound cyclic nucleotide phosphodiesterase: inhibition due to a contamination of TLC purified S-adenosyl-L-[methyl-3H] methionine by Zn++ ions. AB - Cardiac membranes pretreated with S-Adenosyl-L-[methyl-3H] methionine([3H] SAM) purified on TLC silica gel 60 F254 plates exhibited a marked decrease in cyclic AMP and cyclic GMP phosphodiesterase activity. However, this inhibition did not appear when membranes were incubated with either [14C] SAM or unlabelled SAM. We showed that, during the TLC purification of [3H] SAM, which involved an acidic elution step, minute amounts of the fluorescent indicator F254 (Zn sulfur) were eluted. The contaminating Zn++ ions strongly inhibited cyclic nucleotide phosphodiesterase activity and phospholipid methylation with I50 values in the micromolar range. PMID- 2989648 TI - Intact brain cells: a novel model system for studying opioid receptor binding. AB - This paper presents the use of a novel tissue preparation to study opioid receptor binding in viable intact cells derived from whole brains of adult rats. Mechanically dissociated and sieved cells, which were not homogenized at any stage of the experimental protocol, and iso-osmotic physiological buffer were used in these experiments. This system was adapted in order to avoid mechanical and chemical disruption of cell membranes, cytoskeletal ultrastructure or receptor topography by homogenization or by the use of non-physiological buffers, and to mimic in vivo binding conditions as much as possible. Using [3H]naloxone as the radioligand, our studies showed saturable and stereospecific high-affinity binding of this opioid antagonist in intact cells, which in turn showed consistently high viability. [3H]Naloxone binding was also linear over a wide range of tissue concentrations. This technique provides a very promising model for future studies of the binding of opioids and of many other classes of drugs to brain tissue receptors in a more physiologically relevant system than those commonly used to date. PMID- 2989650 TI - Benzodiazepine receptor binding in vivo with [3H]-Ro 15-1788. AB - In vivo benzodiazepine receptor binding has generally been studied by "ex vivo" techniques. In this investigation, we identify the conditions where [3H]-Ro 15 1788 labels benzodiazepine receptors by true "in vivo" binding, i.e. where workable specific to nonspecific ratios are obtained in intact tissues without homogenization or washing. [3H]-Flunitrazepam and [3H]-clonazepam did not exhibit useful in vivo receptor binding. PMID- 2989651 TI - Temperature-induced phase change in a fat. A study by electron spin resonance. AB - Electron Spin Resonance, Differential Scanning Calorimetry and rheological techniques have been used to study the physical changes induced by temperature in lard and in the solid and liquid fractions obtained by fractionation of lard at 15 C. The mobilization process of a C18 fatty acid nitroxide derivative dispersed in the molten fat has been observed in the temperature range -50 to +70C. The mobilization of the probe seemed to be concomitant with the melting of the low melting point glycerides. Above this temperature, all the probes were in the liquid phase and their mobility reflected the viscosity of their liquid environment, or the viscosity of the bulk fat when crystal was no longer present. Probe mobility was temperature dependent, and it was identical for the three fats at the same temperature, despite their different triglyceride compositions. PMID- 2989652 TI - Stimulation by lipoxygenase products of superoxide anion production in FMLP treated neutrophils. AB - Our recent observation that leukotriene B4 (10(-9)M) is a potent enhancer of FMLP initiated neutrophil superoxide anion formation prompted an evaluation of the ability of other lipoxygenase products and related compounds to modulate this response. The results indicate that FMLP-evoked O-2 may be enhanced by 10(-8)-10( 7)M levels of a number of lipids, in addition to LTB4, including 5-HPETE, 5-HETE, 5,15-DiHPETE and by higher levels of other 15-series lipoxygenase products and arachidonic acid. It is of interest that the relative potency of these agents in potentiating the superoxide response to FMLP approximately parallels their reported ability to induce chemotactic activity in leukocytes. PMID- 2989653 TI - Discordance of plasma DHEA-S, DHEA, and cortisol responses with various ACTH regimens. AB - The origin and time course of ACTH-stimulated adrenal androgens are controversial. Since previous protocols have used differing ACTH preparations and routes of administration, we sought to compare plasma DHEA and DHEA-S responses to four ACTH regimens. Fourteen young men underwent the following five three-day study periods: (1) placebo, (2) alpha 1-24 ACTH, 400 micrograms intravenously (IV); (3) alpha 1-39 ACTH, 400 micrograms intramuscularly (IM); (4) alpha 1-18 (D Ser1, Lys17, Lys18) ACTH, 400 micrograms IM; and (5) alpha 1-18 ACTH, 400 micrograms IV. alpha 1-18 ACTH IV had the most potent and prolonged corticotropic effect, listing more than 24 hours, as measured by plasma cortisol, 17-OHCS, and cortisol secretion rates. alpha 1-39 ACTH and alpha 1-18 ACTH IM were corticotropic up to 12 hours, and alpha 1-24 ACTH IV was corticotropic only up to one hour. Plasma DHEA rose acutely at one hour with all of the ACTH regimens (P less than 0.05). This response was correlated with cortisol (r = 0.62, P less than 0.01) and reflected the relative corticotropic potency of each of the ACTH regimens. Plasma DHEA-S, on the other hand, did not rise acutely at one hour with any of the regimens. DHEA-S did rise at 12 hours with three of the ACTH regimens (alpha-1-24, alpha 1-39, alpha 1-18 IM), but this response was not synchronous with cortisol (r = 0.14, P = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989654 TI - Altered erythrocyte Na-K pump in anorectic patients. AB - The status of the erythrocyte sodium pump was evaluated in a group of patients suffering from anorexia nervosa and a group of healthy female control subjects. Anorectic patients showed significantly higher mean values of digoxin-binding sites/cell (ie, the number of Na-K-ATPase units) with respect to control subjects while no differences were found in the specific 86Rb uptake (which reflects the Na-K-ATPase activity) between the two groups. A significant correlation was found between relative weight and the number of Na-K-ATPase pump units (r = -0.66; P less than 0.0001). Anorectic patients showed lower serum T3 concentrations (71.3 +/- 53 ng/dL) with respect to control subjects (100.8 +/- 4.7 ng/dL; P less than 0.0005) and a significant negative correlation between T3 levels and the number of pump units (r = -0.52; P less than 0.003) was found. Our study therefore shows that the erythrocyte Na-K pump may be altered in several anorectic patients. We suggest that this feature could be interrelated with the degree of underweight and/or malnutrition. PMID- 2989655 TI - Transposon mutagenesis and genetic mapping of the rglA and rglB loci of Escherichia coli. AB - The rglA and rglB genes code for two different proteins which cleave the hydroxymethylated cytosine residues of T-even phages. We isolated Tn10 and Tn5 insertion mutants of the above genes and of the genes in and around the rglA and rglB loci. These insertions were used to construct a detailed genetic map. Our results show that the rglA gene maps at 25.24 min and the rglB gene at 98.39 min on the standard Escherichia coli K12 genetic map. PMID- 2989656 TI - Developmental control of transduced dopa decarboxylase genes in D. melanogaster. AB - Seventeen new euchromatic integration sites of the dopa-decarboxylase gene (Ddc) have been generated using p-mediated transduction. The developmental expression of the integrated genes was examined by monitoring the embryonic induction of dopa decarboxylase enzyme activity (DDC) and by monitoring the developmental pattern of DDC activity from late third instar to eclosion. The majority of inserts are regulated correctly within about 30% of controls. Several cases of multiple insertion events were recovered and these show correspondingly elevated levels of activity and are regulated normally. The pattern of expression of one insert (15C) falls outside the normal range. Multiple copies of transduced Ddc genes are used to test for effects of elevated gene dose on levels of expression. One insert on the X chromosome shows little or no dosage compensation. Possible reasons for the differences between the regulation of transduced genes in Drosophila and the regulation of transformed genes in mammalian systems are discussed. PMID- 2989657 TI - Genome organization in Halobacterium halobium: a 70 kb island of more (AT) rich DNA in the chromosome. AB - The more A + T rich fractionated component (FII DNA) of the Halobacterium halobium genome constitutes one third of the total DNA and upon isolation consists of covalently closed circular DNA (pHH1 and minor cccDNA) and nonsupercoiled sequences. We have investigated the physical organization of the non cccDNA in FII by a chromosome walk using one copy of the halobacterial insertion element ISH1 as a start point. This chromosome walk led to the isolation of 160 kb of chromosomal DNA containing 70 kb of FII DNA covalently linked to more G + C rich sequences (FI DNA). Copies of three previously characterized insertion elements (ISH1, ISH2, and ISH26) as well as at least 10 other repeated sequences are clustered within this chromosomal FII DNA "island". Unique sequences are found in the FI DNA flanking the FII DNA island as well as in 40 kb of FI DNA surrounding the bacterio-opsin gene. The presence of pHH1 in H. halobium and closely related species correlates with the occurrence of the characterized chromosomal FII DNA island. Halophilic purple membrane producing isolates YC81819-9, GN101, SB3 and GRA lack pHH1 and the 70 kb FII DNA, but contain all of the FI DNA sequences tested. We propose that pHH1 and this chromosomal FII DNA are characteristic genomic components of H. halobium and closely related species, and, that the 70 kb FII DNA might represent a large insertion in the chromosome of H. halobium and closely related species. The conservation of both FI and FII DNA sequences can be used for strain classification and determination of evolutionary relationships among halo bacteria. PMID- 2989658 TI - The novel gene(s) ARD of plasmid pKM101: alleviation of EcoK restriction. AB - The host-controlled K restriction of unmodified phage lambda was 10-100-fold alleviated in the wild-type strain E. coli K12, carrying plasmid pKM101 of incompability group N. pKM101-mediated release of K restriction was also observed in lexA-, recA-, and recB- strains of E. coli K12. By restriction mapping Tn5 insertions in pKM101, which reduced pKM101-mediated alleviation of restriction, were shown to be located within the BglIIB fragment approximately 11 kb anticlockwise from the RI site of pKM101. We have termed the gene(s) promoting the alleviation of K restriction of phage lambda ard (alleviation of restriction of DNA). It was shown (1) that ard function affected only the EcoK restriction system and not the EcoB, EcoRI, EcoRIII, or EcoPI systems, (2) ard gene(s) did not mediate EcoK type modification of lambda DNA and did not increase the modification activity of the EcoK system in a way similar to that observed with gene ral of bacteriophage lambda. PMID- 2989659 TI - Passive immunization of mice with monoclonal antibodies to glycoprotein gB of herpes simplex virus. AB - To investigate the protective ability of monoclonal antibodies (MCAs) to viral glycoprotein in herpes simplex virus (HSV) infection, athymic nude mice were inoculated intracutaneously with HSV type-1 (HSV-1) in the midflank. Three hours after inoculation, one group of mice was passively immunized with one of a series of MCAs to glycoprotein gB of HSV-1, and a control group of mice was given phosphate buffered saline alone. The control mice died within 16 days after infection, whereas the mice passively immunized with any of the MCA showed suppressed development of skin lesions. Three of six mice given MCA failed to develop any visible lesions and no HSV could be isolated from the lumbar dorsal root ganglia of these mice 60 days after the challenge. BALB/c mice were also protected from infection with HSV type 2 by passive immunization with MCA to HSV 1 gB. PMID- 2989660 TI - Conversion of Staphylococcus aureus cells to stable L-forms in a liquid growth medium. PMID- 2989661 TI - Conjugation of Sendai virus with pullulan and immunopotency of the conjugated virus. PMID- 2989662 TI - Activation of human monocytes in leprosy. AB - In leprosy, the common etiologic agent is the same Mycobacterium leprae, but the clinical manifestations are various, including the tuberculoid and lepromatous types. In tuberculoid type leprosy, macrophages in the granuloma differentiate into epithelioid cells; in the lepromatous type, in contrast, they differentiate into lepra cells containing multiple M. leprae. Thus host factors, which regulate macrophage activities, determine the type of leprosy. To understand such regulation of macrophage activities, we assayed superoxide production, hydrogen peroxide production and glucose consumption in monocytes in vitro. Glucose consumption spontaneously increased, with lymphokine enhancing the consumption rate. Superoxide production increased spontaneously and decreased from the 4th day; lymphokine added on the 4th day supressed the decrease of superoxide production. Hydrogen peroxide production increased until the 3rd day of culture. Twenty-four hour incubation with lymphokine, from day 0 to the 1st day, had no effect on hydrogen peroxide production, while from the 2nd to 3rd day it enhanced such production. Supernatants of lymphocytes incubated with M. leprae were prepared from tuberculoid and lepromatous patients. Tuberculoid supernatant enhanced reactive oxygen production and glucose consumption, while that from lepromatous patients had no remarkable effect on glucose consumption or reactive oxygen production. The range of spontaneous increase and decrease of reactive oxygen production was greater than the regulatory effect of lymphokine on these activities. These data show that rapid provision of new monocytes to the granuloma is one of the important factors in the defense mechanism, that lymphocytes separated from lepromatous patients are not activated in response to M. leprae antigen, and that they do not secrete corresponding lymphokines. PMID- 2989663 TI - [Effect of aspartate amino acids on aspartokinase activity of oligotrophic bacteria]. AB - The object of this work was to study the effect of aspartate amino acids taken separately or in combinations on the aspartokinase activity of Hyphomicrobium and Methylobacterium methylotrophous strains. Aspartokinase was shown to be a polyvalent enzyme regulated by the coordinated action of two amino acid pairs: lysine+threonine and threonine+methionine. PMID- 2989664 TI - Analysis of world-wide age-specific data for the association of cervical cancer with infections by herpes simplex virus type 2. AB - World-wide incidence data obtained from the scientific literature are used to expand to age-specific incidence rates the observation that the incidence of invasive cervical cancer in many human populations is related to the incidence of infections with herpes simplex virus type 2 (HSV-2) in the same community. The data for the incidence of the tumor were obtained from international registries. Data for prevalence of HSV-2 antibodies were used to evaluate the incidence of past primary infections with the virus and a model was used to generate the required age-specific data. Given the nature of the data used and the assumptions implied by the analysis, the conclusions remain speculative. They nevertheless constitute the best available assessment of the age distributions of cancer cases related and unrelated to the virus, and of the time elapsing from the initial virus infection to the development of an invasive tumor. PMID- 2989665 TI - Sensory symptoms in Parkinson's disease and the opioid system. AB - Primary sensory symptoms not related to somatic disease, are often experienced by patients with Parkinson's disease. These subjective sensations may either accompany or precede the motor disorder. This paper provides evidence supporting a role for the endogenous opioid system in the pathogenesis of sensory symptoms in Parkinson's disease. PMID- 2989666 TI - Use of heat-treated clotting-factor concentrates in patients with haemophilia and a high exposure to HTLV-III. AB - In a group of 126 Australian patients with haemophilia, who were receiving lyophilized clotting-factor concentrates prepared from locally collected plasma, a high prevalence of antibody to human T-cell lymphotropic virus III (HTLV-III) was demonstrated in those with severe disease. Patients with moderate or mild disease had a much lower prevalence of HTLV-III antibody. After heat treatment of lyophilized factor VIII and factor IX concentrates (60 degrees C for 72 hours) to inactivate the virus, the losses of activity of an intermediate-purity and of a fibrinogen-poor factor VIII concentrate, and of the coagulant activity of a factor IX concentrate, were within acceptable limits. The solubility of the intermediate-purity factor VIII concentrate was markedly decreased; the fibrinogen-poor factor VIII concentrate and the factor IX concentrate were readily soluble. In-vivo recovery and survival of heated concentrates were equivalent to those of the unheated products, and they were effective in the treatment of spontaneous and traumatic haemorrhages. PMID- 2989667 TI - First International Conference on the Acquired Immunodeficiency Syndrome. PMID- 2989668 TI - [Computerized tomography in the study of pleural diseases]. PMID- 2989669 TI - Effective cisplatin (DDP) based chemotherapy in the treatment of hepatoblastoma. AB - Nine of 11 patients with hepatoblastoma treated with cisplatin (DDP) based chemotherapy had a complete (CR) or partial (PR) remission. Five of these patients had measurable pulmonary disease and four achieved a CR of pulmonary lesions. The average interval of disease control following DDP was three times that of Adriamycin (ADR). DDP is an effective agent in the treatment of hepatoblastoma. PMID- 2989670 TI - Dental enamel as an in vivo radiation dosimeter. AB - The determination of the radiation exposure history of the population has become increasingly important in the study of the effects of low-level radiation. The present work was started to try to obtain an in vivo dosimeter that could give an indication of radiation exposure. Dental enamel is the only living tissue which retains indefinitely its radiation history, and electron spin resonance measurements have shown that the radiation signal can be resolved down to about 10 cGy. Measurements on samples from the general population give radiation exposure estimates that are reasonable, and one measurement on a patient who had radiotherapy to the mouth area showed a good correlation with tumor dose. We believe that this is an important new indicator of radiation dose and taken together with exposure histories should provide important data for epidemiological studies as well as accidental exposures. PMID- 2989671 TI - Multiregulatory element of filamentous bacteriophages. PMID- 2989672 TI - Coagulase-negative staphylococci and the epidemiological typing of Staphylococcus epidermidis. PMID- 2989675 TI - Influence of digoxin and diuretic therapy on sweat fluid composition. AB - The effect of digitalis and diuretic therapy on sweat fluid composition was investigated. Patients treated for congestive heart failure with a combination of digoxin and diuretics demonstrated a higher concentration of sodium and chloride in their sweat fluid when compared to age-matched controls. The administration of diuretics alone did not affect sweat fluid composition. The digoxin-induced increase in sodium concentration was significantly higher than that observed for chloride. These data suggest that digoxin markedly inhibits sodium reabsorption along the sweat gland tubule, whereas chloride transport is affected to a lesser degree. A significant correlation between sweat fluid sodium and serum digoxin concentrations was observed. We conclude that the increased digoxin-induced sweat sodium and chloride losses may affect sodium homeostasis in patients with congestive heart failure. PMID- 2989676 TI - An evaluation of apatite-based pulp capping materials. PMID- 2989673 TI - In bacteria which grow on simple reductants, generation of a proton gradient involves extracytoplasmic oxidation of substrate. PMID- 2989677 TI - Revision of the case definition of acquired immunodeficiency syndrome for national reporting--United States. PMID- 2989678 TI - Results of human T-lymphotropic virus type III test kits reported from blood collection centers--United States, April 22,-May 19, 1985. PMID- 2989679 TI - Cytomegalovirus infection and hemolytic anemia due to glucose-6-phosphate dehydrogenase deficiency. PMID- 2989680 TI - Participation of active oxygen species in the induction of chromosomal aberrations by cadmium chloride in cultured Chinese hamster cells. AB - The effect of various scavengers of active oxygen species on the induction of chromosomal aberrations by cadmium chloride (CdCl2) was investigated in cultured Chinese hamster V79 cells. Incidences of chromosomal aberrations by CdCl2 were partially or fully reduced by the presence of catalase, mannitol (a scavenger of hydroxyl radicals) and butylated hydroxytoluene (BHT, an antioxidant). These findings may indicate participation of the active oxygen species such as hydrogen peroxide (H2O2) or hydroxyl radicals in the clastogenicity of cadmium. In contrast, superoxide dismutase (SOD) and dimethylfuran (a scavenger of singlet oxygen) did not influence incidences of chromosomal aberrations by CdCl2. These results suggest that superoxide anion and singlet oxygen are not directly involved in the clastogenicity of the metal. The presence of aminotriazole (an inhibitor of catalase) increased incidences of chromosomal aberrations by CdCl2. This emphasizes participation of H2O2 in the clastogenicity of cadmium. PMID- 2989681 TI - Study of the mutagenic activity of 6 hepatotoxic pharmaceutical drugs in the Salmonella typhimurium microsome test, and the HGPRT and Na+/K+ ATPase system in cultured mammalian cells. AB - Several pharmaceutical drugs show strong hepatotoxicity during therapeutic use. We have studied 6 of them: aminophenazone, clofibrate, nifuroxazide, oxamniquine, perhexiline maleate, tienilic acid. Their mutagenicity was assessed in the Ames test on 6 strains of Salmonella typhimurium, and in V79 Chinese hamster lung cells using a rat-hepatocyte-mediated metabolic activation system and the HGPRT and Na+/K+ ATPase assay. Nifuroxazide was positive in the Ames test in two Salmonella strains (TA100, and TA100 Fr1). In the hepatocyte-mediated mammalian V79 cell system, nifuroxazide, clofibrate and aminophenazone were negative; oxamniquine and tienilic acid were positive with and without metabolic activation in tests looking for ouabain and 6-thioguanine resistance. Perhexiline maleate was negative for the direct induction of 6-thioguanine resistance without metabolic activation, and positive after metabolisation mediated by primary rat's hepatocytes. These results suggest the need for some caution in the use of some pharmaceutical drugs because of hepatotoxicity and because 3 out of 6 drugs were shown to be slightly mutagenic in mammalian cells. PMID- 2989682 TI - [Experimental renal infection with Aspergillus fumigatus in mice: a chemotherapeutic model of an organic mycosis]. PMID- 2989683 TI - Kinetoplast DNA from Trypanosoma vivax and T. congolense. AB - We have analysed kinetoplast DNA (kDNA) of the African trypanosomes Trypanosoma vivax and T. congolense. The maxi-circles from these organisms resemble those of T. brucei in size, but only to a limited extent in sequence as judged from restriction enzyme digests and DNA X DNA hybridization. The kDNA networks of T. vivax have three distinguishing features: they contain the highest maxi-circle concentration of any kDNA (at least twice that of T. brucei); they contain the smallest mini-circles (465 bp) yet found thus far and the width of the kDNA nucleoid in thin sections is correspondingly small (55 nm against 91 nm for T. brucei); they contain a substantial fraction of mini-circle dimers. PMID- 2989684 TI - Identification of mitochondrial genes in Trypanosoma brucei and homology to cytochrome c oxidase II in two different reading frames. AB - We have determined the nucleotide sequence of a 3.3 kilobase segment of the kDNA maxicircle of Trypanosoma brucei brucei 164. The nucleotide sequence and its predicted translated sequence have homology to cytochrome c oxidase subunits I and II (CO I and II) and mammalian unidentified reading frame 1 (URF 1). Amino acid homology to CO II extends for 170 residues from the amino terminus in one reading frame and then continues in another reading frame for 39 residues to the carboxyl terminus. Similar results have been obtained for Leishmania tarentolae [de la Cruz, V.F., Neckelmann, N. and Simpson, L. (1984) J. Biol. Chem., in press] and T. brucei 427 [Hensgens, L.A.M., Brackenhoff, J., De Vries, B.F., Sloof, P., Tromp, M.C., Van Boom, J.H. and Benne, R. (1984) Nucleic Acids Res. 12, 7327-7344]. This may indicate that novel events are required for expression of this gene. Amino acid homology to URF 1 exists predominantly at the amino terminal end although no corresponding AUG codon occurs in this area. An alternative initiation codon may therefore be utilized by trypanosome mitochondria. Two other open reading frames (ORFs) were detected and these are discussed with reference to transcripts from this region. ORFs corresponding to transcripts are organized compactly and are distributed more equally on both strands compared to ORFs of other mitochondrial systems. PMID- 2989674 TI - Revised genetic linkage map of Bacillus subtilis. PMID- 2989685 TI - Lymphadenopathy-associated virus isolated from bronchoalveolar lavage fluid in AIDS-related complex with lymphoid interstitial pneumonitis. PMID- 2989686 TI - Late-onset adrenal hyperplasia in hirsutism. AB - We studied the incidence of late-onset adrenal hyperplasia as a cause of hirsutism, its association with the major histocompatibility complex, and its clinical expression. Twenty-four of 400 women seen because of hirsutism were found to have late-onset adrenal hyperplasia, diagnosed on the basis of a high plasma level of 17-hydroxyprogesterone, and its marked increase after ACTH stimulation. The degree of hirsutism varied widely. Plasma antigen levels were high, especially the level of androstenedione, whereas 5 alpha-reductase activity, considered to be a good index of peripheral androgen utilization, showed frequent normal or low values. The 24 patients were genotyped, along with 84 family members, and plasma hormones were measured in the family members. We found a high correlation between late-onset adrenal hyperplasia and HLA antigens B14 and Aw33. Similar biologic profiles were observed in the patients and those of their siblings who were HLA identical (n = 9), confirming that late-onset adrenal hyperplasia is linked to the histocompatibility complex. These nine siblings had no hirsutism. We therefore conclude that the role of skin sensitivity to androgens is important in determining the clinical expression of this disorder. PMID- 2989687 TI - Familial pyrimidinemia and pyrimidinuria associated with severe fluorouracil toxicity. PMID- 2989688 TI - Isolation of LAV/HTLV-III from a patient with amyotrophic lateral sclerosis. PMID- 2989689 TI - Effect of naloxone on food consumption in obesity. PMID- 2989690 TI - Orchestration at the synapse. PMID- 2989691 TI - Oncogenes homologous to steroid receptors? PMID- 2989692 TI - Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. AB - Human chronic myelogenous leukaemia is characterized by a reciprocal translocation between chromosomes 9 and 22 resulting in an abbreviated form of chromosome 22 and the transfer of the abl cellular oncogene from chromosome 9 into the bcr gene of chromosome 22. Characterization of an 8-kilobase RNA specific to chronic myelogenous leukaemia shows it to be a fused transcript of the two genes. The fused protein that would be produced is probably involved in the malignant process. PMID- 2989693 TI - Increased MHC H-2K gene transcription in cultured mouse embryo cells after adenovirus infection. AB - The class I major histocompatibility complex (MHC) antigens are highly polymorphic cell-surface proteins whose expression is essential for the cellular immune response against virus-infected, abnormal and foreign cells. Transformation of primary rat cell cultures by the oncogenic adenovirus 12 (Ad12) results in suppression of the transplantation antigens, thus enabling the transformed cells to escape the immune response and efficiently form tumours in vivo. In contrast, transformation of the same cells with the non-oncogenic adenovirus 5 (Ad5) does not suppress the transplantation antigens and, consequently, they elicit an effective (MHC-restricted) immune response. Here, however, we show that infection of mouse embryo cells with both viruses initially increases the level of transcripts from the H-2Kb transplantation antigen gene. Both the adenovirus E1a (12S RNA) and E1b genes are required for activation of the H-2K gene and measurement of the relative rate of transcription indicates that the increase in the level of H-2K messenger RNA following infection is at least in part due to a gene-specific transcriptional activation. The newly transcribed H-2Kb mRNA is then properly transported to the cytoplasm. PMID- 2989694 TI - Production of human alpha-interferon in silkworm using a baculovirus vector. AB - Microorganisms are generally used for mass production of foreign gene products, but multicellular organisms such as plants have been proposed as an economical alternative. The silkworm may be useful in this context as it can be cultured easily and at low cost. We have therefore developed a virus vector to introduce foreign genes, for example, the gene for human alpha-interferon (IFN-alpha), into silkworms. We used the baculovirus Bombyx mori nuclear polyhedrosis virus (BmNPV) which has a large (greater than 100 kilobases, kb) double-stranded circular DNA genome within its rod-shaped capsid. Baculoviruses have been used previously as vectors for expression of beta-interferon and beta-galactosidase in established cell lines. Although BmNPV has not been used previously as an expression vector, it has an advantage over the baculovirus Autographa californica NPV in that it has a narrower host range and will not grow in wild insect pests in the field. In the present study, the polyhedrin gene encoding the major inclusion body protein of BmNPV was identified by hybridization with complementary DNA and cloned in a plasmid. For insertion of foreign genes, we constructed a recombinant plasmid carrying a polylinker linked to the promoter of the polyhedrin gene, and inserted the IFN-alpha gene into this plasmid. The resulting plasmid and the BmNPV genomic DNA were co-transfected into BM-N cells, and stable recombinant viruses isolated by plaque assay on BM-N cells. The recombinant virus replicated in silkworm larvae, which synthesized as much as 5 X 10(7) units (approximately 50 micrograms) of interferon in their haemolymph. PMID- 2989695 TI - Retrovirus-induced de novo methylation of flanking host sequences correlates with gene inactivity. AB - The pattern of DNA methylation changes during development of eukaryotes, and hypomethylation frequently correlates with gene expression (for reviews see refs 1-4). A causal relationship between hypermethylation and gene inactivity has been established for retroviral genomes which are methylated de novo when inserted into the germ line of mice (ref. 5; for review, see ref. 6). The mutual interaction of the provirus with the host genome can influence virus expression and can result in inactivation of the host gene by insertional mutagenesis. We report here that the insertion of a provirus can change the methylation pattern of the host DNA. Sequences flanking the provirus become methylated de novo within 1 kilobase (kb) of the integration site. In Mov-13 mice, which carry a lethal mutation of the alpha 1(I) collagen gene, de novo methylation of host DNA is associated with a change in chromatin conformation. This suggests that virus induced DNA methylation can alter DNA-protein interactions and thereby interfere with correct gene activation during embryonic development. PMID- 2989696 TI - A viral enhancer element specifically active in human haematopoietic cells. AB - One particular class of DNA regulatory elements, the enhancers or activators, can, relatively independently of distance and orientation, dramatically increase the transcriptional activity of homologous and heterologous promoters located in cis (see refs 1-3 for reviews, also refs 4-6). Sequence differences between various heterologous enhancers may explain their apparent host- and/or tissue specific action. Furthermore, differences in the transcriptional control elements may contribute to viral tropism. At least for murine leukaemia virus isolates, thymotropism and leukaemogenicity have been attributed to alterations within the viral long terminal repeat, which harbours their enhancers and other transcriptional control elements. We report here the identification of a viral enhancer element possessing a very restricted tissue range. The enhancer is active in all human cells of the haematopoetic system tested, but not in cells of fibroblast or epithelial origin. PMID- 2989697 TI - Safety cabinets and AIDS. PMID- 2989698 TI - Cloning, sequence and expression of two distinct human interleukin-1 complementary DNAs. AB - Two distinct but distantly related complementary DNAs encoding proteins sharing human interleukin-1 (IL-1) activity (termed IL-1 alpha and IL-1 beta), were isolated from a macrophage cDNA library. The primary translation products of the genes are 271 and 269 amino acids long, although expression in Escherichia coli of the carboxy-terminal 159 and 153 amino acids produces IL-1 biological activity. PMID- 2989699 TI - Human T-cell lymphotropic virus type-I antibodies in Falashas and other ethnic groups in Israel. AB - Epidemiological studies of the human T-cell leukaemia/lymphoma virus type I (HTLV I), a type-C retrovirus of the human T-lymphotropic virus family, have used serological surveys to identify population subgroups possessing a high prevalence of naturally occurring HTLV-I-specific antibodies. Studies carried out to delineate the global distribution of the virus have demonstrated natural antibodies to HTLV-I in the serum of healthy donors from specific geographical areas, and have defined viral endemic areas in Japan, the Caribbean basin, Africa and the southeastern United States. Such studies have suggested that the prevalence of HTLV-I antibodies is directly correlated with age, is associated with the clinical syndrome of adult T-cell lymphoma, and is associated with transmission from mother to child. A separate subtype of the human retrovirus, HTLV-II (refs 21, 22), has also been identified. The population of Israel in part comprises groups of immigrants of various ethnic and geographical origins. Because of this, and the fact that Israel has a highly developed public health system, we surmised that the ethnic groups in Israel could be used in a seroepidemiological survey of HTLV infection. The serological survey reported here demonstrates a high prevalence of HTLV-I antibodies in new immigrants from Ethiopia. This previously ethnically and geographically isolated group, the 'Black Jews' or 'Falashas', from the Gondar region in the northern rural highlands of Ethiopia, has the highest endemic rate of HTLV-I yet reported outside Japan. PMID- 2989700 TI - Expression of active human clotting factor IX from recombinant DNA clones in mammalian cells. AB - Haemophilia B, or Christmas disease, is an inherited X-chromosome-linked bleeding disorder caused by a defect in clotting factor IX and occurs in about 1 in 30,000 males in the United Kingdom. Injection of factor IX concentrate obtained from blood donors allows most patients to be successfully managed. However, because of impurities in the factor IX concentrate presently in use, this treatment involves some risk of infection by blood-borne viruses such as non-A, non-B hepatitis and the virus causing acquired immune deficiency syndrome (AIDS). Because of the recent concern about the increasing incidence of AIDS amongst haemophiliacs, a factor IX preparation derived from a source other than blood is desirable. Here, we report that after introduction of human factor IX DNA clones into a rat hepatoma cell line using recombinant DNA methods, we were able to isolate small amounts of biologically active human factor IX. PMID- 2989701 TI - Lattice mobility and anomalous temperature factor behaviour in cytochrome c'. AB - Atomic temperature factors (B-values) obtained from X-ray refinement experiments provide empirical estimates of protein mobility that have been correlated with both theoretical simulations of protein dynamics and experimental studies of antibody reactivity. The comparison of B-values with protein solution properties requires adjustment of the apparent atomic mobilities to compensate for the effects of the crystal environment. Here we compare crystallographically independent subunits of the dimeric cytochrome c' from the bacterium Rhodospirillum molischianum to examine how lattice effects influence refined B values. In addition to local effects on protein mobility at crystal contacts, we show that B-value differences up to 12 A between subunits result from lattice disordering effects that approximate to concerted rotations of the molecules about a crystal symmetry axis. PMID- 2989702 TI - Amino-acid substitutions at codon 13 of the N-ras oncogene in human acute myeloid leukaemia. AB - DNAs from four out of five patients with acute myeloid leukaemia (AML) tested by an in vivo selection assay in nude mice using transfected mouse NIH 3T3 cells were found to contain an activated N-ras oncogene. Using a set of synthetic oligonucleotide probes, we have detected a mutation at codon 13 in all four genes. The same codon is mutated in an additional AML DNA that is positive in the focus-formation assay on 3T3 cells. DNA from the peripheral blood of one patient in remission does not contain a codon 13 mutation. PMID- 2989703 TI - Structural organization of the bcr gene and its role in the Ph' translocation. AB - The Philadelphia (Ph') chromosome, an abnormal chromosome 22 (ref. 1), is one of the best-known examples of a specific human chromosomal abnormality strongly associated with one form of human leukaemia, chronic myelocytic leukaemia (CML). The finding that a small region of chromosome 9 which includes the c-abl oncogene is translocated to chromosome 22 prompted studies to elucidate the molecular mechanisms involved in this disease. We have demonstrated previously that the chromosome 9 of one patient with CML contains a breakpoint 14 kilobases (kb) 5' of the most 5' v-abl-homologous exon. These data suggest a role for c-abl in CML, a theory supported by the presence of an abnormally sized abl messenger RNA and protein in the CML cell line K562. The region involved in the translocation on chromosome 22 has also been identified: all Ph'-positive patients examined to date have a breakpoint within a 5.8-kb region, for which we have proposed the name 'breakpoint cluster region' (bcr). To determine whether bcr contains protein encoding regions, probes from bcr were tested for their ability to hybridize to complementary DNA sequences. A 0.6-kb HindIII/BamHI bcr restriction enzyme fragment proved suitable for isolating several cDNA clones from a human fibroblast cDNA library. Using bcr cDNA sequences, we obtained data strongly suggesting the presence of a chimaeric bcr/abl mRNA in the leukaemic cells of Ph' positive CML patients. The recent isolation of cDNA clones containing bcr and abl sequences confirms this finding. Because the bcr part of the chimaeric mRNA could be required to induce the transforming activity of the human c-abl oncogene, we have now initiated studies to characterize the normal 'bcr gene' and to determine the effect of a translocation within its coding domain. We demonstrate that as a result of the Ph' translocation, a variable number of bcr exons are included in the chimaeric bcr/abl mRNA. The bcr gene sequences in this mRNA could be responsible for the transition of the abl cellular proto-oncogene into an oncogene. PMID- 2989704 TI - Laying the groundwork for neutralizing AIDS-linked virus. PMID- 2989705 TI - Is the AIDS virus recombinant? PMID- 2989706 TI - Neutralization of human T-lymphotropic virus type III by sera of AIDS and AIDS risk patients. AB - Human T-lymphotropic virus type III (LAV, HTLV-III) is aetiologically linked to acquired immune deficiency syndrome (AIDS) and persistent general lymphadenopathy (PGL). Specific radioimmunoassays (RIA), enzyme-linked assays, immunofluorescence assays (IFA) and immunoblotting techniques are being used widely to detect serum antibodies to HTLV-III in infected patients and in those at risk of infection. However, these assays do not functionally identify those antibodies that neutralize the infectivity of the virus. We have used three methods of titrating serum neutralizing factors: inhibition of syncytium induction, neutralization of envelope pseudotypes of vesicular stomatitis virus (VSV) and reduction of infectivity of HTLV-III for a cell line permissive to virus replication. We report here that sera from subjects in various disease categories possess only low-level neutralizing activity, even when antibodies to viral membrane antigens are present in high titre. Envelope pseudotypes prepared from four HTLV-III isolates made in three different countries are equally sensitive to neutralization by positive sera, including sera from patients yielding two of the virus isolates. PMID- 2989707 TI - HTLV-III-neutralizing antibodies in patients with AIDS and AIDS-related complex. AB - The isolation of the human T-cell leukaemia (lymphotropic) virus type III (HTLV III or lymphadenopathy-associated virus) from cells of many patients with acquired immune deficiency syndrome (AIDS) presented the first evidence that the virus was the aetiological agent of the disease. Subsequent seroepidemiological studies have shown the presence of HTLV-III-specific antibodies in the serum of most patients with AIDS and AIDS-related complex (ARC), and in the serum of many individuals at risk for AIDS. Despite these extensive studies, there are no reports of protective effects of HTLV-III antibodies. In contrast, neutralizing antibodies specific for HTLV-I and -II have been identified previously. Therefore, we investigated whether HTLV-III-exposed individuals possess antibody activities capable of inhibiting viral infection. Here, we report that natural antibodies capable of neutralizing HTLV-III infection of H9 cells were detected in most adults AIDS and ARC patients but in no normal healthy heterosexual controls. Geometric mean antibody titres in ARC patients were double those in AIDS patients, and were even higher in two antibody-positive healthy homosexuals. This suggests that virus neutralizing antibodies may exert an in vivo protective effect. The presence of these antibodies indicates an immunological response to HTLV-III which potentially may be manipulated for therapeutic advantage. The methodology used here will be useful in monitoring future vaccine approaches. PMID- 2989708 TI - Individual-specific 'fingerprints' of human DNA. AB - Simple tandem-repetitive regions of DNA (or 'minisatellites') which are dispersed in the human genome frequently show substantial length polymorphism arising from unequal exchanges which alter the number of short tandem repeats in a minisatellite. We have shown previously that the repeat elements in a subset of human minisatellites share a common 10-15-base-pair (bp) 'core' sequence which might act as a recombination signal in the generation of these hypervariable regions. A hybridization probe consisting of the core repeated in tandem can detect many highly polymorphic minisatellites simultaneously to provide a set of genetic markers of general use in human linkage analysis. We now show that other variant (core)n probes can detect additional sets of hypervariable minisatellites to produce somatically stable DNA 'fingerprints' which are completely specific to an individual (or to his or her identical twin) and can be applied directly to problems of human identification, including parenthood testing. PMID- 2989709 TI - DNA restriction fragments associated with alpha 1-antitrypsin indicate a single origin for deficiency allele PI Z. AB - The alpha 1-protease inhibitor, or alpha-antitrypsin (AAT), a major plasma inhibitor of leukocyte elastase and bacterial proteases, is encoded at the PI locus on chromosome 14 (14q24.3-q32.1). A deficiency of AAT in individuals homozygous for the PI Z allele occurs in about 1 in 2,000-8,000 caucasians and is associated with an increased risk of early adult onset emphysema and liver disease in childhood. We have now used DNA polymorphisms associated with the AAT gene to investigate the origin of the PI Z allele. Using two genomic probes extending into the 5' and 3' flanking regions, respectively, we have identified eight polymorphic restriction sites. Extensive linkage disequilibrium occurs throughout the probed region with the PI Z allele, but not with normal PI M alleles. The Z allele occurs mainly with one haplotype, indicating a single, relatively recent, origin in caucasians. PMID- 2989710 TI - Spontaneous excision of a large composite transposable element of Drosophila melanogaster. AB - The TE1 family of transposable elements (TEs) of Drosophila consists of unusually large transposons, cytologically visible in larval polytene chromosomes as one or more bands. They are composite elements, as their termini consist of foldback (FB) sequences which are themselves transposable. The location of FB elements at the termini of transposable elements suggests that these sequences have a direct role in the genetic instability of TEs. To investigate the structural and phenotypic consequence of TE excision, we have cloned genomic DNA required for the expression of the no-ocelli (noc) gene of Drosophila; this gene has been mutated by the insertion of TE146, a member of the TE1 family carrying six polytene chromosome bands including functional copies of the white (w+) and roughest (rst+) genes. As reported here, our experiments indicate that the spontaneous excision of TE146, which results in the loss of the w+ and rst+ markers, can occur either as a single-step event or following a partial internal deletion. In either case, the end product is an imprecise excision in which a residual portion of the element, varying in size from 3 to 10 kilobases (kb), is left at the insertion site. These residual sequences share homology with the FB family. Furthermore, despite their imprecise nature, all these spontaneous excisions restore a wild-type noc+ phenotype. PMID- 2989711 TI - Local degradation of fibronectin at sites of expression of the transforming gene product pp60src. AB - Local degradation of extracellular fibronectin, a major extracellular adhesive protein, is believed to play an important part in the migration of cells through the extracellular matrix during tumour invasion, morphogenetic movement and trophoblast implantation. Fibronectin is lost from the cell surface after transformation with Rous sarcoma virus (RSV). By using fluorescent and radiolabelled probes covalently coupled to the surface of substrata, we have recently identified a proteolytic activity that is expressed in RSV-transformed cells and is involved in the local degradation of fibronectin at cell-substratum contact sites. Here, we extend the relevance of these findings and gain some insight into the cellular functions of pp60src, the transforming gene product of RSV. We show that newly expressed viral pp60src is localized at the cytoplasmic surface of the cell membrane, corresponding to the cell contact sites where degradation of extracellular fibronectin occurs. PMID- 2989712 TI - The class I major histocompatibility antigen gene activated in a line of SV40 transformed mouse cells is H-2Dd, not Qa/Tla. AB - We have previously described several complementary DNA clones isolated because they correspond to messenger RNAs present at higher levels in the simian virus 40 (SV40)-transformed BALB/c 3T3 cell line SV3T3 Cl38 than in the normal, parental BALB/c 3T3 line. One of these clones, pAG64, hybridizes to RNAs which, while present in BALB/c 3T3 cells, are 10-20-fold more abundant in SV3T3 Cl38 and are found at high levels in a wide variety of transformed cell lines. Nucleotide sequence analysis showed that pAG64 encodes a class I antigen of the major histocompatibility complex. To ascertain the identity of pAG64, we compared its sequence with the available sequences of d haplotype class I antigen genes [K locus, L locus, D locus and the Qa gene defined by genomic clone 27.1] and found that it showed multiple clustered differences from each of these sequences. We therefore concluded that it was not derived from the H-2Kd, H-2Ld or H-2Dd genes and thus must correspond to one of the other class I antigen genes, namely those of the Qa/Tla complex, although it was clearly not the Qa gene defined by the genomic clone 27.1. We now report subsequent findings which indicate that pAG64 in fact corresponds to the H-2Dd gene and not to a Qa/Tla gene. PMID- 2989713 TI - Amplification of inter-Alu extrachromosomal DNA during cellular ageing: retraction and explanation. AB - We reported previously the age-dependent appearance of extrachromosomal circular DNA bands hybridizing to a human DNA fragment ('inter-Alu'), isolated from a genomic cluster of AluI repeats. Such bands appeared or increased at late passage in four out of six human fibroblast strains (six out of nine cell expansions); moreover, all DNAs (9/9) obtained from peripheral lymphocytes of aged donors, but none (0/8) from young donors, revealed a non-genomic inter-Alu band at congruent to 4.8 kilobases (kb). Subsequent data extended these numbers to 16/24 aged donors compared with 0/18 young donors. These results were interpreted as evidence of age-dependent DNA rearrangement in normal human cells. We now report that the 'extra' bands were of microbial origin, although clearly occurring in an age-dependent manner. PMID- 2989714 TI - 6-Hydroxydopamine pretreatment effects on alpha- and beta-adrenergic receptor adaptation to clorgyline. AB - The effects of 6-hydroxydopamine (6-OHDA) lesions on brain adrenergic receptor adaptation to 21 days of treatment with the selective monoamine oxidase type A (MAO-A) inhibitor clorgyline were studied in rats. 6-OHDA pretreatment effectively blocked the decrease in alpha 1-, alpha 2- and beta-adrenergic receptor densities observed in response to clorgyline treatment. In saline treated rats, 6-OHDA reduced norepinephrine (NE) to 8% of control levels, modestly reduced dopamine (DA) to 67% of controls, but did not affect serotonin (5HT) levels in the cortex, Clorgyline administration to shams increased NE and 5HT to 239% and 160% of their respective control levels, but did not effect DA levels. 6-OHDA lesions attenuated clorgyline's effect on cortical NE levels but not 5HT. The results suggest that beta-adrenergic receptor adaptation to MAOI's as with tricyclic antidepressants is a response to an increase in catecholamine receptor occupancy, and that a similar molecular mechanism is responsible for the observed clorgyline induced changes in alpha-adrenergic receptors. PMID- 2989715 TI - The sympathetic axons innervating the sinus node of the rabbit possess presynaptic opioid kappa- but not mu- or delta-receptors. AB - The postganglionic sympathetic nerves of rabbit isolated hearts were stimulated with pulses delivered at 5 Hz and train durations of 1-5 s. Ethylketocyclazocine 0.01-1 mumol/l and fentanyl 1 and 10 mumol/l but not morphine 1 and 10 mumol/l, Met-enkephalin 1 and 4 mumol/l or D-Ala2, D-Leu5-enkephalin 0.5 and 5 mumol/l diminished the stimulation-evoked increase in heart rate. The effect of ethylketocyclazocine 0.1 mumol/l was antagonized by naloxone 1 and 10 mumol/l. In contrast, the effect of fentanyl was not changed by naloxone 10 mumol/l. Ethylketocyclazocine 0.03 and 1 mumol/l did not reduce the tachycardia elicited by exogenous noradrenaline. The results suggest that, under in vitro conditions, only presynaptic opioid kappa- but not mu- or delta-receptors inhibit the release of noradrenaline from the sympathetic neurones innervating the sinus node. PMID- 2989717 TI - Chemiluminescence and superoxide anions generated by phagocytes in uraemia. PMID- 2989716 TI - Tubular disorders of acid-base and phosphate metabolism. PMID- 2989718 TI - Investigations on myelinogenesis in vitro: a study of the critical period at which thyroid hormone exerts its maximum regulatory effect on the developmental expression of two myelin associated markers in cultured brain cells from embryonic mice. AB - Cultures of cells dissociated from embryonic mouse brain were used to assess the period in which thyroid hormone exerts its maximum influence on the regulation of the expression of two myelin associated metabolites, sulfolipids and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP-ase). Cultures were grown for a specified number of days on a medium containing normal calf serum and then a portion were switched to a medium containing hypothyroid calf serum for 2 days. One half of these cultures were then supplemented with 50 nM triiodothyronine and growth was continued in all cultures for 3 more days. The cells were then assayed for CNP ase activity and for their ability to incorporate 35SO4 into sulfolipids. Studies with both myelin markers showed that in the earlier culture ages of 5, 8, and 11 days, thyroid hormone was able to fully restore the activities when added to cultures grown on hypothyroid calf-serum. In contrast, in the intermediate age range (15, 19, and 22 days) the restoration was partial, while in the higher ages, there was practically negligible restoration with T3. Since the culture system eliminates the possibility of a blood brain barrier and drastically decreases the complicity of other hormones, the lack of a myelinogenic response to thyroid hormone after a certain age must be attributed to the loss of sensitivity of the oligodendroglia to T3 possibly through genetic programming. PMID- 2989719 TI - Characterization of rat brain opioid receptors by [Tyr-3,5-3H]1, D-Ala2, Leu5 enkephalin binding. AB - [Tyr-3,5-3H]1, D-Ala2, Leu5-enkephalin [( 3H]DALA) was used for labeling the opioid receptors of rat brain plasma membranes. The labeled ligand was prepared from [Tyr-3,5-diiodo]1, D-Ala2, Leu5-enkephalin by catalytic reductive dehalogenation in the presence of Pd catalyst. The resulting [Tyr-3,5-3H]1, D Ala2, Leu5-enkephalin had a specific activity of 37.3 Ci/mmol. In the binding experiments steady-state level was reached at 24 degrees C within 45 min. The pseudo first order association rate constant was 0.1 min-1. The dissociation of the receptor-ligand complex was biphasic with k-1-s of 0.009 and 0.025 min-1. The existence of two binding sites was proved by equilibrium studies. The high affinity site showed a KD = 0.7 nM and Bmax = 60 fmol/mg protein; the low affinity site had a KD = 5 nM and Bmax = 160 fmol/mg protein. A series of opioid peptides inhibited [3H]DALA binding more efficiently than morphine-like drugs suggesting that labeled ligand binds preferentially to the delta subtype of opioid receptors. Modification of the original peptides either at the C or N terminal ends of the molecules resulted in a decrease in their affinity. PMID- 2989720 TI - [Ultrastructure of capillary permeability in human brain tumors. Part 1: Gliomas associated with cerebral edema (low density area)]. AB - In order to elucidate pathogenesis of perifocal edema in the human brain tumors, we observed the alteration of capillary permeability between the glioblastomas with remarkable edema (4 cases) and astrocytoma with slight edema (3 cases). Specimens were studied by conventional ultrathin section and freeze-fracture replica technique. In ultrathin sections of capillaries in glioblastomas, some of these cell junctions were tortuous, elongated, in fact, open. Other capillary abnormalities included endothelial hyperplasia with extensive vesicular formation, surface infolding of endothelial cells, irregularity of the basal lamina and the presence of a large collagen filled extracellular space. In freeze fracture replicas of capillary endothelium, pinocytotic vesicles markedly increased and were an average fo 52 per micron. Tight junction in one area was seen as network of 6 strands composed of about 100A particles, but in the other areas as one or two strands. In ultrathin sections of astrocytoma, yet there were blood vessels appeared relatively normal. In freeze-fracture replicas, pinocytotic vesicles markedly increased and were an average of 34 per micron. Tight junction was seen as network of 7 strands. We concluded that fewer strands of the tight junction play an important role in increasing the permeability in the vessels of glioblastomas with severe perifocal edema, in addition to increasing the pinocytotic vesicles. PMID- 2989721 TI - [Case of intracranial and spinal dissemination of primary spinal glioma]. AB - A case of glioblastoma multiforme of spinal cord followed by intracranial dissemination was reported. A 20-year-old man was admitted on Sep. 22, 1981, with motor disturbance of both legs and loss of urinary control. Neurological examination showed spastic paraplegia with bilateral Babinski signs. Abdominal and cremaster reflexes were absent. There was no abnormalities in cranial nerves and upper limbs. Cerebral CT was normal at that time. Myelography showed complete block at the level of 8th thoracic vertebra. Spinal angiography revealed abnormal vascularity at the level of Th7 fed via anterior spinal artery. Intramedullary tumor was partially removed through 5th-8th thoracic laminectomy. Total 5600 rads of irradiation was administered after the operation. The postoperative course was deteriorating. Two months after the operation paresis of both upper extremities had been developed. State of consciousness had been disturbed due to increased intracranial pressure which was subsided for a time while by ventriculoperitoneal shunt. Repeated CT showed high density spots scattered in basal cisterns spreading to all ventricular systems. He expired Feb. 21, 1982. Autopsy revealed that the whole spinal cord was covered by tumor tissue. Basal cisterns were filled with tumor tissue. The tumor was infiltrated subependymally into ventricular systems with intramedullary invasions in some places. Histological diagnosis was glioblastoma multiforme. PMID- 2989722 TI - Corticotropin-releasing factor receptors in human pituitary gland: autoradiographic localization. AB - Corticotropin-releasing factor (CRF) receptors were localized in the human pituitary gland by an in vitro labeling light microscopic autoradiographic technique using an 125I-labeled analog of ovine CRF substituted with norleucine and tyrosine at amino acid residues 21 and 32, respectively. Specific binding sites for CRF were observed in the anterior lobe while no specific binding sites for CRF were present in the posterior pituitary lobe. The CRF binding sites in the anterior lobe occurred in clusters which predominated in the anteromedial portion of the lobe. This pattern of localization of CRF binding sites resembles previously reported distributions of corticotrophs in anterior pituitary. These data support the physiological role of endogenous CRF in regulating hormone secretion from the anterior lobe of the human pituitary. PMID- 2989724 TI - Responsiveness of immature versus adult male rat hypothalami to dibutyryl cyclic AMP- and forskolin-induced LHRH release in vitro. AB - In the present study, we have investigated the effects of intermittent dibutyryl cyclic AMP (dbcAMP, 5 X 10(-8) M), butyrate (5 X 10(-8) M) and forskolin (10(-4) M) on immunoreactive luteinizing hormone-releasing hormone (LHRH) release from superfused hypothalamic fragments from intact male rats of age 25, 30, 45, or 60 75 day (adult). The results indicate that at 25 days of age, male rat hypothalami were most responsive to cyclic AMP (162% of preinfusion basal LHRH release); by 30 days of age, dbcAMP also elicited increased LHRH release (120% of basal). By 45 days of age, the dbcAMP effect on in vitro LHRH release was slightly inhibitory (78% of basal); however, by adulthood, the effect of this cyclic nucleotide on LHRH release was minimal (92% of basal). Butyrate also induced age dependent modifications in in vitro LHRH release from male rat hypothalami, with slight increases following butyrate delivery at 25 days of age (115%), slight decreases at 30 (82%) and 45 days of age (68%), and little change in adulthood (94%). This latter finding emphasizes the importance of using butyrate as a control for butyryl derivatives of cyclic AMP, which are known to liberate butyric acid as a product of hydrolysis of parent compounds. To address the possibility that the lack of effectiveness of dbcAMP in the older animal preparations was solely due to an increasing sensitivity of male rat medio-basal hypothalamus fragments to butyrate, we examined the effect of forskolin (10(-4) M), an adenylate cyclase stimulator, on LHRH release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989723 TI - Apparent lack of involvement of cAMP as a mediator of LHRH stimulation of nuclear estrogen receptor activity in the rat anterior pituitary. AB - We have previously shown that incubation of rat pituitary cells in the presence of LHRH results in specific enhancement of nuclear estrogen receptor (ER) binding which cannot be accounted for by simple intracellular translocation of cytoplasmic receptor. In the present study, the role of cAMP in this response has been examined. Suspended pituitary cells from adult ovariectomized rats primed with estradiol were incubated with varying concentrations of LHRH or a highly active LHRH analog (LHRH-A) for 30 min at 37 degrees C, and levels of cAMP were determined. Total cAMP levels changed only in response to a concentration of 100 pmol/pituitary of either peptide; the stimulation by LHRH was twice that by LHRH A. When the priming dose level of estradiol was reduced from 1.0 to 0.5 micrograms/day, stimulation of cells by 100 pmol of LHRH caused a much greater increase in cAMP levels. Separate incubation of subcellular fractions with a wide dose range of dibutyryl-cAMP (DBcAMP) resulted in a progressive loss of cytosol receptor binding capacity (which was also observed in whole cultured or suspended cells), but no significant concomitant change in nuclear receptor binding; however, whole cells in suspension or culture did show an increase in nuclear ER activity in the presence of 100 nM DBcAMP. This response was qualitatively, but not quantitatively, similar to that elicited by LHRH. When the effects of whole cell incubation with LHRH and LHRH-A on nuclear ER were compared with their effects on total cAMP levels, no correlation was observed; rising levels of cAMP did, however, coincide with falling levels of cytosol ER.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989725 TI - An inborn error of purine metabolism, deafness and neurodevelopmental abnormality. AB - A syndrome of hyperuricemia, sensorineural deafness, mild mental handicap and congenital disequilibrium in a four-year-old boy is probably inherited as a sex linked condition since his mother has sensorineural deafness and similar biochemical abnormalities. There is evidence of a superactive PP-ribose-P synthetase, normal purine salvage enzymes, and severe depletion of nicotinamide adenine dinucleotide and guanine triphosphate in red cells. PMID- 2989726 TI - Metastasis of an occult gastric carcinoma suggesting growth of a prolactinoma during bromocriptine therapy: a case report with a review of the literature. AB - The treatment of a slowly growing invasive prolactinoma with bromocriptine for 8 months resulted in a substantial decrease in plasma prolactin levels despite rapid suprasellar tumor expansion. On exploration, this uncommon observation could be attributed to hematogenous metastasis from an occult gastric adenocarcinoma to the pituitary tumor. Apart from infiltration of neighboring parts of the hypothalamus, autopsy revealed no other hematogenous metastases. This extraordinary type of neoplasm-to-neoplasm metastasis was not shown by computed tomography. This possibility should be considered whenever progressive growth of a pituitary mass is accompanied by a decrease in hormonal overproduction. PMID- 2989727 TI - Nuclear magnetic resonance imaging (NMR), (MRI), of brain stem tumours. AB - The NMR scans of twenty-six patients who were referred with the clinically suspected or histologically verified diagnoses of brain stem tumour were reviewed. Twenty-one patients (eleven children and ten adults) had scans that were positive for tumour. Nine of these had histological confirmation. Eleven were irradiated without histology on the basis of the clinical diagnosis. The remaining patient was treated expectantly. The other five patients (two children and three adults) had CT and NMR scans that were negative for tumours. Clinical follow-up of these five cases for 9 to 29 months disclosed no further evidence of tumour. All but one tumour had evidence of an increased T1 and T2. Mass effects were seen in all but one case (not the same as the first exception). Multiplanar imaging was useful in determining intra- or extra-axial location as well as tumour extent. Intravenous Gadolinium-DTPA (a paramagnetic contrast agent) showed tumour enhancement in the two patients in whom it was used. NMR showed more extensive abnormality than CT in nineteen patients and similar abnormality to CT in two patients. CT demonstrated calcification better than NMR in the three cases in which it was observed. PMID- 2989728 TI - MRI (NMR) in the diagnosis of brain-stem tumors. AB - Patients with a brain-stem tumor were studied with NMR. The full extent of the lesion as well as its relationship with the adjacent structures was clearly demonstrated in all cases. Although NMR is, in many aspects, superior to CT and angiography, these examinations remain useful complementary methods. PMID- 2989729 TI - Behavioural effects in rats of unilateral and bilateral injections of opiate receptor agonists into the globus pallidus. AB - Unilateral injections of a putative kappa-opiate receptor agonist, ethylketocyclazocine, into the globus pallidus of rats caused dose-dependent ipsiversive circling which was inhibited by prior systemic administration of the opiate receptor antagonist naloxone. Neither a putative delta-opiate receptor agonist ( [D-Ala2, D-Leu5]enkephalin) nor a putative mu-opiate receptor agonist (Tyr-D-Ala-Gly-MePhe-Met(0)-ol) induced circling behaviour after unilateral intrapallidal injection. Bilateral intrapallidal injection of the delta-opiate receptor agonist or the mu-opiate receptor agonist caused an increase in locomotor activity which in both cases was reduced by systemic administration of naloxone. A specific delta-receptor antagonist, (N,N-bisalyl-Tyr-Gly-Gly-psi (CH2S)-Phe-Leu-OH) had no effect on the increase in locomotor activity caused by the delta-opiate receptor agonist. Bilateral intrapallidal injection of the kappa opiate receptor agonist had no effect on locomotor activity. It is suggested that different opiate receptor subtypes within the globus pallidus differentially mediate circling and locomotor behaviour. PMID- 2989730 TI - A new complication of AIDS: thoracic myelitis caused by herpes simplex virus. AB - Progressive thoracic myelopathy occurred in a patient with AIDS. Concurrent opportunistic infections included disseminated systemic cytomegalovirus, aspergillosis, and cutaneous herpes simplex virus (HSV). At autopsy, immune stains indicated that the myelopathy was caused by HSV type 2 infection of the spinal cord. PMID- 2989731 TI - [Splenectomy in the radical surgical treatment of ovarian carcinoma]. PMID- 2989732 TI - gamma-Aminobutyric acidA (GABAA) receptors modulate [3H]GABA release from isolated neuronal growth cones in the rat. AB - Potassium-induced release of gamma-[3H]aminobutyric acid [( 3H]GABA) from a growth cone-enriched fraction isolated from neonatal rat forebrain was inhibited by the GABA mimetic muscimol in a dose-dependent manner (IC50 15 nM). The GABA antagonist bicuculline completely reversed the effect of muscimol. Bicuculline alone slightly potentiated the K+-induced release of [3H]GABA. Baclofen, a proposed selective agonist for a bicuculline-insensitive GABAB receptor, was found to cause only a slight reduction in the K+-induced release of [3H]GABA. These results are compatible with the presence of a negative feedback mechanism mediated by GABAA receptors for controlling [3H]GABA release from growth cones of the developing rat forebrain. PMID- 2989733 TI - Evidence for a kappa-opioid receptor on pituitary astrocytes: an autoradiographic study. AB - Specific binding of the opioid ligand [3H]etorphine was localized in cryostat sections of the rat pituitary. The binding was concentrated over the pars nervosa. Competition with partially selective unlabelled displacers indicated that the greater part of this binding was of the kappa-subtype, sensitive to inhibition by dynorphin and bremazocine. The kappa nature of the binding was confirmed using the prototype kappa ligand [3H]ethylketocyclazocine in the presence of excess unlabelled mu and delta opioid receptor ligands. Sections of the pituitary stalk caused a loss of the hypothalamic projection to the pars nervosa and a marked gliosis. Concurrent with these changes there was a significant increase in the density of kappa opioid binding. These observations suggest that the majority of opioid receptors in the pars nervosa are of the kappa subtype and are located on the pituicytes. PMID- 2989734 TI - gamma-Aminobutyric acid partly mediates the pentobarbitone depression of synaptic excitation in the guinea-pig olfactory cortex in vitro. AB - Pentobarbitone depresses synaptic excitation in the guinea-pig olfactory cortex slice in vitro. A study has been made to elucidate the possible role of gamma aminobutyric acid (GABA) in this depression by testing pentobarbitone in the presence of high concentrations of the GABA blockers, i.e. picrotoxin or bicuculline. These blockers reduced the action of pentobarbitone; the dose depression curve for pentobarbitone was shifted to the right by a factor of 2.3. It is concluded that pentobarbitone has a bimodal action, one action via GABA and another unrelated to GABA or Cl- conductances. PMID- 2989735 TI - Autoradiographic localization of binding sites for the arginine-vasopressin (VP) metabolite, VP4-9, in rat brain. AB - A discrete neuro-anatomical pattern of binding sites was observed for a principal metabolite of arginine-vasopressin (VP4-9) after incubation of tissue sections with [35S]VP4-9 and autoradiography. [35S]VP4-9-labeled binding sites were highly concentrated in the pineal gland, the nucleus tractus solitarii (nts), the arcuate nucleus region (an) and the organum vasculosum lamina terminalis (ovlt). The distribution of these sites is distinctly different from the putative VP and oxytocin receptor systems in rat brain. It is possible that the VP4-9 binding sites are also involved in memory processes and/or that the VP metabolite exerts additional effects on the brain. PMID- 2989736 TI - Barbiturate-enhanced paired-pulse depression in neonatal rats. AB - Paired-pulse depression of the population spike in the hippocampal formation of barbiturate anaesthetized mature rats is increased compared to that in urethane unanaesthetized rats. This barbiturate-induced increase in depression is greatly enhanced in immature animals, which are also known to be behaviorally more sensitive to barbiturates. These results suggest an increased sensitivity of the recurrent inhibitory (possible GABAergic) system to barbiturates in the immature central nervous system. PMID- 2989738 TI - Differential regulation of corticotropin-releasing factor receptors in anterior and intermediate lobes of pituitary and in brain following adrenalectomy in rats. AB - The effects of adrenalectomy on corticotropin-releasing factor (CRF) receptors in anterior and intermediate lobes of rat pituitary and in forebrain were examined using in vitro autoradiography with the radioiodinated analogue of ovine CRF, Nle21, [125I]Tyr32-CRF. The concentration of CRF receptors in the anterior pituitary was significantly reduced at 4 days and remained decreased at 9 weeks after adrenalectomy. In contrast, adrenalectomy did not alter CRF receptors in the intermediate lobe or in a variety of forebrain regions. The adrenalectomy induced change in CRF receptors in the anterior pituitary was completely reversed by glucocorticoid replacement with dexamethasone. These data indicate that endogenous CRF is capable of modulating its receptor density in the anterior pituitary and suggest that different sources of CRF or other factors may be important in regulating intermediate lobe hormone secretion and neuronal activity in brain. PMID- 2989737 TI - Vasoactive intestinal peptide receptor localization in rat forebrain by autoradiography. AB - Vasoactive intestinal peptide (VIP) receptors were localized in rat forebrain by in vitro labeling light microscopic autoradiography with 125I-labeled VIP. Binding sites for VIP were found in discrete areas of rat forebrain including lamina I of the neocortex and pyriform cortex, caudate-putamen, the hippocampus and molecular layer of the dentate gyrus, basolateral nucleus of the amygdala, several thalamic nuclei and the magnocellular paraventricular and supraoptic nuclei of the hypothalamus. These results are consistent with earlier findings on the immunohistochemical distribution and proposed sites of action of VIP, and reinforce the concept that endogenous VIP may function as a neuromodulator in brain. PMID- 2989739 TI - Morphological evidence that serotonin-immunoreactive neurons in the nucleus dorsomedialis hypothalami could be under catecholaminergic influence. AB - In the nucleus dorsomedialis hypothalami (NDM), it has so far been possible to visualize serotonin (5-HT) neurons by 5-HT immunohistochemistry only after inhibition of the monoamine oxidase and loading with tryptophan. We report here that the intraventricular administration of dopamine, noradrenaline (NA) and adrenaline as well as isoproterenol and, to a lesser extent, apomorphine, similarly permitted 5-HT immunostaining of these neurons in pargyline-pretreated rats, and that additional pretreatment with propranolol prevented the NA-induced effect. Hence, it is proposed that the 5-HT content in the presumed serotoninergic neurons of the NDM could be under dopaminergic and/or beta adrenergic receptor-mediated catecholaminergic influence. PMID- 2989740 TI - gamma-Aminobutyric acidB receptors in the rat brain: quantitative autoradiographic localization using [3H](-)-baclofen. AB - The light microscopic localization of gamma-aminobutyric acidB (GABAB) receptors in the rat brain has been accomplished using in vitro autoradiography with [3H]baclofen as a ligand. Initial biochemical studies indicated this compound specifically bound to slide-mounted tissue sections with high affinity in a saturable and readily reversible manner. Autoradiograms generated by labeled tissue sections demonstrated high grain densities over structures such as the interpeduncular nucleus, molecular layer of the cerebellum, superior colliculus, several thalamic nuclei and the superficial laminae of the cortex. The majority of the GABAB sites were found to exist in regions of the brain previously reported to have significant densities of GABA sites present. A few areas were noted, however, where the distribution of GABAA and GABAB sites did not overlap. PMID- 2989741 TI - Nursing alert. Understanding the spectrum of cerebral stimulants (continuing education). PMID- 2989742 TI - NursingLife's guide to hidden hazards on the job. Part I: Infectious diseases. PMID- 2989743 TI - Neuroendocrine regulation of seasonal reproductive activity in the male golden hamster. AB - Golden (Syrian) hamsters are seasonal breeders. Under natural photoperiodic conditions, their reproductive systems are functional during spring and summer and atrophic during the fall and winter. This reproductive cycle can be duplicated in the laboratory by exposing the animals to artificially-created photoperiods. The endocrine correlates of photoperiod-induced changes in reproductive activity of the male hamster are fairly well characterized, but the neural control of seasonal reproductive activity has not been as extensively studied. Recent studies indicate that short day (less than 12.5 hr light/day) exposure leads to complex changes in central neurotransmitter metabolism, as well as neurotransmitter and hormonal receptor content, which, in turn, are reversed by exposure to long days or during the period of spontaneous testicular recrudescence. Many of these endocrine and neuroendocrine changes are dependent on the presence of the pineal gland, but photoperiod-induced changes in neurotransmitter metabolism have also been described in pinealectomized hamsters. Further studies of the neuroendocrine transduction of photoperiodic signals will not only provide a better understanding of seasonal reproductive and metabolic activities, but will increase our basic understanding of the neural control of the endocrine system. PMID- 2989744 TI - Chronic acidosis with metabolic bone disease. AB - The case is reported of a woman in whom a chronic hyperchloraemic acidosis and osteomalacia developed follow implantation of her ureters into the rectum. For 14 years she took regular sodium bicarbonate, but a little over four years after discontinuing it she developed clinical, biochemical, radiological and radionuclide evidence of osteomalacia. The latter improved rapidly when sodium bicarbonate was recommenced. This clinical situation is one of the few instances in man where a chronic metabolic acidosis in the context of normal or only mildly impaired renal function can result in osteomalacia. PMID- 2989745 TI - Malignant fibrous histiocytoma of the vulva. AB - Primary sarcomas of the vulva are rare neoplasms; malignant fibrous histiocytoma is a sarcoma that usually occurs in the extremities of elderly patients, but it also has been seen in a number of other sites. Presented is the second case report of malignant fibrous histiocytoma arising in the vulva. The diagnosis and management of vulvar sarcomas are reviewed. PMID- 2989747 TI - [Models of after-care programs for selected rheumatic diseases following rehabilitation measures]. PMID- 2989746 TI - Effects of estrogen therapy on vaginal physiology during menopause. AB - Vaginal physiology was evaluated in 23 postmenopausal women before estrogen replacement therapy and at 1, 3, 6, 12, 18, and 24 months while receiving conjugated equine estrogens (Premarin). Reversal of hormonal levels (17 beta estradiol, gonadotropins) and vaginal cytology occurred within one month. Vaginal pH levels significantly decreased from a baseline mean of 5.2 to a level of 4.2 at 24 months (P less than .05). Women who were sexually active showed a greater decline in pH levels than did women who were sexually inactive. Maximum increases in amount of vaginal fluid and potassium levels were observed after three months of therapy. Vaginal blood flow and vaginal electropotential difference were significantly increased over baseline values at one month and again at 12 months (P less than .05) with a slow progressive improvement continuing throughout 24 months of estrogen replacement therapy. This study provides documented laboratory evidence to suggest that restoration of vaginal tissue function requires 18 to 24 months and explains why dyspareunia may persist in the early months of replacement therapy despite hormonal and cytologic return to premenopausal values. PMID- 2989748 TI - Clinical observations concerning choroidal folds. AB - This paper deals with 6 patients who suffer from choroidal folds. Special emphasis will be laid on several aspects of differential diagnostics. In our 6 cases discussed here, we found a retrobulbar tumor in 3 patients, hypotony in 1, and a marked hyperopia in another as causes. In the 6th patient no pathological condition was found, and, therefore, we characterized the choroidal folds in this case as idiopathic. PMID- 2989750 TI - Salivary epithelial-myoepithelial carcinomas of intercalated ducts: a clinical, electron microscopic, and immunocytochemical study. AB - The salivary epithelial-myoepithelial carcinoma of intercalated duct origin is a distinctive, biologically low-grade carcinoma with a predilection for the parotid gland. Nine examples from the M.D. Anderson Hospital in Houston, Texas, bring the number of published cases to 33. Immunocytochemical (S-100 protein, myosin, and keratin) and electron-optic studies strongly support an active myoepithelial cell participation in the histogenesis of these carcinomas. PMID- 2989749 TI - Juvenile nasopharyngeal angiofibroma: an unusual presentation in the oral cavity. AB - The juvenile nasopharyngeal angiofibroma is a vascular neoplasm that is rarely encountered in the oral cavity. An unusual case of a juvenile nasopharyngeal angiofibroma that presented initially as an asymptomatic palatal mass is reported. The clinical features, diagnosis, and management of this lesion are discussed. PMID- 2989751 TI - Dentinogenesis imperfecta type III with enamel and cementum defects. AB - Ground sections and scanning electron microscopic studies of primary teeth from a 2-year-old child whose teeth showed absence of enamel and large pulp chambers are examined. The findings suggest a diagnosis of odontodysplasia in a case classified as dentinogenesis imperfecta type III. PMID- 2989752 TI - Epithelial-myoepithelial carcinoma: an immunocytochemical study. AB - An immunocytochemical study of epithelial-myoepithelial carcinoma, using antibodies to smooth muscle myosin, keratins, and type IV collagen, is presented. This rare tumor of salivary gland is composed of tubules and ductal structures, comprising an outer layer of clear cells that show myoepithelial differentiation and an inner layer of eosinophilic cells that show ductal differentiation. The findings in this study correlate well with previous ultrastructural descriptions of this tumor. PMID- 2989753 TI - Detection of human papovavirus antigen in oral papillary lesions. PMID- 2989754 TI - The "pedicle flap": a technique for complete excision of benign salivary gland tumors of the palate. AB - Morphology and growth pattern of the pleomorphic adenoma are discussed. The relationship between these factors and recurrence is examined. A new technique for excision is also described. This technique is particularly useful for those tumors that extend into the soft palate. PMID- 2989755 TI - Characterization of cells in salivary gland lesions by immunohistochemical identification of carcinoembryonic antigens. AB - Immunohistochemical demonstration of carcinoembryonic antigen (CEA) was reported in normal salivary glands and in pathologic lesions. Staining patterns of CEA and nonspecific cross-reacting antigen-absorbed CEA (NCAa-CEA) were compared. Normal salivary glands disclosed positive staining by CEA on border and luminal sides of acinar cells and occasionally in components secreted into ductal spaces with both antigens used. Chronic obstructive lesions displayed an intense CEA staining in ductlike structures and in material secreted into their lumina in the two antigens used. Pleomorphic adenoma exhibited varying intensities of CEA in neoplastic epithelial cells of ductal structures. In contrast, they showed a slight staining reaction to NCAa-CEA. Squamous-cell carcinomas showed a strong CEA reaction, whereas they showed no reaction or a trace reaction to NCAa-CEA. Positive staining to CEA in squamous neoplastic lesions was related to nonspecific reacting antigens. PMID- 2989756 TI - Physiologic acoustic basis of speech perception. AB - In the past, there has been a distinction between extraction of the auditory parameters of speech and a specialized phonetic speech processor. It was suggested that for the latter, exclusive specialized processing is required by the dominant hemisphere, while the former was handled by the general auditory system shared by both hemispheres. Present research shows that the peculiarity of speech does not lie in its phonologic information but in the acoustic information which the central auditory system processes. It is, therefore, suggested that specialization for the perceptual process extends to the mechanisms by which the acoustic features of speech are extracted and processed. PMID- 2989757 TI - Lobules in the nipple. Frequency and significance for breast cancer treatment. AB - Subcutaneous mastectomy with nipple preservation has been advocated for the treatment of in situ lobular carcinoma. The rationale for not removing the nipple in these cases is the perception that lobules do not occur in the nipple. This study was undertaken to assess the frequency with which lobules occur in the nipple, exclusive of the areola. A routine vertical section of the nipple was reviewed in each of 101 consecutive mastectomies performed for breast carcinoma. One or more lobules was found in 17 percent of specimens. An additional independent observation of note was the presence of carcinoma in 13 percent of nipples, largely noninvasive. Coexistence of lobules and carcinoma was found in two cases. These data indicate that nipple preservation is not appropriate if mastectomy is performed to remove tissue at risk in the treatment of breast cancer. A procedure which removes the central part of the nipple or nipple reconstruction should be employed. PMID- 2989758 TI - Histologic features of hepatocellular carcinoma and allied disorders. PMID- 2989759 TI - Deranged alpha-adrenergic regulation of growth hormone secretion in poorly controlled diabetes: reversal of the exaggerated response to clonidine after continuous subcutaneous insulin infusion. AB - Elevated plasma growth hormone (GH) and peripheral catecholamine levels are frequently observed in poorly controlled, insulin-dependent diabetes. Since the alpha adrenergic system plays an important role in hypothalamic regulation of GH secretion, we tested the hypothesis that altered central adrenergic activity contributes to the increased GH concentrations in diabetes. Clonidine, an alpha adrenergic agonist, was administered to nine poorly controlled, young diabetic patients (age 12-19 yr) before and after 1 wk of continuous subcutaneous insulin infusion pump therapy. As expected, continuous subcutaneous insulin infusion lowered mean 24-h plasma glucose (from 203 +/- 21 to 112 +/- 7 mg/dl, p less than 0.01) and GH (from 17.7 +/- 2.1 to 9.2 +/- 1.2 ng/ml, p less than 0.01) to values observed in normal controls. In the diabetic patients during conventional treatment, both the peak plasma GH level postclonidine (48.3 +/- 8.7 ng/ml) and the incremental area under the GH response curve (3.23 +/- 0.58 mg X min/ml) were significantly increased above normal control values (25.2 +/- 2.1 ng/ml, p less than 0.05 and 1.63 +/- 0.11 mg X min/ml, p less than 0.0025, respectively). In contrast, the GH response to clonidine was indistinguishable from normal after only 1 wk of intensified insulin treatment. Our findings support the contention that metabolic control of diabetes influences hypothalamic regulation of GH secretion and suggests that such alterations are related, at least in part, to changes in central alpha-adrenergic activity. PMID- 2989761 TI - Mode of inhibitory action of bilirubin on protein kinase C. AB - Ca2+-activated, phospholipid-dependent protein kinase (protein kinase C) is widely distributed in mammalian tissues. Accumulating evidence has revealed that protein kinase C as well as cAMP-dependent protein kinase plays important roles in various cellular functions. The purpose of this study is to examine the effect of bilirubin on protein kinase C and cAMP-dependent protein kinase activity in a cell-free system as a cause of bilirubin toxicity to the central nervous system. Bilirubin inhibited protein kinase C activity in a dose-dependent manner. This effect was markedly diminished by the addition of human serum albumin at a molar ratio of bilirubin to albumin of less than 1.0. Kinetic analysis revealed that bilirubin did not compete with phospholipid, diacylglycerol, or calcium. Bilirubin also inhibited cAMP-dependent protein kinase, but did not compete with cAMP. The inhibitory effect of bilirubin on protein kinase C seems to be irreversible because removal of bilirubin by Sephadex G-25 column chromatography did not restore the protein kinase C activity. Observations reported herein suggest that bilirubin, especially in its free form, induces an irreversible change to the catalytically active site of protein kinase C. PMID- 2989762 TI - Fatal staphylococcal epidermidis infections in very low-birth-weight infants with cytomegalovirus infection. PMID- 2989760 TI - T-lymphocyte subpopulations and function during murine cytomegalovirus infection. AB - To study the effects of cytomegalovirus infection on T-lymphocyte subpopulations, we determined helper (Lyt 1.2) and suppressor (Lyt 2.2) T-lymphocyte subset numbers using monoclonal antibodies and measured lymphocyte responsiveness to mitogen during sublethal murine cytomegalovirus (MCMV) infection of 3-wk-old Balb/c mice. MCMV-infected mice had reduced Lyt 1.2 to Lyt 2.2 T-lymphocyte ratios on days 1, 3, 5, and 9 of infection. Alterations in T-lymphocyte subsets were accompanied by diminished lymphocyte response to concanavalin A. Lymphocyte responsiveness and Lyt 1.2 to Lyt 2.2 ratios were maximally reduced on day 5 of MCMV infection and correlated strongly with peak virus recovery from spleen, bone marrow, and peripheral blood leukocytes. These results indicate that acute MCMV infection of mice causes abnormalities in T-lymphocyte subset ratios and responsiveness to mitogen similar to the abnormalities observed in human cytomegalovirus infections. MCMV infection of mice is a useful model to study the mechanism by which cytomegalovirus infections induce altered T-lymphocyte subpopulations. PMID- 2989763 TI - Coxsackievirus group B antibodies in the ventricular fluid of infants with severe anatomic defects in the central nervous system. AB - Ventricular fluids from four of 28 newborn infants who were initially seen with severe congenital anatomic defects in the central nervous system contained neutralizing antibody to at least one serotype of coxsackieviruses group B. Two of the four infants with anti-coxsackieviruses group B antibody in the ventricular fluid did not have a detectable level of the same antibody(ies) in their serum. The ventricular fluid of one of the infants had immunoglobulin M neutralizing antibody directed against coxsackievirus B6. Of 11 mother-infant pairs that had neutralizing antibody to coxsackieviruses group B in both sera, nearly half had antibodies directed against more than one serotype. These data suggest the possibility of an association between congenital infections with coxsackieviruses group B and rare severe CNS defects. PMID- 2989764 TI - Adenoviral diseases in children: a study of 105 hospital cases. AB - The clinical findings for 105 children hospitalized with adenoviral infection were studied prospectively. In 82 children, the diagnosis was based on the detection of adenovirus antigen in the nasopharyngeal specimens and in 17 children in the feces. In the remaining six patients, findings from nasopharyngeal specimens were negative but a significant increase in CF (complement fixation) titers was detected. The clinical picture of adenoviral infection was characterized by high-grade (mean 39.4 degrees C) and prolonged fever (mean duration 5.4 days). Tonsillitis, otitis, and gastroenteritis were the most common illnesses. In 17% of the patients, no identifiable focus of infection could be demonstrated; nine children with no identifiable focus of infection had febrile convulsions. The WBC count and ESR varied from normal values to values seen in bacterial infections; thus it was difficult to distinguish adenoviral disease from a bacterial disease. Forty-five children were referred to the hospital due to infection unresponsive to antimicrobial therapy. The rapid detection of adenovirus antigen in nasopharyngeal specimens or feces proved to have a great clinical value in the diagnosis of adenoviral infections. PMID- 2989765 TI - Series dead space for inert gases in healthy subjects. AB - In ten normal subjects, series dead space was determined for six intravenously infused inert gases (SF6, ethane, cyclopropane, fluothane, ether, acetone) from their expired and alveolar concentrations. The method for sampling alveolar gas was based on the criterion of identity of mean alveolar and expired gas exchange ratios. Inert gases were analysed chromatographically. Acetone, the most soluble gas, yielded the lowest dead space, the difference to the other gases being about 4.5%. This is probably due to the non-infinite value of the series dead space ventilation-perfusion ratio (VA/Q) which was estimated at about 2,000. The diffusivity, inversely related to the molecular weight, also played a role, the heaviest gas (fluothane) having a greater dead space than the lightest (ethane). The underestimation of the dead space from acetone is expected to be greater in subjects with low tidal volume and high bronchial blood flow, i. e. in some patients with respiratory disease. PMID- 2989766 TI - Possible mechanism of adrenergic and nonadrenergic inhibition in intestinal smooth muscle cells. AB - Nonadrenergic synaptic transmission in circular and longitudinal smooth muscles of caecum preexposed to K-free solution for 4-5 h has been studied by means of sucrose gap technique. In addition, the effects of noradrenaline (NA) and ATP on these muscles were investigated under these conditions. The action of the above substances was accompanied by depolarization and contraction. NA induced a decrease in the membrane resistance. Addition of 0.5 mM Ba2+ to K-free solution intensified the depolarization. 1 mM of Mn2+ blocked depolarization and contraction. Intramural stimulation produced noncholinergic e.j.p.s blocked by TTX. Addition of 0.5 mM Ba2+ increased their amplitude. A reversal potential of both NA-induced depolarization and e.j.p. was in the range of + 10 to + 20 mV. It is supposed that e.j.p.s and depolarization observed in response to ATP and NA action are due to an increase in calcium permeability of the membrane. PMID- 2989768 TI - Transmission of low temperature information in the rat trigeminal system. AB - Recordings have been made from cold-receptive neurons in trigeminal nucleus caudalis to define the lower receptive field temperatures at which the neurons become silent. Most cells were found to become silent at 0 degrees C, but a few were still active at skin temperatures of -5 degrees C. PMID- 2989767 TI - Parallel changes in red blood cell and renal Na-K-ATPase activity in adrenal and electrolyte disorders in the rat. AB - To compare the activity of Na-K-ATPase in the red blood cells (RBCs) and in renal tissue in disorders of Na+ metabolism, the following groups of rats were studied: 1) control, intact rats, 2) adrenalectomized (ADX) rats, 3) intact rats treated with DOCA, 4) ADX DOCA-treated rats, 5) intact salt-loaded rats, 6) ADX salt loaded rats, 7) intact dexamethasone-treated rats (DEXA), and 8) ADX DEXA-treated rats. After adrenalectomy (group 2) serum Na+ decreased and serum K+ increased. Renal Na-K-ATPase in cortex, medulla and papilla of the control group was 44 +/- 2.7 mumol Pi/mg prot/h, 128.2 +/- 5.9 and 44 +/- 3.2 respectively and in group 2 the enzyme activity was 32.5 +/- 2.0 (P less than 0.005), 81.7 +/- 4.5 (P less than 0.001) and 23.6 +/- 1.9 (P less than 0.001) respectively. RBCs Na-K-ATPase of control animals was 2.82 +/- 0.19 mumol Pi/mg prot/h, while in group 2 the activity was 1.43 +/- 0.24 (P less than 0.001). DOCA treatment of ADX rats (group 4) normalized serum electrolytes and Na-K-ATPase activity in the renal cortex and papilla and in the RBCs. In the renal medulla the correction by DOCA was only partial. Salt loading of ADX rats (group 6) normalized serum electrolytes and Na K-ATPase activity in the renal medulla and RBCs. Salt loading of normal rats increased RBC Na-K-ATPase to 3.72 +/- 0.36 (P less than 0.02) and medullary Na-K ATPase to 185.6 +/- 9.8 (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989770 TI - [Application of DSA in the diagnosis of breast cancer]. PMID- 2989769 TI - Descending projections from the ventrolateral medulla and cardiovascular control. AB - Antidromic responses were evoked in neurones in nucleus paragigantocellularis lateralis (PGL) by stimulation of the ipsilateral ventrolateral white matter or the contralateral dorsolateral white matter at T13-L1 in the cat. Estimated conduction velocities in the spinally projecting axons ranged from 5.0-61 m/s. Stimulation of cell bodies in PGL, by microinjection of D,L-homocysteic acid, usually produced a rise in blood pressure accompanied by tachycardia or bradycardia and either dilatation or constriction in the hindlimb muscle vasculature. It is suggested from this other and evidence that tonically active neurones in PGL may play a role in setting the resting level of blood pressure but that the output of these and other cells in the nucleus may also be modified to generate different patterns of cardiovascular adjustment. PMID- 2989771 TI - [Two-channel bronchial artery infusion of cisplatin for the treatment of primary lung cancer]. PMID- 2989773 TI - [Differential diagnosis of small hepatic mass lesions by intra-arterial digital subtraction angiography of the time-density curves]. PMID- 2989772 TI - [Relative biological effectiveness of the therapeutic proton beams at NIRS and Tsukuba University]. PMID- 2989774 TI - Presence of a limited number of essential nucleotides in the promoter region of mouse ribosomal RNA gene. AB - Point mutations are introduced into a mouse rDNA fragment containing the promoter region by a sodium bisulfite method and the mutants are tested for the ability of accurate transcription initiation in vitro. The results indicate that the change, G to A, at -7 completely eliminates the promoter activity, and those at -16 and at -25 decrease it to about 10% and 50%, respectively. On the other hand, the substitutions at +9, +4, -2, -9 and -39 do not alter the template activity significantly. It is concluded that there are limited but distinct nucleotides that are essential for the transcription initiation of this gene. This sort of absolute requirement for single specific bases is not reported in protein coding genes transcribed by RNA polymerase II. We propose that these rigid recognition signals which we have found are the molecular basis for the strong species dependency of the transcription machinery of RNA polymerase I system. A model is presented in which a transcription factor interacts with the rDNA promoter from one side of the DNA double-helix with essential contacts at these bases. PMID- 2989775 TI - rRNA processing: removal of only nineteen bases at the gap between 28S alpha and 28S beta rRNAs in Sciara coprophila. AB - We have determined the sequence of the rDNA region between the 28S alpha and 28S beta rRNA coding segments (termed a "gap") in the insect Sciara coprophila, and have used S1 nuclease mapping and cDNA primer extension to define the 5' and 3' boundaries of the gap. Only 19 bases found in rDNA at the gap region are absent from mature 28S rRNA. Eukaryotic rRNAs contain stretches of nucleotides ("expansion segments") which are absent in E. coli rRNA. The gap region in Sciara is located within expansion segment V. Therefore, the excision of 19 bases in the Sciara gap suggests that a large portion of expansion segment V plays no function in mature ribosomes. Specific sequences conserved in Sciara and Drosophila are considered as candidates for recognition signals for the excision of the gap transcript. PMID- 2989776 TI - Overproduction of the EcoR V endonuclease and methylase. AB - Strains overproducing the EcoR V endonuclease and methylase have been obtained by inserting each of the two genes in expression vectors containing the lambda PL promoter. The methylase is overproduced to a level reaching 5-10% of the total cellular proteins, which represents a 50-100 fold increase. A 30 fold overproduction of endonuclease was achieved by randomly positioning the EndRV gene downstream of the lambda PL promoter. The situation in the endonuclease overproducing clone resembles that encountered in maxi-cells. The strains described here allowed a quick purification of both enzymes in sufficient amounts for crystallisation attempts. PMID- 2989777 TI - The primary structure of human hemopexin deduced from cDNA sequence: evidence for internal, repeating homology. AB - We have cloned and analyzed a cDNA containing the coding sequence for human hemopexin. We have first identified, by immunological screening of 30.000 colonies of a liver cDNA library in the expression vector pEX1, a clone carrying an insert 1170 base pairs long that shows 100% homology with a known human hemopexin peptide. The complete sequence coding for hemopexin was isolated from a liver cDNA library in the vector pAT218. The DNA insert of 1523 base pairs shows an open reading frame coding for 439 amino acids, a 3' noncoding region of 159 nucleotides long, followed by a poly(A) tail. The insert spans the entire coding region and from which the primary structure of the protein was deduced. By computer assisted analysis of the amino acid sequence, it was possible to recognize a core unit, of about 45 amino acids, which is repeated 8 or possibly even 10 fold along the polypeptide chain. This feature suggests that the gene might have evolved through a series of duplications. This characteristic, together with prediction of secondary structure, suggest a rough model for the tridimensional folding that allows some speculations on the function of hemopexin. Blot hybridization of total RNA from human liver with nick translated hemopexin cDNA detected a message of about 1600 nucleotides. Southern blot experiments to identify the hemopexin gene (s) suggest that it is not a large multi-gene family, but that there is only one or at most a few genes in the human genome. PMID- 2989778 TI - A new copia-like transposable element found in a Drosophila rDNA gene unit. AB - We have discovered a member of a new family of copia-like transposable elements inserted into the non-transcribed spacer between two ribosomal genes (rDNA). This family, which we call 3S18, consists of at least 15 elements which are scattered throughout the Drosophila melanogaster genome. The elements of this family are approximately 6.5 kb long and have 0.5 kb terminal direct repeats. All of the elements appear to have the same restriction sites. The element is mobile as the size pattern of homologous fragments varies among different strains. In situ hybridization results confirm the scattered location and transposable qualities of 3S18. The element is not transcribed into abundant RNA. PMID- 2989779 TI - Nucleotide sequence of the alpha ribosomal protein operon of Escherichia coli. AB - In Escherichia coli some 19 transcription units encoding the 52 ribosomal proteins are scattered throughout the genome. One of the units, the alpha operon, encodes genes for the ribosomal proteins S13, S11, S4 and L17 as well as the alpha subunit of RNA polymerase. We report here the complete 3.0 kb nucleotide sequence of the alpha operon. In addition, we have determined by S1 nuclease mapping the site of transcription termination in this operon. PMID- 2989780 TI - Pattern of undermethylation of the major satellite DNA of mouse sperm. AB - Enzymatic hydrolysis and base analysis by high performance liquid chromatography showed that mouse satellite DNA had 30-50% less 5-methylcytosine in sperm than in somatic tissue (1.59 mols % vs 2.40-3.11 mols %). Maxam-Gilbert sequencing and analysis of the intensity of the cytosine bands indicated that the level of methylation of the eight CpGs of the consensus sequence in sperm satellite DNA ranged from 0 to about 50%, considerably lower than the levels reported in somatic tissues. The Mn1I site containing one of these CpGs was cut much more extensively in satellite DNA from sperm than from liver, confirming the undermethylation of this site in sperm DNA. PMID- 2989781 TI - The ilvB locus of Escherichia coli K-12 is an operon encoding both subunits of acetohydroxyacid synthase I. AB - The ilvB locus of Escherichia coli K-12 encloses two open reading frames defining polypeptides of 60,000 and 11,200 molecular weight. The entire locus, about 2.3 kb, is co-transcribed as an operon. The molecular weights and amino acid compositions of the presumptive operon polypeptides agree with those of the large and small subunit polypeptides of acetohydroxyacid synthase (AHAS) I, for which ilvB is the structural locus. We reserve the designation ilvB for the promoter proximal (longer) cistron and designate the promoter distal cistron ilvN. The molecular weight and amino acid sequence of the ilvB polypeptide are strikingly similar to those of the I1vI (larger subunit of AHAS III) and I1vG (larger subunit of AHAS II) polypeptides. There is less size uniformity among the I1vN, I1vH (smaller subunit of AHAS III), and I1vM (smaller subunit of AHAS II) polypeptides. Nevertheless, there is significant amino acid sequence homology among the three small subunit polypeptides. Thus, all three AHAS isozymes of E. coli K-12 probably have a common evolutionary origin. PMID- 2989782 TI - The nucleotide sequence of the ilvBN operon of Escherichia coli: sequence homologies of the acetohydroxy acid synthase isozymes. AB - Three acetohydroxy acid synthase isozymes, AHAS I (ilvBN), AHAS II (ilvGM) and AHAS III (ilvIH) catalyze the first step of the parallel isoleucine-valine biosynthetic pathway in Escherichia coli. Previous DNA sequence and protein purification data have shown that AHAS II and AHAS III are composed of large and small subunits encoded in the ilvGMEDA and ilvIH operons, respectively. Recent protein purification and characterization data have demonstrated that the AHAS I isozyme is also composed of large and small subunits (L. Eoyang, L. and P. M. Silverman [1984] J. Bacteriol. 157:184-189). Now the complete DNA sequence of the operon encoding the AHAS I isozyme has been determined. These data show that both AHAS I subunits (Mr 60,400 and Mr 11,100) are encoded in this operon. The coordinant regulation of both genes of the ilvBN operon has also been demonstrated. Comparisons of the DNA sequences of the genes encoding all three AHAS isozymes have been performed. Conserved homologies were observed between both the large and small subunits of all three isozymes. The closest homology was seen between the AHAS I and AHAS II isozymes. On the basis of these comparisons a rationale for the evolution of the AHAS isozymes in E. coli has been proposed. PMID- 2989783 TI - Nucleotide sequence of the yeast ILV2 gene which encodes acetolactate synthase. AB - We have determined the nucleotide sequence of the yeast ILV2 gene which codes for the amino acid biosynthetic enzyme acetolactate synthase (ALS). ALS has recently been shown to be the target in bacteria, yeast and plants, of the potent new herbicide sulfometuron methyl. The coding sequence for the ILV2 polypeptide contains 2061 base pairs. Comparison of deduced amino acid sequences indicates considerable conservation between the yeast protein and the large subunits of the E. coli ALS II and ALS III isozymes. A major distinction between the three proteins is the presence of an additional 90 amino acids at the amino terminal of the yeast protein. The amino acid sequence in this region shows similarities to yeast mitochondrial transit sequences and may function as such, since yeast ALS is localized in the mitochondria. Consensus sequences for initiation and termination of transcription that are consistent with the ends of the ILV2 mRNA, as well as general amino acid control regulatory sequences have been identified. PMID- 2989784 TI - The ribosomal RNA genes of Drosophila mitochondrial DNA. AB - The nucleotide sequence of a segment of the mtDNA molecule of Drosophila yakuba which contains the A+T-rich region and the small and large rRNA genes separated by the tRNAval gene has been determined. The 5' end of the small rRNA gene was located by S1 protection analysis. In contrast to mammalian mtDNA, a tRNA gene was not found at the 5' end of the D. yakuba small rRNA gene. The small and large rRNA genes are 20.7% and 16.7% G+C and contain only 789 and 1326 nucleotides. The 5' regions of the small rRNA gene (371 nucleotides) and of the large rRNA gene (643 nucleotides) are extremely low in G+C (14.6% and 9.5%, respectively) and convincing sequence homologies between these regions and the corresponding regions of mouse mt-rRNA genes were found only for a few short segments. Nevertheless, the entire lengths of both of the D. yakuba mt-rRNA genes can be folded into secondary structures which are remarkably similar to secondary structures proposed for the rRNAs of mouse mtDNA. The replication origin containing, A+T-rich region (1077 nucleotides; 92.8% A+T), which lies between the tRNAile gene and the small rRNA gene, lacks open reading frames greater than 123 nucleotides. PMID- 2989785 TI - Multiple sequence-specific DNA binding activities are eluted from chicken nuclei at low ionic strengths. AB - DNA sequence-specific binding proteins eluted from chicken erythrocyte and thymus nuclei, and fractionated as described by Emerson and Felsenfeld (19), have been investigated by filter binding and footprint analyses. The erythrocyte nuclear protein fraction specifically binds to at least two sites within the 5' flanking chromatin hypersensitive site of the chicken beta A-globin gene, and to a site 5' to the human beta-globin gene. The major chicken beta A globin gene binding site [G)18CGGGTGG) and the human beta-globin gene binding site [TA)6(T)8C(T)4) occur at or near sequences which are hypersensitive to S1 nuclease cleavage in supercoiled plasmids. Downstream, the second chicken beta A-globin gene binding site includes the beta-globin gene CACCC consensus sequence. Filter binding studies also show other sequence specific binding activities to human N-ras and human (but not chicken) c-myc gene sequences. PMID- 2989787 TI - Variants within the yeast Ty sequence family encode a class of structurally conserved proteins. AB - The Ty transposable elements of Saccharomyces cerevisiae form a heterogeneous family within which two broad structural classes (I and II) exist. The two classes differ by two large substitutions and many restriction sites. We show that, like class I elements a class II element, Tyl-17, also appears to contain at least two major protein coding regions, designated TYA and TYB, and the organisational relationship of these regions has been conserved. The TYA genes of both classes encode proteins, designated p1 proteins, with an approximate molecular weight of 50 Kd and, despite considerable variation between the TYA regions at the DNA level, the structures of these proteins are remarkably similar. These observations strongly suggest that the p1 proteins of Ty elements are functionally significant and that they have been subject to selection. PMID- 2989786 TI - Cis and trans activation of adenovirus IVa2 gene transcription. AB - The transcriptional control region of the adenovirus IVa2 promoter was analyzed by cloning this promoter in front of a gene coding for bacterial chloramphenicol acetyl transferase (CATase) and estimating levels of CATase and IVa2 promoter specific RNA synthesized after transfection. To produce detectable amounts of CATase with the IVa2 promoter, an enhancer has to be present in cis. In the absence of enhancer sequences, the adenovirus E1A gene can not stimulate CATase synthesis. When cells were transfected with plasmids containing enhancer sequences and various IVa2 mutant promoters upstream of the CAT gene, we observed that CATase activity was not reduced significantly even after deletion of all sequences upstream of the RNA initiation site. Synthesis of IVa2 specific RNA was dependent on plasmids containing an enhancer (SV40 72 bp repeat) that was present in cis. In the absence of enhancer sequences, co-transfection to provide the adenovirus E1A gene in trans also stimulated IVa2 RNA synthesis. When HeLa cells were transfected with various deletion mutants with an enhancer in cis it was seen that sequences -38 to -64 base pairs upstream of the RNA initiation site are necessary for efficient transcription. The E1A gene in trans and an enhancer in cis have an additive effect on RNA synthesis from both IVa2 and major late promoters. The basis for the conflicting results between transcription and CATase synthesis is discussed. PMID- 2989788 TI - Codon-defined ribosomal pausing in Escherichia coli detected by using the pyrE attenuator to probe the coupling between transcription and translation. AB - This communication describes an assay for the relative translation efficiency of individual codons which makes use of the pyrE attenuator to probe the coupling between transcription and translation at the end of an artificial leader peptide. By cloning of short synthetic DNA fragments the codons to be tested were placed in the middle of the leader peptide and the downstream transcription of a pyrE"lacZ gene was monitored by measuring beta-galactosidase activity. The substitution, one by one, of three AGG codons for arginine with three CGT codons for the same amino acid residue was found to cause a two fold increase per codon of transcription over the pyrE attenuator, such that an eight fold higher frequency of pyrE expression was seen when all three AGG codons were replaced by CGT codons. No such effect of codon composition was observed, when the cells were grown with a low UTP pool which causes a reduction of the mRNA chain growth rate. PMID- 2989789 TI - Deletion of C-terminal amino acid codons of PhiX174 gene E: effect on its lysis inducing properties. AB - The lysis gene E of bacteriophage PhiX174 has been subjected to deletion and fusion analysis. Deletions of 11 to 90% of gene E specific nucleotides coding for to C-terminal region of the gene product were cloned under transcriptional control of lambda pL. For this purpose plasmid pSU1 was constructed which carries an extended polylinker region downstream of pL. Depending on the number of nucleotides after the last gene E specific codon, various C-terminal segments of protein E were replaced by 4, 5, 53 or 314 unrelated amino acids. Functional analysis for lysis inducing properties of the various gene E mutants revealed that the final 9 codons of the gene could be deleted without loss of function. However, replacements of 19 or more C-terminal codons eliminated gene E activity. Although the functional site of the gene E product is located within the N terminal half of the polypeptide, the C-terminal part of the protein appears to exhibit severe influence on conformation and/or regulation of the functional site. PMID- 2989791 TI - Physical and functional structure of a yeast plasmid, pSB3, isolated from Zygosaccharomyces bisporus. AB - The plasmid pSB3 of yeast Zygosacharomyces bisporus has been sequenced. It contains 6,615 base pairs, including a pair of inverted repeats (IR) consisting of 391 base pairs and 3 large open reading frames (ORF). One of the ORFs (A gene) participates in the recombination at the IRs and the other two (B and C genes) are necessary for the stable maintenance of this plasmid. The ARS sequence, which functions in a Saccharomyces cerevisiae host, was localized within 168 base pairs consisting of part of one of the IRs and a unique sequence contiguous to it. pSB3 can be maintained as stably in Z. rouxii as in the natural host Z.bisporus. In contrast, pSB3 is maintained fairly unstably in S.cerevisiae. The reason for this instability was found to be inefficient partitioning of pSB3 in S.cerevisiae. The molecular construction of pSB3 resembles that of 2-micron DNA, however, sequence homology at the DNA level was very poor. PMID- 2989790 TI - A bidirectionally active signal for termination of transcription is located between tetA and orfL on transposon Tn10. AB - A terminator of transcription with bidirectional activity has been located between the translation termination codons of the genes tetA and orfL on Tn10. These genes are transcribed towards each other. Each orientation of the intervening sequence is shown to reduce the expression of the lacZ and galK genes when cloned between the respective structural gene and its promotor. The 3'ends of the respective mRNAs were determined by S1 mapping. The results confirm that the same sequence capable of forming a stem-loop structure with a GC rich stem is the termination signal for both orientations. In the more efficient tetA orientation (99%-96% reduction of expression) this sequence is followed by a run of six thymines. In the less efficient orfL orientation (96%-78% reduction of expression) it is followed by an AT rich sequence with seven thymines out of eleven base pairs. PMID- 2989792 TI - Structure of a Neurospora RNA polymerase I promoter defined by transcription in vitro with homologous extracts. AB - A Neurospora in vitro transcription system has been developed which specifically and efficiently initiates transcription of a cloned Neurospora crassa ribosomal RNA gene by RNA polymerase I. The initiation site of transcription (both in vitro and in vivo) appears to be located about 850 bp from the 5' end of mature 17S rRNA. However, the primary rRNA transcripts are normally cleaved very rapidly at a site 120-125 nt from the 5' end in vitro and in vivo. The nucleotide sequence surrounding the initiation site has been determined. The region from -16 to +9 exhibits partial homology to the corresponding sequences from a wide variety of organisms including yeast, but the most striking similarity is to the initiation region from Dictyostelium discoideum which displays 73% homology to the Neurospora sequence from -23 to +47. The Neurospora sequences from -96 to +97 have been shown to be sufficient for transcription. This region contains two sequences displaying 8/9 bp matches to elements of the 5S rDNA promoter. PMID- 2989793 TI - Molecular cloning and the nucleotide sequence of cDNA to mRNA for non-neuronal enolase (alpha alpha enolase) of rat brain and liver. AB - The nucleotide sequence for alpha alpha enolase (non-neuronal enolase: NNE) of rat brain and liver was determined from recombinant cDNA clones. The sequence was composed of 1722 bp which included the 1299 bp of the complete coding region, the 108 bp of the 5'-noncoding region and the 312 bp of the 3'-noncoding region containing a polyadenylation signal. In addition, the poly(A) tail was also found. A potential ribosome-binding site was located 30 nucleotides upstream to the initiation codon in the 5'-noncoding region. The amino acid sequence deduced from the nucleotide sequence was 433 amino acids in length and showed very high homology (82%) to the amino acid sequence of gamma gamma enolase (neuron-specific enolase: NSE), although the nucleotide sequence showed slightly lower homology (75%). The size of NNE mRNA was approximately 1800 bases by Northern transfer analysis and much shorter than that of NSE mRNA (2400 bases) indicating a short 3'-noncoding region. A dot-blot hybridization and Northern transfer analysis of cytoplasmic RNA from the developing rat brains using a labeled 3'-noncoding region of cDNA (no homology between NSE and NNE) showed a decrease of NNE mRNA at around 10 postnatal days and then a gradual increase to adult age without changes of mRNA size. Liver mRNA did not show any significant change during development. PMID- 2989794 TI - Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression. AB - Tumour necrosis factor (TNF), released by induced macrophages, causes tumour necrosis in animals and kills preferentially transformed cells in vitro. mRNA induced in the established mouse monocytic PU 5.1.8 cell line by lipopolysaccharide, was converted into double-stranded cDNA and cloned in the pAT153 vector. Recombinant plasmids were screened by plus-minus hybridization and TNF-specific oligonucleotide probes constructed on the basis of partial amino acid sequences of rabbit TNF. A series of TNF specific clones were identified and confirmed by hybrid selection of mouse TNF-specific mRNA. The sequence codes for a 235 amino acids long polypeptide, of which 156 amino acids presumably correspond to the mature product. It can be concluded that mature mouse TNF is a glycosylated dimer. Biologically active TNF was secreted by both Cos-I and CHO cells transfected with the chimaeric expression vector pSV2d2-mTNF containing the coding region of the mouse TNF cDNA gene. PMID- 2989795 TI - Improved oligonucleotide site-directed mutagenesis using M13 vectors. AB - An improved method is described for the construction of mutations in M13 vectors using synthetic oligonucleotides. The DNA is first cloned into a novel M13 vector (based upon M13mp18 or M13mp19), which carries a genetic marker that can be selected against, such as an EcoK or EcoB site, or an amber mutation in an essential phage gene. In this "coupled priming" technique, one primer is used to construct the silent mutation of interest, and a second primer is used to eliminate the selectable marker on the minus strand. After primer extension and ligation, the heteroduplex DNA is transfected into a strain of E. coli which is repair deficient and selects against the plus strand marker. Over 50 mutants have been constructed with this approach, and the yields can be excellent (up to 70%). For the stepwise construction of mutations using separate rounds of mutagenesis, the EcoK and EcoB markers offer a particular advantage over the amber marker. They permit selection in each round, as it is possible to cycle between the two markers. However for construction of multiple mutations over a short region, long synthetic oligonucleotides with multiple mismatches to the template can offer an alternative strategy. PMID- 2989796 TI - Structural junctions in DNA: the influence of flanking sequence on nuclease digestion specificities. AB - When a protein binds to DNA, the affinity of this protein for its primary site of interaction may be influenced by the nature of flanking sequences. This is thought to be a consequence of local cooperativity in the DNA molecule, where the conformation at one point along the helix can influence the conformation at another, and thereby modulate the free energy of protein-DNA recognition. In order to learn more about this process, we have carried out experiments of two sorts. First, we have constructed sequences of the type (dA)11 (dG)8, where the conformational preferences of the DNA molecule switch from one extreme to another over just a single base pair, and subjected them to digestion by DNAase I and DNAase II. This is to learn whether the structure changes abruptly at the junction point, or more gradually with an influence extending into residues on either side. Secondly, we have subjected long plasmid DNA to digestion by restriction enzymes Fnu DII, Hae III, Hha I and Msp I, to look for correlations between cutting rate and the identity of nucleotides on either side of the restriction site. The influence of flanking sequence on nuclease digestion specificities is clearly evident in both kinds of experiment, but the rules governing this seem complex and not easily formulated. The best that can be done at present is to divide the problem into two parts, "analogue" and "digital", representing sugar-phosphate and base components of recognition. PMID- 2989797 TI - Human P1-450 gene sequence and correlation of mRNA with genetic differences in benzo[a]pyrene metabolism. AB - The human P1-450 gene (6,311 base pairs), as well as the 5' (1,604 bases) and 3' (113 bases) flanking regions, have been completely sequenced. Four highly homologous boxes (61, 82, 56 and 97 base pairs) between the human and mouse P1 450 genes are found in the "TATA" box promoter region, -226, -338, and -450 upstream from the cap site, respectively. Nine genomic-DNA samples were digested with each of 23 restriction endonucleases and probed with human P1-450 cDNA fragments; restriction fragment length polymorphisms are detected, although it remains to be seen whether such a recombinant DNA test will be useful in determining individuals at increased risk for cigarette smoking-induced cancer and toxicity. We show in this report, however, that human inducible P1-450 mRNA concentrations are very highly correlated (r = 0.98; N = 6) with genetic differences in benzo[a]pyrene metabolism in mitogen-activated lymphocyte cultures. PMID- 2989798 TI - Sequence of a pseudogene in the legumin gene family of pea (Pisum sativum L.). AB - A second legumin gene, denoted psi Leg D, has been located on the pea genomic clone lambda Leg 1, approx. 1.3 Kbases 3' of Leg A, in the same orientation. The complete sequence of psi Leg D shows that it is a pseudogene, having two stop codons near the 5' end of its predicted coding sequence, as well as deletions and frame shift errors when compared to Leg A. No transcripts from this gene could be detected in developing pea seeds. Leg A and psi Leg D are homologous over their coding sequences, and partially homologous in the intron sequences and the immediate 5' flanking sequences. Other flanking sequences of the two genes show no significant homology, apart from the presence of polyadenylation signals 3' to both coding sequences. The introns in the two genes occur in corresponding positions in the sequences, but a deletion in psi Leg D affects the 3' boundary of IVS-2. Hybridisation of psi Leg D to pea genomic DNA suggests that it does not represent a hitherto undetected sub-family of legumin genes. PMID- 2989800 TI - The human apolipoprotein A-II gene: complete nucleic acid sequence and genomic organization. AB - The gene for human apolipoprotein (apo) A-II has been isolated from a human genomic DNA library. The cloned fragment was approximately 14 kilobase-pair (kb) long, and extended about 9.0 kb upstream as well as 3.5 kb downstream from the apoA-II gene, which was contained within a 3.1 kb HindIII fragment of human DNA. The complete nucleic acid sequence of the apoA-II gene has been determined, establishing that the apoA-II gene is interrupted by three intervening sequences of 182, 293, and 395 bp. The second intron is of particular interest, because it contains a 33 bp sequence of alternating G and T residues very close to the 3' splice site which has the potential to form a left handed Z-helix structure in vivo. A restriction fragment length polymorphism 3' from the apoA-II gene has been detected which may serve as a marker for the long arm of chromosome 1 in linkage analyses. PMID- 2989801 TI - Mental health. Practice makes perfect. PMID- 2989799 TI - Nitrogen fixation specific regulatory genes of Klebsiella pneumoniae and Rhizobium meliloti share homology with the general nitrogen regulatory gene ntrC of K. pneumoniae. AB - We have determined the complete nucleotide sequences of three functionally related nitrogen assimilation regulatory genes from Klebsiella pneumoniae and Rhizobium meliloti. These genes are: 1) The K. pneumoniae general nitrogen assimilation regulatory gene ntrC (formerly called glnG), 2) the K. pneumoniae nif-specific regulatory gene nifA, and 3) an R. meliloti nif-specific regulatory gene that appears to be functionally analogous to the K. pneumoniae nifA gene. In addition to the DNA sequence data, gel-purified K. pneumoniae nifA protein was used to determine the amino acid composition of the nifA protein. The K. pneumoniae ntrC and nifA genes code for proteins of 52,259 and 53,319 d respectively. The R. meliloti nifA gene codes for a 59,968 d protein. A central region within each polypeptide, consisting of approximately 200 amino acids, is between 52% and 58% conserved among the three proteins. Neither the amino termini nor the carboxy termini show any conserved sequences. Together with data that shows that the three regulatory proteins activate promoters that share a common consensus sequence in the -10 (5'-TTGCA-3') and -23 (5'-CTGG-3') regions, the sequence data presented here suggest a common evolutionary origin for the three regulatory genes. PMID- 2989802 TI - [Comparison of the results of surgical and conservative treatment of small-cell lung cancer]. PMID- 2989803 TI - [Comparison of the effects of Kenalog and Zaditen on the clinical state and plasma cAMP levels of patients with bronchial asthma]. PMID- 2989804 TI - Non-streptococcal pharyngitis. PMID- 2989806 TI - [Multifocal cancer of the large intestine]. PMID- 2989805 TI - [Effect of stimulation of beta-adrenergic and H2 receptors of polymorphonuclear granulocytes on the release of lysosomal enzymes in patients with hay fever before and after treatment with Pollinex]. PMID- 2989807 TI - [Calmodulin]. PMID- 2989808 TI - Bowenoid papulosis of the genitalia associated with human papillomavirus DNA type 16 in an infant with atopic dermatitis. AB - A case of bowenoid papulosis occurred in a 2-year-old male with atopic dermatitis. Clinical and histologic features of the lesions were typical, and human papillomavirus type 16 DNA was identified using high-stringency hybridization techniques. Although the lesions had been present for approximately one year prior to examination, they subsequently resolved spontaneously over six months. We postulate that our patient's susceptibility to human papillomavirus may have been related to his severe atopic tendency. PMID- 2989809 TI - Diet, nutrition, and cancer. An update on a controversial relationship. AB - The multiple-step model of carcinogenesis discussed here identifies the two major stages of initiation and promotion. A more recent research development proposes that oncogenes present in chromosomes are activated by viral, chemical, or physical agents and cause cancer. A great variety of natural mutagens and carcinogens find their way into the modern US diet. Excessive fat and alcohol consumption have been studied in relation to many kinds of malignancies. Dietary anticarcinogens include vitamins A, C, and E, although under certain conditions some generally inhibitive substances can actually enhance carcinogenesis. A provocative hypothesis argues that a high-fiber diet can substantially reduce the likelihood of carcinoma of the colon. PMID- 2989810 TI - Chronic heat stress and respiratory alkalosis: occurrence and treatment in broiler chicks. AB - The occurrence of respiratory alkalosis and potential benefit derived from treatment were examined in thermostressed 4-week-old broiler chicks. Blood pH was greater (P less than .05) in heat-stressed (32 C) panting birds (7.395) than either nonpanting (7.28) or birds raised at 24 C (7.28). Acute thermostress, obtained by elevating ambient temperature from 32 to 41 C over a 20-min period further elevated (P less than .05) blood pH to 7.521. Chronic heat-stressed broiler chicks suffer from intermittent respiratory alkalosis during panting; with acute heat stress, chicks pant continuously and suffer from alkalosis. Including .5% sodium bicarbonate (NaHCO3) in the diet of birds subjected to chronic heat stress enhanced body weight gain by 9% even though it tended (P less than .10) to increase blood pH in nonpanting birds. Adding .3 or 1% ammonium chloride (NH4Cl) to diets decreased blood pH (P less than .01) to 7.194 and increased (P less than .05) body weight gains by 9.5 and 25%, respectively. Effects appeared linear with NH4Cl dose to 1% NH4Cl, but 3% NH4Cl elevated weight gains by only 8% and precipitated blood acidosis (pH 7.09) in nonpanting birds. Supplementing the 1% NH4Cl diet with .5% NaHCO3 increased weight gains an additional 9%. Manipulating sodium: chloride ratios by addition of calcium chloride increased body weight gain 8% and slightly reduced severity of alkalosis. Data indicate that blood alkalosis limits growth rate of broiler chicks reared under chronic thermostress and that the respiratory alkalosis and weight gain depressions attributed to thermostress can be partially alleviated dietarily. PMID- 2989811 TI - Shell quality as influenced by zeolite with high ion-exchange capability. AB - Sixteen dietary treatments applied to a total of 960 hens were used to determine the influence of zeolite A on shell quality and egg size. In Experiment 1, sodium zeolite A (SZA) was fed at three levels (0, .75, and 1.50%) in diets containing 4.0 and 2.75% calcium (Ca) for 8 weeks to old hens. In Experiment 2, the same levels of SZA were fed in diets containing two total sulfur amino acid levels (TSAA, .61 and .51%) to young hens for 12 weeks. Calcium zeolite A (CZA) was also fed at .68% in Experiment 1 in the 4.0% Ca diets and in Experiment 2 in the .51 and .61% TSAA diets. These diets were adjusted for sodium (Na) and chloride (Cl). SZA (.75% unadjusted for Na and Cl) was fed to old hens receiving the 2.75% Ca diet in Experiment 1. All diets were isocaloric and isonitrogenous within diets having the same Ca or TSAA level within an experiment. Response criteria were egg production, feed consumption, egg specific gravity, serum Ca, and body weight. A significant linear response in egg specific gravity occurred within 2 or 3 weeks, when diets supplemented with SZA were fed to old (Experiment 1) and young (Experiment 2) hens. Average Ca intake for control hens (Experiment 1) fed the 2.75 and 4.0% Ca diets was 2.93 and 4.54 g, respectively. Average Ca intake for control hens (Experiment 2) fed the .51 and .61% TSAA diet was 4.38 and 4.00 g, respectively. The CAZ (Experiments 1 and 2) and SZA (unadjusted for Na and Cl, Experiment 1) also gave significant increases in egg specific gravity. Zeolite A had little or no influence on egg weight, feed consumption, or egg production in Experiments 1 or 2. When Na and Cl were not adjusted in the SZA treatments (Experiment 1) a significant reduction in production occurred. It was concluded that zeolite A will significantly increase egg specific gravity and we hypothesize that the mechanism responsible for the significant improvement is related to the high ion-exchange capability of zeolite A. PMID- 2989812 TI - Intracranial space-occupying lesions in patients attending a migraine clinic. PMID- 2989813 TI - GTPase of bovine rod outer segments: the amino acid sequence of the alpha subunit as derived from the cDNA sequence. AB - The sequence of the 350 amino acids in the alpha subunit of GTPase of bovine rod outer segments has been determined. Enriched GTPase mRNA was used to prepare a cDNA library in the expression vector lambda gt11 and several overlapping cDNA clones corresponding to the alpha subunit of the GTPase were identified. The cDNA sequence determined contains 93 nucleotides upstream of the 5' end of the coding region, 1050 nucleotides that specify the amino acid sequence, and 45 nucleotides downstream from the 3' end. The previously described partial amino acid sequences and the sequences at the ADP-ribosylation sites for cholera and pertussis toxins are all confirmed and fitted into the present complete sequence. Homologies are found between the sequence of the alpha subunit and those of other guanine nucleotide-binding proteins, the ras proteins, peptide chain elongation factors EF-Tu and EF-G, and the initiation factor IF2. PMID- 2989814 TI - Adenovirus VAI RNA prevents phosphorylation of the eukaryotic initiation factor 2 alpha subunit subsequent to infection. AB - The virus-associated VAI RNA of adenovirus is a small, RNA polymerase III transcribed species required for efficient translation of mRNAs late after infection. Deletion mutant dl331 fails to produce this RNA and, as a result, grows poorly. Three lines of evidence suggest that VAI RNA facilitates translation by preventing inactivation of the function of eukaryotic initiation factor 2 (eIF-2). First, the mutant's translational defect can be relieved by addition of eIF-2 or eIF-2B (GTP recycling factor). Second, extracts of mutant infected cells exhibit enhanced protein P1/eIF-2 alpha subunit kinase activity. Third, dl331 can grow with nearly normal kinetics in cells that do not express the kinase. PMID- 2989815 TI - Expression of a functional influenza viral cap-recognizing protein by using a bovine papilloma virus vector. AB - The gene for the influenza viral PB2 protein, which recognizes and binds the 5' terminal cap 1 structures (m7GpppNm) on eukaryotic mRNAs, was inserted into a bovine papilloma virus vector under the control of a mouse metallothionein I (MT I) promoter. After transfection of this vector into mouse NIH 3T3 cells, a cell line, cPB2, was obtained that produces PB2-specific mRNA and authentic PB2 protein. Induction of the MT-I promoter with CdCl2 causes about a 10-fold increase in PB2 mRNA and protein levels. The expressed PB2 protein is functional, as it relieves the block in viral mRNA synthesis exhibited by a temperature sensitive viral mutant containing a cap-binding defect in the PB2 protein. This demonstrates complementation of a function of a negative-strand RNA virus by a gene product expressed in a cell line from recombinant DNA. PMID- 2989816 TI - umuDC and mucAB operons whose products are required for UV light- and chemical induced mutagenesis: UmuD, MucA, and LexA proteins share homology. AB - The products of the Escherichia coli umuDC operon and its plasmid-borne analog, mucAB, are required for mutagenesis caused by UV light and by many chemicals. We have determined the nucleotide sequences of umuDC and mucAB and present comparisons of these sequences. The two operons are 52% homologous at the nucleotide level. Open reading frames corresponding in position and size to the umu and muc genes have been identified. The reading frames of umuD and umuC overlap by 1 base pair, and the reading frames of mucA and mucB overlap by 13 base pairs. The predicted amino acid sequences of the UmuD and MucA proteins are 41% homologous; those of the UmuC and MucB proteins are 55% homologous. Considerable homology has also been detected between UmuD, MucA, and the COOH terminal domains of the LexA repressor and the repressors of phage lambda, 434, and P22. Complementation analyses reveal that MucA protein cannot substitute for UmuD in a umuD- umuC+ host and that MucB protein cannot substitute for UmuC in a umuD+ umuC- host. Potential regulatory sequences have been identified in umuDC and mucAB. PMID- 2989817 TI - Structural analysis of the umu operon required for inducible mutagenesis in Escherichia coli. AB - The nucleotide sequence of a 2.8-kilobase fragment of the Escherichia coli chromosome containing the umuDC genes has been determined. The DNA sequence specifies two open reading frames of 417 and 1266 nucleotides encoding proteins with calculated molecular weights of 15,063 and 47,677, respectively. From these and the previous results of genetic and biochemical studies on the cloned genes, we conclude that the former is the umuD and the latter is the umuC gene. In vitro transcription of the regulatory region of the umu operon revealed that (i) upstream of the coding region there is a promoter-operator complex having the consensus sequence, CTGTATATAAAAACAG, of an SOS box, (ii) transcription of the umu operon begins at an adenine residue in the SOS box, and (iii) the LexA protein binds to the operator region, with an equilibrium dissociation constant (Kd) of 0.2, thereby repressing the transcription of the umuDC genes. These results indicate that the expression of the umu operon is under the coordinated control of the recA lexA gene products. PMID- 2989818 TI - Cloning, characterization, and sequence of the yeast DNA topoisomerase I gene. AB - The structural gene for yeast DNA topoisomerase I (TOP1) has been cloned from two yeast genomic plasmid banks. Integration of a plasmid carrying the gene into the chromosome and subsequent genetic mapping shows that TOP1 is identical to the gene previously called MAK1. Seven top1 (mak1) mutants including gene disruptions are viable, demonstrating that DNA topoisomerase I is not essential for viability in yeast. A 3787-base-pair DNA fragment including the gene has been sequenced. The protein predicted from the DNA sequence has 769 amino acids and a molecular weight of 90,020. PMID- 2989819 TI - Identification of an inhibitory region of the heat-stable protein inhibitor of the cAMP-dependent protein kinase. AB - The present study was undertaken in order to identify the inhibitory site of the heat-stable inhibitor of cAMP-dependent protein kinase (PKI) and to synthesize a peptide that could serve as a useful inhibitor of the enzyme. Digestion of purified PKI by mast cell proteinase II yielded a peptide fragment that retained inhibitory activity. A sequence of 20 amino acids of the peptide, (sequence in text) revealed the presence of a "pseudosubstrate site" (Arg-Arg-Asn-Ala-Ile) for the cAMP-dependent protein kinase in which alanine replaces the seryl or threonyl residue that is normally phosphorylated. Digestion of PKI with various other proteinases implicated the involvement of arginyl and hydrophobic residues as determinants for the inhibitory activity. The assumption that this region is part of the inhibitory site was confirmed by the synthesis of a corresponding duodecapeptide that displayed strong inhibitory activity. Inhibition by the peptide was competitive with a Ki of 0.8 microM as measured against a number of protein substrates. The sequence of this fragment bears a strong resemblance to the autophosphorylation site in the type II regulatory subunit of cAMP-dependent protein kinase, a region also postulated to interact with the catalytic subunit, and the analogous region of type I regulatory subunit. Neither intact PKI nor the synthetic peptide inhibit the cGMP-dependent protein kinase, phosphorylase kinase, myosin light-chain kinase, casein kinase II, or protein kinase C. PMID- 2989820 TI - Arterial smooth muscle cells in primary culture produce a platelet-derived growth factor-like protein. AB - Adult rat arterial smooth muscle cells (SMC) in primary culture modulate from contractile to synthetic phenotype. This process includes partial loss of myofilaments and formation of an extensive rough endoplasmic reticulum and a large Golgi complex. It gives the cells the ability to initiate DNA synthesis and actively proliferate when stimulated with serum or isolated growth factors. After a few divisions, growth becomes partly independent of exogenous mitogens and does not cease until multiple cell layers have been formed. Here, it is demonstrated that serum-free conditioned medium from primary cultures of adult rat arterial SMC contains a factor that initiates DNA synthesis in growth-arrested secondary cultures of SMC. The mitogenic activity was neutralized by antibodies to platelet derived growth factor (PDGF), and no mitogenic activity occurred in conditioned medium from cultures pretreated with actinomycin D, excluding release into the medium of PDGF adsorbed to the plastic vessels during the initial culture in serum-containing medium. Exposure of human fibroblasts to samples of the conditioned medium at 4 degrees C inhibited subsequent binding of 125I-labeled PDGF. It was further shown that the SMC of the primary cultures were able to initiate DNA synthesis in a chemically defined medium lacking PDGF and other growth factors. During the early, most active, and partly autonomous growth phase, the SMC had a low binding capacity for 125I-labeled PDGF and responded but little to stimulation with exogenous PDGF. Later on, with increasing cell density and decreasing growth rate, the ability to bind and respond to exogenous PDGF increased. Taken together, the observations suggest that modulation of SMC from contractile to synthetic phenotype is accompanied by production of a PDGF-like protein and autocrine or possibly by mitogen-independent initiation of DNA synthesis. Functionally, this may be important during wound healing and in the development of atherosclerotic lesions. PMID- 2989821 TI - Human urokinase gene is located on the long arm of chromosome 10. AB - Urokinase is one of the two plasminogen activators that catalyze the conversion of inactive plasminogen to plasmin. By combining somatic cell genetics, in situ hybridization, and Southern hybridization, we localized the human urokinase gene on the distal third of the long arm (q24-qter) of chromosome 10. PMID- 2989822 TI - Hepatitis B virus integration in hepatocellular carcinoma DNA: duplication of cellular flanking sequences at the integration site. AB - The integrated form of hepatitis B virus (HBV) in the human hepatoma cell line huH2-2 and its cellular counterpart sequence have been cloned and analyzed. Blot hybridization analysis and nucleotide sequencing indicated that a single copy of the 1895-base-pair (bp) subgenomic region of HBV DNA, spanning from the middle of pre-S to the end of gene X, was integrated and flanked by the 12-bp directly repeating cellular sequences. A comparison of the sequencing data with that of the cellular counterpart DNA indicated the absence of deletion and rearrangement in the cellular flanking DNA following integration of the 1895-bp HBV DNA, except for generation of the 12-bp direct repeat at the virus-cell junction. A possible model for the mechanism of HBV integration is proposed. PMID- 2989823 TI - Nucleotide sequence of the Dpn II DNA methylase gene of Streptococcus pneumoniae and its relationship to the dam gene of Escherichia coli. AB - The structural gene (dpnM) for the Dpn II DNA methylase of Streptococcus pneumoniae, which is part of the Dpn II restriction system and methylates adenine in the sequence 5'-G-A-T-C-3', was identified by subcloning fragments of a chromosomal segment from a Dpn II-producing strain in an S. pneumoniae host/vector cloning system and demonstrating function of the gene also in Bacillus subtilis. Determination of the nucleotide sequence of the gene and adjacent DNA indicates that it encodes a polypeptide of 32,903 daltons. A putative promoter for transcription of the gene lies within a hundred nucleotides of the polypeptide start codon. Comparison of the coding sequence to that of the dam gene of Escherichia coli, which encodes a similar methylase, revealed 30% of the amino acid residues in the two enzymes to be identical. This homology presumably reflects a common origin of the two genes prior to the divergence of Gram-positive and Gram-negative bacteria. It is suggested that the restriction function of the gene is primitive, and that the homologous restriction system in E. coli has evolved to play an accessory role in heteroduplex DNA base mismatch repair. PMID- 2989824 TI - Close linkage of alpha and beta interferons and infrequent duplication of beta interferon in humans. AB - Five restriction fragment length polymorphisms in the human alpha/beta interferon (IFN-alpha/beta) gene region were identified, three with an IFN-alpha probe and two with an IFN-beta probe. Heterozygosities are 74% for IFN-alpha, 57% for IFN beta, and 87% jointly, making IFN-alpha/beta genes excellent markers for the short arm of chromosome 9. The pedigrees of about 25 families of Caucasian background were studied. Segregation analysis disclosed the occurrence of 12 of 32 possible haplotypes. No recombinant was found between IFN-alpha and -beta genetic markers; linkage disequilibrium within the IFN-alpha markers is of a similar order of magnitude as that between the IFN-alpha and -beta markers. The IFN-alpha and -beta genes might cluster within several hundred kilobases. In two parents, the IFN-beta gene is duplicated; the duplications segregate regularly. They are of independent, probably recent, origin. Therefore, some degree of multiplicity might even be found for IFN-beta, at least in some individuals. PMID- 2989825 TI - Domain structure and associated functions of subcomponents C1r and C1s of the first component of human complement. AB - The serine protease subcomponents of the activated form of the first component of human complement (C1), C1r and C1s, were observed by electron microscopy after the native proteins and their limited proteolysis products, obtained from autolytic cleavage (C1r) or from incubation with plasmin (C1s) were rotary shadowed. At the monomeric level, both C1r and C1s comprised two globular domains, a smaller interaction domain (corresponding to the NH2-terminal half of the A chain, alpha, and responsible for calcium binding and C1r-C1s interaction) and a larger catalytic domain (corresponding to the COOH-terminal part of the A chain, gamma, disulfide-linked to the B chain and bearing the serine protease active site). The two globular domains are linked by a connecting strand, beta. The (C1r)2 dimer appeared as a "croissant"-like association, where the two monomers interact through their catalytic domains. On the basis of the domain structure of C1r and C1s, a model of the calcium-dependent C1s dimer is proposed, in which the two monomers interact through their NH2-terminal interaction domains; in the same way, a model of the C1s-(C1r)2-C1s catalytic subunit of C1 is presented, in which (C1r)2 forms a core, its distal interaction domains interacting with the corresponding domains of C1s. PMID- 2989826 TI - Identification of genes for the constant region of rabbit T-cell receptor beta chains. AB - We describe cDNA clones from thymus mRNA of a young rabbit that have sequences highly homologous to the human and murine T-cell receptor beta-chain constant region (C beta). In rabbit, man, and mouse there is a conserved extra cysteine in the constant region that could lead to a free thiol group or alternative disulfide bond formation depending on the locations and total numbers of cysteines in assembled receptor molecules. A cDNA clone (CL ANA 11) with 571 bases 5' of the C beta coding sequence has an open reading frame starting at a methionine codon that encodes 141 amino acids in frame with the C beta sequence. The encoded sequence has no resemblance to known immunoglobulin or beta-chain variable regions or other known proteins. An oligonucleotide probe from the 5' end of the encoded protein hybridizes to an approximately equal to 2-kilobase genomic DNA fragment that contains C beta gene sequences and to an approximately equal to 8-kilobase mRNA species in the thymus mRNA preparation from which the clone was derived. Within the 5' coding sequence there is a stretch of 211 bases containing strings of alternating purines and pyrimidines that may form Z-DNA. The sequence of the last 55 base pairs adjacent to C beta resembles the corresponding segment of mouse cDNA clone 86T3 that contains sequence 5' of the mouse C beta 1 gene. Although the function of a potential protein encoded by the 5' end of CL ANA 11 is unknown, it could play a role in regulation of thymocyte growth and differentiation. PMID- 2989827 TI - Generation and regulation of autocytotoxicity in mixed lymphocyte cultures: evidence for active suppression of autocytotoxic cells. AB - Using limiting dilution analysis, we have detected both the generation and suppression of autocytotoxic cells following autologous or allogeneic stimulation in vitro. Assay conditions were established in which the cytotoxic response toward an allogeneic sensitizing cell was consistent with a traditional single hit kinetic model. Under identical conditions, cytolytic activity toward autologous phytohemaglutinin-activated lymphoblasts exhibited a distinct biphasic response. At low responder cell doses, a clear autocytotoxic response was observed. However, at higher responder cell numbers, this autocytotoxic reaction disappeared. This biphasic pattern of autocytotoxicity developed after stimulation with allogeneic or autologous peripheral blood mononuclear leukocytes (PBL) or Epstein-Barr virus-transformed B cells. This pattern of response is consistent with the counterpoised actions of two distinct cell populations, an autoaggressive population and a lower frequency autosuppressor population. Autocytotoxicity was not the result of mitogenic or xenogeneic antigenic stimulation, as it was observed after stimulation with autologous PBL in autologous serum and an autologous interleukin 2 preparation. Thus, cells capable of autocytotoxicity are present in peripheral blood but at a lower frequency than allocytotoxic T lymphocytes. Furthermore, autoaggressive cells are down-regulated by an autologous suppressor population. These findings suggest that immunologic self-tolerance is, at least in part, an actively maintained condition. Disturbances in this autoregulatory network may have relevance to the pathogenesis of some autoimmune diseases and graft-versus-host disease. PMID- 2989828 TI - Deficiency of electron transfer flavoprotein or electron transfer flavoprotein:ubiquinone oxidoreductase in glutaric acidemia type II fibroblasts. AB - Glutaric acidemia type II (GA II) is a human genetic disorder. It has been suggested that the primary defect in this disorder is a deficiency of a protein involved in electron transport between the acyl-CoA dehydrogenases and the bc1 complex of the mitochondrial respiratory chain. Antisera were raised to purified porcine electron transfer flavoprotein (ETF) and electron transfer flavoprotein:ubiquinone oxidoreductase (ETF:QO). The antisera were used to detect the two electron transferases in control and GA II fibroblasts by immunoblotting. Fibroblasts from three unrelated GA II patients were deficient in immunologically detectable ETF:QO and extracts from these three fibroblast lines contained no detectable ETF:QO catalytic activity. Fibroblasts from parents of two of these patients had ETF:QO activity intermediate between activities in control fibroblasts and fibroblasts from the patients. These data indicate that the primary defect in these patients is a deficiency of ETF:QO and that the mode of transmission of the gene is autosomal recessive. Fibroblasts from two other patients with severe GA II had normal levels of ETF-QO activity and antigen but were deficient in immunoreactive ETF. These findings show that GA II results from a deficiency of ETF in some patients and ETF:QO in others. In addition, these investigations provide strong evidence for the specificity and physiological function of the iron-sulfur flavoprotein ETF:QO. PMID- 2989829 TI - Beta 2-adrenergic receptors contribute to catecholamine-stimulated shortening of action potential duration in dog atrial muscle. AB - beta 1- and beta 2-adrenergic receptors coexist in atria of humans and a variety of other species. The significance of these receptors and the quantitative extent to which beta 2-adrenergic receptors contribute to the electrophysiologic effects of catecholamines acting at beta-adrenergic receptors have not been determined. In the present study, the beta 1-selective antagonist ICI 89,406 and the beta 2 selective antagonist ICI 118,551 were used to determine the relative densities of beta 1- and beta 2-adrenergic receptors on membranes prepared from dog atria and the contribution that each subtype makes to isoproterenol-induced shortening of action potential duration measured at 75% repolarization (APD75). Computer-aided nonlinear regression analysis of the inhibition of the specific binding of [125I]iodopindolol and the inhibition of isoproterenol-stimulated APD75 shortening by these antagonists showed that a two-site model fits the data better than a one-site model (P less than 0.0001). The affinity constants for each selective antagonist determined by inhibition of APD75 shortening were similar to those determined in studies of the inhibition of the specific binding of [125I]iodopindolol to beta 1- and beta 2-adrenergic receptors. beta 2-Adrenergic receptors made up approximately equal to 25% of the total number of beta adrenergic receptors but mediated approximately equal to 50% of the electrophysiologic effect of isoproterenol. The inhibition of the binding of [125I]iodopindolol and of the APD75 shortening by propranolol, a nonselective antagonist, was best fit by a one-site model. In other experiments, dogs were treated with reserpine and atropine to eliminate complications caused by the presence of endogenous norepinephrine and acetylcholine. Results obtained with atria from these animals were similar to those obtained in studies with control atria. These data suggest that beta 2-adrenergic receptors as well as beta 1 adrenergic receptors are involved in mediating the electrophysiologic effects of catecholamines in dog atrial muscle and that both beta 1- and beta 2-adrenergic receptors are present on atrial muscle cells. PMID- 2989830 TI - Dietary saturated triacylglycerols suppress hepatic low density lipoprotein receptor activity in the hamster. AB - The liver plays a key role in the regulation of circulating levels of low density lipoproteins (LDL) because it is both the site for the production of and the major organ for the degradation of this class of lipoproteins. In this study, the effects of feeding polyunsaturated or saturated triacylglycerols on receptor dependent and receptor-independent hepatic LDL uptake were measured in vivo in the hamster. In control animals, receptor-dependent LDL transport manifested an apparent Km value of 85 mg/dl (plasma LDL-cholesterol concentration) and reached a maximum transport velocity of 131 micrograms of LDL-cholesterol/hr per g, whereas receptor-independent uptake increased as a linear function of plasma LDL levels. Thus, at normal plasma LDL-cholesterol concentrations, the hepatic clearance rate of LDL equaled 120 and 9 microliter/hr per g by receptor-dependent and receptor-independent mechanisms, respectively. As the plasma LDL-cholesterol was increased, the receptor-dependent (but not the receptor-independent) component declined. When cholesterol (0.12%) alone or in combination with polyunsaturated triacylglycerols was fed for 30 days, receptor-dependent clearance was reduced to 36-42 microliter/hr per g, whereas feeding of cholesterol plus saturated triacylglycerols essentially abolished receptor dependent LDL uptake (5 microliter/hr per g). When compared to the appropriate kinetic curves, these findings indicated that receptor-mediated LDL transport was suppressed approximately equal to 30% by cholesterol feeding alone and this was unaffected by the addition of polyunsaturated triacylglycerols to the diet. In contrast, receptor-dependent uptake was suppressed approximately equal to 90% by the intake of saturated triacylglycerols. As compared to polyunsaturated triacylglycerols, the intake of saturated lipids was also associated with significantly higher plasma LDL-cholesterol concentrations and lower levels of cholesteryl esters in the liver. PMID- 2989831 TI - Characterization of a continuous T-cell line susceptible to the cytopathic effects of the acquired immunodeficiency syndrome (AIDS)-associated retrovirus. AB - We have developed a continuous human T-cell line (A3.01) for the study of acquired immunodeficiency syndrome (AIDS)-associated retrovirus that mimics normal peripheral blood lymphocytes in susceptibility to viral cytopathic effect without the need for cell activation or conditioned medium. Following infection, substantial quantities of virus are produced during a 3- to 5-day period; the associated killing of cells can be monitored in a microtiter assay as a function of virus input. Southern blot hybridization of infected cellular DNAs indicated that no gross alteration occurred in the restriction maps of the proviral DNA during the transfer of virus to and its passage in A3.01 cells. This cell system offers an alternative to other AIDS retrovirus cell systems because it permits the monitoring of viral cytopathic effects. PMID- 2989832 TI - Transferrin receptors in rat brain: neuropeptide-like pattern and relationship to iron distribution. AB - We have characterized and visualized the binding of 125I-labeled transferrin to sections of rat brain. This saturable, reversible, high-affinity (Kd = 1 X 10(-9) M) binding site appears indistinguishable from transferrin receptors previously characterized in other tissues. Moreover, a monoclonal antibody raised to rat lymphocyte transferrin receptors could immunoprecipitate recovered intact transferrin solubilized from labeled brain slices, indicating that labeling was to the same molecular entity previously characterized as the transferrin receptor. The pattern of transferrin receptor distribution visualized in brain with both 125I-labeled transferrin and an anti-transferrin receptor monoclonal antibody are almost indistinguishable but differ from the pattern of iron distribution. Iron-rich brain areas generally receive neuronal projections from areas with abundant transferrin receptors, suggesting that iron may be transported neuronally. However, many brain areas with a high density of transferrin receptors appear unrelated to iron uptake and neuronal transport and form a receptor distribution pattern similar to that of other known neuropeptides. This "neuropeptide-like" distribution pattern suggests that transferrin may have neuromodulatory, perhaps behavioral, function in brain. PMID- 2989833 TI - Activity regulates the levels of acetylcholine receptor alpha-subunit mRNA in cultured chicken myotubes. AB - In vitro blocking the spontaneous activity of primary cultures of chicken embryo myotubes with tetrodotoxin increases approximately equal to 2-fold their content in surface acetylcholine receptor. To investigate this effect at the level of gene expression, chicken genomic DNA sequences coding for the acetylcholine receptor alpha subunit were isolated and characterized. They were shown to belong to a single-copy, polymorphic gene with at least two alleles in the chicken strain utilized. Probes derived from these genomic clones were used to quantitate levels of alpha-subunit mRNA. In culture, a 2-day exposure to tetrodotoxin increased these mRNA levels up to 13-fold, a value similar to that observed after denervation of chick leg muscle (approximately equal to 17-fold). Actin mRNA levels varied little in any of these experiments. These results support the notion that membrane electrical activity affects acetylcholine receptor expression by regulating the accumulation of the corresponding mRNAs. PMID- 2989834 TI - The human phenylalanine hydroxylase gene. PMID- 2989835 TI - The genomic sequencing technique. PMID- 2989836 TI - Restriction fragment length polymorphisms (RFLP's) and their use in mapping the human genome. PMID- 2989838 TI - Detection of mutations. PMID- 2989837 TI - DNA polymorphisms in collagen genes: potential use in the study of disease. PMID- 2989839 TI - Gene mapping and myotonic dystrophy. PMID- 2989840 TI - Molecular tools for the mapping of the human genome. AB - The rapid progress in molecular cloning and DNA analysis techniques, together with the use of cloned DNA probes from specific chromosomes of the human genome might allow localisation and ultimately identification of genes defined by single mutations. The discrepancy between genetic dimensions expressed in centimorgans, each corresponding to millions of base pairs and distances easily accessible by molecular techniques amounting to at the most hundreds of kilobase pairs may be bridged with some new cloning techniques partially developed at the European Molecular Biology Laboratory. These techniques were designed to allow rapid cloning and analysis of large regions of mammalian genomes. PMID- 2989841 TI - Welcome and objectives. PMID- 2989843 TI - Subclinical and clinical bluetongue disease in cattle: clinical, pathological and pathogenic considerations. AB - Calves immunized (sensitized) with alcide-inactivated bluetongue virus (BTV) developed immunoglobulin (Ig) E-specific antibody to BTV. Development of clinically apparent disease occurred after challenge with virulent BTV, and lesions correlated with peak levels of virus-specific IgE, an eosinophilic dermatitis, and high concentrations of histamine in the skin. IgE may be important in the pathogenesis of clinical bluetongue (BT) disease in cattle. PMID- 2989842 TI - Bluetongue and related orbiviruses. Proceedings of an international symposium. Monterey, California, January 16-20, 1984. PMID- 2989844 TI - Reproductive problems associated with bluetongue virus activity in Nebraska. AB - Abortions, stillborn calves, neonatal mortalities and excessive numbers of weak, slow-starting newborn have contributed to significant losses within the valuable breeding herds at the Roman L. Hruska U.S. Meat Animal Research Center. Conception rates of 70 to 80%, abortion storms and neonatal mortality of 5 to 25% have been observed. Vaccination programs control most of the common diseases that contribute to production losses. Management and nutrition are considered adequate. Excessive dystocia is thought to contribute to some of the excessive losses reported. Diagnostic efforts and epidemiological studies indicate that bluetongue virus (BTV) appears to be associated with some of the reproductive losses. Serotypes 11 and 13 have been isolated. Bovine virus diarrhea virus has also been isolated from weak calves that die at birth or shortly after birth. Serological studies reveal a periodically high incidence of bluetongue (BT) positive breeding animals. Collaborative efforts continue and must be expanded to further elicit the cause and economic impact of reproductive loss at the Meat Animal Research Center. PMID- 2989845 TI - Bluetongue in western Nebraska: an area herd study. PMID- 2989847 TI - Bluetongue and related orbiviruses: overview of the world situation. AB - BT is silently spreading in tropical and subtropical countries in the world. Serological surveys on BT in developing countries should be intensified and expanded. Research is needed to develop simplified virus typing techniques, to develop safe and effective vaccines and to establish procedures for differentiating BTV which produces clinical disease from that of low-virulency. It is considered necessary to develop a new disease reporting system which can accommodate both clinical and sub-clinical forms of BT. Knowledge on Ibaraki disease and EHD is limited. There is a need to intensify serological surveys in different parts of the world in order to grasp the global situation of these diseases and to plan strategies for them. To confine AHS in the south of the Sahara, the disease surveillance should be intensified in the Near East and northern Africa regions, and a stock of AHS vaccines for emergency use should be established. PMID- 2989848 TI - Salivary gland homogenates from the vector Culicoides variipennis may aid in detection of bluetongue virus in chronically infected cattle. AB - Studies were conducted on 2 cows chronically infected with bluetongue virus (BTV) acquired in utero from their dam. In previous research, BTV had been isolated 4 times from 1 cow and 8 times from the other. BTV was undetectable between spontaneous febrile and leukopenic episodes and antibodies to BTV were not detectable in the serum. Previous work had shown that bites by uninfected females of the vector Culicoides variipennis (Coquillett) caused virus to recrudesce in the blood to detectable levels in a bull with a similar BTV infection. After 1 of the 2 cows became hypersensitive to the bites of the vector, the cows were inoculated with homogenized salivary glands from the vector in an effort to increase the viral concentration to detectable levels. Bites by uninfected vectors were used 9.5-48 hr after inoculation of salivary glands in attempts to recover BTV biologically. In 2 of 6 experiments, after inoculations of the glands, BTV was isolated and confirmed from some of the blood samples and from some pools of C. variipennis females that had blood fed on both animals. PMID- 2989846 TI - Isolation of bluetongue virus from bull semen. AB - The efficacy of inoculation of Vero cell cultures or intravenous inoculation of chicken embryos in the isolation and titration of seminal bluetongue virus (BTV) was studied, as was the toxicity of bull semen for these 2 isolation systems. Frozen and thawed BTV-contaminated ejaculates collected during periods of viremia from 2 bulls experimentally infected with cell culture-adapted BTV serotype 17 were used in isolation, titration and fractionation studies. Blood collected from the 2 bulls concurrently with the semen was titrated in chicken embryos. Bull semen was toxic for both isolation systems. Toxicity was associated with both the spermatozoa and seminal plasma. Dilution of the semen at least 1:25, addition of peptone or tryptose broth to the diluent, limitation of adsorption time and postinoculation washing of cell culture monolayers all reduced the destructive effects of semen. Isolation of BTV was successful from 11 ejaculates and was titratable in 9 of these. Blind passage of surviving embryos or cell cultures at the endpoints of the titrations produced BTV isolations in 4 instances. The virus was never isolated from semen in the absence of concurrent viremia. Peak seminal BTV titers of 10(5.5) CEIVLD50/ml and 10(5.7) TCID50/ml were observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989849 TI - The current taxonomic status of the Culicoides vectors of bluetongue viruses. AB - In the regions where bluetongue (BT) viruses are known to occur the taxomonic literature is generally adequate for confident identification of the commoner species that are the most likely vectors. Unfortunately in each region there are difficult taxonomic problems in the accurate determination of some proven or potential vectors. These problems involve the so-called imicola and obsoletus groups of the subgenus Avaritia, the variipennis complex in the subgenus Monoculicoides and the schultzei group of the ill-defined subgenus Oecacta. In the imicola and schultzei groups sophisticated taxonomic studies now in progress have been successful in finding useful morphological as well as biological characters for recognition of the vector species. In the variipennis complex morphologically distinct eastern and western populations can be recognized, but their characteristics with respect to ease of colonization and ability to transmit BT virus (BTV) are not yet well known. The taxonomic status of the subspecies of variipennis known to breed in saline and alkaline habitats is still confused and badly in need of detailed study. PMID- 2989850 TI - Vectors of bluetongue virus in Australia. AB - Two of the 5 serotypes of bluetongue virus (BTV) known from Australia have been isolated from field collected insects. Serotype 20 was isolated in 1975 from a mixed pool of 214 insects containing several Culicoides species. Serotype 1 has been isolated from C. (Avaritia) fulvus Sen & Das Gupta collected at Beatrice Hill in the Northern Territory and from C. (Avaritia) brevitarsis Kieffer collected at Peachester in southeast Queensland. All other isolates of bluetongue (BT) group viruses have been made from sentinel cattle. An additional 2 species of the subgenus Avaritia, C. wadai Kitaoka and C. actoni Smith, 1 species of the subgenus Culicoides, C. peregrinus Kieffer and 1 species of the Schultzei group, C. oxystoma Kieffer have been infected in the laboratory. Serotype 20 was transmitted from sheep to sheep by C. fulvus and serotype 1 by C. fulvus and C. actoni. The infection rates established for Culicoides fed on sheep infected with serotype 20 were C. fulvus 62%; C. wadai 11%; C. actoni 2% and C. brevitarsis 0.3%. All species of insects successfully infected are widely distributed in the Oriental region. Attempts to infect species that are restricted in range to the Australasian region have been unsuccessful. The 3 species with highest experimental infection rates: C. fulvus, C. wadai and C. actoni, are confined in Australia to areas with an annual summer rainfall in excess of 800 mm, and do not penetrate to the drier areas where sheep are commercially husbanded. C. brevitarsis is the vector responsible for transmission in the coastal dairying areas, and although it does occur where sheep are reared, it is an inefficient vector. It breeds in discrete cow dung pats on pasture and is more closely associated with cattle than with sheep when the 2 hosts occur together. More than 1 species of Culicoides is responsible for the transmission of BT group viruses in Australia and no BT disease of sheep has been recorded because the more efficient vector species are not present in sheep rearing areas. PMID- 2989851 TI - Further data on the distribution of biting midges in southern Europe and the Mediterranean area, with special reference to Culicoides imicola. AB - Studies on the Culicoides midges of Spain and Turkey suggest that the range of C. imicola may extend to about 40 degrees N. However, catches of midges from mainland Greece, Sicily and Italy failed to reveal the presence of this species. These findings are discussed in the light of the possibility of the occurrence of bluetongue (BT) disease in these areas, consequent upon windborne spread of infected vectors from countries where the disease is endemic. PMID- 2989852 TI - Bluetongue virus isolation from pools of Culicoides spp in Israel during the years 1981 to 1983. AB - In 2 permanent suction light trapping stations at Bet-Dagan and Kabri designated to monitor bluetongue virus (BTV) activity, 12 isolations were made during 1981 to 1983 from C. imicola. Isolation attempts from C. obsoletus, which was dominant in Kabri and caught concurrently with C. imicola did not yield BTV, while isolations were made from C. imicola. Negative isolation atempts were also experienced in C. oxystoma, C. puncticollis and C. circumscriptus, while BTV was isolated from C. imicola. So far the following serotypes of BTV were isolated from Culicoides spp. in Israel: 2, 4, 6, 10 and 16. PMID- 2989853 TI - Orbiviruses from Culicoides in Florida. AB - Between 1980 and 1983, 45,484 Culicoides spp. collected in Florida near cattle have been examined for orbiviruses by attempted isolation in cell cultures and intravenous (IV) inoculations of embryonated chicken eggs. Bluetongue virus (BTV) serotype 2 was isolated from a pool of Culicoides insignis trapped at Ona. This is the 1st recovery of BTV from this species representing the 2nd new world species of Culicoides from which BT viruses have been isolated. C. insignis is a neotropical form which extends from northern Florida through the Caribbean region and much of South America. If this species is proven capable of biological transmission of BTV, it could help explain the distribution of antibodies to BTV in areas lacking C. variipennis. Epizootic hemorrhagic disease virus (EHDV) serotype 2 was recovered from several pools of C. variipennis collected at 2 beef cattle operations in Florida. No viruses were isolated from the small number of C. arboricola, C. stellifer and C. niger examined. PMID- 2989854 TI - Potential vectors of bluetongue and epizootic hemorrhagic disease viruses of cattle and white-tailed deer in Alabama. AB - Fourteen Culicoides spp. were collected on Holstein cattle in Alabama: C. arboricola, C. bickleyi, C. biguttatus, C. debilipalpis, C. guttipennis, C. haematopotus, C. obsoletus, C. paraensis, C. piliferus, C. sanguisuga, C. spinosus, C. stellifer, C. variipennis and C. venustus. Six Culicoides spp. were collected directly from white-tailed deer (Odocoileus virginianus): C. debilipalpis, C. niger, C. obsoletus, C. paraensis, C. sanguisuga and C. stellifer. Based on their host-feeding behavior, abundance and seasonal occurrence, the following 4 species warrant particular attention as potential vectors of bluetongue (BT) and epizootic hemorrhagic disease (EHD) viruses in the southeastern US: C. debilipalpis, C. obsoletus, C. paraensis and C. stellifer. PMID- 2989855 TI - Economic impact of bluetongue and related orbiviruses: Western Hemisphere. PMID- 2989856 TI - Observations on larval habitats of suspected Culicoides vectors of bluetongue virus in Florida. AB - Potential breeding sites for Culicoides variipennis (Coquillett) and C. insignis Lutz, suspected vectors of bluetongue virus (BTV) in Florida, were examined at 7 livestock facilities located in different geographic regions of the state. The 2 most productive habitats were mud contaminated by effluent from milking parlors (92.3 larvae/sample) and margins of vegetated ponds (18.8 larvae/sample). Four species of biting midges, C. crepuscularis Malloch, C. haematopotus Malloch, C. insignis, and C. variipennis were reared from immature stages extracted from substrate samples. Culicoides insignis was recovered from samples taken from muddy areas in pastures and margins of vegetated ponds, whereas C. variipennis was collected around waste lagoons and from mud contaminated by effluent from milking parlors. PMID- 2989857 TI - Methodology in preserving field-collected flies for bluetongue virus assay. AB - Preliminary tests were conducted to evaluate the use of phosphate buffered saline (PBS) as a collecting medium for the preservation of flies for virus assay from our bluetongue (BT) study areas in Colorado, USA. These tests were made possible by the use of a colony of Culicoides variipennis (Coquillett) that is highly susceptible to infection with BT virus (BTV). PMID- 2989858 TI - Oral infection of Culicoides (Diptera, Ceratopogonidae) with viral agents, using fine glass needles. AB - Oral infection of haematophagous insects with arboviruses to test their vector competence can be a difficult procedure in the laboratory since "wild caught" insects often prove reluctant to feed, once captured. This paper describes a technique using virus-charged glass needles to infect Culicoides spp. orally with bluetongue virus (BTV). C. variipennis, a known vector of BTV, was found to support virus multiplication with this technique, giving results comparable to those obtained using standard oral infection procedures. Other species of Culicoides that had previously been difficult or impossible to infect orally in the laboratory also engorged upon virus suspensions. The technique provides a means of assessing the vector competence of these and other similar Culicoides species. PMID- 2989859 TI - Laboratory infections of Culicoides debilipalpis and C. stellifer (Diptera: Ceratopogonidae) with bluetongue virus. AB - Six species of field-collected Culicoides females were used in preliminary laboratory tests: C. biguttatus, C. debilipalpis, C. obsoletus, C. stellifer, C. variipennis and C. venustus. Tests were conducted to determine if females would take a blood meal through a membrane, if blood fed females could be maintained in the laboratory for up to 14 days and if they could support replication of bluetongue virus (BTV) serotype 11. One pool of C. stellifer (23 specimens) assayed positive for BTV 7 days following intrathoracic inoculation, which datum suggests that this species might be capable of harboring the virus. One of 52 individually assayed C. debilipalpis females in 2 trials was positive for BTV at 14 days after membrane feeding on a mixture of BTV and sheep blood (infection rate 1.9%), and 1 of 2 pools of C. debilipalpis (26 specimens) was positive 14 days after blood feeding. These data for C. debilipalpis showed that the species can pick up and harbor BTV through replication of the virus. PMID- 2989860 TI - The virogenesis of bluetongue virus in Culicoides variipennis. AB - A direct immunofluorescence (IF) technique was developed to detect bluetongue virus (BTV) antigen in Culicoides variipennis. After approximately 10 days extrinsic incubation (EI), the IF test was as sensitive as virus isolation for detection of infected flies. The IF technique was used to determine the virogenesis of BTV in the fly. After 6 days EI, virus antigen was detected in most midguts; by 10 days, most head squash preparations contained antigen. Other secondary target organs, such as ovaries, abdominal ganglia and thoracic muscle, only infrequently contained detectable BTV antigen. PMID- 2989861 TI - Overview of the orbiviruses. PMID- 2989862 TI - Classification of orbiviruses: a need for supergroups of genera. AB - There has been concern that the present nomenclature system for the members of the Reoviridae family, and particularly the Orbivirus genus, does not represent the actual relationships exhibited between the members. In order to follow the conventions established by the International Committee for the Taxonomy of Viruses (ICTV), it is tentatively proposed that the present Reoviridae genera be upgraded in status to the following sub-families: reovirinae, orbivirinae, Fijivirinae, cypovirinae, rotavirinae, coltivirinae and phytoreovirinae. Below the sub-family level, divisions of genus (equivalent to superserogroup, serocomplex or supergroup), sub-genus (equivalent to serogroup), species (equivalent to serotype or virus), and variant (equivalent to sub-serotype or genotype) could be created. In the orbivirinae this would result in 2 genera of cyanovirus (bluetongue, epizootic hemorrhagic disease, Eubenangee and Palyam sub genera) and Kemerovovirus (Chenunda, Great Island, Kemerovo and Wad Medani sub genera) and a number of ungrouped sub-genera (African horsesickness, Changuinola, Corriparta, equine encephalosis, Wallal and Warrego sub-genera and the remaining ungrouped viruses). It is hoped that further biochemical studies shall confirm these groupings at a more fundamental level and eventually a system recognising the double-stranded RNA gene product relationships shall evolve. PMID- 2989863 TI - Bluetongue, epizootic haemorrhagic disease of deer and related viruses: current situation in Australia. AB - Since 1975 3 serotypes of bluetongue (BT) virus (BTV) have been identified in Australia: BTV1 (CSIRO156), BTV20 (CSIRO19) and BTV21. At present 2 further BT viruses (DPP90 and DPP192) have been isolated from the blood of healthy cattle in the Northern Territory (NT) and are undergoing identification. There is serological evidence for BTV15 infection in Western Australia (WA) and the NT, and a background level of serological activity to BTV serotypes 1 to 17. In addition, over 50 isolations of epizootic hemorrhagic disease of deer (EHD) viruses (EHDV) have been made and can be divided into 5 main serotypes, CSIRO157, CSIRO439, CSIRO753, CSIRO775 and DPP59. All 5 serotypes occur within the ecological range of Culicoides brevitarsis, however, as for BTV, the high prevalence of antibodies in cattle in the Kimberley region of WA and the top end of the NT suggests other vectors may also be responsible. CSIRO439 has been shown to be closely related serologically to Ibaraki virus while the other EHD viruses were distinct from Ibaraki and from each other. The complex serological relationships between BTV, EHDV and Palyam virus groups are currently being examined using serological and biochemical techniques. It is often difficult to determine the differences between BTV and EHDV serogroup members. Antibodies to both BTV and EHDV have been found in cattle, buffaloes and deer but there has been no evidence of clinical disease in any of these species or in sheep under field conditions in Australia. PMID- 2989865 TI - An electron microscopic study of blood cells from calves experimentally infected with bluetongue virus. AB - Cellular elements from blood samples of calves experimentally inoculated with a quadrivalent mixture of bluetongue virus (BTV) types 10, 11, 13 and 17 were examined by transmission electron microscopy (TEM). Virus-like particles were observed within cytoplasmic vacuoles of infected agranular leukocytes from inoculated calves on postinoculation day (PID) 14. No virus-like particles were seen associated with other cellular blood elements nor were they observed in blood samples obtained prior to virus inoculation. The intravacuolar viral particles were 60nm in diameter, had a cockleburr appearance and lacked the outer polypeptide coats of mature virions. Increased cytoplasmic vacuolation was the most noticeable change in the infected cells. BTV infection was confirmed by viral isolation and serological testing. Pyrexia was the only consistent clinical sign seen in the viremic calves. PMID- 2989866 TI - Morphology of bluetongue virus-infected Aedes albopictus (C6/36) cell culture. AB - Morphological changes observed in bluetongue virus (BTV)-infected Aedes albopictus (C6/36) suspension cell culture at 28 C were compared to those observed in mammalian stationary cell culture at 37 C. The presence of cytoplasmic macrotubules, viroplasms and progeny virions was confirmed and appears to be the same as seen in mammalian cell culture. In addition progeny virions were observed budding from cell membranes where they appeared to acquire a lipoprotein envelope. Intranuclear macrotubules that are the same diameter as the cytoplasmic macrotubules were also seen. C6/36 cell culture is a model for gaining insight into BTV-replication in the insect host. PMID- 2989864 TI - Speciation in orbiviruses. AB - The definition of Orbivirus species should be based on the ability of virus populations to reassort genetic information. Application of the definition of biological species to orbiviruses enables consideration to be given the evolutionary tendencies of virus populations and to mechanisms for generating diversity within orbiviruses. PMID- 2989868 TI - Isolation and propagation of bluetongue virus in embryonating chicken eggs. AB - The incidence of bluetongue (BT) disease in sheep in Israel during 1964-1982 was presented. The intravenous (IV) and the yolk-sac (YS) routes of inoculating embryonating chicken eggs (ECE) for primary isolation and propagation of BT virus (BTV) were compared and assessed. It was shown that the IV route of inoculation was about 100- to 1,000-fold more sensitive than the YS route; also, by the IV route, virus isolation was more rapid and assays were more clear cut. About 30% of virus isolations from field samples obtained by the IV route were missed when the YS route was used. Ten to 13-day-old ECE were equally susceptible to replication of BTV. The growth characteristics in ECE were different for BTV serotypes 2, 4, 6, 10 and 16. Titratable virus replication preceded ECE mortality by 1 or 2 days. The maximum viral titre in live ECE was usually lower than the maximum titre in dead ECE. BTV serotypes 2, 4, 6 and 16 were isolated from Culicoides imicola in IV inoculated ECE. PMID- 2989867 TI - The search for bluetongue viruses in Australia. AB - In the 200 years that cattle and sheep have been present in Australia no evidence of bluetongue (BT) disease has ever been reported. The discovery in 1977 that a virus isolated from Culicoides species collected in 1975 was bluetongue virus (BTV) triggered a search for BT viruses and their vectors throughout the country. Between 1975 and 1983, 46 strains of 5 serotypes of BTV were isolated by various workers from the blood of subclinically infected sentinel cattle or from Culicoides species. The serotypes are 1, 20, 21 and 2 others not yet fully characterized. By testing stored sera from cattle and buffaloes, antibodies to serotype 21 were found in sera collected in 1958, but antibodies to serotypes 20 and 1 were not found earlier than 1973 and 1969 respectively. Discrepancies between the results of BTV group antibody and neutralizing antibody surveys indicate that other serotypes may yet be found. Neutralizing antibodies to BTV have been found in cattle, buffaloes, deer, goats and sheep, but mammals indigenous to Australia are apparently not being infected. In the course of the search, Ibaraki virus and 4 other viruses of the epizootic haemorrhagic disease (EHD) group were found. Infection with EHD viruses has produced heterotypic antibodies detected in tests for BT group antibodies. The prevalence of antibodies to BT and EHD viruses in cattle is greater in the northern 1/3 of the continent than the middle 1/3 while the southernmost 1/3 is free. By the sequential sampling of sentinel cattle it has been determined that infection with BT and EHD viruses occurs mainly in the latter part of summer, and autumn. The seasons of the year, and the locations from which BT and EHD viruses originated were consistent with the serological findings. Isolations of BT and EHD viruses have also been made from insect vectors, BTV serotype 1 from C. fulvus and C. brevitarsis, plus serotype 20 from a mixed pool of Culicoides species and 2 different EHD viruses from C. brevitarsis. The susceptibility of other Culicoides species to experimental infection with BT and EHD viruses suggests that they may be involved in the natural spread of these viruses. BT and EHD viruses appear to be non-pathogenic for cattle under experimental conditions, but mildly or moderately pathogenic for sheep. The clinical signs produced are consistent with those produced experimentally in other countries. PMID- 2989869 TI - Isolation and identification of bluetongue virus: a serotype new to the U.S. AB - There are 5 known serotypes of bluetongue (BT) virus (BTV) in the US: 2, 10, 11, 13 and 17. The most recently discovered US serotype, BTV 2, was isolated from blood collected from cattle at Ona, Florida in 1982. Isolations were made from the September through November bleedings and from 1 pool of Culicoides insignis Lutz collected in October. Seventeen viral isolates were obtained by injecting the samples into embryonated chicken eggs (ECE) followed by serial passage onto BHK-21 cells. A single isolate was made from inoculation of pooled bovine blood into sheep. The isolates were shown to be Orbivirus-like particles by electron microscopy and were identified as BTV by indirect immunofluorescence tests. All the isolates were found to be serotype 2, however genome analysis using polyacrylamide gel electrophoresis (PAGE) identified 2 distinct electropherotypes. There were major differences between the 2 electropherotypes at least in genome segments 1, 5, 7, 8 and 9. The electropherotypes of viral isolates from bovine blood collected in September, 5 of those from the October bleedings, the insect isolate, and that from pooled blood were identical by PAGE and were designated Ona A. This electropherotype was also found to be indistinguishable from the prototype of the African serotype 2. The 2nd electropherotype, Ona B, included 3 of the viral isolates from the October bleedings and the single isolate from November. Ona A appeared to be closely related to the African prototype while Ona B may have resulted as a variant of Ona A after being subjected to a different environment. Both electropherotypes, however, appear to be stable forms of BTV serotype 2. PMID- 2989870 TI - Biochemical characterisation of Australian orbiviruses. AB - There are over 30 antigenically distinct orbiviruses found in Australia, including members in the bluetongue virus (BTV) and epizootic hemorrhagic disease of deer virus (EHDV) serogroups. Genomic RNA profiles were analysed by polyacrylamide gel electrophoresis (PAGE) on both 10% Laemmli and tris-borate EDTA-(TBE)-urea gels. There was considerably more variation in the RNA profiles in Laemmli gels than was apparent in the TBE-urea gels. Since the latter system separates on molecular size, then presumably migration in the Laemmli gels may depend upon molecular weight (MW) and conformation. Analyses of 35S-methionine labeled proteins in virus-infected cells was carried out by PAGE in 10 to 20% gradient Laemmli gels. Twelve to 15 virus-specific labeled protein bands were observed in cells infected with orbiviruses. A detailed analysis of Australian BTV 1 isolates was made to identify these proteins. In addition to previously reported proteins (P1 to P8A) an additional low MW protein, P9, was observed (approx. MW 12,000). In the EHDV serogroup, 3 viruses (Ibaraki, CSIRo402 and CSIRo439), which were very closely related by virus neutralization tests and in protein-PAGE, were distinct in their migration by RNA-PAGE. Analyses of the individual RNA segments by 1-dimensional T1-ribonuclease oligonucleotide mapping showed minor differences between Ibaraki and the Australian EHDV isolates, suggesting that the 3 viruses have similar 3'-terminal RNA sequences. These studies suggest that Ibaraki (IBA) virus is closely related to but distinct from the Australian isolates. PMID- 2989871 TI - Immune response against the purified serotype specific antigen of bluetongue virus and initial attempts to clone the gene that codes for the synthesis of this protein. AB - Sheep were injected with different amounts of purified protein P2 of bluetongue (BT) virus (BTV). About 3 X 50 mcg was required for the induction of neutralizing antibodies. Sheep injected with 3 X 10 mcg were, however, still largely protected when challenged with virulent virus. This has suggested the possibility of using P2 as a subunit vaccine and initiated an investigation of the possibility of synthesizing P2 by DNA-recombinant technology. In order to clone the gene that codes for the synthesis of P2 both the "shotgun" approach with unfractionated dsRNA and cloning of isolated segment 2 were investigated. The basic approach was to convert the dsRNA to DNA which was cloned into the Pst 1 site of E. coli plasmid pBR322. The largest BTV-specific insert that was obtained in the initial experiments was just more than 2,000 base pairs long. The largest insert obtained when isolated segment 2 dsRNA was cloned was about 1,200 base pairs which represents about 1/3 of the P2 gene. PMID- 2989872 TI - Molecular cloning and hybridization studies on bluetongue virus serotype 17. AB - The dsRNA of bluetongue virus (BTV) serotype 17 has been reverse transcribed and dsDNA copies of the viral RNA have been cloned into the plasmid vector pBR-322. Segments ranging from 3 kilobases to less than 500 bases have been cloned and at present 1 of the clones has been identified by hybridization to the genome segment of its origin (genome segment 7). Identification of further clones is proceeding. The techniques for northern blotting BTV dsRNA onto 2 aminophenylthioether (APT) paper and the detection of the transferred RNA by 32P labelled dsRNA or cDNA probes have been standardized. Cross-hybridization studies can be used to detect genetic relationships of different serotypes and isolates of BTV. PMID- 2989873 TI - Cloning and nucleotide sequencing of bluetongue virus genomes. AB - This is the 1st report of molecular cloning of bluetongue virus (BTV) gene using DNA recombinant techniques. A number of complete and overlapping clones of BTV genes have been cloned into pBR322 by standard procedures with some modification. The full length clones were confirmed by matching the terminal cDNA nucleotide sequences with those previously determined for the termini of double-stranded genomic RNA (Rao et al., 1983). The complete sequence of the cDNA clone of RNA segment 3 of US serotype 17 as determined by the method of Maxam and Gilbert (1980) has been presented here. The clone is 2,772 nucleotides (1.78 X 10(6) daltons) long excluding the 3' polyadenylic acid sequence and has an open reading frame which encodes a protein of some 901 amino acids (103,412 daltons). PMID- 2989874 TI - Bluetongue in the United States. AB - The virus of bluetongue (BT) was 1st isolated and identified in the US in 1952 from sheep in California. A disease in sheep in Texas (soremuzzle) was observed in 1947, reported in 1952 and similarities to BT were discussed. It is possible BT existed in Texas for several years prior to that time. There are 23 immunologic serotypes of BT, and 5 are known to occur in the US. These are types 10, 11, 13, 17 and 2, the latter having been recognized only in 1983 in cattle in Florida. In the US, BT was 1st recognized as a disease of sheep, but by the 1960s BT was known to occur in cattle with clinical manifestations which could be mistaken for vesicular stomatitis, infectious bovine rhinotracheitis and foot-and mouth disease. Epizootic hemorrhagic disease (EHD) of deer, caused by orbiviruses related serologically to BT, was 1st recognized in New Jersey in 1956. Two serotypes of EHD virus have been identified in the US. PMID- 2989875 TI - Analysis of the terminal sequences of the genome segments of four orbiviruses. AB - The RNA sequences of the terminal regions of the genome segments of 4 different orbiviruses were analysed. In 3 of these [bluetongue virus (BTV) types 1 and 20 and Ibaraki virus, a member of the epizootic hemorrhagic disease (EHD) serogroup], 1 strand of all of the genome segments analysed ends in 3'CAAUUU...5' while the other strand ends in 3'CAUUCA...5'. These conserved sequences are identical to those reported for BTV types 10 and 11. The 3' terminal sequences of segments 3 and 10 of the BTV 1 genome (including the conserved regions) were confirmed by the detection of exactly complementary sequences at the 5' termini of the ssRNA strands of opposite polarity. This also confirmed for these dsRNA segments (and by analogy for all of the genome segments of these viruses) that the dsRNA molecules are fully base-paired end to end. Using in vitro synthesised mRNA of BTV 1 in annealing experiments with the separated ssRNAs from the individual genome segments, it was shown that in each case the strand ending in 3'CAUUCA...5' is of the same polarity as the mRNA (+ve), while the strand ending in 3'CAAUUU...5' is of the opposite (-ve) polarity. The 4th virus analysed (Tilligerry, a member of the Eubenangee serogroup) only had 5 conserved bases at the 3' termini of 1 strand of each genome segment (3'CAU-CA...5') and 3 conserved bases at the 3' termini of the other strand (3'CA--U...5'). Considerable sequence homology was found in the near-terminal nonconserved regions of comparable genome segments, particularly between the different BTV types. There was little evidence however for absolute conservation of "segment specific" sequences in these regions of the RNA, as reported for rotaviruses. The percentage of sequence homology found in the terminal regions of the genome segments between members of the BTV, EHD and Eubenangee serogroups examined correlates with the serological interrelationships between these groups of viruses. The conserved terminal sequences and sequence homology in these regions of the RNA are discussed as to their role in virus replication and their significance to the reassortment of genome segments between different viruses during dual infections. PMID- 2989876 TI - Genome RNAS of virulent and attenuated strains of bluetongue virus serotypes 10, 11, 13 and 17. AB - An improved system was developed for the electrophoretic resolution of bluetongue virus (BTV) genome segments. This procedure was used to compare the genome segments of wild type (W) and vaccine (V) strains of BTV serotypes 10, 11, 13 and 17. Numerous alterations in the migration of individual genome segments could be detected between W and V strains of each serotype, suggesting that many mutations had occurred during attenuation of the V strains. In addition, electrophoretic polymorphisms not previously detected, were found between W strains of the 4 serotypes. PMID- 2989877 TI - Immunochemical analyses of Australian bluetongue virus serotypes using monoclonal antibodies. AB - In order to characterize the immunochemical role of bluetongue virus (BTV) specified proteins and provide reagents capable of defining the serological relatedness of bluetongue (BT) serotypes and their relationship with other orbiviruses, a panel of 16 IgG monoclonal antibodies was raised to the Australian BTV serotypes, isolate CSIRO156 (BTV 1), CSIRO19 (BTV20) and CSIRO154 (BTV21). Analyses of virus-coded polypeptide specificities of these monoclonals using enzyme-linked immunosorbent assay (ELISA), a radioimmunoprecipitation assay (RIPA), and a virus neutralization assay, revealed the outer coat viral protein P2 to have a major role in the neutralization of both CSIRO156 and CSIRO19. Presumptive evidence for the involvement of the P3 protein in the neutralization of CSIRO19 was also obtained. The virus-specified non-structural protein P6A induced a group reactive immune response to all 3 serotypes. Antigenic relationships between P3 of CSIRO156 and P2 of CSIRO19 were found, and an analysis of the relationships between epitopic regions on P2 and P3 of both viruses revealed several distinct immunogenic sites exist on the P2 protein. PMID- 2989878 TI - Growth characteristics of virulent and attenuated strains of bluetongue virus serotypes 10, 11, 13 and 17. AB - The growth properties of wild type (WT) and live, attenuated vaccine (V) strains of bluetongue virus (BTV) serotypes 10, 11, 13 and 17 were compared. All BTV strains produced maximal yields when grown at 34 C and harvested 24-36 hr after infection. The 4 V strains were temperature-sensitive (ts) for growth at 39 C as compared to the corresponding WT strains. The 4 V strains were ts for plaque formation when 37 C was used as the nonpermissive temperature. One-step growth curves showed that only the V strain of type 13 had significantly altered growth kinetics as compared to the corresponding WT. All BTV strains were strongly cell associated throughout the growth cycle. PMID- 2989879 TI - Gamma ray sensitivity of bluetongue, EHD and African horsesickness viruses and their precipitating and complement fixing antigens. PMID- 2989880 TI - Role of the immune system in bluetongue host-viral interactions. AB - Immune responses involving both B and T lymphocyte subpopulations have been demonstrated in sheep and cattle. The immune responses appear to be associated with the modulation of disease expression in sheep and cattle. An immature immune system in fetal lambs and calves permits virus to replicate with little or no host interference. Viral teratogenicity may consequently lead to malformed newborns, fetal deaths, abortion or readsorption. Adult sheep and cattle appear to respond somewhat differently to bluetongue virus (BTV). In both species T cell or cell mediated (CMI) and B cell or humoral mediated immunity (HMI) occur. There is a correlation between CMI response and protective immunity in sheep. Although these responses occur in cattle it appears that an IgE mediated hypersensitivity leads to expression of clinical disease and suppressive modulation of protective immunity. PMID- 2989882 TI - The position of the European community in relation to bluetongue. AB - The current position in the European Community concerning bluetongue (BT) is briefly explained. Mention is made of legislation covering this disease and the application of such legislation within the Community. PMID- 2989881 TI - An overview of diagnostics for bluetongue. PMID- 2989883 TI - Bluetongue: diagnostic/antigenic interpretation. PMID- 2989884 TI - Problems in the interpretation of diagnostic tests due to cross-reactions between orbiviruses and broad serological responses in animals. AB - Tests presently used for the diagnosis of infections by bluetongue virus (BTV) or related orbiviruses are based on the use of 2 types of serological reactions. Those that are considered group-reactive tests are the agar gel diffusion precipitin (AGDP), complement-fixation (CF) and fluorescent antibody tests and those that are considered type-specific are a wide variety of virus neutralization tests (50% and 80% plaque reduction, plaque inhibition and microtiter neutralization) and cross-protection tests. These tests suffer from problems of standardization between laboratories and of specificity. Group reactive tests (AGDP and CF) for the BTV serogroup also detect cross-reactions with viruses in the epizootic hemorrhagic disease virus (EHDV), Eubenangee (EUB) and Palyam (PAL) serogroups, with the EHDV cross-reactions being of particular concern. Further, multiple infections of cattle with PAL serogroup members can produce antibodies which will react to BTV and EHDV serogroup antigens in serological tests. Multiple infections of animals with related viruses can produce antibodies which will cross-react with orbiviruses in type-specific, virus neutralization tests to a virus which the animal has not previously been exposed. These observations stress the need to evaluate the tests at present being used, to assess the risks of cross-reactions between related orbiviruses and to develop new tests of defined specificity. PMID- 2989885 TI - Bluetongue: a review of the immune response of sheep and cattle to bluetongue virus infection. PMID- 2989886 TI - The use of serology in bluetongue epidemiology. PMID- 2989887 TI - Bluetongue and related orbivirus diagnosis in the United States. AB - The serologic test for bluetongue (BT) that was used in the US from 1968 to 1980 to qualify animals for export was the modified direct complement-fixation (MDCF) test. The MDCF test was replaced by the immunodiffusion (ID) test in 1980. In January 1984, there were 70 laboratories approved by the USDA to conduct BT ID export testing. Both tests are used at the National Veterinary Services Laboratories (NVSL) for the serologic diagnosis of suspected cases of BT and/or epizootic hemorrhagic disease (EHD). Serologic surveys were conducted on bovine samples collected from 18 northeastern states in the winters of 1977/1978, 1979/1980, 1982/1983 and 1983/1984. The percents of positive animals were 0.3, 0.3, 5.4 and 0.9. In 1977/1978 samples were collected from all states; 18.5% were positive. The primary technique used for the isolation of BT virus (BTV) at the NVSL is intravenous inoculation of embryonating chicken eggs with subpassage into Vero-M cell line cultures. Baby hamster kidney (BHK) cell line cultures are used for isolation of EHD virus (EHDV). Each virus is identified by staining with specific fluorescent antibody conjugate. PMID- 2989888 TI - An overview of Colorado tick fever. AB - Certain features of Colorado tick fever (CTF) virus and the disease it causes may be relevant to studies on bluetongue virus (BTV), or other orbiviruses. Rapid and easy detection of viral antigen in infected tissues and peripheral blood cells by immunofluorescence staining facilitate diagnosis of the disease. The prolonged (3 4 months) viremia is due to persistent intracellular infection, particularly of erythrocytes, in which the virus is protected from antibody or other host defense mechanisms. This results in more efficient maintenance of the virus cycle in nature, but might lead to adverse effects in the human host. Clues to understanding chronic viral infections or viral immunosuppression might be gained by further research on CTF and other orbiviruses. PMID- 2989889 TI - Importance of ovine cytotoxic T cells in protection against bluetongue virus infection. AB - In sheep, bluetongue virus (BTV) was shown to induce anti-BTV cytotoxic T lymphocytes (CTL) and their effect to be maximal around 14 days post inoculation (p.i.) of virus. Using cellular adoptive transfer techniques in monozygotic sheep, such cells were shown to partially protect animals from BTV challenge. A short-lived cross-protective mechanism was identified involving thoracic duct lymphocytes (TDL) and nonneutralising antibody. These observations suggest that T lymphocytes play an important role in protection against BTV and that current vaccine design based on in vitro serological typing of BTV can be improved. PMID- 2989890 TI - Humoral and cellular immune response of sheep to bluetongue virus. AB - Plaque cloned strains of the 4 US bluetongue (BT) virus (BTV) serotypes (10, 11, 13 and 17) were pathogenic to sheep and induced mild clinical responses. The clinical responses coincided with the highest titer of viremia reached by day 7 following primary infection. The 4 BTV strains were immunogenic, inducing group specific (precipitating) and type-specific (neutralizing) antibodies. Primary infection induced an immune response which protected the animals against secondary challenge with the homologous virus. Peripheral blood lymphocytes (PBL) obtained from infected animals responded specifically to in vitro stimulation with pure BT viral antigens. PBL responded to both homologous and heterologous BTV antigens indicating a cross-reactive nature of the lymphocyte response. Perturbations were observed in PBL response to in vitro stimulation with the mitogens phytohemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM). The response to mitogens was depressed transiently following primary infection and secondary challenge. A significant increase was reached by 3 weeks following primary infection and then gradually leveled off to normal. The specific stimulation of PBL in response to viral antigens and the increase in response to mitogens, as in vitro correlates of cell mediated immunity (CMI), were suggested to play a role in the clearance of BTV infection. Immunoblotting was applied to characterize the specificity of the serologic response to BTV. Sheep antisera to BTV detected 11 specific viral proteins. The maximum response on day 28 post inoculation (PI) detected 11 proteins including both the major and minor protein components.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989891 TI - Monoclonal antibodies and bluetongue virus diagnosis. AB - A blocking enzyme linked immunosorbent assay (ELISA) is described for the detection of group specific antibodies to bluetongue virus (BTV). The test is based upon the interruption of the reaction between BTV antigen and a group specific monoclonal antibody raised against BTV by the addition of serial dilutions of either bovine or ovine test sera. The presence of group specific antibodies to BTV in the test serum, results in inhibition of the monoclonal antibody which is detected as a reduction in the expected optical density (OD) reading. The test is capable of the detection of antibodies to all 22 types of BTV but unlike the agar gel precipitin (AGP) test does not show cross-reactions with antibodies to epizootic haemorragic disease (EHD) viruses. Antibodies to cellular proteins which give rise to complications in the AGP test and indirect ELISA are not detected by this assay. Preliminary studies indicate that due to its high specificity and sensitivity, the blocking ELISA is an ideal alternative to the AGP test. The use of such a monoclonal antibody would also facilitate the standardization of diagnostic testing between laboratories. PMID- 2989892 TI - Welcome and objectives. PMID- 2989893 TI - Monoclonal antibodies raised against bluetongue virus detect viral antigen in infected tissues using an indirect immunoperoxidase method. AB - An indirect immunoperoxidase method was developed to determine the presence of viral antigen in bluetongue virus (BTV) infected tissues. Embryonating chicken eggs were infected with each of the 4 United States BTV serotypes (10, 11, 13, 17) and the chorioallantoic membranes were subsequently formalin fixed and embedded in paraffin. The infected membranes were examined for the expression of viral antigen. Monoclonal antibodies raised against BTV 17 (BT 21A and BT 98A) were used as the primary antisera in the immunoperoxidase procedure. Excellent specific staining was observed when each of the antisera was applied to membranes expressing antigens of homologous BTV serotype. BT 98A showed outstanding reactivity with membranes infected with heterologous BTV serotypes as well. PMID- 2989895 TI - Is the AGID test group-specific for epizootic haemorrhagic disease virus? PMID- 2989894 TI - Plaque neutralization cross reactivity of post infection bovine sera amongst bluetongue virus serotypes. PMID- 2989896 TI - Epidemiology of orbiviruses. PMID- 2989897 TI - Epidemiology of bluetongue in Australia: the vertebrate hosts. AB - Ruminants became established in Australia after European settlement which commenced in the late 18th century. Bluetongue (BT) disease has never been suspected in cattle or sheep in this country. The species of ruminants found to have antibodies to BT and epizootic haemorrhagic disease (EHD) viruses are cattle, buffaloes, deer, goats and sheep. No antibodies have been found in pigs, horses, marsupials (kangaroos and wallabies) or humans, even in areas of high prevalence of BT and EHD infection. The serological and historical evidence suggests that Indonesia, where ruminants have been indigenous for a very long period, is a more likely source of infection for the Australian BT and EHD viruses than Papua-New Guinea though the method of their entry to Australia is unknown. It is unlikely that all the BT viruses entered from there in ruminants as there has been no importation of ruminants from Indonesia this century. Antibody surveys of cattle have shown that 2 of the 5 Australian BTV serotypes, DPP90 and DPP192 (Gard GP: unpublished data), are restricted to a limited area of the northern part of the Northern Territory of Australia, 1 (serotype 20) to the north of the 18th parallel of south latitude and the remaining 2 (serotypes 1, 21) to the north of the southern limit of the distribution of the suspected vector Culicoides brevitarsis. The prevalence of antibodies to BT and EHD viruses is highest in the far north of the Australian continent and declines rapidly in the middle 3rd. In the southern 3rd, the cattle and sheep are free of BT and EHD antibodies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989898 TI - Bluetongue epidemiology in the Middle East. PMID- 2989899 TI - Epidemiology of two orbiviruses in California's native wild ruminants: preliminary report. AB - Between 1978 and 1983 we collected more than 1,500 serum samples from California's native black-tailed deer (Odocoileus hemionus columbianus), 4 races of mule deer (O. h. sp.), tule elk (Cervus elaphus nannodes), Roosevelt elk (C. e. roosevelti), pronghorn antelope (Antilocapra americana), California bighorn sheep (Ovis canadensis californiana), Peninsular bighorn sheep (O. c. cremnobates) and desert bighorn sheep (O. c. nelsoni) and analyzed them for agar gel precipitating (AGP) antibodies to bluetongue (BT) virus (BTV). Various serologic tests for antibodies to a closely-related Orbivirus, epizootic hemorrhagic disease (EHD) virus (EHDV) of deer, were conducted on many herds in most years. Parallel attempts at virus isolation yielded 4 BT serotypes: 10, 11, 13 and 17. EHDV was isolated only twice. The herds sampled were representative of those found in various habitats under various management stages in California. Sampling schemes varied as attempts were made to pinpoint temporal, locational, weather, temperature--related and livestock grazing pressure factors influencing rates of infection. Some disease syndromes associated with isolated BTV and BT antibodies are discussed. This work is the 1st attempt to quantitate the level of Orbivirus infection in wildlife in California. PMID- 2989900 TI - Serotypes of bluetongue virus present in Saudi Arabia. AB - The widespread occurrence of infection with bluetongue virus (BTV) in Saudi Arabia has been demonstrated by immunodiffusion testing. Subsequently, 31 sheep sera and 1 goat serum with clear precipitating activity were examined by neutralization test using the 22 reference serotypes of BTV. Only 12 sheep sera neutralized 1 or more of BTV serotypes: 6 (1 serum), 14 (2 sera), 17 (4 sera), 19 (1 serum) and 20 (11 sera). The goat serum neutralized only serotype 19 of BTV. On the other hand, 19 sheep sera failed to neutralize any known BTV serotype. The significance of these serological findings was discussed and planning for further studies to isolate local BTV strains from sentinel flocks of lambs and calves was proposed. PMID- 2989901 TI - Bluetongue in Cyprus. PMID- 2989902 TI - Bluetongue: virological and epidemiological observations in Israel. AB - Bluetongue (BT) has become endemic in Israel. Five BT virus (BTV) types have been identified, of which BTV 4 is dominant. Results of the regular monitoring of viral activity in ruminants and Culicoides, including country-wide serological surveillance in bovines and virus recovery trials from bovine semen, are represented. The seasonal and geographical distribution, susceptibility of various species and applied preventive measures are discussed. Further studies are needed to establish the interseasonal epizootiology of BT in Israel. PMID- 2989904 TI - Bluetongue virus infection in Costa Rican and Colombian cattle. AB - Bluetongue virus (BTV) group antibodies are widely distributed in Costa Rica and Northern Colombia; prevalence is highest at lowest altitudes. Clinical evidence of bluetongue (BT) infection in cattle is not seen. Evidence exists of the circulation of BTV serotypes 6 and 14 in Costa Rica and BTV serotype 12, 14 and 17 in Northern Colombia in the period 1981-1983. Culicoides insignis is implicated as a probable vector in Colombia. PMID- 2989903 TI - First report of bluetongue antibody in Chile. AB - A sample of 1,752 bovine sera collected from 99 herds throughout the Tenth Region, Chile, from March 8 to November 20, 1982, was analyzed for bluetongue (BT) antibodies using the immunodiffusion (ID) test. The prevalence of BT virus (BTV) antibody was 19.6%. Sixty-four (64.6%) herds showed at least 1 positive animal, which suggests an important antibody distribution in the area. The examination of 500 sera of sheep in 14 herds indicated the following results: 8/500 animals were positive in 5/14 of the herds. PMID- 2989905 TI - Serological observations on the epidemiology of bluetongue virus infections in the Caribbean and Florida. AB - Serological surveys of cattle, sheep and goats have confirmed that infection with bluetongue virus (BTV) is common in Florida, Puerto Rico and St. Croix in the USA, in the Caribbean countries of Jamaica, St. Kitts/Nevis, Antigua, St. Lucia, Barbados, Grenada, Trinidad and Tobago, and the Bahamas, and in the South American countries of Guyana and Suriname. In most countries, over 50% of ruminant livestock have antibody to BTV as assessed by the bluetongue immunodiffusion (BTID) test. A sentinel animal system operating in Florida and 4 islands in the Caribbean has established that the transmission of BTV is seasonal, with most animals becoming infected in late summer and fall. In Florida, it appears that there may be some years when little virus transmission occurs among cattle. Examination of sera from yearling animals and sentinels in the region for antibody to the range of serotypes of BTV recognized worldwide, has resulted in a) the isolation of BTV type 2 from cattle in Florida - the 1st time this virus has been identified in the Western Hemisphere - and b) the recognition that the range of serotypes of BTV present in the Caribbean may be different from those in the USA. No clinical disease has been associated with BTV during the period of these studies (1979-83). PMID- 2989907 TI - Epidemiologic study of bluetongue virus on the Tejon Ranch, California: vector, host, virus relationships. AB - Culicoides gnats were monitored from April through November, 1981, on the Tejon Ranch, Kern County. Levels of gnat breeding were determined by quantitative sampling of substrate from intermittent and permanent water sources. Adult populations were measured by light trap collections taken around pens holding sentinel beef cattle, dairy calves, sheep, goats and deer. Adult gnats were collected in the environs of the study area, and blood samples were obtained from sentinel animals for laboratory diagnosis and isolation of bluetongue virus (BTV). C. variipennis was 1st observed in various water sources in April, with a continuous increase in density through October. Adult populations reflected this trend in density through October. Both immature and adult gnat populations rapidly decreased in November. BTV serotypes 11, 17 and 13 were isolated from C. variipennis from August through October. BTV serotypes 11 and 17 were initially isolated from nearly all sentinel sheep, goats and dairy calves from late July through September, whereas isolates of 11 and 17 were made from only 2 beef cows and 2 deer in August and September. Sheep were the only sentinel animals that showed clinical signs of BT disease. PMID- 2989906 TI - Epizootiological study of bluetongue virus infection in California livestock: an overview. AB - An epidemiologic program was undertaken in California to study bluetongue virus (BTV) infection in domestic livestock. The study was designed to determine: a) prevalence of BTV infection, b) serotypes of BTV actively causing infection, c) seasonality of infection and d) species infected. A total of 8,751 cattle, 14,639 sheep and 4,785 goats were tested over the 3 1/2 year study. Serologically, 41% of the cattle, 42% of the sheep and 21% of the goats were positive. Virologically, 2.4% of the cattle, 1.4% of the sheep and 0.7% of the goats were viremic. One BTV isolation was made in April (sheep) and 3 in June (1 each from sheep, cattle and goats); the remainder of the BTV isolations (a total of 359) were made in the months of July through early December. No isolations were made from January through March. Four serotypes of BTV (10, 11, 13 and 17) were isolated from all species tested (sheep, goats and cattle) and Culicoides variipennis. The serotypes isolated from C. variipennis correlated with the serotypes isolated from livestock in given areas. Multiple serotypes were isolated from single herds, flocks and individual animals on single given days. In addition to multiple serotypes being isolated, extensive heterogeneity in the electrophoretic mobility of the RNA genome segments was observed. These different migration patterns (electropherotypes) were observed between and within serotypes. No single serotype could be identified by a given pattern. No clinical disease was associated with BTV infection of cattle. Clinical disease was observed in infected sheep; however, BTV was also isolated from flocks with no overt clinical signs of disease. No reproductive problems could be associated with BTV in cattle in this endemic study area; however, BTV infection of pregnant sheep appeared to be associated with abortion and birth of dummy lambs in certain flocks. PMID- 2989908 TI - Molecular epidemiology of two US orbiviruses: bluetongue virus and epizootic hemorrhagic disease virus. AB - The degree of relatedness between 4 US serotypes (10, 11, 13 and 17) of bluetongue (BT) virus (BTV) were determined by both oligonucleotide fingerprint analyses of double-stranded (ds) RNA as well as tryptic peptide analyses of viral coded polypeptides. Similar studies were undertaken using different alternate isolates of particular serotypes as well as 2 serotypes (1 and 2) of epizootic hemorrhagic disease (EHD) virus (EHDV), a closely related serogroup of orbiviruses. The results indicate that all BT viruses are genetically related to each other and although EHDV-1 and EHDV-2 originated from a common gene pool, the 2 serogroups, EHDV and BTV, are not genetically interactive. PMID- 2989909 TI - Epidemiologic implications of the genetic variations of bluetongue virus. AB - The segmented RNA genome of bluetongue (BT) virus (BTV) provides for considerable diversity. This diversity has been seen in biochemical and biophysical analyses of the numerous strains of BTV as well as in clinical and immunologic responses of ruminant animals to BTV infection. This report describes the preliminary characterization of a unique BTV serotype 11 population recovered during 4 months in 1982 from 40 naturally infected animals including cattle, sheep and a goat. The strains of BTV serotype 11 were mild in their pathogenicity for the ruminants as no clinical signs of disease were seen. Infected cattle did not always develop detectable precipitating (P) or neutralizing (N) antibody during or after the infection. A better understanding of the epidemiology of BT may result from studies that include host and vector studies along with biochemical and biophysical characterization of the infecting BTV population. PMID- 2989910 TI - Research on the control of bluetongue in livestock by vector suppression. AB - From 1979-83, research has been conducted along the South Platte River drainage system northeast of Denver, CO, to develop and evaluate techniques of wide-area vector suppression for the control of bluetongue (BT) disease in ruminant livestock. Water management and chemical treatment at larval development sites have resulted in a decrease in the activity of Culicoides variipennis (Coquillett) in the 100 km2 treatment area, and there is some correlation between levels of vector activity (as reflected by light trap catches) and the levels of BT in livestock as measured by serologic and virologic evaluations. PMID- 2989911 TI - Vaccines for bluetongue and other orbiviruses from a regulatory viewpoint. PMID- 2989912 TI - Use of a quadrivalent modified-live bluetongue virus vaccine in wildlife species. AB - Three hundred and twenty-seven animals comprised of deer, mouflon sheep and bighorn sheep were vaccinated with an experimental quadrivalent, modified-live, bluetongue virus (BTV) vaccine. No untoward effects due to the vaccine were noted nor were abortions observed in vaccinated pregnant mouflon sheep. Precipitating antibodies to BTV and epizootic hemorrhagic disease (EHD) virus were identified in the serum of several animals prior to vaccination. Virus neutralizing antibodies to 3 of the 5 serotypes of BTV which exist in the US were identified in the serum of 2 peninsular bighorn sheep. PMID- 2989913 TI - Immune response of mice and sheep to bluetongue virus inactivated by gamma irradiation. AB - Gamma irradiation is being tested as a means of inactivating bluetongue virus (BTV) for use in vaccines. Exposure of BTV 17 to various levels of irradiation revealed that a dose of approximately 0.6 megarad was required to reduce the virus titer by one log10, or 90%. To test the immunogenicity of irradiated BTV, mouse brain passaged virus and concentrated cell culture passaged virus were inactivated by 6 megarads of gamma irradiation, and vaccines were prepared by emulsifying the virus preparations in equal volumes of a modified incomplete Freund's adjuvant. These vaccines stimulated the production of neutralizing antibodies in mice and sheep, a cell mediated immune response in mice, and a protective immune response in sheep. The results suggest that gamma irradiation would be an effective means of inactivating BTV for the preparation of vaccines. PMID- 2989915 TI - Control of bluetongue virus spread by embryo transfer. PMID- 2989914 TI - Potency and efficacy of inactivated bluetongue virus vaccines. AB - Bluetongue virus (BTV) was chemically inactivated and was shown to elicit neutralizing antibodies in vaccinated rabbits and sheep. In sheep, the neutralizing antibody was shown to be protective by immunity challenge with virulent virus. These studies have shown the feasibility of safe and effective inactivated vaccines for bluetongue (BT) disease of sheep. PMID- 2989916 TI - WHO/FAO Working Team Report: pathology. PMID- 2989918 TI - WHO/FAO Working Team Report: virology. PMID- 2989917 TI - WHO/FAO Working Team Report: entomology. AB - A summary of the more important concerns of the Working Team with particular reference to bluetongue (BT) virus (BTV), is as follows: With the exception of Australia, the US, and possibly parts of Africa, there are almost no concrete data that could be used to direct control efforts against the responsible vector(s) of BTV in a specific geographic area. In Australia, a broad plan of attack yielded data that showed that members of the subgenus Avaritia are the primary vector species. It has been clearly shown that Culicoides variipennis (Coq.) is the dominant vector species for most of the US. In the Middle East and certain areas of Africa, isolations from possible vector species indicated that C. imicola K., again probably a species complex and belonging to the subgenus Avaritia, is one of the likely vectors. Except for some subgenera, the taxonomy for much of the Culicoides remains in disarray. The subgenus Oecacta for example still remains a catchall for many species. Although a number of proposals have been made for delineating the species of the C. (Monoculicoides) variipennis species complex, none have been initiated because of the lack of funds. The Working Team strongly feels that the lack of definition of important species complexes is the result of short-term planning that does not consider the fact that efficiency can be increased and a great deal of monies saved by directing control toward the specific species or populations thereof that are responsible for a problem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989919 TI - "White eye calf" syndrome in Oregon associated with bluetongue and epizootic hemorrhagic disease viruses. PMID- 2989920 TI - WHO/FAO Working Team Report: molecular virology. PMID- 2989921 TI - WHO/FAO Working Team Report: immunology. PMID- 2989922 TI - WHO/FAO Working Team Report: epizootiology. PMID- 2989923 TI - The history of bluetongue. PMID- 2989924 TI - WHO/FAO Working Team Report: integrated disease management. PMID- 2989925 TI - Effect of bluetongue virus on reproduction in sheep and cattle. AB - Bluetongue virus (BTV) has been shown to be arbortigenic and teratogenic. After transmission of BTV under natural and experimental conditions the consequences of vertical transmission in pregnant sheep or cattle are variable. Factors that influence reproductive consequences are the stage of gestation, characteristics of the virus, source and concentration of virus inoculum, placentation, season and the method and route of infection. Reproductive consequences vary greatly in degree but in general include infertility, abortion, mummification of the fetus, stillbirths and congenital anomalies and dysfunctions in the live offspring. Immunological unresponsiveness, sporadic viremia and development of "late disease" were consequences of vertical transmission in offspring of infected cattle. Perpetuation of BTV through 3 generations in cattle was documented. The presence of BTV in semen of infected bulls has been demonstrated. PMID- 2989926 TI - WHO/FAO Working Team Report: international impact. PMID- 2989927 TI - Roundtable discussion on current policies regulating the international movement of animals and germplasm. PMID- 2989928 TI - Perspectives and recommendations for future research. PMID- 2989929 TI - Perspectives for future communications between research and regulatory agencies internationally. PMID- 2989930 TI - Orbivirus infection of the bovine fetus. AB - Direct inoculation of 85-125 day bovine fetuses with 4 serotypes of bluetongue virus (BTV) or 1 serotype of epizootic hemorrhagic disease virus (EHDV) resulted in either fetal death or teratogenesis. Three serotypes of BTV, and 1 of EHDV caused a similar cavitating encephalopathy, the severity of which likely reflected gestational age of the fetus at infection. Virus-specific antibody was readily detected in precolostral serum samples from all inoculated fetuses that survived until term, whereas, virus was not recovered from any of these calves. Calves with severe Orbivirus induced cerebral malformations would have poor viability, and they could not be expected to have any significance as virus reservoirs. PMID- 2989931 TI - Bluetongue virus and embryo transfer in cattle. AB - Embryos recovered nonsurgically from donor cattle infected with bluetongue virus (BTV) were transferred to seronegative, uninfected recipients. High titers of BTV were found in donor blood at the time of embryo recovery and BTV could be isolated from the recovery medium obtained from many of the donors. Bluetongue virus was not isolated from the blood of any of the 39 recipients that received embryos from infected donors, and none of these recipients seroconverted within 60 days of transfer. There was no evidence that BTV was transmitted to the early embryo in viremic donors. These results indicate that, under standard conditions of embryo transfer, transmission of BTV from infected donors to recipients is unlikely to occur. PMID- 2989932 TI - Seminal shedding of bluetongue virus in experimentally infected bulls. AB - Twenty bulls were inoculated with bluetongue virus (BTV) to study the frequency, duration and pathogenesis of seminal shedding of the virus. BTV was isolated from 19 of 181 ejaculates collected from these bulls during the period of viremia, but from none of 79 ejaculates collected after the viremia. Three categories of bulls were recognized on the basis of frequency and duration of seminal shedding of BTV. Virus was never isolated from semen in a majority (13 of 20) of bulls. In another group (5 of 20), virus was isolated from semen in low titer and from only 1 to 3 ejaculates. However, 2 bulls did shed relatively large quantities of virus in semen over a period of 2 to 3 weeks. Seminal shedding of BTV in infected bulls closely followed peak virus titers in blood and in no case was virus isolated from semen without its concurrent isolation from blood. Semen was processed and frozen from 1 bull that shed BTV. Six of 9 heifers inseminated at estrus with this semen became pregnant, and 3 of 9 became viremic and seroconverted. An additional heifer developed antibody to the virus, but did not become detectably viremic. No evidence of fetal infection was found in the heifer that became pregnant and infected, or in 5 heifers that became pregnant, but did not became infected. PMID- 2989933 TI - Unusual observations on a serologically negative bluetongue virus infected bull. AB - A Holstein bull named "Regency" born in New York in July 1968, maintained in an artificial insemination (AI) stud for 9 years, unremarkable from the point of view of health history or semen production, was discovered "by accident" to have bluetongue (BT) virus (BTV) in his semen. A microscopic study in 1975 and 1977 revealed unusual cytoplasmic vacuoles in the sperm of "Regency" prompting us to send semen to A. J. Luedke, USDA-ARS, Denver, Colorado, USA, to attempt virus isolation. Serotype 13 BTV was isolated from the semen. Serum sent with the semen was negative for antibody by the agar gel immunodiffusion (AGID) and complement fixation (CF) tests. Annual AGID serologic tests begun in 1975 and continuing to the present have been negative for antibody to BTV for all bulls in this AI stud. "Regency" was moved to Auburn, Alabama, USA in 1978 and further studies were performed. The bull was maintained with the Veterinary School herd. The BT serologic status of the cattle did not change during the year the bull was in the herd nor were the cows he bred or their calves serologically positive for BT. Studies on insemination of cattle with experimentally contaminated semen indicated that 5X10(4) infectious doses of BTV were required for infection. These results suggest that since "Regency's" semen did not infect cows, the semen probably contained less than this amount of virus. The rete testis was cannulated, rete fluid collected, a testis removed and virus isolation attempted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989935 TI - Interaction between prostaglandin E1 and forskolin in modulation of cyclic AMP levels in rat epididymal adipocytes. AB - The interaction between the diterpene, forskolin and prostaglandin E1 on cAMP levels in isolated adipocytes of the rat was examined. Various doses of Prostaglandin E1 in the presence of 1-methyl-3-isobutylxanthine (2 mM) blocked the increase in the intracellular level of cAMP elicited by forskolin (0.1 microM or 5 microM). Interestingly, in the absence of 1-methyl-3-isobutylxanthine (or an adenosine blocker), or in the presence of 1-methyl-3-isobutylxanthine (2 mM) and forskolin (30 microM), prostaglandin E1 (1 nM-100 microM) exhibited biphasic dose response kinetics. The stimulatory effect of PGE1 (greater than 1 microM) in presence of forskolin (30 microM) may be a consequence of prostaglandin receptor modification following endogeneous release of adenosine and prostaglandins by adipocytes stimulated to high levels of cAMP accumulation due to 30 microM forskolin. PMID- 2989934 TI - Multimolecular forms of oxytocinase activity in the human uterus and their inhibition by prostaglandins and cyclic GMP. AB - Oxytocinase (EC 3.4.11.3) activity was measured in uterine homogenates from pregnant and non-pregnant women using S-benzyl-L-cysteine-p-nitroanilide (BCN) and L-leucine-p-nitroanilide (LN) separately as substrates. There was no significant difference between the specific activity of the enzyme in the pregnant and non-pregnant uterus. The uterine homogenates were also subjected to gel filtration on Ultrogel AcA 22 column and two hydrolyzing activity peaks from the pregnant uterus (PUI, PUII) and three from the non-pregnant uterus (UI, UII, UIII) were obtained. Although the pattern of hydrolysis of the two substrates by the enzyme peaks was similar, the hydrolysis of LN was greater than that of BCN. PUII, UII and UIII showed higher affinity for the substrates than PUI and UI, and the affinity of all isoenzymes was significantly greater for LN than BCN. These uterine oxytocinases appeared to be metallo-aminopeptidases and were potently inhibited by Cu2+, Zn2+ and prostaglandins (PGs). Cyclic GMP (cGMP) and its 8 bromo derivative, however, inhibited only PUI and UI. The divalent cations and PGs were equipotent against the hydrolysis of both substrates, but cGMPs were more effective against BCN hydrolysis. All of these inhibitions depended on the pH of the incubation medium and types of the inhibition by PGs and cGMPs were mixed and uncompetitive respectively. The implications of these findings in relation to human parturition have been discussed. PMID- 2989936 TI - Infectious-inflammatory changes in cyclic AMP levels and in their regulation by prostaglandins in human peritoneal macrophages. AB - Peritoneal macrophages of renal patients on continuous ambulatory peritoneal dialysis (CAPD) have been collected when CAPD was without complications, during an intercurrent infectious peritoneal inflammation and after recovery. Levels of cyclic AMP, release of cyclo-oxygenase metabolites, and responsiveness, in terms of cyclic AMP elevation, to either PGE2 or to DC-PGI2 (a stable analogue of PGI2) were examined. Peritoneal inflammation was associated with a sharp drop in cyclic AMP, which was restored after recovery. Production of TXA2, PGI2 and PGE2 parallelled the direction of changes in cyclic AMP levels, except, that release of PGE2 entirely failed to recover. Macrophages during the uncomplicated stage of CAPD proved more responsive to DC-PGI2 than to PGE2. During inflammation the cells displayed a marked increase in sensitivity towards PG stimulation. Improved sensitivity was more pronounced with PGE2 than with DC-PGI2 and so the original difference between responsiveness of the cells to the PGs was abolished. Several findings are compatible with the view that endogenous PGI2 governs the cyclic AMP levels in human non-inflammatory peritoneal macrophages. However, during infectious-inflammation the cells undergo changes which render a reduced production of PGI2 insufficient to explain the drop in cyclic AMP. PMID- 2989937 TI - Prostacyclin infusion does not prevent ACTH-induced hypertension in sheep. AB - The present experiments examine the hemodynamic effects of an intravenous infusion of prostacyclin on the development of ACTH-induced hypertension in conscious sheep. Prostacyclin was infused at either 0.01 microgram/kg min-1 for 9 days or 0.25 microgram/kg min-1 for 4 days. At 0.01 microgram/kg min-1 prostacyclin had no effect on blood pressure in normotensive sheep or on the development of ACTH hypertension. Infusion at 0.25 microgram/kg min-1 increased heart rate, cardiac output and plasma renin concentration and decreased stroke volume and peripheral resistance in normotensive sheep. Despite these effects it did not prevent development of ACTH-induced hypertension. It is unlikely on the basis of these results that glucocorticoid-induced inhibition of vasodepressor prostacyclin and resulting increase in pressor responsiveness to circulating agonists is the primary cause of ACTH induced hypertension in sheep. PMID- 2989939 TI - Lithium effects on position learning with exploratory and aversive goal-box conditions. AB - Under conflict conditions in which reinforcement occurred within mildly aversive white goal-boxes, the chronic administration of lithium chloride (20 meg/l, in drinking water) to rats, while reducing avoidance of the goal boxes, impaired acquisition of an exploration-reinforced position discrimination. These results were interpreted as supporting the concept that lithium salts reduce the individual's reaction to general environmental stimulation--an effect which may account for its therapeutic action in the treatment of manic disorders. PMID- 2989938 TI - Increased sensitivity to serotonergic agonists after repeated electroconvulsive shock in rats. AB - The effect of single and repeated electroconvulsive shock (ECS) (once daily for 7 days) on head twitches produced by 5-HT agonists (LiCl, 5-hydroxytryptophan; 5 HTP and 5-methoxytryptamine; 5-MT) was investigated 1 hr, 24 hr, 5 days and 10 days after the last ECS, while locomotor activity induced by serotonergic agonists (fenfluramine, 3-chlorophenylpiperazine; m-CPP) and antagonists (metergoline, cyproheptadine) was only investigated after 24 hr. 5HT and 5-HIAA concentrations were measured 0.5, 1 and 24 hr after a single ECS and up to 10 days after repeated ECS. Head twitches induced by LiCl were significantly depressed 1 hr after both single and repeated ECS. The number of head twitches produced by LiCl, 5-HTP or 5-MT given 24 hr after single or repeated ECS did not change but it rose significantly 5 and 10 days after the last shock. Repeated ECS increased locomotor activity 24 hr after the last shock. This increase was significantly enhanced by serotonergic antagonists. Biochemical assays showed that a single ECS did not significantly change brain 5-HT and 5-HIAA concentrations 0.5, 1 or 24 hr after the ECS. On the other hand, repeated ECS raised brain 5-HIAA 0.5, 1 and 24 hr or 5 and 10 days and 5-HT 0.5 hr after the final ECS. It is concluded that a single or repeated ECS both depress the serotonergic system response to LiCl but repeated ECS facilitates the response to serotoninomimetics. PMID- 2989940 TI - Autoradiographic localization of kappa opiate receptors in CNS taste and feeding areas. AB - Recent evidence suggests that kappa opiate receptors may play a key role in the regulation of appetite. Such evidence implies that kappa receptors might be localized within specific brain areas known to regulate ingestive behaviors. On the basis of this implication we employed an in vitro film autoradiographic technique using 3H-ethylketocyclazozine as ligand to identify putative kappa receptors within CNS "taste" nuclei and surrounding areas. Coronal cryostat sections of rat brain were incubated with ligand in the presence of D-Ala2, D Leu5-enkephalin (DADLE) and morphine, apposed to LKB Ultrofilm for 60 days, processed and kappa receptor densities evaluated with the aid of a hand held photometer and video image analyzer. Highest kappa receptor densities were found within various gustatory and feeding sites including the rostral pole of the nucleus of the solitary tract, parabrachial nuclei, ventral posterior and medial portions of the thalamus, medial hypothalamus, medial nuclei of the amygdala and bed nucleus of the stria terminalis. Various other midline and medial limbic areas also showed significant kappa densities. PMID- 2989943 TI - Clonazepam-induced hyperphagia in nondeprived rats: tests of pharmacological specificity with Ro5-4864, Ro5-3663, Ro15-1788 and CGS 9896. AB - Nondeprived male rats were familiarised with daily 60 min access to a highly palatable diet, consisting of powdered rat diet, sweetened condensed milk and water. Clonazepam (0.625-5.0 mg/kg, IP) produced a substantial increase in food consumption within the first 30 min of access. The increase was similar across all dose conditions, suggesting that a maximal effect may have been achieved with a dose as small as 0.625 mg/kg. The hyperphagia induced by clonazepam was reversed by the benzodiazepine receptor antagonist, Ro15-1788 (5.0-20.0 mg/kg), indicating that the effect was benzodiazepine receptor-mediated. Treatments with the peripheral-type benzodiazepine agonist, Ro5-4864, did not induce a hyperphagic response. Instead, food consumption was significantly depressed following the administration of Ro5-4864 at 20 and 40 mg/kg, IP. A comparison of the clonazepam and Ro5-4864 data suggests that benzodiazepine-induced hyperphagia is mediated by central-type benzodiazepine binding sites. The pyrazoloquinoline, CGS 9896, binds with high affinity to benzodiazepine sites and has recently been described as a nonsedating anxiolytic. CGS 9896 (2.5-20.0 mg/kg, administered either IP or PO) did not affect consumption of the highly palatable diet. In consequence, anxiolytic and hyperphagic effects of drug actions at benzodiazepine receptors may be dissociated in the case of this compound. The atypical 1,4 benzodiazepine, Ro5-3663, a GABA antagonist which may act at the picrotoxinin site, produced a dose-related reduction in food consumption. Comparison with the results for Ro5-4864 rules out an interpretation for the anorexia in terms of anxiogenic effects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989941 TI - A possible role for beta-adrenergic receptors in the expression of audiogenic seizures. AB - DBA/2 mice are genetically prone to audiogenic seizures and, when compared with seizure resistant C57BL/6 mice, were found to have an increased density of beta adrenergic receptors in their midbrain at the age of peak seizure susceptibility. Propranolol, a beta-receptor blocking agent, attenuated all stages of the seizure syndrome. However, a comparison of the effects of its d- and l-isomers suggested that propranolol's anticonvulsant activity was due to its local anesthetic-like action. Pindolol, a more potent beta blocker that is at least 100 times less potent than propranolol with respect to local anesthetic-like activity, produced anticonvulsant effects in approximately the same systemic dose range as propranolol. This indicates that pindolol's anticonvulsant activity could be due to beta blockade and, taken together, these data suggest that beta-adrenergic receptors may play a role in the expression of audiogenic seizures in these animals. PMID- 2989942 TI - Effects of phencyclidine (PCP)-like drugs on turning behavior, 3H-dopamine uptake, and 3H-PCP binding. AB - In this study representatives from three chemical classes which are known to produce a phencyclidine (PCP)-like discriminative stimulus cue in rats were tested for their ability to inhibit synaptosomal uptake of 3H-dopamine (3H-DA) and to compete for a binding site labeled with 3H-PCP. Although there was a good correlation between these two in vitro activities within the arylcycloalkylamine class (PCP, N-ethyl-phenylcyclohexylamine (PCE), and ketamine) it did not hold when representatives from the benzomorphans, N-allynormetazocine (NANM), cyclazocine (CYCL), and ethylketocyclazocine (EKC), or a substituted dioxolane (etoxadrol) were included. At some dose each of these drugs with the exception of EKC also produced ipsilateral turning in rats with a unilateral 6-hydroxydopamine lesion of the substantia nigra. This effect was also not well correlated with inhibition of 3H-DA uptake. However, a significant correlation was found to exist between turning behavior and affinity for the putative PCP/sigma receptor. The possibility that a non-dopaminergic mechanism involving the PCP/sigma receptor underlies the ability of these agents to induce turning behavior is discussed. PMID- 2989944 TI - Pimozide decreases the positive reinforcing effect of sham fed sucrose in the rat. AB - Rats with chronic gastric cannulas were intraperitoneally injected with the dopamine receptor antagonist pimozide (0.25 mg/kg) before sham feeding 5, 10, 20, or 40% (w/v) sucrose solutions. Amount of sham intake after control injections increased as a function of sucrose concentration. At each concentration, the rate of sham feeding was greatest during the initial 3 min of sham feeding and subsequently decelerated. Pimozide inhibited sham feeding rate, and the temporal pattern of decreases in sham feeding rate after pimozide were similar to those produced by decreasing the concentration of sucrose sham fed. These data extend previous reports of the inhibitory effect of pimozide on ingestion of sweet fluids by eliminating the possibility that the effect was the result of pimozide facilitating postingestional inhibitory mechanisms. Further, pimozide did not appear to produce fatigue or sedation, or to reduce the rats' motor capacity to sham feed. Therefore, these data are consistent with the hypothesis that central dopaminergic synaptic activity mediates the reinforcing effects of sweet taste that drive sham feeding. PMID- 2989945 TI - Continuous infusion of naloxone: effects on behavior and oxygen consumption. AB - Twenty-eight hours of endorphin receptor blockade by subcutaneous naloxone infusion produced behavioral and respiratory symptoms resembling opiate abstinence syndrome. Rats were implanted subcutaneously with two Alzet osmotic minipumps delivering 0.7 mg/kg per hour naloxone or with two control minipumps containing distilled water only. They were observed for 10 minutes under blind conditions at 16 and 28 hours post-implantation. The naloxone-infused rats showed significantly more wet dog shakes, abdominal writhes and overall abstinence-like symptoms than did the control rats. These symptoms decreased after 28 hours despite continued naloxone infusion. Acute administration of naloxone failed to produce abstinence-like symptoms, even when combined with the trauma of carrying two implanted water-filled minipumps for 28 hours. In another experiment, naloxone-infused rats showed a highly significant 53.4% elevation of O2 consumption over water-infused control rats in a pure O2 atmosphere at 28 hours after implantation. This difference disappeared at 48 hours post-implantation. In contrast to the effect of naloxone infusion, acute administration of three different doses of naloxone failed to significantly increase O2 consumption. PMID- 2989946 TI - Methylxanthine discrimination in the rat: possible benzodiazepine and adenosine mechanisms. AB - Rats were trained to discriminate either caffeine or theophylline from saline using a two-lever discrimination paradigm. Since methylxanthines have been found to interfere with agonist binding at both adenosine and benzodiazepine (BDZ) receptors, chlordiazepoxide (CDP) and L-PIA (an adenosine analog) were tested for generalization to and blockade of both xanthine cues. Neither L-PIA nor CDP generalized to either xanthine cue, although both produced dose-related decreases in response rate. CDP, but not L-PIA, produced dose-related decreases in drug lever responses when combined with training doses of caffeine or theophylline. Response rates indicated a complex interaction between the xanthines and both L PIA and CDP. When combined with the caffeine training dose, pentobarbital also produced a dose-dependent decrease in response rate but not in drug lever choices. Finally, papaverine generalized to the caffeine cue in a dose-dependent fashion. In a second experiment, rats trained to discriminate CDP from saline showed no generalization in L-PIA tests. CDP-appropriate responding was not significantly affected when the CDP training dose was combined with caffeine. These data indicate that: (a) methylxanthine interactions with L-PIA and CDP on response rate likely involve blockade of adenosine mechanisms; (b) the xanthine cue does not appear to depend on interactions with adenosine receptors; and (c) the xanthine cue may involve effects on cyclic AMP activity and/or interaction with the BDZ/GABA receptor complex. PMID- 2989947 TI - Distribution and function of peripheral alpha-adrenoceptors in the cardiovascular system. AB - alpha-Adrenoceptors may be subdivided based on their anatomical distribution within the synapse. Presynaptic alpha-adrenoceptors are generally of the alpha 2 subtype and modulate neurotransmitter liberation via a negative feedback mechanism. Postsynaptic alpha-adrenoceptors are usually of the alpha 1-subtype and mediate the response of the effector organ. Although this "anatomical" subclassification is generally applicable, many exceptions exist. A more useful classification of alpha-adrenoceptor subtypes is based on a pharmacological characterization in which selective agonists and antagonists are used. Peripheral alpha-adrenoceptors are critical in the regulation of the cardiovascular system. Postsynaptic alpha-adrenoceptors in arteries and veins represent a mixed population of alpha 1/alpha 2-adrenoceptors, with both subtypes mediating vasoconstriction. In the peripheral arterial circulation, postsynaptic vascular alpha 1-adrenoceptors are found in the adrenergic neuroeffector junction, whereas postsynaptic vascular alpha 2-adrenoceptors are located extrajunctionally. In the venous circulation, it appears that alpha 2-adrenoceptors may be predominantly junctional, whereas alpha 1-adrenoceptors may be predominantly extrajunctional. It has been proposed that junctional alpha-adrenoceptors will respond predominantly to norepinephrine liberated from sympathetic neurons, whereas extrajunctional alpha-adrenoceptors likely respond to circulating catecholamines. The functional role of extrajunctional alpha-adrenoceptors may be more important in disease states such as hypertension and congestive heart failure where circulating levels of catecholamines may be high and contribute to the maintenance of elevated vascular resistance. alpha 2-Adrenoceptors are also associated with the intima and may play a role in the release of an endogenous relaxing factor from the endothelium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2989948 TI - Heterogeneity of alpha-2 adrenergic receptors. AB - Alpha adrenergic receptors are subdivided into alpha-1 and alpha-2 subtypes on the basis of their pharmacologic properties. An evaluation of data from radioligand binding and functional experiments indicates that the receptors classified as alpha-2 are not a homogeneous group. The best example of this heterogeneity is the differences in the pharmacologic properties of alpha-2 receptors in rodent and non-rodent mammalian species. Prazosin generally has a high affinity for rodent alpha-2 receptors, but a low affinity for non-rodent alpha-2 receptors, while oxymetazoline is more potent at non-rodent alpha-2 receptors. A definition of alpha-2 adrenergic receptor subtypes is proposed with prazosin having a lower affinity (200-300 nM) at alpha-2A receptors and a higher affinity (5-10 nM) at alpha-2B receptors. Using this definition, the human platelet appears to have only the alpha-2A subtype, while all the receptors in the neonatal rat lung are of the alpha-2B subtype. The rat brain has roughly equal amounts of alpha-2A and -2B receptors while in the rat submandibular gland, about 85% of the receptors are alpha-2A. The ability to pharmacologically define putative alpha-2 adrenergic receptor subtypes should promote the development of additional subtype selective drugs which will increase our understanding of adrenergic pharmacology and may provide new therapeutic approaches. PMID- 2989949 TI - The alpha-adrenergic receptor: radiohistochemical analysis of functional characteristics and biochemical differences. AB - The partial agonist [3H]para-aminoclonidine was used to label alpha 2-adrenergic binding sites in intact sections of the rat central nervous system using in vitro labeling receptor autoradiographic techniques. The distribution of alpha 2 agonist binding sites closely parallels the reported distribution of noradrenergic and adrenergic cell groups and their terminal fields, particularly the projections of the medullary catecholamine neurons. This distribution of alpha 2 binding sites confirms physiological studies which indicate that the anti hypertensive actions of alpha 2-agonist compounds are mediated centrally in medullary and spinal centers involved in the control of parasympathetic and sympathetic outflow. Further, the high concentrations of alpha 2 binding sites in pontine and limbic areas such as the locus coeruleus, parabrachial nucleus, dorsal raphe, hypothalamus, amygdala, bed nucleus of the stria terminalis, septum and entorhinal cortex offer an anatomical basis for understanding the anxiolytic and antidepressant actions of drugs like clonidine. The antagonists [3H]prazosin and [3H]WB4101 were used to study the distribution of alpha 1-adrenergic binding sites in the rat forebrain and biochemical studies were performed to analyze the marked differences that were initially seen in the distribution of [3H]prazosin and [3H]WB4101 binding sites. Several pieces of evidence derived from both biochemical and autoradiographic studies suggest that [3H]prazosin and [3H]WB4101 act at distinctly different binding sites. However, both sites may represent components of an alpha 1-adrenergic receptor-effector complex since a high degree of overlap was seen in the binding site distribution of these two ligands and since kinetic interactions could be demonstrated in at least one region of the brain, the hippocampus. Differences noted in the relative displacements of [3H]prazosin and [3H]WB4101 binding in various forebrain regions could reflect differences in the coupling efficiency of the [3H]prazosin and [3H]WB4101 component of the hypothesized complex. Further, in some regions, [3H]WB4101 labeled a binding site that is different from the alpha 1-receptor. Thus, [3H]prazosin and [3H]WB4101 binding sites seen in forebrain regions such as lamina V of the cortex, thalamic nuclei and dorsal raphe probably represent alpha 1-adrenergic receptors and confirm electrophysiological and biochemical studies which demonstrate that adrenergic transmission in these regions can be mediated through an alpha 1-receptor. PMID- 2989950 TI - Effects of quipazine and methysergide on play in juvenile rats. AB - Social play of juvenile rats was analyzed following administration of either the serotonin receptor agonist quipazine (1, 2.5, 5 and 10 mg/kg) or the antagonist methysergide at the same doses. Quipazine reduced pinning at all doses, while methysergide did so at only the highest two. An interaction study using the lowest doses of the agents used above, indicated that methysergide could reverse quipazine induced reductions of play. Thus, the quipazine effect was probably mediated through a serotonin receptor; however, the role of serotonin in play appears to be a general modulatory one rather than a specific influence on play. PMID- 2989951 TI - Progesterone in rat brain: modulation of beta-adrenergic receptor activity. AB - Cytosolic proteins binding specifically the progesterone analogue 3H R5020 can be detected in various areas of the central nervous system (CNS) of rat. Our study demonstrates that the concentration of progesterone receptors in the brain of adult, ovariectomized, female rats is maximal in frontal cortex and midbrain and lowest in the cerebellum. Short term administration of the hormone does not alter the beta-adrenergic receptor system, while a prolonged administration of progesterone results in an up-regulation of 3H-DHA binding in the frontal cortex. Such increase in binding activity is due to an increased number of beta adrenergic receptors. Furthermore, high doses of progesterone can also increase the number of beta-adrenergic binding sites in an "in vitro" system where brain slices are incubated for few hours in a physiological medium. PMID- 2989952 TI - Effect of captopril on Na-K ATPase and Mg ATPase activities of erythrocyte ghost membranes. AB - Captopril has a known hypotensive action derived from the inhibition of the converting enzyme (Kininase II). To investigate whether other mechanism may contribute to vasodilation, Na-K ATPase and Mg ATPase activities on ghost red blood cells membranes were examined in the presence of the drug. The results show a stimulation of Mg ATPase activity and an inhibition of Na-K ATPase activity in a concentration dependent manner. The latter action does not seem to be related to a vasodilator effect. PMID- 2989953 TI - Correlation between anti-hypertensive activity in rats and plasma angiotensin I converting enzyme (ACE) inhibition in mice following oral administration of ACE inhibitors. AB - Thirty-four ACE inhibitors were evaluated to determine the concentration giving 50% inhibition of purified rabbit lung ACE (IC50 microM) using benzyloxycarboxyl p-NO2-Phe-His-Leu as substrate, to determine the oral dose causing a lowering in blood pressure of 30 mm Hg (ED30 mumol/kg) in acute aortic coarctate (AAC) rats, and to determine inhibition of plasma ACE (PACE) activity in mice after oral dosing. Mouse PACE activity was determined with 14C-Hip-His-Leu as substrate one hour after oral dosing of 3 animals/group with 5 or 50 mumol ACE inhibitor per kg. Data from mice were expressed as percent of control group PACE activity. Least squares regression analysis showed the IC50 data to be poorly correlated with either rat data or mouse PACE data at 50 mumol/kg p.o. However, correlation was significant between log rat ED30 and mouse PACE at 5 (p less than 0.001, r = 0.570) and 50 (p less than 0.025, r = 0.387) mumol/kg p.o. Thus, the simple mouse plasma ACE determination after a dose of 5 mumol/kg is a convenient supplement to the AAC rat model for showing oral activity of ACE inhibitors. PMID- 2989954 TI - Characterization of the contractile activity of dopamine on the rat isolated seminal vesicle. AB - The mechanism of the contractile effect of dopamine (DA) on the rat isolated seminal vesicle was studied. Cocaine (10 microM/1 in the organ bath, 30 min before DA) and 6-OHDA (50 mg/kg i.v. 24 hr before removal of the seminal vesicle) almost completely prevented the contractile effect of DA. Drugs known to have an affinity for DA receptors or for alpha-adrenoceptors antagonized the contractile effect of DA, the rank order of potency being: prazosin greater than phentolamine greater than yohimbine greater than clonidine greater than sulpiride greater than apomorphine greater than haloperidol. The antagonism was in each case greater against DA than against noradrenaline (NA), used for comparison; selectivity for DA being highest in the case of prazosin and sulpiride. Taken together, these findings indicate that DA makes the rat seminal vesicle contract mostly by means of an indirect mechanism, binding presynaptic DA-receptors and, in part, presynaptic alpha-adrenoceptors as well; or, alternatively, binding presynaptic DA-receptors which have some links with alpha 2-adrenoceptors; the consequence being in either case the release of NA from sympathetic nerve endings. PMID- 2989955 TI - Effects of some intravenous anaesthetics on the contractile responses produced in the chick biventer cervicis skeletal muscle. AB - The effects of some intravenous anaesthetics, in low and high concentrations, on the contractile responses produced in the chick biventer cervicis skeletal muscle were studied using electrophysiological and pharmacological techniques. The anaesthetics used were methohexitone sodium, thiopentone, ICI 35 868 (propofol, diprivan), althesin and etomidate. The results showed that all these anaesthetics had a common mode of action at the chick neuromuscular junction. In low concentrations, they increased the amplitude of the indirectly-elicited twitch and tetanic contractions, whereas in high concentrations they reduced the evoked contractions and the contractures produced by depolarizing agents such as acetylcholine or tetraethylammonium. The results indicated that, in low concentrations, anaesthetics may stimulate skeletal muscle whereas in high concentrations they reduce the sensitivity of the postjunctional membrane to depolarizing drugs. The possibility that anaesthetics may block endplate ion channels was also discussed. PMID- 2989956 TI - [Molecular biology methods for the study of the cytochrome P-450 system and its potential applications]. PMID- 2989957 TI - Comparison between the luminol-dependent chemiluminescence of polymorphonuclear leukocytes and of the myeloperoxidase-HOOH system: influence of pH, cations and protein. PMID- 2989958 TI - Concept of natriuretic hormone. PMID- 2989960 TI - Target biting associated with a gustatory avoidance response. AB - Gustatory avoidance studies normally focus on the diminished occurrence of a consummatory behavior as the primary dependent variable. Several reports have described behaviors which accompany this gustatory avoidance response but no attempt has been made to automate collection of such data. In the following studies lithium chloride (LiCl) was administered to rats following saccharin consumption in a standard gustatory avoidance paradigm. The rats also had the opportunity to make noncontingent target biting responses on an inanimate target. It was observed that there was a significant decrease in saccharin intake following its pairing with LiCl. High target biting rates were associated with this avoidance response and, to a lesser degree, with initial target exposure. These observations are discussed in terms of paradigm contingencies. PMID- 2989959 TI - Dissociation of epinephrine's hyperglycemic and anorectic effect. AB - The contributions of alpha- and beta-adrenergic receptor mechanisms to epinephrine-induced hyperglycemia, to prandial hepatic glycogenolysis, and to epinephrine-induced inhibition of feeding were investigated in rats, because epinephrine's hyperglycemic and anorectic effect might be related to each other. Intraperitoneal injection of the selective alpha-adrenergic receptor antagonist phentolamine (0.5 mg/kg body weight) but not of the selective beta-adrenergic receptor antagonist propranolol (0.5 mg/kg body weight) reduced the hyperglycemic effect of a subsequent epinephrine injection (0.05 mg/kg body weight). In addition, injection of phentolamine (0.5 mg/kg b.w.) or injection of phentolamine plus propranolol (0.5 mg/kg b.w. each) similarly inhibited epinephrine-induced hepatic glycogenolysis. In contrast, phentolamine injection did not affect prandial hepatic glycogenolysis. Neither phentolamine nor propranolol injection was sufficient to affect the inhibition of feeding induced by subsequent epinephrine injection, but the combination of both receptor blockers completely abolished epinephrine's effect on food intake. These findings are not consistent with the idea that epinephrine-induced hypophagia is related to epinephrine's effect on hepatic carbohydrate metabolism. In addition, the data suggest that catecholamines are not the main contributors to prandial hepatic glycogenolysis in rats. PMID- 2989962 TI - Breast reconstruction following lumpectomy and irradiation. AB - The traditional approach to breast cancer has been the surgical mastectomy. The cosmetic deformity resulting from this procedure has led to the development of our current methods of breast reconstruction. The inherent attraction of lumpectomy and radiotherapy has been its equation with improved cosmesis. This procedure may be an effective alternative to surgery from the cancer perspective and does produce, in the majority of patients, an acceptable cosmetic result. However, this paper demonstrates that it may produce a result that is less optimal than mastectomy with postoperative breast reconstruction. The etiology of the increased deformity can either be the localized lumpectomy with marked internal derangement of breast volume or the radiation therapy itself. In addition, those who develop a combination require a solution to both. We describe three patients who illustrate these problems and believe that a greater awareness of these potential complications should be available to aid patients in choosing their cancer treatment. PMID- 2989961 TI - Intraventricular alloxan impairs feeding to both central and systemic glucoprivation. AB - Fourth ventricular alloxan injections impair the feeding response to systemically induced glucoprivation. However, the effect of alloxan on responses to centrally administered glucoprivic agents has not been assessed. Therefore, rats implanted with fourth ventricular cannulas were tested for feeding and glycemic responses to centrally administered 5-thioglucose (5TG, 120 micrograms/5 microliter) and subcutaneously administered 2-deoxy-D-glucose (2DG, 250 mg/kg, IP). Subsequently, alloxan (200 micrograms in 5 microliter), alloxan plus 3 M D-glucose, or the saline vehicle solution was injected into the fourth ventricle and the animals were then retested with systemic 2DG and intraventricular 5TG. Feeding responses to both centrally and systemically induced glucoprivic challenge were greatly impaired by alloxan treatment. Thus, feeding elicited by both centrally and systemically induced glucoprivation appears to be mediated by the same central neural substrate. The possibility that the alloxan-damaged cells are glucoreceptors is discussed. In contrast to glucoprivic feeding, the hyperglycemic response to glucoprivation was not impaired by alloxan. Therefore, the neural elements controlling this response may differ biochemically from those controlling feeding. PMID- 2989963 TI - Noradrenergic function and the cortisol response to dexamethasone in depression. AB - Abnormalities of the hypothalamic-pituitary-adrenal (HPA) axis and the noradrenergic system have been reported in depression. To study possible interrelations between these two systems, plasma free 3-methoxy-4 hydroxyphenylglycol (MHPG) was compared with the cortisol response to dexamethasone in 64 depressed patients. Postdexamethasone cortisol nonsuppressors had higher baseline plasma free MHPG values than did cortisol suppressors. Increased severity of some depressive symptoms was associated with increased post dexamethasone cortisol levels. These results indicate that depressed patients with dexamethasone nonsuppression have increased noradrenergic turnover. PMID- 2989964 TI - Genetic analysis of the effects of morphine on plasma cyclic nucleotides and locomotor activity in male mice. AB - A biometric analysis of morphine-induced changes in locomotor activity and plasma cyclic nucleotide levels was conducted in two inbred strains of mice, DBA/2N and C57BL/6N, and their F1 hybrid and backcross progenies. C57BL/6N was partially dominant over DBA/2N for morphine-induced motor activity, whereas DBA/2N was partially dominant over C57BL/6N with respect to the effect of morphine on the cyclic nucleotides. The strain difference in the effect of morphine on motor activity and on plasma cyclic GMP was shown to be genetically determined, whereas the genetic influence on plasma cyclic AMP was statistically not confirmed. The number of gene pairs which regulate the effect of morphine on motor activity was assumed to be more than one; however, the contribution of more than one segregating unit in the effect of morphine on plasma cyclic GMP was not demonstrated. PMID- 2989965 TI - Generalization of the discriminative stimulus properties of delta 9-THC to delta 9(11)-THC in rats. AB - Rats were trained in a water maze to discriminate between IP injections of 3 mg/kg delta 9-tetrahydrocannabinol (delta 9-THC) and its vehicle. Both delta 8- and delta 9(11)-THC were generalized to the training drug. In contrast to our observations in rhesus monkeys, where delta 9(11)-THC is at least 100 times less potent than delta 9-THC, delta 9(11)-THC was found to be only seven times less potent in the rat. Relative potencies, expressed as the dosage at which 50% of the animals gave drug responses (ED50) were 1.8 mg/kg and 12.2 mg/kg for delta 9- and delta 9(11)-THC respectively. Twenty-four hours after receiving 7 X ED50 = 12 mg/kg delta 9-THC the tests showed intermediate results when conducted with the training dosage; 4 X ED50 = 50 mg/kg delta 9(11)-THC 48 h prior to the training dosage of 3 mg/kg delta 9-THC completely blocked drug-appropriate responses. Coinjection of ED50 dosages of delta 9- and delta 9(11)-THC led to 90% drug responses, demonstrating the additivity of the cannabis-like effect of both cannabinoids. Differences in the individual sensitivity of the rats to the tested cannabinoids were observed. Findings are interpreted in terms of the receptor mechanism for cannabis-like activity. PMID- 2989968 TI - Assessment of the optical contributions to the age-related deterioration in vision. AB - Contrast sensitivities were measured in human observers of different ages in response to sinusoidal grating patterns generated by laser interferometer and cathode ray tube (c.r.t.). The former method bypasses the effects of the optical media to assess directly the contrast sensitivity of the retina/visual system, while the over-all assessment including optical media is made to the c.r.t. display. The ratio of the two sets of values gives the contrast ratio of the optical media. The latter assessment does not include the effects of wide-angle light scattering which were measured by comparing the responses of eyes with a Perspex lens implant or without any lens at all (i.e. without the major source of scattering) with natural eyes. With increasing age, c.r.t. contrast sensitivities remained steady until the sixth decade when they declined while laser interference fringe contrast sensitivities declined continuously, apart from some abnormally low results for teenage observers. In contrast, the optical contrast did not vary systematically with age nor did its rate of change with spatial frequency vary. The latter was always less than the rate for laser interference fringe contrast sensitivities indicating that the retina/visual system always sets the limit to visual resolution. Wide-angle light scattering was not found to contribute significantly to the changes in ageing nor did reduced retinal illumination, as reproduced by reduced pupil diameter or viewing through a neutral density filter. We therefore conclude that the age-related deterioration is primarily caused by changes within the central nervous system rather than the optical media, the transmission quality of which remains unaffected. PMID- 2989967 TI - Increase in plasma ACTH after dopaminergic stimulation in rats. AB - The effects of a dopaminergic agonist, apomorphine, and a dopaminergic antagonist, haloperidol, on plasma ACTH, and corticosterone levels were evaluated in adult male rats. Subcutaneous administration of apomorphine in the dose range of 50-500 micrograms X kg-1 significantly increased plasma corticosterone levels. Acute treatment with apomorphine (250 micrograms X kg-1) resulted in an elevation of plasma ACTH concentration, peak values being reached 15 min after the injection. The apomorphine-induced rise in plasma ACTH levels was completely inhibited by pretreatment with haloperidol (1 mg X kg-1). A stimulatory role for dopamine receptors in the control of pituitary ACTH release in the rat is suggested. PMID- 2989966 TI - Behavioral analysis of the effect of neurotensin injected into the ventral mesencephalon on investigatory and spontaneous motor behavior in the rat. AB - The present experiments examined in detail the behavioral response to microinfusions of neurotensin (NT) into the ventral tegmental area (VTA), substantia nigra (SN) and hippocampus (HPC). The behavioral apparatus consisted of an eight-hole box in which investigatory and spontaneous motor behavior were recorded. Three doses (0.175, 0.5, 4.0 micrograms) of NT were injected into the VTA. The main effect of NT was a strong augmentation of rearing (frequency and duration) both in the periphery and center of the arena, accompanied by a small increase in locomotion and decreased grooming. NT had no effect on the strategy, organization, or duration of exploration but did augment frequency of hole visits towards the end of the session. NT injected into the SN and HPC had no effect on investigatory and spontaneous behavior with the exception of an increase in peripheral locomotion after HPC-NT injections. The results are discussed in terms of a modulatory role of endogenous NT on mesolimbic dopamine neurons. PMID- 2989969 TI - Drug effects on the hydraulic conductivity of the isolated rabbit ciliary epithelium. AB - Three aspects of hydrostatically driven volume flow across the isolated rabbit ciliary epithelium have been examined. (1) Hydraulic conductivity was increased in a dose-dependent manner by amiloride, but not by furosemide, indicating that amiloride may interfere with junctional permeability. The absence of a furosemide effect suggests that regulation of passive fluid movement may be independent of net ionic fluxes of either chloride or bicarbonate. Ethacrynic acid had no effect on hydraulic conductivity at concentrations below 10(-2) M. At 10(-2) M, hydraulic conductivity was decreased, suggesting that fluid pathways may be linked to ionic pathways. (2) Successive in vivo daily treatments with adrenergic agonists result in changes in the intraocular pressure response. Dose-effect curves to in vitro agonists from eyes pre-treated on successive days indicate that isoprenaline caused a lack of response in hydraulic conductivity after 3 d of in vivo treatment, adrenaline caused responses after 2 d which were about half of those found on day 0, while phenylephrine and noradrenaline had no influence on hydraulic conductivity. These data suggest that beta-agonists cause a deactivation or desensitization of some moiety which is coupled with regulation of hydraulic conductivity in a manner similar to that seen for the beta-receptor coupled adenylate cyclase. (3) While cyclic GMP was without effect, various modifiers of cyclic GMP activity increased hydraulic conductivity. Alteration of cellular cyclic GMP levels appears to exert some regulation over ciliary epithelial permeability. PMID- 2989970 TI - Familial elevation of serum angiotensin converting enzyme. AB - We report here a familial clustering of elevated serum angiotensin converting enzyme (ACE) levels. The patient is a 58-year-old Japanese female who had been in excellent health until age 45 when she developed an occlusion of the left central retinal vein. She was otherwise in excellent health, and no laboratory abnormality except a marked elevation of serum ACE level (625 nmol/min/ml; normal range; 22-40 nmol/min/ml of serum) was found. Her blood pressure was within normal limits (140/80 mmHg). There was no evidence for the diagnosis of sarcoidosis, Gaucher's disease, leprosy, hyperthyroidism, diabetic retinopathy, or liver disease. One of her two sisters also showed a marked increase in serum ACE activity (303 nmol/min/ml), and remarkably high levels of serum ACE (276 and 294 nmol/min/ml) were demonstrated in both sons of this sister. All the members of this family have been in excellent health. The serum ACE activity was activated by chloride and cobalt ions, and inhibited by EDTA, captopril and rabbit antiserum to purified human plasma ACE. Thus our study showed a familial clustering of elevated serum ACE in individuals who did not have conventional disease patterns associated with elevated serum ACE. PMID- 2989971 TI - An unusual case of Wilson's disease. AB - A man of 61 with a 26-year history of progressive cerebellar ataxia was admitted to hospital. He was found to have chronic liver disease and died 22 days after admission. A diagnosis of hepatolenticular degeneration (Wilson's disease) was supported by clinical investigations and confirmed at autopsy, when tissue copper studies were performed. Several unusual features were present, including a unilateral Kayser-Fleischer ring, a hepatocellular carcinoma, peripheral neuropathy, pontine demyelination and calcification of neurones in the medulla. The significance of these findings is discussed with a review of the relevant literature. PMID- 2989972 TI - Bile duct necrosis: complication of transcatheter hepatic arterial embolization. AB - Bile duct necrosis because of transcatheter hepatic arterial embolization (THAE) in two patients with hepatocellular carcinoma is reported. Preoperative THAE was performed on 29 patients, and bile duct necrosis was experienced by two of the 29 (7%). In these two patients, gelatin (Gelfoam) powder was used as the embolus. Among the 24 whose embolus was clear, four were embolized with gelatin powder. Therefore, incidence of bile duct necrosis after THAE with gelatin powder was 50%. Because of the hazards of severe complications such as bile duct necrosis, we conclude that gelatin powder should not be used except for the THAE of no more than one segment of the liver. PMID- 2989973 TI - Detection of hepatic metastases by proton spectroscopic imaging. Work in progress. AB - Fourteen patients with hepatic metastases underwent magnetic resonance (MR) imaging using both the conventional spin-echo (SE) technique and the opposed phase of the proton spectroscopic imaging method. The opposed image showed more lesions than the conventional SE image in five patients and provided better contrast between the liver parenchyma and metastases in two patients. Four of these seven patients had associated fatty infiltration of the liver. When compared with the computed tomography (CT) scan, the opposed image either showed more lesions or provided better contrast in six patients, four of whom had fatty infiltration. More significantly, the MR image showed several 1-cm lesions not shown by the CT scan in one patient. Our study discloses the possible explanations for the increased sensitivity of the opposed image in detecting hepatic metastases. PMID- 2989974 TI - Intraoperative sonography: clinical usefulness in liver surgery. AB - In the past 4 years intraoperative sonography was performed on 83 patients with primary hepatic carcinoma, 11 with benign hepatic tumor, nine with intrahepatic lithiasis, five with metastatic hepatic carcinoma, and four with other benign hepatic diseases, for a total of 112 patients. Ultrasonography detected primary carcinoma in 80 of 83 patients (96.4%) and intrahepatic metastases in 30 of 33 patients (90.9%), as confirmed later in surgical specimens. Tumor thrombi in the portal vein were detected in nine of 13 patients (69.2%). In patients with intrahepatic lithiasis and benign space-occupying lesions, residual stones could be assessed easily and the nature and location of the lesions identified. Intraoperative sonography demonstrates intrahepatic ductal structures clearly and is the final diagnostic imaging procedure before surgery. PMID- 2989975 TI - Scrotal calculi: sonographic detection. AB - Scrotal calculi are freely moving concretions that lie in the space between the tunicae lining the scrotum and the testicles. They are believed to originate either as fibrinous deposits in the tunica vaginalis testis or as remnants of the appendix testis or appendix epididymidis that have undergone torsion and become freely movable. A sector real-time scanner with a 7.5-MHz transducer and a small parts scanner with a 10-MHz transducer were used to visualize scrotal calculi in two patients. The ultrasonic appearance is described along with the surgical findings, and the pathogenesis of this benign process is discussed. PMID- 2989976 TI - Selective hepatic angiography using a balloon catheter guide. AB - A 3-F balloon catheter guide was successfully used instead of a conventional guide wire for selective hepatic angiography in 30 patients. The technique provides an easy way to advance the catheter without complications. PMID- 2989977 TI - Effect of the thromboxane receptor antagonist EP 092 on endotoxin shock in the sheep. AB - The thromboxane receptor antagonist EP 092 inhibits the acute pulmonary vascular response to E. coli endotoxin in the anaesthetized, closed-chest sheep. The increase in the TXB2 level in arterial blood was not suppressed by EP 092. Intravenous infusion of the thromboxane mimetic 11,9-epoxymethano PGH2, but not PGF2 alpha, raises pulmonary artery pressure and lowers arterial pO2 similar to the endotoxin. Isolated strips of lobar pulmonary veins but not lobar arteries are contracted by low concentrations of 11,9-epoxymethano PGH2 - the effects are potently inhibited by EP 092. PMID- 2989979 TI - ACTH-releasing activity of urotensin I and ovine CRF: interactions with arginine vasotocin, isotocin and arginine vasopressin. AB - The release of ACTH from superfused dispersed goldfish anterior pituitary cells was examined to determine if the neurohypophyseal peptides arginine vasotocin (AVT), isotocin (IST) or arginine vasopressin (AVP) potentiate the ACTH-releasing activities of the structurally homologous peptides urotensin I (UI) or ovine corticotropin-releasing factor (CRF). The ACTH-releasing activities of the neurohypophyseal peptides and UI or CRF were additive. AVT, IST or AVP failed to potentiate the ACTH-releasing activity of UI or CRF. These results suggest that in teleost fishes neurohypophyseal peptides have intrinsic ACTH-releasing activity but, unlike mammals, do not potentiate the release of ACTH evoked by CRF, or by the piscian CRF-like peptide, UI. PMID- 2989978 TI - Leukocytic functions in burn-injured patients. AB - Anergy associated with an increase in suppressor helper T cell (Tc) ratio and a decrease in natural killer (NK) is one main cause of death following thermal injury (Tl). Recently, in vitro studies have shown that LTB4 can induce human Tc to exert suppressor cell activity, and incubation of lymphocytes with LTB4 for 24 hours significantly suppressed NK cell activity. Thus, we undertook an investigation of both AA metabolism and immunologic response in 20 patients who suffered 40-90% total body surface area (TBSA) burns. Cyclooxygenase (CO:RIA) and lipoxygenase (LO;HPLC det.) metabolites and superoxide (O2 X-) production were measured in stimulated polymorphonuclear cells (PMNL) (A 23187 +/- AA for icosanoid release; phorbol myristate acetate for O2 X-production). Lyso-paf acether (P-LPA) was measured in plasma samples. Ca2+-dependent K+permeability in PMNL was measured by the cell K+ release induced by A 23187. Tc and Tc subsets were determined using monoclonal antibodies (OKT3+, OKT4+ and OKT8+). A biphasic sequential release of the different substances (leukocytic icosanoids and O2 X-) was monitored: increase (approximately 36-48 h after Tl) and decrease (greater than or equal to 72 h after Tl). The increase in AA stimulation was more transient than that of O2 X -. The decline in the release of AA metabolites and O2 X-production was associated with the anergic phase (decrease OKT4+/OKT8+ ratio) and with the clinical outcome of the patients. The decrease in LTB4 and other LO metabolites could explain the impairment of neutrophil chemotaxis. Ca2+ dependent K+ permeability increased early up to 2 or 3 times normal. In order to go further with the mechanism of inhibition of LTB4 and O X-release, the effect of Tl plasma was assayed on normal leukocytes: a 10 min incubation with such plasma was sufficient to abolish LTB4 secretion. A less important inhibition was observed with O2 X-release (-32%) and Ca2+-dependent K+ permeability (-30%). Plasma inhibition seems to be due to a thermolabile factor(s) [protein(s): "suppressive factor(s) of membrane activation "SFMA] which is (are) under active investigation using gel-filtration chromatography and fast protein liquid chromatography (FPLC). Among the SFMAs, certain acute phase proteins could play a key role: i.e., incubation (10 min) of normal PMNL with ceruloplasmin (1 mg/ml) abolished LO products and O X 2-release. PMID- 2989980 TI - Influence of pregnancy and sex steroids on concentration, motor effect and receptor binding of VIP in the rabbit female genital tract. AB - The influence of sex steroid and pregnancy on the tissue concentration, uterine motor effect and receptor binding of VIP has been studied in the female genital tract of pregnant rabbits and oophorectomized rabbits during progesterone and/or oestrogen substitution. The concentration of immunoreactive VIP was high in the vagina and cervix, and lower in the uterine body of both pregnant and non pregnant rabbits. A significant decrease in the VIP concentration (pmol/g wet weight) of the uterine body was observed toward term of pregnancy. The total uterine content of VIP, however, seems unchanged. Treatment of oophorectomized rabbits with ovarian steroids had no effect on the VIP concentration. The sensitivity for and potency of VIP on the relaxation of uterine muscle was significantly higher in oophorectomized rabbits treated with a combination of progesterone and oestrogen than in control rabbits. No difference was observed between non-pregnant and pregnant rabbits. The degradation and binding affinity for 125I-labelled VIP was highest in oophorectomized rabbits substituted with both oestrogen and progesterone. In the pregnant rabbits, the amount of receptors was decreased near term. In conclusion, sex steroids are able to influence the motor effect of VIP at the receptor level, but have no effect on the VIP concentration in the female genital tract. PMID- 2989981 TI - Gonadal dose measurement in diagnostic nuclear medicine using thermoluminescent dosimeters. AB - Instead of using the Medical Internal Radiation Dose Committee (MIRD) concept to calculate the gonadal dose during diagnostic examination using radiopharmaceuticals, measurements of gonadal doses using thermoluminescent dosimeters were studied. The results obtained are comparable with that calculated using the MIRD concept. Five frequently used examinations were investigated, viz., liver, brain, thyroid, bone scintigraphy and cholescintigraphy. The calibration of thermoluminescent dosimeters are fully described, and the experimental techniques are also fully presented. PMID- 2989982 TI - [Biochemical mechanisms of acute inflammation]. PMID- 2989983 TI - [Rare tumor of the external auditory canal. Ceruminoma]. PMID- 2989984 TI - [Clinical and histopathological comments on a case of mixed salivary tumor located in a rare site]. PMID- 2989985 TI - [Spontaneous pneumothorax as the initial symptom of bronchogenic carcinoma]. PMID- 2989986 TI - [The inert gas method in respiratory physiopathology]. AB - The main results obtained, in healthy or unhealthy man, with the inert gas method are gathered together. The major factor at the source of hypoxemia is the uneven distribution of VA/Q ratios. The fall in the partial pressure of oxygen in mixed venous blood amplifies this effect. Only hypoxemia, during muscular exercise, in patients with interstitial lung diseases could be explained, in part, by a decrease in membrane diffusion and a reduction in the transit time of blood in pulmonary capillaries. The differences between the methods used to analyse pulmonary gas exchange are discussed. PMID- 2989987 TI - Protein metabolism in growing pigs fed corn or cassava peel based diets containing graded protein levels. AB - Sixty-four Large White cross Landrace weanling pigs were randomly allotted to eight treatments in a two by four factorial arrangement. The two dietary variables were cassava peel (0 and 40 per cent) and crude protein (20, 15, 10 and 5 per cent). Total serum protein concentration was significantly (P less than 0.01) reduced by protein deficiency and by its interaction with cassava peel. The multiple coefficient of determination (R2) showed that protein intake was the primary factor determining changes in serum protein. R2 values for cyanide intake (independent variable) on serum protein (dependent variable) increased from day 30 to 90 of the trial. Serum urea was increased on the 5 per cent protein diets on days 60 and 90 of the trial. The R2 values for cyanide and protein intake on serum urea concentration increased from day 30 to day 90 of the trial. Serum creatinine increased (P less than 0.05) on the 5 per cent protein diet on day 90 of the trial. The R2 value for the effects of protein intake on serum creatinine was higher than for cyanide intake on days 30 and 90. The results confirm the progressive and pronounced effects of long term cyanide intake on serum nitrogenous metabolites in pigs consuming between 110 and 120 ppm hydrocyanic acid, especially in diets containing 10 per cent or less protein. PMID- 2989988 TI - Pathology of natural rotavirus infection in clinically normal calves. AB - During a longitudinal study of the epidemiology of rotavirus infection in a calf rearing unit, excretion of virus in faeces was detected by enzyme-linked immunosorbent assay in 40 of 48 (83 per cent) unweaned calves aged between three days and five weeks. Fifty per cent of the infected calves had no clinical signs of disease. Enterocytes containing rotavirus antigen and intestinal lesions were found in all of 12 clinically normal calves selected for necropsy between days 1 and 4 of virus excretion. Stunting and fusion of villi, exfoliation, disarrangement and vacuolation of enterocytes and the presence of cuboidal enterocytes were observed in infected calves but not in rotavirus-free control calves. Lesions predominated in the upper small intestine, where rotavirus was most abundant, especially on the first two days of virus excretion. The numbers of enterocytes infected with rotavirus diminished before the lesions resolved. PMID- 2989989 TI - Effects in sheep of high dietary levels of vitamin D3 as measured by circulating concentrations of some vitamin D metabolites and skeletal examination. AB - Housed non-pregnant ewes were given diets supplemented with five, 10, 20, 40 and 80 times the Agricultural Research Council's recommended requirement for vitamin D for 16 weeks. The effects on vitamin D status were assessed by measuring 25 hydroxylated vitamin D metabolites (25-OHD) in plasma at intervals. After 16 weeks, animals on the lowest and two highest vitamin D intakes were killed for histological examination of selected soft tissues and bones. Circulating concentrations of 25-OHD increased with intake and time during the experiment. However the increases were not proportional to intake and only when intakes exceeded 20 times the recommended requirement were concentrations greater than those seen in summer in grazing sheep. There was some evidence for the production of an additional vitamin D metabolite in animals on high vitamin D intakes. No abnormalities were detected in any of the tissues examined histologically. PMID- 2989990 TI - A double-crossover study comparing conventional ventilation with high frequency ventilation in a patient with tracheoesophageal fistula. AB - Respiratory distress, from severe gastric aspiration pneumonitis and abdominal distention in the patient with tracheoesophageal fistula frequently requires mechanical ventilatory support. Bulk flow ventilation can lead to enlargement of the fistulous tract, elevation of gastric intraluminal pressures, raised airway pressures with hemodynamic instability, and retained secretions. We report a case of tracheoesophageal fistula, secondary to perforation of a squamous cell carcinoma of the esophagus, with temporary improvement in gas exchange on high frequency ventilation after failing on a conventional ventilator. The patient initially failed to improve on an Engstrom ventilator (Engstrom-Gambro, Inc., Barrington, IL) at 13 l/minute ventilation. Instituting high frequency jet ventilation with a VS 600 Jet Ventilator (Instrument Development Corporation, Pittsburgh, PA) at initial settings of 35 psi, rate 150, inspiratory time 40%, FiO2 0.8 and 12 cm H2O positive end expiratory pressure (PEEP), provided incremental improvement in gas exchange and oxygenation up to 26 cm H2O PEEP. However, in view of progressive multi-organ failure we terminated the jet ventilation after 48 h and returned the patient to conventional ventilation. We were unable to provide life-sustaining ventilation and oxygenation with either an Engstrom ventilator at 13 l/-minute ventilation or an MA-1 ventilator (Puritan Bennett, Kansas City, MO) at a tidal volume of 800 cc and a ventilator rate of 30. Terminal respiratory failure occurred. Based on the period of improvement using high frequency jet ventilation, we believe this mode of ventilatory support is beneficial in the management of tracheoesophageal fistula. PMID- 2989991 TI - Succinylcholine-induced ventricular arrhythmia during halothane anesthesia. AB - A case of ventricular tachycardia in a healthy child subsequent to onset of a succinylcholine infusion is described. A 9-year-old male patient was scheduled for an adenoidectomy and bilateral tonsillectomies. There was no history of cardiac arrhythmia. Anesthesia was induced by mask with nitrous oxide and halothane. Immediately after the infusion of 20 mg of succinylcholine, the heart rate was 150 beats/min and then the electrocardiograph (ECG) revealed a short run of ventricular tachycardia. The patient was given 1 mg of propranolol, and enflurane and 100% oxygen were administered. Soon after the propranolol was given, normal sinus rhythm returned. Ventricular tachycardia may occur after a single dose of succinylcholine in a healthy anesthetized child without predisposing disease. PMID- 2989992 TI - Responsiveness of cerebral vessels to changes of blood pressure and partial pressure of carbon dioxide after a transient period of cardiac arrest in dogs. AB - Post-ischemic cerebral hypoperfusion supposedly due to constriction of cerebral vessels is considered to be one of the most important factors limiting the recovery of the brain after cerebral ischemia. An experimental study on dogs was carried out to determine the changes in the responsiveness of cerebral vessel to the dilating effects of increase of arterial pressure (AP) and of CO2 inhalation after 3-6 min of cardiac arrest. Responsiveness was measured by the ratio of change in intracranial pressure (ICP) to change in AP (delta ICP/delta AP) and to change in PCO2 (delta ICP/delta PCO2), since in a bony cranium the changes in cerebral vessel diameter are reflected by instantaneous ICP change. delta ICP/delta AP following the administration of intravenous epinephrine was 33%, 43%, 36%, 37% and 16% of pre-ischemic value 1 h, 2 h, 3 h, 4 h and 5 h after cerebral ischemia, respectively. delta ICP/delta PCO2 following 10% CO2 inhalation was 13%, 32%, 55%, 50%, 70% and 75% of pre-ischemic value 1/2 h, 1 h, 2 h, 3 h, 4 h and 5 h after cerebral ischemia, respectively. Normal delta ICP/central venous pressure (delta CVP) was observed during the post-ischemic period, although statistical analysis was not done. From this we concluded: After 3-6 min of cardiac arrest, cerebral arteries constricted for more than 5 h during post-ischemic period. These arteries did not respond well to the dilating effects of increased arterial pressure or CO2 inhalation, but after 3 h their responses to CO2 inhalation returned to their pre-ischemic levels. The intracranial pressure became more or less dependent on CVP during post-ischemic period. PMID- 2989993 TI - A case of attempted suicide by self-hanging. AB - A young man arrived at the accident and emergency department after attempting suicidal self-hanging. He was cyanosed and showed severe respiratory distress. His posture, hyperreflexia and state of consciousness suggested decerebration. A dire prognosis was expected. Four days later he recovered. No residual physical or mental sequelae were found during follow up. The "post-suspension syndrome" in our case was similar to those recorded in several continental reviews. Apart from a case report of a "miraculous deliverance" following an unsuccessful judicial hanging at Oxford in 1650, no other report of this syndrome has been made in the English literature. This syndrome has an excellent prognosis and requires very little active intervention to ensure a favourable outcome. PMID- 2989994 TI - A high performance isolated rat brain preparation. Part I: Operative technique and recipient extracorporeal perfusion. AB - The microsurgical preparation and high level performance with extracorporeal recipient perfusion of a stable, totally isolated rat brain model is presented. Emphasis has been placed on the use of the operating microscope, bipolar cautery and mechanical fixation units to affect a complete ablation of all non-osseous cephalic and vertebral tissues with minimal physiological disturbance of the animal as a whole. The simplified technique of recipient support circulation of the isolated brain organ achieves at least 5 h of tissue viability and function as measured by electrocortical activity, A-Vo2 differences and morphological appearance. To date, no other isolated rodent brain model has been described which is truly anatomically separated from all cephalic tissues. Additionally, other surgical attempts at isolation have required the use of abnormal physiological states, including deep hypothermia and circulatory arrest. PMID- 2989995 TI - A high performance isolated rat brain preparation. Part II. Cerebral blood flow in relation to perfusion pressure and cerebral hypothermia. AB - The effects of changes of perfusion pressure and hypothermia on the cerebral blood flow of a new isolated rat brain preparation have been studied in 7 animals. The cerebral blood flow was extremely sensitive to the mean arterial pressure of perfusion, showing little evidence of autoregulation at low pressure, and only slight evidence in hypertension. The cerebral blood flow was lower in hypothermia. The preparation maintained good electroencephalographic activity throughout, and arteriovenous differences persisted up to 1 h after it became isoelectric. PMID- 2989996 TI - The buffering capacity of crystalloid and colloid resuscitation solutions. AB - The buffering capacities of common colloid and crystalloid resuscitation solutions were compared in vitro. An equal volume of each resuscitation solution was titrated above and below its initial pH with 0.14 N sodium hydroxide or 0.11 N hydrochloric acid. The volume (+/- S.D.) of titration solution necessary to lower the pH one unit (7.1-6.1) in these solutions was less than 0.5 ml for normal saline, less than 0.5 ml for Ringer's lactate, 1.9 +/- 0.1 ml for Plasmalyte-A, 2.0 +/- 0.23 ml for Plasmalyte-R, 8.8 +/- 0.17 ml for human serum albumin (HSA), 45 +/- 2.2 ml for human fresh frozen plasma (FFP), and 50 +/- 6.6 ml for swine FFP. With the method of this in vitro study, human fresh frozen plasma was 25-50 times better as an acid buffer than the crystalloid solutions and approx. 5 times better than human serum albumin (HSA). On an equal volume basis, it was the superior resuscitation solution as a buffer, probably because of combined bicarbonate and protein content. PMID- 2989998 TI - [Myocardial blood flow measurement with argon and a mass spectrometer]. PMID- 2989997 TI - Clinical buffering of metabolic acidosis: problems and a solution. AB - Traditionally sodium bicarbonate has been the buffer of first choice in the treatment of metabolic acidosis. This treatment, however, involves risks of developing a hyperosmolar state, a high sodium concentration in the blood, increased arterial carbon dioxide tension and, as a result of the latter, intracellular and intracerebral acidosis and also cerebral oedema. The buffering effect occurs slowly and as a consequence of this, and of the titration curve of sodium bicarbonate, overcorrection of metabolic acidosis is often seen. Tris buffer was introduced as an alternative and has been claimed to solve most of these problems, but on the other hand it entails a very high risk of peripheral venous thrombosis and thrombophlebitic lesions owing to its local irritative effect. In order to overcome these disadvantages a new mixture of Tris, acetate, bicarbonate and phosphate has been designed. In the studies described it was shown to have an adequate buffering effect and to provide a solution to most of the problems connected with buffering of metabolic acidosis. The new Tris buffer mixture has a buffering effect in blood equivalent to 0.5 mol/l sodium bicarbonate, although its sodium content has been decreased to one-third of pure sodium bicarbonate. Its administration also results in predictable buffering in cerebrospinal fluid and skeletal muscle. In a clinical study it was demonstrated that the new Tris buffer mixture results in sufficient and adequate buffering without significant side-effects. PMID- 2989999 TI - [Clinical usefulness of technetium-99m pyrophosphate and Tl-201 myocardial imaging for the estimation of myocardial infarction--relationship between total serum CPK and the left ventricular function during the acute phase of myocardial infarct]. PMID- 2990001 TI - Sympathetic nervous adjustments in man to simulated weightlessness induced by water immersion. AB - To clarify the role of the sympathetic nervous system to adjust the fluid shift under weightlessness, muscle and skin sympathetic activities were recorded microneurographically in human subjects under simulated weightlessness induced by water immersion up to the levels of the knee, the navel, the breast and the neck. The muscle and skin sympathetic activities were reduced in proportion to rise of immersion level up to the neck. These changes of sympathetic activities were almost concomitant with those of simultaneously recorded soleus electromyograms and heart rate. Reductions of the thigh and the leg circumference were also confirmed by strain gauge plethysmogram recorded under the same experimental condition. Based on these findings, it is concluded that the sympathetic nervous system is suppressed under weightlessness simulated by water immersion. This suppression might depend mainly on the activation of intrathoracic low pressure receptors, due to the fluid shift toward the upper part of the body. The suppression of the sympathetic nervous system seems to be important to compensate the fluid shift under weightlessness. PMID- 2990000 TI - Sensitization-induced sodium influx in airway smooth muscle cells of guinea pigs. AB - Airway smooth muscle (ASM) preparations isolated from Camm-Hartley male guinea pigs were passively in vitro sensitized using Austen's technique. During this sensitization, cells were frequently penetrated with glass microelectrodes (a resistance of 80-90 M omega) for a period of 120 min. Simultaneously, changes in the isometric force of ASM preparations were also monitored. It was found that: administration of serum (1:10 dilution) from sensitized animals caused a depolarization of ASM cells (-52.3 +/- 0.7 mV), achieving a peak at 4 min, followed by slow sustained hyperpolarization reaching a steady state after 15 min; (Resulting resting membrane potential of ASM cells was -68.8 +/- 0.5mV as compared to membrane potential of controls equal to -60.0 +/- 0.7 mV); in the presence of ouabain (10(-5)M), the development of hyperpolarization of ASM cells was completely inhibited; replacement of Na+ with choline chloride or lithium chloride partially inhibited the development of depolarization of ASM cells; in the presence of specific sodium entry blocker, amiloride the sensitization-caused depolarization was abolished; the presence of choline chloride, lithium chloride or amiloride in the experimental medium completely inhibited the contractile response of sensitized tissues to a specific antigen; response of ASM preparations to a specific antigen was absent during depolarization of ASM cells, but was always present during their hyperpolarization. It is concluded that in vitro passive sensitization leads to the interaction of serum specific antibodies with the membrane of airway smooth muscle cells so that permeability of membrane for Na+ is increased (transient depolarization) followed by the activation of electrogenic Na+-pump (sustained hyperpolarization). These events seem to determine the contractile response of sensitized ASM cells to a specific antigen. PMID- 2990003 TI - [Kala-azar--an endemic disease close to Switzerland. Apropos of 2 cases in children observed in Geneva]. PMID- 2990002 TI - [Spontaneous pneumothorax and broncho-pulmonary cancer]. PMID- 2990004 TI - Cramps, spasms and muscle stiffness. AB - Cramp syndromes pose a challenge for neuroscientists. The motor disorders of Isaacs syndrome have been ascribed to peripheral neuropathy, and sometimes there is ample supporting evidence of neuropathy. However, signs of overt neuropathy are found in a minority of cases and the essential findings (carpal and pedal spasm, pseudomyotonia and myokymia) may arise from abnormal excitability of the perikaryon because similar manifestations are seen in tetany and multiple sclerosis. The Moersch-Woltman (stiffman) syndrome differs from Isaacs' syndrome in essential characteristics. Hyperventilation syndromes may mimic either simple cramps, the Isaacs syndrome, the Moersch-Woltman syndrome, or the Foley and Denny Brown syndrome of benign fasciculation and cramps. New approaches are needed to define the etiology and pathogenesis of these neurogenic disorders because the results of peripheral nerve block and spinal anesthesia have not been consistent in cases of typical Isaacs syndrome. Occupational cramps can be regarded as a form of action dystonia but that statement is a clue, not an "explanation". Myopathic disorders are only rarely a cause of cramp syndromes. In the glycogen storage disorders, the chemical basis of the cramp is still unproven. Whether myoadenylate deaminase is a cause of cramps is debated. PMID- 2990006 TI - [Painful legs and unstable toes]. AB - Five patients presented with the painful legs and moving toes syndrome as defined by Spillane et al. (1971). A peripheral nervous disorder was present in 4 of these cases while the fifth patient showed evidence of a central lesion. Results of clinical and electrophysiologic investigations in the 4 peripheral cases, are compared with data from the literature. PMID- 2990005 TI - [Adult disclosure of a case of familial adrenoleukodystrophy]. AB - We report a case of adult adrenoleukodystrophy. The patient, originating from North Africa, had no clinical history until age 23. The first disorder, a spastic paraparesis, occurred after a 5 days coma following a cranial traumatism. The course of the disease was then progressive remitting and the patient died at age 30. Total duration was 7 years. The diagnosis of A.L.D. was ascertained by the determination of excessive very long chain fatty acid level in plasma. Endocrinological study revealed adrenal insufficiency but darkening of the skin was masked by racial pigmentation. The patient was the oldest of 10 children. 3 of the brothers died of childhood A.L.D. around the age of 12 after a progressive 2 years course. The mother and 2 asymptomatic brothers have increased plasma levels of hexacosanoic acid and A.C.T.H. The particularity of this adult case is the lack of spinal demyelination. Pathological studies revealed a widespread cerebral demyelination with an inflammatory process. The features of this case are compared to those of the 13 other reported cases of adult A.L.D. PMID- 2990007 TI - Some epidemiologic aspects of yaws in the Ivory Coast. AB - Because of the prevailing high levels of yaws, a mass treatment campaign was conducted in the Ivory Coast from 1956 to 1970. In 1970, at the end of the mass campaign, less than 30,000 cases of yaws were reported. After 1970, a program of detection and treatment of remaining foci of infection was carried out to control the disease. The number of cases reported annually continued to decline until 1974, when the number of reported cases began to rise. Since 1980, the annual reported incidence of yaws has declined. It is concluded that the mass campaign of 1956-1970 reduced the endemicity of yaws in the Ivory Coast. However, significant foci of the disease remain, and efforts to detect and treat yaws must be continued to prevent a resurgence of this infection in the Ivory Coast. PMID- 2990008 TI - [Endemic tuberculosis in the Cluj District 1980-1983]. PMID- 2990009 TI - [Thoracoscopy--a diagnostic and treatment method in pneumology]. PMID- 2990010 TI - [Chronic obstructive bronchopneumonia associated with diffuse interstitial fibrosis]. PMID- 2990011 TI - [Electronic technics in exploring the chest]. PMID- 2990012 TI - [Pulmonary tuberculosis associated with hepatic lesions]. PMID- 2990013 TI - [Masks of tuberculous meningoencephalitis in the adult (a case report)]. PMID- 2990015 TI - [Current directions in the antibiotic therapy of acute nontuberculous infectious pneumonias in adults]. PMID- 2990016 TI - [Acquired immune deficiencies]. PMID- 2990014 TI - [Case of MacLeod's syndrome]. PMID- 2990017 TI - [Insulinoma today]. PMID- 2990018 TI - [Sensori-motor neuropathy in rheumatoid polyarthritis. Anatomo-clinical study]. PMID- 2990019 TI - Evaluation of pituitary-adrenal function after pituitary surgery. AB - A short overnight metyrapone test and a 30-min ACTH test were performed in ten patients after pituitary surgery. At the initial testing 2 weeks after surgery the 30-min ACTH test was abnormal in two patients while normal increases in s cortisol were observed in eight patients. These eight patients also had normal responses to ACTH when re-tested after 6-19 months. The short metyrapone test was a much more sensitive indicator of functional disturbances in pituitary/adrenal function. Two weeks after surgery the test was abnormal in seven of the patients. After 6-19 months some normalization of the short metyrapone test had occurred, probably due to disappearance of postoperative oedema and haematoma. However, the test was still abnormal in three patients having normal responses during the 30 min ACTH test. It is suggested that both tests are performed in such patients to specify a high risk group, i.e. both tests abnormal, and a low risk group with a normal 30-min ACTH test but subnormal responses during the short metyrapone test. This would be of help in the decision on cortisol supplementation and offer a high degree of safety for the patients. PMID- 2990020 TI - The calcium ion activity and the standardized excretion rate of calcium in urine of healthy adults. AB - A standardized protocol is described for the study of the calcium excretion in urine. After 12 h of fasting, urine is collected during 4 h with a water load of 10 ml per kg body weight. Urine is also collected during the following 20 h period on the habitual water and calcium intake. Reference values for 48 healthy adults are given as 0.25 and 0.75 quantiles. The measured activity of calcium ions (Ca2+) in urine is 0.09-0.27 mmol/kg for the 4 h period, 0.34 to 0.52 mmol/kg for the 20 h period; pH values are 5.61-6.43 (4 h) and 5.46 to 6.04 (20 h). The concentrations of total calcium are 0.67-2.05 mmol/l (4 h) and 3.16 to 4.94 mmol/l (20 h). The value for the excretion rate of calcium (standardized to a creatinine clearance of 100 ml/min) is 1.30-3.24 mumol/min for the 4 h period and 3.06-4.88 mumol/min for the 20 h period, with no significant difference between the results for men and women. The relationship between the Ca2+ activity and pH was studied in urine titrated with HC1 or NaOH. In all urine the Ca2+ activity falls with increasing pH in a typical biphasic manner. This indicates the need for simultaneous measurement of the pH in order to interpret data for the Ca2+ activity in urine. PMID- 2990021 TI - The inherited basis for venous thrombosis. The thrombohaemorrhagic balance. PMID- 2990022 TI - Combined factor V and factor VIII deficiency with normal protein C and protein C inhibitor. A family study. AB - Combined deficiency of factor V and factor VIII, a rare bleeding disorder, is reported in a Syrian family. 2 siblings, 10 and 6 yr old are affected. They had mild bleeding manifestations. Their prothrombin time and partial thromboplastin time were prolonged, but thrombin time was normal. Both had low levels of factor V, (6% and 7%), factor VIII, (both 10%) factor VCAg (both 6%) and factor VIII CAg, (6% and 4%). All members of this family had normal levels of factor VIIIR:Ag, protein C, antigen and protein C inhibitor. The normal levels of protein C and protein C inhibitor in the 2 affected family members indicate that the combined deficiency of factors V and VIII has nothing to do with protein C. This contrasts with previous reports that deficiency of protein C inhibitor is the cause of combined factor V and factor VIII deficiency. The probable mode of inheritance of this defect is discussed. PMID- 2990023 TI - Occurrence of HTLV-I antibodies in Danish patients with cutaneous T-cell lymphoma. AB - 10 of 68 CTCL (cutaneous T-cell lymphoma) patients without features of ATLL had antibodies against HTLV-I (human T-cell leukaemia virus, type I). The titre of antibody in these positive patients was generally much lower than that seen in cases of ATLL (adult T-cell leukaemia/lymphoma); geometric mean of 80 for CTCL vs. 8000 for Caribbean ATLL. The presence of HTLV-I antibody was unrelated to clinical remission, relapse, or stages of the disease, and some positives were detected in the earliest phases of mycosis fungoides. Among controls and normal donors between the ages of 40 and 65, only 1 of 36 and 3 of 113, respectively, had low titre antibodies to HTLV-I in their sera. Only 5 of 354 Danish normal donors of all ages had antibody, which was identical to the rate in over 2000 US normal donors. In negative control experiments, these antibodies were unreactive with bovine leukaemia virus. These data suggest that HTLV-I or a related retrovirus crossreactive with HTLV-I occurs in a low percentage of the Danish population and patients with CTCL have such antibodies at an increased rate, but less than the rate seen for ATLL (greater than 90%). PMID- 2990024 TI - [Apolipoprotein E]. PMID- 2990025 TI - [Enkephalin-noradrenaline interactions in synaptic transmission]. PMID- 2990026 TI - [Regulation of contractile proteins in platelets--phosphorylation-myosin light chain theory]. PMID- 2990027 TI - Studies of the putative transforming protein of the type I human T-cell leukemia virus. AB - The putative transforming protein of the type I human T-cell leukemia virus (HTLV 1) is a 40-kilodalton protein encoded by the X region and is termed p40XI. On the basis of both subcellular fractionation techniques and immunocytochemical analysis, it is now shown that p40XI is a nuclear protein with a relatively short half-life (120 minutes). It is synthesized de novo in considerable quantities in a human T-cell line infected with and transformed by the virus in vitro, and it is not packaged in detectable amounts in the extracellular virus. PMID- 2990028 TI - A transcriptional activator protein encoded by the x-lor region of the human T cell leukemia virus. AB - Human T-cell leukemia viruses type I and II (HTLV-I and -II) exhibit several features characteristic of this retroviral family: the presence of an x-lor gene encoding a nuclear protein, transformation properties suggesting the involvement of a virus-associated trans-acting factor, and transcriptional trans-activation of the long terminal repeat (LTR) in infected cells. In the study described here the HTL x-lor products, in the absence of other viral proteins, were able to activate gene expression in trans directed by HTLV LTR. The regulation of the expression of particular genes in trans by HTLV x-lor products suggests that they play a role in viral replication and possibly in transformation of T lymphocytes. PMID- 2990029 TI - Morphine-induced delay of normal cell death in the avian ciliary ganglion. AB - Repeated administration of morphine in increasing doses delayed normal cell death in the ciliary ganglion of the chick embryo; the effect was completely blocked by naloxone. Survival of spinal motoneurons was not affected. Morphine also inhibited potassium-stimulated synthesis of acetylcholine in ganglion cells cultured with muscle, suggesting that morphine can influence neurotransmission. Morphine's effect on cell death may be due to an inhibition of transmission at the neuromuscular junction, but opiates may also directly affect cell death. Although it is now known whether the endogenous opiates in the ciliary ganglion influence neuronal survival during embryogenesis, exogenous opiates can affect normal cell death in the autonomic nervous system. PMID- 2990030 TI - The unusual varl gene of yeast mitochondrial DNA. AB - The var1 gene specifies the only mitochondrial ribosomal protein known to be encoded by yeast mitochondrial DNA. The gene is unusual in that its base composition is nearly 90 percent adenine plus thymine. It and its expression product show a strain-dependent variation in size of up to 7 percent; this variation does not detectably interfere with function. Furthermore, var1 is an expandable gene that participates in a novel recombinational event resembling gene conversion whereby shorter alleles are preferentially converted to longer ones. The remarkable features of var1 indicate that it may have evolved by a mechanism analogous to exon shuffling, although no introns are actually present. PMID- 2990031 TI - Expression of the pX gene of HTLV-I: general splicing mechanism in the HTLV family. AB - Human T-cell leukemia virus type I (HTLV-I) is an etiological agent of adult T cell leukemia. A viral gene pX encodes for p40X and it has been proposed that this protein trans-activates the viral long terminal repeat and possibly some cellular genes; this activation may be associated with T-cell transformation. The mechanism of pX gene expression and the primary structure of p40X are now reported. Two-step splicing generates the 2.1-kilobase pX mRNA; the initiator methionine for env becomes part of the pX protein. These splicing signals are conserved among all members of the HTLV family except for the acquired immune deficiency syndrome-associated viruses. PMID- 2990032 TI - HTLV x-gene product: requirement for the env methionine initiation codon. AB - The human T-cell leukemia viruses (HTLV) are replication-competent retroviruses whose genomes contain gag, pol, and env genes as well as a fourth gene, termed x, which is believed to be the transforming gene of HTLV. The product of the x gene is now shown to be encoded by a 2.1-kilobase messenger RNA derived by splicing of at least two introns. By means of S1 nuclease mapping of this RNA and nucleic acid sequence analysis of a complementary DNA clone, the complete primary structure of the x-gene product has been determined. It is encoded by sequences containing the env initiation codon and one nucleotide of the next codon spliced to the major open reading frame of the HTLV-I and HTLV-II x gene. PMID- 2990033 TI - Imaging elemental distribution and ion transport in cultured cells with ion microscopy. AB - Both elemental distribution and ion transport in cultured cells have been imaged by ion microscopy. Morphological and chemical information was obtained with a spatial resolution of approximately 0.5 micron for sodium, potassium, calcium, and magnesium in freeze-fixed, cryofractured, and freeze-dried normal rat kidney cells and Chinese hamster ovary cells. Ion transport was successfully demonstrated by imaging Na+-K+ fluxes after the inhibition of Na+- and K+ dependent adenosine triphosphatase with ouabain. This method allows measurements of elemental (isotopic) distribution to be related to cell morphology, thereby providing the means for studying ion distribution and ion transport under different physiological, pathological, and toxicological conditions in cell culture systems. PMID- 2990034 TI - Malignant transformation of erythroid cells in vivo by introduction of a nonreplicating retrovirus vector. AB - DNA from a replication-defective spleen focus-forming virus (SFFV) was reconstructed and transfected into psi-2 cells containing a packaging-defective mutant of Moloney murine leukemia virus. Replication-incompetent retrovirus particles (helper virus-free containing genomes that express the transforming envelope gene of SFFV (gp52) transformed bone marrow cells in vitro and, after direct intravenous introduction of the vector, induced malignant erythroid disease in vivo. Disease induction was dependent on prior treatment of mice with phenylhydrazine, which probably increased the availability of erythroid target cells. Since there was no evidence of virus particle expression in mice with malignant disease, this study demonstrates the acute oncogenic potential of a limited number of erythroid cells expressing SFFV gp52. Direct inoculation of animals with nonreplicating retroviral vectors containing transforming genes may be useful in study the oncogenic effects of such genes. PMID- 2990035 TI - The granulocyte-macrophage colony-stimulating factors. AB - The granulocyte-macrophage colony-stimulating factors are well-characterized specific glycoproteins that interact to control the production, differentiation, and function of two related white cell populations of the blood, the granulocytes and monocyte-macrophages. Widely produced in the body, these regulators probably play an important role in resistance to infections. The proliferation of myeloid leukemia cells remains dependent on stimulation by colony-stimulating factors, although one of them also has the ability to suppress leukemic populations by inducing terminal differentiation. PMID- 2990036 TI - More about the HTLV's and how they act. PMID- 2990037 TI - The x gene is essential for HTLV replication. AB - The human T-cell leukemia viruses (HTLV) are associated with T-cell malignancies in man and will transform normal human T cells in vitro. The mechanism of malignant transformation by HTLV is unknown but appears to be distinct from that of other classes of retroviruses, which induce malignant transformation through viral or cellular oncogenes. Recently a new gene, termed x, was identified in HTLV. This gene has been hypothesized to be the transforming gene of HTLV because of its conservation within the HTLV class of retroviruses. By in vitro mutagenesis of the HTLV-II x gene, it is now demonstrated that the presence of a functional x gene product is necessary for efficient HTLV transcription. Therefore, these studies provide direct evidence for an important function of the x gene in HTLV replication. The functional analogies between the x gene and transcriptional regulatory genes of some DNA viruses suggest that these viruses share similar mechanisms for cellular transformation. PMID- 2990038 TI - Reexamination of glucose-6-phosphatase activity in the brain in vivo: no evidence for a futile cycle. AB - Glucose-6-phosphatase activity in the rat brain in vivo was estimated by measuring the differential loss of tritium and carbon-14 from the glucose pool labeled by a mixture of [2-3H]glucose and [U-14C]glucose. The results provide no evidence of significant dephosphorylation of glucose-6-phosphate and do not support the hypothesis of a futile cycle involving glucose-6-phosphatase activity in the brain. PMID- 2990039 TI - A role for glycosylation of the alpha subunit in transduction of biological signal in glycoprotein hormones. AB - The biological properties of recombinants of glycoprotein hormones in which the alpha and beta subunits were differentially deglycosylated have been investigated. Specific deglycosylation of the alpha subunit generated a recombinant that had more receptor-binding activity but did not produce hormone response in the target cells. The deglycosylated alpha + beta recombinant was also an antagonist of the action of the native hormone. Thus, the carbohydrates in the alpha subunit play a dominant role in the transduction of the hormone signal into the cell. PMID- 2990040 TI - Trans-activator gene of human T-lymphotropic virus type III (HTLV-III). AB - Human T-lymphotropic virus type III (HTLV-III) encodes a trans-acting factor that activates the expression of genes linked to the HTLV-III long terminal repeat. By functional mapping of complementary DNA transcripts of viral messenger RNA's the major functional domain of the gene encoding this factor was localized to a region immediately before the env gene of the virus, a region previously thought to be noncoding. This newly identified gene consists of three exons, and its transcription into messenger RNA involves two splicing events bringing together sequences from the 5' part (287 base pairs), middle (268 base pairs), and 3'part (1258 base pairs) of the HTLV-III genome. A similar messenger RNA with a truncated second exon (70 base pairs) does not encode a trans-acting function. It is proposed that this second messenger RNA is the transcript of a gene (3'-orf) located after the env gene. Messenger RNA's were also identified for the env and gag-pol genes of HTLV-III. PMID- 2990041 TI - Location of the trans-activating region on the genome of human T-cell lymphotropic virus type III. AB - The retrovirus involved in acquired immune deficiency syndrome (HTLV-III/LAV) contains a region that is necessary for stimulation of gene expression directed by the viral long terminal repeat. This region is located between nucleotides 5365 and 5607, immediately 5' to the envelope gene. A doubly-spliced message containing this region could encode an 86-amino acid protein with structural features similar to those of nucleic acid-binding proteins. PMID- 2990042 TI - An RNA processing activity that debranches RNA lariats. AB - The excised introns of pre-messenger RNA's (pre-mRNA's) and intron-containing splicing intermediates are in a lariat configuration in which the 5' end of the intron is covalently joined by a 2',5'-phosphodiester bond to a specific adenosine residue near the 3' end of the intron. A 2',5'-phosphodiesterase activity in HeLa cell extracts has been detected that debranches RNA lariats, converting them to linear RNA molecules by specific cleavage of the 2',5' phosphodiester bond. This lariat debranching activity is distinct from previously reported 2',5'-phosphodiesterases with regard to its biochemical and substrate requirements as well as its stringent cleavage specificity. The debranching activity is observed only if the RNA lariats generated during in vitro processing are deproteinized and added back to the extract. These results suggest that during the normal in vitro splicing reaction the 2',5'-phosphodiester bond of RNA lariats is protected from cleavage by the lariat debranching activity. PMID- 2990043 TI - Putting the human genome on the map. PMID- 2990044 TI - A large deletion within the T-cell receptor beta-chain gene complex in New Zealand white mice. AB - The T-cell receptor beta-chain gene complex contains a duplication of D beta, J beta, and C beta gene segments in mice and man. When DNA from many inbred strains of mice was screened an unusual allele of the beta locus was identified in New Zealand White (NZW) mice. This allele is distinguished by the deletion of an 8.8 kilobase segment of DNA containing C beta 1, D beta 2 and the J beta 2 cluster. Despite the fact that all NZW T-cell receptors must be derived from a single set of beta-chain gene segments, this strain has functional T cells and is phenotypically normal. This deletion of T-cell receptor beta-chain segments occurs in a strain known to contribute to lupus-like autoimmune disease. PMID- 2990045 TI - Discovery of a predicted DNA knot substantiates a model for site-specific recombination. AB - The mechanism of site-specific genetic recombination mediated by Tn3 resolvase has been investigated by a topological approach. Extrapolation of a detailed model of synapsis and strand exchange predicts the formation of an additional DNA product with a specific knotted structure. Two-dimensional gel electrophoresis of DNA reacted in vitro revealed a product, about 0.1 percent of the total, with the appropriate mobility. A technique for determining DNA topology by electron microscopy was improved such that less than a nanogram of DNA was required. The structure of the knot was as predicted, providing strong evidence for the model and showing the power of the topological method. PMID- 2990046 TI - A general method for saturation mutagenesis of cloned DNA fragments. AB - A new procedure for generating and isolating random single-base substitutions in cloned DNA fragments is presented. The mutations are generated by treatment of single-stranded DNA with various chemicals, followed by the synthesis of the complementary strand with reverse transcriptase. Misincorporation frequently occurs when the enzyme encounters a damaged base in the mutagenized template DNA. The resulting duplex DNA fragments containing random single-base substitutions are cloned, amplified as a population, and isolated from wild-type DNA by preparative denaturing gradient gel electrophoresis. The physical separation of mutant DNA fragments makes it possible to isolate and characterize large numbers of site-directed single-base substitutions in the absence of a phenotypic selection. This procedure should be generally applicable to the fine-structure genetic analysis of regulatory and protein-coding sequences. PMID- 2990047 TI - Cyclic AMP regulation of eukaryotic gene transcription by two discrete molecular mechanisms. AB - In experiments designed to study the mechanism by which peptide hormones binding to their plasma membrane receptors stimulate the expression of specific genes, the transcription of two neuroendocrine genes, prolactin and growth hormone, was analyzed in a rat pituitary cell line. The results showed that cyclic adenosine monophosphate (cyclic AMP) stimulates the transcription of discrete subsets of eukaryotic genes by at least two independent molecular mechanisms. Cyclic AMP stimulated growth hormone gene transcription and phosphorylation of a 19,000 dalton nuclear protein; this appears to reflect direct nuclear actions of the cyclic AMP-dependent protein kinase. In contrast, the stimulation by cyclic AMP of prolactin gene transcription appears to reflect activation of a discrete calcium-dependent event. PMID- 2990048 TI - Constitutive and conditional suppression of exogenous and endogenous genes by anti-sense RNA. AB - Plasmid DNA directing transcription of the noncoding (anti-sense) DNA strand can specifically inhibit the expression of several test genes as well as normal, endogenous genes. The anti-sense plasmid constructions can be introduced into eukaryotic cells by transfection or microinjection and function in both transient and stable transformation assays. Anti-sense transcripts complementary to as little as 52 bases of 5' untranslated target gene mRNA specifically suppress gene activity as well as, or more efficiently than, anti-sense transcripts directed against the protein coding domain alone. Conditional anti-sense inhibition is accomplished with the use of hormone-inducible promoter sequences. Suppression of endogenous actin gene activity by anti-sense RNA is detected as a decrease in growth rate and as a reduction in the number of actin microfilament cables. These observations suggest that anti-sense RNA may be generally useful for suppressing the expression of specific genes in vivo and may be a potential molecular alternative to classical genetic analysis. PMID- 2990049 TI - NIMH to reorganize extramural research. PMID- 2990050 TI - Vasopressin-stimulated release of atriopeptin: endocrine antagonists in fluid homeostasis. AB - Administration of pharmacological doses of arginine-vasopressin, related peptides, and other pressor agents induced a profound release of atriopeptin immunoreactivity into the circulation. The stimulated release of atriopeptin apparently was related to increased arterial blood pressure. Neither the nonpressor vasopressin analog 1-deamino-D-Arg8-vasopressin nor arginine vasopressin in the presence of a specific pressor antagonist caused atriopeptin to be released into the circulation. Urine output was correlated with the level of atriopeptin released. Physiological levels of arginine-vasopressin suppress diuresis and produced vasoconstriction. Pharmacological levels of the hormone stimulated the cardiac endocrine system to release atriopeptin, which may cause diuresis and vasodilation to physiologically antagonize the effects of vasopressin. PMID- 2990052 TI - [An experimental study on cartilage and the morphogenesis induced by implanted demineralized dentin matrix]. PMID- 2990051 TI - cis- and trans-acting transcriptional regulation of visna virus. AB - Visna virus is a pathogenic lentivirus of sheep that is related to human T-cell lymphotropic virus type III (HTLV-III), the probable etiologic agent of the acquired immune deficiency syndrome (AIDS). The transcriptional activity of visna virus promoter and enhancer sequences was studied by means of an assay based on the transient expression of the bacterial gene chloramphenicol acetyltransferase (CAT). The results suggest that the high level of expression of visna virus is due in part to cis-acting enhancer sequences that give the viral promoter a high level of transcriptional activity. In addition, the rate of transcription from the visna virus promoter situated in a plasmid expressing the CAT gene was much greater in infected than uninfected cells. This phenomenon of trans-acting transcriptional activation may involve either virally or cellularly encoded factors. PMID- 2990053 TI - Thymic epithelial neoplasms. AB - We studied 57 tumors of the thymus seen at Vanderbilt University Medical Center between 1957 and 1982. By applying current diagnostic criteria, the classification of 13 tumors was changed. There were 45 epithelial tumors of thymus, including 32 thymomas, two thymic carcinomas, eight carcinoids, and three germinomas. The gross and microscopic features of these neoplasms overlap each other and several other tumors, but with knowledge of the variations it was usually possible to establish a diagnosis with light microscopy. In more difficult cases, electron microscopy was the most useful diagnostic method. New developments in immunologic staining were also of value. The diagnosis of thymoma was used for circumscribed and invasive neoplasms of thymic epithelium without anaplasia. Cytologically malignant thymic epithelial tumors were classified as thymic carcinoma. Thymic carcinomas were more aggressive than thymomas and were not associated with syndromes such as myasthenia gravis or anemia. Thymic carcinoids were frequently aggressive, and several of them produced corticotrophin or calcitonin. Thymic germinomas were histologically identical to testicular seminoma, but may be confused with thymoma because of their lymphocytic infiltrate. Invasive germinomas were radiosensitive. PMID- 2990054 TI - Acute renal failure and jaundice associated with Epstein-Barr virus infection. AB - We have described a case of Epstein-Barr virus infection associated with acute renal failure and jaundice. Atypical lymphocytes and absolute lymphocytosis were absent, and the patient recovered completely. PMID- 2990055 TI - Elastosis in malignant tumours. AB - Elastosis is common in infiltrating ductal and lobular carcinomas of the breast, occurring in approximately 90% of cases. It is also well described in some benign lesions of the breast and tumours of the salivary gland. Reports of venous elastosis in association with large-bowel carcinomas are rare. We describe elastosis in single cases of prostatic, gastric, bronchiolar-alveolar and cervical carcinoma. PMID- 2990056 TI - Identification of five new variable restriction sites in HSV-1 DNA. AB - Restriction endonuclease analysis of herpes simplex virus type 1 (HSV-1) DNA has shown that individual strains differ from one another. The differences can be recognized by the presence or absence of cleavage sites due to base changes, small insertions and occasional deletions. In a study in which 13 HSV-1 isolates from South Africa were analysed, a total of 5 new variable restriction sites were identified using the restriction endonucleases Hind III, Eco RI and Bgl II. PMID- 2990057 TI - In-vitro studies with SF 86-327, a new orally active allylamine derivative. AB - SF 86-327 (Sandoz Forschungsinstitut) is an orally active allylamine derivative related to naftifine. The antifungal activity of SF 86-327 was compared in vitro with those of naftifine, ketoconazole, and itraconazole (R 51,211, Janssen Pharmaceutica) by agar dilution. 120 fungal isolates were tested. Also, the antifungal activities of SF 86-327 and naftifine against 18 dimorphic pathogens were assayed in vitro by broth dilution. Results of these studies support claims that SF 86-327 is a broad spectrum antifungal agent. Results of the broth dilution studies also revealed that SF 86-327 was both fungistatic and fungicidal in vitro for isolates of Blastomyces dermatitidis, Histoplasma capsulatum and Sporothrix schenckii at concentrations as low as 0.05 micrograms ml-1 (18 isolates tested, MIC90 = 0.39 micrograms ml-1, MFC90 = 12.5 micrograms ml-1). PMID- 2990059 TI - [Effects of aluminium impurities in the 99mTc generator eluate on radiopharmaceutical behavior]. PMID- 2990058 TI - Management of nonfunctioning islet cell tumors of the pancreas. AB - Eleven patients with nonfunctioning islet cell tumor of the pancreas were examined. Since no hypersecretory syndromes were noted, patients' complaints related to enlarging pancreatic masses. Pancreatoduodenectomy was the most commonly performed procedure, with long-term survival of 77 per cent and 62 per cent at two and five years, respectively. Immunohistochemical stains for various polypeptides were positive in two of four tumors tested. Surgical therapy remains the most effective treatment for resectable lesions, with prolonged survival possible in most patients. PMID- 2990060 TI - [Stoma care is something different than putting a bag on the abdomen]. PMID- 2990061 TI - [Dilated cardiomyopathy. An analysis of the viral-immunological theory]. PMID- 2990062 TI - Benzodiazepines inhibit human platelet activation: comparison of the mechanism of antiplatelet actions of flurazepam and diazepam. AB - These studies were undertaken to examine the effects and the mechanism of action of flurazepam and diazepam on human platelet activation. One minute preincubation with flurazepam (3-300 microM) or diazepam (3-300 microM) inhibited platelet aggregation, serotonin secretion and prostaglandin synthesis induced by ADP (1-5 microM), epinephrine (1-5 microM), and arachidonic acid (600-1000 microM). However, 357% higher concentration of diazepam (265 microM) as compared to flurazepam (58 microM), was required to inhibit arachidonic acid induced production of malondialdehyde (MDA) by 50%. In addition, flurazepam and not diazepam inhibited the release of arachidonic acid from platelet phospholipids in a concentration dependent manner. In other experiments flurazepam but not diazepam also blocked aggregation and secretion induced by U46619 (2 microM), a stable analog of prostaglandin H2. Platelet aggregation and serotonin secretion induced by collagen (40-300 micrograms/ml) was inhibited by flurazepam with an IC 50 of 153 microM and 136 microM respectively, whereas higher than 300 microM diazepam was required to inhibit collagen-induced aggregation and secretion by 50%. Flurazepam and diazepam both exhibited their most potent antiplatelet effects against phospholipase C-induced aggregation which is mediated by prostaglandin-independent mechanisms. Only 15 microM and 11 microM flurazepam and 31 microM and 27 microM diazepam were needed to inhibit PLC-induced aggregation and secretion of serotonin by 50% respectively. Effects of these benzodiazepines on platelet cyclic AMP and cyclic GMP were also examined. Neither flurazepam nor diazepam caused any significant change in cyclic AMP or cyclic GMP levels in platelets. These findings suggest that: (a) flurazepam, as compared to diazepam, is 106% - 357% more effective in inhibiting platelet aggregation and serotonin secretion induced by arachidonic acid, collagen and phospholipase C; (b) flurazepam inhibits platelet activation by inhibiting the release of arachidonic acid, its conversion into prostaglandins and by blocking the action of prostaglandins on platelets; (c) diazepam does not inhibit thrombin-induced release of arachidonic acid, conversion of exogenously added arachidonic acid into MDA, or the action of prostaglandins; (d) both flurazepam and diazepam inhibit PLC-mediated activation of platelets; and (e) neither diazepam nor flurazepam achieve their antiplatelet actions by affecting platelet cyclic nucleotide levels. PMID- 2990063 TI - Hydrolysis of polyphosphoinositides in human platelets. AB - Phospholipase C was purified 110 fold from human platelets. The activity of the enzyme was totally dependent upon Ca2+. The activity of the enzyme was markedly enhanced in the presence of arachidonic acid and was strongly inhibited by aminoglycoside antibiotics. The enzyme hydrolyzed endogenous polyphosphoinositides in addition to PI in Ca2+ dependent manner, suggesting the involvement of this enzyme in stimulus-linked rapid hydrolysis of polyphosphoinositides in platelets. The stimulation by thrombin of 32P-labelled human platelets induced about 30% decrease in 32P-TPI and about 220% increase in 32P-PA at the first 10 sec. The degree of hydrolysis of TPI was dependent upon the amount of agonist and it was not affected by the extracellular concentration of Ca2+. The changes in 32P-phospholipids in thrombin-stimulated platelets in the absence of Ca2+ were inhibited in a dose dependent manner by preincubation with relatively higher amount of quin 2 AM. The inhibition was completely overcome by an addition of CaCl2 to the suspending buffer. By such treatment in the absence of extracellular Ca2+, the intracellular Ca2+ concentration was significantly lowered below the basal level (less than 100 nM). Those observations suggest that TPI breakdown in thrombin-stimulated platelets is primary mediated by the agonist receptor coupling and requires at least the basal level of intracellular Ca2+. PMID- 2990064 TI - [Metronidazole (Flagyl) and polyneuropathy]. PMID- 2990065 TI - [Extragonadal germ cell cancer in the mediastinum]. PMID- 2990066 TI - [Susceptibility of birds other than chickens to infectious laryngotracheitis]. AB - Susceptibility to infectious laryngotracheitis virus was studied in peafowl (Pavo cristatus), various species of pheasant (Phasianus colchicus, Lophura swinhoeii, Lophophorus impejanus), guinea-fowl (Numida meleagris), canaries (Serinus canaria), budgerigars (Melopsittacus undulatus) and Japanese quail (Coturnix coturnic japonica). Apart from clinical observations, experiments were evaluated in terms of histopathology, immunofluorescence, serology and recovery of virus. Only peafowl and pheasants were found to be susceptible, pheasants responding more strongly than chickens to ocular vaccination and intratracheal inoculation. The other species were found to be refractory. PMID- 2990067 TI - Detection of antibodies to cell surface antigens by a simplified cellular ELISA (CELISA). AB - An improved method for screening human hybridoma antibodies to cell surface antigens is described. The following modifications have been developed: rapid expansion of desired screening cell types by EBV transformation; use of only 5 X 10(4) cells/well; elimination of the need for glutaraldehyde fixation; elimination of the requirement for PLL to attach cells to microplates; preparation of a large number of plates which can be stored at 4 degrees for 3 months; Protein A-peroxidase ELISA assay yielding excellent replicates, low background "noise", and high OD readings for positive wells. The techniques we have developed should greatly simplify and shorten the assay procedures for detecting human antibodies to a variety of cell surface antigens. PMID- 2990068 TI - Regulation of the microfilament system in normal and polyoma virus transformed cultured (BHK) cells. AB - The content and state of actin in baby hamster kidney (BHK) cells before and after transformation with polyoma virus were examined by deoxyribonuclease assay and gel electrophoresis followed by dye elution. The actin content of the transformed cells, relative to total cell protein, was lower than that of the normal cells by 30-50%. In both the normal and transformed cells the greater part of the total actin was found on lysis to be in the monomeric state. Cytoplasmic and membrane fractions of the two cell lines were, in qualitative terms, very similar in their protein compositions. The plasma membrane isolated from the transformed cells was richer in actin than that from the untransformed, and both membrane fractions contained proteins corresponding to myosin, filamin and alpha actinin on SDS-polyacrylamide gels. The cell extract from both the normal and transformed lines formed an actin-based gel on incubation at 30 degrees C, although the amount of the cross-linked actin was much smaller in the latter. This was a consequence not only of the lower concentration of total actin in the cell, but also, presumably, of a gross relative deficiency in the concentration or activity of filament cross-linking protein(s) in the cytoplasm. Thus, small aliquots of cytoplasmic fractions from transformed cells, when added to an excess of exogenous F-actin, were able to cross-link the filaments to a much smaller extent than those from the normal cells. A similar range of proteins was found to be associated with the actin gels formed from both cell extracts. One conspicuous difference was that a species migrating in SDS-gel electrophoresis as a doublet with a subunit molecular weight of about 58,000, and tentatively identified as intermediate filament protein, was replaced in the transformed cells by a single band. Filament cross-linking activity of the cytoplasmic fractions was enhanced by addition of Triton extracts of crude membranes, although the latter were not capable of cross-linking exogenous F-actin on their own. The effect of Triton extracts was much greater in the case of membranes from the transformed cells. The cytoplasmic fractions of BHK cells contain capping protein(s) and/or complexes of such proteins with actin; these reveal themselves by the propensity of the extracts to nucleate polymerization of exogenous G-actin. This activity was more abundant in transformed cells, despite their lower actin content. Their membranes were also more effective in nucleating G-actin polymerization, indicating the presence of a greater number of filament ends.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990069 TI - Varicose veins as a source of adult human endothelial cells. AB - Endothelial cells can be harvested from segments of adult human saphenous vein in a varicose condition removed from patients having single or bilateral vein ligation and stripping. The cells are harvested by scraping with a scalpel, seeded on to gelatin coated or Primaria flasks and are passaged by removal with a rubber policeman. The cells cultured in this manner are maintained in a growth medium that is not supplemented with growth factors. The cells grow with a cobblestone monolayer morphology, possess angiotensin converting enzyme activity and react with antibodies to Factor VIII antigen. The cells fluoresce brightly after reaction with monoclonal antibodies specific for human endothelial cells. Thus, stripped varicose vein segments provide a readily available source of endothelial cells. PMID- 2990070 TI - Allylamine cardiovascular toxicity: VI. Subcellular distribution in rat aortas. AB - The cardiovascular toxin allylamine (3-aminopropene) has been shown to concentrate in elastic and muscular tissues. In this study the 14C-moiety of [14C]allylamine was traced in aortas of adult Sprague-Dawley rats after intravenously injecting 30 microCi of [14C]allylamine (spec. act. = 0.4 mCi/mM). At 5, 10, 15 and 20 min after injection 33.3-29.8% of the 14C-moiety was sequestered in aortas; at 30 min 16.8% was still present. Subcellular fractionation of the postnuclear supernatant by isopycinic centrifugation in sucrose demonstrated that 5 min after administration of [14C]allylamine, the 14C moiety displayed a modal density peak of 1.20 g/ml. Similar activities were observed up to 30 min exposure. This modal density was similar to the distribution pattern of mitochondria based on analysis of malate dehydrogenase activities. As early as 20 min post-exposure, mitochondrial malate dehydrogenase activities of aortic mitochondria decreased, while cytosolic malate dehydrogenase activities increased, suggesting mitochondrial membrane perturbation. We suggest that the subcellular site for allylamine injury to the aorta is the mitochondrion. PMID- 2990071 TI - The effect of prenatal chlordane exposure on specific anti-influenza cell mediated immunity. AB - Previous studies in our laboratory have documented that in utero chlordane exposure caused a significant enhancement in the survival of the offspring to influenza A virus infection, and a depressed delayed type hypersensitivity (DTH) response to oxazolone. To correlate these 2 effects, we assayed influenza A virus specific DTH response, and found that it was significantly decreased in chlordane treated offspring. Virus-specific T-cell blastogenesis was also assayed in chlordane-treated animals. No significant differences due to the chlordane treatment were found in virus-specific T-cell blastogenesis, suggesting that the DTH depression did not result from a paucity of antigen-reactive T-cells. To determine whether enhanced survival was due, in part, to the effects of chlordane on virus replication, rather than on immunological alteration alone, the kinetics of influenza virus replication in the lungs of chlordane- and vehicle-treated animals were determined. In utero chlordane treatment caused no significant differences in in vivo virus replication. These data suggest that increased survival was due to a decrease in virus-specific DTH and its associated pathology. PMID- 2990073 TI - Measurements of reticuloendothelial system phagocytic activity in the rat after treatment with silica, liposomes, and cyclosporine. AB - Reticuloendothelial system phagocytosis was evaluated in vivo by the clearance and tissue distribution of 99mTc-albumin millimicrospheres in rats. Administration of i.p. silica 2 days prior to clearance measurement increased the half-life of millimicrospheres 24% compared with control animals (P less than 0.01). Impairment of phagocytic activity was maintained at 5 days to 22% over control values (P less than 0.01). Tissue distribution studies showed that liver uptake was reduced and splenic and pulmonary uptake was increased compared with control. Cyclosporine i.v. increased half-life by 14% 1 hr after administration )P = NS) and half-life returned to control values by 8 hr. Hepatosplenic shift was less marked than after silica treatment. Phospholipid liposomes i.v. produced prompt impairment of millimicrosphere clearance after 1 hr; half-life was prolonged 46% over control values (P less than 0.001 vs. control) and marked radiolabel shift to the spleen and lung was seen. Measurements done 18 hr after a dose of liposomes revealed an increase of 55% over control half-life (P less than 0.001). Liposomes appear to be potent, nontoxic--and, in some cases, reversible agents--for blockade of reticuloendothelial phagocytosis and examination of non phagocytic reticuloendothelial functions. These results help to explain some of the results obtained in pancreatic islet grafting and suggest that phospholipid liposomes may be useful in this form of transplantation. PMID- 2990072 TI - Clinical significance of cytomegalovirus viremia in bone marrow transplantation. AB - Cytomegalovirus (CMV) viremia was systematically studied in 56 patients having undergone bone marrow transplantation for leukemia or aplastic anemia. Of the patients who survived at least three months, 57% had CMV viremia with a frequency peak between the 7th and the 9th weeks. We describe possible clinical signs associated with viremia, particularly late peripheral and/or central thrombocytopenia. The occurrence of viremia was studied according to the specific preexisting immune status of recipients and donors; granulocyte transfusions and graft-versus-host disease. The relationship between these parameters and viremia provides a basis for the analysis of prophylactic treatments of CMV infection. PMID- 2990074 TI - [Genetic transformation of somatic cells. II. An analysis of the status of the plasmid nucleotide sequences in chromosomal DNA and the thymidine kinase activity in transformant clone cells]. AB - Chinese hamster A238 TK- -cells were transformed with plasmids (derivatives of pBR325) containing thymidine kinase (TK) gene of Herpes simplex virus type 1 (HSV1). The results of dot- and blot-hybridization indicate the presence of pBR325 sequences in the chromosomal fractions of DNA in the transformant clones. These sequences are probably tandemly arranged, and each cluster contains 25--50 copies. SV40 sequences cloned in pBR325 were introduced into the Chinese hamster cells by co-transformation with TK-gene of HSV1-containing plasmid DNA, and all the co-transformant clones selected for TK+-phenotype were shown by hydridization to contain 3V40 DNA fragments. Isoelectrofocusing in polyacrylamide gel shows that thymidine kinase from TK+-transformant clones is of viral type (isoelectric point 7), in contrast to the cellular enzyme (coded by chromosomal gene) having alkaline isoelectric point (pH 9). The results suggest that the true TK+ transformant cells are selected by the procedure used in this study. PMID- 2990075 TI - [Genetic transformation of somatic cells. III. An analysis of the status of the plasmid nucleotide sequences in the extrachromosomal DNA of transformant clone cells and the rescue of extrachromosomal molecules of the plasmid DNA]. AB - Extrachromosomal DNAs from TK+ transformant clones of A238 Chinese hamster cells isolated after the treatment with plasmid pST826 containing thymidine kinase gene (TK-gene) of Herpes simplex virus (HSV1) and 1.8 kb insert of human satellite III DNA (HSIII) were studied by hybridization technique. In two TK+-clones (2T301 and 2T16) large quantities of rearranged plasmid DNA molecules were found. Electron microscopy show in clone 2T301 the presence of circular DNAs with average length being 4.64 +/- 0.27 kb. These molecules were rescued by retransformation into E. coli and analysed by restriction mapping and hybridization. All of them contain deletions spanning the entire TK gene of HSV1 and pBR325 sequences situated just downstream from the ORI of replication. The origin of extra-replicating circular DNA in 2T301 clone is discussed. PMID- 2990076 TI - [Genetic transformation of somatic cells. IV. The rate of loss of transformant phenotype depends on the structure of the transforming DNA and changes when the cells are treated with the tumor promotor 12-o-tetradecanoyl-phorbol-13-acetate]. AB - Analysis was made of the phenotype stability of some clones of thymidine kinase deficient (TK-) Chinese hamster cells transformed by thymidine kinase gene (TK gene) of Herpes simplex virus type (HSV 1). The presence of a fragment of human satellite DNA III in the plasmid DNA carrying the TK-gene of HSV 1 reduced notably the rate of the loss of TK+-phenotype, and the treatment of the cells with a tumour promoter--12-o-tetradecanoyl-phorbol-13-acetate--immediately after transformation destabilizes TK+-phenotype of transformant clones. Removal of the eukaryotic carrier DNA for the plasmid DNA without the TK-gene of HSV 1 destabilizes the clone transformant phenotype. Changes in the structure of the plasmid DNA containing no TK-gene of HSV 1 and introduced into cells simultaneously with TK-gene containing plasmids affects the rate of the loss of TK+-phenotype transformed cells. PMID- 2990077 TI - Localization, incidence, diagnosis and treatment of extratesticular germ cell tumors. AB - On the basis of a literature search yielding 243 case histories, the rare extratesticular germ cell tumor is surveyed with respect to incidence, primary localization, histology and treatment. In this context, we report on 6 patients with negative results on clinical examination of the testes (palpation, scrotal ultrasonography). We briefly discuss a suitable method of diagnosing occult and burnt-out tumors. Polychemotherapy (vinblastine, bleomycin, cis-platinum) has been found to be very effective. The resulting high rates of remission thus lead to an improvement of overall prognosis. PMID- 2990078 TI - Marijuana use does not spuriously elevate serum human chorionic gonadotropin levels. AB - Marijuana use has been reported to spuriously elevate immunoreactive human chorionic gonadotropin (HCG) in the serum of patients with testicular germ cell tumors. To reinvestigate this finding, we measured serum HCG and delta 9 tetrahydrocannabinol (delta 9-THC) levels in 16 men known or suspected to be marijuana users. Eight of the serum samples had measurable levels of delta 9-THC, while eight did not. None contained immunoreactive HCG. The addition of delta 9 THC directly to pooled male serum had no effect on the HCG radioimmunoassay. These results indicate that marijuana does not artificially elevate serum HCG concentrations. PMID- 2990079 TI - Kinetics of formation of renotropic growth factor. AB - In the Wistar/Furth rat, twenty-four hours after unilateral nephrectomy, the remaining contralateral kidney produces maximum amounts of renotropic growth factors. These factors stimulate tumor growth in recipient rats with s.c. Wilms tumor, but have no effect on kidney weight. In tumor-free recipients, kidney hypotrophy results when the factors are administered. PMID- 2990080 TI - Pattern of calcium metabolism in normo- and hypercalciuric patients with calcium urolithiasis in Saudi Arabia. AB - Urinary calcium excretion was measured in 100 consecutive normocalcemic patients with calcium urolithiasis and 12 Saudi Arabian controls while the patients and the controls were eating their usual diet of unknown calcium content. Only 16 patients were hypercalciuric using the definition of twenty-four-hour urinary calcium of more than 300 mg for males and more than 250 mg for females, and one of these patients was subsequently found to have hyperparathyroidism. The twenty four-hour urine calcium was less than 200 mg in 60 per cent of male patients and in all the male controls. Since the dietary intake of calcium during the twenty four-hour urinary collection was unknown, a 1,000-mg calcium loading test was performed in an attempt to differentiate various patterns of abnormal calcium excretion. Of the twenty-four-hour normocalciuric patients 10 (18%) had "absorptive hypercalciuria" and 16 patients (29%) demonstrated a "renal hypercalciuria" pattern. Thirty-nine patients (57%) and all 12 controls had normocalciuria before and after calcium loading. PMID- 2990082 TI - Experimental infection of a natural case of sheep pulmonary adenomatosis with maedi-visna virus. PMID- 2990081 TI - Malignant fibrous histiocytoma of prostate gland. AB - The English language medical literature does not contain any definitive reports of malignant fibrous histiocytoma (MFH) arising in the prostate gland. A case of prostatic MFH is described, characterized by rapid local tumor progression and pulmonary metastases. The histologic diagnosis was supported by immunohistochemical and electron microscopic studies. PMID- 2990083 TI - Culicoides imicola: a bluetongue virus vector in Spain and Portugal. PMID- 2990084 TI - Parturition as a stimulus of IBR virus reactivation. PMID- 2990085 TI - Use of by-products and supplementary protein in dairy cow nutrition. AB - The new systems of protein evaluation distinguish clearly between the requirement of the rumen microbes and the need of the host animal. While the requirement of the rumen microbes and the production of microbial protein is related to fermented energy, the need of the host animal varies with type and level of productivity. In dairy cows microbial protein is insufficient for the host animal, particularly when the animals are in negative energy balance, and in order to efficiently utilise body energy reserves undegraded protein must be given. Several experiments are discussed in which dairy cows have been given ammonia treated straw as the only source of roughage and where milk yields have been maintained. It is emphasised that increased use of high roughage diets in early lactation increases the demand for undegraded proteins. The converse is also true: that undegraded protein increases the proportion of roughage that can be used for high yielding dairy cows. PMID- 2990086 TI - Disease in turkeys associated with H1N1 influenza virus following an outbreak of the disease in pigs. PMID- 2990087 TI - A chronic indurative mastitis in sheep, associated with maedi/visna virus infection. AB - A possible association of a chronic indurative mastitis with mvv (maedi visna virus) infection in sheep was investigated. Sheep of four flocks (group A), in which insufficient lamb growth apparently associated with insufficient milk production and probably related to udder induration, was a serious problem, were clinically examined in mid-lactation. The results were compared with four mvv free flocks (group B) without such complaints. The incidence of udder induration in group A (n = 263) was about eight times higher than in group B (n = 206): 63.1 versus 8.0%. The clinical picture differed essentially between the groups. In group A the udder abnormalities were of a diffuse and indurative nature, involving both udder halves, while in group B the udder lesions were mainly nodular and often limited to one udder half. Bacteriological examination revealed a difference in infection rate of the udders (6.8% in group A versus 14.1% in group B). A substantial difference was observed in a comparison of the bacteriological infection rate of the clinically abnormal udders (5.4% in group A, versus 47.0% in group B). Serological examination with an indirect ELISA revealed 81% seropositive sheep in group A, versus 0% in group B. Twelve sheep of group A and five of group B, called for reason of mastitis, were selected for pathological examination. The gross and microscopic lesions showed a pronounced difference between both groups. In group A a diffuse interstitial mastitis with slight to moderate fibrosis and a pronounced lymphoid hyperplasia was observed. In group B the mastitis had a nodular character, with a chronic galactophoritis, extensive fibrosis, and in some cases chronic abscesses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990088 TI - A case of chronic zinc poisoning in calves fed with zinc-contaminated roughage. AB - A case of zinc intoxication in young female cattle is described. The clinical signs consisted of reduced appetite, emaciation, submandibular oedema and diarrhoea. The source of zinc proved to be roughage harvested in the vicinity of a factory galvanizing steel tubes. In this roughage zinc levels between 3000 and 7300 mg/kg dry weight were found. In the liver of four animals zinc levels varied between 420 and 1600 mg/kg, and between 910 and 1680 mg/kg dry weight in the kidneys. PMID- 2990089 TI - The use in practice of inactivated oil emulsion vaccine against infectious bursal disease in broiler breeders and its influence on the progeny: a comparative field trial. AB - Field trials were conducted to establish the effect of the use of an inactivated oil emulsion vaccine against Infectious Bursal Disease (IBD OEV) in broiler breeder hens, and its effects on their progeny. The performance of 18 broiler flocks, which were the progeny of the IBD OEV vaccinated breeder hens, but which were not vaccinated with a live vaccine against IBD, was equal to that of broiler flocks which were vaccinated with a live IBD vaccine and originated from parent stock that had been vaccinated only against IBD with a live vaccine. In none of the 18 flocks, progeny of IBD OEV vaccinated parents, was IBD diagnosed. In a second stage, 15 broiler flocks were included in the trial: these were derived partly from IBD OEV vaccinated parents, and partly from parents that received only live IBD vaccine at 8-10 days of age. No cases of IBD occurred and all flocks were positive for IBD precipitins at slaughter age. Vaccination with a live vaccine against IBD at the age of 8-10 days had no influence on NCD antibody development after a NCD vaccination at 7 days. No immunosuppressive effect from this type of live live IBD vaccine could be determined under field conditions. PMID- 2990090 TI - [Value of lymphography in the diagnosis and treatment of rectal cancer]. PMID- 2990091 TI - [Role of mammography in the preoperative diagnosis of fibroadenomas of differing histological structure]. PMID- 2990092 TI - [Experiments to produce an attenuated strain of the transmissible gastroenteritis virus and its use as a live vaccine]. AB - Serial passages have been performed of a field virus of the transmissive gastroenteritis to obtain an attenuated strain adapted to a permanent cell line of pig kidney, "SPEV". The strain is innocuous for test animals, for pigs of all ages, and for swine during the entire period of pregnancy. It has proved genetically stable. The strain is immunogenic for swine and pigs both under laboratory and under field conditions (in new foci and on stationary farms) and can be used as a live vaccine against transmissive gastroenteritis, either untreated or in a freeze-dried state. PMID- 2990094 TI - [Isolation of the transmissible gastroenteritis virus in a continuous pig embryo kidney cell line]. AB - Attempts were made to isolate the virus of transmissive gastroenteritis in a permanent cell line SPEV in view of the diagnostics of the disease. Used were small intestines of pigs during the first 24 hours after the setting in of clinical symptoms. The successful isolation of the virus in a SPEV line depended on the methods employed to handle the material as well as on the ways of inoculating the cell cultures. In the conditions of the investigation most proper proved centrifugation of the organ suspensions and the suspension method of infection. The SPEV line, however, was shown to be insufficiently sensitive to isolate the virus of transmissive gastroenteritis. About 30 per cent only of the positive material could be used to demonstrate the virus following direct infection, while the study of the remaining (up to 62 per cent) material required additional passing. The SPEV line could replicate the virus after its infection with organ suspensions containing the virus, and this could be demonstrated through immunofluorescence investigations. The same method could likewise be employed in the express diagnosing of the disease. The passing of the virus strains in the SPEV line led to their attenuation. In order to retain the virulence of the strains passing should take place in nonimmune pigs or should alternate with the use of both pigs and cell cultures. PMID- 2990093 TI - [Bactericidal effect of the preparation trozin]. AB - Tested was the bactericidal action of the combined cleansing and disinfecting preparation trozin under laboratory and production conditions. It was found that trozin meets the requirements in terms of rapid disinfection, killing the asporogenous test organisms. The presence of at least 2 per cent milk protein lowers the bactericidal action of the detergent. The cleansing and disinfection of milking units supplied with a milk-can, resp., with a central milk pipeline leads to 97.00, resp., 98.00 per cent reduction, on an average, of microflora. The bacterial count of the udder is likewise reduced by about 96.00 per cent. PMID- 2990095 TI - The Epstein-Barr virus nuclear antigen (BamHI K antigen) is a single-stranded DNA binding phosphoprotein. AB - The Epstein-Barr virus BamHI K nuclear antigen was shown to be phosphorylated in latently infected and virus-producing B-cell lines by in vivo labeling of cell cultures with [32P]orthophosphate and immunoprecipitation with anti-BamHI K antigen monoclonal antibody. Phosphoamino acid analysis of this protein isolated from a latently infected cell line demonstrated that the modified amino acid is phosphoserine. The BamHI K nuclear antigen transiently expressed in NIH 3T3 cells is also phosphorylated, as well as three truncated and deleted forms of the protein. Interaction of the Epstein-Barr virus BamHI K nuclear antigen with denatured DNA was examined by chromatography of wild-type and mutant forms of this protein on single-stranded DNA cellulose columns. The wild-type protein bound to denatured DNA cellulose but not cellulose alone. The BamHI K antigen remained bound to single-stranded DNA in 300 mM NaCl and eluted from the DNA at higher NaCl concentration. Similar results were obtained with 32P-labeled protein and total antigen as assayed by radioimmunoelectrophoresis. A mutant protein that lacks the glycine and alanine repeated amino acid domain and surrounding amino acids of this EBV polypeptide retained the ability to bind to denatured DNA, although it eluted at slightly lower NaCl concentration. One mutant protein that lacks the carboxyl-terminal third of the protein failed to bind to single stranded DNA. PMID- 2990097 TI - Avian retrovirus S13: properties of the genome and of the transformation-specific protein. AB - The avian retrovirus S13 codes for an env-linked transformation-specific glycoprotein with a molecular weight of 155,000 (gp155). Treatment of gp155 with endoglycosidase H or growth of S13-infected cells in the presence of tunicamycin reduces the molecular weight of gp155 to about 140K, but these gp155-related molecules may still contain sugar residues. The gp155 protein is not incorporated into virions; it is phosphorylated, but in immunoprecipitates does not show protein kinase activity. The genome of S13 is an 8.5-kilobase (kb) RNA; the helper virus genome is 7.5 kb in size. The putative onc sequences of S13 do not hybridize to DNA probes representing src, erb A, erb B, myc, myb, fps, fms, H ras, B-lym, abl, rel, and ets. PMID- 2990096 TI - S13, a rapidly oncogenic replication-defective avian retrovirus. AB - The avian leukemia sarcoma virus S13 transforms chicken and Japanese quail embryo fibroblasts and chicken erythroid cells in tissue culture. S13-induced erythroid transformation requires culture conditions suitable for the growth of normal erythroid precursors (H. Beug and M. J. Hayman (1984), Cell 36, 963-972). S13 transformed erythroid colonies contain a high percentage of cells that differentiate in absence of erythropoietin. S13 is defective in pol and env functions but can code for a complete set of gag proteins. Nonproducer cell clones transformed by S13 release a noninfectious viral particle containing gag but no functional env or pol proteins. They also synthesize a transformation specific protein of 155,000 molecular weight. This protein reacts with antibody to viral envelope glycoproteins and appears to represent onc as well as env sequences. The 155,000-molecular weight env-linked protein does not cross react immunologically with an antiserum against the v-erb A and v-erb B gene products. PMID- 2990098 TI - Cell-specific selection of mutants of a herpes simplex virus recombinant carrying deletions. AB - Herpes simplex virus 1 (HSV-1) recombinant R316 was constructed so as to convert the thymidine kinase (TK), a beta gene, into an alpha-regulated gene by insertion of the BamHI N fragment in the proper transcriptional orientation into the BglII cleavage site of the TK gene (L. E. Post, S. Mackem, and B. Roizman, Cell 24, 555 565 (1981).) The BamHI N fragment contains the promoter and regulatory domains of the alpha 4 gene in addition to an origin of viral DNA synthesis and the complete domain of the alpha 22 gene. Passage of the R316 virus in HEp-2 or in human embryonic lung (HEL) cells resulted in rapid accumulation of mutants carrying approximately 4.4-kbp deletions in the insert. No appreciable accumulation of the deletions was observed upon passage of R316 virus in Vero cells. The accumulation of deletions in HEp-2 and HEL cells could not be attributed to the fusion of the TK gene with the alpha 4 gene promoter, to the presence of an origin of DNA replication, or to overexpression of any of the genes whose domain is contained entirely in the HSV-1 Bam HI N fragment; these conclusions are based on the observations that deletions did not accumulate in HEL or HEp-2 cells infected with recombinant R315, containing BamHI N inserted in an inverted orientation, or with recombinant R314, carrying an alpha 4-TK chimera constructed by insertion of the HSV-1 BamHI Z fragment into the BglII cleavage site in the TK gene. The BamHI Z fragment also contains a functional origin of DNA synthesis. The hypothetical models which could explain the host range-specific accumulation of deletions are discussed. PMID- 2990100 TI - Establishment of cell lines persistently infected with foot-and-mouth disease virus. AB - Cell lines persistently infected with foot-and-mouth disease virus (FMDV) have been established by growth of BHK-21 (c-13) or IBRS-2 (c-26) that survived standard cytolytic infections with FMDV. They maintain cytoplasmic FMDV RNA sequences, as shown by dot blot hybridization tests, using cloned FMDV cDNA as probes. Cell line C1-BHK-Rc1 was derived by infection of cloned BHK-21 c1 cells and plaque-purified FMDV C-S8 c1. Indirect immunofluorescence assays indicated the presence of FMDV antigens. It was resistant to superinfection by FMDV C-S8 c1, O-S7, or A5, but not by encephalomyocarditis virus (EMCV), vesicular stomatitis virus (VSV), or Semliki forest virus (SFV). Infectious FMDV was detected in the culture medium only up to cell passage 65. The virus isolated from C1-BHK-Rc1 cells showed decreased plaque size and diminished yield in infections at 42 degrees. Multiple mutations in the intracellular FMDV RNA have been detected by T1 oligonucleotide fingerprinting of genomic RNA segments hybridized to FMDV cDNA fragments. At late cell passages, when no infectious FMDV is detected, cells continue to express viral antigens and FMDV RNAs with deletions of up to 3 kb have been identified by Northern blot analysis. We conclude that persistent infections of cell cultures with FMDV are readily established and that multiple genetic and phenotypic variations occur in the virus during persistence. PMID- 2990101 TI - Nucleotide sequence of a herpes simplex virus type 1 gene that causes cell fusion. AB - The nucleotide sequence (2041 nucleotides) of a genomic region of herpes simplex virus type 1 (KOS strain) associated with virus-induced cell fusion has been determined. The sequence is bounded by a NruI site at 0.732 and a BamHI site at 0.745 prototypic map units. An open reading frame in the left-to-right orientation specifies a protein of 338 amino acids. The protein is positively charged. Since secondary structure analysis predicts four extensive hydrophobic domains the protein is probably a membrane-associated or a transmembrane protein. Transcription of the putative fusion gene is dependent on viral DNA synthesis, characteristic of the late (gamma) viral gene class. Two syncytia-inducing mutations, syn20 and MP, have been previously mapped to a 504-base pair PstI fragment within these genomic coordinates (V. C. Bond and S. Person (1984), Virology 132, 368-376). The nucleotide sequence of the PstI fragment was determined for the two mutants. Both were shown to have an amino acid substitution at residue 40 of the fusion protein. A second change at residue 101 for MP is probably unrelated to the fusion phenotype. PMID- 2990099 TI - Human papillomavirus type 16 DNA sequence. AB - The complete nucleotide sequence of HPV16 DNA (7904 bp) cloned from an invasive cervical carcinoma was determined. Homology comparisons allowed us to align the major open reading frames with the other published papilloma virus DNA sequences. The general organization of the open reading frames is similar to that of the other four papillomavirus (BPV1, HPV1a, HPV6b, CRPV) already sequenced. The sequence reveals an interruption of the reading frame coding for a suspected E1 protein. PMID- 2990102 TI - Equine herpesvirus type 1 infected cell polypeptides: evidence for immediate early/early/late regulation of viral gene expression. AB - EHV-1 polypeptide synthesis was examined in productively infected rabbit kidney and hamster embryo cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses of extracts from [35S]methionine- and 3H amino acid-labeled-infected and mock-infected cultures revealed the presence of 30 infected cell-specific polypeptides (ICPs) which ranged in apparent molecular weights from 16.5K to 213K. Twenty-two of these ICPs comigrated with virion structural proteins. Four ICPs (203K, 176K, 151K, 129K) were detected in extracts of infected cultures labeled in the presence or absence of actinomycin D (Act D) immediately after release from a 4-hr treatment with cycloheximide (CH). These polypeptides, which were designated as EHV-1 immediate early (alpha) ICPs, were not detected in unblocked (non-CH-treated) infected cells. The most abundant ICP was a 31.5K nonstructural protein which, in addition to a 74K protein, was detected in unblocked infected cells at 2-3 hr postinfection. These proteins appeared to be regulated as early (beta) ICPs, since neither protein was observed in Act D-treated cultures released from CH block. Twelve ICPs were classified as late (gamma) polypeptides on the basis of their reduced synthesis in cultures in which viral DNA replication was inhibited by phosphonoacetic acid. All but one (40K) of these late ICPs corresponded to virion structural proteins. PMID- 2990103 TI - Varicella-zoster virus envelope glycoproteins: biochemical characterization and identification in clinical material. AB - Varicella-zoster virus (VZV)-infected human foreskin fibroblasts synthesize viral glycoproteins of 125,000 (gp125), 118,000 (gp118), 92,000 (gp92), 63,000 (gp63), 59,000 (gp59), and 47,000 (gp47) Da. In biochemical studies, all of these VZV glycoproteins were shown to contain asparagine-linked (N-linked) oligosaccharide chains and, except for gp125 and gp47, to be sialoglycoproteins. Experiments with endo-beta-N-acetylglucosaminidase H (endo H) demonstrated that gp92 contained only complex type (endo H-resistant) N-linked glycosyl chains, while the other mature glycoproteins contained both high-mannose (endo H-sensitive) and complex type oligosaccharides. Monoclonal antibodies recognizing multiple glycoproteins, gp63/gp125 or gp92/gp59/gp47, neutralized virus infection, suggesting the glycoproteins were important components of the virus envelope. This was confirmed for gp92/gp59/gp47 by immunoelectron microscopy, which revealed dense staining localized exclusively to the virion envelope and to the plasma membrane of virus producer cells. The mature forms of all of these glycoproteins were also present in viral material isolated from vesicles of varicella and zoster patients, indicating that in infected individuals the viral glycoproteins are synthesized and processed in a manner similar to that in tissue culture cells. PMID- 2990104 TI - Inhibition of HSV-transformed murine cells by nucleoside analogs, 2'-NDG and 2' nor-cGMP: mechanisms of inhibition and reversal by exogenous nucleosides. AB - A murine cell line transformed with HSV TK (LH-1) exhibits a greatly enhanced cytotoxicity to the nucleoside analog 9-[(2-hydroxy-1-(hydroxymethyl)ethoxy) methyl]guanine (2'-NDG) as compared to the parental LM cell line (I50 LH-1 = 0.4 microM; I50 LM = 44.4 microM). Toxicity of 2'-NDG for LH-1 and LM is reversed only by the addition of 100 microM thymidine (dThd), indicating that 2'-NDG is a substrate for the viral and cellular TK. In LM(TK-) cells--murine cells expressing no TK activity, 2'-NDG cytotoxicity is partially reversed only with dGuo. A cyclic phosphate derivative of 2'-NDG, 2'-nor-cGMP, contains a phosphodiester bond, is also taken up by cells, and does not depend on viral TK for activation. LH-1 cells and LM(TK-) cells are inhibited by similar concentrations of this analog (5.1 and 4.1 microM, respectively). In all three cell lines (LM, LH-1, LM(TK-], the toxicity of 2'-nor-cGMP is significantly reversed with dGuo or cyclic dGMP. This pattern of reversal differs significantly from that observed with 2'-NDG, suggesting that 2'-nor-cGMP is metabolized as a guanosine analog, similar to acyclovir, in LM and LM(TK-) cells. These results indicate that a cyclic monophosphate analog of 2'-NDG can be activated independently of viral TK expression and that cellular metabolic pathways resulting in elevated dGTP concentrations are important for reversal of toxicity induced by guanosine-like nucleoside analogs. PMID- 2990105 TI - [Blood hormone levels in persons of various blood groups]. PMID- 2990106 TI - [Tubular carcinoma of the breast (clinico-morphological characteristics)]. AB - The following four types of tubular carcinoma of the breast were identified as a result of the study of 39 cases: typical tubular carcinoma of the breast; tubular mammary carcinoma with ductal invasion; ductal invasive carcinoma with tubular structures predominating over more than half the area of section; ductal invasive carcinoma incorporating few tubular structures. A specific clinico-morphological picture and prognosis are peculiar to each type of carcinoma. Tubular carcinoma of the breast and ductal invasive tumor with predominant tubular structures have a favorable prognosis and a regional metastasis frequency of 23-28%, practically without distant metastases. Tubular carcinoma of the breast is considered an early form of ductal invasive carcinoma which develops into other histological patterns as tumor growth progresses. PMID- 2990107 TI - [Biochemical effects of the continuous impulse electromagnetic field (lambda-35 cm)]. PMID- 2990108 TI - [Endorphins, enkephalins and their antagonists in clinical practice (review of the literature)]. PMID- 2990109 TI - Intraperitoneal rupture of a Wilms' tumor. PMID- 2990110 TI - [Nephroblastoma with metastasis to the heart simulating portal hypertension syndrome in an 8-year-old boy]. PMID- 2990112 TI - [Diagnosis, therapy and prognosis of tumors of the skin in the aged]. PMID- 2990111 TI - [Jugulotympanic paraganglioma]. AB - A case is described of a jugulotympanic paraganglioma in a 64-year-old woman with extentions down the jugular vein to the clavicle and penetration of the cerebellar fossa. The symptoms, differential diagnosis, therapy and prognosis are discussed on the basis of this case report and compared with the literature. The importance of radiological investigation (e.g. selective angiography and computed tomography) in respect to diagnosis and determination of tumour size is emphasized. The diagnostic value of the immunohistochemical detection of neuron specific enolase, a neuroendocrine cell marker and S-100 protein is stressed. The most favourable therapy, depending on tumour extension, seems to comprise preoperative embolisation, radical resection and postoperative radiotherapy. PMID- 2990113 TI - [Relevance of serum aldolase for the detection of liver metastases of bronchial cancer--comparative enzymatic and endoscopy-autopsy studies]. AB - In 100 patients with histologically confirmed bronchogenic carcinoma, the reliability of serum aldolase determination as screening test for metastases in the liver has been investigated. Demonstration respectively exclusion of liver metastases was done by laparoscopy or occasionally by autopsy. In contrast to previous results, with the presently used, more reliable technique, the results were less satisfactory. True positive aldolase values as indicators of presence of liver metastases were observed in 72,7 percent of patients carrying those. In cases of chronic liver damage of other etiology, in general no elevated aldolase values were observed. The aldolase activity correlated with the amount of tumor tissue in the liver. An improvement of the enzymatic screening might be expected from additional determination of enzymes indicating cholostasis or from determination of liver-specific aldolase isoenzyme. PMID- 2990114 TI - [Respiratory function disorders caused by agrochemical exposure]. AB - With the aim to evaluate possible influences of exposition to agrochemicals upon the respiratory system, the ventilatory parameters VC, FEV1,0, FIV1,0, and the values of PaO2 and PaCO2 were investigated in 93 males professionally exposed to mineral fertilizers, and in 69 males professionally exposed to pesticides--both groups working in agrochemical centres. A control group consisted of 162 males without exposition to agrochemicals. In exposed workers mean values of VC, FEV1,0, and PaO2 were lower (highly significant) compared with the control group. Mean values of FEV1,0, FIV1,0, and PaO2 in workers exposed to mineral fertilizers were in the borderline region. Influences upon the respiratory system should be further investigated with regard to long-term effects and to fitness for the job. PMID- 2990116 TI - [Survival time of enterobacteria and enteroviruses in activated sludge-fertilized soils]. PMID- 2990115 TI - [The future of viral hepatitis vaccines]. PMID- 2990117 TI - [Paraneoplastic multicentric reticulohistiocytosis: induced and inductable by iodine containing x-ray contrast media]. AB - Following lymphography, a 41-year-old woman developed arthritis and papules of multicentric reticulohistiocytosis three years after the beginning of a carcinoma of the cervix, now inoperable and with vulvar metastases. Two further inevitable x-ray investigations with different contrast media led to exacerbation of the disease within 24 hours. 12 month after onset, all symptoms of multicentric reticulohistiocytosis receded spontaneously, and further investigations with contrast media were well tolerated. We assume that iodine compounds may be the cause--directly or indirectly--for provocation and enhancement of histiocytic proliferation. PMID- 2990118 TI - [Metabolic effect of neurohypophyseal hormones]. PMID- 2990119 TI - [Cellular and molecular basis of the participation of prostaglandins in the function of the nervous system]. PMID- 2990120 TI - [Current concepts of the principles of odor discrimination in the olfactory organ of vertebrates]. PMID- 2990122 TI - Maternal and fetal effects of epidural analgesia during labour. AB - We can state that an effective pain relief during labour, like epidural analgesia, is indicated and it prevents or modifies the pain-induced harmful effects on mother and the fetus. Maternal and fetal ACTH or BE response in labour are not altered, but the increase of maternal cortisol secretion is reduced, thus probably suggesting a lesser maternal stress, to which also point the changes of PRL. Epidural analgesia prevents harmful maternal metabolic changes like hyperventilation, hypocapnia, metabolic acidosis and lactic acid accumulation. These beneficial effects are also reflected in a better fetal and neonatal condition. Carbohydrate and fat metabolism, on the contrary, seem not to be significantly changed by epidural analgesia. Epidural analgesia is beneficial also by reducing the pain induced catecholamine release, and by improving the intervillous blood flow, especially in pre-eclamptic parturients. PMID- 2990121 TI - [Papilloma virus-induced changes in the uterine cervix. Histo- and cytologic and epidemiologic findings]. AB - In this epidemiological, cytological and histological study signs of papillomavirus infections were found in 128 cases (16,6 per cent) and dysplasia or carcinoma in situ in 713 cases in a series of 773 cone biopsies. Epithelial changes caused by papillomavirus were combined with preneoplastic lesions in 116 cases. Pretherapeutic cytologic examinations of the condylomatous epithelial changes secured histologically later on referred to cervical papillomavirus infections in 64 per cent. Epidemiological inquiries of patients with cervical papillomavirus infection correspond extensively to those with invasive carcinoma of the cervix and premalignant cervical lesions. PMID- 2990123 TI - [Development of a numerically additive combined vaccine against tetanus and smallpox]. AB - Mandatory vaccination against smallpox was abolished on the account of smallpox eradication proclaimed by the WHO and the postvaccinal complications detected after smallpox vaccination. At the same time vaccine banks with the vaccinia virus strain "Elstree" were organized. Should mass vaccinations with this vaccinia virus strain be carried out in a case of emergency, severe postvaccinal diseases and complications can arise in overaged and immunosuppressed vaccinees after primovaccination. Therefore attenuated vaccinia virus strains should be used for vaccine banks, which cannot be activated, or increase in virulence in impaired vaccinees after primovaccination. For these individuals the vaccinia virus strain "MVA", among other attenuated vaccinia strains, is recommended. The MVA virus strain can be applied parenterally without complications. From the scientific and field-relevant point of view it was tried to combine the vaccinia virus strain "MVA" with tetanus toxoid and to develop a combination vaccine "tetanus-smallpox". In immunization experiments using mice, piglets and monkeys, safety and efficacy of the vaccine were investigated. Efficacy was demonstrated by means of postvaccinal antibody determination and by the mouse protection test. Tetanus antitoxin was measured by ELISA and indirect hemagglutination test, antibody levels to vaccinia virus were investigated employing the neutralization test and hemagglutination inhibition test. No significant differences in potency could be demonstrated between the combination vaccine and the corresponding monovalent vaccines in mice, piglets and monkeys. The combination vaccine consisted of 12 Lf tetanus toxoid and 10 TCID50 vaccinia virus "MVA" preserved with gelatine and glucosamine. The double intramuscular immunization of monkeys stimulated average tetanus antitoxin titers of 1:310 and average vaccinia virus titers of 1:195 2 weeks p. revacc. Similar results were obtained in mice and piglets. Side reactions were not observed in mice and piglets. Except for occasional local reactions of short duration at the injection site of the monkeys, similarly no adverse reactions were observed after intramuscular vaccination with the combination vaccine. PMID- 2990124 TI - Incidences of antibodies to hepatitis B, herpes simplex and cytomegalovirus in prostitutes. AB - The 170 prostitutes registered by the local public health authority (165 female, 5 male; ages 17-61) were serologically investigated by ELISA and CFT for the presence of hepatitis B virus (HBV) markers (n = 170), anti-CMV (n = 118) and anti-HSV 2 antibodies (n = 117). 4.7% (1.2%) were found to be HBs- (HBe-)antigen carriers; 1.2% revealed an acute hepatitis B (seroconversion of anti-HBc or IgM anti-HBc highly positive); 17.1% presented a past and resolved hepatitis B (anti HBs positive). These rates exceeded those of 125 female age-matched control persons considerably, out of whom only 2.4% (0.8%) presented HBs-(HBe-) antigen and 3.2% anti-HBs antibodies. 90% of the prostitutes investigated proved to be anti-CMV IgG (1.7% IgM) antibody positive and were compared to healthy blood donors (47% IgG, 0% IgM-anti-CMV antibody carriers) and other CMV risk groups: pregnant women 56 (13), patients in hemodialysis 61 (0), hemophiliac patients 69 (0), patients after kidney transplantation 90 (16) and after open-heart surgery 87 (4) % anti-CMV IgG (IgM) antibody carriers. An AIDS patient suffered from simultaneous HBV and CMV infections. The risk of HSV 2 infection is among prostitutes (38.5% seropositives) five times as high as among female age-matched control persons (7.2% seropositives, n = 125). These results confirm that prostitutes must be regarded as a risk for the spread of hepatitis BV and CMV- and HSV 2-caused diseases in the population. PMID- 2990125 TI - [Search for the systems of DNA host specificity in Salmonella typhi]. AB - Seventeen pure lines of S. typhi bacteriophages have been obtained from mother races O and Vi; of these, three were used to study 152 S. typhi strains with a view of detecting their DNA host specificity systems. 10 S. typhi strains having the DNA host specificity system have been detected by the rough determination of the lytic spectrum and the cross titration of phage Vi IX. PMID- 2990126 TI - [Standardization of the conditions for measuring the specific fluorescence intensity in continuous cell cultures infected with variants of the Japanese encephalitis virus]. AB - The use of modernized photometric accessories to a model microscope manufactured by the Leningrad Optico-Mechanical Amalgamation (USSR), as well as the practical application of innovations facilitating and standardizing research work, has made it possible to obtain objective data indicating the presence of significant, direct, linear correlation between infectious activity and the intensity of fluorescence emitted by fluorescent antibodies bound with the antigen of 11 studied variants of Japanese encephalitis virus in continuous cell lines. The study of the dynamics of fluorescence intensity permitted the objective evaluation of the previously revealed regularity in the increase of the intensity of the induced fluorescence of Japanese encephalitis antigen in continuous cell cultures. PMID- 2990127 TI - [Current problems of the epidemiological surveillance of poliomyelitis]. AB - A uniform program of the epidemiological supervision of poliomyelitis must be worked out for every administrative territorial unit. This program should include the early and complete detection of all cases of this infection, irrespective of their severity; the analysis of the quality of vaccination; the determination of the immune status of the child population; the characteristic of the poliovirus circulation. PMID- 2990128 TI - [Phyllodes tumors of the breast]. AB - Clinical pattern compared with ultrasound, radiology and fine needle acubiopsy allows an exact diagnosis of cystosarcoma phyllodes before surgical operation. Then, even using quite conservative procedures on low-grade tumours, we had very good results. PMID- 2990129 TI - [Examination using osteotropic radionuclides in patients with advanced coxarthrosis]. PMID- 2990130 TI - Studies on the site of origin and molecular form of the extrahypothalamic corticotrophin-releasing factor (CRF) in rats. AB - A homologous radioimmunoassay was developed for rat CRF. CRF immunoreactivity was found widely in the extrahypothalamic areas in the rat central nervous system in concentrations 1-3% of that in the median eminence. Large hypothalamic lesions including the paraventricular nucleus and the median eminence did not cause a decrease in the extrahypothalamic CRF concentrations. CRF of the rat hypothalamus and cerebral cortex both consisted of high and low molecular weight forms. In contrast the posterior pituitary and the spinal cord contained only low molecular weight forms. Upon gel filtration the low molecular weight CRF of the hypothalamus co-eluted with synthetic rat CRF whereas that of the spinal cord was considerably more retarded. The low molecular weight CRF of the posterior pituitary and cerebral cortex was slightly more retarded compared to synthetic rat CRF. We conclude that CRF in many extrahypothalamic brain areas and the spinal cord does not originate from the hypothalamus and that there is some heterogeneity in the molecular form of the extrahypothalamic CRF. PMID- 2990131 TI - Nocturnal high-dose dexamethasone suppression test in the aetiological diagnosis of Cushing's syndrome. AB - Seventeen patients with well-proven Cushing's syndrome (13 with Cushing's disease, 3 with adrenal tumour and 1 presenting ectopic ACTH syndrome caused by bronchial carcinoid) were investigated by using a single-dose 8 mg dexamethasone nocturnal test. The results obtained were compared with those of the classical 8 mg Liddle's test, metyrapone stimulation, plasma ACTH concentration and with the final diagnosis reached through surgery, pathologic anatomy, and/or clinical and biochemical follow-up of the patients after treatment. The diagnostic efficacy or the predictive power of the test (defined as the ratio between the number of cases in which the diagnosis was correctly predicted and the total number of cases), was at least 82.4% vs 84.6% for the classical 8 mg Liddle's test. This percentage increased to 94.1% when the results of repeated tests on three patients with conflicting data were included. It is concluded that the nocturnal high-dose dexamethasone suppression test is a valuable tool in the aetiological diagnosis of Cushing's syndrome. PMID- 2990132 TI - Sex-dependent endorphinergic and adrenergic control mechanisms of luteinizing hormone secretion in immature rats. AB - Previous studies in male and female immature rats have revealed striking sex differences as concerns endorphinergic and adrenergic control of luteinizing hormone (LH) secretion. The present study examines in 10 days old male and females rats whether these differences result from sexual differentiation of the brain, or acute effects of male and female gonadal hormones. The techniques employed to manipulate these mechanisms were gonadectomy immediately post-partum and androgenization. Androgenization on the 1st and 2nd day of life reduces the ability of naloxone to elevate serum LH levels in females, but failed to modify the LH-elevating effect of clonidine in males. Experiments with castrates showed that testosterone is critical for these sex-related differences. Treatment with testosterone on the 9th day of life of intact or gonadectomized rats revealed the ability of this hormone to modify LH-release acutely. We conclude that sexual differentiation of the brain may be of minor significance for the sex-related LH control mechanisms in prepubertal rats. Of importance is the acute presence of testosterone, since in its absence male characteristics disappear. PMID- 2990134 TI - Sequential administration of human and porcine factor VIII for surgical treatment of a parotid tumour in a patient with a factor VIII inhibitor. AB - The sequential use of human and porcine factor VIII for the treatment of a patient with a moderately high titre (38 Bethesda units) factor VIII inhibitor enabled us to provide adequate haemostatic cover for 17 days. In this period a pleomorphic adenoma of the parotid gland was removed. Bleeding did not occur and wound healing was uneventful. PMID- 2990133 TI - Responses of patients with Bartter's syndrome to angiotensin III and angiotensin II-(3-8)-hexapeptide. AB - Studies were conducted to determine whether or not angiotensin III [AIII] and angiotensin II-(3-8)-hexapeptide [ANG-(3-8)] have their own specific arteriolar binding sites different from angiotensin II [AII] binding site(s) in man. Four patients with Bartter's syndrome were given asn1-,val5-AII by iv infusion at rates of 10, 20, 50 and 100 pmol/kg X min, each for 7 min. One hour later AIII was infused iv in the same 4 patients at rates of 50, 100, 250 and 500 pmol/kg X min, each for 7 min. After 100 or 150 mg/day of indomethacin treatment for 7 days, the same AII and AIII infusions were repeated. All patients showed blunted pressor responses to both AII and AIII before indomethacin and the responses were improved after indomethacin. Moreover, increment curves of blood pressure for AII were almost identical with those for AIII in individual patients both before and after indomethacin. ANG-(3-8) was infused iv in 3 normal men and 3 of the 4 patients with Bartter's syndrome at a rate of 3.500 pmol (2.838 ng)/kg X min for 15 min. Blood pressure rose in the normal men (12/12 mmHg on the average) but did not rise in the patients. These results suggest that AII, AIII and ANG-(3-8) have the same arteriolar binding sites in man. PMID- 2990135 TI - Opiomelanocorticotropic-like hormones in Vipera aspis pituitary. AB - By using immunohistochemical techniques (indirect immunofluorescence and PAP) adrenocorticotropic-like hormone (ACTH), alpha-melanocyte stimulating-like hormone (alpha-MSH), beta-lipotropic-like hormone (beta-LPH), and beta-endorphin like (beta-EP) have been localized in the pituitary gland of newborn and adult Vipera aspis. The immunolocalization have been compared with the classical histochemical staining and the data show that the opiomelanocorticotropic-like cells resembles the B3 cells of the current literature. In the intermediate lobe both in adults and in newborns the same cells exhibit immunoreactivity for all the peptides considered, while in the rostral region of the pars distalis a difference has been observed between adults and newborns. In the adults, the cells immunostain for ACTH, beta-LPH, and beta-EP but not for alpha-MSH while in the newborns, the same cells show immunoreactivity for ACTH, beta-LPH, alpha-MSH but not for beta-EP. It has been hypothesized that in Vipera during maturation the pars distalis and the intermediate lobe can differently process the common precursor molecule. PMID- 2990136 TI - Steroid response to ACTH and to ascorbinic acid during infusion of etomidate for general surgery. AB - The characteristics of the steroid response during an etomidate-fentanyl combination was explored in two groups of five patients undergoing major abdominal vascular surgery. Cortisol, androstene-dion, compound-S and ACTH were measured during surgery for three hours. In the fourth hour, an ACTH1-24 stimulation test was performed in one group and the steroid response was assessed. The other group received ascorbinic acid intravenously and the reaction of the adrenals was measured in the same way. The result indicates that etomidate causes a blockade of the adrenal corticosteroid synthesis at the site of the hydroxylation of cholesterol. This blockade is due to the interaction of the imidazole structure with the cytochrome p-450. Ascorbinic acid is able to overcome this blockade, indicating that cytochrome p-450 is involved in the ascorbinic acid metabolism which is obligatory for the hydroxylation of cholesterol. PMID- 2990137 TI - Ultrastructural and cytochemical studies of the fate of unsecreted collagen precursors after administration of colchicine to mice. AB - The administration of colchicine disrupts the normal organization of the Golgi complex and blocks the secretion of collagen precursors in periodontal ligament fibroblasts of the mouse. The fate of the unsecreted collagen precursors contained in Golgi-derived saccules and newly formed dense bodies was followed by electron microscopy. A progressive condensation of saccule content along with phase separation of electron-dense and electron-lucent material was observed. Fusion of saccules with dense secretory bodies gave rise to larger inclusions (zebra bodies; ZB) filled with a combination of electron-dense and electron lucent material. In some ZB, these materials appeared to polymerize into fibrillar units. The fibrillar units stained with silver methenamine like normal collagenous fibrils. These results suggest that unsecreted collagen precursors accumulate in vesicular compartments within which partial polymerization can occur. This finding may explain some reports of intracellular collagenous fibrils in fibroblasts of pathologically altered connective tissues. PMID- 2990138 TI - [Post-traumatic subarachnoid hemorrhage of the cauda equina]. AB - A case of subarachnoid hematoma of the cauda equina is described. The lesion developed in a patient receiving anti-coagulant treatment after a seemingly ordinary fall from a ladder. Neurosurgery achieved complete regression of the neurological defects in two weeks. The mechanism of development of the lesions is discussed. This form of complication of spinal trauma is fortunately exceptional. PMID- 2990139 TI - Different metabolism of vitamin D2 and vitamin D3 in epileptic patients on carbamazepine. AB - Serum concentrations of vitamin D metabolites were measured in 30 epileptic outpatients on monotherapy with carbamazepine before and during treatment with either vitamin D2 or vitamin D3, 4000 IU per day for 24 weeks. Vitamin D2 treatment increased the serum concentration of 25OHD2, but a corresponding decrease in 25OHD3 resulted in an unchanged serum value of total 25OHD. Vitamin D3 treatment increased the serum concentration of 25OHD3. The resulting serum level of 25OHD was consequently twice the level of that in the D2-treated group. The serum concentrations of the dihydroxy metabolites showed a similar difference between the 2 treatment groups. We conclude that treatment with vitamins D2 and D3 in the same doses produces considerably different serum concentrations of vitamin D metabolites. If the present findings can be extrapolated to normal subjects, it is important to consider more carefully which D-vitamin should be used, both with regard to therapy and supplementation. PMID- 2990140 TI - Ultrastructure of cerebellar capillary hemangioblastoma. III. Crystalloid bodies in endothelial cells. AB - The endothelial cells of three cases of cerebellar capillary hemangioblastoma were studied by means of electron microscopy. Crystalloid bodies, not previously described in the vessels of the central nervous system (CNS), were found in 5% 10% of the endothelial cells, more often in the capillaries with small irregular lumens. They were not observed in the pericytes or stromal cells. They were round to polygonal, 0.5-1.0 micron in size and composed of substructural units of parallel thick and thin electron-dense lines with a periodicity of 180-220 A. They coexisted occasionally with Weibel-Palade bodies but shared no structural correlation with them. The nature and significance of these crystalloid bodies are as yet unknown. Their close association with pinocytic vesicles suggests that the substructural crystalline might be the result of reconstruction of absorbed material from either the vascular lumen or the interstitium. Their subsequent development, associated with lysosomal activity, into intracytoplasmic dense bodies is postulated. Since crystalloid bodies were found only in endothelial cells, particularly those active in proliferation, they may play a pathophysiologic role in relation to angiogenesis. PMID- 2990141 TI - Neocortical cytopathology in focal aminopyridine seizures as related to the intracortical diffusion of [3H] 4-aminopyridine. Electrophysiologic and light microscopic studies. AB - Focal seizure was induced in rat and cat neocortex by the topical application of aminopyridines. The epileptic character of the focal events was followed by surface and intracellular recordings of seizure activity. The pathologic alterations in the neurons, the glial cells, and the protein permeability of the neocortical blood vessels were investigated by means of light microscopy, using standard histological stainings and immunohistochemical detection of endogenous serum albumin. Diffusion of [3H] 4-aminopyridine in the neocortex was studied by light-microscopic autoradiography. The spreading of the neuropathologic changes strictly followed the diffusion of the tritiated compound, suggesting the gradual involvement of neocortical layers in the seizure process. PMID- 2990142 TI - Immunohistochemical studies on cellular character of microtumors induced by ethylnitrosourea in the rat brain utilizing anti-Leu 7 and anti-glial fibrillary acidic protein antibodies. AB - To clarify the chronologic changes in the cellular morphology of ENU-induced rat brain tumors, microtumors in the early stage were examined immunohistochemically in comparison with macrotumors in the advanced stage. The tumor cells composing microtumors were negative for glial fibrillary acidic protein (GFAP), a specific marker of astrocytic cells, and Leu 7, a marker of oligodendrocytes, while cells of macrotumors were positive for either GFAP or Leu 7, showing characteristics of mature glial cells. The results suggested that the small round cells in the early developmental stage, generally thought to resemble mature oligodendrocytes, are not differentiated oligodendrocytes or astrocytes. PMID- 2990143 TI - Cortical cerebellar degeneration with testicular neoplasm. AB - Cortical cerebellar degeneration was found in a 28-year-old man with testicular neoplasm. The patient, who had undergone a left orchidectomy for the testicular tumor, developed progressive cerebellar symptoms with mental changes 7 months later. The autopsy revealed the spread of a malignant germ cell tumor of the testis, and cortical cerebellar degeneration in the central nervous system (CNS) which was characterized by almost complete loss of Purkinje cells and degeneration of the both dentate nuclei and superior cerebellar peduncles. The present case is the first of cortical cerebellar degeneration combined with testicular neoplasms. PMID- 2990144 TI - Difference of lectin binding sites of secretory granules between normal pituitary and adenoma cells. AB - Electron-immunocytochemical staining with lectin (concanavalin A: Con A) binding sites analysis was applied to study secretory granules of human pituitary adenomas and surrounding normal pituitary tissue using post-embedded serial ultrathin sections. Twelve cases of human pituitary adenoma and three specimens of normal pituitary tissue surrounding adenomas were studied: the cases were operated on between 1982 and 1984. The tumors consisted of four prolactin (PRL)-, six growth hormone (GH)-, and two adrenocorticotropic hormone (ACTH)-producing adenomas. In parallel with the detection of Con A binding sites of secretory granules, their secreting hormones were characterized electron-microscopically with the immunocytochemical horseradish peroxidase (HRP) labeling using the avidin-biotin technique. The two cases of ACTH-producing adenomas showed either weak or negative reactions with Con A on secretory granules, while normal ACTH producing pituitary cells showed strong reactions with Con A on every secretory granule observed. Large secretory granules of PRL- or GH-producing cells showed negative reactions with Con A both in the pituitary adenoma and normal pituitary, while some small granulated or sparsely granulated adenoma cells also showed strong reactions with Con A. The complexity of human pituitary adenomas is illustrated as well as the difference in biochemical structure of normal pituitary cells and pituitary adenoma cells secreting the same specific hormone. PMID- 2990145 TI - Neuropathologic observations in acquired immunodeficiency syndrome (AIDS). AB - Neuropathologic changes are described in eight fatal cases of the acquired immunodeficiency syndrome (AIDS), including four subjects who were i.v. drug abusers (two women, two men), two Haitian men, one bisexual man, and one women who had no known risk factors for AIDS. All cases had significant infections of the brain, with examples of bacterial, fungal, protozoal, and viral disease in the group. Combined infections were observed in three cases, including one case of previously unreported central nervous system (CNS) abscess due to Nocardia (combined with Salmonella). The most frequent CNS pathogen was Toxoplasma gondii, which in both Haitian subjects co-existed with systemic tuberculosis, leading to diagnostic difficulty. Only one case of "subacute encephalitis" was represented, although in other cases microglial nodules were associated with Toxoplasma and Cryptococcus organisms, as well as probable cytomegalovirus. Non-infectious complications included a cerebral infarct (one case), central pontine myelinolysis (one case), and diffuse microcystic change of the white matter (one case). PMID- 2990146 TI - Effect of chlorpromazine on the localization of cAMP phosphodiesterase. AB - Chlorpromazine (CPZ) at dosages of 10 mg/kg body weight (b.wt.) affected the cytochemical localization of cAMP-dependent phosphodiesterase (cAMP PDE) activity in the synapses of the rat frontal cortex. Postsynaptic cAMP PDE activity was inhibited, and presynaptic activity increased. CPZ also inhibited membrane-bound ATPase activity in the frontal cortex. The activity of Na+-K+-ATPase was significantly (P less than 0.005) inhibited in isolated plasma membranes from the rat frontal cortex. CPZ exposure also affected the cytochemical localization of cations with potassium pyroantimonate. Precipitate, which could be removed with 5 mm EGTA, was decreased in the mitochondria and synaptic vesicles in presynaptic areas after CPZ treatment. The incorporation of 45Ca2+ into slices of the rat frontal cortex was also significantly (P less than 0.001) inhibited by CPZ. This ultrastructural study shows that CPZ may affect biochemical events in an opposite manner in the pre- and post-synaptic areas of some neurons of the frontal cortex. PMID- 2990148 TI - Astrocyte differentiation induced by Junin virus in rat brain cell cultures. AB - Morphological and immunocytochemical differentiation was observed in astroglial cell cultures of the rat infected with Junin virus. From days 3 to 6 postinoculation (p.i.), GFAP immunostaining was observed in both the perikaryon and processes of maturated astrocytes, whereas it was limited to the perikaryon in less differentiated cells. The rather slow spontaneous differentiation usually occurring in astroglial cell cultures was seen to be accelerated by viral infection, mimicking the astrocytic reaction formerly described in Junin virus inoculated mice. Infected cell monolayers showed orderly development, maintenance of contact inhibition, and exhaustion of cell cultures beyond the 6th-7th passages. The morphological and immunocytochemical maturation effects of Junin virus on astroglial cells were evident, but to a significantly lesser degree than those caused by rat brain extract. The glial cell cultures proved a valuable tool for the study of virus-cell interaction, since the immune response and the structural complexity of the whole animal can thus be avoided. PMID- 2990147 TI - Characterization of four human malignant glioma cell lines. AB - In this paper, the characterization of four human malignant glioma cell lines is described. The four lines are positive for glial fibrillary acidic protein (GFAP) in variable amounts. One of them, LN 992, is positive for S-100 protein. Myelin basic protein could not be detected in any of the four lines. The four lines had high levels of CNPase activity. The karyotype shows polyploidy for all lines, with modal numbers ranging from 80 to 120 and various numbers of marker chromosomes. Particular attention has been paid to the surface phenotype and a panel of three antiglioma monoclonal antibodies (Mabs), five antimelanoma Mabs, one anti-CALLA Mab, and two anti-HLA-DR Mabs has been used in an antibody-binding radioimmunoassay for the four cell lines. Lines LN 215 and LN 235 are positive with two antiglioma Mabs, LN 992 is negative. The four lines are positive with all five antimelanoma Mabs, except for LN 992 which ist negative with Mab D5. LN 992 and LN 215 are positive with the anti-CALLA Mab N2A12. LN 308 and LN 992 are positive with anti-HLA-DR Mab D4-22. There was no correlation between the in vitro morphology of the lines and the expression of the various biochemical or surface markers. These results stress the heterogeneity of the phenotype of human malignant glioma lines. These lines will be useful tools for further immunologic studies. PMID- 2990149 TI - Spinal ganglia and peripheral nerves from a patent with Tay-Sachs disease. Morphological and ganglioside studies. AB - We examined the spinal ganglia and peripheral nerves from a patient with Tay Sachs disease, comparing the other nervous tissues morphologically and lipid biochemically. The spinal ganglia and peripheral nerves showed numerous membranous cytoplasmic inclusion bodies (MCBs), which are characteristic of GM2 ganglioside storage in the neuronal cell bodies of the patient brains. In spinal ganglia, all neurons and satellite cells around the neurons contained membrane bound lipid materials. In peripheral nerves, Schwann cells and myelinated axons except for enlarged axons were devoid of MCBs. Major ganglioside stored in both spinal ganglia and peripheral nerve was also GM2-ganglioside. The contents of ganglioside in the spinal ganglia and peripheral nerves were 50 and 10 times more than those from normal tissues, respectively. The spinal cord contained a slightly higher amount of gangliosides than the normal control. The cerebral white matter totally demyelinated in this patient accumulated a much higher amount of gangliosides than the cerebral gray matter. The retinal tissue showed GM2-ganglioside as the major one also. PMID- 2990151 TI - A retromedullary arteriovenous fistula associated with the Klippel-Trenaunay Weber syndrome. A clinicopathologic study. AB - An intraspinal vascular malformation associated with the Klippel-Trenaunay-Weber (KTW) syndrome initially was thought to be intramedullary on angiographic findings. Postmortem examination revealed an entirely posterior extramedullary arteriovenous fistula (AVF) fed by the anterior spinal artery. The association of the KTW syndrome with a so-called intramedullary AVF has been described in the literature without any pathologic confirmation (11 cases). Our case emphasizes the difficulty of determining the exact morphology and location of spinal AVF on arteriography. The association of the KTW syndrome with a retromedullary AVF can be explained on a developmental basis. PMID- 2990150 TI - Peripheral nervous tissue injury induced by galactocerebroside and galactocerebroside immune complexes. AB - It was demonstrated that New Zealand Albino rabbits sensitized to galactocerebroside had high levels of anti-galactocerebroside antibody and of immune complexes. The rabbits was high titers of immune complexes developed demyelination in the peripheral nerves. Lesion were produced in the peripheral nerves of mice by the i.m. injection of galactocerebroside immune complexes. The lesions were characterized by axonal degeneration, infiltrating macrophages containing myelin debris, and an inflammatory infiltrate of polymorphonuclear and mononuclear cells. Rabbit immunoglobulin and mouse C3 were observed around the endoneural blood vessels. These results suggest that galactocerebroside immune complexes may play a role in the pathogenesis of mouse peripheral nerve lesions, due to the production of vasculomyelinopathy. PMID- 2990152 TI - Cochlear changes after herpes simplex virus infection. AB - After direct inoculation of herpes simplex virus (HSV) into the scala tympani of the guinea pig, the tectorial membrane showed various morphological changes: atrophy, roll-up and dot formation. Immunofluorescent and electronmicroscopic studies revealed that the changes were due to HSV infection. The findings were compared with those observed in the temporal bones of a 77-year-old patient who suffered from sudden deafness. The tectorial membranes of both temporal bones showed various changes identical with those observed in experimental viral labyrinthitis. This supports the view that sudden deafness in this particular patient was of viral origin. In the animal experiment, HSV antigen could be detected from the cochlea of the non-inoculated side, which was morphologically normal. Further study is required to reactivate HSV in the cochlea with latent infection. This animal can probably be used as an animal model for sudden deafness. PMID- 2990153 TI - Analysis of trace elements in the human labyrinth by using proton microprobe. AB - The concentration of zinc in the spiral tract is age-dependent. The accumulation of zinc starts at the bone surface towards the fundus of the internal auditory meatus and continues throughout the whole thickness of the separating wall to reach in advanced age the bone surface towards the basal coil of the cochlea. The concentration of calcium decreases with age. These findings prove our previous histological and densitometric as well as macroscopical investigations of this region. PMID- 2990154 TI - Establishment and characterization of cell lines from human small cell and large cell carcinomas of the lung. AB - Five new small cell carcinomas (SCC) cell lines and a large cell carcinoma (LCC) cell line were established from human lung cancers. The SCC cell lines had, as a group, common phenotypic properties which distinguished them from non-SCC cell lines. However, the studies also revealed a considerable biological heterogeneity among the individual SCC cell lines. Thus, the SCC cell lines had a typical growth pattern with cell clusters in suspension or partly adherent to the bottom. All the lines examined grew in agarose with variable cloning efficiencies, and all but one line formed tumors subcutaneously in nude mice. The ultrastructure of the SCC cell lines was characteristic with dense core granules at a variable frequency. Neuron-specific enolase was detectable in all SCC cell lines, usually in large amounts, and an inconstant production of a spectrum of polypeptide hormones was found, typical of SCC. The LCC cells proliferated in monolayers, formed colonies in agarose and grew in nude mice. Ultrastructurally, the LCC cell line differed from the SCC cell lines in having intra- and intercellular lumina and tonofilaments. The capacity of the LCC and a previously established squamous cell carcinoma cell line (U-1752) to produce neuron-specific enolase and polypeptide hormones was characteristically much lower than that of the SCC cell lines. We conclude from this study that SCC cell lines, although individually distinct from one another, are quite homogeneous as a group in expressing a set of basic common neuro-endocrine markers. However, these studies also suggest some biological relationship between SCC, LCC and SQC by virtue of their expression of some common neuro-endocrine markers, in support of the concept of a common histogenetic origin of human lung cancers. PMID- 2990155 TI - Evaluation of two commercial kits for rapid detection of human rotavirus in feces: Rotalex, a latex agglutination test and Rotavirus ELISA Kit. AB - 96 fecal specimens from children with symptoms of gastroenteritis were tested with a latex agglutination test (Rotalex, Orion Diagnostica, Finland) and an enzyme-linked immunosorbent test (Rotavirus ELISA Kit, Dakopatts, Copenhagen, Denmark). Using the ELISA method as reference, the latex agglutination (LX) came out with three non-specific and two false negative results. The LX test was found to be a rapid, practical and non-expensive choice for a laboratory examinating few samples daily. PMID- 2990156 TI - Variations in binding of mammalian fibrinogens to streptococci groups A, B, C, E, G and to Staphylococcus aureus. AB - Twenty-eight beta-hemolytic streptococci of groups A, B, E, G and Streptococcus equisimilis as well as four Staphylococcus aureus strains were tested for their ability to bind fibrinogen preparations from different animal species: homo, baboon, rabbit, rat, guinea-pig, dog, horse, pig, cow and sheep. The patterns of binding indicated differences in the structures of the bacterial fibrinogen receptors. There were higher binding levels in streptococci groups A, G, and S. equisimilis than in representative group B and E strains. Considerable differences in the binding capacity were found within streptococci groups A and E. Group C and group G strains showed rather similar patterns and could be further divided into high-level and low-level binding strains. There is no correlation between binding levels of different animal fibrinogen preparations and the strains isolated from corresponding animals. Recent studies by others have shown that resistance to phagocytosis is mediated by fibrinogen-binding in streptococci group A. The existence of similar fibrinogen-binding structures in several streptococcal species indicates an important role with a definite survival value. It also suggests that M or T protein analogues are present in streptococci groups C, G and E. PMID- 2990157 TI - Phenotypic and genotypic characterization of penicillinase-producing strains of Neisseria gonorrhoeae. AB - Thirty penicillinase-producing Neisseria gonorrhoeae (PPNG) strains, 26 representing 9 serovars of the protein IB and 4 representing 2 serovars of the protein IA serogroup, had 10 different patterns when investigated by the restriction enzyme (RE) technique with Hind III enzyme. One RE pattern dominated, representing 11 strains with 6 different serovars of both the protein IA and IB serogroup. Identical serovars and RE patterns were found among 7 patients (3, 2 and 2, respectively) with positive epidemiology. The majority of the strains (24/30) were resistant to, or showed decreased sensitivity to, doxycycline with minimum inhibitory concentrations between 1 and 8 mg/l, consistent with the Far East origin. All the 30 PPNG strains showed by agarose gel electrophoresis to harbour a 2.8 Mdalton, a 4.4 Mdalton, as well as a 24.5 Mdalton plasmid, whereas the two non-PPNG strains had only the 2.8 Mdalton plasmid. PMID- 2990158 TI - Steric aspects of agonism and antagonism at beta-adrenoceptors: experiments with the enantiomers of clenbuterol. AB - The enantiomers of clenbuterol, a beta 2-selective adrenoceptor agonist with partial agonistic activity, were examined with respect to their ability to react in vitro on adrenoceptors in the trachea (mostly beta 2), the soleus muscle (beta 2) and in the papillary muscle of the left ventricle (beta 1) from the guinea pig. (-)-Clenbuterol relaxed the carbachol contracted trachea and depressed the subtetanic contractions of the soleus muscle in a concentration-dependent manner. (+)-Clenbuterol was at least 1,000 times less potent in this respect. Both isomers inhibited competitively the effect of isoprenaline on the trachea, the ( )-isomer being about 100 times more active than the (+)-isomer. None of the isomers showed any detectable positive inotropic effect on the papillary muscle but both inhibited competitively the response to isoprenaline. Also in this respect (-)-clenbuterol was more potent than (+)-clenbuterol. It is concluded that the beta 2-agonistic as well as the beta 1-antagonistic effect of clenbuterol resides in the (-)-isomer and that the (+)-isomer does not seem to contribute to the pharmacological effects displayed by racemic clenbuterol. PMID- 2990159 TI - Effect of thyroxine on hepatic glucose-6-phosphatase activity and glycogen content of toad (Bufo melanostictus) and Lata fish (Ophicephalus punctatus) at different stages of life. AB - The effects of thyroxine (T4) on hepatic glucose-6-phosphatase activity and glycogen content in toad (Bufo melanostictus) and Lata fish (Ophicephalus punctatus) were studied in order to show the difference, if any in the enzyme activity and glycogen metabolism in their liver. Thyroxine injections (1 microgram/g) for five consecutive days caused a reduction in hepatic glucose-6 phosphatase activity and glycogen content in toads of immature, juvenile and adult stages. In contrast, Lata fish of different stages showed an enhancement of hepatic glucose-6-phosphatase activity after T4 treatment (1 microgram/g, 5 injections). The liver glycogen content in Lata fish of different age groups was found to be reduced after T4 injections, but not so much as in the toad. PMID- 2990160 TI - Effect of dehydroepiandrosterone sulfate on uterine cervical ripening in late pregnancy. AB - In order to elucidate the effects of dehydroepiandrosterone sulfate (DHAS) on softening and dilatation of the uterine cervix, changes of oestriol, 17 beta oestradiol and progesterone levels in serum and cervix, Bishop score and collagenase activity in the cervical tissue were assessed in pregnant women before and after treatment with DHAS. 17 beta-oestradiol level in the serum and cervical tissue markedly increased after the administration of DHAS, while oestriol level remained unchanged. Serum progesterone level did not change in the majority of cases, while it decreased within several hours in patients in whom delivery was accomplished within 24 hours after the administration of DHAS. Among the factors connected with the Bishop score, effacement and consistency of the cervix were remarkably improved by DHAS administration. Total collagenase activity in the cervical tissue of patients treated with DHAS was elevated by an average of 152%. These results suggest that DHAS is potent in ripening the uterine cervix through an activation of collagenase activity induced by the enhanced conversion to 17 beta-oestradiol. Thus, DHAS administration in the late stage of pregnancy is valuable in prepartal treatment for induction of labour. PMID- 2990162 TI - Betamimetic effects on the electro-mechanical characteristics of the pregnant and postpartum myometrium in vitro. AB - Myometrial strips were studied in the two extremes of regulatory conditions by measuring isometric tension and resting potentials in vitro. The effects of isoproterenol (IP) and PGF2 alpha were studied on pregnant and postpartum rabbit uteri. Lower IP concentrations inhibited the activity of the pregnant uterus as compared to the postpartum myometrium. Both pregnant and postpartum myometrium could be hyperpolarized by IP, however, lower concentrations were needed during pregnancy. The inhibiting effect on mechanical activity of IP developed without any hyperpolarization at its lower concentration. PMID- 2990161 TI - Regulation of alpha- and beta-adrenergic receptor subclasses by gonadal steroids in human myometrium. AB - Both alpha- and beta-adrenergic receptors have been identified in the human myometrium by radioligand binding. Both adrenergic receptor subclasses have been shown to mediate the contractile response of the uterus upon catecholamine stimulation: alpha-adrenergic receptors cause uterine contraction while beta adrenergic receptors induce relaxation. We have identified alpha 1- and alpha 2 adrenergic receptors in myometrial membranes using the newly developed radiolabelled specific antagonists [3H]-prazosin and [3H]-rauwolscine. This enabled us to characterize both receptor subclasses individually. Beta adrenergic receptors were identified using the radiolabelled antagonist (-)-[3H] dihydroalprenolol. Binding of radioligands to the myometrial membrane receptors was rapid, readily reversible, of high affinity and stereoselective. The total number of alpha 1-, alpha 2- and beta-receptors was determined by Scatchard analysis of radioligand saturation binding and the beta/beta 2-receptor ratio was determined by computer analysis of the beta 2-selective antagonist ICI 118 551) ( )-[3H]-dihydroalprenolol competition binding curves. This enabled us to study the regulation of both alpha- and beta-receptor subclasses under various physiological and pharmacological conditions in the human, i.e., during different phases of the menstrual cycle, in postmenopausal women and during depo-progestin (Medroxyprogesterone acetate) therapy. Only the alpha 2- and beta 1-adrenergic receptor concentrations were found to be subjected to gonadal steroid regulation. The number of alpha 2- and beta 1-adrenergic receptors increased concomitantly with circulating plasma oestradiol levels. This effect was counteracted by progesterone. The number of alpha 1- and beta 2-adrenergic receptors was unaffected by the gonadal steroid environment. These results are an example of the heteroregulation of membrane receptors by oestrogens and progesterone and cast new light on the regulatory mechanisms involved in uterine contractility in the human. PMID- 2990163 TI - Characterization of beta-adrenergic receptors in human myometrium and placenta. AB - The binding characteristics of the beta-adrenergic ligand [3H]-dihydroalprenolol (DHA) were determined in particulate membranes of early and term human placenta and myometrium. Human placenta is rich in beta adrenergic receptors and the binding capacity increases with increasing gestation. There was no difference in receptor density in human myometrium comparing the first trimester and term. Binding of DHA to human myometrial beta-adrenergic receptors provides a tool for the analysis of beta-adrenergic agents as potentially useful tocolytic agents. PMID- 2990164 TI - Further evidence for the existence of interactions between receptors for dopamine and neurotensin. Dopamine reduces the affinity and increases the number of [3H] neurotensin binding sites in the subcortical limbic forebrain of the rat. PMID- 2990165 TI - Capsaicin treatment decreases tissue levels of neurokinin A-like immunoreactivity in the guinea pig. PMID- 2990166 TI - Changes in growth hormone binding and metabolic effects of growth hormone in rat adipocytes following hypophysectomy. AB - Growth hormone (GH) binding and the effect of GH and insulin on glucose metabolism in rat adipocytes were studied at various time periods following hypophysectomy. Male rats were hypophysectomized at 33-34 days of age. After 6 h, 20 h or 3, 7 and 14 days adipocytes were prepared from epididymal fat pads by mild collagenase digestion (0.5 mg X ml-1, 60 min, 37 degrees C). Glucose metabolism was studied by determining the production of CO2 from [14C]glucose and the incorporation of [14C]glucose into lipids. GH binding was measured in cell aliquots using [125I]hGH. No difference in GH binding to adipocytes was observed between control rats and rats hypophysectomized or sham-operated 6 h earlier. GH binding was significantly decreased 20 h after hypophysectomy and declined further with time after hypophysectomy. Adipose tissue from normal rats is usually refractory to the insulin-like effect of GH. Adipocytes isolated from normal rats were, however, usually responsive to GH immediately after cell isolation, suggesting that refractoriness to the insulin-like effect of GH was lost during the time required for the preparation of adipocytes. The magnitude of the response to GH in adipocytes progressively declined with time after hypophysectomy. The decreased responsiveness to GH with time after hypophysectomy parallelled the decrease in GH binding. The results suggest that the pituitary, directly or indirectly, is necessary for the maintenance of GH binding sites in adipose tissue and that these binding sites are related to the insulin-like effect of GH. PMID- 2990167 TI - Adenosine analogues stimulate cyclic AMP formation in rabbit cerebral microvessels via adenosine A2-receptors. AB - This study has examined the accumulation of cyclic AMP in microvessels from rabbit and feline cerebral cortex induced by a series of adenosine analogues to determine the type of receptor involved. The conversion of tritium labelled adenine nucleotides to [3H]cyclic AMP was determined in [3H]adenine labelled microvessels in the presence of an inhibitor of cyclic AMP hydrolysis, rolipram (30 microM). In microvessels from both cats and rabbits two adenosine analogues, N-5'-ethylcarboxamido adenosine (NECA) and L (S)-phenylisopropyladenosine (PIA), stimulated cyclic AMP accumulation. The response was larger and more reproducible in rabbits than in cats. In rabbit cerebral microvessels the order of potency as stimulator of cyclic AMP accumulation was NECA greater than 2-chloroadenosine greater than L-PIA greater than cyclohexyladenosine (CHA) greater than D-PIA. This order of potency defines the receptor involved as being of the A2 subtype. Although the maximal response to CHA appeared to be lower than that to NECA, CHA did not inhibit the response to NECA, suggesting that it is not a classical partial agonist. In the presence of the adenylate cyclase stimulating compound forskolin (I microM) NECA was more active than in its absence (close to 30-fold increase in EC50) and also produced a maximal effect six times higher. The maximal responses to PIA and CHA increased proportionally in the presence of forskolin. These results show that rabbit cerebral microvessels possess adenosine receptors of the A2 subtype capable of stimulating the formation of cyclic AMP. The functional significance of such receptors is not known, but may be related to regulation of vascular permeability. PMID- 2990168 TI - Comparison between the steroidogenic responses after pulsatile and continuous administration of gonadotrophin releasing hormone to superfused rat follicles. AB - Gonadotrophin releasing hormone (GnRH) exhibits both stimulatory and inhibitory effects on the ovarian tissue in the rat. For the action of GnRH on the pituitary, the mode of administration is of utmost importance. The binding characteristics and regulation of the GnRH receptors found in the ovary are very similar to those of the pituitary. The aim of the present study was to compare the steroidogenic response of superfused, preovulatory rat follicles to continuous and pulsatile administration of a GnRH analogue (GnRHa). The results obtained with a single pulse of luteinizing hormone (LH) served as a control. Repeated pulses (1 pulse X h-1) or continuous administration of GnRHa was found to be more effective than a single pulse of GnRHa. The effect of GnRHA, irrespective of the mode of administration, was most pronounced on the secretion of 20 alpha-OH-progesterone (20 alpha-OHP). Also the release of testosterone (T) and oestradiol (E2) was stimulated but to a lesser extent. In comparison to LH, the GnRHa response was delayed and without detectable release of cyclic AMP (cAMP). These results illustrate that pulses of GnRH stimulate the steroidogenesis in preovulatory follicles. In contrast to the effects of GnRHa on the pituitary, the modes of administration seem to be of less importance for the stimulation of the rat ovary. PMID- 2990169 TI - Prefrontal cortex dysfunction as a common factor in psychosis. AB - It has recently become possible to arrive at a testable biopsychological model, according to which a dysfunction of the prefrontal cortex is the common factor in psychotic disorders; if persistent, this leads to a dissolution of the highest level of brain function, in the sense of Hughlings Jackson. The model is in agreement with newer findings in normal and pathological physiology and can serve as basis for specific experimental questions. It is compatible with a unitary epistemological viewpoint. PMID- 2990170 TI - Repeated computed tomography during treatment of small cell carcinoma of the lung. AB - Twenty-three patients with small cell carcinoma of the lung were examined with CT and conventional chest radiography before treatment, 19 after 3 courses of chemotherapy, before radiation therapy, and 8 after cessation of chemotherapy and radiation therapy. In most patients high abdominal CT was also employed. CT provided more information about the thoracic involvement than did conventional radiography in more than half of the cases. In 3 cases CT before irradiation demonstrated progression in the thorax, not visible at conventional radiography, and in 4 cases progression of extrathoracic metastases in spite of intrathoracic regression. After therapy, one case of progression in the thorax not seen at conventional radiography was demonstrated. With repeated CT examinations of the thorax and abdomen, progression/regression both intra- and extrathoracically is well demonstrated. PMID- 2990171 TI - Significance of collateral arterial supply to Wilms' tumours. AB - The presence of collateral arterial supply was examined by angiography in 19 children with Wilms' tumour. Collateral arterial supply was related to tumour size. Ten of 14 tumours displaying collateral circulation were entirely intrarenal at operation, confirmed by histopathology. Angiography in Wilms' tumour is indicated when the results of urography, ultrasonography or computed tomography are equivocal or extrarenal tumour growth is suggested. PMID- 2990172 TI - Hypertension in pregnancy. Long-term effects on blood pressure in mothers and children. PMID- 2990174 TI - [Intraparenchymal abscess in a renal graft]. PMID- 2990173 TI - [Dopaminergic disequilibria in hemicrania]. PMID- 2990175 TI - When should we request radionuclide brain scans in elderly patients? AB - The radionuclide brain scans performed on a gamma camera (GC scans) in 146 patients over 65 years of age during a 6-month period are reviewed. Forty-four scans (30.1%) were reported as showing an abnormality. Three patients subsequently underwent surgery and drug therapy was commenced in 11 cases. However, in most cases the GC scan proved unhelpful in the patients' management. The usage and role of GC scanning in elderly patients admitted with a neurological disorder is examined. PMID- 2990176 TI - H+ production by isolated cells from human gastric mucosa. AB - Gastric mucosal cells were isolated from human mucosa obtained at surgery. H+ production was indirectly estimated by 14C aminopyrine (AP) uptake. The maximal response to histamine occurred after 30 min of incubation whereas intrinsic factor (IF) secretion was maximal after only 7.5 to 15 min. According to the concentration response curve 10(-4) mol/1 histamine proved to be the most effective concentration, the response to which was completely inhibited by ranitidine. Carbachol, dibutyryl cAMP and IMX also enhanced AP uptake, IMX being even more powerful than histamine. Carbachol and IMX failed to potentiate the response to histamine. Parietal cell fractions enriched by a Percoll density gradient revealed a pronounced background stimulation so that additional stimulation by test agents was less effective than in non-fractionized cells. PMID- 2990177 TI - Histamine may act through cyclic AMP on hippocampal neurones. AB - Bath applied histamine and 8-bromo-cyclic AMP and intracellularly injected cyclic AMP block-long-lasting after-hyperpolarizations and the accommodation of firing in CA1 pyramidal cells recorded in rat hippocampal slices. This action is due to reduction of a calcium-activated potassium conductance (gK(Ca] and leads to potentiation of excitatory signals including epileptiform discharges. The effects are further potentiated and prolonged by a phosphodiesterase inhibitor (Ro 20 1724). PMID- 2990178 TI - Evidence for the role of cAMP-dependent protein kinase in the down-regulation of hypothalamic HD: reversal of cAMP-(ATP) induced inhibition of HD activity by the 'Walsh' inhibitor of cAMP-dependent protein kinase and by cyclic GMP. AB - Under the total blockade of PDE1 and the presence of endogeneous ATP and MgCl2, the inhibitory effect of cAMP on HD activity could be demonstrated as low as 8.7 X 10(-8) M concentration in a 20,000 g supernatant of a sustained homogenate of rat hypothalamus. A total reverse of this action and also a partial release of the cAMP-induced inhibition of HD, occurred at higher concentrations of cAMP, and ATP could be achieved by an endogeneous inhibitor of cAMP-dependent protein kinase or by cyclic GMP. The reversal of cAMP action by PKI seems to serve a strong evidence for the role of cAMP-dependent protein kinase (EC 2.7.37: ATP protein phosphotransferase) in this action and emphasized the involvement of a direct or an indirect phosphorylation in the regulation of HD activity. The stimulatory effect of cyclic GMP on cAMP-induced inhibition of HD or its 'direct' effect on histamine formation is asserted, probably through the activation of PDE, or through independent stimulatory machinery, coupled to the cyclic GMP system. PMID- 2990179 TI - Surveillance following orchiectomy for clinicaL stage I testicular germ-cell malignancy. PMID- 2990180 TI - [Radiotherapy of testicular tumors]. PMID- 2990181 TI - Management of germ cell tumors of the adult testis at the Memorial Sloan Kettering Cancer Center. PMID- 2990182 TI - The Danish Testicular Carcinoma Study. Results from recent clinical trials in non seminomatous tumours. PMID- 2990183 TI - Personal experience in the management of germinal testis tumors. PMID- 2990184 TI - Nonseminomatous germ celL testicular tumors. Review of the UCL experience. PMID- 2990185 TI - Treatment of residual intrathoracic metastases in metastatic tumors. PMID- 2990186 TI - Human T-cell leukemia/lymphoma syndrome. AB - This syndrome should be suspected in patients with clinical features of lymphadenopathy, hepatosplenomegaly, hypercalcemia, bone lesions and circulating lymphocytes with pleomorphic nuclei. Most biopsy material has morphologic characteristics of intermediate or high-grade non-Hodgkins lymphoma. Antibody titers to human T-lymphotropic virus type I confirm the diagnosis. Treatment with combination chemotherapy results in remission for most patients, but duration of response is usually short. PMID- 2990187 TI - Adrenal function following coronary bypass surgery. AB - Little is known about adrenocortical function after coronary bypass surgery in which moderate to deep hypothermia and cardiopulmonary bypass are used particularly with intraoperative steroid administration. Therefore, we performed a pilot study in which immediately preoperative and 18-hour postoperative serum cortisol levels were determined in eight patients who received 1.0 to 1.5 gm of methylprednisolone intravenously during surgery; postoperative serum cortisol (3 +/- 1 microgram%) levels were lower than preoperative levels (15 +/- 3 microgram%, p less than 0.05). To determine the possible cause of these striking findings, the effects of moderate to profound hypothermia and cardiopulmonary bypass upon adrenocortical functioning were investigated without the influence of intraoperative steroid administration. Serum cortisol and aldosterone levels and their response to adrenocorticotropic hormone (ACTH) (Cortrosyn) were determined before coronary bypass surgery and at various postoperative intervals in seven patients. Postoperative cortisol and aldosterone levels increased markedly over their preoperative values, reaching a maximum at 6 to 12 hours (cortisol 16 +/- 8 vs 63 +/- 23 micrograms%, p less than 0.05, aldosterone 15 +/- 5 vs 51 +/- 22 ng%, p less than 0.05). Adrenal response to ACTH was normal preoperatively, during rewarming from hypothermia, and 18 hours, and 7 days postoperatively. In summary, normal adrenal responsiveness occurs after coronary bypass surgery, in spite of hypothermic cardiopulmonary bypass and the effects of anesthesia, and a single dose of methylprednisolone during surgery is associated with markedly lower serum cortisol levels and prevents the usual adrenal stress response to bypass surgery for at least 18 hours postoperatively. PMID- 2990188 TI - A stable isotope study of copper absorption in young men: effect of phytate and alpha-cellulose. AB - A sixty-three day study was conducted with young men confined to a metabolic unit to study the effects of alpha-cellulose and phytate on copper absorption. Copper absorption was determined with 65Cu, a stable isotope of copper, during each of 3 dietary treatments (basal diet, basal diet + alpha-cellulose, or basal diet + phytate). The addition of alpha-cellulose or phytate to the basal diet did not affect copper absorption. Average copper absorption was 35.0% from the basal diet, 34.1% from the diet with 0.5 g alpha-cellulose per kg body weight added, and 31.4% from the diet with 2.34 g of phytate as sodium phytate added to the diet. Copper absorption was significantly different between subjects and averaged 44.1%, 26.8%, 33.4%, and 29.5% in individual subjects. The results suggest that high levels of either alpha-cellulose or phytate do not have marked effects on copper absorption, but copper absorption differs between individuals. PMID- 2990189 TI - Dietary fiber effects on passage rate and breath hydrogen. AB - Fermentation of fiber and passage to the large intestine were monitored by measuring hydrogen gas (H2) concentration in expired breath. Five subjects consumed meals containing no added fiber or 40 g of wheat bran (WB), corn bran (CB), oat bran (OB), or citrus flour (CF) replacing white flour. Breath samples were obtained at 30 min intervals for 9 h after the test meals. Mean H2 concentration (ppm), averaged over the 9 h test period, ranged from 7.5 (CB) to 12.0 (OB) and tended to increase with addition of OB, CF, or WB but decrease with addition of CB. Hours from the meal to the highest H2 peaks were 4.7 (WB), 5.6 (CB), 6.2 (OB), 6.4 (CF), and 8.2 (basal). The gel-like fibers (OB and CF) resulted in greater H2 concentration and less acceleration of passage rate than the particulate fibers (WB and CB). PMID- 2990190 TI - Microcystic adenoma of the pancreas with myoepithelial cells. A hitherto undescribed morphologic feature. AB - A case of microcystic adenoma (glycogen-rich cystadenoma) of the exocrine pancreas in a 62-year-old white female presenting with adult-onset diabetes mellitus is reported. Clinical, histopathologic, and ultrastructural findings are discussed with particular emphasis on the presence of myoepithelial cells, a morphologic feature not seen in this tumor previously. The controversy over diabetes mellitus as a significant related factor is also discussed. PMID- 2990191 TI - Identification of complex phenotype (OKT4+/OKT8+) on adult T-cell leukemia cells by sequential application of indirect rosette assay with protein A-coated ox red blood cells and immunoperoxidase technic. AB - The specific immunophenotypic characterization of the hematologic malignant diseases has been obtained most commonly by the immunofluorescent technic. It requires a microscope with illuminator or the expensive flow cytometer that usually precludes morphologic assessment. It was compared with the indirect rosette assay and the immunoperoxidase study, which allowed both immunologic and morphologic assessment. These three technics appeared to be comparable quantitatively, and the latter two technics utilizing cytospin preparations were applied sequentially to the leukemic cells of adult T-cell leukemia with a complex phenotype (OKT4+/OKT8+) and successfully displayed the double markers on each leukemic cell. PMID- 2990192 TI - Terminal duct carcinoma of minor salivary glands. A nonpapillary subtype of polymorphous low-grade adenocarcinoma. AB - The clinicopathologic features of five terminal duct carcinomas arising in minor salivary glands are presented. These nonpapillary, low-grade adenocarcinomas are part of the spectrum of polymorphous low-grade adenocarcinoma. The patients ranged in age from 47 to 77 years. Symptoms were nonspecific and related to a mass that usually was present for several years. Histologically, the tumors had a ductal pattern with areas of solid and cribriform architecture. Cytologically, they were composed of uniform, cuboid cells with round to oval nuclei and fine chromatin. One patient developed a recurrence after 13 years, but none of the tumors has metastasized. One lesion in this series was studied ultrastructurally and demonstrated both glandular and pseudoglandular structures analogous to those of adenoid cystic carcinoma. Nonetheless, terminal duct carcinoma is cytologically distinctive from adenoid cystic carcinoma and appears to have a more favorable prognosis. PMID- 2990193 TI - Herpes simplex esophagitis--a study in autopsy series. AB - Autopsy cases were reviewed in order to determine the incidence and underlying diseases of herpes esophagitis. Review of both autopsy records and the preserved viscera of 145 cases disclosed 9 cases (6%) of herpes esophagitis. Herpes simplex virus type I specific antigen was detected in all of these nine cases by immunohistochemical method. Eight of nine cases had malignant diseases. Review only of autopsy records of other 254 cases disclosed 5 cases (2%) of herpes esophagitis. Thus, herpes esophagitis is thought to be more common than has previously been appreciated and may be overlooked at autopsy. Although antemortem diagnosis of this condition has been very rare, clinicians should have an accurate knowledge of this disease because herpetic esophageal ulcer may act as a portal of entry for generalized dissemination of virus and other pathogens, and effective antiviral agents are becoming available. PMID- 2990194 TI - Vitamin D-dependent rickets. PMID- 2990195 TI - Chronic syndrome of inappropriate secretion of antidiuretic hormone in childhood. AB - Chronic forms of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) occur infrequently in childhood. We describe three pediatric patients with chronic SIADH. In two patients the persistent SIADH was associated with neoplasms of the central nervous system while in the third patient it was due to congenital anomalies of the central nervous system. We describe the clinical course and management of each patient, including the long-term use of furosemide taken orally once a day. PMID- 2990196 TI - Studies on the mechanism of the lower esophageal sphincter pressure response to alkali ingestion in humans. AB - To clarify the lower esophageal sphincter (LES) pressure response to alkali ingestion, normal subjects and postantrectomy patients with either a gastroduodenostomy or gastrojejunostomy were studied in a double-blind controlled fashion. LES pressure and serum gastrin concentrations were measured after ingestion of a 100 ml bolus of either 0.4 M NaHCO3 or 0.4 M NaCl. In addition, the effect of a therapeutic dose (30 ml) of a commercial antacid preparation was studied in a double-blind fashion in 14 patients with gastroesophageal reflux disease. Peak increases in LES pressure above basal were significantly higher (p less than 0.05) after NaHCO3 than after NaCl in normal subjects and in patients with vagotomy and Billroth I antrectomy, but not in patients with vagotomy and Billroth II antrectomy. Serum gastrin concentrations were unaffected by alkali. Thirty milliliters of liquid antacid containing aluminum and magnesium hydroxide resulted in a small sustained rise in LES pressure over the first 50 min after ingestion, but this was not statistically different than the placebo response. It is suggested that: 1) neither the antrum nor intact vagi nor gastrin were required for NaHCO3 ingestion to increase LES pressure; 2) the increase in LES pressure with NaHCO3 ingestion appears to rely upon an intact duodenum and may relate to volume and osmolarity of the alkali load; and 3) therapeutic doses of a liquid commercial antacid does not significantly increase LES pressure in the presence of an intact stomach. PMID- 2990197 TI - Norwalk virus gastroenteritis. An outbreak associated with a cafeteria at a college. AB - An explosive outbreak of gastrointestinal illness occurred among students and employees at a small college in Florida in November 1980. Common symptoms were diarrhea, nausea, weakness, abdominal cramps, chills, vomiting, and low-grade fever. Cases of illness were identified in 40% of 628 students and 15% of 162 employees who responded to a survey. Among students, there was a sevenfold excess risk associated with eating one or more meals at the campus cafeteria November 3 5 (p much less than 0.001). Tossed salad from one meal was strongly associated with illness (p less than 0.0001). Fecal contamination of the salad was documented, although the source of contamination was not identified. Person-to person spread could not be demonstrated. Seroconversion to Norwalk antigen occurred in significantly more cases (5/6) than noncases (1/6) (p = 0.04). PMID- 2990198 TI - Validation of self-administered questionnaire for study of peripheral neuropathy. AB - An eight-item, self-administered questionnaire for the ascertainment of peripheral neuropathies was validated with 19 persons who had neuropathy confirmed by neurologists' examinations and electrodiagnostic studies, and with 37 persons without known neuropathy. The results from cases and neurology clinic controls suggested a two-stage screening definition for cases of peripheral neuropathy: the classification as cases of all persons who required help getting out of a chair one or more days/week and, among those not requiring help, who had a symptom score of zero or more based on the formula Si = -2.6 + 0.8 (frequency of cramps in arms or legs) + 0.5 (frequency of paresthesias in arms, hands, legs, or feet). This case definition resulted in a sensitivity of 76.5% and a specificity of 93.3% when applied to the original data from which it was developed. The statistical technique of cross-validation suggested an upward bias in these estimates of sensitivity and specificity of 12% and 20%, respectively. The model had a specificity of 100% when tested independently with medical office controls. The authors discuss the use of the questionnaire in cross-sectional epidemiologic studies. PMID- 2990201 TI - Re: Acute hemorrhagic conjunctivitis. PMID- 2990199 TI - Risk factors for gestational trophoblastic disease in Italy. AB - Between June 1981 and March 1983, data were collected to assess risk factors for gestational trophoblastic disease in a case-control study of 100 women with trophoblastic tumors (17 partial hydatidiform moles, 63 complete moles, and 20 choriocarcinomas) and 200 age-matched controls admitted for normal deliveries to university or general hospitals in Lombardy, Northern Italy. Questions were asked about each patient's general life-style, and medical, obstetric, menstrual, contraceptive, and social history. The risk of trophoblastic disease increased with increasing paternal age: women whose husbands were aged 40-44 years and 45 years or more had a relative risk of 2.4 and 4.2, respectively, compared to women married to men aged under 40 years. This association was independent of maternal age. Cigarette smoking was associated with trophoblastic tumors (relative risk estimate for smokers vs. never smokers = 2.0, 95% confidence interval = 1.2-3.2), the risk being greater for women who smoked more cigarettes and for longer. The effect of cigarette smoking was not explained by any other identified potential distorting factor. A positive history of fertility problems or difficulties in conception and a personal or family history of gestational trophoblastic disease were more common among the cases. Past use of oral contraceptives was not related to the risk of trophoblastic tumors, but use of an intrauterine device was significantly more common among the cases. The findings give epidemiologic support to the evidence of an androgenetic role in the origin of hydatidiform mole; moreover, they provide new hypotheses on the risk factors for gestational trophoblastic disease in developed countries. Further exploration of these factors may lead to a more coherent body of evidence on the etiology of these diseases. PMID- 2990200 TI - Risk factors for fatal childhood diarrhea: a case-control study from two remote Panamanian islands. AB - Between September 1979 and March 1980, distinguishing features between fatal and nonfatal cases of diarrhea caused by the same etiologic agents were sought in a case-control investigation of Cuna Indian children living on the San Blas Islands located off Panama's Caribbean coast. The eight fatal cases of diarrhea (four associated with rotavirus, one with Giardia lamblia, and three without identifiable pathogens), which occurred in a cohort of 186 children aged less than five years who were followed for seven months, were matched with 24 contemporaneously occurring nonfatal cases of diarrhea. Weight-for-length measurements falling below the 90th percentile of the reference standard, reliance on traditional rather than equally as available Western medicine, and failure to receive oral rehydration solution were significantly more common among fatal than nonfatal cases. Incorporating traditional medicine men with their long standing village-wide authority into expanded community health education programs that emphasize the importance of early treatment of diarrhea with oral rehydration solution would probably reduce mortality associated with diarrheal illness in this population. PMID- 2990202 TI - Molecular aspects of erythroenzymopathies associated with hereditary hemolytic anemia. AB - Since the discovery of glucose 6-phosphate dehydrogenase (G6PD) and of pyruvate kinase deficiencies, erythroenzymopathies associated with hereditary hemolytic anemia have been extensively investigated. Kinetic and electrophoretic studies have shown that most, if not all, erythroenzymopathies are caused by the production of a mutant enzyme. Except for a few enzymes that are abundant in blood and tissues, it is difficult to obtain enough sample to study the functional and structural abnormalities of mutant enzymes associated with genetic disorders in man. The primary structures of only two normal red cell enzymes which can cause hereditary hemolytic anemia, phosphoglycerate kinase (PGK) and adenylate kinase, have been determined. Single amino acid substitutions of PGK variants have been found, and the identification of the exact molecular abnormalities of such variants has helped us to understand the accompanying functional abnormality. Gene cloning makes possible the identification of the DNA sequence that codes for enzyme proteins. Recently, human complementary DNA (cDNA) for aldolase, PGK, G6PD, and adenosine deaminase (ADA) have been isolated, and the nucleotide sequences for PGK and ADA determined. In the near future, human cDNA sequencing should permit identification of the gene alteration that gives rise to the mutant enzymes. PMID- 2990203 TI - Transfusion-associated AIDS. AB - Acquired Immunodeficiency Disease (AIDS) may be transmitted by transfusion of blood components, although the risk remains extremely small. This report reviews the evidence for transmission by transfusion, the risk in several different situations, and the steps blood banks have taken to prevent transfusion of blood from donors at risk for AIDS. A test for the antibody to the virus responsible for AIDS (HTLV-III) will shortly be in routine use in blood banks, but the frequency with which the results will be falsely negative or falsely positive in a blood donor population remains to be established. PMID- 2990204 TI - Pyramidal tract deficits and polyneuropathy in hyperthyroidism, Combination clinically mimicking amyotrophic lateral sclerosis. AB - Generalized weakness, intermittent dysphagia, and a 40-pound weight loss developed in an elderly man over a six-month period. Examination revealed weakness, atrophy and fasciculations of extremity musculature, pseudobulbar speech, hyperactive upper extremity reflexes, and extensor toe signs without sensory loss. Results of electrodiagnostic studies were consistent with an axonal polyneuropathy. Endocrinologic results were compatible with hyperthyroidism. Radioiodine therapy resulted in resolution of clinical neurologic symptoms and signs within seven months. This case illustrates a previously undescribed concurrence of hyperthyroid associated polyneuropathy and pyramidal tract dysfunction that led to an initial clinical diagnosis of amyotrophic lateral sclerosis. PMID- 2990205 TI - Analysis of T cell subsets in different clinical subgroups of patients with the acquired immune deficiency syndrome. Comparison with the "classic" form of Kaposi's sarcoma. AB - Ninety patients were grouped according to three different forms of acquired immune deficiency syndrome (AIDS): Kaposi's sarcoma "recent outbreak" type (38), reactive lymphadenopathy (27), and opportunistic infections (17), and a fourth group of patients with "classic" Kaposi's sarcoma (8). All patients with "classic" Kaposi's sarcoma were previously treated with electron-beam irradiation. These four groups were compared with 40 normal control subjects. Peripheral blood mononuclear cells isolated by density separation were reacted with a panel of mouse monoclonal antibodies that recognizes all peripheral blood T cells (OKT3-positive), helper (OKT4-positive), and suppressor (OKT8-positive) T cell subsets. The OKT4/OKT8 ratio was used to define the balance between these two subsets. All three groups with AIDS with or without Kaposi's sarcoma showed a decrease in the OKT4/OKT8 ratio. The group with "classic" Kaposi's sarcoma showed individual T cell subset values that were also abnormal. These findings confirm the previously reported imbalance of T cell subsets in patients with AIDS and Kaposi's sarcoma, which is also evident in patients with treated "classic" Kaposi's sarcoma. PMID- 2990206 TI - Diagnostic strategies in osteomyelitis. AB - Technetium-99 (99Tc) pyrophosphate bone scanning often identifies patients with osteomyelitis before roentgenographic findings appear. However, recent studies have shown that 99Tc bone scanning often gives false-negative results, especially in neonates. The accuracy of computed tomographic scanning and indium-111 leukocyte scanning for diagnosis of early osteomyelitis has not been established. 99Tc bone scanning often gives false-positive results in patients with other conditions leading to bone injury and repair, such as trauma or recent surgery, further limiting the usefulness of this imaging procedure. Newer imaging techniques have not been adequately evaluated to establish their specificity. Because of their high cost and unproved accuracy, these new imaging procedures should not be routinely applied until their usefulness has been established. Bone biopsy remains the procedure of choice for establishing the diagnosis in patients suspected clinically to have osteomyelitis with negative findings on roentgenography and 99Tc bone scanning. Although Staphylococcus aureus is the leading cause of osteomyelitis, other pathogens cause 30 to 40 percent of cases. Aspiration or biopsy of the involved bone is usually required to choose appropriate antibiotic therapy. Bone biopsy is essential in chronic osteomyelitis, since cultures of sinus drainage are unreliable. Osteomyelitis in diabetics with foot infection and in association with decubitus ulcers presents special problems. Radionuclide scanning often give false-positive results in these patients. Proper diagnosis usually requires careful assessment of clinical and roentgenographic findings. Although bone biopsy seems useful in diagnosing osteomyelitis underlying decubitus ulcers, its role in diabetic patients with foot infections is not established. I do not recommend biopsy of foot bones in diabetic patients, since culture of bone biopsy specimens often give unreliable results in these situations because of contamination with ulcer organisms. PMID- 2990208 TI - Bruising and thin skin in a 54-year-old woman. PMID- 2990207 TI - Prevention and treatment of infectious diarrhea. Speculations on the next 10 years. AB - The next decade should explode with exciting schemes and novel agents for the prevention and treatment of infectious diarrhea. The development of oral, nonabsorbed antibiotics will continue, but new antidiarrheal drugs, such as gastrointestinal hormone analogues and alpha-adrenergic agonists, will be added to our therapeutic armamentarium. Improved oral rehydration solutions, such as glycine in electrolyte solution, promise to revolutionize the management of diarrhea by diminishing diarrheal stool volume to the point where losses are too small to be clinically relevant. Infant formulas and adult oral solutions fortified with antibodies raised against selected enteropathogens may provide a way to prevent infectious diarrheas in infants and travelers. Advances in genetic engineering will usher in a new era of experimental and licensed enteric vaccines, including those against cholera, Escherichia coli, Shigella, typhoid fever, and rotavirus. PMID- 2990209 TI - Long-term survivors of small cell carcinoma of the lung. AB - Small cell carcinoma of the lung has been shown to be exquisitely responsive to chemotherapy. Unfortunately, these responses are often short in duration and long term disease-free survival is infrequent. This review of the records of all patients with small cell carcinoma of the lung treated on protocol at The Johns Hopkins Oncology Center from 1973 to 1982 showed that 25 of 225 (11.1 percent) survived two years or longer. Patients with limited disease (20 of 94) and patients with a complete response (15 of 72) had greater two-year survival than those with extensive disease (five of 131) or partial remission (eight of 104). However, 18 of the 25 long-term survivors eventually had relapses, and relapse occurred as late as eight years after diagnosis. This study further emphasizes the impressive discrepancy between the rate and magnitude of the initial response and ultimate survival in patients with small cell carcinoma of the lung. PMID- 2990211 TI - TORCH tests and what they mean. AB - TORCH tests can be used to document immunity or recent infections. Many laboratories now offer TORCH tests and there are a large variety of methods, kits, and reporting systems. Some of the TORCH tests are not accurate and should be avoided. In general, we prefer enzyme immunoassays for documentation of past infections or immunity. To demonstrate recent infections with toxoplasmosis and rubella we use IgM enzyme-linked immunosorbent assay tests but for cytomegalovirus and herpes infections we prefer virus isolation. Physicians should send specimens only to laboratories that participate in proficiency surveys or similar tests that document the accuracy of the laboratory. All TORCH test sera should be held by the laboratory for 1 year to permit repeat testing and analysis with subsequent specimens if necessary. PMID- 2990210 TI - Neuroendocrinology of opioid peptides and their role in the control of gonadotropin and prolactin secretion. AB - Substantial evidence now exists to indicate that the endogenous hypothalamic opioidergic mechanism(s) represents one of the important controlling systems for release of gonadotropin-releasing hormone. Modulations of frequency and amplitude of the secretory activity of gonadotropin-releasing hormone appears to be mediated through an inhibitory action of endogenous opioids, and the functional coupling of the opioidergic and gonadotropin-releasing hormone systems is an ovarian steroid-dependent event. There is also evidence to implicate suprahypothalamic mechanism(s) that enhance endogenous opioid inhibition of secretion of gonadotropin-releasing hormone. Although exogenous opioid peptides and their synthetic analogs consistently induce the secretion of prolactin, blockade of opioid receptors in humans by naloxone failed to elicit a decrement in the levels of prolactin under a variety of conditions. On the contrary, naloxone induced a remarkable increment in the secretion of prolactin via an increased frequency of pulsatile release which is synchronized with pulses of luteinizing hormone. These observations suggest that a common neuroendocrine mechanism is involved in the opioidergic control of the secretion of both luteinizing hormone and prolactin in women. PMID- 2990212 TI - Superficial laser vulvectomy. III. A new surgical technique for appendage conserving ablation of refractory condylomas and vulvar intraepithelial neoplasia. AB - Despite the unique properties of the carbon dioxide laser, many surgeons do not know how to exploit the full potential of this sophisticated instrument. Effective laser operation on the vulva depends upon the accuracy of delineation of disease, the use of optimum power densities, and the ability to exercise precise control over depth of ablation. This article describes a surgical technique that capitalizes upon these principles, thereby maximizing the margin between favorable and poor outcomes. Superficial laser vulvectomy is a safe and efficient procedure in the hands of expert physicians, but should not be attempted by those who are less experienced. Indications for this operation and safeguards against surgical misadventure are also discussed. PMID- 2990213 TI - Effect of the herpes simplex virus genome on the response of infection to corticosteroids. AB - The type and severity of ocular herpetic disease, as well as the pattern of recurrence, have been shown to be determined by the virus genome. We infected rabbit eyes with two closely related recombinant strains of herpes simplex virus type 1 and treated one half of the eyes in each group with corticosteroids before or immediately after virus inoculation. The severity of disease in the first week was similar in the treated and untreated eyes infected with the F(MP)F strain; however, with F(MP)E infection, the disease in the treated eyes was significantly worse than the disease in the untreated eyes. Cultures of corneal virus showed similar titers in all of the groups, but cultures of trigeminal ganglia indicated that increased severity of disease did not result in an increased tendency toward ganglionic colonization. The results suggest that the response to corticosteroids is another factor that is determined by the genetics of the infecting virus, but that there is no correlation between worsening of disease with corticosteroid treatment and the establishment of virus latency. PMID- 2990214 TI - Regulation of aqueous flow by the adenylate cyclase receptor complex in the ciliary epithelium. AB - The answer to how the beta-adrenergic receptor mediates a fall in intraocular pressure has been elusive. Methods of measurement have not been refined sufficiently. The separate changes after adrenergic treatment frequently are small, and the tissue effects are multiple. On a molecular basis, stimulation of the beta-adrenergic receptor activates intracellular adenylate cyclase to produce increased cyclic adenosine monophosphate. Acting by different cell-receptor mechanisms, but nonetheless potent, nonadrenergic stimulators of adenylate cyclase in the ciliary epithelium, such as cholera toxin and organic fluorides, have been studied in experimental animals. They reduce intraocular pressure by reducing net aqueous flow. When forskolin, a diterpene and potent stimulator of adenylate cyclase, became available, it was used in noninvasive topical form in the human eye to clarify the question of whether increased cyclic adenosine monophosphate reduces intraocular pressure and aqueous flow. Noninvasive studies in human eyes have demonstrated a 35% reduction in outflow pressure after the administration of forskolin in a 1% topical suspension, matched by a corresponding reduction in aqueous flow. Tonographic outflow facility was unaltered. Thus, the entire reduction in intraocular pressure can be accounted for by a reduction in net aqueous flow. PMID- 2990215 TI - Methyl bromide optic atrophy. AB - A 32-year-old fumigation assistant developed systemic and neuro-ophthalmic manifestations of methyl bromide poisoning, including increased serum bromide level (6.6 mg/100 ml), paresthesias and burning dysesthesia on his hands and feet, and visual impairment. Ocular examination showed mild bilateral decrease in vision, temporal optic nerve head pallor, severely attenuated visual-evoked response amplitudes and normal latencies, a normal electroretinogram, an abnormal electrooculogram, and a severe deuteranomalous (green) defect on Farnsworth Munsell 100-hue testing. His vision had not improved 12 months after the initial exposure. PMID- 2990216 TI - Effect of the sympathetic nervous system on accommodation. AB - The accommodative effects of alpha- and beta-adrenergic stimulation were investigated in human subjects by measuring changes in the response ACA ratio and in the response accommodative amplitude as a result of instilling hydroxyamphetamine hydrobromide and phenylephrine hydrochloride. Statistically significant increases in the ACA ratios were found to result from hydroxyamphetamine hydrobromide, but not from phenylephrine hydrochloride. The accommodative amplitude significantly decreased under the influence of each agent. It was concluded that these adrenergic effects on accommodation were mediated through the beta-receptor sites. This provides further evidence of the dual autonomic innervation of accommodation. PMID- 2990217 TI - Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. AB - A series of human carcinoma cell lines was examined for human papillomavirus (HPV) DNA sequences with the use of HPV-6, HPV-11, HPV-16, and HPV-18 DNA probes. Six of eight cell lines derived from human cervical carcinomas were shown to contain integrated HPV DNA sequences. In five of these six lines, HPV-specific polyadenylated RNA species could also be identified. The expression of HPV sequences was detected in three lines with a HPV-18 DNA probe and in two lines with a HPV-16 DNA probe. Of the two lines which contained HPV-16 specific RNA, one contained HPV DNA sequences which hybridized only to an HPV-16 probe, and the other contained HPV DNA sequences which hybridized to both HPV-16 and HPV-18 DNA probes. Six cell lines established from human squamous-cell carcinomas of the bladder, pharynx, lung, esophagus, and vulva were negative for HPV-6, HPV-11, HPV 16, and HPV-18 DNA sequences under stringent hybridization conditions. PMID- 2990218 TI - Lipid peroxidation and acute lung injury after thermal trauma to skin. Evidence of a role for hydroxyl radical. AB - The authors have previously shown that thermal injury to the skin of rats results in the development of acute lung injury that is susceptible to systemic treatment of animals with catalase and dependent on the presence of neutrophils. The current studies have been expanded for exploration of the nature of the neutrophil-derived oxygen products responsible for the lung injury and have also focused on evidence of the appearance of products of lipid peroxidation (conjugated dienes). With respect to the former, treatment of rats with iron chelators (deferoxamine mesylate, 2,3-dihydroxybenzoic acid), with scavengers of hydroxyl radical (dimethyl sulfoxide, dimethyl thiourea, sodium benzoate), or with vitamin E affords a significant degree of protection from acute lung injury as assessed by changes in lung vascular permeability and by morphologic parameters. These data suggest that lung vascular injury after thermal trauma of the skin is related to the generation by neutrophils of the hydroxyl radical. Conjugated dienes have been demonstrated to appear sequentially both in the burned skin (at 1/4 hour) and in the lungs (at 2 hours), as well as in the plasma (with peaks at 1/2 and at 3 hours) after thermal injury. The appearance of the conjugated dienes in plasma at the two intervals of time is greatly diminished if animals are pretreated with the iron chelator deferoxamine, with catalase, or with scavengers of hydroxyl radical. Furthermore, the appearance of conjugated dienes in plasma at 30 minutes and 3 hours is significantly diminished if animals are depleted of neutrophils, complement-depleted, or the burned skin is excised immediately after thermal injury. These data indicate a linkage between thermal trauma of skin, secondary injury of lung, and appearance in plasma and tissues of products of lipid peroxidation. PMID- 2990220 TI - Immunoelectron-microscopic localization of Tac antigen in adult T-cell leukemia/lymphoma. AB - The localization of Tac antigen in adult T-cell leukemia-associated antigen (ATLA)-positive lymphomas was studied ultrastructurally with the use of the immunoperoxidase technique. The antigen was observed on the plasma membranes of a portion of the characteristic cells with convoluted nuclei and a majority of the cells with less irregular nuclei, which were larger than the former. In addition, the cisternae of the rough endoplasmic reticulum, perinuclear cisternae, and Golgi cisternae of the latter cells were also positively stained with anti-Tac antibody. It is thought that the positive reaction of the plasma membranes may correspond to interleukin 2 (IL 2) receptors, the cytoplasmic and perinuclear reaction sites may well correspond to the precursor of IL 2 receptors, and Tac antigen may be produced in the cytoplasm of the ATLA-positive lymphoma cells. PMID- 2990219 TI - Bronchiolitis obliterans and pneumonia induced in young dogs by experimental adenovirus infection. AB - Young beagle dogs were experimentally inoculated with canine adenovirus Type 2 and studied by virologic, histologic, immunoperoxidase, and ultrastructural methods from 1 to 26 days after inoculation. Virus was recovered from lungs at 2, 3, 5, and 8 days after inoculation. Virions and viral antigen were demonstrated by ultrastructural and immunoperoxidase techniques in nonciliated bronchiolar epithelial cells and mucous cells in bronchioles, bronchi, and trachea as well as in bronchial and tracheal submucosal gland epithelial cells. Viral replication in airways was associated with a severe necrotizing and proliferative bronchitis and bronchiolitis. Virus and viral antigen were demonstrated in Type 2 alveolar epithelial cells and were associated with interstitial pneumonia. Partial and complete stenosis of bronchioles by connective tissue was observed at 15 and 26 days after inoculation. There was a 50% reduction (P less than 0.02) in mean terminal bronchiolar cross-sectional area in the right middle lobe of virus infected dogs at 26 days after inoculation. It is concluded that experimental adenovirus infection in dogs induces bronchiolitis obliterans and that this experimental model may be useful for studies on adenovirus-induced lung injury during early life. PMID- 2990222 TI - Malignant rabbit fibroma syndrome. A possible model for acquired immunodeficiency syndrome (AIDS). PMID- 2990221 TI - Activated rat neutrophils. Correlation of arachidonate products with enzyme secretion but not with O(2)- generation. AB - Functional responses (enzyme secretion and generation of O2-) have been studied in rat neutrophils with the use of a variety of different agonists which vary in their ability to activate neutrophils. Concomitantly, the authors have analyzed the cells for production of cyclooxygenase (PGE2, PGF2 alpha, and TXB2) and lipoxygenase (monoHETEs and LTB4) products in order to determine whether there is a correlation between functional responses and arachidonate products in stimulated neutrophils. The studies indicate that enzyme secretion is closely correlated with generation of arachidonate (cyclooxygenase as well as lipoxygenase) products. These responses are dependent on the dose of agonist employed, but there does not appear to be a unique pattern of arachidonate products that can be attributed to a specific agonist. With respect to monohydroxyeicosatetraenoic acid (monoHETE) generation, we demonstrate that stimulated rat neutrophils selectively produce 5-HETE, to the virtual exclusion of 12-HETE and 15-HETE. The production of O2- from stimulated rat neutrophils is correlated neither with enzyme secretion nor with the generation of arachidonate products. These studies suggest that enzyme secretion and O2- generation are mediated by two fundamentally different intracellular pathways, even though these functional responses probably derive from common agonist-receptor interactions on the plasma membrane. PMID- 2990223 TI - The role of neutrophils in the development of cadmium chloride-induced emphysema in lathyrogen-fed hamsters. AB - This study was designed to evaluate the role of neutrophils (PMNs) in the pathogenesis of emphysema. After administration of CdCl2 intratracheally to hamsters fed a lathyrogen, beta-amino propionitrile fumarate (BAPN), the classic lesions of emphysema developed. Administration of specific antineutrophil serum markedly reduced the PMN influx into the lungs which followed CdCl2 exposure and produced a substantial decrease in elastase and collagenase content of bronchoalveolar lavage fluid (BAL). The degree of emphysema in PMN-depleted hamsters given BAPN and CdCl2 was not different from that in hamsters given BAPN and CdCl2. Furthermore, the degree of acute lung injury following CdCl2, as determined by hydroxyproline content of BAL and by lung lipid peroxidation, was the same in PMN-depleted and non-depleted groups. Thus, PMNs were not necessary for the induction of emphysema in BAPN-CdCl2-treated hamsters. The results suggest that PMNs may not be obligate mediators of emphysema. PMID- 2990224 TI - Intracellular pH in human fibroblasts: effect of mitogens, A23187, and phospholipase activation. AB - Serum-deprived human fibroblasts (HSWP cells) were loaded with either the fluorescent pH indicator 6-carboxy-4',5'-dimethylfluorescein or the calcium indicator quin 2, and the fluorescence of the intracellular probes was continuously monitored with a microspectrofluorometer. Addition of a cocktail of peptide growth factors causes intracellular alkalinization, which is blocked by amiloride or by replacement of extracellular Na+ with choline, confirming that mitogenic stimulation activates a Na+-H+ exchanger in HSWP cells. The exchanger is also activated by A23187, acid loading the cells, and stimulation of phospholipase activity with melittin. Growth factors and melittin activation of the exchanger have been demonstrated to be dependent on the mobilization of Ca2+ from intracellular stores. The intracellular alkalization and increase in Ca2+ activity due to both melittin and growth factors is inhibited by phospholipase inhibitors, indicating that phospholipase activity is necessary for the activation of the Na+-H+ exchanger and the mobilization of intracellular Ca2+. PMID- 2990225 TI - Baboon erythrocyte ghosts contain beta-adrenergic receptors. AB - We have used the beta-adrenergic antagonist [3H]dihydroalprenolol [( 3H]DHA) to identify binding sites on the erythrocyte membrane of the primate Papio ursinus. Analysis of the saturation isotherm revealed binding to be saturable with a maximal number of binding sites of 499 fmol/mg protein. [3H]DHA binds specifically to the erythrocyte ghosts with an apparent dissociation constant (Kd) of 0.57 +/- 0.06 nM. A similar value for Kd (0.46 +/- 0.07 nM) was evaluated from the rate constants of association (0.013 +/- 0.003 X nM-1 X min-1) and dissociation (0.006 +/- 0.001 X min-1). beta-adrenergic agonists compete for the binding sites with an order of potency (dl-isoproterenol greater than l epinephrine greater than l-norepinephrine) typical of a beta 2-adrenergic receptor. Binding was shown to be stereospecific with l-stereoisomers being more potent than their corresponding d-stereoisomers in causing half-maximal inhibition. Isoproterenol stimulated the production of intracellular adenosine 3',5'-cyclic monophosphate (cAMP) in a concentration-dependent manner, maximal levels (1.130 +/- 0.358 pmol cAMP/10(8) cells) being four times the basal levels. The results demonstrate the existence of a large number of beta-adrenergic receptors on baboon erythrocyte ghosts. PMID- 2990226 TI - A Na+-Ca2+ exchange process in isolated sarcolemmal membranes of mesenteric arteries from WKY and SHR rats. AB - The existence of a Na+-Ca2+ exchange process in cell membrane vesicles isolated from mesenteric arteries of Wistar-Kyoto normotensive (WKY) and spontaneously hypertensive (SHR) rats was investigated. Membranes from cleaned mesenteric arteries were isolated by sucrose density gradient centrifugation, which yielded three distinct membrane fractions. The lighter membrane fraction of both WKY and SHR rats was enriched in 5'-nucleotidase activity, a marker for cell membrane, by about 10-fold, based on the activity in the homogenate, and was higher in membranes of SHR compared with WKY rats. Ouabain-sensitive Na+-K+-ATPase activity, another marker for cell membrane, was also concentrated in the lighter membrane fraction and was lower in the membranes of SHR compared with WKY rats. Higher activities of 5'-nucleotidase and Na+-K+-ATPase of both WKY and SHR rats was taken as evidence that the lighter membrane fraction was enriched in plasma membrane. Electron microscopic examination indicated that the membranes were in vesicular form. When the vesicles were loaded with Na+, a time-dependent uptake of Ca2+ was observed if the assay was carried out in high potassium to create a Na+ concentration gradient across the membrane of the vesicles. Very little Ca2+ uptake was observed when the vesicles were loaded with K+ or when the uptake of Ca2+ was carried out under conditions in which the Na+ gradient across the vesicle membranes was reduced. Ca2+ uptake in Na+-loaded vesicles of SHR rats was only slightly increased compared with WKY rats. The data indicate that a Na+-Ca2+ exchange process exists in the cell membrane of rat mesenteric arteries. PMID- 2990227 TI - Role of prostaglandins in hypoxia-stimulated erythropoietin production. AB - The role of prostaglandins in the mediation of hypoxia-stimulated erythropoietin (Ep) production by cultured rat renal mesangial cells was examined. It was found that an increase in prostaglandin E2 (PGE2) production accompanied the rise in Ep due to hypoxia (2% O2). The hypoxia-stimulated increase in Ep production was abolished in the presence of the cyclooxygenase inhibitor indomethacin (10(-5) M). When PGE2 (10(-6) M was added simultaneously with indomethacin, however, no diminution in hypoxia-stimulated Ep production was observed. Addition of arachidonic acid (AA, 10(-5) M), PGE2 (10(-6) M), or PGI2 (10(-4) M) enhanced Ep production under normoxic conditions (20% O2), while PGF2 alpha (10(-6) M) had no effect on Ep production. AA, PGE2, and PGI2 were found to stimulate adenosine 3',5'-cyclic monophosphate formation by the cultured mesangial cells. Enhancement of adenylate cyclase activity by forskolin (10(-5) M) also increased Ep production in the cell cultures. Our results suggest that hypoxia-stimulated Ep production by cultured mesangial cells is mediated by prostaglandins with subsequent stimulation of adenylate cyclase activity. PMID- 2990228 TI - Agonist-induced isotonic contraction of cultured mesangial cells after multiple passage. AB - Contractile cells under conditions of prolonged culture lose their ability to contract in the usual manner (i.e., isotonically). One explanation for this may be that contraction is prevented by tight cell-to-substrate adhesion. Two models in which substrate adhesiveness was expected to be diminished were used to test this hypothesis. In one, cells were seeded onto collagen-coated dishes and used within 40 min of plating. In the other, cells were plated onto dishes coated with poly-2-hydroxyethyl methacrylate (poly-HEMA) and used, depending on thickness of the poly-HEMA substrate, up to periods of 1 wk. Cells plated onto such substrates contracted when challenged with either PGE2 (2 X 10(-6) and 2 X 10(-9) M), arginine vasopressin (AVP, 10(-6)-10(-9) ), or the calcium ionophore A23187 (5 micrograms/ml). Contraction took place within 5-15 min at 37 degrees C. The contraction seen with AVP was due to its pressor action because 1-desamino-8-D arginine vasopressin (dDAVP), the antidiuretic analogue, did not cause contraction and the anti-pressor analogue [1-(beta-mercapto-beta beta cyclopentamethylene propionic acid)-4-valine 8-D-arginine]-vasopressin [d(CH2)5 VDAVP] blocked contraction by AVP. The contraction seen with AVP was dependent on extracellular calcium, whereas that observed with prostaglandin E2 (PGE2) was not.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990229 TI - Very small fat cell populations determined by a modified osmium tetroxide-urea method. AB - To facilitate investigations on very small fat cell (VSFC) populations in adipose tissue, an alternate method of preparing fat tissue samples was explored. The osmium tetroxide-8M urea method, modified by addition of a 95% ethanol step in tissue processing, centrifugation between steps, and final resuspension in 55% glycerol in 0.01% Triton-saline, was compared with the collagenase method for determination of VSFC populations in Fischer 344 epididymal and Sprague-Dawley retroperitoneal adipose depots. For each method and in both depots, the average histogram of 300 adipocyte diameters, measured by microscopy, was bimodal with the nadir between 30 and 40 micron diameter. The average histogram of fat cells less than 35 micron in diameter showed a separate population of VSFC existed in each depot. The modified osmium-urea method gave better results and was easier to perform than the collagenase method. It has confirmed our earlier results and raises anew questions concerning a role for the natural existence of a VSFC population in the adipose depot. PMID- 2990230 TI - Binding protein for vitamin D and its metabolites in rat mesenteric lymph. AB - A protein with high affinity for vitamin D3 and 25-hydroxyvitamin D3 in rat mesenteric lymph has been studied. Mesenteric lymph was collected after duodenal instillation of radiolabeled vitamin D3 and 25-hydroxyvitamin D3. As previously described, approximately 10% of vitamin D3 (S. Dueland, J.I. Pedersen, P. Helgerud, and C.A. Drevon, J. Biol. Chem. 257: 146-150, 1982) and 95% of 25 hydroxyvitamin D3 (S. Dueland, J.I. Pedersen, P. Helgerud, and C.A. Drevon, Am. J. Physiol. 245 (Endocrinol. Metab. 8): E463-E467, 1983) recovered in mesenteric lymph were associated with the alpha-globulin fractions. The radioactive vitamin D3 recovered in the lymph fraction with d greater than 1.006 (free of chylomicrons) coeluted with purified rat serum binding protein for vitamin D and its metabolites (DBP) from an antirat DBP column. The results obtained by immunoblotting after sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that this protein in mesenteric lymph had molecular weight and immunological properties identical with purified serum DBP. Purified serum DBP labeled with 125I was injected intravenously and mesenteric lymph was collected. in lymph, suggesting that DBP may be transferred from blood to mesenteric lymph and that plasma and lymph DBP may have a similar origin. PMID- 2990231 TI - Somatomedin C augments FSH-induced differentiation of cultured rat granulosa cells. AB - Growth hormone (GH) deficiency in rats is associated with decreased ovarian steroidal responsiveness to gonadotropins, possibly through a reduction in the production of the GH-dependent Somatomedin C/insulinlike growth factor I (SM C/IGF I). We have investigated the direct effects of synthetic SM C/IGF I on gonadotropin-stimulated ovarian steroidogenesis in vitro. Granulosa cells were cultured in a serum-free medium for 48 h in the presence of follicle-stimulating hormone (FSH), with or without SM C/IGF I. FSH dose-dependently increased both estrogen and progestin production. Concomitant treatment with SM C/IGF I led to a dose-dependent augmentation of progestin secretion over the full range of FSH doses tested, by a maximum of 2.3- to 2.6-fold. FSH-stimulated estrogen was enhanced by up to 2.4-fold but only at low doses of FSH. SM C/IGF I-enhanced progestin production was associated with increased pregnenolone production and 3 beta-hydroxysteroid dehydrogenase activity, whereas augmented estrogen production appeared to be due to enhanced aromatase activity. The actions of SM C/IGF I, at physiologically relevant concentrations were correlated with increased extracellular cAMP accumulation and cellular protein content but were independent of any change in cell number or viability. In contrast to SM C/IGF I, the closely related peptide multiplication-stimulating activity decreased estrogen production while increasing progestin metabolite accumulation. The present results indicate that the GH-dependent peptide SM C/IGF I may play a role in ovarian development by enhancing gonadotropin-stimulated granulosa cell steroidogenesis. PMID- 2990232 TI - Effects of hypophysectomy and GH administration on bovine and human GH binding to rat liver membranes. AB - Experiments were conducted to investigate the specific binding of highly purified bovine and human growth hormones (bGH and hGH) to purified liver plasma membranes of male rats at various times after hypophysectomy and after the acute intravenous administration of bGH. Liver membranes prepared from hypophysectomized male rats showed a two- to threefold increase in the specific binding of either [125I]iodo-bGH or [125I]iodo-hGH, when compared with membranes prepared from the livers of age-matched normal male rats. The increase in GH binding was apparent within 3 days after hypophysectomy and persisted for a number of weeks after the operation. The increase in GH binding produced by hypophysectomy appeared to be due to an increase in the number of binding sites present on the membranes. The intravenous injection of 200 micrograms of bGH into hypophysectomized male rats 5-60 min before they were killed markedly reduced the ability of liver membranes prepared from these animals to bind [125I]iodo-bGH specifically. This decrease in GH binding seen after the injection of bGH may have been due to the development of a slowly dissociating hormone-binding site complex, which thereby reduced the number of available binding sites. This conclusion is supported by the finding that bGH, which is bound in vitro to isolated liver membranes, dissociates slowly and incompletely in the presence of an excess of unlabeled hormone. Moreover, the degree to which the bound hormone can dissociate appears to depend on the length of time that association is allowed to occur.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990233 TI - Time course of changes in gluconeogenic enzyme activities during exercise and recovery. AB - Gluconeogenic enzymes were assayed after varying periods of exercise and recovery to determine how rapidly changes occur and whether they persist after the cessation of exercise. Untrained male rats (250 g) ran on a treadmill at 28 m/min and were killed after varying periods of exercise and recovery. Livers were quickly removed and analyzed for maximal enzyme activities (saturating levels of substrate) and submaximal activities (low-substrate concentrations). The most significant enzyme changes during exercise were increased maximal activity of phosphoenolpyruvate carboxykinase (PEPCK) and decreased submaximal activity of phosphofructokinase (PFK). Submaximal PFK activity was decreased by 30 min of exercise and remained at that low level up to exhaustion (172 +/- 16 min). Changes in submaximal PFK activity are in response to decreased concentrations of fructose-2,6-bisphosphate that were decreased to approximately one-tenth the control value after 30 min of exercise and remained low throughout exercise and 1 h of recovery. The PEPCK activity progressively increased during exercise and was highest at exhaustion. The cAMP level was significantly elevated in liver of rats exercised for 30 min and continued to rise with duration. Six hours after exercise PEPCK and submaximal PFK activities were the same in control and exercised-rested rats. The change in PEPCK activity is consistent with an increase in the rate of enzyme synthesis and/or a decrease in enzyme degradation during exercise, whereas the lowered activity of PFK likely reflects covalent modification of 6-phospho-fructo-2-kinase/fructose-2,6-bisphosphatase. PMID- 2990234 TI - Metabolic adaptation to reduced muscle blood flow. I. Enzyme and metabolite alterations. AB - A rat model was developed in which the adaptive effects of exposing skeletal muscle tissue to a reduced blood flow during muscle contractions could be studied. The common iliac artery was ligated in one hindlimb, using the other as control. This procedure reduced the exercise blood flow to the individual muscles of the lower limb by 76-93%, evaluated with the microsphere technique. Muscle contractions were induced by electrical stimulation of the sciatic nerves in both legs. After intermittent stimulation for 6 days, a significant increase in citrate synthase and cytochrome c oxidase activities was found in the soleus (26%) and extensor digitorum longus (EDL, 20%) muscles of the ligated legs compared with the control legs. Resting metabolite concentrations were also measured, and a reduction of the ATP level (soleus 35%, EDL 14%) and an increased glycogen content (55-71%) were found. These results demonstrate that a reduced blood flow during muscle contractions provokes an adaptive increase of the oxidative enzyme capacity as well as altered resting levels of intracellular metabolites. PMID- 2990235 TI - Coupled Na+-H+ exchange in isolated acinar cells from rat exocrine pancreas. AB - Isolated acinar cells from the rat exocrine pancreas were loaded with 6 carboxyfluorescein diacetate (CFDA), and the intracellular pH (pHi) was estimated from the pH-dependent fluorescence intensity of trapped 6-carboxyfluorescein liberated from CFDA by intracellular esterases. The intracellular fluorescence intensity was calibrated by equilibrating the internal and external pH with nigericin in K+ buffers. In the absence of Na+ (130 mmol/l K+) a pHi of 6.86 +/- 0.04 was found; in its presence (130 mmol/l Na+) a pHi of 7.17. Acute addition of Na+ increased intracellular pH with increasing Na+ concentrations, reaching a maximum at 150 mmol/l with an apparent Km of approximately 40 mmol/l. Of the different cations tested on pHi, such as Li+, K+, Rb+, and Cs+, only Li+ showed an effect on pHi similar to that of Na+. Amiloride dose dependently inhibited both Na+- and Li+-induced alkalinization (apparent Km approximately 10(-5) mol/l). In the presence of ouabain pHi was decreased by 0.2 pH units. Intracellular acidification induced by permeable buffers such as acetic acid acetate or CO2-HCO3- was dissipated more rapidly in the presence of Na+ compared with K+ or with Na+ and amiloride in the medium. In Li+-preincubated cells intracellular acidification was higher in the absence of Li+ in the extracellular medium than in its presence. This Li+ gradient-induced acidification was dependent on the extracellular pH, was highest at an extracellular pH of 7.05, and decreased with increasing pH to 7.5. The results allow the conclusion that a coupled Na+-H+ exchange is present in pancreatic acinar cells and that the intracellular pH rather than the extracellular Na+ concentration regulates this transport mechanism. PMID- 2990236 TI - Development of gastrointestinal mucosal barrier. VII. In utero maturation of microvillus surface by cortisone. AB - When studying mucosal barrier function of developing animals, we noted that intestinal microvillus membranes (MVM) of newborn animals differ in their fluidity and binding characteristics to lectins compared with adult MVM. To further investigate these differences and determine whether maturation of the microvillus surface could be accelerated in utero, pregnant rats were given intraperitoneal cortisone beginning on the 17th day of gestation. Control and cortisone-treated animals were allowed to deliver normally, and the small intestines from newborns were used to isolate MVM. Microvillus membrane surface characteristics were evaluated by employing an 125I-labeled fucose-specific lectin, Ulex europeus (UEA). Changes in MVM proteins were monitored by disaccharidase activities and sodium dodecyl sulfate-polyacrylamide electrophoresis. MVM fluidity was accessed using a 5-doxyl stearic acid label and electron-spin-resonance spectroscopy. Results from these studies indicate that the birth weights of newborn rats exposed to cortisone in utero were significantly reduced; sucrase activity was prematurely induced and specific activities of lactase and maltase were enhanced in the intestines of the cortisone-treated newborns as contrasted with control animals. Furthermore, binding of 125I-UEA to MVM was greatly increased in treated animals. MVM fluidity decreased (P less than 0.001) compared with control animals and resembled the structural characteristics of more mature MVM. These results suggest that cortisone exposure in utero accelerate maturation of the microvillus surface of enterocytes. PMID- 2990237 TI - Juxtamedullary nephrons during acute metabolic alkalosis in the rat. AB - The renal handling of bicarbonate during acute metabolic alkalosis was examined in Munich-Wistar rats using micropuncture techniques. Group I received an acute bicarbonate load, and fractional delivery of total CO2 (tCO2) (FDtCO2) to the superficial late distal tubule (LD) was significantly lower than to the base of the papillary collecting duct (B) (18.4 +/- 1.7 vs. 22.9 +/- 1.5%; P less than 0.01), indicating net addition of bicarbonate between LD and B. When acutely bicarbonate-loaded rats had their deep nephrons destroyed with bromoethylamine hydrobromide (BEA) (group II), net addition of tCO2 between LD and B was abolished and net reabsorption uncovered (FDtCO2 LD: 28.0 +/- 3.6 vs. B: 17.5 +/- 2.5%; P less than 0.01). The infusion of amiloride (2.5 mg/kg body wt) to alkalotic rats treated with BEA (group III) completely inhibited distal bicarbonate reabsorption but did not reestablish addition (FDtCO2 LD: 27.6 +/- 1.6 vs. B: 26.1 +/- 3.7%; P = NS). The values obtained for sham-operated animals (group IV) were the same for group I. The patterns that were observed between LD and B were reproduced for the four groups of animals when FDtCO2 LD was compared with the fractional excretion of bicarbonate in the urine of the intact contralateral kidney. These studies suggest that juxtamedullary nephrons contribute a higher load of bicarbonate than superficial nephrons to the final urine during acute metabolic alkalosis in the rat. PMID- 2990238 TI - Effects of vasopressin on phosphoinositides and prostaglandin production in cultured mesangial cells. AB - We studied the effects of vasopressin on phospholipid metabolism in mesangial cells and the temporal relationship of these changes to prostaglandin production. The changes included: 1) increased breakdown of phosphatidylinositol (PI), 2) increased breakdown of diphosphoinositide (PI-P) and triphosphoinositide (PI-P2), 3) an increase in the mass of diglyceride and phosphatidic acid, 4) increased synthesis, first of PI-P and PI-P2, then phosphatidic acid, and, finally, PI, and 5) increased water-soluble inositol phosphates. Vasopressin treatment resulted in a significant increase (90%) in the mass of phosphatidic acid and a smaller (13%) decline in the mass of PI. Changes in diglyceride were seen following 45 s of treatment with vasopressin. Alterations of phosphoinositide metabolism were seen as early as 45 s and continued for up to 5 h following hormone exposure. By contrast, prostaglandin production declined after 30 min. These observations on vasopressin-stimulated metabolism in cultured mesangial cells may provide a basis for an understanding of the functional changes that follow hormone exposure. PMID- 2990240 TI - Phosphate uptake by proximal cells isolated from rabbit kidney: role of dexamethasone. AB - A single cell suspension was prepared from rabbit kidney cortex by gentle mechanical dissociation. The isolated cells were functionally intact and metabolically active, as indicated by exclusion of eosin dye, respiratory measurement, and ATP content. The isolated cells were shown to possess long microvilli, and their proximal origin was confirmed by enzymatic and glucose production studies. The uptakes of 0.1 mM phosphate (Pi) and 0.05 mM D-glucose at 37 degrees C were strongly dependent on the extracellular Na+. Incubation of the cells with parathyroid hormone (10 U/ml) for 20 min reduced 20-s Pi uptake by 20%. The addition of dibutyryl cAMP decreased Pi uptake, with a maximal effect at 10(-6) M. Incubation of cells with gluconeogenic substrates, under conditions in which glucose production was increased, did not promote any change in Pi accumulation. When incubated for 60 min at 37 degrees C in the presence of dexamethasone (Dex), the Na+-dependent Pi uptake was decreased by 29% at 10(-9) M and by 36% at 10(-7) M without modification of the glucose uptake. Replacing Dex by aldosterone (10(-7) M) remained without effect on Pi uptake. It is concluded that: renal cells isolated by our preparative method are mainly of proximal origin; isolated cells are a good model for studying the regulation of Pi uptake in the proximal tubule; and glucocorticoids have an acute specific effect on Pi transport detectable in vitro and this effect does not seem to be related to modifications of renal gluconeogenesis. PMID- 2990239 TI - Alpha2-adrenoceptors and cellular cAMP levels in single nephron segments from the rat. AB - A functional role for the numerically predominant renal alpha2-adrenoceptors, which in other tissues inhibit adenylate cyclase, remains undefined. We therefore examined the effect of alpha2-adrenoceptor stimulation with (-)-epinephrine (E) on cell cAMP content in the isolated proximal convoluted tubule (PCT), medullary and cortical thick ascending limb of Henle, and collecting tubule (MTAL, CTAL, MCT, and CCT, respectively). Parathyroid hormone (1-34 PTH), in PCT or CTAL, or arginine vasopressin (AVP), in MTAL, CTAL, MCT, or CCT, was used to activate adenylate cyclase in intact cells from these microdissected nephron segments in the presence of 3-isobutyl-1-methylxanthine (phosphodiesterase inhibitor) and propranolol. Alpha2-Adrenoceptors were activated using varying concentrations of E (37 degrees C, 2 min). Alpha2-Adrenoceptor activation with E (5 X 10(-7) to 5 X 10(-6) M) suppressed cellular cAMP stimulation by PTH by 35% in PCT and stimulation by AVP in CCT by 50%. This suppression by E in PCT and CCT was inhibited by 5 X 10(-6) M yohimbine or 5 X 10(-7) M phentolamine but not by 5 X 10(-6) M prazosin. E also suppressed cAMP stimulated by AVP in MCT, but it did not suppress the PTH-or AVP-stimulated increase in cellular cAMP in CTAL and MTAL. These studies show that there are alpha2-adrenoceptors in the rat nephron. Activation of these alpha 2-adrenoceptors can inhibit cAMP formation stimulated by PTH in PCT and by AVP in the CCT and MCT but not in the CTAL and MTAL. A pathophysiological role of altered regulation of these receptors is yet to be described. PMID- 2990241 TI - Evidence against role of antidiuretic hormone in support of blood pressure during dehydration. AB - The contribution of arginine vasopressin (AVP) to the maintenance of mean arterial pressure (MAP) during fluid deprivation was reinvestigated in chronically prepared, conscious, freely moving rats. In normal Long-Evans (LE) rats during water deprivation for up to 48 h plasma AVP levels increased from 2.98 +/- 0.62 to 17.55 +/- 2.03 pg/ml, but MAP and heart rate (HR) remained unchanged. Fluid deprivation for 10 h in Brattleboro homozygous rats that are unable to synthetize vasopressin resulted in a more severe water depletion than 48 h of dehydration in the normal rats, but also in this group MAP and HR were maintained at values not significantly different from control. Cardiac output (CO) fell during the course of dehydration in both LE and Brattleboro rats. Injection of 20 micrograms/kg of the vasopressin pressor antagonist, 1-deamino penicillamine, 2-(O-methyl)-tyrosine AVP [dPTyr(Me)AVP] did not decrease MAP or increase HR in LE rats after 24 h of water deprivation and was ineffective even after 48 h of fluid deprivation. CO also remained unchanged following AVP blockade. Efficacy of the antagonist was verified by complete inhibition of the pressor response elicited by exogenous AVP. The concentration of plasma catecholamines was unaltered by dPTyr(Me)AVP. Blockade of the renin-angiotensin system via converting-enzyme inhibition failed to reveal any depressor effect after injection of dPTyr(Me)AVP. Thus it appears that AVP is not involved in the maintenance of blood pressure after fluid deprivation in conscious rats. PMID- 2990242 TI - Time-dependent effect of insulin in suprachiasmatic nucleus on blood glucose. AB - Studies were made on the effects of insulin (30 or 300 microU) injected into the suprachiasmatic nucleus (SCN) of the hypothalamus on the plasma glucose and insulin concentrations in pentobarbital-anesthetized rats, which had been kept under 12:12 light-dark conditions. Insulin injection caused a decrease in the plasma glucose concentration in the light period but an increase in the dark period, these changes being significant within 2 min after injection. Insulin injection also caused a decrease in plasma insulin concentration in the light period but a tendency for its increase in the dark period. Intraperitoneal injection of insulin at the same dose did not affect the plasma glucose or insulin concentration in either the light or dark period. Insulin injection into the SCN decreased glycogen phosphorylase a activity but increased glycogen synthase I activity in the liver in the light period, but the insulin injection did not affect the activities of these enzymes in the dark period. Lesions including bilateral SCN completely eliminated the responses of the plasma glucose and insulin concentrations to insulin injection both in the light and dark periods. These findings suggest that insulin injection into the SCN causes time dependent changes in plasma glucose through changes in activities of glycogen metabolizing enzymes in the liver in the light period but through other mechanisms in the dark period. PMID- 2990243 TI - The ultimate stigma: inadequate funding for research on mental illness and addictive disorders. PMID- 2990244 TI - Hemorrhagic pancreatitis: a complication of transcatheter embolization treated successfully by total pancreatectomy. AB - Since pancreatitis can be produced experimentally in dogs by embolization of microspheres into the pancreatic arterial circulation, there has been speculation that intentional or inadvertent embolization of the pancreas in human subjects could also produce pancreatitis. Although such therapeutic embolization has increased, no pathologically documented case of this complication has been recorded. We have reported the first such case occurring in a patient with a large, highly vascular, nonfunctioning islet cell carcinoma of the tail of the pancreas preoperatively embolized with Gianturco coils and Gelfoam particles suspended in sodium tetradecylsulfate solution to facilitate distal pancreatectomy. The resultant hemorrhagic pancreatitis and duodenal necrosis required a total pancreatectomy. We conclude that, by itself, occlusion of the origin of the splenic and gastroduodenal arteries with coils would have been effective and without complication; however, the addition of Gelfoam particles in a sclerosing solution reduced the microscopic pancreatic circulation to a critical point and resulted in hemorrhagic pancreatitis. PMID- 2990245 TI - Perioperative ischemic injury after coronary bypass graft surgery. AB - Two hundred twelve patients who underwent isolated coronary bypass graft surgery were prospectively evaluated for perioperative ischemic injury. All patients underwent preoperative and postoperative testing with technetium 99m pyrophosphate first-pass ventriculography combined with myocardial uptake scans, 12-lead electrocardiography, and serial creatinine phosphokinase MB determination. Fifteen percent of the patients had ischemic injury with at least two test results positive, but only 4 percent had positive results of all three tests. No single test proved adequate. Enzyme levels were highly sensitive and had value as a screening test. The electrocardiogram was specific but only moderately sensitive. The single best test was the radionuclide scan with good sensitivity and no false-positive results. All three tests are required to rigorously diagnose ischemic injury. PMID- 2990246 TI - Breast carcinoma in situ. A retrospective review of 112 cases with a minimum 10 year follow-up. AB - We have retrospectively reviewed 112 cases of in situ carcinoma of the breast treated between 1960 to 1972, with a minimum 10 year follow-up to correlate treatment with outcome. We concluded that bilateral simple mastectomy with low axillary dissection is the treatment of choice for intraductal or lobular carcinoma in situ. This conclusion was based on the early age at diagnosis, the high incidence of bilaterality, and the long latency and probable progression from the in situ stage to the invasive stage. Lesser procedures can be endorsed for those patients of advanced age or who have associated medical problems whose life expectancy is estimated to be 10 years or less. Patients who refuse bilateral mastectomy should undergo biopsy of the involved or opposite breast at 3 to 5 year intervals regardless of physical findings or mammographic suspicions, especially when severe epithelial dysplasia is encountered in the biopsy specimens. This nonpalpable but potentially curable lesion remains difficult to detect even by mammography. PMID- 2990247 TI - Dissociated single cells from the inner ear: strial cells. AB - Using the enzyme dissociation technique, the authors successfully isolated strial marginal cells and intermediate cells from guinea pig and chinchilla cochleas. Dissociated cells were maintained for in vitro observation under the light microscope for up to 2 hours without gross evidence of cell damage. Such single cells were successfully prepared for scanning or transmission electron microscopic observation, which showed excellent cell shape and preservation of cell organelles. However, there was evidence of increased vesiculation in the dissociated marginal cells, which is interpreted to be evidence of increased pinocytotic activity. PMID- 2990248 TI - [Mechanism of action of anti-D-gamma globulin in preventing rhesus alloimmunization in the pregnant woman and hemolytic disease of the newborn infant]. PMID- 2990249 TI - [Anesthesiologic aspects of chemonucleolysis in local anesthesia]. AB - After treatment of herniation of a lumbar disc by injection of the enzyme chymopapain, i. e. after chemonucleolysis, anaphylactic reactions can occur in about one per cent of the cases. In order to recognise the pattern of signs associated with such reactions, well in advance, while avoiding the additional risk of general anaesthesia, some authors propagate local anaesthesia. We report on our perioperative procedure in 102 cases of chemonucleolysis under local anaesthesia. Prick's tests were carried out before surgery to exclude sensitization to the substances to be injected. In two cases only due to a positive prick test to chymopapain chemonucleolysis had to be effected with collagenase; as a matter of fact, collagenase is not known to have caused any anaphylactic reactions, but it may be responsible for local side effects, such as destruction of adjacent tissues. The patients were kept under observation by an anaesthetist during and after surgery. No anaphylactic reaction was seen. Chemonucleolysis appears to be a suitable treatment method provided it is carried out under local anaesthesia with the same precautions as applied under regional anaesthesia by the anaesthetist. PMID- 2990250 TI - [Acid-base homeostasis in anesthesiology and operative intensive care medicine. Physiology, pathophysiology and clinical aspects of acidoses]. AB - Basic and clinical aspects of acid-base physiology and acidoses are reviewed in this paper. Therapeutic principles on the treatment of metabolic and respiratory acidosis are discussed in detail. As far as buffer substances are necessary for antiacidotic therapy, sodium bicarbonate seems to be the agent of choice today. Sodium bicarbonate has a better extra- and intracellular effect than THAM, and should therefore be preferred in every day practice. PMID- 2990251 TI - Purification of nonspecific protease-free collagenase from Clostridium histolyticum. AB - The collagenase (EC 3.4.24.3) produced by the bacterium Clostridium histolyticum has been purified free from nonspecific protease contaminants by a two-step procedure. The crude culture medium is chromatographed over heparin-Sepharose and Sephacryl S-200, and the resulting preparation has no activity versus noncollagenous proteins or N alpha-benzoyl-L-arginine ethyl ester, yet cleaves native thermally reconstituted collagen fibrils quite efficiently (specific activity, 3000 units/mg). The purification described may be useful for those investigators requiring substantially purified collagenase for applications such as cell culture or collagen quantitation in protein mixtures. PMID- 2990252 TI - An assay for ribonucleotide reductase based on ion-exchange chromatography of the reaction product. AB - A rapid and convenient assay for ribonucleotide reductase has been developed in which the reaction product, deoxycytidine diphosphate (dCDP), is isolated without further conversion. The enzymatic reaction is terminated by the addition of ethanol and the sample is chromatographed on a single, small, and disposable column of polyethylenimine cellulose. A two-step elution is conducted with buffers containing 25% ethanol. First, contaminants and byproducts such as cytidine and its monophosphate are removed at low ionic strength while the diphosphates are retained. Then dCDP is selectively eluted as a sharp peak with a strong borate buffer. Under these conditions, the excess substrate, cytidine diphosphate, remains on the column, presumably as the borate complex. The assay is linear with time for 15 min at 25 degrees C and linear with the amount of enzyme even at very low concentrations. With slight modifications, the assay seems applicable to the use of UDP or ADP as substrates. The method is not suitable for samples which contain nucleotide kinase or other interfering enzymes which convert a significant amount of dCDP into byproducts. However, another chromatographic system based on similar principles has been found which could be used to measure any dCTP produced in this way. PMID- 2990253 TI - Separation of the major adrenal steroids by reversed-phase high-performance liquid chromatography. AB - The major adrenal steroids were separated by multistep gradient elution with a reversed-phase high-performance liquid chromatography system, employing water and 1-propanol as solvents. With this solvent system, a wide range of 21 5-ene-3 beta ol and 4-ene-3-one steroids can be resolved in a single chromatogram, which was not possible with previously published gradient solvent systems. In particular, intermediate steroids of the biosynthetic pathway, 17 alpha-hydroxyprogesterone, 17 alpha-hydroxypregnenolone, and dehydroepiandrosterone, were separated with baseline or sufficient resolution to allow accurate quantitation. Using the 1 propanol-water gradient, the separations of 5-ene and 4-ene steroids were compared on different octadecylsilyl packings. Optimum resolution was obtained with a fully covered, spherical particle. The 1-propanol-water gradient was compared to a previously published methanol-water gradient in the analysis of steroidogenesis by adrenocortical cell cultures. HPLC analysis of the steroid production was quantitatively the same with both gradient solvent systems. However, qualitatively, the methanol-water gradient system did not resolve the above-mentioned intermediate steroids. PMID- 2990254 TI - Spectrophotometric assay for vertebrate collagenase. AB - Collagenase from normal human skin fibroblasts was found to catalyze the hydrolysis of esters and thio esters. This observation led to the development of a rapid, sensitive, continuous spectrophotometric assay for vertebrate collagenase using the thio peptolide Ac-ProLeuGly-S-LeuLeuGly-OC2H5 as substrate in the presence of 4,4'-dithiodipyridine or Ellman's Reagent. A Km of 0.004 M and a kcat of 370,000 h-1 were determined for the thio peptolide-enzyme reaction. The method is able to detect collagenase at concentrations as low as 2 ng/ml. PMID- 2990255 TI - Assay of covalent intermediate of 5'-nucleotide phosphodiesterase. AB - We report a new procedure for isolating a covalent phosphoryl enzyme (diester) intermediate of bovine intestinal 5'-nucleotide phosphodiesterase. The convenience of the procedure makes it possible to determine effects of reaction conditions on the yield of covalent intermediate. Under optimum conditions, using [methyl-3H]deoxythymidine 5'-triphosphate as substrate, more than 50% of the enzyme is recovered as thymidylyl enzyme, a 10-fold increase in yield over the previous procedure (M. Landt and L. G. Butler, 1978, Biochemistry 17, 4130-4135). Yields of thymidylyl enzyme were maximal at pH 4, whereas optimum catalytic activity is observed at pH greater than 9. PMID- 2990256 TI - Proteolytic release of glycopeptides from glycoproteins transferred to nitrocellulose following gel electrophoresis. AB - To determine whether glycopeptides could be released from glycoproteins bound to nitrocellulose, the glycoproteins of murine mammary tumor virus (MuMTV) were radiolabeled by the periodate oxidation/tritiated sodium borohydride reduction technique and separated by gel electrophoresis followed by diffusion transfer. Pronase digestion of nitrocellulose filter strips containing labeled glycoproteins (gp55 or gp34) revealed a rapid release of glycopeptides, i.e., approximately total release within 4 h. The released glycopeptides were similar in size, as determined by molecular sieving chromatography, to glycopeptides obtained by proteolytic digestion of MuMTV glycoproteins from dried gel strips (A. Zilberstein et al., 1980, Cell 21, 417-427) or in solution (M. J. Yagi et al., 1978, Virology 91, 291-304). PMID- 2990257 TI - The analyses of six common vitamins by laser desorption mass spectroscopy. AB - The ability to obtain molecular fragmentation patterns at localized (microprobe level) sites would be of great aid to analytical biochemists in mapping the sites, and thus deducing the many functions of bioorganic species, such as vitamins. Laser desorption mass spectroscopy has this ability to provide molecular fragmentation patterns at the microprobe level. In this preliminary study, six common vitamins--ascorbic acid, retinoic acid, pyridoxine monohydrochloride, riboflavin, thiamine hydrochloride, and cholecalciferol--are analyzed as pure compounds by laser desorption mass spectroscopy. Spectra and commentary about the fragmentation patterns for all six vitamins are included. PMID- 2990258 TI - Effects of solvent composition and temperature on the separation of anilines with silica, amino, and diamine bonded phase columns. PMID- 2990259 TI - The role of eicosanoids in respiratory mucus hypersecretion. AB - The role of eicosanoids in the excessive secretion of respiratory mucous glycoproteins (MGP) accompanying immediate hypersensitivity and inflammatory pulmonary states has only been addressed in the last few years. Three lines of evidence suggest that eicosanoids may participate in the physiologic regulation of MGP secretion as well as being capable of stimulating increased MGP production. First, inhibition of eicosanoid generation with corticosteroids or eicosatetraynoic acid (ETYA) reduces ongoing baseline MGP secretion while selective inhibition of prostaglandin production with nonsteroidal anti inflammatory agents increases MGP release. Second, arachidonic acid and a variety of eicosanoids stimulate MGP secretion in vitro. In fact, several lipoxygenase pathway metabolites including hydroxyeicosatetraenoic acids (HETEs) and leukotrienes (LTC4 and LTD4) significantly increase MGP secretion in nanogram quantities. Third, a variety of pulmonary cells including airway epithelium, endothelial cells, mast cells, alveolar macrophages and neutrophilic leukocytes generate eicosanoids under conditions that would be encountered in clinical states in which mucus secretion occurs. Thus, mucus secretion during both normal and stimulated states would be influenced by eicosanoids. It seems likely that this eicosanoid-mucus secretion relationship is very important in all forms of asthma, but most particularly in aspirin-related asthma. PMID- 2990261 TI - Evaluation of acute scrotum pain with testicular scanning. AB - Emergency room physicians frequently are faced with the evaluation of the patient with acute testicular pain. The rapid differentiation between testicular torsion and acute epididymitis is necessary although frequently difficult due to an atypical presentation. We describe four patients with testicular pain in whom testicular scanning provided useful adjunctive information regarding diagnosis. This technique clearly differentiates acute torsion, acute epididymitis, testicular rupture, and missed torsion with abscess formation, prompting appropriate therapy in each case. Testicular scanning provides a rapid, noninvasive method to assist in the evaluation of testicular pain. PMID- 2990260 TI - [Electro-drug anesthesia. Clinical and hormonal effects of transcranial electrostimulation]. AB - The French technique of anaesthesia by electrostimulation described in 1972 by Cara and coworkers, consists of transcranial electrostimulation by means of a high frequency current combined with administration of a neuroleptic drug, a benzodiazepine, a curare and nitrous oxide with oxygen. Fentanyl is also given by some authors. In order to assess the benefit of such electrostimulation, this study compared two randomized groups of ten patients, scheduled for abdominal and pelvic surgery. Both groups received the same drugs (i.e. droperidol, flunitrazepam, pancuronium and nitrous oxide with oxygen), whereas patients in group I were also submitted to electrostimulation. This study describes and discusses the clinical behaviour of patients and the hormonal reactions before, during and after surgery. In both groups, operative conditions were satisfactory. Recovery and onset of spontaneous ventilation were rapid and no patient had an unpleasant recall of the operation itself. However, most of them complained of postoperative pain. Electrostimulation did not reduce the quantity of drugs required during and after surgery. In both groups, circulatory activity was significantly increased. In group I, the arterial pressure and the heart rate were significantly higher than in group II during and after surgery. The hormonal reactions showed that in both groups adrenocorticotrophic hormone, growth hormone and antidiuretic hormone increased during surgery. Adrenocorticotrophic hormone concentration was higher in group I during the operation. The serum levels of cortisol decreased before surgery in group I and rose in both groups during and after laparotomy; prolactin increased before surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990262 TI - Factors associated with the spread of clinical vesicular stomatitis in California dairy cattle. AB - In 1983, vesicular stomatitis entered the state of California through infected cattle purchased in Idaho and sold to 5 California dairies. This study examined management, environmental, and host factors which were thought to be associated with disease spread. The use of coarse roughage and hard-pelleted concentrates, the presence of uneaten feed potentially contaminated with virus-laden saliva, increased interpen movement of cows, poor ground surface conditions, poor milking hygiene, and poor teat sanitation were management factors observed to be associated with clinical signs. The amount of milk production, age, or days in milk of individual cows seemed to have an important role in the spread of clinical disease. Recommendations are given for control and prevention of clinical signs and, therefore, the severity of disease during epizootics of vesicular stomatitis in California dairies. PMID- 2990263 TI - Blood from bovine leukemia virus-infected cattle: antigen production correlated with infectivity. AB - Leukocytes from 1 ml of blood from cattle seropositive to the bovine leukemia virus were cultured for 3 days and then tested by radioimmunoassay for antigen production. Infectivity of blood from each animal was also tested by calf inoculation and subsequent serologic detection of bovine leukemia virus transmission by agar-gel immunodiffusion. In a preliminary experiment, blood from each of 3 antigen-positive cattle was inoculated intracutaneously into 2 calves in volumes of 20 or 100 microliter. Blood from each of 4 antigen-negative cattle was similarly inoculated into 3 calves in volumes of 20, 100, or 500 microliter. At the termination of the experiment (8 weeks after inoculation), all 6 calves given blood from antigen-positive cattle had seroconverted, and 11 of 12 calves given blood from antigen-negative cattle had seroconverted. In a 2nd experiment, blood from each of 2 antigen-positive and 2 antigen-negative cattle was inoculated into pairs of calves in volumes of 1, 10, or 20 microliter. At the end of the experiment (12 weeks after inoculation), all calves inoculated with blood from antigen-positive cattle had seroconverted, but only 6 of 12 calves that had been given blood from antigen-negative cattle had seroconverted. The relative infectivity of blood was best illustrated by comparing results from the 1 microliter inoculations. At that volume, the 4 calves given blood from antigen positive cattle were infected, whereas none of 4 calves given blood from antigen negative cattle was infected. PMID- 2990264 TI - Rapid enzyme-linked immunosorbent assay for detecting antibodies to canine parvovirus. AB - A rapid screening assay for determining antibodies to canine parvovirus in dog serum using monoclonal antibodies and enzyme-linked immunosorbent assay (ELISA) technology was developed. The ELISA could be read visually, and the results correlated well with serum neutralization (SN) and hemagglutination inhibition (HI) titers. Sera with SN less than or equal to 1:4 or HI less than or equal to 1:10 had an 87.9% correlation with ELISA and sera with SN greater than or equal to 1:64 or HI greater than or equal to 1:80 had a 94.4% correlation. The assay took only 10 to 15 minutes to perform and did not require specialized equipment. The ELISA should be useful in monitoring dogs for the presence of maternal antibodies against parvovirus and for determining seroconversion after vaccination. PMID- 2990265 TI - Release of immunosuppressive substances from tissue culture cells infected with bovine viral diarrhea virus. AB - Bovine fetal lung tissue culture cells were infected with bovine viral diarrhea virus. The cells released substances into the supernatant fluid that suppressed the proliferative response of bovine peripheral blood mononuclear cells which had been stimulated with concanavalin A. Activity could be found in low molecular weight fractions (less than or equal to 2,000 daltons). During the assay, these fractions were required to inhibit the response of leukocytes to concanavalin A. Addition of indomethacin to the infected tissue culture cells inhibited generation of immunosuppressive supernatants; addition of indomethacin to the suppressive supernatants did not block their activity. The possible involvement of prostaglandins in this immunosuppressive activity is discussed. PMID- 2990266 TI - Bovine herpesvirus-1 infection of cattle: kinetics of antibody formation after intranasal exposure and abortion induced by the virus. AB - The kinetics of antibody formation in Holstein heifers after primary and secondary intranasal inoculation of bovine herpesvirus-1 (BHV-1) and after BHV-1 induced abortion was determined. Sera were fractionated by gel filtration and ion exchange chromatography. The antibody activity within serum immunoglobulin (Ig) isotypes was assessed, using a plaque-reduction neutralization assay. The primary immune response to BHV-1 infection was characterized by the appearance of IgM and IgG antibodies in serum by postinoculation day (PID) 7. Maximal IgG antibody activity occurred at PID 35 in nonpregnant heifers and at PID 14 in pregnant heifers. Thereafter, IgG antibody activity declined slowly in both groups of heifers. Maximal IgM antibody activity occurred at PID 14 in both groups of heifers and declined rapidly thereafter. The IgG antibody activity during primary immune responses was restricted to the IgG1 subclass. Secondary responses were characterized by anamnestic IgG antibody responses. Antibody activity was present within the IgG1 and IgG2 subclasses during secondary immune responses, but the increase in antibody activity during this period was primarily in the IgG2 subclass. Secondary IgM antibody formation was elicited by abortion induced by the intra-amniotic inoculation of BHV-1, but not by reexposure by the intranasal route. Abortion occurred in 1 heifer 28 days after intranasal BHV-1 inoculation. Abortion in this heifer was not associated with a secondary antibody response. The nature of BHV-1 antigenic exposure in the bovine determined the relative distribution of anti-BHV-1 antibody activity in serum IgM, IgG1, and IgG2. The formation of IgM antibody, with the exception of secondary intranasal exposure, indicated recent BHV-1 antigenic exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990267 TI - Kinetics of antibody formation after the reactivation of bovine herpesvirus-1 infection in cattle. AB - Holstein heifers previously infected with bovine herpesvirus-1 were treated with corticosteroids to reactivate latent infection. The kinetics of serum immunoglobulin (Ig) M, IgG1, and IgG2 antibody formation were determined. The secondary immune response which developed as a consequence of virus reactivation was characterized by an anamnestic IgG antibody response. Secondary IgG1 and IgG2 antibody formation constituted the IgG antibody response. In addition, secondary IgM antibody formation was elicited by this form of bovine herpesvirus-1 antigenic exposure. PMID- 2990269 TI - Adenosine-induced bronchoconstriction in asthma. Role of parasympathetic stimulation and adrenergic inhibition. AB - Adenosine by inhalation causes bronchoconstriction in asthmatic but not in normal subjects by an undefined mechanism. This study investigated the roles of cholinergic reflex stimulation and decreased beta 2-adrenoceptor responsiveness to explain adenosine's bronchoconstrictor action. The protection afforded by the inhaled muscarinic cholinergic antagonist, ipratropium bromide (IB) 1 mg, from bronchoconstriction induced by inhaled adenosine was compared with that of methacholine in 8 allergic asthmatic subjects. After saline placebo, the geometric mean concentrations of adenosine required to produce a 20% fall in FEV1 (PCf20) was 2.20 mg/ml and a 35% fall in SGaw (PCs35) was 1.97 mg/ml, which compared to 0.13 and 0.11 mg/ml, respectively, for methacholine. The IB increased FEV1 by 11 to 15% and SGaw by 69 to 73% and provided a large degree of protection against methacholine, with a geometric mean concentration ratio (CR) of 196 when airway caliber was measured as SGaw (p less than 0.001). In contrast, IB provided little protection against adenosine-induced bronchoconstriction (CR 1.3 for SGaw and 1.51 for FEV1). Beta 2-adrenoceptor responsiveness of the airways after inhaled adenosine and histamine was further studied in 12 asthmatic subjects by observing the antibronchoconstrictor effect of inhaled isoproterenol. After equivalent degrees of bronchoconstriction, 35 to 36% fall in FEV1 and 60 to 62% fall in SGaw cumulative doses of inhaled isoproterenol produced almost identical maximal increases in FEV1 and SGaw after adenosine as achieved after histamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990268 TI - Nucleotide sequences of the genes for Escherichia coli heat-stable enterotoxin I of bovine, avian, and porcine origins. AB - The nucleotide sequences of cloned Escherichia coli heat-stable toxin 1 (STI) genes isolated from bovine, avian, and porcine origins were determined. They were found to be almost identical to that of Tn1681. The nucleotide sequences were completely preserved in bovine and avian genes, whereas the porcine gene had different sequences at 3 positions in the external region of STI structural genes, as compared with Tn1681. The amino acid sequences of the STI genes of the 3 animal origins corresponded to STIa, which had initially been found in a bovine strain. PMID- 2990270 TI - Growth factors, oncogenes, and lung cancer. PMID- 2990271 TI - Prediction of therapeutic response in steroid-treated pulmonary sarcoidosis. Evaluation of clinical parameters, bronchoalveolar lavage, gallium-67 lung scanning, and serum angiotensin-converting enzyme levels. AB - To find a pretreatment predictor of steroid responsiveness in pulmonary sarcoidosis we studied 21 patients before and after steroid treatment by clinical evaluation, pulmonary function tests, bronchoalveolar lavage (BAL), gallium-67 lung scan, and serum angiotensin-converting enzyme (SACE) level. Although clinical score, forced vital capacity (FVC), BAL percent lymphocytes (% lymphs), quantitated gallium-67 lung uptake, and SACE levels all improved with therapy, only the pretreatment BAL % lymphs correlated with the improvement in FVC (r = 0.47, p less than 0.05). Pretreatment BAL % lymphs of greater than or equal to 35% predicted improvement in FVC of 10/11 patients, whereas among 10 patients with BAL % lymphs less than 35%, 5 patients improved and 5 deteriorated. Clinical score, pulmonary function parameters, quantitated gallium-67 lung uptake, and SACE level used alone, in combination with BAL % lymphs or in combination with each other, did not improve this predictive value. We conclude that steroid therapy improves a number of clinical and laboratory parameters in sarcoidosis, but only the pretreatment BAL % lymphs are useful in predicting therapeutic responsiveness. PMID- 2990272 TI - Alteration of bronchoalveolar cells during murine cytomegalovirus interstitial pneumonitis. AB - In BALB/c mice, murine cytomegalovirus (MCMV) in conjunction with a single dose of cyclophosphamide (CP) induces a diffuse interstitial pneumonitis not seen with either virus or CP alone. To gain insight into the host immune mechanisms operating in the lung during interstitial pneumonitis, we examined the cells recovered in bronchoalveolar lavage (BAL) fluids of mice with MCMV with and without CP. During MCMV interstitial pneumonitis, there was a significant increase in the total BAL cells recovered, primarily because of an influx of lymphocytes bearing the Thy 1.2 marker. Although the number of cells with Lyt 1 and Lyt 2 markers increased, the most significant increase was in the proportion of lymphocytes with surface asialo-GM1. Delineating the roles of these various cell populations may provide insight into the pathogenetic mechanisms leading to CMV interstitial pneumonitis. PMID- 2990273 TI - [Chronic idiopathic hypoparathyroidism: report of 3 cases]. AB - We report three patients presenting symptoms of tetany at eleven months, five years and seven years. They had hypocalcemia hypomagnesemia, hyperphosphatemia and elevated phosphate tubular reabsorption. PTH serum levels were decreased and cAMP urinary excretion was very low. Clinical and biological findings suggest hypoparathyroidism, but due to peculiar phenotype of one patient differential diagnosis is established. PMID- 2990274 TI - [Papular acrodermatitis of childhood as Gianotti-Crosti syndrome]. AB - A case of a boy with a papular acrodermatitis caused by an Epstein-Barr viral infection is presented. This makes us consider actualized criteria relating to diseases in the Gianotti-Crosti syndrome and its relationship with the presence or absence of Australian antigen. PMID- 2990275 TI - The acquired immunodeficiency syndrome in a cohort of homosexual men. A six-year follow-up study. AB - A cohort of 6875 homosexual men, initially seen at the San Francisco City Clinic between 1978 and 1980, were studied to determine the incidence and prevalence of the acquired immunodeficiency syndrome, related conditions, and infection with the human T-lymphotropic virus, type III/lymphadenopathy-associated virus (HTLV III/LAV). By December 1984, 2.4% of the men had the syndrome; mortality attributable to the syndrome in 1984 was 600/100 000. For each man with the syndrome in a representative sample of 474 cohort members seen in 1984, 7.5 men had generalized lymphadenopathy, 1.1 had other prodromal findings, and 0.8 had hematologic abnormalities. Prevalence of serum antibodies to HTLV-III/LAV, measured by an enzyme-linked immunosorbent assay, increased from 4.5% in 1978 to 67.4% in 1984. Of 31 persons who were seropositive and without the syndrome between 1978 and 1980, 2 developed the syndrome and 8 developed related conditions during a median follow-up of 61 months. Over a 6-year period, two thirds of cohort members were infected with HTLV-III/LAV and almost one third developed related conditions. PMID- 2990277 TI - C-type retrovirus-related P-30 protein in human follicular fluids: relation to follicular steroids. PMID- 2990278 TI - Clomiphene-induced changes in endometrial receptor kinetics on the day of ovum collection after ovarian stimulation: a study on cytosol and nuclear estrogen and progestin receptors and 17 beta-hydroxysteroid dehydrogenase. PMID- 2990276 TI - Hemolytic anemias and erythrocyte enzymopathies. AB - The human erythrocyte generates high-energy adenosine triphosphate by anaerobic glycolysis and cycles oxidized and reduced nicotinamide adenine dinucleotide phosphate by the aerobic pentose phosphate shunt pathway. Certain enzymopathies of the pentose phosphate shunt are associated with hemolysis resulting from oxidative denaturation of hemoglobin. Glucose-6-phosphate dehydrogenase deficiency, an X-chromosome-linked disorder, is the prototype of these diseases and is genetically and clinically polymorphic. Six enzymopathies of anaerobic glycolysis cause hemolytic anemia; lactate dehydrogenase deficiency does not. In 2,3-diphosphoglycerate mutase deficiency, 2,3-diphosphoglycerate is greatly reduced and asymptomatic polycythemia is noted. Pyrimidine-5'-nucleotidase deficiency, an enzymopathy of nucleotide metabolism, is characterized by intracellular accumulations of pyrimidine-containing nucleotides, marked basophilic stippling on the stained blood film, splenomegaly, and hemolysis. Lead inhibits the nucleotidase and an identical syndrome occurs during severe lead poisoning. Hemolysis also accompanies an unusual enzymopathy characterized by a 40- to 70-fold increase (not decrease) in adenosine deaminase activity. PMID- 2990279 TI - Oocyte maturation as regulated by follicular factors. AB - Follicular fluid (FFl), obtained from 24 women treated with clomiphene/hCG in an in vitro fertilization program, was characterized with respect to steroid hormone levels and oocyte maturation inhibitor (OMI) activity. Three FFl samples apparently were derived from cystic follicles and contained low steroid levels and no OMI activity in an in vitro rat oocyte assay. The remaining 21 follicles contained normal preovulatory steroid levels and mature and generally fertilizable oocytes. In 7 of these follicles the FFl (at 50% concentration) significantly inhibited rat oocyte meiosis, while 14 exerted no OMI activity. The results confirm earlier work on porcine and human FFl, suggesting that the putative OMI activity declines with follicular maturation. PMID- 2990280 TI - Sexual dimorphisms as an index of hormonal influences on conditioned food aversions. PMID- 2990282 TI - Direct and indirect behavioral pathways to the social transmission of food avoidance. PMID- 2990281 TI - Glucocorticoid and other hormonal substrates of conditioned taste aversion. AB - Results of research reported in this paper show that lithium chloride (LiCl) injection causes a marked and prolonged elevation of adrenocorticotropic hormone (ACTH) and corticosterone. Peak elevations occur within 30 min after injection and continue for 60 to 120 min depending upon the molarity of the LiCl solution. CS preexposures do not alter the pituitary-adrenal response to LiCl. Although this response occurs, evidence argues that pituitary-adrenal activity is neither a sufficient nor essential condition for CTA to occur. It is possible that this prolonged ACTH secretion, which would be essential to maintain corticosterone levels elevated for one to two hours, is acting as a component of the unconditioned response associated with LiCl injections. Under certain types of extinction (forced extinction) the P-A system is activated. Thus, when an animal drinks an aversive solution following deprivation, intake of the solution causes a conditioned elevation of plasma corticosterone. Two measures of CTA, behavioral and hormonal, do not always coincide with one another. Preexposure to the CS alters both behavioral and corticosterone indices of CTA. These two systems will show either a coupling (e.g., parallel change) or a dissociation depending on the number of preexposures prior to conditioning. Manipulating the hormones associated with the pituitary-adrenal system also affects the acquisition of CTA and influences extinction. Thus, if one pretests with dexamethasone phosphate (DEX) prior to the administration of LiCl, CTA is attenuated. It would appear that ACTH is involved in the acquisition of CTA since a central block of ACTH also affects the magnitude of the aversion. If ACTH is injected during recovery, animals show a prolonged suppression of drinking. These data appear to be best explained using a memory-retrieval model. The manipulation of hormones and their effects on CTA appear to follow the effects observed endogenously. Thus, ACTH administered to rats during the conditioning phase does not appear to have any effect upon the learning of the aversion. Since LiCl markedly elevates ACTH, as indicated by prolonged elevations of corticosterone, the additional ACTH given exogenously at the time of conditioning does not appear to add to the effects already attributable to endogenous ACTH. Conversely, DEX given during recovery appears to have no effect upon the recovery function. Because endogenous levels of ACTH are already low during the free extinction procedure, DEX treatment does not appear to further reduce ACTH and has no effect upon recovery.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990284 TI - Conditioned taste aversions: a behavioral index of toxicity. PMID- 2990283 TI - Drug interactions measured through taste aversion procedures with an emphasis on medical implications. PMID- 2990285 TI - Conditioned food aversions and satiety signals. PMID- 2990286 TI - Learned food aversions in the progression of cancer and its treatment. AB - The studies included in this chapter examine the learned food aversions that develop as a result of cancer and cancer treatment. Clinical studies have shown that cancer patients can develop learned aversions to a novel ice cream flavor when it is consumed before drug treatments that produce nausea and vomiting. They also provided evidence that patients can acquire aversions to food in their usual diets when these foods are eaten before similar drug treatments. Observations in the clinic, supported by complementary studies with animal models, suggest that learned aversions are more likely to arise to protein foods than to other nutrient sources and that the presentation of a novel food in association with drug treatments may act as a "scapegoat" in blocking the development of aversions to foods in the normal diet. Laboratory studies using transplantable tumors in rats have shown that tumor growth can be associated with the development of strong aversions to the available diet. These aversions are specific to the diet eaten during tumor growth and they appear to play a causal role in the development of tumor-induced anorexia. The food aversions apparent in animals with certain experimental tumors point to physiological consequences of tumor growth that act as unconditioned stimuli in taste aversion conditioning. The identification of these changes and development of methods for correcting them are the current goals of our research in this area. PMID- 2990287 TI - Introduction: physiological mechanisms in conditioned taste aversions. PMID- 2990289 TI - Role of beta-endorphin in behavioral regulation. PMID- 2990288 TI - Peripheral and central adrenergic influences on brain systems involved in the modulation of memory storage. PMID- 2990290 TI - Noradrenergic modulation of selective attention: its role in memory retrieval. PMID- 2990291 TI - Phosphoprotein regulation of memory formation: enhancement and control of synaptic plasticity by protein kinase C and protein F1. PMID- 2990292 TI - Catecholamine system involvement in age-related memory dysfunction. PMID- 2990293 TI - The cholinergic hypothesis: a historical overview, current perspective, and future directions. PMID- 2990294 TI - Models of memory dysfunctions. PMID- 2990295 TI - Locus coeruleus as a target for psychogeriatric agents. PMID- 2990296 TI - Mediation of storage and retrieval with two drugs that selectively modulate serotonergic neurotransmission. PMID- 2990297 TI - Pharmacological manipulation of the substantia innominata-cortical cholinergic pathway. PMID- 2990298 TI - Abnormal globin gene structure and expression in beta-thalassemia. AB - Over the past five years, several new defects in the beta-thalassemias have been described from this laboratory using both restriction enzyme and sequencing analyses of cloned beta-thalassemia genes. The enzyme HphI has been shown to recognize a single nucleotide change at the 5' end of beta-IVS 2, and, using restriction enzyme analysis, demonstrated for the first time a specific defect associated with beta(0)-thalassemia. Cloning and sequencing of a beta-thalassemia gene have identified a single base change within IVS 2 at a position 705 nucleotides from the 5' end of IVS 2 that results in a beta(0)-thalassemia phenotype; no normal splicing occurs in this gene despite the fact that both the 5' and 3' ends of IVS 2 are unchanged. A unique and strong cryptic 3' acceptor splice site present in the normal gene at a position 580 nucleotides from the 5' end is used extensively in the mutant gene. Studies of this gene have indicated that there are sequences within IVS that are responsible for optimal expression of this gene; changes in these sequences can lead to markedly abnormal patterns of splicing. In addition, beta-globin gene expression has been evaluated in human erythroleukemia cells, K562 cells, and, although stable transformants with integrated beta-globin genes have been obtained, none of these transformants expressed the added beta-globin genes. This is presumably due to trans-acting factors or distal cis-acting effects that suppress the expression of these added beta-globin genes. In addition, a low epsilon-producing cell line, Bos cells, was used as a recipient for an exogenous epsilon-globin gene. A neomycin resistance gene was cotransfected into these cells, and a neomycin analogue (G418) was used to select cells containing both the neomycin resistance and epsilon-globin genes. Using Southern blotting, 10 of 11 stably transformed G418-resistant lines, which contain intact epsilon-globin genes, express epsilon-globin mRNA at much higher levels than the Bos cells into which they were transfected. Two of these lines express the epsilon-globin genes at a level comparable to that of wild-type K562 cells. These results indicate that the transfer and expression of human globin genes in human erythroid cells is feasible, and can occur at a high level.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990299 TI - Deletion analysis of the human gamma-globin IVS2: sequence requirements for RNA splicing. PMID- 2990300 TI - A novel deletion in the beta-globin gene complex. PMID- 2990302 TI - Prenatal diagnosis of hemoglobinopathies by DNA analysis. PMID- 2990301 TI - Analysis of globin gene inversion reveals similarities with immunoglobulin rearrangement. PMID- 2990303 TI - First trimester diagnosis of the hemoglobin disorders. PMID- 2990304 TI - Molecular rearrangements of the human alpha-globin gene cluster. PMID- 2990305 TI - Metabolism of structurally abnormal mRNAs resulting from beta-thalassemia mutations. PMID- 2990306 TI - Liposomes as vectors for calcium ionophores. PMID- 2990308 TI - The biology of Down syndrome. PMID- 2990307 TI - Liposome-embedded iron-porphyrins as an artificial oxygen-carrier. PMID- 2990309 TI - Use of restriction fragment length polymorphisms to study the origin of human aneuploidy. PMID- 2990310 TI - Genetic linkage map for chromosome 21. PMID- 2990311 TI - Linkage map on chromosome 21q and the association of a DNA haplotype with a propensity to nondisjunction and trisomy 21. PMID- 2990312 TI - [ORL localizations of Abrikosov's tumors. Apropos of 5 cases. A tumor which is not always benign]. AB - Abrikossoff's tumor is a benign lesion affecting the oropharynx in more than half the cases. Five cases are reported and symptoms and signs reviewed. Treatment was by wide excision, although a less extensive operation is possible to avoid major damage to surrounding structures. This attitude can be adapted because the course of the affection is by slow progression, with the possibility of no further growth, and the fact that surgery can be repeated. PMID- 2990314 TI - [Malignant eccrine poromas. Ultrastructural study of 2 cases]. PMID- 2990313 TI - Adenoid cystic carcinoma of the nasopharynx. Case reports and literature review. AB - Adenoid cystic carcinoma of the nasopharynx is a rare disease with 34 previously reported cases in the literature. We report three additional cases with clinical presentation, treatment, and outcome. The presenting symptoms were facial pain in one case, middle ear effusion and epistaxis in another, and diplopia in the third. Radiotherapy was administered with relief of symptoms in all three cases. However, two patients with long-term follow-up developed local recurrence as well as distant metastases. The literature review showed that patients with this disease present with symptoms and signs such as insidious onset of facial pain, epistaxis, diplopia, and hearing loss. The final outcome, despite vigorous treatment, was dismal. PMID- 2990315 TI - [Extra-mammary Paget's disease]. PMID- 2990316 TI - Radionuclide cystography in children: comparison of direct (retrograde) and indirect (intravenous) techniques. PMID- 2990317 TI - Effect of dexamethasone on the mean plaque size of bovine herpesvirus 1. AB - The effect of dexamethasone on infectious bovine rhinotracheitis virus (Bovine herpesvirus 1: BHV 1) was studied by measuring the mean plaque size produced by eight virus strains under two concentrations of dexamethasone phosphate (0.1 and 1 mM). Dexamethasone induced a significant reduction of the mean plaque size, whatever the strain used. Some differences were noted between the BHV 1 strains studied, depending on the dexamethasone concentration. PMID- 2990318 TI - Constitutional mosaic t(2;7)(q33;p22) and other rearrangements in a girl with Wilms' tumor. AB - A 2-year-old girl with sporadic unilateral Wilms' tumor (WT) not associated with aniridia was found to have, besides other chromosome abnormalities, a t(2;7)(q33;p22) in 6% of her lymphocytes. A comparison with 7 previous WT cases without aniridia in whom diverse chromosomal aberrations were present, reveals a wide heterogeneity and lead us to tentatively classify such changes as causal, secondary, and casual. PMID- 2990319 TI - [Reconstructive microsurgery of the vas deferens. Experimental study of suture materials]. AB - Microsurgery is the procedure of choice for vasectomy reversal. The aim of this study was two compare two different suture materials for vasovasectomy - a nonresorbable material (nylon 10/0 with a BV 6 needle), which is widely used, and a resorbable material (polyglycolic acid, also with a BV 6 needle), which has not yet been evaluated for this use. 28 Sprague-Dawley rats were operated on under microscope. Two groups were then compared, group A with nylon (n = 14) and group B with polyglycolic acid (n = 14). In each group, 8 animals had a vas deferens section and 6 had a previous vasectomy by ligature. Ten days postoperatively, the patency rate of the anastomosis was evaluated by the presence of sperm on both sides of the suture line. The contractility was assessed by mechanical stimulation. The existence of a sperm granuloma was considered as indicative of a non functional anastomosis. Three days later a fertility test was performed, lasting three months, and the number of litters was checked. The rats were sacrificed after three months, and each vas deferens was examined histologically or by electron microscope. The macroscopic results were: 57% patent anastomoses in group A and 77% in group B. 16% patent anastomoses after ligature in group A (n = 6) and 75% in group B (n = 6). The pregnancy rate was 54% in group A and 77% in group B. After previous ligation, the corresponding figures were 20% and 83% respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990320 TI - [Enkephalinergic receptors at the level of the lower urinary tract]. AB - After describing the autonomic nervous system of the lower urinary tract and its neurotransmitters, the authors discuss the recent discovery of new endomorphine group mediators--the enkephalins. There exist enkephalin immunoreactive nerve fibers in the smooth muscles of the bladder and the prostate. This neuromediator is synthetized in the spinal cord in the body of the preganglionic neuron and is carried by the axon flow towards the intramural parasympathetic ganglions of the bladder. The physiological effects of enkephalins are similar to those of morphinics and are antagonized by naloxone. Enkephalins inhibit spontaneous or provoked contractions of the bladder by inhibiting action on the parasympathetic neurons. This property of relaxing the muscles of the bladder, in both volume and pressure, may open up new vistas for uropharmacological research. PMID- 2990321 TI - [Cis-active function of plasmid R 57 resolving co-integrates formed during transposition of ISI-elements]. AB - The mechanism of pBR322 plasmid mobilization in the cells of Escherichia coli K 12 recA by conjugative factor R57 was studied. It was shown that mobilization of pB322 is achieved by formation of unstable IS1-mediated cointegrates with R57. In the rec+ E. coli strains cointegrates resolved with formation of pBR322:IS1 plasmids. In the recA bacteria the cointegrates dissociated to pBR:IS1, as well as to other insertion derivatives of pBR322. Some of the latter contain Tn9-like sequence, i.e. a transposon flanked by direct repeats of IS1. The subsequent transposition of IS1 from pBR-. IS1 to pBR3.1 plasmid (Aps deletion derivative of RP1) leads to formation of stable cointegrates incapable of dissociating even in the presence of coresident plasmid R57. It is suggested that R57 encodes the cys acting function providing recA-independent recombination between direct repeats of IS1. PMID- 2990322 TI - Inhibition of growth of Chlamydia trachomatis by nonoxynol-9 in vitro. AB - We evaluated the ability of the widely used spermicide nonoxynol-9, Conceptrol gel containing nonoxynol-9, and Conceptrol vehicle (without nonoxynol-9) to inhibit the formation of inclusions of Chlamydia trachomatis in cycloheximide treated McCoy cells. Conceptrol vehicle produced a non-dose-related 40 to 59% reduction of the number of inclusions formed. In contrast, the addition of nonoxynol-9 and Conceptrol gel containing nonoxynol-9 at a concentration of 100 micrograms/ml reduced the number of inclusions by 84 to 95%. Prolongation of contact between nonoxynol-9 and the chlamydial organisms from 90 to 180 min did not result in additional inhibition. Using a higher concentration of chlamydial organisms and diluting the drug-chlamydia mixture before inoculation of the monolayers, we were able to test higher concentrations of nonoxynol-9. At concentrations of 100 to 5,000 micrograms of nonoxynol-9 per ml, a dose-related 19 to 84% reduction of the number of inclusions was noted. PMID- 2990323 TI - Biotransformation and elimination of [2-14C]-1-(2-deoxy-2'-fluoro-beta-D arabinofuranosyl)-5-iodocytosine in immunosuppressed patients with herpesvirus infections. AB - The metabolism of the drug [2-14C]-1-(2'-deoxy-2'-fluoro-beta-D arabinofuranosyl)-5-iodocytosine (FIAC), a potent inhibitor of herpesvirus replication, was studied in immunosuppressed patients with herpesvirus infections. FIAC was administered intravenously by 15-min infusion and by mouth 24 h later to four patients at doses of 50 or 100 mg/m2. FIAC was cleared from the plasma primarily by biotransformation in liver, kidney, and peripheral blood, with a terminal-phase half-life of 0.92 to 1.80 h (mean, 1.36 h) after intravenous administration. The area under the concentration-time curve from zero to infinity (AUC0-infinity) for FIAC was 1.6 to 4.7% (mean, 3.4%) of the AUC0 infinity for total radioactivity. 1-(2'-Deoxy-2'-fluoro-beta-D-arabinofuranosyl) 5-iodouracil (FIAU) was the major metabolite; the AUC0-infinity for FIAU was 54.3 to 72.5% (mean, 63.4%) of the AUC0-infinity for total radioactivity. The terminal phase half-life for FIAU was 3.32 to 4.49 h (mean, 3.91 h); FIAU was cleared from plasma by renal elimination and further biotransformation. lesser amounts of 1 (2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)uracil, 1-(2'-deoxy-2'-fluoro-beta-D arabinofuranosyl)cytosine, the glucuronide conjugates of these metabolites, and the glucuronide conjugates of FIAC and FIAU were also formed. A comparison of the AUC0-infinity for total radioactivity after intravenous and oral administration suggested that nearly all of the oral dose was absorbed. Plasma levels of FIAU, also a potent inhibitor of herpesvirus replication in vitro, exceeded the 50% effective dose for herpes simplex virus and varicella-zoster virus as late as 12 h after administration of FIAC. PMID- 2990324 TI - Development of two DNA probes for differentiating the structural genes of subclasses I and II of the aminoglycoside-modifying enzyme 3'-aminoglycoside phosphotransferase. AB - Two DNA probes were developed to screen for the genes encoding 3'-aminoglycoside phosphotransferase activity in gram-negative bacilli. The 3'-I phosphotransferase [APH(3')I] probe was subcloned from Tn903; the APH(3')II probe was subcloned from Tn5. Each probe proved to be specific for genes corresponding to its own APH(3') subclass and did not hybridize with DNA from other classes when tested at high stringency by either Southern hybridization or dot-blot hybridization methods. The APH(3')I probe hybridized to DNA obtained from organisms demonstrating APH(3')I activity as measured by the phosphocellulose paper binding assay (PPBA) as well as to DNA from organisms reported to have both APH(3')I and APH(3')II activity by PPBA. This probe did not hybridize to DNA from organisms showing only APH(3')II activity by PPBA. The APH(3')II probe demonstrated homology with DNA from organisms showing APH(3')II activity by PPBA but not with DNA from organisms showing APH(3')I activity or both APH(3')I and APH(3')II activity by PPBA. We conclude that organisms previously believed to contain both APH(3')I and APH(3')II genes based on PPBA contain in fact only the APH(3')I gene. PMID- 2990326 TI - In vitro activity of the spermicide nonoxynol-9 against Chlamydia trachomatis. AB - The in vitro activity of nonoxynol-9 against four serotypes (C, D, H, and K) of Chlamydia trachomatis was investigated. A constant inoculum of each serotype was exposed to serial twofold dilutions (1:100 to 1:800) of Koromex, Conceptrol, or reference preparations (not containing nonoxynol-9) for 4 and 24 h at 37 degrees C. The mixtures of nonoxynal-9 or nonnonoxynol preparations and control inocula were dispensed into triplicate wells containing McCoy cell monolayers. After incubation at 37 degrees C, the monolayers were fixed and stained with iodine and examined for evidence of infection with C. trachomatis. All nonoxynol-9 containing preparations showed marked antichlamydial activity as judged by percent reduction of glycogen-containing intracytoplasmic inclusions. The reference preparations, which did not contain nonoxynol-9, were markedly less active when tested in this in vitro system. PMID- 2990325 TI - Antiherpes effects and pharmacokinetic properties of 9-(4-hydroxybutyl) guanine and the (R) and (S) enantiomers of 9-(3,4-dihydroxybutyl)guanine. AB - Three acyclic guanosine analogs with similar structures, the (R) and (S) forms of 9-(3,4-dihydroxybutyl)guanine and 9-(4-hydroxybutyl)guanine, were compared for antiherpes activity in vivo and in vitro. The three guanosine analogs were viral thymidine kinase-dependent inhibitors of virus multiplication. In cell cultures, (S)-9-(3,4-dihydroxybutyl)guanine was the least active of these three drugs against a variety of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) strains. This was also the case for a certain HSV-1 or HSV-2 strain in different cell lines. In cell cultures, (R)-9-(3,4-dihydroxybutyl)guanine and 9-(4 hydroxybutyl)guanine had similar antiherpes activities. However, in vivo in cutaneous HSV-1 infections in guinea pigs treated topically and in systemic HSV-2 infections in mice treated orally or intraperitoneally, only (R)-9-(3,4 dihydroxybutyl)guanine had a therapeutic effect. The extremely short half-life in plasma and the high clearance of 9-(4-hydroxybutyl)guanine as compared with those of (R)-9-(3,4-dihydroxybutyl)guanine probably made 9-(4-hydroxybutyl)guanine inefficacious when given intraperitoneally or orally to mice infected with herpesvirus. On the other hand, no kinetic differences between (R)-9-(3,4 dihydroxybutyl)guanine and 9-(4-hydroxybutyl)guanine were observed in penetration through guinea pig skin ex vivo, and no preferential metabolism of 9-(4 hydroxybutyl)guanine in skin was noted. We deduced that high thymidine levels in guinea pig skin preferentially antagonize the antiviral effect of 9-(4 hydroxybutyl) guanine in cutaneous HSV-1 infections. PMID- 2990327 TI - In vitro susceptibilities of 126 clinical isolates of Yersinia enterocolitica to 21 beta-lactam antibiotics. AB - The MICs of 21 beta-lactam antibiotics were measured against 126 clinically significant and epidemiologically unrelated Yersinia enterocolitica isolates. The most active antimicrobial agents tested (geometric means of MICs) were ceftriaxone, cefotaxime, ceftizoxime, and cefmenoxime (0.06 to 0.08 micrograms/ml). Mezlocillin (1.36 micrograms/ml) and piperacillin (1.57 micrograms/ml) were the most active penicillins. Aztreonam and imipenem had MICs of 0.44 and 0.24 micrograms/ml. All isolates exhibited some degree of resistance against ampicillin (MICs, greater than or equal to 4 micrograms/ml) and cephalothin (MICs, greater than or equal to 8 micrograms/ml). Cephalosporinase or penicillinase activities were expressed by all isolates. A principal component analysis of MIC data separated the major serotypes (O3, O9, O8, and O5,27). The MCBs of 14 newer beta-lactams were measured against 10 clinical isolates. On the basis of the ratio of MBC to MIC (expressed in dilution factors), a real bactericidal activity was only observed for imipenem (MBC/MIC = 1). For the other newer beta-lactams, the ratios ranged from 4 to 8. PMID- 2990328 TI - Activity of UK-49,858, a bis-triazole derivative, against experimental infections with Candida albicans and Trichophyton mentagrophytes. AB - The therapeutic potential of UK-49,858, a difluorophenyl bis-triazole derivative, has been assessed by evaluating its activity against systemic infections with Candida albicans in normal mice and rats and in mice with impaired defence mechanisms, against vaginal infections with C. albicans in mice, and against dermal infections with Trichophyton mentagrophytes in guinea pigs. Orally administered ketoconazole was used as a comparative agent throughout, and parenterally administered amphotericin B was included in the study of C. albicans systemic infection in normal mice. The activity of UK-49,858 given orally to mice or rats infected systemically with C. albicans was far superior to that of ketoconazole. In addition, UK-49,858 showed activity comparable to that of amphotericin B when given parenterally, although the latter gave more prolonged protection. UK-49,858 was also effective orally in curing experimental candidal vaginitis in mice and trichophytosis in guinea pigs, against which it was approximately 10 times more active than ketoconazole. These data suggest that UK 49,858 may be of value in the treatment of both C. albicans and dermatophyte fungal infections in man. PMID- 2990329 TI - Studies on 44 081 R.P., a new antirhinovirus compound, in cell cultures and in volunteers. AB - A synthetic compound, 2-[(1,5,10,10a-tetrahydro-3H-thiazolo[3,4b]isoquinolin-3 ylidene) amino]-4-thiazoleacetic acid (S), 44 081 R.P., inhibits the multiplication of rhinoviruses in cell cultures. Of the 69 rhinovirus strains and serotypes that have been studied, 39% were inhibited at a concentration of 7 micrograms/ml, far below that which affects cellular metabolism (250 micrograms/ml). Preliminary data indicate that the compound inhibits some early events of virus replication but that some cellular functions are also involved in its mechanism of action. Despite its antiviral activity in vitro, the compound, when self-administered intranasally as a 0.2% solution to volunteers from the day before to 5 days after inoculation with a human rhinovirus strain, had no significant effect on rhinorrhea, clinical score, or laboratory evidence of infection. PMID- 2990330 TI - In vitro efficacy of sulbactam combined with ampicillin against anaerobic bacteria. AB - An ampicillin-sulbactam combination was compared with ampicillin alone, chloramphenicol, clindamycin, and metronidazole against 272 strains of anaerobic bacteria. Chloramphenicol and ampicillin-sulbactam were the most effective, inhibiting 98 to 99% of strains tested at breakpoint (16 micrograms/ml). The combination of sulbactam and ampicillin was much more effective than ampicillin alone against Bacteroides fragilis strains but did not differ substantially from ampicillin alone against Fusobacterium spp., gram-positive rods, and gram positive cocci. PMID- 2990331 TI - Resistance to various tetracyclines mediated by transposon Tn10 in Escherichia coli K-12. AB - Levels of resistance to tetracycline, chlortetracycline, demethylchlortetracycline, doxycycline, oxytetracycline, methacycline, pyrrolidinotetracycline, minocycline, and beta-chelocardin of Escherichia coli K 12 carrying transposon Tn10 or defined DNA segments of Tn10 were determined. In all cases, tetA was the only gene required for resistance. Doxycycline was the most effective inducer of tetA gene expression. PMID- 2990332 TI - Rhinovirus inactivation by nasal tissues treated with virucide. AB - Rhinovirus colds may be transmitted by hand-to-hand contact followed by self inoculation of nasal and/or conjunctival mucosa with virus contaminating the fingertips. The purpose of this study was to determine whether impregnation of nasal tissues with virucidal compounds could prevent rhinovirus from passing through the tissue and thus provide a means of preventing hand contamination during nose blowing. Paper tissues treated with a combination of citric acid, malic acid, and sodium laruryl sulfate were compared to placebo tissues containing sodium saccharin. Recovery of infectious virus was significantly reduced by passage of the virus-containing medium through virucidal versus placebo tissue (1/18 vs. 17/18 respectively, P less than 0.001, Fisher exact test). The virucidal effect of treated tissues was demonstrated for multiple rhinovirus serotypes suspended in either cell culture medium or nasal mucus. Virus contained in mucus from infected volunteers was also inactivated. PMID- 2990333 TI - Thio-analogues of inorganic pyrophosphate inhibit the replication of influenza virus A in vitro. AB - Mono- and bisthiopyrophosphate can inhibit the replication of influenza virus A/X49 in Madin-Darby canine kidney (MDCK) cells at concentrations at which no cytotoxic effect is observed after 3 days. The thiopyrophosphate analogues inhibit the RNA transcriptase activity of this virus possibly by chelating with an essential metal ion in the transcriptase complex. [31P]NMR spectroscopy indicates that bisthiopyrophosphate coordinates to zinc through sulphur and magnesium through oxygen which may influence the inhibitory properties of this compound with metal-containing enzymes. PMID- 2990334 TI - Effects of combined use of acyclovir and antibody in athymic nude mice inoculated intracutaneously with herpes simplex virus. AB - Antiviral effects of acyclovir (ACV) and antibody were studied in athymic nude mice inoculated intracutaneously in the midflank with herpes simplex virus type 1. Three hours after virus inoculation, treatment was initiated. In ACV-treated mice, the development of skin lesions was inhibited and the mean survival time was prolonged as compared with controls. Treatment with ACV markedly reduced the viral titers both at the inoculation site and in the neural tissues (dorsal root ganglia, spinal cord and brain). Transfer of antibody was also effective in controlling infection as described previously. The use of ACV plus antibody was more effective than the use of ACV alone and in 5 of 17 mice the disease was completely inhibited. Furthermore, in the mice that survived, latent infections in the ganglia were also prevented. These results indicate that treatment with ACV plus antibody is highly effective against herpes simplex virus infection in the immunocompromised host. PMID- 2990335 TI - Molecular assessment of S1 endonuclease-resistant snapback hairpin loops generated by DNA polymerase I during the in-vitro nick translation reaction. AB - The in-vitro nick translation reaction used to label DNA to high specific activity also produces aberrant DNA structures known as "snapback" hairpin loops. Hairpin structures are precluded from participating in precise DNA-DNA hybridization interactions. Three nick translation systems were all found to yield significant quantities of snapback hairpins, as determined by their resistance to S1 endonuclease digestion following denaturation. The relative quantities of hairpins produced correlated with both the mass average size of the final DNA probe product synthesized as well as the overall rate of the nick translation reaction. Decreases in the amount of exogenous DNase I used in nick translation reactions produced significant decreases in the amount of hairpin loop structures formed. Hairpins could be effectively removed from nick translated DNAs by employing hydroxylapatite column chromatography. Strategies to reduce hairpin formation during nick translation and the removal of hairpins from nick-translated DNAs are presented. PMID- 2990336 TI - UV inactivation of pathogenic and indicator microorganisms. AB - Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts. PMID- 2990337 TI - Elimination of viruses and indicator bacteria at each step of treatment during preparation of drinking water at seven water treatment plants. AB - Seven drinking water treatment plants were sampled twice a month for 12 months to evaluate the removal of indicator bacteria and cytopathogenic enteric viruses. Samples were obtained at each level of treatment: raw water, postchlorination, postsedimentation, postfiltration, postozonation, and finished (tap) water. Raw water quality was usually poor, with total coliform counts exceeding 105 to 106 CFU/liter and the average virus count in raw water of 3.3 most probable number of cytopathogenic units (MPNCU)/liter; several samples contained more than 100 MPNCU/liter. All plants distributed finished water that was essentially free of indicator bacteria as judged by analysis of 1 liter for total coliforms, fecal coliforms, fecal streptococci, coagulase-positive staphylococci, and Pseudomonas aeruginosa. The total plate counts at 20 and 35 degrees C were also evaluated as a measure of the total microbial population and were usually very low. Viruses were detected in 7% (11 of 155) of the finished water samples (1,000 liters) at an average density of 0.0006 MPNCU/liter the highest virus density measured being 0.2 MPNCU/liter. The average cumulative virus reduction was 95.15% after sedimentation and 99.97% after filtration and did not significantly decrease after ozonation or final chlorination. The viruses isolated from treated waters were all enteroviruses: poliovirus types 1, 2, and 3, coxsackievirus types B3, B4, and B5, echovirus type 7, and untyped picornaviruses. PMID- 2990338 TI - The aggregation state of bovine heart cytochrome c oxidase and its kinetics in monomeric and dimeric form. AB - The monomeric and dimeric forms of bovine cytochrome c oxidase (EC 1.9.3.1) were obtained from gel filtration chromatography on Ultrogel AcA 34 and analyzed. Both species contained all 12-13 subunits described for this enzyme. In the dimer 320 molecules [3H]dodecyl-beta-D-maltoside were bound per heme aa3 and in the monomer 360 molecules per heme aa3. The monomers contained 10 mol of tightly bound phospholipid/mol heme aa3 and the dimers 14. Sedimentation coefficients of 15.5 18 S for the dimer and 9.6 S for the monomer were calculated from sucrose density centrifugation analysis and analytical centrifugation. By the laser beam light scattering technique a Stokes radius of 70 A for the dimeric detergent-lipid protein complex was measured. From those parameters and the densitometric determined partial specific volumes of the detergent and the enzyme, the molecular weights of 400,000 for the protein moiety of the dimer and 170,000 200,000 for the monomer were calculated. Under very low ionic strength conditions the monomer/dimer equilibrium was found to be dependent on the protein concentration. At low enzyme concentrations (10(-9) M) monomers were predominant, whereas at concentrations above 5 X 10(-6) M the amounts of dimers and higher aggregates were more represented. The cytochrome c oxidase activity, measured spectrophotometrically and analyzed by Eadie-Hofstee plot, was biphasic as a function of cytochrome c concentration for the dimeric enzyme. Pure monomers gave monophasic kinetics. The data, fitting with a homotropic negative cooperative mechanism for the dimer of cytochrome c oxidase, are discussed and compared with other described mechanisms. PMID- 2990339 TI - The effect of concentration on the antioxidant effectiveness of alpha-tocopherol in lipid peroxidation induced by superoxide free radicals. AB - Egg yolk phosphatidylcholine liposomes were rapidly oxidized in the presence of chelated iron and a superoxide-generating system. alpha-Tocopherol incorporated in the bilayer was oxidized at the same time. No lipid or alpha-tocopherol oxidation occurred in liposomes composed of dimyristoyl phosphatidylcholine. The antioxidant did not inhibit lipid peroxidation until its concentration reached a critical level, which depended on the effectiveness of the oxidative stress. Beyond this level, peroxidation was inhibited completely and, simultaneously, the rate of oxidation of tocopherol was lowered. The results suggest that the antioxidant efficiency of alpha-tocopherol depends on its ability to react mainly with the chain-initiating or chain-propagating lipid radicals. This, in turn, is closely tied to the tocopherol content of the membrane. Ascorbate inhibited the consumption of alpha-tocopherol, possibly by regenerating its reduced form. PMID- 2990340 TI - Inhibition and activation of bovine heart NAD-specific isocitrate dehydrogenase by ATP. AB - In the absence of added calcium, inhibition of NAD-specific isocitrate dehydrogenase by ATP occurred without ADP (I0.5 = 1.8 mM) and with 0.2 mM ADP3- (I0.5 = 1.0 mM) at subsaturating substrate concentrations at pH 7.4. Inhibition by ATP was competitive with NAD+ in the presence and absence of ADP and was not reversed by magnesium citrate. No reversal of ATP inhibition by free Ca2+ was observed in the presence of ADP (0.2 mM). However, when ADP was absent, increasing Ca2+ first caused progressive reversal of ATP inhibition followed by activation by ATP. Without ADP, the S0.5 for calcium activation was 80-140 microM at ATP concentrations between 0.6 and 3.0 mM. The S0.5 for ATP activation, in the absence of ADP, was 1.1 and 2.1 microM when free Ca2+ was held constant at 0.1 and 1.0 mM, respectively. As in activation by ADP, ATP decreased the S0.5 for magnesium isocitrate without affecting V. However, in contrast to ADP, the activation by ATP occurred without lowering the Hill coefficient for the substrate. GDP activated the enzyme at relatively high concentrations of Ca2+ but not without added Ca2+. PMID- 2990341 TI - A major phosphotyrosyl-protein phosphatase from bovine heart is associated with a low-molecular-weight acid phosphatase. AB - The phosphotyrosyl [Tyr(P)]-immunoglobulin G (IgG) phosphatase activity in the extracts of bovine heart, bovine brain, human kidney, and rabbit liver can be separated by DEAE-cellulose at neutral pH into two fractions. The unbound fraction exhibits a higher activity at acidic than neutral pH while the reverse is true for the bound fraction. Of all tissues examined, the Tyr(P)-IgG phosphatase activity in the unbound fraction measured at pH 5.0 is higher than that in the bound fraction measured at pH 7.2. The acid Tyr(P)-IgG phosphatase activity has been extensively purified from bovine heart. It copurified with an acid phosphatase activity (p-nitrophenyl phosphate (PNPP) as a substrate) throughout the purification procedure. These two activities coelute from various ion-exchange and gel filtration chromatographies and comigrate on polyacrylamide gel electrophoresis, indicating that they reside on the same protein molecule. The phosphatase has a Mr = 15,000 by gel filtration and exhibits an optimum between pH 5.0 and 6.0 when either Tyr(P)-IgG-casein or PNPP is the substrate. It is highly specific for Tyr(P)-protein with little activities toward phosphoseryl [Ser(P)]- or phosphothreonyl [Thr(P)]-protein. The enzyme activities toward Tyr(P)-casein and PNPP are strongly inhibited by microM molybdate and vanadate but insensitive to inhibition by L(+)-tartrate, NaF, or Zn2+. The molecular and catalytic properties of the acid Tyr(P)-protein phosphatase purified from bovine heart are very similar to those of the low-molecular-weight acid phosphatases of Mr = 14,000 previously identified and purified from the cytosolic fraction of human liver, placenta, and other animal tissues. PMID- 2990342 TI - Identification of a 19-kDa polypeptide as an Fe-S center apoprotein in the photosystem I primary electron acceptor complex. AB - Treatment of Photosystem I (PSI) with sodium thiocyanate, a chaotropic agent, results in the selective depletion of certain low-molecular-weight polypeptides. A PSI complex obtained following treatment with 0.5 M sodium thiocyanate is significantly depleted of polypeptides of approximately 8, 10, 14, and 16 kDa, relative to an untreated control, but retains approximately 90% of the EPR signal amplitude associated with the iron-sulfur Centers A and B. The only peptides remaining that could not be depleted without a parallel decrease in the signal amplitude of the Fe-S Centers A and B are the 62-kDa reaction center-containing polypeptide and a 19-kDa polypeptide. These results are considered in relation to the identity of the apoprotein of the Fe-S Centers A and B. PMID- 2990343 TI - Interactions of glucocorticoids with the AtT-20 cell: effect on protein accumulation. AB - In order to investigate the mechanism through which glucocorticoids downregulate the number of their own receptors in the AtT-20 cell, the effect of glucocorticoids on cell protein metabolism was studied. Glucocorticoids were found to inhibit cellular protein accumulation when included in long-term cultures. The concentrations of agonists that cause a mid-maximal effect are similar to those needed to half-saturate the glucocorticoid receptor, suggesting that the growth-inhibiting effect is receptor-mediated. Two-dimensional electrophoresis of cytosolic extracts of treated and control cells suggested that the effect reflected a general suppression of overall protein accumulation rather than a selective effect on certain classes. Comparison of the protein to DNA ratio of control and dexamethasone-treated cells showed that the latter have higher ratios suggesting that cell composition may be altered by agonists. However, time-course studies of this effect indicated that this is basically an expression of a glucocorticoid effect on cell growth rather than a selective effect on protein metabolism. It is concluded that glucocorticoids inhibit overall AtT-20 cell growth and that this, in turn, manifests itself as a decrease in the rate of protein accumulation. It is suggested that this change in protein metabolism may be a minor component in the mechanism through which glucocorticoids decrease AtT-20 cell ACTH secretion and glucocorticoid receptor number. PMID- 2990344 TI - Changes of intestinal alkaline phosphatase produced by cholecalciferol or 1,25 dihydroxyvitamin D3 in vitamin D-deficient chicks. AB - Treatment with cholecalciferol or 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) increases activity and changes electrophoretic mobility of alkaline phosphatase (alkPase) from duodenal brush border of vitamin D-deprived chicks. Three of the four molecular forms of the enzyme show reduced velocity of migration 9 h after 1,25(OH)2D3 or 24 h after vitamin D3. This change is reversed about 48 h later, when mobility of those bands is higher than that of controls. Incubation of enzyme preparations with exogenous neuraminidase produces the same electrophoretic modifications observed during the early stage, indicating that they are due to desialylation. Cholecalciferol or 1,25(OH)2D3 increase sialidase activity of duodenal brush border. This increment precedes that of alkPase and could account for the initial desialylation and moderate rise of alkPase. Cycloheximide markedly reduces alkPase in rachitic chicks and blocks the increase of the enzyme activity produced by vitamin D3, but does not modify the rise of sialidase or the reduction of alkPase electrophoretic mobility. The bimodal response of alkPase to 1,25(OH)2D3 or cholecalciferol comprises two different mechanisms: during a first stage, epigenetic modifications of preexisting enzyme can be triggered by the increased Ca2+ levels; in a second phase, there is activation of enzyme synthesis. PMID- 2990345 TI - Biosynthesis rates and content of thymosin beta 4 in cell lines. AB - The content and relative biosynthetic rates of thymosin beta 4 have been determined in 28 different cell lines. The highest content of thymosin beta 4 as well as the highest rate of biosynthesis was observed in Epstein-Barr virus transformed human B-cell lines. The levels observed in these cells are 1 pg thymosin beta 4 per cell, which is three times higher than that in rat peritoneal macrophages. Thus, these B-cell lines have the highest content of thymosin beta 4 of any cell type yet described. Since all of the Epstein-Barr virus-transformed B cells described here grow in suspension, it is unlikely that the presence of thymosin beta 4 is related to anchorage in these cells. Thymosin beta 4 is not secreted by viable Epstein-Barr virus-transformed B cells in culture, suggesting some intracellular function of the peptide. These results indicate that these B cell lines may be suitable for the study of thymosin beta 4 function. PMID- 2990346 TI - Enkephalin-degrading dipeptidyl carboxypeptidase in guinea pig serum: its properties and action on bioactive peptides. AB - A dipeptidyl carboxypeptidase, which cleaved the Gly3-Phe4 bond of enkephalins, was purified from guinea pig serum 420-fold. The optimum pH of the enzyme was in the neutral range (pH 7.25), and the molecular weight was estimated to be approx. 280,000. The enzyme hydrolyzed Met- and Leu-enkephalin with Km values of 0.30 and 0.50 mM, respectively. The enzyme was inhibited by metal chelators and p-chloro mercuribenzoate. Captopril showed high inhibitory potency, while phosphoramidon and Phe-Ala showed no effect on the enzyme activity. Therefore, the obtained enzyme can be classified as an angiotensin-converting enzyme (EC 3.4.15.1). Among the bioactive peptides examined, bradykinin and angiotensin I were hydrolyzed by the enzyme. Angiotensin III showed a stronger inhibitory effect than that of angiotensin II. Substance P, gastrin I, and secretin were also inhibitory toward the enzyme activity. On high-performance liquid chromatography analysis, Met enkephalin-Arg6-Phe7 and Leu-enkephalin-Arg6 were cleaved sequentially at the second peptide bond of the C terminus. Thus, the dipeptidyl carboxypeptidase in guinea pig serum may play a role not only in the angiotensin-bradykinin system but also in the metabolism of circulating enkephalins and other bioactive peptides. PMID- 2990347 TI - Characterization of protein transport between successive compartments of the Golgi apparatus: asymmetric properties of donor and acceptor activities in a cell free system. AB - Transport of the vesicular stomatitis virus (VSV)-encoded glycoprotein (G protein) between successive compartments of the Golgi in a cell-free system is measured by the coupled incorporation of N-[3H]acetylglucosamine (GlcNAc). This glycosylation occurs when G protein is transported from a "donor" compartment in Golgi membranes that lack GlcNAc transferase I (from VSV-infected CHO clone 15B cells) to the next "acceptor" compartment in a Golgi population from wild-type CHO cells (containing the GlcNAc transferase but not G protein). Here we present a detailed characterization of the conditions required to achieve transport in vitro. We find that donor and acceptor activities differ markedly in certain of their properties. The donor activity is inhibited by N-ethylmaleimide but the acceptor activity is resistant. Donor activity is unstable in the absence of ATP or the cytosol fraction; acceptor activity is much more stable. This asymmetry may reflect the vectorial nature of the underlying biochemistry of protein transport. Both donor and acceptor are trypsin-sensitive, implying a need for cytoplasmically oriented membrane proteins. Transport occurs only in a restricted range of close to physiological conditions. ATP is absolutely required, although as little as 1 microM is sufficient. Transport is inhibited by ATP-gamma-sulfate and vanadate, suggesting that ATP hydrolysis is needed. By contrast, ionophores that dissipate membrane potentials and proton gradients do not inhibit transport. Monensin was also without effect in the cell-free system. PMID- 2990348 TI - S-100 protein-induced changes in the physical state of synaptosomal particulate fractions as monitored by spin labels. AB - This report documents changes in the physical state of synaptosomal particulate fractions (SYN) upon binding of S-100 protein, as monitored by spin labels. Studies were conducted on SYN labeled with either 5-doxylstearic acid or 16 doxylstearic acid, which probe the polar region and the hydrophobic core of the lipid bilayer, respectively. S-100 perturbs to some extent both the polar surface and the hydrophobic core of SYN in a time- and temperature-dependent manner. Ca2+ is essential for S-100 to perturb the membranes. K+ almost completely inhibits the S-100 perturbing effect if present in the incubation medium, but fails to reverse the S-100-induced changes if added after S-100 has interacted with SYN. At room temperature and below, the overall S-100 effect registered after about 30 min of association of the protein with SYN is an increase in the fluidity of both the surface and the interior of the membranes. Spectra registered at intervals at room temperature indicate that the S-100 perturbing effect on the membrane surface is practically monophasic, consisting of an increase in fluidity, while that on the membrane interior is multiphasic, consisting of a decrease in fluidity during the first 10 min of association, followed by an increase in fluidity during the subsequent 20 min and a return to starting values during the second 30 min of association. Around 37 degrees C, on the contrary, a decrease in fluidity is registered in both regions. The data suggest that S-100 induces a spatial rearrangement of membrane components (proteins) involved in the specific binding and/or partially penetrates into the lipid bilayer. PMID- 2990349 TI - High-pressure investigations of cytochrome P-450 spin and substrate binding equilibria. AB - The effects of high pressure (1-2000 bar) on the spin state and substrate binding equilibria in cytochrome P-450 have been determined. The high-spin (S = 5/2) to low spin (S = 1/2) transition of the ferric hemoprotein was monitored by uv visible spectroscopy at various substrate concentrations. Increasing hydrostatic pressure on a sample of substrate-bound cytochrome P-450 resulted in a decrease in the high-spin fraction as monitored by a Soret maxima at 391 nm and an increase in the low-spin 417-nm region of the spectrum. These pressure-induced optical changes were totally reversible for all pressures below 800 bar and were found to correspond to simple substrate dissociation from the enzyme. High levels of the normally metabolized substrate, d-camphor, corresponding to a 99.9% saturation of the hemoprotein active site (50 mM Tris-Cl, 100 mM KCl, pH 7.2) completely prevented the pressure-induced high-spin to low-spin transition that is observed at less than saturating substrate concentrations. A gradual increase in the formation of the inactive P-420 form of the cytochrome was noted if the pressure of the sample was increased above 800 bar. These pressure-linked spectral changes were used to determine the microscopic volume change accompanying substrate binding, which was found to be -47.0 +/- 2 ml/mol (pH 7.2) which represents a substantial change for a ligand dissociation reaction. The observed volume change for camphor binding decreases to -30.6 +/- 2 ml/mol at pH 6.0, suggesting the involvement of a linked proton equilibrium. Various substrate analogs of camphor induce varying degrees of low-spin to high-spin shift upon binding to ferric cytochrome P-450 (3). The volume changes for the dissociation of these substrates were very similar to those obtained with camphor. The conformational changes associated with a shift from high- to low-spin ferric iron appear to be small in comparison to the overall macroscopic changes in volume accompanying substrate binding to the enzyme. PMID- 2990350 TI - [Clinical trial of MCNU for malignant brain tumors]. AB - A total of 71 cases with primary brain tumors (44 cases) and metastatic brain tumors (30 cases) were entered into our clinical studies with MCNU and radiation therapy or MCNU alone. With regard to tumor reduction on CT scan, the response rates obtained for MCNU and radiation were 21.7% for malignant gliomas and 50.0% for metastatic brain tumors. With regard to improvement of neurological signs, the response rates obtained for MCNU and radiation were 62.9% for malignant gliomas and 71.4% for metastatic brain tumors. The response rates for MCNU were 5.8% for malignant gliomas and 46.1% for metastatic brain tumors. In the improvement of performance status, the response rates for MCNU and radiation were 51.8% for malignant gliomas and 64.2% for metastatic brain tumors. The response rates for MCNU were 46.1% for malignant gliomas and 30.7% for metastatic brain tumors. A minimal degree of hematological toxicity occurred but this gradually disappeared. These results suggested that MCNU has relatively effective antitumor activity against metastatic brain tumors and an enhanced effect with radiation against malignant gliomas. PMID- 2990351 TI - [cis-Dichlorodiammine platinum (II) in the treatment of advanced colo-rectal cancer]. AB - The effect of CDDP was evaluated in 10 cases of advanced colo-rectal cancer. Prior chemotherapy was done in seven cases and three were fresh cases. The dosage of CDDP was in the range of 70-100 mg/m2 per individual. Seven cases out of 10 receiving the CDDP chemotherapy achieved no change and there were three cases of PD. Response rate was 0%. Values of serum creatinine and BUN were transiently evaluated and bone marrow toxicity was moderate. Emesis of patients treated with CDDP was suppressed by the administration of high doses of metoclopramide (2 mg/kg X 4 times). PMID- 2990352 TI - [Effect of adriamycin, mitomycin-C, and tegafur (AMF) chemotherapy on advanced lung cancer]. AB - Seven patients with advanced inoperable carcinoma of the lung were treated with combination chemotherapy consisting of adriamycin, mitomycin-C, and tegafur (AMF therapy). There was, at least, no progress of the carcinoma during treatment and objective response was obtained in 28.6% of patients. With regard to side effects, leukopenia and gastrointestinal symptoms were found in each of 3 patients. PMID- 2990353 TI - Bowenoid papulosis. Presence of human papillomavirus (HPV) structural antigens and of HPV 16-related DNA sequences. AB - This study reviews 39 cases of anogenital bowenoid papulosis lesions in 22 individuals of both sexes that were analyzed clinically, histologically, immunocytochemically, and virologically. Macroscopically, three different types of lesions were demonstrated: erythematous macules; papules (lichenoid and/or pigmented papules); and leukoplakialike lesions. Microscopically, bowenoid papulosis fulfills the criteria of a squamous cell carcinoma in situ. Much like oral precancers, three distinct growth patterns (flat, endophytic, and exophytic) could be differentiated, which did not correlate with the clinical aspect of the lesions. In only two (5.12%) of the 39 cases of bowenoid papulosis could structural antigens of papillomaviruses be detected immunocytochemically (peroxidase-antiperoxidase technique). The DNA from 12 lesions that were analyzed for the presence of papillomavirus-specific sequences hybridized stringently in all cases with the human papillomavirus 16 specific DNA probe labeled with phosphorus 32. PMID- 2990354 TI - X-linked inheritance of epidermodysplasia verruciformis. Genetic and virologic studies of a kindred. AB - We describe a family with typical epidermodysplasia verruciformis (EV) in which only male members are affected. Whereas none of the index patient's ten children have EV, four of eight grandsons born to his daughters have inherited the disorder. All are infected with human papillomavirus (HPV) 3 and HPV 8. The inheritance in this kindred most likely results from an X-linked recessive genetic defect. Since other kindreds have been described with autosomal inheritance, this novel inheritance pattern suggests that the persistent high clinical susceptibility to HPV infection characteristic of EV may result from defects in either of at least two different genetic loci, one of which may be located on the X chromosome. PMID- 2990355 TI - Arm tumor resembling a supernumerary nipple. PMID- 2990356 TI - Long term prognosis for babies with neonatal liver disease. AB - A total of 123 patients with neonatal liver disease without extrahepatic bile duct obstruction or arteriohepatic dysplasia have been studied for six to 18 years. Idiopathic neonatal hepatitis, present in 73 babies, carried a high mortality due to liver failure (18%), septicaemia (6%), and associated defects (14%), especially in the first year of life (25%). Progression to chronic liver disease in non-familial idiopathic cases occurred in three of 40 reviewed patients. Only 12 of these children were completely healthy, the remainder having other permanent disabilities (57%). Four of nine familial cases of idiopathic neonatal hepatitis died in the first 12 months of life as did two of the four reviewed survivors. Progression to chronic liver disease or to death was a continuous process without any interval of recovery in all but one of these patients. Among patients with a presumed infective cause, cytomegalovirus infection caused a particularly benign form of neonatal hepatitis but was a frequent cause of brain damage or other disabilities. Babies who survived other infective liver diseases showed complete healing of the liver damage. Neonatal liver disease associated with alpha 1 antitrypsin deficiency progressed to death or chronic liver disease in three of nine patients and was not associated with a paucity of interlobular bile ducts. PMID- 2990357 TI - Filamentous aggregates of collagen. Ultrastructural evidence for collagen-fibril degradation in situ. AB - Filamentous aggregates of collagen are distinct structures in the pathological dermis. These aggregates are distinguishable from fibrous long-spacing collagen (in vitro and at biopsy) and the Luse body. The aggregates are produced from dermal collagen fibrils by clostridial collagenase and culture-medium extract, which supposedly contains cellular collagenase at a neutral pH, as well as by organ cultures. In vitro experiments showed that carrageenan granuloma contains fibrous long-spacing collagen and segment long-spacing collagen. The granuloma also contains the aggregates. The aggregates were found in skin biopsies from syphilitic chancres, acrosclerotic scleroderma, morphea, mycosis fungoides, myeloid leukemia, mastocytosis and malignant melanoma. These findings indicate that the aggregates are products of the in situ degradation of collagen fibrils by some collagenolytic factor. This factor may originate in fibroblast-like cells, reticulum cells, leukemia cells, mast cells and melanoma cells. PMID- 2990358 TI - Cystosarcoma phyllodes: epidemiology, pathohistology, pathobiology, diagnosis, therapy, and survival. AB - Cystosarcoma phyllodes is a very rare tumor which maybe difficult to diagnose clinically. The epidemiology and pathobiology are different from those of breast carcinoma. Risk factors, multicentricity, bilaterality, as associated with breast carcinoma, are not observed in patients with cystosarcoma phyllodes. Although the term "sarcoma" indicates a malignant tumor, only 10%-30% of cystosarcomas are histologically diagnosed as malignant; clinical diagnosis of malignancy does not exceed 10%. Axillary node involvement is rare, but hematogenous spread of cystosarcoma occurs into lung, pleura, bone, and liver. Clinically, cystosarcoma is a large (usually 3-5 cm in diameter) painless tumor with sudden growth acceleration especially during pregnancy. Cystosarcoma is usually circumscribed, containing firm and soft areas. The differential diagnosis has to include fibroadenoma, fibrocystic disease, mastitis, abscess, and medullary carcinoma. Neither clinical, mammographic or sonographic signs exist to predict a benign or malignant tumor. Therapy of cystosarcoma is not uniformly agreed upon. Radical, modified-radical, and simple mastectomy and tumorectomy are typical treatments; therapeutic results are the same for each treatment modality. For histologically diagnosed malignant cystosarcoma, the relative 5-year survival rate is about 80%. Clinically, malignant metastatic cystosarcoma is incurable; radiotherapy, endocrine treatment, and polychemotherapy are all ineffective. Because of the specific tumor pathobiology of cystosarcoma and its rarity, evaluation of treatment modalities and comparison of survival rates are difficult. PMID- 2990359 TI - One hundred hepatic resections. Indications and operative results. AB - The indications and operative results of hepatic resections were investigated in 100 consecutive patients over the past 20 years. There were 61 hepatocellular carcinoma, 13 hepatolithiasis, and 26 other miscellaneous diseases. An overall hospital mortality rate was 25%. It was 26, 0, and 35% in patients with hepatocellular carcinoma, hepatolithiasis, and other diseases, respectively. There was not any significant difference between survived and deceased cases in their preoperative laboratory data. The volume of operative blood loss in deceased cases was significantly larger than that in survived cases, so the influence of operative blood loss on morbidity and mortality was investigated. The incidences of postoperative bleeding, hepatic insufficiency, pulmonary insufficiency, and hospital death were significantly higher in patients whose operative blood loss exceeded 5000 ml. These results indicate that operative blood loss is one of the critical factors that decide the operative prognosis. PMID- 2990360 TI - The prevalence of appendiceal fecaliths in patients with and without appendicitis. A comparative study from Canada and South Africa. AB - Appendicitis is more common in developed than in developing societies and appendiceal fecaliths are thought to have an etiologic role in the disease. The geographic distribution of appendiceal fecaliths was investigated by systematic, intraoperative palpation of the appendix in patients in Toronto, Canada and Johannesburg, South Africa. The incidences of fecaliths found on pathologic sectioning of the appendix in appendicitis patients in both societies were compared. In the Canadian population, the prevalence of fecaliths in patients whose appendices were palpated incidentally was 32% versus 52% for those with appendicitis (p less than 0.01). In the African population, the prevalence of fecaliths in patients whose appendices were palpated incidentally was four per cent versus 23% for those with appendicitis (p = 0.04). The difference in prevalence of incidental appendiceal fecaliths in the two populations was statistically significant (p less than 0.005). The prevalence of fecaliths is higher in developed countries, such as Canada, than in developing countries, such as Africa, and is also higher in patients with than in those without appendicitis. These data support the theory that the low-fiber diets consumed in developed countries lead to fecalith formation, which then predisposes to appendicitis. PMID- 2990362 TI - Glomus tumors of the hand. Presentation of fourteen cases. AB - The authors report a series of fourteen patients with a glomus tumor in the hand. The major symptoms were excruciating spontaneous pain upon mechanical and thermal stimulation of the tumor. In subungual tumors, a blue discoloration is suggestive of diagnosis; but this was found in only 50 percent of cases. The difficult in correct diagnosis and therefore for complete removal is due to in the location of the tumor. Recurrence may occur in cases of incompletely removed tumor. In three cases, the tumor was submitted to electron microscopic examination which led to a better comprehension of the histogenesis of these tumors. PMID- 2990361 TI - Bile duct carcinoma. Diagnosis and treatment. AB - Fifty-three patients were evaluated for carcinoma of the extrahepatic bile ducts at the University of Virginia from 1951 to April 1984. This population was retrospectively reviewed and 33 preoperative and postoperative variables were analyzed to evaluate the predictors of increased survival. No preoperative data, including symptoms, admitting laboratory data, or tumor location, predicted increased survival. Whipple resection yielded a median survival of 12 months; palliative resections, 1.5 months; and laparotomy, only 5.5 months; these differences were not statistically significant. Experience with six patients treated with internal radiation (iridium-192), plus as much as 4000 rads of external beam irradiation, suggests that this combination of radiation treatment may aid palliation. PMID- 2990363 TI - Subungual glomus tumors. Value of the direct dorsal approach. AB - The authors operated upon 4 subungual tumors between november 1975 and july 1976. In three cases, the tumor was located under the center of the nail and a direct dorsal approach--atraumatic detachment of the nail and approach via a longitudinal incision of the nail bed--was employed. Excision was performed using microsurgical technique. After trimming and perforation (classical techniques), the nail was replaced and fixed. This approach clearly identifies the boundaries of the tumor and facilitates radical removal. The cosmetic result is excellent. No recurrences have been seen with a 5 year follow-up. PMID- 2990364 TI - Suloctidil treatment prevents the development of post-traumatic feline spinal cord ischemia. AB - The effects of suloctidil (1.0 mg/kg i.v.) on white matter spinal cord blood flow (SCBF) and somatosensory evoked potential (SEP) conduction were assessed in a feline lumbar spinal cord contusion model. SEP conduction ceased immediately after a 500 g-cm injury and did not recover. SCBF following suloctidil declined by only 12% from pre-injury levels over the 4 hr experiment as compared to a 42% decline observed following vehicle injection. Accordingly, 7/8 vehicle cats, but only 1/5 suloctidil-treated cats, had 4 hr SCBF values of less than 10 ml/100 g/min. Bradycardiac and hypotensive effects of suloctidil were also noted. The ability of suloctidil to prevent post-traumatic ischemia may be useful in the acute treatment of spinal cord injury. PMID- 2990365 TI - Thromboxane A2-induced vascular endothelial cell damage and respiratory smooth muscle cell contraction: inhibition by flunarizine, a Ca2+-overload blocker. AB - The fast intravenous injection of arachidonic acid (AA) in mice produces, in a dose-related way, mortality due to respiratory distress. Upon electron microscopical examination an extensive oedematous damage of the capillary endothelium was found; thrombotic platelet obstructions were present in a minority of pulmonary capillaries only. Protection against this toxic AA-effect is obtained with inhibitors of fatty acid cyclo-oxygenase and of thromboxane (TXA2) synthetase, suggesting involvement of TXA2 as a causative mediator. The Ca2+-entry blockers flunarizine, niludipine and nimodipine, not affecting TXA2 synthesis by murine platelets, also provide protection, but not the antiplatelet drugs ticlopidine, dipyridamole or suloctidil; thrombocytopenia induced by busulphan does not affect the AA-induced mortality nor the protection obtained with flunarizine. Platelet-independent bronchoconstriction induced by AA in guinea-pigs is also inhibited by flunarizine. This study suggests that the AA induced mortality test reflects pulmonary conversion of AA to TXA2 producing endothelial cell damage and respiratory smooth muscle cell contraction rather than a thrombotic phenomenon. The protective effect of flunarizine against TXA2 induced changes in vivo may contribute to its effectiveness in particular hypoxic conditions associated with liberation of AA. PMID- 2990366 TI - Beta-adrenergic receptors in rat myocardium during the development and reversal of hypertrophy and following chronic infusions of angiotensin II and epinephrine. AB - The effect of cardiac hypertrophy on beta-adrenergic receptor density and affinity was studied under 4 experimental conditions: in the spontaneously hypertensive rat (SHR), in the 2K-1C renal hypertensive rat (RHR), and following subcutaneous infusions of 2 pressor agents; epinephrine (E) and angiotensin II(AII). Using the antagonist 3H-dihydroalprenolol [( 3H]-DHA), the number of binding sites was shown to significantly decrease at both 13 and 24 weeks of age in the SHR when compared to age-matched WKY, with no change in affinity. In the RHR a significant increase in binding sites was observed at both 6 and 10 weeks after clipping, returning towards normal levels following removal of the clipped kidney. Cardiac hypertrophy and hypertension were induced by subcutaneous infusions for up to 2 weeks of both E and AII. E caused an alteration in receptor density, causing a significant decrease with no change in affinity. In contradistinction, although the degree of hypertrophy was the same following AII, no changes in receptor density or affinity were seen. These present experiments confirm our hypothesis that different models of hypertensive hypertrophy are associated with varying changes in beta-adrenergic receptors. This suggests that any consequential changes in myocardial function may be a result of other post receptor mechanisms. PMID- 2990368 TI - ATPase and cytochrome oxidase activities at the polar organelle in swarm cells of Sphaerotilus natans: an ultrastructural study. AB - The polar organelle of bacteria presumably is part of the flagellar apparatus. In order to characterize this structure, cytochemical studies on Sphaerotilus natans have been performed. Marked ATPase activity is associated with the inner boundary layer and central layer of this organelle. The spaces between the boundary layers and the central layer of the polar organelle which are traversed by fine fibrilles are positive for reactions with diaminobenzidine. This indicates cytochrome oxidase activity. S. natans possesses a ribbon-like, helically shaped polar organelle which is divided concomitantly with cell fission, possibly explaining inheritance of this structure and of the flagellar apparatus. PMID- 2990369 TI - More on birth seasonality and schizophrenia. PMID- 2990367 TI - The major soluble cytochromes of the obligately aerobic sulfur bacterium, Thiobacillus neapolitanus. AB - Four cytochromes were isolated from soluble extracts of the aerobic sulfur bacterium, Thiobacillus neapolitanus. The two most abundant proteins were purified to homogeneity and thoroughly characterized. Cytochrome c-554 (547) is a monomeric, small molecular weight protein which is unusual in having two well resolved alpha peaks in UV-visible absorption spectra. The redox potential is 208 mV. Native cytochrome c-549 is oligomeric, but has a subunit size of about 26,000. The yield of this protein could be improved dramatically by washing membranes with 30% ammonium sulfate, but the material solubilized by this method had a larger native molecular weight than that in the initial 0.1 M Tris-Cl extract and behaved differently on chromatography. The properties of cytochrome c 549 including subunit size and UV-visible absorption spectra are similar to mitochondrial cytochrome c1 and chloroplast cytochrome f, which suggests that it may be a modified form of the predominant membrane cytochrome. Based on cytochrome content, it is suggested that T. neapolitanus is not closely related to other thiobacilli. PMID- 2990370 TI - Bile duct adenoma. PMID- 2990371 TI - Perineurioma. PMID- 2990372 TI - Small-cell undifferentiated carcinoma of the rectosigmoid colon. AB - We describe three patients with small-cell undifferentiated carcinoma of the rectosigmoid colon. Though there was a wide range of patient age, history, and gross presentation of the malignancy, there was overlap in the histopathologic appearance and the biological behavior of the tumor. All three cases had liver involvement at the time of diagnosis. Despite multidrug chemotherapy, two patients died within a year of diagnosis, and the third patient had progressive disease. Ultrastructurally, all cases had membrane-bound dense-core granules, however, there was considerable variation of size and number of granules among the tumors, as well as other forms of endodermal differentiation. Whether small cell undifferentiated carcinoma presents as a small focus within a neoplastic polyp or as a large primary tumor, the therapy must include a systemic aggressive protocol. PMID- 2990373 TI - Congenital giant axonal neuropathy. AB - Giant axonal neuropathy (GAN) is a distal sensorimotor neuropathy, characterized by neurofilamentous axonal swellings, with usual onset at 2 to 3 years of age. We report a case of congenital GAN with hypotonia at birth. At 7 months of age, nerve conduction studies showed almost complete lack of sensory and motor responses in the lower extremities. A sural nerve biopsy specimen disclosed absence of myelinated axons. Autopsy, following death at 15 months of age, revealed axonal swellings in peripheral nerves and distal degeneration of long spinal cord tracts. The neurofilamentous content of the axonal swellings was confirmed by Glees-Marsland staining and immunoperoxidase reaction with antibodies to neurofilaments. Axonal swellings did not stain with periodic acid Schiff and were not seen in the cerebral cortex or brain stem, distinguishing this process from infantile neuroaxonal dystrophy. This patient illustrates congenital GAN with subsequent rapid progression. PMID- 2990374 TI - An 18-year history of a corticotropin-secreting spindle cell carcinoid in the lung. AB - A unique case is reported demonstrating latency and slow growth of a spindle-cell carcinoid in the lung. Seventeen years after bilateral adrenalectomy, performed for Cushing's syndrome, a patient presented with a newly developed lung tumor. The tumor demonstrated spindle cell histology, neurosecretory granules typical for carcinoid, and a positive immunoreactivity for corticotropin. After the removal of the lung tumor, the high serum corticotropin level returned to normal. PMID- 2990375 TI - Embryonal (undifferentiated) sarcoma of the liver. Fine-needle aspiration cytology and ultrastructural findings. AB - We present a case of embryonal (undifferentiated) sarcoma with light microscopic and electron microscopic findings. The ultrastructural appearances are similar to the few isolated cases reported previously and include features suggesting myofibroblast involvement. Fine-needle aspiration biopsy proved to be of value in confirmation of persistent tumor. PMID- 2990376 TI - Primary adenoid cystic carcinoma of the skin. PMID- 2990377 TI - Clinical comparison of glucose quantitation methods. AB - Quantitation of glucose in human serum, plasma, or whole blood by several commercially available methods and a manual hexokinase method were compared in various combinations. The techniques were selected as being representative of those available in large and small laboratory settings. Each differs as to reagent configuration and/or nature of sample required, such that a particular technique is more or less desirable depending on the unique situation of the user. Differences in relative performances in respect to accuracy and precision were revealed. Some are statistically significant and medically important. These differences must be appreciated by professionals of both large and small clinical laboratories, the managing physicians, and the patients. PMID- 2990378 TI - Acquired immunodeficiency syndrome. Clinicopathologic study of 56 autopsies. AB - We studied 56 patients with acquired immunodeficiency syndrome who underwent autopsy at the New York University Medical Center. We uncovered several entities that were not previously described or emphasized in prior series. These were as follows: (1) three cases of necrotizing arteritis with cytomegalovirus inclusions; (2) dissemination in 95% of cases of Kaposi's sarcoma; (3) unusual clinical and histologic presentations of Kaposi's sarcoma presenting diagnostic difficulties for both clinicians and pathologists; and (4) a very high incidence of serious nonmycobacterial infections. In addition, previous autopsy studies have disagreed over such matters as the incidence of disseminated candidiasis, hemophagocytosis, and severe adrenal necrosis in acquired immunodeficiency syndrome. All of these studies, except one, involved ten to 12 cases each. Our study of 56 cases helps to resolve these disagreements. PMID- 2990379 TI - Immediate causes of death in acquired immunodeficiency syndrome. AB - We evaluated the immediate causes of death in 54 adults who underwent an autopsy and were diagnosed as having died of the acquired immunodeficiency syndrome between April 1980 and October 1983. The study group included 25 Haitians, 19 homosexual men, five intravenous drug abusers, two hemophiliacs (type A), and three with no known risk. Fourteen died of central nervous system diseases: 11 of Toxoplasma encephalitis, one of progressive multifocal leukoencephalopathy, one of viral encephalitis, and one of intracerebral hemorrhage. Thirty died of respiratory failure; 16 of Pneumocystis carinii pneumonia, ten of cytomegalovirus pneumonia, one of multiple infections, one of interstitial pneumonia, and two of bacterial pneumonia. Two died of overwhelming generalized infections: one of Mycobacterium avium-intracellulare and one of listeriosis. Six died of disseminated Kaposi's sarcoma, while the remaining two persons died of Toxoplasma myocarditis (one) and one of shock resulting from a percutaneous liver biopsy, respectively. There were differences in the immediate causes of death between Haitians and homosexuals as follows: 63% of homosexual men died of either P carinii pneumonia or Kaposi's sarcoma vs 20% of Haitians. In contrast, 72% of Haitians died of other opportunistic infections as compared with 21% of homosexuals. There has not been an increase in the proportion of cases diagnosed premortem since 1982 and overall, only 32 (58%) were diagnosed premortem; the rest were diagnosed only at autopsy. This study provided evidence that 42% died of currently untreatable diseases. PMID- 2990380 TI - Synthesis of leukotriene B3. PMID- 2990381 TI - Neuromuscular problems in the burn patient: cause and prevention. AB - Neuromuscular complications of the burn patient that occur during hospitalization frequently are overlooked. Eighty-eight patients with signs of weakness or complaints of easy fatigability were examined clinically and electrodiagnostically. The most frequently diagnosed neuromuscular abnormality in this study was generalized peripheral neuropathy. Other specific neuromuscular problems, in order of frequency of occurrence, were found to involve the deltoid muscle, peroneal nerve, ulnar nerve, median nerve, and brachial plexus. These lesions are found to be commonly due to (1) poor positioning, both in bed and in the operating room, and (2) heavy bulky dressings over superficial nerves. Thus, it is concluded that many of these injuries offer a significant potential for prevention through attention to physiologic positioning and meticulous patient care. PMID- 2990382 TI - Systemic dissemination of ethanolamine oleate after injection sclerotherapy for esophageal varices. AB - Sclerotherapy for esophageal varices was performed on six patients for a total of 16 injections. The sclerosant contained 5% ethanolamine oleate and technetium Tc 99m sodium pertechnetate solution at the volume ratio of 9:1. At each injection session 5 to 20 mL of this solution was injected into one varix. The distribution of the injected material was observed by a scintillation camera. Systemic dissemination of the sclerosant through the portal vein was demonstrated in five procedures, in which the injections were correctly intravenous. In three dogs 4 mL of 5% ethanolamine oleate made from oleic acids labeled with iodine 131 was injected into the gastric coronary vein. Most of the substance injected was washed out within five minutes, and systemic dissemination became evident. PMID- 2990383 TI - Why do patients with postsurgical acute tubular necrosis die? AB - We compared early (less than 70 days) cumulative survival, cause of death, and morbidity (defined as the number of organ system failures and reoperations) of 236 patients who needed dialysis for acute tubular necrosis (ATN) after transplantation with that of a control group of 215 transplant patients who had immediate renal function after transplantation. The patients were matched for age, number of transplants, time of transplantation, diabetic status, living related vs cadaveric donor, and number of HLA matches. There were no differences in cumulative survival after transplantation, causes of death, organ system failure, or surgical complications as evident from the number of reoperations. The ATN patients' overall clinical course was similar to that described for other postsurgical patients with ATN in their native kidneys. We conclude that the poor survival and the surgical complications in patients with postoperative ATN are due to the basic disease and not to acute renal failure or to dialysis. To improve survival, research should be focused on the basic disease and the surgical procedures rather than on dialysis technique. PMID- 2990384 TI - Meso-2,3-dimercaptosuccinic acid increases the inhibition of glutathione S transferase activity from rat liver cytosol supernatants by di-n-butyltin dichloride. AB - Di-n-butyltin dichloride (DBTC) and tricyclohexyltin chloride (TCHTC) inhibited the glutathione S-transferase activity of rat liver cytosolic supernatants towards 1-chloro-2,4-dinitrobenzene. Meso-2,3-dimercaptosuccinic acid increased inhibition of glutathione S-transferase by DBTC but lowered inhibition of the enzyme by TCHTC. PMID- 2990385 TI - Analysis of the HSV-2 early AG-4 antigen. AB - Genital herpes simplex virus type 2 (HSV-2) infections can be distinguished from present or past HSV-1 infections by an AG-4 antigen complement fixation assay. The assay which utilizes a 4 hour HSV-2 infected cell extract prepared at a multiplicity of infection (MOI) of 1.0 PFU/cell, appears to consist of several viral proteins. Studies using monoclonal antibodies, polyclonal rabbit hyperimmune serum, HSV-1 X HSV-2 intertypic recombinant viruses and polyacrylamide gel electrophoresis suggest that ICP 8 may be one of the major antigens involved in the complement fixing reaction. It is probable that the success of the assay is not due to a true type specificity but rather a threshold phenomenon in which HSV-2 extracts contain more early viral antigens (including ICP 8) and sera from HSV-2 patients contain more complement fixing antibody to these antigens. PMID- 2990386 TI - Molecular cloning and partial characterization of the coxsackievirus B3 genome. Brief report. AB - The RNA genome of coxsackievirus B3 has been cloned and partially characterized by restriction mapping, partial sequence analysis, and hybridization to heterologous coxsackievirus B genomes. It differs significantly from the poliovirus genomic structure. PMID- 2990387 TI - Two modes of human rotavirus entry into MA 104 cells. AB - Entry of the KUN strain of human rotavirus into MA 104 cells was studied by electron microscopy. Virus particles attached to the cell membrane appeared to be almost exclusively double-shelled virions. These attached virions followed two distinct pathways into the cytoplasm depending on pretreatment with trypsin. Using infectious rotavirus which had been pretreated with trypsin, the viral nucleoids passed directly into the cytoplasm within 5 minutes after inoculation, through dissolution of the viral capsid and cell membrane. Using non-infectious rotavirus that had not been pretreated with trypsin, phagocytosis or pinocytosis occurred in which virions were sequestered into lysosomes 20 minutes after virus attachment to the cell membrane. After being sequestered, uncoating of the rotavirus virions within lysosomes was seen, but it did not result in release of the genome. On the basis of these observations it was concluded that when virions were pretreated with trypsin, virus replication occurred following the direct passage of viral nucleoids into the cell cytoplasm. However, mere phagocytosis of virus particles into cell lysosomes, which occurred when trypsin-untreated virus was used, does not appear to be related to rotavirus replication. PMID- 2990388 TI - The proteins expressed in vivo and in vitro by an orbivirus of the Kemerovo serogroup isolated from Ixodes uriae ticks from St. Abb's Head, Scotland. AB - The genome of FT363 virus, a member of the Kemerovo serogroup of orbiviruses, was shown to consist of 10 segments of dsRNA. Examination of the intracellular proteins expressed by this virus in Vero cells showed the presence of 9 major and several minor products. Similar results were obtained by translation of the virus dsRNA in a rabbit reticulocyte lysate. Comparison of in vivo and in vitro products by immunoprecipitation using polyclonal FT363 ascitic fluid showed a difference in the numbers of polypeptides precipitated. The significance of these results is discussed. PMID- 2990389 TI - European isolates of bovine herpesvirus 1: a comparison of restriction endonuclease sites, polypeptides, and reactivity with monoclonal antibodies. AB - Eleven European isolates of bovine herpesvirus 1 (BHV-1), together with two reference virus strains were compared by restriction endonuclease digestion, by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and by their reactivity with a panel of monoclonal antibodies (McAb's). Based on the cleavage pattern of viral DNA with the restriction endonuclease Hind III the strains could be assigned to one of two established major virus types. Analysis by SDS-PAGE of viral polypeptides revealed that four protein species either displayed virus type or subtype specific minor variation of migration characteristics. Of 43 McAb's tested all reacted with all type 1 strains, whereas five antibodies failed to recognize some of the type 2 viruses. The existence of type specific variations among virus specified proteins was further evidenced by the recovery of one McAb recognizing type 1 viruses only. The data show that BHV-1 isolates can be assigned to established virus types according to the SDS-PAGE profile of viral proteins or the selective reactivity with type specific McAb's. PMID- 2990390 TI - Studies on the relationship between coronaviruses from the intestinal and respiratory tracts of calves. AB - An immunofluorescence test on smears of nasal epithelial cells was used to detect coronavirus infection in the respiratory tract of calves. Thirteen gnotobiotic calves were infected with coronavirus isolates derived from faeces or respiratory material: virus was detected in faeces and nasal swabs from all animals. In 115 calves from a field survey, there was a significant association between coronavirus excretion from both respiratory and enteric routes in calves with diarrhoea. In a further 12 calves, at necropsy, the predilection sites for coronavirus growth were the distal small intestine, large intestine and the epithelia of the nasal cavity and trachea. Antigen was not found in lung tissue by immunofluorescence or immunoperoxidase staining. Infection with enteric coronavirus induced immunity to reinfection and to heterologous challenge with two coronavirus isolates derived from the respiratory tract. Nine coronaviruses were cultivated, cloned and antisera to three were prepared in pigs. There was complete virus neutralisation in tests with homologous sera and significant cross reactions with the eight other isolates which were of intestinal and respiratory origin. Thus, these bovine coronavirus isolates belonged to the same serotype despite the source of virus. PMID- 2990391 TI - Effects of activated serum factor on the induction of Epstein-Barr virus antigens and cell differentiation. AB - n-Butyrate and 12-0-tetradecanoylphorbol-13-acetate (TPA) have been shown to be potent inducers of Epstein-Barr virus (EBV) antigen synthesis (10, 18) and cell differentiation (1,2). Activated serum factor (SF) has also been reported to induce virus antigen synthesis and moreover it is capable of cooperative action with other inducers (5). We have demonstrated with an EBV-carrying lymphoma cell line (Raji) that activated SF can also induce the appearance of some cellular alterations, specific for early steps of the productive virus cycle in a low proportion of cells. In addition cell differentiation towards plasma cell was noted in about 8 per cent of SF-treated cells. It was confirmed that activated SF enhances the induction of virus antigens by n-butyrate or TPA as detected by the immunofluorescence technique. It was shown that it also potentiates the development of virus-induced changes in cellular morphology and the cell differentiation induced by these drugs. This effect was more pronounced in combination with TPA than with butyrate. Possible relationships between cell differentiation and virus-antigen expression are discussed. PMID- 2990393 TI - [Cerebellar glioblastoma multiforme: report of a case in a child]. AB - The authors relate a case of glioblastoma multiforme of the cerebellum in a nine years old girl. They correlate this case with reports published and they think that this pathology is not so rare as believed. This is the second case report, in Brazil, in children. PMID- 2990394 TI - Infectious origin and Epstein-Barr virus. PMID- 2990392 TI - Crossed immunoelectrophoretic analysis of herpes simplex virus type 2 proteins. Characterization of antigen-5. AB - Herpes simplex virus type 2 proteins extracted from infected cells and analysed by crossed immunoelectrophoresis identified a nonglycosylated antigen named Ag-5. The antigen contained two proteins when extracted from the agarose gel and the molecular weights were 128K and 91K. Both proteins are located in the nucleus of the infected cells and the 128K is identical to ICP-8. The 91K protein is based on the reactivity with monoclonal antibodies most likely the alkaline exonuclease mapped by Preston and Cordingly (25). Our data show that although the proteins ICP-8 and 91K coprecipitate they differ in both peptide composition and in immunological specificity. PMID- 2990395 TI - Early prenatal diagnosis of inborn error of metabolism: a case report of a fetus affected with Fabry's disease. PMID- 2990396 TI - Viral infections of the respiratory tract. PMID- 2990397 TI - AIDS screening test now available to everyone. PMID- 2990398 TI - Experimental infection with Murray Valley encephalitis virus. Pigs, cattle, sheep, dogs, rabbits, macropods and chickens. AB - A total of 142 young animals including 10 domestic and 14 feral pigs, 12 Hereford calves, 12 crossbred and 24 Merino lambs, 11 dogs, 8 domestic and 16 feral rabbits, 14 Grey kangaroos, 9 Agile wallabies and 12 chickens was exposed to infection with 4 strains of Murray Valley encephalitis virus (MVE), mainly using orally infected Culex annulirostris mosquitoes. In terms of their viraemic response, the animals were grouped into high (Grey kangaroos, rabbits), moderate (pigs, dogs, chickens) and low (calves, lambs, Agile wallabies). Recipient Cx annulirostris induced to bite these animals 1-7 d post-inoculation exhibited infection rates up to 95% but usually were much lower. Some animals in which virus was not detected by intracerebral inoculation of suckling mice were capable of infecting up to 5% Cx annulirostris. Haemagglutination-inhibiting antibody (HI) was of high titre and persistent in rabbits and Grey kangaroos but moderated in others, e.g., cattle, sheep, Agile wallabies to be transient and of low titre. Cattle and Merino lambs were often completely non-responsive. This study increases the range of vertebrates that could be considered as maintenance or amplifying hosts of MVE and, on the basis of their HI antibody responses, evaluates them as potential sentinels of infection in Australia. PMID- 2990399 TI - Aetiology and pathogenesis of mucosal disease of cattle: current concepts, observations and speculation. PMID- 2990400 TI - Discharge characteristics of motor units and the surface EMG during fatiguing isometric contractions at submaximal tensions. AB - The RMS amplitude of the surface electromyogram (EMG) and the frequency of discharge of motor units was examined throughout the duration of isometric contractions of the adductor pollicis muscles sustained to fatigue at tensions of 25, 40, and 55% of the maximum voluntary strength (MVC) of eight male subjects during fatiguing isometric contractions. The maximum strength of the muscle and the EMG above the adductor pollicis muscles was also assessed during 3 s of voluntary and electrically induced isometric contractions interposed at 25, 50, 75, and 100% of the duration of the fatiguing contractions. At the point of fatigue from submaximal isometric contractions, the RMS amplitude of the surface EMG was highest for contractions at 55 as compared to 40 and 25% MVC. The lower RMS amplitude of the EMG during contractions at lower as compared to higher tensions at the point of fatigue was paralleled by a lower discharge frequency of the alpha motor neurons in the fatigued muscle during contractions at 25% as compared to 40 and 70% MVC. The reduction in discharge frequency was probably of a sufficient order of magnitude to account for the lower amplitude of the EMG at the end of fatiguing isometric contractions at lower tensions. PMID- 2990401 TI - The concept, physiopathology. Its role in understanding functional disorders. PMID- 2990402 TI - The nature of stress. PMID- 2990403 TI - Stress and nursing practice. PMID- 2990404 TI - Role of the pi initiation protein and direct nucleotide sequence repeats in the regulation of plasmid R6K replication. PMID- 2990405 TI - Initiation of replication of the Escherichia coli chromosomal origin reconstituted with purified enzymes. AB - A mixture of purified proteins has replaced a crude enzyme fraction capable of efficient replication of oriC-containing plasmids. The reconstituted enzyme system contains proteins which provide initiation, replication, and specificity functions required for dnaA-dependent replication specific for an oriC template. Replication can be separated into successive stages of RNA synthesis and DNA replication. Isolation of an intermediate no longer requiring RNA polymerase action requires the presence of dnaA protein, DNA gyrase, dnaB protein and dnaC protein. Intermediate formation likely involves binding of dnaA protein to a 9-bp sequence present 4 times as inverted repeats within the chromosomal origin sequence. PMID- 2990407 TI - Report on a workshop: structure and function. PMID- 2990406 TI - Origin and initiation sites of lambda dv DNA replication in vitro. AB - The sequence of lambda DNA essential for the unique initiation of replication was analyzed in an in vitro replication system. Fragments of lambda DNA were inserted into pBR322 and used as templates or were circularized in vitro in the absence of pBR322 and employed in the same tests. A 165-bp region left of the EcoRI site in the O gene of the lambda genome was defined as the functional origin. This region, which we defined as the ori region, carries, in order, the 4 19-bp repeat sequences where the O protein binds (ori-repeats), an A+T-rich stretch, and a region that constitutes part of a large palindromic structure. Regions that have long been suspected to participate in lambda DNA replication initiation, ice and oop were not required for the O, P-dependent lambda-specific replication initiation. The lambda dv and the "ori region plus" recombinant plasmids initiated DNA synthesis at or around this region, and the reaction depended on the presence of lambda-coded initiators, O and P proteins. Early replicative intermediates of lambda dv were prepared in an in vitro replication system in the presence of ddCTP, an inhibitor of DNA chain elongation. This system allows only that DNA synthesis that is a result of replication initiation. Using this system, the initiation site(s) of the DNA synthesis was finely analyzed by mapping the transition sites from primer RNA to DNA synthesis. Short-chain DNAs produced from regions near the ori region were purified from the intermediates. A fraction of the short-chain DNAs was covalently linked to primer RNA. The 5'-ends that had been linked to RNA (transition sites) were exposed by alkaline hydrolysis, labeled with 32P, and the transition sites were mapped along the nucleotide sequence of the genome. Two striking features emerged from this analysis: (i) The transition sites are located on both sides of the ori region, and no transition arose within the 165-bp ori region; (ii) The transition sites on both sides are not unique, but multiple, and are clustered in one of the 2 strands. Furthermore, their orientation demonstrates that the DNA synthesis in initiation of replication from the 2 sides of the ori region converge. The frequency of the "leftward" DNA synthesis is several times higher than that of "rightward" synthesis. These results reflect asymmetric bidirectional replication of lambda dv DNA, and may also reflect replication of lambda phage DNA. PMID- 2990408 TI - Broad host-range plasmid R1162: replication, incompatibility, and copy-number control. PMID- 2990409 TI - Evolutionary relevance of genetic rearrangements involving plasmids. PMID- 2990410 TI - The partition functions of P1, P7, and F miniplasmids. AB - The partition regions of P1, P7, and F miniplasmids are discrete DNA sequences of about 3 kb in length that will promote accurate partition of hybrid plasmids independent of the source of replication functions or the position or orientation of the elements. Each of the par regions seems to be very similarly organized, with open reading frames for essential proteins and a terminal site which appears to be analogous to the centromere of eukaryotic cells. When cloned, these terminal sites exert incompatibility against their respective parent plasmids presumably because they can compete with the parent plasmids as substrates for partition. We have determined the complete DNA sequence of the P1 par region. In addition to the open reading frame for the essential parA protein (42-44 kd), the region contains a second open reading frame which could encode a 38-kd protein. The 2 large open reading frames appear to form an operon that is negatively regulated from a site adjacent to the promoter and responds to the par gene products in trans. Both this site and the downstream "centromere" site, incB, contain blocks of extremely AT-rich sequences, which are postulated to be binding sites for par proteins. The incB and upstream AT-rich regions both contain 20-bp imperfect inverted repeats. Further downstream from the minimal incB sequence (172 bp) lies an additional region which is essential for partition. The further analysis of the P1 par region should be greatly facilitated by the finding that it can function in cis to stabilize pBR322 vectors under conditions where the copy number of pBR322 is reduced. PMID- 2990411 TI - Genetic interactions of broad host-range plasmid RK2: evidence for a complex replication regulon. AB - The kil and kor genes of RK2 are novel genetic determinants further that the kil and kor network constitutes a replication regulon, and that perhaps the function of this regulon is to ensure expression of trfA at appropriate levels. The complexity of this regulon may reflect an ability of the system to adapt to the intracellular environments of a variety of hosts. Indeed, there is tantalizing evidence that regions encoding kil or kor genes are important to host range (1,2,6,28; Schmidhauser and Helinski, pers. comm.). We are therefore hopeful that the study of these genes and the eventual determination of the molecular basis of their actions will lead to a complete understanding of the replication control and broad host range capability of IncP plasmids. PMID- 2990412 TI - Replication determinants of the broad host-range plasmid RSF1010. PMID- 2990413 TI - Regulation of replication and maintenance functions of broad host-range plasmid RK2. AB - Replication of broad host-range plasmid RK2 depends on a cisacting vegatative replication origin oriVRK2 and the polypeptide product(s) of the trans-acting gene trfA as well as on host-specified products. The trfA gene is the second cistron in a polycistronic unit whose first cistron may be kilD, one of 4 known RK2-specified kil loci (kilA, B, C, and D) which are inhibitory for bacterial host or plasmid vector in the absence of kor functions which suppress in trans the effect of their respective kil genes. Transcription of the operon containing trfA is negatively regulated by the products of both the trfB locus (alias korD and korA) and korB. The loci, trfB and korB, are expressed from a single transcriptional unit which we propose to be negatively autoregulated by the products of both loci, although an additional, weaker and unregulated transcript may also express korB. While deletions in the oriVRK2 region have indicated the presence of copy number control elements adjacent to and possibly overlapping with the minimal oriVRK2 segment, the overriding control of copy number seems to reside in the trfB and korB loci which in conjunction appear to reduce expression of the trfA gene to levels limiting for replication. Coregulation of trfA with kil genes may indicate that kil genes play a role in plasmid maintenance other than replication. PMID- 2990414 TI - Replication control for pT181, an indirectly regulated plasmid. AB - PT181 is a fully sequenced Staphylococcus aureus plasmid whose size is 4,437 bp. It specifies tetracycline resistance and has a copy number of about 22 per cell in exponentially growing cultures. The functional organization of the pT181 replicon is centered around the coding sequence for a 35-kd protein, RepC, that is absolutely required for replication of the plasmid. The replication origin is contained within the repC coding sequence and the region immediately 5' to the RepC start is involved in control of the plasmid replication rate. PT181 replication is controlled at the level of RepC synthesis by a negative regulatory system that is functionally similar to that of the Co1E1 and IncFII plasmids of Escherichia coli. The pT181 control circuit involves 2 short transcripts, RNA I and RNA II, that are transcribed from the region specifying the 5' end of the untranslated repC mRNA leader and in the opposite direction. These are referred to as countertranscripts. The countertranscripts regulate RepC synthesis by a mechanism that probably involves interaction with the repC mRNA leader in a manner that interferes with translation. Both of the countertranscripts seem to be necessary for normal replication control; their separate roles remain unclear. Unlike plasmids of the Co1E1 and IncFII groups, plasmids such as Co1E1 are considered to have direct regulation of replication because the inhibitory element of the copy control circuit directly inhibits the initiation of replication. Plasmids such as pT181 are considered to have indirect regulation of replication because the product of the regulated step, RepC, is trans-active. Plasmids of the IncFII type are considered to have direct regulation of replication because the product of the regulated step, RepA is cis-active The analysis of pT181 replication physiology has illustrated 2 important differences between directly and indirectly regulated plasmids: a) for directly regulated plasmids, copy mutants specifying a normal inhibitor substance but an inactive target site exclude the wild-type or recessive mutants by directly interfering with their replication. Analogous mutants of indirectly regulated plasmids coexist readily with the wild-type and all mutants (although they do manifest segregational incompatibility) because the Rep protein is always shared by all plasmids in the cell, regardless of its source. b) Mutations of directly regulated plasmids in the region where target transcript and countertranscript overlap may give rise to totally new incompatibility groups because they engender independently self-correcting copy pools.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990415 TI - Mechanisms of transposition in bacteria. AB - The transposable elements of bacteria are diverse in size, functional arrangement, DNA sequence, and in their modes of transposition. We review here data suggesting that the kanamycin-resistance transposon Tn5 moves without replicating (conservative transposition), but the ampicillin-resistance transposon Tn3 is duplicated when it transposes, and that both the chloramphenicol-resistance transposon Tn9 and bacteriophage Mu are replicated in some events but not in others. A model is presented in which conservative and replicative transposition are alternative branches of a single pathway. PMID- 2990416 TI - Incompatibility and IncFII plasmid replication control. AB - The DNA coding for replication control and incompatibility of the plasmid NR1 serves as a template in vivo and in vitro for RNA transcription in both directions. In the rightward direction, RNA synthesis begins from 2 different promoters, one of which is regulated and the other constitutive. In vivo, each of these transcripts is more than 1,000 nucleotides long, terminating near the estimated site for the origin of replication. These transcripts serve as messenger RNA for several proteins. One protein (repA1) is required for replication and another (repA2) serves as the repressor for the regulated rightward promoter. RNA synthesis in the leftward direction is constitutive and produces a single transcript of 91 nucleotides which is complementary in sequence to the rightward transcripts. This small transcript is the incompatibility product which regulates the replication of the plasmid. When the intracellular concentration of the small transcript is experimentally varied, the rate of translation of the rightward transcripts and the rate of initiation of replication (plasmid copy number) vary inversely to its concentration. The mode of action of this inhibitor RNA is likely to be formation of an RNA-RNA duplex with the rightward transcripts, thereby inhibiting the translation which would produce the required replication protein. The probability that a rightward transcript will escape interaction with the small RNA molecules and thus allow replication to initiate can be predicted from the laws of mass action based on base-stacking free energies for the likely sequences of initial contact. The estimated intracellular RNA concentrations, based on quantitative hybridization experiments, are agreement with those predicted from the calculated equilibrium constants for duplex formation. PMID- 2990417 TI - P1 plasmid maintenance: a paradigm of precise control. PMID- 2990418 TI - Partitioning of the pSC101 plasmid during cell division. PMID- 2990420 TI - Positive regulation and transcription initiation of xyl operons on TOL plasmid. PMID- 2990419 TI - DNA-protein interaction at the replication origins of plasmid chromosomes. AB - Novel techniques have been developed to purify replication initiator proteins of the plasmids R6K and pSC101. The techniques consist of tagging the initiator cistrons at the C-terminus with beta-galactosidase-encoding DNA of Escherichia coli in the correct translational phase. The hybrid proteins are then rapidly purified by adsorption to and elution from a beta-galactosidase- specific affinity column. Two procedures have been devised to isolate the nonfused initiator proteins using the fused protein as a handle. The first procedure, called subunit association chromatography, exploits the association of a monomer of nontagged protein with that of beta-galactosidase-tagged protein in isolating both types of proteins by beta-galactosidase specific affinity column chromatography. The second procedure involves the fusion of the initiator protein to beta-galactosidase via a specific linker DNA. The linker DNA encodes a protein which is readily and specifically hydrolyzed by a sequence specific protease, thus releasing the initiator protein from beta-galactosidase. Using purified or partially purified initiator protein, we have demonstrated that the R6K encoded initiator protein (Pi protein) binds to a consensus 22 bp sequence at 2 regions of the plasmid chromosome. The pSC101-encoded initiator protein binds to sequences at or near the plasmid replication origin. At low concentrations the protein binds to a nucleation site and upon raising the concentrations of the protein binding is promoted at 4 adjacent sequences that have partial homologies with the nucleation sequence. Deletion of the binding site leads to a nonfunctional replication origin. PMID- 2990421 TI - Linear plasmids with terminal inverted repeats obtained from Streptomyces rochei and Kluyveromyces lactis. AB - Linear plasmids with inverted terminal repeats of 614 bp were obtained from Streptomyces rochei which produced lankacidin. The 5' ends were blocked by the association of a terminal protein. A DNA model of racket frame-like structure is presented implying the juxtaposition of 2 double-stranded DNAs of the same sequence through the binding of cohesive proteins which recognize and bind to the DNA. Two linear plasmids with the inverted terminal repeats of 202 and 184 bp were obtained from a yeast, Kluyveromyces lactis. A killer toxin was produced from the shorter plasmid. The protein toxin inhibited the adenylatecyclase activity of the yeast membrane. PMID- 2990422 TI - Insertion of transcriptional elements outside the replication region can interfere with replication, maintenance, and stability of Co1E1-derived plasmids. PMID- 2990424 TI - Sex pheromones and plasmid transfer in Streptococcus faecalis: a pheromone, cAM373, which is also excreted by Staphylococcus aureus. PMID- 2990425 TI - General genetic recombination of bacterial plasmids. PMID- 2990423 TI - Mechanisms essential for stable inheritance of mini-F plasmid. AB - The F plasmid has its own partition mechanism and ccd mechanism (coupled cell division), besides its own replication mechanism, in order to be stably inherited into daughter cells through cell division. These 3 mechanisms are independent of one another. Therefore, when a DNA segment essential and sufficient for a mechanism is jointed to other heterologous plasmids, the segment is also functional. Most of natural low copy number plasmids might also have their own replication, partition, and ccd mechanisms. These 3 mechanisms may be fundamental to ensure stable inheritance for low copy-number replicons in prokaryotes. PMID- 2990426 TI - The origin of plasmid DNA transfer during bacterial conjugation. PMID- 2990427 TI - Studies on the transposition of IS1. PMID- 2990428 TI - Genetics of clindamycin resistance in Bacteroides. AB - Results presented in this paper have shown that a widely distributed LM resistance determinant is present on at least 3 distinct Bacteroides R-plasmids. In fact these plasmids bear no homology outside of the defined regions implicated in the LM resistance. This resistance determinant on pBF4, pBFTM10, and pBI136 is located within DNA segments bounded on each side by a directly repeated sequence of more than 500 bp. The intervening sequences of these 3 elements are variable, and range in size from about 3.7 kb to 7.2 kb (Fig. 7). Apart from the EcoRI/AvaI restriction sites which characterize the repeated sequence, there is a notable lack of common restriction sites within these elements. These results suggest that the elements do possess a certain degree of structural similarity but significant evolutionary divergence has occurred. The presence and location of the directly repeated sequences, their association with specific deletions, and their association with an antibiotic-resistance determinant, are features common to many antibiotic-resistance transposons described for other prokaryotes. In addition, these elements are highly mobile, being found on a number of R plasmids. The unique relationship between pBF4, pBI106, and pBI136 described here is a clear indication of the potential for these DNA sequences to move from one molecule to another. Given the extensive dissemination and the genetic and structural characteristics described above, it seems likely that the LM resistance determinant is carried on transposon-like elements present in pBF4, pBFTM10, and pBI136. However, further experimentation will be necessary to document the transposition event. Bacteroides strains such as B. fragilis V503, possess a transmissible LM-resistance determinant which does not appear to be associated with detectable extrachromosomal elements (5,9,10). Presently, a number of strains of this type have been found over a wide geographic area. The LM-resistance genes associated with these strains are apparently similar to the one carried on the Bacteroides R-plasmids because homology between the 2 has been observed. However, it is important to note that within the limits of Southern filter blot hybridization, neither V503 nor its transconjugants possess the directly repeated sequence found on the LM-resistance plasmids (Fig. 8). The elusive nature of the V503 LM-resistance elements presents an intriguing problem. One model that has been proposed is that these resistance determinants reside on a conjugative transposon similar to Tn916 of Streptococcus faecalis (3).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990429 TI - Effect of chromosome homology of plasmid transformation and plasmid conjugal transfer in Haemophilus influenzae. PMID- 2990430 TI - Conjugative sex plasmids of Streptomyces. PMID- 2990431 TI - The structure and source of plasmid DNA determine the cloning properties of vectors for Bacillus subtilis. PMID- 2990432 TI - Shuttle vector for Escherichia coli, Pseudomonas putida, and Pseudomonas aeruginosa. AB - A hybrid plasmid capable of replication in 2 different genera, Escherichia and Pseudomonas, was constructed. This plasmid DNA can be used as a cloning vector in E. coli and pseudomonades cells. The described hybrid plasmid pLD411 has been constructed on the basis of 2 small E. coli vector R-plasmids used in our laboratory and cryptic plasmid pWW2 or P. putida MT1. Plasmid pLD411 DNA was mapped with restrictases; its biological activity in transformations of different bacterial strains was studied, and the characteristics of transformed cells were also described. PMID- 2990433 TI - Plasmids in the degradation of chlorinated aromatic compounds. PMID- 2990434 TI - Notes on metabolic plasmid organization and expression. PMID- 2990436 TI - The plasmid-specified aerobactin iron uptake system of Escherichia coli. PMID- 2990435 TI - The structure of the mer operon. AB - The DNA sequence has been determined for a 3.8-kb region which encodes the mercury-resistance (mer) operon of the IncFII plasmid NR1. The sequence reveals 4 open reading frames which could encode proteins of 12,522, 9,429, 14,965, and 58,912 d corresponding to the 4 previously described Hg-inducible proteins detected in minicells carrying mer+ plasmids. The Hg(II) reductase protein sequence is about 90% homologous to that of Tn501, but the DNA sequence shows a homology of 60-70% to that of Tn501 except for short regions of very high homology. The entire mer region is 63.4% G-C overall. The region encoding the merR (positive regulatory) function has 3 possible open reading frames, 2 of which overlap in one direction and the third of which reads in the opposite direction. Attempts to visualize the polypeptide(s) encoded by the merR cistron were unsuccessful. PMID- 2990437 TI - Plasmid-mediated iron sequestering systems in pathogenic strains of Vibrio anguillarum and Escherichia coli. PMID- 2990439 TI - On the transposition and evolution of Tn1721 and its relatives. PMID- 2990438 TI - Cloning of an enzymatically active segment of the exotoxin-A gene of Pseudomonas aeruginosa. PMID- 2990440 TI - Structure, function, and regulation of the plasmid-encoded hemolysin determinant of Escherichia coli. PMID- 2990441 TI - Repeated DNA sequences recombine 1,000 times more frequently in a plasmid than in the chromosome of Bacillus subtilis. PMID- 2990442 TI - Retroviral oncogenes and human neoplasia. PMID- 2990443 TI - Structural basis for restriction-site polymorphism at the albumin locus in inbred strains of rats. AB - Two types of variant EcoRI restriction enzyme patterns of albumin-gene DNA fragments are found in different inbred strains of rats and reflect allelic polymorphism. The structural basis of the two allelic forms has been analyzed by mapping the EcoRI fragments using cloned albumin cDNA probes corresponding to the 5' or 3' end of the rat albumin mRNA and different genomic subclones. Additional restriction fragment length polymorphism has been detected using the restriction endonucleases HindIII and MspI. The results suggest that the two allelic variants differ from each other by multiple cleavage-site variations (base-pair substitutions) and by an insertion or deletion of DNA sequences. An extensive DNA sequence variation appears to exist between the two forms of the albumin gene; we have estimated that as much as 4% of the nucleotides in this region varied between the two alleles. All of this genetic variation is found in the intervening sequences of the gene and has no phenotypic manifestation. PMID- 2990444 TI - p-Cresol methylhydroxylase. Assay and general properties. AB - p-Cresol methylhydroxylase from Pseudomonas putida, an anaerobic dehydrogenase that catalyses the oxidation of p-cresol to p-hydroxybenzyl alcohol and then to p hydroxybenzaldehyde, is an enzyme of great interest in several respects. One of these is the fact that its flavoprotein and cytochrome c subunits may be reversibly dissociated with ease, with full regeneration of the activity and its native properties on recombining the components. Bisubstrate kinetic analysis of the unresolved enzyme gives parallel-line kinetics in double-reciprocal plots, whereas the reaction of the separated flavoprotein subunit with substrates is described by converging lines. The mechanistic implications of these behaviours are discussed. Reductive titration with dithionite results in the uptake of 3 electrons by the enzyme, with the intermediate formation of the anionic flavin radical [McIntire, Edmondson, Hopper & Singer (1981) Biochemistry 20, 3068-3075]. Reductive titration with substrates resulted initially only in reduction of the cytochrome subunit, followed by formation of the anionic radical and finally the fully reduced enzyme. These observations suggest rapid intermolecular electron transfer between p-cresol methylhydroxylase molecules. This paper also examines the effect of pH and ionic strength on the activity and specificity of the enzyme with respect to substrates and natural, as well as artificial, electron acceptors. The absorption coefficients of the enzyme and of its subunits in various oxidation states are also presented. PMID- 2990445 TI - Laser-flash-photolysis studies of p-cresol methylhydroxylase. Electron-transfer properties of the flavin and haem components. AB - p-Cresol methylhydroxylase, a heterodimer consisting of one flavoprotein subunit and one cytochrome c subunit, may be resolved into its subunits, and the holoenzyme may then be fully reconstituted from the pure subunits. In the present study we have characterized the reduction kinetics of the intact enzyme and its subunits, by using exogenous 5-deazariboflavin semiquinone radical generated in the presence of EDTA by the laser-flash-photolysis technique. Under anaerobic conditions the 5-deazariboflavin semiquinone radical reacts rapidly with the native enzyme with a rate constant approaching that of a diffusion-controlled reaction (k = 2.8 X 10(9) M-1 X s-1). Time-resolved difference spectra at pH 7.6 indicate that both flavin and haem are reduced initially by the deazariboflavin semiquinone radical, followed by an additional slower intramolecular electron transfer (k = 220 s-1) from the endogenous neutral flavin semiquinone radical to the oxidized haem moiety of the native enzyme. During the steady-state photochemical titration of the native enzyme at pH 7.6 with deazariboflavin semiquinone radical generated by light-irradiation the haem appeared to be reduced before the protein-bound flavin and was followed by the formation of the protein-bound anionic flavin radical. This result suggests that the redox potential of the haem is higher than that of the flavin, and that deprotonation of the flavin neutral radical occurred during the photochemical titration. Reduction kinetics of the flavoprotein and cytochrome subunits were also investigated by laser-flash photolysis. The protein-bound flavin of the isolated flavin subunit was reduced rapidly by the deazariboflavin semiquinone radical (k = 2.2 X 10(9) M-1 X s-1), as was the haem of the pure cytochrome c subunit (k = 3.7 X 10(9) M-1 X s-1). Flash-induced difference spectra obtained for the flavoprotein and cytochrome subunits at pH 7.6 were consistent with the formation of neutral flavin semiquinone radical and reduced haem, respectively. Investigation of the kinetic properties of the neutral flavin semiquinone radical of the flavoprotein subunit at pH 7.6 and at longer times (up to 5s) were consistent with a slow first-order deprotonation reaction (k = 1 s-1) of the neutral radical to its anionic form. PMID- 2990446 TI - Evidence that non-covalent forces, thiol and disulphide groups affect the structure and binding properties of the prolactin receptor on hepatocytes from pregnant rats. AB - Incubation of hepatocytes from pregnant rats with dithiothreitol decreased specific 125I-prolactin (125I-prl) binding to such cells by about 20% relative to control. This was not due to a non-specific effect of dithiothreitol on the cell membrane, since reduction also altered the binding of prl to solubilized partially purified receptor. Exposure of hepatocytes to N-ethylmaleimide (6 mM) for periods as brief as 1 min decreased the subsequent specific binding of 125I prl by more than 50%. N-Ethylmaleimide was less effective as an inhibitor of binding when applied after hepatocytes had been exposed to 125I-prl, binding being decreased by about 15%. Scatchard analysis demonstrated that the effect of N-ethylmaleimide resulted from loss of receptor-binding capacity without any substantial effect on the affinity of the prl receptor for hormone. Dithiothreitol diminished the affinity of lactogenic sites for prolactin without altering cellular binding capacity. These observations suggest that thiol and disulphide groups are present in the prl receptor and that these functional moieties regulate the formation and properties of prl receptor complexes. The species to which 125I-prl had bound were identified by affinity labelling. 125I prl was covalently coupled into saturable complexes of Mr 65000 and 50000. 125I human growth hormone (125I-hGH) was covalently incorporated into complexes of Mr 300 000, 220 000, 130 000, 65 000 and 50 000. Bovine growth hormone (bGH), but not prl, competed for 125I-hGH uptake into the 300 000-, 220 000- and 130 000-Mr complexes, indicating that these species were somatogenic. Prl, but not bGH, inhibited 125I-hGH uptake into 65 000- and 50 000-Mr complexes. This demonstrated that 125I-hGH in the presence of bGH could affinity-label lactogenic receptors. 125I-prl aggregates in Triton X-100, whereas 125I-hGH does not. Therefore lactogenic complexes to which 125I-hGH was bound in the presence of excess bGH were solubilized in Triton X-100 and characterized sequentially by gel filtration and affinity labelling. Prl receptors were eluted from columns of Sepharose 6B as a species of Mr380 000. Fractionation of the 380 000-Mr species on sodium dodecyl sulphate polyacrylamide gels resulted in the isolation of complexes of Mr 65 000 and 50 000. Thus non-covalent forces stabilize aggregates of the monomeric prolactin receptor. PMID- 2990447 TI - The energetics of early platelet responses. Energy consumption during shape change and aggregation with special reference to protein phosphorylation and the polyphosphoinositide cycle. AB - Among the different platelet responses, secretion requires the greatest amount of metabolic energy. The velocities of dense, alpha- and acid hydrolase granule secretion vary in parallel with the increase in energy consumption seen in thrombin-stimulated cells. This covariance is preceded by a phase in which energy consumption is increased without the extracellular appearance of secretion markers. By treating the platelets with thrombin and hirudin we have stimulated the platelets for short intervals and succeeded in separating shape change, single platelet disappearance and secretion to a great extent. In this report we show that the early increase in energy consumption reflects the energy requirement of aggregation but not of shape change. The cost of 100% of single platelet disappearance is 2.8 mumol of ATPeq. X (10(11) platelets)-1. Concurrent analysis of phosphorylation of Mr 20 000 and 47 000 proteins and of 32P-labelled phosphatidylinositol metabolites led to the following observations. Firstly, shape change is neither accompanied by an increase in protein phosphorylation nor by changes in the steady state levels of 32P-labelled phosphatidylinositol metabolites. Secondly, when aggregation occurs both proteins are phosphorylated, but the phosphatidylinositol metabolites do not change. Thirdly, when secretion follows, more phosphorylation of the Mr 47 000 protein occurs and initially only phosphatidic acid accumulates. At a later stage of the secretion responses, more protein phosphorylation and phosphatidic acid accumulation become evident, and are now accompanied by alterations in the steady state levels of 32P-labelled (poly)phosphoinositides. Hence, the early increase in energy consumption coincides with protein phosphorylation and, at a later stage, with alterations in (poly)phosphoinositides metabolites. This demonstrates that metabolic energy is directly involved in stimulus-response coupling in aggregating platelets. PMID- 2990449 TI - Effect of the chaotropic anions thiocyanate and perchlorate on the entry of ricin into Vero cells. AB - The effect of different anions on the sensitivity of Vero cells to ricin was investigated. The cells were equally sensitive to ricin when NaCl was replaced by NaBr, NaI, Na2SO4 or with iso-osmotic concentrations of mannitol. In contrast, NaSCN and NaClO4 strongly protected against ricin at pH 7.2, but not at pH 7.6. The possibility that the protective effect is due to the ability of chaotropic anions to decrease the pH close to the membrane is discussed. PMID- 2990448 TI - Inhibition of hydroxyapatite formation in collagen gels by chondroitin sulphate. AB - Crystal growth in native collagen gels has been used to determine the role of extracellular matrix macromolecules in biological calcification phenomena. In this system, type I collagen gels containing sodium phosphate and buffered at pH 7.4 are overlayed with a solution containing CaCl2. Crystals form in the collagen gel adjacent to the gel-solution interface. Conditions were determined which permit the growth of crystals of hydroxyapatite [Ca10(PO4)6(OH)2]. At a Ca/P molar ratio of 2:1, the minimum concentrations of calcium and phosphate necessary for precipitation of hydroxyapatite are 10 mM and 5 mM, respectively. Under these conditions, precipitation is initiated at 18-24h, and is maximal between 24h and 6 days. Addition of high concentrations of chondroitin 4-sulphate inhibits the formation of hydroxyapatite in collagen gels; initiation of precipitation is delayed, and the final (equilibrium) amount of precipitation is decreased. Inhibition of hydroxyapatite formation requires concentrations of chondroitin sulphate higher than those required to inhibit calcium pyrophosphate crystal formation. PMID- 2990450 TI - Collagenase inhibitory activity in human breast milk. PMID- 2990451 TI - Modification of opioid ligand binding in the central and the peripheral nervous system by different buffers. AB - The modification of binding parameters (equilibrium dissociation constant and binding capacity) of three opioid ligands (DADLE, Etorphine and EKC) on bovine adrenal medulla and rat brain membranes have been examined in three buffer systems: Tris-HCl 50 mM, Hepes-NaOH 10 mM and Tes-KOH 10 mM. Major differences of these parameters have been found: Hepes-NaOH provoked a diminution of the apparent number of binding sites, while a concomitant diminution of the KD and Bmax was observed in Tes-KOH buffer. Substitution of counterions in these two buffers produced further changes of binding characteristics: in Hepes buffer we have observed an abolition of 3H DADLE binding, an enhancement of 3H EKC binding and no modification of 3H etorphine binding characteristics. On the contrary an abolition of the specific binding of all three ligands in Tes buffer was found in the bovine adrenal medulla while minor changes were observed in rat brain. It is concluded that, inspite same disadvantages (substitution for bivalent cations and temperature dependence), Tris-HCl is the buffer of choice for the analysis of opioid binding site interactions. PMID- 2990452 TI - Epidermal growth factor stimulates diacylglycerol kinase in isolated plasma membrane vesicles from A431 cells. AB - We have examined the effect of epidermal growth factor(EGF) on three kinds of kinases activities, phosphatidylinositol(PI) kinase, phosphatidylinositol 4 phosphate[PI(4)P] kinase and diacylglycerol(DG) kinase that make important roles in the regulation of inositol phospholipids metabolism. When isolated plasma membrane vesicles from A431 cells were incubated at 30 degrees C with [gamma 32P]ATP and exogenously added DG, EGF enhanced the activity of DG kinase approximately 2-fold. This stimulation is found to be dose-dependent with a half maximal activation at 1 nM. In this case, EGF increased Vmax without changing Km Value for ATP or DG. Although this activation was observed in the absence of detergent, it was more evident when membrane vesicles were treated with 1 mM deoxycholate. Interestingly, the effect of EGF was only detected in magnesium containing medium. The use of manganese instead of magnesium diminished the stimulatory effect in either condition, presence or absence of deoxycholate. On the other hand, the stimulation of PI kinase or PI(4)P kinase activity was not caused by EGF. These results suggest that DG kinase activation by EGF makes important roles in cellular responses leading to cell growth. PMID- 2990453 TI - Role of phospholipids in the activation of the Ca2+-dependent neutral proteinase of human erythrocytes. AB - Activation of the Ca2+-dependent neutral proteinase of human erythrocytes in the presence of Ca2+ and a digestible substrate (Pontremoli, S., Sparatore, B., Melloni, E., Michetti, M. and Horecker, B.L. 1984, Biochem. Biophys. Res. Communs. 123, 331-337) is promoted by phospholipids such as phosphatidylcholine, phosphatidylinositol and phosphatidylserine. The presence of at least one unsaturated fatty acid chain is essential and metabolic derivatives such as dioleylglycerol, phosphorylserine and free fatty acids are ineffective. The most effective promoter was a freshly prepared mixture of phospholipids from human erythrocyte membranes. Activation involves conversion of the 80 kDa proenzyme (procalpain) subunit to the 75 kDa active proteinase and is irreversible. Phospholipids act by producing a large decrease in the concentration of Ca2+ required for the conversion of procalpain to active calpain. PMID- 2990454 TI - Tumor promoters enhance basal and growth hormone releasing factor stimulated cyclic AMP levels in anterior pituitary cells. AB - Tumor promoters, such as phorbol esters and teleocidin, amplified the ability of growth hormone releasing factor to increase pituitary cyclic AMP levels. This effect of tumor promoters was concentration-dependent, could be observed in 5 minutes, and was over by 4 hours. Inactive tumor promoters (i.e., 4-alpha didecanoate) had no effect on this system, whereas a synthetic diacylglycerol (i.e., 1-oleoly-2-acetyl glycerol), mimicked the action of tumor promoters. Due to the known stimulation of protein kinase C by both tumor promoters and diacylglycerols, we suggest that this calcium and phospholipid dependent protein kinase C can enhance the ability of the growth hormone releasing factor receptor to activate the cyclic AMP generating system. PMID- 2990455 TI - Deoxycholate induces the preferential hydrolysis of polyphosphoinositides by human platelet and rat corneal phospholipase C. AB - Deoxycholate promotes phospholipase C degradation of endogenous phosphatidyl[3H]inositol (Pl), phosphatidyl[3H]inositol monophosphate (PIP) and phosphatidyl[3H]inositol bisphosphate (PIP2) in rat cornea and human platelets. Hydrolysis of phosphatidyl[3H]inositol significantly lags polyphospho[3H]inositide degradation. Concomitantly, formation of [3H]inositol monophosphate (IP1) lags behind [3H]inositol bisphosphate (IP2) and [3H]inositol trisphosphate (IP3) production. These results demonstrate that rat cornea and human platelet phospholipase C cause a preferential hydrolysis of the endogenous polyphosphoinositides rather than phosphatidylinositol. PMID- 2990456 TI - A tumor promoter enhances the phosphorylation of polyphosphoinositides while decreasing phosphatidylinositol labelling in lymphocytes. AB - 12-O-Tetradecanoyl-phorbol-13-acetate, a comitogen for lymphocytes, suppresses the concanavalin A-induced accumulation of 3 2P-phosphatidyl-inositol, in mouse spleen lymphocytes incubated with 3 2P-orthophosphate. The comitogenic tumor promoter does not affect the rate of de novo synthesis of phosphatidylinositol, as measured by [3H]-glycerol incorporation. Mitogen stimulates the incorporation of [3 2P]-phosphate into phosphatidylinositol, phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-bisphosphate, but the tumor promoter suppresses only the increased labelling of the phosphatidylinositol, while it enhances phosphorylation of the polyphosphoinositides. PMID- 2990457 TI - Free radicals of tocopherol model compound, 6-hydroxy-2,2,5,7,8 pentamethylchroman which are produced from the reaction with superoxide ion, O2: studies by high-performance liquid chromatography. AB - Reaction of superoxide ion, O2-, with alpha-tocopherol model compound, 6-hydroxy 2,2,5,7,8-pentamethylchroman (lb), was investigated by high-performance liquid chromatography (HPLC). Chromatogram of the reaction mixture showed three peaks with retention times of 2.5, 1.8 and 1.5 min, and each peak height was dependent on the concentration of O2. Chemical species having the retention time of 1.5 min was ascribed to chromanoxyl radical (3), and the other chemical species having the retention times of 2.5 and 1.8 min were identified with the model compound (lb) and 2-hydroxy-2-methyl-4-(3, 5, 6-tri-methylbenzoquinone-2-yl) butane (2), respectively. This is a first evidence that the free radicals from tocopherol model compounds was separated by HPLC. PMID- 2990458 TI - Interaction of cardiodigin, endogenous inhibitor of Na+,K+-ATPase, with antidigoxin and antidigitoxin antibodies. AB - The present study was undertaken to further characterize the immunoreactivity of cardiodigin, digitalis-like factor present in mammalian tissues. Guinea-pig heart extracts purified by reverse-phase low pressure and high pressure liquid chromatographies were analysed for their cross-reactivity with antidigitoxin and antidigoxin antibodies. The putative digitalis-like factor showed an affinity about 10 times higher for antidigoxin antibodies than for antidigitoxin antibodies. EC50 ratios (digoxin/digitoxin assay) determined at two purification steps were different from those of digoxin and digitoxin. These results show that cardiodigin presents molecular determinants recognizable by antidigoxin and antidigitoxin antibodies but that it is a chemical entity different from these well known cardioactive steroids. PMID- 2990459 TI - Effects of LTB4 and Ca++ ionophore A23187 on the release by human alveolar macrophages of factors controlling fibroblast functions. AB - Cultured human alveolar macrophages from smokers with lung cancer produced spontaneously variable amounts of factors stimulating fibroblast proliferation and production of prostaglandin E2 and collagenase by fibroblasts. These biological activities belong to molecules similar or identical to interleukin 1. Exogenous leukotriene B4 added to alveolar macrophage cultures increased the production of these factors. The Ca++ ionophore A23187 was found to have similar effects. By the control of monokine production, leukotriene B4 locally released by inflammatory cells may modulate lung fibroblast functions. PMID- 2990460 TI - Reconstitution of phosphate-linked antiport from Streptococcus lactis. AB - Membrane protein solubilized by octyl-beta-D-glucopyranoside in the presence of dispersed phospholipid was incubated with bath-sonicated liposomes and additional detergent. The proteoliposomes formed on dilution showed transport and exchange properties consistent with a reconstitution of phosphate:sugar 6-phosphate antiport. Thus, phosphate self-exchange was found only when protein from induced cells was used; this exchange was blocked by a sugar 6-phosphate, not by a sugar 1-phosphate; and proteoliposomes supported an accumulation of 2-deoxyglucose 6 phosphate with no added source of energy. Solubilization and reconstitution of protein was most effective when performed in the presence of gram-positive phospholipids. PMID- 2990461 TI - Electric field mediated transformation: isolation and characterization of a TK+ subclone. AB - Transformation of mammalian TK- cells by a plasmid carrying the TK gene from Herpes virus simplex 1 (pAGO) was mediated by electroporation. The cells were treated either in suspension or growing in monolayers directly in the petri dish. The yield of transformation was between 8.10(-5) and 2.10(-4) per microgram DNA depending on the experimental conditions. The structure of the integrated DNA was investigated proving the occurrence of a duplication process that affected preferentially the pBR322 part of the pAGO DNA (60 copies per cell). The TK gene that gave the TK+ phenotype to the selected clone was present in less than 6 copies. PMID- 2990463 TI - Hybrid forms of phosphoglycerate mutase and 2,3-bisphosphoglycerate synthase phosphatase. AB - Purified phosphoglycerate mutase from pig skeletal muscle and 2,3 bisphosphoglycerate synthase-phosphatase from pig erythrocytes were hybridized "in vitro". The hybrid showed a behaviour on electrophoresis and on ion-exchange chromatography similar to that of a naturally occurring enzyme with phosphoglycerate mutase, 2,3-bisphosphoglycerate synthase and 2,3 bisphosphoglycerate phosphatase activities present in pig skeletal and heart muscle. Both the hybrid and the muscle enzyme possess similar activities ratio. From these and previous data it is suggested that the six enzymatic forms with phosphoglycerate mutase, 2,3-bisphosphoglycerate synthase and 2,3 bisphosphoglycerate phosphatase activities detected in mammalian tissues (Carreras et al. 1981, Comp. Biochem. Physiol. 70B, 477-485) result from combination of three subunits (types M, B and E). PMID- 2990462 TI - Platelet-activating factor mediated effects on human neutrophil function are inhibited by pertussis toxin. AB - Pretreatment of human neutrophils with pertussis toxin inhibits platelet activating factor-mediated chemotaxis, superoxide generation, aggregation, and release of lysozyme. By contrast, superoxide generation observed in the presence of phorbol-12-myristate-13 acetate is unaffected. Our results suggest that a target protein for pertussis toxin, probably a GTP binding protein termed "Ni", is involved in the actions of platelet-activating factor on human neutrophils. PMID- 2990464 TI - Adenylate cyclase activity and cyclic AMP production in the outer and inner zones of the adrenal cortex. AB - It has been reported that cells isolated from the inner zone of the guinea pig adrenal cortex fail to have a steroidogenic response to ACTH. To further explore this, adenylate cyclase activity of membrane particles and cAMP production by cells prepared from the inner and outer adrenocortical zones were determined. The cAMP response to ACTH and forskolin was similar for cells from both zones. Basal adenylate cyclase activity was significantly higher in the inner zone; and while absolute responses to ACTH, GppNHp, GTP, NaF, and forskolin were greater for the inner zone, relative responses were similar for the two zones. These observations suggest that the inner zone of the guinea pig adrenal cortex may have a defect in ACTH action at a step(s) beyond cAMP formation. PMID- 2990465 TI - Localization of the human prealbumin gene to chromosome 18. AB - A human liver cDNA library was screened using an oligonucleotide probe based on the amino acid sequence of human prealbumin. The cDNA insert of one positive clone was sequenced and found to contain the entire coding sequence of human prealbumin plus untranslated 5' and 3' regions. This cDNA was used to probe DNA from a panel of mouse/human somatic cell hybrids. Only those hybrids containing human chromosome 18 showed the human-specific hybridization pattern, thereby localizing the human prealbumin gene to this chromosome. PMID- 2990466 TI - Specific receptors for atrial natriuretic polypeptide on basolateral membranes isolated from rat renal cortex. AB - The binding of alpha-human atrial natriuretic polypeptide (alpha-hANP) to brush border and basolateral membranes isolated from the rat renal cortex was studied at 0 degree C by a rapid filtration technique. Specific binding of 125I-alpha hANP to basolateral membranes reached a steady state at 4 hr. The binding to brush border membranes was maximal at 5-15 min and then rapidly decreased. The analysis of incubation mixtures with basolateral membranes revealed little degradation of 125I-alpha-hANP during the 4-hr incubation, while there was extensive degradation of the ligand with brush border membranes during the 30-min incubation. High affinity binding of 125I-alpha-hANP was demonstrated on basolateral membranes but not on brush border membranes. These data suggest that specific receptors for alpha-hANP are localized on basolateral membranes of the renal cortex. PMID- 2990467 TI - Characterization of insulin receptor carbohydrate by comparison of chemical and enzymatic deglycosylation. AB - To characterize the carbohydrate moieties of the insulin receptor on IM-9 lymphocytes, the cells were surface iodinated and solubilized, and the insulin receptors were precipitated with anti-receptor antibody. The precipitates were resuspended, subjected to either enzymatic digestion or chemical treatment with trifluoromethanesulfonic acid and the relative mobilities of the alpha and beta subunits before and after treatment were analyzed by polyacrylamide gel electrophoresis and autoradiography. The results indicate that the alpha subunit possesses primarily N-linked carbohydrate which is both complex (Endoglycosidase F sensitive) and polymannose (Endoglycosidase H sensitive). The beta subunit also contains polymannose oligosaccharide units and has, in addition, a substantial amount of carbohydrate which is removed by chemical treatment but is not susceptible to Endoglycosidase F, suggesting the presence of O-linked saccharides. The apparent molecular weights of the core protein of the mature alpha and beta subunits as determined by gel electrophoresis following complete deglycosylation are 98 kDa and 80 kDa, respectively. PMID- 2990468 TI - Characterization of restriction endonuclease maps of hepatitis B viral DNAs. AB - The HBV DNA isolated from Dane particles of 9 patients' plasma was cloned into the EcoRI or BamHI site of the pUC8 plasmids. Two plasmids with full length HBV DNA and four plasmids containing the HBV surface antigen gene were obtained. Based on our cloned HBV DNA and a comparison with 7 complete sequences and 5 restriction endonuclease patterns of HBV DNA published by others, we can recognize common restriction sites shared by different subtypes (adw, adr, ayw, and adyw): (1) a HincII site in the S gene, (2) a BamHI site in the X region, and (3) two BglII sites in the C gene. In addition adw has specific sites for HincII, BamHI, and PstI in the pre-S region. A unique XhoI site is present in the pre-S region in all subtypes except for adw. PMID- 2990469 TI - Characterization of calcium-activated neutral protease (CANP)-associated protein kinase from bovine brain and its phosphorylation of neurofilaments. AB - Calcium-activated neutral protease with low affinity for calcium (CANP II, Mr 76,000) can be purified to apparent homogeneity by casein affinity chromatography but contains cyclic-AMP dependent protein kinase activity. CANP II-associated kinase from bovine brain copurifies with protease activity through multiple chromatographic procedures but can be separated by cyclic-AMP affinity chromatography. Isolated protein kinase has subunits of Mr 80,000, 53,000 and 42,000. The kinase preferentially "autophosphorylates" CANP II, but histones, phosphorylase b and neurofilament proteins are also good substrates. The concentrations for half-maximal phosphorylation activity (Km) of cyclic-AMP, (32P)ATP and Mr 150,000 neurofilament protein substrate are 0.2, 6.0 and 0.5 microM, respectively. The specific activity of CANP II associated kinase in phosphorylating neurofilament proteins is intermediate between that of neurofilament- and MAPs 2-associated kinases. PMID- 2990470 TI - Atrial natriuretic peptides stimulate renal gluconeogenesis. AB - Atrial natriuretic peptide (5-28AA; ANP) and atrial extract (ANS) stimulated rat renal gluconeogenesis in cortical tubule suspension in a dose dependent fashion only from substrates that enter gluconeogenesis via phosphoenol-pyruvate carboxylase. The effects of ANP and ANS were significantly potentiated by cAMP and cGMP, whereas methoxamine showed no effect. Extracellular calcium revealed a key role for ANP and ANS response to gluconeogenesis: a concentration of calcium higher than 1 mM was essential. Isolated cells from cortex which lost cell membrane polarity by warming but responded solely to cAMP and cGMP showed no effect by ANP nor ANS. These data suggest that ANP or ANS may act mainly from the basolateral site in the proximal tubule cell and promote gluconeogenesis through cAMP and/or cGMP system. PMID- 2990471 TI - Bovine heart cytochrome c oxidase preparations contain high affinity binding sites for magnesium as well as for zinc, copper, and heme iron. AB - Metal contents of six bovine heart cytochrome c oxidase preparations were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Magnesium is present in consistent amounts reflected by Fe/Mg and Mg/Zn ratios of 2.06 +/- 0.18 and 1.10 +/- 0.14, respectively. More copper than iron is always present; the average Cu/Fe atom ratio is 1.27 +/- 0.10. Calcium is found in significant but variable amounts. Mg, Zn, Fe, and Cu are each bound with high affinity as shown by a resistance to removal upon dialysis against various media. The Cu in excess of Fe, the Mg, and the Zn may each have catalytic and/or structural roles in the oxidase. The observed metal stoichiometry suggests that a dimeric catalytic unit with 5 Cu, 4 Fe, 2 Zn, and 2 Mg may be present in the inner mitochondrial membrane to carry out O2 reduction and H+ pumping. PMID- 2990472 TI - Modulation of the activity of the platelet-derived growth factor receptor by phorbol myristate acetate. AB - Phorbol myristate acetate (PMA) weakly activates Na+/H+ exchange in NR-6 cells. Simultaneously, PMA blocks the activation of Na+/H+ exchange by platelet-derived growth factor or by serum. Phorbol esters that do not activate protein kinase C do not show this metabolic response. We conclude that activation of Na+/H+ exchange by platelet-derived growth factor or serum does not require the intermediate activation of protein kinase C. We postulate from this and previous observations that a major role of protein kinase C is to act as an inhibitor of the activity of cell surface receptors, in particular mitogen receptors. PMID- 2990473 TI - Activation of human platelets by ADP causes a rapid rise in cytosolic free calcium without hydrolysis of phosphatidylinositol-4,5-bisphosphate. AB - Phosphatidylinositol-4,5-bisphosphate decreased 40% within 10 seconds after the addition of thrombin to platelets. This thrombin-induced loss was accompanied by a corresponding increase of inositol phosphates. In contrast, within the first 60 seconds after exposure of platelets to ADP there was no detectable change in the amounts of phosphatidylinositol-4,5-bisphosphate or inositol phosphates. Both thrombin and ADP, however, caused a very rapid rise of cytosolic free calcium, as measured by Quin-2. The magnitude of this rise of calcium was similar for the two agonists. These results suggest that in platelets, agonist stimulation may lead to increased cytosolic free calcium independently of phosphatidylinositol-4,5 bisphosphate degradation. PMID- 2990474 TI - Receptor-mediated uptake of asialoganglioside liposomes: sub-cellular distribution of the liposomal marker in isolated liver cell types. AB - The use of asialo GM1-containing small unilamellar liposome preparations in vivo caused a 2.8-fold increase in the uptake by the liver as compared with the control (neutral) preparations (without asialo GM1). The uptake of negatively charged dicetylphosphate and dipalmitoyl phosphatidic acid-containing small unilamellar liposomes was found to be 1.6-and 1.8-fold respectively higher than that of the neutral preparations. In studies with isolated liver cell types, inhibition of the galactosylated liposome uptake by asialofetuin indicated a possible involvement of hepatic galactose receptors in the recognition of asialo GM1 liposomes by the hepatic parenchymal cells, which in turn were found to be mainly responsible for the enhanced incorporation of these liposomes in the liver. Sub-cellular distribution studies with isolated liver cell types indicated lysosomal localization of the liposomes both in parenchymal and nonparenchymal cells, and it has been proposed that the asialo GM1 liposomes are cointernalized with asialofetuin through a common lysosomal route of ligand internalization. PMID- 2990475 TI - Gluconate kinase from Zymomonas mobilis: isolation and characteristics. AB - The enzyme gluconate kinase EC 2.7.1.12 has been found at high levels in glucose grown Zymomonas mobilis cells. A simple procedure, based on differential dye ligand chromatography, has been used to isolate the enzyme, purifying it some 600 fold. The purified enzyme is a monomer of molecular weight 18,000 Da, which is much smaller than other gluconate kinases reported. It has a relatively low affinity for ATP. (Km = 1.5 mM), but high for gluconate (Km = 0.33 mM), and has little activity with any other potential substrates. PMID- 2990476 TI - The effect of tetrodotoxin on mammalian sarcolemmal proteins and glycoproteins. AB - This study evaluates the relative contributions of neural impulse activity and neurotrophic (non-impulse) factors by comparing the effects of denervation and the blockage of neural impulse activity on various biochemical parameters of mixed muscle sarcolemma. Muscle paralysis was induced by repeated injection of Tetrodotoxin (TTx) to the sciatic nerve. After seven days of inactivity the isolated sarcolemma was analyzed for protein and glycoprotein composition, Na+/K+ ATPase activity, Concanavalin A binding to intact sarcolemma and to carbohydrate components separated in SDS-polyacrylamide gels and sialyl and galactosyl transferase activity. The results have shown that following muscle inactivity all of the parameters except glycosyl transferases changed in a similar manner but to a lesser degree than denervation. It is concluded that trophic factors in addition to neural impulse activity play a role in the regulation of a number of surface membrane properties. PMID- 2990477 TI - Occurrence of an ecto-phosphoprotein phosphatase in goat epididymal spermatozoa. AB - Intact spermatozoa from goat cauda epididymis possess phosphoprotein phosphatase activity that causes dephosphorylation of externally added [32p]histones. The enzymic reaction was linear with time for at least 15 min and there was little uptake of [32p]histones by these cells. The activity of the enzyme of the whole spermatozoa was not due to contamination of the broken cells or epididymal plasma and leakage of the intracellular enzymic activity during incubation. The activity of the phosphoprotein phosphatase was strongly inhibited by the thiol reagent: p chloromercuribenzenesulfonic acid, which is believed not to enter the cells. There was no appreciable loss of the enzymic activity from the cells when washed with EDTA (2.0 mM) or a hyperosmotic medium. These data are consistent with the view that the observed activity of the enzyme is located on the spermatozoal external surface. Studies with unlabelled p-nitrophenyl phosphate and beta glycerophosphate indicate that the sperm ecto-enzyme is not a non-specific phosphatase. PMID- 2990479 TI - ATPase activities in kidney basolateral plasma membranes of young and old rats. AB - The present work studied the turnover rate of (Na+ + K+)-ATPase as well as Mg2+- and Na+- ATPase activities in basolateral plasma membranes from kidney cortex cells of young and old rats. It was found that, as for the homogenates, the turnover rate of the (Na+ + K+)-ATPase was diminished by aging in about 40%. The Mg2+-ATPase activities on the other hand, were similar for the rat kidneys of young and old, in both the homogenates as well as the basolateral plasma membrane fractions. PMID- 2990478 TI - Relation of blood glucose levels to the changes in plasma levels of various hydrolytic enzymes in diabetic patients. AB - Plasma levels of various hydrolytic enzymes in diabetic patients, as diagnosed by their blood glucose levels, were compared with those of control subjects. Most of the levels of the aminopeptidase activities examined were significantly increased, while those of angiotensin-converting enzyme (ACE) and esterase decreased, in diabetic patients. The results of multivariate analysis suggest that the increased aminopeptidase activities are related rather to secondary organ lesions than to the primary pancreatic lesion. It seems that the blood glucose level is not necessarily a good indicator of metabolic abnormalities underlying this pathological condition. PMID- 2990480 TI - A possible action of nicardipine on the cardiac sarcolemmal Na+-Ca2+ exchange. AB - The effects of nicardipine on sodium-calcium exchange activity of cardiac sarcolemma-enriched vesicles isolated from the rat heart were examined. Sodium loaded, sarcolemma-enriched vesicles, when exposed to a medium containing 40 microM CaCl2, exhibited about 5 nmoles Ca2+/mg protein of the maximal calcium uptake; the initial rate was 21 nmoles Ca2+/mg protein/min. The calcium uptake was dependent on the extravesicular concentration of calcium ion. Nicardipine at concentrations of 0.1 to 10 microM depressed the rate of calcium uptake activity by 60-90%. The isolated membrane vesicles preloaded with Ca2+ showed a calcium efflux activity, when exposed to a medium containing sodium ion. The rate of calcium efflux was 2.5 nmoles Ca2+/mg protein/min, when measured in a medium containing 6.5 mM NaCl. The efflux rate was facilitated with increased concentrations of sodium ion in the medium. About 75% of the preloaded calcium in the vesicles was released within 3 min of incubation. The rate of calcium efflux was stimulated in the presence of 0.1 to 10 microM nicardipine (2.5- to 4-fold increase). The present results suggest a possible action of nicardipine on the sodium-calcium exchange mechanism at cardiac sarcolemmal sites. PMID- 2990481 TI - Inhibition of the gastric (H+ + K+)-ATPase by fenoctimine. AB - The effects of fenoctimine, an inhibitor of gastric acid secretion, on the microsomal (H+ + K+)-ATPase were studied. In the micromolar concentration range, fenoctimine inhibited hydrolysis of ATP and p-nitrophenyl phosphate by the (H+ + K+)-ATPase. Inhibition was reversible and noncompetitive with substrate. The apparent Ki was dependent on the concentration of membranes, being increased by added liposomes or high microsomal membrane concentrations. Over the concentration range that (H+ + K+)-ATPase was inhibited, fenoctimine increased the turbidity of microsomal suspensions. The effects of fenoctimine were not specific for the gastric (H+ + K+)-ATPase, since the hydrolytic activities of the (Na+ + K+)-ATPase and mitochondrial ATPase were also inhibited by the drug. These results suggest that inhibition of hydrolysis may not be the direct result of an interaction between the (H+ + K+)-ATPase and fenoctimine but the secondary effect of a fenoctimine-induced perturbation of the microsomal membrane. PMID- 2990482 TI - Mode of inhibitory action of melittin on Na+-K+-ATPase activity of the rat synaptic membrane. AB - The effects of melittin from bee venom, cardiotoxin from Formosan cobra venom, and ouabain on Na+-K+-ATPase activity of the synaptic membrane isolated from rat cerebral cortex were studied. Melittin was the most potent in inhibiting Na+-K+ ATPase activity. Mg2+-ATPase was less susceptible than Na+-K+-ATPase to the inhibitory action of toxins. High K+ (30 mM) reversed the inhibitory action of melittin on Na+-K+-ATPase but did not affect that of cardiotoxin. A comparison between the effects of ouabain and melittin was studied, using double-reciprocal plots of Na+-K+-ATPase activity against K+. It was shown that both were competitive with K+ for binding to the K+ site. Moreover, a median-effect plot revealed that ouabain and melittin antagonized each other when inhibiting Na+-K+ ATPase. Phosphatidylcholine (PC) was the only one of the phospholipids tested capable of protecting Na+-K+-ATPase from the inhibitory action of melittin but not that of ouabain. However, the inhibitory action of cardiotoxin on this enzyme was decreased by phosphatidylserine and sphingomyelin, in addition to PC. All of these findings suggest that the melittin polypeptide potently inhibits Na+-K+ ATPase, possibly by binding to the K+ site. PMID- 2990483 TI - Species difference in temperature dependence of cardiac (Na+ + K+)-ATPase activity. AB - Cardiac (Na+ + K+)-ATPase from several species of animals were comparatively studied with respect to their molecular form and temperature dependence. The molecular weight of the catalytic subunit varied a little among species, but the difference did not correlate with the sensitivity of the enzyme to ouabain inhibition. Analysis of Arrhenius plots of the activities of the enzymes showed that enzymes showing break points of 24-25 degrees were relatively insensitive to ouabain inhibition whereas those enzymes with break points of 29-31 degrees were much more sensitive to the glycoside. This suggests that there is a difference in the interaction of cardiac (Na+ + K+)-ATPase with lipids between the ouabain sensitive and -insensitive animals. PMID- 2990484 TI - Captopril removal by rabbit lung in vivo. AB - Removal of [14C]captopril by the lungs of anesthetized rabbits was measured by the multiple indicator dilution technique. After coinjection of indocyanine green (ICG) and [14C]captopril into the jugular vein of anesthetized rabbits, serial blood samples were collected from the carotid artery and each was analyzed for its content of both substances. Percent removal (R) of captopril after its initial injection of 10 nmoles captopril/kg (calculated at the peak of the ICG outflow curve) was 40.2 +/- 2.5 (S.E.M.) and was significantly greater than R after a second injection of 10 nmoles captopril/kg (20.1 +/- 2.4) 1 hr later. Removal of 70 nmoles captopril/kg (5.8 +/- 3.0 after first injection, 6.4 +/- 2.2 after second injection) was significantly lower than R of 10 nmoles captopril/kg. During a single pulmonary passage of either dose of captopril, R was inversely related to the calculated fractional concentration of intravascular captopril. Pulmonary metabolism of the angiotensin converting enzyme (ACE) substrate [3H]benzoyl-Phe-Ala-Pro [( 3H]BPAP) was 70.1 +/- 1.7% in the absence of captopril, and was reduced significantly to 27.4 +/- 2.4% by 10 nmoles captopril/kg and 7.6 +/- 0.2% by 6 mumoles BPAP/kg. BPAP (6.4 +/- 0.6 mumoles/kg) significantly reduced R of the first and second injections of 10 nmoles captopril/kg but this effect was selective, since BPAP did not reduce pulmonary removal of [14C]serotonin. These data indicate that pulmonary removal of captopril in vivo is saturable and may primarily reflect binding of the drug to pulmonary endothelial ACE. PMID- 2990485 TI - Effects of ascorbic acid on biologically obtained diaziquone free radicals. PMID- 2990486 TI - Microsomal calcium-accumulating ability of bovine coronary artery and aorta. AB - Calcium uptake and binding activities of microsomal fractions from bovine coronary artery and aorta were examined. The isolated microsomal fraction of the coronary artery and aorta showed 7- to 8-fold higher glucose-6-phosphatase activity and 4- to 6-fold higher NADPH-cytochrome c reductase activity as compared with the corresponding values for the homogenate fraction. Coronary artery and aorta microsomal calcium uptake activities were 118 and 159 nmoles Ca2+/mg protein/10 min in the presence of 100 microM CaCl2, respectively. These activities for bovine vascular smooth muscle microsomes are higher than those of other species investigated. The calcium uptake activities were dependent on calcium concentrations ranging from 5 to 50 microM in the assay medium. The onset of the reaction for aorta microsomal calcium uptake was faster than that for the coronary artery. The calcium uptake activity was also dependent on ATP, but it was practically independent of oxalate ions in the assay medium. Microsomal calcium binding activities of the coronary artery and aorta were maximal at 20 min of incubation under the present experimental conditions. A lower Km value of the aortic calcium binding for ATP was obtained as compared with that for the coronary artery. The present experiment explored several characteristics of the microsomal calcium-accumulating ability of vascular smooth muscle, which provides meaningful information for further study on cellular calcium movements in vascular smooth muscle. PMID- 2990487 TI - Inhibition of phosphatidylinositol phosphodiesterase activity in skeletal muscle by metal ions and drugs which block neuromuscular transmission. AB - The calcium-dependent phosphatidylinositol phosphodiesterase activity of skeletal muscle cytosol was determined. The enzyme was inhibited by Zn2+, Cu2+ and Pb2+ ions but Mg2+ and Mn2+ were without effect. The antimalarial drugs chloroquine and the quinine and the aminoglycoside antibiotics gentamicin and neomycin all of which, like Zn2+, have been shown to block neuromuscular transmission, also inhibited the enzyme. PMID- 2990488 TI - Inhibition of epipodophyllotoxin cytotoxicity by interference with topoisomerase mediated DNA cleavage. AB - This laboratory and others previously proposed that the antitumor effects of the epipodophyllotoxin compounds are based on their abilities to stimulate DNA cleavage by a DNA topoisomerase. To explore this relationship further, we studied the intercalating agent ethidium bromide and found that it blocked epipodophyllotoxin-induced DNA cleavage by DNA topoisomerase II in vitro as well as in vivo. Using an in vitro assay consisting of purified calf thymus DNA topoisomerase II, end-labeled DNA, and the epipodophyllotoxin teniposide, we found that ethidium bromide markedly interfered with the enzyme-mediated DNA cleavage. Furthermore, ethidium bromide also blocked the formation of DNA single- and double-strand breaks in mouse L1210 cells when exposed to the epipodophyllotoxin etoposide. This effect cannot be explained by alterations in drug accumulation since steady-state drug concentrations were unchanged, and the effect was also observed in isolated nuclei. In addition to its effects on epipodophyllotoxin-mediated DNA breakage, ethidium bromide also potently inhibited the cytotoxic effects of etoposide but only when present during drug treatment. Thus, we believe that ethidium bromide may be a useful tool to investigate drug-induced perturbations of topoisomerase activity and their relationship to antitumor effect. Our data strongly support the hypothesis that the antitumor activity of epipodophyllotoxins is based on the ability to stimulate the formation of a cleavable complex between DNA topoisomerase and DNA. PMID- 2990489 TI - Effects of guanethidine on electron transport and proton movements in rat heart, brain and liver mitochondria. AB - Guanethidine at 5-25 mM concentrations was found to induce up to 79% inhibition of ADP-stimulated (state III) oxygen consumption in isolated rat heart, brain or liver mitochondria, when the added substrate was glutamate or succinate, but the inhibition was considerably lower (24% or less) when respiration was supported by ascorbate plus tetramethylphenylenediamine (TMPD). Comparable results were seen regarding ADP-stimulated proton uptake, where even greater inhibition (up to 94% with glutamate or succinate, but not ascorbate plus TMPD) was found. Similar but somewhat less marked effects were also seen in resting (state IV) respiration and on the acceptor control ratio (state III/state IV respiration). 2,4-Dinitrophenol was unable to relieve guanethidine-induced inhibition of electron transport. These results indicate that guanethidine inhibits primarily mitochondrial electron transport itself, and that the site where such inhibition is more marked is located in the span between ubiquinone and cytochrome c of the respiratory chain. It is, therefore, suggested that active guanethidine uptake by noradrenergic neurons can lead to a high drug concentration in their cytoplasm and hence to mitochondrial alterations that can contribute to the pharmacological effect of this drug. Our results demonstrate the interaction between guanethidine and the electron transport chain of mitochondria derived from different tissues and, therefore, support this hypothesis. PMID- 2990490 TI - Ascorbic acid: an endogenous inhibitor of isolated Na+,K+-ATPase. AB - During attempts to isolate and identify an endogenous ligand for the glycoside binding sites on Na+,K+-ATPase, bovine adrenal glands were found to contain a potent inhibitor of isolated Na+,K+-ATPase. The inhibitory principle was extracted from adrenal cortex, following homogenization in NaHCO3 solution and separation on a Sephadex G-10 column. The active principle was recovered from a fraction which eluted from the column after the 3H2O peak. The extract inhibited isolated Na+,K+-ATPase and the specific [3H]ouabain binding reaction. Sensitivity of the enzyme to the inhibitory action of the extract was species and tissue dependent; however, the pattern and the magnitude of the sensitivity were different from those of the digitalis glycosides. Moreover, the inhibitory principle failed to inhibit sodium pump activity, estimated from ouabain inhibitable 86Rb+ uptake by guinea pig brain slices. The activity of the extract to inhibit isolated Na+,K+-ATPase was stable under acidic condition but was lost rapidly at neutral pH, and could be eliminated by EDTA. In an acidic medium, the inhibitory principle had an absorption maximum at 244 nm which shifted to 264 nm and decayed rapidly at neutral pH. By using mass spectrometry, the principle was identified to be ascorbic acid, which has been shown previously to inhibit isolated Na+,K+-ATPase under appropriate conditions. Because ascorbic acid was incapable of inhibiting the sodium pump in intact cells, this inhibitor of the isolated enzyme does not appear to be the endogenous ligand which regulates sodium pump activity in vivo. PMID- 2990491 TI - Comparative studies of some semisynthetic K-strophanthins with natural cardiac glycosides. AB - In this study the pharmacological effects of seven semisynthetic cardenolides have been investigated and compared with those of 11 natural cardiac glycosides. These compounds are of different potency on electrically driven isolated guinea pig atria. Their concentration response curves showed different slopes, which could be an indication of varying therapeutic range. The pharmacodynamics of these compounds on isolated guinea-pig atria are in good correlation with the data obtained from binding studies on guinea-pig ventricular homogenate as well as that obtained from comparative experiments on Na+,K+-ATPase activity inhibition. PMID- 2990492 TI - Antiproliferative properties of phorbol ester tumour promoters. PMID- 2990494 TI - A novel biologically active seleno-organic compound--V. Inhibition by ebselen (PZ 51) of rat peritoneal neutrophil lipoxygenase. AB - Suspensions of rat peritoneal PMNLs elicited with glycogen were stimulated by calcium and an ionophore to produce leukotrienes from endogenous arachidonic acid. We investigated the effect of the non-toxic, anti-inflammatory seleno organic compound, ebselen (PZ 51). When ethanolic extracts of the medium of stimulated cells were analysed by HPLC, a dose-dependent inhibition by ebselen of LTB4 formation with a concomitant decrease of 5-HETE production was found. Half maximum inhibition was observed at 20 mumoles/l ebselen. Similar findings were obtained after analysis of chloroform extracts of both cells and medium using a different HPLC system. Under these conditions, enhanced 5-HETE formation was associated with reduced production of LTB4 and other di-HETE isomers, when purified glutathione peroxidase + GSH were present. We conclude that the reported GSH peroxidase-like activity of ebselen, catalysing the reduction of 5-HPETE to 5 HETE, can not account for our findings. Therefore, the lipoxygenase reaction itself apparently represents the site of inhibition of LTB4 formation by ebselen. PMID- 2990493 TI - Effects of ethanol administration on rat liver plasma membrane-bound enzymes. AB - Changes in the properties of rat liver plasma membranes were examined in studies designed to differentiate between direct and metabolic effects of acute and chronic ethanol ingestion. One hour after a single dose of ethanol (3 g/kg body weight) there were increases in Na+,K+-ATPase (32%) and 5'-nucleotidase (36%), and hepatic concentrations of ethanol and acetaldehyde were approximately 23 mM and 50 microM, respectively. Na+,K+-ATPase and 5'-nucleotidase activities in liver plasma membranes from control rats were not significantly changed by in vitro addition of 30 microM acetaldehyde or 50 mM ethanol. Increases in Na+,K+ ATPase (approximately 20%) and 5'-nucleotidase (approximately 30%) were also observed in liver plasma membranes isolated from rats 16 hr after feeding ethanol or sucrose supplements for 17 days. The intake of calories from dietary protein and lipid was decreased by about 25% in both the ethanol and sucrose-fed animals. Na+,K+-ATPase activities in liver plasma membranes isolated from control rats were inhibited (approximately 20%) by 100 mM ethanol in vitro, whereas no inhibition was observed using membrane preparations from rats fed ethanol or sucrose supplements. Our results show that changes in liver plasma membrane enzyme activities associated with a single dose of ethanol are not a direct effect correlated with blood, hepatic or plasma membrane concentrations of ethanol or acetaldehyde. Chronic ingestion of ethanol or sucrose supplements had similar effects on liver plasma membrane enzyme characteristics and parallel changes in nutrient intake may be a more feasible explanation of these results than any analogous direct effects of the two compounds. PMID- 2990495 TI - Comparison of postnatal changes in alpha 1-adrenoceptor binding and adrenergic stimulation of phosphoinositide hydrolysis in rat cerebral cortex. AB - Adrenergic stimulation of phosphoinositide hydrolysis is mediated by the alpha 1 adrenoceptor subtype in many tissues including the brain. We have investigated the coupling of alpha 1-adrenoceptors to phosphoinositide hydrolysis during ontogeny. Alpha 1-adrenoceptor number and affinity were measured using [3H]prazosin binding in crude membranes of cerebral cortex and compared to the ability of the adrenergic agonists norepinephrine (NE) and phenylephrine (PE) to stimulate the formation of [3H] inositol phosphates from [3H]myo-inositol in brain slices at various ages. The greatest changes in the developmental expression of both the Bmax for [3H]prazosin binding and maximal (10(-4)M) NE- or PE-stimulated [3H]inositol phosphates were observed during the period of 7-21 days of age. No changes in the KD for [3H]prazosin were observed. However, at 14 days of age the EC50 for NE but not PE stimulation of [3H]inositol phosphates was slightly but significantly lower than at later ages. To quantitatively compare these two parameters during ontogeny, data were expressed as a percentage of the adult (greater than 65 day) value. At early ages (7 and 14 days) but not at later ages (21 and 37 days) the percent expression of [3H]prazosin binding sites was significantly greater than the maximal NE-stimulated [3H]inositol phosphates. This suggests that early in neonatal development alpha 1 adrenoceptors in brain are not tightly coupled to phosphoinositide hydrolysis. PMID- 2990496 TI - Comparison of aryl hydrocarbon hydroxylase and acetanilide 4-hydroxylase induction by polycyclic aromatic compounds in human and mouse cell lines. AB - The human MCF-7 and the mouse Hepa-1 cell culture lines were compared for aryl hydrocarbon hydroxylase and acetanilide 4-hydroxylase inducibility by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) and benzo[a]anthracene (BA) and TCDD- and BA specific binding in the cytosol and nucleus. The effective concentration of BA in the growth medium required to induce either enzyme to 50% of its maximally inducible activity (EC50) was the same (5-11 microM) in both MCF-7 and Hepa-1 cells. On the other hand, the EC50 for TCDD in MCF-7 cells (5-25 nM) was more than 40-fold greater than that in Hepa-1 cells (0.4 to 0.6 nM). P1-450- and P3 450-specific mouse cDNA probes were used to quantitate mRNA induction in the Hepa 1 cell line. P1-450 mRNA was induced markedly by TCDD and benzo[a] anthracene, whereas P3-450 mRNA was induced negligibly. A P1-450-specific human cDNA probe was used to quantitate P1-450 mRNA induction in the MCF-7 cell line. Aryl hydrocarbon hydroxylase inducibility by TCDD or BA always paralleled P1-450 mRNA inducibility in either the mouse or human line. Although the cytosolic Ah receptor in Hepa-1 cells was easily detected by sucrose density gradient centrifugation, gel permeation chromatography, and anion-exchange high performance liquid chromatography, the cytosolic receptor cannot be detected in MCF-7 cells. Following in vivo exposure of cultures to radiolabeled TCDD, the intranuclear concentration of inducer-receptor complex was at least fifty times greater in Hepa-1 than MCF-7 cultures. The complete lack of measurable cytosolic receptor and almost totally absent inducer-receptor complex in the nucleus of MCF 7 cells was, therefore, out of proportion to its capacity for aryl hydrocarbon hydroxylase and acetanilide 4-hydroxylase inducibility. This MCF-7 line should provide an interesting model for a better understanding of the mechanisms of drug metabolizing enzyme induction by polycyclic aromatic compounds, including the Ah receptor-mediated mechanism. PMID- 2990497 TI - A possible mechanism of action of thalidomide on rheumatoid arthritis. PMID- 2990498 TI - Growth factors, oncogenes and transformation. Part I: Growth factors and cell cycle control. PMID- 2990499 TI - [Contribution of monoclonal antibody D 47 in the study of sweat gland pathology]. AB - D 47 is a monoclonal antibody (IgG2) reacting with a surface antigen of cortical thymocytes. On normal human skin, D 47 was found to react with a cytoplasmic antigen of the cells of the secretory portion of eccrine sweat glands (ESG). No reactivity with D 47 is detected on the excretory part of ESG, on apocrine glands and all other cutaneous structures. In this work we studied through an indirect immunofluorescence method on frozen skin sections the reactivity pattern of D 47 on a group of epithelial skin tumours of certain or alleged glandular differentiation. These consisted of: eccrine spiradenoma (ES, 1 case); chondroid syringoma (CS, 1 case); syringomas of the eyelid (1 case); clear-cell hidradenocarcinoma (1 case); eccrine porocarcinoma (1 case); naevus sebaceus of Jadassohn (2 cases, one of which associated to a syringocystadenoma papilliferum); clear-cell acanthoma (1 case); extramammary Paget's disease (3 cases); basal-cell epithelioma (2 cases). D 47 yielded in a strong labelling of a significant proportion of cells in the cases of ES and CS, while on the remaining tumours, apart from normal ESG occasionally present in the peritumoral connective tissue, no reactivity was seen. From this study it becomes clear that D 47 represents an immunohistologic marker of eccrine-secretory differentiation and that it can be applied in the investigation of the differentiation and of the differential diagnosis of sweat-gland neoplasms. PMID- 2990500 TI - Effect of norepinephrine on inhibition of mouse brain (Na+ + K+)-stimulated, (Mg++)-dependent, and (Ca++)-dependent ATPase activities by ethanol. AB - Norepinephrine (0.1 mM) has been reported to "sensitize" (Na+ + K+)-ATPase activity of rat brain homogenates to inhibition by ethanol. The present study extends these investigations to the mouse and includes other ATPase activities. We measured the effects of norepinephrine on the sensitivity of ethanol-induced inhibition of (Na+ + K+)-stimulated (E.C. 3.6.1.3), (Mg++)-dependent (E.C. 3.6.1.4) and (Ca++)-dependent ATPase activities. Whole forebrains from C57BL/6J mice were homogenized and assayed in vitro for ATPase activity using standard conditions. Ethanol (0.125-2.0 M) caused a dose-dependent inhibition of all three ATPases. Norepinephrine (0.1 mM) had no appreciable effect on ethanol's inhibition of (Na+ + K+)-stimulated or (Ca++)-dependent ATPase activities, but slightly antagonized ethanol's effect on (Mg++)-ATPase. These results suggest that norepinephrine has little effect on the sensitivities of specific ATPases to ethanol inhibition in mouse brain. PMID- 2990501 TI - Dependence of ethanol-induced redox shift on hepatic oxygen tensions prevailing in vivo. AB - Since many metabolic derangements induced by ethanol have been linked to the redox shift, we studied the effects of oxygen in the range of tensions prevailing in vivo along the sinusoids on both the basal redox state and the shift induced by ethanol. The redox state of nicotinamide nucleotides was assessed by surface NADH fluorescence and that of cytochrome oxidase by transmittance dual-wavelength spectrophotometry in the hemoglobin-free, perfused rat liver. In the absence of ethanol, varying the oxygen tensions within the physiological range produced a redox gradient of both cytochrome oxidase and NAD+, with a more reduced state at tensions normally prevailing in perivenular zones. The degree of reduction of cytochrome oxidase at these physiological oxygen tensions was associated with no impairment in the ability of the liver to consume oxygen and to produce ATP, suggesting lack of cellular anoxia. 25 mM ethanol increased hepatic oxygen consumption, but had no direct effect on the state of reduction of cytochrome oxidase. The effects of ethanol and oxygen tensions on NADH fluorescence were additive, indicating that a greater redox shift should occur when ethanol is oxidized at oxygen tensions similar to those normally prevailing in prievenular zones than at those in periportal zones. This dependence of the ethanol-induced redox shift on oxygen tensions may contribute to the selective perivenular hepatotoxicity of alcohol. PMID- 2990502 TI - Chronic ethanol induces changes in opiate receptor function and in met-enkephalin release. AB - Ethanol induces supersensitivity of striatal delta-opiate receptor sites labelled by 3H-Etorphine. This effect may be ascribed to the diminished enkephalin release detected in striatal slices after chronic ethanol consumption. On the other hand, Kd values for 3H-Met-enkephalin and 3H-DHM (mu-opiate receptors) specific binding are enhanced. The different sensitivity of the two classes of opiate receptors to ethanol may be due to specific effects on enkephalinergic transmission. It has been hypothesized that the decrease of 3H-Met-enkephalin and 3H-DHM affinity for their receptors takes place because endogenous substances from ethanol metabolism (for example salsolinol) behave as mu opioid agonists. This hypothesis is confirmed by "in vitro" studies demonstrating that salsolinol displaces 3H-Met enkephalin and 3H-DHM but not 3H-DADLE binding. On the contrary, it seems that delta-receptors become supersensitive because of the decreased endogenous peptide release. PMID- 2990503 TI - Alcohol modulation of benzodiazepine receptors. AB - The effects of ethanol-feeding and in vitro ethanol addition on the binding of 3H diazepam and 3H-flunitrazepam in synaptosomal membrane preparations from rat cerebral cortex were investigated. Long-term ethanol-feeding (liquid-diet, 14 g/kg/day, pair-feeding, 35 days) significantly increased the diazepam dissociation constant (from 3.80 to 4.62 nM at 4 degrees C and from 29.3 to 35.2 nM at 37 degrees C). Similarly, the flunitrazepam dissociation constant was also increased (from 0.80 to 0.98 nM at 4 degrees C and from 6.52 to 8.90 nM at 37 degrees C). However, there was no significant difference in the number of receptors after ethanol feeding. Gamma-Aminobutyric acid (GABA) and pentobarbital increased the binding affinity by the same magnitude in control and ethanol-fed rats. Alcohol, in vitro (100 mM), did not have a significant effect on the binding parameters under most conditions. These findings suggest that long-term ethanol-feeding reduced the binding affinity of benzodiazepines without alteration of other properties of the receptors or their number. PMID- 2990504 TI - Binding characteristics of 3H-flunitrazepam and CL-218,872 in cerebellum and cortex of C57B1 mice made tolerant to and dependent on phenobarbital or ethanol. AB - The influence of chronic phenobarbital (PB) or chronic ethanol administration on binding characteristics of 3H-flunitrazepam (3H-FLU) in cerebellum and cortex of C57Bl mice was examined. Chronic PB treatment for six days decreased the number of binding sites (Bmax) for 3H-FLU, whereas no change in the affinity (KD) was found. Further kinetic analysis revealed that the overall decrease in Bmax was due to a reduced number of high affinity (Type 1) benzodiazepine (BDZ) binding sites in the cerebellum, but to a decreased number of low affinity (Type 2) BDZ binding sites in the cortex. Furthermore, a marked reduction in the pentobarbital produced enhancement of 3H-FLU binding was observed in the cerebellum of the PB treated animals. Following chronic ethanol administration for seven days, no change in the Bmax or in the KD could be demonstrated. However, in chronically ethanol-treated mice, the pentobarbital-induced stimulation of 3H-FLU binding was reduced in the cerebellum of mice 24 hours after discontinuation of the ethanol treatment. The significance of the present findings for the development of tolerance to and dependence on barbiturates and ethanol is discussed. PMID- 2990505 TI - Chronic ethanol exposure alters dopaminergic signal transduction processes. AB - A number of data suggest that the chronic ethanol treatment induces derangements of cell membrane structure leading to modifications of membrane related processes. In particular, alterations have been observed in the mechanisms of neurotransmitter recognition and in the coupling of the receptor with the effector system. Phosphorylation of specific proteins by cyclic AMP stimulated protein kinases represent the final step in the biological response in several distinct functional processes. Ethanol neurotoxic action therefore may affect neurotransmitter availability and release as well as receptors effector systems and protein phosphorylation. In this line, chronic ethanol treatment in rats decreases cyclic AMP dependent protein kinase activity in rat striatal membrane fractions. When lysine rich histone type III was used as exogenous substrate, cyclic AMP stimulated 32P incorporation was still decreased in the ethanol group. These data favor the hypothesis of a decreased capability of the enzyme to phosphorylate in response to cAMP. PMID- 2990506 TI - Stereospecific interaction between alcohol and opiates mediated by the mu high affinity binding receptor. AB - Experiment 1 revealed that preexposure to morphine blocked ethanol-induced CTA, and that conversely, preexposure to ethanol blocked morphine-induced CTA. In experiment 2, naloxazone injected 4.5 hr before the preexposure drug significantly reversed the interaction between morphine and ethanol. The results of Experiment 3 showed that preexposure to levorphanol blocked both morphine- and ethanol-induced CTAs while dextrorphan at the same dose did not affect the CTAs. These findings are discussed in terms of the involvement of the opiate receptors and their ligands in opiate-ethanol interaction. PMID- 2990508 TI - [A case of motor sensory neuropathy associated with mental retardation, conductive deafness and cerebellar signs]. PMID- 2990507 TI - Differential CNS sensitivity to PIA and theophylline in long-sleep and short sleep mice. AB - Long sleep (LS) and short sleep (SS) mice have a differential sensitivity to the behavioral actions of an adenosine agonist, R-phenylisopropyl-adenosine (PIA) that parallels their differential sensitivity to the soporific effects of ethanol. In addition to being more sensitive to the sedative effects of PIA, LS mice also show a greater excitatory response to an adenosine antagonist, theophylline (1,3-dimethylxanthine). The brain concentrations of both PIA and theophylline following drug administration do not differ in LS and SS mice, suggesting that the central nervous system of the LS mouse is more sensitive to both adenosine receptor agonists and antagonists. However, LS and SS mice made tolerant to ethanol did not show cross-tolerance to PIA. These results suggest that genetic selection for ethanol sensitivity has resulted in a parallel CNS sensitivity to purinergic drugs, but that acute alterations in sensitivity due to the development of ethanol tolerance do not involve changes in purinergic systems. PMID- 2990509 TI - [Effects of S-adenosyl-L-methionine on experimental cerebral ischemia]. AB - Effects of S-adenosyl-L-methionine (SAMe) on experimental cerebral ischemia were investigated using two different ischemic models. Cerebral energy metabolites (ATP, lactate, c-AMP) and brain water content were measured. It is reported that SAMe accelerates synthesis of phosphatidyl choline and increases erythrocyte membrane fluidity. Complete ischemia was produced by heart excision using wistar kyoto rats. SAMe (100 mg/kg, I.P.) was administered twice at one hour and immediately before inducing ischemia. The brain of rats were irradiated by microwave to stop the enzyme activity exactly 60 seconds after inducing ischemia and brain energy metabolites were measured. Recirculation model was produced by one hour recirculation following two hours ischemia induced by clipping of bilateral common carotid arteries using stroke-prone spontaneously hypertensive rats. SAMe (100 mg/kg, I.V.) was administered twice one hour after clipping and ten minutes after recirculation. The brain metabolites and water content were measured one hour after recirculation. In the complete ischemia, ATP and c-AMP levels were statistically high in the SAMe treated group compared to the untreated group (vehicle). But there was no statistical difference in lactate between the treated group and the untreated group. In the recirculation model, lactate elevation was suppressed in the SAMe treated group compared to the vehicle group with statistical difference, but there was no difference in ATP and c-AMP. Also, there was no difference in water content between the treated and the untreated group. SAMe protected energy failure in ischemia and accelerated recovery from ischemia. It is indicated that this agent is beneficial for treatment of cerebral ischemia in the acute stage. PMID- 2990510 TI - [Systemic blood pressure and respiration during plateau wave phenomena]. AB - The intracranial pressure, systemic blood pressure, respiratory pattern and heart rate were studied polygraphically in five patients with plateau waves in continuous intracranial pressure recording. Three brain-tumor patients and two benign intracranial hypertension patients were included. The intracranial pressure was recorded through an indwelling ventricular catheter attached to a pressure transducer. The systemic blood pressure was recorded through an intraarterial catheter placed in the femoral artery. The recordings were made from 18 to 40 hours and a total of 770 plateau waves were examined in this study. The systemic blood pressure showed little or no change in spite of a marked rise in intracranial pressure. The respiratory alterations were always noted during the plateau waves. This respiratory changes during the plateau waves were assigned into three patterns. The first was the suppressed respiratory pattern during the entire course of the plateau waves (Type I): the second was the pattern characterized by the initial suppression followed by the late hyperventilation (Type II); and the third was the hyperventilation pattern during the entire course of the plateau waves (Type III). The pattern of Type I was noticed in 65% of the observed plateau waves, the pattern of Type II in 23%, and the pattern of Type III in 12%. The heart rate decreased in most plateau waves, but there was little change in the heart rate in some plateau waves.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990511 TI - Non-invasive assessment of pulmonary blood supply after staged repair of pulmonary atresia. AB - Radionuclide studies were performed to determine pulmonary blood flow in six children who had undergone surgery for pulmonary atresia, ventricular septal defect, and hypoplastic pulmonary arteries with or without major aortopulmonary collateral arteries. Lung blood flow was assessed from both particle perfusion lung scans and the pulmonary and systemic phase of a radionuclide dynamic flow study. Five patients had perfusion defects identified on the particle perfusion lung scan. In three of these, abnormal areas were perfused only during the systemic phase of the flow study, a combination of findings that indicate the presence of perfusion by collateral arteries. In one patient no systemic perfusion was noted and in one an initial particle perfusion study indicated the presence of a lung segment perfused by a collateral artery. In this last patient the particle perfusion scan after total correction showed a reduction in the size of the lung perfusion defect and no evidence of lung perfusion during the systemic phase of the flow study. The particle perfusion lung scan in the sixth patient showed pronounced asymmetry in blood flow to the lungs with no segmental perfusion defect on the particle perfusion scan and no abnormalities on the systemic flow study. It is concluded that radionuclide lung perfusion and flow studies provide useful information on lung perfusion and merit further evaluation to define their role in the management of these patients. PMID- 2990512 TI - Exercise, cyclic AMP and malignant hyperpyrexia susceptibility. PMID- 2990513 TI - The irritable colon syndrome. PMID- 2990514 TI - Treatment of hepatocellular carcinoma with recombinant leucocyte interferon: a pilot study. PMID- 2990515 TI - Lectin-binding sites in Paget's disease. AB - The presence and distribution of lectin-binding sites on neoplastic cells of Paget's disease was studied using fluorescein isothiocyanate (FITC)-conjugated peanut agglutinin (PNA), and FITC-conjugated wheatgerm agglutinin (WGA), and compared with such lectin-binding sites on keratinocytes, and cells of eccrine glands, apocrine glands, and mammary glands. Neoplastic cells of both mammary and extramammary Paget's disease showed cytoplasmic staining with both lectins. There were however fewer stained cells in mammary Paget's disease than in extramammary Paget's disease. The cytoplasmic staining of lectin-binding sites in cells of apocrine glands was in sharp contrast to the cell-surface staining seen on keratinocytes, or cells of eccrine glands or mammary glands. These results indicate that the lectin-binding sites of neoplastic cells of Paget's disease more closely resemble those of cells of apocrine glands than of keratinocytes, cells of eccrine glands or cells of mammary glands. PMID- 2990516 TI - Leukotrienes B4, C4 and D4 stimulate DNA synthesis in cultured human epidermal keratinocytes. AB - Leukotrienes in psoriatic skin lesions are potent mediators of inflammation. We have studied the capacity of leukotrienes to stimulate the DNA synthesis of cultured human epidermal keratinocytes. At concentrations ranging from 10(-12) to 10(-8) M, LTB4 produced a 100% increase of DNA synthesis determined both as the incorporation of [3H] thymidine and as the labelling index. In comparison, LTB4 had no effect on the DNA synthesis of dermal fibroblast cultures. 5S,12S-LTB4 and 5S,12S-all-trans-LTB4 did not change the DNA synthesis of keratinocytes, but the effect of LTB4 was abolished in the presence of 5S,12S-all-trans HLTB4. Being less potent than LTB4 the peptidoleukotrienes (LTC4, LTD4) also stimulated keratinocyte DNA synthesis. The effect of the peptidoleukotrienes, but not of LTB4, was antagonized by FPL 55712. These results show that leukotrienes B4, C4 and D4 exert potent and stereospecific mitogenic effects on cultured human keratinocytes. The presence of these arachidonic acid metabolites in psoriatic skin lesions may be pertinent to both inflammation and aberrant epidermal growth in psoriasis. PMID- 2990517 TI - Generation of leukotrienes from normal epidermis and their demonstration in cutaneous disease. AB - In order to evaluate the significance of chemotactic leukotrienes in cutaneous disease, synthetic leukotriene B4 and its metabolites were examined during in vitro chemotaxis. Leukotriene B4, and less so 20-OH-leukotriene B4, were chemotactic for neutrophils and eosinophils, with a preferential attraction of eosinophils. The responsiveness of human monocytes towards leukotriene B4 was relatively low. Normal cells and cells from different patients varied in their quantitative response. Cells from patients with eczema and T-cell lymphoma tended to migrate less than those from patients with other inflammatory diseases. Leukotrienes are generated from several types of peripheral leukocytes. In order to examine whether resident cells of the skin can also produce these factors, isolated human and murine epidermal cells were examined for their ability to generate leukotriene B4 and leukotriene C4 in vitro. Arachidonic acid, and less so the ionophore A 23187, induced the generation of both types of factors, based on the finding in the bioassay, reverse-phase high-pressure liquid chromatography and radioimmunoassays. The same factors were demonstrated by either or all of these methods in skin biopsies, scales, blisters or suction blisters of patients with psoriasis, pitysiasis rubra pilaris, dyshidrosis, bullous pemphigoid, pressure urticaria, urticaria pigmentosa and drug reactions, but not in Sezary syndrome nor in callus and skin biopsies of normal controls. These findings underline the fact that the leukotrienes are potent inflammatory mediators in diverse skin diseases, but that they are not limited to any specific disease. Furthermore, a relationship between leukotrienes and tissue eosinophilia does not exist. PMID- 2990518 TI - The transition from acute to chronic inflammation. PMID- 2990519 TI - A simple method for studying chemotaxis, vascular permeability and histological modifications induced by mediators of inflammation in vivo in man. AB - A new method is described allowing the quantitative and kinetic analysis, in vivo in human skin, of the chemotaxis of inflammatory cells, of the modifications of vascular permeability and of the histological and cytological events induced by certain mediators of inflammation. In order to illustrate the usefulness of the method, the effects of chronic contact to human skin of two potent mediators of inflammation, LTB4 and Paf-acether, are described. PMID- 2990520 TI - Pharmacological actions of leukotrienes in the skin. PMID- 2990521 TI - The role of chemo-attractant lipoxygenase products in the pathogenesis of psoriasis. PMID- 2990522 TI - The iron-chelating agent picolinic acid enhances transferrin receptors expression in human erythroleukaemic cell lines. AB - Picolinic acid, a metal chelating molecule, was administered to human erythroleukaemic cell lines (K 562 and HEL) that were grown in serum-containing media. Picolinic acid inhibited both iron uptake and cell growth. Furthermore, picolinic acid was shown to markedly decrease the level of ferritin in the cells. In spite of the inhibition of cell growth, picolinic acid induced a marked increase in the transferrin-binding capacity of the cells. This phenomenon was due to a two-five-fold enhancement of the rate of transferrin receptor biosynthesis. Other iron-chelating compounds, capable of reducing the level of intracellular iron, also elicited a marked enhancement of the transferrin-binding capacity of the cells. However, the addition of iron, as ferric ammonium citrate, in the culture medium elicited a marked increase in the level of ferritin and a strong decrease in the transferrin-binding capacity of the cells. On the basis of these data we propose that a feed-back mechanism is involved in the regulation of transferrin receptors: when the cells accumulate iron they decrease the number of transferrin receptors in order to prevent further accumulation of iron; when no or low iron is available to the cells, the number of transferrin receptors markedly increases as a compensatory mechanism. PMID- 2990523 TI - Antibodies to human adult T cell leukaemia virus type I associated antigens in Swedish leukaemia patients and blood donors. AB - Antibodies to antigens associated with human T cell leukaemia virus type I (HTLV I) in Swedish adult leukaemia patients and blood donors were sought with a sensitive screening test using membrane antigen prepared from virus producing cells (MA-ELISA). Four persons (one ALL, one AML and two healthy blood donors) out of 483 persons tested reacted in the test. However, they were negative in the more specific anti-p19 and anti-whole virion ELISA tests. The prevalence of sera with definite anti-HTLV I activity seems to be very low in Sweden. The finding of four MA-ELISA positive persons needs further investigation. PMID- 2990524 TI - Compensation, radiographic changes, and survival in applicants for asbestosis compensation. AB - The survival of 354 claimants for compensation for pulmonary asbestosis among former workers of the Wittenoom crocidolite mine and mill in Western Australia has been examined. There were 118 deaths up to December 1982. The median time between start of work and claim for compensation was 17 years. The standardised mortality ratio (SMR) for deaths from all causes was 2.65 (p less than 0.0001). The SMR for pneumoconiosis was 177.2 (p less than 0.0001), bronchitis and emphysema 2.6 (p = 0.04), tuberculosis 44.6 (p less than 0.0001), respiratory cancer (including five deaths from malignant pleural mesothelioma) 6.4 (p less than 0.0001), gastrointestinal cancer 1.6 (p = 0.22), all other cancers 1.6 (p = 0.17), heart disease 1.4 (p = 0.07), and all other causes 2.18 (p = 0.004). Plain chest radiographs taken within two years of claiming compensation were found for 238 subjects and were categorised independently by two observers according to the International Labour Organisation criteria without knowledge of exposure or compensation details. Profusion of radiographic opacities, age at claiming compensation, work in the Wittenoom mill, and degree of disability awarded by the pneumoconiosis medical board were significant predictors of survival, but total estimated exposure to asbestos was not. Radiographic profusion and degree of disability were, however, predictable by total exposure. The median survival from claim for compensation was 17 years in subjects with ILO category 1 pneumoconiosis, 12 years in category 2, and three years in category 3. PMID- 2990525 TI - Occupational asthma in a steel coating plant. AB - An outbreak of occupational asthma, of unknown cause and extent, was detected in a steel coating plant. In 1979 a cross-sectional study which defined occupational asthma in terms of respiratory symptoms detected 21 people with suggestive symptoms among the 221 studied. They all worked in the coating shop, but the plastic coatings used at the plant contained many potential sensitising agents that might have caused the asthma. All 21 developed their symptoms after 1971, and it was found that in this year a supplier had modified a coating allowing, at the temperatures used in the process, toluene di-isocyanate to be liberated. Two of the symptomatic subjects were tested by inhalation of the isocyanate and showed asthmatic reactions and other subjects were found to have asthma related to periods spent at work by records of peak expiratory flow rate. Over half the 21 had a symptom free latent period after first exposure of three years or less, a pattern not seen in other subjects with respiratory symptoms. After the isocyanate had been removed from the process 17 of these subjects became asymptomatic or improved, a greater proportion than in other subjects with respiratory symptoms. PMID- 2990526 TI - Severe diffuse small airways abnormalities in long term chrysotile asbestos miners. AB - To determine the nature and extent of pathological changes in the small airways induced by asbestos, the pathological lesions of fibrosis and pigmentation of the membranous and respiratory bronchioles and alveolar ducts in lungs obtained from necropy from a group of 36 non-asbestotic long term chrysotile miners and 36 age, sex, and smoking matched controls who had no history of exposure to dust were compared. The airways were graded using a standard visual grading system. Appreciably greater airway fibrosis was found for all types of airway in the whole group of miners compared with the controls. Differences for pigmentation were generally similar but less pronounced. It is concluded that long term exposure to mineral dust as an asbestos miner produces severe diffuse pathological changes in the small airways; these findings may relate to the physiological observation of unusual degrees of airflow obstruction in this group of workers. PMID- 2990527 TI - Beauvaria bassiana keratitis. AB - The cornea of a 64-year-old white male underwent progressive thinning following removal of a foreign body and after treatment with topical antibiotics and corticosteroid. Initial attempts at laboratory identification of an infectious agent were negative. The process progressed to corneal perforation. After a penetrating keratoplaty, histopathological examination of host button tissue showed a fungus, identified as Beauvaria bassiana on culture. PMID- 2990528 TI - Displacement of DL-[3H]-2-amino-4-phosphonobutanoic acid ( [3H]APB) binding with methyl-substituted APB analogues and glutamate agonists. AB - The binding of the excitatory amino acid antagonist DL-2-amino-4 phosphonobutanoic acid (DL-APB) to rat brain synaptic plasma membranes was characterized. As determined by Scatchard analysis, the binding was saturable and homogeneous with a Kd = 6.0 microM and Bmax = 380 pmol/mg of protein. The binding was dependent on the presence of Ca2+ and Cl- ions and was diminished upon freezing. The association rate constant was 6.8 X 10(-3) microM-1 min-1, and the dissociation rate constant was 2.0 X 10(-2) min-1. The L isomers of APB, glutamate, and aspartate were more potent as displacers of APB binding than the D isomers. Previously determined inhibition data obtained for APB-sensitive inputs to hippocampal granule cells are compared to the present displacement data in an attempt to identify this binding protein as the recognition site of the receptor mediating the APB-induced inhibition of synaptic transmission. With the exception of kynurenic acid, all compounds examined in both systems were more potent as displacers of APB binding than as inhibitors of synaptic transmission. This difference in potency was most pronounced for agonists at dentate granule cells. L-Glutamate, D-glutamate, and L-glutamate tetrazole were between 140- and 7500 fold more potent as displacers of DL-APB binding than as inhibitors of synaptic transmission. D-2-Amino-5-phosphonopentanoic acid and alpha-methyl-APB were between 10- and 20-fold more potent as displacers of binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990529 TI - Dynamic 13C NMR investigations of substrate interaction and catalysis by cobalt(II) human carbonic anhydrase I. AB - Using 13C NMR spectroscopy, we have further investigated the binding of HCO3- in the active site of an artificial form of human carbonic anhydrase I in which the native zinc is replaced by Co(II). The Co(II) enzyme, unlike all other metal substituted derivatives, has functional properties closely similar to those of the native zinc enzyme. By measuring the spin-lattice relaxation rate and the line width for both the CO2 and HCO3- at two field strengths, we have determined both the paramagnetic effects that reflect substrate binding and the exchange effects due to catalysis at chemical equilibrium. The following are the results at 14 degrees C and pH 6.3 (1) HCO3- is bound in the active site of the catalytically competent enzyme with the 13C of the HCO3- located 3.22 +/- 0.02 A from the Co(II); (2) the apparent equilibrium dissociation constant for the bound HCO3- is 7.6 +/- 1.5 mM, determined by using the paramagnetic effects on the line widths, and 10 +/- 2 mM, determined by using the exchange effects; (3) the lifetime of HCO3- bound to the metal is (4.4 +/- 0.4) X 10(-5) s; (4) the overall catalyzed CO2 in equilibrium HCO3- exchange rate constant of the Co(II) enzyme is (9.6 +/- 0.4) X 10(3) s-1; (5) the electron spin relaxation time of the Co(II), determined by using paramagnetic effects on the bound HCO3-, is (1.1 +/- 0.1) X 10(-11) s. The data did not provide any direct information on the binding of CO2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990531 TI - Multinuclear magnetic resonance studies of metal ion binding sites of phosphoglucomutase. AB - Metal binding at the activating site of rabbit muscle phosphoglucomutase has been studied by 31P, 7Li, and 113Cd NMR spectroscopy. A 7Li NMR signal of the binary Li+ complex of the phosphoenzyme was not observed probably because of rapid transverse relaxation of the bound ion due to chemical exchange with free Li+. The phosphoenzyme-Li+-glucose 6-phosphate ternary complex is more stable, kinetically, and yields a well-resolved peak from bound Li+ at -0.24 ppm from LiCl with a line width of 5 Hz and a T1 relaxation time of 0.51 +/- 0.07 s at 78 MHz. When glucose 1-phosphate was bound, instead, the chemical shift of bound 7Li+ was -0.13 ppm; and in the Li+ complex of the dephosphoenzyme and glucose bisphosphate a partially broadened 7Li+ peak appeared at -0.08 ppm. Thus, the bound metal ion has a somewhat different environment in each of these three ternary complexes. The 113Cd NMR signal of the binary Cd2+ complex of the phosphoenzyme appears at 22 ppm relative to Cd(ClO4)2 with a line width of 20 Hz at 44.4 MHz. Binding of substrate and formation of the Cd2+ complex of the dephosphoenzyme and glucose bisphosphate broaden the 113Cd NMR signal to 70 Hz and shift it to 75 ppm. The 53 ppm downfield shift upon the addition of substrate along with 1H NMR data suggests that one oxygen ligand to Cd2+ in the binary complex is replaced by a nitrogen ligand at some intermediate point in the enzymic reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990530 TI - Mobility in the mitochondrial electron transport chain. AB - The role of lateral diffusion in mitochondrial electron transport has been investigated by measuring the diffusion coefficients for lipid, cytochrome c, and cytochrome oxidase in membranes of giant mitoplasts from cuprizone-fed mice using the technique of fluorescence redistribution after photobleaching (FRAP). The diffusion coefficient of the phospholipid analogue N-(7-nitro-2,1,3-benzoxadiazol 4-yl)phosphatidylethanolamine is dependent on the technique used to remove the outer mitochondrial membrane. A sonication technique yields mitoplasts with monophasic recovery of the lipid probe (D = 6 X 10(-9) cm2/s), while digitonin treated mitochondria show biphasic recoveries (D1 = 5 X 10(-9) cm2/s; D2 = 1 X 10(-9) cm2/s). Digitonin appears to incorporate into mitoplasts, giving rise to decreased lipid mobility concomitant with increased rates of electron transfer from succinate to oxygen, in a manner reminiscent of the effects of cholesterol incorporation [Schneider, H., Lemasters, J. J., Hochli, M., & Hackenbrock, C. R. (1980) J. Biol. Chem. 255, 3748-3756]. FRAP measurements on tetramethylrhodamine cytochrome c modified at lysine-39 and on a mixture of active morpholinorhodamine derivatives of cytochrome c gave diffusion coefficients of (3.5-7) X 10(-10) cm2/s depending on the assay medium. With morpholinorhodamine-labeled antibodies purified on a cytochrome oxidase affinity column, the diffusion coefficient for cytochrome oxidase was determined to be 1.5 X 10(-10) cm2/s. The results are discussed in terms of a dynamic aggregate model in which an equilibrium exists between freely diffusing and associated electron-transfer components. PMID- 2990532 TI - Novel multilayered lipid vesicles: comparison of physical characteristics of multilamellar liposomes and stable plurilamellar vesicles. AB - The preparation of a new kind of multilayered liposome, called a stable plurilamellar vesicle (SPLV), is described. Although SPLVs and classical multilamellar vesicles (MLVs) are made of the same materials and appear overtly similar in the electron microscope, the two types of vesicles differ as determined by stability, entrapment efficiency, electron spin resonance (ESR), NMR, X-ray diffraction, and biological effects. It is demonstrated that, contrary to what has been assumed, classical MLVs exclude solutes during their formation and, thus, are under a state of osmotic compression. By contrast, the SPLV process produces liposomes that are not compressed. The effects of osmotic compression are discussed. It is suggested that the state of osmotic stress is an important variable that distinguishes various types of liposomes and that has significant physical and biological consequences. PMID- 2990533 TI - Membrane bilayer balance and erythrocyte shape: a quantitative assessment. AB - When human erythrocytes are incubated with certain phospholipids, the cells become spiculate echinocytes, resembling red cells subjected to metabolic starvation or Ca2+ loading. The present study examines (1) the mode of binding of saturated phosphatidylcholines and egg lysophosphatidylcholine to erythrocytes and (2) the quantitative relationship between phospholipid incorporation and red cell shape. We find that the phospholipids studied become intercalated into erythrocyte membranes, not simply adsorbed to the cell surface. Spin-labeling and radiolabeling data show that the incorporation of (4 +/- 1) X 10(6) molecules of exogenous phosphatidylcholine per cell converts discocytes to stage 3 echinocytes with about 35 conical spicules. This amount of lipid incorporation is estimated to expand the red cell membrane outer monolayer by 1.7% +/- 0.6%. Calculations of the inner and outer monolayer surface areas of model discocytes and stage 3 echinocytes yield an estimated difference of 0.7% +/- 0.2%. PMID- 2990534 TI - Modulation of (Na+,K+)-ATPase activity by the lipid bilayer examined with dansylated phosphatidylserine. AB - The fluorescent probe 8-(dimethylamino)naphthalene-1-sulfonylphosphatidylserine (Dns-PS) was incorporated into purified lamb kidney Na+- and K+-stimulated adenosinetriphosphatase (EC 3.6.1.3) [(Na+,K+)-ATPase] by using a purified phospholipid exchange protein. Phospholipase C was used to reduce phospholipid content. Up to 40% of the phospholipid could be hydrolyzed with only 10% inhibition of the (Na+,K+)-ATPase, but when 67% of the phospholipid was hydrolyzed, the enzyme was inhibited 53%. To examine the effect of protein on the phospholipid bilayer, the fluorescent parameters of the probe incorporated into the enzyme preparation were contrasted with the same parameters for the probe incorporated into the total lipid extract of the preparation. The polarization of fluorescence of the probe in the lipid extract was 0.118 while in the enzyme preparation it was 0.218. This reflected a decrease in fluidity of the glycerol region of the phospholipid bilayer which was mediated by the protein. This effect increased as the phospholipid content of the (Na+,K+)-ATPase preparation was reduced so that with maximal phospholipid reduction the polarization of fluorescence was 0.262. The protein caused a decrease in the transition temperature from gel to fluid states of the bilayer detected by polarization of the probe. The midpoint temperature transition of the enzyme preparation decreased from 33 degrees C when all phospholipids were present to 20 degrees C when 67% of the phospholipids were hydrolyzed. This decrease was not observed for the lipid extract of these samples. A direct correlation between the (Na+,K+) ATPase specific activity and the polarization of fluorescence of Dns-PS was found. The reduction in phospholipid content did not affect the steady-state level of phosphorylation of the enzyme by ATP but did affect the rate of dephosphorylation which would require conformational changes of the enzymes. The data showed that the fluidity of the phospholipid bilayer can modulate the activity of the (Na+,K+)-ATPase. PMID- 2990535 TI - Decreased lipid order induced by microsomal cytochrome P-450 and NADPH-cytochrome P-450 reductase in model membranes: fluorescence and electron spin resonance studies. AB - Cytochrome P-450 and NADPH-cytochrome P-450 reductase were reconstituted in unilamellar lipid vesicles prepared by the cholate dialysis technique from pure dimyristoylphosphatidylcholine (DMPC), pure dipalmitoylphosphatidylcholine (DPPC), pure dioleoylphosphatidylcholine (DOPC), and phosphatidylcholine/phosphatidylethanolamine/phosphatidylserine (PC/PE/PS) (10:5:1). As probes for the vesicles' hydrocarbon region, 1,6-diphenyl-1,3,5 hexatriene (DPH) and spin-labeled PC were used. The steady-state and time resolved fluorescence parameters of DPH were determined as a function of temperature and composition of liposomes. Incorporation of either protein alone or together increased the steady-state fluorescence anisotropy (rs) of DPH in DOPC and PC/PE/PS (10:5:1) liposomes. In DMPC and DPPC vesicles, the proteins decreased rs significantly below the transition temperature (Tc) of the gel to liquid-crystalline phase transition. Time-resolved fluorescence measurements of DPH performed in reconstituted PC/PE/PS and DMPC proteoliposomes showed that the proteins disorder the bilayer both in the gel and in the liquid-crystalline phase. Little disordering by the proteins was observed by a spin-label located near the mid-zone of the bilayer 1-palmitoyl-2-(5-doxylstearoyl)-3-sn phosphatidylcholine (8-doxyl-PC), whereas pronounced disordering was detected by 1-palmitoyl-2-(8-doxylpalmitoyl)-3-sn-phosphatidylcholine (5-doxyl-PC), which probes the lipid zone closer to the polar part of the membrane. Fluorescence lifetime measurements of DPH indicate an average distance of greater than or equal to 60 A between the heme of cytochrome P-450 and DPH. PMID- 2990536 TI - Ubiquitin-dependent proteolysis of native and alkylated bovine serum albumin: effects of protein structure and ATP concentration on selectivity. AB - The susceptibility of bovine serum albumin to degradation by the ubiquitin dependent system of proteolysis depends on the severity of the iodination conditions [Wilkinson, K.D., & Audhya, T.K. (1981) J. Biol. Chem. 256, 9235 9241]. To evaluate if other modifications of the protein changed its susceptibility to degradation, chemically modified derivatives of bovine serum albumin have been synthesized, characterized, and tested as substrates for the ubiquitin-dependent system. Serum albumin was reduced or reduced and alkylated with iodoacetic acid or iodoacetamide. Only the alkylated derivatives exhibit saturation kinetics. Both alkylated proteins competitively inhibit the degradation of the other. These substrates are useful for assay of the intact proteolysis system in crude extracts and in assays for other substrates using competitive alternate substrate inhibition. The physical properties of these proteins suggest that charge, denaturation, or aggregation is not correlated with the degradation rate of these proteins by this system. However, the selectivity of the ubiquitin-dependent proteolysis depends strongly on the ATP concentration. At saturating substrate concentrations, both alkylated substrates are degraded equally. At low ATP concentrations, there is a 2.4-fold difference in the degradation rates of the alkylated proteins. The results presented here indicate that the ubiquitin-dependent protein degradation system is selective and responsive to ATP concentrations and that not all abnormal proteins are equally preferred substrates. Thus, the system may be more selective than previously thought. PMID- 2990537 TI - Effect of buffer on kinetics of proton equilibration with a protonable group. AB - The laser-induced proton pulse generates a massive, brief, proton pulse capable of perturbing biochemical equilibria. The time resolution of the monitoring system can follow the diffusion-controlled protonation of specific sites on macromolecular bodies [Gutman, M. (1984) Methods Biochem. Anal. 30, 1-103]. In order to apply this method in enzymology, one must first evaluate how the buffer capacity of biochemical systems (substrates and proteins) will affect the observed dynamics. Unlike equilibrium measurements, where buffer is an inert component, in kinetic studies buffer modulates the observed dynamics. In this paper we analyze the effect of buffer on the dynamics of protonation in a model system. We describe the experimental technique and introduce the mathematical formalism that determines the various rate constants involved in the reaction. The analysis of the experiments indicates that in buffered solution proton flux is carried by two mechanisms: (A) proton dissociation followed by free proton diffusion; (B) collisional proton transfer between small diffusing solutes. We demonstrate how to evaluate the contribution of each pathway to the overall proton flux. PMID- 2990538 TI - Kinetic analysis of protonation of a specific site on a buffered surface of a macromolecular body. AB - The kinetics of protonation of a specific site on a macromolecular structure (micelle) in buffered solution was studied with the purpose of evaluating the effect of buffer on the observed dynamics. The experimental system consisted of the following elements: Brij 58 micelles serving as homogeneous uncharged macromolecular bodies, bromocresol green, a well-adsorbed proton detector, and 2 naphthol-3,6-disulfonate as a proton emitter in the bulk. Imidazole was the mobile buffer while neutral red, which has a high affinity for the micellar surface, served as the immobile buffer. An intensive laser pulse ejects a proton from the proton emitter, and the subsequent proton-transfer reactions are measured by fast spectrophotometric methods. The dynamics of proton pulse in buffered solution are characterized by a very rapid trapping of the discharged protons by the abundant buffer molecules. This event has a major effect on the kinetic regime of the reaction. During the first 200 ns the proton flux is rate limited by free-proton diffusion. After this period, when the free-proton concentration decayed to the equilibrium level, the relaxation of the system is carried out by the diffusion of buffer. Thus in the buffered biochemical system, at neutral pH, most of proton flux between active sites and bulk is carried out by buffer molecules--not by diffusion of free protons. Surface groups on a high molecular weight body exchange protons among them at a very fast rate. This reaction has a major role on proton transfer from a specific site to the bulk.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990539 TI - Active and resting states of the O2-evolving complex of photosystem II. AB - During dark adaptation, a change in the O2-evolving complex (OEC) of spinach photosystem II (PSII) occurs that affects both the structure of the Mn site and the chemical properties of the OEC, as determined from low-temperature electron paramagnetic resonance (EPR) spectroscopy and O2 measurements. The S2-state multiline EPR signal, arising from a Mn-containing species in the OEC, exhibits different properties in long-term (4 h at 0 degrees C) and short-term (6 min at 0 degree C) dark-adapted PSII membranes or thylakoids. The optimal temperature for producing this EPR signal in long-term dark-adapted samples is 200 K compared to 170 K for short-term dark-adapted samples. However, in short-term dark-adapted samples, illumination at 170 K produces an EPR signal with a different hyperfine structure and a wider field range than does illumination at 160 K or below. In contrast, the line shape of the S2-state EPR signal produced in long-term dark adapted samples is independent of the illumination temperature. The EPR-detected change in the Mn site of the OEC that occurs during dark adaptation is correlated with a change in O2 consumption activity of PSII or thylakoid membranes. PSII membranes and thylakoid membranes slowly consume O2 following illumination, but only when a functional OEC and excess reductant are present. We assign this slow consumption of O2 to a catalytic reduction of O2 by the OEC in the dark. The rate of O2 consumption decreases during dark adaptation; long-term dark-adapted PSII or thylakoid membranes do not consume O2 despite the presence of excess reductant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990540 TI - Measurement of DNA-protein equilibria using gel chromatography: application to the HinfI restriction endonuclease. AB - A method is described for measuring equilibrium constants of DNA-protein interactions using gel chromatography. This technique has been used to study the sequence-specific interaction of the HinfI restriction endonuclease with DNA. HinfI has a monomeric molecular weight of 31000 and exists as a dimer in its active form. The protein binds to supercoiled DNA molecules containing its recognition site with an apparent free energy of -13.9 kcal/mol of sites. This interaction is highly salt sensitive and causes a release of 3.4 ion pairs. The affinity of the nuclease for its recognition site is largely independent of both pH (6.5-8.5) and temperature (7-35 degrees C) and was not affected by variations in the degenerate middle position of the site. Linear DNA fragments containing the HinfI recognition site were bound as tightly as supercoiled molecules. Binding to nonspecific DNA sites or to methylated DNA sites was approximately 6 orders of magnitude weaker. In general, enzyme activity and binding affinity paralleled each other. PMID- 2990541 TI - Fluoro ketone inhibitors of hydrolytic enzymes. AB - The use of fluoro ketones as inhibitors of hydrolytic enzymes has been investigated. The acetylcholine analogues 6,6-dimethyl-1,1,1-trifluoro-2 heptanone and 3,3-difluoro-6,6-dimethyl-2-heptanone are inhibitors of acetylcholinesterase with Ki values of 16 X 10(-9) M and 1.6 X 10(-9) M, respectively. These fluoro ketones are 10(4)-10(5) times better as inhibitors than the corresponding methyl ketone. Since nucleophiles readily add to fluoro ketones, it is likely that these compounds inhibit acetylcholinesterase by formation of a stable hemiketal with the active-site serine residue. Fluoro ketone substrate analogues are also inhibitors of zinc metallo- and aspartylproteases. 2-Benzyl-4-oxo-5,5,5-trifluoropentanoic acid is an inhibitor of carboxypeptidase A (Ki = 2 X 10(-7) M). Trifluoromethyl ketone dipeptide analogues are good inhibitors of angiotensin converting enzyme. An analogue of pepstatin that contains a difluorostatone residue in place of statine has been prepared and found to be an extremely potent inhibitor of pepsin (Ki = 6 X 10( 11) M). The hydrated ketones are probably the inhibitory species since they are structural mimics of the tetrahedral intermediate that forms during the hydrolysis of peptide substrates. PMID- 2990542 TI - Evidence for active intermediates during the reconstitution of yeast phosphoglycerate mutase. AB - The reconstitution of the tetrameric phosphoglycerate mutase from bakers' yeast after denaturation in guanidine hydrochloride has been studied. When assays are performed in the presence of trypsin, it is found that reactivation parallels the regain of tetrameric structure. However, in the absence of trypsin, the regain of activity is more rapid, suggesting that monomeric and dimeric intermediates possess partial activity (35% of the value of native enzyme) which is sensitive to trypsin. When reconstitution is studied in the presence of substrates, it is again found that monomeric and dimeric intermediates possess 35% activity. Under these latter conditions, the activity of the monomer but not of the dimer is sensitive to trypsin. PMID- 2990543 TI - Inhibition of soybean lipoxygenase 1 by N-alkylhydroxylamines. AB - Micromolar concentrations of N-octylhydroxylamine dramatically increase the induction period in the conversion of linoleic acid to 13(S)-hydroperoxy-cis 9,trans-11-octadecadienoic acid (13-HPOD) catalyzed by soybean lipoxygenase 1. The induction period produced by N-octylhydroxylamine is abolished by 13-HPOD but not by the corresponding hydroxy acid. Addition of a catalytic amount of lipoxygenase to a mixture of 13-HPOD and N-octylhydroxylamine results in consumption of approximately 1 mumol of 13-HPOD/mumol of N-octylhydroxylamine present. These results can be explained by a model in which 13-HPOD oxidizes the enzyme from an inactive ferrous form to an active ferric form, as proposed by previous workers, and N-octylhydroxylamine reduces the enzyme back to the ferrous form. Consistent with this model, the ESR signal at g = 6.1 characteristic of ferric lipoxygenase is rapidly abolished by N-octylhydroxylamine and can be regenerated by 13-HPOD. These results provide additional support for earlier proposals that ferric lipoxygenase is the catalytically active form and also establish a novel method of inhibiting enzymes in this class. The octyl group of N-octylhydroxylamine appears to contribute to binding near the iron, since hydroxylamine and N-methylhydroxylamine do not extend the induction period. In the n-RNHOH series, activity passes through an optimum at R = decyl. PMID- 2990544 TI - Differential effects of oxidizing agents on human plasma alpha 1-proteinase inhibitor and human neutrophil myeloperoxidase. AB - Human alpha 1-proteinase inhibitor is easily susceptible to inactivation because of the presence of a methionyl residue at its reactive site. Thus, oxidizing species derived from the myeloperoxidase system (enzyme, H2O2, and C1-), as well as hypochlorous acid, can inactivate this inhibitor, although H2O2 alone has no effect. Butylated hydroxytoluene, a radical scavenger, partially protects alpha 1 proteinase inhibitor from the myeloperoxidase system and completely protects it from hypochlorous acid. Each oxidant also reacts differently with the inhibitor, in that the myeloperoxidase system and hypochlorous acid can each oxidize as many as six methionyl residues, but hypochlorous acid can also oxidize a single tyrosine residue. Myeloperoxidase can be inactivated by hypochlorous acid, by autoxidation in the presence of H2O2 and C1-, as well as by H2O2 alone. Butylated hydroxytoluene completely protects this enzyme from hypochlorous acid inactivation, does not affect the action of H2O2, and enhances autoinactivation. As many as six methionyl residues and two tyrosine residues could be oxidized during autoxidation and six methionine residues by H2O2 alone. Eight methionine residues and one tyrosine residue could be oxidized by hypochlorous acid. The tyrosine residue in myeloperoxidase was oxidized only at a relatively high concentration (600 microM) of hypochlorous acid at which point the enzyme simultaneously and completely lost its enzymatic activity. Loss of activity of myeloperoxidase could also be correlated with the loss of the heme groups present in the enzyme when a relatively high concentration of hypochlorous acid (600 microM) was used and also during autoxidation. It appears that once there is sufficient oxidant to modify one of the tyrosine residues, the heme group itself becomes susceptible. PMID- 2990545 TI - Synthetic tools for adrenocorticotropin receptor identification. AB - Biotinylated photoaffinity derivatives of adrenocorticotropin (ACTH) are potentially useful tools for the identification of ACTH receptors. The hormone can be attached covalently to its receptor by photoactivation, and the presence of biotin in the molecule facilitates isolation of the solubilized hormone receptor complex on columns of immobilized succinoylavidin (Suc-avidin). Six photoprobes of ACTH1-24 have been prepared by reacting ACTH1-24, [25 biocytin]ACTH1-25 amide, and [25-dethiobiocytin]ACTH1-25 amide with either 4- or 5-azido-2-nitrophenylsulfenyl (4-NAPS and 5-NAPS, respectively) chlorides in acetic acid. The homogeneity of the photoprobes was carefully monitored by thin layer chromatography and amino acid analyses of acid hydrolysates. The presence of underivatized starting material in the photoprobes was critically scrutinized by high-pressure liquid chromatography and was estimated to be less than 0.5%. Both the 4- and 5-NAPS derivatives stimulated maximal steroidogenesis (as compared with ACTH1-24) in calf adrenal cortical cells. However, the potencies of the two isomers differed significantly. The ED50 for steroidogenesis with 5-NAPS ACTH1-24 was 100-fold greater than the standard (ACTH1-24) while that for 4-NAPS ACTH1-24 was only approximately 7 times greater. Although 4-NAPS-ACTH1-24 was capable of stimulating maximal adenosine cyclic 3',5'-phosphate (cAMP) production, the 5-NAPS derivative was usually not. The level of stimulation with the 5-NAPS derivative varied considerably from cell preparation to cell preparation. ACTH1-24-induced cAMP production was inhibited by 5-NAPS-ACTH1-24 or 5-NAPS-[25-dethiobiocytin]ACTH1-25 amide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990546 TI - Design, synthesis, and characterization of a model peptide having potent calcitonin-like biological activity: implications for calcitonin structure/activity. AB - A calcitonin analogue, MCT-II, having the potential to form an amphiphilic alpha helix from residue 8 to residue 22 with a continuous surface of aliphatic leucine side chains on the hydrophobic face of the helix has been synthesized, and its physical and biological properties have been characterized. Properties exhibited by this peptide, including self-association in the micromolar concentration range with a concomitant increase in the percentage of alpha-helical structure, formation of stable monolayers at the air-water interface, and adsorption to the surface of egg lecithin single-bilayer vesicles, demonstrate that MCT-II can readily form an amphiphilic alpha-helical structure. Though MCT-II has minimal sequence homology to any particular natural analogue from residue 8 to residue 22, it has biological activity similar to that of salmon calcitonin I for receptor binding in brain and kidney membranes, for activation of adenylate cyclase, and in hypocalcemic potency in vivo. The amphiphilic alpha-helical structure of MCT-II, therefore, is important for binding to calcitonin receptors. It is also apparent that a hydrophilic residue commonly occurring on the hydrophobic face (position 15) in the natural calcitonins is not required for high biological activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990547 TI - Carbon monoxide binding to Rhodospirillum molischianum ferrocytochrome c'. AB - Reversible carbon monoxide binding has been used to examine the structural and functional properties of reduced Rhodospirillum molischianum cytochrome c'. The symmetrical dimer is found to bind CO in a noncooperative manner, indicating that the heme sites function independently and with identical carbon monoxide affinity. The enthalpy change of binding CO (aqueous) to R. molischianum ferrocytochrome c' is determined to be -11 kcal/mol of CO, which is comparable to the heat of CO binding to other heme proteins. A Bohr effect is observed (0.31 +/ 0.04 proton released per mole of CO bound at pH 8), and a basic group is involved which changes its pK from 8.3 to 7.8 upon ligation. The histidine axial ligand to the heme iron is suggested to be the source of the Bohr effect. Increased CO affinities were observed at high pH or at neutral pH in the presence of phosphate. These solvent-induced changes in CO affinity do not appear to be caused by changes in quaternary structure but rather are more likely brought about by localized changes in the vicinity of the solvent-exposed heme face. PMID- 2990548 TI - Substrate-induced modifications of the intrinsic fluorescence of the isolated adenine nucleotide carrier protein: demonstration of distinct conformational states. AB - The effects of ATP or ADP and the specific inhibitors carboxyatractyloside (CATR) and bongkrekic acid (BA) on the conformation of the isolated adenine nucleotide (AdN) carrier protein were studied by fluorescence spectroscopy. The addition of ATP to the AdN carrier resulted in a rapid fluorescence increase of the tryptophanyl residue(s) at 355 nm, which leveled up in less than 1 s at 22 degrees C. Among the natural nucleotides, only ATP and ADP were effective. At 10 degrees C or below, the kinetics of the fluorescence increase induced by ATP were biphasic, consisting of a rapid phase of less than 1 s, followed by a slower phase that lasted for a few seconds and had virtually the same amplitude as the rapid one. Both phases were abolished when CATR was added prior to ATP or fully reversed when CATR was added after the fluorescence response to ATP had been elicited. The number of CATR binding sites present on the carrier protein was determined by CATR specific inhibition of the ATP-induced increase in intrinsic fluorescence. The calculated number of CATR sites was equal to that found by another method based on the use of the same preparation of AdN carrier loaded with fluorescent nucleotide naphthoyl-ATP and on the CATR-induced release of the bound naphthoyl-ATP, demonstrating the reliability of the intrinsic fluorescence assay. Addition of BA prior to or together with ATP nearly doubled the amplitude of the ATP-induced fluorescence signal. At 10 degrees C or below, the fluorescence response to ATP in the presence of BA could also be decomposed into rapid and slow phases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990549 TI - Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines. AB - Epstein-Barr (EB) virus induces a new pyrimidine deoxynucleoside kinase [thymidine kinase (dTk)] activity in Raji B lymphocyte cells after superinfection. This dTk activity is also present in small amounts in the HR-1 virus-producer cell line and in larger amounts in the B95-8 virus-producer line. The dTk activity induced by EB virus coelutes from DEAE-cellulose columns with deoxycytidine kinase (dCk) activity and elutes as a broad peak well separated from the large peaks of cellular dTk and dCk activities. This EB virus-induced pyrimidine deoxynucleoside kinase activity from HR-1 cells differs from cellular kinases in most basic biochemical properties but shares certain properties with the herpes simplex virus dTk. PMID- 2990550 TI - Nucleotide sequence of the gene for the gamma chain of human fibrinogen. AB - A human genomic DNA library was screened for the gene coding for the gamma chain of fibrinogen by using a human cDNA for the gamma chain as a hybridization probe. The gene was identified in three overlapping recombinant lambda bacteriophage, and its sequence, including the immediate 5' and 3' flanking regions, was determined. The DNA sequence analysis revealed the presence of 10 exons coding for 411 amino acids present in the mature protein and a signal sequence of 26 amino acids. Two 30 base pair (bp) direct repeats of 93% identity were found 468 bp upstream from the transcription initiation site. The DNA sequence of the gene for the gamma chain of human fibrinogen showed considerable sequence homology with a partial sequence reported for the gene for the gamma chain of rat fibrinogen. PMID- 2990552 TI - Complexity in the redox titration of the dihaem cytochrome c4. AB - Redox titration of the dihaem, two domain cytochromes c4 from Pseudomonas aeruginosa, Pseudomonas stutzeri and Azotobacter vinelandii showed complex behaviour indicative of the presence of two redox components. In the case of the P. stutzeri cytochrome c4, two spectroscopically distinct components were present during the redox titration. In contrast, cytochrome c-554(548) from a halophilic Paracoccus species is a stable dimer of a monohaem cytochrome which shows close homology to cytochrome c4, but does not show complexity in its redox titration. The presence of chemically distinct haem environments or anti-cooperative interactions between identical haem groups are two possible explanations for the redox complexity of cytochrome c4. The simple redox titration of cytochrome c 554(548) shows that haems situated relatively close together need not interact, but direct cleavage, separation and study of the domains will be necessary to decide whether they do or do not interact in the case of cytochrome c4. PMID- 2990551 TI - Internal protein motions, concentrated glycerol, and hydrogen exchange studied in myoglobin. AB - Experiments were carried out to measure the effect of concentrations of glycerol on H-exchange (HX) rates by using myoglobin as a test protein. Concentrated glycerol has only a small slowing effect on the HX kinetics of freely exposed amides, studied in a small molecule model (acetamide). Larger effects occur in structured proteins. The effect of solvent glycerol on different parts of the HX curve of myoglobin was studied by use of a selective "kinetic labeling" approach. Concentrated glycerol exerts an apparently reverse effect on protein H exchange; the faster exchanging "surface" protons are least affected, while the slower and slower amide NH is further slowed by larger and larger factors. These results seem inconsistent with solvent penetration models which generally visualize slower and slower protons as being placed, and undergoing exchange, farther and farther from the solvent-protein interface. On the other hand, the results are as expected for the local unfolding model for protein H exchange since concentrated glycerol is known to stabilize proteins against unfolding. In the local unfolding model, slower exchanging protons are released by way of higher energy and therefore generally larger, unfolding reactions. Larger unfoldings must be more inhibited by the glycerol effect. PMID- 2990553 TI - Preparation of reconstituted cytochrome oxidase vesicles with defined trans membrane protein orientations employing a cytochrome c affinity column. AB - Reconstituted cytochrome oxidase systems in which the majority of the vesicles contain a single oxidase dimer can be prepared. It is shown that, when these are passed through a cytochrome c affinity column, only those vesicles oriented outwards (such that the active site is available to external cytochrome c) are bound to the support matrix. Protein-free vesicles and vesicles containing an inwardly oriented enzyme are eluted in the void volume. Subsequently, vesicles containing an outwardly oriented enzyme can be eluted from the column at high salt concentrations. This protocol has been used successfully to resolve vesicles of either oxidase orientation when the enzyme is reconstituted with a variety of lipid mixtures. The recovery of oxidase activity from the column ranged between 75 and 94%. PMID- 2990554 TI - The role of subunit III in bovine cytochrome c oxidase. Comparison between native, subunit III-depleted and Paracoccus denitrificans enzymes. AB - In order to obtain information on the role of subunit III in the function and aggregation state of cytochrome c oxidase, the kinetics of ferrocytochrome c oxidation by the bovine cytochrome c oxidase depleted of its subunit III were studied and compared with those of the oxidase isolated from P. denitrificans which contains only two subunits. The aggregation state of both enzymes dispersed in dodecyl maltoside was also compared. The two-subunit oxidase from P. denitrificans gave linear Eadie-Hofstee plots and the enzyme resulted to be monomeric (Mr = 82 000) both, in gel filtration and sucrose gradient centrifugation studies. The bovine heart subunit III depleted enzyme, under conditions when the P. denitrificans cytochrome c oxidase was in the form of monomers, was found to be dimeric by sucrose gradient centrifugation analysis. At lower enzyme concentrations monomers were, however, detected by gel filtration. Depletion of subunit III was accompanied by the loss of small polypeptides (VIa, VIb and VIIa) and of almost all phospholipid (1-2 molecules were left per molecule of enzyme). The electron-transfer activity of the subunit III-depleted enzyme showed a monophasic Eadie-Hofstee plot, which upon addition of phospholipids became non-linear, similar to that of the control bovine cytochrome c oxidase. One of the roles of subunit III may be that of stabilising the dimers of cytochrome c oxidase. Lack of this subunit and loss of phospholipid is accompanied by a change in the kinetics of electron transfer, which might be the consequence of enzyme monomerisation. PMID- 2990555 TI - Electron nuclear double resonance of semiquinones in reaction centers of Rhodopseudomonas sphaeroides. AB - Replacement of Fe2+ by Zn2+ in reaction centers of Rhodopseudomonas sphaeroides enabled us to perform ENDOR (electron nuclear double resonance) experiments on the anion radicals of the primary and secondary ubiquinone acceptors (QA- and QB . Differences between the QA and QB sites, hydrogen bonding to the oxygens, interactions with the protons of the proteins and some symmetry properties of the binding sites were deduced from an analysis of the ENDOR spectra. PMID- 2990556 TI - Effects of diamide and dibucaine on platelet glycoprotein Ib, actin-binding protein and cytoskeleton. AB - During extraction of platelets by 1% Triton X-100, the actin-binding protein (platelet filamin) and a 230 kDa protein are degraded by a calcium-activated thiol protease. Occurrence of degradation products of Mr 190 000 (HF-1) and 90 000 (HF-2) is a sensitive indicator of this proteolysis, and can be used to decide whether reduced amounts of the actin-binding protein in extracts are due to proteolysis or to incorporation in the Triton-insoluble (cytoskeletal) fraction. Diamide, which is a sulfhydryl-oxidizing protein cross-linker, inhibits the calcium-activated protease, polymerizes the actin-binding protein and the 230 kDa protein, increases the incorporation of glycoprotein Ib into the cytoskeletal fraction, and inhibits platelet agglutination induced by bovine von Willebrand factor. Inhibition of platelet agglutination by pretreatment with diamide is partly reversed by dibucaine which activates the calcium-activated protease. These observations are in accordance with a working hypothesis that interactions of glycoprotein Ib with cytoskeleton affect, and possibly regulate, its receptor function in the intact platelet. PMID- 2990557 TI - Ribosome-associated cyclic nucleotide-independent protein kinase of Artemia salina cryptobiotic gastrulae. AB - An extra-ribosomal cAMP-independent protein kinase from cryptobiotic embryos of Artemia salina has been purified to near homogeneity by gel filtration on Bio-Gel A-0.5 m, ion-exchange chromatography on DEAE-cellulose and phosphocellulose P11 and affinity chromatography on casein-Sepharose 4B and ATP-agarose. The enzymatic activity has a broad optimum at pH 7-8. Maximal activity is obtained in the presence of 5-6 mM MgCl2. The activity is inhibited by Mn2+, Ca2+ and K+. The enzyme has an Mr of 127 000, utilizes both ATP and GTP as phosphoryl donors and is completely inhibited by heparin and poly(L-glutamic acid). According to its properties, the enzyme can be classified as a casein kinase type II. Although the enzyme is associated with ribosomes, ribosomal proteins are not among the main substrates. The kinase is able to phosphorylate both the alpha and the beta subunits of initiation factor eIF2 using ATP or GTP as phosphoryl donors. The function of phosphorylation in the initiation of protein synthesis is discussed. PMID- 2990558 TI - Genetic expression and gyrase dependence of methylated and undermethylated DNA in Escherichia coli. AB - The genetic expression and dependence on gyrase of plasmid pBR322 were studied in dam-3, dcm-6, and dam-3 dcm-6 derivatives of a minicell-producing Escherichia coli strain. The results obtained with both methylated and undermethylated plasmid DNA were similar. PMID- 2990559 TI - Binary and ternary complexes of malate dehydrogenase with substrates and substrate analogs. AB - Hydroxypyrenetrisulfonate binds to pig mitochondrial malate dehydrogenase (L malate: NAD+ oxidoreductase, EC 1.1.1.37) in the presence and absence of coenzymes with a stoichiometry of one dye molecule/enzyme subunit. Binding is competitive with substrates and known substrate analogs as well as with squaric acid, a newly detected analog forming a ternary complex with enzyme/NAD+ similar to enzyme/NAD+/sulfite. Displacement of hydroxypyrenetrisulfonate by substrates and analogs was used to determine dissociation constants of binary and ternary complexes. Binary complexes form with dissociation constants of about 10 mM. They may be important for kinetic studies at high substrate concentrations where oxaloacetate inhibition and malate activation have been described. PMID- 2990560 TI - Thermal denaturation of cytochrome c peroxidase: pH dependence. AB - Upon heating cytochrome c peroxidase (ferrocytochrome c: hydrogen-peroxide oxidoreductase, EC 1.11.1.5) at pH 4 and 5, the enzyme precipitates at 41 degrees C and 51 degrees C, respectively. Incubating the enzyme at lower temperatures causes a slow dissociation of the heme from the protein. The heme precipitates, while the apoprotein remains soluble. Between pH 6 and 8, the native enzyme is converted to a low-spin ferric form upon heating. The Soret maximum shifts from 408 to 414 nm. The midpoint of this transition is pH-dependent, with a value of 46 degrees C at pH 6 decreasing to 29 degrees C at pH 8. At high temperatures the 414 nm form is converted to a species which has a 'free heme' spectrum with low absorptivity and Soret maximum at 390 nm. The midpoint temperature of this latter transition is 62 degrees C and 57 degrees C at pH 7 and 8, respectively. PMID- 2990561 TI - The molecular species of phosphatidic acid, diacylglycerol and phosphatidylcholine synthesized from sn-glycerol 3-phosphate in rat lung microsomes. AB - The species pattern of phosphatidic acid, diacylglycerol and phosphatidylcholine synthesized from [14C]glycerol 3-phosphate was measured using a newly developed HPLC technique yielding 13 molecular species. A direct comparison of these species patterns presupposes determination of the lipolytic activity of lung microsomes. The lipolytic activity was quantitatively determined by measuring the changes of the endogenous concentration of diacylglycerol, triacylglycerol and free fatty acids. The species pattern of endogenous diacylglycerol measured in the time-course of lipolysis did not show any changes up to an incubation period of 20 min, suggesting that the lipolytic activity showed only a very low selectivity for individual substrate species. Diisopropylfluorophosphate (5 mumol/mg microsomal protein) strongly decreased the lipolytic activities as well as the microsomal phosphatidate phosphohydrolase activity, as measured by means of exogenous phosphatidic acid, and also the generation of phosphatidic acid from [14C]glycerol 3-phosphate. In lung microsomes, labeled phosphatidic acid and diacylglycerols were synthesized from the endogenous free fatty acids and sn [14C]glycerol 3-phosphate, which had previously been added. By addition of CDPcholine to the prelabeled microsomes the synthesis of phosphatidylcholine was measured. After hydrolysis of phosphatidic acid and phosphatidylcholine with cytoplasmatic phosphatidate phosphohydrolase or phospholipase C, respectively, the de novo synthesized species patterns of these two lipids and of the diacylglycerol were determined. Comparison of the species pattern of de novo synthesized phosphatidic acid with that of diacylglycerol largely showed the same distribution of radioactivity among the individual species, except that the relative proportion of label was higher in the 16:0/16:0 and 16:0/18:0 species of phosphatidic acid and lower in the 16:0/20:4 and 18:0/20:4 species than in the corresponding species of diacylglycerol. The species pattern of de novo synthesized diacylglycerol showed no differences from that of the phosphatidylcholine synthesized from it. From this result we concluded that the cholinephosphotransferase of lung microsomes is nonselective for individual species of the diacylglycerol substrate. The 16:0/18:1 and 16:0/18:2 species of phosphatidic acid, diacylglycerol and phosphatidylcholine showed a higher synthesis rate than their 18:0 counterparts, whereas the 16:0 or 18:0 analogues of species containing 20:4 and 22:6 fatty acids showed nearly the same synthesis rates.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990562 TI - Evidence for the existence of isozymes of choline kinase and their selective induction in 3-methylcholanthrene- or carbon tetrachloride-treated rat liver. AB - New evidence is provided that rat liver choline kinase exists in several distinct forms (choline kinases I, II and III) which differ in isoelectric point, molecular size and antigenicity against anti-rat kidney choline kinase IgG. Remarkable and selective induction of the choline kinase II and choline kinase III forms of choline kinase was caused similarly by the administration of polycyclic aromatic hydrocarbon carcinogen, 3-methylcholanthrene or hepatotoxic carbon tetrachloride (CCl4). The immunochemical approach further indicated that the elevation in the activity of choline kinase in the 3-methylcholanthrene- or CCl4-treated rat liver was not accompanied by a parallel increase in the amount of choline kinase II enzyme protein, compatible with the induction of either a small amount of new enzyme protein(s) with very high specific activity or another enzyme which might catalyze post-translational modification of choline kinase. PMID- 2990563 TI - High-affinity incorporation of 20-carbon polyunsaturated fatty acids by human skin fibroblasts. AB - This study has examined the acyl specificity of incorporation of polyunsaturated fatty acids into cellular glycerolipids of human skin fibroblasts. At low exogenous fatty acid concentrations (0.2-1.2 microM) the extent of incorporation of arachidonate, eicosapentaenoate, 8,11,14-eicosatrienoate and 5,8,11 eicosatrienoate is 60-150% greater than that of oleate or linoleate. As the concentration of exogenous free fatty acid is increased to 25 microM, there is little decrease in the percentage of exogenous oleate incorporated into cellular glycerolipids. Under these conditions, the percentage incorporation of arachidonate and eicosapentaenoate drops 2-3-fold and approaches that of oleate. In contrast, the percentage incorporation of 8,11,14-eicosatrienoate remains high as exogenous fatty acid concentrations are increased. Incorporation of arachidonate, eicosapentaenoate, 8,11,14-eicosatrienoate and 5,8,11 eicosatrienoate is inhibited by addition of any of the other C20 polyunsaturated fatty acids but not by palmitate or oleate. C20 polyunsaturated fatty acids other than the eicosanoid precursors are also not effective inhibitors of arachidonate incorporation. The high affinity incorporation of C20 polyunsaturated fatty acids does not appear to be due to their selective esterification in any one class of cellular phospholipids. These results are compatible with a model of two pathways of fatty acids incorporation into mammalian cells. One pathway utilizes all exogenous long-chain fatty acids and, at least in fibroblasts, is not readily saturable. The second is a high-affinity, low-capacity uptake mechanism specific for arachidonate and other precursors of eicosanoids. The acyl specificity of this latter pathway appears to be similar to that of platelet arachidonyl- CoA synthetase. Results obtained with 8,11,14-eicosatrienoate would indicate, however, that at high concentrations, additional mechanisms influence the acyl specificity of fatty acid incorporation in these cells. PMID- 2990565 TI - Phospholipid oxidation catalyzed by cytochrome c in liposomes. AB - Oxidation of liposome phospholipids has been studied in the presence of cytochrome c. Sonicated vesicles of soya bean or egg-yolk lipids, or purified phospholipid preparations, were treated with oxidized cytochrome c at a 10:4 lipid/protein ratio (w/w). Lipid peroxidation was examined by oxygen polarography, gas-liquid chromatography (GLC) and the thiobarbituric acid test. Oxidized, but not reduced, cytochrome effectively catalyzes lipid oxidation under these conditions. Oxygen consumption and disappearance of unsaturated fatty acids follow closely similar patterns, the O2 consumption rate showing a maximum (1.53 mol O2/min per mol heme) shortly before fatty acid loss reaches its peak. GLC and O2 consumption data suggest that monohydroperoxides are the most abundant oxidized species in the system. The thiobarbituric acid reaction, however, appears only to be of qualitative value in peroxidation studies. In order to test the mechanism through which oxidation occurs in our system, the effect of liposome composition and the presence of antioxidants was tested, both on cytochrome c binding to bilayers and on O2 consumption. Oxidized and reduced cytochrome c bind the lipid bilayers with similar affinity, but only the oxidized form is active in autoxidation. Antioxidants do not modify either cytochrome c binding to sonicated liposomes. Lipid composition does influence considerably cytochrome binding, and O2 consumption is correspondingly altered. Studies with various antioxidants and inhibitors suggest that both free radicals and singlet oxygen may be involved in the process under study. PMID- 2990564 TI - The base-exchange enzyme activities of sarcolemma and sarcoplasmic reticulum from rat heart. AB - The Ca2+ dependent incorporation of [14C]ethanolamine, L-[14C]serine and [14C]choline into phosphatidylethanolamine, phosphatidylserine and phosphatidylcholine, respectively, were investigated in membrane preparations from rat heart. The ethanolamine and serine base-exchange enzyme-catalyzed reactions were associated with the sarcolemma and sarcoplasmic reticulum. There was a 17.2-fold and 6.8-fold enrichment, respectively, of the serine and the ethanolamine base-exchange enzyme activities in the sarcolemma compared to the starting whole homogenate. The sarcoplasmic reticulum was enriched in the ethanolamine and serine base-exchange enzyme activities. The choline base exchange enzyme activity of all membranes fractions was negligible compared to the ethanolamine or serine base-exchange enzyme activities. The apparent Km for the ethanolamine and serine base-exchange enzyme in sarcolemma was 14 microM and 25 microM, respectively. The pH optimum for these base-exchange activities was 7.5-8.0. There was a dependence upon Ca2+ for these reactions with a 1 or 4 mM concentration required for maximal activity. The properties of the sarcoplasmic reticulum base-exchange enzymes were similar to the sarcolemmal base-exchange enzymes. PMID- 2990566 TI - Kinetics of phospholipid transfer between liposomes (neutral or negatively charged) and high-density lipoproteins: a spin-label study of early events. AB - The kinetics of spin-labeled phosphatidylcholine transfer between vesicles and HDL particles exhibited a two-phase process, as seen by ESR spectroscopy. The results were analyzed by considering several possible steps in the overall transfer, whose aspects were also studied: (i) micellar complex formation after HDL apolipoprotein-vesicle mixture, (ii) the rate of PC transfer from the micellar complex to HDL, (iii) the rate of the reverse reaction between overloaded HDL particles and other particles such as HDLs, LDLs, and lipid vesicles. The results agree most convincingly with a mechanism in which the diffusion of phospholipids into the HDL-endogenous lipids is the limiting step, occurring as a two-step process. In addition, we observed a negative charge effect on the lipid transfer rates and yields. PMID- 2990567 TI - The capacity of the sarcoplasmic reticulum for phospholipid synthesis: a developmental study. AB - The activities of three enzymes involved in phospholipid synthesis, sn-glycerol-3 phosphate acyltransferase (EC 2.3.1.15), cholinephosphate cytidylyltransferase (EC 2.7.7.15), and cholinephosphotransferase (EC 2.7.8.2), were assayed in adult skeletal muscle. The acyltransferase and cholinephosphotransferase were concentrated in the sarcoplasmic reticulum, where their specific activities were 80 and 33%, respectively, of the specific activity in liver microsomes. Cytidylyltransferase activity was distributed throughout the cell with most of the activity in the cytosol. Its activity in muscle was only 10% of liver activity. Functional sarcoplasmic reticulum was isolated by density gradient centrifugation after calcium loading in the presence of phosphate. The specific activities of these enzymes wee undiminished in the calcium-loaded fraction, suggesting that these enzymes are intrinsic components of the sarcoplasmic reticulum. In developing muscle (2 and 6 days postnatal) acyltransferase and cholinephosphotransferase activities were also present in a calcium-loaded microsomal subfraction at the same level as in the adult. Cytidylyltransferase activity, on the other hand, was 8-fold higher in developing muscle. In addition, developing muscle had a 3-fold increase in the proportion of cytidylyltransferase associated with the microsomal fraction. These data suggest that sarcoplasmic reticulum has the capacity for phospholipid synthesis in mature and developing muscle, and that the rate of phosphatidylcholine synthesis may be regulated by the levels of cytidylyltransferase and by translocation of this enzyme between the sarcoplasmic reticulum and the cytosol. PMID- 2990568 TI - Estrogen and progestin receptors in mouse vaginal epithelium and fibromuscular wall. AB - Both sodium molybdate and Percoll density gradient stabilize the hormone-binding capacities of the estrogen and progestin receptors and individually increase the recovery of these receptors in prepared cytosols of the separated mouse vaginal epithelium and fibromuscular wall. Their effects are additive. The concentrations of estrogen receptors are similar in the epithelial and fibromuscular compartments, whereas progestin receptor concentrations are higher in the epithelium. PMID- 2990569 TI - The effect of detergents on immunoprecipitability of lysosomal sphingomyelinase. AB - Antibodies raised against the soluble form of acid sphingomyelinase from human urine and placenta are able to precipitate about 70% of the sphingomyelinase activity present in preparations of urinary sphingomyelinase. In contrast, no precipitation of sphingomyelinase activity occurs in detergent-containing preparations from placenta or splenic membranes. The formation of immune complexes between the antibodies and urinary sphingomyelinase is inhibited if detergents are added. With the non-ionic detergent Triton X-100 significant inhibition occurs only above the critical micellar concentration of the detergent. With the anionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1 propane sulphonate (Chaps) substantial inhibition is already observed below the critical micellar concentration of the detergent. PMID- 2990570 TI - Effects of semi-dilute actin solutions on the mobility of fibrin protofibrils during clot formation. AB - Low concentrations of actin filaments (F-actin) inhibit the rate and extent of turbidity developed during polymerization of purified fibrinogen by thrombin. Actin incorporates into the fibrin clot in a concentration-dependent manner that does not reach saturation, indicating nonspecific trapping of actin filaments in the fibrin network. Actin does not retard activation of fibrinogen by thrombin, but rather the alignment of fibrin protofibrils into bundles which constitute the coarse clot. In contrast, equivalent F-actin concentrations have little or no effect on the turbidity of plasma clots. The difference is attributed to the presence of a plasma protein, gelsolin, that severs actin filaments. Purified gelsolin greatly reduces the effect of F-actin on the turbidity of a pure fibrin clot and decreases the fraction of actin incorporated by the clot. A calculation of the extent to which the gelsolin concentrations used in these experiments reduce the fraction of actin filaments which are long enough to impede each other's rotational diffusion indicates that it is the overlapping actin filaments which retard the association of fibrin protofibrils. The findings suggest that one role for the F-actin depolymerizing and particularly actin severing activities in blood is to prevent actin filaments released by tissue injury from interfering with the formation of coarse fibrin clots. PMID- 2990571 TI - Effect of glucose on induction of delta-aminolevulinic acid synthase, ferrochelatase and cytochrome P-450 hemoproteins in isolated rat hepatocytes by phenobarbital and lead. AB - In the present work we have been able to demonstrate that phenobarbital and lead exert an inducing effect on the biosynthesis of delta-aminolevulinic acid synthase, ferrochelatase and cytochrome P-450 hemoproteins in isolated rat hepatocytes of normal adult rats. Dibutyryl cyclic AMP enhances the induction effect produced by phenobarbital in this in vitro system. Glucose inhibits the induction of delta-aminolevulinic acid synthase and ferrochelatase. This repression effect can be reversed with increasing concentrations of dibutyryl cyclic AMP. No glucose effect was observed on the phenobarbital- and lead mediated inductions of cytochrome P-450. he present results add more experimental evidence to support the concept that the last enzyme of the heme pathway is inducible, and as such may have a significant role in regulatory mechanisms of porphyrin and heme biosynthesis. PMID- 2990572 TI - Adenylate kinase is a source of ATP for tumor mitochondrial hexokinase. AB - It has been proposed that hexokinase bound to mitochondria occupies a preferred site to which ATP from oxidative phosphorylation is channeled directly (Bessman, S. (1966) Am. J. Medicine 40, 740-749). We have investigated this problem in isolated Zajdela hepatoma mitochondria. Addition of ADP to well-coupled mitochondria in the presence of an oxidizable substrate initiates the synthesis of glucose 6-phosphate via bound hexokinase. This reaction is only partially inhibited by oligomycin, carboxyatractyloside, carbonyl cyanide m chlorophenylhydrazone (CCCP) or any combination of these, suggesting a source of ATP in addition to oxidative phosPhorylation. This source appears to be adenylate kinase, since Ado2P5, an inhibitor of the enzyme, suppresses hexokinase activity by about 50% when added alone or suppresses activity completely when added together with any of the inhibitors of oxidative phosphorylation. Ado2P5 does not uncouple oxidative phosphorylation nor does it inhibit ADP transport (state 3 respiration) or hexokinase. The relative amount of ATP contributed by adenylate kinase is dependent upon the ADP concentration. At low ADP concentrations, glucose phosphorylation is supported by oxidative phosphorylation, but as the adenine nucleotide translocator becomes saturated the ATP contributed by adenylate kinase increases due to the higher apparent Km of the enzyme. Under conditions of our standard experiment ([ADP] = 0.5 mM), adenylate kinase provides about 50% of the ATP used by hexokinase in well-coupled mitochondria. In spite of this, externally added ATP supported higher initial rates of hexokinase activity than ADP. Our findings demonstrate that oxidative phosphorylation is not a specific or preferential source of ATP for hexokinase bound to hepatoma mitochondria. The apparent lack of a channeling mechanism for ATP to hexokinase in these mitochondria is discussed. PMID- 2990573 TI - Partition of divalent and total manganese in organs and subcellular organelles of MnCl2-treated rats studied by ESR and neutron activation analysis. AB - The possibility that Mn2+ is converted to other valency states in vivo was examined by measuring the ratio of Mn2+, determined by ESR, to total manganese, determined by neutron activation analysis combined with chemical separation, in various organs of control rats and rats treated with MnCl2. In control rats, the total manganese content was high in the thyroid, hypophysis, adrenal, pancreas, liver and kidney, but the Mn2+ contents of these organs were low. In rats treated with Mn2+, the total manganese contents of all organs increased, but the Mn2+ contents still remained low. With regard to subcellular distribution, the total manganese content was high in the nuclear and mitochondrial fractions of the liver and kidney, and in the microsomal and supernatant fractions of the pancreas. The ratio of Mn2+ to total manganese was relatively high in the microsomes of the liver and kidney of control rats, and in the nuclear fraction of the pancreas of Mn2+-treated rats. Thus, the distribution and behavior of manganese in the pancreas were different from those in other organs. Purified liver nuclei and mitochondria were demonstrated to contain manganese, indicating that manganese is tightly bound in each cellular compartment. PMID- 2990574 TI - Effect of physical training on glucose transporters in fat cell fractions. AB - Physical training increases maximally insulin-stimulated glucose assimilation and 3-O-methylglucose transport in epididymal fat cells. In the present report, glucose-inhibitable cytochalasin B binding in subcellular fractions of epididymal adipocytes was measured to assess changes in number of glucose transporters induced by training. Groups of rats trained by swimming were compared to control groups of the same age, matched with respect to body weight by restricted feeding. It was found that in trained rats the number of glucose transporters in the low density microsome fractions from non-insulin-stimulated fat cells was larger than in untrained rats. In both groups of rats, insulin stimulation of adipocytes decreased the number of glucose transporters in low-density microsomes by about 60% and increased the number of glucose transporters in the plasma membrane fractions. The number of glucose transporters in the plasma membrane fractions from maximally insulin-stimulated fat cells was larger in trained rats than in control rats. [U-14C]Glucose incorporation into lipids varied in proportion to plasma membrane cytochalasin B binding per cell under all conditions tested. The results explain the enhancing effect of training on insulin responsiveness transport of hexose in fat cells. PMID- 2990575 TI - Comparative effects of trypsin, collagenase and mechanical harvesting on cell membrane lipids studied in monolayer-cultured endothelial cells and a green monkey kidney cell line. AB - Experiments are presented in which membrane lipids of endothelial cells in monolayer culture were labelled with [14C]linoleic acid. Approx. 90% of the radioactive label were incorporated into phospholipids. A comparison of various harvesting methods showed that during the disruption of the labelled endothelial cell monolayer, 0.25% trypsin and 0.125% trypsin (+0.01% EDTA) released 650 and 470% more radioactivity, respectively, than did 0.01% collagenase (+0.01% EDTA). Parallel studies were performed on a green monkey kidney cell line. In this case, 0.25% trypsin released 520% more radioactivity than did 0.1% collagenase (+0.01% EDTA), although 0.125% trypsin in the presence of EDTA (0.01%) was much less traumatic than trypsin alone, the released radioactivity being of the same order of magnitude as that for collagenase. Morphological studies on endothelial cell cultures failed to reveal any distinctive differences in surface morphology following the various enzyme treatments. The results suggest that collagenase treatment of endothelial cell monolayers is the least traumatic harvesting or subculturing method as far as the integrity of the lipids in the cell membrane is concerned. PMID- 2990576 TI - Endothelial binding of transferrin in fractionated liver cell suspensions. AB - Several studies using crude liver cell suspensions incubated with labeled transferrin have led to a conclusion that hepatocytes have transferrin receptors. When a visual probe, which permits evaluation of transferrin binding to individual cells, was used, the binding was unexpectedly found to be limited to endothelial cells in liver cell suspensions. Neither hepatocytes nor Kupffer cells contained transferrin receptors. In the present study, we fractionated liver cell suspensions using metrizamide gradients and centrifugal elutriation to obtain hepatocytes, Kupffer cell and endothelial cell fractions of high purity. Incubation of these fractions with 125I- or 59Fe-labeled transferrin led to exclusive binding to endothelial cells but not hepatocytes nor Kupffer cells. Kinetic analysis demonstrated Kd of 1.9 X 10(-7) M, Bmax of 3.1 pmol/10(6) cells per min, corresponding to 2.1 X 10(5) molecules/cell per min. At 4 degrees C, the binding reached a steady-state plateau within 5 min. Comparison of our data with those of previous investigators demonstrates a consistency if we consider that crude liver cell suspensions are contaminated with 2-3% endothelial cells. Thus, the previously reported findings may be entirely due to the contamination of crude liver cell suspensions with a small number of endothelial cells. PMID- 2990577 TI - Atriopeptin II elevates cyclic GMP, activates cyclic GMP-dependent protein kinase and causes relaxation in rat thoracic aorta. AB - Synthetic atriopeptin II, an atrial natriuretic factor with potent vasodilatory effects, was studied in isolated strips of rat thoracic aorta to determine its actions on contractility, cyclic nucleotide concentrations and endogenous activity of cyclic nucleotide-dependent protein kinases. Atriopeptin II was found to relax aortic strips precontracted with 0.3 microM norepinephrine whether or not the endothelial layer was present. Relaxation to atriopeptin II was closely correlated in a time- and concentration-dependent manner with increases in cyclic GMP concentrations and activation of cyclic GMP-dependent protein kinase (cyclic GMP-kinase). The threshold concentration for all three effects was 1 nM. Atriopeptin II (10 nM for 10 min) produced an 80% relaxation, an 8-fold increase in cyclic GMP concentrations and a 2-fold increase in cyclic GMP-kinase activity ratios. Atriopeptin II did not significantly alter cyclic AMP concentrations or cyclic AMP-dependent protein kinase activity. These data suggest that cyclic GMP and cyclic GMP-kinase may mediate vascular relaxation to a new class of vasoactive agents, the atrial natriuretic factors. Similar effects have been observed with the nitrovasodilator, sodium nitroprusside, and the endothelium dependent vasodilator, acetylcholine. Therefore, a common biochemical mechanism of action that includes cyclic GMP accumulation and activation of cyclic GMP kinase may be involved in vascular relaxation to nitrovasodilators, endothelium dependent vasodilators and atrial natriuretic factors. PMID- 2990578 TI - Three secretory rates in human hepatoma cells. AB - It is presently unknown what factors regulate the rate of intracellular transport of secretory proteins. The human hepatoma cell line Hep G2 is highly differentiated and secretes many of the proteins characteristic of normal hepatocytes. The secretion kinetics of nine proteins by Hep G2 cells in culture was investigated using pulse-chase techniques and immunoisolation of proteins with monospecific antibodies. Our results show that the export of nine proteins falls into three discrete kinetic classes: (i) a rapidly secreted class with an intracellular retention half-time of 30-40 min (albumin, fibronectin, alpha fetoprotein and alpha 1-proteinase inhibitor), (ii) an intermediate secreted class with a half-time of 75-80 min (ceruloplasmin, alpha 2-macroglobulin and plasminogen), (iii) and a slowly secreted class with an intracellular retention half-time of 110-120 min (fibrinogen and transferrin). Our findings that there are three distinct kinetic classes of secretory proteins strongly suggests that intracellular transport is selective and that proteins of the same secretory class share structural features which influence their rate of export. PMID- 2990579 TI - Partial characterization of the macrophage factor that stimulates fibroblasts to produce collagenase and to degrade collagen. AB - Rabbit bone marrow-derived macrophages in culture produce and release a soluble factor that activates collagenase secretion and collagen degradation by cultured skin fibroblasts from either rabbit, mouse or human origin. The factor is heat labile and is inactivated by phenylglyoxal. After gel filtration, it is recovered in both an apparent high-Mr (67000-76000) and a low-Mr (9000-14000) form. Chromatography on cation exchangers suggests two molecular species with different charge properties. These characteristics are compatible with known properties of rabbit interleukin 1. PMID- 2990580 TI - The developmental regulation of L-ornithine decarboxylase in Dictyostelium discoideum and its induction by cAMP. AB - By the use of an in vivo assay, ornithine decarboxylase (L-ornithine carboxy lyase, EC 4.1.1.17) is shown to be developmentally regulated in Dictyostelium discoideum. High levels of cAMP can induce ornithine decarboxylase activity in preaggregative cells kept in shaking suspension, under similar conditions as where other markers for development can also be induced. This induction by cAMP is solely dependent on the total amount of cAMP to which the cells have been exposed, and not on the manner of cAMP addition. Induction of ornithine decarboxylase activity, when measured in vitro, is caused by both an increase in total enzyme activity and by a proportional increase in activity of the high affinity form for the cofactor pyridoxal phosphate. When measured in vivo, an additional regulatory mechanism seems to be involved. Kinetic studies with the competitive inhibitor putrescine suggest that in cAMP-stimulated cells the low affinity form of the enzyme may also be active in vivo. PMID- 2990581 TI - Activation-associated alterations in neutrophil pyridine nucleotide levels: a potential regulatory role for calcium and calmodulin. AB - The concentration of NADP + NADPH in resting human neutrophils has been measured to be 24.0 +/- 2.7 X 10(-18) mol/cell. Upon activation with opsonized zymosan A, phorbol myristic acetate or N-formylmethionyl-leucylphenylalanine, neutrophil NADP + NADPH pools increase to 80.5, 84.0, and 54.0 X 10(-18) mol/cell, respectively. These increases in pyridine nucleotide concentration are blocked by the addition of the calcium antagonist 8-(N,N-dimethylamino)-octyl-3,4,5 trimethoxybenzoate hydrochloride, while calcium ionophore A23187, in the presence of calcium, will trigger the increase in the absence of other stimuli. Calmodulin antagonists trifluoperazine and N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide also inhibit stimulus-induced increases in the NADP + NADPH pool. These studies are interpreted as suggesting a role for calcium and calmodulin, and possibly protein kinase C in the regulation of pyridine nucleotide concentration in the activated neutrophil. PMID- 2990582 TI - Polyamine uptake by bovine adrenocortical cells. AB - Bovine adrenocortical cells of fasciculo-reticulata origin in primary culture actively accumulate polyamines from the extracellular medium in an energy dependent process. At low extracellular concentration (e.g., 1 microM putrescine), the transport system resulted in a several-hundred-fold concentration of polyamine in the cellular compartment within 1-2 h of incubation. Putrescine uptake appeared to be the sum of a sodium-dependent, saturable process, with an apparent Km of about 10 microM and of a non-saturable, sodium-independent component. By contrast, spermine was taken up by the cells mostly in a sodium-independent manner. Cross-competition experiments suggested that both polyamines were at least partly transported by the same system. Using specific corresponding probes, it was shown that the polyamine uptake was independent of the amino acid transport systems of the A, L and N types known in a number of cell systems. Adrenocortical cell polyamine content is known to be modulated by adrenocorticotropin through induction of ornithine decarboxylase activity. The existence of a specific uptake system in these cells opens the possibility of a more rapid pathway for the regulation of cellular polyamine levels. It remains to be examined whether this polyamine transport system is under hormonal control, and whether this can support the suggestion that polyamines may represent a form of intracellular messengers in the mechanism of hormone action. PMID- 2990583 TI - Autocrine role of estrogens in the augmentation of luteinizing hormone receptor formation in cultured rat granulosa cells. AB - The effects of estrogens on gonadotropin-stimulated luteinizing hormone (LH) receptor formation were examined in primary cultures of rat granulosa cells. Granulosa cells were cultured for 3 days with increasing concentrations of follicle-stimulating hormone (FSH) in the presence or absence of native and synthetic estrogens. Follicle-stimulating hormone stimulated LH receptor formation in a dose-dependent fashion, and estrogens enhanced the FSH-stimulated LH receptor content by decreasing the apparent ED50 of FSH. At 6.25 ng/ml FSH, the enhancement in LH receptor was estrogen dose dependent, with an ED50 value of about 3 X 10(-9) M for 17 beta-estradiol. The increased LH receptor content seen in cells treated with FSH and estrogen was correlated with increased cAMP production by these cells in response to LH stimulation. Time course studies revealed enhancement of FSH-stimulated LH receptor induction at 48 and 72 h of culture. Granulosa cells were also cultured with FSH for 2 days to induce functional LH receptors, then further cultured for 3 days with LH in the presence or absence of estrogens. At 30 ng/ml LH, increasing concentrations of estrogens maintained LH receptor content in a dose-dependent fashion, with their relative estrogenic potencies in keeping with reported binding affinities to estrogen receptors. An autocrine role of estrogens on LH receptor formation was further tested in granulosa cells treated with FSH and an aromatase substrate (androstenedione) to increase estrogen biosynthesis. Cotreatment with semipurified estrogen antibodies partially blocked the FSH stimulation of LH receptors, whereas nonimmune serum was ineffective. Also, inclusion of diethylstilbestrol prevented the inhibitory effect of the estrogen antibodies. Thus, local estrogens in ovarian follicles may play an autocrine role in granulosa cells to enhance LH receptor formation and to increase granulosa cell responsiveness to the LH surge, with subsequent ovulation and adequate corpus luteum formation. PMID- 2990584 TI - Isolation and characterization of Sertoli cell plasma membranes and associated plasminogen activator activity. AB - Plasma membranes were isolated from the cultured Sertoli cells of 20-day-old rat testes by differential centrifugation and sucrose density fractionation. The distribution and purity of subcellular components was determined by marker enzyme analysis of gradient fractions. The plasma membrane fraction showed an enrichment in two plasma membrane marker enzymes, 5'-nucleotidase and ouabain-sensitive Na+/K+-ATPase-specific activities, of 9- and 23-fold, respectively. Forty-two percent and 52% of the total cellular 5'-nucleotidase and ouabain-sensitive Na+/K+-ATPase activities, respectively, were found in the membrane fraction. The protein yield of plasma membrane was approximately 6% of the total cellular protein. Two-dimensional polyacrylamide gel electrophoresis was used to compare [35S] methionine- and [3H] glucosamine-labeled membrane proteins. The incorporation of [35S] methionine and [3H] glucosamine was increased in several proteins when the cultured Sertoli cells were treated with follicle-stimulating hormone, insulin, retinol, and testosterone. Isolated Sertoli cell membranes contained a membrane-associated form of plasminogen activator. Analysis of this plasminogen activator demonstrated that the membrane-associated enzyme existed primarily as a single 38,000-40,000-Mr form. PMID- 2990585 TI - Lipid phase of transverse tubule membranes from skeletal muscle. An electron paramagnetic resonance study. AB - The lipid phase of transverse tubule membrane was probed with a variety of fatty acid spin labels. The motion of the probe increased as the distance between the spin label and polar head group increased, in agreement with results reported in other membranes. The value of the order parameter at 37 degrees C for a fatty acid spin label containing the label attached to its fifth carbon atom was closer to values reported for bacterial membranes than to the lower values reported for other mammalian membranes. Order parameters for spin labels containing the label nearer to the center of the bilayer were closer to the values reported in other mammalian membranes than to values reported for bacterial membranes. These results indicate that the lipid segments in the vicinity of the polar head group, and less so those near the center of the bilayer, are motionally more restricted in transverse tubules than in other mammalian membranes. In particular, the lipid phase of the transverse tubule membrane is less fluid than that of the sarcoplasmic reticulum membrane. A possible role of the high cholesterol content of transverse tubules in generating the lower fluidity of its lipid phase is discussed. PMID- 2990587 TI - Thermotropic behavior of dimyristoylphosphatidylcholine in the presence of cytochrome c oxidase. PMID- 2990586 TI - The inhibition of muscle contraction by adenosine 5' (beta, gamma-imido) triphosphate and by pyrophosphate. AB - We have studied the inhibition of the contraction of glycerinated rabbit psoas muscle caused by ligands that bind to the ATPase site of myosin. Two ligands, adenosine 5' (beta, gamma-imido) triphosphate (AMPPNP) and pyrophosphate (PPi), decreased the force and stiffness developed in isometric contractions and the velocity of shortening of isotonic contractions. The force exerted by isometric fibers was measured as a function of MgATP in the presence and absence of a constant concentration of the ligands. As the MgATP concentration decreased, the inhibition of tension caused by the ligand increased, reaching approximately 50% at 25 microM MgATP and either 2 mM MgPPi or 2 mM MgAMPPNP. The maximum velocity of shortening was also measured as a function of MgATP concentration in the presence of 1 and 2 mM MgPPi and 2.5 and 5 mM MgAMPPNP. Both ligands acted as pure competitive inhibitors with Ki = 3.0 mM for PPi and 5.1 mM for MgAMPPNP. These data show that both ligands are weak inhibitors of the contraction of fibers. The results provided information on the energetics of actin-myosin-ligand states that occur in the portion of the cross-bridge cycle where MgATP binds to myosin. A simple analysis of the inhibition of velocity suggests that MgAMPPNP binds to the actomyosin complex at this step of the cycle with an effective affinity constant of approximately 2 X 10(2) M-1. PMID- 2990588 TI - Polymerized human serum albumin binding activity of sera from patients with various liver disorders (studied by a passive hemagglutination test). PMID- 2990589 TI - The Epstein-Barr virus (EBV): a Rosetta Stone for understanding the role of viruses in immunopathological disorders and in human carcinogenesis. AB - The Epstein-Barr virus (EBV), a B lymphotropic virus, is involved in a growing number of immunopathological disorders benign or malignant. The X-linked lymphoproliferative syndrome and its multifaceted clinical expression in a unique situation described in this issue by Purtilo. Among recent findings, the association between EBV and idiopathic interstitial pneumopathy (also named cryptogenic fibrosing alveolitis), is to be noted (6). From a molecular biology view-point, in vitro immortalization of B lymphocytes by EBV is under a pluri genic (EBNA 2, EBNA 1, LYDMA) control. The role of EBV in oncogenesis appears different in Burkitt's Lymphoma (BL) and in nasopharyngeal carcinoma (NPC). In development of African BL, EBV appears to initiate the multistage carcinogenic event, through an early and massive infection. Other events include specific depression of T-cell immunity by hyperendemic malaria and c-myc onc-gene activation through chromosome translocation. In the genesis of NPC, the role of EBV still remain to be clarified although the strong and consistent association between EBV and the undifferentiated carcinoma of the nasopharyngeal (NPC) around the world favours an etiological relationship. The simple detection of IgA antibodies to VCA and EA allows early detection of the NPC, thus permitting a 95% cure rate at 5 years post-radiotherapy. Such an early diagnostic is of paramount public health importance. Furthermore, IgA/VCA and IgA/EA antibodies characterize precancerous conditions, giving the theoretical possibility of preventive interventions. PMID- 2990590 TI - Epstein-Barr virus-induced diseases in the X-linked lymphoproliferative syndrome and related disorders. AB - Information regarding the development of diverse diseases associated with EBV virus in immune deficient patients has been gained by studying males with XLP, and their families. Multiple immune defenses normally protect against the ubiquitous EBV. Depending on the type and degree of inherited or acquired immunodeficiency, EBV may more or less be capable of inducing a variety of diseases. Multiple methods may be needed to document EBV in the immune deficient individual. Rational approaches to prevention and intervention in EBV-induced diseases in immune compromised individuals are being developed. PMID- 2990591 TI - The molecular biology of Epstein-Barr virus. AB - Epstein-Barr virus (EBV) was discovered in continuously growing tumor cells derived from African patients with Burkitt's lymphoma. In the intervening twenty years, much biological and biochemical information has been accumulated. The virus infects B lymphocytes and occupies a unique position among human herpesviruses in that it is the only one which is capable of forming a latent infection whereby complete copies of the virus genome persist in growth transformed cells. Since there are no fully permissive cell systems of virus replications, only the established B cell lines are available for study of the molecular events of EB virus in infected cells. A viral cycle consists of four stages, latent, early replicative, middle replicative and late replicative stages. In the latent state, only small parts of the viral genome are transcribed and express transformation proteins: nuclear antigens (EBNAs) and lymphocyte determined membrane antigen (LYDMA). After reactivation of viral genome and during a productive cycle, more than 50 RNAs are expressed and over 30 viral specified polypeptides are detectable by immunoprecipitation with a high titer human anti-EBV serum. During the early replicative stage, early antigens (EA) and DNA enzymes, both necessary for DNA synthesis, are synthesized. In the late replicative stage, about 30-40 mRNA are transcribed and two major late antigen complexes, viral capsid antigens (VCA) and membrane antigens (MA), are identified. These antigens are indispensable for the formation of virions. PMID- 2990592 TI - Water exchange through erythrocyte membranes: p-chloromercuribenzene sulfonate inhibition of water diffusion in ghosts studied by a nuclear magnetic resonance technique. AB - A comparison of water diffusion in human erythrocytes and ghosts revealed a longer relaxation time in ghosts, corresponding to a decreased exchange rate. However, the diffusional permeability of ghosts was not significantly different from that of erythrocytes. The changes in water diffusion following exposure to p chloromercuribenzene sulfonate (PCMBS) have been studied on ghosts suspended in isotonic solutions. It was found that a significant inhibitory effect of PCMBS on water diffusion occurred only after several minutes of incubation at 37 degrees C. No inhibition was noticed after short incubation at 0 degree C, as previously used in some labelling experiments. This indicates the location in the membrane interior of the SH groups involved in water diffusion across human erythrocyte membranes. The nuclear magnetic resonance (n.m.r.) method appears as a useful tool for studying changes in water diffusion in erythrocyte ghosts with the aim of locating the water channel. PMID- 2990594 TI - In vitro establishment of human fibroblasts of lysosomal diseases, GM1 gangliosidosis and Sandhoff disease, by transformation with origin-minus SV40 DNA. AB - The permanent human cell lines preserving defects of lysosomal enzymes, GM1-1019 SV and SA-1077-SV, were established from the respective fibroblasts from patients with GM1-gangliosidosis and Sandhoff disease by transfection with replication origin-minus simian virus 40 DNA. These cells grow rapidly without entering senescence during more than 120 population doublings. The activity of beta galactosidase in GM1-1019-SV and of beta-N-acetylhexosaminidase in SA-1077-SV was respectively 40- and 180-fold lower than that of normal fibroblasts. PMID- 2990593 TI - Short term regulation of acyl CoA: cholesterol acyl transferase (ACAT) activity in the regenerating and perinatal liver. AB - Acyl coenzyme A : cholesterol acyltransferase (ACAT), the enzyme catalyzing the hepatic cholesterol esterification, could be involved in the modified availability of cholesterol detectable in proliferating systems. While no significant variations are detectable in the regenerating liver, the modified ACAT activity during liver development and its differential sensitivity to the in vitro stimulation of modulatory systems suggest an involvement of the enzyme in this proliferating process. PMID- 2990595 TI - [Specific binding of 3H-naloxone with isolated rat enterocytes]. AB - Specific 3H-naloxone binding with isolated rat enterocytes has been demonstrated. This binding was selectively inhibited by different opiates and proved to be irreversible, temperature-dependent and sodium azide-sensitive. The presence of naloxone binding sites on enterocytes indicates a possible involvement of opiates in the regulation of enterocyte functions. PMID- 2990596 TI - [Restriction map of the genetic transfer factor pAP42]. AB - Based on the calculated molecular weights of EcoR1, HindIII, and SalI fragments of the genetic transfer factor pAP42 the restriction map of this plasmid was designed. Sites recognizing restrictases are mostly located in the plasmid fragment with a molecular weight of 5.7 MD. PMID- 2990597 TI - [Neurospecific proteins in human malignant brain tumors]. AB - The content of neurospecific proteins S-100, GFA and D2 was measured in malignant cerebral tumors by electrophoresis with the use of monospecific antisera. Concomitant measurement of proteins S-100 and GFA is a more reliable diagnostic criterion as to the tumor histogenesis than study of each protein alone. D2 protein appeared to be the most stable specific marker. PMID- 2990598 TI - [Immunomorphological research on the changes in the focal contacts of cultured cells transformed by Kirsten sarcoma virus]. AB - Alterations of focal contacts (FC) of NRK cultured cells transformed by Kirsten's sarcoma virus and by ts-mutant of this virus were examined immunomorphologically using a new antibody-exclusion method. In nontransformed NRK cells the FC are arranged in the form of a network near the cell edges; numerous single contacts are seen in the center. The FC are always present in both KNRK and tsKNRK cells at permissive temperature. However, their quantity is drastically reduced, especially in the center of the cells, as is often their size. It is suggested that expression of the product of viral oncogene interferes with maturation of the FC continuously formed at the cell edge. PMID- 2990599 TI - [Effect of thymalin on the function of the blood kallikrein-kinin system in thymectomized rats]. AB - The effects of thymus ablation and injection of thymalin on the blood plasma kallikrein-kinin system were studied. Thymus ablation was followed by activation of kinin formation, evidenced by an elevation of the total kallikrein activity and drop of the kininogen level. Injection of thymalin into thymectomized animals makes the characteristics under study return to normal. PMID- 2990600 TI - [Effect of alpha-tocopherol on the adrenal reaction to cold stress]. AB - Adrenocortical function and the intensity of lipid peroxidation in the liver of animals fed the diet containing alpha-tocopherol (4 mg/day/rat for 7 days) and exposed to cold stress for 2, 5 and 20 hours were studied in vitro. It was established that alpha-tocopherol reduces the duration of the corticosterone secretion augmentation under cold stress but raises the sensitivity of the adrenals to ACTH. The action of the antioxidant correlated with its content in the tissues and inhibition of the cold-induced activation of lipid peroxidation. PMID- 2990602 TI - [Cytoplasmic cAMP receptors in the kidneys of rats of different ages studied by gel filtration]. AB - Cytosol reception of cAMP was explored in the kidneys of Wistar rats aged 15 and 60 days. It was shown by gel filtration on Sephadex G-100 that elevation of the cAMP concentration to 10(-9) and 10(-8) brought about a decrease in the molecular weight of receptor complexes, with this effect being more potent in adult animals. Scatchard's analysis of receptor affinity to cAMP demonstrated that in adult animals, the given parameter was considerably less than in young ones. It is assumed that the age-associated changes in cAMP reception are linked with the development of renal sensitivity to the action of hormones. PMID- 2990601 TI - [Mechanisms of the disorder of the hormonal regulation of plasma membrane adenylate cyclase in the rat liver during early ontogeny under in vitro irradiation]. AB - Possible reasons for the previously detected inhibitory effect of gamma-radiation (100-1000 Gy in vitro) on isoproterenol-stimulated activity of adenylate cyclase (AC) of the liver plasma membranes of a 20-day-old rat fetus were investigated. It was discovered that the number of beta-adrenergic receptors was reduced. Analysis of basal and fluorine-stimulated activities of AC attests to the radiation-induced damage to catalytic and N-regulatory proteins. The radiation induced derangement of membrane lipids, where AC functions, is shown. PMID- 2990603 TI - [Changes in purine metabolism in the macrophages of mice exposed to a new synthetic analog of muramyl dipeptide]. AB - It has been established that N-acetylglucosaminyl-N-acetylmuramyl-L-alanyl-D isoglutamine (GMDP), a new synthetic analog of muramyl dipeptide, while incubated in vitro with macrophages essentially inhibits 5'-nucleotidase (5-N) activity without any influence on the activity of adenosine deaminase in these cells. The maximal effect was recorded 24 h after co-incubation. As 0.01 = 1 microgram/ml concentration of GMDP was added, the enzyme activity gradually decreased to minimum. L-D-isomer of GMDP was shown to affect 5-N activity whereas the effect of its analog with a double peptide chain GM (DP)2 was found to be less. Inhibition of 5-N activity may be one of the mechanisms by which macrophages are activated under the influence of GMDP. PMID- 2990604 TI - [Effect of the antioxidant dibunol on the electron paramagnetic resonance signals in the tissues of rats of different ages]. AB - Changes in the intensity of EPR signals from the skeletal and heart muscles, liver, kidneys, thyroid, adrenal cortex, and blood were studied in relation to the animals age and the effect of the antioxidant dibunol. Essential changes were revealed in metabolic activity of the majority of the tissues under study as early as within the first 6 hours after antioxidant administration. PMID- 2990605 TI - [Effect of lithium chloride on thyroid structural elements in the rat and on the balance of calcitropic hormones]. AB - Chronic experiments were performed to study the effect of lithium chloride on morphological and functional characteristics of rat thyroid and on the endocrine regulation of calcium metabolism. As a result of a prolonged exposure to low doses of lithium the thyroid manifests the signs of activation of tissue structures. As the lithium dose is raised, the thyrostatic effect of lithium occurs according to the mechanism similar to the Wolff-Chaikoff effect (colloid accumulation by follicles and suppression of hormones secretion into blood). Lithium chloride stimulates calcitonin production by C cells of the thyroid. However, the increase of the basal level of calcitonin has no substantial effect on calcium metabolism in the animals, since such an increase is accompanied by increment of the production of the physiological antagonist parathyroid hormone by the secretory cells of the parathyroid glands. PMID- 2990606 TI - [Expression of antigens immunologically related to the antigens of the mouse mammary tumor virus in the tissue of human breast tumors]. AB - Indirect radioimmunoblotting on nitrocellulose filters and ELISA on polystyrene plates were employed to test the specimens of the normal and tumor tissue of the mammary gland for antigens which cross react with antigens of structural proteins of mouse mammary gland carcinoma virus. The antigens were detected in 9 tumor specimens out of 25 under testing and in 1 out of 15 specimens of the normal tissue. Such antigens failed to be detected in fibroadenoma or tumors of other sites. PMID- 2990607 TI - [Autoradiographic study of the ontogeny of the imipramine receptors in the cerebral cortex of rats]. AB - Localization of 3H-imipramine binding sites in the cerebral cortex of rats was studied on the 19th day of prenatal development and on the 3d and 14th days of postnatal ontogenesis. Imipramine was injected into rats on days 17, 18 and 19 of gestation. The maximal imipramine receptor density was found in cortex layers IV V. The therapeutic doses of imipramine provoked marked stimulation of the formation of imipramine receptors on the 3d day of postnatal development. However, on the 14th day the 3H-imipramine binding level was stabilized. PMID- 2990608 TI - Associated chronic lymphocytic leukemia and multiple myeloma: origin from a single clone. AB - We investigated the clonal relationship of malignant cells in a patient affected with both chronic lymphocytic leukemia (CLL) and multiple myeloma (MM). CLL cells and malignant plasma cells synthesized IgG1 kappa and IgA kappa molecules, respectively; these monoclonal Ig shared idiotypic determinants, providing evidence that a single clonal disease occurred in this patient. Furthermore, when leukemic CLL cells were driven to differentiate in vitro to immunoblasts and plasma cells, a switch from IgG to IgA occurred in a significant percentage of cells that were double producers. These data suggest that, in some circumstances, CLL leukemic B cells may reach a more mature state, leading to the occurrence of clinical MM. PMID- 2990609 TI - Prolymphocytic leukemia of B cell type: rearranged immunoglobulin (Ig) genes with defective Ig production. AB - An unusual case of prolymphocytic leukemia of the B cell type (B-PLL) in a 79 year-old patient is reported. The clinical and cytomorphological features of the disease were typical of B-PLL, but membrane and cytoplasmic immunoglobulins (Ig) could not be demonstrated by immunofluorescence techniques; 3% to 4% of the cells were shown to have IgG kappa in the cytoplasm by a more sensitive immunoperoxidase method. The cells were unreactive with a panel of monoclonal antibodies against T cell antigens but they were positive with B cell lineage reagents: FMC4, anti-HLA-Dr determinants; FMC7, which reacts with most B-PLL; anti-B1 and anti-B4, which react with most B cell leukemias. Analysis of Ig genes at the DNA level demonstrated that both heavy-chain alleles and one kappa chain allele were rearranged, confirming that the patient's cells were of B lineage. Chromosome analysis revealed a consistent abnormality, t(17;21)(p11;p11), in all cells and, in addition, a 14q+ marker in 10% of the cells. This study highlights the value of DNA analysis techniques for the characterization of neoplastic B cells. The low rate of expression of Ig genes, despite their rearrangement, suggests that a specific transcriptional or posttranscriptional defect must exist in these cells. PMID- 2990611 TI - Recovery of T cell subsets after autologous bone marrow transplantation is mainly due to proliferation of mature T cells in the graft. AB - In 22 patients with malignancies, treated with high-dose chemoradiotherapy and autologous bone marrow transplantation (BMT), peripheral blood T cell subsets and functions were studied. In ten cytomegalovirus (CMV)-negative patients, CD4+ and CD8+ T cells (representing T cells of the helper/inducer phenotype and T cells of the suppressor/cytotoxic phenotype, respectively), recovered slowly and simultaneously. In 12 CMV-positive patients, however, CD8+ T cells recovered more rapidly than CD4+ T cells and rose to increased counts. No T cells with an immature phenotype (CD1+, OKT6+) were observed. Lymphocyte stimulation by herpes simplex virus infected fibroblasts (and by CMV-infected fibroblasts in CMV positive patients) in contrast remained high and even increased after BMT in both groups. These data indicate that T cell recovery after autologous BMT is mainly due to proliferation of mature T cells present in the BM graft and not to generation of new T cells from T cell precursors. PMID- 2990612 TI - A fruit fly bioassay with phosphotriesterase for detection of certain organophosphorus insecticide residues. PMID- 2990610 TI - Constitutive production of leukemia differentiation, colony-stimulating, erythroid burst-promoting, and pluripoietic factors by a human hepatoma cell line: characterization of the leukemia differentiation factor. AB - Conditioned medium (CM) obtained from a human hepatoma cell line, SK-HEP-1, contains colony-stimulating factors (CSFs) active on murine and human bone marrow derived granulocyte and macrophage colony-forming units (CFU-GM) and a factor capable of inducing granulocyte-macrophage differentiation (GM-DF) of murine myelomonocytic leukemic cells WEHI-3B(D+) and human promyelocytic leukemic cells HL-60 when assayed in semisolid agar cultures. The human active granulocyte macrophage colony-stimulating factor (GM-CSF) for day 7 CFU-GM and the GM-DF for WEHI-3B(D+) and for HL-60 are not separable by acrylamide agarose column chromatography, eluting at an apparent molecular weight between 20,000 and 35,000 daltons, or by isoelectric focusing (isoelectric point, pH 5.4). In addition, SK HEP-1 CM contains erythroid burst-promoting activity (BPA) and a factor that promotes the growth of human mixed colonies. SK-HEP-1 cells, which grow as an adherent monolayer, appear not to be endothelial or monocytic in origin since by immunofluorescent staining they are negative for Ia (HLA-DR), monocyte antigen 1 and 2, lysozyme, and factor VIII-related antigen. Positive immunofluorescent staining for keratin and fibronectin suggests the possibility that SK-HEP-1 is an epithelial cell line. Constitutive production of GM-DF as well as other hematopoietic activities including GM-CSF, erythroid BPA, and an activity that promotes the growth of human mixed colony progenitors by a human epithelial tumor cell line, SK-HEP-1, suggests that this cell line is a valuable resource for both large-scale production of these factors and the cloning of the gene(s) that code for these regulators. PMID- 2990613 TI - Reflections on complex problems of the hip in the adolescent. PMID- 2990614 TI - Observations on the use of computerized axial tomography in the management of congenital dislocation of the hip. AB - CAT scans used postreduction to evaluate possible complications or problems in children treated for congenitally dislocated hips can provide special data regarding the adequacy of reduction. PMID- 2990615 TI - Current concept: the expandable prosthesis. PMID- 2990616 TI - Osteosarcoma: a report on past and current treatment methods. PMID- 2990617 TI - Diagnostic and operative arthroscopy of the ankle technique and indications. AB - The technique of ankle arthroscopy is outlined and cases are presented illustrating some of its current applications. The author believes that ankle arthroscopy can play a valuable role in both the evaluation and treatment of traumatic, degenerative, and inflammatory disorders of the synovial and articular linings of the joint. PMID- 2990618 TI - A 30-year follow-up and failure analysis of a Fixel acrylic femoral prosthesis. AB - An acrylic-headed prosthesis was removed after 30 years and analyzed for material deterioration. Although the acrylic head showed some wear and minor corrosion of the cobalt-chromium stem was found, tissue responses were not significant, indicating that use of these materials is consistent with the long-time survival of a prosthesis. PMID- 2990619 TI - The troublesome carpal bones. PMID- 2990620 TI - Fractures of the radial head in adults with the complication "cubitus valgus". AB - Cubitus valgus is a well-known complication of fracture of the bones of the elbow in childhood, but it is an uncommon deformity in adults. Three adult patients are presented, in each of whom cubitus valgus developed as a complication of a fracture and resection of the radial head. The importance of reestablishing the integrity of the ligaments of the elbow following severe injuries to the elbow is stressed. PMID- 2990621 TI - The value of carpal tunnel trispiral tomography in the diagnosis of fracture of the hook of the hamate. AB - Fractures occurring at the base of the hook of the hamate are difficult to detect. Failure to visualize the entire hamulus on routine radiographs and conventional carpal tunnel view may contribute to a delayed diagnosis. Carpal tunnel trispiral thin-section tomography is a useful aid in the investigation of a suspected fracture of the hook of the hamate. PMID- 2990622 TI - Aneurysmal bone cyst of the fibula: en bloc resection with allograft reconstruction. AB - Two cases are presented in which allograft reconstruction followed wide resection of recurrent, aggressive aneurysmal bone cysts of the distal fibula. The authors find that allograft implantation provides a safe and effective method of reconstruction. PMID- 2990623 TI - Giant cell tumor of the tendon sheath with a pathologic phalangeal fracture. A rare association. AB - Giant cell tumor of the tendon sheath is a soft tissue tumor which sometimes affects the adjacent bone structure, producing erosions. In certain rare instances it may manifest as a pathologic fracture. PMID- 2990624 TI - Lack of knowledge of the lymphatic system. PMID- 2990625 TI - Epstein-Barr virus transformed lymphoid cell lines as a new model system in culture for the study of GM2-gangliosidoses: Tay-Sachs and Sandhoff diseases. AB - Epstein-Barr Virus transformed cell lines (LCL) were established from blood B lymphocytes of patients affected with GM2-gangliosidoses variant O (Sandhoff disease, SD) and variant B (Tay-Sachs disease, TSD). LCL from SD showed a severe deficiency of activity of the major lysosomal beta-N-acetylhexosaminidase isoenzymes, Hex A and B; the residual activity was due to Hex S and Hex C. In LCL from TSD, the whole Hex activity was not deficient but isoenzyme composition was completely abnormal. Ultrastructural investigations showed the presence of pleiomorphic enlarged lysosomes appearing as clear vacuoles containing a finely fibrillo-granular material characteristic of the visceral lysosomal storage of gangliosidoses. PMID- 2990626 TI - Cytomegalovirus and schizophrenia. A test of a viral hypothesis. AB - Cerebrospinal fluid (CSF) from 20 chronically hospitalised male schizophrenics and from 12 patients admitted with acute schizophrenia were examined for antibodies against cytomegalovirus. A sensitive and specific enzyme-immunoassay was used to detect IgG or IgM classes of antibodies in the CSF of the schizophrenic patients and of ten orthopaedic patients, who served as controls. No significant amounts of IgM antibody were found in the CSF of either group. A significant titre of IgG was found in only one of the 32 schizophrenics, an acute patient, but in four of the orthopaedic patients. The results do not support an association of cytomegalovirus infection with schizophrenia; if such an association occurs, it must be unusual. PMID- 2990627 TI - Tritiated etorphine and naloxone binding to opioid receptors in caudate nucleus in schizophrenia. AB - Opioid receptor binding sites were assessed in membrane preparations of caudate nucleus from post-mortem brains of controls and of patients with schizophrenia. There was no difference between the two groups in the total specific binding of 3H-etorphine or in its 'mu' and ('delta + kappa') components. Similarly, the binding of 3H-naloxone did not differ between patients and controls. It is concluded that a previous report of reduced opioid receptors in caudate of schizophrenics is unlikely to prove a consistent finding and that the results of the present study offer no support to the claim that there is a general disturbance in opiate mechanisms in schizophrenia. PMID- 2990628 TI - Radio-isotopic joint scans in haemophilic arthritis. AB - The majority of severe haemophiliacs will develop a crippling arthritis consequent upon recurrent haemarthroses although the pathogenic mechanism remains unclear. We have carried out technetium-99 pertechnetate joint scans in the elbows, knees and ankles of 23 haemophilic patients and compared the isotope uptake of the 15 patients with clinical and radiological evidence of arthritis to that of eight patients without arthritis, to 13 age-matched healthy male controls, and to 10 age-matched males with active rheumatoid arthritis. Isotope uptake into the knee joints was significantly higher in haemophilic arthritis than in controls, haemophiliacs without arthritis, and patients with rheumatoid arthritis [median percentage uptake of administered technetium (Tc) dose X 10(3) in right knee = 9.4, 6.2, 6.4 and 6.6, respectively]: and the differences from haemophilic arthritis were all significant (p less than 0.01, Mann-Whitney U test). Similar differences were seen in the elbows and ankles. Increased Tc uptake correlated strongly with frequency of haemarthrosis, pain, synovitis, range of movement and radiological changes in knees and elbows, but poorly with the lesser changes seen in the ankles. These results would support the theory that haemophilic arthritis amongst the inflammatory arthropathies and that scanning is an appropriate technique for following progression of joint disease. PMID- 2990629 TI - Segmental and subsegmental resections of the cirrhotic liver under hepatic inflow and outflow occlusion. AB - Segmental and subsegmental resections of the cirrhotic liver were carried out on 44 patients with hepatocellular carcinoma (HCC) and three cases with regenerating liver nodules. Seventeen patients underwent hepatectomies without hepatic vascular arrest (Group 1). In Group 2 (19 occasions), the Pringle manoeuvre was used for 8-46 min during parenchymal dissection. Hepatic resections were performed during simultaneous occlusion of hepatic inflow and outflow for 20-47 min in 11 patients (Group 3). The estimated blood loss during surgery was 2924 ml in Group 1, 1239 in Group 2, and 765 in Group 3 (Group 1 versus Group 2; P less than 0.02, Group 1 versus Group 3; P less than 0.02). Haemorrhagic shock occurred in three patients of Group 1, one of Group 2, and none of Group 3. Life threatening postoperative complications occurred in four patients in Group 1, two in Group 2, and none in Group 3. Hospital death rate was 23.5 per cent in Group 1, 10.5 per cent in Group 2, and nil in Group 3. The present results may indicate that in performing nonanatomical resection of cirrhotic livers temporary occlusion of both hepatic inflow and outflow can be achieved for at least 30 min and is useful in reducing postoperative morbidity and mortality. PMID- 2990630 TI - Rh(D) haemolytic disease of the newborn: the changing scene. PMID- 2990631 TI - Aminoglutethimide (with hydrocortisone) induced agranulocytosis in primary breast cancer. PMID- 2990632 TI - Muscle contractures associated with glucocorticoid deficiency. PMID- 2990633 TI - "Missed pill" conception: fact or fiction? PMID- 2990635 TI - HTLV-III, haemophilia and blood transfusion. PMID- 2990636 TI - [Effects of lead poisoning on properties of brain mitochondria in young rats]. AB - When the litters were 14 days old, wistar rat pups received lead ions. The suckling mother was fed by a powdered laboratory chow containing lead carbonate (1%, 2%, 4%). The intoxication was going on for 3 days to 9 weeks. While lead-fed pups showed a decrease in body-weight gain and in neurologic development, lead ions were without effects on oxygen consumption, ADP-phosphorylation and activity of several enzymes of cerebral mitochondria. On the other hand, the important lead accumulation into cerebral mitochondria was in a straight ratio with lead doses in the feedings. To explain the in vivo data, the effects of lead acetate were investigated in vitro on brain mitochondria of young male wistar rats. Our findings indicate that lead ions alter mitochondrial respiration and ADP phosphorylation and that inorganic phosphate has a protective effect. PMID- 2990634 TI - IgM and IgG antibodies to human T cell lymphotropic retrovirus (HTLV-III) in lymphadenopathy syndrome and subjects at risk for AIDS in Italy. AB - A study was performed to assess the prevalence of specific antibodies to human T cell lymphotropic retrovirus (HTLV-III) in patients with lymphadenopathy syndrome, patients with the acquired immune deficiency syndrome (AIDS), and those at risk of AIDS. Serum samples were obtained from these groups and from healthy controls in selected cities in Italy, and antibodies to HTLV-III were measured by immunofluorescence assay and, in a few patients, by Western blotting. In addition, IgM antibody values were measured in 82 of those positive for HTLV-III. Altogether, 235 out of 320 patients with lymphadenopathy syndrome had antibodies to HTLV-III, the proportions being highest in haemophiliacs, homosexuals, and drug addicts from Rome; 11 out of 12 patients with AIDS had antibodies; 78 out of 439 subjects at risk for AIDS had antibodies; and six out of 30 patients with lymphadenopathy syndrome and positive for HTLV-III antibodies and nine of 52 patients at risk of AIDS had a detectable titre of IgM. HTLV-III is widespread in groups at risk of AIDS in Italy, and antibodies to HTLV-III are highly prevalent in patients with lymphadenopathy syndrome. A higher proportion of drug abusers were positive for antibodies than in previous studies. HTLV-III "infection" would appear to be spread mainly in compromised hosts, as none of the controls were positive for antibodies. PMID- 2990637 TI - [Study of stereoisomers of N6-phenylisopropyladenosine on the secretion of pancreatic glucagon]. AB - This work was designed to characterize the adenosine receptor (A1 or A2) involved in glucagon secretion. The most potent adenosine analogues on A1 receptors are the N6 substituted compounds, among them N6-phenylisopropyladenosine (PIA); furthermore L-PIA is 50 to 100 times more potent than D-PIA on the A1 receptor, whereas it is 3 to 5 times more potent on the A2 receptor; thus the A1 receptor shows a much higher stereoselectivity. The effects of L-PIA and D-PIA were studied on glucagon secretion from the isolated perfused rat pancreas. 1) L-PIA at 1.65 microM induced a transient glucagon secretion which was not greater than that induced by the same concentration of adenosine. 2) D-PIA at a 3 fold higher concentration (4.95 microM) elicited a secretion of glucagon comparable to that induced by L-PIA 1.65 microM; thus the involved receptor does not present a high stereoselectivity for L-PIA. These results support the fact that the receptor involved in glucagon secretion is not of the A1 type. PMID- 2990638 TI - [Transformation by SV 40 virus sensitizes fibroblasts of human skin to the lytic action of H-1 parvovirus]. AB - Human skin fibroblasts which are naturally resistant to Parvovirus H-1 can be lysed by this virus after SV40 transformation. This observation raises the possibility that oncosuppression by Parvovirus involves a direct oncolytic effect. PMID- 2990639 TI - [Reactivation of gamma-ray irradiated parvovirus H-1 in cells of patients with ataxia telangiectasia and Huntington's chorea]. AB - Parvovirus H-1 was used to probe the cellular radiosensitivity of two human degeneration syndromes AT and HC. No difference in the survival of gamma irradiated H-1 was detected between skin fibroblasts from such patients and from a normal individual. However, AT and normal cells were distinguished by the fact that the reactivation of irradiated H-1 could be increased by UV or X-irradiation of the latter but not of the former cells. PMID- 2990640 TI - Vagal body tumor: paraganglioma of the head and neck. AB - Vagal body tumors (VBTs) are rare tumors of the paraganglion cells of the vagus nerve, usually occurring near the ganglion nodosum. They can be familial, multicentric, malignant, and sometimes hormonally active. The most accurate diagnosis is made by angiography. Primary treatment is surgical excision. Radiation therapy may play a role in preoperative preparation or in palliation. The prognosis is good when the tumor is found early and completely resected. Lifelong follow-up of patients and their family members is recommended. PMID- 2990641 TI - Different actions of vitamin D2 and D3 on bone metabolism in patients treated with phenobarbitone/phenytoin. AB - In 22 epileptic outpatients treated for at least 1 year with phenobarbitone/phenytoin the local and total bone mass, together with serum and urinary indices of calcium metabolism, were measured before and during treatment with either vitamin D2 or D3, 4,000 IU daily for 24 weeks. The results showed a distinct difference in the action of the two vitamins on bone metabolism during anti-convulsant treatment. The bone mass increased during treatment with vitamin D2, whereas the vitamin D3-treated patients showed unchanged values of bone mass, but an increased excretion rate of calcium, probably caused by increased intestinal calcium absorption. The data demonstrate that vitamins D2 and D3 (or their metabolites) have quantitative different effects in patients treated with phenobarbitone/phenytoin. PMID- 2990642 TI - Characterization of human bone cells in culture. AB - Cultures of human bone cells were established, maintained, and characterized with respect to several metabolic parameters. These studies were undertaken with a view to using the bone culture system as a means of studying mechanisms of bone metabolism. The donor patients' ages ranged from 1 to 90 years and their disease states included congenital limb anomalies, exostosis, and osteo- and rheumatoid arthritis. Cultures were maintained up to 5 months. The osteoblast-like character of these cells was confirmed with the use of measurements applied to bone cells from other systems. Analyses showed that (a) the cells' appearance resembled that of cultured osteoblasts from other animal sources, b) intracellular cAMP was stimulated by human parathyroid hormone, c) osteocalcin was detected in the medium of all tested bone cell cultures and its production was found to be stimulated by 1,25-dihydroxycholecalciferol, and d) newly synthesized collagen was almost exclusively type I. In contrast, cultures of human fibroblasts, established in one instance from tissue specimens of the same donor patient, grew faster, reached a higher limiting density, and produced a greater proportion of type III collagen than the corresponding bone cells. Furthermore, fibroblasts did not accumulate osteocalcin in their culture medium. The conditions described in this report to maintain human bone cells in culture should provide a suitable test system to study the regulation of human bone metabolism. PMID- 2990643 TI - Demonstration and characterization of 1,25-dihydroxyvitamin D3 receptors in basal cells of epidermis of neonatal and adult mice. AB - Nuclear and cytosolic receptors for 1,25-dihydroxycholecalciferol [1,25(OH)2D3] were demonstrated in the epidermis of neonatal and adult mice. The macromolecular binding protein sedimented at 3.5 S (sucrose density gradient) and was distinct from the 6.0 S binding protein for 25-hydroxycholecalciferol [25(OH)D3]. Analysis at different ionic strengths suggested the presence of unoccupied nuclear receptors. Digestion with proteases or nucleases, respectively, and inactivation with alkylating agents demonstrated that the binding macromolecule is a protein with SH groups at the active site. Binding of 1,25(OH)2D3 was specific and reversible. In neonatal mice KD was 1.6 X 10(-10) M for both cytosolic and nuclear fractions, binding capacity was 54 fmol/mg protein in the cytosolic and 108 in the nuclear fractions, respectively. The phenotypic expression of the 1,25(OH)2D3 receptor (dissociation constant, binding capacity) was identical in neonatal and adult epidermis. Half maximal displacement of 1,25(OH)2D3 was achieved with an 80-fold and 200-fold molar excess of 25(OH)D3 and 1-alpha hydroxycholecalciferol [1(OH)D3], respectively. Using Percoll density gradient centrifugation, 1,25(OH)2D3 receptors could be localized in the basal cell fraction. DNA cellulose chromatography with 1,25(OH)2D3 receptor elution from DNA at 0.25 M KCl (linear gradient) points to a possible role in gene transcription. In mouse primary epidermal cell cultures, 1,25(OH)2D3, but not 25(OH)D3, 24,25(OH)2D3, and 1(OH)D3 influenced [3H]thymidine incorporation (at physiological concentrations); the magnitude of change depending on the concentration of 1,25(OH)2D3 and the time of incubation. These data demonstrate that skin is a target organ for the active vitamin D secosterol. PMID- 2990644 TI - In vitro recombination of bacteriophage T7 DNA: further characterization of the reaction using plasmid DNA. AB - We have used a plasmid which contains a cloned fragment of T7 DNA to study the properties of general recombination of phage T7 in vitro. It was shown that T7 infected cell extracts promote recombination by the exchange of double strands of DNA. While both products of these double-strand exchanges were detected, we were unable to show that they were formed during a single recombination event. PMID- 2990645 TI - Phosphatidylcholine and 4-methylumbelliferyl phosphorylcholine hydrolysis by purified placental sphingomyelinase. AB - We present evidence which indicates that highly purified placental acid sphingomyelinase hydrolyses [14C]phosphatidylcholine [( 14C]PC) and the synthetic phosphodiester 4-methylumbelliferyl phosphorylcholine (4-MUPC). Hydrolysis was achieved by phospholipase C phosphodiesterase action. Of the several detergents tested, sodium taurocholate alone was necessary for PC hydrolysis, while 4-MUPC was hydrolysed independent of any detergent requirement. The pH optima for the reactions were 4.6-4.8 for PC hydrolysis and 4.8-5.0 for 4-MUPC hydrolysis. As with sphingomyelin hydrolysis, degradation of both PC and 4-MUPC was inhibited by 5'-, 3'-, and 2'-AMP, 5'-AMP being the most effective of the three. Furthermore, the phosphodiesterase activity against PC and 4-MUPC copurified with sphingomyelinase from human placenta and cross-reacted with a specific anti sphingomyelinase monoclonal antibody, strongly indicating identity of the phosphodiesterases. This explains phospholipase C deficiency in sphingomyelinase deficient Niemann-Pick disease cells. PMID- 2990647 TI - Site-specific cleavage of tobacco mosaic virus RNA: a study of factors influencing the cleavage. AB - DNA oligomer directed ribonuclease H (RNase H) methodology is applied to specifically cleave tobacco mosaic virus (TMV) RNA. Using a synthetic DNA oligomer P(dT8)dCdC, complementary to a region from nucleotide 5545 to nucleotide 5554 at the 3' end of TMV RNA, we have cleaved the RNA at the site of polynucleotides complementary to the DNA oligomer. Factors such as secondary structure of the RNA, concentrations of DNA oligomer, RNase H and magnesium ions in the reaction mixture, and time of incubation were optimized for the RNase H cleavage of TMV RNA-DNA oligomer complex. Denaturation of TMV RNA with 50% dimethyl sulphoxide at 50 degrees C is essential for the site-specific cleavage. PMID- 2990646 TI - Role of superoxide and hydrogen peroxide in cell lysis during irradiation in vitro of Ehrlich ascitic carcinoma cells in the presence of melanin. AB - The reactive species involved in the cell lysis during ultraviolet irradiation of Ehrlich ascitic carcinoma cells in the presence of red hair melanin (RHM) were investigated by determining 51Cr release from labeled cells. Cysteine at 1 mM in the presence of RHM increased the cell lysis during the incubation in the dark as well as during irradiation; this lysis was enhanced by superoxide dismutase (SOD). Catalase abolished the dark reaction and inhibited the cysteine-induced increase of cell lysis during irradiation. The cell lysis by the superoxide generating xanthine oxidase system was not significantly increased by SOD, but was significantly decreased by nitroblue tetrazolium and completely abolished by catalase. The cell lysis induced by the supernatants obtained from the suspensions of RHM either irradiated alone or with cysteine was abolished by catalase. Sediments of irradiated RHM when incubated in the dark with the cells did not release 51Cr. Irradiation of the cells in the presence of the same sediments produced lysis which was not inhibited by catalase. These studies suggest that superoxide per se is not toxic to the cells, but the H2O2 formed by dismutation of superoxide produces cell lysis either directly or by generating OH through Fenton-type reactions. A large part of the cell lysis seen during irradiation of cells in the presence of RHM is not due to H2O2, but may possibly be due to the melanin free radicals formed during irradiation. PMID- 2990648 TI - Pneumonia in calves produced with aerosols of Pasteurella multocida alone and in combination with bovine herpesvirus 1. AB - Pathological changes in respiratory tracts were studied in 30 calves following exposure to aerosols of Pasteurella multocida or to bovine herpesvirus 1 and P. multocida. Two groups of five calves were exposed to aerosols of one of two types of P. multocida only, which produced lobar pneumonia in one calf of each group. Another five groups of four calves were exposed to aerosols of bovine herpesvirus 1 and four to seven days later to one of the two types or one sub-type of P. multocida. Extensive necropurulent lesions were produced throughout the respiratory tract with each type of P. multocida in all four calves in three groups but none in the remaining two groups. The pathological changes differed from those produced following similar exposures to bovine herpesvirus 1 and P. haemolytica, in that the exudate in air passages and alveoli was more purulent and streaming (oat) cells and large mononuclear eosinophilic granulocytes were absent. This is the first report of experimental respiratory disease in cattle as a result of aerosol exposure to P. multocida alone or in combination with bovine herpesvirus 1. PMID- 2990649 TI - Sensitivity and specificity of an enzyme-linked immunosorbent assay for the detection of infectious bovine rhinotracheitis viral antibody in cattle. AB - An enzyme-linked immunosorbent assay was developed to detect bovine serum antibody to infectious bovine rhinotracheitis virus. The specificity of this assay in 304 bovine sera, collected from an infectious bovine rhinotracheitis virus-free herd, was 100%; in sera from 62 cattle inoculated with an intranasal vaccine, its diagnostic sensitivity was 27.4% at one month and 100% at six months, postvaccination. In 303 bovine sera with standard serum neutralizing antibody titers of greater than or equal to 1:2 it showed 100% sensitivity; and in 463 random diagnostic samples, comparative tests indicated that enzyme-linked immunosorbent assay detected more seropositive animals (61.6%) than the standard serum neutralizing test (49.9%). The enzyme-linked immunosorbent assay method was considered to be technically superior as a routine diagnostic test for the detection of infectious bovine rhinotracheitis viral antibody in bovine sera. PMID- 2990650 TI - Beware of breast fibroadenomas in middle-aged women. AB - On first examination of 8512 Quebec area women from 40 to 59 years of age who participated in the National Study of Breast Cancer Screening, the prevalence of fibroadenoma was 8.3 per 1000. In two women, a carcinoma was discovered within their fibroadenoma--intraductal carcinoma in one and an infiltrating ductal carcinoma in the other. Clinical and histopathologic findings are described and the prognosis and treatment discussed. The peak age of women at the time of diagnosis of a fibroadenoma is in their twenties while for those with a fibroadenoma containing a carcinoma, it is in the forties. For patients under 25 years, excision can be postponed for a few months since spontaneous regression may occur and the risk of breast cancer is small at that age. For older women, treatment should not be delayed. PMID- 2990651 TI - Laboratory evidence of viral and selected nonviral infections in humans in Canada, 1984. PMID- 2990653 TI - Dietary suppression of colonic cancer. Fiber or phytate? AB - The incidence of colonic cancer differs widely between various human populations. It has been suggested that dietary fiber content is of utmost importance and is inversely related to the occurrence of colonic cancer. However, high-fiber diets are not always correlated with low frequency of colonic cancer, suggesting the involvement of additional dietary constituents. Inositol hexaphosphate (phytic acid) is an abundant plant seed component present in many, but not all, fiber rich diets. The authors have found that phytic acid is a potent inhibitor of iron mediated generation of the hazardous oxidant, hydroxyl radical. Herein, the authors propose that inhibition of intracolonic hydroxyl radical generation, via the chelation of reactive iron by phytic acid, may help explain the suppression of colonic carcinogenesis and other inflammatory bowel diseases by diets rich in phytic acid. PMID- 2990652 TI - The Vancouver Lymphadenopathy-AIDS Study: 4. Effects of exposure factors, cofactors and HTLV-III seropositivity on number of helper T cells. AB - Results of testing for antibody to human T-lymphotropic virus (HTLV-III) and absolute numbers of helper T cells in 219 participants in the Vancouver Lymphadenopathy-AIDS (acquired immune deficiency syndrome) Study were analysed. The mean absolute helper T-cell counts in the 141 HTLV-III seronegative and the 78 seropositive men were 897/mL and 659/mL respectively (p less than 0.001). Established AIDS risk factors such as elevated lifetime number of male sexual partners and frequent receptive anal intercourse did not appear to have any significant effect on number of helper T cells that was independent of HTLV-III antibody status. Seropositive men with less than 100, 100 to 500 or more than 500 male sexual partners in their lifetime had mean absolute helper T-cell counts of 667/mL, 651/mL and 662/mL respectively. Most other risk factors, as well, did not appear to exert any effect on absolute number of helper T cells that was independent of the effect of HTLV-III antibody status. However, independent effects of a history of mononucleosis or hepatitis and of cigarette smoking were noted. The data support the hypothesis that no immune dysfunction beyond that due to the initial infection alone arises from repeated exposure to HTLV-III. Most risk factors appear to act as exposure factors, exerting their effect on the immune system merely by increasing the probability of contact with the agent. The independent effects of a history of mononucleosis or hepatitis suggest that viral agents may be cofactors in the production of immune dysfunction. PMID- 2990654 TI - Hypercalcemia in cats with feline-leukemia-virus-associated leukemia-lymphoma. AB - Three cases of hypercalcemia were recognized among 11 cats presenting with leukemia-lymphoma for ex vivo immunoadsorption therapy using Staphylococcal Protein-A-coated filters. In addition, the initial mean serum calcium concentration of cats with leukemia-lymphoma was significantly higher (P less than 0.005) than that of healthy control cats or feline-leukemia-virus-infected cats without malignancy. During immunotherapy of the hypercalcemic cats, objective reduction in the extent of the malignancies was associated with a small reduction in the serum calcium concentrations. This response to treatment, the lack of skeletal metastasis, and the absence of renal and parathyroid pathologic findings imply that humorally mediated mechanisms may have been responsible for the production of the hypercalcemia. PMID- 2990655 TI - Soft tissue sarcomas of the adult head and neck. AB - From 1969 to 1983, 53 adult patients with head and neck soft tissue sarcomas were evaluated and treated by the Division of Surgical Oncology at the University of Illinois. The most common anatomic location was the neck (36%), and these patients had the highest 5-year disease-free survival rate (67%). Fibrosarcoma was the most common histologic type (26%); patients with aggressive fibromatosis had the longest mean survival time (93 months). The mean overall survival time was 58.7 months, and the disease-free 2-year, 5-year, and 10-year survival rates were 68%, 54%, and 28%, respectively. Wide excision was the treatment of choice, with adjuvant radiotherapy or chemotherapy, or both, used in selected patients. In all of the long-term survivors, the tumors were either well-differentiated or less than or equal to 5.0 cm in diameter. It is apparent that aggressive therapy of such tumors can provide good long-term results. PMID- 2990656 TI - Serum copper level in ovarian carcinoma. AB - Serum copper levels (SCL) were determined before any diagnostic procedure was performed or treatment given in 40 women admitted for the investigation of a pelvic mass and, later, in those patients with ovarian carcinoma after chemotherapy and before a second-look operation was performed. Patients with ovarian carcinoma were found to have significantly higher SCL than patients with benign ovarian lesions. A SCL of 150 micrograms/dl clearly separated patients with a pelvic mass on the basis of ovarian carcinoma and those with benign noninflammatory pelvic lesions. The same SCL of 150 micrograms/dl separated patients with ovarian carcinoma that responded to chemotherapy and those with residual disease. It is suggested that SCL be included as a member of the screening panel of biologic tumor markers in general and in ovarian carcinoma in particular. PMID- 2990658 TI - The glioblastoma multiforme in Georgia, 1977-1981. AB - To characterize the incidence and mortality of the glioblastoma multiforme as it is seen at the primary care level, a retrospective study was done through the Georgia Tumor Registry on 299 patients with histologically confirmed supratentorial glioblastoma multiforme. Specifically, the age incidence distribution, sex ratio, and the postoperative survivals were studied. The Registry also provided the opportunity to define the relative racial frequencies of this neoplasm in a large biracial population. The mean age of the patients at the time of surgery was 58.6 years and the age group having the highest incidence was 60 to 74 years. The overall ratio of affected males to females was 1.04 to 1. The postoperative survival periods were considerably shorter than those reported from large cooperative clinical trials. The overall postoperative survivals at 12 and 24 months were only 22% and 4%, respectively. Advancing age had a strong negative relationship with the length of survival such that the 50% survival for patients less than 30 years of age was 17 months, for those age 30 to 44 years it was 10.2 months, for those age 45 to 59 it was 5.8 months, and for those older than 60 it was only 2.2 months. The study confirmed previous suggestions that the incidence of the neoplasm is considerably higher in whites. There were 2.3 glioblastomas in whites for every one in blacks. No difference was noted in the length of survival between whites and blacks. PMID- 2990657 TI - Teratoma with malignant transformation in germ cell tumors in men. AB - Pathology reports of over 580 male patients with germ cell tumors treated between 1972 and 1982 were screened for teratomas in which malignant transformation was apparent. The diagnosis was established in 17 cases. The median age was 28 years (range, 14-52). For patients with disease limited to the testis, the median survival has not been reached, with all five patients surviving disease-free at 22+ to 120+ months. Among the 12 patients with metastatic disease, the median survival was 30.5 months (range, 12-69). All 12 patients were treated with cisplatin-containing regimens. Six had complete responses either to chemotherapy alone or to chemotherapy plus resection of residual disease. Four of the six complete responders relapsed, and three died of progressive disease. One patient with sarcomatous differentiation was treated with a doxorubicin-based regimen and had a partial remission, which lasted 8 months. Teratoma with malignant transformation, when found in metastatic sites, appeared to be a poor prognostic pathologic variant in male patients with germ cell tumors. PMID- 2990659 TI - Mondor's disease associated with metastatic axillary nodes. AB - Mondor's disease, superficial thrombophlebitis of the breast, is customarily associated with benign conditions of the breast. This article reports a patient in whom an early manifestation of recurrent axillary metastasis from carcinoma of the breast was a symptom of ipsilateral superficial thrombophlebitis of the breast, an unusual association. PMID- 2990660 TI - Occupation and lung cancer risk. An analysis by histologic subtypes. AB - The relationships between cigarette smoking, occupational exposures, dietary vitamin A and lung cancer is explored in a case-control study. The study is based on 1002 primary lung cancers among white men and 1119 white male controls between 35 and 79 years of age admitted to Roswell Park Memorial Institute from 1957 to 1965. Four hundred sixty-four lung cancer cases were classified as one of the three major histologic cell types: squamous, small cell, or adenocarcinoma. Each of the three main histologic types displayed a statistically significant trend of increasing risk associated with increasing consumption of cigarettes with the strongest relationship being seen for small cell carcinoma (odds ratio for 40+ pack-years = 17.5) and the weakest with adenocarcinoma (odds ratio for 40+ pack years = 6.7). Increasing risk for squamous cell cancer was seen with increasing duration of exposure to all lung carcinogens, asbestos, and aromatic hydrocarbons. Small cell carcinomas were not associated with any of the occupational exposure categories studied with the exception of asbestos exposure in light smokers. Increasing risk of adenocarcinoma was seen with increasing duration of employment in occupations with potential exposure, but only for light smokers. Dietary vitamin A consumption did not confound the occupational associations. Hypotheses have been generated regarding interplay of risk factors in the etiology of different histologic subtypes of lung cancer. Studies of specific histologic types of lung cancer are needed to elucidate the role of various environmental and occupational risk factors. PMID- 2990661 TI - Natural history of hepatocellular carcinoma and prognosis in relation to treatment. Study of 850 patients. AB - A total of 850 patients with hepatocellular carcinoma seen during the last 8 years were analyzed retrospectively for survival in relation to treatment and disease stage. A new staging scheme based on tumor size, ascites, jaundice and serum albumin was used. Clearly, the prognosis depended on disease stage. The median survival of 229 patients who received no specific treatment was 1.6 months, 0.7 month for Stage III patients, 2.0 months for Stage II, and 8.3 months for Stage I. The median survival of Stage I patients who had hepatic resection (n = 115) was 25.6 months and Stage II patients with resection (n = 42) was 12.2 months. In patients who had a small cancer (less than or equal to 25% of liver area in size) the median survival was 29.0 months. Survival of the surgically treated patients, which represented a highly selected group, was better than that of medically treated patients of a comparable stage. Median survival of Stage I medically treated patients (n = 124) was 9.4 months, for Stage II (n = 290) 3.5 months, and for Stage III (n = 50) 1.6 months. Medical treatment prolonged survival in Stage II and III patients, but not in Stage I. Transcatheter arterial embolization gave a better survival compared with chemotherapy, whether intra arterial bolus administration of mitomycin C, systemic mitomycin C, or oral/rectal tegafur, in Stage II. Among various chemotherapeutic modalities, intra-arterial bolus injection was superior to systemic chemotherapy in survival in Stage II. In Stage III, chemotherapy improved survival as compared with no specific treatment. The major causes of death were hepatic failure and gastrointestinal bleeding, probably due to the coexistent advanced cirrhosis. These results in survival are much improved over the past reports, and the differences are probably a result of earlier diagnosis and frequent hepatic resections. PMID- 2990662 TI - Clinical, serologic, and immunologic effects of human leukocyte interferon in HBsAg-positive primary hepatocellular carcinoma. AB - Five patients with hepatitis B surface antigen (HBsAg) positive chronic hepatitis and histologically confirmed primary hepatocellular carcinoma (PHC) were treated with 3 X 10(6) units/day of partially purified human leukocyte interferon intramuscularly for 2 consecutive months. During interferon therapy, one patient had stable disease, while the remaining four patients had progressive disease. Following interferon therapy no changes were noted in the hepatitis B viral markers or in serum alphafetoprotein levels. Data on the effects of human leukocyte interferon on lymphocyte subpopulations and on the cytotoxic activity of peripheral blood mononuclear cells against a hepatitis B surface antigen expressing primary hepatocellular carcinoma cell line are presented. PMID- 2990663 TI - Chemotherapy for metastatic Merkel cell carcinoma. AB - Merkel cell carcinomas (also known as trabecular carcinomas) are primary cutaneous small cell neuroendocrine neoplasms with the potential to metastasize. Control of disseminated disease is therefore important. A case of metastatic Merkel cell carcinoma with an excellent response to chemotherapy is presented. The regimen chosen for this case is similar to that used for bronchogenic small cell anaplastic carcinoma. The reason for selecting this regimen was the common neuroendocrine differentiation and the similar histopathologic features of these two tumors. Only a few reports have described chemotherapy for Merkel cell carcinoma and similar agents were used. These cases are reviewed and critically analyzed. PMID- 2990665 TI - Anticancer drug screening test with LDH in nude mouse bearing bone and soft part sarcoma. AB - With the use of human LDH as a marker, growth and remission of four human osteosarcomas (KOS-1, KOS-2, KOS-3, OST) and a malignant fibrous histiocytoma (KMF) transplanted into nude mice were monitored during chemotherapies (Adriamycin [doxorubicin], cisplatin, mitomycin C, cyclophosphamide and vincristine). Four osteosarcomas were effectively treated with Adriamycin and cisplatin, and KMF was treated with Adriamycin, cisplatin, and mitomycin C. Changes in human LDH levels paralleled changes in tumor size and these could be used to predict the death of the animals due to tumor (P less than 0.01). Furthermore, the enzyme increased transiently after administration of the effective drugs. PMID- 2990664 TI - Glioblastoma multiforme and anaplastic astrocytoma. Pathologic criteria and prognostic implications. AB - A total of 1440 malignant astrocytic gliomas from three Phase III trials of the National Brain Tumor Study Group were studied to document the clinical usefulness of subclassifying these lesions as either an anaplastic astrocytoma or a glioblastoma multiforme. As defined by a previous "blind" pathology review, the two groups of patients were compared as to mean age, mean duration of preoperative symptoms, and postrandomization survival. In addition, 10 histologic variables were studied in 150 patients with the anaplastic astrocytoma to establish internal correlations, and to relate specific histologic variables to patient age and postrandomization survival. There were highly significant differences in the age, duration of preoperative symptoms, and post randomization survival between the two groups. Internal correlations between histologic variables in the anaplastic astrocytoma disclosed statistically significant associations between the presence of lymphocytes and gemistocytic astrocytes. It is concluded that the subclassification of malignant gliomas into the anaplastic astrocytoma and the glioblastoma multiforme defines groups of patients that are significantly different in regard to age, duration of symptoms, and length of survival. The problems of tissue sampling are recognized, however, the assignment, by a blind pathology review, to two such different groups indicates that the classification has utility for large randomized clinical trials. The analysis of histologic variables in the anaplastic astrocytomas confirms previous suggestions that lymphocytes and gemistocytes frequently coexist in malignant gliomas, but in this study these inflammatory cells did not appear to influence survival. The study reemphasizes the association between advancing age and shorter survivals in patients with malignant gliomas. PMID- 2990666 TI - Race-related morphologic variations in hepatocellular carcinoma. AB - In West Malaysia, hepatocellular carcinoma (HCC) is common in the Chinese and in the members of the Senoi aboriginal tribe, two racial groups with diametrically opposite life-styles. Certain fundamental differences exist between the liver tumors in the two races. In the Senoi, the tumor occurs in a younger age group and there is a greater male preponderance than in the Chinese. There is also a very close relationship between hepatitis B virus infection, chronic active hepatitis, cirrhosis, liver cell dysplasia, and HCC in the Senoi and the tumors generally present as multiple nodules studding both lobes of the liver. In the Chinese, although a relationship between hepatitis B virus infection, HCC, and cirrhosis exists, this association is not as strong as in the Senoi and the tumors are generally large and solitary. The data suggest that, although the hepatitis B virus is probably an important oncogenic agent in both racial groups, there may be a difference in the pathogenesis of HCC in the two races. PMID- 2990667 TI - Adnexal carcinomas of the skin. I. Eccrine carcinomas. AB - The diagnostic identification of sweat gland carcinomas is hampered by their rarity and their histologic resemblance to various visceral tumors, leading to confusion with metastatic lesions. In this series, 14 cases of eccrine carcinoma in five male and nine female patients, ranging in age from 13 to 84 years, are described. Ten tumors strongly resembled infiltrating ductal adenocarcinomas of the breast, and were thus classified as ductal. Three had a prominent mucinous matrix, similarly explaining their categorization as mucinous carcinomas. Finally, one neoplasm was a classic eccrine porocarcinoma. Four patients with ductal eccrine carcinomas suffered metastasis, and a 50% mortality rate was observed among this group of ten cases. In contrast, only one of three mucinous carcinomas metastasized, although all of these lesions recurred locally, as did the single porocarcinoma. None of the latter four neoplasms proved fatal. The results of conventional special stains in these 14 cases are discussed, and histologic features that they shared, and which may be utilized in distinguishing eccrine carcinomas from benign sweat gland tumors, are presented. PMID- 2990668 TI - Histologic multifocality of Tis, T1-2 breast carcinomas. Implications for clinical trials of breast-conserving surgery. AB - Breast cancer multifocality was studied in mastectomy specimens by correlated specimen radiography and histologic techniques. The patients chosen for study were comparable to those eligible for breast-conserving surgical therapy. Two study groups, one with 282 invasive cancers (T1-2) and the other with 32 intraductal cancers, were selected from a group of 399 consecutive cases by omitting patients who were clearly, or very probably, not candidates for breast conserving surgical therapy according to current trial criteria. Omitted patients included those with clinically and/or radiologically multifocal cancers and patients with tumor extension into the chest wall or skin (7%). Also excluded were the so-called diffuse invasive cancers (8%), the clinically and radiologically occult tumors (3%), and the invasive cancers larger than 5 cm (3%). Of the 282 invasive cancers, 105 (37%) showed no tumor foci in the mastectomy specimen around the reference mass. In 56 (20%) tumor foci were present within 2 cm, and in 121 (43%) tumor was found more than 2 cm from the reference tumor. In 75 (27%) the tumor foci beyond 2 cm were histologically noninvasive cancers, and in 46 cases (16%) they contained invasive cancers as well. A comparison between the group with reference tumors less than 2 cm and the group with reference tumors more than 2 cm in size showed no significant difference between the groups in terms of presence or absence of tumor foci or distance of tumor foci from the reference tumor. If the 264 invasive cancers in this series that were 4 cm or less in diameter had been removed with a margin of 3 to 4 cm, 7% to 9% of the patients would have had invasive cancer left in the remaining breast tissue, and 4% to 9% would have had foci of noninvasive cancer left in the remaining breast tissue. On the basis of the data on the distribution of tumor at different distances from the reference tumor, the current study estimates the expected rates of local recurrences after breast-conserving surgical procedures relative to the extensiveness of the excision. The possible impact of postoperative local radiation therapy on the rates of expected local recurrence is discussed. PMID- 2990669 TI - Induction of nitroblue tetrazolium reduction in mouse peritoneal macrophages by tumour promoters and inhibition of the induced nitroblue tetrazolium reduction by some inhibitors. AB - Two polyacetates, aplysiatoxin and debromoaplysiatoxin, as well as 12-O tetradecanoylphorbol-13-acetate (TPA), mezerein and teleocidin enhance nitroblue tetrazolium (NBT) reduction in mouse peritoneal macrophages in vitro. The ED50 values for NBT reduction of these 5 TPA-type tumor promoters were 4.2 ng/ml for TPA, 36 ng/ml for mezerein, 0.53 ng/ml for teleocidin, 1.5 ng/ml for aplysiatoxin and 108 ng/ml for debromoaplysiatoxin. The NBT reduction induced by the 5 tumor promoters is inhibited by 2 inhibitors of tumor promotion, retinoic acid and dibromoacetophenone. The possibility that tumor promotion by TPA-type tumor promoters involves similar mechanisms such as superoxide anion radicals release in cell membranes is discussed. PMID- 2990670 TI - The subpopulations and isolated cell types of freshly resected high grade human gliomas: their influence on the tumor's evolution in vivo and behavior and therapy in vitro. AB - Human malignant gliomas are karyotypically heterogeneous, composed of many cellular populations and isolated cell types identifiable by cytogenetic techniques. The distributions of cell types vary in high grade tumors. Some tumors are primarily near-diploid (35-57 chromosomes per cell), while others are hyperdiploid with chromosome numbers ranging from 58 to several hundred chromosomes per cell. Regional studies of several tumors suggest that the heterogeneity is not random. Anatomically different regions result from the combination of cellular distribution and their evolution over time. The karyotypic pattern of gliomas also reflects the tumor's evolution from a relatively homogeneous population of near-diploid cells to the hyperdiploid tumor that is removed by the neurosurgeon. The in vitro studies also suggest that there are phenotypic correlates to the karyotypic pattern of the tumor cells. Hyperdiploid cells are unstable in culture, tend to grow rapidly with short doubling times, and are often sensitive to such chemotherapeutic agents as BCNU. In contrast, the near-diploid cells are more normal in appearance, are stable in culture, grow slowly with long doubling times, are more likely to be resistant to the nitrosoureas and ultimately are the 'stem' cells that repopulate the tumor mass. PMID- 2990671 TI - Human T-cell leukemia virus I induction by 5-iodo-2'-deoxyuridine and N-methyl-N' nitro-N-nitrosoguanidine: inhibition by retinoids, L-ascorbic acid, and DL-alpha tocopherol. AB - Human T-cell leukemia virus type I was induced by N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and 5-iodo-2'-deoxyuridine (ldUrd) in the MT-1 cell line. Virus expression was monitored by immunofluorescence microscopy with GIN-14, mouse monoclonal antibodies directed toward Mr 19,000 and Mr 28,000 protein specific virus polypeptides. MNNG (0.1 micrograms/ml) and ldUrd (50 micrograms/ml) both induced virus synthesis in MT-1 cells. MNNG-induced virus expression peaked between 24 and 48 h of incubation, whereas ldUrd induced maximum virus expression between 48 and 72 h of incubation. Superinduction resulted when MNNG was added to cells induced 48 h previously with ldUrd, but not with concomitant treatment. 13-cis-Retinoic acid, retinol, retinol aldehyde, and retinol acetate (10(-6) to 10(-9)M) were concomitantly added with ldUrd to MT-1 cells for 24, 48, and 72 h incubation. All inhibited virus induction to various degrees. The retinoids were ranked as to inhibitory activity: retinol greater than retinoic acid greater than retinol aldehyde greater than retinol acetate. The most sensitive period for inhibiting ldUrd induction by retinoic acid was 24 h postinduction or with concomitant treatment. Vitamin C and vitamin E inhibited ldUrd induction most effectively with 48 h incubation. Retinol and vitamin C also inhibited virus induction by MNNG. None of the retinoids, vitamin C, or vitamin E significantly inhibited virus expression in noninduced cells or were toxic to the cells at the concentrations used in these experiments. PMID- 2990672 TI - Hydrogen peroxide as a tumoricidal mediator of murine polymorphonuclear leukocytes induced by a linear beta-1,3-D-glucan and some other immunomodulators. AB - Polymorphonuclear leukocytes (PMN) of mice can destroy tumor cells effectively in vitro in the presence of antitumor polysaccharide, linear beta-1, 3-D-glucan from Alcaligenes faecalis var. myxogenes IFO 13140 (TAK), and some other immunomodulators. In the present study, we investigated the mechanism of the tumoricidal activity of PMN induced by these immunomodulators and especially TAK. The TAK-induced PMN cytotoxicity was concluded to involve hydrogen peroxide from the following results: (a) the cytotoxicity depended on glucose consumption; (b) it was almost completely inhibited by catalase but not affected by superoxide dismutase; (c) it was not reduced by cyanide or azide, which are inhibitors of myeloperoxidase; (d) it was not affected by scavengers of singlet oxygen or hydroxyl radical; (e) release of hydrogen peroxide from PMN was observed by the addition of TAK; (f) MM46 target cells were lysed directly by hydrogen peroxide in the absence of myeloperoxidase; (g) the supernatant of PMN in the presence of TAK, tested as a stable cytotoxic factor, did not have cytotoxic activity, and protease inhibitors had no effect on this cytotoxicity. These results suggest that hydrogen peroxide is a direct cytotoxic mediator in TAK-induced PMN cytotoxicity. Next, the mechanism of PMN cytotoxicities induced by other immunomodulators was also examined and was compared with that induced by TAK. The results suggest that hydrogen peroxide is also important for these cytotoxicities whereas, unlike the results with TAK, the H2O2:halide:myeloperoxidase system may partly participate in the cytotoxicities with some immunomodulators. PMID- 2990673 TI - Transferrin-like activity produced by murine malignant T-lymphoma cell lines. AB - Transferrin has been considered to be an essential requirement for hematopoietic cell proliferation in culture. We have isolated two cloned lymphoma cell lines, SL 12.1 and SL 12.4, which grow and adapt in serum-free medium without added transferrin. Antibody to the transferrin receptor blocks the growth of these cells. We have also demonstrated that transferrin-free conditioned medium from the cells will compete with transferrin for binding. Furthermore, conditioned medium from SL 12.1 and SL 12.4 cells induces and supports exponential growth of a transferrin-dependent lymphoma cell line, SL 12. We conclude that these two transferrin-independent cloned lines produce transferrin-like activity which plays a crucial role for cell proliferation. PMID- 2990674 TI - Radiosensitization effects of nicotinamide on malignant and normal mouse tissue. AB - Inhibitors of the chromatin-associated enzyme adenosine diphosphate ribosyltransferase have been found to inhibit DNA strand rejoining and to potentiate lethality of DNA-damaging agents both in vivo and in vitro. We have in this work examined the radiosensitizing potential of one such inhibitor, nicotinamide, on tumor tissue by using transplanted C3H mouse mammary adenocarcinomas and on normal tissue in a tail-stunting experiment using BALB/cA mice. Our data indicate a radiosensitizing effect of nicotinamide on tumor cells as well as on normal tissue. The data indicate a possible role of adenosine diphosphate ribosyltransferase inhibitors as a sensitizing agent in the radiotherapy of malignant tumors. PMID- 2990676 TI - Alterations in the expression of a hepatocyte cell adhesion molecule by transplantable rat hepatocellular carcinomas. AB - Alterations in the expression of normal cell surface components on 13 transplantable hepatocellular carcinomas were examined using a heteroantiserum [anti-Mr 105,000 glycoprotein (gp 105)] reactive with a family of nine wheat germ agglutinin binding components from normal rat hepatocytes with an average molecular weight of 105,000. Analysis by two-dimensional polyacrylamide gel electrophoresis of components immunoprecipitated by anti-gp105 antiserum from detergent extracts of transplantable hepatocellular carcinoma cells surface labeled with 125I revealed qualitative and quantitative changes in the expression of anti-gp105-reactive components with the most consistent change being the apparent loss of a pair of acidic (pl 4.1 to 4.3) glycoproteins by all 13 transplantable hepatocellular carcinoma lines. One-dimensional peptide maps of fragments produced following digestion with V8 protease indicated that these acidic components were closely related in structure but differed significantly from other anti-gp105-reactive components. Immunodepletion analysis with monoclonal antibodies and heteroantisera reactive with individual components recognized by anti-gp105 antiserum showed that the two acidic glycoproteins were antigenically and structurally identical to cell-CAM 105, a Mr 105,000 glycoprotein involved in cell-cell adhesion of rat hepatocytes. Antibodies raised against purified cell-CAM 105 were specific in immunoprecipitation assays for the acidic components, strongly inhibited reaggregation of hepatocytes, and displayed no reactivity by indirect immunofluorescence or immunoprecipitation analysis with transplantable hepatocellular carcinoma cells. These results suggest that major alterations in the expression of cell-CAM 105 may be a consistent feature of the malignant phenotype. PMID- 2990677 TI - Stability of vincristine complexes in cytosols derived from xenografts of human rhabdomyosarcoma and normal tissues of the mouse. AB - The selective action of vincristine (VCR) has been correlated with longer retention of the drug in neoplastic tissue compared with normal tissues of the mouse (J. A. Houghton, L. G. Williams, P. M. Torrance, and P. J. Houghton, Cancer Res., 44: 582-590, 1984). In order to examine the basis for this differential, the stability of drug-protein complexes was examined further. The stability of drug-protein complexes formed in cytosols derived from HxRh18 tumors, ileum, liver, kidney, skeletal muscle, blood, brain, spleen, lung, and bone marrow was examined. Protein-bound [3H]VCR was isolated by gel filtration of [3H]VCR-cytosol mixtures from each tissue except for ileum and blood. Complexes formed in brain and HxRh18 cytosols were stable at 37 degrees for at least 2 h; all other complexes were unstable. For liver, kidney, and muscle, half-times of complexes were in a similar order to the initial rates of elimination of [3H]VCR from these tissues in vivo but were of shorter duration. The HxRh18-[3H]-VCR complex was unstable at 37 degrees in the presence of cytosols prepared from ileum, kidney, liver, and lung. Drug metabolism by these tissues was not detected in vitro. In the presence of heat-treated extracts from ileum or kidney, [3H]VCR complex was stable, suggesting that the destabilizing factor may be enzymic. Degradation of 125iodinated tubulin, analyzed by polyacrylamide-sodium dodecyl sulfate gel electrophoresis, occurred in the presence of ileum but not skeletal muscle or brain cytosols. This correlated with the stability of HxRh18-[3H]VCR complexes. In the presence of kidney cytosol, however, the molecular weight of 125I-tubulin remained unchanged, suggesting a different mechanism. Based upon data obtained, cytosols from normal tissues may be categorized into three classes: (a) those that formed stable complexes (brain); (b) those that formed unstable complexes but also destabilized preformed complex (ileum, kidney, liver, lung); and (c) tissues that formed unstable complexes but did not destabilized preformed complex (skeletal muscle, spleen, bone marrow, blood). The degree of instability of complexes formed in cytosols prepared from normal tissues appears to correlate with rapid loss of VCR from these tissues in vivo and hence may represent mechanism(s) for the selective action of this antineoplastic agent. PMID- 2990675 TI - Changes in cyclic adenosine 3':5'-monophosphate-dependent protein kinases during the progression of urethan-induced mouse lung tumors. AB - The cyclic adenosine 3':5'-monophosphate (cAMP)-dependent protein kinases in lung adenomas are functionally different from those of normal lung. The relevance of this change to neoplastic conversion was examined by comparing tumor kinases with those obtained from the normal cell of origin and by studying the kinases at different stages of tumor growth. Lung tumors were collected from A strain mice at different times after a single injection of urethan. These tumors are predominantly of alveolar type two cell origin, and cAMP-binding proteins in extracts from isolated type two cells and from lung adenomas at various stages of tumor progression were compared. Both the incorporation of the cAMP photoaffinity analogue, cyclic 8-azidoadenosine 3':5'-[32P]monophosphate (8-N3-[32P]cAMP), into the regulatory subunits of the type I (RI) and type II (RII) cAMP-dependent protein kinases and the autophosphorylation of RII were similar in extracts from whole normal lung and from type two cells. Altered protein kinases are thus not characteristic of normal type two cells. Lung tumors showed a decrease in photodetectable RII which correlated in degree with tumor size and extent of anaplasticity. This decreased RII photolabeling during tumor growth was associated with increased RII autophosphorylation. In contrast, decreased RII photolabeling in extracts from neonatal lung is accompanied by a substantial decrease in RII autophosphorylation. The characteristics of RII during normal development thus clearly differ from those during neoplastic development. An increase in the amount of an Mr 37,000 proteolytic fragment derived from R subunits was also noted as a function of tumor progression. DEAE-cellulose chromatography of tumor cytosol showed that the increase in the amount of Mr 37,000 protein was accompanied by increased subunit dissociation of the type I isozyme. The dissociated RI subunit has been shown to be more sensitive to cleavage by a Ca2+-dependent neutral protease than when RI was in the holoenzyme form. This protease is present in both normal lung and lung adenomas, and its activity increases during the later stages of tumor progression. A comparison of cAMP binding and the light-induced covalent incorporation of 8-N3-[32P]cAMP showed that, for both RI and RII, photoincorporation was about 75% as efficient as noncovalent binding. In contrast, although the Mr 37,000 fragment can be photolabeled with low concentrations of 8-N3-[32P]cAMP, noncovalent cAMP binding to the endogenous Mr 37,000 fragment could not be demonstrated with a standard filtration assay. Such altered cAMP binding characteristics following Ca2+ dependent proteolysis of R-subunits would all PMID- 2990678 TI - Effects of adrenocorticotropic hormone, human chorionic gonadotropin, and insulin on steroid production by human adrenocortical carcinoma cells in culture. AB - Adrenocortical carcinoma tissue removed from a mildly hirsute 16-year-old girl was cultured in order to assess steroidogenesis and responsiveness of the cells to adrenocorticotropic hormone (ACTH), human chorionic gonadotropin (HCG), and insulin. The cells in culture produced large amounts of androstenedione and testosterone; however, production of cortisol, which was initially high, decreased with time. No aldosterone, estrone, or estradiol was produced in vitro. Both monolayer cells maintained for 6 weeks and organ culture explants maintained for over 3 days responded to ACTH (10(-7) M) with increased production of androgens (testosterone, androstenedione, dehydroepiandrosterone) but decreased production of cortisol as measured by radioimmunoassay of steroids in the culture media. Concomitant with decreased cortisol production was the enhanced formation of 11-deoxycortisol in cells exposed to ACTH, suggesting impaired 11 beta hydroxylation. Tissue exposed to HCG (10(-7) M) in organ culture showed an increase in androgen production over control levels, but no significant effect of HCG on glucocorticoid production was found. Tumor cells differed in their androgen response to ACTH and HCG, with enhanced adrenal androgens in the presence of ACTH and more gonadal-type androgens after exposure to HCG. Insulin exposure had no effect on production of either androgen or glucocorticoid by tumor tissue in organ culture. Thus, this adrenocortical carcinoma showed marked androgen production in culture which was enhanced in different ways by ACTH and HCG. 11 beta-Hydroxylation was impaired with time in culture. No specific effect of insulin on steroidogenesis was noted. PMID- 2990679 TI - First international symposium on Epstein-Barr virus and associated malignant diseases. PMID- 2990680 TI - Symposium on HTLV. Bethesda, Maryland, December 6 & 7, 1984. PMID- 2990681 TI - The human T-cell leukemia/lymphotropic retroviruses (HTLV) family: past, present, and future. PMID- 2990682 TI - Retroviruses associated with leukemia and ablative syndromes in animals and in human beings. AB - T-lymphotropic retroviruses of cats cause lymphopenia and immunosuppression and represent the major cause of death in that species. Similarly HTLV-I which is T4 tropic is associated with an increased risk for development of infectious disease in regions where the virus is endemic. Since HTLV-I is also believed to be transmitted by blood and by sexual intercourse we considered the possibility that a variant form of HTLV might cause AIDS. The identification of cross-reactive antibodies to HTLV-I-MA in a third or more of the AIDS patients and in suspicious blood donors that donated to transfusion-associated cases of AIDS eventually led to the recognition of HTLV-III, the causative agent of AIDS. The protein most associated with lymphocyte immortalization or transformation in the case of HTLV I is p42. The proteins of HTLV-I encoded by the amino terminus of the env gene designated gp61 and gp45 are the most immunogenic antigens of this virus. Similarly those encoded by the amino terminus of the env gene HTLV-III designated gp160 and gp120 appear to be the most immunogenic markers for this agent. Almost all AIDS patients, ARC patients, and asymptomatic hemophiliacs have detectable antibodies to gp120 and gp160. HTLV-III related agents designated STLV-III have been found in macaque monkeys that develop simian AIDS and high prevalence rates of antibodies to STLV-III can be found in healthy African green monkeys. We hypothesize that the STLV-III of African green monkeys could represent a recent source of the virus to have infected humans in central Africa where the human epidemic probably began. The recognition that up to one million people may already be infected with HTLV-III in the United States alone indicates the need for development of a vaccine. The availability of primate species infected with the serologically related STLV-III agents that either resist disease development (African green monkeys) or succumb to an AIDS-type syndrome (rhesus) provide models that should aid in our attempts to develop such vaccines. PMID- 2990683 TI - Molecular biology of human T-lymphotropic retroviruses. AB - The generic name for a family of human T-lymphotropic retroviruses is HTLV. Two of the three members in this family have been linked etiologically to human diseases: HTLV-I with adult T-cell leukemia and HTLV-III with the acquired immunodeficiency syndrome. In addition to their T-cell tropism and a number of other common biological and biochemical properties, the most unique common features of these viruses from a molecular biological point of view are the presence of the x-lor gene towards the 3' end of the genome and the phenomenon of a virus-induced trans-acting factor in activation of transcription initiated in the viral long terminal repeat. These features may not only be key in understanding the mechanism of transformation or cell killing by these viruses, but they also provide a basis for new classification of retroviruses. In spite of these similarities among HTLV-I, -II, -III, and bovine leukemia virus, the genome of HTLV-III is only distantly related to these other viruses. Instead, it shows greater homology to members of the Lentivirus family. Therefore, all these viruses may have a common progenitor. Two other salient features arose from the analyses of HTLV-III and acquired immunodeficiency syndrome. (a) HTLV-III frequently infects the brain of acquired immunodeficiency syndrome patients who suffer from central nervous system disorders. This not only identifies HTLV-III as the direct candidate in these central nervous system disorders but also poses the problem of crossing the blood-brain barrier in therapy strategies to eradicate the virus. (b) Different HTLV-III isolates comprise a spectrum of related viruses, with the degree of divergence varying from virtual identity to 10-15% difference. The most divergent region resides in the envelope gene. Whether this finding has implications in the development of an effective vaccine for acquired immunodeficiency syndrome remains to be determined. PMID- 2990684 TI - The lor gene and pathogenesis of HTLV-I, -II, and -III. PMID- 2990686 TI - Heteroduplex mapping in the molecular analysis of the human T-cell leukemia (lymphotropic) viruses. AB - The human T-cell lymphotropic virus (HTLV) family includes members associated with T-cell cancers (HTLV-I and HTLV-II) as well as the etiological agent of the acquired immunodeficiency syndrome (HTLV-III). Molecular clones of these viruses were used in heteroduplex mapping experiments to study their structural and evolutionary relationships. The HTLV-I subgroup, despite some restriction enzyme site polymorphism, demonstrated a high degree of sequence conservation. Heteroduplexes of HTLV-I and HTLV-II demonstrated a significant amount of sequence homology, with the strongest region of conservation occurring in the 3' most coding sequences, designated pX, and to a lesser, although substantial extent in the rest of the genome. Thus, the genomic organization of HTLV-II appears to be very similar to that of HTLV-I. All HTLV-III molecular clones appeared to be identical, with a single exception, which showed heterogeneity in the env gene region. In heteroduplexes between HTLV-I and HTLV-III, very little homology was observed, being confined to the gap/pol region. In contrast to the latter result, a striking amount of homology was detected between HTLV-III and a morphologically similar, pathogenic, nononcogenic lentivirus, visna virus. These data provide strong evidence for a close taxonomic and thus evolutionary relationship between HTLV-III and the lentivirus subfamily of retroviruses. A taxonomic tree, based on the genetic relatedness and biological properties of the HTLV family, is proposed. PMID- 2990685 TI - Human T-cell leukemia virus x gene. AB - The human T-cell leukemia virus (HTLV) types I and II are associated with specific hematological cancers. These viruses rapidly transform normal T lymphocytes in vitro. The mechanism of HTLV-induced leukemogenesis is unknown. Structural analysis of HTLV-I and HTLV-II has revealed sequences of unknown function, termed X, at the 3' end of the proviral genome. The distal two-thirds of the X sequences are highly conserved between HTLV-I and HTLV-II. We have shown that these conserved X sequences contain a gene, termed x, that is expressed in both HTLV-I and HTLV-II by identifying a subgenomic X RNA as well as the proteins encoded by these messages. The function of this unique x gene is unknown; however, its conservation and expression suggest that it may play a role in HTLV replication and in HTLV-induced leukemogenesis. PMID- 2990687 TI - Interleukin 2 receptor (Tac antigen) expression in HTLV-I-associated adult T-cell leukemia. AB - Interleukin-2 (IL-2) is a lymphokine synthesized by some T-cells following activation. Resting T-cells do not express IL-2 receptors, but receptors are rapidly expressed on T-cells following interaction of antigens, mitogens, or monoclonal antibodies with the antigen-specific T-cell receptor complex. Using anti-Tac, a monoclonal antibody that recognizes the IL-2 receptor, the receptor has been purified and shown to be a Mr 33,000 peptide that is posttranslationally glycosylated to a Mr 55,000 mature form. Normal resting T-cells and most leukemic T-cell populations do not express IL-2 receptors; however, the leukemic cells of the 11 patients examined who had human T-cell lymphotropic virus-associated adult T-cell leukemia expressed the Tac antigen. In human T-cell lymphotropic virus-I infected cells, the Mr 42,000 long open reading frame protein encoded in part by the pX region of this virus may act as a transacting transcriptional activator that induces IL-2 receptor gene transcription, thus providing an explanation for the constant association of IL-2 receptor expression with adult T-cell lymphotropic virus-I infection of lymphoid cells. The constant expression of large numbers of IL-2 receptors which may be aberrant may play a role in the uncontrolled growth of adult T-cell leukemia cells. Two patients with Tac positive adult T-cell leukemia have been treated with the anti-Tac. One of the patients had 6- and 3-mo remissions of his leukemia following two courses of therapy with this monoclonal antibody directed toward this growth factor receptor. PMID- 2990688 TI - Isolation and expression of complementary DNAs encoding the human interleukin 2 receptor. AB - Complementary DNAs corresponding to the human receptor for interleukin 2 (IL-2) have been molecularly cloned, sequenced, and expressed in COS-1 cells. The human genome appears to contain a single structural gene for this receptor; however, when transcribed at least two messenger RNAs (mRNAs) are produced which vary in length due to the use of different polyadenylation signals. Sequence analysis of the cloned complementary DNAs indicates an alternate pathway of mRNA processing for this receptor. Splicing of a 216 base pairs segment contained within the protein coding region results in an mRNA unable to code for the IL-2 receptor. In contact complementary DNAs corresponding to unspliced mRNA encode membrane receptors which bind both IL-2 and anti-Tac (monoclonal anti-IL-2 receptor antibody). Analysis of the deduced amino acid sequence reveals that the receptor is composed of 272 amino acids including a signal peptide 21 amino acids in length. Hydrophobicity analysis suggests a single 19 amino acid transmembrane domain. A short intracytoplasmic domain composed of 13 amino acids is present at the carboxy terminus and contains three potential phosphate acceptor sites (serine and threonine but not tyrosine) and typical positively charged amino acids presumably involved in cytoplasmic anchoring. Two sites for N-linked glycosylation sites and numerous extracytoplasmic O-linked glycosylation sites are present. PMID- 2990689 TI - Pharmacological inhibition of infectivity of HTLV-III in vitro. AB - Acquired immunodeficiency syndrome (AIDS) is a pandemic immunosuppressive disease that predisposes to life-threatening opportunistic infections and unusual forms of neoplasms. A recently discovered member of the human T-lymphotrophic virus (HTLV) family, designated HTLV-III, has been shown to be the etiological agent of AIDS. We have shown previously that a trypanosomicidal drug, suramin, can block the in vitro infectivity and cytopathic effect of HTLV-III at doses that are attainable in human beings. In the present work we report our findings that suramin can block the cytopathic effect of HTLV-III even after a defined exposure of the target helper/inducer T-cells to the virus and that the T-cells protected by suramin remain immunologically functional. PMID- 2990690 TI - Advances in the isolation of HTLV-III from patients with AIDS and AIDS-related complex and from donors at risk. AB - During the last 2 yr more than 100 independent isolates of human T-cell leukemia virus type III have been obtained in this laboratory. Most isolates were from peripheral blood T-lymphocytes established in cell culture from acquired immunodeficiency syndrome (AIDS) and acquired immunodeficiency syndrome-related complex (ARC) patients and healthy donors at risk for AIDS. Several were also obtained from leukocytes from bone marrow, lymph node, and brain tissue and from body excretions, e.g., saliva and semen. In addition HTLV-III was found in cell free plasma. The incidence (number of isolates per number of patients or donors tested) of virus isolation was approximately 80% for ARC patients, approximately 50% for AIDS patients, and approximately 30% for healthy individuals at risk for AIDS. Inclusion of hydrocortisone in cell culture medium greatly facilitated the isolation of virus from primary leukocytes from AIDS/ARC patients and also promoted its transmission to fresh leukocytes in vitro. Biological analysis of cells from infected patients or donors, as well as from normal peripheral blood mononuclear cells exposed to virus in vitro, demonstrated that OKT4/leu3a+ T lymphocytes were preferentially infected and were subjected to a characteristic cytopathic effect. In addition to the well-defined individuals at risk for AIDS, heterosexual transmission of HTLV-III with its subsequent pathological manifestations was found. Virus was isolated from males with heterosexual promiscuity as their only recognized risk factor and from the spouses of these and other AIDS and ARC patients. PMID- 2990692 TI - Epidemiological trends of AIDS in the United States. AB - The incidence of acquired immunodeficiency syndrome (AIDS) in the United States has increased rapidly since the first reports in 1981. Highest estimated rates are among single (never-married) men in Manhattan and San Francisco, men and women who have abused drugs intravenously, and persons with hemophilia. Serosurveys among populations at increased risk for AIDS have demonstrated a high prevalence of antibody to human T-cell leukemia virus III-lymphadenopathy associated virus (HTLV-III/LAV), the virus which causes AIDS. The discovery of the virus and the widespread availability of serological tests greatly increase the ability to understand AIDS and to study the natural history of HTLV-III/LAV infection. PMID- 2990691 TI - Epidemiology of human lymphotrophic retroviruses: an overview. AB - The discovery of HTLV-I opened the way for the subsequent recognition of other human retroviruses. HTLV-I is linked to an aggressive T-cell cancer. The availability of a marker for the AIDS agent, HTLV-III, has substantially enhanced our ability to define the nature of the AIDS epidemic, its clinical and subclinical manifestations, and the spectrum of disease outcomes associated with this exposure. Of particular interest is the fact that there is substantial risk for AIDS and AIDS-related outcomes in virally infected individuals as detected by antibody positivity. The preliminary data from these well defined cohorts provide a basis for estimating the enormity of the AIDS epidemic as it is starting to emerge and provide an opportunity for the forward thinking health strategies necessary for dealing with a pandemic of such proportions. PMID- 2990693 TI - HTLV-III infection in homosexuals and hemophiliacs in Sweden. AB - Two hundred and three homosexual (HS) men and 114 hemophiliacs in Sweden were examined for serum antibodies to human T-lymphotropic virus type III (HTLV-III) and for alterations of T-lymphocyte subsets. Sera were screened for HTLV-III antibodies by an enzyme-linked immunosorbent assay and/or a dot immunobinding assay, and positive reactions were confirmed by Western blotting. HTLV-III antibodies were demonstrated in 13 of 13 (100%) HS men with acquired immune deficiency syndrome, in 63 of 67 (94%) HS men with persistent generalized lymphadenopathy, in 17 of 45 (38%) symptomatic HS men, and in 6 of 78 (8%) asymptomatic HS men but in none of 108 male blood donors. Seropositive HS men had significantly lower T4/T8 (helper/suppressor) cell ratios and T4 cell numbers than had seronegative HS men. Seronegative HS men had decreased T-cell ratios compared to controls but not decreased T4 cell numbers. Among hemophilia A patients, HTLV III antibodies were demonstrated in 40 of 48 (83%) cases treated with American factor VIII concentrate and in 17 of 29 (59%) cases treated with both American and Swedish concentrates but in none of 13 cases treated exclusively with Swedish factor VIII. Twenty-one hemophilia B patients treated with Swedish factor IX concentrates were all seronegative, whereas one of 3 hemophilia B cases treated with imported factor IX was seropositive. T4/T8 cell ratios were significantly lower in seropositive as compared to seronegative hemophilia A patients. PMID- 2990694 TI - Clinical and immunological findings in HTLV-III infection. AB - Clinical, immunological, microbiological, virological, and lifestyle parameters were followed in 200 homosexual men living in Finland. The subjects were seen at 3- to 6-month intervals starting in summer 1983. Human T-cell lymphotropic virus III (HTLV-III) antibodies detected by enzyme immunoassay and confirmed by Western blotting were seen in 18 (9%) of the cases. Initially two cases had acquired immunodeficiency syndrome, three had acquired immunodeficiency syndrome-related complex, three had lymphadenopathy syndrome, and ten were asymptomatic. During the follow-up two asymptomatic cases developed lymphadenopathy syndrome and three developed enlarged lymph nodes. Immunological studies revealed decreased T-helper cell values and/or T-helper/suppressor ratios in all clinical categories, the findings being more severe and progressive in the symptomatic cases. The finding most clearly distinguishing HTLV-III antibody positive cases from the antibody negative ones was a decreased response to a specific recall antigen, purified protein derivative of tuberculoprotein. Responses to mitogens phytohemagglutinin and pokeweed mitogen were also decreased, but to a lesser extent. The primary immunological defect associated with helper T-cell infection by HTLV-III thus seems to be loss of antigen specific immune responses. It is suggested that this is the result of previous antigenic stimulation of HTLV-III infected T-helper cells and that the cellular destruction is associated with initial mitotic activity. PMID- 2990695 TI - AIDS in Haitian-Americans: a reassessment. PMID- 2990696 TI - Detection of antibodies to HTLV-I and -III in sera from Japanese hemophiliacs. AB - Sera from 50 Japanese hemophiliacs were screened for antibodies to human T lymphotropic retrovirus types I and III (HTLV-I and -III). As a whole, antibody to HTLV-I, antibody to HTLV-III, and antibodies to HTLV-I and -III were detected in sera from 2, 17, and 6 hemophiliacs, respectively. Among them, two hemophiliacs developed acquired immunodeficiency syndrome who were positive for both antibodies to HTLV-I and -III in sera. All of the others were asymptomatic. Most of the blood products transfused into these hemophiliacs were imported from abroad, whence the source of HTLV-III infection presumably originated. However, since quite a high percentage of these antibody-positive hemophiliacs was positive for antibody to HTLV-I, even though they are native residents in HTLV-I nonendemic areas of Japan, some special factors may have participated in HTLV-I infection. These special factors should be investigated in the future. PMID- 2990697 TI - Unique pattern of HTLV-III (AIDS-related) antigen recognition by sera from African children in Uganda (1972). AB - Of 75 sera collected in the West Nile district of Uganda over a 1-year period between 1972 and 1973, 50 (66%) had antibody reactivity to human T-cell lymphotropic virus subgroup III (HTLV-III) at low titer levels. Sera were initially screened by HTLV-III enzyme linked immunosorbent assay and sera with values less than normal mean + 2 SD were removed from testing. The remaining sera were tested for positivity by an amplified Western blotting procedure which incorporated a three-layer immunoperoxidase procedure. Immunoglobulin reactive with HTLV-III Mr 24,000, 41,000, and 76,000 proteins were present in nearly all positive sera. The antibody status of this group was unlike any normal or acquired immunodeficiency syndrome-risk group previously tested. The high prevalence and relatively low titers suggest the detection as early as 1972 of a relative or predecessor of HTLV-III or of HTLV-III itself but existing in a population acclimated to its presence. It further suggests a likely African origin of HTLV-III. PMID- 2990698 TI - Comparative seroepidemiology of HTLV-I and HTLV-III in the French West Indies and some African countries. AB - The prevalence of antibodies detected by ELISA against human T-lymphotropic viruses, type I (HTLV-I) and type III (HTLV-III-LAV), is described in a comparative serosurvey in the French West Indies and African countries. The data confirm that the Caribbean basin is endemic for HTLV-I. In this region, HTLV-I antibody prevalence varied from 3.4% to 5.2% among blood donors and increased with age to reach a value of 33% among elderly people from the Dominica Island. In French Guyana, a South American country bordering the Caribbean sea, differences in antibody distribution across three ethnic groups (black Bonis, Indian Wayanas, and Hmongs from Asia) provide clues for investigation of the mode of HTLV-I transmission. Africa appears to be an endemic continent for HTLV-I and HTLV-III. For both viruses, the antibody prevalence exhibited an increasing gradient from northern to equatorial through Sudanic areas. These preliminary data by showing that Africa represents an endemic reservoir of HTLVs and, possibly, of other human retroviruses should stimulate further investigations on the natural history and the geographical origin of these viruses. PMID- 2990699 TI - Clinical spectrum of human retroviral-induced diseases. PMID- 2990701 TI - Clinical spectrum of HTLV-III in humans. AB - We have studied the clinical and laboratory manifestations of infection with human T-cell lymphotropic virus type III in various epidemiological cohorts. The spectrum of infection ranges from an asymptomatic but apparently contagious carrier state to severe immunodeficiency with opportunistic infections and neoplasms. Study of virus structure-function relationships and host response to viral infection in hosts with different clinical manifestations should provide strategies for therapeutics and vaccine development as well as enhance our understanding of the biology of human retroviruses. PMID- 2990700 TI - Spectrum of Kaposi's sarcoma in the epidemic of AIDS. AB - Kaposi's sarcoma (KS) is seen with increased frequency in the course of the epidemic of acquired immune deficiency syndrome. In this population, KS has manifested in an aggressive and more disseminated fashion as compared to the classical type. As the epidemic of acquired immune deficiency syndrome continues to spread and more cases of KS are evaluated, a distinct diversity in the clinical presentation and in the course of the disease as well as in variation in the prognosis and response to therapy is being observed. A preliminary description of the spectrum of KS in the epidemic of acquired immune deficiency syndrome is presented here. PMID- 2990702 TI - Clinical spectrum of infections in patients with HTLV-III-associated diseases. AB - Infectious complications are the cause of death in the overwhelming majority of patients with the acquired immunodeficiency syndrome (AIDS). These infectious complications are outstanding in severity and number. Infecting organisms are mainly those which take advantage of defective T-cell function. Many cause latent infections which can reactivate and cause disease in AIDS patients because of immunosuppression. Serological diagnosis of infections in AIDS patients is complicated because of the defect in B-cell function; rising antibody titers are not observed in most patients with infectious complications. The natural history of AIDS is one of repeated infections leading to a fatal outcome. However, treatment for most of the bacterial, fungal, and parasitic infections is effective. Treatment of some of these infections must be prolonged indefinitely because of the tendency to relapse. PMID- 2990703 TI - A classification of HTLV-III infection based on 75 cases seen in a suburban community. AB - Since 1981, 75 patients have been seen at our hospital with human T-cell lymphotropic virus type III (HTLV-III) infection. We have classified their clinical presentation into Groups 0 to 6. Groups 0 to 3 all have antibody to the Mr 41,000 protein of HTLV-III. Group 0 has no evident disease (9 patients), Group 1 has lymphadenopathy with or without exaggerated infection (16 patients), Group 2 has persistent lymphadenopathy with chronic hepatitis B surface antigenemia or profound hypergammaglobulinemia (7 patients), Group 3 has oral candidiasis with or without lymphadenopathy (7 patients). In Group 4 are acquired immunodeficiency syndrome (AIDS) adults or children (32 patients). Group 5 is a special classification for immunocompromised patients. Group 6 patients have lymphomas and Mr 41,000 protein antibody. Four children were classified separately. Three patients in Group 3 developed Group 4 disorders (AIDS). Four patients in Group 4 developed Group 6 disorders. HTLV-III infection spread in families (8 of 36), all from infected mothers to children. In 17 sexual partners, 6 were found to be infected. Five of 6 infected partners were homosexuals. We saw an inordinate number of transfusional AIDS (4 of 29) and 1 of 46 other disorders. Two infants also presented with severe intracranial defects, one with microcephaly and one with cranial calcifications and lucency. HTLV-III is spreading with alarming speed. PMID- 2990704 TI - Pediatric AIDS: a disease spectrum causally associated with HTLV-III infection. PMID- 2990705 TI - Lymph node pathology of HTLV and HTLV-associated neoplasms. AB - Human T-cell leukemia/lymphoma virus I can transform mature T-lymphocytes in vitro and is associated with the human T-cell cancer, adult T-cell leukemia/lymphoma. Adult T-cell leukemia/lymphoma is a distinct clinicopathological entity associated with leukemia, lymphadenopathy, hepatosplenomegaly, skin lesions, hypercalcemia, and lytic bone lesions. Although morphologically diverse it pursues an aggressive clinical course. Human T-cell leukemia/lymphoma virus III is associated with acquired immunodeficiency syndrome, which in its early stages shows follicular lymphoid hyperplasia; however, lymphoid atrophy is progressive and ultimately results in virtually total lymphoid depletion of lymph nodes. Patients with human T-cell leukemia/lymphoma virus III infections appear to have an increased risk of high grade B-cell lymphomas and perhaps Hodgkin's disease. PMID- 2990706 TI - Therapeutic approaches to patients with AIDS. AB - The immune systems of patients with acquired immunodeficiency syndrome are characterized by a profound defect in the number and function of helper/inducer T lymphocytes, particularly at the level of soluble antigen recognition. Due to this selective yet profound defect in the immune system, these patients are prone to recurrent severe opportunistic infections and Kaposi's sarcoma. While therapies exist for some of these complications of acquired immunodeficiency syndrome, no effective therapies exist for the underlying immune defect of this syndrome. Reviewed here are some of our recent attempts at immune reconstitution in acquired immunodeficiency syndrome, using either whole scale immune reconstitution through the use of lymphocyte transfers and bone marrow transplantation or enhancement of the remaining immune systems with the T-cell derived lymphokines interleukin-2 or immune gamma-interferon. In addition, recent advances in the therapy of Pneumocystis carinii pneumonia and disseminated cytomegalovirus disease are discussed. PMID- 2990707 TI - Unconventional vaccines: immunization with anti-idiotype antibody against viral diseases. PMID- 2990708 TI - Intensive induction chemotherapy for small cell anaplastic carcinoma of the lung. AB - The role of intensive induction chemotherapy in small cell cancer of the lung remains unclear. Twenty-eight newly diagnosed patients with small cell lung cancer were randomized to receive either high-dose or standard-dose cyclophosphamide, vincristine, and semustine in a trial where the dose of induction chemotherapy was the sole treatment variable. Complete responses (CR) were achieved in 57% of patients receiving high-dose therapy as compared to 21% of patients receiving standard-dose therapy (P less than 0.05). There was a higher rate of severe toxicity in the high-dose group (P less than 0.01). The overall median survival was not improved (36 weeks for the high-dose group vs 43 weeks for the standard-dose group); however, the median survival in patients achieving CR was prolonged (92 weeks for the high-dose group vs 50 weeks for the standard-dose group). These effects were most pronounced in patients with extensive disease; a CR after induction was achieved in five of nine patients treated with high doses but not in any of those treated with standard doses (P less than 0.05). A small group of patients appear to benefit by achieving a long term remission with intensive induction chemotherapy, but this effect may be offset by increased morbidity. PMID- 2990709 TI - Treatment of non-small cell bronchogenic carcinoma with vinblastine and mitomycin: a Southwest Oncology Group Study. AB - Forty-five patients with stage III M1 non-small cell bronchogenic carcinoma were treated with vinblastine (1 mg/m2 by iv bolus twice a day on 2 consecutive days) plus mitomycin (10 mg/m2 on Day 1). This treatment was repeated at 3-week intervals for three courses. Consolidation therapy with doxorubicin and cisplatin at doses of 50 mg/m2 each was administered to responders every 4 weeks for two courses, with subsequent vinblastine and mitomycin maintenance therapy every 6 weeks. Eleven partial remissions (24%) were achieved, with a median duration of remission of 16 weeks (range, 8-32) and a median survival of 19 weeks. No differences were seen in median duration of remission or survival by cell type or performance status. PMID- 2990710 TI - Combination chemotherapy followed by radiation therapy in patients with regional stage III unresectable non-small cell lung cancer. AB - A chemotherapy combination of cyclophosphamide, doxorubicin, and cisplatin (CAP) was administered to 30 patients with stage III M0 or M1 (supraclavicular nodes) unresectable non-small cell lung cancer before and after radiotherapy. All patients had mediastinal metastases and most had T2 or T3 primary lesions. The response rate (complete plus partial) after two cycles of CAP was 47%, which increased to 66% (24% complete response rate) following radiotherapy. The overall median survival from initiation of chemotherapy was 9 months. CNS relapse occurred in five (26%) of 19 responding patients who did not receive prophylactic cranial irradiation in the early part of the study. PMID- 2990711 TI - Cooperative trial of immunotherapy for nasopharyngeal carcinoma with transfer factor from donors with Epstein-Barr virus antibody activity. AB - A prospectively randomized, double-blind clinical trial was conducted to evaluate the effect of immunotherapy with transfer factor (TF) as an adjunct to radiotherapy of patients with stage III nasopharyngeal carcinoma (NPC). The TF was derived from normal young adults with a proven history of infectious mononucleosis and from normal blood donors with elevated antiviral capsid antigen antibody activity. TF prepared in this fashion was previously shown to convert the leukocytes of patients with NPC to a reactive state in vitro and when administered to NPC patients in vivo was associated with apparent slowing of tumor growth, marked lymphocytic infiltration of the tumor, and reconstitution of delayed cutaneous hypersensitivity reactions. From 1974 to 1977, 100 patients with NPC were entered in the study; one-half of the patients were treated with radiotherapy alone and one-half received radiotherapy and an 18-month course of TF immunotherapy. The patients were followed for at least 5 years. No significant difference in disease-free survival or survival was noted between the two groups of patients. The use of this particular preparation and dose schedule of TF in patients with NPC with regional disease was devoid of any anti-tumor activity. PMID- 2990712 TI - Increased cytosol glucocorticoid receptor content of leiomyosarcoma. AB - Glucocorticoid receptor content was determined in cytosol preparations of soft tissue tumors from 42 patients. Leiomyosarcoma had significantly elevated receptors in comparison to malignant melanoma, liposarcoma, malignant fibrous histiocytoma, and invasive desmoid. PMID- 2990713 TI - Effects of long-term verapamil treatment on blood pressure, cardiac hypertrophy and collagen metabolism in spontaneously hypertensive rats. AB - The effects of long-term treatment with verapamil on blood pressure, cardiac hypertrophy and collagen content, collagen concentration and prolyl hydroxylase activity were studied in spontaneously hypertensive rats (SHR). Verapamil administration (0.75 mg . ml-1 in drinking water) was commenced: to pregnant SHR 3 to 5 days before delivery and continued to the mothers and offspring during the nursing period; or to SHR at 10 weeks of age. Both groups were maintained on verapamil treatment up to the age of 45 weeks. Verapamil treatment significantly decreased blood pressure, heart rate and the ratio of ventricular weight to body weight in treated SHR. Verapamil did not significantly change the cardiac collagen concentration and prolyl hydroxylase activity. Since, however, the cardiac muscle mass was diminished by verapamil administration, treatment actually slightly reduced the collagen content of the heart. In the aorta collagen concentration was increased by verapamil treatment. Contrary to these results, minoxidil treatment was observed to increase the cardiac collagen concentration, content and prolyl hydroxylase activity in SHR. These results suggest that the factors governing myocardial connective tissue proliferation and regression may be independent of those governing muscle fibre hypertrophy and that particular drug actions on myocardial collagen metabolism must be taken into account. PMID- 2990714 TI - Effect of prazosin treatment on the cardiac sarcolemmal ATPase in failing heart due to mitral insufficiency in dogs. AB - Prazosin has been used in the treatment of congestive heart failure. It is, however, not known whether prazosin gives only haemodynamic benefit or if it also produces a decrease in the cardiac sarcolemmal Na+-K+-ATPase which has been reported to be increased in the failing heart. The present investigation deals with the effect of 3 months of prazosin treatment in dogs with 3 months of induced mitral insufficiency (MI) on the sarcolemmal Na+-K+-ATPase activity. The dogs were divided into four groups each comprising of five dogs. A--normal; B--3 months of MI; C--6 months of MI; D--3 months of prazosin treatment after 3 months of MI. Three months of MI produced a decrease in the dp/dt and an increase in the end-diastolic pressure of left ventricle but no change in the index of left ventricular contractility and cardiac index. Also there was no change in the sarcolemmal Na+-K+-ATPase. There was a significant decrease in the index of left ventricular contractility and cardiac index and an increase in the LVEDP associated with a significant increase in the left ventricular sarcolemmal Na+-K+ ATPase at 6 months of MI. Sarcolemmal Mg2+-ATPase of both ventricles increased after 6 months of MI the significance of which is not known as yet. There was no change in the sarcolemmal Na+-K+-ATPase of the nonfailing right ventricle. Prazosin treatment prevented the deterioration of the left ventricular contractility and function and also prevented the increase in the sarcolemmal Na+ K+-ATPase observed in failing heart.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990715 TI - Insulin-dependent cyclic AMP turnover in isolated rat adipocytes. PMID- 2990716 TI - Role of hydroxyl radical in neutrophil-mediated cytotoxicity. PMID- 2990717 TI - [Recent advance in research on the incidence of paralysis following immunization with live oral polio vaccine]. PMID- 2990718 TI - [An epidemic of hand-foot-and-mouth disease due to Coxsackie A16 in Tianjin City]. PMID- 2990719 TI - [Sero-epidemiologic study of rotavirus infection]. PMID- 2990720 TI - [Early diagnosis of viral hepatitis A--study on the use of ELISA in the detection of specific IgM antibody]. PMID- 2990721 TI - Endorphin patterns within the headache spectrum disorders. AB - The role of opioid peptides in modulating the nervous system adaptability has been demonstrated recently; proopiomelanocortin (POMC)-related peptides, in particular, serve in pain perception, in adaptation to stress, and in modulating higher brain functions. Primary headaches, besides pain, involve neuroendocrine/autonomic/adaptive processes as well as mood and personality factors. The view that primary headaches can be taken as a possible model of POMC related peptides dysfunction led us to evaluate the resting plasma and CSF peptide levels and their plasma changes in response to various stimuli affecting their release. The data obtained from basal and dynamic studies agree with the concept that primary headaches are sustained by opioid system disturbance. In particular the reduced release of endogenous opioids by anterior pituitary in response to physical, endocrine or pharmacological stimuli agrees with a weak adaptive ability of headache sufferers. This impairment of endorphin responsiveness could play a key role in headache susceptibility to environmental stimuli. Primary headaches constitute a wide, intriguing field, including several subgroups bordering on "ischemic" and behavioral/affective disorders. The development of neuroendocrine techniques could be a useful means for supporting the clinical criteria identifying subpopulations of headache sufferers. PMID- 2990722 TI - Estrogen changes as a critical factor in modulation of central opioid tonus: possible correlations with post-menopausal migraine. AB - The effects exerted by ovarian steroids on the modulation of opioid activity were investigated in post-menopausal migraine sufferers and in healthy controls. In order to evaluate central opioid tonus, plasma LH rise after naloxone injection was measured, bearing in mind the tonic inhibition of endogenous opioid on hypothalamic LH-RH. There was no response of plasma LH to naloxone in post menopausal women or in patients submitted to ovariectomy in fertile life. When the subjects underwent a sequential estrogens + progestagens therapy, such a response was noted from the first month of treatment; progestagens alone were ineffective. The same phenomena were also evident in post-menopausal migraine sufferers. These data indicate that ovarian steroids modulate the activity of opiate receptors in both healthy women and migraine sufferers. Interestingly, replacement therapies through ovarian steroids restored the activity of central opioid tonus in patients affected by migraine. PMID- 2990723 TI - Activation of the transformation potential of the cellular fps gene. AB - Chicken cellular-fps (c-fps) sequences were substituted for viral-fps (v-fps) sequences in two retroviral genome structures, one that expressed a c-fps gene product that was indistinguishable from the normal c-fps gene product expressed in chicken bone marrow cells, and another that expressed a gag-fps fusion protein. When c-fps gene sequences (without linked gag gene sequences) were expressed at high levels in a viral vector, no transformation of fibroblasts was detected. It was previously demonstrated that the corresponding v-fps sequences could transform fibroblasts. When the same c-fps sequences were expressed in a form linked to gag gene sequences, transformation of fibroblasts and induction of tumors were observed. The data suggest that the c-fps gene product lacks transformation potential by itself even when overexpressed and that the transformation potential of the c-fps gene can be activated by either mutation (or mutations) in the fps coding region or by fusion with viral gag gene sequences. PMID- 2990724 TI - Transactivation of a bovine papilloma virus transcriptional regulatory element by the E2 gene product. AB - We have mapped a transcriptional regulatory sequence within the 1.0 kb noncoding region of the bovine papilloma virus (BPV-1) genome, using an enhancer dependent expression vector for chloramphenicol acetyltransferase. This transcriptional regulatory element works independently of position and orientation, and its function is significantly augmented in BPV-1 transformed C127 cells and in monkey CV-1 cells acutely transfected with plasmids expressing BPV-1 early gene products. Using defined deletion mutants of the BPV-1 DNA and full-length viral cDNAs expressed from an SV40 early promoter, we demonstrate that the expression of this trans-activating factor maps to the 3' open reading frames of the viral transforming region. A premature termination codon engineered into the E2 ORF eliminates expression of this diffusible transactivation function establishing the E2 gene product as the diffusible trans-activating factor. PMID- 2990726 TI - Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site. AB - The positive regulator encoded by phage lambda gene Q is a transcription antiterminator that affects RNA polymerase initiating at the phage late gene promoter, but not at other promoters. We show that this nucleotide-sequence specific interaction of Q protein and RNA polymerase can occur while the enzyme is pausing after 16 nucleotides of the late gene transcript have been made. Furthermore, Q protein chases RNA polymerase from this early pause site, so that it both recognizes the enzyme and changes its transcription properties at this site. We suggest that the ability of Q-modified RNA polymerase to escape this pause reflects the change that allows it to go through terminators. We also show that NusA protein is required for efficient Q protein activity in vitro. PMID- 2990725 TI - Viral oncogenes. PMID- 2990727 TI - A structural basis for S1 nuclease sensitivity of double-stranded DNA. AB - A protonated form of a cloned simple repeating DNA sequence d(TC)n X d(GA) is detectable in equilibrium with the usual Watson-Crick base-paired form at pHs up to 7. This form is anomalously sensitive to a variety of single-strand-specific endonucleases. The observed pH dependent protection of N-7 of dG residues within the insert suggests that these residues are either Hoogsteen or reverse Hoogsteen base-paired to protonated dC residues of the polypyrimidine strand. A structure in which dA:dT Watson-Crick base pairs alternate with Hoogsteen syndG:dCH+ pairs appears to be the most stereochemically acceptable structure consistent with the chemical properties of this protonated DNA. Protonated d(TC)n X d(GA)n interacts with an anti-Z DNA antibody raised against brominated d(GC)n X d(GC)n. PMID- 2990728 TI - Molecular cloning of engrailed: a gene involved in the development of pattern in Drosophila melanogaster. AB - The engrailed gene acts early in Drosophila embryogenesis and plays an essential role in the processes that establish and maintain the repeating segmental pattern. To begin molecular analysis of the role of the engrailed gene in embryonic pattern formation, we used a chromosomal walk to clone genomic sequences that encompass the locus, and have physically mapped the positions of 15 engrailed mutations. The positions of engrailed rearrangement mutations indicate that the engrailed complementation unit includes a minimum of 70 kb. The locus can be divided into two regions. Rearrangement mutations interrupting the centromere proximal 50 kb of the locus result in embryonic lethality while mutants altered in the distal 20 kb of the locus survive to show morphological abnormalities in several adult segments. It appears that long-range cis interactions play a role in the function of the engrailed gene. PMID- 2990729 TI - Retroviral DNA integration. PMID- 2990730 TI - The isolation and nucleotide sequence of a cDNA encoding the T cell surface protein T4: a new member of the immunoglobulin gene family. AB - The surface glycoproteins T4 and T8 define different functional subsets of T lymphocytes and may act as recognition molecules mediating appropriate interactions between the T cell and its target. Previously we employed gene transfer and subtractive hybridization to isolate a T8 cDNA; now we have isolated and sequenced a cDNA clone encoding the T4 molecule. The deduced protein sequence reveals that T4 is an integral membrane protein that shares significant amino acid and structural homologies with members of the immunoglobulin supergene family. The overall structure of T4 consists of an N-terminal variable (V)-like domain, a joining (J)-like region, a third extracellular domain, a membrane spanning region homologous to class II MHC beta-chains, and a highly charged cytoplasmic domain. Comparison of the protein sequences deduced from the T4 and T8 cDNAs reveals structural similarities consistent with their postulated role as recognition molecules, as well as differences suggesting that the two proteins recognize different structures on the target cell. PMID- 2990731 TI - Regulation of amphibian oocyte maturation. AB - Xenopus oocyte maturation is a model system for studying the control of cell proliferation and the regulation of the cell cycle. Addition of progesterone or insulin to oocytes releases a G2 block and stimulates progression through meiosis to an unfertilized egg. The release of the G2 block is a consequence of a decrease in cAMP mediated entirely or in part by an inhibition of adenylate cyclase. The mechanism of cyclase inhibition involves a membrane steroid receptor controlling the rate of guanine nucleotide exchange. Subsequent events include an increase in intracellular pH and the phosphorylation of ribosomal protein S6. The latter event may play a role in translational control of maturation. Late events in maturation involve the appearance of the maturation-promoting factor (MPF), a cytoplasmic protein responsible for causing nuclear envelope breakdown, chromosome condensation, and spindle formation. MPF oscillates in meiotic and mitotic cell cycles. The events caused by MPF can now be obtained in crude extracts with retention of cell cycle control by calcium, providing a framework for rapid progress in characterizing MPF and its regulation. PMID- 2990732 TI - Somite chondrogenesis: alterations in cyclic AMP levels and proteoglycan synthesis. AB - Cyclic AMP (cAMP) levels have been shown to have a positive influence on chondrogenesis in limb buds and pelvic cartilage. In the present study the level of cAMP was measured during somite chondrogenesis in vitro and found to decrease from 1.38 pmol/micrograms DNA on day 0 to 0.9 pmol/micrograms DNA on day 6. Inclusion of notochord with somites caused a marked reduction, with levels decreasing from 1.41 pmol/micrograms DNA on day 0 to 0.36 pmol/micrograms DNA on day 6. Concurrently, the incorporation of radioactive sulfate into sulfated glycosaminoglycans increased from day 3 to day 6 by 38% in somite and 77% in somite-notochord explants. The aggregation of proteoglycans was analyzed by gel chromatography and found to increase with a corresponding decrease in cAMP levels. The results indicate that a decrease in cAMP levels may be necessary for chondrogenic expression in somites. PMID- 2990733 TI - Regulation of B-lymphocyte production in the bone marrow: role of macrophages and the spleen in mediating responses to exogenous agents. AB - B-Lymphocyte production in mouse bone marrow can be stimulated by administering a variety of foreign materials in vivo. The nature and location of cells mediating this effect have now been studied, using assays of lymphocyte renewal and pre-B cell proliferation. Pretreatment of mice with silica, to depress macrophage function, abolished the stimulation of small lymphocyte renewal produced by administering either sheep red blood cells (SRBC) or mineral oil and reduced the response to bovine serum albumin. The response was still abolished when silica was given 6 or 24 hr, but not 48 hr, after SRBC. Splenectomy prevented the stimulation of marrow lymphocyte renewal when performed either 4 weeks before or up to 72 hr after SRBC injection. The stimulation of pre-B-cell proliferation was similarly prevented by pretreatment with either silica or splenectomy. The results indicate that the wave of increased B-lymphocyte production after SRBC injection depends for the first 2-3 days upon silica-sensitive, spleen-dependent mechanisms, suggesting an early mediation by splenic macrophages. Primary B lymphocyte production in vivo may thus normally be stimulated by exposure to external environmental agents acting indirectly on bone marrow B-cell progenitors via cellular reactions in peripheral lymphoid tissues. PMID- 2990734 TI - Lymphocyte activation by purified HLA-DR molecules fused into autochthonous "stimulating cells". AB - Affinity-purified Ia molecules derived from the Daudi cell line were reconstituted into vesicles with Sendai virus envelope glycoproteins. These vesicles inserted into human peripheral leukocytes could induce stimulation of autologous lymphocytes, as measured by thymidine uptake, 6 days later. It is suggested that this method could provide a means to study allostimulation at the molecular level. PMID- 2990735 TI - Human epidermal cells are more potent than peripheral blood mononuclear cells for the detection of weak allogeneic or virus-specific primary responses in vitro. AB - Human epidermal cells (EC) and peripheral blood mononuclear cells (PBMC) have been used as antigen-presenting cells in allogeneic reactions or in self restricted antiviral responses. Comparison of results from both cell types indicates that: (1) EC were better stimulators of primary proliferative responses in all the antigenic systems tested. (2) In secondary reactions, EC and PBMC functioned similarly for allogeneic responses, while a weak but significant difference could be observed in both (HSV1 or influenza A) virus-specific reactions. (3) By comparing pairs of HLA-identical mixed lymphocyte reaction (MLR)-negative siblings, positive responses were observed in several different families when lymphocytes of potential bone marrow donors were stimulated by EC of the recipient. This suggests that EC might be useful in detecting relatively weak proliferative responses in a number of antigenic systems, but especially in primary reactions against viral or putative minor histocompatibility antigens. (4) Despite this stronger antigen-presenting capacity in proliferative responses, EC induced lower levels of cytotoxic T lymphocyte (CTL) reactions than PBMC, not only in allogeneic responses but also in virus-specific self-restricted reactions. PMID- 2990736 TI - The role of HLA antigens in the control of the cytotoxic T-cell response to Epstein-Barr virus: a family study. AB - Epstein-Barr (EB) virus-specific, HLA-restricted cytotoxic T-cell populations have been generated in vitro from each member of a family by cocultivating peripheral blood mononuclear cells with autologous EB virus-transformed B cells, the resulting effector cells being expanded as interleukin 2-dependent T-cell lines. The cytotoxicity of each of these effector populations was tested on a large panel of EB virus-transformed target cells of known HLA type, so that the particular HLA antigens which acted as restricting elements for each cytotoxic population could be identified. There was a consistent pattern within the family of preference for certain HLA class I antigens as restricting elements of the virus-specific T-cell response. Extensive functional analysis showed that, in addition to the virus-specific lysis, each effector population mediated a cross reactive lysis against target cells prepared from certain HLA-mismatched individuals. This cross-reactivity appeared to be directed against HLA class I alloantigens and occurred irrespective of the EB virus genome status of the target cells. Effector T-cell lines derived from different family members but with virus-specific lysis predominantly restricted through the same HLA antigen showed similar patterns of concomitant allo-cross-reactivity. This suggests that antigenic mimicry of virally altered self by alloantigens is a genuine phenomenon which may be important in channelling the human cytotoxic T-cell response to a virus through preferred self-HLA determinants. PMID- 2990737 TI - Activation of murine thymocyte subpopulations by the oxidizing mitogens: evidence for a sialylated, mitogen-reactive site on both PNA-positive and PNA-negative cells. PMID- 2990738 TI - Segregation of the NK-sensitive phenotype in human x mouse somatic cell hybrids reveals separate genetic control of recognition and postrecognition determinants. AB - In an attempt to investigate the nature of tumor cell-derived membrane surface determinants involved in natural killer cell (NK) recognition or postrecognition events, we have constructed human X mouse interspecies somatic cell hybrids. Highly NK-sensitive (NKs) human tumor cells were fused with NK resistant (NKr) mouse fibroblasts (LMTK-) in polyethylene glycol and selected in hypoxanthine/aminopterin/thymidine medium and ouabain. Hybrids generated from NKs erythroleukemia cells (K-562) or NKs retinoblastoma cells (Y-79) with LMTK- displayed an intermediate NK-sensitive phenotype. One Y-79 X LMTK- hybrid (YL-22) retained a high level of susceptibility to NK binding and cytolysis, as determined by 51Cr release and in cold-target inhibition assays. On the other hand, human NKr RAJI cells generated NK-resistant hybrids when fused with LMTK- fibroblasts. Four hybrids (KL-12, YL-2, YL-22, and YL-43) displaying consistent NK sensitivity were subsequently cloned by limiting dilution. Various hybrid clones derived from the KL-12 hybrid (K-562 X LMTK-) demonstrated a range of NK sensitive phenotypes. However, the uncloned KL-12 and most cloned lines derived from this hybrid competed against 51Cr-labeled K-562 targets as well as unlabeled K-562 parental cells, regardless of their NK-sensitive phenotype. These findings raise the possibility that chromosomal segregation may be affecting a postbinding step in this hybrid system. The NK-sensitive hybrids exhibited a limited number of human chromosomes as assessed by quinacrine banding. Furthermore, human transferrin receptor (TfR) expression, as monitored by flow cytometry using the B3/25 monoclonal antibody, demonstrated no clear correlation with NK sensitivity or competitive ability in either KL or YL hybrid clones, thus arguing against the involvement of the TfR in human NK recognition. These results suggest that the NK sensitive phenotype in human tumor cells may be regulated by genes encoded by a limited number of human chromosomes. PMID- 2990739 TI - How Golgi-associated glycosylation works. PMID- 2990740 TI - Oxygen consumption by tumor cells during membrane vesicle shedding. AB - The incubation of mastocytoma P815 cells at low temperature (0 degrees C/1-2 hr), with a subsequent shift to greater than or equal to 20 degrees C results in the formation and shedding of membrane vesicles from the tumor cell surfaces. This process, when occurring at physiologic temperature (37 degrees C), mimics the morphological and membrane permeability changes occurring during T-lymphocyte mediated cytolysis of tumor cells. The latter is an oxygen dependent event, but it is not known whether this requirement is at the effector T cell or at the tumor cell level. The present study investigated the oxygen consumption rates of mastocytoma P815 cells induced to shed membrane vesicles by a temperature shift (0 degrees C/1-2 hrs----greater than or equal to 20 degrees C). Results showed that cells undergoing the membrane vesicle shedding process had significantly higher oxygen requirements than control non-shedding cells. Inhibition of the shedding process with deuterium oxide and hexylene glycol, reduced the oxygen consumption rates of low temperature treated cells to the level of control cells. The oxygen consumption rates of the latter were unaffected by these microtubule stabilizing agents. These data indicate that the oxygen required for immune T cell mediated lysis of tumor cells may be at the target tumor cell level. PMID- 2990742 TI - [Chemodectoma causing narrowing of the pharyngeal lumen]. PMID- 2990741 TI - [Preparation of 1-substituted 4-dialkylaminoalkylamino-3-methyl-1H -pyrazole(3,4 b)quinolines with an antiviral effect]. PMID- 2990744 TI - [The mechanism of receptor transmission. III]. PMID- 2990743 TI - [An intracranial tumor in an unusual location associated with Cushing's syndrome]. AB - In a 58 year-old woman with peracute Cushing's syndrome high levels of ACTH were proved by RIA method. Autopsy showed normal pituitary gland in addition to a voluminous tumour situated in planum sphenoidale and lamina cribrosa ossis ethmoidalis. Tumour cells with epithelial features produced ACTH which was found by an indirect immunoperoxidase technique. Tumour was classified as ectopic corticotropic pituitary adenoma. PMID- 2990746 TI - Stereoselective reactions. VIII. Stereochemical requirement for the benzylic oxidation of lignan lactone. A highly selective synthesis of the antitumor lignan lactone steganacin by the oxidation of stegane. PMID- 2990745 TI - [Clinical significance of detecting prenatal infections in malformed fetuses]. PMID- 2990748 TI - Prevention and control of herpesvirus diseases. Part 1. Clinical and laboratory diagnosis and chemotherapy. A WHO meeting. AB - The herpesvirus diseases are increasing in importance as a public health problem throughout the world. Members of the human herpesvirus family are global in distribution and infect 60-95% of the world's population, both in developed and in developing countries. Illnesses associated with herpesviral infections vary from simple blisters to deadly encephalitis. In numerical terms, primary cytomegalovirus infection is a more common cause of congenitally acquired disease than primary rubella and results in severe handicap. The apparent increasing incidence of genital herpes, which may induce perinatal infections associated with a high rise in perinatal morbidity and mortality, is a cause of concern. Neonatally acquired infection has a high mortality and many of the survivors have permanent sequelae. Herpes simplex virus, cytomegalovirus and varicella-zoster virus reactivations frequently lead to death in patients undergoing immunosuppressive therapy. Close association between herpes simplex infection and cervical neoplasia has been reported.Unlike other viruses, the herpesviruses usually remain latent throughout life. Viral reactivations, induced by various factors (hormonal, environmental, physical or chemical agents), may result in a large variety of illnesses, with high prevalences in both developing and developed countries.Advances are currently being made in diagnostic, preventive and therapeutic approaches to the human herpesviruses. There are new methods for rapid disease diagnosis and for epidemiological investigations, including the rapid analysis of viruses and antibodies. New antiherpetic drugs have been licensed in some countries, and vaccines are undergoing trials.This report summarizes the information currently available and makes recommendations to enhance progress in the control of diseases caused by the herpes-viruses. PMID- 2990747 TI - The clinical significance of minimal breast cancer: a pathologist's viewpoint. AB - We can draw the following conclusions about minimal breast cancer: The concept of minimal breast cancer as a stage of cancer that is 95% curable is a valid one, if minimal breast cancer is defined by strict parameters. Both 0.5 and 1.0 cm have been defined as the upper limit of size for minimal invasive cancer. Some data indicate that 0.5 cm is the preferable dividing line and that 1-cm cancers are no longer minimal. Other data suggest that the most important factor is axillary lymph node status. One-centimeter cancers are probably 95% curable if axillary lymph nodes are negative. Cancers of 0.5 cm and smaller in size are probably not 95% curable if axillary lymph nodes are involved. Carcinoma in situ appears to be highly curable, even if axillary lymph nodes are involved. Minimal breast cancer should include lobular carcinoma in situ (lobular neoplasia) and ductal carcinoma in situ regardless of nodal status, and (tentatively) invasive carcinoma smaller than 1 cm in total diameter, if axillary lymph nodes are not involved. Many cases of minimal breast cancer are asymptomatic. If special screening is not used, less than 10% of women with breast cancer will be at the minimal stage when diagnosed. Screening programs can increase this ratio to as much as one third of patients, perhaps even more. While serious questions about cost effectiveness of mass screening remain, screening programs appear to represent the best way of detecting minimal breast cancer. Screening programs should include careful history and physical examination, of course. The role of mammography is still controversial. It is probable that at least 50% of all minimal cancers would be missed without mammography. After a period of significant worry about the risk of radiation, opinion seems to be changing now and many authors are willing to accept the fact that mammography is of more benefit than risk for younger women. The HIP study would indicate that the risk/benefit ratio becomes favorable at age 50. Many authorities would now comfortably include mammography in the screening of women age 40 or older. Some authors believe that the benefits of mammography outweight the risks for patients of all ages. This question needs to be tested, and several randomized prospective clinical trials now in progress are doing just that. The legitimate worry over the risks of mammography should not obscure a very important fact.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990749 TI - Drug monitoring of etoposide (VP16-213). I. A combined method of liquid chromatography and mass spectrometry. AB - Drug monitoring is performed by means of sample extraction, sample purification by high-performance liquid chromatography (HPLC), and sample detection by time-of flight mass spectrometry. This mass spectrometry utilizing 252Cf fission fragment induced ionization and desorption of nonvolatile compounds is suitable as a universal, nondestructive detector in HPLC. Liquid chromatography and mass spectrometry are combined, so that mass analysis can be operated online and offline to the fractional sampling of the effluent and the samples can still be recovered. As an alternative to HPLC separation, samples can be purified by thin layer chromatography (TLC), resulting an offline TLC + MS combination. Preliminary pharmacokinetic data for etoposide (VP16-213) together with calibration data are presented, and are discussed with reference to the sensitivity and detection limit of the new experimental method. PMID- 2990750 TI - Doxifluridine (5'-dFUrd) in patients with advanced colorectal carcinoma. A phase II study. AB - Infusion of 5'-dFUrd (2.0-3.0 g/m2 over 1 h on days 1-5 every 3rd week) resulted in one partial response in 21 patients with advanced and progressing colorectal cancer. No patient had received chemotherapy before the 5'-dFUrd trial. Hematological and gastrointestinal toxicity were generally mild. In 4 patients peripheral neurotoxicity was diagnosed during treatment, whereas transient cerebellopathy was observed in one. Cardiac side effects (repeated angina pectoris following 5'-dFUrd infusion) led to discontinuation of treatment after two courses in one patient. It is concluded that 5'-dFUrd at the above doses is not superior to conventional 5-FU treatment in colorectal cancer. Neurological and cardiac side effects are rare but may be a problem in individual patients. PMID- 2990751 TI - Phase II study of a combination of mitomycin, doxorubicin and cisplatin in advanced sarcomas. AB - In all 63 patients were treated monthly with a combination of mitomycin, doxorubicin, and cisplatin, and 27 (43%) experienced objective regression of their advanced sarcomas during a 3-month trial. The observed regression rate is numerically higher than any previously observed at our institution. PMID- 2990753 TI - Poly (ADP-ribose) metabolism in alkylated mouse L5178Y cells. AB - Poly ADP-ribosylation of two mouse lymphoma cell lines, L5178Y (LS) and the radiation and alkylating agent resistant derivative AII, was investigated by uptake of [3H]NAD by permeabilised cells into acid-precipitable material that was sensitive to phosphodiesterase but insensitive to DNase and RNase. Basal activities in both lymphoma lines were 3-4-fold greater than in mouse L1210 leukaemia cells. However, total endogenous poly (ADP-R) polymerase activity in both L5178Y cell lines, stimulated by a large excess of DNase in the presence of Triton X-100, was less than half the activity in L1210 cells. Doses of N-methyl-N nitrosourea (MNU) that produced 20-50% survival of colony-forming units increased poly (ADP-R) in both lymphoma lines by only 25% compared with 377% in L1210 cells when synthesis was measured immediately after a 30-min exposure of MNU. During the first 24 h after MNU AII cells produced a peak of activity that was not seen with LS cells. A second peak was seen in both cell lines between 24 and 48 h following MNU. Concentrations of 3-aminobenzamide (3AB) above 2.5 mM inhibited colony-forming ability of lymphoma cells and equally inhibited uptake of [14C]formate into protein, RNA and DNA indicating that 3AB behaves as a general metabolic poison. Concentrations of 3AB in the toxic range of 3-10 mM inhibited poly (ADP-R) synthesis but no degradation of the polymer was observed. Non-toxic concentrations of 3AB potentiated cell killing by MNU to a similar degree in both lymphoma cell lines. In conclusion, we have found little evidence to support the hypothesis that the differential sensitivity of LS and AII is related to poly ADP ribosylation. Compared with other mouse cells, L5178Y cells appear deficient in poly (ADP-R) polymerase and poly (ADP-R) glycohydrolase activities. PMID- 2990752 TI - The effect of cisplatin on renal ATPase activity in vivo and in vitro. AB - The effect of cisplatin on ATPase activity was determined in vitro and in vivo to investigate the correlation between nephrotoxicity and the inhibition of ATPase activity by cisplatin. Purified Na,K-ATPase was preincubated for 0-240 min with cisplatin at concentrations of 50-800 microM in vitro before the determination of enzyme activity. Although ATPase activity was reduced by cisplatin, either a high concentration of cisplatin (280 microM) or a long period of preincubation (160 min) with cisplatin was required to obtain 50% inhibition of ATPase activity. Similar in vitro experiments using kidney homogenate from female Sprague-Dawley rats instead of purified Na,K-ATPase were performed. Activity of Na,K-ATPase in rat kidney homogenate was inhibited by 50% after 110 min preincubation with 800 microM cisplatin or 160 min preincubation with 400 microM cisplatin. Female Sprague-Dawley rats were given 5, 7 or 10 mg/kg of cisplatin IV and BUN level, ATPase activity and Pt concentration in kidney homogenate were evaluated 1 h, 6 h, 1 day, 3 days, and 5 days after cisplatin injection. In rats given 10 mg/kg cisplatin a significant increase of BUN was observed on days 1, 3, and 5. In rats treated with 5 or 7 mg/kg of cisplatin BUN was increased on days 3 and 5. Normal ATPase activity, however, was preserved until day 3 at all doses. The highest concentration of Pt observed in kidney tissue was 19.3 micrograms/g tissue. This value was insufficient to inhibit ATPase activity significantly in vitro. Thus, it seems unlikely that the inhibition of ATPase activity is the cause of nephrotoxicity, although cisplatin can affect ATPase activity. PMID- 2990754 TI - Adenosine diphosphate ribosyl transferase in marrow cells of patients with acute myeloid leukemia is related to differentiation and drug sensitivity. AB - The levels of adenosine diphosphate ribosyl transferase (ADPRT) have been quantified in Ficoll-Isopaque isolated marrow cells from 36 patients with acute myeloid leukemia (AML). The in vitro growth pattern in agar at diagnosis was also determined, and in 16 patients the in vitro drug sensitivity of the clonogenic cells (CFU-GM) to cytosine arabinoside and daunorubicin was measured. The ADPRT activities of the various marrow cell preparations correlated to the morphological diagnoses, in vitro growth patterns, in vitro drug sensitivities to cytosine arabinoside, and to the prognoses of the AML patients. Hence, ADPRT may be a useful marker for the pathophysiology associated to AML. PMID- 2990755 TI - Role of hormones on the induction of retinoic acid binding protein in mouse mammary gland organ culture. AB - Influence of various hormones on the induction of cellular retinoic acid binding protein (CRABP) was investigated in the mouse mammary gland organ cultures. Thoracic pairs of mammary glands from the BALB/c mice were cultured for seven days in the presence of various hormones in CMRL medium at 37 degrees C under 50% O2, 5% CO2 and 45% N2 atmosphere. There was a modest increase of mammary CRABP by insulin (I) + prolactin (P), however, addition of progesterone (Pg) or estrogen (E) + Pg to the medium resulted in a dramatic increase in the CRABP. Aldosterone (A) + hydrocortisone (F), in addition to I + P, which promotes differentiation to an extent similar to that of I + P + E + Pg did not have any influence on the induction of CRABP. These results indicate that prolactin and/or Pg in the medium can increase the concentration of CRABP in the mammary gland in vitro. From the results presented in this report, as well as previous work by other investigators, it is concluded that the biological response to retinoids in the mammary tissues cannot be correlated with the absolute concentration of CRABP in the cells. However, the biological response may be dependent upon both the ability of the target organ cells to metabolize the retinoid and to have minimal concentration of CRABP for binding to the active metabolite. The functional significance of hormone-induced CRABP is presently unknown. PMID- 2990756 TI - Hepatic neoplastic nodules, adenofibrosis, and cholangiocarcinomas in male Fisher 344 rats fed corn naturally contaminated with Fusarium moniliforme. AB - Twelve male Fisher 344 rats were fed corn naturally contaminated with Fusarium moniliforme. This corn sample was obtained from feed being fed horses during an outbreak of leukoencephalomalacia. All rats necropsied from 123 to 176 days post feeding had multiple hepatic nodules and pale depressed hepatic areas. Histological examination revealed multiple hepatic neoplastic nodules and large areas of adenofibrosis and cholangiocarcinomas. The corn only diet was deficient in many nutrients including choline and methionine. Aflatoxins at a level of greater than 0.9 p.p.b. were not detected. A discussion is presented regarding the significance of these lesions and possible relationship to the contaminating fungus and the choline, methionine deficient diet. PMID- 2990757 TI - Identification of cembratriene-4,6-diol as antitumor-promoting agent from cigarette smoke condensate. AB - Cigarette smoke condensate (CSC) was separated into several fractions and each was tested for an inhibitory effect on the early antigen (EA) of Epstein-Barr virus (EBV) which can be induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells. Two diastereoisomers of 2,7,11-cembratriene-4,6-diol (alpha- and beta CBT) were isolated from the neutral fractions of CSC and these showed potent inhibitory effects on the induction of EBV-EA by TPA. The doses of alpha- and beta-CBT required for 50% inhibition of EBV-EA induction by TPA were 7.7 and 6.7 micrograms/ml, respectively, which are comparable with those of retinoic acid, a potent inhibitor of induction of epidermal ornithine decarboxylase (ODC) activity and tumor promotion by TPA in mice. Application of alpha- and beta-CBT to mouse skin prior to treatment with TPA inhibited TPA-induced ODC activity. The degree of inhibition was dependent on the dose and application of 16.5 mumol/mouse of alpha- and beta-CBT resulted in a 50 and 40% reduction, respectively, of the maximum of the ODC activity induced as a result of treatment with TPA. In initiation-promotion experiments, alpha-CBT markedly inhibited the promoting effect of TPA on skin tumor formation in mice which were initiated with 7,12 dimethylbenz[a]anthracene, but beta-CBT was found to be less effective. Application of 3.3 mumol of alpha-CBT 40 min prior to treatment with TPA (1 microgram) resulted in a 53% reduction in the number of papillomas per mouse. Our present data suggest that EBV-EA inhibition assay using Raji cells is effective for the first screening of inhibitors of tumor promotion, and provide evidence that CSC contains antitumor-promoting agents in addition to carcinogenic and tumor-promoting agents already reported. PMID- 2990758 TI - Localization of beta 1- and beta 2-adrenergic receptors in rat kidney by autoradiography. AB - beta-Adrenergic receptor subtypes were localized and differentiated in rat kidney slices by in vitro autoradiography using the nonselective beta-antagonist [125I]iodocyanopindolol in the presence of the selective agents betaxolol (beta 1) and zinterol (beta 2). [125I]Iodocyanopindolol binding to kidney sections in the presence of these agents could be differentiated into high- and low-affinity components as predicted by the subtype selectivity of the compounds. Autoradiography revealed that: beta-adrenergic receptors were highly concentrated within the renal cortex, especially in glomeruli and juxtaglomerular granule cells, and to tubular sites in the cortex and medulla; [125I]iodocyanopindolol labeling of the juxtaglomeruluar granule cells was abolished at lower concentrations (10(-8) M) of betaxolol than was the labeling of glomeruli (10(-6) -10(-4) M); and zinterol had little effect on labeling of juxtaglomerular granule cells and glomeruli unless high concentrations were used, whereas the tubular labeling in medulla was much more sensitive to incubation with zinterol. These results indicate that beta 1- and beta 2-adrenergic receptor subtypes are differentially distributed within the kidney: beta 1, predominantly contained in juxtaglomerular granule cells and glomeruli, and beta 2, predominantly in medullary tubules. In vitro autoradiography provides a useful means to examine different receptor populations in discrete tissue areas. PMID- 2990759 TI - Renal nerve stimulation causes alpha 1-adrenoceptor-mediated sodium retention but not alpha 2-adrenoceptor antagonism of vasopressin. AB - Renal alpha 2-adrenoceptor stimulation by epinephrine infusion reverses cyclic adenosine monophosphate-mediated effects of vasopressin on sodium and water excretion. We used this response to determine whether renal nerve stimulation can activate renal alpha 2-adrenoceptors in the non-recirculating isolated perfused rat kidney (Krebs-Henseleit solution; 3.5 g/100 ml Ficoll; 1 g/100 ml albumin; 36 degrees C; propranolol 100 nM). In the presence of alpha 1-adrenoceptor blockade with prazosin (30 nM) alpha 2-adrenoceptor stimulation with epinephrine reversed the cyclic adenosine monophosphate-mediated effects of vasopressin on sodium (P less than 0.05) and water (P less than 0.05) excretion. Subthreshold (for vasoconstriction) renal nerve stimulation (10 V; 1 msec; 0.65 +/- 0.10 Hz) failed to alter the effect of vasopressin. Similarly, higher levels of renal nerve stimulation [plus prazosin (100 nM) or phenoxybenzamine (1.0 mg/kg per hr) to block alpha 1-adrenoceptors] did not activate renal alpha 2-adrenoceptors which are associated with the antagonism of the effects of vasopressin. The same level of subthreshold renal nerve stimulation (0.85 +/- 0.14 Hz) in the absence of vasopressin, and without alpha 1- or alpha 2-adrenoceptor blockade, decreased (P less than 0.05) sodium and water excretion. The reversal of this effect by alpha 1-adrenoceptor blockade (prazosin 30 nM) but not alpha 2-adrenoceptor blockade (yohimbine 300 nM) indicates that this effect of renal nerve stimulation is mediated through alpha 1-adrenoceptors. Thus, subthreshold renal nerve stimulation in the rat kidney induces sodium and water retention through activation of alpha 1-adrenoceptors, as shown by others in the rabbit and dog.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990760 TI - The interaction of verapamil and norverapamil with beta-adrenergic receptors. AB - To determine the effect of calcium-channel blockers on beta-adrenergic receptors, we studied the interactions of verapamil, diltiazem, and nifedipine with both human lymphocyte beta 2-adrenergic receptors and rat myocardial beta 1-adrenergic receptors by means of radioligand binding assays. We also determined the functional consequences of these interactions by measuring adenylate cyclase activity. Radioligand binding studies in vitro demonstrated a Ki of verapamil for the lymphocyte beta 2-receptor of 32 +/- 4 microM. Diltiazem and nifedipine were much less potent. In studies of adenylate cyclase activity, verapamil was shown to act as a competitive beta-receptor antagonist. Also, norverapamil, the active metabolite of verapamil, had the highest affinity for the beta-receptor of any of the calcium-channel blockers studied (Ki = 4.2 +/- 0.8 microM). After 1 week of verapamil administration in six normal subjects, isoproterenol-stimulated adenylate cyclase activity in lymphocytes was increased from 60 +/- 4% to 83 +/- 10% over basal activity (p less than .05). This was associated with an increase in lymphocyte beta-receptor affinity for agonist as represented by the decrease in the IC50 for isoproterenol inhibition of [125I] iodocyanopindolol binding from 240 +/- 20 to 170 +/- 10 nM (p less than .05). Additionally, plasma norepinephrine levels were reduced from 206 +/- 58 to 92 +/- 18 pg/ml with 1 week of verapamil treatment (p less than .05). Our data suggest that verapamil affects lymphocyte beta-receptors in vitro and with long-term administration regulates lymphocyte beta-receptor function either directly or indirectly via a reduction in plasma catecholamine levels. PMID- 2990762 TI - Hepatitis A and B markers in commercial quality-control and kit-calibrator sera. PMID- 2990761 TI - Beta 2-microglobulin as a prognostic marker for development of AIDS. PMID- 2990763 TI - ACTH and cortisol responses to ovine corticotrophin-releasing factor in patients with primary and secondary adrenal failure. AB - The ACTH and cortisol responses to an intravenous bolus injection of 100 micrograms ovine CRF were studied in 19 patients with adrenal failure. In all eight patients with primary adrenal failure, plasma ACTH levels increased from a mean basal level of 1494 +/- 431 (SEM) pg/ml to peak value of 2601 +/- 1220 pg/ml at 10 min. In comparison with healthy subjects absolute ACTH increments after ovine CRF were significantly augmented in the patients with Addison's disease (P* less than 0.001), and the absolute ACTH responses after ovine CRF were positively correlated with the basal plasma ACTH levels. The 11 patients with secondary adrenal insufficiency could be subdivided into two groups: (A) those having little or no ACTH and cortisol response to ovine CRF (five patients) and (B) those having prolonged and pronounced ACTH responses with a biphasic pattern and a delayed second peak (six patients), followed in all patients by a marked cortisol increase. These data demonstrate that the CRF-test can discriminate between hypothalamic and pituitary causes of secondary adrenal failure. PMID- 2990764 TI - Isolated adrenocorticotrophin deficiency and empty sella associated with hypothyroidism. AB - A case of isolated ACTH deficiency is described. The patient had an empty sella and longstanding radio-iodine induced hypothyroidism. The association between isolated ACTH deficiency, empty sella and hypothyroidism is reviewed. It has not previously been recognized that the commonest endocrine association of isolated ACTH deficiency is primary thyroid failure. ACTH secretion does not recover following thyroxine replacement treatment. PMID- 2990765 TI - A new family with dexamethasone-suppressible hyperaldosteronism: aldosterone unresponsiveness to angiotensin II. AB - A family with nine siblings in which three siblings have been shown to have dexamethasone-suppressible hyperaldosteronism was studied. All three showed no significant changes of plasma aldosterone during angiotensin II infusion at incremental rates under baseline conditions. After dexamethasone administration (2 mg/d for 4 weeks) plasma renin activity (PRA) rose to normal-supranormal range, while plasma and urinary aldosterone were maintained at low-normal levels. No restoration of aldosterone response to angiotensin II was observed on dexamethasone. Two other siblings were found to be hypertensive with normal baseline data; however, both showed plasma aldosterone hyperresponsiveness to ACTH. In the four normotensive siblings aldosterone response to ACTH was normal. The family pedigree was consistent with autosomal dominant transmission of the disorder. HLA typing showed haplotype A3 Bw35 in all five hypertensive sibs and in one normotensive. In conclusion, low aldosterone compared to PRA, and plasma aldosterone unresponsiveness to angiotensin II infusion before and during dexamethasone, show functional impairment, at least temporary, of the zona glomerulosa. These findings support the hypothesis that aldosterone may be derived from the zona fasciculata in this disorder. PMID- 2990766 TI - Are there genetic determinants for the glucose intolerance of acromegaly? AB - A genetic marker identifying the two parental insulin genes has been studied in 51 Caucasian acromegalics by Southern blot hybridization techniques using a cloned insulin gene probe. Two main DNA insertion classes were detected corresponding to the class 1 and class 3 alleles and thus the following genotypes were found: 1/1, 1/3 and 3/3. The acromegalics were subdivided depending on whether they had a normal (n = 30) or abnormal (n = 21) response to a 50 gm oral glucose tolerance test before treatment. The phenotype frequencies in the former group were 1/1, 43%; 1/3, 53%; and 3/3, 4%; and in the latter group the corresponding figures were 76%, 24% and 0%. The relative incidence of concordance of the phenotype 1/1 with abnormal glucose tolerance in acromegaly was 4.2. This phenotype is also associated with insulin dependent (Type 1) diabetes mellitus. PMID- 2990767 TI - Proopiomelanocortin-derived peptides in the immune system. AB - Cells of the immune system have recently been recognized as a source of the proopiomelanocortin (POMC)-derived peptides, ACTH and endorphins, and their receptors. In this review, the nature of the immunoregulatory and hypothalamic signals which elicit the production of these leukocyte-derived hormones is discussed. Further, comparisons are made between the various proteolytic processing patterns of POMC in leukocytes as compared with those observed in the hypothalamus and anterior or intermediate lobes of the pituitary gland. The possible functioning of leukocyte-derived ACTH and endorphins as well as their pituitary counterparts in a complete regulatory loop between the neuroendocrine and immune systems is described. PMID- 2990768 TI - Observations on a low molecular weight natriuretic and Na-K-ATPase inhibitory material in urine. AB - Natriuretic and Na-K-ATPase inhibitory material prepared from urine by gel filtration on G25 Sephadex was previously found to be of low molecular weight, polar and non-peptide. Although activity appeared to depend on an amino group, tests and radioenzymatic assays for catecholamines suggested that these were not implicated in the natriuretic activity. Further purification of the material included solvent extraction, cation exchange and high performance liquid chromatography. At each stage, fractions were assayed for natriuretic activity, stimulation of G6PD and inhibition of Na-K-ATPase in cytochemical assays, and for digoxin-like activities i.e. inhibition of dog kidney Na-K-ATPase (Sigma), displacement of 3H ouabain bound to cell membranes and cross reaction with antidigoxin antibody. The crude material possessed all activities, but with successive purifications the activities separated from each other and were thus due to different substances. Analyses for catecholamines with HPLC and electrochemical detection revealed that the natriuretic activity was due to dopamine. PMID- 2990769 TI - Plasma Na+-K+ ATPase inhibitory activity in normal and hypertensive subjects: relationship to intracellular electrolytes and blood pressure. AB - The ability of plasma extracts to inhibit Na+-K+ ATPase in vitro (P.I.A.) was tested in 20 normotensives, 10 without (F-) and 10 with (F+) familial hypertension, in 20 borderlines (BL) and in 21 essential hypertensives (EH). In these subjects we also measured intralymphocytic sodium (ILSC) and potassium (ILKC) content, P.R.A. urinary aldosterone and Na+(Na+u), and blood pressure. P.I.A. of EH, BL and F+ subjects was significantly higher than that of F-. 60% of EH and BL and 40% of F+ had P.I.A. values greater than the highest found in F-. P.I.A. was significantly related to mean blood pressure (r = 0.63), to ILSC (r = 0.56), to ILKC (r = -0.56), to ILSC/ILKC ratio (r = 0.71) and to Na+u (r = 0.39) but not to P.R.A. or aldosterone. These data demonstrate that plasma extracts from young subjects prone to hypertension may inhibit sodium pump and that this inhibitor may affect blood pressure by altering the Na+/K+ intracellular ratio. PMID- 2990770 TI - Characteristics of hypertensive patients with increased plasma Na+-K+ pump inhibitory activity. AB - Recent studies in our laboratories and in the laboratories of other investigators suggest the presence of a sodium-potassium pump inhibitor in the plasma of some patients with arterial hypertension. We here review these studies in an attempt to determine what characteristics increase the likelihood of detecting the inhibitor. The review suggests but does not prove that the inhibitor is most likely to be found in males with increased sodium intake, decreased renal function, and decreased plasma renin activity. In future studies of plasma sodium potassium pump inhibitory activity in hypertensive humans, we should pay more attention to the characteristics of the patients. These characteristics include age, sex, race, therapy, stage of hypertension, sodium intake, renal function, and renin status. We should also make an attempt to match the patients properly with normotensive control subjects. PMID- 2990771 TI - The role of endogenous inhibition of Na-K-ATPase in human hypertension--sodium pump activity as a determinant of peripheral vascular resistance. AB - High sodium intake in the presence of an intrinsic or acquired defect in renal sodium excretion will result in extracellular fluid volume (ECFV) expansion which is accompanied by decreased baroreceptor reflex sensitivity. We have shown that ECFV-expansion also stimulates the secretion of an endogenous inhibitor of the Na K-ATPase enzyme and high activity of this sodium transport inhibitor was detected in plasma of patients with primary aldosteronism, the most classical type of volume-dependent hypertension. Thus, vasoconstriction due to inhibition of sodium pump activity of the vascular smooth muscle cell may contribute to the pathogenesis of human arterial hypertension. In analogy, ouabain (8.5 micrograms/kg) when administered i.v. to healthy volunteers inhibited RBC - Na-K ATPase by 49% and significantly increased peripheral vascular resistance by 24 - 36%. The calcium entry blocker nifedipine (10 mg orally) completely prevented ouabain-induced vasoconstriction suggesting that the action of ouabain was mediated by a rise in intracellular calcium. High potassium intake partially abolished the vasoconstrictor effect of ouabain and also significantly increased baroreceptor reflex sensitivity. The results of these studies support the concept that inhibition of the sodium and potassium pump of vascular smooth muscle cells by a yet putative endogenous inhibitor of Na-K-ATPase (natriuretic hormone) may represent a crucial mechanism in the pathogenesis of at least certain forms of essential and secondary hypertension in man. PMID- 2990772 TI - Natriuretic and smooth muscle responses to human urinary natriuretic hormone in rats: relation to blood pressure and kallikrein excretion in patients. AB - Human urine contains a small molecular weight natriuretic substance and similar material isolated from the kidney inhibits Na/K ATPase. Such action on smooth muscle in blood vessels would cause contraction. Human urinary natriuretic material caused contraction of the smooth muscle in the rat anococcygeus muscle and this activity correlated with its natriuretic activity. Known vasoactive substances could not explain the activity of the natriuretic factor when tested on the anococcygeus muscle. The best correlation with blood pressure of the patient was with the log of the ratio of natriuretic activity divided by the kallikrein excretion. A normotensive woman with severe renal failure had very high kallikrein excretion as well as increased natriuretic activity and her data fitted the same correlation as the hypertensives' data. PMID- 2990773 TI - Biochemical studies of rat atrial natriuretic factor. AB - The natriuretic substances were purified from rat atrium (ANF, atrial natriuretic factor) and were shown to be identical with the inhibitor of norepinephrine induced contraction of smooth muscle. Their four native forms were isolated. Amino acid sequence analyses showed they are peptides with 35, 31, 30 and 25 amino acid residues respectively and contain a ring structure consisting of 17 amino acid residues and a disulfide bridge. The presence of a high molecular weight prohormone was shown. cDNA coding for the precursor was cloned and used to deduce the amino acid sequence of the preprohormone. Genomic DNA for ANF was cloned and the presence of two introns were found. Several ANF peptides were synthesized. Structure-function studies showed that the ring structure is essential for the activity. Antibodies produced against the synthetic 25 amino acid residue ANF were used to develop radioimmunoassay. The presence of ANF in rat plasma was demonstrated as evidence that ANF is a circulating hormone. ANF was also found in the hypothalamus of rats. The quantitative determination of the synthetic ability of ANF has been determined by the application of ANF cDNA for the quantification of ANF messenger RNA. Immunohistochemical methods localized ANF in cardiac atriocytes, gonadotrophs in anterior pituitary and adrenal medulla (chromaffin cells). A strong immuno-reactivity was found in dark cells of the collecting ducts of the kidney. ANF increases cyclic GMP in target cells suggesting that cyclic GMP may be the intracellular mediator of ANF action. PMID- 2990774 TI - Pharmacology and receptor binding of atrial natriuretic factor in vascular smooth muscle. AB - Characterization of the vascular pharmacology and receptor binding of atrial natriuretic factor (ANF) has been achieved utilizing a synthetic peptide which contains the sequence and biological activity of ANF. The synthetic ANF (sANF) relaxes aortic segments contracted by agonists or by low (15 to 20 mM) but not high (greater than or equal to 40 mM) concentrations of K+. The relaxation to sANF is well maintained, reversible, independent of the vascular endothelium and correlated with increases in cyclic GMP (with no change in cyclic AMP). Plasma membranes prepared from rabbit aorta and kidney possess high affinity (Kd = 100 pM) specific binding sites for sANF. An excellent correlation exists between the receptor binding and pharmacology for several synthetic analogs of ANF. The presence of these receptors appears consistent with the activation of particulate (but not soluble) guanylate cyclase by sANF. sANF does exhibit a profound regional vasodilator selectivity which can be explained, in part, by changes in receptor density. PMID- 2990775 TI - Renal and systemic effects of atrial natriuretic factor. AB - A survey of the literature reporting the in vivo biological activity of atrial natriuretic factor (ANF) is presented. Both the atrial extracts, in which ANF was first identified, and the peptides which have more recently been synthesized, have been shown to possess diuretic and saluretic properties. In addition, ANF decreased blood pressure and affected levels of aldosterone, renin, vasopressin and guanosine 3',5'-cyclic monophosphate (cGMP). The available evidence concerning likely mechanisms of action of ANF are presented and a pivotal role of ANF is suggested for blood pressure and volume regulation. PMID- 2990777 TI - Laboratory investigation of disorders of the parathyroid glands. PMID- 2990776 TI - Hormone production by tumours: biological and clinical aspects. PMID- 2990778 TI - Diagnosis of human endocrine disorders using recombinant DNA techniques. AB - In this chapter we have reviewed several present and potential examples of DNA studies of hereditary endocrine disorders of humans. For the former, recombinant DNA studies have provided insights into the location and types of molecular derangements underlying these diseases. The gene alterations detected have in turn explained the aetiology of quantitative or qualitative alterations in the hormone product. The same methods used in these studies should be applicable to determining the aetiology of many other genetic disorders that affect these, as well as other, hormones for which specific DNA probes are or will become available. PMID- 2990779 TI - Disorders of the hypothalamic-pituitary-adrenal axis. PMID- 2990781 TI - Serum antibody reacting with placental syncytiotrophoblast in sera of patients with autoimmune diseases--a possible relation to type C RNA retrovirus. AB - This study reports the immunohistological detection of serum antibody reacting with the basal aspect of syncytiotrophoblast of human chorionic villi, where SSAV/GaLV (simian sarcoma associated virus/gibbon ape lymphoma virus) type C retrovirus p30 related antigen was observed by an indirect immunofluorescent method using monospecific antibodies against SSAV p28 and GaLV p29. The immunoglobulin class of this antibody activity called 'the anti-basal aspect of syncytiotrophoblast (anti-BAST)', was exclusively IgM and detected in the sera of both female and male patients with SLE and other autoimmune diseases, but rarely in the sera of normal controls. Immunofluorescent absorption and blocking test revealed that anti-BAST specifically reacted with human placenta and cross reacted with subhuman primate type C RNA retrovirus SSV/SSAV (simian sarcoma virus/simian sarcoma associated virus), but did not cross-react with ATLV (adult T cell leukaemia virus) and BaEV (baboon endogenous retrovirus). These results suggest the presence of a new antigen-antibody system for another human type C retrovirus related antigens(s) and a participation of retrovirus in autoimmune diseases. PMID- 2990780 TI - Elevated numbers of peripheral T cells in inflammatory bowel diseases displaying T9 antigen and Fc alpha receptors. AB - Elevated numbers of peripheral T cells expressing the activation associated antigen T9 are found in patients with active Crohn's disease. Expression of T9 is found to be correlated to the activity of the disease. However the presence of activated peripheral T cells is not restricted to Crohn's disease, but could also be found in other maladies with a supposed involvement of the immune system, e.g. ulcerative colitis, sarcoidosis, connective tissue disease and after organ transplantation. Significant elevation of the number of activated T cells could not be detected in cases of viral or bacterial enteritis and coeliac disease. Analysing the subset of T9 positive T cells with regard to the expression of Fc alpha receptors, a significantly increased number of Fc alpha receptor positive cells, within the subset of T9 positive cells in the peripheral blood of patients with Crohn's disease and ulcerative colitis was found, which could not be demonstrated in the case of other diseases analysed in this study. Thus the T9+ Fc alpha receptor +T cell subset may be considered to be pathognomonic for inflammatory bowel diseases. Analysis of the regulatory properties of T9 positive cells, with regard to the immunoglobulin isotype secretion in a pokeweed mitogen stimulated autologeous B cell assay, suggests that peripheral T9 positive T cells are involved in the suppression of IgA synthesis or secretion. PMID- 2990783 TI - T lymphomas associated with human T-cell leukemia-lymphoma virus may show phenotypic and functional differences from adult T-cell leukemias. AB - We report phenotypic and functional characterization of lymph node cells from 27 patients with T lymphoma from the Nagasaki district of Japan. Nagasaki is one of several areas where adult T-cell leukemia is endemic, and sera from 18 of 23 patients examined (78.3%) were positive for antibody to adult T-cell leukemia associated antigen. The majority of cases (24 of 27) showed the Leu-2a-, 3a+, MASO36c- phenotype similar to adult T-cell leukemia, but seemed to be subdivided into four groups according to the presence or absence of Leu-1 antigen and the receptor for sheep erythrocytes (SRBC). Eleven cases had both Leu-1 antigen and the receptor for SRBC (E-RFC+, Leu-1+, 2a-, 3a+), but 6 cases lacked Leu-1 antigen (E-RFC+, Leu-1-, 2a-, 3a+), 4 cases lacked receptors for SRBC (E-RFC-, Leu-1+, 2a-, 3a+), and 3 cases lacked both of these markers (E-RFC-, Leu-1-, 2a-, 3a+). The effects of lymph node T cells on pokeweed mitogen (PWM)-induced normal B-cell differentiation was studied in 10 cases. Lymph node T cells of 5 cases showed helper activity without any suppressor activity, and 2 cases showed suppressor activity with almost no detectable helper activity, similar to adult T cell leukemia cells. Two other cases lacked particular functional effects. These results suggest the possibility that Leu-3a+ T-cell lymphoma may be functionally subdivided into at least two types, a helper type and a suppressor (adult T-cell leukemia) type. PMID- 2990782 TI - Detection of IgA heavy chain constant region genes in IgA deficient donors: evidence against gene deletions. AB - Sixty-six donors with selective IgA deficiency and one patient with selective IgA2 deficiency were investigated for immunoglobulin gene defects using restriction enzyme digestions and Southern blot analysis. All patients carried alpha 1 and alpha 2 genes in their genome, suggesting that large deletions are uncommon causes for IgA deficiency. Digestion with Bam HI, Pst I and Pvu II, did not reveal any polymorphism in the studied samples. PMID- 2990784 TI - Characteristics of chondrocyte responses to a human interleukin 1-like factor. AB - The objective of these studies was to characterize some aspects of collagenase production by rabbit articular chondrocytes cultured with stimulated monocyte supernatants. Supernatants from human monocytes stimulated with 20 ng/ml bacterial lipopolysaccharide (LPS) induced the synthesis and secretion of latent collagenase by the chondrocytes beginning at 6 hr. The time course and dose response of collagenase production by the chondrocytes were identical using crude monocyte supernatants or semipurified interleukin 1 (IL-1). Recombinant or purified human interleukin 2 failed to induce collagenase production in the cultured chondrocytes. The response of the chondrocytes was inhibited by actinomycin D or cycloheximide and not by corticosteroids. Phorbol myristate acetate (PMA) alone failed to directly stimulate the chondrocytes. However, PMA led to enhanced collagenase production by chondrocytes when incubated with submaximal amounts of LPS-stimulated monocyte supernatant or semipurified IL-1. LPS alone in amounts between 0.1 and 10.0 micrograms/ml directly stimulated collagenase production in chondrocytes between 4 and 11 days in culture. These data confirm those of other laboratories that IL-1 may be the active factor in monocyte supernatants responsible for inducing collagenase production in cultured chondrocytes. Further characterization of this response indicates that the collagenase is not preformed in the cells and stimulation of its production is not inhibited by corticosteroids. Cell supernatants or IL-1 preparations containing PMA as low as 1.0 ng/ml or LPS as low as 1.0 microgram/ml may give falsely high values for IL-1 activity when assayed by stimulation of collagenase production in cultured chondrocytes. PMID- 2990785 TI - Pelvic inflammatory disease--use of appropriate antibiotics. PMID- 2990786 TI - Familial amyloid polyneuropathy: report of a family. AB - We report an Italian family affected by familial amyloid polyneuropathy. In two siblings, the diagnosis was confirmed by skin biopsy which revealed an unusual involvement of cutaneous nerves. Immunohistochemical studies confirmed the presence of prealbumin in the amyloid deposits. PMID- 2990787 TI - Primitive neuroectodermal tumor with choroid plexus differentiation. AB - A case of a cerebral juvenile undifferentiated round cell tumor with unusual formation of glandular structures is presented. The round cell component showed focal expression of glial fibrillary acidic protein, S-100 protein, neurofilament protein and neuron-specific enolase. The glandular part consisted of papillary epithelial structures with a central PAS-positive lumen. Ultrastructural findings supported the interpretation of these parts as choroid plexus. This route of differentiation was recognizable at the first operation at the age of 5 years and was much more prominent in the recurrence found 7 1/2 years later. The unusually benign course of this primitive neuroectodermal tumor (PNET) may be due to its intrinsic progressive tendency towards choroid plexus differentiation. PMID- 2990788 TI - Peripheral neuropathy and paraproteinemia: an immunohistochemical and serologic study. AB - Four of 132 consequtive patients with paraproteinemia and with an M-component presented exclusively with neurological symptoms. A sural nerve biopsy was performed in all patients. In one case direct immunofluorescence revealed deposition of immunoglobulin corresponding to the M-component along the perineurium. No deposition of immunoglobulin was found along the myelin sheaths and the patient's serum revealed no immunoreactivity towards normal peripheral myelin or other components of normal peripheral nerves. In a second case, deposition of immunoglobulin corresponding to the M-component was found along the myelin sheaths, and this patient's paraprotein showed immunoreactivity towards normal peripheral myelin. Two patients showed no in situ bound immunoglobulin in the nerve biopsies and their sera did not react with any component in normal peripheral nerves. The patient with deposition of immunoglobulin along the perineurium had severe demyelination and severe peripheral sensory-motor neuropathy, while the patient with immunoglobulin deposition along the myelin sheaths had a slight sensory neuropathy which had not progressed for 4 years. Based on these findings the pathogenesis of neuropathy in patients with paraproteinemia is discussed, and it is concluded that although there is evidence for immune-mediated neuropathy in the patients, the histologic, immunohistochemical and clinical data are conflicting and the exact pathogenic pathway is still unknown. PMID- 2990789 TI - The free synovial graft as a shield for collagen ingrowth in cruciate ligament repair. AB - The anterior cruciate ligament (ACL) was excised bilaterally in the knee joint of three dogs and was replaced in a conventional manner with a composite polyglycolic acid (PGA) and Dacron braided ligament prosthesis. In one knee of each dog, the ligaments were wrapped with a free synovial graft taken from the suprapatellar pouch in an attempt to keep the prosthesis extrasynovial, while the ligaments in the opposite three knees were left uncovered and thus directly exposed to synovial fluid. The dogs were killed at approximately one year postoperation, and microscopic sections of all ligaments were taken. The three knees with the synovial sheath showed a significantly greater bulk of fibrous tissue along the ligament scaffold grossly and a greater abundance of collagen producing cells microscopically, compared to the unwrapped ligaments. The gross and microscopic findings were identical in each of the three knees with the covered ligaments and were equally consistent in the three that had been left unprotected from the synovial fluid. These findings suggest that a free synovial graft may simulate the normal protective role of the synovial membrane and allow and/or induce collagen ingrowth in synthetic or autogeneic tissue stints for the repair of acute cruciate ligament tears. PMID- 2990790 TI - [Congenital dermal sinus with lesions in the conus medullaris and the cauda equina--with a case report]. PMID- 2990791 TI - [An autopsy case of carcinomatous sensory neuropathy. The correlation between the morphometric findings of the primary sensory neuron and clinical findings]. PMID- 2990792 TI - [Visuomotor ataxia. Clinical and CT scan studies in three cases]. PMID- 2990793 TI - Variant flow patterns in radionuclide cerebral imaging performed for brain death. AB - Radionuclide cerebral imaging (RCI) is a commonly used technique for detection of intracranial blood flow in patients with clinically suspected brain death. In 115 studies performed on 95 patients in a six-year period at the Geisinger Medical Center, a number of variant flow patterns have been observed. Although all have been described previously, there has been no review of these patterns on studies performed for brain death. The significance of normal blood flow, absent blood flow, isolated superior sagittal sinus activity, luxury perfusion, hypoperfusion, hemiperfusion, and jugular venous reflux is discussed. Any demonstrable intracranial blood pool activity is presently considered to counter a diagnosis of brain death, except that jugular reflux renders an examination uninterpretable. Technique must be top quality but, even so, RCI cannot evaluate basilar brain flow. Nonetheless, RCI is a valuable adjunct in helping to establish the clinical diagnosis of brain death. PMID- 2990794 TI - Imaging of an actively bleeding aortoduodenal fistula. AB - Diagnosis of gastrointestinal bleeding remains a difficult problem. Radionuclides have been used widely to detect active GI bleeding. A case of actively bleeding aortoduodenal fistula detected on radionuclide imaging is presented. PMID- 2990795 TI - Nonrotation of bowel. Possible pitfalls in Meckel's scanning. PMID- 2990796 TI - Indium-111 chloride for detecting suspected hepatomas in patients with focal defects on technetium-99m sulfur colloid liver imaging. AB - Twenty-three patients with hepatic cirrhosis and focal defects on Tc-99m sulfur colloid (SC) scintigrams were restudied with In-111 chloride to determine if indium localization in the focal defect is indicative of a hepatoma. Seven of eight patients with proven hepatomas had positive studies; however, six of 15 patients without hepatomas also had studies interpreted as positive. Thus, In-111 chloride is highly sensitive for the detection of hepatomas, and a negative indium study would militate against this diagnosis. The high false-positive rate found may be due to technical factors rather than a lack of specificity of localization; the experience of others seems to support this impression. At present, In-111 chloride scintigraphy for focal hepatic defects appears to be useful in ruling out hepatoma. PMID- 2990797 TI - Hydrops fetalis with supraventricular tachycardia and cytomegalovirus infection. PMID- 2990798 TI - Enalaprilat, an intravenous angiotensin-converting enzyme inhibitor, in hypertensive crises. AB - The effect of enalaprilat (MK-422), a newly synthesized, intravenous, nonsulfhydryl, angiotensin-converting enzyme inhibitor, was studied in seven patients with either severe or malignant hypertension. All subjects initially received a 1 mg bolus injection of enalaprilat followed in 30 minutes by 10 mg. Five subjects received an additional 40 mg. Mean (+/- SE) pretreatment blood pressure for the group was 226 +/- 9/141 +/- 7 mm Hg. Five minutes after the 1 mg enalaprilat dose, blood pressure decreased to 211 +/- 10/131 +/- 9 mm Hg and further fell to 201 +/- 14/123 +/- 11 mm Hg at 30 minutes. The maximal reduction in blood pressure to 169 +/- 14/112 +/- 10 mm Hg occurred 30 minutes after the 10 mg dose. No further blood pressure reduction was observed in those subjects who received the additional 40 mg dose. Within the entire group, five subjects exhibited sustained blood pressure reduction. No adverse side effects or symptomatic hypotension occurred in any subject. PMID- 2990799 TI - The value of angiotensin-I-converting enzyme determinations in malignant and other diseases. AB - Mean values for serum angiotensin-I-converting enzyme (SACE), determined spectrophotometrically in 648 subjects, using the synthetic substrate hippuryl-L histidyl-L-leucine, and expressed in units per milliliter, were: controls, 11.11 +/- 3.97 (n = 89); lung cancer, 6.50 +/- 3.26 (n = 87); tuberculosis of the lung, 8.93 +/- 4.60 (n = 68); pulmonary sarcoidosis, 21.18 +/- 14.93 (n = 48); pneumonia, 9.81 +/- 6.83 (n = 52); fibrosis, 11.18 +/- 8.26 (n = 34); diabetes mellitus, 10.90 +/- 7.51 (n = 29); ischemic heart disease, 8.98 +/- 6.19 (n = 42); pulmonary embolism, 13.20 +/- 3.91 (n = 5); and lymphomas, 11.66 +/- 5.44 (n = 36). The lowest values for SACE (5.92 +/- 1.95) were observed in 7 patients with pulmonary metastases. No relationship could be found between SACE and other laboratory parameters, nor between the enzyme activity in men and women. Evidence suggests that low SACE activity is often associated with extrapulmonary cancers of various organs. Levels were significantly decreased in cancer of the lung and pulmonary metastases and significantly (p less than 0.001) increased in sarcoidosis compared with other diseases, suggesting that SACE activity may be of value in the diagnosis and prognosis of cancer of the lung. PMID- 2990800 TI - In vivo accumulation and effects of mercuric chloride on tissue phosphatases of Notopterus notopterus. AB - Sublethal concentrations of mercuric chloride inhibited the liver, kidney and gill enzymes, acid, alkaline, and glucose-6-phosphatase of the fish Notopterus notopterus, which had been exposed to the heavy metal for 30 days. Higher concentrations produced highly significant (p less than 0.001) inhibition, whereas concentrations that were 1/25th the lethal concentration caused significant (p less than 0.05)/insignificant inhibition. The quantity of mercuric chloride accumulated in tissues correlated with the degree of enzyme inhibition. PMID- 2990801 TI - Alteration of cellular calcium metabolism as primary cause of hypertension. AB - In the pathogenesis of hypertension, the importance of intracellular calcium is increasing. Clinical and experimental studies of essential hypertension indicate a pathological increase of intracellular Ca2+ in this disease. In the past, changes in cellular Na+ and its transport mechanisms were considered the triggering factors and Na+-Ca2+ exchange was attributed a decisive influence on intracellular homeostasis. Recently, a reduced Ca2+-binding capacity of the cellular membrane was observed in hypertension, which could have been due to a defect of the Ca2+-ATPase or its control. It is therefore necessary to establish the hypothesis that changes in the cellular Ca2+ metabolism associated with an increase in the intracellular Ca2+ concentration may be the primary cause of hypertension. Disorders of Na+ transport can also be traced to the increase in intracellular Ca2+ and were thus a consequence but not the cause of the increased intracellular Ca2+ concentration. PMID- 2990802 TI - [Hepatic encephalopathy. Pathogenetic evaluation and current possibilities of treatment]. PMID- 2990803 TI - Conspectus. Corticotropin-releasing factor. PMID- 2990804 TI - Communicable disease report. October to December 1984. PMID- 2990805 TI - Properties of avian, bovine and porcine erythrocyte membranes. AB - Bovine, porcine and avian EMP were isolated and compared for some physical and chemical properties. Some differences in the compositions of three EMPs were observed. The avian EMP contained less carbohydrate than the bovine and porcine EMPs. Some differences in the monosaccharide distributions for the three preparations were revealed. The profiles obtained by SDS-polyacrylamide gel electrophoresis of the preparation indicated a complex (and different for each preparation) nature of the component polypeptides and glycopeptides. PMID- 2990806 TI - Evidence for existence of a cytosol 5'-nucleotidase in chicken heart: comparison of some properties of heart and liver enzymes. AB - A 5'-nucleotidase was purified from chicken heart. Kinetic properties of the enzyme were similar to the cytosol 5'-nucleotidase previously reported for chicken liver and rat liver. This strongly suggests the existence of the same type of the cytosol 5'-nucleotidase in cardiac tissue that has been reported for hepatic tissue of various animals. PMID- 2990807 TI - Metabolism in marine flatfish. II. Protein digestion in Dover sole (Solea solea L.). AB - The digestive tracts of adult and juvenile Dover sole were examined for protease activities. A pepsin-like protease with an optimal pH value of 1.7 predominated in the stomach region, but the main endoprotease action in the foregut, midgut and hindgut regions was optimal in the range of pH 9.5-10.5 and showed good activity towards elastin orcein. Experiments using synthetic substrates suggested the presence of chymotrypsin- and trypsin-like activities optimal between pH 7 and 8. Collagenase activity was also shown to exist in this pH region. The presence of enzymes corresponding to carboxypeptidases a and b and leucine aminopeptidase was indicated. The possible significance of these results to the farming of Dover sole is discussed. PMID- 2990808 TI - Influence of salinity and ratio of lipid to protein in diets on certain enzyme activities in rainbow trout (Salmo gairdneri Richardson). AB - The connection between metabolic and sea water adaptation of the rainbow trout was investigated. The rainbow trout were kept in fresh water and diluted sea water of 8 and 20 0/00 S at 16 degrees C and fed on three different diets for 51 days. Hyperosmotic salinity (20 0/00) tends to inhibit growth in rainbow trout by reducing the food conversion efficiency. A higher protein concentration in the diet can partly compensate for this effect. The liver IDH, G6PDH and 6PGDH activities of the rainbow trout are influenced only by food quality, whereas the liver G1DH, AspT and A1T activities, like the muscle A1T, are also affected by salinity. The salinity had no significant effect on the activities of the kidney enzymes we investigated (Na/K-ATPase, G1DH, A1T, AspT) or of the muscle AspT in these experiments. PMID- 2990809 TI - Reduction and oxidation of cytochrome C by Hymenolepis diminuta (Cestoda) mitochondria. AB - Mitochondrial membranes of adult Hymenolepis diminuta catalyzed inhibitor sensitive ferricytochrome c reduction. Cytochrome c reductase activity was noted when NAD(P)H or succinate served as the reductant with the NADH-coupled reaction being most prominent. Both rotenone-sensitive and -insensitive reduced pyridine nucleotide-coupled activities were apparent. Ferrocytochrome c oxidase activity also was catalyzed by H. diminuta mitochondrial membranes and this reaction was sensitive to azide and cyanide. A cytochrome c peroxidase activity was associated primarily with the mitochondrial soluble fraction of adult H. diminuta. The possibility that the activities observed may contribute to the elimination of peroxide in the helminth system is considered. PMID- 2990810 TI - Glycolytic enzyme levels in synaptosomes. AB - The specific activities of glucosephosphate isomerase, aldolase, triosephosphate isomerase, glyceraldehydephosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, pyruvate kinase and lactate dehydrogenase were all higher in the synaptoplasmic fraction from rat brain than in 100,000 g supernatant fraction of rat brain homogenates when the supernatants were prepared in high ionic strength solutions. Four enzymes in synaptosomes and two enzymes in homogenates were associated with particulate fractions as indicated by the large increase in specific activity of the enzymes when samples were treated with 0.3 M KCl before centrifugation. Glucosephosphate isomerase, aldolase, pyruvate kinase and lactate dehydrogenase were the enzymes that showed a large increase in specific activity following salt treatment of isolated, synaptosomal membrane while aldolase and pyruvate kinase were the two enzymes which showed a large increase in specific activity in the high speed supernatant fractions. Because the specific activities of many enzymes are found to be elevated not only in synaptosomes but in synaptosomal membrane fractions it is suggested that these enzymes may provide the potential for significantly enhanced glycolysis at these locations. PMID- 2990811 TI - Distribution of enzymes of purine metabolism in lymphocytes of horse, Equus caballus. AB - A microassay requiring as few as 2 X 10(5) cells per assay was developed for systematic analysis of 9 purine enzymes in lymphocytes from equine peripheral blood, spleen, lymph node, thymus and bone marrow. The activities of adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), adenosine kinase (AK), deoxyadenosine kinase (dAK), deoxycytidine kinase (dCK), 5'-nucleotidase (5'-N), AMP deaminase, hypoxanthine-guanine phosphoribosyl transferase (HGPRT or HPRT), and adenine phosphoribosyl transferase (APRT) were measured by this microassay in lymphocytes from peripheral blood from four different breeds of horses (Arabian, Quarter Horse, Thoroughbred and Shetland Pony). There were no significant differences in the enzyme activities among the various breeds. Peripheral blood lymphocytes (PBL) from foals exhibited enzyme activities similar to those observed for adult animals. All lymphoid tissue contained similar levels of activity for each kinase (AK, dAK and dCK). Spleen had the highest activity for ADA, PNP, 5'-N, and HGPRT. The lowest activity for ADA, APRT, PNP and AMP deaminase was found in thymus. Enzymatic activities that varied the most among the tissue were 5'-N, ADA, APRT, HGPRT and AMP deaminase. PMID- 2990812 TI - Enzymatic activities in slow and fast denervated old rat muscles. AB - The activities of five enzymes have been studied quantitatively in denervated extensor digitorum longus, gastrocnemius and soleus muscles of 24-month-old rats. The results have been compared with those obtained from normal muscles of a similar age group of rats. Three weeks after denervation, the activity of hexokinase was increased in gastrocnemius and extensor digitorum longus. Phosphofructokinase, lactate dehydrogenase, malate dehydrogenase and 3 hydroxyacyl-CoA-dehydrogenase showed decreased activities. These results suggest that enzyme which represents glucose uptake increased its activity in fast muscles and that enzymes for anaerobic glycolysis, lactate fermentation, citric acid cycle and beta-oxidation had a decreased activity in slow and fast muscles. PMID- 2990813 TI - New observations of soft tissue sarcomas with contrast medium-enhanced computed tomography. AB - Computed tomography examinations were performed on 74 patients who presented to the Children's Hospital of Philadelphia and the Hospital of the University of Pennsylvania with a known or suspected diagnosis of primary or secondary soft tissue sarcoma. Focal masses were detected on computed tomography study in 59 patients. These masses were classified into three broad categories: centrally necrotic masses with a large predominantly liquefactive center and higher density periphery (29); multilocular, septated masses with distinct linear bands or striations (21); and miscellaneous masses (9). The miscellaneous category included six inhomogeneous and three homogeneous masses. The apparent density differences within these sarcomas were best appreciated on dynamic postcontrast scans. The computed tomography appearance of these sarcomas may be explained by the pathologic findings of cystic degeneration, extensive necrosis, central cavitation, focal hemorrhage, and myxoid changes. PMID- 2990814 TI - Nuclear magnetic resonance imaging of spinal dissemination of intracranial germ cell tumor: a case report. AB - The authors report a case of multiple spinal dissemination originating from an intracranial germ cell tumor. Nuclear magnetic resonance imaging is a useful and noninvasive study for evaluating multiple dissemination and the effects of therapy, whereas myelography is invasive and demonstrates the lesion as a negative, indirect shadow defect. The usefulness and some problems of nuclear magnetic resonance imaging of spinal tumor are discussed. PMID- 2990815 TI - Intracranial perineural metastasis of adenoid cystic carcinoma of head and neck. AB - Three cases of intracranial perineural metastasis from adenoid cystic carcinoma of the head and neck are presented. The metastases presented more than 1 year (14 months to 5.5 years) after the primary tumor had been successfully treated without any evidence of local recurrence. Computed tomography of the intracranial metastasis demonstrated well-demarcated, hyperdense, enhancing extraaxial masses indistinguishable from meningioma. An intracranial extraaxial mass lesion with the appearance of a meningioma in a patient with known adenoid cystic carcinoma of the head and neck, regardless of the presence or absence of local recurrence, should be considered as metastasis until proved otherwise. PMID- 2990816 TI - Intraperitoneal hepatocellular carcinomatosis. AB - The sonographic and computed tomographic features of liver involvement with hepatocellular carcinoma are well known. The extrahepatic manifestations of hepatocellular carcinoma are not uncommon, but have received little attention in the radiologic literature. We present a case in which intraperitoneal carcinomatosis and portal vein thrombosis were the dominant findings on sonography and computed tomography. PMID- 2990818 TI - Roles of cyclic AMP and Ca in epithelial ion transport across corneal epithelium: a review. AB - The messenger roles of cyclic AMP and the calcium ion in stimulus-secretion coupling are considered in the frog and bovine corneal epithelium, respectively. In the frog cornea, epinephrine stimulates net C1 transport by increasing cyclic AMP content. This stimulation is associated with a larger apical membrane C1 conductance and basolateral membrane ionic conductance. The response of the apical membrane conductance is thought to result from an increase in cyclic AMP content whereas the basolateral membrane ionic conductance increase is unrelated based on measurements of the effects of the calcium channel antagonist, diltiazem, and the beta agonist, isoproterenol, on the electrical parameters and cyclic AMP content. The basolateral membrane is essentially K permselective since the K channel blocker, Ba, depolarized the intracellular potential difference and increased the basolateral membrane resistance. Diltiazem had even larger effects on these parameters suggesting that this compound is a more effective inhibitor of K channel activity than barium. In broken cell preparations of bovine corneal epithelium, a high affinity form of Ca + Mg activated ATPase is present (Km = .06 microM for Ca) and is essentially of plasma membrane origin. This ATPase activation is at a Ca activity similar to the expected intracellular value and suggests that this activity is the enzymatic basis for net Ca transport. PMID- 2990817 TI - Isolation and evaluation of target cells in glaucoma research: hormone receptors and drug responses. AB - To investigate the actions of drugs and hormones which influence intraocular pressure (IOP), we have been measuring receptor binding characteristics and dose response effects using individual target cells from the anterior segment. Similar methods have also permitted us to develop sensitive radio-receptor assays for glucocorticoids and beta adrenergic antagonists. These methods provide a means to obtain new and potentially useful information regarding the efficacy and specificity of ocular drug therapy. PMID- 2990819 TI - Anion sensitive ATPase in human cornea. AB - In human cornea an anion sensitive ATPase is present. The highest specific activity was found in the endothelium, whereas the epithelium contained the highest total activity. The enzyme was stimulated by bicarbonate and sulfite and inhibited by thiocyanate. the majority of the enzyme was localized in the mitochondria but some activity was also detected in the plasma membranes. Atractyloside inhibited only the mitochondrial anion ATPase. PMID- 2990820 TI - Effect of temperature on the rainbow trout lens. AB - Membrane response to the various temperatures as one of the external factors was investigated in the lenses of the poikilothermal animal and the homothermal animal. The rainbow trout lens was used as the poikilothermal material and the rat lens as the homothermal material. The rainbow trout lens maintained in vitro its transparency without the changes of cation balance at 0 degrees C - 25 degrees C, while cold cataract developed in the rat lens under the same conditions. Na, K-ATPase activity was detected at 0 degrees C in rainbow trout lens but it was not detected in the rat lens. Lactic acid in the rainbow trout lens was produced for 30 successive days at 0 degrees C, while that in the rat lens was not produced. The cataract developed at 37 degrees C in rainbow trout lens, which we called "warm cataract." Warm cataract developed not only when the lens was incubated in vitro but also when rainbow trout was kept in fresh water at 37 degrees C. Significant differences were detected in components of membrane lipids in the rainbow trout lenses compared to bovine lenses as the mammalian lens. The cholesterol/phospholipid ratio in the trout lens membrane was lower than that in bovine lens. This suggests that a poikilothermal animal lens can maintain the membrane fluidity at low temperatures. These results might suggest that the membrane characteristics in the rainbow trout lens play a role to maintain its transparency at low temperatures. PMID- 2990821 TI - Cytotoxic effects of internal calcium on lens physiology: a review. AB - While calcium is possibly involved in cataractogenesis, it is unquestionably involved in normal lens physiology. Numerous reports have documented the many cellular processes in other tissues affected by alterations in cellular levels of calcium. The homeostasis of the lens is no less dependent on the critical balance of intracellular calcium. With advances being made in calcium-sensitive microelectrodes and pioneering studies progressing in ion channel electrophysiology, interest in calcium metabolism in the lens has been intensified. This report is an attempt to review recent findings that deal solely with biochemical changes resulting from calcium imbalances in the lens interior. PMID- 2990822 TI - Stimulus-secretion coupling in exocrine glands: the role of inositol-1,4,5 trisphosphate, calcium and cAMP. AB - Enzyme, electrolyte and fluid secretion from exocrine glands is stimulated by neurotransmitters and peptide hormones. Whereas for some of these secretagogues calcium is an important intracellular messenger, for others it is cyclic AMP. Regulation of steady state free Ca2+ concentration at rest and at stimulation have been studied in isolated permeabilized acinar cells from pancreas, parotid and lacrimal glands by measuring the free Ca2+ concentration of the surrounding incubation medium with a Ca2+-specific macroelectrode. Ca2+ transport mechanisms have been further characterized in subcellular membrane fractions by measuring 45Ca2+ uptake into membrane vesicles from rough endoplasmic reticulum (RER) and plasma membranes (PM). The data show that the intracellular messenger for secretagogue-induced Ca2+ release from RER is inositol-1,4,5-trisphosphate (IP3) which is produced during stimulation by phospholipase C mediated hydrolysis of phosphatidylinositol-bisphosphate. At rest both Ca2+ uptake into RER and Ca2+ extrusion from the cell is promoted by (Ca2+ + Mg2+)-ATPases with different characteristics in both types of membranes and by a coupled Na+/Ca2+ countertransport in the PM which keep cytosolic free Ca2+ concentration at a low level of approximately 2 - 4 X 10(-7) mol/l. During stimulation the Ca2+ permeability of endoplasmic reticulum membrane increases via IP3 and that of the PM by a yet unknown "receptor-operated" mechanism. These events lead to increase in cytosolic free Ca2+ concentration that is a trigger for enzyme, electrolyte and fluid secretion. PMID- 2990823 TI - Left ventricular dilatation. Prognostic value in severe left ventricular dysfunction secondary to coronary artery disease. AB - This study determines the noninvasive prognostic predictors (using radionuclide angiography) in patients with severe left ventricular dysfunction (resting ejection fraction less than or equal to 35 percent) secondary to coronary artery disease. We retrospectively evaluated 94 such patients using rest and exercise radionuclide ventriculography. At a mean follow-up of 16 months, cardiac events occurred in 22 patients: ten patients died of cardiac causes, five patients sustained nonfatal myocardial infarction, and seven patients developed severe congestive heart failure (class 4). Results indicate that patients with severe left ventricular dysfunction may be stratified into different risk groups according to left ventricular size. Marked left ventricular dilatation identifies a subgroup at high risk. PMID- 2990825 TI - Hepatectomy for primary liver cancer in 114 cases. A follow-up study with long term results. PMID- 2990824 TI - A presumptive role for leukotrienes in obstructive airways diseases. AB - The likelihood that the leukotriene products derived from the 5-lipoxygenase pathway mediate aspects of obstructive airways diseases is strongly suggested by their documented capacities to effect airway spasmogenicity, airway hyperreactivity, tissue edema formation, mucus secretion, and tissue infiltration by leukocytes. That the various leukotriene components of SRS-A have unique receptors on responding tissues and are recoverable from airway surfaces in several inflammatory lung diseases and that several resident and infiltrating cell types have significant potential for leukotriene biosynthesis lend further support to their postulated pathobiologic roles. To fulfill Koch's postulates for proof of leukotrienes' etiologic role, it remains to be shown that inhibition of their biosynthesis or specific antagonism at their end-organ receptors can greatly ameliorate these disease states. PMID- 2990826 TI - [Surgical abdomen in the acquired immunodeficiency syndrome. Apropos of 4 cases]. PMID- 2990827 TI - [Vaginal metastasis of choriocarcinoma and invasive mole: clinical and pathological characteristics and treatment]. PMID- 2990829 TI - [Clinico-pathologic study of 184 cases of minor salivary gland tumor]. PMID- 2990828 TI - Organization and chromosomal specificity of autosomal homologs of human Y chromosome repeated DNA. AB - The human Y chromosome contains a group of repeated DNA elements, identified as 3.4-kilobase pair (kb) fragments in Hae III digests of male genomic DNA, which contain both Y-specific and non-Y-specific sequences. We have used these 3.4-kb Hae III Y fragments to explore the organizational properties and chromosomal distribution of the autosomal homologs of the non-Y-specific (NYS) 3.4-kb Hae III Y elements. Three distinct organizations, termed domains, have been identified and shown to have major concentrations on separate chromosomes. We have established that domain K is located on chromosome 15 and domain D on chromosome 16 and suggested that domain R is on chromosome 1. Our findings suggest that each domain is composed of a tandemly arrayed cluster of a regularly repeating unit containing two sets of repeated sequences: one that is homologous to the NYS 3.4 kb Hae III Y sequences and one that does not cross-react with the 3.4-kb Hae III Y repeats. Thus, these autosomal repeated DNA domains, like their Y chromosome counterparts, consist of a complex mixture of repeated DNA elements interspersed among each other in ways that lead to defined periodicities. Although each of the three identified autosomal domains cross-reacts with 3.4-kb Hae III Y fragments purified from genomic DNA, the length periodicities and sequence content of the autosomal domains are chromosome specific. The organizational properties and chromosomal distribution of these NYS 3.4-kb Hae III homologs seem inconsistent with stochastic mechanisms of sequence diffusion between chromosomes. PMID- 2990830 TI - [Virological study on 7 cases of atypical poliomyelitis]. PMID- 2990831 TI - [Effects of certain inflammatory mediators on the development of pulmonary edema induced by steam inhalation]. PMID- 2990832 TI - [Clinical application of renal artery embolization analysis of 40 cases]. PMID- 2990833 TI - [Use of a new phosphate glass composite material as artificial bone]. PMID- 2990834 TI - Flow cytometric evaluation of cell dispersion from human head and neck tumors. AB - The preparation of single-cell suspensions from 25 human head and neck tumors is described. Dispersal was performed overnight at 4 degrees C under slight agitation of the tissue suspensions using various combinations of enzymes and additives. The cell suspensions were examined for number of cells released, viability, amount of debris, and DNA distribution by means of flow cytometry (FCM). It was shown that both trypsin/dithioerythritol (TD) and collagenase/D Nase (CDse) were of value in dispersing single cells from tumor tissue. In contrast to CDse, incubation with TD appeared to be cytolytic to normal lymphocytes. In a number of cases, DNA-FCM revealed ploidy abnormalities in a TD suspension, which were not discernible in the concurrent CDse-suspension. Cell culture of primary cell suspensions corroborated the reliability of the DNA-FCM measurements. Pretreatment with CDse improved tumor disaggregation by TD and indicated a different dispersal capacity. Addition of Ca2+ and Mg2+ ions to the dispersal mixtures and preincubation of tumor slices in complete medium for 1 day before initiation of cell dispersion influenced favorably the quality of the cell suspension. PMID- 2990836 TI - Small-cell carcinoma of the rectum. AB - Small-cell carcinoma of the rectum is a rare tumor. We treated a patient with small-cell carcinoma of the rectum with radiation and multidrug regimen being used for small-cell carcinoma of the lung. Within two months of chemotherapy, the primary lesion, as evaluated by rectoscopy, biopsy, and CT scan, had resolved completely. The patient was in complete remission for 12 months after initiation of chemotherapy and died of widespread metastases. At autopsy, no residual tumor was found in the rectum. This case stresses the importance of ultrastructural study in the differential diagnosis of small-cell cancer of the rectum and the fact that this tumor can be treated in the same fashion as for small-cell carcinoma of the lung with multidrug chemotherapy and radiation therapy to achieve local control. PMID- 2990835 TI - DNA distribution in non-small-cell lung carcinomas and its relationship to clinical behavior. AB - A study of 187 surgical specimens of tumors of patients with non-small-cell lung carcinomas was carried out by means of flow cytometry. Eighty-four percent of the tumors were classified as tumors with abnormal DNA stemlines (DNA aneuploidy). Patients with tumors demonstrating DNA aneuploidy had significantly shorter survival times than those with tumors demonstrating DNA diploidy (p = .009). Cell cycle analysis was possible in 122 tumors. Patients whose tumors had 0-8% S-phase cells died later than patients whose tumors had 9-16% S-phase cells (p = .018). In addition, patients with tumors with a low fraction of labeled S-phase cells (autoradiography) had a better prognosis than patients with tumors with a high proportion of labeled S-phase cells (p = .041). PMID- 2990838 TI - [Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels in cancer tissue, plasma and urine of esophageal cancer patients]. PMID- 2990837 TI - Reflex adrenergic inhibition of colonic motility in anesthetized rat caused by nociceptive stimuli of peritoneum. An alpha 2-adrenoceptor-mediated response. AB - Nociceptive stimulation of the peritoneum inhibits colonic motility via a sympathoadrenergic reflex. This sympathetic reflex most probably induces the inhibition indirectly via a presynaptic blockade of cholinergic transmission. To further clarify this reflex, colonic motility was continuously recorded by a volume method in rats exposed to nociceptive abdominal stimulation and intravenous drug administration. Intraabdominally applied HCl inhibited colonic motility, which was unaffected by nonselective beta-adrenoceptor blockade (propranolol). However, the nonselective alpha-adrenoceptor antagonist phentolamine and the selective alpha 2-adrenoceptor antagonist yohimbine restored motility. Spontaneous colonic motility was unaffected by the nonselective beta adrenoceptor agonist (isoproterenol) and the selective alpha 1-adrenoceptor agonist (L-phenylephrine). On the other hand, a nonselective alpha-agonist (noradrenaline) and a selective alpha 2-adrenoceptor agonist (clonidine) inhibited spontaneous colonic motility. It is suggested that in the rat sympathetic reflex inhibition of colonic motility caused by abdominal nociception is mediated via presynaptic alpha 2-adrenoceptors inhibiting the excitatory cholinergic neurons. PMID- 2990839 TI - [Ultrastructural study of primary hepatocellular carcinoma--hepatoma grading approach]. PMID- 2990840 TI - [Relation between breast cancer and myoepithelial cells]. PMID- 2990841 TI - [Primary malignant fibrous histiocytoma of the bone--report of a case and review of the literature]. PMID- 2990842 TI - [Differential method of analyzing ligand complex formation with binding centers. The detection of the superhigh-affinity binding centers of opiate ligands]. PMID- 2990843 TI - [Dynamics of changes in the spontaneous quantum and nonquantum secretion of acetylcholine from the motor nerve endings after nerve dissection]. PMID- 2990844 TI - [Side effects of antacids]. PMID- 2990845 TI - [Cytogenetics of juvenile tumors]. PMID- 2990846 TI - [Psoriasis in the light of basic research]. PMID- 2990847 TI - [Long-term cultivation of the Mazurenko leukemia virus in mouse and rat cell lines]. AB - Reproduction of the Mazurenko leukemia virus was studied in cell lines of different origin. Mongrel rat fibroblasts nonproductively transformed with variola vaccine virus proved to be the most adequate system for the virus replication. The leukemogenic virus reproduction in transformed rat cells was observed for 250 days. PMID- 2990848 TI - Heterogeneity of major affective disorders. Biological and clinical evidence. AB - In this paper we summarize the results of our recent and present research focused on analyzing the correlations between neurochemical, pharmacological and clinical parameters in patients with Major Depression. There is evidence that: a) pretreatment urinary MHPG is a useful predictor for clinical response to tricyclic antidepressants and to long-term lithium treatment; b) urinary MHPG is positively correlated to the age at onset of the disease; c) previous responses to tricyclics and age at onset of affective illness are supplementary tools for predicting the effectiveness of lithium and antidepressant drugs; d) platelet alpha-2-adrenoceptor density is inversely correlated with both urinary MHPG and age at onset; e) cerebral ventricular size is positively correlated with urinary MHPG and age at onset and may discriminate between patients with different outcomes on lithium prophylaxis; f) low MHPG excretors are more likely to have suffered from stressful life events in early childhood than normal-to-high excretors. Taken together, these results lend strong support to the hypothesis that Major Affective Disorder is a heterogeneous illness and that inherently different subgroups of affective patients can be recognized. PMID- 2990849 TI - Endocrine response to acute changes of brain blood flow due to lower body negative pressure in man. AB - The decrease of brain flow due to lower body negative pressure (LBNP) was used to study the role of cerebral glucopenia in the activation of anterior pituitary function in 10 young healthy men. During LBNP with a maximum negative pressure of -50 mm Hg the changes in heart rate, blood pressure cerebral blood flow (with the aid of rheoencephalography) and electrocardiogram were recorded and the levels of hGH, ACTH, hPRL, insulin and cortisol were measured with the aid of radioimmunoassay. During the first investigation an infusion of 20% glucose (1 g per min for first 30 min and 0.5 g per min for next 30 min) was used, while during the second investigation (one week later) the infusion of saline was applied. It was found that the infusion of glucose prevented the increase of hGH which was observed only during the infusion of saline. No differences in the level of ACTH, cortisol and hPRL were observed between the investigations with glucose and saline infusion. The increased level of insulin was related to the level of glucose during the infusion, while no changes were found during saline infusion. It was suggested that the increase of hGH level during LBNP resulted from glucopenia in cerebral tissue. PMID- 2990850 TI - Stimulation of the adenosine 3',5'-monophosphate and the Ca2+ messenger systems together reverse dopaminergic inhibition of prolactin release. AB - Dopaminergic inhibition of PRL release stimulated by agents that affect cytosolic Ca2+ concentrations, C-kinase activity, and cAMP levels was studied in perifused rat anterior pituitary cells cultured on cytodex beads. We used A23187 (20 microM) to increase intracellular Ca2+, the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA; 50 nM) to stimulate C-kinase, forskolin (10 microM) to increase intracellular cAMP, and 8-bromo-cAMP to mimic cAMP. Dopamine (10 microM) inhibited PRL release to 20-60% of the basal release within 10 min. After 30 min of preincubation with dopamine, the absolute amount of release stimulated by 100 nM TRH was strongly inhibited, although the pattern of release, a quick burst followed by sustained release at a lower rate, was the same in the presence or absence of dopamine. A23187 (20 microM) caused a rapid burst of PRL release that subsided within 10 min, and TPA (50 nM) caused a sustained release that began within 4 min and continued for at least 30 min. TPA and A23187 combined caused a rapid burst of release followed by a sustained phase of release similar to that caused by TRH. Preincubation with dopamine inhibited the absolute amount of PRL release caused by A23187 alone, TPA alone, or the two combined, although, as with TRH, the pattern of release remained the same. Forskolin (1 or 10 microM) or 8 bromo-cAMP (3 mM) induced a 1.5- to 2-fold increase in PRL release, and this was completely prevented by dopamine. Preincubation with both dopamine and 8-bromo cAMP or forskolin restored the amount of release stimulated by TPA alone or TPA and A23187 in the presence of dopamine to the level of release stimulated by these agents in the absence of dopamine. Therefore, activating either the cAMP messenger system or the Ca2+ system alone will not abolish dopaminergic inhibition, but activating the two together will. These results suggest that dopamine blocks release by inhibiting both adenylate cyclase and a step in the Ca2+ messenger system. PMID- 2990851 TI - Evidence that a benzodiazepine receptor mechanism regulates the secretion of pituitary beta-endorphin in rats. AB - The effects of agents representing three classes of benzodiazepine receptor acting drugs on circulating levels of beta-endorphin-like immunoreactivity (beta END-LI) were examined in male rats. The active benzodiazepine receptor antagonists, ethyl-beta-carboline-3-carboxylate (beta-CCE, 30 mg/kg), methyl-beta carboline-3-carboxylate (3 mg/kg), and 2-phenylpyrazolo [4,3-c]quinolin-3(5H)-one (CGS-8216, 3 mg/kg), all evoked 3- to 4-fold increases in plasma levels of beta END-LI as compared to control values. The beta-CCE-induced rise in circulating beta-END-LI was significantly attenuated by pretreatment with the agonist diazepam (2.5 mg/kg) and the antagonist ethyl-8-fluoro-5,6-dihydro-5-methyl-6-oxo 4H-imidazo-[1,5-alpha] [1,4]benzodiazepine-3-carboxylate (Ro 15-1788, 10 mg/kg) but was unaltered by morphine (1 and 5 mg/kg). Ro 15-1788 also significantly attenuated the methyl-beta-carboline-3-carboxylate- and CGS-8216-induced release of pituitary beta-END-LI in vivo. Morphine (5 mg/kg) and diazepam (5 mg/kg) together, but neither alone, significantly reduced the rise in plasma beta-END-LI due to physical immobilization or foot shock. Pretreatment with dexamethasone (100 micrograms), an inhibitor of pituitary anterior lobe (AL) beta-END-LI secretion, completely prevented the plasma beta-END-LI increase due to beta-CCE. Chromatographic analysis of plasma beta-END-LI revealed that most of the beta-END LI secreted in response to beta-CCE and CGS-8216 resembles beta-lipotropin (beta LPH), a marker for beta-END-LI release from the AL, in molecular size. Results of in vitro studies indicate that the effects of the anxiogenic agents, beta-CCE and CGS-8216, on AL beta-END-LI release in vivo were not mediated by direct actions of these agents on the pituitary gland. Together, these findings suggest that an interaction exists between a benzodiazepine receptor mechanism(s) and regulation of hypothalamic corticotropin-releasing factor(s) which in turn controls beta-END LI secretion from the AL of the rat pituitary gland. PMID- 2990852 TI - Effects of adenosine analogs on glucagon-stimulated adenosine 3',5'-monophosphate formation in Sertoli cell cultures from immature rats. AB - In the present study, we have examined the effects of two adenosine analogs, ( )N6-(R)phenyl-isopropyl-adenosine (PIA) and 2-chloro-adenosine, on glucagon- and FSH-stimulated cAMP production in Sertoli cell cultures isolated from immature (19-day-old) rats. Both FSH and glucagon caused a 5- to 10-fold stimulation of cAMP levels in the spent media from Sertoli cell cultures during an 18-h incubation. Addition of 1 microM PIA significantly inhibited both FSH- and glucagon-stimulated cAMP levels. In the presence of a maximal concentration of glucagon (2.5 micrograms/ml), PIA caused a concentration-dependent inhibition of cAMP formation, and the concentration of PIA causing half-maximal inhibition of cAMP formation (IC50) ranged from 0.5-1 nM. When Sertoli cells were incubated with increasing concentrations of glucagon (1.28 ng/ml to 4.00 micrograms/ml) in the absence and presence of either PIA (1.0 microM) or 2-chloro-adenosine (10.0 microM), the responses to glucagon, measured as cAMP formation, were almost completely abolished. 1-Methyl-3-isobutylxanthine (MIX), a well known inhibitor of cAMP phosphodiesterase activity, is also an inhibitor of adenosine binding to receptors on the cell membrane. When Sertoli cells stimulated with glucagon (2.5 micrograms/ml) were incubated in the absence and presence of MIX (0.1 mM) and increasing concentrations of PIA (0.025-10,000 nM), the presence of MIX reduced the inhibitory activity of PIA by almost 2 orders of magnitude (IC50 without MIX, 0.5 nM; IC50 with MIX, 20 nM). Thus, the present study shows that adenosine analogs inhibit agonist-stimulated cAMP formation in cultured Sertoli cells, and that MIX reduces this effect. This indicates that cultured Sertoli cells from immature rats contain A1-receptors for adenosine mediating inhibitory effects on adenylate cyclase. PMID- 2990853 TI - Superoxide radical generation, NADPH oxidase activity, and cytochrome P-450 content of rat liver microsomal fractions in an experimental hyperthyroid state: relation to lipid peroxidation. AB - The effect of thyroid hormone treatment on hepatic microsomal functions related to NADPH-dependent electron transfer reactions was studied in rats given 0.1 mg T3/kg BW for 1, 2, 3, and 7 consecutive days. This treatment resulted in increased rates of O2-. generation by microsomal fractions, concomitantly with an enhancement in NADPH oxidase activity and decreased cytochrome P-450 content, in livers exhibiting increased respiration. Subsequent studies showed elevated levels of malondialdehyde in microsomal fractions and liver homogenates, as well as augmented chemiluminescent response in the latter system. These results indicate that the calorigenic effect of T3 on the liver tissue is accompanied by a stimulation of microsomal functions involving univalent reduction of oxygen. This cellular response might lead to a greater lipid peroxidative rate and cytochrome P-450 loss as secondary events of thyroid hormone action. PMID- 2990854 TI - Purification of thyroid lysosomes by colloidal silica density gradient centrifugation. AB - A procedure was devised for fractionating crude thyroid lysosomal particles (P750 15,000) by self-forming density gradient centrifugation with colloidal silica. Two discrete particle-containing peaks were observed, based on 131I-labeling and acid phosphatase activity: a heavy peak (density, 1.11-1.12) and a light peak (density, 1.05). Ultrastructural analysis revealed that the heavy peak consisted almost entirely of lysosomes, whereas the light peak represented a heterogeneous mixture of small vesicles and fragments of other intracellular organelles. In thyroids removed from rats 30 min after 131I injection, almost all of the 131I was present in the low density peak. This 131I appeared on sucrose density gradient centrifugation as a 19S peak, and it was almost completely insoluble in trichloroacetic acid. This was interpreted as indicating that the low density peak contained pinocytotic vesicles. In thyroids removed 4 days after 131I injection, the radioactivity appeared largely in the high density peak. Both the trichloroacetic acid solubility and the pattern on sucrose density gradient centrifugation indicated that the [131I] thyroglobulin had undergone extensive proteolysis. Thyroglobulin proteolytic activity was found primarily in the high density particles and to only a small extent in the low density particles. Studies performed at intervals after 131I injection combined with double labeling (131I and 125I) experiments provided evidence that radioactivity was transferred from the low density to the high density particles. Heterogeneity existed within the dense peak, related to the degree of thyroglobulin degradation, as it was observed that thyroid lysosomes become denser with increasing proteolysis of thyroglobulin. The acid phosphatase in the low density particles could be distinguished from that in the high density (lysosomal) particles by its elution pattern on Sephadex G-200 column chromatography, its response to freezing and thawing, and its reactivity with p-nitrophenylphosphate. It was concluded, therefore, that the acid phosphatase in the low density fraction was derived from prolysosomal structures such as vesiculated Golgi-endoplasmic reticulum lysosomes. The prolysosomal acid phosphatase associated with the low density fraction appeared to be a large membrane-bound molecule which could be transformed into lysosomal acid phosphatase by incubation at pH 5.0. PMID- 2990856 TI - Polypeptide hormone receptor phosphorylation: is there a role in receptor mediated endocytosis of human growth hormone? AB - To determine whether receptor phosphorylation is a critical step in the internalization of polypeptide hormones and their receptors, we have studied a model system wherein insulin stimulates phosphorylation of its receptor and is also internalized. Using insulin as a positive control, we found that it stimulated a partially purified plasma membrane preparation of IM-9 lymphocytes to autophosphorylate its receptor and to catalyze the phosphorylation of a tyrosine-containing substrate. The human GH (hGH) receptor of the IM-9 lymphocytes, when coupled to [125I]iodo-hGH, migrated as a 140,000-dalton protein on polyacrylamide gel electrophoresis. This protein, in contrast to the insulin receptor, was not phosphorylated by the addition of hGH, nor did hGH stimulate this preparation to phosphorylate the tyrosine-containing substrate poly (GluNa,Tyr)4:1, casein, or histone f2b under a variety of conditions. We conclude that receptor phosphorylation is not a critical intermediate in the receptor mediated endocytosis of hGH and probably other polypeptide hormones and growth factors. PMID- 2990855 TI - Alpha 1-adrenoreceptors and alpha 1-adrenoreceptor-mediated thyrotropin release in cultures of euthyroid and hypothyroid rat anterior pituitary cells. AB - TSH responses to adrenergic agonists have been measured in 3-day monolayer cultures of euthyroid and hypothyroid male rat anterior pituitary (AP) cells. Responses were qualitatively similar in that (-)epinephrine and (-)norepinephrine had the same ED50 in each culture (ED50 = approximately 6 and 16 nM, respectively) and demonstrated the same alpha 1-adrenergic specificity. Hypothyroid cultures secreted approximately twice as much TSH per cell as euthyroid cultures over the 2-h experimental period. (-)Epinephrine produced a 95 +/- 8% (mean +/- SE) release of TSH relative to basal secretion in euthyroid cultures and only 62 +/- 7% release in the hypothyroid cultures (P less than 0.01). The comparable figures for (-)norepinephrine were 62 +/- 7% and 38 +/- 5%, respectively (P less than 0.05). In absolute terms, adrenergic agonists released the same amount of TSH from euthyroid and hypothyroid cultures. In contrast, TRH (and the Ca+2 channel ionophore A23187) released twice as much TSH from the hypothyroid cells as in the euthyroid cultures. Epinephrine-induced TSH release was significantly impaired (P less than 0.001) when either euthyroid or hypothyroid cells were cultured without thyroid hormones. In contrast, TRH induced TSH release was enhanced (P less than 0.001) in the euthyroid cultures. [3H]Dihydroergocryptine [( 3H]DHE) was used to quantify alpha 1-adrenoreceptors on the same cell preparations as those used to derive the functional data (see above). Prazosin (1 microM) was used to define nonspecific binding of [3H]DHE. Specific binding to euthyroid cells had a Kd of 5.8 +/- 4 nM and a maximum binding capacity of 2.2 +/- 0.4 fmol/10(5) cells (n = 5). In parallel cultures of hypothyroid cells, the Kd (6.2 +/- 5 nM) was not significantly different, whereas the maximum binding capacity (1.4 +/- 0.3 fmol/10(5) cells) was significantly reduced (P less than 0.05). Adrenergic compounds showed a rank order of potency of prazosin greater than (-)epinephrine greater than or equal to ( )norepinephrine greater than or equal to yohimbine greater than clonidine against the binding of 5 nM [3H]DHE to euthyroid and hypothyroid cells. The amount of [3H]DHE binding per cell that each adrenergic compound was able to displace at saturating concentrations was less in hypothyroid cells than in euthyroid cells. There was no change in the ED50 values of these compounds in the same experiments.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990857 TI - Measurement of growth hormone and prolactin receptor turnover in rat liver. AB - To study the rate of disappearance of GH and PRL receptors in the livers of rats treated with cycloheximide, a technique has been devised for multiple sampling from the liver of each anesthetized rat. In rats treated with cycloheximide (1 or 5 mg/kg, iv), binding sites for both bovine GH and ovine PRL disappeared following first order kinetics over the 2-h sampling period. The half-time for the GH receptor was 30-40 min, equivalent to a rate constant of approximately 0.02 min-1. The half-time for the PRL receptor was 40-50 min, equivalent to a rate constant of approximately 0.015 min-1. At 0.1 mg/kg cycloheximide, slower disappearance rates were seen for both receptors, and the GH receptor showed a partial recovery. Over the same period, binding sites for insulin were unaltered at any cycloheximide dose. Assuming cycloheximide acts simply to inhibit new receptor synthesis, these rates represent the turnover time for GH and PRL receptors in rat liver. PMID- 2990858 TI - Dexamethasone permits the release of an inhibitor of pyruvate dehydrogenase from Reuber H-35 hepatoma cells in response to insulin. AB - Reuber H-35 rat hepatoma cells respond to physiological levels of insulin as a growth factor. Glucocorticoids antagonize this response. A chemical mediator of insulin action which activates mitochondrial pyruvate dehydrogenase has also been isolated from these cells. The present report demonstrates that if the H35 cells are incubated with glucocorticoid before treatment with insulin, they produce not only the stimulator, but also inhibitory mediator. Cells exposed to the glucocorticoid but not to insulin do not produce the inhibitory mediator. Therefore, insulin interaction with the cell is necessary to elicit this negative modifier of pyruvate dehydrogenase. A time course of the response suggests that the effect of the glucocorticoid is time dependent. The inhibitory mediator can be separated from the stimulatory mediator by molecular sieve chromatography. These results suggest a biochemical basis for glucocorticoid-mediated insulin resistance. PMID- 2990859 TI - Photoaffinity labeling of pituitary and gonadal receptors for gonadotropin releasing hormone. AB - A radioactive photoaffinity label for the GnRH receptor was prepared by derivatization of radiodinated [D-Lys6] des-Gly10-GnRH N-ethylamide with the heterobifunctional photolabile reagent N-hydroxysuccinimidyl-4-azido-benzoate. This high affinity photoreactive analog was employed for radiolabeling and characterization of pituitary GnRH receptors in rat, rabbit, mouse, sheep, and cow adenohypophyses and gonadal GnRH receptors in the rat ovary and testis. In rat, rabbit, and mouse pituitary glands, analysis of the GnRH receptor-ligand complex by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography showed two labeled components, both of which were displaced by unlabeled GnRH agonist and antagonist analogs. The larger receptor component was a relatively broad band, with mol wt in rat, rabbit, and mouse of 59,000 +/- 1,900, 62,000 +/- 700, and 60,000 +/- 800, respectively. In the rat pituitary gland, the larger component was composed of 63,000 and 52,000 mol wt components, of which the latter was more heavily labeled and was predominant in purified pituitary gonadotrophs. The mol wts of the smaller components were 40,000 +/- 1,600, 43,000 +/- 1,200, and 41,000 +/- 1,000, respectively. In bovine and ovine pituitary glands, the photolabeled GnRH receptor was a single band with mol wt of 42,000 +/- 1,200 and 39,000 +/- 500, respectively. In the rat ovary and testis, photolabeled GnRH receptors were similar to those in the rat pituitary gland, with two distinct components of 53,000 +/- 1,000 and 42,000 +/- 1,000 mol wt. These findings demonstrate that the pituitary receptors that mediate similar physiological actions of GnRH in different species possess broadly similar structural properties, with minor variations between species. It is also evident that the divergent actions of GnRH in different tissues of the same species, as in the rat pituitary and gonads, are expressed through receptors of similar structure. PMID- 2990860 TI - Evidence for two populations of masked gonadotropin-binding sites in the corpus luteum of the rhesus monkey (Macaca mulatta). AB - To evaluate the possible existence of masked gonadotropin receptors in the corpus luteum, we characterized the effects of alcohols and neuraminidase on [125I]iodohuman LH binding to in vitro preparations of luteal tissue from the rhesus monkey and pseudopregnant rat. The presence of 1-8% (vol/vol) ethanol enhanced specific LH binding to macaque luteal particulates under steady state conditions (25 C, 20-h incubation), with a maximal effect at 8% ethanol (166% of control uptake; P less than 0.05). However, 1-8% ethanol had no effect on LH binding to rat luteal tissue. Higher concentrations of ethanol (20%) decreased LH binding relative to control values in both species. Ethanol modulation of LH binding to macaque luteal particulates and dispersed cells was a time- and temperature-dependent process. At 4 and 25 C, ethanol increased LH uptake at all times during a 32-h incubation. However, at 37 C, ethanol increased LH uptake at 30 min; binding peaked at 2 h and then returned to control levels within 20 h. The optimal concentration of ethanol for enhancing LH uptake was inversely related to the incubation temperature. The increase in LH binding to macaque luteal particulates in the presence of ethanol was reversible; binding returned to control levels if ethanol was removed before the addition of labeled LH. Longer straightchain alcohols (butanol, pentanol, and octanol) were progressively more potent than ethanol in enhancing LH binding to macaque luteal particulates and dispersed luteal cells. Pretreatment of luteal particulates from either the rat or monkey with neuraminidase increased LH uptake, with a maximal effect (160% of control) at 1 mg/ml enzyme. Scatchard analyses revealed that both ethanol and neuraminidase increased (P less than 0.05) the number of LH-binding sites without altering the affinity for gonadotropin. Moreover, the effects of ethanol and neuraminidase were additive, i.e. increased LH binding during combination of the two treatments approximated the sum of the individual effects. The data suggest that two distinct populations of LH-binding sites are masked within the membranes of the monkey corpus luteum. The ability of two markedly different agents, alcohol and neuraminidase, to increase LH binding indicates that diverse mechanisms may modulate the masking/unmasking of gonadotropin receptors in target cell membranes. Finally, the inability of ethanol to enhance LH binding in the rat suggests species differences in the receptor population or milieu of luteal membranes. PMID- 2990861 TI - Modulation of membrane fluidity in the primate (Macaca mulatta) corpus luteum: correlation with changes in gonadotropin binding. AB - Addition of alcohols to particulate or cellular preparations of the monkey corpus luteum unmasks gonadotropin-binding sites via a temperature-sensitive process. Since alcohols and temperature are known modulators of membrane fluidity, we measured the fluidity of luteal membranes and determined whether the effects of ethanol and temperature on gonadotropin binding correlated with changes in the fluid state of the membrane. The fluidity of membranes from the macaque and rat corpus luteum was estimated from the fluorescence polarization of the lipophilic membrane probe 1,6-diphenyl-1,3,5-hexatriene (DPH). The absorption and emission spectra of DPH incorporated into luteal membranes were typical of those in other systems. Fluorescence intensity increased rapidly during the first 60 min of incubation and reached steady state conditions within 3 h. In contrast, polarization was constant within minutes and was insensitive to pH, ionic strength, tissue concentration, or DPH levels over the ranges tested. Fluorescence polarization was acutely sensitive to the temperature of the assay medium; polarization decreased as temperature increased from 4-50 C, and no phase transitions were observed. Addition of 4-20% and 8-20% ethanol to monkey and rat membranes, respectively, decreased (P less than 0.05) polarization relative to control values. However, ethanol was less effective on rat membranes, such that 20% ethanol was required to elicit a similar change in polarization as 8% ethanol in macaque membranes. The decrease in polarization was reversed to control levels when ethanol was removed from the incubation medium. Changes in fluorescence polarization of DPH-labeled macaque membranes elicited by ethanol and temperature correlated significantly (r = -0.97) with changes in specific [125I]iodohuman LH binding. In contrast, pretreatment of luteal membranes from the monkey and rat with neuraminidase, which unmasks another population of LH-binding sites in both species, did not alter polarization. We conclude that the fluorescence polarization of DPH is a useful tool for estimating membrane fluidity in the corpus luteum. Furthermore, changes in membrane fluidity may play an important role in the masking/unmasking of alcohol-sensitive (but not neuraminidase sensitive) gonadotropin-binding sites in the macaque corpus luteum. Finally, the lesser effects of ethanol in the rat suggest important species differences in the receptor milieu and composition of luteal membranes. PMID- 2990862 TI - Posttranslational processing of proadrenocorticotropin/endorphin-derived peptides during postnatal development in the rat pituitary. AB - The anterior pituitary content of pro-ACTH/endorphin-related peptides increased 5 fold from birth to 4 weeks and increased another 3-fold by adulthood. In contrast, the neurointermediate lobe content of pro-ACTH/endorphin-related peptides increased 15-fold from birth to 4 weeks and another 10-fold by adulthood. Despite the dramatic increase in content, posttranslational processing of pro-ACTH/endorphin in the neurointermediate lobe of the neonate closely resembled intermediate lobe processing in the adult; alpha MSH- and beta endorphin-sized molecules (rather than ACTH and beta-lipotropin) accounted for more than 90% of the immunoreactivity in both neonates and adults. In the neurointermediate pituitary of both the neonate and the adult, the alpha MSH sized material was largely diacetylated, and the beta-endorphin was both alpha-N acetylated and C-terminally shortened. However, the extent of C-terminal shortening of beta-endorphin in the neurointermediate lobe of the neonate was not as great as that observed by postnatal day 21 or that in the adult. In the anterior pituitary, distinct differences in processing occurred between birth and adulthood. Proteolytic processing of pro-ACTH/endorphin was not as extensive on day 1 as in the adult, and pro-ACTH/endorphin accounted for 40-50% of the total immunoreactive peptide. The extent of processing of precursor increased around day 21, and a higher percentage of ACTH-(1-39) and beta-endorphin-(1-31)-sized material was found. Neonatal anterior pituitary contained substantial amounts of alpha MSH-sized material, whereas in adult anterior pituitary, less than 1-2% of the ACTH-related material was alpha MSH-sized. Despite these differences in the extent of proteolytic processing, neonatal anterior pituitary corticotropes resembled those of adults, in that they did not alpha-N-acetylate beta-endorphin or alpha MSH. Immunocytochemical studies demonstrated that a subset of the neonatal anterior pituitary corticotropes produced alpha MSH-related molecules. PMID- 2990863 TI - A simple and rapid method using polyethylenimine-treated filters for assay of solubilized LH/hCG receptors. AB - Filtration through polyethylenimine (PEI)-treated membrane filters efficiently separates free radiolabeled human (h) LH or hCG from solubilized receptor-hormone complexes. Solubilized LH/hCG receptors of bovine, porcine, and rat origin are suitable for measurement by this method. Specific binding assayed by the PEI filter technique was 106-138% of specific binding assayed by double polyethylene glycol precipitation, demonstrating that the method gave quantitative recovery of bound hormone. Nonspecific binding was consistently lower with PEI-treated filters. Intra- and interassay variability for the two different methods was similar. These results indicate that filtration through PEI-treated membrane filters is a simple and rapid method for assay of solubilized LH/hCG receptors. PMID- 2990864 TI - Sympathetic adrenal denervation decreases adrenal blood flow without altering the cortisol response to hemorrhage. AB - To test whether or not adrenal sympathetic innervation is required for the adrenocortical response to small hemorrhage, awake dogs were studied after unilateral adrenal sympathetic denervation. Bilateral adrenal vein cannulas were placed chronically to permit measurement of cortisol, epinephrine, and norepinephrine secretion rates and adrenal blood flow simultaneously from the intact and the denervated adrenal. Plasma ACTH concentration was measured and the presentation rate of ACTH was calculated as the product of plasma ACTH concentration and adrenal plasma flow. Unilateral isolation of the sympathetic chain from the spinal cord at thoracic levels 9-12 (T9-12) had no effect on adrenal blood flow, on the presentation rate of ACTH, or on cortisol secretion after 10 mg/kg hemorrhage. However, thoracic levels 9-12 denervation prevented the secretory response of catecholamines to hemorrhage without lowering basal catecholamine secretion. Unilateral splanchnicotomy, the sectioning of the thoracic and upper lumbar splanchnic nerves, reduced adrenal blood flow and the presentation rate of ACTH, suppressed basal catecholamine secretion, and prevented the catecholamine response to hemorrhage. However, there was no reduction in the secretory response of cortisol to 10% or 20% hemorrhage. These findings suggest that in the absence of sympathetic innervation to the adrenal, increases in adrenal sensitivity to ACTH occur to offset decreased ACTH presentation rate resulting in a normal cortisol response to hemorrhage. However, adrenal sensitivity to exogenous ACTH was not increased in non-hemorrhaged dogs after unilateral splanchnicotomy. Thus, hemorrhage must activate a non-ACTH mechanism that is independent of sympathetic adrenal innervation to augment adrenal sensitivity to ACTH. Sympathetic innervation to the adrenal has profound effects on catecholamine secretion and on adrenal blood flow but is not required for the secretory response of cortisol to small hemorrhage. PMID- 2990865 TI - Identification of subpopulations of rat granulosa cells: sedimentation properties and hormonal responsiveness. AB - Ovarian granulosa cells from small follicles have generally been considered to comprise a homogeneous cell population; however, stratified arrangements of hormone receptors have been found in antral and mural granulosa cells of Graafian follicles. Using cultured granulosa cells derived from immature, hypophysectomized, estrogen-treated rats, we have previously shown that vasoactive intestinal peptide (VIP) as well as FSH stimulates steroid production by these cells. Dose-response analysis indicated that the actions of these hormones were additive, suggesting the presence of subpopulations of granulosa cells. Using a continuous (0-30%) Metrizamide density gradient, we have identified three populations of granulosa cells with different sedimentation properties. After centrifugation for 15 min at 1500 X g, cells sedimented at Metrizamide concentrations of 13%, 18%, and 20% (peaks A, B, and C, respectively). The subpopulation with lowest density (peak A) comprised 5% of the total cells, whereas the remaining cells were distributed approximately equally in the other two peaks. The profiles of estrogen and progesterone production by these cells in response to FSH and VIP indicated that FSH preferentially stimulated steroid production in cells with the highest density (peak C), whereas VIP mainly induced steroidogenic responses in cells of intermediate density (peak B). In contrast, cells with the lowest density (peak A) were unresponsive to either hormone. Treatment with forskolin, a universal adenylate cyclase activator, induced steroid production in both subpopulations B and C. Further studies demonstrated that LH/human CG receptors were induced by FSH and forskolin in cells from peak C, whereas VIP treatment did not induce LH/human CG receptors in cells from peak B. In unfractionated cultured cells, GnRH potently antagonized FSH- but not VIP-induced steroidogenesis. Upon density-gradient fractionation, the profile of GnRH receptor content correlated well with GnRH effects since FSH responsive cells (peak C) contained the majority of GnRH receptors. The present results demonstrate that granulosa cells from immature follicles are heterogeneous and consist of two major subpopulations of cells with differential responsiveness to FSH and VIP. These findings provide the basis for further morphological and biochemical analysis of subpopulations of granulosa cells during follicular development. PMID- 2990866 TI - Rat pineal N-acetyltransferase activity: stimulation, exhaustion, and recovery. AB - Rat pineal N-acetyltransferase activity (NAT) has a circadian rhythm, with a peak in the dark time. The rhythm of NAT was studied in young rats (26-40 days old). In particular, the possibility of maintenance of prolonged low NAT or prolonged high NAT was studied. NAT (given in nanomoles per pineal gland/h) had peak activity (33.0) in the dark time. Forty-eight hours of constant light suppressed the peak and maintained low NAT (0.5 or less). In vivo, in constant light, NAT was stimulated (to 13.1-29.4) by single injections of isoproterenol. Similarly, NAT was stimulated by isoproterenol or (Bu)2cAMP in vitro. In vivo, multiple injections of isoproterenol stimulated NAT for 14 h (to peak 31.9), but NAT was exhausted and fell to less than 5 over the next 8 h. Similarly, isoproterenol and (Bu)2cAMP exhausted NAT in vitro. High NAT was not maintained. NAT recovered [could be restimulated by dark in vivo, isoproterenol in vivo or in vitro, or (Bu)2cAMP in vitro]. Exhaustion was not permanent. Light can keep NAT at low levels, but dark cannot maintain peak NAT beyond 14 h. The results are interpreted as meaning that the duration of NAT is fixed. PMID- 2990867 TI - Mainly mu-opiate receptors are involved in luteinizing hormone and prolactin secretion. AB - We evaluated plasma PRL and LH concentrations in the rat after the administration of drugs that exert a specificity directed mainly, although not absolutely, toward the mu-, delta-, or kappa-opiate receptors, in order to investigate the role of different receptors and thus the respective endogenous ligands in the modulation of the release of these anterior pituitary hormones. LH concentrations were evaluated in prepuberal female rats, in adult male rats, and in ovariectomized, estradiol benzoate-treated rats. PRL concentrations were evaluated in suckling rats, in ovariectomized, estradiol benzoate-treated rats, and in ether-stressed rats. The delta-antagonist ICI 154129 never affected PRL or LH concentrations, whereas both the mu- and kappa-antagonists, naloxone and MR 1452, respectively, seemed to be effective. However, when graded doses of the two classes of antagonists were tested, the mu-antagonist appeared to be effective on both hormones at doses that were one tenth of those of the kappa-antagonist. In conclusion, the mu-receptor seems to be the most profoundly involved in the regulation of PRL and LH secretion. PMID- 2990868 TI - Aromatase inhibitors prevent granulosa cell differentiation: an obligatory role for estrogens in luteinizing hormone receptor expression. AB - To determine the role of newly synthesized estrogens in LH receptor expression, granulosa cells from diethylstilbestrol-implanted immature rats were cultured with FSH plus aromatase inhibitors. When present throughout the 48-h culture period, 4-hydroxy-4-androstene-3,17-dione (4-OHA; greater than or equal to 100 microM) and 1,4,6-androstatriene-3,17-dione (greater than or equal to 5 microM) inhibited FSH-induced LH receptor formation by 40% and 90%, respectively. Both aromatase inhibitors caused relatively greater inhibition of LH receptor formation when added from 20-48 h of culture, the period during which FSH stimulated estrogen synthesis occurs (85% maximal inhibition with 4-OHA and 95% with 1,4,6-androstatriene-3,17-dione). Addition of estradiol, but not androstenedione, reversed the reduction of LH receptor formation by 4-OHA, indicating that the effects of the aromatase inhibitors were specifically related to their blockade of estradiol synthesis. The stimulation of estrogen production by FSH alone (8-fold) or with androstenedione (80-fold) during the 48-h culture period was prevented by 4-OHA. FSH-stimulated cAMP production was initially enhanced by 4-OHA from 0-20 h of culture, but was reduced from 20-48 h. Lower concentrations of 4-OHA (less than or equal to 50 microM) amplified FSH stimulated cAMP production and LH receptor formation. However, these responses were blocked by the antiestrogen keoxifene or the antiandrogen flutamide, indicating that 4-OHA or a metabolite may have partial estrogenic or androgenic properties. The inhibitory effects of higher concentrations of 4-OHA on LH receptor expression were potentiated by keoxifene or flutamide. These results indicate that estrogen production and action are necessary for LH receptor expression in the granulosa cell. PMID- 2990869 TI - Characterization of the receptors for insulin and the insulin-like growth factors on micro- and macrovascular tissues. AB - Insulin and insulin-like growth factors (IGFs) have been implicated in the pathogenesis of diabetic retinopathy and peripheral vascular complications. Previously, we have shown that retinal capillary endothelial cells responded to insulin and IGFs for metabolic and growth effects, whereas aortic endothelial cells were not responsive. In contrast, vascular supporting cells from both retinal capillaries (i.e. pericytes) and aorta (i.e. smooth muscle cells) responded equally to insulin, IGF-I, and IGF-II. The structure and ligand specificities of the receptor for these peptides were studied by covalently cross linking 125I-labeled peptide hormones to their respective receptors using disuccinimidyl suberate, followed by polyacrylamide gel electrophoresis and autoradiography. The binding subunit of the insulin receptor, alpha-subunit, for all cell types was found to have a mol wt 145,000 under reduced conditions. Labeling of this band was inhibited by 10(-9) M insulin, antiinsulin receptor antibodies, and 10(-8) M IGF-I, but not by multiplication-stimulating activity (IGF-II). The beta-subunit of the insulin receptor in endothelial cells was identified by its ability to be autophosphorylated when stimulated by insulin and was found to have a mol wt of 99,000. Covalent cross-linking of IGF-I to its receptor revealed a mol wt of 145,000, similar to that of insulin receptor, except that IGF-I was 100-fold more potent than insulin in competing with [125I]IGF-I for binding. [125I]IGF-II in all cells was cross-linked to receptor with mol wt of 260,000 and 230,000 under reduced and nonreduced conditions, respectively. IGF-I competed weakly with [125I]IGF-II, whereas insulin was ineffective. [125I]IGF-II also bound to the band with alpha mol wt of 135,000, which was inhibited by insulin, IGF-I, and IGF-II. In summary, receptors for insulin, IGF-I, and IGF-II on cells from micro- and macrovessels are biochemically similar to those in other cells. Interestingly, the finding of large numbers of IGF-I and IGF-II receptors on endothelial cells suggests that these growth factors play a physiological role and are involved in vascular complications associated with diabetes. PMID- 2990870 TI - Rat parathyroid hormone-(1-34) fragment: renal adenylate cyclase activity and receptor binding properties in vitro. AB - The rat PTH molecule contains five sequence differences from either the bovine or the human hormone within the biologically active 1-34 region. A synthetic rat 1 34 peptide was tested for activity by in vitro activation of canine and rat renal adenylate cyclase and binding to canine renal membrane receptors. The mean potency of 21,400 Medical Research Council units/mg in the canine adenylate cyclase system and 24,900 in the rat system was 8- to 10-fold higher than human 1 34 and 2- to 4-fold greater than bovine 1-34. These values represent the highest potency we have observed to date for a PTH preparation by these assay systems. In contrast, receptor binding of the rat fragment was comparable to that of bovine and human 1-34. Half-maximal inhibition of radioligand binding occurred at 1.7- 2.0 X 10(-9) M with all synthetic hormones. Hence, the amino acid substitutions in rat 1-34 appear to affect the cyclase-activating sequence domain without increasing avidity for the receptor. Analogs combining the rat sequence with modifications known to enhance receptor binding and/or retard enzymatic degradation offer a promising approach to the preparation of still more potent parathyroid agonists. PMID- 2990871 TI - The role of ascorbic acid in the function of the adrenal cortex: studies in adrenocortical cells in culture. AB - To investigate the role of ascorbic acid in the function of the adrenal cortex, we studied the effects of ascorbate on the regulation of 11 beta-hydroxylase in culture. When primary bovine adrenocortical cells were cultured in a serum-free defined medium in the absence of ACTH, 11 beta-hydroxylase activity declined with a half-time of about 40 h. When 50 microM cortisol, which acts as a pseudosubstrate for 11 beta-hydroxylase, was added to such cultures, 11 beta hydroxylase activity declined with a half-time of about 6 h. Ascorbate (5 mM) markedly reduced the rate of loss of 11 beta-hydroxylase activity in the presence of cortisol. Previous studies showed that phenolic and sulfoxide antioxidants, which also prevent loss of 11 beta-hydroxylase activity, inhibited the enzyme at concentrations somewhat higher than those required for protective activity. However, ascorbate at concentrations from 10 microM to 5 mM did not inhibit 11 beta-hydroxylase. The same range of ascorbate concentrations added to cells during a 24-h preincubation with cortisol showed increasing prevention of loss of 11 beta-hydroxylase activity. Ascorbate and a lowered concentration of oxygen were synergistic in their protective action. At 2% oxygen, 5 mM ascorbate almost completely prevented loss of 11 beta-hydroxylase activity in the presence of 50 microM cortisol. 11 beta-Hydroxylase activity was reinduced over a period of 5 days in third passage cultures by addition of 1 microM ACTH in defined lipoprotein-free medium. Addition of ascorbate enhanced the reinduction about 2 fold. The action of ascorbate in prevention of pseudosubstrate-mediated loss of activity and in enhancing reinduction of 11 beta-hydroxylase is specific; neither alpha-tocopherol nor selenium prevented loss of 11 beta-hydroxylase in the presence of cortisol or enhanced reinduction of 11 beta-hydroxylase in the presence of ACTH. As an additional test of specificity, it was shown that reinduction of 17-hydroxylase activity was completely unaffected by ascorbate, selenium, or alpha-tocopherol, and addition of cortisol to cultures with high 17 hydroxylase did not result in any loss of enzyme activity. Thus, a major function of ascorbate in the adrenal cortex is as a protective compound for cytochrome. PMID- 2990873 TI - Pancreatic hormone receptors on islet cells. AB - To assess whether islet cells are equipped with recognition units which allow an intra-islet regulation via released hormones, the presence of insulin and glucagon receptors is investigated on purified pancreatic A and B cells. Mono [125I]glucagon is shown to bind specifically to islet B cells, with similar binding characteristics as in isolated hepatocytes but involving less receptors per cell (2.10(4) per B cell vs. 8.10(5) per liver cell). Binding is half maximally displaced by 5.10(-9) M glucagon, a concentration known to induce half maximal biological effects in isolated B cells. These results are compatible with a regulatory role of glucagon in the insulin release process. No specific binding of [125I]tyr-A14-insulin is detected on pancreatic A cells. In order to increase receptor assay sensitivity, [123I]tyr-A14-insulin is prepared with at least 5 fold higher specific activity. Its validity for in vitro receptor analysis is demonstrated in IM-9 lymphocytes, where insulin binding is detectable down to 10(4) cells/ml. However, no insulin-binding sites are identified on pancreatic A cells, even at 10(6) cells/ml. If isolated A cells contain high affinity insulin receptors, their number should be inferior to 400 per cell, which is 50- to 500 fold lower than in classical insulin target cells. These findings explain the insensitivity of the glucagon release process to acute exposure to insulin. PMID- 2990872 TI - Synthetic rat atrial natriuretic factor inhibits in vitro and in vivo renin secretion in rats. AB - We investigated the action of a synthetic rat atrial natriuretic factor (ANF) with 28 amino acids on renin secretion in rats. Renin release by kidney cortex slices was determined after 90 min of incubation at 37C. ANF inhibited basal renin release in a dose-related fashion. ANF also decreased cAMP release and increased cGMP release in a dose-dependent manner. Renin release stimulated by 10(-7) M isoproterenol was inhibited by ANF with an ID50 of 5.8 x 10(-8) M. The renin-inhibitory effect was not calcium-dependent. In anesthetized rats, a bolus IV dose of ANF decreased plasma renin activity and cAMP concentration, but increased cGMP concentration. These data suggest that ANF inhibits renin secretion via the direct action on juxtaglomerular cells and that this effect may be partly mediated by the changes in cyclic nucleotide production. PMID- 2990874 TI - Polyphloretin phosphate, an antagonist of thyrotropin action: evidence for an interaction with the hormone rather than the receptor. AB - Polyphloretin phosphate (PPP) is known to be an inhibitor of bovine TSH (bTSH) induced stimulation of the thyroid in both in vivo and in vitro assays. The present studies were undertaken to delineate the mechanism of these effects. A high molecular weight PPP preparation strongly inhibited both the binding of 125I labeled bTSH [( 125I]bTSH) to human thyroid membranes and the stimulation of adenylate cyclase evoked by bTSH therein. Inhibition of bTSH-induced adenylate cyclase activity by PPP was evident both in the absence and the presence of NaCl (150 mM) in the incubation medium. Incubation of membranes with PPP, followed by its removal, did not affect subsequent binding of [125I]bTSH, indicating that PPP did not bind firmly to or damage the TSH receptor. Gel chromatography on Sephadex G-100 revealed that [125I]bTSH incubated with PPP eluted earlier than [125I]bTSH alone, indicating that PPP had formed a higher molecular weight complex with [125I]bTSH. This effect could be prevented by the addition of an excess of unlabeled bTSH to the incubation mixture. Binding of [125I] bTSH in the higher molecular weight peak generated by incubation with PPP was less than half that in control specimens of [125I]bTSH. Studies with PPP were also conducted in a highly sensitive assay that employs cultured porcine thyroid cells and measures the cAMP response induced by bTSH. The inhibitory effect of PPP on bTSH-induced cAMP accumulation was also evident in this assay. However, the presence of divalent cations Ca++ and Mg++ in the assay medium greatly diminished the inhibitory effect of PPP. Similarly, addition of Ca++ and Mg++ to the incubation medium greatly reduced or abolished the inhibitory effect of PPP on [125I]bTSH binding. Both effects of these salts to lessen the inhibitory response to PPP were overcome by increasing the PPP concentration. Gel chromatographic studies revealed that Ca++ and Mg++ acted by inhibiting the formation of the high molecular weight complex of bTSH and PPP. From these findings, we conclude that PPP exerts its inhibitory effect on TSH-induced stimulatory responses in the thyroid, in vivo as well as in vitro, by forming a complex with the hormone. The complex either does not bind to TSH receptors or does so with much lower affinity. PMID- 2990875 TI - Fibronectin as a marker of granulosa cell cytodifferentiation. AB - The hormonal regulation of fibronectin secretion by rat granulosa cells in culture was investigated: fibronectin was measured by a competitive enzyme-linked immunoadsorbant assay. Granulosa cells isolated from 25-day-old diethylstilbestrol-primed rats and cultured under defined conditions in the absence of hormones secreted low levels of fibronectin during the first 24 h of culture, after which there was a rapid increase in secretion until 72 h. In contrast, cultures treated with a combination of NIH-FSH-15 (200 ng/ml) and insulin (5 micrograms/ml) secreted low levels of fibronectin throughout the culture period. Subsequently, it was found that both FSH and insulin could independently suppress the increase in fibronectin secretion found in control cultures. Combined treatment with FSH and insulin resulted in a level of fibronectin which was the same as either FSH or insulin alone. The actions of FSH and insulin were dose dependent; 10 ng FSH/ml and 2.5 micrograms insulin/ml were required to produce a maximum suppression. The ability of (Bu)2cAMP (1.0 mM) to suppress fibronectin secretion suggested that the action of FSH on this parameter was mediated via the production of cAMP. Testosterone and estrogen alone did not influence secretion and did not modulate the actions of FSH and insulin. At the time at which FSH induces the cytodifferentiation of granulosa cells in culture, assessed by the increase in aromatase activity, fibronectin secretion is suppressed. The inverse relationship between fibronectin secretion and the induction of those granulosa cell functions essential for the development of the preovulatory follicle indicates that fibronectin may provide a useful marker for the stage of cytodifferentiation and follicular maturation. PMID- 2990876 TI - The effect of various corticotropin-releasing factor trains on the release of adrenocorticotropin, beta-endorphin, and beta-lipotropin from perifused ovine pituitary cells. AB - The dynamics of the release of proopiomelanocortin-derived hormones from ovine anterior pituitary cells in response to varying pulse characteristics of ovine corticotropin-releasing factor (CRF) were investigated with an in vitro automated 15-column simultaneous perifusion system. Columns of cells were stimulated continuously or with trains of CRF pulses of varying pulse length (2-16 min), pulse period (20-160 min), and concentration, for 500 min. Ovine ACTH, beta lipotropin, and beta-endorphin immunoreactivity were measured by unextracted RIA. Each pulse of CRF stimulated clearly defined and highly correlated (r greater than 0.9) pulses of the three pituitary hormones, suggesting similar mechanisms controlling release. In dose-response experiments, the minimum concentration of CRF in a 10-min pulse required to significantly raise the output of ACTH was 200 pM, and initial responses had not attained maximal levels with concentrations of CRF increased to 2 microM. Responses to pulsed CRF stimulation decreased with time with all stimulation patterns selected, although previously unstimulated control columns retained the initial capacity to respond. Multiple linear regression analysis showed that hormone output per pulse of CRF (43 nM) increased with increasing pulse period and pulse length. Output of pituitary hormones per unit of CRF applied decreased with pulse length but increased with pulse period. In summary, the responses of proopiomelanocortin derivatives were shown to be sensitive to abrupt increases in CRF, to reduce output under continued stimulation, and to have an inherent time lag before responding maximally to subsequent pulsed stimulation. PMID- 2990877 TI - Putative nuclear triiodothyronine receptors in tadpole liver during metamorphic climax. AB - It has been shown previously that the maximum binding capacity (MBC) of the putative T3 receptors in tadpole red blood cells (RBCs) is increased during development and can be stimulated by treatment with thyroid hormone (TH). The present study was performed to determine if the MBC of tadpole liver nuclei is also increased during development or after treatment with TH. Because of the relatively high levels of endogenous TH in tadpoles during climax, the use of an in vivo saturation assay employing [125I]T3 was not feasible. Thus, MBC was determined by measuring by RIA the amount of T3 bound to the liver nuclei in tadpoles pretreated with sufficient T3 to saturate the receptors. Values were then corrected for the nonsaturable fraction using data obtained in tadpoles given a large dose of T3 (10 nmol). After this dose, essentially all of the T3 in the nucleus was bound to nonsaturable sites. MBC values estimated by this method and by Scatchard analysis were comparable. In contrast to the observations in tadpole RBCs, no significant change in the MBC of liver nuclei occurred as the tadpole progressed from early prometamorphosis to metamorphic climax; in tadpoles at stages XII-XIV and XIX-XXIII, MBC values were 0.308 +/- 0.024 (+/- SE) and 0.260 +/- 0.035 pmol/mg DNA, respectively. Furthermore, treatment of tadpoles with T4 (1 nmol T4; 14 days before study), which resulted in a marked increase in receptor number in RBCs, had no effect on MBC in hepatic nuclei. The amounts of nucleus-bound endogenous T3 in liver and RBCs were also determined. From these data and the MBC values, it was calculated that hepatic and RBC nuclear receptors were, respectively, 80% and more than 90% occupied with T3. These findings indicate that there is tissue specificity in the response of receptor MBC to TH during metamorphosis, and that most of the TH on the receptor during climax is T3. PMID- 2990878 TI - Regulation of the content and phosphorylation of RII by adenosine 3',5' monophosphate, follicle-stimulating hormone, and estradiol in cultured granulosa cells. AB - The mechanisms by which FSH and cAMP induce receptors for LH (RLH) and increase progesterone (P) production in estradiol (E)-primed ovarian granulosa cells remain unclear, but may involve increases in the regulatory subunit of cAMP dependent protein kinase II (RII) and the phosphorylation of specific cellular proteins. To examine the relationship of these events, primary cultures of granulosa cells (10(6) cells/ml) from E-treated (1.5 mg/day for 3 days) immature female rats were incubated with 10 nM E with or without FSH (25 ng/ml) for 0-120 h. The cytosolic content of RII was analyzed by four techniques: 1) immunoblotting using an antibody to bovine heart RII; 2) photoaffinity labeling with [32P]8-azido-cAMP; 3) phosphorylation with [gamma-32P]ATP with or without 2 microM cAMP or with the catalytic subunit of cAMP-dependent protein kinase; and 4) phosphorylation of intact cells with [32P] orthophosphate. All approaches revealed a time-dependent 5- to 6-fold increase in RII content in granulosa cells cultured for 48 h with E and FSH compared to that in cells treated with E alone. The content of RI, the regulatory subunit of protein kinase type I, remained low throughout the culture period regardless of hormone treatment. Granulosa cells were also cultured with E (10 nM) and 8-bromo-cAMP (8-Br-cAMP; 0.25-3 mM) or forskolin (0.5-100 microM), agents that increase intracellular cAMP, for 48 or 72 h. The cytosolic content and phosphorylation of RII were increased by culturing granulosa cells in E and 8-Br-cAMP (1 mM) or forskolin (50 microM) for 48 h. The increase in RII was associated with a FSH-mediated increase in the content and phosphorylation of other cAMP-dependent phosphoproteins. The increases in RII and cAMP-dependent phosphoproteins were associated with specific alterations in granulosa cell function: a FSH-mediated rise in 1) RLH [59.3 +/- 7.4 cpm/micrograms DNA (without FSH) to 1171.5 +/- 157 cpm/micrograms DNA (with FSH]) and 2) P accumulation in the medium [0.05 +/- 0.03 ng/ml (without FSH) to 25.3 +/ 4.6 ng/ml (with FSH]) at 48 h. A dose-dependent increase in the RLH and P accumulation in the medium was observed at 48 h of culture with E and 8-Br-cAMP or E and forskolin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990879 TI - Hyperprolactinemia inhibits gonadotropin-releasing hormone (GnRH) stimulation of the number of pituitary GnRH receptors. AB - Gonadotropin secretion is diminished in the presence of hyperprolactinemia, and previous studies have shown that PRL can reduce GnRH secretion and impair LH responses to GnRH. To investigate the mechanisms of the inhibitory effects of PRL on the pituitary, we administered intraarterial pulse injections of GnRH (25 ng/pulse every 30 min) to castrate testosterone-implanted male rats placed in restraint cages. Serum PRL, GnRH receptor (GnRH-R), and LH responses to GnRH were measured at intervals over 72 h. In control animals which received saline pulses, serum PRL was transiently elevated to the range of 100-150 ng/ml during the first 24 h, GnRH-R remained stable (approximately 300 fmol/mg protein) and serum LH was low (less than 10 ng/ml) throughout the 72 h. GnRH pulses in castrate testosterone-implanted animals increased GnRH-R to values (approximately 600 fmol/mg) similar to those in castrate controls (no testosterone implant, saline pulses) through 48 h, but GnRH-R declined to baseline values by 72 h in both groups. Serum LH responses to GnRH pulses were only present at 24 h. Administration of bromocriptine throughout the 72 h to immobilized castrate rats or to castrate testosterone-replaced animals treated with GnRH pulses suppressed serum PRL, and GnRH-R concentrations remained elevated through 72 h. Serum LH responses to GnRH pulses were 5- to 20-fold higher in bromocriptine-treated rats, and responses were present throughout the 72 h of the experiment. Delaying the start of bromocriptine treatment until 36 h (after the spontaneous PRL peak) resulted in reduced GnRH-R and LH responses at 72 h. Similarly, administration of ovine PRL (during the first 48 h) to bromocriptine-treated rats produced low GnRH R concentrations at 72 h. Thus, the transient elevation of PRL seen in immobilized rats can inhibit the GnRH-stimulated increase in GnRH-R and is associated with reduced LH responses to GnRH. These results indicate that PRL has a direct inhibitory effect on the gonadotrope and suggest that impaired GnRH-R responses to GnRH are one of the mechanisms involved in the diminished gonadotropin secretion seen in hyperprolactinemia. PMID- 2990880 TI - Insulin-like growth factor receptor in fetal lamb liver: characterization and developmental changes. AB - Multiplication-stimulating activity (MSA), an insulin-like growth factor (IGF) (rat IGF II), binds to extracts of many tissues from fetal lambs. We now report the presence of high concentrations of a glycoprotein receptor with a high affinity for MSA-II in microsomes prepared from fetal lamb liver. The binding of radiolabeled MSA-II is inhibited by IGF but not by insulin, human, and ovine GH, ovine PRL, ovine placental lactogen, and mouse epidermal growth factor. The relative potencies of human IGF II, human IGF I, and MSA-II in competing with [125I]MSA-II for binding to this receptor are 100:17:3.5 by weight. Binding is pH, time, and temperature dependent. Gel permeation chromatography of the Triton X-100 soluble receptor indicates a hydrodynamic radius of 6.8 nm. Specific binding increases from mid- to late gestation and is associated with changes in both the affinity and concentration of receptors. Receptor concentration increases from 7.95 +/- 3.94 pmol/mg (mean +/- SE) at 78 days gestation to 15.8 +/- 4.3 pmol/mg at 134-140 days (P less than 0.05), whereas receptor affinity decreases from 1.14 +/- 0.34 X 10(9) liter/mol to 0.63 +/- 0.14 X 10(9) liter/mol over this period (P less than 0.05). The presence of very high concentrations of an IGF receptor in fetal lamb liver suggest that this organ may be a major target for IGF action in fetal life. The increase with advancing gestational age of the concentration of IGF receptors which have preferential specificity for IGF II may function to increase the responsiveness of the fetal lamb liver to IGF II stimulation and so compensate for the decline in plasma concentrations of this growth factor which occur near birth. PMID- 2990881 TI - The use of the corticotropin-releasing hormone test to monitor the recovery of patients with Cushing's disease or Cushing's syndrome due to an adrenal adenoma after adenomectomy. AB - Six patients with Cushing's disease and three with Cushing's syndrome due to an adrenal adenoma were monitored after their adenomectomy with the corticotropin releasing hormone test to evaluate the progress of recovery of their pituitary adrenal function. Before surgery the patients with Cushing's disease showed either high, normal or low responses of plasma ACTH and cortisol to 100 micrograms synthetic ovine corticotropin-releasing hormone (CRH) administered intravenously, whereas all three patients with Cushing's syndrome due to an adrenal adenoma showed no response of plasma ACTH or cortisol to CRH. One or two months after surgery, the patients who had Cushing's disease had low levels of basal plasma ACTH and cortisol and their responses to CRH were extremely low. However, the same patients were tested later, it was found that their responses to CRH gradually increased and reached normal ranges approximately within one year after tumor removal, which coincided with the overall improvement in their clinical signs and symptoms due to adrenal insufficiency. In contrast, the recovery of the pituitary adrenal function in patients who had Cushing's syndrome due to an adrenal adenoma was not complete even one year after surgery. Thus the corticotropin-releasing factor test is a useful criteria to evaluate the recovery of the pituitary adrenal function in these patients after surgery, since the responses of plasma ACTH and cortisol to the administered CRH are parallel with the improvements in clinical signs and symptoms due to adrenal insufficiency in patients with Cushing's disease. PMID- 2990882 TI - Corticoidogenic effect of acetylcholine in bovine adrenocortical cells. AB - The effect of acetylcholine (ACh) on corticoidogenesis in primary cultured bovine adrenocortical cells was examined. One hour exposure to 10(-3) M ACh resulted in a stimulative effect on corticoidogenesis in the freshly isolated cells, and the effect of ACh grew intense during primary culture and reached the maximum on day 2. ACh showed the effect at a higher concentration than 10(-6) M. Thus the primary 2-day cultured cells were used. The corticoidogenic effect of ACh was inhibited by atropine but not by hexamethonium. The effect of ACh was dose dependent, and the extracellular Ca++ was obligatory in inducing the effect. These results suggest that the corticoidogenic effect of ACh may be due to an increase in Ca++-influx via muscarinic receptor in adrenocortical cells. PMID- 2990883 TI - A predicted case with neonatal transient hypothyroidism due to blocking type thyrotropin binding inhibitor immunoglobulins (TBII). AB - A 26-yr-old female with primary hypothyroidism due to potent blocking type thyrotropin binding inhibitor immunoglobulins (TBII) was advised of the high risk of bearing a baby with neonatal transient hypothyroidism. This prediction proved valid and her baby was found to be hypothyroid with a potent TBII. By expressing the baby's TBII as the absolute concentration, we found an almost linear regression of TBII and the half-life was calculated as 18.0 days. The TBII became undetectable by the 6th month and thyroid medication was stopped, however the baby remained euthyroid with subsequent normal physical and mental development. PMID- 2990884 TI - Effect of dexamethasone on the release of pituitary hormones in monolayer cultures of rat pituitary cells. AB - The effect of dexamethasone on the release of ACTH, GH, PRL, LH and TSH was studied in monolayer cultures of rat pituitary cells in 4-hour incubation. With or without the addition of rat hypothalamic extract, the release of GH was significantly inhibited by dexamethasone at concentrations higher than 10(-9) M, although less remarkably than that of ACTH. Intracellular ACTH and GH were unchanged. PRL, LH and TSH were not affected. These results indicate that dexamethasone, when exerted for 4 hours, suppressed the release of GH as well as ACTH, at least in part, at the pituitary level. PMID- 2990885 TI - Bovine thyrotropin inhibits DNA synthesis inversely with stimulation of cyclic AMP production in cultured porcine thyroid follicles. AB - The effects of bovine thyrotropin (TSH) on DNA synthesis and cyclic AMP production were studied in porcine thyroid follicles using suspension culture. During the early 72 hours incubation, the time-dependent uptake of [3H]thymidine by the follicles was observed. In the presence of 10 mU/ml TSH, the uptake of [3H] thymidine was significantly depressed at 72 hours incubation. TSH inhibition of [3H] thymidine incorporation was related to its concentration and the 50% inhibition was observed by using 1.0 mU/ml TSH. Under the same conditions, cyclic AMP production was stimulated by TSH and the stimulation was observed to be related to TSH concentration. In these experiments, the incubation time was 30 min. Dibutyryl cyclic AMP, an analogue of cyclic AMP, inhibited the [3H] thymidine uptake at 72 hours incubation. From these results, it is suggested that TSH inhibits DNA synthesis, and that the inhibition may be mediated by cyclic AMP that is produced by TSH stimulation. PMID- 2990886 TI - Peritoneoscopy as an aid in intravenous injection of indocyanine green (ICG). AB - Peritoneoscopy as an aid in intravenous injection of indocyanine green (ICG) was clinically evaluated. Hepatic parenchyma was stained after intravenous injection of ICG, while interstitial connective tissue, fatty deposition and hepatoma tissue were not. Regenerative hepatic cell mass including dark reddish patchy marking (Shimada's code No. 7) and semispherical areas of regeneration or nodules (Shimada's code No. 8) was well stained and clearly contrasted. There were some cases of chronic active hepatitis, in which liver surface showed spotty staining at sites expected to become regenerative nodules in the future, in contrast to being judged as "no abnormal findings" peritoneoscopically. On the other hand, periportal reddish marking (Shimada's code No. 4) representing piecemeal or bridging hepatic cell necrosis was not stained. PMID- 2990887 TI - Gastroduodenal cytomegalovirus infection after renal transplantation. Fiberscopic observations. AB - Recently we experienced 2 adult post-renal transplantation cases with gastroduodenal cytomegalic inclusion body. In one case, it was possible to carry out close endoscopic observation from onset to resolution a - very rare opportunity. A 39-year-old man developed continuous occult blood in the stool for 1 month after renal transplantation. Multiple erosive lesions from the upper corpus of the stomach to the bulb of the duodenum were revealed by the endoscopic study. A biopsy specimen from the erosion showed the cytomegalic inclusion body. One month later, the erosive lesions had disappeared. The second case is a 40 year-old man who developed tarry stools 5 days after renal transplantation. Multiple erosions partially covered with blood coagula from the upper portion of the gastric corpus to the antrum were revealed by the endoscopic examination. A biopsy specimen from the erosion in the antrum contained the cytomegalic inclusion body. PMID- 2990888 TI - Effects of selenium on quartz-induced cytotoxicity in macrophages. AB - Guinea pig peritoneal macrophages were incubated in vitro with various concentrations of selenium (as sodium selenite), quartz, and quartz + selenite. Cellular viability, adhesiveness, migration, and lipid peroxide content as parameters for cytotoxicity were studied. A dose-dependent cytotoxic response of macrophages to selenite was observed. Selenite concentrations of 10(-6), 10(-5), and 10(-4) M were shown to be nontoxic in the various test procedures. Quartz treatment of macrophages reduced cell viability, adhesiveness, and migration, and enhanced lipid peroxide release. Cotreatment of macrophages with nontoxic selenite concentrations suppressed significantly the cytotoxic effects induced by quartz and was effective in reducing the high rate of lipid peroxidation. The results provide support for the role of selenium in the stabilization of macrophage membrane damaged by quartz. PMID- 2990889 TI - Calculated solubility of chrysotile asbestos in physiological systems. AB - Chrysotile asbestos fibers which are ingested into a physiological system will dissolve in fluids to which they are exposed. The rate of dissolution can be estimated by formulating an appropriate rate law and retrieving rate constants from published experiments. The rate of dissolution calculated for asbestos fibers is dependent on the surface area of dissolving mineral, mass of solution to which the fibers are exposed within cells, the rate of transport of H+ into the cells, and the rate of transport of Mg2+ and SiO2 out of the cells. The maximum rate of dissolution of a single 10-micron X 1-micron fiber within a cell is 2.13 X 10(-7) moles hr-1 kg-1 consistent with dissolution rates measured in vivo by Morgan et al. (A. Morgan, A. Holmes, and C. Gold. Studies of the solubility of constituents of chrysotile asbestos in vivo using radioactive tracer techniques. Environ. Res. 4, 558-570, 1971). PMID- 2990890 TI - Psychosocial enhancement of immunocompetence in a geriatric population. AB - This study assessed the enhancement of immunocompetence by relaxation and social contact in 45 geriatric residents of independent-living facilities. Subjects were randomly assigned to one of three protocols: relaxation training, social contact, or no contact. Subjects in the relaxation and social-contact conditions were seen individually three times a week for a month. Blood samples and self-report data were obtained at baseline, at the end of the intervention, and at a 1-month follow-up. At the end of the intervention, the relaxation group showed a significant increase in natural killer cell activity, and significant decreases in antibody titers to Herpes simplex virus and self-rated distress; the other two groups showed nonsignificant changes. There was a general increase in the T lymphocyte response to phytohemagglutinin stimulation at the end of the intervention, with greater change at lower mitogen concentrations. These data suggest that cellular immunocompetence may be enhanced by psychosocial interventions. PMID- 2990892 TI - The cytoplasmically-made subunit IV is necessary for assembly of cytochrome c oxidase in yeast. AB - Yeast cytochrome c oxidase contains three large subunits made in mitochondria and at least six smaller subunits made in the cytoplasm. There is evidence that the catalytic centers (heme a and copper) are associated with the mitochondrially made subunits, but the role of the cytoplasmically-made subunits has remained open. Using a gene interruption technique, we have now constructed a Saccharomyces cerevisiae mutant which lacks the largest of the cytoplasmically made subunits (subunit IV). This mutant is devoid of cyanide-sensitive respiration, the absorption spectrum of cytochrome aa3 and cytochrome c oxidase activity. It still contains the other cytochrome c oxidase subunits but these are not assembled into a stable complex. Active cytochrome c oxidase was restored to the mutant by introducing a plasmid-borne wild-type subunit IV gene; no restoration was seen with a gene carrying an internal deletion corresponding to amino acid residues 28-66 of the mature subunit. Subunit IV is thus necessary for proper assembly of cytochrome c oxidase. PMID- 2990891 TI - Effects of inhibitors of glycoprotein processing on the synthesis and biological activity of the erb B oncogene. AB - Three glycoprotein-processing inhibitors were used to resolve whether correct glycosylation was required for the oncogenic activity of erb B. The two glucosidase-I inhibitors, 1-deoxynojirimycin and 2,5-dihydroxymethyl 3,4 dihydroxypyrrolidine, arrested processing of v-erb B at the immature 68-kd form whereas, in the presence of the alpha-mannosidase-II inhibitor (swainsonine), cells synthesised an abnormally processed 70-kd form of v-erb B. Transport of incorrectly processed v-erb B to the cell surface was, however, unaffected, suggesting that correct processing is not a prerequisite for intracellular routing of v-erb B. Two systems were used to assess whether incorrectly processed erb B could maintain the transformed state. The first asked whether inhibitor treatment would release temperature-sensitive avian erythroblastosis virus (AEV) transformed erythroblasts kept at the viral permissive temperature from the erb B induced block in differentiation, as seen when cells are normally shifted to the non-permissive temperature. The second tested the ability of AEV-transformed fibroblasts to grow in soft agar. In both systems, all three processing inhibitors did not alter the transformed phenotype suggesting that correct carbohydrate processing is not required for the transforming activity of erb B. In addition, none of the three processing inhibitors were found to have any effect on the normal maturation of bone marrow CFU-E or induced differentiation of temperature-sensitive AEV-transformed erythroblasts. PMID- 2990893 TI - RNA processing in Neurospora crassa mitochondria: use of transfer RNA sequences as signals. AB - We have used RNA gel transfer hybridization, S1 nuclease mapping and primer extension to analyze transcripts derived from several genes in Neurospora crassa mitochondria. The transcripts studied include those for cytochrome oxidase subunit III, 17S rRNA and an unidentified open reading frame. In all three cases, initial transcripts are long, include tRNA sequences, and are subsequently processed to generate the mature RNAs. We find that endpoints of the most abundant transcripts generally coincide with those of tRNA sequences. We therefore conclude that tRNA sequences in long transcripts act as primary signals for RNA processing in N. crassa mitochondria. The situation is somewhat analogous to that observed in mammalian mitochondrial systems. The difference, however, is that in mammalian mitochondria, noncoding spacers between tRNA, rRNA and protein genes are very short and in many cases non-existent, allowing no room for intergenic RNA processing signals whereas, in N. crassa mtDNA, intergenic non coding sequences are usually several hundred nucleotides long and contain highly conserved GC-rich palindromic sequences. Since these GC-rich palindromic sequences are retained in the processed mature RNAs, we conclude that they do not serve as signals for RNA processing. PMID- 2990894 TI - Amber, ochre and opal suppressor tRNA genes derived from a human serine tRNA gene. AB - Amber, ochre and opal suppressor tRNA genes have been generated by using oligonucleotide directed site-specific mutagenesis to change one or two nucleotides in a human serine tRNA gene. The amber and ochre suppressor (Su+) tRNA genes are efficiently expressed in CV-1 cells when introduced as part of a SV40 recombinant. The expressed amber and ochre Su+ tRNAs are functional as suppressors as demonstrated by readthrough of the amber codon which terminates the NS1 gene of an influenza virus or the ochre codon which terminates the hexon gene of adenovirus, respectively. Interestingly, several attempts to obtain the equivalent virus stock of an SV40 recombinant containing the opal suppressor tRNA gene yielded virus lacking the opal suppressor tRNA gene. This suggests that expression of an efficient opal suppressor derived from a human serine tRNA gene is highly detrimental to either cellular or viral processes. PMID- 2990895 TI - Co-transcribed 3' host sequences augment expression of integrated hepatitis B virus DNA. AB - We have previously reported the cloning and structural analysis of integrated hepatitis B virus DNA copies from the human hepatoma cell line PLC/PRF/5. Here we show that the cloned DNA fragments of 10.7 kb and 10.5 kb contain intact coding sequences for HBsAg since Ltk- cells transfected with these DNAs secrete considerable amounts of HBsAg. We show for the 10.7-kb fragment that multiple readthrough messages composed of viral as well as cellular sequences are transcribed. These RNAs differ only in their 3' sequences. Furthermore, the 10.7 kb insert leads to a substantial increase in HBsAg produced compared with HBV DNA and with the 10.5-kb insert. We provide evidence that the different 3' sequences on the HBsAg transcripts account for the augmentation of expression. PMID- 2990896 TI - Oscillations and cyclic AMP-induced changes of the K+ concentration in Dictyostelium discoideum. AB - By means of a K+-sensitive electrode, the extracellular K+ concentration was monitored in cell suspensions of Dictyostelium discoideum. In aggregative cells the attractant cyclic AMP induced a transient release of K+. The response was detectable within 6-12 s and peaked at 30-40 s. The apparent rate of release amounted to 7 X 10(8)K+ ions per cell per min. Adenosine and 5' AMP, which are chemotactically inactive, did not elicit measurable K+ responses. The cyclic AMP induced release of K+ depended on the state of differentiation of the cells. In undifferentiated cells cyclic AMP did not cause a measurable K+ release, whereas folic acid, a potent attractant at this cell stage, induced a weak but significant K+ response. The cyclic AMP-induced K+ release in aggregative cells was inhibited by K+-channel blockers such as quinine and tetraethylammonium. In suspensions of differentiated cells free running oscillations of the extracellular K+ concentration were observed. K+ oscillations were related to cyclic AMP oscillations and oscillations of the light-scattering properties of cells. Cells continuously released NH4+; however, cyclic AMP did not induce a measurable change of NH4+ release. PMID- 2990897 TI - Lines of BPV-transformed murine cells that constitutively express influenza virus hemagglutinin. AB - We have developed and characterized several murine cell lines that constitutively express either the full-length, membrane-bound form of influenza virus hemagglutinin (HA) or a truncated version of the protein (HAsec) that lacks the carboxyterminal anchoring sequences and is secreted from cells. cDNAs encoding HA or HAsec were linked to the murine metallothionein-I promoter or the SV40 early promoter, and inserted into plasmids containing the transforming DNA fragment of bovine papilloma virus (BPV). The resulting vectors were introduced into three cultured lines of murine cells--C127, NIH3T3 and MME--either alone or in the presence of a plasmid that carries the aminoglycoside transferase gene of Tn5. The resulting lines of MME cells contained 1-5 copies of the vector in an integrated state and expressed low levels of HA (approximately 10(4) molecules/cell). In contrast, lines of C127 and NIH3T3 cells were obtained that express up to 5 X 10(6) molecules of HA per cell or secrete approximately 10(7) molecules of HAsec per cell per 24 h. Some of these cell lines carry multiple (30 200) copies of the vector in an integrated state; in others, the vector is propagated as unit-length episomes or as oligomers. Both the membrane-bound and secreted forms of HA expressed in these cell lines display a more extensive pattern of glycosylation than HA or HAsec synthesized in simian cells and they are transported to the cell surface more slowly. Pulse-chase experiments suggest that the step which limits the rate at which HA and HAsec travel down the secretory pathway occurs in the rough endoplasmic reticulum before the molecules are transferred to the Golgi apparatus. Using indirect immunofluorescence in combination with a cell sorter, we have shown that the level of expression of HA within cloned populations of producing cells can be variable. However, greater than 90% of the cells in certain cell lines display considerable quantities of HA on their surface, as judged by their ability to bind red blood cells in large numbers. We have taken advantage of the membrane fusion activity of HA to effect the fusion of erythrocytes to these cells and to deliver the contents of red cell ghosts into the cells' cytoplasm. PMID- 2990898 TI - An enzymatic assay reveals that proteins destined for the apical or basolateral domains of an epithelial cell line share the same late Golgi compartments. AB - The expression of viral envelope proteins on the plasma membrane domains of the epithelial cell line, MDCK, is polar. Influenza virus infection of these cells leads to expression of the viral haemagglutinin and neuraminidase glycoproteins on the apical domain of the plasma membrane while vesicular stomatitis virus (VSV) infection yields basolateral expression of the sialic acid-bearing G protein. We have exploited the ability of the influenza neuraminidase to desialate the G protein of VSV to test for contact between these proteins during their intracellular transport to separate plasma membrane domains. We were able to select for VSV-G protein expression in doubly-infected cells because VSV protein production was accelerated in cells pre-infected with influenza virus. During double infection the envelope proteins of both viruses displayed the same polar localization as during single infection but the VSG-G protein was undersialated due to the action of the influenza neuraminidase. Incubation of singly-infected cells at 20 degrees C blocked the transport of VSV-G protein to the cell surface and resulted in increased sialation of the protein over that seen at 37 degrees C. This suggests that G protein is held in contact with the sialyl transferase at this temperature. 20 degrees C incubations of doubly infected cells also produced the undersialated G protein characteristic of interaction with the neuraminidase. We conclude that most of the newly synthesised basolaterally-directed G protein is in physical contact with the majority of the neuraminidase through the terminal steps of Golgi processing. PMID- 2990899 TI - The chromatin structure of Rous sarcoma proviruses is changed by factors that act in trans in cell hybrids. AB - In several lines of Rous sarcoma virus (RSV)-transformed rat cells the proviruses are in a configuration typical of active eukaryotic genes. They are sensitive to pancreatic DNase I, with sites hypersensitive to nuclease near the 5' end of the genome, they are close to the nuclear 'cage' and they show a low level of cytosine methylation in CpG doublets. In contrast, in phenotypically untransformed hybrids between these cells and uninfected rat or mouse cells, RSV inactivity is associated with hypermethylation of the provirus, reduced DNase I sensitivity (in two out of three examples) and, where examined, relative remoteness from the nuclear cage. These changes in proviral configuration, which occur rarely in spontaneous reversion of transformed cells, can thus be induced at high frequency and stability in cell hybrids by trans-acting influences of the uninfected parents. PMID- 2990900 TI - In vivo evidence that the nusA and infB genes of E. coli are part of the same multi-gene operon which encodes at least four proteins. AB - Previous work has shown that the Escherichia coli nusA gene codes for a protein which regulates transcription termination. The 16.0-kb EcoRI DNA fragment that includes the nusA gene, codes for at least eight bacterial proteins of mol. wts. 48 000 (argG), 21 000 (p21), 64 000 (nusA), 120 000 (IF2 alpha)-(infB), 91 000 (IF2 beta)(infB), 15 000 (p15), 10 000 (rpsO) and 85 000 (pnp). We have constructed several deletion and fusion derivatives from this cloned DNA and examined in vivo the structure and expression of these genes. First, the promoter functional in vivo for the nusA gene was mapped at approximately 800 bp upstream of the nusA structural gene. Second, the synthesis of five proteins, p21, NusA, IF2 alpha, IF2 beta (and p15) proteins, was affected by the deletion of the nusA promoter. Third, these same five proteins were hyperproduced after fusion of the DNA fragment to the lambda pL promoter. In addition, subcloning experiments revealed that the p15 gene is expressed by the read-through transcription from the infB gene. These results lead us to conclude that the genes coding for the p21, NusA, InfB (IF2 alpha and IF2 beta), and p15 proteins form a single transcriptional unit ('nusA-infB operon') in vivo and that rpsO and pnp genes do not belong to the same operon. The in vivo attenuation site of this operon is described. PMID- 2990901 TI - Cloning and expression of the fbc operon encoding the FeS protein, cytochrome b and cytochrome c1 from the Rhodopseudomonas sphaeroides b/c1 complex. AB - The gene for the FeS protein of the Rhodopseudomonas sphaeroides b/c1 complex was identified by means of cross-hybridization with a segment of the gene encoding the corresponding FeS protein of Neurospora crassa. Plasmids (pRSF1-14) containing the cross-hybridizing region, covering in total 13.5 kb of chromosomal DNA, were expressed in vitro in a homologous system. One RSF plasmid directed the synthesis of all three main polypeptides of the R. sphaeroides b/c1 complex: the FeS protein, cytochrome b and cytochrome c1. The FeS protein and cytochrome c1 were apparently synthesized as precursor forms. None of the pRSF plasmids directed the synthesis of the 10-kd polypeptide found in b/c1 complex preparations. Partial sequencing of the cloned region was performed. Several sites of strong homology between R. sphaeroides and eukaryotic polypeptides of the b/c1 complex were identified. The genes encode the three b/c1 polypeptides in the order: (5') FeS protein, cytochrome b, cytochrome c1. The three genes are transcribed to give a polycistronic mRNA of 2.9 kb. This transcriptional unit has been designated the fbc operon; its coding capacity corresponds to the size of the polycistronic mRNA assuming that only the genes for the FeS protein (fbcF), cytochrome b (fbcB) and cytochrome c1 (fbcC) are present. This could indicate that these three subunits constitute the minimal catalytic unit of the b/c1 complex from photosynthetic membranes. PMID- 2990902 TI - dnaA protein-regulated transcription: effects on the in vitro replication of Escherichia coli minichromosomes. AB - Both initiation of replication and initiation of transcription are influenced by dnaA protein, when minichromosomes are assayed in vitro for dnaA protein complementation. This dnaA protein effect is seen only if minichromosomes are used containing the 16-kd promoter, from which transcription is directed into the minimal origin. Determination of the 16-kd promoter activity both in vivo and in vitro showed that this strong promoter is specifically repressed by dnaA protein. The 16-kd promoter is thus an integral regulatory region of oriC. PMID- 2990903 TI - Does unpaired adenosine-66 from helix II of Escherichia coli 5S RNA bind to protein L18? AB - Adenosine-66 is unpaired within helix II of Escherichia coli 5S RNA and lies in the binding site of ribosomal protein L18. It has been proposed as a recognition site for protein L18. We have investigated further the structural importance of this nucleotide by deleting it. The 5S RNA gene of the rrnB operon of E. coli was subjected to primer-directed mutagenesis. To produce the deletion it was necessary to use simultaneously the mutagenic dodecamer dCGGCGCACGGCG and the universal M13 primer dCCCAGTCACGACGTT, and to employ forced annealing conditions. The mutated gene was expressed in an overproducing plasmid derived from pKK3535. Binding studies with protein L18 revealed that the protein bound much more weakly to the mutated 5S RNA. We consider the most likely explanation of this result is that L18 interacts with adenosine-66, and we present a tentative model for an interaction between the unpaired adenosine and the adjacent guanosine-67 of the RNA and glutamine-19 of the protein L18. PMID- 2990904 TI - Negative supercoiling induces spontaneous unwinding of a bacterial promoter. AB - We have examined the influence of negative supercoiling on the DNA structure of a bacterial promoter (tyrT from Escherichia coli), the transcriptional activity of which is strongly enhanced by torsional stress in vitro. Certain regions of this promoter become sensitive to digestion by single-strand-specific S1 nuclease as a consequence of negative superhelicity. These regions occur with high frequency (1 per 30-50 bp) and are normally centered on a TpA doublet. The major positions of cleavage are located in and around the -10 sequence TATGATG, the unwinding of which is a prerequisite for gene expression. An apparently trivial change in the 10 sequence from TATGATG to TATGAAG reduces both transcriptional activity and S1 nuclease sensitivity at least 10-fold. Thus the nuclease sensitivity of the promoter correlates strongly with its biological function; and both of these phenomena correlate with certain sequence-dependent structural properties of the DNA. PMID- 2990905 TI - Failure of E. coli K-12 to transport PhoE-LacZ hybrid proteins out of the cytoplasm. AB - A phoE-lacZ hybrid gene encoding the N-terminal 300 amino acid residues of pre PhoE protein, fused to an almost complete beta-galactosidase molecule was constructed in vitro. Cell fractionation experiments suggested that the hybrid gene product is transported to the outer membrane. However, by using immuno cytochemical labelling on ultra-thin cryosections it was shown that the hybrid protein accumulated in the cytoplasm. Thus, it appears that: (i) data on the localization of hybrid proteins merely based on cell fractionation experiments are not reliable, and (ii) either the C-terminal 15% of PhoE protein contain information which is essential for transport, or PhoE-LacZ hybrid proteins can never be transported out of the cytoplasm. The implications of these results for current models on the translocation of outer membrane proteins are discussed. PMID- 2990906 TI - Regulation of the expression of the tufB operon: DNA sequences directly involved in the stringent control. AB - We have located the DNA sequence involved in the stringent control of the Escherichia coli tufB operon. Various deletion and insertion mutants of the promoter locus were constructed by in vitro mutagenesis, and their response to guanosine-5'-diphosphate-3'-diphosphate (ppGpp) was examined in a cell-free transcription system consisting of purified RNA polymerase holoenzyme. The nucleotide sequence (GpCpGpC) from positions -7 to -4 (designating the initiation site of mRNA as position +1) is responsible for the selective inhibition by ppGpp of tufB transcription. Point mutations were then constructed in which each one of the above four nucleotides was replaced by an A or T residue and tested for their response to ppGpp in the in vitro transcription system. The results indicated that the alteration of any nucleotide in the GpCpGpC sequence leads to the loss of the stringent response. PMID- 2990907 TI - Sequence of the relB transcription unit from Escherichia coli and identification of the relB gene. AB - Escherichia coli relB mutants react to amino acid starvation by several abnormal responses, including accumulation of a translational inhibitor. We have isolated a relB-complementing plasmid from the Clarke and Carbon E. coli DNA library. From this plasmid we sequenced a 2140-bp segment which included the relB gene by the following two criteria: (i) it complements chromosomal relB mutations, (ii) the corresponding DNA segment cloned from chromosomal DNA of three relB mutants was defective in relB complementation. All three mutations fell within an open reading frame of 79 amino acids. A polypeptide of 9 kd compatible with this open reading frame was synthesized in maxicells and is in all probability the product of the relB gene. By nuclease S1 mapping we have determined the transcription start and stop of an 870 base transcript of the relB gene. PMID- 2990908 TI - Immobilization and purification of enzymes with staphylococcal protein A gene fusion vectors. AB - Two improved plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusions, have been constructed. These vectors allow fusion of any gene to the protein A moiety, giving fusion proteins which can be purified, in a one-step procedure by IgG affinity chromatography. One vector, pRIT2, is designed for temperature-inducible expression of intracellular fusion proteins in Escherichia coli and the other pRIT5, is a shuttle vector designed for secretion. The latter gives a periplasmatic fusion protein in E. coli and an extracellular protein in Gram-positive hosts such as Staphylococcus aureus. The usefulness of these vectors is exemplified by fusion of the protein A gene and the E. coli genes encoding the enzymes beta-galactosidase and alkaline phosphatase. High amounts of intact fusion protein are produced which can be immobilized on IgG-Sepharose in high yield (95-100%) without loss of enzymatic activity. Efficient secretion in both E. coli and S. aureus, was obtained for the alkaline phosphatase hybrid, in contrast to beta-galactosidase which was only expressed efficiently using the intracellular system. More than 80% of the protein A alkaline-phosphatase hybrid protein can be eluted from IgG affinity columns without loss of enzymatic activity. PMID- 2990909 TI - Immunoelectron microscopic localization of acidic intracellular compartments in hepatoma cells. AB - Using protein A-colloidal gold immunoelectron microscopy and monospecific antibodies to the weak base primaquine, we have delineated acidic intracellular compartments in the human hepatoma cell line, HepG2. Primaquine specifically accumulated within endocytotic compartments (including CURL vesicles, multivesicular bodies and lysosomes). In addition, the Golgi cisternae were positive. However, the CURL tubules, which contain recycling asialoglycoprotein receptor, did not accumulate primaquine. Thus, there may be a gradient of acidification within the endocytotic pathway. PMID- 2990910 TI - A deletion adjacent to the maize transposable element Mu-1 accompanies loss of Adh1 expression. AB - Insertion of the maize transposable element Mu-1 into the first intron of the alcohol dehydrogenase locus (Adh1) of maize produced mutant Adh1-S3034 with 40% of the wild-type level of protein and mRNA. Continued instability at this locus resulted in secondary mutations with lower levels of protein expression. One of these, Adh1-S3034a, has no detectable ADH1 expression. This paper describes the precise nature of the changes in the Adh1 gene that gave rise to the S3034a allele. The Mu-1 element is still present in the mutant, but Adh1 sequences immediately adjacent to the element are deleted. The deletion starts precisely at the Mu-1 insertion site and extends 74 bp leftward removing part of the first intron, the intron:exon junction and 2 bp of the eleventh amino acid codon in the first exon of the gene. Tests for reversion within the somatic tissue of plants show that mutant S3034a, unlike its progenitor, is stably null for ADH1 activity. PMID- 2990911 TI - Characterization of two Phaseolus vulgaris phytohemagglutinin genes closely linked on the chromosome. AB - A lambda 1059 library of Phaseolus vulgaris cv. 'Tendergreen' DNA was screened with a cloned lectin-like cDNA. Among the phages selected was clone lambda B10 containing two complete lectin genes in the same orientation approximately 4 kb apart. The DNA sequences of the lectin genes and their flanking regions have been determined and their transcriptional initiation sites were located by S1 nuclease mapping. On the basis of the deduced amino acid sequences and compositions and the mol. wts. of their encoded glycoproteins, the genes, dlec1 and dlec2, are predicted to encode erythro- and leucoagglutinating phytohemagglutinins (PHA-E and PHA-L), respectively. The mRNA coding regions of dlec1 and dlec2 are 90% homologous, suggesting an origin involving duplication of an ancestral gene. Both lectin genes are intronless and have at least two ATG codons in a short (11-14 bp) 5'-untranslated region. Most of their 5'-untranslated regions consist of alternating pyrimidines and purines (RY repeats). Upstream sequences are also highly conserved between dlec1 and dlec2, including stretches of nine or more alternating R and Y residues. RY repeats of such length are not found within the protein coding portion of dlec1, dlec2 or a Phaseolus lectinlike gene previously described. Overlapping double (dlec1) or triple (dlec2) polyadenylation addition signals are found and there is an unusually high degree of homology (84%) between their 3'-untranslated regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990913 TI - Transcription occurs at a nucleoskeleton. AB - Native chromatin aggregates under isotonic conditions so it is generally studied using higher or lower salt concentrations. This has led to different interpretations of how transcription might occur. Studies using hypertonically isolated preparations suggest that DNA functions in close association with a skeletal nuclear substructure, the matrix or cage, but such a structure is not usually seen under hypotonic conditions (e.g., in 'Miller-spreads'). Using a novel method for preparing chromatin under isotonic conditions we have investigated the site of transcription. We find that all three constituents of the transcription complex, nascent transcripts, active RNA polymerase and genes being transcribed are all closely associated with some structure too large to be electroeluted from the nucleus. Hypotonic treatment partly disrupts this association. We suggest a model for transcription that involves the participation of a nucleoskeleton at the active site and reconcile the contradictory results obtained using different salt concentrations. PMID- 2990912 TI - A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. AB - The construction and use of a Tn3-lac transposon, Tn3-HoHo1, is described. Tn3 HoHo1 can serve as a transposon mutagen and provides a new and useful system for the random generation of both transcriptional and translational lacZ gene fusions. In these fusions the production of beta-galactosidase, the lacZ gene product, is placed under the control of the gene into which Tn3-HoHo1 has inserted. The expression of the gene can thus be analyzed by monitoring beta galactosidase activity. Tn3-HoHo1 carries a non-functional transposase gene; consequently, it can transpose only if transposase activity is supplied in trans, and is stable in the absence of this activity. A system for the insertion of Tn3 HoHo1 into sequences specifically contained within plasmids is described. The applicability of Tn3-HoHo1 was demonstrated studying three functional regions of the Agrobacterium tumefaciens A6 Ti plasmid. These regions code for octopine catabolism, virulence and plant tumor phenotype. The regulated expression of genes contained within each of these regions was analyzed in Agrobacterium employing Tn3-HoHo1 generated lac fusions. PMID- 2990914 TI - Differential expression of human globin genes introduced in K562 cells. AB - We have constructed a series of hybrid globin genes between the 5' half of the human epsilon-, gamma- and beta-globin genes, and the 3' half of the rabbit beta globin gene. These hybrid genes were introduced into the cell line K562. Analysis of the hybrid mRNA populations show that only the hybrid epsilon- and gamma globin genes are expressed in these cells in concordance with the endogenous K562 globin gene expression. It is therefore likely that these cells contain transcriptional factors which allow the specific expression of introduced epsilon and gamma-, but not beta-globin genes, irrespective of their chromosomal localization. PMID- 2990915 TI - Specific binding of radioiodinated granulocyte-macrophage colony-stimulating factor to hemopoietic cells. AB - The hemopoietic growth factor granulocyte-macrophage colony-stimulating factor, GM-CSF, specifically controls the production of granulocytes and macrophages. This report describes the binding of biologically-active 125I-labeled murine GM CSF to a range of hemopoietic cells. Specific binding was restricted to murine cells and neither rat nor human bone marrow cells appeared to have surface receptors for 125I-labeled GM-CSF. 125I-Labeled GM-CSF only appeared to bind specifically to cells in the myelomonocytic lineage. The binding of 125I-labeled GM-CSF to both bone marrow cells and WEHI-3B(D+) was rapid (50% maximum binding was attained within 5 min at both 20 degrees C and 37 degrees C). Unlabeled GM CSF was the only polypeptide hormone which completely inhibited the binding of 125I-labeled GM-CSF to bone marrow cells, however, multi-CSF (also called IL-3) and G-CSF partially reduced the binding of 125I-labeled GM-CSF to bone marrow cells. Interestingly, the binding of 125I-labeled GM-CSF to a myelomonocytic cell line, WEHI-3B(D+), was inhibited by unlabeled GM-CSF but not by multi-CSF or G CSF. Scatchard analysis of the binding of 125I-labeled GM-CSF to WEHI-3B(D+) cells, bone marrow cells and peritoneal neutrophils indicated that there were two classes of binding sites: one of high affinity (Kd1 = 20 pM) and one of low affinity (Kd2 = 0.8-1.2 nM). Multi-CSF only inhibited the binding of 125I-labeled GM-CSF to the high affinity receptor on bone marrow cells: this inhibition appeared to be a result of down regulation or modification of the GM-CSF receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990917 TI - Inactivation and reactivation of a variant-specific antigen gene in cyclically transmitted Trypanosoma brucei. AB - In Trypanosoma brucei, the activation of the variant-specific antigen gene AnTat 1.1 proceeds by the synthesis of an additional gene copy, the AnTat 1.1 ELC, which is transposed to a new location, the expression site, where it is transcribed. Using the AnTat 1.1 variant to infect flies, we investigated the fate of the AnTat 1.1 ELC during cyclic transmission of T. brucei. We show here that the AnTat 1.1 ELC is conserved in procyclic trypanosomes, obtained either from the midgut of infected Glossina or from cultures, and in metacyclic trypanosomes, although the AnTat 1.1 serotype is not detected among metacyclic antigen types. This same AnTat 1.1 ELC, which is thus silent as the parasite develops in the insect vector, can be reactivated without duplication during the first parasitemia wave following cyclical transmission. This re-expression of the conserved ELC accounts for the early appearance of the 'ingested' antigenic type after passage through the fly. PMID- 2990916 TI - Enhancer activity correlates with the oncogenic potential of avian retroviruses. AB - Avian retroviruses lacking an oncogene, such as Rous-associated virus 1 (RAV-1), RAV-2, and td mutants of Rous sarcoma virus (RSV), can nevertheless cause leukemias and other neoplastic diseases. During this process, viral DNA integrates near a cellular proto-oncogene, such as c-myc, and thus de-regulates its expression. The virus RAV-0, on the other hand, is known to be non-oncogenic even in long-term in vivo infections of domestic chickens. The major difference between oncogenic and non-oncogenic viruses is found within the U3 region of the long terminal repeat (LTR) which is known to harbor the promoter and enhancer elements. We therefore wanted to see whether viral oncogenicity was correlated with enhancer activity. Using a variety of techniques (including the SV40 'enhancer trap' from which we obtained RSV-SV40 recombinant viruses), we demonstrate that a strong enhancer exists within the LTRs of both RSV and RAV-1. In contrast, no enhancer is present in RAV-0, although RAV-0 has functional promoter elements. Our data therefore strongly support a concept of oncogenesis by enhancer insertion. PMID- 2990918 TI - A developmentally regulated cysteine proteinase in Dictyostelium discoideum. AB - We have determined the sequence of a Dictyostelium mRNA encoding a protein with a high degree of homology to plant and animal cysteine proteinases. The degree of homology is highest in the region of the cysteine residue which is transiently acylated during peptide hydrolysis but all other residues known to be important in catalysis are also conserved. We have named this protein cysteine proteinase 1. There is a hydrophobic signal peptide of 18 amino acids and an additional 99 amino acids at the N terminus, which are not present in other cysteine proteases and which may be cleaved off during processing of the enzyme. There is a single copy of the gene in the Dictyostelium genome. The cysteine proteinase 1 mRNA is absent from growing cells and from cells isolated during the first 6 h of development but it constitutes approximately 1% of cellular mRNA by 10-12 h of development. During the development of Dictyostelium a major fraction of cellular protein is degraded to provide amino acids and a source of energy. Cysteine proteinase 1 may play a role in this auto-digestion. PMID- 2990919 TI - Analysis of the interferon system in African patients with acquired immunodeficiency syndrome. AB - Serum interferon and in vitro production of alpha and gamma interferon by peripheral blood leucocytes were examined in 21 African patients with acquired immunodeficiency syndrome (AIDS) and in 15 African patients with AIDS-related complex. Interferon was detected in the serum of 44% of the patients with AIDS related complex and in 70% of the patients with full-blown AIDS, and was characterized as an acid-labile alpha interferon. When compared to healthy blood donors, the interferon response of peripheral blood leucocytes to Newcastle Disease virus was impaired in 7 of 12 patients with AIDS-related complex and in 16 of 20 AIDS patients (p less than 0.005). Also, production of gamma interferon following stimulation with phytohaemagglutinin was diminished in 5 of 11 patients with AIDS-related complex and in 13 of 17 patients with AIDS (p less than 0.005). A high correlation was observed between the presence of circulating interferon and decreased in vitro production of gamma interferon, but not of alpha interferon. These results suggest that the impairment of in vitro production of gamma interferon can be used as a preclinical marker of AIDS. PMID- 2990920 TI - Method for selecting optimal cells for enterovirus isolation as determined in an outbreak of echovirus type 33 meningitis. AB - An outbreak of echovirus type 33-induced meningitis which occurred in Belgium in 1982 is reported. To identify the causative agent, titers of an early isolate were measured on a variety of cells in order to select the optimal cells. A human rhabdomyosarcoma cell line was found to be the substrate of choice due to its proficient isolation of the virus. This method of determining infectious titres is recommended for improving enterovirus isolation in other epidemics. PMID- 2990922 TI - A comparison of the structural requirements for DNA cleavage by the isoschizomers HaeIII, BspRI and BsuRI. AB - We have investigated the structural requirements for DNA cleavage by the isoschizomers HaeIII, BspRI and BsuRI which recognize the sequence -d(GGCC)-. For this purpose decadeoxynucleotides were synthesized by the solid-phase phosphotriester method and purified by high-performance liquid chromatography. The kinetics of cleavage of these oligodeoxynucleotides were determined for the three isoschizomers with the following results. The sequence adjacent to the recognition site strongly influences the rate of cleavage. The preference is qualitatively the same for all three enzymes: AGGCCT greater than TGGCCA greater than GGGCCC approximately equal to CGGCCG, and follows the thermal stability of the different decanucleotides. Substitutions within the recognition site, namely dI for dG and dU for dC, affect the rate of cleavage differently for the three enzymes. The results can be rationalized in terms of an interaction of HaeIII with the major and minor groove of the DNA, of BspRI mainly with the minor groove and of BsuRI with the major groove of DNA. It is obvious from our data that the mechanism of recognition of the same site is different for the three isoschizomers. PMID- 2990921 TI - Detection of active Epstein-Barr infection in pregnant women. AB - Sera taken from pregnant patients and their newborns at delivery were examined for evidence of primary or reactivated Epstein-Barr virus infection. Of 102 women, 37 showed serological signs of reactivated and two signs of primary infection. A mild congenital defect was observed in association with one of the two cases of maternal primary infection. Infants of mothers with reactivated infections remained healthy during the one year follow-up after birth. PMID- 2990923 TI - Investigation of the solution structure of a DNA octamer [d(GGAATTCC)]2 using two dimensional nuclear Overhauser enhancement spectroscopy. AB - Proton two-dimensional nuclear Overhauser enhancement (2D NOE) spectra in the pure absorption phase were obtained at 500 MHz for [d(GGAATTCC)]2 in aqueous solution at a series of mixing times. The experimental data were analyzed by comparison with theoretical spectra calculated using the complete 70 X 70 relaxation matrix including all proton dipole-dipole interactions and spin diffusion [Keepers, J. W. & James, T. L. (1984) J. Magn. Reson. 57, 404-426]. The theoretical spectra at each mixing time were calculated using two structures: a standard B-form DNA structure and an energy-minimized structure based on the similarity of the six internal residues of the title octamer with those of the dodecamer [d(CGCGAATTCGCG)]2, for which the crystal structure has been determined. Neither the standard B-form nor the energy-minimized structure will yield theoretical 2D NOE spectra which accurately reproduce all peak intensities in the experimental spectra. However, many features of the experimental spectra can be represented by both the B-form and the energy-minimized structure. Sequence-dependent structural characteristics are manifest in the 2D NOE spectra, in particular at the purine-pyrimidine junction as noted previously in the crystal structure. On the whole, the energy-minimized structure appears to yield theoretical 2D NOE spectra which mimic many, if not all, aspects of the experimental spectra. All 2D NOE data were consistent with nanosecond correction times as implied by proton spin-lattice relaxation time measurements. But better fits of some of the 2D NOE data using small variations in an effective isotropic correlation time suggest that there may be some local variations in mobility within the octamer duplex structure in solution. PMID- 2990924 TI - Isolation and biochemical characterization of maleic-acid hydratase, an iron requiring hydro-lyase. AB - A procedure for the isolation of maleic acid hydratase (D-malate hydro-lyase, EC 4.2.1.31) of about 95% purity from rabbit kidneys is described. The enzyme consists of a single polypeptide chain of 582 amino-acid residues with an approximate molecular mass of 68 kDa. The enzyme is very unstable and has an absolute requirement for chloride ions. Addition of sodium sulphide during the purification process was essential to maintain the enzyme in an activatable state. The pure preparation has low activity but responds to activation with Fe2+ ions, Na2S and a thiol. The sequence of adding the activating reagents is critical to achieve optimal activity. Ni2+ and to a lesser extent Co2+ can replace iron in the activation process. The enzyme incorporates 4-5 mol iron/mol and 4.5-6 mol sulphide/mol during activation. In this process an [Fe-S] cluster appears to be built up, as indicated by optical and electron paramagnetic resonance (EPR) spectroscopy. In activated samples exposed to air the [Fe-S] cluster is EPR-detectable through an axial signal with g = 2.01 and g = 2.029 whose temperature and power saturation characteristics were similar to those of other [3Fe-xS] clusters. The activated enzyme, however, is readily inactivated even upon minor manipulation with destruction of the iron-sulfur core. PMID- 2990925 TI - Activation of protein kinase isozymes by cyclic nucleotide analogs used singly or in combination. Principles for optimizing the isozyme specificity of analog combinations. AB - 104 cAMP analogs, most of them modified in the adenine moiety, were tested as activators of cAMP-dependent protein kinase I (from rabbit or rat skeletal muscle) and kinase II (from bovine heart or rat skeletal muscle). When tested singly, only 2-phenyl-1,N6-etheno-cAMP showed a considerably (sevenfold) higher potency as an activator of kinase II than of kinase I. Analogs containing an 8 amino modification preferentially activated kinase I, some being more than 10 fold more potent as activators of kinase I than kinase II. When two analogs were combined, the concentration of one (complementary) analog required to half maximally activate each isozyme was determined in the presence of a fixed concentration of another (priming) analog. Analogs tested in combination had been analyzed for their affinity for the intrasubunit binding sites (A, B) of isozyme I and II. The degree to which complementary analogs preferentially activated one isozyme was plotted against the mean site selectivity, i.e. (affinity A/B isozyme I X affinity A/B isozyme II) 1/2. This plot produced a straight line, the slope of which reflected the ability of the priming analog to discriminate homologous sites on the isozymes. This means that the isozyme discriminating power of an analog pair can be quantitatively predicted from the affinity of the analogs for site A and B of the two enzymes. It also means that a systematic analysis of those features of analogs imparting a high mean site selectivity or the ability to discriminate between homologous isozyme sites will facilitate the synthesis of new even more isozyme-selective analogs. PMID- 2990926 TI - Nucleotide sequence of the Escherichia coli gap gene. Different evolutionary behavior of the NAD+-binding domain and of the catalytic domain of D glyceraldehyde-3-phosphate dehydrogenase. AB - A 1523-base-pair DNA fragment, spanning the gap gene from Escherichia coli, has been sequenced. It contains an open-reading frame whose length (330 amino acids) is in agreement with D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecular mass. This coding sequence is preceded by a Shine-Dalgarno complementary sequence and by two overlapping promoter-like structures. The codon usage within gap is consistent with that expected for a gene which is strongly expressed. The amino acid sequence of the E. coli GAPDH, deduced from the DNA sequence, contains all the amino acids postulated to play a functional role in GAPDH. Comparison of the E. coli enzyme with enzymes from other species reveals different evolutionary behaviour of the NAD+-binding domain and of the catalytic domain of GAPDH. The E. coli enzyme is found to be more similar to eucaryotic enzymes than to enzymes from thermophilic bacteria. This observation is discussed in terms of adaptation to growth at high temperature. PMID- 2990927 TI - Human red-blood-cell Ca2+-antagonist binding sites. Evidence for an unusual receptor coupled to the nucleoside transporter. AB - The human red blood cell ghost Ca2+-antagonist binding sites were characterized with (+/-)-[3H]nimodipine. The labelled 1,4-dihydropyridine bound in a non cooperative, reversible manner with a Kd of 52 nM at 25 degrees C to 9.65 pmol sites/mg ghost protein. The stereochemistry of the binding domain was evaluated with the optically pure enantiomers of chiral 1,4-dihydropyridines. In contrast to the 1,4-dihydropyridine-selective receptors on Ca2+ channels in electrically excitable tissues, the (+) enantiomer of nimodipine and the (-) enantiomer of the benzoxadiazol 1,4-dihydropyridine (PN 200-110) were bound with higher affinity than the respective optical antipodes. The human red blood cell ghost [3H]nimodipine-labelled sites also interacted with the inorganic Ca2+-antagonist La3+ (increase in the number of binding sites), and were allosterically regulated by the optical enantiomers of the phenylalkylamine-type Ca2+-antagonists (e.g. verapamil, desmethoxyverapamil, methoxyverapamil). The benzothiazepines d- or l cis-diltiazem were without effect. Nucleosides (adenosine approximately equal to inosine greater than cytidine) were inhibitory at the nimodipine-labelled site, as were the nucleoside uptake inhibitors dipyridamole, hexobendine, dilazep, nitrobenzylthioinosine and nitrobenzylthioguanosine. The binding sites have essential sulfhydryl groups, show trypsin sensitivity, but are relatively heat stable. When nitrobenzylthioinosine was employed as a covalent probe to inactivate the red blood cell ghost nucleoside carrier, [3H]nimodipine binding was irreversibly lost. (+)-Nimodipine greater than (-)-nimodipine inhibited [14C]adenosine transport into human red blood cells. A good correlation between IC50 values for inhibition of [3H]nimodipine binding and IC50 values for inhibition of [14C]adenosine uptake was found for 18 compounds. Sheep red blood cells (which lack the nucleoside transporter) had no detectable [3H]nimodipine binding sites. It is concluded that the Ca2+-antagonist receptor sites of the human erythrocyte are coupled to the nucleoside transporter. PMID- 2990929 TI - 16th Sir Hans Krebs lecture. The symbiosis of metal and protein functions. PMID- 2990928 TI - Diastereomers of adenosine 3',5'-monothionophosphate (cAMP[S]) antagonize the activation of cGMP-dependent protein kinase. AB - cGMP-dependent protein kinase contains four cGMP-binding sites which are homologous to the four cAMP-binding sites of cAMP-dependent protein kinase. The interaction of the diastereomers of adenosine 3',5'-thionophosphate, (PS)-cAMP[S] and (PR)-cAMP[S], with cGMP-dependent protein kinase has been studied. Autophosphorylation of cGMP-dependent protein kinase is stimulated by cAMP and (PS)-cAMP[S] with apparent KA values of 7 microM and 94 microM, respectively. cAMP-stimulated autophosphorylation is inhibited competitively by (PR)-cAMP[S] with a Ki value of 15 microM. The phosphorylation of the peptide substrate (Leu Arg-Arg-Ala-Ser-Leu-Gly) is stimulated by cGMP (approx. KA 1 microM) and cAMP (approx. KA 98 microM) but neither by the (PR) nor (PS) stereoisomer of cAMP[S]. (PR)-cAMP[S] and (PS)-cAMP[S] inhibit competitively cAMP-or cGMP-stimulated phosphorylation of the peptide substrate with Ki values of 52 microM and 73 microM, respectively. (PS)-cAMP[S] stimulates the phosphorylation of the peptide substrate by an autophosphorylated enzyme. Binding of [3H]cGMP to cGMP-dependent protein kinase is inhibited by (PS)-cAMP[S] and (PR)-cAMP[S] with IC50 values of 200 microM and 15 microM, respectively. These results show that both diastereomers of cAMP[S] bind to cGMP-dependent protein kinase. (PR)-cAMP[S] has properties of a pure antagonist whereas (PS)-cAMP[S] has properties of a partial agonist. The results provide further evidence that autophosphorylation of the enzyme affects the interaction between the cGMP-binding sites and the catalytic center of the enzyme by facilitating the activation of the phosphotransferase reaction. PMID- 2990930 TI - Characterization of the bovine lens plasma membrane substrates for cAMP-dependent protein kinase. AB - cAMP-dependent protein kinase, derived from either calf lens or bovine heart, promotes the phosphorylation of three lens plasma membrane proteins of molecular mass 28 kDa, 26 kDa and 18 kDa. Correlation of the maximal level of phosphorylation of these components with the Coomassie blue staining intensity of fractionated lens membranes suggests that the phosphorylation of the 28 kDa and 18 kDa components may be approximately stoichiometric. The protein kinase substrates could be dephosphorylated by a cardiac sarcoplasmic-reticulum-bound protein phosphatase activity. The 26 k Da component comigrated with MP26, the major lens membrane component that has been localized to the lens fiber cell junction. Treatment of phosphorylated lens membranes with chymotrypsin did not suggest that any of the three major phosphorylated components was derived from the partial proteolysis of a larger phosphoprotein. After electrophoretic separation of phosphorylated proteins, treatment with N-chlorosuccinimide confirmed that there was little similarity in the structure of the three phosphoproteins. Chymotrypsin did, however, reveal a cryptic phosphorylation site in a 22 kDa fragment that appeared to be derived from MP26. Treatment of phosphorylated membranes with reducing agents resulted in the disappearance of the 28 kDa phosphorylated component and the appearance of a new phosphorylated component of 18 kDa; neither MP26 nor the original 18 kDa component was affected by such treatment. It is not clear whether the original 18 kDa phosphoprotein, present in unreduced samples, is the same as that generated with reducing agents from the 28 kDa phosphorylated lens membrane component. PMID- 2990931 TI - Characterization of the binding sites for nimodipine and (-)-desmethoxyverapamil in bovine cardiac sarcolemma. AB - The bovine cardiac sarcolemmal binding sites for the dihydropyridine nimodipine and the phenylalkylamine (-)-desmethoxyverapamil were studied. The density of the nimodipine and (-)-desmethoxyverapamil binding sites increased 8.3-fold and 3.4 fold with the sarcolemma. The binding sites for both compounds were destroyed by trypsin. Nimodipine bound in the presence of 1 mM free calcium to a high-affinity and a low-affinity site with apparent Kd values of 0.35 +/- 0.09 nM (n = 9) and 33 +/- 6.0 nM (n = 9) and with apparent densities of 0.3 +/- 0.05 pmol/mg (n = 9) and 8.2 +/- 1.0 pmol/mg (n = 9). The binding to the high-affinity site was abolished by 1 mM EGTA. The binding sites were specific for dihydropyridines. The (-)-isomers of several phenylalkylamines inhibited nimodipine binding by an apparent allosteric mechanism. (-)-Desmethoxyverapamil bound in the presence of 5 mM EGTA to a high-affinity and a low-affinity site with apparent Kd values of 1.4 +/- 0.3 nM (n = 6) and 171 +/- 26 nM (n = 6) and with apparent densities of 0.16 +/- 0.02 pmol/mg (n = 6) and 13.6 +/- 2.7 pmol/mg (n = 6). The binding to both sites was inhibited by calcium with a half-maximal concentration of 4.3 mM. The binding sites were specific for the other phenylalkylamines and had a higher affinity for the (-)-isomers than for the (+)-isomers. Nimodipine inhibited the binding of (-)-desmethoxyverapamil by an apparent allosteric mechanism. d-cis Diltiazem inhibited non-competitively the binding of (-)-[3H]desmethoxyverapamil with a Ki of 3.7 microM. Diltiazem up to concentrations of 10 microM did not affect the amount of nimodipine bound at equilibrium at 20 degrees C. However, but in agreement with this result, diltiazem decreased threefold at 20 degrees C the dissociation and association rates for the high-affinity nimodipine receptor. These rates were only marginally affected at 4 degrees C and 37 degrees C. d-cis Diltiazem reversed in a competitive manner the inhibition of nimodipine binding elicited by the addition of (-)-desmethoxyverapamil with a Ka value of 1.6 microM. The amount of nimodipine bound was inhibited by 50% by the adenosine uptake inhibitors nitrobenzylthioinosine and hexobendine with apparent median inhibitory concentrations of 1 nM and 3 nM, respectively. Nitrobenzylthioinosine completely abolished binding of nimodipine to the low-affinity site, but did not affect binding to the high-affinity site.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2990932 TI - Cytochrome b, the var 1 protein, and subunits I and III of cytochrome c oxidase are synthesized without transient presequences in Saccharomyces cerevisiae. AB - The N-termini of four mitochondrial translation products, the var 1 protein, cytochrome b, and subunits I and III of cytochrome c oxidase have been characterized in Saccharomyces cerevisiae and compared with the known DNA sequences of the respective structural genes. The four mature proteins correspond to the predicted primary translation products and retain the formylated methionine residue. Thus, subunit II of cytochrome c oxidase studied previously [Pratje et al. (1983) EMBO J.2, 1049-1054] is so far the only mitochondrial translation product carrying a N-terminal-extended transient presequence in S. cerevisiae. PMID- 2990933 TI - Atypical phenylketonuria with "dihydrobiopterin synthetase" deficiency: absence of phosphate-eliminating enzyme activity demonstrated in liver. AB - An assay for the phosphate-eliminating enzyme (PEE) activity in liver was developed which required only 5-10 mg tissue. PEE catalyses the elimination of inorganic triphosphate from dihydroneopterin triphosphate, which is the second and irreversible step in the biosynthesis of tetrahydrobiopterin (BH4). In the presence of substrate, magnesium, NADPH, and a sepiapterin reductase fraction from human liver, PEE catalysed the formation of BH4 which was measured by HPLC and electrochemical detection. In adult human liver, a PEE activity of 1.02 +/- 0.134 microU/mg protein (mean +/- 1 SD; n = 5) was observed. In liver needle biopsy material from five patients with defective biopterin biosynthesis, no PEE activity was found (less than 2% and 6% of the control values, respectively). The presence of an endogenous inhibitor was excluded. In a patient who died without definite diagnosis and in a patient with beta-thalassaemia liver PEE activity was increased. Sepiapterin reductase activity was present in all cases. Results indicate that in "dihydrobiopterin synthetase" deficiency, the most frequent of the rare BH4-deficient variants of hyperphenylalaninaemia, the molecular defect consists in a defect of PEE. PMID- 2990935 TI - Antigens and circulating immune complexes related to the primate retroviral glycoprotein SiSV gp70: prevalence and distribution in human sera. AB - We have shown previously that antigens and also circulating immune complexes related to the primate retroviral envelope glycoprotein SiSV gp70 correlate with early mortality and survival of 56 patients with acute leukemias and chronic myelogenous leukemia in blast crisis. The prevalence and general distribution of these antigens and immune complexes in human sera was therefore of obvious interest. We now report an analysis of a total of 200 sera from 166 patients. Of these 113 sera were obtained from 84 patients with acute or chronic leukemias and 87 from 82 non-leukemic subjects, including laboratory workers and patients with non-leukemic neoplasias. Antigens and immune complexes were determined by enzyme linked immunosorbent assays (ELISA). The anti-SiSV gp70 antiserum used predominantly recognized the protein moieties of the glycoproteins. The distribution of SiSV gp70-related antigens and immune complexes was similar among leukemic and non-leukemic sera. The prevalence of SiSV gp70-related antigens was 53% and of SiSV gp70-related immune complexes 49% in all sera. SiSV gp70-related antigens were detected in a somewhat higher proportion of non-leukemic (69%) than leukemic sera (40%), whereas SiSV gp70-related immune complexes and cross reactive antibodies were more evenly distributed in leukemic and non-leukemic sera (in 46 and 51% of leukemic and 54 and 51% of non-leukemic sera). Presence of antigens correlated with presence of SiSV gp70-related immune complexes in 71% of all sera, but in 13% of all sera antigens were detectable only by determining SiSV gp70-related immune complexes. Total circulating immune complexes did not correlate with SiSV gp70-related immune complexes. The origin and pathophysiological role of the antigens are discussed. PMID- 2990936 TI - Effect of in vitro non-enzymatic glycosylation of human skin collagen on susceptibility to collagenase digestion. AB - The effect of glycosylation on susceptibility of skin collagen to collagenase digestion was studied in a skin sample obtained at autopsy from the interscapular region of a 24 year old white male who had died of an acute illness and who had no history of diabetes. Homogeneous suspensions of insoluble collagen were prepared, and were incubated in 50 mmol l-1 dextrose at pH 7.35 and 37 degrees C for 7 days. Non-enzymatic glycosylation measured by the weak acid hydrolysis/thiobarbituric acid method increased from 13.1 +/- 1.0 (n = 5) to 45.2 +/- 5.5 (n = 8) nmol fructose per 10 mg collagen (P less than 0.001). Digestion of collagen using clostridial collagenase was monitored by measuring (a) hydroxyproline content and (b) absorption at 206 nm of the supernatant after centrifugation to remove substrate. The rate of digestion was similar in glycosylated and control collagen. We conclude that the ketoamine link formed in non-enzymatic glycosylation does not increase the resistance of collagen to enzymatic digestion. The possibility remains that subsequent rearrangement of this link could be important in this respect. PMID- 2990934 TI - Moyamoya disease with a developmental anomaly of the mesenchyme. AB - We treated a Japanese boy with moyamoya disease accompanied by synbrachydactylia, funnel chest, pes equinus and short stature. Angiograms showed the anomalous origin of the occipital artery in addition to the moyamoya vascular network at the base of the brain. A generalised mesenchymal anomaly was suggested in this case. PMID- 2990937 TI - [125I]N6-(3-iodo-4-hydroxyphenyl)isopropyladenosine: the use of the diastereomers as ligands for adenosine receptors in rat brain. AB - The diastereomers of N6-(4-hydroxyphenyl)isopropyladenosine (HPIA) were synthesized, radioiodinated and used as ligands for adenosine receptors in rat brain membranes. In contrast to the S-isomer, specific binding (measured by displacement with 10 microM N6-R-phenylisopropyladenosine (PIA] of [125I]N6-[R-(3 iodo-4-hydroxyphenyl)isopropyl] adenosine ([125I]R- IHPIA) required about 2 h for equilibrium but bound with a higher affinity. Scatchard analysis of binding data were compatible with the existence of single binding site with Kd values of 0.7 nM (R-isomer) and 10 nM (S-isomer), and maximal binding of 228 and 237 fmol/mg, respectively, at 30 degrees C. Computer-based non-linear curve fitting resulted in estimates of 0.67 and 7.5 nM, and of 208 and 173 fmol/mg, for the R- and S isomer, respectively. Dissociation of the R-isomer in the presence of 10 microM PIA was biphasic, both phases being increased in the presence of 100 microM Gpp(NH)p. The ratio of the rate of dissociation for the slower phase, k2, and the rate of association, k1, yielded a further estimate for Kd of 0.3 nM. The specific binding of [125I]R-IHPIA was displaced by adenosine receptor agonists with the following order of decreasing potency: N6-[R-(phenyl) isopropyl]adenosine = N6-[R-(4-hydroxyphenyl)isopropyl]adenosine greater than 5' N-ethylcarboxamidoadenosine greater than 2-chloroadenosine greater than N6-[S (phenyl)isopropyl]adenosine = N6-[S-(4-hydroxyphenyl)isopropyl]adenosine which is typical of binding to Ri/A1 type receptors. Receptor antagonists displaced this binding in the following order of decreasing potency: 8-(p-hydroxyphenyl) theophylline greater than 8-phenyltheophylline greater than theophylline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990939 TI - Role of endothelium in the binding of [3H]prazosin to the aortic alpha 1 adrenergic receptors. PMID- 2990938 TI - Behavioural sensitivity to PIA in selectively bred mice is related to a number of A1 adenosine receptors but not to cyclic AMP accumulation in brain slices. AB - In a dose of 0.1 mg/kg, PIA had marked behavioural effects in long-sleep mice (which show a high sensitivity to ethanol, while no significant effect was observed in short sleep mice (low sensitivity to ethanol). The number of [3H]PIA binding sites in cortex and subcortical brain regions was significantly higher in long-sleep than in short-sleep mice. The KD value was higher in cortex and cerebellum in the short-sleep mice, but there were no differences in the number of hippocampal beta-adrenoceptors or in the adenosine analogue-induced increase in cyclic AMP accumulation in slices of mouse hippocampus. PMID- 2990940 TI - Rat atrial natriuretic peptide elevates the level of cyclic GMP in astroglia-rich brain cell cultures. PMID- 2990942 TI - Trimipramine, a tricyclic antidepressant exerting atypical actions on the central noradrenergic system. AB - Trimipramine is a tricyclic antidepressant which has only weak effects on noradrenergic systems. Its mechanism of action is not understood, but its clinical effectiveness has been proven over a period of 20 years. In the present investigation, trimipramine was shown to have no effect on noradrenaline (NA) stimulated adenylate cyclase activity after either acute or once daily application for 28 days. There was no change in the KD or Bmax of [3H]DHA binding, demonstrating a lack of effect on beta-adrenergic receptors. However, it did exert effects on the cortical noradrenergic projection of the locus coeruleus and these effects were opposite to those described for typical antidepressants. In acute experiments, systemic injections of trimipramine potently activated locus coeruleus neurons and produced a reduction in the depressant action of noradrenaline administered iontophoretically to neurons in the cingulate cortex. After 4 weeks' treatment with a daily injection of 30 mg/kg i.p. trimipramine, cingulate cortical neurons became supersensitive to the action of iontophoretically applied noradrenaline. Although it is unknown whether these effects are related to the therapeutic efficacy of trimipramine, the results demonstrate that the down-regulation of central noradrenergic sensitivity in the rat is not a prerequisite for clinical efficacy of antidepressants in man. PMID- 2990941 TI - Pulmonary macrophages induce deterioration of guinea-pig tracheal beta-adrenergic function through release of oxygen radicals. PMID- 2990943 TI - Inhibition by yohimbine of the calcium-dependent evoked release of [3H]GABA in rat and mouse brain slices in vitro. AB - Calcium-dependent release of [3H]GABA was elicited by electrical stimulation in slices of rat and mouse cerebral cortex or by potassium stimulation in the mouse brain-stem. The stimulation-evoked release of [3H]GABA was inhibited by yohimbine in a concentration-dependent manner. High concentrations of other alpha adrenoceptor antagonists such as phentolamine, RS 21361, idazoxan, rauwolscine and corynanthine also inhibited [3H]GABA release. This effect was not observed with pseudoyohimbine or prazosin. [3H]GABA release was not affected by exposure to the alpha-adrenoceptor agonists clonidine, M7, noradrenaline or methoxamine. In addition, clonidine did not antagonize the yohimbine-induced inhibition of [3H]GABA release. The inhibitory effect of yohimbine did not result from an interaction with endogenously released noradrenaline since the inhibition was still observed in reserpine-pretreated animals. It is concluded that yohimbine and other alpha 2-adrenoceptor antagonists inhibit the stimulation-evoked release of [3H]GABA through a mechanism which appears to be independent of the blockade of alpha 2-adrenoceptors and which does not involve an interaction with endogenous noradrenaline. The present results indicate that yohimbine exerts non specific actions on the release of [3H]GABA and that similar effects can be observed with other alpha-adrenoceptor blocking agents in high concentrations. Consequently, when studying the effects of yohimbine and other alpha 2 adrenoceptor antagonists on noradrenergic neurotransmission, the possibility of non-specific effects should be taken into consideration, particularly in the high concentrations range. PMID- 2990944 TI - Evidence for two types of P2-purinoceptor in longitudinal muscle of the rabbit portal vein. AB - The actions of ATP, 2-methylthioATP, alpha, beta-methyleneATP, beta, gamma methyleneATP, adenosine and non-adrenergic, non-cholinergic inhibitory nerve stimulation were compared on the ergotamine-contracted longitudinal muscle of the isolated rabbit portal vein with the vessel endothelium either intact or removed by mechanical rubbing. Non-adrenergic, non-cholinergic inhibitory nerve stimulation produced frequency-dependent relaxations. The purines (except alpha, beta-methyleneATP) also caused relaxation with the following potency order: 2 methylthioATP greater than ATP greater than beta, gamma-methyleneATP = adenosine. 8-Phenyltheophylline, a potent P1-purinoceptor antagonist, antagonised relaxations to adenosine but not those to ATP indicating that ATP and its analogues act directly via a P2-purinoceptor and not via a P1-purinoceptor after breakdown to adenosine. alpha, beta-MethyleneATP produced contractions which were not maintained and was tachyphylactic. The contractions were unaffected by tetrodotoxin and therefore unlikely to be due to release of a contractile substance from a neuronal source following initiation of an action potential. Removal of the endothelium did not affect the actions of non-adrenergic, non cholinergic inhibitory nerve stimulation, ATP, alpha, beta-methyleneATP, beta, gamma-methyleneATP or adenosine. Thus purine action does not have an endothelium dependent component in the rabbit portal vein longitudinal muscle. The results indicate the possibility that there may be more than one type of P2-purinoceptor in this tissue. PMID- 2990945 TI - Effect of [D-Ala2,Met5]enkephalinamide and [D-Ala2,D-Leu5]enkephalin on cholinergic and noradrenergic neurotransmission in isolated atria. AB - In rabbit isolated atria, [D-Ala2,Met5]enkephalinamide and [D-Ala2,D Leu5]enkephalin (0.1-3 microM) inhibited responses to cholinergic nerve stimulation in a concentration-dependent manner without affecting responses to exogenous acetylcholine. The inhibitory effect was blocked by the opiate receptor antagonist naloxone (1 microM). In rabbit atria in which the transmitter acetylcholine stores had been radioactively labelled by preincubating the tissue in [3H]choline, tetrodotoxin (100 ng/ml) significantly (P less than 0.001) blocked the stimulation-induced (2 Hz for 3 min) release of radioactivity. Both [D-Ala2,Met5]enkephalinamide and [D-Ala2,D-Leu5]enkephalin (0.3 and 1 microM) significantly decreased stimulation-induced radioactivity release and their effects were blocked by naloxone (1 microM). In rat isolated atria, [D Ala2,Met5]enkephalinamide and [D-Ala2,D-Leu5]enkephalin (0.3-3 microM) inhibited responses to cholinergic nerve stimulation without affecting responses to exogenous acetylcholine. The inhibitory effect was blocked by naloxone (1 microM). In guinea-pig isolated atria, responses to cholinergic nerve stimulation were unaffected by the enkephalin analogues. In rabbit, rat and guinea-pig isolated atria, responses to noradrenergic nerve stimulation and exogenous noradrenaline were unaffected by the enkephalin analogues. PMID- 2990946 TI - Actions of betahistine at histamine receptors in the brain. AB - The actions of betahistine (N alpha-methyl-2-pyridylethylamine) on brain histamine receptors were investigated in a series of biological models. [3H]Mepyramine binding to H1-receptors in membranes from guinea-pig cerebellum was inhibited by betahistine with a Ki value of 31 microM. The binding of [3H]mepyramine in brain of the living mouse was inhibited by betahistine in high dosages (150-300 mg/kg). In slices from mouse cerebral cortex, betahistine induced [3H]glycogen hydrolysis in a concentration-dependent manner with an EC50 value of 9.0 microM with a maximal effect 57% that of histamine. Mepyramine and triprolidine, two H1-receptor antagonists, inhibited the betahistine-induced glycogenolysis with Ki values of 28 nM and 7 nM respectively. In slices from guinea-pig hippocampus, betahistine stimulated the accumulation of cyclic AMP in the presence of 5 microM impromidine, a H2-receptor agonist. The maximal effect represented 22% of that elicited by histamine at the H1-receptor and the EC50 value was 32.4 microM. Mepyramine at 0.1 microM partially blocked the response to betahistine. Together these various observations indicate that betahistine is a partial agonist at cerebral H1-receptors. Finally, betahistine was not an agonist at histamine H3-autoreceptors but was a rather potent antagonist of the inhibitory effect of exogenous histamine on [3H]histamine release elicited by K+ depolarisation in slices from rat cerebral cortex (Ki = 6.9 microM). PMID- 2990947 TI - Dissociation of cysteine and glutathione levels from nitroglycerin-induced relaxation. AB - The present study investigated possible involvement of cysteine (CSH) and reduced glutathione (GSH) as critical cellular sulfhydryls which mediate nitroglycerin (GTN)-induced cyclic GMP accumulation and relaxation in bovine coronary artery (BCA). Tolerance to the relaxant effects of GTN was induced in BCA in vitro by preincubation with 1 mM GTN for 2 h. GTN-tolerant BCA were at least 100-fold less sensitive than non-tolerant BCA to the relaxant effects of GTN. Consistent with a relationship between tolerance to both GTN-induced cyclic GMP accumulation and relaxation, cyclic GMP accumulation induced by 1 microM GTN was markedly reduced in GTN-tolerant BCA when compared with non-tolerant BCA. Incubation with 1 mM CSH for 1 h did not significantly alter GTN-induced cyclic GMP accumulation or relaxation in either GTN-tolerant or non-tolerant BCA. Levels of CSH, GSH and glutathione-disulfide (GSSG) were measured in non-tolerant BCA, GTN-tolerant BCA and GTN-tolerant BCA incubated with 1 mM CSH for 1 h. Levels of CSH and GSH were lower in GTN-tolerant BCA than in non-tolerant BCA, whereas GSSG levels were similar in both. In GTN-tolerant BCA incubated with 1 mM CSH, CSH levels were more than 10-fold above, and GSH levels were similar to corresponding values obtained in non-tolerant BCA. These data indicate that although incubation with CSH did not significantly reverse tolerance to GTN-induced cyclic GMP accumulation and relaxation in BCA, it did effectively raise the level of CSH and GSH in GTN-tolerant BCA, at least to corresponding levels found in non-tolerant BCA. These results indicate that the relaxant effects of GTN in BCA do not correlate with tissue levels of CSH and GSH. The findings do not support the hypothesis that CSH and GSH are the cellular sulfhydryls involved in mediating GTN-induced guanylate cyclase activation, cyclic GMP accumulation and relaxation in intact BCA. PMID- 2990948 TI - Effects of K+ on the interaction between cardiac glycosides and Na,K-ATPase. AB - Inhibition of Na,K-ATPase by cardiac glycosides is at least partially antagonized by K+. The kinetics of the antagonism, however, appear complicated because K+ is capable of reducing both association and dissociation rate constants for the glycoside-enzyme interaction. In order to better understand the effect of K+, inhibition of partially purified Na,K-ATPase obtained from rat brain, guinea-pig heart and rat heart by ouabain, digoxin, digoxigenin, dihydrodigoxin and cassaine were compared in the presence of 1, 3 or 10 mM K+. Higher concentrations of K+ caused a parallel shift to the right in the concentration-inhibition curves for these compounds. For ouabain or digoxin, the extent of the shift was minimal with rat brain enzyme, intermediate with guinea-pig heart enzyme and more substantial with rat heart enzyme. For digoxigenin, dihydrodigoxin or cassaine, the extent of the shift was substantial in all enzyme preparations. These results could not be explained from either the affinity of the enzyme for the compound or its lipid solubility alone. The concentrations of these compounds required to cause a 50 percent inhibition of enzyme activity were markedly different with rat brain enzyme, but relatively similar with rat heart enzyme. The effects of K+, which depend on the source of the enzyme and chemical structures of the compounds, have to be considered in studies on comparative effects of various compounds on Na,K ATPase, [3H]ouabain binding, sodium pumping and the force of myocardial contraction. PMID- 2990949 TI - Cross-tolerance to the hypothermic effect of delta 8-tetrahydrocannabinol 11 hydroxy-delta 8-tetrahydrocannabinol and chlorpromazine in the mouse. AB - When a 5 mg/kg i.v. dose of delta 8-tetrahydrocannabinol (delta 8-THC) and 11 hydroxy-delta 8-tetrahydrocannabinol (11-OH-delta 8-THC) was administered daily to mice, tolerance to their hypothermic effects developed quickly. Cross tolerance to the hypothermic effect also developed between delta 8-THC and 11-OH delta 8-THC. Tolerance development was dose-dependent and dose-effect experiments indicated that the magnitude of the tolerance developed to 11-OH-delta 8-THC was greater than that to delta 8-THC. Both cannabinoids were cross-tolerant to chlorpromazine in the hypothermic effect, but not to morphine, pentobarbital and reserpine under the conditions used. The present results suggest that 11-OH-delta 8-THC has some role in the development of tolerance to the hypothermic effect of delta 8-THC in mice. PMID- 2990950 TI - The proenkephalin A fragment, peptide E: central processing and CNS activity in vivo. AB - The proenkephalin A derivative, peptide E, delayed gastrointestinal transit in mice and inhibited the micturition reflex in anesthetized rats after intracerebroventricular (i.c.v.) administration. BAM22P, BAM12P and [Met5]enkephalin, possible processing fragments of peptide E, were also compared in the two test systems. Of these peptides, peptide E and BAM 22P were found to have the greatest potency and activity. Studies in vitro of peptide E metabolism by enzyme homogenates of mouse brain using HPLC techniques revealed that peptide E is bound to the membrane homogenate avidly for an extended period of time. The total formation of BAM22P, BAM12P, [Met5]enkephalin and all other peptide fragments during a 40 min incubation period accounted for only 8% of the total peptide E added to the homogenates. Thus, peptide E, rather than one of its known metabolites, appears to be of primary importance in the initiation of CNS mediated effects. Further, these effects are probably the result of mu-opioid receptor activation. PMID- 2990951 TI - Differential effects of dextrorphan and levorphanol on the excitation of rat spinal neurons by amino acids. AB - The effects of the stereoisomers dextrorphan and levorphanol on the excitation of spinal neurons by electrophoretically administered excitatory amino acids were studied in pentobarbitone-anaesthetised rats. Both isomers reduced responses to N methyl-DL-aspartate (NMA), dextrorphan being both more selective and more potent than levorphanol in this respect. This observation supports the proposal that the NMA-blocking activity of a variety of drugs with psychotomimetic properties is subserved by actions at phencyclidine (PCP)/sigma opiate receptors. PMID- 2990952 TI - Effects of aniline on neuromuscular transmission. AB - Aniline hydrochloride increased the amplitude of the end-plate potentials of the frog sartorius muscle. Statistical analysis showed that the effects of aniline were purely presynaptic. The shape of the dose-release curve was similar to that of the 4-aminopyridine curve although the potency was weaker than that of 4 aminopyridine. The molecular configuration and the effect of aniline suggested that the site of action of aniline was the same as that of 4-aminopyridine. PMID- 2990953 TI - Increased affinity and selectivity of enkephalin tripeptide (Tyr-D-Ala-Gly) dimers. AB - The binding of alkylendiamide dimers of the three N-terminal residues of [D Ala2,D-Leu5]enkephalin (DADL) to rat brain and Ng108-15 neuroblastoma-glioma cell membranes was compared with that of DADL, Tyr-D-Ala-Gly-NMe-Phe-Gly-ol (DAGO) and morphiceptin. Tritiated DADL and DAGO were used as labeled ligands for delta- and mu-receptors, respectively. Dimerization of the tripeptides resulted in dramatic increases in both mu and delta binding. The binding to mu-receptors showed two peaks at an alkyl chain length of n = 2 and approximately n = 16. In contrast, delta binding (NG108-15 cells) increased steadily with increasing chain length. The dimers with n less than 18 were mu-preferential, and the one with n = 2 showed the most dramatic increase in mu selectivity with a 400 fold higher affinity to mu- than to delta-receptors. For long-chain alkyl spacers the compounds became delta selective. PMID- 2990954 TI - Anticonvulsant action of stereoisomers of gamma-glutamylaminomethylsulphonic acid in mice. AB - The effects of stereoisomers of gamma-glutamylaminomethylsulphonic acid (GAMS) on convulsions produced by intracerebroventricular injections of excitatory amino acids were studied in mice. gamma-D-GAMS preferentially blocked myoclonic seizures induced by kainate and had less pronounced anticonvulsant effect against quisqualate, N-methyl-D-aspartate, quinolinate, D-homocysteine sulphinate and L glutamate. gamma-L-GAMS displayed only a relatively weak protective effect against kainate- and, to some extent, quisqualate-induced seizures, with little or no effect on the convulsant properties of the other excitatory amino acids. These results indicate that the D configuration of the gamma-carbon atom of excitatory amino acid antagonists is preferred not only for NMDA receptor antagonism but for antagonism at non-NMDA receptors as well. PMID- 2990955 TI - Autoradiographic localization of delta opioid receptors in the rat brain using a highly selective bis-penicillamine cyclic enkephalin analog. PMID- 2990956 TI - The importance of the alpha-adrenoceptor agonist activity of dobutamine to inotropic selectivity in the anaesthetized cat. AB - Doses of dobutamine (steady-state infusions) required to increase inotropy (as measured by isovolumic indices of contractility) were lower than those required to produce tachycardia in the anaesthetized cat. When compared to isoprenaline, dobutamine produced less tachycardia for common increases in inotropy and thus demonstrated inotropic selectivity in this model. Dobutamine also produced mild pressor effects which were potentiated by beta-adrenoceptor blockade with propranolol. In efforts to define the role of the partial agonist activity of dobutamine for alpha-adrenoceptors in the production of selective inotropy the effects of dobutamine infusions were observed in cats pretreated with the alpha adrenoceptor antagonist phentolamine. In phentolamine-treated cats dobutamine did not demonstrate inotropic selectivity and showed a relationship between increased inotropy and tachycardia which was not significantly different from that obtained with isoprenaline. In contrast, phentolamine did not change the relationship between isoprenaline-induced tachycardia and increased inotropy. These data suggest that the agonist activity of dobutamine for alpha-adrenoceptors could be responsible for selective inotropy in the anaesthetized cat probably by baroreceptor-mediated reflex modulation of heart rate and/or possible stimulation of inotropic cardiac alpha-adrenoceptors. PMID- 2990957 TI - Electrophysiological and receptor studies in rat brain: effects of clorgyline. AB - Acute high doses of clorgyline produce a rapid inhibition of monoamine oxidase type A (MAO A) in the rat brain, together with an increase in norepinephrine and a decrease in the firing rate of locus coeruleus (LC) neurones: this decrease is reversed by piperoxane, an alpha 2 antagonist. In control animals, piperoxane increases LC neuronal firing showing that these noradrenergic neurones are under alpha 2-adrenoceptor-mediated tonic inhibition. Chronic administration of clorgyline, like acute doses of this MAO A inhibitor, significantly decreases cell firing in the LC and the effect is partially reversed by piperoxane. Chronic clorgyline treatment also produces significant decreases in [3H]clonidine and [3H]dihydroalprenolol binding in cerebral cortex, receptor changes which are slightly greater in animals showing greater inhibition of LC neuronal firing: such receptor changes do not occur following a single exposure to clorgyline. Electrophysiological studies in hippocampal pyramidal cells show that the chronic clorgyline treatment does not significantly induced subsensitivity to NE in these adrenoreceptive cells. PMID- 2990958 TI - The effect of acute ethanol administration on GABA receptor binding in cerebellum and hypothalamus. AB - The effect of the intraperitoneal administration of ethanol on [3H]GABA binding and glutamic acid decarboxylase (GAD) in cerebellum and hypothalamus was investigated. Acute ethanol administration produced an increase in the binding capacity of the high affinity GABA binding sites and a decrease in the binding capacity of low affinity sites. A decrease in the binding capacity of the high affinity GABA binding sites and an increase in the binding capacity of the low affinity sites were observed in the hypothalamus. No apparent changes were detected in the binding affinities for the two types of GABA receptor sites in both brain areas following ethanol treatment. Ethanol enhanced GAD activity in the cerebellum and reduced GAD activity in the hypothalamus. Changes in GABA binding may be involved in some of the neuropharmacological effects of ethanol. PMID- 2990960 TI - The central vs. peripheral effects of clonidine on ACTH, corticosterone and glucose release. AB - The role of central vs. peripheral actions of clonidine was investigated in the rat following the separate and combined administration of clonidine and 2-deoxy-D glucose (2-DG). Clonidine or 2-DG alone stimulated serum glucose and corticosterone but hypothalamic noradrenaline neuronal activity and ACTH release were stimulated by 2-DG only. The stimulation of noradrenaline neuronal activity and ACTH by 2-DG were totally blocked by clonidine. It was concluded that the increases in serum glucose and corticosterone after clonidine administration were not mediated by central effects but that alpha 2-adrenoceptor agonism by clonidine was responsible for inhibition of hypothalamic noradrenaline neuronal activity and pituitary ACTH release. PMID- 2990959 TI - Effect of a novel tachykinin, substance K, on salivation in rats. AB - A study of the effects of substance K (SK) and its synthetic fragments on salivation was performed. The rank order of potency of tachykinins that elicited salivation was as follows: physalaemin greater than substance P greater than eledoisin greater than kassinin greater than Arg-SK-(1-10) greater than SK-(1-10) greater than SK-(3-10). SK-(6-10) showed no activity. Atropine (1 mg/kg i.v.) had no effect on SK-induced salivation. PMID- 2990961 TI - A specific alkylating ligand for phencyclidine (PCP) receptors antagonizes PCP behavioral effects. PMID- 2990962 TI - Visualization of calcitonin gene-related peptide receptors in the rat brain. PMID- 2990963 TI - Separate site(s) of action of optical isomers of 1-methyl-5-phenyl-5 propylbarbituric acid with opposite pharmacological activities at the GABA receptor complex. AB - The behavioral profile of the optical isomers of 1-methyl-5-phenyl-5 propylbarbituric acid (MPPB) and their interaction with the convulsant binding site at the GABA receptor complex were investigated. R(-)-MPPB produced dose related loss of righting reflex, whereas S(+)MPPB produced convulsions in a dose dependent manner. Subconvulsive doses of S(+)MPPB were proconvulsant with a subeffective dose of picrotoxin. S(+)MPPB-induced seizures were blocked by R( )MPPB and pentobarbital. In contrast, S(+)MPPB did not block the loss of righting reflex produced by R(-)MPPB or pentobarbital. S(+)MPPB inhibited the binding of [35S]t-butylbicyclophosphorothionate (TBPS), a ligand that binds to the picrotoxin site on the oligomeric GABA receptor complex, to rat brain membranes competitively, whereas R(-)MPPB inhibited it noncompetitively. Thus, the optical isomer of MPPB, which have opposite pharmacological effects, interact differently with the convulsant (TBPS) site at the GABA receptor complex. These results suggest that the convulsant S(+)MPPB and the depressant R(-)MPPB may produce their behavioral effects by acting via convulsant and anticonvulsant sites, respectively, at the GABA receptor complex. PMID- 2990964 TI - Chronic clonidine induces functional down-regulation of presynaptic alpha 2 adrenoceptors regulating [3H]noradrenaline and [3H]5-hydroxytryptamine release in the rat brain. AB - Clonidine inhibited, through the activation of alpha 2 presynaptic receptors, the release of [3H]noradrenaline (pIC30 = 7.47) and [3H]5-hydroxytryptamine (pIC30 = 6.47) evoked by 15 mM KCl from superfused rat cerebral cortex synaptosomes. The natural agonist noradrenaline inhibited the K+-evoked release of the two [3H]amines less effectively after long-term (12 days) treatment with clonidine than after acute treatment. It can be concluded that chronic clonidine administration can induce down-regulation of both the alpha 2 presynaptic autoreceptors located on noradrenaline terminals and the alpha 2 presynaptic heteroreceptors located on serotonin terminals. PMID- 2990965 TI - The kappa opioid receptor and food intake. AB - Many studies have suggested a role of opioid receptors in the modulation of food intake. Several distinct classes of opioid receptors have been postulated. In an attempt to establish which opioid receptor(s) modulate feeding we studied the effect of the kappa agonist, bremazocine, on feeding and compared its effects to the preferential mu agonist, morphine, and the mixed kappa-sigma agonist, butorphanol and the kappa agonist, ethylketocyclazocine. Bremazocine increased feeding to the same extent as morphine and was less potent than the mixed agonist/antagonists. The bremazocine effect demonstrated a bell-shaped dose response curve. Daily administration of bremazocine or morphine enhances the effect on increasing food intake. However, this effect of daily injections on enhancing food intake is not present when animals receiving morphine are crossed over to bremazocine and vice versa. The bremazocine effect is enhanced by diprenorphine and not inhibited by naloxone. Low doses of the dopamine antagonist, haloperidol, enhance the bremazocine effect and higher doses inhibit it. Finally, using another kappa agonist, tifluadom, we showed that the effect on food intake is stereospecific. Our studies provided further evidence for a role for the kappa opioid receptor in feeding. However, they also suggest that more than one subpopulation of opioid receptors is involved in feeding modulation. PMID- 2990966 TI - Species differences in the concentrations and distributions of opioid binding sites. AB - Binding at the mu, delta- and kappa-types of opioid binding sites was compared in homogenates from the brains of guinea-pig, rabbit, rat and two mouse strains, under conditions of selective labelling. Species differences were shown by two observations. Firstly, analysis of saturation curves in homogenates of brain from which the cerebellum had been removed showed that in guinea-pig brain the opioid binding sites consist of 24% mu-sites, 32% delta-sites and 44% kappa-sites. In contrast, in rabbit brain the corresponding values are 43% mu-sites, 19% delta sites and 37% kappa-sites and in rat brain, 46% mu-sites, 42% delta-sites and 12% kappa-sites. In the brains of DBA/2 mice the opioid binding sites are comprised of 51% mu-sites, 29% delta-sites and 20% kappa-sites and in C57BL/10 mice, of 44% mu-sites, 35% delta-sites and 21% kappa-sites; these strain differences are due to significant differences in the concentrations of the mu-sites. Secondly, species differences were found when the binding of single concentrations of tritiated ligands (1 X KD value in whole brain) was determined at mu-, delta- and kappa-sites in six brain regions from guinea-pig, rat or rabbit. PMID- 2990967 TI - Potentiating effects of 4-aminopyridine on responses to intramural vagal stimulation in the isolated dog atrium. AB - The effects of 4-aminopyridine (4-AP) on the chronotropic and inotropic responses to intramural parasympathetic nerve stimulation and to injection of acetylcholine (ACh) into the sinus node artery were investigated after treatment with propranolol in the isolated, blood-perfused dog atrium. 4-AP (3-100 micrograms) induced positive chronotropic and inotropic effects and 300 micrograms of 4-AP induced biphasic, negative and positive effects. The negative responses to 4-AP were completely blocked by 10 micrograms of atropine. 4-AP (more than 30 micrograms) potentiated significantly the negative chronotropic and inotropic responses to intramural parasympathetic nerve stimulation at a frequency of 3-30 Hz. The potentiation of the responses to stimulation was greater at a low than at a high frequency of stimulation. Potentiation of the chronotropic effect was greater than that of the inotropic one. 4-AP in the doses used did not change the negative chronotropic and inotropic responses to injection of ACh into the sinus node artery. These results suggest that the potentiation by 4-AP of the cardiac responses to parasympathetic nerve stimulation was due to an increase in release of ACh from nerve terminals but not to an increase in responsiveness at the effector site. PMID- 2990968 TI - Sabra rats as a model to differentiate between Na+ and GTP regulation of alpha 2 adrenoceptor densities. AB - Sodium ions and guanyl nucleotides play an important role in increasing alpha 2 adrenoceptor densities in cerebral and renal cortex of normotensive rats. The in vitro effect of Na+ and GTP was investigated on cerebral and renal alpha adrenoceptors in hypertensive (SBH, salt-sensitive) and normotensive (SBN, salt resistant) Sabra rats. In SBH and SBN rats, guanyl nucleotides increased cerebral and renal high-affinity alpha 2-adrenoceptor densities. Sodium ions, in contrast, markedly increased cerebral and renal high affinity alpha 2-adrenoceptor densities only in SBH rats. Under these conditions, alpha 1-adrenoceptor densities were unchanged. Thus, although Na+ and GTP both increase alpha 2 adrenoceptor densities, these agents appear to mediate their regulatory effects via different membrane components. Moreover, the absence of sodium regulation of alpha 2-adrenoceptors in SBN rats may be responsible for the resistance to salt induced hypertension. PMID- 2990969 TI - Effect of neuraminidase treatment on the inotropic response to ouabain, isoproterenol and calcium in the guinea pig heart. AB - To determine the role of the glycocalyx sialic acids residues in excitation contraction coupling and the inotropic response to cardiotonic agents, we studied the effect of neuraminidase treatment on the response to ouabain, isoproterenol, calcium and reduced extracellular sodium in Langendorff preparations of adult guinea pig hearts. Neuraminidase treatment (0.01 unit/ml, 1 h) reduced the magnitude of the positive inotropic response to 2.5 X 10(-7) M ouabain and the maximum response to 5 X 10(-7) M ouabain by about 46% and 30%, respectively, but did not prevent ouabain toxicity. Neuraminidase treatment did not affect the contractility produced by calcium concentration alterations up to 5 mM calcium or the positive inotropic effect produced by lowering external sodium to as low as 80 mM. The inotropic response to as high as 10(-8) M isoproterenol was also not affected. The contractility response developed to calcium concentrations greater than 5 mM and to 5 X 10(-8) M isoproterenol were significantly reduced (P less than 0.05) by neuraminidase treatment. The content of sialic acids in neuraminidase-treated hearts used in the above concentration-response studies of ouabain, isoproterenol, calcium, and sodium was reduced by 70.7%, 66.1%, 65.6% and 66.2%, respectively. Neuraminidase treatment had no effect on basal (Na+ - K+)ATPase and Mg2+ - ATPase activities of (Na+ - K+)ATPase-containing membrane preparations of the guinea pig left ventricle. Neuraminidase treatment neither influenced the sensitivity of the enzyme (Na+ - K+)ATPase to ouabain inhibition nor did it affect the characteristics of [3H]ouabain binding to the preparation. These results suggest that the sialic acids of the glycocalyx in the guinea pig left ventricle play an important role in part of the inotropic response to subtoxic concentrations of ouabain. PMID- 2990970 TI - Permissive role of spinal alpha 1-adrenoceptors in sudomotor efferents. AB - The electrodermal potential (EDP) recorded in the forepaws of anaesthetized cats in response to stimulation of the cholinergic-sympathetic nervous system at different levels was taken as a measure for sudomotor activity. Electrical stimulation of the hypothalamus with square wave pulses (1 ms duration, 0.5-64 Hz, 2 s train length) at intervals of 1 min induced rate-dependent EDPs which were inhibited at all rates of stimulation by intravenous (i.v.) injection of doses less than 40 micrograms/kg of the alpha 1-adrenoceptor blocking drug prazosin. However, prazosin (50-500 micrograms/kg i.v.) did not impair EDPs induced by preganglionic stimulation (0.5-64 Hz) or by injection of the nicotinic ganglion stimulant DMPP (50 micrograms/kg i.v.). Prazosin (50 micrograms/kg i.v.) inhibited EDPs induced by unilateral electrical stimulation (square wave pulses, 1 ms duration, 16 Hz, 2 s train length, 1 min intervals) of the spinal cord at C 1 in cats with an axotomy at the level of the medulla oblongata, thus indicating a spinal site of prazosin action and suggesting a permissive role of spinal catecholamines by activation of alpha 1-adrenoceptors. In spinal preparations pretreated with 250 micrograms/kg yohimbine i.v. to block inhibition by alpha 2 adrenoceptors of catecholamine reuptake, cocaine 2.5 mg/kg i.v. potentiated EDPs induced by spinal stimulation with 8 Hz. This effect could be antagonized by 50 micrograms/kg prazosin or 1000 micrograms/kg corynanthine i.v. In spinal preparations pretreatment with 5 mg/kg reserpine i.p. for depletion of catecholamines and 250 micrograms/kg yohimbine i.v., EDPs (16 Hz) were smaller than in undepleted preparations. Under these conditions injection of 100 micrograms/kg clonidine i.v. caused amplification of EDPs. This effect was antagonized by 50 micrograms/kg prazosin i.v. After i.v. pretreatment with 250 micrograms/kg yohimbine the i.v. injection of 2.5 mg/kg cocaine also potentiated EDPs which were induced by hypothalamic stimulation in intact cats. The results indicate that catecholaminergic neurons influence sudomotor activity by interaction with efferents of the cholinergic-sympathetic nervous system at the level of the spinal cord. Catecholamines seem to facilitate impulse transmission in non-catecholaminergic synapses by activation of alpha 1-adrenoceptors. PMID- 2990971 TI - GABAergic drugs and sexual behaviour in the male rat. AB - The GABAA agonists 3-amino-1-propanesulfonic acid and THIP reduced sexual behaviour in male rats only at relatively high doses, whereas baclofen produced an almost complete inhibition at a low dose (2.5 mg/kg). The GABA transaminase inhibitor aminooxyacetic acid had no effects, while gamma-acetylenic GABA produced a slight inhibition of sexual behaviour. The GABAA antagonist bicuculline had no effect. When THIP was administered concurrently with bicuculline, the former drug was potentiated. Therefore it is concluded that the GABAA receptor is not responsible for the inhibitory actions of THIP, and since baclofen was the most potent drug with regard to effects on sexual behaviour, it is suggested that the GABAB rather than the GABAA receptor is involved in the control of that behaviour. The slight effects of the transaminase inhibitors and the lack of effect of bicuculline suggest that the GABAergic neurons participating in the control of sexual activity are not tonically active. Finally, data are presented showing that the effects of GABAergic drugs on sexual behaviour are probably independent from those on locomotor activity. PMID- 2990973 TI - Horizontal transmission of the mouse mammary tumor virus in cage mates of the same and opposite sex of low and high mammary cancer strain mice. AB - Horizontal transmission of mouse mammary tumor virus (MTV) was investigated in cage mates of the same and opposite sex of low (BALB/c) and high mammary cancer strains (DD/Tbr, SHN and GR). By MTVp27 and MTVgp52 radioimmunoassay, MTV antigen expression was found in the salivary glands, mammary glands and secondary male genital organs of the MTV-free BALB/c strain. Infectivity and oncogenicity were also found in DDf or BALB/c mice by inoculating extracts of salivary gland and/or seminal vesicle in high mammary cancer strains. It is suggested that the primary source of the infectious agent in cases of caged animals of the same sex is saliva, while the primary source in cases of caged animals of the opposite sex is the seminal fluid, although additional infection through saliva cannot be ruled out in the latter case. PMID- 2990972 TI - Effects of PK 8165, a partial benzodiazepine receptor agonist, on cholecystokinin induced activation of hippocampal pyramidal neurons: a microiontophoretic study in the rat. AB - Benzodiazepines (BZD) have been reported to suppress cholecystokinin-8S (CCK-8S) induced activation. PK 8165, a ligand of BZD receptors, is an anxiolytic devoid of sedative and anticonvulsant effects. PK 8165, applied microiontophoretically or administered i.v. at low doses, suppressed CCK-8S-induced activation of hippocampal pyramidal neurons, whereas, at high doses it antagonized the effect of microiontophoretic applications of flurazepam. These results indicate that PK 8165 acts as a mixed agonist-antagonist at BZD receptors and suggest that the suppression of CCK-8S-induced activation by BZD might be related to their anxiolytic property rather than to their sedative or anticonvulsant activity. PMID- 2990974 TI - Diagnosis and localization of an insulinoma with inappropriate hypoglycemia in relation to the level of immunoreactive insulin using gel chromatographic separation. AB - We evaluated the possibility to diagnose the case of insulinoma using the combination of portal blood sampling and gel filtration techniques. The portal blood sampling showed 52 muU/ml of immunoreactive insulin (IRI) level at the closest splenic vein to the tumor, but the level should not be high enough to reasonalize the being of the tumor. The gel filtration pattern of IRI from the blood at the same point was clearly different from the other samples. Therefore, it could be useful for the diagnosis of insulinoma to combine percutaneous transhepatic portal blood sampling and gel filtration in such a case. PMID- 2990975 TI - The effect of long-term verapamil treatment on the secretion of cortisol and aldosterone in subjects with normal and high blood pressure. AB - Based on their previous observations of the stimulating action of verapamil on the secretory reserve of cortisol after one-week oral administration to healthy volunteers, the author investigated the effect of this calcium entry blocker on the adrenocortical function (cortisol and aldosterone secretion) during therapeutic administration to patients with mild arterial hypertension and normotensive patients suffering from Raynaud's syndrome. Verapamil, 3-4 X 80 mg per day by the oral route, did not lead to significant changes of cortisol levels at rest nor after ACTH stimulation when assessed at the end of the first month of treatment in both groups. However, normotensive subjects with Raynaud's syndrome showed a decline of ACTH stimulated cortisolaemia during the 3rd month (p less than 0.05) and 4th month (p less than 0.01) and a reduction of cortisol secretory reserve (delta cortisol) during the 3rd month of treatment (p less than 0.05). In aldosterone secretion of normotensives with Raynaud's syndrome no significant changes were observed. In hypertensive subjects, the decline of ACTH stimulated cortisolaemia at the end of the 3rd and 4th month was not significant and delta cortisol did not change. The initial aldosterone levels before and after ACTH and delta aldosterone before treatment of hypertensives was lower as compared with normotensives (p less than 0.01, p less than 0.01 and p less than 0.05, respectively), and increased gradually during treatment in the 4th month significantly (p less than 0.05, p less than 0.01 and p less than 0.01, respectively). The hypotensive effect of verapamil persisted throughout the treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990976 TI - In vitro effect of ritodrinum on cAMP concentration in human placentas of different gestational age. AB - Beta-sympathomimetic drugs are clinically widely used in the management of threatened premature labour. The existence of beta-adrenergic receptors has been demonstrated in the human placenta and, as the stimulation of beta adrenoreceptors is known to be mediated by cAMP, the purpose of this study was to compare the effect of Ritodrinum on cAMP concentration in placentas of different gestational age. The in vitro treatment with Ritodrinum induced a five fold increase in cAMP concentration in 6-8 week old placenta, three fold increase in midgestation, but was without effect in term placenta. This result suggest that beta-mimetics in pharmacological concentrations could modulate trophoblast cell activity through the cAMP-dependent system during the first half of gestation. PMID- 2990977 TI - Growth hormone, cyclic adenosine-3',5'-monophosphate and creatinine in relation to growth in the pig. AB - 28 male castrated pigs of the Land race, aged 90 to 110 days and of different growth were used as animal models. The parameters investigated were as follows: body weight at the time of investigation, density of carcass, creatinine excretion per 24 hours, glucose concentration in whole blood, the concentration of free fatty acids, growth hormone, fluorescence positive corticosteroids, cAMP and insulin in the blood plasma. The results show that there exists an obvious relation between the growth hormone concentration and growth progress in the pig, but it can be revealed only through an integrated growth hormone concentration. That is why the conclusion seems to be justified that the corticosteroid concentration in plasma, an integrated growth hormone concentration in plasma, the cAMP concentration in plasma and the amount of creatinine in urine within 24 hours are parameters which are related to the intensity and quality of growth in the pig. PMID- 2990978 TI - Effects of aging on the lipid order and composition of rat adipocyte ghosts. AB - An analysis of the cholesterol/phospholipid ratio of adipocyte ghosts from rat epididymal fat pads shows a significant increase with age (P less than 0.005). An attempt to correlate these changes with the order of the lipid matrix was made using the stearic acid spin label 2-(3-carboxypropyl)-4, 4-dimethyl-2-tridecyl-3 oxazolidinyloxyl [I(12,3)]. Although order was negatively correlated with temperature in preparations from both 6- and 24-month-old rats, no effect of age could be detected. PMID- 2990979 TI - Computer-assisted sequential quantitative analysis of gallium scans in pulmonary sarcoidosis. AB - Fifty-one sequential gallium citrate scans were performed in 22 patients with biopsy-proven sarcoidosis. A computer-assisted quantitative analysis of these scans was performed to obtain a gallium score. The changes in gallium score were correlated with changes in serum angiotensin converting enzyme (SACE) activity and objective changes in clinical status. There was a good concordance between changes in gallium score, SACE activity and clinical assessment in patients with sarcoidosis, and changes in gallium index were slightly superior to SACE index in assessing activity of sarcoidosis. PMID- 2990980 TI - Intravascular bronchioloalveolar tumour. AB - We report the clinical and histological characteristics of a case of intravascular bronchioloalveolar tumour, a rare multicentric pulmonary neoplasm of endothelial origin. This tumour affects predominantly women under 40 years of age and causes initially few symptoms. The disease may be found incidentally on a routine chest X ray. This tumour has a low metastasizing capacity and survival rate is quite high. PMID- 2990981 TI - Ultrastructural identification of somata and neural processes immunoreactive to antibodies against glutamic acid decarboxylase (GAD) in the dorsal lateral geniculate nucleus of the cat. AB - The cat dorsal lateral geniculate nucleus (LGN) was examined at the light- and electron-microscopic level after immunocytochemistry for GAD (the synthesizing enzyme of the inhibitory neurotransmitter GABA), to identify cells and processes with GAD-like immunoreactivity. GAD-positive perikarya were distributed throughout the A and C laminae, constituting a moderate proportion of cells in the LGN. Labeled cells were characterized by small size, scant cytoplasm, relatively large nuclei with common indentations, small mitochondria, few organelles and few strands of rough endoplasmic reticulum. Unlabeled cells were of large, medium and small size. GAD-positive terminals were identified as F1 and F2 types (Guillery's nomenclature) on the basis of their synaptic relations and ultrastructure. Labeled F2 terminals were postsynaptic to retinal (RLP) boutons and presynaptic to unlabeled dendrites in synaptic glomeruli. Labeled F1 terminals made synapses on unlabeled somata and dendrites, and on labeled dendrites and F2 terminals. Presumably, most labeled F1 terminals originate from GABAergic perigeniculate axons. Retinal (RLP) and cortico-geniculate (RSD) boutons remained unlabeled in the reactive zone. These terminals made synapses with labeled and unlabeled dendrites and with labeled F2 boutons. In conjunction with previous studies on GAD-positive cells in the perigeniculate nucleus, these results provide immunocytochemical and morphological evidence suggesting that the GABAergic intrinsic and extrinsic (perigeniculate) interneurons mediate the different inhibitory phenomena which occur in relay cells of the cat LGN. The ultrastructural features and synaptic relations of GABAergic cells and processes in the cat LGN are similar to those of equivalent neural elements in the LGN of rat and monkey, suggesting general principles of organization and morphology for GABAergic neurons in the thalamus of different mammals. PMID- 2990982 TI - Control of locomotion in marine mollusc--Clione limacina. V. Photoinactivation of efferent neurons. AB - Efferent neurons in isolated pedal ganglia of the pteropodial mollusc Clione limacina were filled with Lucifer Yellow through the wing nerves. Then the ganglia were illuminated with intense blue light which resulted in the complete inactivation of these neurons. After inactivation of efferent neurons, interneurons of the pedal ganglia continued to generate the locomotor rhythm. PMID- 2990983 TI - The cholinergic innervation of the visual thalamus: an EM immunocytochemical study. AB - In this study we demonstrate at the ultrastructural level that both the dorsal lateral geniculate nucleus (dLGN), the visual relay of the thalamus, and the perigeniculate nucleus (PGN), the visual segment of the thalamic reticular nucleus (TRN), are densely innervated by fibres with Choline-Acetyl-Transferase (ChAT) like immunoreactivity. These axons make synaptic contacts with interneurones considered to be inhibitory, both in the PGN and within the synaptic glomeruli of the dLGN. In addition, Chat positive terminals form intra- and extraglomerular synapses with dendrites thought to arise from relay cells. We interpret these results as evidence for direct cholinergic modulation of both relay cells and inhibitory interneurones. PMID- 2990984 TI - Quinolinic acid stimulates luteinizing hormone secretion in female rats: evidence for involvement of N-methyl-D-aspartate-preferring receptors. AB - Pharmacological evidence suggests that endogenous excitatory amino acid neurotransmitters stimulate luteinizing hormone (LH) secretion in neonatal and adult rats. Recent studies have identified quinolinic acid (QUIN), an endogenous brain and peripheral metabolite of tryptophan, as a potent agonist at N-methyl-D aspartate (NMDA)-preferring excitatory amino acid receptors. The present studies examined whether QUIN alters LH secretion in ovariectomized, estradiol-primed rats and whether such effects are mediated by specific amino acid receptor subtypes. In one experiment, animals received intracisternal injections of either quinolinic acid, N-methyl-DL-aspartate (NMA), aspartate (ASP), quisqualic acid (QA), or monosodium glutamate (GLU) five minutes prior to decapitation. In a second study, animals receiving central QUIN or NMA were treated simultaneously with either 2-amino-7-phosphonoheptanoic acid (APH) or kynurenic acid (KYA), both antagonists of NMDA-preferring receptors, or the quisqualate antagonist, glutamate diethyl ester (GDEE). Serum LH concentrations were measured by radioimmunoassay. Intracisternal administration of either QUIN or NMA resulted in an acute, dose-dependent increase of serum LH concentrations. Coadministration of APH blocked the effects of QUIN and NMA. QUIN stimulation of LH was also blocked by KYA, but not GDEE. Neither GLU nor ASP increased LH release, but QA did produce a small, significant elevation of LH. Light microscopic evaluation of brains showed no morphologic disturbance resulting from administration of these agents. The present results suggest that QUIN, or other endogenous ligands of NMDA-preferring receptors, may participate in the regulation of LH secretion in the adult female rat. PMID- 2990986 TI - Selective lesions of the fimbria and the fornix in the rat: differential effects on CA1 and dentate theta. AB - The effects of electrolytic lesions of the dorsal fornix and the dorsomedial fimbria on the CA1 and the dentate theta rhythms (theta s) recorded from the dorsal hippocampal formation were investigated in the ether-anesthetized rat. The results showed that (i) fornix lesions mainly affected CA1 theta, (ii) fimbrial lesions mainly affected dentate theta, and (iii) combined fornix-fimbria lesions suppressed both CA1 and dentate theta s. When considered in connection with other observations, these data suggest that the septal projections pacing the CA1 theta may course essentially within the dorsomedial fornix whereas those pacing the dentate theta may pass essentially within the dorsomedial fimbria. Moreover, our data provide new support for the hypothesis that at least two septohippocampal neural systems are anatomically and functionally independent and capable of controlling the theta activity of the dorsal hippocampal formation of the anesthetized rat. PMID- 2990985 TI - Quinolinic acid stimulates luteinizing hormone secretion through a serotonin dependent mechanism. AB - Previous results from this laboratory suggest that the tryptophan metabolite, quinolinic acid (QUIN), stimulates luteinizing hormone (LH) secretion in female rats, most likely through actions on NMDA-preferring excitatory amino acid receptors. The present experiments examined whether QUIN alters LH secretion through actions requiring intact catecholaminergic or serotonergic mechanisms. Each study examined the effects of intracisternal (i.c.) injections of 25 microliter acidic saline or saline containing QUIN (500 nmol) or the synthetic analogue, N-methyl-DL-aspartate (NMA, 500 nmol), into ovariectomized, estradiol benzoate-primed rats after pharmacologic disruption of monoaminergic neurotransmission. In each experiment, animals were decapitated 5 min after QUIN or NMA administration. Experiment 1 examined whether reduction in brain serotonin (5-HT) or of norepinephrine (NE) and epinephrine (E) alters the QUIN- or NMA induced stimulation of LH secretion. Rats were pretreated with the dopamine-beta hydroxylase inhibitor, FLA-63 (40 mg/kg 2 h prior). A second experiment examined the effects of the 5-HT antagonist, methysergide, on QUIN or NMA stimulation of LH secretion. Methysergide (15 mg/kg) was administered 30 min prior to experimentation. Experiment 3 examined whether selective destruction of raphe serotonergic neurons with the indoleamine neurotoxin, 5,7 dihydroxytryptamine (5,7 DHT), alters QUIN's stimulatory effects. In each study, serum LH concentrations were determined by radioimmunoassay. Hypothalamic catecholamine and 5-HT concentrations were measured by radioenzymatic assay and liquid chromatography with electrochemical detection, respectively. Depletion of brain 5 HT with PCPA significantly reduced the stimulation of LH secretion by QUIN, but not by NMA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2990987 TI - Evaluation of different GABA receptor agonists in the kindled amygdala seizure model in rats. AB - The effects of three specific GABA receptor agonists, muscimol, progabide, and gaboxadol, on kindled seizures were evaluated in amygdala-kindled rats. The only compound that exerted significant anticonvulsant effects at nonsedative doses was progabide. Thus, after i.p. administration of 100 mg/kg progabide, a significant increase in seizure latency and significant decreases in duration of motor seizures and amygdala afterdischarges were determined. A decrease in severity of motor seizures was found only after 200 mg/kg progabide which, however, gave rise to marked sedation and muscle relaxation. Muscimol and gaboxadol were almost inactive in attenuating kindled seizures even at doses that produced pronounced side effects. Assuming that the amygdala-kindled rat is a useful model of complex partial seizures with secondary generalization in the human, the data suggest that GABA receptor agonists are not effective against this type of epilepsy (muscimol, gaboxadol) or effective only at large doses (progabide). PMID- 2990988 TI - Estradiol increases choline acetyltransferase activity in specific basal forebrain nuclei and projection areas of female rats. AB - Administration of estradiol to gonadectomized female, but not male rats, is associated with increased activity of choline acetyltransferase in the medial aspect of the horizontal diagonal band nucleus, the frontal cortex, and CA1 of the dorsal hippocampus. Four other basal forebrain cholinergic nuclei did not show changes in choline acetyltransferase activity after estradiol. These data have implications for possible benefits of estradiol administration to patients with senile dementia of the Alzheimer's type. PMID- 2990989 TI - Plasmodium berghei: inhibitors of ornithine decarboxylase block exoerythrocytic schizogony. AB - DL-alpha-difluoromethylornithine and DL-alpha-monofluoromethyldehydroornithine methyl ester, inhibitors of ornithine decarboxylase, blocked exoerythrocytic schizogony of Plasmodium berghei in mice and in cultured human hepatoma cells. These effects were reversed by exogenous administration of the polyamine, spermidine. The antimalarial drug, primaquine, the side chain of which is structurally analogous to a natural polyamine, did not enhance the activity of alpha-difluoromethylornithine or alpha-monofluoromethyldehydroornithine methyl ester. These results extend previous observations that polyamines influence the malaria parasite's schizogony outside the red blood cell but not within it. PMID- 2990991 TI - Plasmodium falciparum: differentiation of isolates with DNA hybridization using antigen gene probes. AB - Chromosomal DNA was prepared from seven Plasmodium falciparum isolates that had been cultured in vitro and from a cloned P. falciparum line. The DNA was cleaved with restriction endonucleases, fractionated by agarose gel electrophoresis, blotted to nitrocellulose, and hybridized with a series of radioactively labeled DNA probes. The probes had been derived from cDNA clones encoding portions of P. falciparum antigens. Simple, reproducible band patterns that differed for many of the isolates were obtained. Parasite isolates collected from different continents could be readily distinguished, as could some but not all isolates collected from one restricted region of Papua New Guinea. Application of this technique for the identification and differentiation of parasite strains was explored. The patterns of hybridization observed were consistent with the proposition that blood stages of P. falciparum have a haploid genome. PMID- 2990990 TI - Schistosoma mansoni: cercarial eicosanoid production and penetration response inhibited by esculetin and ibuprofen. AB - Cercariae of Schistosoma mansoni produce a wide variety of eicosanoids when stimulated by 3.3 mM linoleate. High-performance liquid chromatography indicated that 10(-5) M esculetin dramatically decreased eicosanoid production by cercariae. Ibuprofen (10(-4) M) also decreased eicosanoid production, but to a lesser extent. These results were confirmed by radioimmunoassay using time-dose curves for esculetin and time curves for ibuprofen. The results reported here indicated that, for cercariae, ibuprofen was neither a specific cyclo-oxygenase inhibitor, as has been reported for platelet and endothelial cells, nor was esculetin a specific inhibitor of lipoxygenase, as has been reported for platelets and mastocytoma cells. Rather, both drugs inhibited cyclo-oxygenase and lipoxygenase enzyme systems. Further, the data indicated that two forms of cyclo oxygenase exist in cercariae (isozymes?), one initiating the conversion of gamma dihomolinolenate into the 1-series prostaglandins and another acting on arachidonate forming the 2-series prostaglandins. The cyclo-oxygenase acting on arachidonate has a greater sensitivity to both ibuprofen and esculetin than the enzyme acting on gamma-dihomolinolenate. Cercarial lipoxygenases also varied greatly in their sensitivity to ibuprofen. PMID- 2990992 TI - Elastase, collagenase and the radial elastic properties of arteries. AB - Studies were performed on 203 pairs of dog carotid arteries subjected to unidirectional radial compression. Treatment with 80 U/ml purified elastase for 90 min decreased radial stress, but treatment with 640 U/ml collagenase for 90 min did not. These data suggest that elastin, but not collagen, contributes to wall resistance to radial compression. PMID- 2990993 TI - Failure of calcium to stimulate Na,K-ATPase in the presence of EDTA. AB - The effect of calcium on Na,K-ATPase activity of rat brain homogenates and its modification by the chelating agent EDTA has been investigated. In the absence of EDTA, free calcium (approximately 10(-6) mol/l) stimulates Na,K-ATPase activity; in the presence of EDTA the same concentration of free calcium is without effect on the enzyme. In the absence of EDTA the stimulation by calcium of Na,K-ATPase activity is enhanced by the additional presence of calmodulin but in the presence of EDTA, even when calmodulin is added to excess, calcium still fails to stimulate the enzyme. The possibility that EDTA interferes with an interaction between a calcium-calmodulin complex and Na,K-ATPase is discussed. PMID- 2990994 TI - Energy metabolism and transduction in smooth muscle. AB - Early investigations into the nature of the coupling between energy transduction and metabolism in smooth muscle, particularly from the laboratories of Bulbring and Lundholm, suggested that specific metabolic pathways could independently supply energy for ion transport and actin-myosin interactions. Subsequent work has solidified the concept that oxidative phosphorylation is specifically coupled to tension generation and maintenance, whereas, aerobic glycolysis is not only a vital characteristic of smooth muscle metabolism, but also is likely to be independently coupled to Na-K transport at the plasmalemma. The independence of oxidative and glycolytic metabolism is reflected as a compartmentation of carbohydrate metabolism in the porcine carotid artery. The coupling of these independent metabolic pathways with specific energy utilizing processes, indicates a means by which energy production and transduction can be closely and efficiently regulated. The coupling of glycogenolysis to mitochondrial respiration may have evolved as a direct response to the energetic needs of VSM. That is, the large glycogenolytic response in the initial minutes of stimulation may be necessary to maximize the cellular production of ATP during the presteady state. Likewise, the coupling between aerobic glycolysis and Na-K transport indicates a sensitive and efficient means of coordinating energy metabolism with ion transport at the membrane level. Additionally, the regulation of substrate supply, i.e. glucose transport, also may be closely coordinated with changes in ion transport. One may speculate that alterations in the microenvironment of each compartment can independently regulate intermediary metabolism and therefore allow the cell to quickly and efficiently respond to localized stimuli. Thus, stimulation of Na-K transport could effectively regulate energy production at the membrane level without mobilizing or competing with the energy transduction of other cellular processes. This compartmentation of energy utilization may be highly advantageous, since oxidative metabolism is closely coordinated with mechanical activity and therefore regulation of blood flow. Future investigations will attempt to elucidate which intracellular signals which are responsible for the regulation of these functionally independent compartments of energy metabolism and transduction in VSM. In more general terms, our findings provide a basis from which future questions concerning the regulation of cellular metabolism must be directed. The cellular cytoplasm can no longer be envisioned as a homogeneous compartment, but rather a complex array of functional subcompartments which may be individual PMID- 2990995 TI - Ca2+ in biological systems. PMID- 2990996 TI - Calcium and sodium distribution and movements in smooth muscle. AB - Electron probe microanalysis (EPMA) has been used to study the subcellular distribution of Ca, Na, K, Cl, and Mg in smooth muscle. The EPMA results indicate that the sarcoplasmic reticulum (SR) is the major intracellular source and sink of activator Ca: norepinephrine decreases the Ca content of the junctional SR in portal vein smooth muscle. Mitochondria do not play a significant role in regulating cytoplasmic free Ca2+, but mitochondrial Ca content can be altered to a degree compatible with suggestions that fluctuations in matrix Ca contribute to the control of mitochondrial metabolism. The rise in total cytoplasmic Ca during a maintained, maximal contraction is very much greater than the rise in free Ca2+, and is probably in excess of the known binding sites available on calmodulin and myosin. Cell Ca is not increased in normal cells that are Na loaded. The non-Donnan distribution of Cl is not due to compartmentalization, but reflects high cytoplasmic Cl. Na-loading of smooth muscle in K-free solutions is temperature dependent, and may exhibit cellular heterogeneity undetected by conventional techniques. The total cell Mg is equivalent to approximately 12 mM, and less than 50% of it can be accounted for by binding to ATP and to actin. Mitochondrial monovalent cations in smooth muscle are relatively rapidly exchangeable. PMID- 2990998 TI - Modulation of calcium sensitivity in guinea pig taenia coli: skinned fiber studies. AB - In summary then, skinned fiber experiments have provided evidence showing that the relationship between force development and free calcium ion concentration may be variable. The cyclic-nucleotides c-GMP (cf 38) and c-AMP in particular might be important in modulating calcium sensitivity of skinned fibers possibly via an alteration of myosin phosphorylation. Additionally, the coupling between myosin phosphorylation and the ATPase activity of actomyosin or tension generation may itself be modulated in actomyosin contractile systems. The recognition of the physiological relevance of these modulating mechanisms however must await experiments in which the relationship between force development, free calcium ion concentration and myosin phosphorylation is studied in intact fibers. With the advent of more sophisticated calcium measuring techniques, such information is now available. PMID- 2990999 TI - A route to the total synthesis of iso-LTB4 system. AB - The convergent synthesis of the iso-Leukotriene B4 (XVI) (as diastereoisomeric mixture with the desired geometry), by Wittig's condensation of the two precursor synthons at 6 (VI) and 14 C (XIV), starting from the common cyclic acrylaldehyde dimer (I), is reported. PMID- 2990997 TI - Calcium-induced calcium release mechanism in vascular smooth muscles--assessments based on contractions evoked in intact and saponin-treated skinned muscles. AB - This article was concerned with the role of Ca in triggering the contraction in vascular smooth muscles. Whenever Ca influx is activated, this Ca does not directly activate the contractile proteins, but rather triggers the release of Ca from the SR to activate calmodulin. This release of Ca by Ca is dependent on the amount of Ca stored within the cells. Voltage dependent Ca influx activated by excess concentrations of K, electrical depolarization and Ca spikes is required to produce the contraction through activation of the Ca-induced Ca release mechanism. The elucidation of the contribution of the P-I response for Ca mobilization through activation of receptors under physiological conditions hopefully will lend support to our hypothesis. PMID- 2991000 TI - Synthesis of a hydrolase for the membrane-form variant surface glycoprotein is repressed during transformation of Trypanosoma brucei. AB - A membrane-bound phospholipase C-like hydrolase present in lysates of bloodstream forms of Trypanosoma brucei rapidly converts the membrane form of the variant surface protein to the soluble form and 1,2-dimyristoylglycerol [(1985) M.A.J. Ferguson et al. J. Biol. Chem., 260, 4963-4968]. The hydrolase is inhibited by p chloromercuribenzenesulfonate. The synthesis of the enzyme is rapidly repressed upon differentiation of bloodstream forms to procyclic cells and the enzyme activity declines to an undetectable level during subsequent growth of procyclic forms. PMID- 2991001 TI - The reduction of cytochrome c oxidase by carbon monoxide. AB - The rate of formation of the mixed-valence state of cytochrome c oxidase on incubation with carbon monoxide is strongly dependent on pH, supporting the concept that CO itself is the reducing agent. The reaction is biphasic due to the presence of two different enzyme forms in the resting oxidase. The kinetics of the reaction with the major enzyme form suggests an initial rapid binding of CO in a heme pocket of the oxidized enzyme and a subsequent slow intramolecular electron transfer to the cytochrome alpha 3-CuB site. Cytochrome alpha and CuA are not reduced even on prolonged incubation. PMID- 2991002 TI - The amiloride sensitive Na+/H+ antiport in guinea pig pancreatic acini. Characterization and stimulation by caerulein. AB - Amiloride and analogs decrease the initial rate of 22Na+ uptake by dispersed acini from guinea pig pancreas in a dose-dependent manner. The initial rate of amiloride-sensitive 22Na+ uptake depends on external Na+ and H+ concentrations and on internal pH. These results provide evidence for the existence of a Na+/H+ antiport in pancreatic acinar cells. Caerulein, a cholecystokinin analog, stimulates the activity of the Na+/H+ antiport. PMID- 2991003 TI - Papain digestion of the subunits of (Na,K)-ATPase. AB - Membrane-bound (Na,K)-ATPases were exposed to limited papain digestion. We could not find the active (Na,K)-ATPase lacking glycoprotein subunit for the enzymes from three different sources (outer medulla of dog kidney, electric organs of Narke japonica and larvae of Artemia salina). It seemed unlikely that the glycoprotein subunit was selectively removed from (Na,K)-ATPase by papain digestion. PMID- 2991004 TI - On the prosthetic group(s) of component II from nitrogenase. EPR of the Fe protein from Azotobacter vinelandii. AB - The EPR spectrum of the reduced Fe-protein from nitrogenase has been reinvestigated. The dependences on temperature, microwave power, and microwave frequency all suggest that the observed signal represents a magnetically isolated [4Fe-4S]1+(2+;1+) cluster. Also, the signal can be simulated assuming a simple, g strained S = 1/2 system. However, the integrated intensity amounts to no more than 0.2 spins per protein molecule. It is, therefore, impossible that Fe-protein preparations contain a single type of [4Fe-4S] cluster. PMID- 2991005 TI - Thermodynamic limits to the stoichiometry of H+ pumping by mitochondrial cytochrome oxidase. AB - H+/O stoichiometries of 0, 2 and 4 have been proposed for cytochrome oxidase. Here we show that a stoichiometry of 4 is thermodynamically impossible for rat liver cytochrome oxidase under normal conditions. PMID- 2991006 TI - A role for adrenaline and calmodulin in modulating cyclic AMP levels during the lactogenic cycle. AB - The effect on lactose production of several external modulators of intracellular cyclic AMP was studied in rat mammary gland tissue slices and explants. Adrenaline, a beta-adrenergic receptor effector, forskolin, a direct adenylate cyclase activator and fluphenazine, a calmodulin inhibitor, all produced an increase in the intracellular level of cyclic AMP and a concomitant inhibition of lactose production. These results suggest a role for adrenaline and calmodulin in modulating cyclic AMP levels in mammary tissue during the lactogenic cycle. PMID- 2991007 TI - Three-dimensional structure of renal Na,K-ATPase determined by electron microscopy of membrane crystals. AB - The three-dimensional structure of Na,K-ATPase was determined by electron microscopy and image processing. Tilt series of negatively stained membrane crystals were recorded. The projections were analyzed by Fourier methods and the data combined to a 3-D model. The unit cell contains two rod-shaped stain deficient regions interpreted as alpha beta-promoters of Na,K-ATPase. The rods are related by dyad axes oriented perpendicular to the membrane. Outside the lipid bilayer the rods contact different protein units on the two sides of the membrane. PMID- 2991008 TI - The site of cyclic AMP-dependent protein kinase catalyzed phosphorylation of cytochrome P-450 LM2. AB - The phenobarbital-inducible form of cytochrome P-450 purified from rabbit liver microsomes is phosphorylated by cAMP-dependent protein kinase at a single site, the serine residue in position 128 of the amino acid sequence. The serine is located in a characteristic recognition sequence for cAMP-dependent protein kinase and is part of a primary structure which is conserved during evolution, present also in phenobarbital-inducible rat cytochrome and cytochrome P-450 CAM from Pseudomonas putida. The contribution of these findings to our understanding of the structure and membrane topology of cytochrome P-450 LM2 and its turnover regulated by phosphorylation is discussed. PMID- 2991009 TI - Reversible interaction between Ca2+-activated neutral protease (CANP) and its endogenous inhibitor. AB - The interaction between the Ca2+-activated neutral protease (CANP) and its endogenous inhibitor was analyzed. The interaction was completely reversible, and both CANP and the inhibitor regained full activity after dissociation of their complex. PMID- 2991010 TI - Collagenase in mineralized tissues of human teeth. AB - A collagenase cleaving native type I [14C]collagen but inactive against the synthetic substrate Pz-Pro-Leu-Gly-Pro-D-Arg was extracted from mineralized human dental tissue. The enzyme specifically degrades native collagen into characteristic products (3/4) and (1/4). Its apparent molecular mass of 68 kDa is relatively high in comparison with collagenases from other oral tissues. The enzyme is a metalloproteinase inhibited by low concentrations of the chelating agents EDTA, 1, 10-phenanthroline, alpha alpha'-dipyridyl, and not affected by diisopropylfluorophosphate, soybean trypsin inhibitor, and p chloromercuribenzoate. It is stable to lyophilization and can be stored at-20 degrees C for at least 6 months. PMID- 2991011 TI - Parthenogenetic activation decreases the polyphosphoinositide content of frog eggs. AB - Polyphosphoinositides were quantified in metaphase II-arrested eggs of the amphibian Xenopus laevis and 8-10 min later in eggs activated by pricking. The content of phosphatidylinositol 4,5-biphosphate (PIP2) was remarkably high in metaphase II-arrested eggs with respect to that of phosphatidylinositol 4 phosphate (PIP). It was found to drop dramatically at activation. In contrast PIP content did not change significantly. PMID- 2991012 TI - Epidermal growth factor stimulated protein kinase shows similar activity in liver of senescent and adult mice. AB - It was recently reported [(1983) Nature 306, 617-620] that tyrosine protein kinase activity associated with EGF receptor was absent from senescent human cultured fibroblasts, which are known to have the same number of receptors as young human cultured fibroblasts. We have measured in both adult and senescent C57 black mice the number of EGF receptors, the activity of their associated tyrosine kinase and the activity of the protein phosphatase which dephosphorylates the EGF receptor. We found our results in both groups of animals to be similar which indicate that the observations made in cultured fibroblasts cannot be generalized to all mammalian tissues. PMID- 2991013 TI - The phorbol ester TPA prevents the expression of both glucagon desensitisation and the glucagon-mediated block of insulin stimulation of the peripheral plasma membrane cyclic AMP phosphodiesterase in rat hepatocytes. AB - The phorbol ester TPA (12-O-tetradecanoyl phorbol-13-acetate) causes a dose dependent inhibition of the glucagon-stimulated adenylate cyclase activity expressed in plasma membranes isolated from TPA-treated hepatocytes. However, no observable inhibitory effect of TPA on adenylate cyclase activity was observed in cells which had been exposed to glucagon for 5 min, prior to isolation, to desensitise adenylate cyclase. The degree of inhibition of adenylate cyclase elicited by both glucagon desensitisation and TPA treatment of hepatocytes was identical. Pre-treatment of hepatocytes with TPA was also found to prevent glucagon from blocking insulin's activation of the peripheral plasma membrane cyclic AMP phosphodiesterase in intact hepatocytes. TPA treatment also inhibited the ability of cholera toxin to activate the peripheral cyclic AMP phosphodiesterase in intact hepatocytes. It is suggested that in these particular instances TPA and glucagon elicit mutually exclusive processes rather than TPA mimicking glucagon desensitisation per se. PMID- 2991015 TI - Degradation of alpha-actinin during Ca2+-sensitive proteolysis of myofibrils. AB - The noted loss of alpha-actinin from the Z-line of myofibrils during post-mortem autolysis, probably following the action of calcium-activated protease, has previously been attributed to its release without degradation. This report shows that in isolated myofibrils alpha-actinin is proteolysed in a Ca2+-sensitive manner presumably via the action of calcium-activated protease. PMID- 2991014 TI - Effect of ascorbic acid on the production of singlet oxygen by purified human myeloperoxidase. AB - We have previously studied purified human myeloperoxidase-hydrogen peroxide halide ion systems as models of possible singlet oxygen production by granulocytes. While myeloperoxidase could efficiently produce singlet oxygen, the yield of singlet oxygen at a physiological pH with Cl- was very small due to enzyme inactivation. In that Bolscher et al. [(1984) Biochim. Biophys. Acta 784, 189-191] observed that micromolar concentrations of ascorbic acid prevented inactivation of myeloperoxidase and increased the production of hypochlorous acid, we examined whether ascorbic acid would augment singlet oxygen production by the myeloperoxidase-hydrogen peroxide-halide ion systems. Ascorbic acid, however, fails to increase the singlet oxygen yield, suggesting that it does not augment singlet oxygen production in the intact granulocyte by a myeloperoxidase dependent mechanism. PMID- 2991016 TI - Molecular cloning of a full-length cDNA encoding the hemagglutinin-neuraminidase glycoprotein of Sendai virus. AB - We cloned a full-length complementary DNA for the hemagglutinin-neuraminidase (HN) mRNA of Sendai virus (HVJ) using a synthetic 27-mer as a probe. Nucleotide sequence analysis showed that there is a long open reading frame on the mRNA that encodes a protein of 575 amino acids. The deduced amino acid sequence indicated that only one hydrophobic region sufficiently long to anchor the protein in the membrane and located near the N-terminus (amino acids 35-60). It is suggested that HN protein is oriented with its N-terminus inside the membrane. PMID- 2991017 TI - Comparison of genomes of closely related phages phi 29, phi 15 and PZA using a rapid method of parallel physical mapping. AB - The DNAs of phages phi 29, phi 15 and PZA of Bacillus subtilis were analysed with restriction enzymes EcoRI, HpaI and HindIII. A method was used which permits parallel physical mapping of all three phages, from both ends of their linear genomes. The method is based on transfer of partially digested DNA to DBM paper and sequential hybridization with labelled terminal fragments. It follows from the comparison of the physical maps that phages phi 29, phi 15 and PZA are closely related and that they probably have arisen from a common ancestor by accumulation of point mutations. PMID- 2991018 TI - Osmotic effects on bacterial transport and energetics. AB - Paracoccus denitrificans suspended in media containing 20-300 mM NaCl swelled progressively as the salt concentration was decreased. The increase in intracellular water volume was accompanied by an enhancement of respiration and a stimulation of the rates of net potassium and alpha-aminoisobutyric acid accumulation. It is postulated that influx of water and consequent lowering of intracellular solute concentration trigger transport mechanisms which are destined to restore the original ion and metabolite balance. Since a number of transport reactions operate against the electrochemical gradient of their substrates, energy utilization increases. The increased ATP usage and lowering of [ATP] stimulates the activity of the respiratory chain and increases oxygen uptake and energy production. PMID- 2991020 TI - Replacement of Mn(III) with Cu(II) in Bacillus stearothermophilus superoxide dismutase. Similarity of the active site to the zinc site of copper/zinc superoxide dismutase. AB - Copper(II) was substituted for manganese(III) in Bacillus stearothermophilus Mn superoxide dismutase. The (EPR) spectrum of the Cu(II) is distinctly rhombic, overlapping that of Cu(II) replacing zinc in copper/zinc superoxide dismutase. The copper-cyanide complex of the bacterial enzyme gives an EPR spectrum nearly identical to the cyanide-copper complex of the Cu/Zn enzyme, indicating three nitrogens as metal ligands. These results support the suggestion that metal coordination in B. stearothermophilus Mn-dismutase is a tetrahedral arrangement of three imidazoles and a carboxylate group and indicate copper as a good spectroscopic probe for studying the active site of Mn- and Fe-superoxide dismutases. PMID- 2991019 TI - 3 beta, 17 beta-hydroxysteroid dehydrogenase of Pseudomonas testosteroni. Kinetic evidence for the bifunctional activity at a common catalytic site. AB - 3 beta, 17 beta-Hydroxysteroid dehydrogenase (3 beta 17 beta HSDH) is an NAD dependent dehydrogenase which has a double specificity for the 3- and 17 positions on the steroid skeleton. When dehydroepiandrosterone (DHEA) is used as steroid substrate, and the assay coupled with ketosteroid-isomerase, the two reactions occur alternately and each reaction on the 3-position produces a chromophoric molecule. These two reactions can follow one another without dissociation of the coenzyme from the enzyme binding site. This is confirmed by competition experiments with another dehydrogenase. PMID- 2991021 TI - Adrenocorticotropic hormone testing in idiopathic hirsutism and polycystic ovarian disease: a test of limited usefulness. AB - The plasma 17 alpha-hydroxyprogesterone (17-OHP) concentration was determined in the basal state and 60 minutes after cosyntropin, 0.25 mg, in 139 patients with idiopathic hirsutism (IH) and polycystic ovarian disease (PCOD). Although there was an increased response of 17-OHP in subjects with PCOD when compared with IH subjects, in no instance was stimulated 17-OHP abnormal in the presence of normal basal 17-OHP. Two subjects with 21-hydroxylase (21-OH) deficiency were discovered; both demonstrated elevated basal levels of 17-OHP. We therefore conclude that routine adrenocorticotropic hormone testing is not a useful tool in detecting 21-OH deficiency in hyperandrogenic women. PMID- 2991023 TI - Pregnancy prevention by intravaginal delivery of a progesterone antagonist: RU486 tampon for menstrual induction and absorption. AB - A progesterone antagonist (RU486) was administered to macaque monkeys by an intravaginal tampon for the achievement of both menstrual induction and absorption. The study was conducted in three parts: (1) postcoital treatment beginning 3 days after potential fertilization; (2) treatment on extended cycle days 34 to 37 after a "missed" menstrual flow after mating; and (3) luteal phase (cycle days 22 to 25) treatment in noncoital cycles. Our findings demonstrate pregnancy prevention and/or interruption when these treatments are employed. PMID- 2991022 TI - Cervicovaginal peroxidases: sex hormone control and potential clinical uses. AB - Thirty-one normal women were studied daily in 41 cycles. Venous blood samples were taken for measurements of luteinizing hormone (LH), estradiol (E2), and progesterone (P), and vaginal examinations were done to obtain cervical mucus and vaginal fluid. The specific activity of guaiacol peroxidase (GP), extracted from cervicovaginal secretions with 0.5 M CaCl2, was determined in the vaginal samples. In the follicular phase, from day -7 to day 0 (the LH +1 day, when ovulation presumably occurred), there was a strong negative correlation between GP and the rising E2 (r = -0.94). On days 1 to 10 after ovulation, there was a strong positive correlation between GP and P (r = 0.84). In nine ovulatory cycles in which P levels did not exceed 8 ng/ml on any day, indicating possible luteal phase inadequacy, there were significantly lower GP levels than in another 32 ovulatory cycles with higher P (P = 0.04). These results suggest that (1) at midcycle, E2 seems to "down-regulate" the GP specific activity; and (2) in the luteal phase, serum P levels parallel those of GP activity, even in the presence of high luteal E2. GP activity profiles during the menstrual cycle can be used to define the fertile period, may prove useful in diagnosing pregnancy, and may be a simple, convenient test for an inadequate corpus luteum. PMID- 2991024 TI - [Effect of dorsal root section on synaptic potentials of frog spinal cord motor neurons]. PMID- 2991025 TI - Isolation and characterization of cell clones producing various amounts of bovine leukosis virus. AB - Several single-cell clones were isolated from lamb kidney cell line (FLK) persistently infected with bovine leukosis virus (BLV). The clones differed in the amount of produced virus, some clones were highly virus productive, others produced less virus. The restriction analysis of cell DNA from clones revealed the existence of several numbers of integrated BLV-DNA proviruses in various cell clones. In some cell clones the unintegrated proviral DNA was observed. The proviral BLV sequences were found to be integrated in reiterated DNA in some cell clones. There was no apparent relationship between the number of integrated BLV proviruses and the extent of virus production. The origin of the different numbers of integrated BLV proviruses in various cell clones is discussed. PMID- 2991026 TI - Molecular cloning of integrated proviral DNA of bovine leukaemia virus from virus producing foetal lamb kidney cells. AB - Over 90% of the total viral information together with the adjacent 5' cellular sequences were cloned in the lambdoid spi selection vector lambda-2558 by taking advantage of the unique EcoRI restriction site very close to the 3' long terminal repeat. Of the fifteen isolated recombinant phages with viral inserts, one has been propagated and its DNA isolated and subjected to preliminary restriction endonuclease analysis. The proviral insert has been found to be almost identical to the unintegrated proviral DNA reported by Kashmiri et al. (1984). PMID- 2991027 TI - Human genome: proto-oncogenes and proretroviruses. AB - A brief review of the studies undertaken at the Laboratory for Molecular Bases of Oncogenesis (Institute of Molecular Biology, Moscow) till middle of 1984 is presented. The human genome contains multiple dispersed nucleotide sequences related to the proto-oncogene mos and to proretroviral sequences in tight juxtaposition to each other. From sequencing appropriate cloned fragments of human DNA in phage and plasmid vectors it follows that one of these regions, NV 1, is a pseudogene of proto-mos with partial duplications and two Alu elements intervening its coding sequence, and the other, CL-1, seems to be also a mos related gene with a deletion of the internal part of the structural gene. CL-1 is flanked by a proretroviral-like sequence including tRNAiMet binding site and U5 (part of the long terminal repeat). The proretroviral-like sequences are transcribed in 21-35S poly(A)+RNA abundant in normal and malignant human cells. Two hypotheses are proposed: endogenous retroviruses take part in amplification of at least some proto-oncogenes; proto-oncogenes are inactivated via insertion of movable genetic elements and conversion into pseudogenes. Potential oncogenicity of a normal human genome undergoes two controversial influences: it increases due to proto-oncogene amplification and decreases due to inactivation of some of them. PMID- 2991028 TI - Isolation of two transforming viruses from sarcomas obtained in chickens inoculated intraembryonally with a transformation defective mutant of Prague strain Rous sarcoma virus. AB - Sarcomas appeared after long latency with a frequency of about 2% in Brown Leghorn and (CB X IC)F1 chickens after intraembryonic and neonatal inoculation of transformation-defective mutants of ASVs subgroup C. Only the freshly isolated td mutants, td daPR-C and td daPR-C morphf, exhibited the tumorigenic activity, whereas the standard td mutants induced no sarcomas. Two (862 and 2257) out of four tumours could be transplanted in young chickens and produced low titres of transforming virus. The other two tumours were not transplantable and devoid of any transforming virus. Viruses 862 and 2257 are clearly defective in replication, virus 2257 cannot be complemented efficiently by any helper virus in vitro. The low titre of virus 2257 is not caused by interference with a helper virus of the same subgroup specificity. Both viruses are highly tumorigenic in vivo. PMID- 2991029 TI - Identification of transformation-specific proteins synthesized in cryptovirogenic mammalian cells. AB - The cryptovirogenic PR-RSH-19 (H-19) hamster cell line, derived originally from a tumour induced by the virus rescued from XC cells by transfection, was studied. The pp60v-src protein was detected by immunoprecipitation with sera from tumour bearing rabbits (TBR sera) from lysates of H-19 cells. A closely related protein p60 was precipitated by antiserum obtained from hamsters with tumours induced by H-19 cells. The pp60v-src of H-19 cells possesses an associated kinase activity. These results are consistent with the notion that the detected pp60v-src protein is a product of the cryptic sequences in H-19 cells. PMID- 2991030 TI - Amino acid sequence homology between protein products of oncogenes and hormones (v-myc--gastrin and oxytocin; v-sis--secretin). AB - Computer comparisons of amino acid sequences from 8 retroviral oncogenes and 26 protein hormones were done with respect to structure similarity and so-called uninterrupted structure similarity. Sequence homology was found between v-myc, the transforming protein of MC29 virus, and human gastrin and oxytocin on the one hand, and between v-sis, the transforming protein of simian sarcoma virus, and secretin on the other. PMID- 2991032 TI - Effect of the expression of an endogenous viral gene on the growth of tumours induced by Rous sarcoma virus in chickens. AB - The endogenous group-specific antigen of avian retroviruses has been detected in feather pulp of chickens of the inbred CB line (WL breed). Expression of the gs antigen has been demonstrated by ELISA, whereas the classical methods have classified the CB line as gs-. The number of gs+ chickens decreased with increasing age after hatching, and all chickens were gs- already at 20 weeks of age. The longer duration of gs antigen expression in line CB chickens had an adverse effect on their ability to regress Rous sarcomas. Chickens from a closed flock of the BL breed that segregated to gs+ and gs- also significantly differed in the ability to regress Rous sarcomas. PMID- 2991031 TI - Partial sequence homology between retroviruses of various species in the C terminal part of the env gene. AB - We determined the nucleotide sequence of a molecularly cloned cDNA from the endogenous cat virus RD114. Comparison of the nucleotide sequence and the deduced amino acid sequence, respectively, with other known sequences of retroviruses revealed two conserved domains in the viral transmembrane protein, which we relate to structural features of the protein. PMID- 2991033 TI - [Establishment of anti-thyroglobulin antibody-producing cell line by EB virus transformation]. AB - We established an anti-thyroglobulin antibody-producing cell line using the peripheral lymphocytes from a patient with chronic thyroiditis. The method was based on preselection by "panning", transformation by EB virus and twice clonings by "limiting dilution". The cells of the cloned cell line (Yo3CTX10) had neither E-receptor nor IgG X Fc-receptor but had C3-receptor as shown by rosetting. We could not detect surface immunoglobulin, but we could detect cytoplasmic immunoglobulin (IgG lambda) with FITC-stainings. This cell line has continuously secreted anti-thyroglobulin antibody for 8 months, which was IgG lambda as shown by immunoelectrophoresis and solid phase radioimmunoassay. The pattern obtained using the purified antibody showed two sharp bands (H, L-chains) in SDS polyacrylamide gel electrophoresis, but it showed six sharp bands in isoelectric focusing electrophoresis. Thus we could establish an oligoclonal cell line producing IgG lambda anti-thyroglobulin antibody. PMID- 2991034 TI - Mechanism of action of cholecystokinin: a not atypical brain-gut peptide. AB - Brain-gut peptide action has been best studied in certain target cells of the gastrointestinal tract such as isolated pancreatic acini. Cholecystokinin (CCK) activation of pancreatic digestive enzyme secretion is initiated by specific receptors present in the basolateral membrane of the acinar cell. These receptors are highly selective for CCK and readily discriminate it from the homologous peptide, gastrin. Studies covalently crosslinking 125I-CCK to its receptor have revealed a binding glycoprotein subunit of Mr = 76,000 attached by a disulfide bridge to a Mr = 40,000 nonbinding subunit. Receptor occupancy leads to phosphotidylinositide breakdown and Ca2+ mobilization. Recent studies with the fluorescent chelate probe Quin-2 have shown that CCK increases cytosolic Ca2+ from a basal level of 100 nM to 500 approximately 1000 nM. The effects of Ca2+, and diacylglycerol produced by the breakdown of phosphoinositides, are believed mediated by activation of a group of protein kinases and phosphatases. CCK in the brain is present in neurons and is released from nerve endings by depolarization. The cellular mechanism of action of CCK, however, is essentially unknown. CCK application excites certain neurons but attempts to demonstrate effects on ion fluxes, phospholipid metabolism and protein phosphorylation have been negative to date. A possible explanations is provided by the finding that the brain CCK receptor shows differences in binding specificity from peripheral CCK receptors. Moreover, crosslinking studies reveal a single binding protein of Mr = 51,000. Thus, CCK may act differently in the brain and pancreas. PMID- 2991035 TI - [The acute effects of the new angiotensin I-converting enzyme inhibitor, enalapril maleate, on blood pressure, plasma renin, aldosterone and kinins in hypertensive patients]. AB - The acute antihypertensive effect of a new long-acting oral angiotensin I converting enzyme (ACE) inhibitor, enalapril maleate, was assessed in 20 hypertensive patients, of whom 14 had essential hypertension, 4 had renovascular hypertension, one had hypertension associated with chronic renal failure, and one had primary aldosteronism. Enalapril maleate significantly lowered the blood pressure in either low-renin or normal- and high-renin hypertensives. There was a significant correlation for all patients as a group between the pretreatment levels of serum ACE activity and the reduction in mean blood pressure (r = 0.454, p less than 0.05, n = 20) 2 h after drug administration. The serum ACE activity decreased maximally 3 to 4 hours after drug administration and did not return to baseline levels within 24 h. There was a significant correlation between the reduction in mean blood pressure and changes in ACE activity 90 min and 2 h after drug administration, respectively, for all patients as a group (r = 0.495, p less than 0.05, n = 20, at 90 min; r = 0.508, p less than 0.05, n = 20, at 2 h). The plasma renin activity (PRA) significantly increased in normal- and high-renin hypertensives but not in low-renin hypertensives. There was a close correlation between the reduction in mean blood pressure and the PRA 8 h after drug administration in normal- and high-renin patients (r = -0.623, p less than 0.05, n = 13), while no such relationship was observed in low-renin patients. The plasma aldosterone concentration (PAC) significantly decreased within 3 h, the lowest values occurring at 8 h after drug administration, and it returned to baseline levels within 24 h in all patients. No relationship was found between the reduction in mean blood pressure and changes in PAC after drug administration in either low-renin or normal- and high-renin hypertensives. The plasma bradykinin concentration (PBC) increased within 1 h, the highest values occurring at 3 h after drug administration, and returned to baseline levels within 24 h in low-renin hypertensives, while the PBC was significantly increased at 4 h and had not returned to baseline levels within 24 h in normal- and high-renin hypertensives. There was a significant correlation between percentage changes in mean blood pressure and those in PBC 90 min after drug administration in normal- and high-renin hypertensives (r = -0.556, p less than 0.05, n = 13), while no relationship was observed between them in low-renin hypertensives.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2991036 TI - Polyamine metabolism and virus replication. PMID- 2991037 TI - Applications of cellulases. PMID- 2991038 TI - On the mechanism of action of phenylalanine hydroxylase. PMID- 2991039 TI - Phenylalanine hydroxylase: metabolic aspects. PMID- 2991040 TI - Stimulation of oil biodegradation by using slow-release fertilizers. PMID- 2991041 TI - The mechanisms of LHRH agonist action in gonadal tissues. PMID- 2991042 TI - Electrofocusing fractionation of follicle stimulating hormone in pituitary cell culture extracts from male and female rats. AB - Three-day pituitary cell cultures from adult male and female rats were incubated for 4 h in the presence of 10 nM LHRH and the molecular heterogeneity of FSH was assessed in the media of LHRH-stimulated cells and in cell extracts from unstimulated cells using an electrofocusing technique. The pI distribution of FSH showed a high degree of similarity between cell media and cell extracts of each sex although differences were observed between sexes. Pituitary cell cultures from male rats were also incubated in the presence of 10(-8) M testosterone and 10(-8) M estradiol and the pI distribution of FSH from media after LHRH stimulation was determined. No significant differences in the pI profiles were observed. Incubation with charcoal-treated bovine follicular fluid (an inhibin source) resulted in a significant reduction in recovered FSH activity in the pH region 3.61-3.92 although this decrease did not markedly alter the pI profile of FSH. Close similarities were observed in the pI distribution of FSH of pituitary cell culture extracts and pituitary gland extracts from intact animals of both sexes, however, differences in pI distribution were noted in pituitary extracts in the male but not the female following gonadectomy. It is concluded that (1) stored FSH is released from the pituitary without major modification to its structure as assessed from its pI profile, (2) sex differences in the pI profile of FSH in pituitary extracts are retained in culture and following LHRH stimulated release, (3) the pI distribution of FSH is not affected by testosterone or estradiol and only minimally by inhibin in short-term cultures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991043 TI - Binding characteristics of epidermal growth factor receptors in male and female rat liver cell membrane preparations. AB - The binding characteristics of epidermal growth factor (EGF) receptors were studied in cell membrane preparations from adult male (n = 14) and female (n = 13) rat livers. Results indicate a significant lower (about 65%) number of EGF receptors in female preparations. The possibility that the decrease in EGF receptors was only a reflection of an excess free EGF in female preparations was ruled out by means of acid extraction, ultrafiltration, and measurement of EGF in the acid extracts. In view of the known role of EGF in cell differentiation, it may be important to recognize that the number of its receptors, at least in liver preparations, is markedly different between sexes. PMID- 2991044 TI - Identification of a monoclonal antibody which interacts with the parathyroid hormone receptor-adenylate cyclase system in murine bone. AB - We have produced monoclonal antibodies which bind specifically to mouse bone cells and then selected these monoclonal antibodies for their ability to inhibit parathyroid hormone (PTH) responses in mouse cranial bone treated with the (1-34) amino terminal peptide of bovine PTH [bPTH(1-34)]. One clone, designated 3-6, characterized as an IgM(kappa), significantly inhibited the accumulation of cAMP in response to bPTH(1-34) at concentrations of hormone between 10(-9) and 10(-7) M. This antibody was subsequently isolated by gel filtration and shown to bind to intact mouse calvariae, with saturation binding occurring at 3 micrograms/ml IgM. A maximal inhibition of approximately 70% of the cAMP accumulation produced in response to 2.5 X 10(-9) M (100 ng/ml) bPTH(1-34) was obtained with 7 micrograms/ml of the purified 3-6 IgM. At this concentration of 3-6 IgM, the half maximal dose of PTH for activation of cAMP accumulation was increased from 5 X 10(-9) M to 2 X 10(-8) M with no reduction in maximal levels of cAMP production. The utility of this antibody as an inhibitor was further tested by its ability to block the binding of an iodinated PTH analogue, 125I-[Nle8, Nle18, Tyr34]-bPTH(1 34) to mouse cranial bone. The 3-6 IgM at a concentration of 5 X 10(-8) M inhibited 70% of the specific binding of the 125I-labeled analogue. In the absence of parathyroid hormone, 2 X 10(-8) M 3-6 IgM produced a 4-fold increase in cAMP above basal levels, as compared to 40-fold maximal increases observed with PTH, indicating a partial PTH agonist activity of this antibody. When tested for effects on other hormones, 3-6 IgM did not inhibit cAMP accumulation produced in response to salmon calcitonin, epinephrine, prostaglandin E2 or cholera toxin. We propose that the 3-6 monoclonal IgM is specific for the PTH receptor or a component of the PTH receptor-adenylate cyclase system and that this or similar antibodies will serve as useful reagents for future molecular characterization of this receptor. PMID- 2991045 TI - The interaction between the TSH receptor and Graves' sera with TSH agonist or antagonist properties. AB - The A subunit of the TSH receptor was prepared by reduction of human thyroid membranes with dithiothreitol, and partially purified by gel filtration. The ability of Graves' sera to inhibit TSH binding to the TSH receptor and to stimulate cyclic AMP release from isolated thyroid cells was abolished by incubation with crude and partially purified preparations of the A subunit. PMID- 2991046 TI - Molecular cloning of a cDNA complementary to a UDP-glucose pyrophosphorylase mRNA of dictyostelium discoideum. AB - Uridine diphosphoglucose pyrophosphorylase (UTP: -alpha-D-glucose-1-phosphate uridyltransferase, EC 2.7.7.9) is an essential enzyme for normal development of Dictyostelium discoideum and its specific activity increases 3- to 10-fold by the later stages of development. Previous experiments have shown that additional forms of the enzyme appear concomitantly with this increase and that two uridine diphosphoglucose pyrophosphorylase (UDPGP) polypeptides are immunoprecipitated from the in vitro translation products of total cellular RNA at any stage of development (B. F. Fishel, R. E. Manrow and R. P. Dottin, 1982, Dev. Biol. 92, 175-187). Using an in vitro translation-immunoprecipitation assay of UDPGP mRNA, we show that an increase in the amount of translatable mRNA is correlated with the accumulation of enzyme during development. A cDNA bank was constructed from a mRNA population that had been enriched for UDPGP mRNA by size fractionation on sucrose gradients containing methylmercuric hydroxide (C. W. Schweinfest, R. W. Kwiatkowski, and R. P. Dottin, 1982, Proc. Natl. Acad. Sci. USA 79, 4997-5000). A 1.8-Kb cDNA complementary to a UDPGP mRNA was identified after screening the bank by hybridization selection and translation. Only the mRNA encoding the higher molecular weight in vitro translation product is hybrid selected by this cDNA. In hybrid-arrested translation experiments, the coding strand of this cDNA selectively inhibits the translation of only one of the two in vitro translation products. Therefore, there are two distinct UDPGP mRNAs. PMID- 2991047 TI - Developmental changes in expression of the Drosophila melanogaster epidermal growth factor receptor gene. AB - A Drosophila gene homologous to the human EGF receptor gene has recently been isolated and sequenced. Two transcripts, 7.6- and 7.1-kb long, encoded by this gene were identified in Drosophila melanogaster. The transcripts are present at low abundance in the maternal RNA stored in unfertilized eggs and in 2-hr-old embryos. The abundance of both transcripts increases sharply between 2 and 5 hr after egg deposition, it remains high throughout embryogenesis, and decreases again in the larval and pupal stages. In adult flies the two transcripts are expressed differentially. The 7.1-kb transcript is present during adulthood at the same level detected previously in pupal stage, but the abundance of the 7.6 kb transcript decreases substantially and it remains low during adulthood. PMID- 2991048 TI - Characteristics of the interaction of the glucagon receptor, cAMP, and insulin secretion in parent cells and clone 5F of a cultured rat insulinoma. AB - Rat insulinoma cells, which grow in culture and secrete insulin, were used to study the mechanism of stimulation of insulin release by glucagon. The parent cell line (RIN-m) and a clone that secretes high levels of insulin (5F) had been shown to possess specific receptors for glucagon. Glucagon (1 microM) stimulated a rapid increase in cyclic adenosine 3':5'-monophosphate (cAMP) that was followed by an increase in insulin secretion in both cell lines. The concentration of glucagon necessary for half-maximal stimulation of cAMP was 50 nM in parent and approximately 0.5 microM in 5F, whereas the concentration required to inhibit binding by 50% was 0.5 nM and 30 nM, respectively. In 5F, the dose-response relationships for cAMP and insulin secretion were superimposable. The glucagon effects on insulin secretion and cAMP did not require either glucose or amino acids in the incubation media. No refractoriness to glucagon stimulation of cAMP or insulin was noted. It may be concluded that there are significant differences between glucagon binding and glucagon responses in parent cells and clone 5F, there are glucagon receptors that are not coupled to adenylate cyclase, and cAMP mediates glucagon-stimulated insulin release. PMID- 2991050 TI - Hyperproinsulinemia in a family with a proposed defect in conversion is linked to the insulin gene. AB - Two previously described pedigrees with familial hyperproinsulinemia have elevated proinsulin conversion intermediates resulting from amino acid substitutions in the proinsulin molecule. In contrast, a third family with elevated levels of an apparently normal proinsulin molecule may have a defect in the converting process. To determine if the defect in this family lies in the insulin gene region, we used restriction fragment length polymorphisms adjacent to the insulin gene to examine cosegregation with hyperproinsulinemia. We demonstrate linkage of hyperproinsulinemia and the insulin gene in this family with a LOD score of 1.8, suggesting that the defect lies in or near the insulin gene. This method has wide applicability in determining whether hyperproinsulinemia or hyperinsulinemia is the result of defects at the insulin gene, and should permit the detection of new defects at or near this locus. PMID- 2991049 TI - Regulation of gluconeogenesis in hepatocytes from fasted alloxan-diabetic rats. AB - Hepatocytes from fasted, alloxan-diabetic rats were incubated in the absence of gluconeogenic substrates to deplete residual glycogen stores. Glucose production from lactate and pyruvate was enhanced in cells from diabetic rats relative to similarly treated hepatocytes from fasted, nondiabetic control rats. Gluconeogenesis from dihydroxyacetone, fructose, or glycerol was not increased but the formation of lactate plus pyruvate from dihydroxyacetone was decreased. The stimulation of gluconeogenesis by exogenous fatty acids was decreased by diabetes. The rates of gluconeogenesis in the presence of lactate plus pyruvate plus oleate were equal in hepatocytes from diabetic and control rats and indicate that the maximal rate of gluconeogenesis was not increased. With lactate plus pyruvate as substrates, stimulation of gluconeogenesis by norepinephrine or dibutyryl-cAMP was not altered by diabetes. The catecholamine stimulation of gluconeogenesis from glycerol also was unaffected. In contrast, diabetes decreased the maximal stimulation of gluconeogenesis from dihydroxyacetone by dibutyryl-cAMP, glucagon, or norepinephrine and this decrease was proportional to the decreased production of lactate plus pyruvate. The concentrations of glucagon or norepinephrine required for half-maximal stimulation were not altered by diabetes. Thus, the hormonal stimulation of gluconeogenesis from dihydroxyacetone is decreased by diabetes, probably because of decreased pyruvate kinase activity, but the interaction of glucagon and norepinephrine with hepatocytes and the subsequent stimulation of gluconeogenesis from physiologic substrates is not impaired. PMID- 2991051 TI - A prospective comparison of 'conventional' and high carbohydrate/high fibre/low fat diets in adults with established type 1 (insulin-dependent) diabetes. AB - Diabetic associations throughout the world are recommending high carbohydrate/high fibre/low fat diets for diabetic patients as a means of improving general health and metabolic control. We have previously shown that the efficacy of a 'conventional' low carbohydrate diet can be greatly increased by improving the method of dietary education. To test whether further improvement in metabolic control could be achieved by changing to a high carbohydrate/high fibre/low fat diet we have followed a group of 40 Type 1 (insulin dependent) diabetic adults for up to 19 months. They had all been prescribed a conventional low carbohydrate diet taught by one of three methods (dietician only, practical lunchtime demonstration or videotape). Those taught by dietician only were left as a control group for follow-up (group A). The others were randomized either to continue on their present diet (group B) or to change to a high carbohydrate/high fibre/low fat diet (group C). At final assessment those in groups B and C were more knowledgeable, compliant and better controlled than group A. Those in group C achieved a change from 38% carbohydrate/43% fat/20 g fibre daily to 45% carbohydrate/34% fat/32 g fibre daily. However, metabolic control in group C deteriorated while taking the high carbohydrate/high fibre/low fat diet (haemoglobin A1 went from 9.4 +/- 0.5% to 11.2 +/- 0.5%, p less than 0.01, over 4 months). None of the patients in group C changed their eating habits or those of their families nearly as much as was recommended.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991052 TI - Type 1 (insulin-dependent) diabetes and a highly variable locus close to the insulin gene on chromosome 11. AB - A polymorphic DNA sequence in the 5'-flanking region of the human insulin gene was studied in relation to Type 1 (insulin-dependent) diabetes. In 141 Caucasoid subjects analysed by Southern blot hybridisation techniques, two major DNA insertions were observed: a Class 1 allele or a Class 3 allele. The Class 2 allele was not observed in this group of subjects. Genotype frequencies in a control population (n = 88) were: homozygous 1/1, 42%; heterozygous 1/3, 50%; and homozygous 3/3, 8%. Corresponding genotype frequencies in 53 Type 1 diabetic patients were 79%, 21% and 0%, respectively (p less than 0.0005 from chi 2 test). This confirms prevalence data reported by Bell et al. [16]. There appeared to be no coinheritance with HLA-DR3/DR4 related antigens, nor with autoimmune features. Analysis of 17 Type 1 diabetic pedigrees including 34 diabetic and 69 non diabetic subjects did not demonstrate genetic linkage of these DNA inserts with diabetes, using an autosomal recessive, single locus model of inheritance. PMID- 2991053 TI - Insulin-sensitive phosphodiesterase and insulin receptor binding in fat cells from spontaneously obese rats. AB - The effects of insulin on insulin-sensitive phosphodiesterase were investigated in fat cells from rats aged 4, 8 and 16 weeks. The enzyme activities in rats aged 4 and 8 weeks higher at 0.1-30 nmol/l insulin concentrations than in rats aged 16 weeks, and half-maximum stimulations were obtained at 0.08 nmol/l in rats aged 4 weeks, at 0.15 nmol/l in rats aged 8 weeks and at 0.22 nmol/l in rats aged 16 weeks. Specific binding of insulin in fat cells from rats aged 4, 8 and 16 weeks was 3.3%, 5.0% and 11.6%/2 X 10(5) cells, respectively. Scatchard analysis indicated that increased insulin binding in fat cells from rats aged 16 weeks was due mainly to an increase of binding affinity. These results suggest that impairment of the phosphodiesterase activation system in fat cells from spontaneously obese rats is predominantly due to post-receptor defects. PMID- 2991054 TI - Untoward effects of pharmacological doses of insulin in early chick embryos: through which receptor are they mediated? AB - The teratogenic effect of insulin in early vertebrate embryos is controversial and the mechanisms involved are unknown. We studied the effects of pharmacological doses of insulin in chick embryos during the period of differentiation. We compared the effects of insulin with two proinsulins, desoctapeptide-insulin and multiplication-stimulating activity, peptides that have little insulin-like metabolic activity while they have significant growth effects. Chick embryos at 46 h of development were injected with the different peptides. At 96 h the mortality and abnormal growth elicited by the peptides were dose-dependent. Considering the indices of lethality (LD50) and affected embryos (ED50) as 100% for insulin, proinsulin was 59-66% as potent as insulin, desoctapeptide-insulin 2-6% and multiplication-stimulating activity 176-204%. In the surviving embryos, insulin (5 micrograms, decreased DNA, RNA and protein content by 49%, 40% and 48% respectively compared with controls. The effects of insulin were not corrected by simultaneous glucose injections. These data suggest that insulin, at pharmacological doses, interferes with embryo development through a non-metabolic pathway, probably via a growth-type receptor. PMID- 2991055 TI - Constancy of DNA organization of polymorphic and nonpolymorphic genes during development in Xenopus. AB - A study was undertaken to test for the occurrence of DNA rearrangements or amplifications during embryonic development in Xenopus laevis. DNA isolated from testes and liver was digested with four restriction enzymes, separated on agarose gels, transferred to nitrocellulose, and hybridized with over 50 cloned cDNA probes generated from embryonic poly (A)+ RNA. No qualitative or quantitative differences were detectable in the DNA hybridization patterns of testes and liver DNA, suggesting that, at least during liver development, selective amplifications or rearrangements occur rarely if at all. In the course of this investigation a wide range of restriction-site polymorphisms for different genes was observed. While some genes showed little polymorphism among different animals, several genes showed considerable polymorphism, involving changes in several restriction enzyme sites. These complex polymorphisms could be the result of gene rearrangements that occur occasionally during the course of sexual reproduction rather than during development. PMID- 2991056 TI - A teratocarcinoma-derived endoderm stem cell line (1H5) that can differentiate into extra-embryonic endoderm cell types. AB - We investigated the ability of the teratocarcinoma-derived, epithelial-type cell line 1H5 to differentiate into either of the two pathways to primary endoderm, and tested the hypothesis that 1H5 represents a state similar to primitive endoderm in the late 4th-day blastocyst. Like other endodermal cell types, 1H5 cells mixed with embryonal-carcinoma cells sort out into "embryoid bodies" or structures that resemble 4th-day mouse embryos. The epithelial line conforms morphologically and biochemically to the few known characteristics typical of primitive endoderm. The present study demonstrates that the formation in vitro of overt visceral endoderm is readily achieved. The spontaneous arrangement of the cells into a cystic form is followed by the appearance of several markers of visceral endoderm, most notably alphafetoprotein, which is detected when 1H5 cells are cultured either in the presence of retinoic acid or when the cells interact with embryonal-carcinoma cells in a specific spatial arrangement after sorting out. However, some less specific properties of visceral endoderm are not expressed. Although 1H5 differentiates histologically into parietal-like endoderm in the tumor form, parietal cells cannot yet be identified with certainty in vitro because of the paucity of parietal-specific markers. The 1H5 cell line could provide a useful system for studying the characteristics and mechanisms underlying visceral-endoderm differentiation in vitro, since it has the distinct advantage that homogeneous cultures are produced, in contrast to other teratocarcinoma cell lines such as F9 which differentiate into a mixture of cell types. PMID- 2991057 TI - Induction of undifferentiated brain tumors in rats by a human polyomavirus (JC virus). AB - Newborn SD rats were inoculated intracranially with JC virus (Tokyo-1), a human polyomavirus, isolated from the autopsied brain of a patient with progressive multifocal leukoencephalopathy. Twenty-one to 61 weeks later, 20 of 27 rats developed tumors in the cerebrum, but not in the cerebellum. The undifferentiated neuroectodermal nature of the tumors was histologically, immunohistochemically and ultrastructurally confirmed. PMID- 2991058 TI - Quantitative analysis of AIDS-related virus-carrying cells by plaque-forming assay using an HTLV-I-positive MT-4 cell line. AB - Quantitation of acquired immune deficiency syndrome (AIDS)-related virus-bearing cells was analyzed by a newly developed plaque-forming assay. A chemically adherent HTLV-I-positive MT-4 cell monolayer was used as the cytolytic responder cells to the AIDS-virus infection. When AIDS-virus-infected cells (Molt-4/HTLV III, H9/HTLV-III, and CEM/LAV) were seeded with MT-4 cells, plaques were observed corresponding to the number of HTLV-III-positive cells plated. Plaque formation was inhibited by the addition of anti-HTLV-III-positive serum (neutralizing antibody) to the agarose medium. These data suggest that this plaque-forming assay technique should be useful for the quantitative analysis of AIDS-virus bearing cells. PMID- 2991059 TI - Two distinctive transforming DNA regions on the canine adenovirus type 1 genome. AB - By transfecting cloned DNA fragments of canine adenovirus type 1, a novel transforming region of mammalian adenovirus was demonstrated inside the viral genome. Besides the left-end SmaI-D fragment (left 17.2%), an internally located SmaI-E fragment (17.2-28.7%) of canine adenovirus type 1 was found to transform primary rat kidney cells. PMID- 2991060 TI - Mother-to-child transmission of human T-cell leukemia virus type-I. AB - By screening sera obtained from 5015 pregnant women under the care of 9 gynecology/obstetrics departments of hospitals or clinics in Nagasaki City and its surrounding areas, 187 were found to be positive for antibody against adult T cell leukemia-associated antigen (ATLA). The prevalence of seropositive pregnant women was consistent with that of blood donors in the age group of 16 to 29 years, taken as controls. Essentially all cord blood samples (113/115) of babies born from these seropositive mothers were positive for anti-ATLA of IgG class. These IgG antibodies in the babies diminished rapidly after delivery, and were detectable only in 3 cases at 2, 3, and 5 months of ages out of 38 babies up to 21 months. None of 115 cord bloods so far tested was positive either for anti ATLA of IgM class or for ATLA-bearing lymphocytes after short-term cultures of the rosette-forming T-lymphocytes. Two of the babies born from 38 seropositive mothers were found seroconverted between the ages of 12 and 19 months. Three out of 20 elder siblings were found seropositive at ages of 3 to 6 years. The seropositive rate in these siblings was significantly higher than that of controls. Moreover, 12 out of 13 mothers traced back from seropositive students were found seropositive. These data indicate strongly that HTLV-I is transmitted from seropositive mothers to their children as a major pathway. PMID- 2991061 TI - Oral transmission of human T-cell leukemia virus type-I into a common marmoset (Callithrix jacchus) as an experimental model for milk-borne transmission. AB - To obtain experimental support for possible milk-borne infection of human T-cell leukemia virus type-I (HTLV-I), short-term-cultured viral antigen-positive lymphocytes obtained from peripheral blood of adult T-cell leukemia lymphoma complex (ATLL) patients were inoculated into the oral cavity of two adult common marmosets (Callithrix jacchus) in amounts comparable to those of HTLV-I-carrying cells fed to a baby in the milk of seropositive mothers. One of the animals seroconverted 2.5 months after the first inoculation. Cultures of peripheral blood mononuclear leukocytes of the marmoset revealed HTLV-I antigen expression in cells, indicating the establishment of oral infection of HTLV-I in an adult marmoset. The cell number deduced to be responsible for the infection was 5.6 X 10(7) cells (used in the first 2 inoculations). The results suggest that the concept of milk-borne infection of HTLV-I from a seropositive mother to her child is plausible. PMID- 2991062 TI - Increase in DNA polymerase alpha in nuclei of chicken kidney cells after avian adenovirus infection. AB - DNA polymerase activity which sedimented at about 7s in the presence of 0.5M KC1 increased in cultured kidney cells after infection with a chicken embryo lethal orphan virus, an avian adenovirus. Elevated levels of antigenic material to a monoclonal antibody specific for the large subunit of chicken DNA polymerase alpha were also detected in nuclei of virus-infected cells by an immunofluorescence method. Since the elevations of DNA polymerase activity and of nuclear immunofluorescence was not inhibited with 1-beta-D arabinofuranosylcytosine, the early viral gene(s) seems to be responsible for the induction of the synthesis and nuclear accumulation of DNA polymerase alpha. PMID- 2991063 TI - Papillomavirus infection among Japanese: an immunohistochemical study for papillomavirus genus-specific antigen in human surface epithelial lesions. AB - In order to clarify the relationship between human papillomavirus (HPV) and a variety of surface epithelial lesions, the presence of papillomavirus genus specific common structural antigen (pgs-antigen) was immunohistochemically investigated in 256 cases of various tumors and tumorous lesions. The pgs-antigen was demonstrated in cases of verruca vulgaris (11/23 cases), condyloma acuminatum (13/26), adult laryngeal papilloma (3/12) and bowenoid papulosis (2/2). No pgs antigen was observed in ordinary Bowen's disease and other hyperkeratotic skin lesions, such as keratoacanthoma and seborrheic keratosis. In uterine cervical lesions, about 15% of cervical dysplasia, most of which later developed into carcinoma in situ, contained pgs-antigen-positive koilocytotic cells. These results suggest that HPV infection is frequently present in human hyperplastic and atypical surface epithelial lesions of Japanese patients and might indicate possible association with neoplastic transformation, especially in the cervix and skin. PMID- 2991064 TI - Prognostic value of estrogen and prolactin receptor analysis in human breast cancer. AB - Hormone receptors for estrogen (ER), progesterone (PGR) and prolactin (PRL-R) were measured in primary breast cancer tissues obtained from 214 patients at radical mastectomy. The patients were followed up at intervals of one to three months to examine the relationship between hormone receptor status and prognosis. ER status was related to PGR status but not to PRL-R status. PRL-R was more frequently detectable in pathologic stage 3 than in stage 1 & 2 (22% vs. 10%, P less than 0.05) whereas ER and PGR were not different between stage 1 & 2 and stage 3 groups. The influence of receptor status on prognosis was analyzed in 164 patients of stage 1 & 2 by means of the actuarial life table technique. The recurrence-free interval was not related to ER, PGR or PRL-R status. The ER positive group was associated with significantly prolonged survival compared with that of ER-negative patients at 42 to 51 months after surgery. PRL-R positive patients had a significantly worse survival than the PRL-R negative group (P less than 0.05). These findings indicate that receptor status may provide useful prognostic information in patients with early breast cancer and that the lactogenic hormone may play an unfavorable role in relation to the prognosis of human breast cancer. PMID- 2991065 TI - [Acute ulcerative colitis and cytomegalovirus infection in a nonimmunosuppressed patient: delayed appearance of specific antibodies]. PMID- 2991066 TI - Human prolyl hydroxylase. Purification, radioimmunoassay and clinical studies in liver diseases. AB - Prolyl hydroxylase was purified from human placentae, specific antiserum against it was prepared, and a new radioimmunoassay system employing 125I-labelled enzyme preparation was established. The molecular weight of the placental enzyme was shown to be 320,000 by gel filtration. SDS-polyacrylamide gel electrophoresis showed two bands of unequal intensity having molecular weights of 60,000 and 130,000. Their amino acid compositions were identical to each other, suggesting the polypeptide with a molecular weight of 130,000 might be a dimer of the polypeptide with a molecular weight of 60,000. The new radioimmunoassay established had a sensitivity of the order of 10 ng/ml, indicating it was more sensitive than previous radioimmunoassay employing 3H-labelling method. Clinical studies on patients with liver diseases disclosed that the concentrations of serum immunoreactive prolyl hydroxylase were elevated both in cases of hepatocellular damage and in cases of cholestasis. In cases of hepatocellular damage the enzyme behaved like cytoplasmic enzymes such as glutamic oxaloacetic transaminase, glutamic pyruvic transaminase and lactic dehydrogenase, but in cases of cholestasis it resembled biliary enzymes such as alkaline phosphatase and gamma-glutamyl transpeptidase. This result might be associated with the peculiar location of the enzyme within the cell, in the membrane of rough endoplasmic reticulum. PMID- 2991068 TI - Effects of vegetable diets on nitrogen metabolism in cirrhotic subjects. AB - This study compared the effect of a vegetable diet with an animal protein diet on various aspects of nitrogen metabolism to identify what components of the vegetable diet might be causing beneficial therapeutic effects in hepatic encephalopathy. Vegetable diets contained 4.5-fold greater amounts of fiber (56 +/- 3 g/day) and reduced amounts of methionine, tyrosine, and tryptophan. In 6 stable cirrhotic subjects without encephalopathy, vegetable diets caused a significant reduction in the urea production rate from 106 +/- 5 to 89 +/- 5 mg X kg-1 X 24 h-1 of urea nitrogen. This was mainly accounted for by a fall in urinary urea output. Vegetable diets also caused a fall in total urinary nitrogen, which was accounted for by the fall in urea nitrogen, and a comparable increase in fecal nitrogen from 12 +/- 2 to 28 +/- 5 mg X kg-1 X 24 h-1. The fecal bacterial fraction contained 63% of the increase in stool nitrogen. Most plasma amino acids, including methionine, tyrosine, phenylalanine, as well as total and free tryptophan, were unchanged. The effect of vegetable diets on nitrogen metabolism can be mainly accounted for by the increased intake of dietary fiber and increased incorporation and elimination of nitrogen in fecal bacteria. PMID- 2991067 TI - An epidemic of hepatitis A related to ingestion of raw oysters. AB - An epidemic of hepatitis A occurred around Hondo City, Kumamoto Prefecture in Japan during the first six months of 1982. Clinical, immunological and epidemiological studies were carried out in 225 cases. Cases were distributed over a relatively wide area, and in small numbers of young children and school children. More than half of the patients were in their twenties or thirties. The clinical course was generally favorable with rapid resolution. No episode lasted more than six months. There was only one fatality in a cure which was a carrier of HBs antigen with liver cirrhosis. Titers of IgM anti-HAV measured by radioimmunoassay (RIA) or enzyme immunoassay (EIA) reached a peak during the second week after onset, followed by a gradual decrease. Conversion to negative results was never experienced within two months. We found a good correlation between RIA and EIA in terms of detecting IgM anti-HAV. The route of infection was thought to be fecal-oral in nature, with ingestion of raw oysters the major etiologic factor. PMID- 2991069 TI - Dynamics of collagen accumulation and polymorphism in murine Schistosoma japonicum. AB - The dynamics of hepatic collagen biosynthesis and degradation were studied in mice infected with Schistosoma japonicum. Hepatic fibrosis, the major clinical manifestation of disease, increased during acute infection (0-15 wk). The majority of proline incorporation into hydroxyproline, which was reflective of collagen synthesis, was found within hepatic egg granulomas. As the disease became chronic (20-30 wk) there was a decrease in collagen synthesis and a maintenance of total collagenolytic activity, which resulted in decreased accumulations of both total hepatic and granuloma-associated collagen. In addition to these quantitative decreases in extracellular collagen there was a qualitative change in the type of hepatic collagen synthesized. Early in infection, type I collagen was the predominant biosynthetic product, whereas late in infection type III collagen became the dominant isotype. A similar switch was seen in the substrate specificity of the constitutive collagenolytic activity, with decreasing type I activity and increasing type III activity as the disease progressed from acute (10 wk) to chronic (30 wk). These changes in the quantity and makeup of extracellular collagen may lead to amelioration of disease and potential reversibility of fibrosis. PMID- 2991070 TI - Somatosensory evoked potentials in hepatic encephalopathy. AB - Median nerve somatosensory evoked potentials were recorded from 33 patients with various degrees of hepatic failure and from 10 age-matched controls. Within 20 ms poststimulation, one negative peak (N13) could be recorded from the middle of the back of the patient's neck at the C2 vertebral level. Within 150 ms, three negative and three positive peaks, sequentially designated as N1, P1, N2, P2, N3, and P3, could be recorded from the scalp over the contralateral sensory cortex. There was a progressive prolongation of peaks and interpeak latencies correlating with the severity of hepatic encephalopathy. In 10 patients with hepatic failure but no clinical evidence of hepatic encephalopathy, latencies of peak N3 and P3 were delayed and N1-N3 interpeak latencies were prolonged. Thirteen patients with grade 1 or 2 hepatic encephalopathy showed further delayed latencies of peaks P2, N3, and P3, further prolonged N1-N3, N1-P2 interpeak latencies, and distortion of waveforms. Peaks N2, P2, N3, and P3 were further delayed, and even disappeared in 10 patients with grade 3 or 4 hepatic encephalopathy. However, central conduction time (N13-N1 interpeak latency) was not prolonged in all stages of hepatic failure. In addition, serial somatosensory evoked potential studies correlated well with the clinical course. The present data suggest that somatosensory evoked potential recording is a reliable objective method in the early assessment and monitoring of hepatic encephalopathy. PMID- 2991071 TI - Neutralizing antibody to hepatitis A virus in immune serum globulin and in the sera of human recipients of immune serum globulin. AB - We determined the titer of neutralizing antibody to hepatitis A virus in five lots of immune serum globulin and in the sera of human recipients of immune globulin using a new and sensitive procedure, the radioimmunofocus inhibition assay. The neutralizing antibody titer of four immune globulin lots ranged from 1:170,000 to 1:406,000, and was approximately 100-fold the antibody titer determined by radioimmunoassay. The neutralizing antibody titer of a Bureau of Biologics reference immunoglobulin was 1:794,000. Sera collected from 18 healthy men who had undergone prophylaxis with immune globulin (0.02 ml/kg body wt) were negative when tested by radioimmunoassay. However, 2 of 18 sera collected before administration of immune globulin and sera from all 18 men collected 3 and 55 days later contained detectable neutralizing antibody. The measurement of neutralizing antibody should prove useful for standardizing passive immunoprophylaxis against hepatitis A as well as for evaluating the potential efficacy of new hepatitis A virus vaccines. PMID- 2991072 TI - Androgen receptors in hepatocellular carcinoma and surrounding parenchyma. AB - The androgen receptor content of normal human liver, hepatocellular carcinoma, and surrounding liver tissue was determined in patients with chronic liver disease. Androgen receptor was detected in all six normal livers obtained from 4 men and 2 women. The androgen receptor content in these 6 individuals ranged from 5.0 to 10.2 fmol/mg protein (Kd 10.6-31.8 X 10(-10) M). The livers from 2 patients with chronic active hepatitis and from 10 cirrhotic patients with hepatocellular carcinoma had detectable amounts of androgen receptor ranging from 2.0 to 14.8 fmol/mg protein (Kd 4.0-30.9 X 10(-10) M). Androgen receptor was found in the cytosol of 14 of 19 men with hepatocellular carcinoma. The titer ranged from 3.7 to 45.4 fmol/mg protein (Kd 3.2-21.4 X 10(-10) M). Hepatocellular carcinoma had a significantly higher concentration of androgen receptor than did the surrounding cirrhotic liver tissue. In 2 men and 1 woman, androgen receptor was detected in the cirrhotic liver but not in the tumor. In the remaining 3 men, both tumor and cirrhotic liver were negative for androgen receptor. PMID- 2991073 TI - Behavioral and neurochemical aspects of prostaglandins in brain function. PMID- 2991074 TI - Activatory effect of two cardioglycosides on Cavia cobaya kidney Na+/K+-ATPase activity. AB - Ouabain and K-strophanthoside promote an enhancement of Na+/K+-ATPase activity in a range of cardioglycoside concentrations from 100 nM to 100 pM, with a maximum (+30%) between 10 and 4 nM. Binding experiments with [3H]ouabain show upward curved Scatchard plots and evidence two intrinsic affinity constants for the ligand: (a) High-affinity constant: 350 nM (microsomes) and 15 nM (purified enzyme). (b) Low-affinity constant: 2100 nM (microsomes) and 890 nM (purified enzyme). The reaction velocity trend indicates that at ouabain concentrations higher than 20 nM but lower than the minimal inhibiting level, the enhanced reaction velocity is tending towards the control values. PMID- 2991076 TI - Adrenocortical responses to novel stressors in acutely or repeatedly starved chickens. AB - The effect of starvation on the corticosterone responses of immature cockerels to acute, novel stress has been determined. The marked corticosterone responses of fed birds to either horizontal treadmill exercise (0.04 km/hr) or intravenous adrenocorticotrophic hormone (ACTH) administration (P less than 0.001 in both cases) were reduced by starvation (P less than 0.01 and P less than 0.001, respectively). This reduction did not appear to be due to either feedback inhibition of corticosterone on the hypothalamus or pituitary, or to reduced adrenal responsiveness to endogenous ACTH. Starvation significantly elevated the basal level of circulating corticosterone (P less than 0.001), but the magnitude of this elevation and the level of corticosterone attained were less (P less than 0.05) in birds that were accustomed to starvation. This habituation of adrenocortical activity may be due to reduced pituitary ACTH secretion, and was specific in that the corticosterone responses to novel stressors were unaffected. PMID- 2991075 TI - Transitory corticosterone responses of ducks (Anas platyrhynchos) to exercise. AB - Horizontal treadmill exercise induced a marked (P less than 0.001) but transitory increase in the level of circulating corticosterone in the plasma of adult male ducks. The decline in corticosterone concentration during exercise is unlikely to be due to a depletion of adrenocortical stores since a marked (P less than 0.001) corticosterone response to adrenocorticotrophin (ACTH) administration was observed immediately after exercise, and was of similar magnitude to that induced in nonexercised controls. The corticosterone response to repeated exercise is also transitory, habituating completely within 28 days of the start of daily training. The corticosterone response (P less than 0.001) to ACTH challenge is not, however, diminished by training. These results indicate that the habituation of the corticosterone response to either acute or repeated exercise is due to a reduction in endogenous ACTH secretion. A similar mechanism appears to be responsible for the habituation of the corticosterone response to handling and confinement in nonexercised control birds. A marked (P less than 0.001) increase in the level of circulating corticosterone was, however, elicited when these birds were exercised, indicating that despite adaptation to handling and confinement the birds remained responsive to the novel stressor of exercise. The corticosterone response of these birds was, however, less than that in untrained birds indicating that the corticosterone response in untrained birds is due to both workload and the stress of handling and confinement. PMID- 2991077 TI - Factors affecting the secretory dynamics of 1 alpha-hydroxycorticosterone in the dogfish, Scyliorhinus canicula. AB - The secretory dynamics (plasma concentrations, metabolic clearance rates (MCR), and blood production rates (BPR] of 1 alpha-hydroxycorticosterone (1 alpha OH-B) were examined in Scyliorhinus canicula under a number of experimental conditions. Both hypophysectomy and injections of dexamethasone significantly reduced the plasma concentrations of 1 alpha OH-B, although significant, measurable amounts remained. In hypophysectomized animals MCR increased and BPR decreased. Intravenous administration of Squalus and mammalian adrenocorticotrophic hormone (ACTH) preparations, homologous renal extracts, and mammalian angiotensin II all increased plasma concentrations of 1 alpha OH-B; apart from Squalus ACTH these materials increased BPR of 1 alpha OH-B and both ACTH preparations reduced the MCR. In none of the experimental groups were there major changes in plasma osmolality or in sodium, potassium, magnesium, chloride, calcium, or urea concentrations. It is concluded that the secretory patterns of 1 alpha OH-B are significantly influenced by renal materials in addition to hypophysial ACTH. PMID- 2991078 TI - Identification by immunofluorescence of ACTH-producing cells in the pituitary gland of the tree frog Hyla arborea. AB - The indirect immunofluorescence procedure was used to localize ACTH-producing cells in the pituitary distal lobe from Hyla arborea adult specimens; the antiserum employed was rabbit anti-synthetic ACTH (1-24) conjugated with BSA. Immunohistochemical staining was suppressed by solid-phase absorption of the anti ACTH with the specific antigen. In the distal lobe one cell population is immunoreactive; these cells are predominantly localized in the anterior half, both ventrally and dorsally; they exclusively correspond to type-3 basophils. In the intermediate lobe the total cell population is immunoreactive. Parallel inspections carried out on the pituitary of Xenopus laevis specimens gave results fully consistent with those reported by other authors. PMID- 2991079 TI - Elevation of plasma glucose, alanine, and urea levels by mammalian ACTH in the American bullfrog (Rana catesbeiana). AB - The effects of a single infusion of mammalian ACTH on plasma glucose, alanine, urea, and lactate were determined in the American bullfrog (Rana catesbeiana). The ACTH (10 U/250 g body wt) was administered, and serial blood samples were collected via a nonocclusive cannula chronically placed in the right truncus arteriosus. Plasma metabolite levels were estimated by standard enzymatic techniques. The plasma metabolites declined following the surgery to levels that were relatively stable by postoperative Day 2. The levels did not vary significantly for the remainder of the 3- or 4-day pretreatment period and in the control bullfrogs during the 48-hr experiments. Plasma glucose levels were essentially unchanged from the time-zero levels at 6 hr following ACTH infusion. Plasma glucose levels subsequently increased to levels that were approximately 24% greater than the control levels by 24 hr and then declined to near control levels by 48 hr. Plasma alanine increased to levels that were approximately 60% greater than the control levels by 12 hr after ACTH treatment and returned to essentially the time-zero levels by 24 hr. Plasma urea rose to levels that were approximately 110% greater than the control levels by 45 min after ACTH infusion, but urea returned to essentially the time-zero levels by 1.5 through 3 hr. Plasma urea increased again to levels that were approximately 90% greater than the control levels by 6 hr and returned to essentially the initial levels by 24 hr. Plasma lactate levels were not significantly influenced by ACTH treatment. The results suggest that a function of the bullfrog hypothalamic-pituitary adrenocortical axis is to regulate gluconeogenesis from alanine, and probably other glucogenic amino acids. PMID- 2991080 TI - Thyroid function in adult rough-skinned newts (Taricha granulosa): effects of hypophysectomy and hormone replacement. AB - This study examines the regulation of thyroid activity in adult rough-skinned newts, Taricha granulosa. Aquatic, breeding-condition newts were hypophysectomized and were injected for 23 days with the mammalian adenohypophysial hormones: ACTH, PRL, and GH. During a fourth week of treatment, these newts also received TSH. Plasma T4 concentrations were lower in hypophysectomized newts than in sham-hypophysectomized newts 23 days after surgery. Both ACTH and PRL increased T4 titers above these in saline-injected control newts; GH alone had no effect. TSH effectively increased the plasma T4 concentrations in newts from all five groups. GH, and possibly PRL, depressed the response to TSH. Plasma T4 concentration was positively correlated with body fluid loss, suggesting that hormone concentration/dilution effects may occur generally in animals such as these that undergo substantial seasonal changes in degree of hydration. PMID- 2991082 TI - Genetic and physical studies of a portion of the white locus participating in transcriptional regulation and in synapsis-dependent interactions in Drosophila adult tissues. AB - We have identified and sequenced the portion of the white locus affected by an idiosyncratic set of white mutant alleles (the wsp alleles). The affected white locus portion (wsp region) extends from ca. 590 base pairs (bp) to ca. 1270 bp 5' to the apparent start site for the major white transcription unit. Based on the properties of these mutant alleles, we infer the existence of two distinct cis acting regulatory elements in the wsp region and a third element mapping 3' to this region (3' to position ca. -670). Our analysis allows us to define the apparent position of one of the two wsp region elements with substantial precision. Examination of the DNA sequences in this region suggests that it is functionally similar to the enhancers identified in vertebrates. This same element participates in synapsis-dependent genetic interactions, suggesting a largely unexpected relationship between enhancer-like, cis-acting genetic elements and the genetic elements responsible for the synapsis-dependent genetic interactions in trans revealed by the existence of transvection effects. Our results further suggest that a presumptive regulatory locus (suppressor-of-white spotted) regulates white transcription in adult tissues and is not involved in regulating white expression in larvae. We discuss the regulation of white expression in light of our studies. We also demonstrate unusual structures for an X-ray-induced deletion and a spontaneous deletion. PMID- 2991083 TI - A model of duplicative transposition and gene conversion for repetitive DNA families. AB - A model of duplicative transposition and gene conversion for the evolution of repetitive DNA families was studied. In this model, transposition and conversion (both unbiased) are assumed to occur both within and between the genomes in a diploid cell, and any degree of linkage intensity is incorporated. The transition equations for allelic and nonallelic identity coefficients have been formulated by using the previous results. The results are widely applicable to many repetitive sequences, from dispersed families like transposons to tightly linked multigene families. It has been shown through extensive numerical studies on equilibrium properties that duplicative transposition and gene conversion have very similar effects on nonallelic identity coefficients, but that allelism and allelic identity are greatly influenced by the relative rates of occurrence of the two processes. PMID- 2991081 TI - Resolution of dicentric chromosomes by Ty-mediated recombination in yeast. AB - We have integrated a plasmid containing a yeast centromere, CEN5, into the HIS4 region of chromosome III by transformation. Of the three transformant colonies examined, none contained a dicentric chromosome, but all contained a rearranged chromosome III. In one transformant, rearrangement occurred by homologous recombination between two Ty elements; one on the left arm and the other on the right arm of chromosome III. This event produced a ring chromosome (ring chromosome III) of about 60 kb consisting of CEN3 and all other sequences between the two Ty elements. In addition, a linear chromosome (chromosome IIIA) consisting of sequences distal to the two Ty elements including CEN5, but lacking 60 kb of sequences from the centromeric region, was produced. Two other transformants also contain a similarly altered linear chromosome III as well as an apparently normal copy of chromosome III. These results suggest that dicentric chromosomes cannot be maintained in yeast and that dicentric structures must be resolved for the cell to survive.--The meiotic segregation properties of ring chromosome III and linear chromosome IIIA were examined in diploid cells which also contained a normal chromosome III. Chromosome IIIA and normal chromosome III disjoined normally, indicating that homology or parallel location of the centromeric regions of these chromosomes are not essential for proper meiotic segregation. In contrast, the 60-kb ring chromosome III, which is homologous to the centromeric region of the normal chromosome III, did not appear to pair with fidelity with chromosome III. PMID- 2991084 TI - [Modular structure of the genes of phages-transposons of Pseudomonas aeruginosa]. AB - The basic criterion to confirm the recombinational origin of bacteriophages belonging to the same phage family is revealing several different combinations of differentiated segments in phage genomes which determine specific functions (modules). The results of phage-to-phage comparison of several regions in genomes of closely related transposable phages of Pseudomonas aeruginosa D3112, B39, PH2, PH51, PH93, PH132 have supported the modular hypothesis for this group of phages. PMID- 2991085 TI - [Characteristics of phages-transposons of Pseudomonas aeruginosa belonging to 2 groups distinguished by DNA-DNA homology]. AB - 14 new transposable phages (TP) were isolated from approx. 200 clinical isolates of Pseudomonas aeruginosa. The frequent occurrence of TP of P. aeruginosa has been confirmed. There are at least two different groups of TP, namely, the group of D3112 and that of B3. The distinctive features of phages belonging to the groups are as follows: 1) low level of DNA-DNA homology (less than 10%), the whole region of homology in phage genomes of different groups being located on right genome end (29-38 kb); only one of phages of the B3 group shows an additional homology with D3112 DNA outside the above mentioned region; 2) a variable DNA is observed on the left end of the B3 group phage genomes and no such DNA is revealed on the left end of genomes of the D3112 group phages; 3) all phages of the B3 group have specific type of interaction with RPL11 plasmid, which distinguish them from phages of the D3112 group; 4) phages belonging to the two groups differ greatly in their growth in cells harbouring pMG7 plasmid which mediates production of PaeR7 endonuclease and in the number of DNA sites sensitive to SalGI, PstI, BglII endonucleases. Since some of the B3 group phage genomes possess BamH1 sites, resistance to this enzyme cannot be regarded as a general characteristics for all TP of P. aeruginosa, as it was earlier proposed. Some aspects of modular hypothesis of bacteriophage evolution concerning, in particular, the ways of module formation are discussed. PMID- 2991086 TI - [Physical and genetic organization of the plasmid R15]. AB - The conjugative IncN plasmid R15 (SmrSurHgr, 62.3 kb) is cleaved by the hexanucleotide-specific endonucleases BglII, HindIII, EcoRI, BamHI, SmaI, SalI, PstI and XhoI into 9, 9, 6, 5, 4, 4, 4 and 2 fragments, respectively. The restriction sites were located on the physical map of the R15 genome. Distribution of the cleavage sites is strongly asymmetric. 28 of 32 sites for BamHI, EcoRI, HindIII, SalI, SmaI and PstI were located close to or within the sequences of transposable elements Tn2353 and Tn2354. According to the results of analysis of R15::Tn1756 deletion derivatives and recombinant plasmids harboring fragments of R15, the genetic determinants for resistance to Sm, Su and Hg were mapped, as well as the regions necessary for EcoRII restriction--modification and for plasmid replication and conjugation. The features of physical and genetic structures of R15 and other IncN plasmids are discussed. PMID- 2991087 TI - [Assessment of the maximum permissible concentration of quartz dust in the air by its experimentally established maximum permissible content in the lungs]. PMID- 2991088 TI - [Importance of the method of stimulation electromyography for the early diagnosis of peripheral neuropathy from lead exposure]. PMID- 2991089 TI - Faecal mucus degrading glycosidases in ulcerative colitis and Crohn's disease. AB - Because the normal faecal flora includes bacteria which can produce mucus digesting glycosidases, it follows that increased digestion of colonic mucus by these bacterial enzymes could be important in the pathogenesis of ulcerative colitis. Faecal activities of potential mucus-degrading glycosidases have therefore been assayed in samples from patients with inflammatory bowel disease and normal controls. The enzymes alpha-D-galactosidase, beta-D-galactosidase, beta-NAc-D-glucosaminidase alpha-L-fucosidase and neuraminidase were assayed. Considerable glycosidase activity was present in most faecal samples. Similar activities of all the enzymes assayed were found in faeces from patients with ulcerative colitis, Crohn's disease and normal controls and there was no significant correlation with disease activity. These results imply that relapse of ulcerative colitis is not initiated by increased degradation of colonic mucus by faecal glycosidases but do not exclude a role for bacterial mucus degradation in the pathogenesis of ulcerative colitis. PMID- 2991090 TI - Investigation of beta-endorphin and adrenocorticotropic hormone in placenta, amniotic membrane and umbilical cord using an immunoperoxidase technique. AB - The identification of immunoreactive beta-endorphin, beta-lipotropin and ACTH from extracts of human placentas by radioimmunoassay suggests a probable synthesis of these peptides in the placenta. In this study we investigated the presence of beta-endorphin and ACTH in placenta, amniotic membranes and umbilical cord using a peroxidase-antiperoxidase staining technique. Tissues were obtained immediately after delivery, fixed in formalin and embedded in paraffin. As control tissues autopsy specimens of pituitary glands from adults and newborns were used. All sections of pituitary glands showed a positive reaction; negative results were observed in sections of placentas, umbilical cords and amniotic membranes. We conclude that no intracellular storage of beta-endorphin and ACTH takes place in the examined tissues. PMID- 2991091 TI - Central opioid receptor activity in patients affected by puerperal, idiopathic and tumoral hyperprolactinemia. AB - The aim of our study was to evaluate the activity of opioid receptors involved in the regulation of prolactin secretion in different kinds of hyperprolactinemic states (puerperal, idiopathic and tumoral subjects). Prolactin plasma level changes were measured after the acute administration of a met-enkephalin synthetic analogue. FK 33-824, and placebo. The prolactin-releasing effect of this drug, evident in control subjects, was blunted in patients affected by idiopathic hyperprolactinemia, and absent in puerperal and tumoral patients. These results indicate that hyperprolactinemia affects the activity of central opioid receptors, with the most evident effects in patients with highest prolactin plasma levels. PMID- 2991093 TI - [Effect of a pectin-containing bran product on gastrointestinal passage and metabolism]. PMID- 2991092 TI - [Multimodal treatment of undifferentiated astrocytomas of the cerebrum. Indications for chemotherapy]. PMID- 2991094 TI - [Serum levels of pyridostigmine in myasthenia gravis: methods and clinical significance]. AB - Correlation studies on patients with myasthenia gravis are reported in which clinical assessment of fatigue and neurophysiological findings are compared to blood levels of pyridostigmine. Measurements using a high-pressure liquid chromatography method (HPLC), give reproducible results. The levels of pyridostigmine in the serum or plasma of healthy controls and of patients show no essential differences. Components of coffee, tea, chocolate and cigarettes can markedly disturb the chromatography by adding additional peaks, so that interpretation becomes difficult or impossible. Blood levels can be measured approximately one hour after oral intake of 60 mg pyridostigmine. Concentrations rise for two to four hours and then decline exponentially. The half-life of pyridostigmine was between 156 and 210 minutes. Despite identical oral dosages, the concentration differed intraindividually and interindividually among patients. While the blood level does not reach its maximum value for 1-1 1/2 to 3 hours, the maximum clinical and neurophysiological effect of pyridostigmine appears 30-60 minutes after ingestion. Variable distribution of cholinesterase inhibitors over the different compartments (blood, synaptic region) is assumed to cause this temporal lag. If the total amount of pyridostigmine is divided into 4 5 doses, the concentration profiles over the course of a day are relatively stable. There is no significant correlation between the variations in blood level throughout one day, and changes in myasthenic symptomatology. Effects of pyridostigmine can be measured at levels as low as 5 ng/ml; at levels above 40 ng/ml further improvement can be detected only rarely. Blood levels were lower if corticosteroids were administered simultaneously; azathioprine had no influence on blood levels. Blood levels assays allow better differentiation of cholinergic and myasthenic crises and the identification of disturbed absorption and interactions with other medications. PMID- 2991096 TI - Binding and receptors for human albumin polymers and IgM/HBs circulating complex in HBsAg chronic carriers. AB - Binding activity for polymerized human serum albumin (pHSA-binding), studied in passive haemoagglutination, receptors for polymerized human serum albumin (pHSA receptors), studied in ELISA, as well as the circulating IgM/HBs complex were tested in 71 chronic carriers of HBsAg with and without liver pathology. We found that 73.2% of the sera were reactive for pHSA-binding while 45% were reactive for pHSA-receptors and 42.2% for the circulating IgM/HBs complex. The distribution and mode of association of these 3 markers showed a close correlation with the e antigen in circulation (HBeAg) and with liver disease (p less than 0.05). We further observed that pHSA-binding can be present in the absence of pHSA receptors, suggesting the possible existence of further reactants in the serum pHSA reaction. We did not observe any correlations between the circulating IgM/HBs complexes, pHSA-binding and pHSA-receptors. Blocking experiments, in fact, confirmed the non-involvement of polymerized human serum albumin in the formation of the circulating IgM/HBs complex. Elution experiments showed that, in addition to HBsAg, class-G immunoglobulins are also directly involved in the serum-pHSA reaction. PMID- 2991095 TI - [Therapy of dilated cardiomyopathy with digitalis, diuretics and vasodilators]. AB - The treatment of dilated cardiomyopathy is primarily concerned with that of congestive heart failure. Digitalis is widely use in dilated cardiomyopathy but an improvement in the prognosis has not yet been demonstrated. Furthermore, the effects of digitalis in patients with sinus rhythm are debatable. If dilated cardiomyopathy induces atrial fibrillation and tachyarrhythmia, digitalis should be used. Diuretics are helpful in the treatment of congestive heart failure associated with dilated cardiomyopathy. By reducing hypervolemia and by venous dilatation, diuretics lower preload and afterload. This leads to relief of congestion and termination of the vicious cycle of congestive heart failure. Accordingly, the prognosis of dilated cardiomyopathy might be improved by diuretics. There are numerous diuretics acting differently on the renal tubules, the choice of which depends on the renal function and serum electrolyte concentrations. Reduction of preload and afterload improves congestive heart failure as has been demonstrated repeatedly. Many substances have therefore been used for arterial and venous dilation with differing results. At least for short term periods, congestion is reduced and cardiac output increases. Especially inhibitors of angiotensin II converting enzyme are very effective since they act both in the arterial and venous systems. Additionally, inhibition of the action of angiotensin may be regarded as causal therapy since the renin-angiotensin system is the trigger for vasoconstriction and fluid retention in congestive heart failure. Unlike other substances, ACE inhibitors have been demonstrated to improve prognosis of patients with congestive heart failure. At present, combined diuretic therapy and angiotensin conversion enzyme inhibition would seem the most reasonable treatment for patients with dilated cardiomyopathy and sinus rhythm. If atrial fibrillation and tachyarrhythmia develop, additional digitalis therapy is effective. PMID- 2991097 TI - [Cyclic AMP response to glucagon after partial hepatectomy]. AB - To clarify the functional recovery of the liver after partial hepatectomy, serum c-AMP responses to glucagon were investigated after hepatectomy in rats. Serum glucagon, insulin and weight of the regenerating liver were simultaneously measured. Wistar strain male rats, weighing 120-130 g, were divided into three groups; I 30% hepatectomy, II 70% hepatectomy and III sham operation. On the first postoperative day, cyclic AMP response to glucagon was remarkably decreased in I and II groups. In group I, c-AMP response returned to the normal level 2 weeks after operation. In group II the response returned to normal at the sixth postoperative week. The liver weights were almost normal after two weeks in the both groups. Levels of serum glucagon sharply increased 24 hours after operation and returned to the normal level after 7 days. However the levels of serum insulin showed no remarkable changes. In conclusion, the complete functional recovery of the liver after hepatectomy needed 6 weeks in cyclic AMP response to glucagon and was delayed 4 weeks to that of the liver weight after 70% hepatectomy. Insulin seems to have no direct effect of the liver regeneration. PMID- 2991098 TI - [Studies on adipocyte glucagon receptor assay--with special reference to the effect of cold acclimation on glucagon receptors of white adipocytes]. AB - The changes in glucagon receptors of white adipocytes from cold-acclimated rats were investigated to know the metabolic role of glucagon in cold acclimation by establishing a glucagon radioreceptor assay system for isolated white adipocytes. Glucagon radioreceptor assay methodology The binding of 125I-labelled glucagon to isolated epididymal white adipocytes was linearly related to the number of cells (0.5-2.0 X 10(5) cells/ml) added in the medium. At a cell concentration higher than 3.0 X 10(5) cells/ml, the amount of specific binding failed to show the proportional relationship to the number of adipocytes. The effects of incubation temperature (4 degrees C, 25 degrees C and 37 degrees C) on the glucagon binding were investigated. Incubation at 25 degrees C was adopted in the present study because of the highest maximum binding and the longest steady state obtained. Preincubation at 25 degrees C for 15 min increased significantly the amount of specific binding. It was confirmed that bacitracin, polypeptide antibiotics, inhibited significantly the degradation of glucagon. The glucagon binding under these conditions was found to be saturable and reversible, validating a specific reaction for the glucagon receptor. When a Scatchard plot was constructed, the data was curvilinear with an upward concavity, indicating the presence of at least two classes of binding site with different fixed affinities or of negatively cooperative interactions between receptors. It was concluded that an appropriate condition for glucagon receptor assay of white adipocytes consists of cell concentration of 1 X 10(5) cells/ml, 15 minute-preincubation and 30 minute reaction at 25 degrees C in the presence of bacitracin (1 mg/ml). Effect of cold acclimation on glucagon receptors of white adipocytes Cold acclimation decreased the size and increased the number of epididymal white adipocytes. Cold acclimation increased the number of glucagon receptors of white adipocytes; about 140% increase expressed as per cell, approximately 260% increase per unit of surface area and 210% increase per whole tissue. The affinity of binding sites was not changed. The increased binding sites could explain, at least partly, the enhanced metabolic response of cold-acclimated rats to glucagon. PMID- 2991099 TI - [A case of Krukenberg tumor associated with elevation in blood testosterone]. AB - One case of Krukenberg tumor associated with elevation in blood testosterone levels was reported. The patient was a 31-year-old woman. She visited the outpatient service of our hospital with the chief complaint of uterine bleeding and infertility. She was diagnosed to have an anovulatory cycle associated with elevation in blood testosterone levels. However, after an examination at our hospital, a tumor of the left ovary was pointed out and she underwent bilateral ovariectomy. Postoperatively, tarry stool was passed, for which she received an examination at the department of internal medicine. Roentgenography of the stomach and examination by a gastric camera revealed undifferentiated carcinoma of Borrmann's type III in the greater curvature of the angular area. This lesion was diagnosed to be Krukenberg's tumor and a gastrectomy was performed. On the other hand, the blood testosterone level which had shown elevation before resection of the ovaries was restored to the normal range after the operation. Various reports have been published concerning the endocrinological activity of this tumor. It remains to be elucidated whether it is due to metastasis of the carcinoma or if it originates from other ovarian parenchyma. However, it has been reported that elevation in endocrinological activity is apt to occur when the tumor originates from the digestive tract. PMID- 2991101 TI - Deranged metabolism of cyclic nucleotides in liver diseases. AB - To clarify the factor(s) responsible for changes in the plasma cyclic GMP concentration in liver diseases, we measured the plasma levels of cyclic GMP, along with cyclic AMP, in various clinical stages of chronic liver diseases and acute hepatitis. The level of cyclic GMP was found to increase significantly in the early stage of acute hepatitis, in the decompensated stage of liver cirrhosis, and in malignant diseases. In the former two states, it is postulated that decreased hepatic mass is responsible for the changes in the plasma cyclic GMP concentration. The retention rate of indocyanin green (ICGR15) was highly correlated with the plasma cyclic GMP level. The result suggests that the determination of plasma cyclic GMP is useful as an index of the reserve function of the liver in disease states. PMID- 2991100 TI - Effect of resection of small intestine on the interaction of vasoactive intestinal peptide with rat colonic epithelial cells. AB - Vasoactive intestinal peptide (VIP) receptors and VIP-dependent cyclic AMP production were studied in rat colonic epithelial cells 3 days after a 60% resection of the small intestine. Basal cyclic AMP levels were similar in both control and resected animals. The potency, but not the efficiency, of the peptide on the stimulation of cyclic AMP production was diminished in cells from resected rats. Accordingly, the affinity of VIP receptors, but not the binding capacity, decreased as a consequence of the loss of a part of the small intestinal mucosa. These observations are consistent with the known inhibitory role of cyclic AMP on cell proliferation in colonic epithelium and other tissues and suggest a participation of VIP acting through the cyclic nucleotide in the compensatory hyperproliferative response of the colon following massive resection of the small intestine. PMID- 2991102 TI - Depression, masochism, and biology. AB - In examining the conditions of depression and masochism, my intention has been to expand our area of study beyond psychodynamics alone. My first aim was to present what I believe are the intimately intertwined dynamics of each condition, a metaphorical double helix of depression and masochism in a matrix of narcissism. It may make clearer how either depression or masochism may present clinically in combination, at times in tandem, or manifestly as either state alone. My second, but major, aim is ecumenical: to interweave contributions from outside psychoanalysis. The neurophysiological bases and genetic determinants for most depressions are by now well-recognized. Masochism, much like depression, with which it is closely allied, may not necessarily arise out of conflict alone. I have presented brief excerpts of material, much of it still speculative, from areas of genetics, biochemistry, and ethology, to support the concept of a biological anlage for masochism. This would help explain the enormous difficulties therapists find in the path of its successful treatment. I believe Lorenz's theories on animal "bonding" suggest precursors to our concepts of masochism. I further believe our field of study has reached the point at which these and probably additional scientific disciplines can be helpful or even necessary for the further understanding of character, and for the solution of the persistent riddle of masochism, whose full understanding has continued to elude us. PMID- 2991104 TI - Time course and mechanism of alterations in proton relaxation during liver regeneration in the rat. AB - We studied the proton T1 and T2, water and lipid content of regenerating rat liver from 1 to 7 days after 70% hepatectomy. Liver from normal and sham-operated animals and splenic tissue from all animals were studied as controls. In vivo proton spectroscopy and imaging of liver was performed in a separate group of control and posthepatectomy rats. The T2 of regenerating liver, but not of sham or normal control liver, was prolonged. Changes in T1, relative to normal tissue, were found in liver and spleen of both operated groups. Lipid content, assessed both by extraction of tissue samples and by in vivo spectroscopy, was increased in regenerating tissue but not in controls. Water content was similarly increased in regenerating liver tissue. Changes in water and lipid content appeared to contribute to the alterations in proton relaxation which we observed. PMID- 2991103 TI - Effect of chronic ethanol administration on enzyme and lipid properties of liver plasma membranes in long and short sleep mice. AB - Mechanisms of alcoholic liver disease are still ill defined. We evaluated in two outbred lines of mice whether chronic ingestion of ethanol alters the lipid composition and/or enzyme activity of liver plasma membranes. Two mouse lines with different sensitivities towards the hypnotic effect of ethanol, designated long sleep and short sleep, were fed a liquid diet containing ethanol for 30 days. Ethanol intake reached 30 gm per kg per day in both lines, and serum ethanol levels were similar. In addition, hepatic triglyceride levels were similarly increased 2-fold with ethanol feeding. The following effects of ethanol treatment were observed in liver plasma membrane fractions: (i) Na+,K+-ATPase was significantly increased to 26% above control in long sleep only; (ii) alkaline phosphatase activity was 2-fold increased in both lines; (iii) 5'-nucleotidase, leucine aminopeptidase and Mg2+-ATPase activities remained unchanged in both lines; (iv) unesterified cholesterol and total phospholipid contents were unaltered in both lines, and (v) cholesteryl esters were increased in both lines, but to a greater extent in short sleep (1.5 vs. 4-fold). Thus, chronic ethanol ingestion induces specific alterations in liver plasma membrane structure and function, suggesting that adaptive responses to ethanol may be determined in part by inherited factors. PMID- 2991105 TI - The spectrum of complement-fixing antinuclear antibodies in patients with hepatocellular carcinoma. AB - Sera from 230 hepatocellular carcinoma patients were tested for antinuclear antibodies by anticomplement immunofluorescence in 16 types of transformed, diploid or primary cells of human, monkey, chimpanzee or rat origin. As controls, we tested 85 sera from patients with chronic liver diseases, 48 sera from patients with nonhepatic cancers and 164 sera of normal controls. Exactly 11.2% of all cancer patients but only 3.6% of noncancer patients had complement-fixing antinuclear antibody that reacted with all substrates. Only sera from hepatocellular carcinoma reacted with subsets of the tumor cell substrates. These sera reacted with hepatocellular carcinoma cells and nonhepatic cancer cells (antitumor) or only with one or more of the human hepatocellular carcinoma cell lines, PLC/PRF/5, Hep3B and Mahlavu, that were derived from HBsAg-positive patients (antihepatocellular carcinoma). Three of these reacted only with hepatitis B virus DNA-positive cells (PLC/PRF/5 and Hep3B) that contained "hepatitis B-associated nuclear antigen," 1 reacted only with hepatitis B virus DNA-negative Mahlavu cells, 1 reacted with PLC/PRF/5 and Mahlavu and 3 reacted with all 3 cells. The nuclear antigen in Mahlavu was expressed as a homogeneous fluorescence that spared the nucleoli, was present in a lower percentage of cells than hepatitis B-associated nuclear antigen and was more thermostable than hepatitis B-associated nuclear antigen. However, it resembled hepatitis B associated nuclear antigen in kinetics of expression and susceptibility to digestion with DNase, RNase and proteinase K. The nature of the nuclear antigens in the hepatocellular carcinoma cells is poorly understood but one possibility is that they may represent the expression of viral or tumor-related genes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991106 TI - A carrier-protein receptor is not a prerequisite for avid hepatic elimination of highly bound compounds: a study of propranolol elimination by the isolated perfused rat liver. AB - The highly efficient hepatic extraction of propranolol by the isolated perfused rat liver does not diminish when albumin binding is increased from 30 to 75%. One possible explanation of this insensitivity of propranolol uptake to changes in albumin binding is the mediation of uptake of bound ligand by an albumin receptor on the hepatocyte as postulated for oleate, taurocholic acid and rose bengal. To test this hypothesis, the hepatic extraction of propranolol was studied in the isolated perfused rat liver using alpha 1-acid glycoprotein, which lacks a hepatocyte receptor, as the carrier protein in the perfusate rather than albumin. Livers were perfused with a medium containing propranolol (4 microM) and varying concentrations of alpha 1-acid glycoprotein (0 to 25 microM). Hepatic extraction of propranolol was very high (0.990 +/- 0.006; mean +/- S.D.) and did not alter significantly despite an increase in bound fraction from 0.2 to 0.8, thus closely paralleling the findings when albumin is the carrier protein. This result indicates that bound propranolol is efficiently cleared by the liver, presumably by a "free intermediate" mechanism, in the absence of a specific carrier-protein receptor on the hepatocyte. This study does not, therefore, support the albumin receptor hypothesis. PMID- 2991107 TI - Acute and chronic changes in the microcirculation of the liver in inbred strains of mice following infection with mouse hepatitis virus type 3. AB - The acute and chronic effects of mouse hepatitis virus type 3 on the microcirculation of the liver in both semisusceptible C3HeB/FeJ and fully resistant A/J mice were studied. In the C3HeB/FeJ mice, abnormalities of microcirculatory flow were noted as early as 12 hr after infection and by 24 hr, localized avascular foci appeared. Disturbances were characterized by granular blood flow, sinusoidal microthrombi, distortion of sinusoids by edematous hepatocytes and necrotic lesions. Following the acute infection, Day 10, two patterns of chronic disease were observed. Eighty percent of the mice developed chronic granulomatous hepatitis whereas in the remaining 20% a more severe chronic aggressive hepatitis was observed which was characterized by ongoing hepatocellular necrosis and a marked mononuclear cell infiltrate. In both cases, in vivo microcirculatory abnormalities were found predominantly around visible lesions. Onset of the microcirculatory abnormalities was found to be concomitant with a rise in monocyte related procoagulant activity. Procoagulant activity rose acutely and remained elevated throughout the chronic phase but was higher in animals with severe disease. In contrast to the above, normal blood flow and histology were seen in the resistant A/J mice at all times following infection, and procoagulant activity remained at basal levels despite active viral replication as demonstrated by immunofluorescence studies and recovery of infectious virus. These observations suggest a role for monocyte procoagulant activity in the development of microcirculatory abnormalities following mouse hepatitis virus type 3 infection which may be important in the pathogenesis of the disease. PMID- 2991108 TI - Persistence of IgM hepatitis A antibody after clinical type A hepatitis. PMID- 2991109 TI - Beyond gamesmanship: strategies for coping with prospective payment. AB - Prospective payment represents a major economic challenge to inpatient psychiatric units. For a unit to survive, increased efficiency of treatment will be crucial but will probably not be enough. Most units will need to alter the percentages of high-cost and low-cost patients they treat (case mix) and set less ambitious treatment goals. When altering case mix, ethical and political concerns must be considered first, followed by careful evaluation of which patients will benefit most from high-cost treatment. The authors recommend several measures for improving efficiency of treatment, including decreasing time to make dispositions, using electroconvulsive therapy when indicated, and strengthening aftercare services. They also recommend negotiating with hospital administrators to redeploy conserved resources within the psychiatry department rather than elsewhere. PMID- 2991110 TI - Multinucleated giant cells and HTLV-III in AIDS encephalopathy. PMID- 2991111 TI - The prevalence of carcinoma in situ in normal and cancer-associated breasts. AB - Two hundred ninety-two human breasts were examined in toto by a subgross sampling technique with histologic confirmation. The samples consisted of 185 breasts from random autopsies, 63 cancer-containing breasts, and 44 breasts contralateral to cancer-containing breasts. The method permits the identification and enumeration of essentially all of the dysplastic, hyperplastic, and neoplastic lesions present in each breast. Emphasis was on the prevalence within each sample category of ductal carcinoma in situ (DCIS), lobular carcinoma in situ (LCIS), and epithelial proliferative lesions with severe atypia, previously termed ALA 4 and ALB 4, which correspond to the clinicopathologic entities atypical ductal hyperplasia and atypical lobular hyperplasia, respectively. Additional primary foci of DCIS (unrelated to invasive breast carcinoma, if present) were found in 52.5 per cent of cancer-containing breasts, and were seen in 47.7 per cent of contralateral and 5.9 per cent of the breasts from random autopsies. Lobular carcinoma in situ was generally seen only in association with infiltrating carcinoma, usually of the ductal type. No LCIS was seen in the breasts from random autopsies. These trends are the same if the proliferative lesions with severe atypia are included with carcinoma in situ. The numbers of lesions were also markedly greater in affected cancer-associated breasts than in affected breasts obtained from autopsies. These findings suggest that LCIS, although a rare lesion in the general population, may be a significant marker for clinical carcinoma. They support previous studies showing a small percentage of women with undetected DCIS of uncertain clinical and biological potential. The multicentric nature of preinvasive breast carcinoma is further substantiated. Finally, when the prevalence and number of lesions are considered in association with the ages of the patients, the lower prevalence of such lesions in the older patients in each sample suggests that at least some DCIS and LCIS may be dependent on a premenopausal hormonal milieu for their continuing existence. PMID- 2991112 TI - Malignant bronchial oncocytoma: case report and review of the literature. AB - An oncocytoma of the bronchus with an infiltrative growth pattern and a lymph node metastasis were examined by light and electron microscopy. Lobectomy, with removal of all tumor-bearing tissue, was the only treatment. Two years after surgery the patient had no local recurrences or metastases. PMID- 2991114 TI - Human lens gamma-crystallin sequences are located in the p12-qter region of chromosome 2. AB - The human gamma-crystallin genes constitute a multigene family whose members are only expressed in the eye lens. The chromosomal location of these sequences has been determined by screening a panel of human/rodent hybrid cell lines containing overlapping subsets of human chromosomes for the presence of human gamma crystallin sequences. By correlating these genomic hybridization data with the chromosomal constitution of the somatic cell hybrids, all human gamma-crystallin sequences could be assigned to chromosome 2. The use of human/hamster cell hybrids derived from human Burkitt lymphoma cells carrying a reciprocal translocation between human chromosomes 2 and 8, allowed a further localization of the sequences to the region 2p12-qter. PMID- 2991113 TI - Genetic analysis of carbamyl phosphate synthetase I deficiency. AB - Carbamyl phosphate synthetase I deficiency (CPSD) is an autosomal recessive disorder of ureagenesis characterized by hyperammonemic coma in the neonatal period. To study the genetic basis of CPSD we have performed a molecular analysis of the CPS I genes in CPSD patients from six unrelated families. Using a cDNA probe for the human CPS I gene and restriction endonuclease mapping techniques, we observed no abnormality in the number of size of the hybridizing DNA fragments from the seven affected individuals examined. These findings suggest that no gross alteration affected the CPS I genes. We did detect a frequent restriction fragment length polymorphism (RFLP) at the CPS I locus which we employed as a linkage marker. Our results suggest the polymorphic CPS I restriction fragments cosegregate with the CPSD phenotype, and that linkage disequilibrium exists between the CPSI RFLPs studied and the affected alleles. The RFLPs described may enable prenatal detection of CPSD in families where the coupling phases between CPSD alleles and RFLPs can be determined. PMID- 2991115 TI - Linkage studies of X-linked mental retardation: high frequency of recombination in the telomeric region of the human X chromosome (fragile site/linkage/recombination/X chromosome). AB - One of the commonest forms of X-linked mental retardation is associated with a fragile site at Xq27 on the human X chromosome which can be visualised structurally after culturing cells in folate-deficient media. Unusually, the mutation can be transmitted through a phenotypically normal male. There is already some evidence that the gene loci for G6PD and factor IX are linked to this mental retardation locus. We have followed the inheritance of a DNA sequence 52A, in fragile site families that are also informative for factor IX. We demonstrate that these probes are localised at Xq27/Xq28-Xqter, close physically to the fragile site. We did not find close linkage between 52A, factor IX, and the fragile site in the families studied despite 52A and factor IX showing linkage in normal families. We discuss the importance of these data for the genetic mapping of this region of the human X chromosome and the implication for the use of these DNA probes for clinical diagnosis. PMID- 2991116 TI - The second human calcitonin/CGRP gene is located on chromosome 11. AB - A second human calcitonin/calcitonin gene related peptide (hCT/CGRP) gene has been identified. This second hCT/CGRP gene has been shown to contain sequences highly homologous to exons 3, 5 (CGRP-encoding), and 6 of the first hCT/CGRP gene, but sequences closely related to exon 4 (CT-encoding) could not be demonstrated. Southern blot hybridization analysis of DNA from human-rodent somatic cell hybrids showed that the second hCT/CGRP gene is located in the q12 pter region of chromosome 11. The first hCT/CGRP gene has previously been assigned to the p13-p15 region of chromosome 11. PMID- 2991118 TI - High-resolution studies in patients with aniridia-Wilms tumor association. PMID- 2991117 TI - The apolipoprotein CII gene: subchromosomal localisation and linkage to the myotonic dystrophy locus. AB - The human apolipoprotein CII gene probe detects a restriction fragment length polymorphism located on chromosome 19. We have investigated the linkage of this polymorphism to the myotonic dystrophy locus in families. The two loci are closely linked with a maximum Lod score of 7.877 at 4% recombination. The close linkage and informativeness of the APOC2 polymorphism suggest that this probe may be of use for presymptomatic diagnosis of the myotonic dystrophy gene. The APOC2 gene was localised to the region 19p13-19q13 using somatic cell hybrids, providing further evidence that the myotonic dystrophy locus is situated in the central region of chromosome 19. PMID- 2991119 TI - A mitotic recombination in Wilms tumor occurs between the parathyroid hormone locus and 11p13. AB - Wilms tumor is believed to occur as the result of two mutations affecting both alleles of a critical gene located within the p13 band of chromosome 11 (Knudson and Strong 1972; Riccardi et al. 1978). Several mechanisms by which these mutations occur have already been determined in retinoblastoma (Cavenee et al. 1983) and Wilms tumor (Koufos et al. 1984; Orkin et al. 1984; Reeve et al. 1984; Fearon et al. 1984a; Eccles et al. 1984). Of the various mechanisms, however, no example of a mitotic recombination was demonstrated in Wilms tumor. An example is presented here which has been detected by the use of restriction fragment length polymorphisms (RFLPs) mapping to chromosome 11p. In addition the data presented are consistent with the mapping location of parathyroid hormone (PTH) being proximal to 11p13. PMID- 2991120 TI - The microbial aetiology of genital ulcers in black men in Durban, South Africa. AB - The microbial aetiology of genital ulcers was assessed in 100 black men attending a sexually transmitted disease (STD) clinic in Durban, South Africa. Forty patients harboured Haemophilus ducreyi, one hepes simplex virus, and one Neisseria gonorrhoeae. Syphilis was diagnosed in 44 patients on the basis of dark field microscopy or positive syphilis serology test results, or both. Of these 44 patients, eight also harboured N ducreyi, one herpes simplex virus. Lymphogranuloma venereum was diagnosed in one patient. No cause of ulceration could be found in the remaining 16 patients. PMID- 2991121 TI - Psychological support and counselling for patients with acquired immune deficiency syndrome (AIDS). AB - The growing numbers of patients with the acquired immune deficiency syndrome (AIDS) in England had led to the development of a counselling service for patients, their sexual partners, contacts, and others. The nature of the counselling is described with reference to the primary qualitative issues arising from diagnosis. The necessity for preventative education for patients and those at risk is stressed, and guidelines for low risk sexual activities are outlined. PMID- 2991123 TI - Polynuclear aromatic compounds, Part 4, Bitumens, coal-tars and derived products, shale-oils and soots. PMID- 2991122 TI - Sodium polyanethol sulphonate discs to identify Gardnerella vaginalis. PMID- 2991124 TI - Shale-oils. PMID- 2991125 TI - Regulation of immunoglobulin secretion by factor H of human complement. AB - As human B lymphocytes and macrophages carry surface receptors for Factor H (B1H), we investigated the possibility that this complement component regulates their function. Factor H inhibits immunoglobulin secretion by peripheral mononuclear cells (MNC) stimulated with pokeweed mitogen if present at the initiation of the cultures and at concentrations greater than 50 micrograms/ml. Factor H also inhibited stimulation and differentiation of purified B cells into immunoglobulin-secreting cells by Epstein-Barr virus (EBV). The inhibitory effect of Factor H was abrogated if anti-Factor H antibody was present in the cultures. EBV-transformed B-cell lines secreted less immunoglobulin if Factor H was present in the culture for at least 4 days. Culture of MNC with Factor H did not lead to the generation of suppressor T cells or macrophages. In contrast, Factor H did not cause proliferation of human peripheral total MNC or enriched T-cell or B cell subpopulations. Also, Factor H did not inhibit the proliferation of MNC in response to several mitogens and antigens. Our results strongly indicate that Factor H is able to block human B-cell differentiation in vitro without blocking the proliferative ability of the cells. Factor H seems to act directly on the B cells through its receptor on their surface, since it inhibited T-dependent and T independent B-cell differentiation but generated no suppressor cells. PMID- 2991126 TI - Influences of nutrition on immunity and susceptibility to mouse hepatitis virus type 2. AB - Resistance to mouse hepatitis virus (MHV) in C3H mice is a genetic trait which appears 3-4 weeks after birth. However, when these animals were weaned on a low protein diet (8% casein), they remained susceptible to MHV-2 infection until they reached 8-9 weeks of age. During this period, the protein-restricted C3H mice were as susceptible to MHV-2 as the genetically susceptible congenic C3Hss strain. The delay in the emergence of resistance in the protein-restricted mice could be corrected by injecting these animals with spleen cells from 6-week-old C3H mice. Thymocytes from normal C3H mice, and splenocytes and thymocytes from protein-restricted C3H mice, were not protective. However, spleen cells from the protein-restricted mice were more responsive to phytohaemagglutinin, lipopolysaccharide and concanavalin A than spleen cells from normal C3H. The enhanced lymphoproliferative response in spleen cells from protein-restricted mice was abrogated by the addition of plastic-adherent cells obtained from normal C3H spleens. Spleen cells from protein-restricted and from genetically susceptible C3Hss mice also possessed more spontaneous cytotoxicity against MHV infected 3T3 fibroblasts. PMID- 2991127 TI - Fc receptor-mediated desensitization of superoxide (O2-) generation response of guinea-pig macrophages and polymorphonuclear leucocytes. AB - Guinea-pig macrophages were pretreated with soluble immune complexes (1 hr, at 37 degrees). By this procedure, the capacity of the cells to produce superoxide anions (O2-) upon stimulation with wheatgerm agglutinin (WGA) was inhibited by as much as 80% (desensitization). However, the receptors for WGA were still available on the cell surfaces as determined by the binding of 125I-labelled WGA to both the pretreated and control cells. This inhibition of O2- generation was also observed when opsonized zymosan (Op-zymosan) was used as the stimulus for O2 generation. In contrast, the inhibition by soluble immune complexes was less than 25% of the control levels when the desensitized cells were stimulated with N formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) or phorbol myristate acetate (PMA). The same procedure was also used to desensitize guinea-pig polymorphonuclear leucocytes (PMN). In this case, the O2- generation of the PMN was suppressed to 50-55% of the control level when WGA or fMLP was used as stimulus. However, the PMA-induced O2- generation was far less affected by this treatment, as was the case with the macrophages. The activity of the desensitized macrophages for O2- generation recovered gradually to a normal level 24 hr after removal of the immune complexes from the medium. These results suggest that (i) some regulatory mechanisms which suppress the activation of NADPH oxidase are accentuated by prolonged exposure of guinea-pig macrophages and PMN with soluble immune complexes; (ii) the mechanism for O2- generation mediated by WGA or Op zymosan may be different from that for O2- generation mediated by PMA, and (iii) the mechanism for O2- generation mediated by fMLP differs between macrophages and PMN. PMID- 2991128 TI - Erythrocyte lysis by PMA-triggered neutrophil polymorphonuclears: evidence for an hypochlorous acid-dependent process. AB - Human red blood cells (HRBC) were efficiently lysed when incubated with neutrophil polymorphonuclears (PMN) in the presence of phorbol-myristate-acetate (PMA), as detected by a 4-hr 51Cr release assay. The lysis was virtually absent in the presence of catalase, azide or cyanide and in the absence of chloride ions. These findings indicate the involvement of the myeloperoxidase (MPO) hydrogen peroxide (H2O2)-chloride (Cl-) system in the cytolytic process. As the MPO-H2O2-Cl- system is capable of generating the powerful oxidant hypochlorous acid (HOCl), cytotoxicity assays were performed in the presence of taurine, glycine, serine and valine to scavenge this potentially lytic agent. Each of these compounds efficiently inhibited the HRBC lysis by PMA-triggered PMN, as well as the lysis caused by HOCl in a cell-free system. Thus, the results suggest that HOCl, or an agent with similar reactivity, plays a key role in the PMA dependent PMN-mediated cytotoxicity against HRBC targets. PMID- 2991129 TI - Induced type-B reticulum cell neoplasia of CBA mice. II. Functional similarities between tumorigenic reticulum cells and normal accessory cells. AB - In a previous report, it was proposed on the basis of phenotypic evidence that the population of transplantable cells which constituted the major component of experimentally induced type-B reticulum cell neoplasms of CBA strain mice could be a neoplastic counterpart of lymphoid dendritic or veiled accessory cells. This has been further tested by an in vitro examination of their accessory function. Both similarities and differences in behaviour between the neoplastic cells and accessory cells prepared from spleens of normal mice were found. The possible significance of these properties is discussed in relation to the process of neoplastic change. PMID- 2991130 TI - Some early events in the primary mitogenic stimulation of lymphocytes differ from later interleukin stimulation and other quiescence to growth activation systems. AB - The requirement for activity of the enzyme ADP-ribosyl transferase (ADPRT) and changes in single-strand DNA breaks were assessed during the initial stimulation of quiescent murine splenic lymphocytes with mitogen alone, the stimulation of activated blasts with IL-2-containing medium and, for comparison, the serum stimulation of quiescent fibroblasts and the induction of haemoglobin synthesis in an erythromyeloid cell line K562. Inhibitors of ADPRT, at concentrations previously found to have no effect on the proliferation of lymphoblastoid cell lines, blocked the stimulation of spleen cells by Con A or LPS; non-inhibitory analogues had much less effect. No early increase in ADPRT activity after mitogenic stimulation was detectable. The rejoining of single-strand breaks was observed after stimulation of splenic lymphocytes with Con A, but not consistently with LPS. Conversely, ADPRT inhibitors had only little effect on the IL-2-induced stimulation of Con A blasts, and no effect on the stimulation of fibroblasts or K562. Neither were any changes in strand breaks associated with these systems. These findings implicate ADPRT activity and the rejoining of strand breaks in the early mitogenic response as being distinct from later IL-2 activation and changes from quiescence to growth in other cell types. PMID- 2991131 TI - Major histocompatibility complex of the mole-rat. II. Restriction fragment polymorphism. AB - The major histocompatibility complex (Mhc) is a group of loci coding for lymphocyte membrane glycoproteins that provide the context for the recognition of foreign antigens in the initial phase of the immune response. The complex contains a large number of loci, some of which are highly polymorphic. The complexity and polymorphism pose a number of questions concerning the evolution of the Mhc. In an attempt to answer some of these questions, we have begun to study the Mhc of the mole-rat, Spalax ehrenbergi, a rodent representing a complex of sibling species occupying ecologically and geographically clearly delineated regions within the borders of Israel. In an earlier publication we identified the Spalax major histocompatibility (Smh) complex serologically and biochemically. Here, we analyze the Smh by Southern blotting of DNA fragments produced by restriction enzyme digestion. The fragments were hybridized to mouse probes specific for class I, class II, and C4 genes. The analysis has revealed that the Smh complex contains as many class I genes as the mouse does and that these genes are polymorphic. The number of class II genes could not be determined with certainty, but it is probably not greater than in the mouse. Polymorphism was also detected at the loci coding for the complement component 4 (C4), which are probably closely linked to the Smh complex. The polymorphism of mole-rat class I loci contrasts with the reported monomorphism of these loci in the Syrian hamster. Since the mole-rat leads a solitary, subterranean life, as the Syrian hamster does, ecology cannot be an explanation for the lack of class I polymorphism in the latter species. PMID- 2991133 TI - Association of class II HLA-DQ beta chain DNA restriction fragments with multiple sclerosis. PMID- 2991132 TI - Class I and class II restriction pattern polymorphisms associated with independently derived RT1 haplotypes in inbred rats. AB - This communication reports the DNA level identification of class I and class II sequences associated with 20 RT1 haplotypes which have been assigned previously to eight RT1 groups. Sixteen to 22 bands in genomic blots hybridized with the mouse pH-2III class I cDNA probe. Only the three RT1k haplotypes associated with identical class I restriction fragment patterns. Differences in restriction bands between putatively identical RT1 haplotypes were either less than or equal to 6%, or greater than 50%, suggesting a relatively high level of recombination between serologically identified RT1.A genes and the majority of class I sequences. Restriction fragment patterns associated with three RT1u haplotypes differed by less than 6%. However, intra-RT1a, intra-RT1b, and intra-RT1l restriction fragment differences were between 50 and 64%. In specific cases, different RT1 haplotypes associated with identical class I restriction patterns, e.g., RT1m (MNR) and RT1d (MR); higher resolution confirmed the difference (two bands) between RT1m and RT1d. Results of hybridization with the human DC1 beta probe confirmed that the AVN RT1a and NSD RT1b haplotypes were generated by recombinations within the vicinity of the RT1.B:RT1.D regions. These results demonstrate that a previous classification of RT1 haplotypes was incomplete and did not include the majority of class I and class II sequences which distinguish RT1 haplotypes. PMID- 2991134 TI - The rat immunoglobulin kappa light chain locus is on chromosome 4. PMID- 2991135 TI - NBT dye reduction test & myeloperoxidase activity of neutrophils in pregnant & non-pregnant women. PMID- 2991136 TI - Ethanethiol sensitive & resistant antibodies to enterovirus 70 in post conjunctivitis neurological syndromes. PMID- 2991137 TI - Viral etiology of acute respiratory infections in south Indian children. PMID- 2991138 TI - Genetics of spontaneous, high-frequency loss of b capsule expression in Haemophilus influenzae. AB - We determined that the frequency of spontaneous capsule loss in Haemophilus influenzae type b is 0.1 to 0.3%. All of 10 independent capsule-deficient variants (derived from four different type b strains) were found to be missing an identical 9-kilobase EcoRI restriction fragment when probed with a cloned piece of DNA containing sequences known to be necessary for type b capsule expression. These results suggest the existence of a specific mechanism for shutting off type b capsule synthesis at a high frequency. Intranasal infection of infant rats showed that capsule loss occurred in vivo at frequencies comparable to those observed in vitro. PMID- 2991139 TI - Oxidative metabolic response and microbicidal activity of human milk macrophages: effect of lipopolysaccharide and muramyl dipeptide. AB - Mouse macrophages can be primed by exposure in vitro to the bacterial products lipopolysaccharide and muramyl dipeptide (MDP) or in vivo by injection of MDP, so that they produce more of the bactericidal agent superoxide anion (O2-) when stimulated by phagocytosis or by contact with phorbol myristate acetate (PMA). Because little is known about the physiology of human tissue macrophages, we examined release of O2- by milk macrophages obtained from 45 normal women for the ability to undergo priming for greater O2- release. In samples from the same individuals, PMA-stimulated O2- release was similar from colostrum (0 to 3 days postpartum) or from transitional milk (5 to 8 days). Release of O2- by milk macrophages was almost identical to that by blood monocytes from the same women. Milk macrophages phagocytized and killed Candida albicans relatively effectively. Incubation with lipopolysaccharide activated the macrophages in that they were primed for greater PMA-stimulated O2- release. Incubation with the adjuvant MDP or its analog 6-O-(2-tetradecylhexadecanoyl)-MDP did not prime, but incubation with a second analog, 6-O-(stearoyl)-MDP, primed the macrophage for greater O2- release. These results indicated that human tissue macrophages can be primed for greater oxidative response by exposure to bacterial products. Potential exists for the therapeutic use of such immunomodulating agents in the enhancement of host defense. PMID- 2991140 TI - Suture materials and suturing techniques. PMID- 2991141 TI - AIDS--status 1985. Implications for sperm banks. PMID- 2991142 TI - Radiosynthesis of an opiate receptor binding radiotracer: [11C]carfentanil. AB - Carfentanil, a potent opiate receptor agonist, was labeled with 11C for in vivo studies using positron emission tomography. This synthesis was completed in approximately 30 min using iodomethane as the 11C precursor. The synthesis, purification, characterization, and determination of specific activity are presented and discussed. PMID- 2991143 TI - A convenient method for the preparation of 99mTc(V)dimercaptosuccinic acid (99mTc(V)-DMSA). AB - Tc(IV)-DMSA for kidney scintigraphy has been prepared in acidic solution. Once the labeling is done in basic solution, upon addition of a small amount of NaHCO3, a mixture of 3-4 other DMSA-complexes is formed, presumably containing Tc(V). Kidney uptake in male adult rats of the 99mTc(V)-DMSA is 1.6% injected dose/g (4.9%/total organ) compared to 16.4% injected dose/g (resp. 50.3%/total organ) for 99mTc(IV)-DMSA. PMID- 2991144 TI - Lysis of type-I collagen by squamous carcinomas of the head and neck. AB - Lysis of type-I collagen by squamous carcinomas of the head and neck has been studied in freshly excised tissues, xenografts and established cell lines. Investigations with 35 freshly excised tumours showed only low levels of active and total collagenase in both carcinomas and controls. A difference became apparent when the tissues were set up in explant organ culture where a significant (p less than 0.05) increase in total collagenase was found in 13/19 tumours compared with paired control tissues over a 4-week culture period. Two xenografts showed little capacity to lyse collagen in vitro and there was only limited evidence of an increase in total collagenase after explantation and growth in organ culture. Twenty tumour cell lines showed low levels of active collagenase. Total collagenase levels were significantly increased (p less than 0.05) in 4 of the cell lines derived from cancers of the tongue; this activity was sustained in subsequent passages. Six control fibroblastoid cell lines also showed low levels of active collagenase. Levels of total collagenase were consistently high, but this activity was transient and declined in subsequent passages. Co-cultivation experiments with II tumour-cell lines and 5 fibroblastoid cell lines showed some enhanced, synergistic destruction of collagen. Parallel experiments with supernatant media from the carcinoma and fibroblastoid lines showed no enhancement, indicating that intact carcinoma cells and fibroblastoid cells are required for synergistic collagenolysis to take place. PMID- 2991145 TI - Descriptive epidemiology of colorectal cancer. AB - The purpose of this report is to present the descriptive epidemiology of colorectal cancer using the most recent data available to highlight two characteristics of the disease. First is the great variation which takes place in the frequency of this disease over geographic areas of all sizes. Colorectal cancer is common in most countries of North America and Europe, is rare in Asia and is particularly uncommon in Africa. Internationally, the variation in colon cancer is 60-fold, and within Europe there is a 4-fold difference in the incidence of colon cancer between areas with the highest and lowest rates. For cancer of the rectum, variation internationally is 18-fold and within Europe it is 3-fold. Within the United Kingdom, colon cancer is uniformly higher in the 5 Scottish Cancer Registry Regions than in their counterparts in England and Wales. Even within Scotland there is a 4-fold range of colon cancer incidence in rates, with the North and South clearly demarcated by a striking difference in colon cancer incidence in both sexes. Secondly, examination of international mortality rates for colorectal cancer demonstrates remarkable differences in trends over time between countries. In countries where colorectal cancer mortality rates were initially low, rates have increased substantially. In many countries where rates circa 1950 were moderately high, they have increased slightly or become stabilized. However, in countries such as Scotland, Canada, England and Wales and the United States, where rates were initially high, there have been gradual falls in mortality over time. PMID- 2991146 TI - Prostaglandin and hydroxyeicosatetraenoic acid synthesis by human mesenchymal tumors. AB - The metabolism of arachidonic acid was investigated by radioimmunoassay and chromatographic techniques in 5 sarcomas and one embryonal carcinoma of human origin maintained as transplantable tumors in nude mice. The results obtained indicate that: the absolute quantities of arachidonic acid metabolites produced by a given tumor varied between experiments but the overall distribution pattern of these products, in general, remained constant from passage to passage; each tumor showed a different arachidonic acid metabolite profile in quality and quantity; 2 sarcomas of the same histological type could be clearly distinguished by their arachidonic acid metabolites; the predominant product in all tumors was 12-HETE or 15-HETE, whereas thromboxane A2 was synthesized in low quantities by all tumors; PGF2 alpha was synthesized at the highest rate by an alveolar rhabdomyosarcoma; PGE2 synthesis was highest in a malignant fibrous histiocytoma; and total prostaglandin synthesis was low in the chondrosarcoma and synovial-cell sarcomas. All results reported in this study are for the complete tumor which includes both neoplastic and stromal cells. The role that these products play in the biological behavior of mesenchymal tumor cells and normal tissues of the host remains to be determined. PMID- 2991147 TI - Host cell range of adult T-cell leukemia virus. I. Viral infectivity and binding to various cells as detected by flow cytometry. AB - Adult T-cell leukemia virus is the member of a human type-C retrovirus family (HTLV) found to be associated with adult T-cell leukemia (ATL) in Japan. In our study, HTLV was isolated from the MT-2 cell line, purified on sucrose gradient and labelled with fluorescein-isothiocyanate (FITC-HTLV). The protein pattern of the virus was determined by SDS-gel electrophoresis and assured by Western blotting using ATL patient serum. Fresh human lymphocytes, separated B and T cells, mouse and rabbit lymphocytes, mouse fibroblasts, and 13 different tumor cell lines were tested in parallel for binding of FITC-HTLV and infectability by the virus. Virus binding to cell receptors was assayed by flow cytometry. Successful infection was monitored by following the expression of HTLV-determined antigen (HTLA). Most of the cells bound FITC-HTLV at levels ranging from 5% to 130% of the MT-2 cell binding. Only fresh human T, mouse and rabbit lymphocytes were infectable by cell-free virus preparations. The results demonstrate that HTLV receptors are present on different types of cells of both human and animal origin, and that infection by the virus is restricted to fewer host cells but not limited to a specific class of human lymphocytes. PMID- 2991148 TI - Production of PDGF-like growth factor by breast cancer cell lines. AB - The breast cancer cell line T47D produces factors which show mitogenic activity for 3T3 cells. Here we describe the partial purification of one of these factors and show that it has properties similar to platelet-derived growth factor (PDGF). The T47D factor is a heat-stable hydrophobic protein with a molecular weight of around 30,000, which inhibits the binding of 125I-EGF in a temperature-dependent manner. This 30 kd protein does not act synergistically with PDGF or fibroblast derived growth factor (FDGF; also PDGF-like) in stimulating DNA synthesis; moreover, like these two factors, its mitogenic activity can be inhibited by an antiserum raised against human PDGF. A PDGF-like growth factor was also found in the serum-free medium of the breast cancer cell line MDA-MB-157. Since PDGF acts on mesenchymal cells, the production of PDGF-like growth factors by breast cancer cells suggests a basis for the intense stromal reaction seen in human breast cancers. PMID- 2991149 TI - Treatment effectiveness in an adolescent chemical dependency treatment program: a quasi-experimental design. AB - This study utilizes a waiting-list control group experimental design to empirically assess the effectiveness of a residential adolescent chemical dependency treatment program. Mailed questionnaires and telephone follow-up procedures were used to collect outcome data. Treatment can be classified as an "AA-Family" model, combining behavior modification and psychoanalytic techniques. Complete abstinence from alcohol and/or drugs was used as a criterion for success. The abstinence rate for the treatment group sample (65.5%) was significantly higher than the abstinence rate for the waiting-list control group (14.3%) at the .001 level of probability. PMID- 2991150 TI - The parenteral kinetics of ampicillin/sulbactam in man. AB - The kinetics of co-administered ampicillin plus sulbactam were examined at doses being used in clinical trials of this combination of a beta-lactam antibiotic (ampicillin) and a beta-lactamase inhibitor (sulbactam). Following 15-min infusion of 2 g ampicillin plus 1 g sulbactam, peak serum concentrations of 120 micrograms/ml ampicillin and 60 micrograms/ml sulbactam were observed. The half life was approximately one hour for both drugs. Approximately 75% of the dose of sulbactam and ampicillin was recovered in urine. The data fit a two-compartment pharmacokinetic model. Intramuscular administration of 1 g ampicillin plus 0.5 g sulbactam produced mean peak concentrations of 18 micrograms/ml ampicillin plus 13 micrograms/ml sulbactam. Estimates of variability of the data in normal male subjects are provided to serve as references for clinical trials. PMID- 2991151 TI - Rhabdomyosarcomatous uterine adenosarcoma. AB - A case of rhabdomyosarcomatous adenosarcoma of the uterine cervix is described and the literature reviewed. This variant of adenosarcoma does not appear to be more malignant than other adenosarcomas. It can and should be differentiated from embryonal rhabdomyosarcoma and malignant mixed mullerian tumor because of the different prognostic implications and therapeutic approaches. PMID- 2991152 TI - A mixed germ cell-sex cord-stromal tumor of the ovary with retiform tubular structure: a case report. AB - An unusual mixed tumor of the ovary in a 7-month-old girl with a normal 46,XX karyotype, is presented. The tumor was composed mainly of small epithelial cells of sex cord type, but also contained multiple foci in which the presence of large cells resembling germ cells warranted the diagnosis of mixed germ cell-sex cord stromal tumor. An additional histologic feature was the presence in the fibrous stroma of the tumor of tubules of probable sex cord derivation arranged in a retiform pattern. These histologic features suggested that the tumor was a variant of the mixed germ cell-sex cord-stromal tumor of the ovary. PMID- 2991153 TI - Beta-endorphin does not protect alkylation of opiate receptor by N ethylmaleimide. AB - Rat brain membranes were incubated in N-ethylmaleimide (NEM, 0.5-1.0 mM) in the presence and absence of various concentrations of morphine, Leuenkephalin and human beta-endorphin (beta h-EP). After sufficient washing, the binding of dihydromorphine (DHM), [D-Ala-D-Leu]-enkephalin (DADLE) and tritiated beta h-EP was 10-40% above that of membranes treated with NEM alone. There was no additive effect of morphine and Leu-enkephalin with respect to their effect on recovery of beta h-EP binding. Evaluation of beta h-EP as protecting ligand proved to be difficult since preincubation completely inhibits subsequent DHM and DADLE binding unless a more extensive washing protocol is employed. A protocol for washing beta h-EP preincubated membranes using a Tris-phosphate buffer of pH 6 containing 150 mM NaCl, 20 mM MgCl2 and 10% glycerol was used to recover enough binding potential to evaluate the effects of beta h-EP preincubation towards NEM treatment. Preincubation with beta h-EP itself at 0.1-1.0 microM did not result in any increased recovery of opiate binding, in contrast to the findings with the other two ligands. PMID- 2991154 TI - Protection and deprotection of horse cytochrome c. AB - The last step in the semisynthesis of horse cytochrome c analogues (formation of the bond 65-66) requires the conformation of the complex between two complementary fragments, (1-65) lactone and (66-104). The fragments can be obtained from a limited degradation with cyanogen bromide. The amino component in this reaction can also be obtained from organo chemical synthesis in which the C terminal fragment (81-104) is required in a selectively protected form. The latter is available from a cyanogen bromide degradation of ubiquitously protected cytochrome c. The details of the protection/deprotection reaction and the properties of nonadecamethylsulfonylethyloxycarbonyl cytochrome c are described. PMID- 2991155 TI - Analysis of HSV isolated from patients with unilateral and bilateral herpetic keratitis. AB - Restriction enzyme analysis was utilized to investigate 11 HSV isolates from patients with herpetic keratitis. The electrophoretic profiles of the DNAs indicate that the isolates belong to HSV-1 and are epidemiologically unrelated. Two of the isolates, derived from a patient with simultaneous bilateral herpetic keratitis were further characterized by genetic and phenotypic analyses. These studies showed the following: (i) the two recrudescent lesions were caused by the same HSV-1 isolate; (ii) the isolate yielded two different plaque phenotypes; (iii) these plaques showed different sensitivity to I.D.U. PMID- 2991156 TI - Neurochemical aspects of depression: the past and the future? AB - The role of aberrant neurochemical substrates in the etiology of depression and the neurochemical mechanisms of antidepressant therapies have been the subjects of many hypotheses in the last 30 years. Pharmacological studies of early antidepressant drugs indicated that brain monoamines were significantly affected by these drugs and these led to the formulation of the biogenic amine hypothesis of depression. Although this hypothesis has been of heuristic value in the study of drug mechanisms and has provided a basis for screening drugs for antidepressant potential, deficiencies in it have become apparent. Neuroanatomical and neurochemical considerations favour the view that brain noradrenaline and serotonin systems may serve as bias adjusting systems for each other and numerous other neural systems. As a consequence of such a relationship, a primary defect in some other neural system would appear amplified in measurements of serotonin or noradrenaline. A possible site for this primary defect may be in membrane composition and function. Recent studies have found that typical and other antidepressant therapies have a pronounced effect on membrane lipids. Thus, in view of the important functions of membrane lipids and the fact that they have been linked to the initiation and development of a number of other disease processes, it is now suggested that consideration be given to them as playing primary causal roles in the etiology of depression and as a site of action for antidepressant drugs. PMID- 2991157 TI - Height increase in children with Down syndrome following treatment with psycho pharmacotherapeutic agents. PMID- 2991159 TI - Effect of tremorigenic agents on the cerebellum: a review of biochemical and electrophysiological data. PMID- 2991158 TI - Neural plasticity and recovery of function after brain injury. PMID- 2991160 TI - Multiple synaptic receptors for neuroactive amino acid transmitters--new vistas. PMID- 2991161 TI - Liver cell adenoma presenting as acute abdomen. AB - A 31-year-old female presented with a sudden onset of acute abdominal pain in the right hypochondrium. Two days later, the patient was in shock and suffering from severe intra-abdominal bleeding. Investigations showed that the bleeding originated in the right lobe of the liver. The patient had been taking oral contraceptives for seven years. She underwent a laparotomy and right lobectomy of the liver which was performed successfully for bleeding cell adenoma. The patient made a full recovery. PMID- 2991162 TI - Lead and mercury toxicity and the rod light response. AB - Lead and mercury have been reported to alter selectively the rod component of the electroretinogram, and to inhibit the phosphodiesterase in rod outer segments which may be responsible for generating the rods' light response. The authors have investigated the effect of lead and mercury on the voltage response to light of rods, and compared these effects with those of the phosphodiesterase inhibitor papaverine. Lead and mercury, like papaverine, slow the light response. In addition, papaverine increases the light response amplitude while lead decreases it. Mercury initially increases and then decreases the amplitude. The late decrease in amplitude produced by mercury is associated with rod degeneration: an effect which may mimic degenerative diseases in which the rod phosphodiesterase is insufficiently active. These results demonstrate that the changes of electroretinogram induced by lead and mercury can be accounted for by the changes in receptor potential these heavy metals produce. The changes in receptor potential seen are consistent with mercury inhibiting the rod phosphodiesterase, and with lead having an action in addition to phosphodiesterase inhibition. PMID- 2991164 TI - Prolactin-induced polyamine biosynthesis in spleen and thymus: specific inhibition by cyclosporine. AB - The induction of ornithine decarboxylase (ODC) in the rat spleen and thymus in response to prolactin shows a dose-dependent sensitivity to cyclosporine (CsA), a known immunosuppressive drug. Marked inhibition of prolactin-stimulated ODC activity occurred at 0.12 mg CsA/kg body weight, and nearly total inhibition was detected at 1.2 mg CsA/kg, a dose comparable to that used to suppress organ rejection processes. CsA blocked ODC induction in response to prolactin injection in both intact and hypophysectomized rats, suggestive of a direct effect of prolactin on spleen and thymus. Furthermore, we were able to demonstrate specific 125I-prolactin binding sites on the rat spleen and thymus. Although growth hormone was capable of elevating ODC activity in spleen, this elevation was not inhibited by CsA. ODC activity in response to cyclic AMP-mediated hormones and to steroid analogs such as dexamethasone was not affected by CsA. Agents known to alter calcium channeling mostly had random enhancing capacity when administered with CsA. The most potent elevation occurred in spleens in response to aminophylline plus CsA. It should be noted, however, that aminophylline alone slightly inhibited the basal level of ODC in the spleen. Insulin, which elevated ODC in the thymus and spleen in a dose-dependent manner, also was not affected by concurrent CsA administration. Prolactin administration altered the RNA and DNA content of spleen and thymus indicating its ability to alter metabolic function. We conclude that prolactin may regulate immune function in spleen and thymus. Studies are in progress to identify the regulation of specific gene products by prolactin. PMID- 2991163 TI - Mitoxantrone: propensity for free radical formation and lipid peroxidation- implications for cardiotoxicity. AB - Results of comparative studies on stimulation of the rates of cofactor consumption, superoxide generation and hydrogen peroxide production by mitoxantrone (Novantrone; dihydroxyanthracenedione; MXN), ametantrone (AM), doxorubicin (DOX) and daunorubicin (DNR) in the presence of NADPH-cytochrome P 450 reductase, NADH dehydrogenase, or rabbit hepatic microsomes have been reported. MXN and AM were substantially less effective in stimulating the rate of cofactor oxidation, superoxide formation or hydrogen peroxide production relative to the anthracyclines. In the presence of P-450 reductase, the rate of NADPH oxidation or superoxide generation produced by 100 microM MXN or AM was only 15% and 2% respectively of that produced by 100 microM anthracycline. The effects of MXN and AM on lipid peroxidation in hepatic microsomes, cardiac sarcosomes and cardiac mitochondria were determined and compared with those produced by ADM. MXN and AM at 50 microM inhibited the basal rate of NADPH-dependent rabbit liver microsomal lipid peroxidation by 50%; in contrast, DOX enhanced the rate of hepatic microsomal lipid peroxidation by 2- and 2.5-fold at 100 and 200 microM, respectively. Rabbit cardiac sarcosomal NADPH-dependent lipid peroxidation was inhibited completely at 100 microM anthracenedione. NADH-dependent lipid peroxidation in cardiac mitochondria was diminished by 50 microM MXN and AM, whereas 50 microM DOX produced a 2-fold stimulation in lipid peroxidation. The anthracenediones also effectively inhibited DOX-stimulated lipid peroxidation with 50% inhibition occurring at 4 microM (MXN) and 6 microM (AM). Moreover, both MXN and AM potently inhibited iron (100 microM)-stimulated lipid peroxidation in rabbit hepatic microsomes with 80% inhibition produced by 15 microM anthracenedione.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991165 TI - Radon-222 and 222Rn progeny concentrations measured in an energy-efficient house equipped with a heat exchanger. AB - Radon-222 and 222Rn progeny concentrations, barometric pressure and pressure differentials between inside and outside were measured continuously in the basement of a recently constructed energy-efficient house in metropolitan Denver, CO. Although the monitoring equipment was developed primarily for underground mines, it proved to be applicable for house monitoring. Results indicate that for tightly sealed houses, forced-flow transport does not significantly contribute to the 222Rn present even when the pressure within the house is less than the outside pressure by 0.8 Pa (.006 mm Hg). Calculations of 222Rn levels using diffusion as the primary transport mechanism are in agreement with observed data. The diffusion coefficient of 222Rn in the walls and floor surrounding the basement is higher than values previously reported. Ventilation by means of a heat exchanger reduces the 222Rn levels in accordance with measured air exchange rates, regardless of the pressure differential between inside and outside. PMID- 2991166 TI - Comments on 'A comparison of alpha spectroscopy and gross alpha techniques for determining working level in uranium mines and houses'. PMID- 2991167 TI - A case report of carcinoma originating from aberrant breast tissue. PMID- 2991168 TI - Diet during pregnancy in an Asian community in Britain--energy, protein, zinc, copper, fibre and calcium. AB - Dietary intake of 813 pregnant Harrow Asians of mainly Gujarati descent was compared with the intake of 54 pregnant Europeans living in the same area. The diet of the Europeans had more protein and zinc but less fibre and fat that that of the Harrow Asians. The average rate of weight gain and the energy intakes were similar in both groups. Hindu non-vegetarians and Muslim non-vegetarians differed somewhat, because the Hindus, who are more recent converts to meat-eating, have diets with a lactovegetarian basis to which small amounts of meat have been added. Among the Hindu vegetarians, those who ate eggs and cheese had higher protein and zinc intakes, but otherwise Hindu vegetarians can be regarded as a fairly homogeneous group. Although the protein and zinc intakes of the Asians, particularly the vegetarians, were lower than those of the Europeans, there was no evidence of adverse consequences to the mother and fetus. PMID- 2991169 TI - Colonic transit after fibre supplementation in patients with haemorrhoids. AB - Patients with haemorrhoids often complain of disturbed defaecation which might correspond either to too slow or too fast colonic transit. Colonic transit was determined using a new technique as the distribution of radiopaque markers on a film after a daily intake of 20 markers over 10 days. Nineteen out-patients with a history of haemorrhoids participated in the study. Colonic transit was measured before and after 6 weeks on a bran preparation (Fiberform, 10 g daily) or an ispagula bulk preparation (Lunelax, 10 g daily) in random order. The variation in the distribution of pellets in the colon within the groups was greater than during treatment with fibre preparations. In both cases a 'normalization' of the transit occurred with fibre. No difference was found between the effect of the two fibre preparations on colonic transit. Thus, such changes in transit could be anticipated when the dietary fibre of the diet is increased according to modern recommended dietary allowances. PMID- 2991170 TI - The effect of different types of Finnish bread on postprandial glucose response in diabetic patients. AB - The aim of the study was to investigate the effect of the fibre content of the bread and the effect of the state of graining on the postprandial blood glucose response in insulin-dependent and non-insulin-dependent diabetics. The breads were white wheat bread, mixed wholemeal wheat/rye bread, wholemeal rye bread and grained wholemeal rye bread. Finnish wholemeal rye bread induced a slower postprandial blood glucose response than the mixed wholemeal (wheat/rye) bread (P less than or equal to 0.05) and the white wheat bread (P less than or equal to 0.01). Grained wholemeal rye bread resulted in a similar blood glucose response to that from wholemeal rye bread. PMID- 2991171 TI - Hormonal and metabolic responses to breakfast meals in niddm: comparison of white and whole-grain wheat bread and corresponding extruded products. AB - The post-prandial glucose and hormonal responses were followed in nine non insulin-dependent diabetics after four randomized breakfast meals containing mainly wheat products. The effect of conventionally baked breads was compared with extruded crispbread-like products prepared from white or whole-grain wheat flour, respectively. The extruded whole-grain product gave significantly larger areas under the glucose and insulin curves than the corresponding baked bread, and resulted in higher C-peptide, gastric inhibitory polypeptide, and glucagon concentrations at certain time points. The mean incremental areas under the glucose curves were similar after white bread and the two extruded crispbread like products. There were no significant differences between white and whole grain bread. The results indicate that baked whole-grain wheat bread is to be preferred to corresponding extruded products in non-insulin-dependent diabetes mellitus. PMID- 2991172 TI - Changes in food consumption and its nutritional quality when on a gluten-free diet for dermatitis herpetiformis. AB - Dietary intakes of energy and nutrients were calculated from diet history interviews in 30 patients with dermatitis herpetiformis, before and after 18 months on a gluten-free diet. In spite of great changes in the intake of different foods, the mean intake of dietary fibre did not decrease. There was only a small decrease in the intake of iron in women, while the intake in men did not change. Patients with a previous high intake of gluten had indirect evidence of malabsorption. PMID- 2991173 TI - Dietary fibre intakes of individuals with different eating patterns. AB - This study was undertaken to provide evidence of the variation in the diet necessary to incorporate a high fibre content. Food intakes were measured for seven days by 51 subjects who were either omnivores, vegetarians or vegans. Omnivores consumed the least fibre (23 g), vegetarians significantly more (37 g) and vegans the most (47 g). Men had higher intakes (44 g) than women (30.5 g). Vegans ate the most cereal fibre, but high intake of fibre by both vegetarian groups were due to their preference for unrefined foods, particularly bread. Higher intakes of vegetable fibre by both vegetarian groups were due to high consumption of pulses. Total dietary fibre was significantly related to cereal fibre. As the percentage of fibre increased, the percentage of energy derived from protein decreased. Fat contributed less energy (34 per cent) to the vegan than to the omnivore diet (41 per cent). This study showed that high fibre intake can be achieved on an omnivorous diet and the observation that high fibre diets are possible when the energy content is low, is of relevance to slimming regimes. PMID- 2991174 TI - Long-term follow-up of the treatment of essential hypertension with a high-fibre, low-fat and low-sodium dietary regimen. AB - Thirty-two patients who were being treated with drugs for essential hypertension were started on a dietary regimen comprising high-fibre, low-fat and low-sodium composition. The results are reported of 19 patients who attended after 3 months and for a long-term mean follow-up period of 3.9 years. A significant reduction in systolic (P less than 0.01) and diastolic blood pressure (P less than 0.001) was observed at 3 months and again after 3.9 years (P less than 0.05 and less than 0.001 respectively). This was accompanied by significant weight loss at 3.9 years (P less than 0.05), although the loss was not as great as that at 3 months of study. The number of anti-hypertensive tablets taken by the group was significantly reduced both at 3 months by 60 per cent (P less than 0.01) and after 3.9 years by 49 per cent (P less than 0.01). No changes were observed in mean levels of total serum cholesterol and triglyceride levels although the significant rise in mean HDL-cholesterol levels observed at 3 months (P less than 0.01) was maintained in the long term (P less than 0.05). Analysis of two 24-h dietary histories on non-consecutive days after 3.9 years demonstrated close compliance with the intended regimen, particularly in intakes of sodium, fibre and carbohydrate. However, mean fat intake was 8 per cent above that of the intended dietary regimen. We conclude that the hypotensive response, accompanied by reduction in weight and antihypertensive drug therapy, and improvement in cardiovascular risk, may be maintained in the long term on the modified dietary regimen. PMID- 2991175 TI - Alternating radiotherapy and chemotherapy schedules in small cell lung cancer, limited disease. AB - Sixty-three evaluable patients with limited small cell lung carcinoma were entered into two pilot studies alternating 6 cycles of combination chemotherapy (Doxorubicin 40 mg/m2 d 1; VP16213 75 mg/m2 d 1, 2, 3; Cyclophosphamide 300 mg/m2 d 3, 4, 5, 6; and Methotrexate 400 mg/m2 d 2--plus folinic acid rescue--or Cis Platinum 100 mg/m2 d 2) with 3 courses of mediastinal radiotherapy as induction treatment. The first course of radiotherapy started 10 days after the second cycle of chemotherapy; there was a 7 day rest between chemotherapy and radiotherapy courses. This 6 month induction treatment was followed by a maintenance chemotherapy. The total mediastinal radiation dose was increased from 4500 rad in the first study to 5500 rad in the second. Both protocols obtained a complete response (CR) rate of greater than 85% (with fiberoptic bronchoscopy and histological verification). Local control at 2 years was 61% in the first study and 82% in the second. Relapse-free survival at 2 years was 32 and 37%, respectively. Toxicity was acceptable. We conclude that our results justify further clinical research in alternating radiotherapy and chemotherapy schedules. PMID- 2991176 TI - Oat cell carcinoma of esophagus. PMID- 2991177 TI - Evaluation of a combined dexamethasone suppression/ACTH stimulation test in dogs with hyperadrenocorticism. AB - Twenty-one dogs with hyperadrenocorticism were studied. Six dogs had functioning adrenocortical tumors and 15 had pituitary-dependent hyperadrenocorticism. Each dog was evaluated, using endogenous plasma ACTH, ACTH stimulation, dexamethasone screening, dexamethasone suppression, and combined dexamethasone suppression/ACTH stimulation tests. The ACTH stimulation portion of the combined test was less reliable as a screening test in diagnosing hyperadrenocorticism than was the isolated ACTH stimulation test or the dexamethasone screening test. The dexamethasone suppression portion of the combined test was less reliable in distinguishing dogs with adrenocortical tumors from those with pituitary dependent hyperadrenocorticism than was the endogenous ACTH or isolated dexamethasone suppression test. The combined test is not recommended for use. The ACTH stimulation test is the recommended screening test because of its diagnostic reliability and its subsequent importance as a base line in determining success of mitotane therapy. PMID- 2991178 TI - Clinicopathologic findings resembling hypoadrenocorticism in dogs with primary gastrointestinal disease. AB - Nine dogs with primary gastrointestinal disease had clinical and laboratory findings resembling hypoadrenocorticism. The dogs had histories of anorexia, weakness or lethargy, diarrhea, vomiting, and weight loss. Hypothermia, dehydration, and emaciation also were detected on physical examination. Hyponatremia, hyperkalemia, and abnormally low Na/K ratios were found on laboratory evaluation, but results of ACTH-response tests were not compatible with hypoadrenocorticism. The primary diagnoses were trichuriasis and salmonellosis in 2 dogs, trichuriasis in 5 dogs, and perforated duodenal ulcer in 2 dogs. Most dogs responded to medical or surgical treatment of their primary gastrointestinal disease, and the original electrolyte abnormalities resolved. These findings emphasize the importance of the ACTH-response test in the diagnostic evaluation of dogs with clinicopathologic findings similar to those of hypoadrenocorticism. PMID- 2991179 TI - New 5'-nucleotidase inhibitors, melanocidin A and melanocidin B. I. Taxonomy, fermentation, isolation and biological properties. AB - New 5'-nucleotidase inhibitors named melanocidins A and B were produced by Nocardioides sp. 1681 J. They were isolated from fermentation broth by trichloracetic acid extraction, ethanol precipitation and CM-cellulose and DEAE cellulose column chromatography. They inhibited 5'-nucleotidase activity from snake venom but not from rat liver membrane at 200 micrograms/ml. They also show antitumor activity against melanoma B16. PMID- 2991180 TI - New 5'-nucleotidase inhibitors, melanocidin A and melanocidin B. II. Physico chemical properties and structure elucidation. PMID- 2991181 TI - Studies on a novel cytocidal antibiotic, trienomycin A. Taxonomy, fermentation, isolation, and physico-chemical and biological characteristics. AB - A new antibiotic, trienomycin A, has been isolated from the culture broth of Streptomyces sp. No. 83-16. The physico-chemical characteristics of the antibiotic suggested that it belongs to the group of ansamycin antibiotics with a triene moiety in the molecule. The molecular formula of trienomycin A is C36H50N2O7 (MW 622). The antibiotic possesses potent cytocidal activity against HeLa S3 and PLC hepatoma cells at concentrations of 0.1 and 0.01 micrograms/ml (IC50 value) respectively. However, trienomycin A showed no antimicrobial activity against the bacteria, fungi and yeasts examined with the exception of weak activity versus Piricularia oryzae at a concentration of 1,000 micrograms/ml. PMID- 2991182 TI - Mechanism of inactivation of the polyene antibiotic amphotericin B. Evidence for radical formation in the process of autooxidation. AB - Radical formation during autooxidation of the polyene antibiotic amphotericin B was monitored by following the kinetics of decay of the ESR signal of a stable nitroxide. The kinetics were seen to depend both on antibiotic and on nitroxide concentration. The radicals formed were studied by spin trapping. Three preparations--clinical Fungizone, amphotericin B suspended in buffer, and amphotericin B in buffer - 10% dimethyl sulfoxide--yielded spin adducts of different nature and/or concentrations. In the absence of dimethyl sulfoxide, amphotericin B yielded at least two carbon-centered radicals whose spectra indicated restricted mobility. This suggests that the radicals arise from the whole amphotericin B molecule located in molecular aggregates present in the preparations, and not from smaller and possibly more water-soluble fragments of the antibiotic. In the presence of dimethyl sulfoxide the spin adducts derived from secondary carbon radicals originated from this solvent. Their spectra were indicative of fast tumbling. Direct evidence for autooxidation was obtained by measuring oxygen consumption. All processes examined occurred at the same time scale observed for drug inactivation, in agreement with the idea that loss of activity is due to antibiotic autooxidation. PMID- 2991183 TI - Primary culture of ant venom gland cells. AB - Venom from the ant Pseudomyrmex triplarinus reduces the symptoms and swelling of rheumatoid arthritis. The cells that produce the venom were dissected from larval and pupal ants and culture conditions studied. Cell dissociation, with minimal amount of damage, was done with 0.25% trypsin at 4 degrees C with subsequent use of soybean trypsin inhibitor. A new medium was formulated and epidermal growth factor, fibroblast growth factor, insulin, cAMP, cGMP, and isoproterenol were beneficial. The optimum osmotic pressure was a relatively high 500 mOSM. Conditioning the medium with an established insect cell line was essential for long-term cell survival. Under these culture conditions the structural and metabolic integrity of the cells were maintained for up to 12 mo. PMID- 2991184 TI - Effect of parenteral magnesium on pulmonary function, plasma cAMP, and histamine in bronchial asthma. PMID- 2991185 TI - Effects of K+ and Na+ on the proton motive force of respiring Escherichia coli at alkaline pH. AB - The role of K+ and Na+ in the maintenance of the proton motive force (delta p) was studied in Escherichia coli incubated in alkaline media. Cells respiring in Tris buffer (pH 7.8) that contained less than 100 microEq of K+ and Na+ per liter had a normal delta p of about -165 mV. At pH 8.2, however, the delta p was reduced significantly. The decrease in delta p at pH 8.2 was due to a marked decrease in the transmembrane potential (delta psi), while the internal pH remained at 7.5 to 7.7. When KCl or NaCl, but not LiCl or choline chloride, was added to the cells, the delta psi rose to the values seen at an external pH of 7.8. In addition, choline chloride inhibited the enhancement of delta psi by K+. None of the salts had a significant effect on the internal pH. The effects can be attributed to alterations of K+ or Na+ cycling in and out of the cells via the known K+ and Na+ transport systems. PMID- 2991186 TI - Cryptic plasmid of Neisseria gonorrhoeae: complete nucleotide sequence and genetic organization. AB - The naturally occurring cryptic plasmid pJD1 of Neisseria gonorrhoeae is 4,207 base pairs long and is found in about 96% of gonococcal strains. The total probable coding capacity of pJD1 was determined from the complete nucleotide sequence by using computational probes to identify open reading frames with similar codon usage and by screening for the presence of ribosomal binding sites before the start codons. Candidates for promoters and terminators were also found in the sequence. Based on these findings, we propose a model for the genetic organization of the plasmid. The model predicts two transcriptional units, each composed of five compactly spaced genes. A promoter of one of the transcripts was shown to function in Escherichia coli, and the products of three of the five genes in this operon were identified in minicell expression experiments. Of these, the cppA gene encoded a 9-kilodalton protein, and the cppB and cppC genes both coded for 24-kilodalton proteins. No expression of the other transcriptional unit was detected, but two genes in this operon were expressed in minicells when transcribed from an E. coli promoter. The experimental data were consistent with the model. PMID- 2991187 TI - RecA protein acts at the initiation of stable DNA replication in rnh mutants of Escherichia coli K-12. AB - Escherichia coli rnh mutants lacking RNase H activity are capable of recA+ dependent DNA replication in the absence of concomitant protein synthesis (stable DNA replication). In rnh dnaA::Tn10 and rnh delta oriC double mutants in which the dnaA+-dependent initiation of DNA replication at oriC is completely blocked, the recA200 mutation encoding a thermolabile RecA protein renders both colony formation and DNA synthesis of these mutants temperature sensitive. To determine which stage of DNA replication (initiation, elongation, or termination) was blocked, we analyzed populations of these mutant cells incubated at 30 or 42 degrees C in the presence or absence of chloramphenicol (CM) by dual-parameter (DNA-light scatter) flow cytometry. Incubation at 30 degrees C in the presence of CM resulted in cells with a continuum of DNA content up to seven or more chromosome equivalents per cell. The cultures which had been incubated at 42 degrees C in the absence or presence of CM consisted of cells with integral numbers of chromosomes per cell. It is concluded that active RecA protein is required specifically for the initiation of stable DNA replication. PMID- 2991188 TI - Chromosomal-DNA amplification in Bacillus subtilis. AB - Tetracycline-resistant (Tetr) mutants RAD1, RAD2, RAD6, and RAD7 were isolated from Bacillus subtilis BC92 after protoplasting, polyethylene glycol treatment, and regeneration on a medium containing tetracycline. The Tetr phenotype in RAD1, RAD2, and RAD6 was very stable with less than 5% loss of resistance after 30 generations of growth in the absence of selection. Of the four isolates, three contained amplified chromosomal DNA closely associated with the Tetr phenotype. The intensity of restriction fragments present in HindIII and EcoRI digests of chromosomal DNA from RAD1, RAD6, and RAD7 indicated the presence of tandemly duplicated DNA. Disparity in the size and number of amplified fragments suggested that the tandemly duplicated DNA is different in all three isolates. The sizes of the duplicated DNA present in RAD1, RAD6, and RAD7 were estimated to be 10, 19, and 20 kilobases, respectively. No amplified DNA was detected in RAD2. Results of transductional-mapping studies with PBS1 showed that the tetracycline resistance (tet) loci of RAD1, RAD2, and RAD6 all mapped near the origin of chromosomal replication and close to the guaA locus. Amplified DNA characteristic of RAD1 and RAD6 was cotransduced with the tet locus. Cotransfer of amplified DNA with the guaA locus or other nearby loci in the absence of tet was not observed. In every case, loss of Tetr was accompanied by loss of amplified DNA. A possible explanation for the occurrence of the amplified DNA is presented. PMID- 2991189 TI - Photoreactivation of Escherichia coli reverses umuC induction by UV light. AB - UV mutagenesis in Escherichia coli depends on the presence of a premutagenic lesion in DNA and on the induction of the umuCD gene product as part of the SOS response. Using operon fusions between the E. coli lacZ gene and the SOS genes umuC, uvrB, and dinD, we have affirmed the expected role of the cyclobutane pyrimidine dimer in inducing SOS gene transcription. In addition, we found that photoreactivation of cyclobutane pyrimidine dimers reversed umuC induction to the same extent as it reversed mutagenesis. Therefore, the photoreactivability of UV mutagenesis does not itself identify the cyclobutane pyrimidine dimer as the UV mutagenic lesion in E. coli. PMID- 2991191 TI - Characterization of a trithionate reductase system from Desulfovibrio vulgaris. AB - A trithionate reductase system was isolated and purified from extracts of Desulfovibrio vulgaris. This system reduced trithionate to thiosulfate and consisted of two proteins. One was bisulfite reductase, an enzyme that reduces bisulfite to trithionate, and the second component was designated TR-1. Both enzymes were required to reduce trithionate to thiosulfate. Flavodoxin and cytochrome c3 from D. vulgaris were tested for their ability to function as electron carriers during trithionate reduction. When molecular hydrogen was the source of electrons for the reduction, both flavodoxin and cytochrome c3 were required. In contrast, when the pyruvate phosphoroclastic system was the reductant, flavodoxin alone participated as the electron carrier. The results indicate that flavodoxin, but not cytochrome c3, interacted with the trithionate reductase system. The cytochrome in the hydrogenase-linked assay functioned as an electron carrier between hydrogenase and flavodoxin. PMID- 2991190 TI - Structural genes for flagellar hook-associated proteins in Salmonella typhimurium. AB - The flaW, flaU, and flaV genes of Salmonella typhimurium LT2 were cloned into pBR322. These genes were mapped on the cloned DNA fragments by restriction endonuclease analysis and construction of the deletion derivatives. Their gene products were identified, by the minicell method, as proteins whose molecular weights were estimated to be 59,000 for the flaW product, 31,000 for the flaU product, and 48,000 for the flaV product. These values are identical to those of three species of hook-associated proteins (HAPs), namely, HAP1, HAP3, and HAP2. Furthermore, antibodies against HAP1, HAP3, and HAP2 specifically reacted with the gene products of flaW, flaU, and flaV, respectively. Therefore, we concluded that they are structural genes for HAPs. The antibodies against HAP1 and HAP3 also specifically reacted with the gene products of flaS and flaT of Escherichia coli, respectively. This indicates that these gene products are HAPs in E. coli. This result is consistent with the demonstration that flaS and flaT of E. coli are functionally homologous with flaW and flaU of S. typhimurium. PMID- 2991192 TI - Cloning and sequencing of the beta-lactamase I gene of Bacillus cereus 5/B and its expression in Bacillus subtilis. AB - The beta-lactamases of Bacillus cereus have attracted interest because they are secreted efficiently, because multiple enzymes are frequently present, and because their regulation has unusual features. beta-Lactamase I of strain 5/B is produced constitutively at a high level, and the exoenzyme appears to be several thousand daltons larger than the corresponding product of strain 569/H. We have cloned the gene for 5/B beta-lactamase I in Escherichia coli and B. subtilis and have sequenced the structural portion and the regulatory regions. The 5/B enzyme is produced at a low level in E. coli RR1(pRWY200) and remains cellbound. In B. subtilis it is formed in large amounts, and over 90% of it is released into the medium. There is a large degree of homology between the promoter and leader peptide regions of the 5/B and 569/H genes; both utilize UUG as the translation initiation codon (P. S. F. Mezes, R. W. Blacher, and J. O. Lampen, (J. Biol. Chem. 260:1218-1223, 1985). Although there are significant differences in the peptide segment where processing would be expected to occur, the NH2 terminus of the major 5/B product from B. subtilis BD170(pRWY215) is His-44, which is the same as the NH2 terminus of the major 569/H product from B. subtilis BD170(pRWM5). PMID- 2991193 TI - Molecular cloning of the cls gene responsible for cardiolipin synthesis in Escherichia coli and phenotypic consequences of its amplification. AB - The cls gene responsible for cardiolipin synthesis in Escherichia coli K-12 was cloned in a 5-kilobase-pair DNA fragment inserted in a mini-F vector, pML31, and then subcloned into a 2.0-kilobase-pair fragment inserted in pBR322. The initial selection of the gene was accomplished in a cls pss-1 double mutant that had lesions in both cardiolipin and phosphatidylserine synthases and required either the cls or the pss gene product for normal growth at 42 degrees C in a broth medium, NBY, supplemented with 200 mM sucrose. The cloned gene was identified as the cls gene by the recovery and amplification of both cardiolipin and cardiolipin synthase in a cls mutant as well as by the integration of a pBR322 derivative into its genetic locus at 27 min on the chromosome of a polA1 mutant. The maxicell analysis indicated that a protein of molecular weight 46,000 is the gene product. The cls gene is thus most likely the structural gene coding for cardiolipin synthase. Hybrid plasmids of high copy numbers containing the cls gene were growth inhibitory to pss-I mutants under the above selective conditions, whereas they inhibited neither the growth of pss-I mutants at 30 degrees C nor that of pss+ strains at any temperature. Amplification of cardiolipin synthase activity was observed, but was not proportional to the probable gene dosage (the enzyme activity was at most 10 times that in wild-type cells), and cardiolipin synthesis in vivo was at the maximum 1.5 times that in wild-type strains, implying the presence in E. coli cells of a mechanism that avoids cardiolipin overproduction, which is possibly disadvantageous to proper membrane functions. PMID- 2991194 TI - Regulation of phospholipid synthesis in phosphatidylserine synthase-deficient (chol) mutants of Saccharomyces cerevisiae. AB - chol mutants of Saccharomyces cerevisiae are deficient in the synthesis of the phospholipid phosphatidylserine owing to lowered activity of the membrane associated enzyme phosphatidylserine synthase. chol mutants are auxotrophic for ethanolamine or choline and, in the absence of these supplements, cannot synthesize phosphatidylethanolamine or phosphatidylcholine (PC). We exploited these characteristics of the chol mutants to examine the regulation of phospholipid metabolism in S. cerevisiae. Macromolecular synthesis and phospholipid metabolism were examined in chol cells starved for ethanolamine. As expected, when chol mutants were starved for ethanolamine, the rates of synthesis of the phospholipids phosphatidylethanolamine and PC declined rapidly. Surprisingly, however, coupled to the decline in PC biosynthesis was a simultaneous decrease in the overall rate of phospholipid synthesis. In particular, the rate of synthesis of phosphatidylinositol decreased in parallel with the decline in PC biosynthesis. The results obtained suggest that the slowing of PC biosynthesis in ethanolamine-starved chol cells leads to a coordinated decrease in the synthesis of all phospholipids. However, under conditions of ethanolamine deprivation in chol cells, the cytoplasmic enzyme inositol-1-phosphate synthase could not be repressed by exogenous inositol, and the endogenous synthesis of the phospholipid precursor inositol appeared to be elevated. The implications of these findings with respect to the coordinated regulation of phospholipid synthesis are discussed. PMID- 2991195 TI - Molecular cloning and characterization of the recA gene of Pseudomonas aeruginosa PAO. AB - The recA gene of Pseudomonas aeruginosa PAO has been isolated and introduced into Escherichia coli K-12. Resistance to killing by UV irradiation was restored in several RecA-E. coli K-12 hosts by the P. aeruginosa gene, as was resistance to methyl methanesulfonate. Recombination proficiency was also restored, as measured by HfrH-mediated conjugation and by the ability to propagate Fec-phage lambda derivatives. The cloned P. aeruginosa recA gene restored both spontaneous and mitomycin C-stimulated induction of lambda prophage in lysogens of a recA strain of E. coli K-12. PMID- 2991196 TI - Determination of DNA sequences containing methylcytosine in Bacillus subtilis Marburg. AB - The methylcytosine-containing sequences in the DNA of Bacillus subtilis 168 Marburg (restriction-modification type BsuM) were determined by three different methods: (i) examination of in vivo-methylated DNA by restriction enzyme digestion and, whenever possible, analysis for methylcytosine at the 5' end; (ii) methylation in vitro of unmethylated DNA with B. subtilis DNA methyltransferase and determination of the methylated sites; and (iii) the methylatability of unmethylated DNA by B. subtilis methyltransferase after potential sites have been destroyed by digestion with restriction endonucleases. The results obtained by these methods, taken together, show that methylcytosine was present only within the sequence 5'-TCGA-3'. The presence of methylcytosine at the 5' end of the DNA fragments generated by restriction endonuclease AsuII digestion and the fact that in vivo-methylated DNA could not be digested by the enzyme XhoI showed that the recognition sequences of these two enzymes contained methylcytosine. As these two enzymes recognized a similar sequence containing a 5' pyrimidine (Py) and a 3' purine (Pu), 5'-PyTCGAPu-3', the possibility that methylcytosine is present in the complementary sequences 5'-TTCGAG-3' and 5'-CTCGAA-3' was postulated. This was verified by the methylation in vitro, with B. subtilis enzyme, of a 2.6 kilobase fragment of lambda DNA containing two such sites and devoid of AsuII or XhoI recognition sequences. By analyzing the methylatable sites, it was found that in one of the two PyTCGAPu sequences, cytosine was methylated in vitro in both DNA strands. It is concluded that the sequence 5'-PyTCGAPu-3' is methylated by the DNA methyltransferase (of cytosine) of B. subtilis Marburg. PMID- 2991197 TI - Genetic analysis of the cholera toxin-positive regulatory gene toxR. AB - Southern blot analysis with a toxR-specific gene probe indicates that Vibrio cholerae 569B has a 1.2-kilobase deletion near the toxR gene. Heterologous conjugative crosses were carried out between the EI Tor strain RV79 and 569B tox mutants. Tox+ recombinants showed the same linkage properties to the his locus as to the previously mapped tox locus of 569B. Southern blot analysis with the toxR probe of the Tox+ recombinants obtained in these heterologous crosses showed that these recombinants had replaced the V. cholerae 569B (recipient) toxR DNA with the V. cholerae RV79 (donor) toxR DNA, indicating that tox and toxR are the same locus. However, the Tox+ recombinants synthesized an amount of toxin intermediate between the level observed for wild-type RV79 and 569B strains, suggesting there is a difference in the ability of toxR genes from different strains to activate ctx. About half of the mutations which suppress the phenotype of hypertoxinogenic locus htx are unlinked to htx and in addition have a hypotoxinogenic phenotype relative to that of the wild type. Most of these hypotoxinogenic, second-site suppressors show a linkage to his similar to the linkage of toxR to his and are therefore probably mutations in toxR. These results indicate that the toxR gene product is required for ctx expression and that a functional toxR gene is required for the effect of an htx mutation to be seen. PMID- 2991198 TI - Genetics of methyl-accepting chemotaxis proteins in Escherichia coli: null phenotypes of the tar and tap genes. AB - The tar and tap genes are located adjacent to one another in an operon of chemotaxis-related functions. They encode methyl-accepting chemotaxis proteins implicated in tactic responses to aspartate and maltose stimuli. The functional roles of these two gene products were investigated by isolating and characterizing nonpolar, single-gene deletion mutants at each locus. Deletions were obtained by selecting for loss or a defective Mu d1 prophage inserted in either the tar or tap gene. The extent of the tar deletions was determined by genetic mapping with Southern hybridization. Representative deletion mutants were surveyed for chemotactic responses on semisolid agar and by temporal stimulation in a tethered cell assay to assess flagellar rotational responses to chemoeffector compounds. The tar deletion strains exhibited complete loss of aspartate and maltose responses, whereas the tap deletion strains displayed a wild-type phenotype under all conditions tested. These findings indicate that the tap function is unable to promote chemotactic responses to aspartate and maltose, and its role in chemotaxis remains unclear. PMID- 2991199 TI - Mutations that affect utilization of a promoter in stationary-phase Bacillus subtilis. AB - Transcription of the ctc gene in Bacillus subtilis is activated only after exponentially growing cells enter stationary phase. The promoter of the ctc gene is utilized in vitro by two minor forms of RNA polymerase, E sigma 37 and E sigma 32, but not by the most abundant form of RNA polymerase, E sigma 55. We have used the ctc promoter to direct transcription of the xylE gene on plasmid pLC4 and observed that xylE was expressed only in stationary-phase B. subtilis. We also have constructed a series of homologous plasmids that differ only by specific base substitutions in the ctc promoter. We observed that the base substitutions that affected utilization of the ctc promoter in vivo (xylE expression) were the same as those that we had previously shown to affect utilization of the promoter in vitro by E sigma 37 and E sigma 32. We conclude that it is likely that the ctc promoter is utilized in vivo by E sigma 37 or E sigma 32. PMID- 2991200 TI - Bacteroides intermedius binds fibrinogen. AB - The binding of Bacteroides intermedius VPI 8944 to human fibrinogen has been characterized. The binding is time dependent, at least partially reversible, saturable, and specific. On an average, a maximum of 3,500 fibrinogen molecules bind per bacterial cell, with a dissociation constant of 1.7 X 10(-11) M. These bacteria also exhibit a fibrinogenolytic activity which can be partially inhibited by protease inhibitors. Bacteria release fibrinogenolytic activity into the surrounding medium without loss of binding activity, but more pronounced fibrinogen breakdown occurs when 125I-labeled fibrinogen is associated with the bacteria, suggesting that fibrinogen is degraded at the cell surface. Fibrinogen binding by B. intermedius might represent a mechanism of bacterial tissue adherence. PMID- 2991201 TI - Purification and characterization of selenocysteine beta-lyase from Citrobacter freundii. AB - The purification and characterization of bacterial selenocysteine beta-lyase, an enzyme which specifically catalyzes the cleavage of L-selenocysteine to L-alanine and Se0, are presented. The enzyme, purified to near homogeneity from Citrobacter freundii, is monomeric with a molecular weight of ca. 64,000 and contains 1 mol of pyridoxal 5'-phosphate as a cofactor per mol of enzyme. L-Selenocysteine is the sole substrate (Km, 0.95 mM). L-Cysteine is a competitive inhibitor of the enzyme (Ki, 0.65 mM). The enzyme also catalyzes the alpha, beta elimination of beta-chloro-L-alanine to form NH3, pyruvate, and Cl- and is irreversibly inactivated during the reaction. The physicochemical properties, e.g., amino acid composition and subunit structure, of the bacterial enzyme are fairly different from those of the pig liver enzyme (Esaki et al., J. Biol. Chem. 257:4386-4391, 1982). However, the catalytic properties of both enzymes, e.g., substrate specificity and inactivation by the substrate or a mechanism-based inactivator, beta-chloro-L-alanine, are very similar. PMID- 2991202 TI - Partial purification and characterization of glutaryl-coenzyme A dehydrogenase, electron transfer flavoprotein, and electron transfer flavoprotein-Q oxidoreductase from Paracoccus denitrificans. AB - Glutaryl-coenzyme A (CoA) dehydrogenase and the electron transfer flavoprotein (ETF) of Paracoccus denitrificans were purified to homogeneity from cells grown with glutaric acid as the carbon source. Glutaryl-CoA dehydrogenase had a molecular weight of 180,000 and was made up of four identical subunits with molecular weights of about 43,000 each of which contained one flavin adenine dinucleotide molecule. The enzyme catalyzed an oxidative decarboxylation of glutaryl-CoA to crotonyl-CoA, was maximally stable at pH 5.0, and lost activity readily at pH values above 7.0. The enzyme had a pH optimum in the range of 8.0 to 8.5, a catalytic center activity of about 960 min-1, and apparent Michaelis constants for glutaryl-CoA and pig liver ETF of about 1.2 and 2.5 microM, respectively. P. denitrificans ETF had a visible spectrum identical to that of pig liver ETF and was made up of two subunits, only one of which contained a flavin adenine dinucleotide molecule. The isoelectric point of P. denitrificans ETF was 4.45 compared with 6.8 for pig liver ETF. P. denitrificans ETF accepted electrons not only from P. denitrificans glutaryl-CoA dehydrogenase, but also from the pig liver butyryl-CoA and octanoyl-CoA dehydrogenases. The apparent Vmax was of similar magnitude with either pig liver or P. denitrificans ETF as an electron acceptor for these dehydrogenases. P. denitrificans glutaryl-CoA dehydrogenase and ETF were used to assay for the reduction of ubiquinone 1 by ETF Q oxidoreductase in cholate extracts of P. denitrificans membranes. The ETF-Q oxidoreductase from P. denitrificans could accept electrons from either the bacterial or the pig liver ETF. In either case, the apparent Km for ETF was infinitely high. P. denitrificans ETF-Q oxidoreductase was purified from contaminating paramagnets, and the resultant preparation had electron paramagnetic resonance signals at 2.081, 1.938, and 1.879 G, similar to those of the mitochondrial enzyme. PMID- 2991203 TI - Cloning of GLN4: an essential gene that encodes glutaminyl-tRNA synthetase in Saccharomyces cerevisiae. AB - The structural gene for glutaminyl-tRNA synthetase has been isolated from a gene bank of Saccharomyces cerevisiae chromosomal DNA. Cloning was achieved by complementation of a recently described yeast strain that is auxotrophic for glutamine. A multicopy recombinant plasmid with a 5-kilobase-pair genomic insert conferred sixfold elevation in glutaminyl-tRNA synthetase activity and restored a Gln+ phenotype to strains that were Gln- by virtue of a mutant gln4 allele. Subfragments of the 5-kilobase insert directed integration of URA3 to GLN4. Further experiments established that GLN4 is an essential gene that is located on chromosome XV. RNA blots with a GLN4-specific probe detected a single transcript of approximately 2,900 nucleotides. PMID- 2991204 TI - Cloning and location of the dgsA gene of Escherichia coli. AB - The dgsA locus of Escherichia coli was isolated on plasmids obtained from the library of L. Clarke and J. Carbon (Cell 9:91-99, 1976). Restriction fragment analysis and further subcloning demonstrated that the gene is located at kilobase 425 on the Bouche physical map of the terminus region (J. P. Bouche, J. Mol. Biol., 154:1-20, 1982). This corresponds to 35.2 min on the Bachmann genetic map (B. J. Bachmann, Microbiol. Rev. 47:180-230, 1983). PMID- 2991206 TI - Mapping of a gene that regulates hemolysin production in Vibrio cholerae. AB - A gene that regulates the hemolysin structural gene (hly) was found to be tightly linked to the tox-1000 locus of Vibrio cholerae RJ1 and separated from hly by a large section of the V. cholerae genetic map. This hemolysin regulatory gene was designated hlyR. PMID- 2991205 TI - Sequence of the Escherichia coli pheST operon and identification of the himA gene. AB - The complete nucleotide sequence of the Escherichia coli pheST operon coding for the two subunits of phenylalanyl-tRNA synthetase (an alpha 2 beta 2-type enzyme) has been determined. Another open reading frame (prp) was revealed downstream from pheT which was identified as himA, the gene for the alpha subunit of the integration host factor. PMID- 2991207 TI - Phosphoinositide breakdown as an indirect link between stimulation and aggregation of rat platelets by thrombin and collagen. AB - When washed rat platelets (1.5 x 10(9)/ml) were stimulated by a threshold concentration of thrombin (0.3 unit/ml) or collagen (10 micrograms/ml), a lag period of about 10 or 30 s, respectively, was seen before the start of aggregation. During the lag period, [32P]phosphatidylinositol 4,5-bisphosphate was degraded as the earliest event within 5-10 s of addition of the stimulus. However, though the extent of phosphatidylinositol 4,5-bisphosphate degradation within 10 s of addition of collagen was greater than that within 20 s of addition of thrombin (0.3 unit/ml), a lag of about 20 s remained before the initiation of aggregation by collagen. This casts doubt on the hypothesis that the stimulus dependent phosphatidylinositol 4,5-bisphosphate breakdown induces the aggregation of platelets. Phosphatidylinositol labeled with 32Pi or [1-14C]arachidonic acid was scarcely degraded during the lag period. As aggregation proceeded, [14C arachidonic acid]phosphatidylinositol was degraded with generation of diacylglycerol, phosphatidic acid, arachidonic acid and its metabolites. The maximum aggregation by collagen of rat platelets in which arachidonic acid of phospholipids was replaced in vivo with eicosapentaenoic acid was reduced, but that by thrombin was not, though reduction of thromboxane A2 generation was caused by both stimuli. Indomethacin also fully inhibited the aggregation induced by collagen, but not that induced by thrombin. Hence, thromboxane A2 is required for full aggregation by collagen, but not that by thrombin. These results indicate that thrombin-induced phosphoinositide metabolism may proceed independently of aggregation. PMID- 2991208 TI - Partial purification and characterization of type I DNA topoisomerase from Bacillus stearothermophilus. AB - Type I DNA topoisomerase was partially purified from Bacillus stearothermophilus by ammonium sulfate precipitation and column chromatographies on phosphocellulose, DEAE-cellulose and heparin-agarose. On heparin-agarose chromatography, topoisomerase I activity was separated into three fractions (designated Fractions A, B, and C). Each fraction was further subjected to gel filtration on Sephacryl S-200. From electrophoretic analysis on polyacrylamide gel, Fraction A was found to contain two enzyme species having molecular weights of 110,000 and 100,000, and Fraction B one enzyme species with a molecular weight of 80,000. The molecular weight of the enzyme in Fraction C was estimated to be around 150,000 by gel filtration. The enzymes in Fractions A and B exhibited little activity in the presence of Mg2+, while the activity was increased remarkably by NaCl with Mg2+. No activity was observed in the presence of NaCl alone. The enzyme in Fraction C required only Mg2+ for full activity. With Fraction A, the topoisomerase I-induced cleavage sites on tetracycline-resistant plasmid pNS1 (2.55 megadaltons) were mapped. Fraction A cleaved the DNA at ten specific sites. These sites were compared to those of the Haemophilus gallinarum enzyme, which have already been mapped (Shishido et al. (1983) Biochem. Biophys. Acta 740, 108). The results showed that there is a remarkably coincidence between the cleavage sites induced by the B. stearothermophilus and H. gallinarum enzymes. PMID- 2991209 TI - Trinitrophenylation of rabbit skeletal myosin by 2,4,6-trinitrobenzene sulfonate and treatment of trinitrophenyl myosin with dithiothreitol. AB - Rabbit skeletal myosin was trinitrophenylated with 2,4,6-trinitrobenzene sulfonate (TNBS) in the presence or absence of inorganic pyrophosphate (PP1). When myosin trinitrophenylated either in the presence or absence of PP1 was treated with dithiothreitol (DTT), the absorbance at 345 nm of both trinitrophenylated myosins was decreased, as though the trinitrophenyl groups bound to myosin were removed. The DTT treatment also essentially reversed the inhibition of the EDTA-ATPase and Ca-ATPase activities that was caused by trinitrophenylation of myosin. These effects of trinitrophenylation and of DTT treatment were independent of the presence or absence of PP1 during the trinitrophenylation. In contrast, the PP1-induced formation of a difference spectrum of trinitrophenylated myosin was not affected by the DTT treatment. On the basis of these observations, it is suggested that the "reactive lysine residues," trinitrophenylation of which resulted in inhibition of the ATPase activities, are different from those whose trinitrophenyl groups show an altered spectrum on addition of PP1. PMID- 2991210 TI - Ectoenzyme release from rat liver and kidney by phosphatidylinositol-specific phospholipase C. AB - Ectoenzyme release from rat liver and kidney by phosphatidylinositol (PI) specific phospholipase C of Bacillus thuringiensis was studied. Alkaline phosphatase and 5'-nucleotidase were released from rat kidney slices to extents of up to 60% and 30%, respectively. Release of alkaline phosphatase was observed at lower amounts of PI-specific phospholipase C than that of 5'-nucleotidase. Both enzymes were more easily released from microsomal fractions or free cells. From kidney cells, alkaline phosphatase was released without cell lysis, and more than 80% release of alkaline phosphatase was observed at 3.8% hydrolysis of PI. Isoelectric focusing profiles of alkaline phosphatase released by PI-specific phospholipase C were significantly different from the control in the cases of both rat liver and kidney. Lubrol-solubilized alkaline phosphatase was eluted at the void volume of a Toyopearl HW-55 column, while the enzyme obtained by further treatment with PI-specific phospholipase C was eluted in the lower-molecular weight region corresponding to 100,000-110,000 daltons. Furthermore, Lubrol solubilized phosphatase became more thermostable on treatment with PI-specific phospholipase C. PMID- 2991211 TI - A spin-label study on human high density lipoprotein. AB - Human plasma high density lipoproteins (HDL) have been labeled with N-(1-oxyl 2,2,6,6-tetramethyl-4-piperidinyl)maleimide (NEM-TEMPO). The spin-labeled HDL exhibited an ESR spectrum containing signals of both strongly immobilized and weakly immobilized components by the reaction with a high concentration of NEM TEMPO, while an ESR spectrum containing only signals of a strongly immobilized component range between 4 degrees C and 37 degrees C, the signal height of the strongly immobilized component exhibited reversible temperature-dependent changes, whereas that of the weakly immobilized component changed irreversibly at temperatures above 25 degrees C. The activation energy of the irreversible change was estimated to be 26 kcal per mol. The strongly immobilized component was derived from NEM-TEMPO which modified apolipoprotein A-I covalently, while the weakly immobilized component was derived from NEM-TEMPO noncovalently bound to HDL. The rate of binding of NEM-TEMPO to either the strongly binding or weakly binding sites and the number of the strongly binding sites in apolipoprotein A-I were estimated to be 125 M-1.day-1 and 1.78, respectively. The binding of NEM TEMPO to the strongly binding sites was suppressed greatly by pretreatment of HDL with 2,4,6-trinitrobenzene sulfonic acid (TNBS). The slow reaction and suppression with TNBS suggest that NEM-TEMPO binds to some amino acid residue, probably a lysine residue, in apoprotein A-I. The strongly immobilized and weakly immobilized components were reduced almost completely by ascorbate at the same rate, 0.048 min-1 at pH 7.4 and at 4 degrees C. PMID- 2991212 TI - Complete amino acid sequence of cytochrome c from the honeybee, Apis mellifera, and evolutionary relationship of the honeybee to other insects on the basis of the amino acid sequence. AB - The complete amino acid sequence of cytochrome c purified from the honeybee, Apis mellifera was determined. Only one molecular species of cytochrome c was found in the honeybee throughout its metamorphic stages. On the basis of a comparison of the amino acid sequence of honeybee cytochrome c with those of cytochromes c from other insects, it seems that the bee has evolutionarily appeared earlier than would be expected from the morphological and fossil evidence. If the classical phylogenetic relationships of the honeybee are correct, the evolutionary rate of cytochrome c must have been more rapid in the honeybee than in other insects. PMID- 2991213 TI - Two different preparations of subfragment-1 from chicken skeletal myosin and from porcine cardiac myosin. AB - Two different subfragment-1 preparations were obtained from either skeletal or cardiac myosin. They were identical in the heavy chain and light chain compositions but different in the pH dependence of the Ca-ATPase activity and in the relationship with "reactive lysine residues" (RLR). PMID- 2991214 TI - Cytoplasmic 5'-nucleotidase catalyzes acyclovir phosphorylation. AB - A cytoplasmic 5'-nucleotidase (EC 3.1.3.5) can catalyze the phosphorylation of inosine (Worku, Y., and Newby, A.C. (1982) Biochem. J. 205, 503-510). This enzyme was purified to determine whether it could catalyze the formation of trace levels of phosphorylated acyclovir (ACV), a nucleoside analog with antiherpes activity. Acyclovir phosphorylating activity from rat liver co-chromatographed with the enzyme throughout the 1200-fold purification and through size exclusion chromatography or polyacrylamide-gel electrophoresis. In addition, the pH optimum, ATP stimulation, and phosphate inhibition of the ACV phosphorylating activity paralleled those of the 5'-nucleotidase. Finally, ACV phosphorylation was competitively inhibited by inosine (Kis = 6.5 mM; K'm (inosine) = 5.0 mM). This was consistent with phosphorylation at a common catalytic site. In addition to inosine and ACV, the guanine derivatives Guo, dGuo, 9-beta-D arabinofuranosylguanine, and 9-(1,3-dihydroxy-2-propoxymethyl)guanine were substrates for the enzyme. The relative phosphorylation rates were, respectively, 100, 0.7, 19, 4, 0.3, and 0.7, at 0.1 mM phosphate acceptor. Approximate K'm values were, respectively, 5, 90, 10, 10, greater than 100, and greater than 100 mM. Although the substrate activity of ACV with the 5'-nucleotidase was inefficient, it appeared to be sufficient to account for the small amounts of ACV phosphates formed in uninfected cells. PMID- 2991215 TI - Herpes simplex type 1 ribonucleotide reductase. Mechanism studies with inhibitors. AB - Several known inhibitors of mammalian ribonucleotide reductase were studied for their interactions with herpes simplex virus type 1 (HSV-1) ribonucleotide reductase. MAIQ (4-methyl-5-amino-1-formylisoquinoline thiosemicarbazone) produced apparent inactivation of HSV-1 ribonucleotide reductase. Only catalytically cycling, not resting, enzyme could be inactivated. Double reciprocal replots of the rates of inactivation versus the concentration of MAIQ indicated that a reversible complex with the enzyme was formed prior to inactivation. In the presence of 10 microM CDP, the maximum rate of inactivation was 20 per h (t1/2 = 3 min). The half-maximum rate was achieved at about 15 microM MAIQ. INOX (periodate-oxidized inosine) also appeared to inactivate HSV-1 ribonucleotide reductase. In contrast to MAIQ, it readily inactivated resting as well as cycling enzyme. CDP retarded the rates of inactivation by INOX. An initial reversible complex between INOX and enzyme was not detectable under the conditions used. IMPY (2,3-dihydro-1H-pyrazolo(2,3-a)imidazole) and guanazole (3,5-diamino-1,2,4-triazole) produced reversible inhibition. Although the data with both inhibitors were most consistent with the noncompetitive inhibition model (versus CDP), the data with guanazole were also marginally consistent with the uncompetitive model. PMID- 2991216 TI - cAMP-dependent protein kinase activation lowers hepatocyte cAMP. AB - Rat hepatocyte protein kinase was activated by incubating the cells with various cAMP analogs. Boiled extracts were then prepared and Sephadex G-25 chromatography was carried out. The G-25 procedure separated the analogs from cAMP since the resin had the unexpected property of binding cyclic nucleotides with differing affinities. Separation was necessary because the analogs would otherwise interfere with the sensitive protein kinase activation method developed for assay of cAMP. The cAMP analogs, but not 5'-AMP, lowered basal cAMP by 50-70%. The effect was rapid, analog concentration-dependent, and occurred parallel with phosphorylase activation, suggesting that the cAMP analogs act through cAMP dependent protein kinase activation. A cAMP analog completely blocked the cAMP elevation produced by relatively low concentrations of glucagon, but did not block the phosphorylase response, indicating that the cAMP analog substitutes for cAMP as the intracellular activator of protein kinase. One implication of the results is that elevation of cAMP and protein kinase activity by hormones has a negative feedback effect on the cellular cAMP level. PMID- 2991218 TI - Myosin light chain phosphatase. Effect on the activation and relaxation of gizzard smooth muscle skinned fibers. AB - Skinned cells of chicken gizzard were used to study the effect of a smooth muscle phosphatase (SMP-IV) on activation and relaxation of tension. SMP-IV has previously been shown to dephosphorylate light chains on myosin. When this phosphatase was added to submaximally Ca2+-activated skinned cells, tension increased while phosphorylation of myosin light chains decreased. In contrast, when the myosin phosphatase was added to cell bundles activated in the absence of Ca2+ by a Ca2+-insensitive myosin light chain kinase, tension and phosphorylation of the myosin light chains both decreased. These data suggest that Ca2+ inhibits the deactivation of tension even when myosin light chains are dephosphorylated to a low level. Furthermore, comparison of Ca2+-activated cells caused to relax in CTP, in the presence or absence of Ca2+, shows that cells in the presence of Ca2+ do not relax completely, whereas in the absence of Ca2+ cells completely relax. Solutions containing Ca2+ and CTP, however, are incapable of generating tension from the resting state. Endogenous myosin light chain kinase is not active in solutions containing CTP and dephosphorylation of myosin light chains occurs in CTP solutions both in the presence and absence of Ca2+. These data imply that Ca2+ inhibits relaxation even though myosin light chains are dephosphorylated. These data are consistent with a model wherein an obligatory Ca2+-activated myosin light chain phosphorylation is followed by a second Ca2+ activation process for further tension development or maintenance. PMID- 2991217 TI - Degradation without apparent change in size of molybdate-stabilized nonactivated glucocorticoid-receptor complexes in rat thymus cytosol. AB - We have investigated the stability of the [3H]dexamethasone 21-mesylate-labeled nonactivated glucocorticoid-receptor complex in rat thymus cytosol containing 20 mM sodium molybdate. Cytosol complexes were analyzed under nondenaturing conditions by gel filtration chromatography in the presence of molybdate and under denaturing conditions by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. When analyzed under nondenaturing conditions, complexes from fresh cytosol and from cytosol left for 2 h at 3 degrees C eluted from gel filtration as a single peak of radioactivity with a Stokes radius of approximately 7.7 nm, suggesting that no proteolysis of the complexes had occurred in either cytosol. When analyzed under denaturing conditions, however, whereas the fresh cytosol gave a receptor band on sodium dodecyl sulfate polyacrylamide gel electrophoresis at Mr approximately 90,000 (corresponding to the intact complex), the cytosol that had been left for 2 h at 3 degrees C gave only a fragment (Mr approximately 50,000). This fragment, just as the intact complex, could be thermally activated to a DNA-binding form. Proteolysis of the receptor could be blocked by preparing the cytosol in the presence of EGTA, leupeptin, or a heat-stable factor present in the cytosol of rat liver and WEHI-7 mouse thymoma cells. From these results we conclude: (i) 20 mM molybdate does not protect the nonactivated glucocorticoid-receptor complex present in rat thymus cytosol against proteolysis under conditions which are commonly used for cell free labeling of the receptor, and (ii) the demonstration of a Stokes radius of approximately 8 nm for the nonactivated glucocorticoid-receptor complex is not sufficient to indicate that the receptor complex is present in its intact form. PMID- 2991219 TI - Interaction of crotoxin and its isolated subunits with spin-labeled fatty acids. AB - We have investigated the interaction of crotoxin (component A-component B complex) and of its isolated phospholipase subunit (component B) with hydrophobic compounds by ESR, using spin-labeled fatty acids as probes. The phospholipase subunit alone (component B) binds more than three labeled fatty acid molecules/molecule with different affinities, the highest corresponding to a Kd of 10 microM in the case of 5-doxyl palmitic acid. In contrast, the noncatalytic subunit (component A) and the crotoxin complex do not bind fatty acids. ESR studies of the component B-fatty acid complex reveal a strong immobilization of the whole length of the fatty acid chain, strong spin-spin interactions between bound fatty acids, and nonaccessibility of the bound paramagnetic probe to Ni2+ ions. This suggests that the phospholipase component B possesses a hydrophobic cleft which may contain one or two fatty acids. This hydrophobic cleft is not accessible to spin-labeled fatty acids in the crotoxin complex. An overall rotational correlation time of about 200 ns of the phospholipase component B was determined by saturation transfer ESR. This high value is incompatible with the diffusion of a polypeptide of 14,500 molecular weight. The hydrodynamic analysis of the fatty acid-component B complex led us to estimate an apparent molecular weight of 95,000 which reveals that fatty acids induce the formation of polymers (most probably octamers) of component B. We propose a model in which the phospholipase component B exists in two conformational states which differ by their hydrophobicity. PMID- 2991221 TI - Separation and characterization of a phosphatidylinositol kinase activity that co purifies with the epidermal growth factor receptor. AB - Two retroviral protein-tyrosine kinases, v-src and v-ros, have been reported to possess phosphatidylinositol (PtdIns) kinase activity. Because the epidermal growth factor (EGF) receptor is a protein-tyrosine kinase with structural homology to p60v-src and because EGF stimulates PtdIns turnover in A431 cells, the EGF receptor has been examined for PtdIns kinase activity. Preparations of the EGF receptor, isolated from A431 cells and purified by two different methods of affinity chromatography, possessed an associated PtdIns kinase activity. This activity which co-purified with the EGF receptor represented only about 2% of the total PtdIns kinase activity of A431 membranes, and there was no correlation between the number of EGF receptors and the amount of PtdIns kinase activity in membranes from various cell types. A peptide substrate, angiotensin II, and PtdIns did not compete with each other as substrates for the protein-tyrosine and PtdIns kinase activities of the EGF receptor. When self-phosphorylated EGF receptor was fractionated by Sephacryl S-300 gel permeation chromatography, the peak of PtdIns kinase activity was separated from the comigrating peak of protein tyrosine kinase activity and the self-phosphorylated EGF receptor. These results indicate that the protein-tyrosine kinase and PtdIns kinase activities which co purify with the EGF receptor reside on different molecules. Angiotensin II and PtdIns did not compete as substrates for p60v-src isolated by immunoabsorption with a monoclonal antibody, suggesting that PtdIns kinase activity may also not be intrinsic to p60v-src. PMID- 2991220 TI - Receptor-mediated uptake of remnant lipoproteins by cholesterol-loaded human monocyte-macrophages. AB - Normal human monocyte-macrophages were cholesterol-loaded, and the rates of uptake and degradation of several lipoproteins were measured and compared to rates in control cells. Receptor activities for 125I-rabbit beta-very low density lipoproteins (beta-VLDL), 125I-human low density lipoprotein, and 125I-human chylomicrons were down-regulated in cholesterol-loaded cells; however, the rate of uptake and degradation of 125I-human chylomicron remnants was unchanged from control cells. Cholesterol-loaded alveolar macrophages from a Watanabe heritable hyperlipidemic rabbit, which lack low density lipoprotein receptors, showed receptor down-regulation for 125I-beta-VLDL but not for 125I-human chylomicron remnants. In addition to chylomicron remnants, apo-E-phospholipid complexes competed for 125I-chylomicron remnant uptake, but apo-A-I-phospholipid complexes did not. Chylomicrons competed for lipoprotein uptake in control cells but were not recognized under conditions of cholesterol loading. Chylomicron remnants and beta-VLDL were equally effective in competing for 125I-beta-VLDL and 125I chylomicron remnant uptake in cholesterol-loaded macrophages. When normal human monocyte-macrophages were incubated in serum supplemented with chylomicron remnants, the cholesteryl ester content increased 4-fold over cells incubated in serum with low density lipoprotein added. We conclude: 1) specific lipoprotein receptor activity persists in cholesterol-loaded cells; 2) this receptor activity recognizes lipo-proteins (at least in part) by their apo-E content; and 3) cholesteryl ester accumulation can occur in monocyte-macrophages incubated with chylomicron remnants. PMID- 2991222 TI - Interaction of Dictyostelium discoideum lysosomal enzymes with the mammalian phosphomannosyl receptor. The importance of oligosaccharides which contain phosphodiesters. AB - Mammalian cell lysosomal enzymes or phosphorylated oligosaccharides derived from them are endocytosed by a phosphomannosyl receptor (PMR) found on the surface of fibroblasts. Various studies suggest that 2 residues of Man-6-P in phosphomonoester linkage but not diester linkage (PDE) are essential for a high rate of uptake. The lysosomal enzymes of the slime mold Dictyostelium discoideum are also recognized by the PMR on these cells; however, none of the oligosaccharides from these enzymes contain 2 phosphomonoesters. Instead, most contain multiple sulfate esters and 2 residues of Man-6-P in an unusual PDE linkage. In this study I have tried to account for the unexpected highly efficient uptake of the slime mold enzymes. The results show that nearly all of the alpha-mannosidase molecules contain the oligosaccharides required for uptake, and that each tetrameric, holoenzyme molecule has sufficient carbohydrate for an average of 10 Man8GlcNAc2 oligosaccharides. None of the oligosaccharides or glycopeptides from the lysosomal enzymes bind to an immobilized PMR, but those with 2 PDE show slight interaction. Competition of 125I-beta-glucosidase uptake by various carbohydrate-containing fractions indicates that the best inhibitors are those with 2 PDE, either with or without sulfate esters. Furthermore, the uptake of a lysosomal enzyme isolated from a mutant strain (modA), which produces oligosaccharides with only 1 but not 2 PDE, is about 10-fold less than the uptake of wild-type enzyme which has predominantly 2 PDE. Complete denaturation of 125I labeled wild-type beta-glucosidase in sodium dodecyl sulfate/dithiothreitol also reduces its uptake by about 10-fold. Taken together, these results suggest that the interactions of multiple, weakly binding oligosaccharides, especially those with 2 PDE, are important for the high rate of uptake of the slime mold enzymes. The conformation of the protein may be important in orienting the oligosaccharides in a favorable position for binding to the PMR. PMID- 2991223 TI - Molecular alteration of a muscarinic acetylcholine receptor system during synaptogenesis. AB - Biochemical properties of the muscarinic acetylcholine receptor system of the avian retina were found to change during the period when synapses form in ovo. Comparison of ligand binding to membranes obtained before and after synaptogenesis showed a significant increase in the affinity, but not proportion, of the high affinity agonist-binding state. There was no change in receptor sensitivity to antagonists during this period. Pirenzepine binding, which can discriminate muscarinic receptor subtypes, showed the presence of a single population of low affinity sites (M2) before and after synaptogenesis. The change in agonist binding was not due to the late development of receptor function; tests for receptor-stimulated phosphatidylinositol turnover and for modulation of agonist binding by guanylylimidodiphosphate showed functional coupling to be present several days prior to the onset of synapse formation. However, detergent solubilization of membranes eliminated differences in agonist binding between receptors from embryos and hatched chicks, suggesting a developmental change in interactions of the receptor with functionally related membrane components. A possible basis for altered interactions was obtained from isoelectric point data showing that the muscarinic receptor population underwent a transition from a predominantly low pI form (4.25) in 13 day embryos to a predominantly high pI form (4.50) in newly hatched chicks. The possibility that biochemical changes in the muscarinic receptor play a role in differentiation of the system by controlling receptor position on the surface of nerve cells is discussed. PMID- 2991224 TI - Identification of 1,25-dihydroxyvitamin D3 receptors and activities in muscle. AB - Cytosols from cultured myoblast cells (G-8 and H9c2) prepared in high salt (0.3 M KCl) possesses receptor like proteins for 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) that sediment in the 3.2 S region of sucrose gradients. These receptors were characterized as having high affinity (Kd less than 0.1 nM) for 1,25-(OH)2D3 and are in low capacity (less than 80 fmol/mg of cytosol protein). Analog competition for receptor binding revealed that 1,25-(OH)2D3 was more potent than 24,25 (OH)2D3, or 25-(OH)2D3 for displacement of 1,25-(OH)2[3H]D3 from these 3.2 S region sedimenting receptors. Furthermore, the receptor proteins had affinity for DNA and eluted from Sephacryl S-200 as a macromolecule with Stokes radius (Rs) of 32 A. High salt cytosol from collagenase-dispersed skeletal muscle cells was also found to possess a 3.2 S 1,25-(OH)2D3 receptor-like protein. The 1,25-(OH)2D3 receptor concentration in both G-8 and H9c2 myoblast lines was found to down regulate by 50-70% when cells were stimulated to differentiate to myotubes by lowering fetal calf serum to 5% of the medium. Moreover, we demonstrated that 1,25-(OH)2D3 can inhibit DNA synthesis and cell proliferation of the G-8 myoblast cells in a dose-dependent manner. 1,25-(OH)2D3 was more potent at inhibiting cell proliferation in cells grown in 5% serum than in 20% serum. The data suggest that 1,25-(OH)2D3 can act directly on muscle myoblast via a 1,25-(OH)2D3 receptor that is similar to those found in intestine and bone. The data support the possibility that muscle is a target tissue for 1,25-(OH)2D3 and the hormone may act to initiate terminal differentiation of myoblast cells. PMID- 2991225 TI - Nucleotide sequence and gene organization of ColE1 DNA. AB - The primary structure of the plasmid ColE1 DNA has been determined. The plasmid DNA consists of 6646 base pairs (molecular mass of 4.43 MDa) and is 48.46% in GC content. The phi 80 trp insert of the composite plasmid of ColE1, pVH51, has also been determined. The determination of the nucleotide sequence of ColE1 DNA provides the basis for examining the relationships between the DNA sequence and the gene organization of the plasmid. The focus of this paper is to use this sequence data coupled with a review of the literature and our own work to examine the nine known functional regions of ColE1: imm (colicin E1 immunity), rep (replication function), inc (plasmid incompatibility and copy number control), bom (basis of mobility), rom (modulator of inhibition of primer formation by RNA I), mob (plasmid mobilization), cer (determinant for conversion of plasmid multimers to monomers), exc (plasmid entry exclusion), cea (structural gene for colicin E1), and kil (structural gene for the Kil protein). PMID- 2991226 TI - Oligonucleotides covalently linked to intercalating agents. Influence of positively charged substituents on binding to complementary sequences. AB - A pentamethylene chain was used to covalently link the 3'-phosphate of oligothymidylates to the 9-amino group of an acridine derivative. Positively charged substituents were further attached to the 3'-phosphate group to form 3' phosphotriesters. These molecules form specific complexes with poly(rA) which involve the formation of a number of A X T base pairs equal to that of thymines in the oligonucleotide. Absorption changes induced in the acridine absorption bands are similar to those expected upon intercalation of the acridine dye between A X T base pairs. The acridine covalently linked to the 3'-phosphate strongly stabilizes the complexes formed with poly(rA) as compared with the corresponding unsubstituted oligodeoxynucleotide. The presence of a positively charged substituent on the 3'-phosphate together with the acridine dye further enhances the interaction. The effect of salt concentration on complex stability depends on the number of negatively charged phosphate groups of the oligodeoxynucleotide and on the nature of the substituents borne by the 3' phosphate group. When the oligothymidylate is substituted by an acridine dye, the stability of the poly(rA) complexes increases when salt concentration increases. If an additional positively charged substituent is present on the 3'-phosphate group, stability decreases when salt concentration increases for the shortest oligonucleotide (trimer) and increases with longer oligonucleotides. Thermodynamic parameters have been calculated from the concentration dependence of melting temperatures. PMID- 2991227 TI - Electron paramagnetic resonance studies on the mechanism of activation and the catalytic cycle of the nickel-containing hydrogenase from Desulfovibrio gigas. AB - Desulfovibrio gigas hydrogenase (EC 1.12.2.1) is a complex enzyme containing one nickel, one 3Fe, and two [Fe4S4] clusters (Teixeira, M., Moura, I., Xavier, A. V., Der Vartanian, D. V., LeGall, J., Peck, H. D., Jr., Huynh, B. H., and Moura, J. J. G. (1983) Eur. J. Biochem. 130, 481-484). This hydrogenase belongs to a class of enzymes that are inactive "as isolated" (the so-called "oxygen-stable hydrogenases") and must go through an activation process in order to express full activity. The state of characterization of the active centers of the enzyme as isolated prompted us to do a detailed analysis of the redox patterns, activation profile, and catalytic redox cycle of the enzyme in the presence of either the natural substrate (H2) or chemical reductants. The effect of natural cofactors, as cytochrome C3, was also studied. Special focus was given to the intermediate redox species generated during the catalytic cycle of the enzyme and to the midpoint redox potentials associated. The available information is discussed in terms of a "working hypothesis" for the mechanism of the [NiFe] hydrogenases from sulfate reducing organisms in the context of activation process and catalytic cycle. PMID- 2991228 TI - Relationships between in vitro selenium supply, glutathione peroxidase activity, and phagocytic function in the HL-60 human myeloid cell line. AB - Utilizing the HL-60 human promyelocytic leukemia cell line cultured in defined medium, we examined the quantitative and temporal relationships between Se supply and the activity of the selenoenzyme glutathione peroxidase, as well as the effects of selenium deficiency on phagocytic function. Glutathione peroxidase activity depended on the medium Se concentration up to 2.6 X 10(-8) M (sodium selenate, 5 ng/ml), above which a plateau occurred. HL-60 cells grown in medium without Se supplementation became GSH peroxidase deficient, with activity 1-3% that of Se-replete cells. Replenishment of the medium with sodium selenate returned enzyme activity to 23% that of replete cells by 24 h and to 85% by 7 days, a process blocked by cycloheximide. Se-deficient HL-60 cells induced to granulocytic differentiation by dimethylformamide showed decreased hexose monophosphate shunt activity in response to phorbol myristate acetate and to an exogenous enzymatic H2O2-generating system. However, Se-deficient and -replete cells showed equal responses to methylene blue, which stimulates the shunt independently from the glutathione cycle. Se-deficient mature HL-60 cells stimulated with phorbol myristate acetate released 2.3-fold more H2O2 than Se replete cells and only slightly (not significantly) less O2. Se-deficient and replete differentiated HL-60 cells did not differ significantly in their capacities for cell motility or for ingestion of serum-opsonized bacteria. Differences between the findings of the present study and previous in vivo rat studies may reflect both the defined in vitro environment of the cell line and the inverse ratios of catalase and glutathione peroxidase activities in human and rat granulocytes. PMID- 2991229 TI - Regulation of pro-opiomelanocortin synthesis by dopamine and cAMP in the amphibian pituitary intermediate lobe. AB - The modulation of pro-opiomelanocortin (POMC) synthesis in Xenopus laevis pituitary intermediate lobe (IL) during background adaptation and the role of dopamine and cAMP in mediating this effect were examined. Neurointermediate lobes (NILs) were pulselabeled in vitro with [3H]arginine and analyzed for POMC synthesis by acid-urea gel electrophoresis. After black background adaptation of the animal (7 days), POMC synthesis increased 5-6-fold, while after white background adaptation (7 days), POMC synthesis decreased by 76%. Dopamine (50 microM) suppressed POMC synthesis in NILs in culture. In the absence of dopamine, POMC synthesis was stimulated. Several experiments were conducted to determine the category of dopamine receptor in the X. laevis IL. A D-2 dopamine receptor agonist inhibited immunoreactive alpha-MSH release from the NIL in a D-2 antagonist-reversible manner. A D-1 receptor agonist or antagonist did not alter the release of immunoreactive alpha-MSH from the NIL. Dopamine (10 microM) inhibited forskolin-stimulated cAMP accumulation. In addition, dopamine inhibition of POMC synthesis in cultured ILs was reversed by 8-Br-cAMP. These studies suggest that white background adaptation results in stimulation of the X. laevis D-2 receptor, which reduces cAMP production and POMC synthesis. Conversely, during black background adaptation the IL D-2 receptor is not stimulated, leading to increased cAMP production and POMC synthesis. PMID- 2991231 TI - Relationship between bacteriophage T4 and T6 DNA topoisomerases. T6 39-protein subunit is equivalent to the combined T4 39- and 60-protein subunits. AB - T6 DNA topoisomerase has been purified from bacteriophage T6 infected Escherichia coli. Unlike the T4 DNA topoisomerase which has three subunits, it consists of two subunits of molecular weights 75,000 and 51,000. They are the products of T6 genes 39 and 52, respectively. The purified T6 enzyme can stimulate in vitro T6 DNA replication. It has an ATP-dependent DNA relaxation activity similar to the T4 enzyme. Either ATP or dATP can be used in both reactions. Using a "Western blotting" and radioimmuno-detection methods, we show that T6 39 subunit contains protein sequences specified by both the T4 39 and 60 genes. The 52-proteins of both phages appear to be identical. The T4 and T6 topoisomerase genes represent a naturally occurring example of gene separation or fusion. PMID- 2991230 TI - Sequences of three closely related variants of a complex satellite DNA diverge at specific domains. AB - Major differences among the sequences of the repeat units of a very complex satellite DNA are located in domains which are sensitive to S1 nuclease under torsional stress, indicating that the domains assume unusual secondary or tertiary structures. Repeat units of the satellite, which accounts for 3% of the DNA of a land crab, have been inserted into pBR322 and the primary sequences of three cloned variants determined. The variants selected for sequencing include 1) RU (2089 base pairs (bp) ), representative of the average size of repeat units of cellular satellite; 2) TRU (1674 bp), truncated at an extra EcoRI site; and 3) EXT (2639 bp), extended by a 5-fold amplification of a 142-bp segment, one copy of which is present in RU and TRU (Bonnewell, V., Fowler, R.F., and Skinner, D.M. (1983) Science 221, 862-865). It appears that every copy of the satellite may be different and that the variants do not arise from cloning accidents. Extensive domains, as long as approximately 560 bp, are greater than 95% homologous among RU, TRU, and EXT; these conserved domains are composed of DNA whose base composition and sequences do not have remarkable features. By contrast, the sequences that comprise the divergent domains are unusually rich in 1) tracts of (dG X dC) 13-23 and arrangements of similar but not identical repetitive oligonucleotides or 2) alternating purines and pyrimidines (pu/py). PMID- 2991232 TI - Constitutively phosphorylated residues in the NS protein of vesicular stomatitis virus. AB - The NS protein of vesicular stomatitis virus is an auxiliary protein in the virus core (nucleocapsid) that plays a role in virus-specific RNA synthesis. NS exhibits a variety of phosphorylated forms, and the degree of phosphorylation correlates with the rate of RNA synthesis. However, chymotryptic peptide mapping has indicated that all forms of NS share a common cluster of phosphorylated residues. To locate these residues in the primary structure of the molecule, we performed a series of residue-specific chemical and enzymatic cleavages and separated radiophosphate-labeled peptides by gel electrophoresis. The data indicate that the constitutively phosphorylated sites in NS molecules reside in the amino-terminal region of the molecule, between residues 35 and 78. The previously reported resistance of the phosphoamino acids in this region to dephosphorylation by exogenous phosphatase suggests that this domain is embedded within the tertiary structure of the molecule or involved in quaternary interactions. In contrast, the amino acid residues that are phosphorylated secondarily, making NS more active in RNA synthesis, reside in more exposed regions of the molecule. PMID- 2991233 TI - Proteolytic activation of calmodulin-dependent cyclic nucleotide phosphodiesterase. AB - Purified calmodulin-stimulated cyclic nucleotide phosphodiesterase from brain, a homodimer of 59-kDa subunits, was activated by limited proteolysis with trypsin, alpha-chymotrypsin, Pronase, or papain and could not be further stimulated by addition of Ca2+ and calmodulin. Proteolysis increased Vmax and had little effect on the Km for cGMP. Treatment with alpha-chymotrypsin in the presence of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) produced, sequentially, 57- and 45-kDa peptides from the bovine and 55-, 53-, and 38-kDa peptides from the ovine enzyme. This protease-treated phosphodiesterase exhibited a Stokes radius of 3.9 nm and an S20,w value of 4.55; comparison with the hydrodynamic properties observed for native enzyme (4.3 nm, 5.95 S) strongly suggests a dimeric protein of Mr approximately 80,000-90,000. The proteolyzed species does not interact significantly with calmodulin immobilized on agarose, nor does it show complex formation with 2-dimethylaminonaphthalene-1-sulfonyl calmodulin even at micromolar concentrations of protein. Proteolysis, in the presence of calmodulin plus Ca2+, fully activated phosphodiesterase, producing the same intermediate peptides; however, final peptides from the bovine and ovine enzymes were 47 and 42 kDa, respectively, indicating a new, specific conformation of the enzyme. When EGTA was added to such incubations, these peptides were cleaved to those of the size seen when proteolysis was carried out entirely in the presence of EGTA. The initial rate of activation was increased by the presence of Ca2+ and calmodulin, suggesting that, in complex, phosphodiesterase exhibits a site with increased susceptibility to proteolysis. Since calmodulin can still interact with a fully activated form of the enzyme, it appears that retention of calmodulin binding can occur concomitantly with damage to that portion of the phosphodiesterase molecule responsible for suppression of its basal catalytic activity. PMID- 2991234 TI - Differential phosphorylation of multiple sites in protein 4.1 and protein 4.9 by phorbol ester-activated and cyclic AMP-dependent protein kinases. AB - The phosphorylation of the membrane skeleton components protein 4.1 and protein 4.9 in intact erythrocytes is shown to increase in the presence of either 1 microM 12-O-tetradecanoyl phorbol 13-acetate or 2 mM dibutyryl cAMP. The phosphorylation induced by these protein kinase activators is compared by two dimensional tryptic peptide mapping. In both proteins, the pattern of peptides phosphorylated in the presence of 12-O-tetradecanoyl phorbol 13-acetate differs from the pattern of peptides phosphorylated in the presence of dibutyryl cAMP. The relative locations of the phosphorylated sites on protein 4.1 have been determined using limited proteolysis by alpha-chymotrypsin. PMID- 2991235 TI - Photoreceptor GTP binding protein mediates fluoride activation of phosphodiesterase. AB - In this report, we show that fluoride activates dark-adapted rod outer segment phosphodiesterase, and that this activation is mediated, in analogy with adenylate cyclase, through a GTP binding protein. The GTP binding protein is released from dark-adapted rod outer segment membranes by exposure to fluoride and subsequent centrifugation. The 39-kilodalton subunit of the GTP binding protein, released from the membrane by this procedure, exhibits altered susceptibility to limited trypsin proteolysis, identical to that seen when hydrolysis-resistant GTP analogs are bound to that subunit. Repeated exposure of dark-adapted rod outer segment membranes to fluoride and subsequent centrifugation results in maximal activation of the membrane-bound phosphodiesterase. Thus, activation of phosphodiesterase by fluoride in the dark appears similar to fluoride activation of adenylate cyclase. PMID- 2991236 TI - Inositol 1,4,5-trisphosphate and the endoplasmic reticulum Ca2+ cycle of a rat insulinoma cell line. AB - Regulation of endoplasmic reticulum (ER) Ca2+ cycling by inositol 1,4,5 trisphosphate (IP3) was studied in saponin-permeabilized RINm5F insulinoma cells. Cells were incubated with mitochondrial inhibitors, and medium Ca2+ concentration established by nonmitochondrial pool(s) (presumably the ER) was monitored with a Ca2+ electrode. IP3 degradation accounted for the transience of the Ca2+ response induced by pulse additions of the molecule. To compensate for degradation, IP3 was infused into the medium. This resulted in elevation of [Ca2+] from about 0.2 microM to a new steady state between 0.3 and 1.0 microM, depending on both the rate of IP3 infusion and the ER Ca2+ content. The elevated steady state represented a bidirectional buffering of [Ca2+] by the ER, as slight displacements in [Ca2+], by small aliquots of Ca2+ or the Ca2+ chelator quin 2, resulted in net uptake or efflux of Ca2+ to restore the previous steady state. When IP3 infusion was stopped, [Ca2+] returned to its original low level. Ninety per cent of the Ca2+ accumulated by the ER was released by IP3 when the total Ca2+ content did not exceed 15 nmol/mg of cell protein. Above this high Ca2+ content, Ca2+ was accumulated in an IP3-insensitive, A23187-releasable pool. The maximal amount of Ca2+ that could be released from the ER by IP3 was 13 nmol/mg of cell protein. The data support the concept that in the physiological range of Ca2+ contents, almost all the ER is an IP3-sensitive Ca2+ store that is capable of finely regulating [Ca2+] through independent influx (Ca2+-ATPase) and efflux (IP3-modulated component) pathways of Ca2+ transport. IP3 may continuously modulate Ca2+ cycling across the ER and play an important role in determining the ER Ca2+ content and in regulating cytosolic Ca2+ under both stimulated and possibly basal conditions. PMID- 2991237 TI - The effects of alpha-adrenergic stimulation on the regulation of the pyruvate dehydrogenase complex in the perfused rat liver. AB - The regulation of the pyruvate dehydrogenase multienzyme complex was investigated during alpha-adrenergic stimulation with phenylephrine in the isolated perfused rat liver. The metabolic flux through the pyruvate dehydrogenase reaction was monitored by measuring the production of 14CO2 from infused [1-14C] pyruvate. In livers from fed animals perfused with a low concentration of pyruvate (0.05 mM), phenylephrine infusion significantly inhibited the rate of pyruvate decarboxylation without affecting the amount of pyruvate dehydrogenase in its active form. Also, phenylephrine caused no significant effect on tissue NADH/NAD+ and acetyl-CoA/CoASH ratios or on the kinetics of pyruvate decarboxylation in 14CO2 washout experiments. Phenylephrine inhibition of [1-14C]pyruvate decarboxylation was, however, closely associated with a decrease in the specific radioactivity of perfusate lactate, suggesting that the pyruvate decarboxylation response simply reflected dilution of the labeled pyruvate pool due to phenylephrine-stimulated glycogenolysis. This suggestion was confirmed in additional experiments which showed that the alpha-adrenergic-mediated inhibitory effect on pyruvate decarboxylation was reduced in livers perfused with a high concentration of pyruvate (1 mM) and was absent in livers from starved rats. Thus, alpha-adrenergic agonists do not exert short term regulatory effects on pyruvate dehydrogenase in the liver. Furthermore, the results suggest either that the rat liver pyruvate dehydrogenase complex is insensitive to changes in mitochondrial calcium or that changes in intramitochondrial calcium levels as a result of alpha-adrenergic stimulation are considerably less than suggested by others. PMID- 2991238 TI - Purification and characterization of a low molecular weight transforming growth factor from the urine of melanoma patients. AB - A low Mr human transforming growth factor (TGF) present in melanoma patients' urine has been purified approximately 200,000-fold to apparent homogeneity. Initial purification of an acid-soluble fraction of urine was achieved by Bio-Gel P-30 gel filtration chromatography in 1 M acetic acid. TGF activities were demonstrated in the Mr ranges of 30,000 and 6,000-10,000. These competed with epidermal growth factor (EGF) for binding to A431 membrane receptors and induced anchorage-independent growth of untransformed fibroblasts. The low Mr TGF activity obtained from P-30 chromatography was purified to apparent homogeneity by two sequential reverse-phase high performance liquid chromatography steps with a mu Bondapak C18 column first using a linear gradient of acetonitrile going from 0-60% in 120 min and then by rechromatography of the activity over the same column using a shallower gradient of acetonitrile going from 20-40% in 160 min. The isoelectric point of the melanoma patient-derived urinary TGF was determined to be 6.2, which is distinct from that for human EGF. Amino acid composition analysis of the purified urinary TGF (uTGF) revealed that it is composed of at least 42 amino acid residues with a minimum estimated Mr of 4,545. Compositional analysis further revealed distinct similarities and differences between the uTGF, human EGF and TGFs secreted by various transformed human and rodent cell lines. PMID- 2991239 TI - Kinetic identification of a two-state glucagon receptor system in isolated hepatocytes. Interconversion of homogeneous receptors. AB - A detailed kinetic study was performed to investigate the interaction of glucagon with receptors on freshly isolated hepatocytes. Competition binding assay results fit a mathematical expression for a single site noncooperative model of binding. Glucagon was shown to bind with first-order kinetics at six-hormone concentrations (0.02-0.50 nM) at 0 and 37 degrees C. The observed pseudo-first order rate constants are directly proportional to the hormone concentration at 0 degree C, but display a downward deviation from linearity at 37 degrees C. Dissociation of glucagon exhibited biexponential character at 37 degrees C which was not seen at 0 degree C. The biphasic dissociation at 37 degrees C was resolved into rapid (t1/2 = 1.9 min) and slow (t1/2 = 27.7 min) components. The distribution of the total bound hormone between the rapidly and slowly dissociating complexes was not dependent upon the extent of receptor occupancy. The absolute quantity of rapidly dissociating hormone-receptor complexes was constant at all times examined; however, the fraction of slowly dissociating hormone-receptor complexes was found to increase with increasing incubation time. The results indicate that a homogeneous population of hepatic receptors undergoes a time-dependent, temperature-dependent conversion from one state to another in a two-stage sequential manner. PMID- 2991240 TI - Replication of pBR322 DNA in vitro with purified proteins. Requirement for topoisomerase I in the maintenance of template specificity. AB - The replication of plasmid pBR322 DNA has been reconstituted with purified proteins from Escherichia coli. Initiation of the leading-strand requires RNA polymerase holoenzyme, DNA polymerase I, RNase H, and DNA gyrase. Initiation of the lagging-strand requires the primosomal proteins (the dnaB, dnaC, and dnaG proteins, replication factor Y (protein n') and proteins i, n, and n") and the single-stranded DNA binding protein. DNA polymerase III holoenzyme is required for extensive elongation of the nascent DNA chains. The products of this replication reaction are primarily nonsegregated daughter molecules. However, the addition of small amounts of soluble extract from E. coli results in the completion and segregation of these molecules to give mature form I DNA, suggesting that additional factors are required for this process. Topoisomerase I is necessary to make the replication system specific for pBR322 DNA as a template, indicating that the linking number of the DNA, determined by an equilibrium between the opposing activities of topoisomerase I and DNA gyrase, plays a crucial role in determining the reactivity of the DNA molecule toward initiating DNA replication. The function of the proteins involved in the replication of this closed-circular, double-stranded, superhelical DNA is discussed. PMID- 2991241 TI - Characterization and derivation of the gene coding for mitochondrial carbamyl phosphate synthetase I of rat. AB - The nucleotide sequence of rat carbamyl phosphate synthetase I mRNA has been determined from the complementary DNA. The mRNA comprises minimally 5,645 nucleotides and codes for a polypeptide of 164,564 Da corresponding to the precursor form of the rat liver enzyme. The primary sequence of mature rat carbamyl phosphate synthetase I indicates that the precursor is cleaved at one of two leucines at residues 38 or 39. The derived amino acid sequence of carbamyl phosphate synthetase I is homologous to the sequences of carbamyl phosphate synthetase of Escherichia coli and yeast. The sequence homology extends along the entire length of the rat polypeptide and encompasses the entire sequences of both the small and large subunits of the E. coli and yeast enzymes. The protein sequence data provide strong evidence that the carbamyl phosphate synthetase I gene of rat, the carAB gene of E. coli, and the CPA1 and CPA2 genes of yeast were derived from common ancestral genes. Part of the rat carbamyl phosphate synthetase I gene has been characterized with two nonoverlapping phage clones spanning 28.7 kilobases of rat chromosomal DNA. This region contains 13 exons ranging in size from 68 to 195 base pairs and encodes the 453 carboxyl-terminal amino acids of the rat protein. Southern hybridization analysis of rat genomic DNA indicates the carbamyl phosphate synthetase I gene to be present in single copy. PMID- 2991242 TI - The association of the interspersed repetitive KpnI sequences with the nuclear matrix. AB - The KpnI sequences constitute the dominant, long, interspersed repetitive DNA families in primate genomes. These families contain related, but nonidentical sequence subsets, some of which border functional gene domains and are transcribed into RNA. To test whether these sequences perform an organizational function in the nucleus, their association with the nuclear matrix has been examined in African green monkey cells. DNase I treatment depleted the residual matrix of most of the KpnI 1.2- and 1.5-kilobase pair family sequences although significant amounts of each family remained in the loop attachment DNA fragments. Hybridization analysis of the KpnI and RsaI cleavage patterns of matrix loop attachment DNA indicate that some sequence subsets of these KpnI families are relatively less depleted than others. The nuclear matrix association of subpopulations of KpnI 1.2- and 1.5-kilobase pair families was also shown by metrizamide gradient centrifugation of nuclear matrix complexes cleaved by KpnI endonuclease. The gradients demonstrate that some KpnI segments are differentially associated with nuclear matrix proteins. Moreover, the procedures permit the preparative isolation and purification of the DNA-protein complexes containing these KpnI 1.2- and 1.5-kilobase pair sequence families. Speculations on the relationship between the matrix association of these KpnI family sequences and their possible roles in gene organization and expression are presented and discussed. PMID- 2991243 TI - Crystallization and preliminary x-ray investigation of the regulatory subunit of cAMP-dependent protein kinase. AB - Crystals of type I cAMP-dependent protein kinase regulatory subunit have been grown from solutions of ammonium sulfate. The crystals are square bipyramids, space group P4(1)2(1)2 (P4(3)2(1)2), with a = b = 106.9 +/- 0.6 A and c = 212.4 +/- 1.0 A. There are two dimers of the regulatory subunit/crystallographic asymmetric unit. The crystals are stable for 3-4 days in the x-ray beam and diffract to at least 3.5-A resolution. PMID- 2991244 TI - Mechanism of phorbol diester-induced regulation of surface transferrin receptor involves the action of activated protein kinase C and an intact cytoskeleton. AB - Phorbol diesters are tumor-promoting agents that cause differentiation of HL60 human leukemic cells and concomitantly regulate surface transferrin receptors. Regulation of transferrin receptors by phorbol diesters involves receptor internalization in association with increased receptor phosphorylation (hyperphosphorylation). The intracellular mechanism of action of phorbol diester involves binding to and activation of the Ca2+-phospholipid-dependent protein kinase (protein kinase C). Present studies comparing results obtained with whole cells and those from a cell-free system reconstituted from purified protein kinase C and transferrin receptor components have revealed that the transferrin receptor is phosphorylated by protein kinase C activated by phorbol esters. Following tryptic digestion and two-dimensional separation of phosphopeptides of phosphorylated transferrin receptors, two major and several minor phosphoserine containing fragments are resolved. These fragments are identical whether transferrin receptor is phosphorylated in whole cells incubated with phorbol diesters or following phosphorylation of affinity immobilized transferrin receptor in the in vitro reconstitution system. Phosphoamino acid analysis of these fragments indicates that serine is the only amino acid phosphorylated in whole cells or in the cell-free system. In addition, colchicine is shown to inhibit in a dose-dependent manner phorbol diester-induced internalization but not hyperphosphorylation of the surface transferrin receptor in whole cells. This inhibition is specific for colchicine since inactive beta- and gamma Lumicolchicine have no such effect, while taxol reverses the inhibition. These results indicate that the phorbol diester-mediated process of down-regulation of the surface transferrin receptor is associated with phosphorylation of the receptor by activated protein kinase C and requires an intact cytoskeleton to affect receptor internalization. PMID- 2991245 TI - The succinate dehydrogenase of Escherichia coli. Immunochemical resolution and biophysical characterization of a 4-subunit enzyme complex. AB - Using EPR spectroscopy to monitor the integrity of the enzyme, conditions have been established which allow specific immunoprecipitation of the succinate dehydrogenase complex of Escherichia coli. The enzyme complex precipitated from Lubrol PX-solubilized membranes by monospecific antiserum in the presence of a cocktail of protease inhibitors contains four polypeptides of apparent MrS 71,000, 26,000, 17,000, and 15,000. The 71-kDa flavopeptide is readily susceptible to proteolysis, and the enzyme complex shows unusual facile dissociation. Spectroscopic measurements indicate the presence of a [2Fe-2S] cluster (Center 1), a [3Fe-xS] cluster (Center 3), and a b-type cytochrome. In addition, a change in relaxation of Center 1 at low potentials is indicative of Center 2. Midpoint redox potentials of Centers 1-3 for both the membrane-bound and detergent-solubilized enzyme were estimated to be +10 mV, -175 mV, and +65 mV, respectively. PMID- 2991246 TI - Direct demonstration of rapid insulin-like growth factor II Receptor internalization and recycling in rat adipocytes. Insulin stimulates 125I-insulin like growth factor II degradation by modulating the IGF-II receptor recycling process. AB - The photoactive insulin-like growth factor (IGF)-II analogue 4-azidobenzoyl-125I IGF-II was synthesized and used to label specifically and covalently the Mr = 250,000 Type II IGF receptor. When rat adipocytes are irradiated after a 10-min incubation with 4-azidobenzoyl-125I-IGF-II at 10 degrees C and immediately homogenized, most of the labeled IGF-II receptors are associated with the plasma membrane fraction, indicating that receptors accessible to the labeling reagent at low temperature are on the cell surface. However, when the photolabeled cells are incubated at 37 degrees C for various times before homogenization, labeled IGF-II receptors are rapidly internalized with a half-time of 3.5 min as evidenced by a loss from the plasma membrane fraction and a concomitant appearance in the low density microsome fraction. The low density microsomes were previously shown to contain intracellular membranes (Oka, Y., and Czech, M.P. (1984) J. Biol. Chem. 259, 8125-8133). The steady state level of cell surface IGF II receptors in the presence or absence of IGF-II, measured by the binding of anti-IGF-II receptor antibody to cells, remains constant under these conditions, demonstrating that IGF-II receptors rapidly recycle back to the cell surface at the same rate as receptor internalization. Using the above methodology, it is shown that acute insulin action: 1) increases the steady state number of cell surface IGF-II receptors; 2) increases the number of ligand-bound IGF-II receptors that are internalized per unit of time, as evidenced by a large increase in the photolabeling of intracellular membrane IGF-II receptors when cells are incubated at 37 degrees C with insulin and 4-azidobenzoyl-125I-IGF-II prior to photoactivation; and 3) increases the rate of cellular 125I-IGF-II degradation by a process that is blocked by anti-IGF-II receptor antibody. The results indicate that the action of insulin to elevate the steady state number of cell surface IGF-II receptors leads to an increased internalization flux of IGF II-bound receptors, mediating increased IGF-II uptake and degradation. PMID- 2991247 TI - Studies on K+ permeability of rat gastric microsomes. AB - A population of gastric membrane vesicles of high K+ permeability and of lower density than endoplasmic tubulovesicles containing (H+-K+)-ATPase was detected in gastric mucosal microsomes from the rat fasted overnight. The K+-transport activity as measured with 86RbCl uptake had a Km for Rb+ of 0.58 +/- 0.11 mM and a Vmax of 13.7 +/- 1.9 nmol/min X mg of protein. The 86Rb uptake was reduced by 40% upon substituting Cl- with SO2-4 and inhibited noncompetitively by ATP and vanadate with a Ki of 3 and 30 microM, respectively; vanadate also inhibited rat gastric (H+-K+)-ATPase but with a Ki of 0.03 microM. Carbachol or histamine stimulation decreased the population of the K+-permeable light membrane vesicles, at the same time increased K+-transport activity in the heavy, presumably apical membranes of gastric parietal cells, and enabled the heavy microsomes to accumulate H+ ions in the presence of ATP and KCl without valinomycin. The secretagogue-induced shift of K+ permeability was blocked by cimetidine, a H2 receptor antagonist. Four characteristics of the K+ permeability as measured with 86RbCl were common in the resting light and the carbachol-stimulated heavy microsomes; (a) Km for +Rb, (b) anion sensitivity (Cl- greater than SO2-4), (c) potency of various divalent cations (Hg2+, Cu2+, Cd2+, and Zn2+) to inhibit Rb+ uptake, and (d) inhibitory effect of ATP, although the nucleotide sensitivity was latent in the stimulated heavy microsomes. The Vmax for 86RbCl uptake was about 10 times greater in the resting light than the stimulated heavy microsomes. These observations led us to propose that secretagogue stimulation induces the insertion of not only the tubulovesicles containing (H+-K+)-ATPase, but also the light membrane vesicles containing KCl transporter into the heavy apical membranes of gastric parietal cells. PMID- 2991248 TI - Cloning and characterization of the yeast nuclear gene for subunit 5 of cytochrome oxidase. AB - The nuclear gene COX5 coding for subunit 5 of cytochrome oxidase has been cloned by transformation of the cox5-1 mutant aE4-238/AL1 with a library of yeast genomic DNA. The recombinant plasmid pG46/ST2 bearing a nuclear DNA insert of 1.17 kilobase pairs restores the ability of cox5 mutants to respire and to synthesize a wild type subunit 5. The COX5 gene has been sequenced and determined to code for a 153-amino acid long protein with a molecular weight of 17,121. The amino-terminal 20 residues comprise the signal peptide. The sequence starting from residue 21 matches the partial sequence reported for the mature subunit 5. The sequence of the subunit 5 gene indicates that the mature protein has a molecular weight of 14,858 which agrees with previous size estimates based on electrophoretic migration. The primary sequence and polarity profile of yeast subunit 5 establishes that it is homologous to subunit 4 of bovine cytochrome oxidase. PMID- 2991249 TI - Site-specific mutagenesis of ribulose-1,5-bisphosphate carboxylase/oxygenase. Evidence that carbamate formation at Lys 191 is required for catalytic activity. AB - Site-specific mutagenesis of a cloned gene for ribulose-1,5-bisphosphate carboxylase/oxygenase from Rhodospirillum rubrum was used to examine the functional significance of carbamate activation. Lysine 191, the residue involved in carbamate formation, was replaced with a glutamate in order to mimic the anionic nature of the carbamate. The resulting enzyme was capable of binding the six-carbon transition state analog carboxyarabinitol bisphosphate, but completely lacked catalytic activity. In contrast to the wild-type enzyme, carboxyarabinitol bisphosphate binding was not stabilized by divalent metal and CO2. These observations are consistent with a proposed role for the carbamate in binding the metal required for catalysis. PMID- 2991250 TI - Guanosine 5'-O-thiotriphosphate stimulates phospholipase C activity in plasma membranes of rat hepatocytes. AB - The guanine nucleotide analogue, guanosine 5'-O-thiotriphosphate (GTP gamma S) stimulated plasma membrane-associated phospholipase C. Phosphoinositides were the substrates for the reaction. Significant losses of phosphatidylinositol bisphosphate and phosphatidylinositol phosphate occurred at lower doses of GTP gamma S than did significant loss of phosphatidylinositol. Loss of 32P-labeled phosphatidylinositol bisphosphate was equal when plasma membranes were treated with either 100 microM GTP or 100 microM GTP gamma S, but accumulation of inositol trisphosphate was more apparent when the nonhydrolyzable analogue was used. The action of GTP gamma S alone was not dependent on Ca2+ although loss of 32P-labeled phosphoinositides was stimulated by Ca2+ alone or with GTP gamma S. The results are consistent with a role for guanine nucleotide binding proteins in the activation of membrane-bound phosphoinositide-specific phospholipase C. PMID- 2991251 TI - Stoichiometry determination for carbon monoxide binding to Rhodospirillum molischianum cytochrome c'. AB - The stoichiometry of CO ligation to the dimer heme protein Rhodospirillum molischianum cytochrome c' is determined. We have recently measured the enthalpy change of CO ligation to this molecule by the van't Hoff method and found the value of -10.7 +/- 1.2 kcal/mol CO (aqueous) (Doyle, M. L., Weber, P. C., and Gill, S. J. (1985) Biochemistry 24, 1987-1991). In the present paper the enthalpy change of CO ligation, measured directly by titration calorimetry, is found to be -9.5 +/- 0.2 kcal/mol heme. Since the van't Hoff method gives the heat value in units/mole of CO and the calorimetric method gives the heat value in units/mole of heme, the stoichiometry of the reaction is given by the ratio of the two values and found to be 0.9 +/- 0.1, or within experimental error, one CO molecule bound per heme. PMID- 2991252 TI - Nucleotide sequence of mouse prolactin and growth hormone mRNAs and expression of these mRNAs during pregnancy. AB - The mRNAs for mouse prolactin and growth hormone have been isolated from anterior pituitary glands and cloned as cDNAs. The nucleotide sequences of these mRNAs have been determined, and these sequences, along with the predicted amino acid sequences, are compared to those of other mammalian prolactin and growth hormone mRNAs. Levels of prolactin and growth hormone mRNAs during pregnancy have been monitored by hybridization to the cloned cDNA probes. We find the levels of these mRNAs to remain nearly constant during mid-to-late gestation. PMID- 2991253 TI - Intron structure of the human antithrombin III gene differs from that of other members of the serine protease inhibitor superfamily. AB - Antithrombin III (ATIII) plays an integral role in the coagulation system by inhibiting thrombin and several other activated clotting factors. Inherited deficiency of ATIII is quite common and can result in life-threatening thrombotic complications. In order to understand the basis of ATIII deficiency, we have isolated and characterized the normal human ATIII gene from a recombinant Charon 4A bacteriophage genomic library. The ATIII gene contains six exons and five introns distributed over approximately 19 kilobases of DNA. The positions of introns in the ATIII gene were compared with other members of the serine protease inhibitor family which share 17-31% amino acid homology. When aligned to achieve maximal protein homology, only one of the ATIII introns corresponded to the four introns of rat angiotensinogen or human alpha 1-antitrypsin. Similarly, only one ATIII intron was homologous to the seven introns of chicken ovalbumin. We present two testable models to explain the discrepancy in intron positions among members of the serine protease inhibitor superfamily of genes. PMID- 2991254 TI - Biphasic effects of type beta transforming growth factor on epidermal growth factor receptors in NRK fibroblasts. Functional consequences for epidermal growth factor-stimulated mitosis. AB - Exposure of confluent NRK cells to transforming growth factor-beta (TGF-beta) results in distinct alterations in subpopulations of plasma membrane epidermal growth factor (EGF) receptors. The low affinity sites increase in number, whereas the high affinity sites undergo a transient decrease in affinity followed by a prolonged increase in number. Cycloheximide inhibits both of these effects. Functional assays measuring EGF-stimulated thymidine incorporation in the presence of TGF-beta show that the resulting long-term stimulation of EGF receptor binding is associated with an increased sensitivity to EGF. Similarly, the initial, transient decrease in EGF binding is associated with a temporary inhibition of EGF-stimulated thymidine incorporation. The results describe a bifunctional effect of TGF-beta at the biochemical level consistent with the action of this peptide on NRK cell growth. PMID- 2991256 TI - Characterization of the oxidized states of bromoperoxidase. AB - Bromoperoxidase Compound I has been formed in reactions between bromoperoxidase and organic peroxide substrates. The absorbance spectrum of bromoperoxidase Compound I closely resembles the Compound I spectra of other peroxidases. The pH dependence of the second order rate constant for the formation of Compound I with hydrogen peroxide demonstrates the presence of an ionizable group at the enzyme active site having a pKa of 5.3. Protonation of this acidic group inhibits the rate of Compound I formation. This pKa value is higher than that determined for other peroxidases but the overall pH rate profiles for Compound I formation are similar. The one-electron reduction of bromoperoxidase Compound I yields Compound II and a second reduction yields native enzyme. Bromoperoxidase Compound II readily forms Compound III in the presence of an excess of hydrogen peroxide. Compound III passes through an as yet uncharacterized intermediate (III) in its decay to native enzyme. Compound III is produced and accumulates in enzymatic bromination reactions to become the predominate steady state form of the enzyme. Since Compound III is inactive as catalyst for enzymatic bromination, its accumulation leads to an idling reaction pathway which displays an unusual kinetic pattern for the bromination of monochlorodimedone. PMID- 2991255 TI - Nucleotide specificity of cardiac sarcoplasmic reticulum. GTP-induced calcium accumulation and GTPase activity. AB - We previously demonstrated that the hydrolysis of GTP by canine cardiac sarcoplasmic reticulum is not sensitive to calcium and does not support the translocation of calcium and oxalate into the vesicular space. In response to GTP, however, calcium is accumulated into a compartment which is sensitive to pH and ionophore. In the present paper, we further explored the relationship between GTP hydrolysis and GTP-induced calcium accumulation. Both ATP- and GTP-induced calcium accumulation were prevented by the sulfhydryl reagent, N-ethylmaleimide (NEM; I50 = 0.2 mM). In contrast, the sensitivity of NTP hydrolysis to NEM differed markedly; GTPase activity was not affected by NEM, whereas ATPase activity was markedly inhibited. Conversely, although the GTPase was noncompetitively inhibited by the ATP analogue, adenylyl imidodiphosphate (Ki = 8 microM), and was competitively inhibited by the GTP analogue, guanylyl imidodiphosphate (Ki = 60 microM), GTP-induced calcium accumulation was not affected by the NTP analogues at any concentration. Therefore, the GTP-dependent accumulation of calcium into the pH- and ionophore-sensitive compartment of cardiac SR may not require GTP hydrolysis but may be dependent on GTP binding. The previously reported noncompetitive inhibition of the GTPase by ATP was also observed when the calcium-dependent hydrolysis of ATP was prevented by NEM (Ki = 1.2 microM). Along with the noncompetitive inhibition of the GTPase by adenylyl imidodiphosphate, the inhibition of the GTP by ATP in the presence of NEM suggests that ATP binding may be involved in the observed inhibition. The Ki for the noncompetitive inhibition of GTPase activity is compatible with ATP binding to the high affinity catalytic site of the ATPase. Thus, although GTP-induced calcium accumulation differs somewhat from ATP-dependent calcium translocation, the similarities between the two processes (i.e. similar time courses and sensitivity to pH, ionophore, and sulfhydryl modification) suggest that they may be related in some manner. PMID- 2991257 TI - Effects of fatty acids on Na+-Ca2+ exchange and Ca2+ permeability of cardiac sarcolemmal vesicles. AB - We have previously reported that anionic phospholipids (Philipson, K.D., and Nishimoto, A.Y. (1984) J. Biol. Chem. 259, 16-19) and other anionic amphiphiles (Philipson, K.D. (1984) J. Biol. Chem. 259, 13999-14002) stimulate Na+-Ca2+ exchange in cardiac sarcolemmal vesicles. To further these studies, we have now investigated the effects of a variety of fatty acids on both Na+-Ca2+ exchange and passive Ca2+ permeability. Na+-Ca2+ exchange was stimulated by fatty acids by up to 150%. Unsaturated fatty acids were more potent than saturated fatty acids, and the stimulation was primarily due to a decrease in the apparent KM (Ca2+). There was a positive correlation between the ability of a fatty acid to stimulate Na+-Ca2+ exchange and to increase passive Ca2+ permeability. The methyl esters of fatty acids had no effects on either exchange or permeability indicating the importance of anionic charge. We conclude that the combination of local lipid disorder and anionic charge regulate Na+-Ca2+ exchange. Perturbations of the bilayer hydrophobic region and increased negative surface charge are both required for fatty acids to increase passive Ca2+ flux. Na+-Ca2+ exchange is stimulated when the ratio of membrane free fatty acid to phospholipid is about 5%. This level of fatty acid is achieved during 1 h of myocardial ischemia (Chien, K. R., Han, A., Sen, A., Buja, L. M., and Willerson, J. T. (1984) Circ. Res. 54, 313-322), indicating that ischemia could induce altered sarcolemmal Ca2+ transport due to fatty acid accumulation. PMID- 2991258 TI - Identification of linoleic and oleic acids as endogenous Na+,K+-ATPase inhibitors from acute volume-expanded hog plasma. AB - Na+,K+-ATPase inhibitors have been found to exist in acutely saline-infused hog plasma, which also inhibit the specific binding of ouabain to Na+,K+-ATPase and the binding of digoxin to specific anti-digoxin antibody. Two of these inhibitors were purified by a combination of Amberlite XAD-2 adsorption chromatography and 3 steps of high-performance liquid chromatography. Reverse phase, high-performance liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectrometry identified these substances as linoleic (18:2) and oleic acids (18:1). A significant increase in the ouabain-displacing activity was observed in hog plasma during saline infusion. The maximal level reached was approximately 10 times higher than that of the preinfusion plasma sample. The two unsaturated fatty acids contributed to approximately 52% of the total ouabain-displacing activity after 120 min of saline infusion. The increased fatty acid levels in volume-expanded plasma are sufficient for an extensive inhibition of Na+,K+ ATPase activity. These results strongly suggest that free unsaturated fatty acids in plasma regulate extracellular fluid volume in a pathological volume-expanded condition through modulation of Na+,K+-ATPase activity. PMID- 2991259 TI - Biochemical properties of the Na+/H+ exchange system in rat brain synaptosomes. Interdependence of internal and external pH control of the exchange activity. AB - Properties of the Na+/H+ exchange system in synaptosomes have been studied primarily by using acridine orange fluorescence to follow H+ efflux. Results obtained from 22Na+ uptake experiments and [3H]ethylpropylamiloride binding experiments are also presented for comparison. The basal properties of the Na+/H+ antiport in synaptosomes are similar to those found in other systems; (i) the stoichiometry of Na+/H+ exchange is 1:1; (ii) Li+ can be successfully substituted for Na+; its affinity for the exchanger (KLi+ = 3 mM) is higher than that of Na+ (KNa+ = 12 mM), but the maximal rate of H+ efflux in the presence of Li+ is about 3 times lower than the maximal rate of H+ efflux in the presence of Na+; and (iii) the Na+/H+ antiport is inhibited by amiloride derivatives with the rank order:ethylisopropylamiloride greater than ethylpropylamiloride greater than amiloride greater than benzamil. The most important finding of this paper is that the external pH dependence of the synaptosomal Na+/H+ antiport is controlled by the value of internal pH and vice versa. For example apparent pHo values for half maximum activation of the Na+/H+ exchanger are pHo = 7.12 when pHi = 6.4 and pHo = 7.95 when pHi = 7.3. Therefore, a 0.9 pH unit increase in internal pH produces a shift of at least a 0.83 pH unit in the external pH dependence. In addition, changing pHo from 7.75 to 8.50 also shifts the half-maximum pHi value for activation of the Na+/H+ antiport from 6.67 to 7.54. PMID- 2991260 TI - Sodium bicarbonate in seminal plasma stimulates the motility of mammalian spermatozoa through direct activation of adenylate cyclase. AB - Recently, a low molecular weight factor, which specifically stimulates sperm adenylate cyclase, was found in porcine seminal plasma (Okamura, N., and Sugita, Y. (1983) J. Biol. Chem. 258, 13056-13062). The purified factor was analyzed by 1H NMR, 13C NMR, infrared spectroscopy, and elementary analysis and identified as sodium bicarbonate. The effects of sodium bicarbonate both on adenylate cyclase activity in porcine spermatozoa and on sperm motility have been studied. Sperm adenylate cyclase was found to be specifically activated by bicarbonate over the physiological concentration range. In contrast, the adenylate cyclase activity in other tissues was not affected. The same concentration range of bicarbonate which resulted in activation of adenylate cyclase also stimulated sperm motility. The motility and enzyme activity of spermatozoa in all species so far tested (human, bovine, rat, mouse, and dog) were found to be similarly sensitive to bicarbonate concentration. These results show that the bicarbonate-sensitive adenylate cyclase system regulates sperm motility and suggest that this system is common to all mammals. PMID- 2991261 TI - Isolation of a specific mu-opiate receptor peptide, morphiceptin, from an enzymatic digest of milk proteins. AB - Specific radioimmunoassays have been developed for the measurement of naturally occurring morphiceptin and beta-casomorphin. These peptides and related exorphins were isolated from an enzymatic digest of caseins by chromatographic techniques including gel filtration, hydrophobic column and multiple-step high pressure liquid chromatography. Three exorphins were purified and characterized in their radioimmunological, biological, and chemical properties. They were identified as morphiceptin, beta-casomorphin, and 8-prolyl-beta-casomorphin. Since morphiceptin is a highly specific mu-agonist and can be derived from a milk protein, it is possible that morphiceptin is an exogenous opioid ligand specific for mu receptors in the brain and gastrointestinal tract. PMID- 2991262 TI - Accessibility of lysyl residues of Escherichia coli B/r porin (OmpF) to covalent labeling reagents of different sizes. An approach for a three-dimensional structure of a channel-forming protein. AB - The three-dimensional structure of Escherichia coli B/r porin (OmpF) was studied by chemical modification using activated sugars of different size. Galactose and galactosides of different penetration properties through the porin channel were oxidized by galactose oxidase, and the 6-aldehydes formed were linked to amino groups in porin by reduction with NaBH3CN. Tryptic fragments of modified and unmodified porin were separated by reversed-phase high pressure liquid chromatography and identified by amino acid and amino-terminal analysis from the known primary structure of OmpF. Modification of purified native porin trimers in beta-octylglucoside revealed three classes of amino groups: (i) those not modified by any sugars; (ii) those modified only by small sugars that diffuse rapidly through the pore, such as galactose or melibiose; and (iii) those modified by either small or large sugars, the latter including pore-impermeant sugars such as stachyose. The results suggest that the three classes of amino groups correspond, respectively, to groups buried in the trimeric molecule, those in the interior of the pore and those exposed on the surface of porin. In addition modification experiments performed on whole cells suggested that all the reactive groups modified by the pore-impermeant sugars (class iii) are located on the surface of porin exposed on the outside of the outer membrane. PMID- 2991263 TI - Characterization of insulin-like growth factor I-stimulated tyrosine kinase activity associated with the beta-subunit of type I insulin-like growth factor receptors of rat liver cells. AB - We previously reported that insulin-like growth factor I (IGF-I) stimulates the phosphorylation of a Mr 98,000 protein thought to be the beta-subunit of the type I IGF receptor of BRL-3A2 rat liver cells, as well as phosphorylation of the exogenous tyrosine-containing substrate poly(Glu,Tyr), 4:1. The present study provides additional evidence that the type I IGF receptor possesses intrinsic tyrosine kinase activity and characterizes the properties of this receptor kinase. IGF-I stimulates receptor phosphorylation and phosphorylation of poly(Glu,Tyr), 4:1, by lectin-purified receptor preparations with the same concentration dependence; half-maximal stimulation was observed with approximately 3 nM IGF-I and approximately 3-fold higher concentrations of insulin. Although IGF-I-dependent receptor phosphorylation was observed within 2 min and was maximal after 10 min, phosphorylation of exogenous substrate did not begin to increase until 8 min after addition of [gamma-32P] ATP and poly(Glu,Tyr), 4:1. When IGF-I-receptor complexes were preincubated with unlabeled ATP for 10 min before addition of substrate, however, IGF-I-dependent 32P incorporation was observed within 2 min after addition of poly(Glu,Tyr), 4:1, and increased linearly for 20 min. We propose that this activation of type I IGF receptor tyrosine kinase activity results from autophosphorylation of the receptor kinase. Kinase activation is an intramolecular reaction, is specific for ATP, occurs within 3 min after addition of unlabeled ATP, and requires the presence of IGF-I before or concomitant with activation by ATP. IGF-I acts rapidly, stimulating substrate phosphorylation within 3 min. The properties of the type I-IGF receptor kinase closely resemble those of the insulin receptor kinase, suggesting that the homologies between the two receptors extend to their kinase domains. PMID- 2991264 TI - Regulation of epidermal growth factor receptor by estrogen. AB - Administration of 17 beta-estradiol (E2) to immature female rats produces a 3 fold increase in 125I-epidermal growth factor (EGF) binding to uterine membranes with no change in the affinity of membrane receptors for EGF. E2 treatment also increases the EGF receptor visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after affinity labeling of uterine membranes and the EGF stimulated receptor autophosphorylation activity. In addition, E2 administration stimulates EGF-dependent tyrosine kinase activity in an assay system using exogenous angiotensin II as substrate. Following hormone treatment, EGF receptor levels increase between 6 and 12 h, remain elevated at 18 h, and decline between 24 and 36 h. This stimulation of EGF receptor levels by E2 is specific, since the non-estrogenic hormones progesterone, dexamethasone, and dihydrotestosterone fail to elevate receptor levels. E2-stimulated increases in EGF receptor levels are also blocked by cycloheximide and actinomycin D, suggesting that the observed effect represents de novo synthesis of the EGF receptor and may be mediated by a transcriptional mechanism. These results demonstrate that estrogen can regulate acutely the levels of EGF receptor in vivo and raise the possibility that events coupled to this receptor may play a role in estrogen-stimulated growth. PMID- 2991266 TI - Auxin-binding protein from coleoptile membranes of corn (Zea mays L.). I. Purification by immunological methods and characterization. AB - The purification of a putative auxin receptor is one possibility to elucidate the first event in the mechanism of auxin action. By affinity chromatography of membrane proteins on 2-OH-3,5-diiodobenzoic acid-Sepharose and gel filtration on Ultrogel a fraction enriched in auxin-binding protein (ABP) was obtained and used for rabbit immunization. From the immunoglobulin G (IgG) fraction of the antisera IgGs against proteins not binding auxin (nonABP) could be obtained which were used to eliminate the nonABP from the eluates of the 2-OH-3,5-diiodobenzoic acid Sepharose. The remainder fraction was further purified and concentrated on IgG Sepharose which retained the ABP that could be eluted without loss of binding activity. A 600-fold purification with a yield of 42% was achieved. The ABP could be identified as the site I "receptor" described by Dohrmann et al. (Dohrmann, U., Hertel, R., and Kowalik, H. (1978) Planta (Berl.) 140, 97-106). It is shown that the competitors tested reduce [14C]1-naphthylacetic acid-(NAA) binding in the following order of effectiveness: NAA greater than 2-naphthylacetic acid greater than 1-phenylacetic acid greater than 2,3,5-triiodobenzoic acid greater than 3-indolylacetic acid greater than 2,4-dichlorophenoxyacetic acid. The ABP has a sharp binding optimum at pH 5.5, and the KD was calculated to be 5.7 X 10( 8) M to [14C]NAA. The binding activity of the ABP linearly decreased with increasing temperature but could partially be restored upon chilling in the presence of auxin. The ABP seems to be a 40-kDa dimer in its native form without disulfide bonds between its monomers. PMID- 2991265 TI - A rat brain isozyme of angiotensin-converting enzyme. Unique specificity for amidated peptide substrates. AB - We have purified angiotensin-converting enzyme (ACE, EC 3.4.15.1) from rat brain corpus striatum and rat lung. The brain enzyme has Mr 165,000 by sodium dodecyl sulfate gel electrophoresis, whereas the lung enzyme is 175,000. This difference is not an artifact of preparation since mixture of the two tissues prior to purification results in isolation of two proteins with Mr 165,000 and 175,000. Separation of tryptic fragments of 125I-labeled lung and brain ACE by reverse phase chromatography yields distinct but similar patterns. No differences between the native enzymes are detected in dansyl-tripeptide cleavage specificity, inhibitor profile, immunological properties, sucrose gradient sedimentation, or gel filtration of ACE from the two tissues. However, lung and brain ACE can be differentiated in their ability to cleave amidated peptides. Both lung and brain ACE cleave Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 (substance P) via two pathways. In one pathway, ACE first releases Gly-Leu-Met-NH2 and then dipeptides sequentially from the carboxyl terminus. The other first produces Leu-Met-NH2, and then releases dipeptides to leave substance P 1-5. Lung ACE favors initial tripeptide release 3:1, while the striatal enzyme acts via the two pathways to a similar extent. Lung and striatal ACE also differ in their ability to degrade other amidated peptides. His-Lys-Thr-Asp-Ser-Phe-Val-Gly-Leu-Met-NH2 (substance K) and bombesin are degraded by striatal but not lung ACE. Physalaemin and luteinizing hormone-releasing hormone are cleaved by both enzymes, while eledoisin, kassinin, thyrotropin-releasing hormone, and substance P 5-11 are not cleaved by either enzyme. Physalaemin is degraded more rapidly by the lung enzyme. The coincidence of an ACE isozyme with substance P and substance K in the descending striatonigral pathway and the unique ability of this isozyme to cleave substance P and substance K suggest that one or both of these peptides is a physiological substrate for striatonigral ACE. PMID- 2991267 TI - Auxin-binding protein from coleoptile membranes of corn (Zea mays L.). II. Localization of a putative auxin receptor. AB - With the aid of affinity chromatography on auxin-binding protein-Sepharose (ABP Sepharose) monospecific IgGanti-ABP from rabbit antisera were isolated as judged by immuno-double diffusion test and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. With this IgGanti-ABP the ABP is localized within the outer epidermal cells of coleoptiles using indirect immunofluorescence labeling. Auxin induced growth of coleoptile segments can be inhibited by IgGanti-ABP, and the auxin response of split coleoptile sections is also strongly reduced by IgGanti ABP. The ABP, therefore, is referred to as an auxin receptor. This auxin receptor is localized at the plasmalemma of the outer epidermal cells of the coleoptile. PMID- 2991268 TI - Amplification and purification of UvrA, UvrB, and UvrC proteins of Escherichia coli. AB - The UvrA, UvrB, and UvrC proteins of Escherichia coli are subunits of a DNA repair enzyme, ABC exci nuclease. In order to amplify these proteins, we have joined the artificial canonical promoter tac (Amann E., Brosius, J., and Ptashne, M. (1983) Gene (Amst.) 25, 167-178) to the uvr genes to obtain plasmids that express these genes under the control of the lac repressor. When cells carrying the tac-uvr plasmids are induced by the gratuitous lac inducer isopropyl-beta-D galactoside the Uvr proteins are overproduced reaching a level of 10-20% of total cellular proteins after 6-8 h of induction. We have developed methods to purify all three Uvr proteins, UvrA, UvrB, and UvrC, in milligram quantities and to near homogeneity from these overproducing cells. The purified UvrA protein is an ATPase but UvrB and UvrC proteins are not. However, UvrB protein stimulates the ATPase activity of UvrA protein by a factor of 1.5 in the presence of double stranded DNA and by a factor of about 2.6 in the presence of UV-irradiated DNA but not in the absence of DNA. PMID- 2991270 TI - Site-specific phosphorylation of avian retrovirus nucleocapsid protein pp12 regulates binding to viral RNA. Evidence for different protein conformations. AB - Phosphorylation of serine 40 of the major nucleocapsid protein of avian retroviruses, pp12, regulates binding to viral RNA (Leis, J., Johnson, S., Collins, L. S., and Traugh, J. A. (1984) J. Biol. Chem. 259, 7726-7732). The phosphorylation state of the protein can be altered in vitro, resulting in the interconversion of the protein between a state of high affinity for single stranded RNA and low affinity for single- or double-stranded RNA. The reversible phosphorylation of serine 40 is accompanied by a change in the conformation of the protein as demonstrated by quenching of intrinsic tryptophan fluorescence and chemical modification studies. Quenching of fluorescence of the sole tryptophan residue, Trp 80, by poly(U), KI, and CsCl indicates that the microenvironment of this residue is more positive in pp12 than in p12. Chemical modification studies indicate that the 3 lysine residues at positions 36, 37, and 39 of pp12 react with 2,4,6-trinitrobenzenesulfonic acid, while only 1 of these residues reacts in p12. The addition of single-stranded, but not double-stranded RNA, to pp12 protects 2 of the 3 lysine residues from chemical modification, suggesting that the two protected lysyl groups are required for binding to single-stranded viral RNA. In contrast to the phosphorylation of serine 40, phosphorylation of serine 43, catalyzed by protease-activated kinase II in vitro, does not induce changes in the protein conformation nor does it alter the RNA binding properties of the protein. PMID- 2991269 TI - Characterization of an RNA polymerase I-dependent promoter within the spacer region of yeast ribosomal cistrons. AB - Nucleotide sequences which are required for RNA polymerase I-dependent selective initiation of transcription in vitro from a site within the spacer region of cloned yeast ribosomal DNA have been identified. Yeast rDNA templates containing deletion mutations extending from restriction endonuclease cleavage sites located upstream and downstream from the transcriptional initiation site were constructed. The ability of these mutant templates to support selective transcription in vitro was determined using a yeast whole cell extract. Nucleotide sequences which are required for selective transcription in vitro are within a 22-base pair region which is located immediately adjacent to the transcriptional initiation site. The 3' boundary of this 22-base pair sequence was mapped within a single base pair and resides within the transcribed portion of the rDNA. Nucleotide sequences upstream and downstream from the 22-base pair region are not required for selective transcription and do not appear to affect the efficiency of transcription in vitro. A hybrid plasmid containing only 32 base pairs of yeast rDNA, which includes the 22-base pair region, supports efficient and accurate RNA polymerase I-dependent transcription in vitro. These data demonstrate that the 22-base pair region of yeast rDNA is sufficient for accurate initiation of transcription in vitro. The transcriptional properties of several cloned rDNA templates isolated from two haploid yeast strains and a strain of bakers' yeast were examined. Four cistrons were identified which differ in nucleotide sequence. Three cistrons contain the 22-base pair promoter region and they support selective transcription in vitro. The fourth cistron does not support selective transcription in vitro and contains a single base pair substitution within the 22-base pair promoter sequence. PMID- 2991271 TI - Regulation of galactokinase gene expression in Tetrahymena thermophila. II. Identification of 3,4-dihydroxyphenylalanine as a primary effector of adrenergic control of galactokinase expression. AB - Intracellular concentrations of catecholamines were determined in wild-type and mutant Tetrahymena thermophila, using the highly sensitive techniques of high performance liquid chromatography and electro-chemical detection. Catecholamines were determined in these cell strains grown under various steady-state conditions, including those which initiate and maintain repression of galactokinase gene expression. Wild-type cells grown in defined minimal medium supplemented with 1% glycerol, exhibiting derepressed galactokinase synthesis, were found to contain considerable quantities of dopa (3,4 dihydroxyphenylalanine) and dopamine, but no detectable levels of either norepinephrine or epinephrine. Analyses of wild-type cells revealed a strong positive correlation between the internal concentration of dopa and expression of the galactokinase gene, both of which are regulated by exogenous carbohydrates, catecholamine agonists, or dibutyryl-cAMP; an analogous relationship between intracellular dopamine concentrations and galactokinase activity was not found. In addition, a correlation between intracellular dopa content and the phenotypic expression of galactokinase in various mutants deficient in the catecholamine biosynthetic pathway or in glucokinase further confirms the role of dopa as a primary effector in the regulation of galactokinase gene expression. PMID- 2991272 TI - Primary structure of histidine-tRNA synthetase and characterization of hisS transcripts. AB - Histidine-tRNA synthetase is one of the smallest bacterial aminoacyl-tRNA synthetases. It is less than one-half the size of the largest aminoacyl-tRNA synthetases. The entire nucleotide sequence of the Escherichia coli hisS locus was determined. The coding region is comprised of 424 codons, and the sequence was determined for 200 nucleotides on the 5'- and 3'-sides of the coding region. The translated nucleotide sequence was confirmed extensively by independent amino acid sequence information obtained by Edman degradations of purified peptides and by measurements of peptide masses by fast atom bombardment mass spectrometry. A significant sequence alignment of four bacterial aminoacyl-tRNA synthetases was reported recently (Webster, T., Tsai, H., Kula, M., Mackie, G., and Schimmel, P. (1984) Science 226, 1315-1317). Although the four enzymes vary considerably in length, this match occurs within the first 100 amino acids of each of the four enzymes and is in the segment believed to be part of the catalytic core. But no strong alignment could be found of the histidine sequence with these four tRNA synthetase sequences. This enzyme may be derived, therefore, from a different progenitor. Previous work suggested that three places in the hisS 5'-noncoding sequence could be promoter sites for RNA polymerase (Eisenbeis, S. J., and Parker, J. (1982) Gene 18, 107-114). We detected a 1400-nucleotide RNA species by RNA blot analysis with a hisS-specific probe. S1 nuclease mapping demonstrated a 5'-end to the RNA species occurs at -67 +/- 1, relative to the first nucleotide of the coding region. This position coincides with the predicted start site for transcription from one of the previously proposed promoter sites. PMID- 2991273 TI - Insulin affects the sodium affinity of the rat adipocyte (Na+,K+)-ATPase. AB - The K0.5 for intracellular sodium of the two forms of (Na+,K+)-ATPase which exist in rat adipocytes (Lytton, J., Lin, J. C., and Guidotti, G. (1985) J. Biol. Chem. 260, 1177-1184) has been determined by incubating the cells in the absence of potassium in buffers of varying sodium concentration; these conditions shut off the Na+ pump and allow sodium to equilibrate into the cell. The activity of Na+,K+)-ATPase was then monitored with 86Rb+/K+ pumping which was initiated by adding isotope and KCl to 5 mM, followed by a 3-min uptake period. Atomic absorption and 22Na+ tracer equilibration were used to determine the actual intracellular [Na+] under the different conditions. The K0.5 values thus obtained were 17 mM for alpha and 52 mM for alpha(+). Insulin treatment of rat adipocytes had no effect on the intracellular [Na+] nor on the Vmax of 86Rb+/K+ pumping, but did produce a shift in the sodium ion K0.5 values to 14 mM for alpha (p less than 0.025 versus control) and 33 mM for alpha(+) (p less than 0.005 versus control). This change in affinity can explain the selective stimulation of alpha(+) by insulin under normal incubation conditions. Measurement of the K0.5 for sodium ion of (Na+,K+)-ATPase in membranes isolated from adipocytes revealed only a single component of activation with a low K0.5 of 3.5 or 12 mM in the presence of 10 or 100 mM KCl, respectively. Insulin treatment of the isolated membranes or of the cells prior to membrane separation had no effect on these values. PMID- 2991274 TI - Biochemical characterization of positive cooperativity in the binding of 1 alpha, 25-dihydroxyvitamin D3 to its chick intestinal chromatin receptor. AB - We have characterized a positive cooperativity mechanism in the binding of 1,25 dihydroxyvitamin D3 (1,25-(OH)2D3) to its chick duodenum chromatin receptor. The Hill plot which can take account of the possibility of cooperativity resulted in a much better fitting of the experimental data than the Scatchard model (r = +0.998 versus r = -0.94). Concentrating the chromatin receptor preparation from 10 to 40% resulted in an increase of the Hill coefficient (nH) from 1.09 +/- 0.08 to 1.46 +/- 0.08 (S.D.). Increasing the temperature of incubation from 1 degree C to 40 degrees C resulted in a decrease of nH from 1.46 +/- 0.08 to 1.10 +/- 0.02 (S.D.). The calculation of the thermodynamics of the interaction of 1,25-(OH)2D3 with the second binding site of the receptor (from a Van't Hoff plot) showed that this process occurred spontaneously (delta G0 = -11.6 kcal X mol-1 at 1 degree C), was entropy-driven (delta S0 = +26 cal degree-1 mol-1), and was energy requiring (delta H0 = -4.37 kcal X mol-1). The temperature controlled reversibility of the cooperativity demonstrates that this phenomenon is not an artifact. Finally, in a study of the rate of dissociation of [3H]1,25-(OH)2D3 from the duodenal receptor preparation, we have found two slopes (k-1 = 32 X 10( 3) min-1; k-2 = 3.2 X 10(-3) min-1); this suggests the existence of two species of receptor. These receptor species could result possibly from either a monomer dimer system or from a conformational change of a monomer via site-site interactions. In conclusion, the positive cooperativity in the binding of 1,25 (OH)2D3 to the two binding sites of its intestinal receptor is an entropy-driven process and requires energy, is reversible with temperature, and has been shown to take place in concentrated chromatin aggregates. PMID- 2991275 TI - Ferric iron and superoxide ions are required for the killing of cultured hepatocytes by hydrogen peroxide. Evidence for the participation of hydroxyl radicals formed by an iron-catalyzed Haber-Weiss reaction. AB - Cultured hepatocytes pretreated with the ferric iron chelator deferoxamine were resistant to the toxicity of H2O2 generated by either glucose oxidase or by the metabolism of menadione (2-methyl-1,4-naphthoquinone). Ferric, ferrous, or cupric ions restored the sensitivity of the cells to H2O2. Deferoxamine added to hepatocytes previously treated with this chelator prevented the restoration of cell killing by only ferric iron. The free radical scavengers mannitol, thiourea, benzoate, and 4-methylmercapto-2-oxobutyrate protected either native cells exposed to H2O2 or pretreated hepatocytes exposed to H2O2 and given ferric or ferrous iron. Superoxide dismutase prevented the killing of native hepatocytes by either glucose oxidase or menadione. With deferoxamine-pretreated hepatocytes, superoxide dismutase prevented the cell killing dependent upon the addition of ferric but not ferrous iron. Catalase prevented the killing by menadione of deferoxamine-pretreated hepatocytes given either ferric or ferrous iron. Deferoxamine pretreatment did not prevent the toxicity of t-butyl hydroperoxide but did, however, prevent that of cumene hydroperoxide. It is concluded that both ferric iron and superoxide ions are required for the killing of cultured hepatocytes by H2O2. The toxicity of H2O2 is also dependent upon its reaction with ferrous iron to form hydroxyl radicals by the Fenton reaction. The ferrous iron needed for this reaction is formed by the reduction of cellular ferric iron by superoxide ions. Such a sequence corresponds to the so-called iron-catalyzed Haber-Weiss reaction, and the present report documents its participation in the killing of intact hepatocytes by H2O2. Cumene hydroperoxide but not t-butyl hydroperoxide closely models the toxicity of hydrogen peroxide. PMID- 2991276 TI - Reduced levels of cardiac cAMP-dependent protein kinase in spontaneously hypertensive rat. AB - Cardiac cAMP-dependent protein kinases were compared between the spontaneously hypertensive rat and the age-matched normotensive Wistar-Kyoto rat by DEAE cellulose chromatography, photoaffinity labeling with 8-N3[32P]cAMP, and Western blots using the antiregulatory and 125I-anticatalytic subunit antibodies. DEAE cellulose chromatography revealed that the ratio of type I to type II cAMP dependent protein kinase was 3:1 in the cytoplasmic soluble proteins from the heart of normotensive rat. In contrast, the ratio of type I to type II was 1:1 in the heart of hypertensive rat. Type I protein kinase was reduced by 3-fold in hypertensive rat compared to normotensive rat. The levels of type II protein kinase were similar in both normotensive and hypertensive rats. The ratio of regulatory subunits of type I (RI) to type II (RII) cAMP-dependent protein kinase was 2.5 in the soluble proteins from the heart of normotensive rat compared to a ratio of 0.62 for hypertensive rat. RI was reduced by 4-fold in hypertensive rat compared to normotensive rat. The decrease in RI from hypertensive rat was also demonstrated by photoaffinity labeling with 8-N3[32P] cAMP. Western blot analysis of the catalytic subunit revealed a 2-fold decrease in catalytic subunit (C) in the soluble proteins from the hypertensive rat compared to normotensive rat. These results show that the reduced level of activity of cardiac type I protein kinase in hypertensive rat was the result of a decrease in both the RI and C subunits, thus reducing the number of type I cAMP-dependent protein kinase holoenzyme molecules. Comparison of type I protein kinase from "prehypertensive" and "hypertensive" stages of hypertensive rat indicated that the type I protein kinase was reduced by 3-fold before an increase in the blood pressure was detectable. Cardiac type I protein kinase is predominantly associated with the cytoplasmic proteins in both the normotensive and hypertensive rats. The levels of RI, RII, and C associated with the membrane-solubilized proteins were not affected in the hypertensive rat. The levels of RII were similar in the brain tissue of normotensive and hypertensive rats, suggesting that the decrease in type I protein kinase is specific in hypertensive rat. In conclusion, a decrease in cardiac type I cAMP-dependent protein kinase may affect the degree of phosphorylation of cardiac regulatory proteins, thus impairing normal cardiac physiology in hypertensive rat. PMID- 2991277 TI - Multiple forms of the CheB methylesterase in bacterial chemosensing. AB - The methylesterase which catalyzes demethylation of chemotactic membrane receptors in Salmonella typhimurium has been purified and characterized. Two forms of the enzyme have been isolated from cell extracts. One corresponds in molecular weight, Mr = 37,000, and amino acid composition to the predicted product of the structural gene for the methylesterase, cheB. The other is a proteolytic fragment, Mr = 21,000, corresponding to the C-terminal three-fifths of the intact CheB protein. The specific activity of the 21-kDa enzyme is at least 15-fold greater than that of its 37-kDa precursor. We conclude that the CheB protein is composed of at least two structurally distinct portions: a C terminal catalytic domain, and an N-terminal region which modulates esterase activity. PMID- 2991278 TI - Solubilization and reconstitution of the gastric H,K-ATPase. AB - Proteoliposomes containing the hog gastric H+,K+-ATPase were prepared from cholate and n-octyl glucoside extracts of native microsomes. Experiments were presented which show reconstitution-dependent selective purification of a 94-kDa peptide capable of Rb+/Rb+ exchange and active H+ transport. The absence of selective enrichment of residual protein contamination in this material suggests but does not prove that those transport reactions are attributable only to the 94 kDa peptide. Transport demonstrated inhibitor sensitivity and cation specificity comparable to the microsomal gastric ATPase. In K2SO4 media the H+ transport reaction was protonophore insensitive and correlated with MgATP-dependent 86Rb+ extrusion. This and other evidence suggested that active transport occurs via electroneutral H+in for K+out exchange. 86Rb+ exchange (uptake) in the proteoliposomes demonstrated both saturable and nonsaturable components. At a K0.5 = 1.5 mM, saturable 86Rb+ uptake accounted for about 90% of Rb+ influx. The vanadate-sensitive cation exchange indicated that the ATPase was reconstituted asymmetrically into the proteoliposomes (70% cis-/30% trans-vanadate site). 86Rb+ exchange was inhibited by ATP and stimulated about 2-fold by low Mg2+ and 5 mM phosphate. These ligand effects and the demonstration of comparable rates of passive exchange and active Rb+ efflux suggest that passive K+ exchange is not severely limited by a K+-occluded enzyme form in the H,K-ATPase. A model compatible with this hypothesis is suggested. PMID- 2991279 TI - The human vasopressin gene is linked to the oxytocin gene and is selectively expressed in a cultured lung cancer cell line. AB - The human genes for prepro-arginine-vasopressin-neurophysin II (prepro-AVP-NPII) and prepro-oxytocin-neurophysin I (prepro-OT-NPI) were cloned from a human genomic library and the nucleotide sequence of both genes was determined. The two genes are similar in their intron-exon structure, linked together with 12 kilobases intervening, and transcribed from opposite DNA strands. A human small cell lung cancer cell line, H378, produces significant quantities of pre-pro-AVP NPII mRNA using a transcription unit predicted from the genomic DNA sequence. Despite the proximity of the actively transcribed prepro-AVP-NPII gene, transcription of prepro-OT-NPI is not detected in this cell line. PMID- 2991280 TI - Uridine kinase from Ehrlich ascites carcinoma. Purification and properties of homogeneous enzyme. AB - Uridine kinase from Ehrlich ascites tumor cells has been purified about 60,000 fold to apparent homogeneity and with an overall recovery of about 40%. This purification was achieved using phosphocellulose and adenosine 5'-triphosphate agarose affinity chromatography. The subunit molecular mass as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 31,000 daltons. With two dimensional electrophoresis, only one spot was observed, indicating the absence of isoenzymes. Multiple peaks of activity are routinely observed on ion exchange chromatography or gel filtration, for both crude preparations or homogeneous uridine kinase, in agreement with our earlier results that this enzyme exists as multiple interconvertible oligomeric forms (Payne, R. C., and Traut, T. W. (1982) J. Biol. Chem. 257, 12485-12488). The purified enzyme has a specific activity of 283 mumol/min/mg of protein at 22 degrees C. Initial velocity studies using uridine and ATP are consistent with a sequential mechanism. Km values for uridine, cytidine, and ATP are 40, 57, and 450 microM, respectively. CTP and UTP are competitive inhibitors with respect to ATP, with Ki values for CTP and UTP of 10 and 61 microM, respectively. The enzyme was active with several nucleoside analogs, the Km values being 69 microM (5-fluorouridine), 200 microM (3 deazauridine), and 340 microM (6-azauridine). The pure enzyme is very sensitive to freezing, but can be maintained at O degrees C for 8 weeks with only 20% loss of activity. For long-term storage, enzyme in 50% glycerol can be maintained at 20 degrees C for many months with no detectable loss of activity. PMID- 2991281 TI - Human 3-hydroxy-3-methylglutaryl coenzyme A reductase. Conserved domains responsible for catalytic activity and sterol-regulated degradation. AB - A full length cDNA for human 3-hydroxy-3-methylglutaryl coenzyme A reductase, the membrane-bound glycoprotein that regulates cholesterol synthesis, was isolated from a human fetal adrenal cDNA library. The nucleotide sequence of this cDNA shows that the human reductase is 888 amino acids long and shares a high degree of homology with the hamster enzyme. The amino-terminal membrane-bound domain is the most conserved region between the two species (7 substitutions out of 339 amino acids). This region, which is predicted to span the endoplasmic reticulum membrane seven times, mediates accelerated degradation of reductase in the presence of sterols. The carboxyl-terminal catalytic domain is also highly conserved (22 substitutions out of 439 amino acids). However, the linker region between these two domains has diverged (32 substitutions out of 110 amino acids). Conservation of the structure of the membrane-bound domain in HMG-CoA reductase supports the hypothesis that sterol-regulated degradation is an important mechanism for suppression of reductase activity and for regulation of cholesterol metabolism in humans as well as in hamsters. PMID- 2991282 TI - Photoaffinity labeling of an antimycin-binding site in Rhodopseudomonas sphaeroides. AB - Tritium-labeled 3-azidosalicyl-N-(n-octadecyl)amide was synthesized and used as a photoaffinity probe for the antimycin-binding site in both purified ubiquinone cytochrome b-c1 oxidoreductase and chromatophore vesicles from the photosynthetic bacterium Rhodopseudomonas sphaeroides. In both systems, a prominently labeled protein had a molecular weight of 11,000. Binding to this protein was inhibited by preincubation of the reaction mixture with antimycin prior to addition of the radioactive analog and subsequent irradiation. The antimycin analog, 3 azidosalicyl-N-(n-octadecyl)amide, inhibited succinate-cytochrome c reductase activity in chromatophore vesicles by 50% at a concentration of 150 nmols/mg of protein. PMID- 2991283 TI - Epidermal growth factor-mediated activation of an S6 kinase in Swiss mouse 3T3 cells. AB - Extracts from epidermal growth factor (EGF)-stimulated Swiss mouse 3T3 cells are up to 10 times more potent in phosphorylating ribosomal protein S6 than extracts from quiescent cells. Preparation of extracts in the absence of phosphatase inhibitors leads to a time-dependent loss of kinase activity. In order of potency, the most efficient phosphatase inhibitors in protecting the S6 kinase activity are phosphotyrosine followed by p-nitrophenyl phosphate, beta-glycerol phosphate, and phosphoserine. The kinetics of kinase activation following EGF treatment are rapid and transient. The maximum increase is observed between 15 and 30 min with only 20-30% of the activity remaining after 2 h. Phosphorylation of S6 in the intact cell follows a similar pattern of activation, reaching a maximum between 30 and 60 min and then slowly returning to basal levels by approximately 3 h. The activation of protein synthesis is also rapid; however, in contrast to the transient activation of the S6 kinase and S6 phosphorylation, it remains persistently high for at least 6 h following EGF treatment. Comparison of these events with EGF binding shows that about 50% of the cell surface binding sites are lost within 10 min of exposure to EGF, and about 25% remain after 2 h. Finally, sodium orthovanadate, which is known to mimic the mitogenic effect of EGF, also leads to activation of the S6 kinase, however, with distinct kinetics and by an apparent EGF receptor-independent pathway. PMID- 2991284 TI - Relationship between the structure of chromosomal protein HMG1 and its accumulation in the cell nucleus. AB - When microinjected into the cytoplasm of cultured mammalian cells, non-histone chromosomal protein HMG1 migrates into the nucleus and binds to the chromatin. To define the features of the HMG1 molecule which are essential for this activity, fragments of HMG1 and chemically modified HMG1 molecules were injected into HeLa cells and the capacity of each of these probes to accumulate in the nucleus was measured by an autoradiographic technique. Fragments representing the C-terminal and central portions of HMG1 did not concentrate in the nucleus; a fragment which consisted of the N-terminal two-thirds of the molecule and which lacked the 41 consecutive aspartate and glutamate residues located near the C-terminal end of the molecule accumulated to about the same extent as intact HMG1. When the amino groups of HMG1 were chemically modified, there was a progressive loss in the ability of the protein to accumulate in the nucleus; derivatization of one-fourth of the total amino groups reduced the concentration of microinjected protein in the nucleus relative to that in the cytoplasm to one-half of the original value. In contrast, modification of one-fourth of the total carboxyl groups did not significantly affect the capacity of HMG1 to accumulate in the nucleus, although further modification resulted in decreased nuclear accumulation. Iodination of tyrosine residues was without effect and modification of the cysteine residues had only a modest effect on the ability of HMG1 to concentrate in the nucleus. PMID- 2991285 TI - Multiple mRNAs for 3-hydroxy-3-methylglutaryl coenzyme A reductase determined by multiple transcription initiation sites and intron splicing sites in the 5' untranslated region. AB - The current studies show that mRNAs with 16 different 5'-untranslated regions (varying in length from 68 to 670 nucleotides) are produced from the 3-hydroxy-3 methylglutaryl coenzyme A reductase gene in hamster UT-1 cells. This complex pattern of mRNAs results from a combination of multiple transcription initiation sites and multiple 5' splice donor sites for the intron in the 5'-untranslated region of the gene. Analysis of the multiple mRNAs was made possible by a modification of the S1 nuclease technique in which we used a series of progressively truncated uniformly labeled, single-stranded [32P]DNA probes in addition to the usual end-labeled 32P-probes. All of the reductase mRNAs are diminished when UT-1 cells are incubated with sterols, indicating that all of them are subject to coordinate control. PMID- 2991286 TI - Regulation of Escherichia coli purF. Analysis of the control region of a pur regulon gene. AB - Escherichia coli purF has been determined to be the distal gene of a polycistronic operon. The first gene of the purF operon encodes a hydrophobic 17.9-kDa protein of unknown function. Deletion analyses indicate that the 17.9 kDa protein plays no role in the regulation of purF in cis. mRNA hybridization studies establish that purF is regulated at the transcriptional level. Enzyme and mRNA levels are repressed 11-17-fold by excess adenine. A single mRNA start site at nucleotide +1 was identified for transcripts synthesized in vivo. Two sites, at +1 and approximately +30, were used for transcription initiation in vitro. The purF promoter is localized between nucleotides -96 and -7 with sequences upstream of -71 necessary for high level expression. Initial evidence suggests that transcription is subject to stringent control. Deletion analyses localize the purF control element to a region between nucleotides -71 and +35. A putative control site between nucleotides -35 to +3 strongly resembles a 5' flanking sequence in the co-regulated gene purM. This site contains an imperfect inverted repeat sequence that is characteristic of sites recognized by regulatory proteins and is a candidate for the purF operator. This is the first detailed analysis of a gene involved in de novo purine nucleotide biosynthesis. PMID- 2991287 TI - Dephosphorylation of myosin by the catalytic subunit of a type-2 phosphatase produces relaxation of chemically skinned uterine smooth muscle. AB - It is now well-established that phosphorylation of the 20,000-dalton light chain of smooth muscle myosin (LC20) is a prerequisite for muscle contraction. However, the relationship between myosin dephosphorylation and muscle relaxation remains controversial. In the present study, we utilized a highly purified catalytic subunit of a type-2, skeletal muscle phosphoprotein phosphatase (protein phosphatase 2A) and a glycerinated smooth muscle preparation to determine if myosin dephosphorylation, in the presence of saturating calcium and calmodulin, would cause relaxation of contracted uterine smooth muscle. Addition of the phosphatase catalytic subunit (0.28 microM) to the muscle bath produced complete relaxation of the muscle. The phosphatase-induced relaxation could be reversed by adding to the muscle bath either purified, thiophosphorylated, chicken gizzard 20,000-dalton myosin light chains or purified, chicken gizzard myosin light chain kinase. Incubation of skinned muscles with adenosine 5'-O-(thiotriphosphate) prior to the addition of phosphatase resulted in the incorporation of 0.93 mol of PO4/mol of LC20 and prevented phosphatase-induced relaxation. Under all of the above conditions, changes in steady-state isometric force were associated with parallel changes in myosin light chain phosphorylation over a range of phosphorylation extending from 0.01 to 0.97 mol of PO4/mol of LC20. We found no evidence that dephosphorylation of contracted uterine smooth muscles, in the presence of calcium and calmodulin, could produce a latch-state where isometric force was maintained in the absence of myosin light chain phosphorylation. These results show that phosphorylation or dephosphorylation of the 20,000-dalton myosin light chain is adequate for the regulation of contraction or relaxation, respectively, in glycerinated uterine smooth muscle. PMID- 2991288 TI - The subunit structure of the follitropin receptor. Chemical cross-linking of the solubilized follitropin-receptor complex. AB - Homobifunctional cross-linkers were utilized to characterize high affinity (Ka = 2.2 X 10(-10) M-1) follitropin (FSH) receptors in immature bovine testis. Following the formation of radioiodinated human FSH (125I-hFSH)-receptor complexes, the membranes were solubilized with Triton X-100 or beta-octyl glucoside and the supernatants from ultracentrifugation (220,000 X g) subjected to gel filtration (Sephadex G-200) to separate the labeled hormone-receptor complexes from the unbound 125I-hFSH. The appearance of a high molecular weight (greater than or equal to 200,000) radioactive component in the elution profile was abolished when an excess of unlabeled hFSH was included in the initial incubation. After concentration by ultrafiltration, the 125I-hFSH-receptor complex, as well as the free hormone, was treated with a variety of chemical cross-linkers and subjected to analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Bands of Mr = 65,000 and 83,000 observed in the autoradiograph of the hormone-receptor complex was not present in autoradiographs of free 125I-hFSH, nor were they present when an excess of unlabeled hFSH was included in the initial binding incubation mixtures. The 65,000 and 83,000 Mr bands were, therefore, considered to represent cross-linked complexes of labeled hFSH (Mr = 38,000) or its subunits (hFSH alpha, Mr = 16,000; hFSH beta, Mr = 21,000) and components of the FSH receptor. The bands were observed on autoradiographs when the extraction of the membranes was performed with either Triton X-100 or beta-octyl glucoside and when cross-linking was accomplished with disuccinimidyl suberate, ethylene glycol bis(succinimidyl succinate), or bis[2-(succinimido oxycarbonyl)oxyethyl]sulfone. The Mr of the native FSH receptor in the calf testis has been estimated at 146,000. Our studies demonstrate the multimeric nature of the FSH receptor. However, FSH is also composed of subunits, so that due to the complexity of the system, it was not possible to arrive at a precise assessment of the Mr or quaternary structure of the receptor subunits. PMID- 2991289 TI - Effects of altered thyroid status on beta-adrenergic actions on skeletal muscle glycogen metabolism. AB - The effects of hypothyroidism on glycogen metabolism in rat skeletal muscle were studied using the perfused rat hindlimb preparation. Three weeks after propylthiouracil treatment, serum thyroxine was undetectable and muscle glycogen and Glc-6-P were decreased. Basal and epinephrine-stimulated phosphorylase a and phosphorylase b kinase activities were also significantly reduced, as were epinephrine-stimulated cAMP accumulation and cAMP-dependent protein kinase activity. Conversely, basal and epinephrine-stimulated glycogen synthase I activities were significantly higher while the Ka of the enzyme for Glc-6-P was lower in hypothyroid animals. Propylthiouracil-treated rats also had increased phosphoprotein phosphatase activities towards phosphorylase and glycogen synthase and decreased activity of phosphatase inhibitor 1. beta-Adrenergic receptor binding and basal and epinephrine-stimulated adenylate cyclase activities were reduced in muscle particulate fractions from hypothyroid rats. Administration of triiodothyronine to rats for 3 days after 3 weeks of propylthiouracil treatment restored the altered metabolic parameters to normal. It is proposed that the decreased beta-adrenergic responsiveness of the enzymes of glycogen metabolism in hypothyroid rat skeletal muscle is due to increased activity of phosphoprotein phosphatases and to reduced beta-adrenergic receptors and adenylate cyclase activity. PMID- 2991291 TI - Dual effect of GABA on the contractile activity of the guinea-pig isolated urinary bladder. AB - The effects of GABA and related substances were examined in isolated detrusor strips from the dome of the guinea-pig urinary bladder. GABA (0.01-1 mM) produced concentration-related phasic contractions of isolated strips from the guinea-pig urinary bladder dome. This effect of GABA was mimicked by homotaurine and muscimol, selective GABAA receptor agonists but not by (+/-)-baclofen, a selective GABAB receptor agonist. A specific cross desensitization was observed between GABA, homotaurine and muscimol but not between (+/-)-baclofen and GABA. GABA (1 mM)-induced contractions were antagonized by picrotoxin, a selective GABAA receptor antagonist. GABA-induced contractions were almost abolished by tetrodotoxin (0.5 microM, TTX) thus indicating their neurogenic origin. In addition GABA-induced contractions were partially antagonized by atropine (to about the same extent as those produced by dimethylphenylpiperazinium (DMPP), a ganglionic stimulant), but were unaffected by hexamethonium (10 microM), phentolamine (0.2 microM) or indomethacin (5 microM). In the presence of GABA the contractile effect of both DMPP (TTX-sensitive) and acetylcholine (ACh, TTX insensitive) were significantly reduced. Similar findings were obtained with DMPP, i.e. in preparations exposed to this ganglionic stimulant both GABA- and ACh-induced contractions were depressed. Homotaurine but not (+/-)-baclofen mimicked the depressant effect of GABA on DMPP-induced contractions. The depressant effect of GABA on ACh-induced contractions of the guinea-pig urinary bladder was neurogenic in origin, i.e., was not observed in preparations exposed to TTX. These experiments indicate that GABA has a dual effect on the contractile behaviour of the guinea-pig isolated urinary bladder. Recently it has been proposed that endogenous GABA plays a neuromodulatory role in this organ. Our data suggest that in the early phase of neurogenic activation of detrusor muscle (micturition reflex) GABA might transiently enhance excitatory neurotransmission followed by a more sustained inhibition of contractility. PMID- 2991290 TI - Effects of bromocriptine on catecholamine receptors mediating cardiovascular responses in the pithed rat. AB - The interaction of bromocriptine with several catecholamine receptors that control the sympathetic responses at cardiac and vascular level has been studied in pithed adrenalectomized and vagotomized normotensive rats. Bromocriptine (30 and 100 micrograms/kg) inhibited the stimulation-induced pressor responses in the pithed rat without modifying the pressor responses induced by noradrenaline. Sulpiride (0.3 mg/kg) abolished the effects of bromocriptine (30 micrograms/kg) but only partially prevented the effects of bromocriptine (100 micrograms/kg) on the stimulation-induced pressor responses. Yohimbine (0.3 mg/kg) partially antagonised the inhibitory effect of bromocriptine on stimulation-induced pressor responses. Combination of yohimbine and sulpiride abolished attenuation of the stimulation-induced pressor responses by bromocriptine (100 micrograms/kg). Bromocriptine (0.3 and 1 mg/kg) shifted to the right the frequency-response curve of increases in heart rate. This effect was prevented by yohimbine (0.3 mg/kg) but not by sulpiride (0.3 mg/kg). The same doses of bromocriptine were ineffective on heart rate increases induced by noradrenaline. Bromocriptine (0.3 and 1 mg/kg) shifted to the right the increases in diastolic blood pressure induced by methoxamine without modifying those induced by xylazine and noradrenaline. These results suggest that bromocriptine acts on the peripheral sympathetic nervous system of the pithed rat as an agonist of presynaptic dopamine receptors and alpha 2-adrenoreceptors and as an antagonist of postsynaptic alpha 1-adrenoreceptors. PMID- 2991292 TI - Autonomic pharmacology of leukotrienes. PMID- 2991293 TI - Anti-proliferative effects of 1,2-diphenylethane oestrogens and anti-oestrogens on human breast cancer cells. AB - The anti-tumour activities of 1,2-diphenylethane oestrogens (hexoestrol and orthohexoestrol) and anti-oestrogens (metahexoestrol, tetramethylHES, and metatetramethylHES) were studied on the human MCF-7 and MDA-MB-231 breast cancer cell lines. On the E2R-positive MCF-7 cell line, all test compounds exhibited a dose-dependent inhibition of cell proliferation, but no correlation between anti proliferative activity and binding affinity for the E2R was found. Tested on the E2R-negative MDA-MB-231 cell line, metahexoestrol also showed dose-dependent inhibitory effects, but higher concentrations were necessary than on the MCF-7 cell line. From this it is concluded that the anti-proliferative effect is specific and at least partially mediated via the E2R. Combination of metahexoestrol (10(-6)M) with E2 (10(-9) to 10(-7)M) gave no rescue effect. It is therefore suggested that this compound might be useful for therapy in the presence of high oestrogen levels, i.e. in pre-menopausal patients. The test compounds (10(-8) to 10(-6)M) could rescue the inhibitory effect of tamoxifen (10(-6)M) in a dose-dependent manner, except in the cases of metahexoestrol (10( 6)M) and tetramethylHES (10(-6)M). The latter compound exhibited a strongly additive effect at this concentration. PMID- 2991294 TI - Proviral unit II of endogenous mouse mammary tumour virus is selectively amplified and expressed in C57B1/10 mammary tumours induced by non-viral carcinogens. AB - Restriction enzyme analysis and molecular hybridization assay of DNA isolated from C57B1/10 mammary adenocarcinomas induced by a combination of dimethylbenzanthracene, oestrogen, and prolactin, revealed the presence of four extra copies of endogenous mouse mammary tumour virus (MMTV). PstI restriction pattern of the amplified proviral sequences indicated their identity with the proviral Unit II of endogenous MMTV. The amplified proviruses are hypomethylated and expressed in a hormone-dependent fashion. Their internal structure is slightly modified, since an additional EcoRI recognition site is present within the proviral genomic DNA. Selective amplification of Unit II MMTV provirus in the course of mammary tumourigenesis initiated by chemical carcinogens and hormones is compatible with the accepted multifactorial nature of this process, and is interpreted in terms of the insertional mutagenesis model for MMTV-induced oncogenesis. However, sequences of cellular DNA, adjacent to the amplified Unit II proviruses, show no homology to the integration domains int-1 and int-2 common to exogenous MMTV. PMID- 2991295 TI - Transferrin receptors in cultured breast cancer cells. AB - Transferrin receptors were demonstrated on the cell membrane of breast cancer epithelial cells in primary or long-term culture. Diferric transferrin binding was saturable, specific and was not related to DNA content or clinical and histological features of the tumour. However a good correlation (p less than 0.01) was found between transferrin binding and thymidine incorporation. These results suggest the possibility of transferrin receptor measurement as a reflection of the proliferative activity of cultured breast tumour cells. PMID- 2991296 TI - Immortalization of human lymphocytes by co-cultivation with lethally irradiated autologous T-cell lines harbouring human T-cell leukaemia virus-I. AB - Human adult peripheral blood lymphocytes were successfully immortalized by co cultivation with irradiated autologous and homologous T-cell lines harbouring human T-cell leukaemia virus-I (HTLV-I). The efficiency of transformation was the same in both cases. The participation of alloantigen stimulation in co cultivation procedures is discussed, and it is stressed that factors other than alloantigen stimulation might be required for the efficient immortalization of human T-lymphocytes in vitro by HTLV. PMID- 2991298 TI - Thrombomodulin is found on endothelium of arteries, veins, capillaries, and lymphatics, and on syncytiotrophoblast of human placenta. AB - We have used antibodies to human thrombomodulin isolated from placenta to investigate the distribution of this cofactor for protein C activation in human tissues. Thrombomodulin was found on endothelial cells of arteries, veins, capillaries, and lymphatics by immunocytochemical staining using an avidin-biotin peroxidase method. Thrombomodulin was not detected on sinusoidal lining cells of liver or on postcapillary high-endothelial venules of lymph node, although the latter contained another endothelial antigen, von Willebrand factor. Other cells noted to contain thrombomodulin antigen are those of the syncytiotrophoblast in placenta. The thrombomodulin in syncytiotrophoblast was primarily on the plasma membrane surface that forms the maternal blood sinus. Syncytiotrophoblast also stained with antibodies to von Willebrand factor, which implies that these cells have multiple endothelial functions. Thrombomodulin antigen was found in all organs studied, with the notable exception of brain. PMID- 2991297 TI - Enhancement of ricin cytotoxicity in Chinese hamster ovary cells by depletion of intracellular K+: evidence for an Na+/H+ exchange system in Chinese hamster ovary cells. AB - Depletion of intracellular K+ has been reported to result in an arrest of the formation of coated pits in human fibroblasts (Larkin, J.M., M.S. Brown, J.L. Goldstein, and R.G.W. Anderson, 1983, Cell, 33:273-285). We have studied the effects of K+ depletion on the cytotoxicities of ricin, Pseudomonas exotoxin A, and diphtheria toxin in Chinese hamster ovary (CHO) cells. The cytotoxicities of ricin and Pseudomonas toxin were enhanced in K+-depleted CHO cells whereas the cytotoxicity of diphtheria toxin was reduced by K+ depletion. The effects of NH4Cl on the cytotoxicities of ricin, Pseudomonas toxin, and diphtheria toxin were found to be similar to those of K+ depletion, and there were no additive or synergistic effects on ricin cytotoxicity by NH4Cl in K+-depleted medium. The enhancement of ricin cytotoxicity by K+ depletion could be completely reversed by the addition of K+, Rb+, and partially by the addition of Cs+, before the ricin treatment, whereas Li+ was ineffective. These protective effects of K+ or Rb+ requires a functional Na+/K+ ATPase. CHO cells grown in K+-depleted media were found to contain 6.3-fold increase in intracellular Na+ level, concomitant with a 10-fold reduction in intracellular K+ level. The enhanced cytotoxicity of ricin in K+-free medium and the increased uptake of Na+ could be abolished by amiloride or amiloride analogues, which are known to be potent inhibitors of the Na+/H+ antiport system. Our results suggest that a depletion of intracellular K+ results in an influx of Na+, which is accompanied by the extrusion of H+. Consequently, there is an alkalinization of the cytosol and the ricin-containing endosomes. As a result, ricin is more efficiently released from the endosomes in-K+-depleted cells. Results from the studies of the binding, internalization, and degradation of 125I-ricin, and the kinetics of inhibition of protein synthesis by ricin in K+ depleted cells are consistent with this working hypothesis. PMID- 2991299 TI - Processing of the asparagine-linked oligosaccharides of secreted and intracellular forms of the vesicular stomatitis virus G protein: in vivo evidence of Golgi apparatus compartmentalization. AB - The structures of the asparagine-linked oligosaccharides of several variant forms of the vesicular stomatitis virus glycoprotein transiently expressed from cloned cDNAs have been determined. Glycopeptides isolated from forms of the G protein that reach the cell surface or that are secreted into the medium are virtually identical; they contain complex-type oligosaccharides whose nonreducing ends terminate in galactose and sialic acid residues. In contrast, forms of the G protein that remain intracellular possess oligosaccharides at intermediate stages in the processing pathway. One deletion mutant, delta 1473, codes for a protein that remains in the rough endoplasmic reticulum (Rose, J. K., and J. E. Bergmann, 1982, Cell, 30:753-762) and contains only high mannose-type oligosaccharides. Another mutant, delta 1554, codes for a glycoprotein that contains oligosaccharides of primarily two classes. One class is of the high mannose type and is similar to those found on the protein coded for by delta 1473. However, the major class contains biantennary and more highly branched complex-type oligosaccharides that terminate in N-acetylglucosamine rather than galactose or sialic acid residues. These data suggest that the protein coded for by delta 1554 migrates to the Golgi apparatus, but does not enter the more distal compartment(s) of the organelle which contains galactosyl- and sialyltransferases. PMID- 2991300 TI - Intracellular sorting and basolateral appearance of the G protein of vesicular stomatitis virus in Madin-Darby canine kidney cells. AB - The polarity of the surface distribution of viral glycoproteins during virus infection has been studied in the Madin-Darby canine kidney epithelial cell line on nitrocellulose filters. Using a surface radioimmunoassay on Madin-Darby canine kidney strain I cells that had been infected with vesicular stomatitis virus or with avian influenza fowl plague virus, we found that the surface G protein was 97% basolateral, whereas the fowl plague virus hemagglutinin was 88% apical. Newly synthesized, pulse-labeled vesicular stomatitis virus appeared first on the basolateral plasma membrane as measured by an immunoprecipitation assay in which the anti-G protein antibody was applied to the monolayer either from the apical or the basolateral side. Labeled G protein could be accumulated inside the cell at a late stage of transport by decreasing the temperature to 20 degrees C during the chase. Reversal to 37 degrees C led to its rapid and synchronous transport to the basolateral surface at an initial rate 61-fold greater than that of transport to the apical side. These results demonstrate that the newly synthesized G protein is transported directly to the basolateral membrane and does not pass over the apical membrane en route. Since a previous study of the surface appearance of influenza virus hemagglutinins showed that the newly synthesized hemagglutinins were inserted directly from an intracellular site into the apical membrane (Matlin, K., and K. Simons, 1984, J. Cell Biol., 99:2131-2139), we conclude that the divergence of the transport pathway for the apical and basolateral viral glycoproteins has to occur intracellularly, i.e., before reaching the cell surface. PMID- 2991301 TI - Effect of lysosomotropic amines on the secretory pathway and on the recycling of the asialoglycoprotein receptor in human hepatoma cells. AB - We studied the intracellular transport of secretory and membrane proteins in the human hepatoma cell line HepG-2 infected with vesicular stomatitis virus. Cells were pulse-labeled in the presence of [35S]methionine and chased in the presence of the lysosomotropic agent primaquine. At a concentration of 0.3 mM primaquine effectively inhibited the secretion of albumin and, to a lesser extent, that of orosomucoid and transferrin. The drug also prevented the budding of virus particles at the cell surface. The intracellular transport to the Golgi complex of the membrane protein VSV-G was not affected by primaquine as it acquires resistance to endo-beta-N-acetylglucosaminidase H at the same rate as in control cells. Addition of primaquine at various times after the initiation of the chase period indicates that the effect of primaquine occurs just before secretion. In confirmation of the biochemical data, immunocytochemical localization of albumin in cells treated with NH4Cl demonstrated that albumin accumulated in vesicles at the trans side of the Golgi complex. The effect of primaquine on secretion was also compared with its effect on receptor recycling. The dose-response characteristics of the effect of primaquine on receptor recycling are identical to those of the effects on protein secretion and virus budding. These results indicate that both processes involve the same transport mechanism, and/or that they occur via at least one identical intracellular compartment. PMID- 2991302 TI - Alterations in chromatin conformation are accompanied by reorganization of nonchromatin domains that contain U-snRNP protein p28 and nuclear protein p107. AB - The intranuclear distribution of nuclear matrix-associated protein p107 and the 28-kD Sm antigen of U-snRNPs have been studied using double-label immunofluorescence and immunoperoxidase electron microscopy. In interphase nuclei of HeLa cells, Novikoff hepatoma cells, and rat kangaroo kidney cells, p107 was confined to discrete interchromatin domains. The domains had an irregular contour, with an average diameter of 1-1.5 micron. Each domain appeared to be composed of interconnected granules. The Sm antigen colocalized and appeared concentrated in these domains but also showed some general nucleoplasmic distribution. During mitosis, the interchromatin domains disassembled such that the Sm portion redistributed to the perichromosomal and spindle regions and the p107 component redistributed throughout the mitotic cytoplasm. During anaphase, p107 assembled into discrete clusters throughout the mitotic cytoplasm. The Sm antigen was not a component of these clusters. Double-label immunofluorescence with anti-p107 and the anti-DNA tight-binding protein, AhNa1, showed that the extranuclear p107 domains assumed an interchromatin localization only after the chromosomes had decondensed. The correlation between chromosome decondensation and the occurrence of p107 within interchromatin domains was also observed during chicken erythrocyte nuclear reactivation. We propose that the discrete interchromatin domains that contain p107 and p28 may be important for processing and splicing of RNA and that their structural assembly within nuclei is sensitive to the presence of the transcriptionally active conformation of chromatin. PMID- 2991303 TI - The exocrine protein trypsinogen is targeted into the secretory granules of an endocrine cell line: studies by gene transfer. AB - The exocrine protein rat anionic trypsinogen has been expressed and is secreted from the murine anterior pituitary tumor cell line AtT-20. We examined which secretory pathway trypsinogen takes to the surface of this endocrine-derived cell line. The "constitutive" pathway externalizes proteins rapidly and in the absence of an external stimulus. In the alternate, "regulated" pathway, proteins are stored in secretory granules until the cells are stimulated to secrete with 8-Br cAMP. On the basis of indirect immunofluorescence localization, stimulation of release, and subcellular fractionation, we find that trypsinogen is targeted into the regulated secretory pathway in AtT-20 cells. In contrast, laminin, an endogenous secretory glycoprotein, is shown to be secreted constitutively. Thus it appears that the transport apparatus for the regulated secretory pathway in endocrine cells can recognize not only endocrine prohormones, but also the exocrine protein trypsinogen, which suggests that a similar sorting mechanism is used by endocrine and exocrine cells. PMID- 2991304 TI - Distribution of glycine receptors at central synapses: an immunoelectron microscopy study. AB - The distribution of receptors for a neurotransmitter was investigated cytochemically for the first time in the central nervous system, at synapses established on cells of the ventral horn of the rat cervical spinal cord. Three monoclonal antibodies (mAb's) raised against glycine receptors were used. Immunofluorescent staining already showed discontinuous labeling at the surface of neurons, and immunoenzymatic electron microscopy further revealed that the antigenic determinants were confined to the postsynaptic membrane and concentrated at the level of the synaptic complex. More specifically, one mAb directed against the receptive subunit of the oligomeric receptor recognized an epitope on the extracellular side of the plasma membrane, whereas two other mAb's bound to the cytoplasmic face. Epitopes for the last two mAb's were more accurately localized with protein A-colloidal gold, using an intermediate rabbit anti-mouse immunoglobulin serum. (a) In addition to the presence of gold particles in areas facing the presynaptic active zone (visualized with ethanolic phosphotungstic acid), the labeling extended beyond this zone for approximately 50-60 nm, which corresponds to the width of one presynaptic dense projection. (b) The distances between the mid membrane and the gold particles were different for the two mAb's (with means of 21.7 +/- 8.5 nm and 29.8 +/- 10.4 nm, respectively). The data suggest that one of the recognized epitopes is close to the plasma membrane, whereas the second protrudes into the cytoplasm. Our results indicate that the receptor is a transmembrane protein which has a restricted spatial distribution on the postsynaptic neuronal surface. PMID- 2991305 TI - Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. AB - Human eosinophil degranulation induced by the calcium ionophore A23187 was examined by transmission and scanning electron microscopy, and morphometric analysis. After incubation with A23187, eosinophil degranulation was characterized by granule movement to the cell periphery and pentalaminar membrane fusion between perigranular and plasma membranes. As adjacent granules fused they became swollen and vesiculated; their contents were released into large cytoplasmic vacuoles, which communicated extracellulary through surface pores. Extracellular release of eosinophil peroxidase without release of lactate dehydrogenase occurred after treatment with the ionophore. Morphometric analysis of the transmission electron micrographs indicated a significant reduction of cytoplasmic granules in the degranulated cells. There was a loss primarily of larger granules and alternatively an increase in the smaller-sized granules (less than 0.1 micron2), suggesting the possibility that exocytosis of mature granules is accompanied by new cytoplasmic granule formation. PMID- 2991307 TI - [Complications of surgery of the thoraco-cervico-brachial outlets]. AB - Surgery of the thoraco-cervico-brachial outlet can be associated with a number of serious complications. This study is divided into two parts: the description of 2 cases of recurrent arterial thrombosis following operations performed in other centres, an analysis of the results of a series of 70 operations associated with transient neurological sequelae, one respiratory complication secondary to a haemothorax and 9% of cases with moderate or poor results, some of which were due to errors in indications or to incomplete excision. The literature shows that 5 types of complication can be observed: arterial, neurological, complications of the surgical approach, failures due to incomplete procedures, arterial or nervous recurrences. For these reasons, the patients submitted to these operations should be very carefully selected. This type of surgery should not be presented to patients as being free of risks and always effective. The operator should be very careful before and during the operation. If complications occur, they should be treated early and completely. PMID- 2991306 TI - Proliferation of a human embryonal carcinoma-derived cell line in serum-free medium: inter-relationship between growth factor requirements and membrane receptor expression. AB - Substantial multiplication in vitro of cloned cells from a human embryonal carcinoma cell line, Tera 2, has been obtained in a basal medium (DMEM/Ham's F12,50:50, v/v) supplemented with 10 micrograms low density lipoprotein/ml, 100 micrograms high density lipoprotein/ml, 100 ng multiplication stimulating activity/ml, 100 ng insulin/ml and 1 microgram transferrin/ml. The growth rate appears to be similar to that obtained in 10% serum. Furthermore, studies on the expression of cell surface receptors revealed that cloned Tera 2 cells express high-affinity receptors for IGF-II but not for insulin. The cells also express receptors for Epidermal Growth Factor (EGF) even though the addition of EGF does not stimulate their proliferation in serum-free medium. These results suggest that the expression of specific growth factor receptors is not an absolute determinant of hormone responsiveness. PMID- 2991308 TI - Microbore reversed-phase chromatography of proteins with conventional gradient equipment for high-performance liquid chromatography. AB - Reversed-phase high-performance liquid chromatography with microbore columns (50 X 1.0 mm) was used effectively for the separation and analysis of proteins down to 1 ng at flow-rates of 0.1-0.2 ml/min. With the use of standard low-pressure gradient HPLC equipment, the peak volumes were five times smaller when compared with a conventional column at equal chromatographic efficiencies and analysis time. The sensitivity of detection was further increased by a reduction in solvent peaks, resulting in a 20-fold overall increase. PMID- 2991309 TI - Rapid and simple purification of the 125I-labeled alpha-1 adrenergic radioligand 2-[beta-(4-hydroxylphenylethyl)aminomethyl] tetralone (BE 2254) using reversed phase high-performance liquid chromatography. PMID- 2991310 TI - High-performance liquid chromatographic assay of sulbactam using pre-column reaction with 1,2,4-triazole. AB - A high-performance liquid chromatographic (HPLC) method for the determination of sulbactam in human and rat plasma and urine has been developed. Sulbactam was reacted with 1,2,4-triazole to yield a product having an ultraviolet absorption maximum at 326 nm. The product was separated using reversed-phase HPLC from the regular components of plasma and urine with an ion-pair buffer at 50 degrees C and detected at the ultraviolet maximum. The limits of accurate determination were 0.2 and 1.0 micrograms/ml in plasma and urine, respectively. The coefficients of variation of inter- and intra-assays in human plasma spiked at 4.0 micrograms/ml (n = 5) were 1.02 and 3.05%, respectively. Coexisting cefoperazone, penicillins, or the alkaline degradation product(s) of sulbactam did not interfere in the sulbactam assay. The pharmacokinetic behaviour of sulbactam and cefoperazone coadministered to rats was estimated by moment analysis. PMID- 2991311 TI - Rapid automated high-performance liquid chromatographic analysis of cyclic adenosine 3',5'-monophosphate. Synthesis in brain tissues. AB - A rapid and sensitive automated system for measuring cyclic adenosine 3',5' monophosphate (cAMP) synthesized from either radiolabelled adenine or adenosine 5'-triphosphate (ATP) in intact and broken cell tissue preparations, respectively, is described. After incubation with radiolabelled precursor, tissue samples are deproteinized and then injected directly onto a reversed-phase high performance liquid chromatographic column. The column effluent fraction which contains cAMP is collected into scintillation vials and assayed for tritium by liquid scintillation spectrometry. Since the high-performance liquid chromatographic and fraction collection procedures are automated, over fifty samples can be analyzed in duplicate in a single day. The utility of this assay is illustrated by investigations of the effects of beta-adrenergic receptor stimulation on cAMP synthesis in tissue slices prepared from rat cerebral cortex and dopamine on cAMP synthesis in striatal membrane preparations. PMID- 2991312 TI - Evaluation of methods for chemically coupling foot-and-mouth disease virus to sheep red blood cells for immunological assays. AB - Six methods of chemically coupling proteins to red blood cells were evaluated for their effectiveness in coupling foot-and-mouth disease virus (FMDV) to sheep red blood cells. The coupling agents tested were potassium periodate, 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide hydrochloride (ECDI), chromium chloride, glutaraldehyde, bis-diazotized benzidine (BDB) and N-succinimidyl 3-(2 pyridyldithio) propionate (SPDP). Of these, only the coupling methods using BDB and SPDP resulted in virus-red cell complexes that reacted with FMDV antiserum in passive hemagglutination and passive immune hemolysis assays. The BDB and SPDP methods were studied further to determine optimal coupling conditions, the kinetics of coupling and the effects of chemical couplers on viral integrity. Only the FMDV-red cell complexes formed with SPDP were suitable targets for detecting FMDV antibody producing lymphocytes in a hemolytic plaque assay. PMID- 2991314 TI - Rotavirus concentration from raw water using positively charged filters. AB - Positively charged (PC) Zeta Plus (30 S) filters and talc-Celite (TC) layers were tested for their ability to concentrate human rotavirus (strain Wa) from seeded samples of raw water (RW), collected from the Ottawa River. In the TC technique, the RW sample was preconditioned by adding Earle's balanced salt solution (1:100) and reducing the pH to 6.0 before passing it through the TC layer. Almost 6% of the input virus was lost in the filtrate. Using 1 X tryptose phosphate broth (TPB) at pH 9.0 as an eluent, 48% of the virus could be recovered from the layers. With the 30 S PC filters, no sample preconditioning was required and no virus was detected in the filtrates. In virus elution from these filters, 1 X TPB (pH 9.0), and a mixture of 6% glycine + 6% arginine (pH 9.0) gave virus recoveries of 44% and 46%, respectively. When these same two eluents were used in sequence to recover the rotavirus from 30 S PC filters, a virus recovery of about 70% was obtained. However, when 20 1 vols. of seeded raw water were concentrated using 30 S PC filters (142 mm diameter), and polyethylene glycol hydroextraction was used for second-step concentration of the eluates, only 16% of the input infectious virus could be recovered. The 30 S PC filters were then used to recover naturally occurring rotaviruses from the waters of the Ottawa River. After concentration, these viruses were detected and quantitated in MA-104 cells using an indirect immunoperoxidase staining technique. PMID- 2991313 TI - Efficiency of in situ hybridization as a function of probe size and fixation technique. AB - In an attempt to improve fixation technique for viral RNA detection by in situ hybridization, we have quantitatively compared the hybridization signal obtained when measles virus or visna virus infected cell cultures were fixed with eight different fixatives and hybridized with 35S-labeled virus-complementary DNA probes of several size ranges. Small probes (mean length, 70 bases) gave higher signals than larger probes (mean lengths 140, 350, and 780 bases) with all fixatives. This increase in signal was minimal with acetic ethanol or formalin, but was dramatic with fixatives containing glutaraldehyde; with these fixatives the signals with small probes were 6.5- to 22-fold greater than with large probes. The highest signals were obtained with periodate-lysine-paraformaldehyde glutaraldehyde (PLPG) fixed cells hybridized with small probes, and were 1.5- to 6.7-fold greater than those obtained with the commonly used fixative acetic ethanol. PLPG and other glutaraldehyde based fixatives also greatly improved the preservation of cellular morphology compared to acetic ethanol. PMID- 2991315 TI - A simple method for clonal selection of hepatitis A virus based on recovery of virus from radioimmunofocus overlays. AB - Hepatitis A virus (HAV), a non-cytopathic picornavirus, has been quantitated in cell culture by autoradiographic detection of foci of viral replication developing beneath an agarose overlay following fixation and 'staining' of the cell sheet with radiolabelled antibody (radioimmunofocus assay). Using a modification of this basic technique, a clonal variant of HM-175 strain HAV was isolated from agarose overlying individual radioimmunofoci. Virus recovered from the agarose was amplified in small volume cultures of BS-C-1 cells and identified in supernatant culture fluids by cDNA-RNA hybridization. No virus was recovered from agarose which did not overlie a focus of viral replication. This method offers a simple, yet relatively rapid and certain means of selecting clonal variants of non-plaquing viruses such as hepatitis A virus. PMID- 2991316 TI - Evidence that the beta-adrenergic system and prostaglandins stimulate renin release through different mechanisms. AB - In normal man, converting enzyme inhibition (CEI) acutely increases plasma active renin and decreases plasma inactive renin. This reciprocal relationship suggests that conversion of inactive to active renin may be important in the acute response to stimulation of renin secretion. To determine whether the beta adrenergic system or prostaglandins (PGs) participate in the acute effect of CEI on renin, we administered captopril (50 mg) alone and with either propranolol (P; 80 mg) or a PG cyclooxygenase inhibitor [PI; indomethacin (50 mg) or ibuprofen (800 mg)] to normal subjects ingesting a 25 meq/day Na diet. Supine blood pressure fell by 12 +/- 2 (+/- SE) mm Hg with CEI alone, 10 +/- 1 mm Hg with CEI plus P, and 7 +/- 1 mm Hg with CEI plus PI. Active renin rose 8-fold (P less than 0.01), with a peak at 1-2 h, after CEI and 3-fold (P less than 0.02) in response to CEI plus P or CEI plus PI. P did not block the fall in acid-activated inactive renin compared to CEI alone. The nadir of the inactive renin response to both CEI or CEI plus P occurred at 1-2 h. PI, however, prevented the fall in inactive renin. To extend this observation, we compared the effects of infusion of a vasodilator PG (PGA1; 0.6 micrograms/kg X min) and a pure beta-agonist (isoproterenol; 0.3 micrograms/kg X min). PGA1 increased active renin 2.5-fold and decreased inactive renin by 80% (both P less than 0.02), while isoproterenol increased active renin 4.1-fold, but did not significantly change inactive renin. These data suggest that the beta-adrenergic system and PGs at least acutely stimulate renin production at different steps of its biosynthesis or secretion. PMID- 2991317 TI - In vitro effect of estrogens on the peroxidase activity of human endometrium. AB - The effect of estrogen on the induction of human endometrial peroxidase was studied in vitro. When 17 beta-estradiol was added to the incubation medium, endometrial peroxidase activity was markedly increased, and the highest enzymatic activities were reached at about 12 h of incubation. The ability of estrogen and related compounds to increase peroxidase activity at a concentration of 10(-7) M was 17 beta-estradiol greater than estrone greater than 17 alpha-ethynylestradiol greater than estriol = diethylstilbestrol. Mestranol, progesterone, and testosterone had almost no activity. The induction of peroxidase by 17 beta estradiol was inhibited by actinomycin D or cycloheximide. This method may be useful for determining the direct estrogenic effect of compounds in human endometrium. PMID- 2991318 TI - The 24-hour profile of adrenocorticotropin and cortisol in major depressive illness. AB - The 24-h profile of plasma ACTH and cortisol levels was determined in 18 men suffering from major depressive illness (8 with unipolar depression and 10 with bipolar depression) as well as in 7 age-matched normal men. Blood was sampled every 15 min. The circadian variation and episodic fluctuations were analyzed for each individual profile. Both unipolar and bipolar depressed patients had higher 24-h mean cortisol levels (P less than 0.01) than normal men, but no significant difference in 24-h mean ACTH level was found. The nadir of cortisol secretion occurred almost 3 h earlier in older normal subjects and patients with unipolar depression, regardless of age, than in younger normal subjects. This shift paralleled a similar advance of the ACTH nadir. Early timing of the quiescent period of ACTH-cortisol secretion was also found in several patients with bipolar depression, but did not reach significance at the group level. The hypercortisolism in the depressed patients was associated with an increase in the magnitude, but not the number, of cortisol secretory episodes. About 90% of the cortisol pulses could be related to a concomitant ACTH pulse in normal subjects as well as in both groups of depressed patients. However, concomitant ACTH and cortisol pulses were less correlated in magnitude in depressed patients than in normal subjects. These results indicate that major depressive illness is associated with disturbances of pituitary-adrenal function. The early timing of the nadir of ACTH-cortisol secretion suggests that disorders of circadian time keeping may characterize major endogenous depression. PMID- 2991319 TI - Sensitivity of cortisol and adrenal androgens to dexamethasone suppression in hirsute women. AB - To test the hypothesis that adrenal androgen secretion is more easily suppressed than is cortisol secretion by glucocorticoids, we examined the dose-response relationship for suppression of serum dehydroepiandrosterone (DHA), DHA sulfate (DHAS), testosterone, and cortisol by dexamethasone. Nine hirsute women received daily doses of dexamethasone, starting with 0.1 mg and increasing by 0.1-mg increments every 4 weeks, until the cortisol response to ACTH was reduced to 20% or less of the response before treatment. Serum hormone levels were measured at each dexamethasone dose before and after iv administration of 25 U synthetic ACTH. Although low doses of dexamethasone caused a similar suppression of basal cortisol, DHA, and DHAS levels, ACTH-stimulated DHA levels were suppressed to a greater extent than ACTH-stimulated cortisol levels. Higher dexamethasone doses did not result in a significant difference between the degree of cortisol and adrenal androgen suppression, as near-maximal suppression occurred for all three hormones. Maximal suppression of basal testosterone levels occurred at or below the dexamethasone dose of 0.3 mg. We conclude that the adrenal androgen secretory capacity is more sensitive to suppression by dexamethasone than is the adrenal cortisol secretory capacity. Furthermore, glucocorticoid therapy for hirsutism need not achieve complete cortisol suppression to effect a major reduction in adrenal androgen levels. PMID- 2991320 TI - Effects of proopiomelanocortin peptides and angiotensin II on steroidogenesis in isolated aldosteronoma cells. AB - Cells isolated from five aldosterone-producing adenomas were used to study glucocorticoid and aldosterone production in response to ACTH, angiotensin II (A II), and peptides derived from proopiomelanocortin (POMC), viz. the 16K N terminal fragment (16K) and its derivative, gamma 3MSH and the C-terminal fragment beta-lipotropin (beta LPH) and its derivative beta-endorphin. At concentrations similar to those of ACTH and A II (10(-12)-10(-10) M), 16K, gamma 3MSH, and beta LPH selectively stimulated aldosterone production, which reached levels close to those obtained with A II. ACTH, however, was the most effective stimulant of steroidogenesis. The 16K, gamma 3MSH, and beta LPH peptides potentiated the action of ACTH, particularly in the case of aldosterone production. beta-Endorphin, whether used alone or in association with ACTH, had no effect on steroidogenesis at the dose used (10(-10) M). The principal glucocorticoid products of the adenoma cells were cortisol and corticosterone. The ratios of corticosterone to cortisol (B/F) and aldosterone to corticosterone (A/B) varied considerably from one adenoma to another, both basally and in response to ACTH. Nevertheless, within individual adenomas, the mean B/F ratio induced by ACTH [0.280 +/- 0.013 (+/- SEM)] was significantly larger than that induced by A II (0.127 +/- 0.007; P less than 0.001). By contrast, the A/B ratio in response to ACTH (0.061 +/- 0.003) was significantly smaller than that in response to A II (0.159 +/- 0.010; P less than 0.001). The values obtained with 16K (B/F, 0.106 +/- 0.010; A/B, 0.192 +/- 0.028) and gamma 3MSH (B/F, 0.122 +/- 0.012; A/B, 0.178 +/- 0.020) were close to those obtained with A II. 16K and gamma 3MSH potentiated ACTH's effect on steroidogenesis mainly by increasing the A/B ratio from 0.061 +/- 0.003 for ACTH alone to 0.100 +/- 0.008 for 16K plus ACTH (P less than 0.005) and to 0.092 +/- 0.005 for gamma 3MSH plus ACTH (P less than 0.001). The findings suggest that the stimulation of aldosterone production by 16K and gamma 3MSH in aldosteronoma cells is of the A II type and that these peptides may play a role in the genesis of primary aldosteronism. PMID- 2991321 TI - Molecular forms of glucagon-like peptide-1 in human pancreas and glucagonomas. AB - The structure of human preproglucagon, as deduced from nucleotide sequencing of the preproglucagon gene, contains two glucagon-like peptides (GLP-1 and GLP-2) in the portion C-terminal to glucagon. A rabbit antiserum was raised against synthetic GLP-1-(1-19) which had 20% cross-reactivity with synthetic GLP-1 and des-Gly37-GLP-1 amide, two possible forms of the GLP-1 whole molecule, but no significant cross-reactivity with glucagon or other pancreatic peptides. Immunocytochemistry revealed that the distribution of GLP-1-(1-19) immunoreactivity followed that of glucagon-like immunoreactivity in the normal human pancreas and in two human glucagon-secreting pancreatic tumors. Chromatography of human pancreas extracts on Sephadex G-50 gave peaks of cross reactivity at Kav values of 0.06-0.16, 0.34-0.39, 0.54-0.58 (the elution position of synthetic GLP-1), and 0.64-0.70. The concentration of immunoreactivity in the Kav 0.54-0.58 peak measured by RIA using GLP-1 or des-Gly37-GLP-1 amide as standard was 94 +/- 7 pmol/g (mean +/- SEM), while the total pancreatic glucagon content was 4.8 +/- 0.8 nmol/g. One extract of a human glucagon-secreting pancreatic tumor contained a prominent peak of GLP-1-(1-19) peptide cross reactivity with properties identical to those of GLP-1 or des-Gly37-GLP-1 amide on gel filtration and reverse phase high pressure liquid chromatography, but another tumor contained a preponderance of cross-reactive forms of greater molecular size. Pretreatment plasma from three patients with radiological and biochemical evidence of glucagon-secreting tumors contained a peak of cross reactivity with the chromatographic properties of intact GLP-1. The low concentrations of intact GLP-1 in normal pancreas compared with pancreatic glucagon concentrations suggest that the majority of the proglucagon is cleaved in a manner that does not produce GLP-1, as defined by its delimiting pairs of basic amino acid residues. PMID- 2991323 TI - Elevated production rate of 1,25-dihydroxyvitamin D in patients with absorptive hypercalciuria. AB - Previous studies of 1,25-dihydroxyvitamin D [1,25-(OH)2D] kinetics in normal subjects using the pulse injection technique have led to conflicting results, and only limited data are available concerning 1,25-(OH)2D kinetics in hypercalciuric patients. We developed an infusion equilibrium technique that measures the metabolic clearance and production rates of 1,25-(OH)2D and applied this technique in 13 normal subjects and 9 well characterized patients with absorptive hypercalciuria; all subjects were studied after 10 days on a 400-mg calcium intake. All subjects received a constant infusion of [3H]1,25-(OH)2D3 (20,000 dpm/min). Purified plasma radioactivity reached steady state levels after 15 h, and between 15 and 19 h, serial measurements of purified plasma radioactivity and endogenous 1,25-(OH)2D were made for calculation of metabolic clearance and production rates. In the 13 normal subjects, the MCR values were within a narrow range, with a mean +/- SD value of 37 +/- 6 ml/min, which, when combined with the mean steady state concentration of endogenous 1,25-(OH)2D (42 +/- 6 pg/ml), yielded a mean production rate of 2.2 +/- 0.5 micrograms/day. In the 9 patients with absorptive hypercalciuria, MCR values also were tightly clustered, with a mean of 35 +/- 4 ml/min. However, the mean endogenous steady state 1,25-(OH)2D level was significantly elevated in these patients, such that the calculated mean 1,25-(OH)2D production rate was significantly elevated at 3.4 +/- 0.5 micrograms/day. In 7 of the 9 patients with absorptive hypercalciuria, production rates exceeded the highest values found in the normal subjects. These data demonstrate disordered 1,25-(OH)2D production as opposed to metabolic clearance in the syndrome of absorptive hypercalciuria. PMID- 2991322 TI - Effects of the antiprogesterone steroid RU 486 during midluteal phase in normal women. AB - The antiprogesterone steroid RU 486 (17 beta-hydroxy-11 beta-4-dimethyl aminophenyl)17 alpha(1-propynyl)estra-4,9-dien-3-one) was given orally to 32 normally cycling women for 4 days, starting on the fourth day of the luteal phase. Uterine bleeding occurred on the third day of RU 486 administration in all 14 women treated with 100 mg/day, in 7 of the 8 women treated with 50 mg, and in 8 of 10 women receiving 25 mg/day. Premature luteal regression induced by RU 486 occurred in 8 women treated with 100 mg/day, in 3 treated with 50 mg, and in 2 receiving 25 mg/day. Plasma LH was measured every 15 min from 0800-1200 h for 5 days in 17 women. Mean LH levels decreased and pulsatile release disappeared in 7 of the 8 women treated with 100 mg, in 2 of 4 receiving 50 mg, and in 1 of 5 treated with 25 mg. RU 486 had no effect when given to 5 women with anovulatory cycles for 4 days starting on day 18 of the cycle. IN CONCLUSION: 1) RU 486, given to normally cycling women at midluteal phase, provokes uterine bleeding. 2) This effect occurs whether or not luteal regression is induced by the compound, indicating that RU 486 acts directly upon the endometrial tissue, very likely at the progesterone receptor level. 3) The drug may impair simultaneously or separately luteal function and gonadotropin secretion in a dose-dependent manner. 4) The lack of antiglucocorticosteroid activity, at the dosage of 100 mg/day, suggests that RU 486 may be useful for fertility control. PMID- 2991324 TI - High serum levels of angiotensin-converting enzyme in untreated Addison's disease. AB - Serum angiotensin-converting enzyme (ACE) in four patients with untreated Addison's disease was significantly higher [79.7 +/- 17.5 (SD) nmol min-1 ml-1] than in normal subjects (37.7 +/- 8.9 nmol min-1 ml-1, P less than 0.02). During corticosteroid hormone replacement therapy the enzyme levels returned within the normal range (44.0 +/- 7.6 nmol min-1 ml-1). In two additional patients the enzyme was assayed only while they were receiving therapy and found normal (38 and 52 nmol min-1 ml-1, respectively). In one of them, an increase ACE level (70 nmol min-1 ml-1) was found after therapy was reduced by 50%. Primary adrenal insufficiency is another disease in which ACE levels may be increased. PMID- 2991325 TI - Follicle-stimulating hormone-secreting pituitary adenomas. AB - This retrospective study concerns 40 patients with an apparently nonsecretory pituitary adenoma who were operated on during an 11-yr period from 1971 to 1981. Among them, 6 men had elevated serum FSH levels. LH levels were normal in 5 and slightly elevated in 1. Testosterone levels were low in 2 patients and within normal limits in 2 others. Sexual impotency had developed from 6 months to 1 yr before surgery in all patients. Primary hypogonadism could be eliminated on clinical grounds (recent onset of hypogonadism, previous fertility of 5 of the 6, and postoperative improvement). After transsphenoidal adenomectomy, FSH levels returned to normal values in all, and clinical recovery occurred in most patients. Tumor tissue obtained at operation stained positively for the gonadotropins, but was negative for other pituitary hormones in all patients. The most probable explanation for these findings was that the tumors were responsible for the elevated FSH secretion. This explanation is supported by the immunocytochemical identification of gonadotropin-containing cells in the tumors. We conclude that these 6 men from a series of 40 patients who presented with pituitary tumor but no GH, PRL, or ACTH hypersecretion had primary gonadotropinomas. PMID- 2991326 TI - Epidermal growth factor binding to human amnion, chorion, decidua, and placenta from mid- and term pregnancy: quantitative light microscopic autoradiographic studies. AB - All of the cell types, but not noncellular elements, found in amnion, chorion, decidua, and placenta of mid- and term pregnancy contained numerous silver grains after incubation with 1 nM [125I]epidermal growth factor ([125I]EGF). These grains completely disappeared when excess unlabeled EGF, but not unlabeled insulin, was added with [125I]EGF, suggesting the presence of specific EGF receptors. Light cells of amnion contained more EGF receptors than dark cells of amnion, and the number of light cells decreased from mid- to term pregnancy. Numerous syncytiotrophoblasts containing a greater number of receptors than darkly stained cells in connective tissue were found in chorion from midpregnancy. The chorionic syncytiotrophoblasts were considerably reduced at term. Dark cells of decidua contained more receptors than light cells of decidua, and these cell numbers did not change during pregnancy. Placental syncytiotrophoblasts contained the EGF receptors. At mid- and term pregnancy, placenta contained the highest number of EGF receptors, followed by chorion greater than decidua greater than amnion. The receptor number in all tissues was higher at mid-compared to term pregnancy. A decrease in number of cells as well as a decrease or fewer receptors per cell during pregnancy may explain the tissue and mid- vs. term pregnancy differences. The higher number of EGF receptors in amnion, chorion, decidua, and placenta at midpregnancy, at a time when the maternal and fetal blood EGF levels are at their peak, suggests that EGF may exert maximal biological effects on the feto-placental unit at midpregnancy. PMID- 2991327 TI - Successful treatment of Cushing's syndrome with the glucocorticoid antagonist RU 486. AB - A patient with Cushing's syndrome due to ectopic ACTH secretion was treated successfully with the new glucocorticoid antagonist RU 486 [17 beta-hydroxy-11 beta-(4-dimethylamino phenyl) 17 alpha-(1-propynyl)estra-4,9-dien-3-one]. This compound is a 19-nor steroid with substitutions at positions C11 and C17 which antagonizes cortisol action competitively at the receptor level. Oral RU 486 was given in increasing doses of 5, 10, 15, and 20 mg/kg . day for a 9-week period. Treatment efficacy was monitored by assessment of clinical status and by measuring several glucocorticoid-sensitive variables, including fasting blood sugar, blood sugar 120 min after oral glucose administration, and plasma concentrations of TSH, corticosteroid-binding globulin, LH, testosterone estradiol-binding globulin, and total and free testosterone. With therapy, the somatic features of Cushing's syndrome (buffalo hump, central obesity, and moon facies) ameliorated, mean arterial blood pressure normalized, suicidal depression resolved, and libido returned. All biochemical glucocorticoid-sensitive parameters normalized. No side-effects of drug toxicity were observed. We conclude that RU 486 may provide a safe, well tolerated, and effective medical treatment for hypercortisolism. PMID- 2991328 TI - Asymptomatic rotavirus infections in day care centers. AB - Rotaviruses and other enteropathogenic agents were detected in 288 (42.1%) of 684 children in day care centers of Instituto Politecnico Nacional in Mexico City. The same agents were also found in 114 (37.7%) of 302 adults directly involved in the care of the children. The study was carried out from July to December 1982 and from July 1983 to February 1984. Rotaviruses were the main enteropathogenic agents found and were detected in 169 (29.9%) of 564 children without diarrhea and in 34 (28.3%) of 120 children with diarrhea. These viruses were present in 62 (20.5%) of 302 adults without diarrhea. Of all rotavirus-positive individuals, 20% were also positive for other enteropathogens. All these observations indicate that asymptomatic rotavirus infections are not a rare event in children and that diarrhea caused by rotavirus infections is only one of the expressions of their presence. PMID- 2991329 TI - Detection of hepatitis A virus and antibody by solid-phase radioimmunoassay and enzyme-linked immunosorbent assay with monoclonal antibodies. AB - Monoclonal antibodies (K3-2F2 and K3-4C8) raised against hepatitis A virus were used to develop a solid-phase radioimmunoassay and enzyme-linked immunosorbent assay for the detection of hepatitis A virus and antibody. Assays with this pair of monoclonal antibodies were compared in parallel with similarly constructed solid-phase radioimmunoassays and enzyme-linked immunosorbent assays in which human polyclonal serum was used. The monoclonal antibody assay proved to be more sensitive for the detection of hepatitis A virus from fecal specimens as well as for anti-hepatitis A virus immunoglobulin G (IgG) and IgM in sera. PMID- 2991330 TI - Detection of cytomegalovirus antibody with latex agglutination. AB - Transfusion-acquired cytomegalovirus (CMV) infections should be prevented in seronegative immunocompromised patients by providing blood products from donors who are also seronegative. Latex agglutination was investigated as a simple and rapid method for detecting antibody against CMV. Latex beads were coated with CMV antigen, incubated for 8 min at room temperature with 25 microliter of sera, and examined for agglutination. The sensitivity and specificity of latex agglutination was compared with that of indirect hemagglutination (IHA, Cetus Corp., Emeryville, Calif.) and enzyme immunoassay (EIA) with sera from 604 random blood donors or patients. Of 327 serum samples shown to be seronegative by EIA and IHA, 327 had a latex agglutination titer of less than 1:4 (specificity, 100%). Of 236 serum samples with detectable antibody by EIA and IHA, 228 had a latex agglutination titer of 1:4 or greater (sensitivity, 97%). Plasma collected with EDTA, heparin, or citrate was satisfactory for latex agglutination. Latex agglutination results correlated quantitatively with those of EIA, and the test also detected fourfold or greater rises in antibody with paired sera from six patients with posttransfusion CMV infections. Latex agglutination is a sensitive and specific assay that is rapid and simple to perform and should be effective in selecting seronegative blood donors to prevent posttransfusion CMV infections in seronegative recipients. PMID- 2991331 TI - DNA relatedness and biochemical features of Campylobacter spp. isolated in central and South Australia. AB - Investigations of the etiology of diarrhea in patients in South Australia and the Northern Territory showed that Campylobacter spp. other than Campylobacter jejuni and C. coli were common in children. Campylobacters which were hippurate positive, nitrate negative, and susceptible to cephalothin and polymyxins were shown to be closely related to C. jejuni by DNA studies. Thermotolerant catalase negative campylobacters were also isolated. These were H2S negative and biochemically resembled the catalase-negative or weak strains found in dogs in Sweden. DNA studies showed these campylobacters to be distinct from C. sputorum subsp. sputorum and to form a homogeneous group distinct from the enteropathogenic catalase-positive campylobacters. Preliminary studies suggest that these campylobacters are related to the Swedish catalase-negative or weak strains. PMID- 2991332 TI - Diagnosis of Colorado tick fever virus infection by enzyme immunoassays for immunoglobulin M and G antibodies. AB - An immunoglobulin M (IgM) capture enzyme immunoassay technique was adapted for the detection of antibody to Colorado tick fever virus in sera from 84 individuals for whom diagnosis had been confirmed by virus isolation or neutralization test. Titers were compared with those for IgG and neutralizing antibodies in these Colorado tick fever cases. IgM antibody titers were higher than neutralizing antibody titers, but neither appeared until 1 to 2 weeks after the onset of illness. Neutralizing antibodies were detected earlier than IgM antibodies, and both were detected with greater frequency than IgG antibodies. Late-convalescent-phase sera contained both neutralizing and IgG antibodies, but IgM was all but undetectable by 2 months after onset. Although the neutralization test may remain the serological test of choice, the enzyme immunoassay for IgM antibody offers a simple and more rapid method of serodiagnosis; the enzyme immunoassay is, however, less sensitive than the neutralization test. Furthermore, because there was a sharp decline in IgM antibody after 45 days, the presence of IgM antibody in a single serum sample provides a basis for the presumptive serodiagnosis of recent Colorado tick fever virus infection. PMID- 2991333 TI - Comparison of neutralization and DNA restriction enzyme methods for typing clinical isolates of human adenovirus. AB - Sixty-five adenovirus isolates collected over a 3.5-year period were typed by both standard microneutralization techniques and restriction endonuclease digestion of viral DNA. Of the 65 isolates, 47 (72.3%) representing six adenovirus types could be typed by microneutralization. Eighteen isolates demonstrated partial neutralization with standard antisera to two or more adenovirus serotypes and thus could not be definitively typed. DNA analysis permitted typing of 64 of the 65 isolates (98.5%) (including four isolates which contained mixtures of two adenovirus types), and 12 different types were identified. Neutralization and DNA typing disagreed for five isolates, and in each case, digestion with multiple restriction endonucleases and DNA hybridization studies were consistent with the type assigned by DNA analysis. In addition, the DNA analysis method allowed the identification of genomic variants (genome types) of five adenovirus types. We conclude that typing clinical isolates of adenovirus by restriction endonuclease digestion of viral DNA can be done rapidly, provides additional epidemiological and typing information, and provides fewer ambiguous results than does typing by neutralization. PMID- 2991335 TI - A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. AB - Considerable evidence indicates that the glycoprotein (GP) IIb/IIIa complex on human platelets functions as a receptor for fibrinogen, but little is known about the mechanism of receptor "exposure." To investigate this mechanism, our previously described murine monoclonal antibody (10E5) and a new monoclonal antibody (7E3), both of which block the binding of fibrinogen to platelets and bind to GPIIb and/or GPIIIa, were radiolabeled and their rates of binding to native and ADP-activated platelets were studied. At low concentrations, 125I-10E5 bound nearly equally rapidly to both native and activated platelets, whereas 125I 7E3 bound slowly to native platelets and much more rapidly to activated platelets. This increased rate of 7E3 binding is unlikely to be due to an increase in the number of GPIIb/IIIa sites on the surface of activated platelets because: (a) the rate of 10E5 binding was unchanged; (b) the total number of surface GPIIb/IIIa sites increased by only 2-10% with activation as judged by equilibrium binding of near-saturating concentrations of 10E5 and 7E3, and (c) there was less than 1% release of platelet factor 4 with activation, indicating minimal fusion of alpha-granule membranes (a potential source of GPIIb/IIIa) with the plasma membrane. Other activators (epinephrine, thrombin, and ionophore A 23187) also increased the rate of 7E3 binding, as did digestion of platelets with chymotrypsin. Aspirin did not affect the rate of binding of 7E3, whereas apyrase, prostaglandin E1, and dibucaine all inhibited the enhancement of the 7E3-binding rate produced by ADP. These data provide evidence for an activation-dependent change in the conformation and/or microenvironment of the GPIIb/IIIa complex, and offer a method of studying the receptor exposure mechanism that does not rely on the binding of fibrinogen itself. PMID- 2991336 TI - Modulation of renal sodium-potassium-adenosine triphosphatase by aldosterone. Effect of high physiologic levels on enzyme activity in isolated rat and rabbit tubules. AB - The purpose of this study was to determine the nephron site, time course, and mechanism of mineralocorticoid action on renal tubular Na-K-ATPase in rats and rabbits, without dietary manipulation and by using the natural mineralocorticoid aldosterone. Sustained, high physiologic levels of circulating aldosterone mimicking those produced endogenously during potassium loading or sodium deprivation were provided by constant delivery of the hormone in doses of 5 or 50 micrograms/100 g body wt per 24 h, respectively, from osmotic minipumps implanted subcutaneously. In adrenal-intact rats receiving the 5-microgram dose, aldosterone levels were similar to those seen in animals fed a high K diet and produced a time-dependent increase in Na-K-ATPase activity in the cortical collecting tubule (CCT) to a level 103% higher than in controls after 7 d (2,007 +/- 178 vs. 989 +/- 72 pmol/mm per h, P less than 0.001); the enzyme activity in the proximal convoluted tubule, medullary thick ascending limb, and the inner stripe of the medullary-collecting tubule did not change significantly. The increment in CCT Na-K-ATPase was larger (142%) in animals receiving for the same period of time the 50-micrograms dose, which produced circulating aldosterone levels similar to those of sodium-deprived rats. A significant stimulation of Na K-ATPase activity was seen in the CCT of adrenalectomized rats after 24 h of treatment with either dose of the hormone, and at 12 h only in animals receiving the 50 micrograms/100 g per 24 h regimen. To determine whether the enhanced Na-K ATPase activity produced by aldosterone is due to synthesis of new enzyme units or to alteration in its kinetics, we examined the ouabain-binding capacity and the affinity for Na and K of the enzyme from CCT of rabbits treated with 5 micrograms/100 g body wt per 24 h aldosterone for 3 d. These experiments revealed a parallel increment on Na-K-ATPase activity and specific [3H]ouabain binding in aldosterone-treated rabbits, while the affinity of the enzyme for either sodium or potassium was unaltered. The results of this study indicate that high physiologic levels of aldosterone simulating those measured during K loading or Na deprivation lead to a segment-specific increase in Na-K-ATPase activity in the CCT. This effect was time-and dose-dependent and was due to an increase in the number of active enzyme units. The segmental specificity and time course of the increase in enzyme activity suggest that modulation of Na-K-ATPase by aldosterone plays a role in the chronic adaptation of the CCT to altered availability of sodium and potassium, and therefore in the homeostasis of these cations by the kidney. PMID- 2991334 TI - Vitamin D-endocrine system. PMID- 2991338 TI - Inhibition and subsequent enhancement of platelet responsiveness by prostacyclin in the rabbit. Relationship to platelet adenosine 3',5'-cyclic monophosphate. AB - Methods were developed for measuring changes in platelet sensitivity to a release inducing stimulus and in platelet cyclic AMP in fresh whole blood samples from rabbits. These techniques permitted detection of the effects of exogenous and endogenous prostacyclin on circulating platelets. In these methods, rabbit platelets were labeled in vitro by incubation with [14C]serotonin and [3H]adenine and then transfused into other rabbits. Release of platelet [14C]serotonin by a standard dose of synthetic platelet-activating factor (40 pmol/ml) and the platelet cyclic [3H]AMP levels were then measured in citrated blood from the conscious animals within 2 min of arterial puncture. Bolus intravenous injections of prostacyclin (1-10 nmol/kg) caused concentration-dependent increases in platelet cyclic AMP after 2 min, which decreased approximately 75% by 5 min, and disappeared after 30 min. Significant inhibition of the platelet release reaction was detected 2 min but not 5 min after injection of 10 nmol of prostacyclin per kilogram. With lower doses, significant enhancement of the release of [14C]serotonin was observed after 5 min. Similar changes in platelet responsiveness and cyclic [3H]AMP were observed after release of endogenous prostacyclin by intravenous injection of angiotensin II (5 nmol/kg); inhibition of the release of [14C]serotonin after 2 min was followed by potentiation after 5 min, though platelet cyclic [3H]AMP remained above control values. In these experiments, the time course of the changes in platelet cyclic [3H]AMP correlated closely with values for blood prostacyclin obtained previously (Haslam, R.J., and M.D. McClenaghan, 1981, Nature [Lond.]., 292:364-366). Prostacyclin also had a biphasic effect on the release of [14C]serotonin when added to citrated blood in vitro, though both the increase in sensitivity to platelet-activating factor and the return of platelet cyclic [3H]AMP towards control values took place more slowly. At all times, addition of platelet-activating factor decreased platelet cyclic [3H]AMP towards but not below the control level observed in the absence of prostacyclin. Our results indicate that although transient increases in platelet cyclic AMP cause an immediate decrease in platelet responsiveness in vivo or in vitro, a period of enhanced platelet sensitivity follows as platelet cyclic AMP falls. PMID- 2991337 TI - Human alveolar macrophages produce a fibroblast-like collagenase and collagenase inhibitor. AB - Human macrophages have been implicated in connective tissue remodeling; however, little is known about their direct effects upon collagen degradation. We now report that human alveolar macrophages in culture produced both a collagenase and a collagenase inhibitor. The collagenase was secreted in latent form and could be activated by exposure to trypsin. Collagenase production could be increased three to fourfold by incubating the cells with lipopolysaccharide, but synthesis was largely unaffected by exposure to phorbol myristate acetate. By several criteria, macrophage collagenase was the same as the collagenase secreted by human skin fibroblasts: (a) they were antigenically indistinguishable in double immunodiffusion; (b) both degraded type III collagen preferentially to type I, had little activity against type II collagen, and none against types IV and V, and (c) their affinity for susceptible collagens was equivalent, Michaelis constant = 1-2 microM. Collagenase inhibitory activity was also present in the macrophage-conditioned medium, and was accounted for by an antigen that showed immunologic and functional identity with the collagenase inhibitor secreted by human skin fibroblasts. The amount of inhibitor released by unstimulated cells, approximately 100 ng/10(6) cells per 24 h, was substantially augmented by both phorbol and lipopolysaccharide, although considerable variability in response to these agents was observed between macrophage populations derived from different subjects. As negligible quantities of collagenase or collagenase inhibitor were detectable intracellularly, it appeared that both proteins were secreted rapidly after synthesis. Thus, human macrophages have the capacity to modulate collagen degradation directly by production of collagenase and collagenase inhibitor. PMID- 2991340 TI - Evidence for alteration of the vitamin D-endocrine system in obese subjects. AB - Serum immunoreactive parathyroid hormone (PTH) is increased in obese as compared with nonobese subjects and declines with weight loss. To determine whether alteration of the vitamin D-endocrine system occurs in obesity and whether ensuing secondary hyperparathyroidism is associated with a reduction in urinary calcium, a study was performed in 12 obese white individuals, five men and seven women, and 14 nonobese white subjects, eight men and six women, ranging in age from 20 to 35 yr. Body weight averaged 106 +/- 6 kg in the obese and 68 +/- 2 kg in the nonobese subjects (P less than 0.01). Each of them were hospitalized on a metabolic ward and were given a constant daily diet containing 400 mg of calcium and 900 mg of phosphorus. Whereas mean serum calcium, serum ionized calcium, and serum phosphorus were the same in the two groups, mean serum immunoreactive PTH (518 +/- 48 vs. 243 +/- 33 pg/ml, P less than 0.001), mean serum 1,25 dihydroxyvitamin D [1,25(OH)2D] (37 +/- 2 vs. 29 +/- 2, P less than 0.01), and mean serum Gla protein (33 +/- 2 vs. 24 +/- 2 ng/ml, P less than 0.02) were significantly higher, and mean serum 25-hydroxyvitamin D (25-OHD) (8 +/- 1 vs. 20 +/- 2 ng/ml, P less than 0.001) was significantly lower in the obese than in the nonobese men and women. Mean urinary phosphorus was the same in the two groups, whereas mean urinary calcium (115 +/- 10 vs. 166 +/- 13 mg/d, P less than 0.01) was significantly lower, and mean urinary cyclic AMP (3.18 +/- 0.43 vs. 1.84 +/- 0.25 nM/dl GF, P less than 0.01) and creatinine clearance (216 +/- 13 vs. 173 +/- 6 liter/d, P less than 0.01) were significantly higher in the obese than in the nonobese individuals. There was a significant positive correlation between percentage of ideal body weight and urinary cyclic AMP (r = 0.524, P less than 0.01) and between percentage of ideal body weight and serum immunoreactive PTH (r = 0.717, P less than 0.01) in the two groups. The results provide evidence that alteration of the vitamin D-endocrine system in obese subjects is characterized by secondary hyperparathyroidism which is associated with enhanced renal tubular reabsorption of calcium and increased circulating 1,25(OH)2D. The reduction of serum 25-OHD in them is attributed to feedback inhibition of hepatic synthesis of the precursor by the increased serum 1,25(OH)2D. PMID- 2991341 TI - Leukotriene C4 binds to human glomerular epithelial cells and promotes their proliferation in vitro. AB - In human and experimental glomerulonephritis, glomerular hypercellularity results both from accumulation of macrophages and proliferation of resident glomerular cells. The recent identification of macrophage-derived factors that stimulate mesangial and epithelial cell proliferation suggests that these factors might contribute to the hypercellularity. To determine the identity of such macrophage derived growth factors, we studied the effect of leukotrienes (LTs), products that are released from macrophages and leukocytes, on proliferation of human glomerular epithelial cells in culture. Dose-dependent (1-100 nM) stimulation of [3H]thymidine incorporation, an index of cell proliferation, was observed in cells incubated with the sulfidopeptide LTs, LTC4 and LTD4, but not with LTB4. The response was 248 and 172% of control values at 100 nM LTC4 and LTD4, respectively. This effect of LTC4 was abolished by FPL 55712. Subsequent binding studies demonstrated that glomerular epithelial cells possess specific receptors for LTC4. [3H]LTC4 bound rapidly at 8 degrees C to the cells. There was a plateau after 40 min incubation. Maximum specific binding was 70-90% of total binding. Specific binding was totally reversible with addition of an excess of unlabeled LTC4. Analysis of time-course association slopes at two concentrations of [3H]LTC4 and of the competition between a single concentration of [3H]LTC4 and increasing concentrations of unlabelled LTC4 allowed calculation of dissociation constants (Kd) of 220 and 217 nM, respectively. Both LTD4 and LTE4 exhibited ED50 values that were at least one order of magnitude higher than for LTC4. Thus, our findings suggest that LTC4 binds to specific receptors of glomerular epithelial cells, promotes proliferation of these cells, and could contribute to epithelial hypercellularity found in glomerulonephritis. PMID- 2991339 TI - Effect of hydrogen peroxide on prostaglandin production and cellular integrity in cultured porcine aortic endothelial cells. AB - Oxidative damage to the vascular endothelium may play an important role in the pathogenesis of atherosclerosis and aging, and may account in part for reduced vascular prostacyclin (PGI2) synthesis associated with both conditions. Using H2O2 to induce injury, we investigated the effects of oxidative damage on PGI2 synthesis in cultured endothelial cells (EC). Preincubation of EC with H2O2 produced a dose-dependent inhibition (inhibitory concentration [IC50] = 35 microM) of PGI2 formation from arachidonate. The maximum dose-related effect occurred within 1 min after exposure although appreciable H2O2 remained after 30 min (30% of original). In addition, H2O2 produced both a time- and dose-dependent injury leading to cell disruption, lactate dehydrogenase release, and 51Cr release from prelabeled cells. However, in dramatic contrast to H2O2 effects on PGI2 synthesis, loss of cellular integrity required doses in excess of 0.5 mM and incubation times in excess of 1 h. The superoxide-generating system, xanthine plus xanthine oxidase, produced a similar inhibition of PGI2 formation. Such inhibition was dependent on the generation of H2O2 but not superoxide in that catalase was completely protective whereas superoxide dismutase was not. H2O2 (50 microM) also effectively inhibited basal and ionophore A23187 (0.5 microM) stimulated PGI2 formation. However, H2O2 had no effect on phospholipase A2 activity, because ionophore A23187-induced arachidonate release was unimpaired. To determine the effects on cyclooxygenase and PGI2 synthase, prostaglandin products from cells prelabeled with [3H]arachidonate and stimulated with ionophore A23187, or products formed from exogenous arachidonate were examined. Inhibition of cyclooxygenase but not PGI2 synthase was observed. Incubation of H2O2-treated cells with prostaglandin cyclic endoperoxide indicated no inhibition of PGI2 synthase. Thus, in EC low doses of H2O2 potently inhibit cyclooxygenase after brief exposure whereas larger doses and prolonged exposure are required for classical cytolytic effects. Surprisingly, PGI2 synthase, which is known to be extremely sensitive to a variety of lipid peroxides, is not inhibited by H2O2. Lipid solubility, enzyme location within the EC membrane, or the local availability of reducing factors may explain these results, and may be important determinants of the response of EC to oxidative stress. PMID- 2991342 TI - Antigenic characterization of human hepatocellular carcinoma. Development of in vitro and in vivo immunoassays that use monoclonal antibodies. AB - Several libraries of monoclonal antibodies have been produced by immunization of Balb/c mice with single cell suspensions of nontrypsin-treated human hepatocellular carcinoma cell (HCC) lines in order to study the antigenic properties of transformed hepatocytes. The antibodies were characterized with regards to specificity for hepatoma-associated antigens and their capability for use as reagents in radioimmunoassays (RIAs) and tumor localization in vivo. Three such antibodies namely, P215457, PM4E9917, P232524 of the IgG2a, IgG2a, and IgG1 isotypes, respectively, not only recognized separate and distinct antigenic determinants on four human hepatoma cell lines but also reacted with epitopes present on chemically induced rat hepatoma cell lines. In contrast, only 1 of 38 other human malignant and transformed cell lines demonstrated reactivity with the three antibodies; normal human tissues were also found to be unreactive. Monoclonal antibody P215457 densely stained the plasma membrane by indirect immunofluorescence, showed rapid binding activity to HCC cells in suspension, and precipitated a 50,000-mol wt cell surface protein; antibody PM4E9917 also stained the plasma membrane and precipitated a 65,000-mol wt protein, whereas P232534 recognized cytoplasmic antigenic determinants. With these antibodies "simultaneous sandwich" RIAs were established that detect soluble hepatoma associated antigens in culture supernatants. Finally, the Fab fragment of P215457 was found to be useful in tumor localization in vivo. This antibody fragment when labeled with 131I was shown to localize by radionuclide-imaging studies in human hepatoma grown in nude mice. Thus, these investigations demonstrate that monoclonal antibodies may be produced against epitopes that reside almost exclusively on transformed hepatocytes and such antibodies may be successfully employed in the development of in vitro and in vivo immunoassays. PMID- 2991344 TI - An account of the management of a potential environmental/medical crisis by a local health department. AB - This paper is an account of what steps were necessary in the epidemiological and medical investigation of an alleged cyanide outbreak by a local health department. It details, in chronological sequence, the events, groups, and individuals who impacted a situation which, at various points, caused considerable anxiety in the community. PMID- 2991343 TI - Resistance of human tumor cells in vitro to oxidative cytolysis. AB - Nine human cell types, six of them malignant, displayed a marked resistance to lysis by hydrogen peroxide (LD50, 2-20 mM). Of the reactive oxygen intermediates generated extracellularly, only H2O2 lysed all the cell types. OH was lytic to one of four, OI- to one of one, and O-2 to none of four cell types tested. Resistance to oxidative lysis did not correlate with specific activity of catalase, glutathione (GSH) peroxidase, other peroxidases, or glutathione disulfide reductase, or with specific content of GSH. Resistance to H2O2 seemed to occur via mechanisms distinct from those responsible for cellular consumption of H2O2. Consumption was inhibitable by azide and was probably due to catalase in each cell type. In contrast, resistance to oxidative lysis occurred via distinct routes in different cells. One cell type used the GSH redox cycle as the primary defense against H2O2, like murine tumors previously studied. Other cells seemed to utilize catalase as the major defense against H2O2. Nonetheless, with both catalase and the GSH redox cycle inhibited, all the human cells tested exhibited an inherent resistance to oxidative lysis, that is, resistance independent of detectable degradation of H2O2. PMID- 2991345 TI - MR imaging: clinical use of the inversion recovery sequence. AB - The properties of the inversion recovery (IR) sequence are considered and its use in clinical practice is illustrated. The effect of changing repetition time, inversion time (TI), and echo time; the method of data encoding; the type of data collection; and the method of image processing are analysed. Normal appearances and clinical examples in the central nervous system and the remainder of the body are used to illustrate the many options available with this sequence. The short TI IR sequence has advantages in magnetic resonance imaging of the body, and medium TI sequences are of value in localisation in the brain and in demonstrating contrast enhancement. Long TI sequences can be used in pediatrics and for separating tumour and oedema. Suppression or partial suppression of fat and fluid signals are two useful options with IR sequences. PMID- 2991346 TI - Comparative anatomo-functional imaging of two neuroreceptors and glucose metabolism: a PET study performed in the living baboon. AB - Regional specialization of two types of neuroreceptor--opiate and dopamine--was mapped by positron emission tomography (PET) in the subcortical gray matter of the living baboon and analyzed in comparison with glucose metabolism. The three radiopharmaceuticals used--[18F]3-acetylcyclofoxy, [11C]3-N-methylspiperone, and [18F]2-fluoro-2-deoxyglucose--traced opiate- and dopamine-receptor distribution and regional glucose metabolism, respectively. A combination of high spatial resolution (6-7 mm) and the intrinsic sensitivity of the PET method made possible the detection of nanomolar concentrations of neuroreceptors and micromolar concentrations of metabolized glucose, with different and distinctive anatomo functional distributions in thalami and basal ganglia. PMID- 2991347 TI - MR imaging of the liver in primary hepatocellular carcinoma. AB - In 10 patients with hepatoma, magnetic resonance (MR) and CT of the liver were subjectively compared and correlated with surgical or autopsy findings. In five cases MR defined the extent of the tumor better. Magnetic resonance was particularly useful in differentiating the tumor from otherwise abnormal areas of the liver, mostly focal cirrhosis. Magnetic resonance has the advantage of demonstrating major vessels in relation to the hepatoma without injection of any contrast agent. Calcifications well visualized on CT are not seen on MR. The lack of dynamic bolus CT in the majority of our cases as well as our inability to examine the entire liver with all three MR pulse sequences because of time restraint are significant limitations of this retrospective study. PMID- 2991348 TI - Forskolin modulates cyclic AMP generation in the rat myometrium. Interactions with isoproterenol and prostaglandins E2 and I2. AB - In the intact rat myometrium, forskolin stimulated cyclic AMP generation and markedly potentiated increases in cyclic AMP caused by isoproterenol, prostaglandin E2 and prostacyclin. The diterpene increased the maximal responses and lowered the EC50 for both isoproterenol- and prostaglandin-stimulated cyclic AMP accumulation. Forskolin did not modify the Ki for the beta-adrenergic antagonist propranolol. Activation of cyclic AMP generation by forskolin was biphasic with respect to concentration; the major response being mediated by a low affinity interaction (Kapp 28 microM) and a minor effect being due to an interaction with a high affinity site (Kapp 0.5 microM). By contrast, the synergistic effect of the diterpene with isoproterenol, prostaglandin E2 as well as with cholera toxin, involved a single component of high affinity (Kapp 0.5 to 2 microM), which was thus considered to be associated with the activated complex of the cyclase catalytic subunit and the guanine nucleotide regulatory protein. Forskolin could further partially maintain isoproterenol-mediated synergism in a beta-adrenergic desensitized tissue. In myometrial membrane preparations, forskolin stimulated adenylate cyclase activity but failed to potentiate isoproterenol- and prostaglandin E2-mediated activation. PMID- 2991349 TI - Receptor (norepinephrine), P-site (2',5'-dideoxyadenosine), and calcium-mediated inhibition of prostaglandin and forskolin-activated cyclic AMP-generating systems in human platelets. AB - Prostaglandin D2, 2-chloroadenosine, forskolin and combinations of these agents increase cyclic AMP-levels in intact human platelets. The inhibition of activated cyclic AMP-generating systems by 1) alpha2-adrenergic receptor-mediated hormonal input (norepinephrine), 2) a P-site agent (2',5'-dideoxyadenosine) and 3) a divalent cation (calcium) were examined: 1) Norepinephrine produces non competitive inhibition of both forskolin and prostaglandin D2(PGD2)-stimulated cyclic AMP-accumulation in intact human platelets. The Ki values for norepinephrine versus forskolin, PGD2 and 2-chloroadenosine are similar in magnitude, while the Ki versus a forskolin-PGD2 combination is approximately 10 fold greater. Onset of inhibition by norepinephrine of the PGD2-response is several fold faster than for the forskolin-response. When platelets stimulated by the forskolin and PGD2 combination are exposed to norepinephrine, there is a transient increase in levels of cyclic AMP due to the potentiation of a minor beta-adrenergic component. This stimulation is followed by inhibition. 2) 2',5' Dideoxyadenosine produces a non-competitive inhibition of the forskolin-response with a Ki of 110 microM. The inhibition of the PGD2-response by 2',5' dideoxyadenosine is competitive with a Ki of 6-13 microM, while inhibition of the forskolin-PGD2 response has a Ki of 30 microM. Onset of inhibition by 2',5' dideoxyadenosine is identical for forskolin or PGD2-stimulated platelets. There is a lag period for inhibition of platelets stimulated with the forskolin-PGD2 combination. The PGD2-forskolin combination appears to stabilize the cyclic AMP generating system of platelets against inhibition by either norepinephrine or 2',5'-dideoxyadenosine. 3) Calcium ions cause a similar inhibition of cyclic AMP generation in intact platelets, regardless of the type of stimulation. PMID- 2991350 TI - Determination of cyclic AMP-dependent protein kinase subunits by an immunoassay reveals a different subcellular distribution of the enzyme in rat parotid than does determination of the enzyme activity. AB - The distribution of cyclic cAMP-dependent protein kinase in subcellular fractions of rat parotid was determined by two independent biochemical methods, measurement of kinase catalytic activity or by quantitation of the catalytic and regulatory subunits in an enzyme-linked immunosorbent assay using monospecific antibodies. The major amount (85%) of the catalytic activity was found associated with the 100,000 g soluble fraction, whereas only 1/3 of the total catalytic subunit was demonstrated in the soluble fraction by the immunoassay. The immunoassay results furthermore indicated that approximately 50% of the total cellular protein kinase was associated with the extranuclear particulate fraction and that the predominant form of the kinase in the particulate fractions was the type II isoenzyme. The reasons for the differences in the distribution of the protein kinase demonstrated by the two methods were examined. Incomplete extraction of membrane-bound protein kinase and the influence of membrane localized ATPases on activity measurements were, at least in part, responsible for the low percentage of kinase activity measured in the particulate fractions. These results emphasize that the precise quantitation of protein kinase subunits merits investigation by more than one method. For the parotid, the finding that approximately 2/3 of the total catalytic subunit may be particulate associated provides additional evidence that cyclic AMP could be involved in membrane mechanisms of hormone regulated secretion. PMID- 2991351 TI - A monoclonal antibody to a membrane component that interacts with the beta adrenergic receptor. AB - A monoclonal antibody has been obtained using a combination of in vivo and in vitro immunization with a digitonin extract of purified plasma membranes from frog erythrocytes. This antibody was found to immunoprecipitate a fraction of solubilized beta-adrenergic receptor (labeled with 125I iodohydroxybenzylpindolol) derived from frog erythrocytes and a few other sources. This immunoglobulin also significantly activated adenylate cyclase in isolated erythrocyte plasma membranes measured in the presence of GTP or GTP plus isoproterenol. Immunoprecipitation of labeled erythrocyte surface proteins and immunoblotting of the digitonin-extract of erythrocyte plasma membrane revealed that the antibody interacted with a protein with a Mr = 43,000 and pI = 6.2 32P ADP-ribosylated alpha subunit of Ns (with Mr = 44,000) solubilized from frog erythrocyte membranes failed to be immunoprecipitated by the antibody. Thus the antigenic protein is distinct from the beta-adrenergic binding site and alpha subunit of Ns and therefore may be an unidentified component of the beta adrenergic receptor-adenylate cyclase system. This monoclonal antibody may be a useful tool for future studies for topological and functional interactions between the beta-adrenergic receptor and other membrane components. PMID- 2991352 TI - Epithelial nevi and benign tumors of the skin and their associated systemic conditions. PMID- 2991353 TI - A case of granuloma annulare and sarcoidosis. PMID- 2991354 TI - Effects of a compressive nonlinearity in a cochlear model. AB - A compressive nonlinearity located at the hair cell-nerve fiber complex of a cochlear model is shown to generate realistic single-tone responses, as well as to cause lifelike "synchrony suppression." PMID- 2991355 TI - On the extraction of the signal-excitation function from a non-Poisson cochlear neural spike train. AB - In two recent papers in this Journal, Johnson and Swami [J. Acoust. Soc. Am. 74, 493-501 (1983)] and Gaumond et al. [J. Acoust. Soc. Am. 74, 1392-1398 (1983)] have given procedures for estimating the signal-excitation function s(t) conveyed by a non-Poisson cochlear neural spike train with a known neural recovery function r(t - t'). For a delayed step form of r(t - t'), we show how s(t) may be directly (noniteratively) determined from the post-stimulus-time (PST) histogram response. We also show that for more general recovery functions, s(t) may be determined from the PST histogram response by using a simple iterative method. PMID- 2991356 TI - Rhythm-related changes in pituitary-adrenal function in depression. AB - We measured plasma ACTH and cortisol at 20-min intervals for 24 h in depressed patients and healthy control subjects. The data were analyzed by the PULSAR program to quantitate the number of hormone pulses, their amplitude, length, maximum and interval. We found that in both healthy and depressed subjects the circadian pattern of pituitary-adrenal activity is the result of significant time related changes in pulse amplitude with no change in pulse frequency. Depressed patients who had an abnormal response to dexamethasone also had changes in pituitary-adrenal rhythm in the unmedicated state. These included ACTH and cortisol pulses whose amplitude, maximum and duration were greater than in the controls as well as a phase advance in the cortisol circadian rhythm. Some of those features were shared by patients who responded normally to dexamethasone suggesting that rhythm-related indices of pituitary-adrenal function may be a more sensitive index of disturbed pituitary adrenal regulation than the Dexamethasone Suppression Test. PMID- 2991357 TI - Tomographic gated blood pool radionuclide ventriculography: analysis of wall motion and left ventricular volumes in patients with coronary artery disease. AB - The use of planar radionuclide ventriculography to evaluate global and segmental ventricular function is limited by the superimposition of structures in some projections and the gross segmental resolution of the planar technique. Preliminary reports have suggested the feasibility of tomographic gated radionuclide ventriculography with rotating detector systems. This study tested the hypotheses that 1) tomographic radionuclide ventriculography detects segmental dysfunction at rest not identified with multiview planar studies and single plane contrast ventriculography, and 2) ventricular volumes and ejection fraction calculated from these studies provide data similar to those obtained with angiography and planar radionuclide ventriculography. Gated blood pool tomograms were acquired over 180 degrees at 15 frames per cardiac cycle during the initial 90% of the cardiac cycle. Compared with the multiview planar technique tomographic ventriculography showed an increased sensitivity for detecting left ventricular segments with significant coronary artery stenosis (97 versus 74%, p less than 0.025) without any loss in specificity. Compared with both planar radionuclide and contrast ventriculography, tomographic radionuclide ventriculography also detected more noninfarcted left ventricular segments supplied by stenosed coronary arteries (81 versus 39 and 32%, respectively, p less than 0.01). Tomographic radionuclide ventriculographic measurements of left ventricular volumes and ejection fraction showed close correlations with angiographic and planar radionuclide determinations. Gated blood pool tomography is a sensitive method for the evaluation of segmental wall motion and an accurate method for the measurement of global left ventricular volumes and ejection fraction. PMID- 2991358 TI - Elevated leukocyte phosphodiesterase as a basis for depressed cyclic adenosine monophosphate responses in the Basenji greyhound dog model of asthma. AB - The BG dog manifests various characteristics of human asthma, including airway hyperreactivity to low concentrations of methacholine. Studies have suggested that airway hyperreactivity in asthma is related to inadequate intracellular cAMP responses. We studied cAMP characteristics in MNL from 19 BG and 14 mongrel dogs. beta-Adrenergic receptors were assessed by 125I CYP in the presence and absence of propranolol. The responses of cAMP to ISO were measured by radioimmunoassay. Adenylate cyclase activity was determined in homogenized MNL preparations by cAMP generation. PDE activity was quantitated by radioenzyme assay. Mongrel dog leukocyte ISO-stimulated cAMP levels doubled, whereas there were negligible increases in MNL from BG dogs. Basal PDE levels were higher in BG dogs than in mongrel dogs. The PDE inhibitor Ro 20-1724 restored ISO-stimulated cAMP responses in MNL of BG dogs. Adenylate cyclase activity was not lower in MNL homogenates from BG dogs than in mongrel dogs. Cells from both BG and mongrel dogs demonstrated similar receptor numbers and affinities of saturable, specific beta adrenergic binding over a 10 pM to 400 pM range. Our results suggest that depressed cAMP responses in BG dogs are due to high PDE activity rather than to a defect in the beta-adrenergic receptor adenylate cyclase system. PMID- 2991359 TI - Adrenergic receptors: possible mechanism of inverse regulation of alpha- and beta receptors. AB - Many physiologic and pathologic conditions, including bronchial asthma, are associated with inverse changes in alpha- and beta-receptor-mediated responses in various tissues. The direction of the change elicited by a given stimulus is tissue specific, as exemplified by the actions of thyroid hormones: In the rat heart, hypothyroidism reduces beta- and increases alpha-receptor responses, whereas in the rat liver it has the opposite effects. A similar increase in beta- and decrease in alpha-receptor responses in the rat liver is triggered by a number of different conditions, including glucocorticoid deficiency, that appear to represent lower levels of cellular differentiation. Among these, incubation of isolated hepatocytes in a serum-free buffer triggers the conversion of the receptor response in vitro within 4 hours, without parallel changes in the density or affinity of receptor binding sites. This change can be acutely reversed by an endogenous inhibitor of membrane phospholipase A2, or accelerated by an activator of phospholipase A2, suggesting that changes in the activity of this enzyme are involved in the conversion of the hepatic adrenoceptor response. The glucocorticoid-induced increase in beta-receptors in cultured human lung adenocarcinoma cells also appears to be mediated indirectly through the induction of an endogenous inhibitor (lipomodulin) of membrane phospholipase A2. The possible relevance of altered membrane phospholipid metabolism in the pathomechanism of asthma and in the associated glucocorticoid-sensitive changes in adrenergic receptor mechanisms is discussed. PMID- 2991360 TI - Future pharmacologic agents and maneuvers in asthma therapy. AB - Future therapeutic avenues in the treatment of asthma include the development of drugs to prevent mediator generation or release, to prevent receptor recognition of mediators, and to block end-organ responses to receptor occupancy; maneuvers to prevent or reverse IgE production; and therapies directed at preventing secondary inflammatory leukocyte activation. These new therapies should be used only after currently available drugs, education, immunotherapy, and supportive care are maximally employed, as these latter maneuvers are in the vast majority of patients efficacious, well tolerated, and safe. PMID- 2991362 TI - Improved localization of intracellular sites of phosphatases using cerium and cell permeabilization. AB - A simple permeabilization method has been developed that allows for intracellular localization of acid phosphatase in neutrophils and several types of tissue culture cells with cerium. This permeabilization procedure also facilitates intracellular alkaline phosphatase localization in neutrophils without the loss of cell surface reaction in this cell type. Only the cell surface reaction was detected in the absence of permeabilization. Glutaraldehyde-fixed cells were permeabilized with detergent during the cytochemical reaction. Triton X-100 at 0.0001-0.0002% gave the best results for the enzymes and cell types tested. PMID- 2991361 TI - Clinico-pathologic conference: 60-year-old man with weakness and respiratory failure. PMID- 2991363 TI - Involvement of Golgi apparatus and a restructured nuclear envelope during biogenesis and transport of herpes simplex virus glycoproteins. AB - Following infection of BHK-21 cells with Herpes simplex virus type 1 (HSV-1), progeny nucleocapsids in the nucleus acquire a glycoprotein-rich envelope by budding through host-cell nuclear membranes. To investigate the nature of the glycoprotein products assembled in the virion at the nuclear envelope, infected cells were pulse-labeled with [3H]-mannose, an oligosaccharidal core sugar, or [3H]-fucose, a terminal sugar. After various chase periods, the incorporation of these sugars was monitored by electron microscope radioautography. The results show that HSV glycoproteins accumulate very rapidly in nuclear membranes, where they exist only as core-glycosylated precursors, i.e., containing [3H]-mannose but not [3H]-fucose. [3H]-fucose grains are seen mainly over Golgi membranes and over virions located in the Golgi and in other cytoplasmic vesicular structures. Our data support a model where addition of terminal sugars (e.g., fucose) to HSV 1 glycoprotein precursors can occur at the surface of newly enveloped viral particles as the virions themselves egress from the cell via the Golgi apparatus. PMID- 2991364 TI - Lowicryl K4M embedding of brain tissue for immunogold electron microscopy. AB - We present methods for embedding brain tissue in Lowicryl K4M embedding medium and localizing antigens using postembedding immunogold techniques. After perfusion fixation with 4% paraformaldehyde and 0.1% glutaraldehyde in 0.1 M cacodylate buffer, blocks of rat brain were placed in 2% aqueous uranyl acetate for 1 hour, dehydrated in 50%, 70%, and 95% ethanol, infiltrated with Lowicryl/ethanol mixtures (1:2 for 10 min, 1:1 for 15 min) and 100% Lowicryl (20 min and 25 min). Polymerization was carried out under UV light for 24-48 hours at room temperature. Several neural antigens, including three different synaptic vesicle proteins and an enzyme associated with the postsynaptic density, were localized by this technique, indicating that this procedure may have wide applicability. PMID- 2991365 TI - Bluetongue virus and epizootic haemorrhagic disease of deer virus serotypes in northern Colombian cattle. AB - There is recent evidence of bluetongue (BT) and epizootic haemorrhagic disease (EHD) virus infection of cattle in the American tropics, including BT group reactive antibody in Colombian cattle. These observations prompted a study to determine serologically the specific BT and EHD virus types present, and time of infection and to collect Culicoides spp. as potential vectors. A prospective study of BT and EHD virus infection was done on two farms in the Colombian department of Antioquia. Sequential sampling of young cattle indicated acquisition of neutralizing antibody to BT virus serotypes 12, 14 and 17, and EHD virus serotypes 1 and 2. Insect captures showed a high association of Culicoides insignis with infected cattle. PMID- 2991367 TI - Transplacental antibodies. Part III: Maternal antibodies against polioviruses, M. parotitidis and M. parainfluenzae. AB - In 1981 we examined 247 sera for the presence of antibodies against all three types of poliovirus and 253 sera for antibodies against M. parotitidis and three types of M. parainfluenzae viruses. The sera were obtained from the cord blood of mothers between 15 and 34 years of age. All mothers were divided into four age groups, each with primipara and multipara subgroups. The rate of seropositivity for type 1 and type 2 poliovirus-specific antibody was in all age groups higher than 90%, the overall seropositivity rate for type 3 poliovirus antibody was 83.6%, with 73.1% as the lowest rate for age group of youngest mothers. Significant seropositivity variations between the primipara and multipara subgroups were recorded only for type 3 antibody in the two age groups of oldest mothers (25-29 and 30-34 years). This is consistent with the assumed booster effect of Sabin vaccine strains on mothers of families with more than one child. Antibodies specific to mumps virus were present in the cord blood of 78.6% of all mothers and the rates of seropositivity were found to rise with the increasing age. Seropositivity for M. parainfluenzae type 1-specific antibody was demonstrated in 95.8%, for type 2-specific antibody in 98.9% and for type 3 specific antibody in 100% of mothers, which is suggestive of high herd immunity levels in the population. PMID- 2991368 TI - Excretion of live attenuated polioviruses in the faeces of orally vaccinated children. Comparison of two immunization schedules. AB - The excretion of live, attenuated poliovirus vaccine strains was determined in the feces of Prague Infants home children given 10(5) PFU of type 1 and 2 and 2.10(5) PFU of type 3 vaccine in a routine annual mass campaign. The first two faeces specimens examined in each vaccinee prior to immunization were negative for the virus. A total of 476 stool specimens were collected from 37 children at weekly intervals for a period of 18 weeks. The presence of type 1 poliovirus in the faeces of children given monovalent type 1 vaccine was detectable for 9 weeks, with a maximum in first week, and the virus was isolated in 74.2% of vaccinees. The timing of bivalent type 2 and type 3 vaccine was 9 weeks after monovalent type 1 immunization. The excretion of these two types of poliovirus was found to persist for at least 6 weeks. Type 2 poliovirus was isolated in all vaccinees, type 3 in 70.4% of children. The highest percentage of children excreting type 2 poliovirus was recorded in the first week, the excretion of type 3 peaked three weeks after bivaccine administration. The excretion peaks were reached relatively early postvaccination, with type poliovirus reaching the highest titre per 1 g of faeces. After revaccination (one year later) with monovalent type 1 vaccine, the vaccine strain of type 1 poliovirus could be detected for 6 weeks and was present in the highest percentage of positive stool samples.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991369 TI - Noradrenaline release and sympathetic nervous system activity. AB - Measurements of the plasma concentration of noradrenaline, or more specifically the rate at which noradrenaline enters plasma, provide a useful guide to sympathetic nervous system function in humans. The overall rate of release of noradrenaline to plasma gives an overview of sympathetic nervous system activity (integrated nerve firing rate), detecting generalized changes, whether occurring as a reflex response, produced by drugs, or accompanying disease processes. The pattern of sympathetic nervous activation, however, is not delineated, only the net change in neurotransmitter release. Measurement of regional rates of noradrenaline release allows the clinical assessment of organ-specific sympathetic nervous tone, and consequently more penetrating analysis of sympathetic nervous system pathophysiology in disease states. The major problem in interpreting regional noradrenaline spillover measurements lies in the difficulty in differentiating those changes in noradrenaline spillover due to altered nerve firing, from those due to extraneous factors which might also affect spillover, such as the possible influence of blood flow on noradrenaline washout. PMID- 2991370 TI - 125I-Angiotensin II binding to human blood cells. AB - The binding of 125I-angiotensin II to human blood cells was investigated. Blood was drawn from healthy volunteers and platelets prepared with minimal contamination of red cells and white blood cells (less than 0.1%). Using thin layer chromatography, degradation of 125I-angiotensin II by platelets could be demonstrated in the presence of various enzyme inhibitors. However, when incubated with 1 mM diisopropylfluorophosphate (DFP) or 1 mg/ml bacitracin, no breakdown of 125I-angiotensin II could be detected. The amount of specifically bound 125I-angiotensin II increased linearly with the number of cells per tube. Binding reached a plateau within 90-120 min at 37 degrees C, and was stable thereafter. Specific binding was reversible. No binding could be detected at 4 degrees C. Specific binding of 125I-angiotensin II was saturable. Scatchard analysis of binding by platelets of healthy volunteers revealed one class of binding sites with an apparent Kd of 127 +/- 16 pM and a maximal binding capacity of 7.9 +/- 1.5 binding sites per cell. Competitive displacement of 125I angiotensin II binding by angiotensin II-analogues showed a rank order of effectiveness. Unrelated peptides, e.g. bradykinin, vasopressin and enkephalin, did not displace specifically bound angiotensin II. Human mononuclear leucocytes were prepared by a Ficoll-isopaque gradient. However, these cells could not be used for studies of specific binding, since enzymatic degradation of 125I angiotensin II could not be prevented despite addition of various enzyme inhibitors. Time-dependent uptake of 125I-angiotensin II showed no stable plateau. Thus our study shows specific binding of 125I-angiotensin II to human platelets.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991366 TI - Effect of specific humoral immunity and some non-specific factors on resistance of volunteers to respiratory coronavirus infection. AB - Thirty-three volunteers were inoculated intranasally with coronavirus 229 E, and their responses monitored by antibody rises, symptomatology and virus excretion. These were related to their pre-trial immune status as indicated by concentrations of specific antibodies and non-specific proteins in serum and nasal washings. Both circulating and local specific antibodies were associated with protection from infection and disease, but only specific IgA antibodies of either type appeared to shorten the period of virus shedding. Although total secretory IgA was significantly associated only with reduction of symptoms, total protein in nasal washings appeared to protect against infection also, indicating that other locally produced proteins, not identified, may be associated with resistance. Two of the many factors which may affect the concentration of circulating and local protective proteins and thus influence the outcome of virus inoculation, namely, sex of the volunteer and the interval since the previous cold, were examined. Male volunteers or volunteers who had had evidence of a recent respiratory infection were less likely to be infected, but if they were infected, they had lower clinical scores and stopped shedding virus earlier than the rest. These groups possessed higher concentrations of specific antibodies and non-specific proteins in their pre-challenge sera and/or nasal washings. The significance of these findings is discussed. PMID- 2991371 TI - Regulation of platelet receptors for angiotensin II in man. AB - The effects of changes in dietary intake of sodium and potassium on 125I angiotensin II binding to platelets were studied in normal subjects. We also defined binding to platelets from patients with essential hypertension and subjects with normal blood pressure. Restriction of sodium intake in normal subjects resulted in a decrease in the number of receptor sites from 6.2 +/- 0.3 sites/cell to 4.1 +/- 0.4 sites/cell (P less than 0.01) but there were no changes in affinity as measured by the Kd. Over a range of sodium intakes from 15 to 200 mmol/day there was a negative correlation between plasma concentration of angiotensin II and receptor site concentration (rs = 0.57, P less than 0.01). Changes in dietary potassium did not affect angiotensin II binding. Angiotensin II binding was also measured in 10 patients with essential hypertension (mean blood pressure [BP] 178/107 mmHg, plasma concentrations of renin [PRC] 12 +/- 2 microU/ml and angiotensin [pANG] II 14 +/- 2 pg/ml) and 10 subjects with normal blood pressure (mean BP 112/74 mmHg, PRC 13 +/- 2 microU/ml, pANG II 13 +/- 2 pg/ml). In the hypertensive patients, binding capacity and affinity (Kd = 5.0 +/- 0.6 X 10(-10) M, 5.7 +/- 0.8 sites/cell) were similar to those in the normotensive subjects (Kd = 4.9 +/- 0.8 X 10(-10) M, 5.4 +/- 0.5 sites/cell). Changes in sensitivity to angiotensin II in essential hypertension may not be determined at receptor level. Angiotensin II receptors in platelets respond to changes in sodium intake like receptors in arterial muscle. PMID- 2991372 TI - Effects of chronic peripheral sympathectomy on plasma levels of, and the pressor response to, vasopressin. AB - The purpose of the present study was to assess the effect of chronic peripheral sympathectomy in rats on plasma vasopressin (basal and dehydrated) and on pressor sensitivity to vasopressin. Sympathectomy was produced in male Sprague-Dawley rats by daily injection of guanethidine (45 mg/kg) for 9 days. Control rats received saline over the same period. Plasma vasopressin was determined by radio immunoassay, and pressor sensitivity was determined by monitoring mean arterial pressure response to graded injections of vasopressin (0.1-20 mU) in conscious rats. Sympathectomized rats showed ptosis and supersensitivity to norepinephrine, and had significantly greater basal and dehydrated plasma vasopressin levels than controls (10.3 +/- 1.5 versus 6.2 +/- 0.7, and 12.4 +/- 0.9 versus 8.6 +/- 1.0 pg/ml, s.e.m, respectively, P less than 0.05 for both). Sympathectomized rats also had an increased pressor sensitivity to vasopressin (dose response curve shifted to left, lower threshold, greater slope, P less than 0.001). Injection of a vasopressin pressor-antagonist, d (CH2)5 Me Tyr AVP, had no effect on blood pressure in control rats but caused a significant decrease of blood pressure in sympathectomized rats (15 +/- 1.0 mmHg, P less than 0.001). These results suggest that chronic peripheral sympathectomy in rats is associated with increased basal and dehydrated plasma vasopressin, and increased pressor sensitivity to vasopressin. The effect of the vasopressin antagonist suggests that vasopressin may play a role in blood pressure maintenance in sympathectomized rats. PMID- 2991373 TI - Reduced ecto-5'-nucleotidase activity and enhanced OKT10 and HLA-DR expression on CD8 (T suppressor/cytotoxic) lymphocytes in the acquired immune deficiency syndrome: evidence of CD8 cell immaturity. AB - Markedly reduced ecto-5'-nucleotidase activity was found in peripheral blood lymphocytes from 27 out of 30 homosexual men with the acquired immune deficiency syndrome (AIDS) in association with Kaposi's sarcoma (AIDS-KS; 2.67 +/- 1.70 U/10(6) cells; n = 13), opportunistic infections (AIDS-OI; 9.29 +/- 7.32; n = 7), or the AIDS-related complex (ARC; 9.82 +/- 6.12; n = 10). These values were significantly different from healthy controls (22.70 +/- 4.58; p less than 0.001). In AIDS-KS patients, both T cells and non-T cells exhibited significantly reduced ecto-5'-NT activity (p less than 0.001). AIDS-KS CD8 cells contained 20% of the mean ecto-5'-NT activity (7.04 +/- 3.53) displayed by control CD8 cells (34.07 +/- 4.86; p less than 0.001). No significant difference in enzyme level was observed between control and AIDS-KS CD4 cells (11.93 +/- 4.98 vs 7.98 +/- 3.28, respectively). In AIDS patients, lymphocyte ecto-5'-NT activity was inversely related (r = -0.518; p less than 0.01) to the absolute number of OKT10+ cells, but no correlation was found with the number of HLA-DR+ cells (r =-0.224). Two-color analysis of lymphocytes from AIDS-KS patients revealed that 75 +/- 12% of circulating CD8 cells expressed the OKT10 antigen, whereas only 10 +/- 6% of control CD8 cells did. HLA-DR antigens, which are not normally found on circulating resting T cells, were expressed in AIDS-KS CD8 cells, although to a lesser extent than OKT10. These data demonstrate that most AIDS CD8 cells differ from control CD8 cells. Although it has been suggested that these cells are activated cytotoxic or suppressor cells, the data presented here support the hypothesis they are immature. Reduced T cell ecto-5'-NT activity and enhanced expression of OKT10 and HLA-DR antigens on circulating CD8 cells, in conjunction with lack of transferrin receptor-(OKT9) and IL 2 receptor-(Tac) bearing lymphocytes, sustain this latter hypothesis. The correlation of the numerical reduction of CD4 cells with the reduced levels of ecto-5'-NT (r = 0.606; p less than 0.01) suggests that the abnormal maturation of CD8 cells seen in AIDS might be a consequence of the CD4 deficiency characteristic of this syndrome. PMID- 2991374 TI - Antiproliferative monoclonal antibodies: detection and initial characterization. AB - Two monoclonal antibodies (MAB) are described which inhibit in vitro cellular proliferation in the absence of complement or effector cells. These MAB were produced by hybridomas made from mice immunized against human B lymphoma cells. The MAB were detected by using a colorimetric assay that quantifies proliferation based on the conversion of a yellow tetrazolium salt to a purple formazan product, a reaction that occurs only in metabolically active cells with intact mitochondrial enzymes. A human B lymphoblastoid cell was used as the screening target. RBC4 is an IgM MAB that modulates and immunoprecipitates the transferrin receptor. RBG5 is an IgG1 that binds to a nonmodulating cell surface determinant different from the transferrin receptor. Both MAB are active at low concentrations (RBC4, 0.5 microgram/ml and RBG5, 0.01 microgram/ml). Immunofluorescence staining of cell lines by RBC4 and RBG5 shows little correlation with inhibition by the antibodies. They differentially inhibit the proliferation of a panel of T, B, and myeloid cell lines. Both antibodies inhibit the proliferation of alloantigen or mitogen-activated human peripheral blood lymphocytes (PBL). Unstimulated PBL are not affected by either MAB. The RB MAB each cause different morphologic changes of target cells. Whereas RBC4-inhibited cells exhibit nonspecific changes, RBG5 causes a progressive increase in the size and nuclear number of a subset of inhibited cells. PMID- 2991375 TI - Effects of various inhibitors of arachidonic acid oxygenation on macrophage superoxide release and tumoricidal activity. AB - Macrophages release a variety of arachidonic acid metabolites after treatment with various membrane triggers or particulate stimuli. We examined the role of phospholipase and lipoxygenase inhibitors in the modulation of superoxide production and tumor cytolysis by murine macrophages. Superoxide was induced by the soluble stimulus, phorbol myristate acetate (PMA), and the particulate stimulus, opsonized zymosan, and was measured by the reduction of ferricytochrome c with the use of a micro ELISA reader. Macrophage-mediated tumor cytolysis was induced by hybridoma-derived, macrophage-activating factor (MAF) and was quantitated by 51Cr release from P815 target cells. In both assays, 72-hr peptone elicited macrophages were used. Dexamethasone, and to a lesser degree hydrocortisone, inhibited superoxide release and MAF-induced tumor cytolysis. Inhibition in the superoxide assay required pretreatment with corticosteroid. Only the gold compound, auranofin, inhibited superoxide when given simultaneously with stimulant. Other phospholipase inhibitors, including mepacrine and 4 bromophenacyl bromide, and several lipoxygenase inhibitors, including BW755c, nordihydroguaiaretic acid (NDGA), and 5,8,11,14-eicosatetraynoic acid (ETYA), failed to modulate either macrophage response at nontoxic concentrations. At the concentrations tested in the tumoricidal and superoxide assays, mepacrine and 4 bromophenacyl bromide inhibited the release of 14C-arachidonic acid from macrophages stimulated with opsonized zymosan. Our data strongly suggest that corticosteroids suppress macrophage superoxide production and tumoricidal function by a nonphospholipase-dependent mechanism. PMID- 2991376 TI - Macrophage colony-stimulating factor (M-CSF) enhances the capacity of murine macrophages to secrete oxygen reduction products. AB - The capacity of macrophage colony-stimulating factor (M-CSF) to enhance respiratory burst activity in peritoneal macrophages was measured. Macrophages incubated for 48 hr or more with concentrated L cell-conditioned medium as a source of M-CSF released two to three times as much O2- in response to PMA as did unexposed macrophages. Stimulation was noted at concentrations of colony stimulating activity from 0.1 to 2000 U/ml and was maximal at 10 to 100 U/ml. Purified, endotoxin-free CSF enhanced secretion to a similar degree as unpurified L cell-conditioned medium. Release of O2- by M-CSF macrophages occurred over 60 min and was triggered by opsonized zymosan as well as PMA. H2O2 release was also enhanced in macrophages exposed to both unpurified and purified M-CSF. These data indicate that M-CSF enhances the capacity of mature macrophages to release oxygen reduction products, and they are consistent with reports that CSF can stimulate the release of other secretory products. PMID- 2991378 TI - cAMP is an essential signal in the induction of antibody production by B cells but inhibits helper function of T cells. AB - Dibutyryl cAMP and IL 1 were found to stimulate antigen-specific and polyclonal antibody production when added together to cultures of highly purified B cells. We propose that IL 1 and an elevation in cytoplasmic cAMP represent minimal signal requirements for B cell activation. In contrast to its effect on B cells, dibutyryl cAMP inhibited helper T cell activity. Cyclic AMP suppressed the production of IL 2 and T cell replacing factor (TRF) by T cells and thus abrogated the ability of helper T cells to enhance SRBC-specific antibody production by B cells. Cyclic AMP did not inhibit the generation by T cells of B cell growth factor (BCGF). BCGF, not normally detected in Con A supernatant, was found in the culture supernatant of spleen cells that were stimulated with Con A in the presence of cAMP. Our findings indicate that cAMP blocks the production of an inhibitor of BCGF activity. cAMP had no effect on the production by macrophages of IL 1. PMID- 2991377 TI - Mechanism of priming of human neutrophils by a soluble lymphoblastoid cell factor. AB - Treatment of human neutrophils (PMN) with a cytokine-like factor in the supernatants of human lymphoblastoid cells (Raji) increased the random mobility and enhanced the migration of treated cells in response to other chemoattractants nearly 21/2-fold, although the supernatant itself was not a chemoattractant. Supernatant treatment also increased the adherence of bacteria threefold and the bacterial killing fourfold compared with PMN treated with control media. In examining the metabolic basis for the enhanced bactericidal ability, we observed a significant increase in spontaneous hexose monophosphate shunt activity of Raji cell supernatant (RS)-treated neutrophils even in the absence of additional stimuli. RS-treated PMN also had significantly enhanced production of superoxide anion and chemiluminescence response upon subsequent stimulation with a variety of soluble and particulate stimuli. Unlike other agents that prime neutrophil activation, however, the factor(s) in RS did not cause degranulation. It also differed in its ability to progressively enhance PMN functions with a longer period of preincubation (up to 3 hr). These data suggest that the RS factor(s) primes neutrophils by a unique mechanism. The neutrophil-enhancing activities of RS, which are the opposite of those activities described for leukocyte inhibitory factor, eluted off a Sephacryl S-200 column at approximately 30,000 m.w. This factor expands the relationship between neutrophils and lymphocytes, and may be a useful agent to provide valuable insights into the mechanism of respiratory burst activation and regulation. PMID- 2991379 TI - Functional integrity of cytokineplasts: specific chemotactic and capping responses. AB - Cytokineplasts (CKP) are motile, membrane-bound, anucleate, granule-poor cytoplasmic fragments that are induced from human blood polymorphonuclear leukocytes (PMN) by the brief application of heat. We examined CKP with respect to specific chemotactic and capping responses, the presence of the N-formyl peptide chemotactide receptor, and evidence of respiratory burst activity and compared them with CB-cytoplasts, which are fragments created by the centrifugation of cytochalasin B (CB)-treated PMN at high speeds. Under agarose, CKP responded chemotactically to both N-formyl-methionyl-leucyl-phenylalanine (fmlp) and zymosan-activated serum; CB-cytoplasts responded to neither chemoattractant. Despite the functional differences, both fragments retained N formyl-peptide receptors as measured by affinity labeling with N-formyl-norleu leu-phe-norleu-125I-tyr-lys and autoradiography of dried SDS-PAGE gels. For studies of capping we used a murine monoclonal antibody, PMN7C3, which binds a specific, widely distributed membrane component of intact PMN, and on warming, promptly induces capping of ligand-receptor complexes. Rhodamine-conjugated PMN7C3 at 4 degrees C labeled the surface of CKP homogeneously. As the CKP warmed to 37 degrees C, label became concentrated in small fluorescent caps at the rear of migrating fragments. Although CB-cytoplasts also bound the fluorochromed antibody homogeneously in the cold, on warming they were unable to concentrate the label normally. With respect to respiratory burst activity, the situation in the two fragments was reversed: CKP did not generate superoxide anion when stimulated either with phorbol myristate acetate or with fmlp after pretreatment with CB; CB-cytoplasts, as noted earlier by other investigators, did. These two types of cytoplasts with markedly different capabilities have complementary roles in the analysis of PMN function. PMID- 2991380 TI - Demyelination induced by Theiler's virus: influence of the H-2 haplotype. AB - Demyelination induced by Theiler's virus was examined in nonrecombinant H-2 congeneic strains of mice on common backgrounds expressing independent haplotypes. Light and electron microscopy of spinal cord sections from mice with s, f, p, r, v, or q haplotypes on a C57BL/10 background showed perivascular inflammation and multifocal areas of demyelination in the white matter. The demyelination in these mice was usually associated with clinical neurologic deficits. In contrast, mice with identical genetic backgrounds but b, k, or d haplotypes showed no pathologic or clinical abnormalities. Qa or Tla haplotypes did not appear to influence demyelination. The observations support the hypothesis that susceptibility to Theiler's murine encephalomyelitis virus induced demyelination is influenced by genes that are linked to the H-2 complex. The findings suggest that the demyelination may be mediated by immune cells rather than being a direct cytolytic effect of virus on oligodendrocytes. PMID- 2991381 TI - HLA-DR beta genes vary in number between different DR specificities, whereas the number of DQ beta genes is constant. AB - Probes isolated from DR and DQ beta cDNA and genomic clones were used in hybridizations to restriction enzyme-digested DNA from human homozygous typing cells (HTC) as well as other DR homozygous cells in order to estimate the number of beta genes in the DR/DQ class II region. Varying numbers of DR beta genes were found in HTC of different DR specificities, from possibly one in DR 8 cells to three in cells of DR 2 to 7. The DR beta genes of different specificities seem to be related to one another in a distinct fashion. In contrast, all HTC contain two DQ beta genes per chromosome. The restriction site polymorphism of DQ beta genes is considerably more extensive than that of DQ serology, although one of the genes seems to be nonpolymorphic. In addition to the two DP beta genes identified previously, a minimum of three to five DQ and DR beta genes exist in the human haploid genome. PMID- 2991382 TI - Isolation and characterization of the cDNA clone and genomic clones of a new HLA class II antigen heavy chain, DO alpha. AB - From a human cDNA library constructed from a consanguineous HLA-homozygous cell line, AKIBA (HLA-A24, Bw52, DR2, Dw12, DQw1, and Cp63) (Cp63, a new SB type), a cDNA clone encoding a new HLA class II antigen heavy chain named DQ alpha was isolated, and was analyzed by Southern blot hybridization and by nucleotide sequence determination. The nucleotide sequence of the DO alpha cDNA was distinct from those of the DR alpha, the DQ alpha, and the DP alpha cDNA, but showed some characteristic features of the class II antigen alpha-chains. We also isolated and identified genomic clones specifying the DO alpha gene. Genomic analyses of cell lines with different HLA-DR serotypes with the use of the DO alpha cDNA as a probe indicated the existence of a single DO alpha gene that exhibited little restriction enzyme polymorphism. PMID- 2991384 TI - Evidence for the involvement of cytolytic macrophages in rejection of SV40 induced tumors. AB - The potential role of cytolytic macrophages in in vivo resistance to tumors induced by simian virus 40 (SV40) was evaluated in two experimental systems. First, a cell line produced by sequential in vivo passage of SV40-transformed fibroblasts through syngeneic C3H/HeJ mice was found to develop both increased neoplastic character and resistance to macrophage-mediated lysis, suggesting in vivo selection pressure against the macrophage-sensitive phenotype. In the second approach, SV40-transformed cells from C3H.OL mice, a strain that fails to produce SV40-specific cytolytic T lymphocytes (CTL), were cloned, and the cloned cells were tested for susceptibility to macrophage cytolysis in vitro. Two clones SV COL-E8 and SV-COL-F5, which represent the extremes of macrophage susceptibility and resistance, respectively, were tested for progressive growth in syngeneic C3H.OL recipients. Progression in vivo was found to correlate with resistance to macrophage cytolysis in vitro. Other in vitro measures of the neoplastic phenotype, cell division rate and anchorage-independent growth, did not predict the relative abilities of clones E8 and F5 to form tumors. Likewise, the cells were indistinguishable in their sensitivity to cytolysis by allogeneic CTL and by natural killer cells. Finally, the presence of activated macrophages in the peritoneum of mice rejecting a challenge of syngeneic SV40-transformed cells was confirmed in both CTL responder and nonresponder strains. These studies suggest that cytolytic macrophages are indeed generated during rejection of SV40-induced mouse tumors and that, in the absence of an effective anti-SV40 CTL response, resistance of the transformed cell to macrophage-mediated cytolysis can be a determining factor in in vivo tumor growth. PMID- 2991383 TI - Structure and expression of two porcine genomic clones encoding class I MHC antigens. AB - Two nonallelic porcine class I MHC (SLA) genes have been isolated and characterized. Both genes are expressed in mouse L cells, directing the synthesis of class I SLA molecules that carry common monomorphic determinants but are serologically distinct. The corresponding DNA sequences have been determined. The organization of both of these genes is similar to that of other class I genes: a leader exon, three exons encoding extracellular domains, a transmembrane exon, and three intracytoplasmic exons. The two genes are highly homologous in both exon and intron segments, with average homologies of 88% and 80%, respectively. Nucleotide changes in exon 2 are clustered, whereas those in the other exons are dispersed throughout. Comparison of the swine DNA sequences with class I genes from other species reveals a generally high conservation of exons 2, 3, 4, and 6 with lower homology in the remaining protein-encoding domains. Introns are markedly less well conserved, although moderate homology is found between swine and human class I MHC genes in both introns and 3' flanking regions. Taken together with comparisons of the deduced protein sequences, these data indicate an order of swine greater than human greater than rabbit greater than mouse in the relationship of class I genes. PMID- 2991385 TI - Simple solid-phase radioimmunoassay for human leukemia-associated cell membrane antigens. AB - In the present study, a simple solid-phase radioimmunoassay was developed to determine detergent-extracted human leukemia-associated cell membrane antigens. In the assay, 96-well microtiter plates are coated with human leukemia cell membrane antigens containing a T cell leukemia or a non-T cell leukemia antigen in the presence of a detergent, and treated with 1.6% bovine serum albumin solution. The coated antigens were reacted with an appropriate murine monoclonal antibody (mAb), i.e., SN2 or SN3, which defines a T leukemia antigen or a non-T leukemia antigen, respectively. The bound mAb is determined by a second reaction with 125I-labeled F(ab')2 of goat anti-mouse Ig. The effect of the following detergents on the assay was investigated: Nonidet P-40 (NP-40), Renex 30, deoxycholate, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and taurocholate. The best antigen dose-dependent antibody binding results were obtained using the plates coated with antigens in the presence of taurocholate, a rarely used detergent, whereas no significant antibody binding was observed in the assays using 2 nonionic detergents, i.e., NP-40 and Renex 30. In addition, the usefulness of the present assay with taurocholate during the purification of the antigens was demonstrated. It is very likely that the present solid-phase radioimmunoassay using taurocholate will also be useful in determining other non leukemia cell membrane antigens. PMID- 2991386 TI - Use of a self-generating percoll gradient and single cell cytotoxicity assay to identify tumor-lytic properties of inflammatory neutrophils. AB - Within a murine model of regional immunotherapy, the cytolytic potential of peritoneal neutrophils could not be confirmed or quantified using routine techniques of cell separation and chromium release assays. We, therefore, developed procedures for the enrichment of neutrophils and estimation of the frequency of killer cells. Peritoneal exudate cells from mice injected with Corynebacterium parvum were fractionated on a self-generating Percoll gradient to enrich for neutrophils and deplete macrophages. A significant enrichment of neutrophils (greater than 90%) was obtained in a band corresponding to a density of 1.088 with a recovery of 35-50% of input. Neutrophil-enriched cell populations were then mixed with tumor cells to examine neutrophil-target interactions at the single cell level. Conjugates of neutrophils and tumor targets were obtained and the majority were lytic. With the aid of trypan blue staining and safranin counterstaining, it was possible to distinguish effector cells from targets and neutrophils from other host cells. The frequency of conjugates was dependent upon the effector to target cell ratio and was not affected by changes in temperature (range 4-30 degrees C). The post-binding lytic events were initiated rapidly after conjugation and tumor lysis was completed within 30 min. The lytic events occurred optimally between 25 degrees and 37 degrees C. The present studies support the role of neutrophils in tumor lysis following administration of an immunoadjuvant. The techniques described are important to further study the role of neutrophils in disease states as well as the underlying mechanisms of neutrophil-mediated tumor cytotoxicity. PMID- 2991387 TI - Development of enzyme-linked immunosorbent assays for measurement of leukotrienes and prostaglandins. AB - Enzyme-linked immunosorbent assays (ELISAs) were developed for the leukotrienes LTC4 and LTB4 and the prostaglandins 6-keto PGF1 alpha and thromboxane (TxB2). In an indirect assay procedure for all 4 eicosanoids a BSA conjugate of the leukotrienes or an ovalbumin conjugate of the prostaglandins was absorbed to polystyrene microtiter plates. Samples containing the respective eicosanoids were incubated in the coated wells with specific rabbit antisera. The wells were then incubated successively with a goat anti-rabbit antibody linked to fluorescein and a rabbit anti-fluorescein antibody linked to alkaline phosphatase. The resultant assays for LTC4, LTB4, 6-keto PGF1 alpha, and TxB2, gave steep, sensitive inhibition curves; IC50s were 0.2, 10, 1, and 0.4 pmol respectively with minimal cross-reactivity to other eicosanoids. The sensitivities and specificities were comparable to those found in the RIA, and the levels determined in this assay correlate well with those determined in non-immunological assays. PMID- 2991388 TI - Instability of succinyl ester linkages in O2'-monosuccinyl cyclic AMP-protein conjugates at neutral pH. AB - Chromatographic and immunological evidence is presented regarding the hydrolysis of the ester linkage of O2'-monosuccinyl cyclic AMP in neutral solutions. Such hydrolysis occurs whether the nucleotide derivative is present in free form in solution or conjugated through its succinyl carboxyl group via an amide bond to proteins. The latter process apparently occurs when succinyl cyclic AMP is conjugated to human serum albumin for use as an immunogen in the production of anti-cyclic AMP antibodies and when the derivative is coupled to the enzyme glucose-6-phosphate dehydrogenase (E.C. 1.1.1.49). The enzyme conjugate has been used in developing a homogeneous enzyme immunoassay for cyclic AMP. Inhibition of the catalytic activity of enzyme-cyclic AMP conjugates by anti-cyclic AMP antibody decreases with time, apparently due to the loss of cyclic AMP from enzyme-cyclic AMP conjugates stored in neutral solutions. In addition, the ability of free cyclic AMP to completely reverse the inhibition process decreases with time because of the presence of antibodies in the anti-cyclic AMP sera that apparently inhibit enzyme activity because of their binding specificity for the residual succinate-protein determinant sites of the enzyme conjugates. Lyophilization of the conjugates immediately after preparation helps to overcome the problem; however, in vivo hydrolysis of immunogens prepared with the succinyl cyclic AMP derivative may always occur. The consequence of this hydrolysis reaction and the subsequent formation of anti-succinyl-protein antibodies will be discussed with regard to existing RIAs for cyclic AMP and a new homogeneous enzyme immunoassay for the nucleotide. PMID- 2991389 TI - [Balkan nephropathy and transitional cell carcinoma. Pathogenetic problems and the early phase of cancerogenesis]. AB - Balkan Nephropathy belongs to the group of primary chronic interstitial nephropathy with tubular functional abnormalities at early clinical stage. This disease may occur at any age and may affect all members of the same family including the children existing in them in latent form. The coincidental occurrence of Balkan Nephropathy and urothelial carcinoma has been very often observed. The actual problems of early Balkan Nephropathy pathogenesis and carcinogenesis are considered. PMID- 2991390 TI - Papillomavirus infection of the anogenital region: correlation between histology, clinical picture, and virus type. Proposal of a new nomenclature. AB - The clinical and histologic picture of 84 anogenital condylomatous and condyloma like lesions of both sexes were analyzed in an effort to establish a correlation to the different papillomavirus (PV) types. The presence of human papillomavirus (HPV)-specific DNA sequences was confirmed through molecular hybridization and the presence of PV structure antigens was verified in thin sections by means of a group-specific anti-PV-antiserum using the peroxidase-antiperoxidase (PAP) technique. Three distinct clinical forms harboring distinct HPV types were distinguished: (1) Condylomata acuminata in which HPV-6 DNA was present in 37 of 59 samples and HPV-11 DNA in only 13 of 59 samples. HPV-16 DNA was not detected at all and 9 condylomatous lesions remained unclassified. (2) Flat condyloma-like lesions, where HPV-6 and HPV-11 were associated with lesions of low epidermal atypia in 8 and in 2 of 18 cases, respectively, and where HPV-16 was associated exclusively with 6 of 18 such lesions with severe atypia, called bowenoid papulosis. (3) Pigmented papules where HPV-16 was detected twice in lesions of bowenoid papulosis and HPV-11 in 2 of the benign pigmented lesions. The fourth clinical manifestation of genital papillomavirus infections--the so-called condylomata plana--was not available for virologic analysis. Histologically 5 different koilocytotic features were determined which could not be correlated either with one of the clinical pictures or with a specific PV type. HPV-16, however, was found frequently in non-koilocytotic lesions exhibiting the features of severe epithelial atypia known in bowenoid papulosis. The existence of PV structure antigens in these lesions could not be verified using the indirect immunoperoxidase--PAP-technique--in contrast to the koilocytotic lesions where clear evidence of the presence of HPV was proved in 36 of 56 (64.3%) of the cases. PMID- 2991391 TI - Stimulation of skin fibroblast collagenase production by a cytokine derived from basal cell carcinomas. AB - Our previous studies of human basal cell carcinomas (BCC) revealed increased skin collagenase in vivo. Immunocytochemically the collagenase was localized to adjacent stroma, not to the tumor cells. When grown in culture, skin fibroblasts derived from tumor stroma showed a 3- to 4-fold increase in collagenase for the first 10-14 mean population doublings, after which collagenase expression reverted to control levels. These studies suggested that tumors stimulated adjacent fibroblasts to produce more collagenase. In the present study we sought direct evidence for epithelial-stromal interaction in this neoplasm. Under dissecting microscopy tumor islands were freed of stroma, homogenized, sonicated, and centrifuged to remove insoluble tissue. Tumor extracts were incubated with monolayer cultures of normal human skin fibroblasts to assess their effect on collagenase synthesis in these target cells. Culturing the fibroblasts for 24 h in the presence of individual BCC extracts resulted in a 1.6- to 3-fold increase in trypsin-activatable collagenase in the culture medium. This was paralleled by an equal increase in immunoreactive protein, suggesting enhanced enzyme synthesis. There was no change in the activity per immunoreactive protein, indicating a catalytically unaltered enzyme. Gel filtration of pooled BCC extracts showed that the stimulatory activity was contained in eluent fractions of Mr approximately 19Kd. These data suggest that BCCs elaborate a macromolecular cytokine that induces collagenase synthesis in skin fibroblasts and emphasize the importance of epithelial-stromal interactions in cutaneous tumor invasion. PMID- 2991392 TI - [Molecular weight and state of assembly of membrane proteins: an approach from measurement of low-angle laser light scattering]. PMID- 2991393 TI - [Animal cell cycle--G0 state]. PMID- 2991394 TI - [Organization and expression of genes encoded by transposons]. PMID- 2991395 TI - A clinicopathologic study of endometrial carcinoma with special reference to new histological variants. AB - The clinical and pathological features of 229 patients with endometrial carcinoma were analyzed with special reference to new histological variants. Histologically, 187 endometrial carcinomas were endometrioid form of adenocarcinoma, 10 mucinous carcinoma, 8 papillary serous carcinoma, 5 clear cell carcinoma, 1 secretory carcinoma, and 18 adenocarcinoma with squamous differentiation. Of these, papillary serous carcinoma was found to be a highly malignant form of endometrial carcinoma. Clear cell carcinoma was also associated with a poor prognosis. No fatal cases were observed in patients with mucinous carcinoma, secretory carcinoma, and adenocarcinoma with squamous differentiation. Patients with endometrioid form of adenocarcinoma were prognostically in between. Analysis of the patients with poor prognosis revealed that the length of time between onset of symptoms and surgery was not related to surviving periods and other prognostic factors such as clinical stage, histological grade, and myometrial invasion. PMID- 2991396 TI - [A clinicopathological study of 11 patients with Paget's disease of the vulva]. AB - A clinicopathological analysis of 11 patients with Paget's disease of the vulva was reported. A close association of this disease with cancer was strongly suggested in the histological examination. Nine of 11 patients had wide primary lesion; 4 patients had coexisting undifferentiated carcinoma (one patient with in situ sweat gland carcinoma) and one patient had squamous cell carcinoma in other lesion of the vulva. Five patients had downward growth of the tumor from the epidermis proper. There were, thus, only two patients who had Paget's cells confined within the epidermis. Three patients died of generalized metastasis of carcinoma and two patients had vulvar recurrence. The results of staining for PAS, PAS diastase resistant reaction, alcian blue, mucicarmin and carcinoembryonic antigen (CEA) were all positive in specimens with similar stainings to coexisting undifferentiated carcinoma of the vulva and metastasized lesions. CEA staining was positive in cytoplasm of Paget's cells and basal cells widely in the epidermis. It could be considered that the Paget's cells of the vulva originated multifocally from primitive stratum germinativum, which gave rise to the surface epithelium as well as all of the dermal appendages. PMID- 2991397 TI - [Studies on immunosuppressive acidic protein (IAP) in ovarian tumors]. AB - The value of IAP was measured in 64 patients with ovarioncus. The results obtained were as follows: The value of IAP in patients with malignant ovarioncus increased with the clinical progress of the disease. There was, however, no significant difference between the value in patients with malignant tumor in stage I or II and those with benign tumor. Therefore, the value has little usefulness for early diagnosis. In tissue typing of malignant ovarioncus, IAP was not positive in the case of intimoidal cancer, but it was almost on the same level in other cases. In terms of the relationship between IAP and plasma proteins or the immune index involved in various kinds of cancers, the value was closely correlated with the erythrocyte sedimentation rate and levels of CRP and mucoprotein, but not with the level of CEA, the LDH isozyme M/H ratio, or WBC and PPD skin reaction. Regardless of whether the tumor was benign or malignant, the value transiently increased postoperatively and then gradually decreased. Prognosis was poor in most cases with malignant ovarioncus, where the level of IAP remained high or rose again postoperatively. The level of serum IAP was almost correlated with the progress of the disease and is therefore useful as a marker in making a prognosis. PMID- 2991398 TI - [Clinical and roentgenological study of the hands in synbrachydactyly, constriction band syndrome and cleft hand complex]. AB - One hundred and six cases of synbrachydactyly, 63 cases of constriction band syndrome and 69 cases of cleft hand complex were clinically and roentgenologically studied and the following results were obtained. Synbrachydactyly is a group of hand anomalies in which the bone reduction appears transversely, and its severest form is a hand missing all the hand bones and the distal portion of the forearm, and with rudimentary digits on the stump. The association of dermal syndactyly or pectoralis muscle defect frequently seen in the milder cases of synbrachydactyly would be a secondary change to mesodermal hypoplasia or a related, but isolated malformation. Cleft hand complex which includes ordinary cleft hand, central ray polydactyly and central ray osseous syndactyly is a group of hand anomalies basically due to an ectodermal abnormality of separation of the digits. However, it is frequently associated with mesodermal hypoplasia or defect of various severity. The hand malformation in constriction band syndrome has a definite characteristic that the area of bone reduction is limited in phalanges. PMID- 2991399 TI - Ca+2 mobilization and fibrinogen binding of platelets refractory to adenosine diphosphate stimulation. AB - The mechanism of adenosine diphosphate (ADP)-induced refractoriness was explored with iodine 125-labeled fibrinogen and the fluorescent Ca+2 indicator quin-2 tetraacetoxymethyl ester (quin-2). Gel-filtered platelets were rendered refractory by incubation (30 minutes, 22 degrees C) with either 10 mumol/L ADP alone or ADP and 125I-labeled fibrinogen. During the incubation period, platelets incubated with ADP alone showed an initial increase in quin-2 fluorescence, which gradually returned to baseline levels. Addition of 125I-fibrinogen to aliquots of the platelet suspension at various times during incubation showed that fibrinogen binding was normal after 1 minute but decreased to 50% in 30 minutes. According to Scatchard analysis, this decreased binding was attributed to decreased fibrinogen receptor availability, not decreased receptor affinity. Moreover, similar numbers of glycoprotein (GP) IIb-IIIa complexes remained available on platelets before and after incubation, as judged by the ability of a monoclonal antibody (10E5) directed against a complex specific epitope on GPIIb or IIIa to bind to control and refractory platelets. After incubation, platelets aggregated poorly in response to restimulation with ADP, although the amount of fibrinogen they bound (50% of normal) was sufficient to aggregate control platelets. Platelet restimulation with ADP was not accompanied by a rise in quin-2 fluorescence or exposure of additional fibrinogen receptors. Stimulation of platelets with thrombin, however, led to a rise in quin-2 fluorescence, exposure of additional fibrinogen receptors, and enhanced aggregation. Restimulation of platelets with epinephrine also increased fibrinogen receptor exposure and restored the ability of platelets to aggregate, but was accompanied by barely detectable changes in quin-2 fluorescence similar to those observed with epinephrine-treated control platelets. Platelets incubated for 30 minutes with ADP and 125I-fibrinogen also showed an initial rise in quin-2 fluorescence, which returned to baseline levels during incubation, but the amount of platelet-bound fibrinogen, normal at the onset, remained quantitatively unchanged. Much of this fibrinogen, however, no longer dissociated from platelets in the presence of ethylenediaminetetraacetic acid or apyrase, suggesting that a different type of platelet-fibrinogen interaction had developed. Restimulation of these platelets with ADP was not accompanied by increased fibrinogen binding or quin-2 fluorescence and failed to elicit significant platelet aggregation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2991400 TI - Stimulus-specific neutrophil aggregation: evaluation of possible mechanisms for the stimulus-response apparatus. AB - We have assessed the effects of formyl peptides, leukotrienes, zymosan-activated serum (ZAS), and calcium ionophore A 23187 on the aggregation response of human neutrophils to characterize the mechanisms involved, particularly the possible contribution of the concomitant burst of oxidative metabolism and degranulation. We found that the tested stimuli induced aggregation waves highly characteristic for each agent. With formyl peptides, such as f-Met-Leu-Phe (fMLP), f-Met-Met-Met Met (fMMMM), and f-Nle-Leu-Phe-Nle-Tyr-Leu (fNLPNTL), peaks appeared after greater than 60 seconds and were followed by a slow disaggregation. In contrast, leukotriene B4 (LTB4) and its omega-oxidized metabolites, 20-OH-LTB4 and 20-COOH LTB4, induced rapidly occurring peaks that were maximal after 15 to 30 seconds and that were followed by a rapid and marked disaggregation. ZAS induced peak values after 30 to 90 seconds, and disaggregation kinetics were between fMLP and LTB4. A 23187 in high concentrations (10 to 50 mumol/L) elicited biphasic aggregation waves, whereas lower concentrations induced less pronounced and continuous aggregation responses. Superoxide dismutase, catalase, mannitol, and Tiron did not influence any of the aggregation characteristics. Colchicine and indomethacin did have some effects on peak values but not on the characteristic curve profiles, whereas cytochalasin B had a profound effect on the aggregation response for all tested stimuli. Neutrophils from a patient with chronic granulomatous disease exhibited aggregation responses similar to those of controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991401 TI - Human circulating monocytes internalize 125I-insulin in a similar fashion to rat hepatocytes: relevance to receptor regulation in target and nontarget tissues. AB - Circulating monocytes bind 125I-insulin in a specific fashion and have been used to analyze the ambient receptor status in humans. When freshly isolated circulating monocytes are incubated with 125I-insulin and examined by electron microscopic autoradiography, approximately 18% of the labeled material is internalized after 15 minutes at 37 degrees C. By 2 hours at 37 degrees C, approximately one half of the 125I-insulin is internalized. Internalization occurs also at 15 degrees C but at a slower rate. Furthermore, the monocytes bind and internalize 125I-insulin in a manner that mirrors that of major target tissues, such as rat hepatocytes. These data suggest that the insulin receptor of the circulating monocyte might be regulated by adsorptive endocytosis in a manner analogous to that of target tissue, such as the liver. PMID- 2991403 TI - Elimination of silica gel from gangliosides by using a reversed-phase column after preparative thin-layer chromatography. AB - A simple and effective procedure has been developed for eliminating silica gel from gangliosides after preparative thin-layer chromatography. Gangliosides were extracted from the scraped silica gel with chloroform-methanol-water 10:10:3 (by volume) and dried. The residue was suspended in 0.1 M NaCl and centrifuged. After filtration through a sintered glass funnel, the supernatant was applied to a reversed-phase column. The column was washed with 0.1 M NaCl, water, and methanol water 1:1 (v/v). Gangliosides were eluted from the cartridge with methanol with yields of 92% and 90% in terms of dry weight and sialic acid content, respectively. Almost all of the silica gel contamination was eliminated from gangliosides by this procedure. PMID- 2991402 TI - Extra-nasopharyngeal extension of juvenile angiofibroma. AB - Nasopharyngeal angiofibroma (NPA) is a highly vascular, infiltrative tumour that occurs exclusively in the nasopharynx of young males. It is of special interest on account of its serious complications, and the difficulties that are encountered at the time of treatment. Various surgical approaches have been mentioned for both naso- and extra-nasopharyngeal angiofibromas. But a massive extra-NPA involving the naso-spheno-oro-palato-maxillary-infratemporal region, orbit and cheek is difficult to treat adequately with any of the approaches so far described in the literature. In these cases, a more radical approach (constituting embolization and subsequent clamping or ligation of the external carotid artery followed by total maxillectomy) may be needed. These procedures will not only give a dry operative field but also adequate exposure to the different parts of the extension of the tumour, thereby facilitating complete removal without any risk of recurrence. PMID- 2991404 TI - Experimentally-induced jaw sarcoma in the rat. AB - Jaw bones differ embryologically from limb bones and the axial skeleton as they are derived from migrating neural crest cells and are not preformed in cartilage as are the latter bone types. The purpose of the present study was to determine whether this embryologically different nature implied a different response to a locally applied carcinogenic compound, 4-hydroxyamino-quinoline-1-oxide; this within the context of elucidating why human jaw osteo- and fibrosarcomas behave differently from identical tumours elsewhere in the skeleton. Rat mandibles were exposed to the carcinogen. Of the 5 rats used, 3 developed tumours which were however histologically identical to the tumours experimentally induced in the limb bones by the same carcinogenic agent. As a consequence, it is concluded that the embryologically different nature of the jaw bone does not imply that local exposure to a carcinogenic agent induces other kinds of tumours as are seen in the extragnathic skeleton. PMID- 2991405 TI - Malignant fibrous histiocytoma of the maxilla. Case report and review of the literature. AB - A case of malignant fibrous histiocytoma (MFH) occurring in the maxilla is presented. Although MFH is now recognised as the most common soft tissue sarcoma of late adult life, it rarely occurs in bone and then more usually in the long bones. Presentation affecting the facial bones and skull is extremely rare, but should be considered in the differential diagnosis of bone tumours affecting the maxilla. Skeletal lesions appear more aggressive than those in soft tissues. Early radical excision is the treatment of choice although radiotherapy for inoperable tumours may lead to regression, as in the case presented here. PMID- 2991407 TI - Liver angiotensin II receptors in the rat: binding properties and regulation by dietary Na+ and angiotensin II. AB - Recent evidence suggests that angiotensin II (AII) acts on the liver via specific receptors. The aims of this study were to examine the general binding properties of these receptors in the rat and to determine the role of dietary Na+ and AII in the regulation of AII receptors. Binding of 125I-labelled Ile5-AII to liver membranes was saturable and behaved as a single class of sites with an affinity (Ka) of 2.9 l/nmol and a Hill coefficient of 0.99. Kinetic analysis of AII binding gave estimates for the rates of association and dissociation of 42 l/mumol per min and 1.5 X 10(-2)/min respectively. The binding of analogues exhibited the following order of potency: Val5-AII greater than Ile5-AII greater than AIII greater than Sar1-Ala8-AII greater than Sar1-Gly8-AII greater than AI greater than des-Asp-AI greater than C4-C8 pentapeptide greater than Phe1-Tyr8 AII greater than neurotensin greater than luteinizing hormone-releasing hormone. Binding was enhanced by Mg2+ and Ca2+ and inhibited by EDTA and the reducing agent dithiothreitol. Low dietary Na+ affected in a biphasic manner both the Ka and concentration (Ro) of liver AII receptors. Initially, Ro decreased from 25.9 +/- 3.8 (control) to 16 +/- 1.9 (S.E.M.) pmol/g tissue by 1.5 days but thereafter it rapidly increased to 47.3 +/- 8.7 pmol/g tissue (3.5 days) and remained elevated to the end of the experiment, 8.5 days later. The Ka initially increased from 2.7 +/- 0.3 (control) to 4.3 +/- 0.5 l/nmol (1.5 days) and then decreased steadily to 1.2 +/- 0.1 l/mol (8.5 days).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991406 TI - Catechol oestradiol control of FSH-stimulated granulosa cell steroidogenesis. AB - Catechol oestrogens are formed in the ovary but it is not known if they have a local function. Working with primary granulosa cell cultures prepared from immature rat ovaries, we found that the presence of 2-hydroxyoestradiol in the culture medium (48 h incubation) dramatically enhanced the steroidogenic response (progesterone production) to human FSH (100 ng/ml). The effect of 2 hydroxyoestradiol was dose-dependent and maximal (approximately 40 times the response to FSH alone) at 3.0 microM. The stimulatory action of 1.0 microM 2 hydroxyoestradiol was greater than 10 times more than that of 1.0 microM oestradiol but only half that of 1.0 microM testosterone; other catechol oestrogens (2-hydroxyoestrone, 4-hydroxyoestradiol, 2-methoxyoestradiol and 2 methoxyoestrone) were not stimulatory. The stimulatory actions of 2 hydroxyoestradiol and testosterone were partially additive and each was antagonized in the same way by the presence of a specific antiandrogen (SCH16423). These observations suggest a role for intrafollicular catechol oestradiol in modulating FSH-stimulated granulosa cell steroidogenesis; its mechanism of action may be similar to that of testosterone. PMID- 2991408 TI - Inhibitory effect of colchicine and vinblastine on transport of glucagon receptors to the plasma membrane in cultured rat hepatocytes. AB - The role of microtubules in the regulation of glucagon receptors on cultured rat hepatocytes was studied. Antimicrotubular reagents, colchicine and vinblastine, did not affect the binding of 125I-labelled glucagon to hepatocytes at 4 degrees C. At 20 and 37 degrees C, however, the reagents reduced the binding after 60 or 90 min of incubation. Scatchard analysis indicated that the reduction in the binding was due to loss of glucagon-receptor populations. If hepatocytes were preincubated with both unlabelled glucagon and the reagents at 37 degrees C, the binding of the ligand to the cells decreased markedly after a certain delay. The reagents did not inhibit the internalization of the ligand in the cells until 30 min of incubation at 37 degrees C. The results suggest that the microtubule system plays a role in the transport of glucagon receptors to the plasma membrane, which is followed by their internalization. PMID- 2991409 TI - Measurement of low concentrations of bovine thyrotrophin by iodide uptake and organification in porcine thyrocytes. AB - Thyrocytes isolated from porcine thyroids by mechanical and enzymatic dispersion and cultured in Eagle's minimal essential medium, supplemented with 5% (v/v) fetal calf serum, glutamine and cortisol, formed a continuous monolayer within 48 h. This monolayer was without cytochemical peroxidase and diaphorase (NADPH reoxidation) activity. In the presence of bovine thyrotrophin (bTSH; 50 mu./l) the cells developed a follicular-like architecture which was maximal at 4 days before reverting back to a uniform monolayer at 6 days. There were no detectable changes in the total DNA content over this period. The follicular structures had marked diaphorase and peroxidase activity, the latter being apically distributed. Concomitant with follicle formation bTSH induced uptake and organification of iodide presented to the cells during the last 6 h of culture. The extent of this process depended on the dose of bTSH and the duration of stimulation. The most sensitive effects for both iodide uptake and organification occurred with 1 mu. bTSH/l and were maximal with 100 mu./l. Uptake and organification were increased 20 +/- 8-fold and 9.6 +/- 2-fold (n = 10) respectively over the control with 100 mu./l and the doses of bTSH at which a half maximal response was seen (ED50) were 15 +/- 2 and 7 +/- 1 (S.D) mu./l (n = 10) respectively. On changing the culture medium to a serum-free system using HB101 culture medium the stimulation time for the most sensitive bTSH effect was reduced to 2.5 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991410 TI - Salmonid melanin-concentrating hormone inhibits corticotrophin release. AB - Melanin-concentrating hormone (MCH) is a neural peptide associated with colour change in fishes. We show here that it also inhibits corticotrophin (ACTH) secretion. Synthetic salmonid MCH at a concentration of 100pmol/l reduced the in vitro release of ACTH by pars distales (pDs) taken from stressed trout. At lower concentrations (10pmol/l) the peptide inhibited CRF-41-induced secretion of ACTH by pDs removed from unstressed trout, while at higher concentrations (10nmol/l) it reduced the corticotrophic response of rat pituitary tissue to CRF-41. PMID- 2991411 TI - Possible role of uterine contractions in the short-term fluctuations of plasma ACTH concentration in fetal sheep. AB - Uterine contractions, induced by the administration of oxytocin to sheep between d 123-144 of pregnancy, were associated with a mean transient decrease in fetal PaO2 of 2.8 mm Hg within 5 min. These changes were associated with a rapid increase in the concentration of ACTH in fetal plasma. There was a significant (P less than 0.05) increase in the percentage change (+40 to +47%) over basal ACTH levels in fetal plasma at +5, +15 and +20 min after oxytocin. Administration of saline had no significant effect on intrauterine pressure, fetal PaO2 or fetal plasma ACTH levels. We speculate that increases in uterine activity and/or transient decreases in fetal PaO2 may contribute to short-term fluctuations in plasma ACTH in fetal sheep. PMID- 2991412 TI - Repeated thyrotrophin stimulation of thyroid secretion: lack of refractoriness in vivo. AB - It has been reported that prior exposure of thyroid tissue to TSH in vitro induces a state of refractoriness to new challenges of the hormone. We have investigated the effect of repeated TSH treatment on thyroid secretion to determine whether such refractoriness exists in vivo. The rate of thyroid secretion was estimated by measuring the rate of hydrolysis of labelled thyroglobulin from mouse thyroid glands in vitro. The thyroid glands were labelled in vivo with 131I and then cultured for 20 h in the presence of mononitrotyrosine, an inhibitor of iodotyrosine deiodinase. The rate of hydrolysis of labelled thyroglobulin was measured as the percentage of radioactivity released as free iodotyrosines and iodothyronines into the gland and the medium at the end of incubation. Thyrotrophin was administered in vivo at hourly intervals for 2-4 injections. The corresponding control group received saline injections every hour except for the last injection when they received TSH. The peak rates of thyroglobulin hydrolysis, measured 2 h following the last injection, were similar in animals receiving two, three or four TSH injections and were not different from those in the control groups. Serum triiodothyronine and thyroxine concentrations 2 h after the last injection were higher in the groups receiving multiple TSH injections. Thyroidal cyclic AMP accumulation in response to TSH was markedly depressed in the group receiving multiple injections compared with the group receiving a single injection of TSH in vivo.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991413 TI - Peroxidase-catalysed iodotyrosine formation in dispersed cells of mouse extrathyroidal tissues. AB - A method has been developed for the isolation of cells, high in iodine uptake and peroxidase activity, from the stomach and submaxillary gland of mice. The isolated cells could produce protein-bound monoiodotyrosine, di-iodotyrosine and an unknown iodocompound. The reactions were catalysed by peroxidase and were sensitive to antithyroid drugs and haemoprotein inhibitors but were insensitive to TSH. In-vitro iodination of stomach or submaxillary soluble proteins with the respective peroxidase yielded similar iodocompounds while thyroxine was produced when thyroglobulin was used instead. PMID- 2991414 TI - Endocrine control of uterine oxytocin receptors in the ewe. AB - Specific binding of [3H]oxytocin to high affinity sites (hormone receptors) in membrane preparations from uterine tissues of the ewe has been determined at varying stages of the oestrous cycle and in pregnancy. Mean receptor concentrations in caruncular and intercaruncular endometrium and in myometrium were 14.2, 1.9 and 13.0 fmol/mg protein respectively between days 10 and 13 of the cycle. By the day of oestrus these values had increased to 749, 1085 and 179 fmol/mg protein. These increases in receptor concentrations coincided with luteolysis and falling plasma progesterone levels and followed the preovulatory decline in peripheral oxytocin and rise in ovarian venous oestradiol-17 beta. Receptor concentrations were low in all uterine tissues from pregnant animals between days 14 and 19 after oestrus. Analysis of binding parameters by Scatchard plot suggested a single population of receptor molecules in each of the tissues studied with apparent dissociation constants in the range 1.9-2.2 nmol/l. A number of naturally occurring neurohypophysial peptides inhibited binding of [3H]oxytocin to the receptor from ewes at oestrus; the cross-reactions of arginine vasopressin and vasotocin exceeded that of oxytocin. Use of a receptor binding assay to measure oxytocin in extracts of corpora lutea on days 4 and 10 after oestrus gave values similar to those obtained by radioimmunoassay, suggesting the absence of other receptor-active peptides in the corpus luteum. It is concluded that the oxytocin receptor is present in both components of the endometrium, as well as in the myometrium and that changes in uterine receptor concentrations before oestrus are consistent with receptor activation by steroid hormones. PMID- 2991415 TI - Major histocompatibility complex restriction fragment length polymorphisms define three diabetogenic haplotypes in BB and BBN rats. AB - Class I and II major histocompatibility complex (MHC) probes can be used to subdivide diabetes-prone BB rats and their BBN control strain, coderived from the same outbred colony by selection against diabetes. Class II probes (A-alpha in particular) distinguish four restriction fragment length polymorphisms (RFLP), termed 1a, 1b, 2a, and 2b, in the BBN population, only one of which (2a) is found in BB rats. The degree of class II RFLP in the population studied is RT1.B-alpha greater than or equal to RT1.B-beta greater than RT1.D-alpha greater than or equal to RT1.D-beta, suggesting that intra-class II region dynamics may be different in rats compared with mice. A class I probe (S16) absolutely distinguished BB from BBN rats, since all BB rats exhibit an RFLP pattern termed 2a0, while 2a BBN rats can be subdivided into 2a1 and 2a2 forms. Serologic evaluation has shown that 2a0, 2a1, and 2a2 rats express RT1.AuBu, 1a rats express RT1.AaDa, and 1b rats express neither RT1a nor RT1u at the loci tested. A breeding study was carried out to determine the diabetogenicity of the MHC defined RFLP's. As expected, the BB-derived 2a0 is diabetogenic. The BBN-derived 2a1 and 2a2 RFLPs are also diabetogenic, while 1a and 1b rats do not carry MHC linked diabetogenic genes. The MHC-linked diabetes gene acts in a functionally recessive manner, since there is a 10-fold higher incidence in homozygotes than in heterozygotes. Analysis of the RFLP patterns leads us to hypothesize that the 2a1 RFLP results from a crossover between 1a and 2a0 MHCs and that the diabetogenic MHC-linked gene is on the class II side of Qa and T1. The availability of three diabetogenic MHC haplotypes should help localize the MHC linked diabetogenic gene of rats. PMID- 2991417 TI - Glucocorticoid-stimulated biosynthesis of low density lipoprotein receptor in cultured fibroblasts. AB - Preincubation of human skin fibroblasts in the presence of 10(-6)-10(-5) mol/l glucocorticoids (dexamethasone) causes a concentration and time-dependent increase of receptor-mediated internalisation of [125I]LDL. This increase is due to a glucocorticoid-specific stimulation by 40-50% of LDL receptor synthesis as demonstrated by an increased incorporation of [35S]methionine into immune precipitated receptor protein. In contrast the rate of synthesis of total cell protein and of lysosomal cathepsin D is not significantly influenced by dexamethasone. The increased LDL receptor synthesis is accompanied by an enhanced synthesis of cholesterol from [2-3H]mevalonolactone and [1-14C]acetate. The glucocorticoid-induced enhancement of LDL receptor and cholesterol synthesis is abolished by preincubation of the cells with dexamethasone in combination with 25 hydroxycholesterol. PMID- 2991418 TI - Some effects produced by isoprenaline upon neuromuscular transmission in mice. PMID- 2991416 TI - Allorestricted cytotoxic T cells. Large numbers of allo-H-2Kb-restricted antihapten and antiviral cytotoxic T cell populations clonally develop in vitro from murine splenic precursor T cells. AB - Cytotoxic T lymphocyte (CTL) responses of splenic T cells from C57BL/6 B6) mice and mutant H-2Kbm1 (bm1) mice to haptenic (trinitrophenyl [TNP] ) and herpes simplex virus (HSV) determinants in the context of an allogenic (wild-type or mutant) H-2Kb molecule were analyzed in a modified limiting dilution system. In the B6-anti-bm1TNP mixed leukocyte reaction (MLR), estimated frequencies for precursors of CTL clones that lysed bm1TNP targets ranged from 1/120 to 1/400; in the bm1-anti-B6TNP MLR, estimated frequencies of precursors of CTL clones that lysed B6TNP targets ranged from 1/500 to 1/1,300. Estimated frequencies for precursors of CTL clones that lysed the respective unmodified and TNP-modified allogeneic targets were two- to three-fold lower. Lytic specificity patterns determined by split-well analysis showed that at least 20-30% of the generated CTL populations (selected for a high probability of clonality) in both MLR displayed allorestricted lysis of TNP-modified concanavalin A blast targets. In the B6-anti-bm1HSV MLR, estimated frequencies for precursors of CTL clones that lysed bm1HSV targets ranged from 1/70 to 1/300; in the bm1-anti-B6HSV MLR, estimated frequencies for precursors of CTL clones that lysed B6HSV targets ranged from 1/300 to 1/1,200. Again, estimated frequencies for precursors of CTL clones that lysed the respective noninfected and virus-infected allogeneic targets were two- to fourfold lower. Of the CTL populations selected for a high probability of clonality at least 30-60% displayed allorestricted lysis of virus infected lipopolysaccharide blast targets in both MLR. It is concluded that a large fraction of clonally developing CTL populations stimulated with TNP modified or HSV-infected allo-H-2Kb-bearing cells displayed an allorestricted pattern of recognition. It was further evident that the estimated frequencies of splenic precursors that generated allorestricted CTL clones was two- to threefold higher than the estimated frequencies of precursors that gave rise to the respective alloreactive CTL populations. PMID- 2991419 TI - Study on enterovirus 70: age distribution of neutralizing antibody and some characteristics of the virus. PMID- 2991420 TI - TSH receptor in thyroid neoplasms. PMID- 2991421 TI - Evolution of Tn21-related transposons: isolation of Tn2425, which harbours IS161. AB - The isolation of two multi-resistance transposons, Tn2425 and Tn1831, and their relation to Tn21 and Tn2424, is described. A 1.7 kb segment present in Tn2424 and Tn2425 was identified as an IS element by rec-independent transposition, resulting in a cointegrate structure that carries two direct repeated copies of the IS element. By the isolation of this IS element we demonstrated that transposition is one mechanism leading to sequence variations in Tn21-like structures, especially in the region between the mer operon and the sul gene. PMID- 2991422 TI - Plasmid fusions mediated by one end of TnA. AB - We have observed plasmid fusions in a recA background mediated by a single end of TnA. These occur when transposase is provided either in cis or in trans. Insertions of the plasmid carrying the TnA inverted repeat sequence occur at many sites in the target plasmid. The point of fusion on the plasmid carrying TnA sequences always appears to be located in the region which carries the TnA inverted repeat sequence. In contrast to the transposition of an intact TnA element, plasmid fusions mediated by one end of TnA are very rare events. The implications of our results for models of transposition are discussed. PMID- 2991424 TI - The genome of Bacillus subtilis phage SPP1: structure of an early promoter. AB - The strongest of five 'early' promoters of Bacillus subtilis phage SPP1 was localized in a DNA restriction fragment by analysis of RNA polymerase binding and R-loop formation. The nucleotide sequence of the promoter region was established. The signal structures identified were similar to those recognized by the sigma 55 RNA polymerase of B. subtilis. The promoter precedes an open reading frame with 51 codons. A protein with the Mr predicted from the nucleotide sequence was identified in minicells. PMID- 2991423 TI - Structural organization of a hypermethylated nuclear DNA component in Physarum polycephalum. AB - Digestion of Physarum polycephalum nuclear DNA using the restriction endonuclease HpaII generates two components, distinguishable on the basis of their molecular size. The high-molecular-weight, HpaII-resistant component, which accounts for 20% of the DNA, contains a fivefold greater concentration of 5-methylcytosine residues than the low-molecular-weight HpaII-digested fraction. Segments of hypermethylated (M+) DNA are largely composed of a single, long, highly repeated sequence, and this major element is sometimes associated with other less highly repetitive sequences in the M+ DNA fraction. Restriction mapping of cloned Physarum M+ DNA segments, and Southern blot analysis of genomic DNA using subcloned segments of M+ DNA as a probe, provide evidence for sequence variation within different copies of the dominant highly repeated element, and possibly the other associated repeats in M+ DNA, and additionally that almost complete tandemly repeated copies of the major repeat are found in some M+ DNA segments. PMID- 2991425 TI - Chronic shedding of bovine enteric coronavirus antigen-antibody complexes by clinically normal cows. AB - Using an ELISA for the detection of virus-specific immune complexes, ten cows were found to be shedding bovine enteric coronavirus. The shedding patterns from five of these animals were followed for a period of 12 weeks, and all were found to be chronically shedding virus. Despite the presence of both faecal and serum antibody the infection was not cleared; therefore, the role of cell-mediated immunity (CMI) was investigated by immunosuppressing the chronically shedding cows with dexamethasone. No major role for CMI in maintaining the chronic infection could be determined, although immunosuppression did result in a temporary reduction in the shedding of virus-specific immune complexes. PMID- 2991426 TI - Cell cycle position and expression of encephalomyocarditis virus in mouse embryo fibroblasts. AB - Infection of mouse embryo fibroblasts in G1 or S phase with encephalomyocarditis virus gave different kinetics of viral RNA synthesis. In S phase cells, RNA synthesis was faster and reached higher levels than in G1 cells. Virus-specified proteins were fewer in G1 cells than in S cells during the early stage of the infection and c.p.e. in G1 cells appeared about 4 h later than in S cells. Addition of a cellular factor with ability to affect cell conformation had an inhibitory effect on viral RNA synthesis. PMID- 2991427 TI - Temperature sensitivity of the transcriptase of mutants tsB1 and tsF1 of vesicular stomatitis virus New Jersey is a consequence of mutation affecting polypeptide L. AB - Two conditional transcriptase-negative mutants of vesicular stomatitis virus (VSV) serotype New Jersey, tsB1 and tsF1, their revertants tsB1/R1 and tsF1/R1 and the wildtype virus were dissociated into pellet, NS and L fractions and, after reconstitution of these in various combinations, the transcriptase activities were assayed in vitro at the permissive (31 degrees C) and restrictive (39 degrees C) temperatures. The pellet fractions contained the virion RNA polypeptide N complexes, while the NS and L fractions were essentially pure preparations of these polypeptides. The synthesis of RNA by the reconstituted pellet and L fractions was inhibited at 39 degrees C only when the L fractions of tsB1 or tsF1 were used. Addition of the NS fractions to the reconstituted pellet and L fractions did not alter the rates of RNA synthesis. These results demonstrate that polypeptide L is the temperature-sensitive polypeptide of both mutants tsB1 and tsF1 and support previous observations that polypeptide L is the transcriptase itself. The fact that a second mutant of complementation group F, tsF2, is transcriptase-positive but replicase-negative suggests that polypeptide L is involved both in transcription and replication. Intracistronic complementations may account for the observation that the temperature-sensitive mutations affect polypeptide L in complementation groups B and F. PMID- 2991428 TI - The physical state of human papillomavirus type 16 DNA in benign and malignant genital tumours. AB - Cloned DNA from human papillomavirus (HPV) type 16 was subjected to restriction enzyme analysis. A genome size of 7.8 +/- 0.1 kb was determined and restriction maps were prepared. Fragments of HPV 16 DNA were nick-translated and hybridized with fragments of HPV 6b DNA. The two genomes appeared to be colinear. The physical state of HPV 16 DNA in genital tumours was analysed. In each of six benign tumours the viral DNA was detected exclusively as 8 kb circles. In four malignant tumours the viral DNA appeared to be integrated within the host genome but one cervical carcinoma and one case of Bowen's disease also contained oligomeric episomal molecules of viral DNA. One cervical carcinoma (WV 2965), containing only integrated viral DNA, was examined in detail. HPV 16 DNA was integrated as head-to-tail tandem repeats at more than one site. Three virus/cell junction fragments from this tumour were cloned. Two contained lengths of repetitive cellular DNA and one a length of apparently single copy cellular DNA. PMID- 2991429 TI - Association of type I DNA topoisomerase with herpes simplex virus. AB - A topoisomerase activity is associated with herpes simplex virus type 1. The enzyme was recovered from purified virions which were disrupted with 6 M guanidine-HCl followed by renaturation of extracted proteins. Based upon the following observations, the virion activity is classified as a type I topoisomerase: (i) the linking number of a unique DNA topoisomer is altered in steps of one; (ii) ATP and MgCl2 are not required for activity; (iii) the enzyme can be trapped in a covalent complex with DNA; (iv) the covalent linkage to DNA is through a 3' phosphoryl bond. A number of lines of evidence strongly indicate that the topoisomerase is external to the nucleocapsid. For example, the activity was released by treatment of intact virions with NP40, and subsequent washing steps extracted most residual activity. When guanidine extracts were prepared from nucleocapsids, topoisomerase activity was not detectable. Finally, DNA within the virion did not appear to contain covalently attached proteins with properties similar to topoisomerases. Thus, the enzyme appears to be a component of the envelope or tegument structure of the virion. PMID- 2991430 TI - A herpes simplex virus type 1 mutant with a deletion in the polypeptide-coding sequences of the ICP4 gene. AB - A deletion mutant derived from herpes simplex virus type 1 (HSV-1) strain ANG was analysed. The deletion mapped within the polypeptide-coding region of the immediate-early ICP4 gene. Based on DNA sequence data the deletion was shown to comprise 84 base pairs. In the wild-type genome of strain ANG these sequences were almost completely homologous to the known sequences of HSV-1 strain 17. The ICP4 polypeptide induced by the mutant was similar in size to the wild-type ICP4 protein and was recognized by a monoclonal antibody against ICP4. The data presented suggest that the deletion corresponds to a region on the ICP4 polypeptide that is nonessential for the replication of the virus in vitro. PMID- 2991431 TI - Epstein-Barr virus nuclear antigen (EBNA): size polymorphism of EBNA 1. AB - The mol. wt. of the polymorphic Epstein-Barr virus (EBV) nuclear antigen (EBNA) molecule (EBNA 1) encoded by the BamHI K fragment of the EBV DNA has been determined in 14 EBV-carrying lymphoblastoid and Burkitt's lymphoma cell lines. There is no obvious correlation between the size of this polypeptide and any properties of the cells from which it is derived, other than those related to the strain of transforming virus. We confirm that the polymorphic region of this molecule is the glycine-alanine copolymer encoded by the third internal repeat of the EBV genome (IR3) and we consider the significance of this domain. PMID- 2991432 TI - Inhibition of herpes simplex virus type 1 penetration by cytochalasins B and D. AB - Internalization of herpes simplex virus type 1 (HSV) KOS strain by HEp-2 cells was reversibly inhibited by pretreatment of cells with cytochalasins B and D. Internalization of virus following preincubation at 4 degrees C and temperature shift to 37 degrees C was normally preceded by a 5 to 8 min lag period and was complete within 20 to 30 min. A similar lag period followed HSV addition at 37 degrees C. Cytochalasin D was fivefold more active on HSV entry than cytochalasin B, with 50% inhibition at 2 microM and 10 microM respectively. Inhibition was completely reversible, such that all cell-bound infectious virus was recovered upon removal of cytochalasin. In conjunction with previous reports, the activity of cytochalasin on HSV entry suggests that a change in cytoskeletal structure following virus attachment triggers a microfilament activity important for internalization of HSV by HEp-2 cells. PMID- 2991433 TI - A genetic probe for identifying bluetongue virus infections in vivo and in vitro. AB - We have used a DNA copy of segment 3 RNA of bluetongue virus serotype 17 (BTV-17) to detect sequence homology among the equivalent segments of five U.S.A. BTV serotypes (BTV-2, BTV-10, BTV-11, BTV-13 and BTV-17) as well as 14 other BTVs isolated from different endemic areas of the world. Both by in situ and Northern hybridization all the BTV serotypes were found to have RNA that reacted with the DNA probe. No homology was detected with epizootic haemorrhagic disease virus serotype 1, a related orbivirus. The BTV-17 DNA clone has also been used to detect viral RNA in infected sheep blood. This information has led us to develop a simple and sensitive procedure for the detection of viral genome-biotinylated clone DNA hybrids in vivo or in cultured cells following direct staining with either the avidin-fluorescein complex or the streptavidin-horseradish peroxidase complex. PMID- 2991434 TI - Viral aetiology of adult T-cell leukaemia. PMID- 2991435 TI - Comparative studies of A/Leningrad/134/57 wild-type and 47-times passaged cold adapted mutant influenza viruses: oligonucleotide mapping and RNA-RNA hybridization studies. AB - Direct biochemical evidence for the existence of mutations in five of the RNA segments of the A/Leningrad/134/57 cold-adapted 47th passage mutant as compared with its wild-type progenitor has been obtained using two techniques. T1 oligonucleotide mapping of total viral RNA as well as of individual RNA segments revealed changes in RNAs 4, 5 and 6. Analysis of S1 nuclease-treated RNA-RNA hybrids on polyacrylamide gels revealed changes in at least one of the polymerase genes as well as in RNAs 4, 5, 6 and 7. These findings provide a direct demonstration for the existence of multiple mutations in the cold-adapted mutant vaccine strain. PMID- 2991436 TI - Variation in susceptibility to avian sarcoma viruses and expression of endogenous avian leukosis virus antigens in specific pathogen-free chicken lines. AB - Five lines of chickens maintained as specific pathogen-free flocks in Australia were characterized in relation to endogenous antigens and endogenous avian leukosis virus expression. Embryos of line N were predominantly of C/E phenotype, uniformly positive for group-specific antigen and chick helper factor (gs+chf+) and 38% expressed endogenous virus at a very low titre. Embryos of line M4 were uniformly of C/ABE phenotype and were either gs+chf+ or gs-chf+. Line W19 embryos segregated for susceptibility to viruses of subgroup A, B and D and were either of C/E or C/ABE phenotype. The majority of W19 embryos were gs+chf+ with a small proportion being gs+chf-. Line I13 embryos were either of C/0 or C/ABE phenotype, uniformly gs-chf- and 44% of embryos expressed endogenous virus at a low titre. Line S segregated for susceptibility to subgroup E virus and embryos were either of C/E or C/0 phenotype, while the majority of embryos from line S were gs-chf- with some embryos being gs+chf+ or gs-chf+. The degree of interference of gs+chf+ and gs-chf+ phenotypes with subgroup E virus infection was identical with the interference patterns of classical gs+chf+ and gs-chf+ phenotypes. The resistance to infection with avian sarcoma viruses of subgroups E in lines N and M4, and to a degree in line W19, was highly associated with the presence of chf. Resistance to subgroup E virus was independent of chf in lines S and I13, probably being under the control of an independent locus. Cellular restriction of endogenous virus replication existed in all subgroup E virus-susceptible cells of line I13 in contrast to cells of line S which supported replication of endogenous virus. The phenotype of chicken cells for the expression of endogenous gs antigen and chf could accurately be predicted from the test performed on whole blood cells. PMID- 2991437 TI - Comparative expression of the hepatitis B surface antigen gene in biochemically transformed human, simian and murine cells. AB - In this paper, we show that the pattern of expression of the human hepatitis B surface antigen (HBs Ag) gene, transfected along with a dominant selectable marker into mammalian cells, is complex. In human hepatoma (HepG2) cells, late transient expression occurs and permanent expression takes place at high frequencies in the selected clones. In HeLa and human xeroderma pigmentosum (GM4312A)-derived cells, the late transient expression is barely seen or absent and permanent expression is only seen in a few selected clones. In monkey kidney Vero cells, late transient expression has been described and we show in this report that only 5% of the selected clones are capable of expressing HBs Ag in a permanent manner. In most of the Vero clones, the absence of HBs Ag expression is mainly due to HBs Ag gene rearrangements. We have selected and amplified more than 500 transfected Vero clones and have characterized in detail one clone (GAR1412) which is a permanent high-level HBs Ag expressor. PMID- 2991438 TI - Effect of tacaribe virus infection on host cell protein and nucleic acid synthesis. AB - Tacaribe virus stocks were prepared which induced definite lytic responses in Vero cells infected at multiplicities giving synchronous infection. Under these conditions, the first signs of cytopathic effect (c.p.e.) appeared at about 30 h post-infection and cell lysis occurred after 40 h. Before the onset of cytopathic changes, the virus induced inhibition of host cell protein, DNA and RNA (primarily rRNA) synthesis. These were designated c.p.e. (+) virus stocks. The effect of virus on host cell macromolecular synthesis and development of c.p.e. were not related to the virus isolate, but to the conditions under which the virus was produced. Thus, from a single virus clone, working stocks were derived which could or could not induce inhibition of host cell functions and c.p.e. development. The virus stocks that did not induce inhibition are defined as c.p.e. (-). Analysis of [3H]leucine-labelled proteins from Vero cells infected with either the c.p.e. (+) or the c.p.e. (-) virus stocks revealed synthesis of two virus-specific polypeptides migrating with mobilities corresponding to mol. wt. 68 000 and 79000. These are presumed to correspond, respectively, to the nucleoprotein and to the minor polypeptide p79. In cells infected with the c.p.e. (+) virus stock, the virus-specific polypeptides were synthesized at times when there was a drastic inhibition of host cell protein synthesis. The yield of infectious progeny during the first 24 h of infection is similar in Vero cells infected with either the c.p.e. (+) or the c.p.e. (-) virus stocks. The proportion of defective interfering particles was much higher in the c.p.e. (-) than in the c.p.e. (+) virus stocks. The results presented here are the first demonstration that an arenavirus affects the biosynthetic machinery of the host cell. PMID- 2991439 TI - Retrieval of latent herpes simplex virus type 1 genetic information from murine trigeminal ganglia by superinfection with heterotypic virus in vivo. AB - Mice previously latently infected with the F strain of herpes simplex virus type 1 (HSV-1) can be successfully colonized with a second virus strain if HSV-2 is introduced at the same peripheral site as HSV-1. On the other hand, HSV-1 strains seemed able mutually to exclude establishment of latency with each other. Mice (3 months or 3 years after nasal infection) latently infected with HSV-1 were thus superinfected with HSV-2. The mice were sacrificed 2 days post-infection when HSV 2 replication in the ganglia was found to have commenced. Ganglia were homogenized immediately and virus was plaqued on permissive cells. HSV-1 plaques were regularly obtained among HSV-2 plaques as assessed by staining with an enzyme-linked immunosorbent using a type-specific monoclonal antibody recognizing glycoprotein C of HSV-1. DNA from this virus had identical restriction endonuclease patterns (EcoRI, BamHI and HindIII) to the F strain used to infect the animals latently. HSV-1 was not retrieved from ganglia of controls superinfected with a neuroadapted vaccinia virus or were mock-superinfected. The results suggest that it is possible to superinfect a latently infected ganglionic neuronal cell with a heterotypic HSV strain and that the subsequently introduced HSV-2 can act in trans to induce reactivation of latent HSV-1. PMID- 2991440 TI - The effect of triterpenoid compounds on uninfected and herpes simplex virus infected cells in culture. I. Effect on cell growth, virus particles and virus replication. AB - The related triterpenoid compounds carbenoxolone sodium (CBX) and cicloxolone sodium (CCX) have been investigated in clinical trials for treatment of herpes simplex virus (HSV) infections. When the drugs were tested in vitro, two dose related effects on BHK cells became apparent: the rate of cell growth was reduced and the drugs exhibited cytotoxicity at high concentrations. Flow 2002 cells, in contrast, were apparently unaffected by all drug concentrations tested. The effect of up to 3 days incubation with 100 microM-CCX on BHK cells was reversible. The presence of 500 microM-CBX or 300 microM-CCX during the HSV replication cycle reduced the infectious virus yield to less than 0.01%: CCX is the more potent anti-herpes agent. The contribution made by cytotoxicity to the overall antiviral effect (measured by 24 h yield) was negligible in Flow 2002 cells, and was relatively unimportant in BHK cells. The amount of HSV-1 or HSV-2 adsorbing to pretreated BHK cells was reduced by 20% and 40% respectively at the highest drug concentrations. Neither 500 microM-CBX nor 300 microM-CCX treatment for 24 h completely inhibited HSV-1 replication, but HSV-2 replication was abolished. The drugs appear to be continuously active throughout the infectious cycle. Infectious HSV particles appeared to become inactivated during or soon after egress from the cell. The two triterpenoid drugs lowered the number of virus particles made, and to a much greater extent reduced the infectious virus yield; thus, the progeny virus quality is greatly diminished. HSV-2 infections were more readily inhibited by either CCX or CBX than were HSV-1 infections. PMID- 2991441 TI - Antibody response to varicella-zoster virus surface glycoproteins in chickenpox and shingles. AB - Varicella-zoster virus (VZV)-infected cell surface proteins were investigated using extrinsic radiolabelling of the cell surface, immunoprecipitation of detergent-solubilized extract of the same cell surface and fractionation of the immunoprecipitates using SDS-PAGE. Glycosylated proteins were identified by their affinity for Ricinus communis lectin. Six glycoproteins with apparent mol. wt. of 170K, 105K, 93K, 81K, 53K and 45K were identified. The 170K glycoprotein was shown to be disulphide-linked since under reducing conditions for SDS-PAGE it was cleaved to a protein of 63K mol. wt. The IgG responses to these glycoproteins during various clinical circumstances are described. In acute sera from all chickenpox patients and in the majority of acute shingles sera, antibodies reactive with glycoproteins could not be detected. In chickenpox convalescence, antibodies reactive with glycoproteins of mol. wt. 170K, 105K, 53K and 45K were identified, whilst during zoster convalescence antibodies to all six were prominent. Antibodies to the disulphide-linked glycoprotein persisted for many years following both the primary disease and its reactivation. Disseminated zoster was associated with significantly low levels of antibodies to these surface glycoproteins. PMID- 2991442 TI - Affinity-purified varicella-zoster virus glycoprotein gp1/gp3 stimulates the production of neutralizing antibody. AB - Varicella-zoster virus glycoprotein gp1/gp3 was purified by affinity chromatography using anti-gp1/gp3 monoclonal antibody 19.1 linked to CNBr activated Sepharose CL-4B. Rabbits immunized with purified glycoprotein gp1/gp3 developed mono-specific neutralizing antibody. PMID- 2991443 TI - Production of cell-mediated immune response to herpes simplex virus by immunization with anti-idiotypic heteroantisera. AB - Three BALB/c monoclonal antibodies capable of neutralizing herpes simplex virus type 1 (HSV-1) were used to prepare rabbit anti-idiotypic antisera. Affinity purified antibodies from four of these rabbits were used to immunize mice by repeated subcutaneous injection over a period of 6 to 7 weeks: the mice were then challenged with HSV-1 subcutaneously in the ear pinna. Measurement of ear swelling showed that prior administration of anti-idiotypic serum could generate dose-dependent delayed-type hypersensitivity responses. PMID- 2991444 TI - Susceptibility to herpes simplex virus type 1 infection of non-permissive rat XC(HPRT-) x permissive mouse L(TK-) hybrid cells. AB - Somatic cell hybrids between rat XC(HPRT-) cells, non-permissive for herpes simplex virus type 1 (HSV-1) infection, and permissive mouse L(TK-) cells were constructed and karyotyped. Infection of these hybrid cells by HSV-1 strains F and MP revealed that they were susceptible to the virus. The amounts of virus produced by the hybrid cells, as well as the cytopathic effect observed, was very similar to that of the parental L(TK-) cells. Our results suggest that failure of HSV-1 to replicate in XC cells is more likely to be due to the absence of cellular elements required for efficient virus multiplication rather than to the presence of blocking or inhibiting factors. PMID- 2991445 TI - A comparison of the genomes of bovine herpesvirus type 1 and pseudorabies virus. AB - The DNA sequence homology between bovine herpesvirus type 1 (BHV-1) and pseudorabies virus (PRV) was examined. Reciprocal cross-hybridization of viral DNA labelled by nick translation to Southern blots of restriction endonuclease digested DNA detected homologous sequences dispersed throughout the genomes of the two viruses. The DNA-DNA hybrids formed were stable under high-stringency wash conditions. Sequences of a 32P-labelled PRV DNA fragment probe were found to hybridize only to a specific region of the BHV-1 genome, suggesting that the detected sequence homology was not due to fortuitous hybridization of G + C-rich sequences. As measured by liquid reassociation kinetics the homology between these two viruses was approximately 8%. PMID- 2991446 TI - Variation in the HindIII restriction fragments of DNA from the Chinese Tian Tan strain of vaccinia virus. AB - HindIII fragments D to P of DNA from a Chinese vaccine strain (Tian Tan) of vaccinia virus have been molecularly cloned into the plasmid pAT153 at the unique HindIII site. The Chinese strain DNA differs from a non-vaccine American strain (WR) in having an additional HindIII fragment (P). Twelve HindIII D clones and 12 HindIII F clones of the Chinese strain were analysed by digestion by EcoRI, BamHI, PstI and XhoI. Two forms of D (designated a and b) and of F (a and b) were demonstrated. In each the differences were detected as the presence of an additional EcoRI site, in Da and Fa. The HindIII Fa and Fb fragments of the Chinese strain were shown to differ significantly from the WR strain in their restriction site maps. PMID- 2991447 TI - Molecular cloning of cDNA encoding gp68 of adult T-cell leukaemia-associated antigen: evidence for expression of the pX IV region of human T-cell leukaemia virus. AB - A 2.3 kb cDNA was cloned from human T-cell leukaemia virus [HTLV(MT-2)] virion RNA using a vector system, as plasmid pHTLV 707. The restriction endonuclease map of pHTLV 707 revealed that the insert contained the 5' half of the env gene and a portion of the pX region of HTLV, corresponding to the subgenomic RNA derived from 32S defective HTLV. Nucleotide sequence analysis of pHTLV 707 indicated that the clone contained an open reading frame for a 60K mol. wt. protein including the upstream and entire pX IV region. A rabbit antibody raised against a synthetic decapeptide deduced from the nucleotide sequence at the carboxyl terminus of the pX IV region immunoprecipitated gp68, and also 80K and 40K proteins. PMID- 2991448 TI - Antibody-dependent enhancement of tick-borne encephalitis virus infectivity. AB - Fourteen mouse monoclonal antibodies raised against tick-borne encephalitis virus (TBEV) and polyclonal antisera raised against six other flaviviruses, Edge Hill (EHV), Japanese encephalitis (JEV), Langat (LGTV), louping ill (LIV), West Nile (WNV) and yellow fever (YFV), were tested for their ability to enhance the replication of TBEV in cells of the mouse macrophage-like line P388 D1, and for their reactivity in ELISA and haemagglutination inhibition (HI) tests. Irrespective of their specificity for either the 51K or 58K polypeptide present in TBEV-infected cells, 13 of the 14 monoclonal antibodies enhanced the replication of TBEV but not of WNV. The remaining monoclonal antibody, which immunoprecipitated the 58K polypeptide of TBEV enhanced WNV but not TBEV, although it reacted strongly with both viruses in ELISA and HI tests. Only polyclonal antisera against viruses within the tick-borne encephalitis virus complex (TBEV, LGTV and LIV) enhanced TBEV replication, although all the polyclonal antisera reacted with TBEV by ELISA; two (against JEV and WNV) also reacted by HI test and all enhanced the replication of WNV. These findings suggest that with TBEV, enhancement may be TBEV complex-specific rather than flavivirus-specific. Data derived from testing both polyclonal and monoclonal antibodies suggest further that not all antibodies that bind to the envelope glycoprotein of TBEV are able to enhance the replication of TBEV, and that enhancement is epitope-specific. PMID- 2991449 TI - Differential coupling of GABA-A and GABA-B receptors to the noradrenergic system. AB - The GABA-A receptor agonist THIP, or the mixed GABA-A/GABA-B receptor agonist progabide dose dependently increased the release of norepinephrine (as measured by the production of MHPG) in the cerebral cortex and hippocampus. This effect was partially reversed by treatment with the GABA-A receptor antagonist bicuculline. In contrast, the GABA-B receptor agonist baclofen decreased the release of norepinephrine in the cerebral cortex and hippocampus. Pretreatment with the presynaptic noradrenergic neurotoxin DSP4 increased the Bmax for beta adrenergic receptor binding in the cerebral cortex and hippocampus. This effect was partially prevented by chronic (14 day) treatment with either the beta adrenergic agonist clenbuterol or the GABA-B receptor agonist baclofen. In contrast, chronic (14 day) administration with either the GABA-A receptor agonist THIP or the antidepressant imipramine failed to alter the increase in beta adrenergic receptor binding produced by DSP4 pretreatment. These data suggest that the GABA-A receptor may be coupled to the presynaptic noradrenergic neuron and modulate the release of norepinephrine, while the GABA-B receptor is coupled to the postsynaptic noradrenergic neuron and likely functions through the cyclic AMP generating system. PMID- 2991450 TI - Cytochrome c oxidase: structure, function, and physiological role. Rome and Caprarola, Italy, 3-6 October 1984. PMID- 2991451 TI - The protein formula of beef heart cytochrome c oxidase. AB - Beef heart cytochrome c oxidase consist of 12 different polypeptides stoichiometrically arranged in respiratory complex IV. The functional 2 heme a, 2 copper monomer of this complex consist of 1793 amino acids; the exact Mr is 202,787 Da. From 17 cysteine residues, six are involved in the formation of three disulphide bonds. The theoretical heme a content of the enzyme is 9.86 nmol/mg protein. The theoretical iron and copper contents are 0.55 and 0.63 microgram/mg protein, respectively. PMID- 2991452 TI - Cytochrome oxidase: structural insights from electron microscopy and from secondary structure prediction. AB - Electron microscopic images of selectively contrasted cytochrome oxidase dimer crystals are interpreted in a manner consistent with the structure of monomers determined by Fuller et al. (J. Molec. Biol. 134, 305-327). The arms of the y shaped monomers lie within and perpendicular to the lipid bilayer protruding approximately 25 A on the matrix side of the membrane. The cytoplasmic-side tails of two monomers spread apart in a dimer forming a large cleft. Decoration of the exposed matrix side of vesicle crystals with antisubunit IV antibody fragments indicates that subunit IV lies along the a-crystal axis roughly 20 A from the center of the dimer. A membrane propensity algorithm applied to the sequences of cytochrome oxidase subunits predicts a total of 19 transmembrane alpha-helices per monomer. PMID- 2991453 TI - Three-dimensional reconstruction of cytochrome oxidase vesicle crystals prepared by cholate solubilization. AB - Cytochrome oxidase vesicle crystals with long-range order have been obtained from cholate-solubilized, highly purified reconstitutively active preparations. These crystals, which are suitable for electron-microscopic structure investigation, show pgg symmetry in the 0 degree projection. Using Fourier reconstruction and modified back-projection methods, a three-dimensional reconstruction has been obtained at a resolution of 25 A. Our structural results are in agreement with the model of Henderson et al. [J. Mol. Biol. 112, 631 (1977)] obtained for their Triton-derived crystals. PMID- 2991454 TI - Diphosphatidylglycerol reactivation of lipid-depleted beef heart cytochrome c oxidase: effect of subunit III removal. AB - Beef heart cytochrome c oxidase can be fully delipidated by Triton X-100 using an alkaline pH, high ionic strength incubation followed by glycerol gradient centrifugation at pH 8 in 1% Triton X-100. Unfortunately, this procedure removes not only the 2-3 diphosphatidylglycerol (DPG) molecules that are tightly bound to each heme aa3 complex, but also removes subunit III. A one-to-one correlation exists between the percent of subunit III removed and the percent of tightly bound DPG extracted, suggesting a possible tight binding of these two molecules. However, based upon regeneration of most of the electron transport activity of the subunit III deficient complex by exogenous DPG, it appears that the functional DPG binding site must be located on a subunit other than subunit III. PMID- 2991455 TI - Helical packing in the hydrophobic sector of cytochrome c oxidase. AB - An arrangement for the membrane-spanning segments of the three larger subunits of cytochrome c oxidase is proposed on the basis of sequence comparison and polarity distribution estimated from the data available for 11 different organisms. PMID- 2991456 TI - Cloning of Paracoccus cytochrome c oxidase subunit II. AB - Cytochrome c oxidase from Paracoccus denitrificans is composed of two subunits, yet is active in both electron transport and proton translocation. A cloning approach and immunologic screening protocol is described for the isolation of the subunit II gene expressed in E. coli. DNA sequencing should establish the extent of homology to eukaryotic oxidase. PMID- 2991457 TI - The structure of the cytochrome a3-CuB site of mammalian cytochrome c oxidase as probed by MCD and EPR spectroscopy. AB - The nature of the complexes formed between cytochrome c oxidase and the three inhibitory ligands N3-, CN-, and S2- have been investigated by a combination of MCD and EPR spectroscopy. CN- forms a linear bridge between the Fe III a3 and CuB II, suggesting that the distance between these centers in the oxidized enzyme is between 5 and 5.25 A. This distance is too short to permit N3- to form a linear bridge and the evidence suggests this to be bent. In contrast S2- or SH- is unable to form any bridge and it seems likely that two SH- ions are bound by the bimetallic site, one to Fe III a3 and the other to CuB I. The significance of the a3-CuB distance in terms of oxygen binding and reduction is discussed. PMID- 2991458 TI - Effect of cyanide binding on the copper sites of cytochrome c oxidase: an X-ray absorption spectroscopic study. AB - Cu x-ray absorption spectroscopy (XAS) has been used to investigate the effect of cyanide treatment on the structures of the copper sites in beef heart cytochrome c oxidase. The Cu K-edge spectrum changes significantly upon cyanide binding to resting state enzyme, as does the Cu extended x-ray absorption fine structure (EXAFS) spectrum. The Cu EXAFS Fourier transfer (FT) exhibits an enhanced peak for the cyanide-treated enzyme in the region containing the Cu...Fe peak in the resting state FT (at R' approximately equal to 2.6-2.7 A). This peak in the cyanide-treated sample is hypothesized to arise from "outer shell" scattering from a linear Cu-cyanide moiety, suggesting cyanide binding to CuB only (CuB 2+ CN-) or cyanide bridging between the Fe of heme a3 and CuB (Fe3+-(CN-)-CuB 2+). PMID- 2991460 TI - The influence of electrostatic interactions between buried charges on the properties of membrane proteins. AB - The importance of electrostatic interactions between buried charges in determining the properties of membrane proteins is considered. It is demonstrated that in some cases altered properties may result from the extraction of a membrane protein into an aqueous medium even when the protein conformation is unperturbed. PMID- 2991459 TI - Multiple structures and functions of cytochrome oxidase. AB - X-ray absorption studies have been used to investigate the structure of the four redox centers (2Fe, 2Cu) of the terminal enzyme in the respiratory chain, cytochrome c oxidase in the resting oxidized form as well as in the functional intermediates that are freeze-trapped. Methods of x-ray fluorescence detection for these low-concentration samples together with low-temperature cryostats and simultaneous optical monitoring were developed to ensure good signal-to-noise data and sample integrity. The resting oxidized form contains a sulfur bridge between the copper and iron of the active site which are separated by approximately 3.8 A. This separation of the active site metal atoms was uniquely identified by comparison of both the iron and copper EXAFS data and iron EXAFS of the copper-depleted enzyme. In the reduced state, the CO or O2 is bound to the active site iron having a structure identical to CO or oxy hemoglobin while the sulfur remains with the active site copper. Little change in structure is observed for the other iron and copper. It is the sulfur bridged active site form that is isolated by the Yonetani and Caughy methods with greater than or equal to 85% homogeneity but not the Hartzell-Beinert or similar methods. Another form observed in the redox cycle is also fully oxidized but lacks the sulfur bridged active site with the iron of the active site having a structure identical to that of the peroxidases. This form exhibits peroxidase as well as oxidase activity, and a stable intermediate is formed with hydrogen and ethylhydrogen peroxide in which the iron of the active site is structurally similar to that of the peroxidase intermediate. The active site copper, however, does not participate in the peroxidatic role and the structures of the other iron and copper are identical to those of the sulfur bridged resting oxidized form. Thus this unique enzyme has peroxidase activity which may serve to safeguard its main oxidase function. PMID- 2991461 TI - The photoreactivity of the copper-NO complexes in cytochrome c oxidase and in other copper-containing proteins. AB - The complexes of NO with CuB of cytochrome c oxidase in which cytochrome a3 may or may not be ligated to cyanide or fluoride are photodissociable. NO does not appear to react with CuB in complexes of cytochrome c oxidase in which sulphide or mercaptans are ligated to the haem iron of cytochrome a3. A comparison is made between the photoreactivity of the complexes of NO with cytochrome c oxidase and those with ceruloplasmin, ascorbate oxidase, and haemocyanin. It is shown that the photoreactivity of CuB 2+.NO in cytochrome c oxidase is not unique for this enzyme, but may also be observed in the complexes of NO with type-1 copper containing enzymes. This would suggest that the ligation of CuB in cytochrome c oxidase shows some similarity to type-1 copper in blue oxidases. PMID- 2991462 TI - The steady-state rate equation for cytochrome c oxidase based on a minimal kinetic scheme. AB - A minimal catalytic cycle for cytochrome c oxidase has been suggested, and the steady-state kinetic equation for this mechanism has been derived. This equation has been used to simulate experimental data for the pH dependence of the steady state kinetic parameters, kcat and Km. In the simulations the rate constants for binding and dissociation of cytochrome c and for two internal electron-transfer steps have been allowed to vary, whereas fixed experimental values (for pH 7.4) have been used for the other rate constants. The results show that the dissociation of the product, ferricytochrome c, cannot be rate-limiting under all conditions, but that intramolecular electron-transfer steps also limit the rate. They also demonstrate that Km can differ considerably from the dissociation constant for the cytochrome c-oxidase complex. Published values for the rate constant for the dissociation of ferricytochrome c are too small to account for the steady-state rates. It is suggested that, at high concentrations, ferryocytochrome c transfers an electron to a cytochrome c molecule which remains bound to the oxidase. This can also explain the nonhyperbolic kinetics, which is observed at low substrate concentrations. PMID- 2991463 TI - Flow-flash, time-resolved resonance Raman spectroscopy of the oxidation of reduced and of mixed valence cytochrome oxidase by dioxygen. AB - Time-resolved resonance Raman spectroscopy has been used to study the reduction of oxygen by both reduced and mixed valence cytochrome oxidase. Laser flash photodissociation of CO from the carbon monoxy complex of the enzyme, after this species had been rapidly mixed with oxygenated buffer, was used to initiate the reaction for both forms of the enzyme. The CO photolysis product of the mixed valence enzyme contains cytochrome a3+ and cytochrome a3(2+) in its unligated form. This species reacts with O2 in the first few microseconds to form a photolabile intermediate which has Raman frequencies characteristic of oxygenated heme. This indicates that an oxyhemoglobinlike complex of oxygen with a3(2+) is the precursor to oxygen reduction. A similar intermediate is detected in the fully reduced enzyme reaction. In the mixed valence oxidase system, the oxy intermediate is replaced by a nonphotolabile species in which a3 is oxidized with t1/2 approximately equal to 200 musec. These results demonstrate the feasibility of applying time-resolved vibrational techniques to irreversible electron transfer reactions and, in particular, elucidate some of the transient species in the cytochrome oxidase/O2 system. PMID- 2991465 TI - Evidence for a photoactivation of CO rebinding to fully reduced cytochrome c oxidase after low-temperature flash photolysis. AB - Carbon monoxide rebinding to isolated fully reduced cytochrome c oxidase has been investigated by low-temperature, flash photolysis, dual-wavelength spectrometry. By using separately different wavelength pairs to monitor the liganding of CO to Fe a3 and by keeping all other experimental conditions identical, there has been singled out a photoactivation effect on CO rebinding. For instance, at 187 K, the rate constant of CO rebinding observed at 425-475 nm was twice that derived from the kinetic at 444-475 nm despite a rate constant of photodissociation about 10 times larger at 425-475 nm than at 444-475 nm. This new finding is discussed with respect to previous investigations under similar conditions. PMID- 2991464 TI - A stopped-flow study of the reaction of reduced cytochrome oxidase with oxygen. AB - The reaction of a reduced cytochrome oxidase system consisting of beef heart cytochrome oxidase, cytochrome c, and ascorbate with molecular oxygen was kinetically and thermodynamically investigated using a stopped-flow, rapid wavelength-scanning technique. Processes for oxidation of ferrocytochrome a, bound ferrocytochrome c, and free ferrocytochrome c have been identified, and their rate constants have been determined. Values of the activation energy for these reactions indicate that the oxidation of bound ferrocytochrome c is a simple chemical electron-transfer process and that oxidations of ferrocytochrome a and free ferrocytochrome c are complex processes involving changes in protein conformation. PMID- 2991466 TI - Exclusive CO binding to cytochrome oxidase. AB - CO added to dithionite-reduced cytochrome oxidase pretreated with azide, cyanide, or fluoride yielded CO-ferrous heme a3 trapping the unliganded reduced heme. Ferrous heme a3 was either an equilibrium species initially present, or provided by dissociation of ligand-bound ferric heme a3 followed by the reduction with dithionite. In the latter case the ligand dissociation was rate-limiting for the CO compound formation. Pretreatment of the enzyme with the inhibitory ligands affected neither photodissociation and reassociation of the CO compound thus formed, nor reaction with dioxygen initiated by the flow-flash method to any significant degree. Only the cyanide treatment slightly decreased the rate of intramolecular electron transfer. These results indicate that no inhibitory ligand but CO remains in the vicinity of the heme a3-CuB center in the CO compound of cytochrome oxidase. PMID- 2991467 TI - A comparative survey of several bacterial aa3-type cytochrome c oxidases. AB - The aa3-type cytochrome c oxidases purified from Nitrobacter agilis, Thiobacillus novellus, Nitrosomonas europaea, and Pseudomonas AM 1 were compared. They have haem a and copper atom as the prosthertic groups and show alpha and gamma absorption peaks at around 600 and 440 nm, respectively. Each oxidase molecule is composed of two kinds of subunits. The N. agilis oxidase has 2 moles of haem a and 2 atoms of copper in the minimal structural unit composed of one molecule each of the two kinds of subunits, while the T. novellus enzyme seems to contain one molecule of the haem and one atom of the metal in the unit. The N. europaea oxidase shows very low affinity for carbon monoxide. Each oxidase reacts rapidly with some eukaryotic cytochromes c as well as with its native cytochrome c. The cytochrome c oxidase activity of the N. agilis oxidase is 50% inhibited by 1 microM KCN, while 50% inhibition of the activity requires 100 microM KCN in the case of the N. europaea enzyme. PMID- 2991468 TI - Potentiometric study of cytochrome c1aa3 from Thermus thermophilus. AB - We have examined the redox behavior of the cytochrome c1aa3 complex from Thermus thermophilus. In potentiometric titrations the cytochrome c behaves as an independent center having n = 1 and E = 205 mV (NHE). Under the assumption that the individual centers equilibrate independently in this experiment, changes in the absorption band at 603 nm have been resolved into two components: cytochrome a (n = 1, Em = 270 mV, 60% spectral contribution) and cytochrome a3 (n = 2, Em = 360 mV, 40% spectral contribution). The n = 2 process was attributed to strong chemical coupling between cytochrome a3 and CuB. The enzyme was also titrated with a mixture of NADH and PMS, and the results are shown not to conform to a model of intramolecular equilibrium according to the equilibrium constants obtained from the potentiometric titration. It is suggested that a conformational equilibrium within the complex may control electron transfer between cytochromes a and a3. PMID- 2991470 TI - The reactivity of pulsed cytochrome c oxidase toward carbon monoxide. AB - When pulsed cytochrome c oxidase is exposed to carbon monoxide in the absence of oxygen the enzyme is converted quickly to its CO-associated mixed valence state. The half-time for this reaction at 0 degree C is about 4 min. This is about 100 times faster than a similar reaction which begins with the resting form of the enzyme. The possible significance of this reaction in understanding the pulsed/resting phenomenon and the carbon monoxide oxygenase reactions of cytochrome oxidase is discussed. PMID- 2991469 TI - Pulsed cytochrome c oxidase. AB - The identification of two functionally distinct states, called pulsed and resting, has led to a number of investigations on the conformational variants of the enzyme. However, the catalytic properties of cytochrome oxidase may depend on a number of experimental conditions related to the solvent as well as to the protocol followed to determine the turnover number of the enzyme. This paper reports results which illustrate that the steady-state differences between pulsed and resting oxidase may, or may not, be detected depending on experimental conditions. PMID- 2991471 TI - Quantization of membrane potential generation by cytochrome c oxidase in small vesicles. AB - Proteoliposomes incorporating cytochrome c oxidase have been prepared by the cholate dialysis method and by sonication. Sonication produces multilamellar vesicles heterogeneous in size in contrast to a more uniform preparation of unilamellar vesicles produced by the dialysis procedure. Respiratory control in both preparations ranges between 4 and 8. From an electron microscopic analysis of proteoliposome size, the average electrical capacitance/vesicle for the dialyzed and sonicated preparations is calculated as 15 X 10(-18) F and 130 X 10( 18) F, respectively. These capacitance values would lead to a quantization of membrane potential generation by the enzyme at 77 mV/turnover for the dialyzed preparation and 9 mV/turnover for sonicated vesicles. It is argued that these differences can explain the dependence of H+ translocation on the number of turnovers of cytochrome c oxidase in dialyzed preparations in contrast to the lack of dependence on number of turnovers in sonicated preparations. PMID- 2991472 TI - Separation of enzymically active bovine cytochrome c oxidase monomers and dimers by high performance liquid chromatography. AB - The aggregation state of two types of bovine heart cytochrome c oxidase preparations in the presence of laurylmaltoside was investigated by high performance liquid chromatography in two buffers of ionic strengths of 388 mM and 45 mM, respectively. At high ionic strength, it was found that the Fowler cytochrome c oxidase preparation was monomeric (Mr = 2 X 10(5)), while monomers and dimers (2 X aa3, Mr = 4 X 10(5)) could be isolated from the Yonetani preparation. Under these conditions there was no rapid equilibrium between the two forms. Covalent cytochrome c oxidase-cytochrome c complexes were largely dimeric, and addition of ascorbate and cytochrome c to the oxidase also promoted dimerization. At low ionic strength (I = 45 mM) in the presence of laurylmaltoside the oxidase and the covalent complex with cytochrome c were largely monomeric. In the steady-state oxidation of ferrous horse heart cytochrome c, the monomeric enzyme displayed biphasic kinetics at I = 45 mM. This suggests that the presence of high- and low-affinity reactions is an intrinsic property of the cytochrome c oxidase monomer. PMID- 2991473 TI - The optimal technological approach to the development of human hybridomas. PMID- 2991474 TI - Ethylene oxide polyneuropathy: clinical follow-up study with morphometric and electron microscopic findings in a sural nerve biopsy. AB - A case is reported of ethylene oxide polyneuropathy after 5 months of exposure. There was symmetrical distal weakness of both lower extremities and transitory reduced nerve conduction velocities with increased latencies. Sural nerve biopsy revealed nerve fibre degeneration of the Wallerian type, associated with reduction of axonal cross-sectional areas and some degree of nerve fibre regeneration that could be confirmed morphometrically. In addition, there was conspicuous paranodal vesicular disintegration of individual myelin lamellae. Unusual cisternae with introverted hemidesmosomes were noted in endoneurial fibroblasts. PMID- 2991475 TI - Effects of intravenous hyperalimentation during treatment in patients with small cell lung cancer. AB - One hundred nineteen patients were entered onto a randomized trial of the role of intravenous hyperalimentation (IVH) in patients with small-cell lung cancer. IVH was given during the first 30 days of induction chemotherapy to 54 patients. IVH did not effect any improvement in response or survival from therapy. In view of the lack of benefits from IVH, an analysis was made of the toxicities suffered by the 54 patients receiving IVH as well as any effects IVH might have made on chemotherapy-induced toxicity. Toxicities observed included mechanical difficulties with the catheter leading to temporary or permanent discontinuation of the IVH (11 patients), subclavian vein thrombosis (one patient), sepsis in nine patients v none of the 62 control patients, fluid overload (27 patients), hyponatremia (25 patients), and hyperglycemia requiring insulin (13 patients). Patients receiving IVH had higher granulocyte counts on days 14 and 21 of the first cycle of chemotherapy. Analysis shows that this difference is likely caused by fever and infection associated with IVH rather than any nutritional effect on granulopoiesis. In this population of patients, IVH had significant complications but did not ameliorate chemotherapy-induced toxicity and it did not effect any clinical benefit. Future studies of adjunctive nutritional therapy must consider the significant risk in this older population and must limit IVH volume or exclude patients with even mild compromise in cardiovascular functions. Further, any new trial must have a significant rationale for adjunctive use to justify the potential risks. PMID- 2991476 TI - High-dose cyclophosphamide in small-cell carcinoma of the lung. AB - Eighteen patients with untreated small-cell cancer of the lung have been treated with cyclophosphamide 200 mg/kg on two occasions at an interval of four weeks. An additional eight patients received etoposide in addition to cyclophosphamide. Measurements of tumor volume were made by thoracic computed tomographic (CT) scan before and after each cycle. When compared with our previous study in which only one cycle of cyclophosphamide was given, the double procedure did not increase response rate, decrease the incidence of local relapse, or prolong the relapse free interval. Survival after relapse was shorter with two cycles of chemotherapy mainly due to greater difficulty in administering further chemotherapy. The CT scans showed a decrease in tumor volume from 99.2 cc to 21 cc after the first cycle, but a smaller decrease to 14.1 cc after the second. These results show that the rapid emergence of drug resistance imposes limitations on the use of very high-dose cytotoxic chemotherapy. PMID- 2991477 TI - Effect of initial resection of small-cell carcinoma of the lung: a review of Southwest Oncology Group Study 7628. AB - The role of surgery in small-cell carcinoma of the lung (SCCL) has been recently re-evaluated. We reviewed the records of 262 patients with limited SCCL on Southwest Oncology Group (SWOG) protocol 7628. Fifteen patients were identified who presented after surgical resection (12 lobectomy, three pneumonectomy). All patients were subsequently treated with chemotherapy, radiotherapy +/- immunotherapy (BCG). Median survival time was 10.5 months. Median survival time of patients with initial surgical resection was 25 months (P = .004). Forty-five percent of the surgical patients were alive at two years v 13.7% of the nonsurgical patients (P less than .05). A second subgroup of 33 patients was identified with small primary tumors who did not undergo surgical resection. Median survival time in this group was ten months (P = .03). Site of initial relapse was clearly documented in 142 patients. Fifty-six percent of patients not receiving surgery had initial relapse within the chest compared to 13% of patients undergoing surgery (P = .002). Whether the survival benefit identified was caused by or was incidental to surgical resection of the primary lesion remains to be determined in randomized prospective trials of operable candidates. PMID- 2991478 TI - Combined modality therapy with radiotherapy, chemotherapy, and immunotherapy in limited small-cell carcinoma of the lung: a Phase III cancer and Leukemia Group B Study. AB - Patients with limited-stage small-cell carcinoma of the lung (SCCL) were randomly assigned to a four-drug chemotherapy program consisting of methotrexate, doxorubicin, cyclophosphamide, and CCNU (MACC) or to a regimen consisting of cyclophosphamide, CCNU, and vincristine alternated with Adriamycin (Adria Laboratories, Columbus, Ohio) and vincristine (CCV/AV). All patients received 4,500 cGy, in a split course, to the primary tumor, mediastinum, and supraclavicular lymph node drainage areas and 3,000 cGy to the whole brain. After four cycles of chemotherapy, patients were randomly assigned to chemotherapy plus methanol extractable residue of BCG (MER-BCG) or no MER-BCG. The complete response frequencies were similar for the two regimens (54% and 48%) as were the median survivals (12.0 and 11.5 months) and the two-year survival rates (15% and 17%). Immunotherapy with MER-BCG did not prolong the time to disease progression or improve survival. Women had a greater chance of achieving a complete remission independent of performance status. There was a complex interaction between sex and the chemotherapy regimens that may have important implications for the design and stratification of future trials in SCCL. PMID- 2991479 TI - Glutamic acid decarboxylase immunoreactivity in layer IV of barrel cortex of rat and mouse. AB - The morphology and distribution of neurons and terminals that are immunoreactive to glutamic acid decarboxylase (GAD) were investigated in barrel cortex of the rat and mouse. The morphology of the GAD-immunoreactive neurons located in layer IV of the barrel field resembles that of the large, smooth stellate neurons described previously in Golgi studies. Most of the somas of GAD-positive neurons are located along the sides of the barrels. They constitute about 13 to 15% of the total neuronal population in layer IV. The spatial distribution of GAD positive terminals in layer IV is similar to the distribution of GAD-positive somas. Very few GAD-positive neurons and terminals are found in the septal regions. This unique distribution of GAD immunoreactivity in the barrel cortex may serve as a model to study cortical inhibitory mechanisms. PMID- 2991480 TI - Physiological and morphological analysis of synaptic transmission between leech motor neurons. AB - In the leech Hirudo medicinalis inhibitory motor neurons to longitudinal muscles make central inhibitory connections with excitatory motor neurons to the same muscles. We have used a variety of physiological and morphological methods to characterize these inhibitory connections. The efficacy of the transmission between the inhibitors and the excitors was measured by using three intracellular electrodes, two in the inhibitor (one for injecting current and one for measuring voltage) and a third electrode in the excitor for measuring the resultant voltage changes. We have determined that delta Vpre/delta Vpost, or what we have called the transmission coefficient, is X = 0.51, as measured in the somata of the two cells. Evidence which we have obtained leads us to propose that these inhibitory connections between motor neurons are probably monosynaptic. The synaptic latency is consistent with a monosynaptic connection. In addition, a double-labeling technique, whereby one neuron was filled with Lucifer Yellow and the other with horseradish peroxidase (HRP), was used to determine the anatomical relationship between inhibitors and excitors in whole mounts. This revealed varicosities on the processes of inhibitor motor neurons which appear to make contact with processes of excitor motor neurons. A second double-labeling technique, whereby one neuron was filled with HRP and the other with an electron-dense particulate marker, revealed adjacent processes between an inhibitor and an excitor in electron microscopic thin sections which could be the sites of synaptic contact between the neurons. The connections appear to be mediated largely by graded transmitter release from the inhibitory motor neurons. Three different methods were used to demonstrate that synaptic transmission remained in the absence of impulses in the inhibitory motor neurons. These included eliminating the impulse supporting portion of the motor neuron by pinching off its axon, abolishing impulses by replacing Na+ with Tris in the medium bathing the nerve cord, and increasing the threshold for impulse production by raising the Mg2+ and Ca2+ concentrations in the medium bathing the nerve cord. PMID- 2991481 TI - Nonsynaptic characteristics of neurotransmission mediated by egg-laying hormone in the abdominal ganglion of Aplysia. AB - The bag cell neurons of the marine mollusk Aplysia are a putative multitransmitter system which utilizes two or more neuropeptides that are enzymatically cleaved from a common precursor protein. It has been proposed that one of the neuropeptides, egg-laying hormone (ELH), acts nonsynaptically as a neurotransmitter in the abdominal ganglion by diffusing long distances to target neurons compared to conventional transmitters acting at synapses. To test this idea further, we investigated the physiological properties of neurotransmission mediated by ELH. We found that ELH acts directly to duplicate two types of responses produced by a burst discharge of the bag cells: prolonged excitation of LB and LC cells, and the previously described effect of ELH, burst augmentation of cell R15. Analysis of perfusate collected after electrical stimulation of the bag cells showed that the peptide is released in sufficient quantity to diffuse long distances within the ganglion without being completely inactivated. To mimic the way the peptide is thought to be released physiologically, ELH was arterially perfused into the ganglion. The response normally produced by bag cell activity was duplicated by 0.5 to 1.0 microM concentrations of ELH and showed no rapid desensitization. ELH had no effect on cells that are unaffected by bag cell activity and no effect on cells that are inhibited (LUQ cells) or transiently excited (cells L1 and R1) by bag cell activity. Acidic peptide, another peptide encoded on the ELH precursor protein, was found to be synthesized and released by the bag cells, but it had no effect on the cells we tested. We conclude that the combined properties of ELH neurotransmission resemble the properties of transmission at autonomic nerve endings on cardiac and smooth muscle rather than those of conventional synaptic transmission. ELH released from bag cells is dispersed throughout the interstitial and vascular spaces of the ganglion to produce responses in the cells that have receptors for the peptide. The results also suggest that ELH mediates only a subset of the responses induced by bag cell activity; they are consistent with data indicating that the other responses are mediated by other bag cell peptides derived from the same precursor protein as ELH. PMID- 2991482 TI - Specific labeling of rat brain substance P receptor with [3H]physalaemin. AB - The binding of [3H]physalaemin [( 3H]PHY) to rat brain membranes is specific, saturable and reversible in the presence of monovalent cations and peptidase inhibitors. Monovalent cations increase the binding of [3H]PHY in an ionic strength (mu)-dependent manner with an optimal effect at mu higher than 0.3. Addition of 2.5 mM MnCl2 results in a 2-fold increase in the affinity (KD) and a 40% increase in the maximal receptor density (Bmax). Scatchard analysis under these conditions indicates the existence of a single population of noninteracting sites with KD of 3.6 nM and a Bmax of 76 fmol/mg of protein. Substance P (SP) and physalaemin are equipotent in inhibiting the binding of [3H]PHY, whereas the potency of SP(2-11), SP(3-11), and SP(4-11) decreased in inverse proportion to their length. The relative affinity of the different tachykinins, SP, and SP fragments in competing with [3H]PHY correlates with their potency to stimulate several bioassay systems, indicating that [3H]PHY labels a physiologically relevant binding site that correspond to the SP-P tachykinin receptor. Guanine nucleotides completely abolish the increase in the binding of [3H]PHY produced by 2.5 mM MnCl2, but in its absence, the nucleotides reduce binding only by 15%. Guanine nucleotides reduce binding to the same level regardless of the presence or absence of the divalent cation. Regional distribution studies confirm that the density of SP receptors is maximal in the olfactory bulb, followed by the hypothalamus, striatum, hippocampus, cortex, and cerebellum. PMID- 2991483 TI - The ultrastructural basis for synaptic transmission between primary muscle afferents and neurons in Clarke's column of the cat. AB - The synaptic connection between primary muscle afferents and dorsal spinocerebellar tract (DSCT) neurons has been studied in an attempt to reveal some of the mechanisms underlying excitatory transmission in the mammalian central nervous system. Previous electrophysiological experiments have shown that the excitatory postsynaptic potentials (EPSPs) evoked DSCT neurons by impulses in a single muscle afferent fluctuate in amplitude. These fluctuations occur between discrete amplitudes which are separated by quantal increments. Two alternative hypotheses relate such a quantal increment to all-or-nothing transmitter release from either (1) an entire synaptic bouton or (2) an individual transmitter release site, given that a bouton may contain multiple release sites. The present study was undertaken primarily to gain ultrastructural evidence on these proposals. Electrodes filled with horseradish peroxidase (HRP) were used to label single identified group Ia afferent fibers and DSCT neurons in the lumbar spinal cord of anesthetized cats. HRP-labeled Ia synaptic boutons, and the contacts formed between HRP-labeled Ia boutons and the dendrites of a DSCT neuron labeled intracellularly with HRP, were examined in serial sections under the electron microscope. Group Ia boutons were found to contain multiple synaptic specializations, as evidenced by pre- and postsynaptic thickenings and presynaptic clusters of vesicles. Careful examination of a bouton in serial sections revealed each specialization as a separate structure. These observations support the proposal that synaptic transmission between group I muscle afferents and DSCT neurons occurs with discrete all-or-nothing EPSPs associated with transmitter release sites, rather than boutons per se. PMID- 2991484 TI - Biochemical characterization of a filtered synaptoneurosome preparation from guinea pig cerebral cortex: cyclic adenosine 3':5'-monophosphate-generating systems, receptors, and enzymes. AB - A particulate preparation was obtained by low speed centrifugation of guinea pig cerebral cortical homogenates prepared with a Krebs-Henseleit buffer. Light microscopic examination, using a reflected light differential interference contrast system, reveals the presence of intact neurons, axonal fragments, glial cells, and erythrocytes along with an abundance of small spherical entities (diameter about 1.1 micron) and snowman-shaped entities (diameter of larger sphere about 1.1 micron, diameter of attached smaller sphere about 0.6 micron). Many unattached smaller spherical entities are also present (diameter about 0.6 micron). Pressure filtration through 5- or 10-micron Millipore filters, followed by low speed centrifugation and resuspension, removes most of the larger entities to afford a suspension composed mainly of the small spherical and snowman-shaped entities. Electron microscopic examination reveals the presence of many synaptosomes with attached resealed postsynaptic entities. It is proposed that these correspond to the snowman-shaped entities to be termed synaptoneurosomes. Accumulations of cyclic AMP elicited by 2-chloroadenosine and histamine, and by combinations of 2-chloroadenosine, histamine, norepinephrine, and forskolin, are lower in filtered than in unfiltered preparations, whereas accumulations elicited by forskolin are unchanged. Levels of adenylate cyclase are reduced by filtration, whereas levels of phosphodiesterase are unchanged. Filtration reduces levels of markers for whole cells and endothelial cells, whereas neuronal markers such as acetylcholinesterase activity and norepinephrine uptake are increased. Levels of S-100 protein, a marker for glial cells, are not significantly decreased. There is no apparent change in the density of many receptors or ion channels. Levels of A1-adenosine and H1-histamine receptors are increased, whereas levels of so-called peripheral benzodiazepine-binding sites are decreased. PMID- 2991485 TI - Intracranial germ-cell tumors: natural history and pathogenesis. AB - The natural history of primary intracranial germ-cell tumors (GCT's) is defined from 389 previously published cases, of which 65% were germinomas, 18% teratomas, 5% embryonal carcinomas, 7% endodermal sinus tumors, and 5% choriocarcinomas. Intracranial GCT's display specificity in site of origin. Ninety-five percent arise along the midline from the suprasellar cistern (37%) to the pineal gland (48%), and an additional 6% involve both sites. The majority of germinomas (57%) arise in the suprasellar cistern, while most nongerminomatous GCT's (68%) preferentially involve the pineal gland (p less than 0.0001). The age distribution of afflicted patients is unimodal, centering with an abrupt surge in frequency in the early pubertal years; 68% of patients are diagnosed between 10 and 21 years of age. Nongerminomatous GCT's demonstrate an earlier age of onset than do germinomas (p less than 0.0001). Prolonged symptomatic intervals prior to diagnosis are common in germinomas (p = 0.0007), in suprasellar GCT's (p = 0.001), and among females (p = 0.02). Parasellar germinomas commonly present with diabetes insipidus, visual field defects, and hypothalamic-pituitary failure. Nongerminomatous GCT's present as posterior third ventricular masses with hydrocephalus and midbrain compression. Germ-cell tumors may infiltrate the hypothalamus (11%), or disseminate to involve the third ventricle (22%) and spinal cord (10%). Among a subpopulation of 263 conventionally treated patients, two factors were of prognostic significance: 1) histological diagnosis; germinomas were associated with significantly longer survival than nongerminomatous GCT's (p less than 0.0001); and 2) staging of the extent of disease; this emphasizes the ominous character of involvement of the hypothalamus (p = 0.0002), third ventricle (p = 0.02), or spinal cord (p = 0.01). Specific recommendations regarding the necessity of histological diagnosis and staging of the extent of disease are made in light of modern chemotherapeutic advances. The pathogenesis of GCT's may be revealed by their specificity of origin within the positive (suprasellar cistern-suprachiasmatic nucleus) and negative (pineal) regulatory centers for gonadotropin secretion within the diencephalon. The abrupt rise in age distribution at 10 to 12 years suggests that the neuroendocrine events of puberty are an "activating" influence in the malignant expression of these embryonal tumors. PMID- 2991486 TI - Phase III comparison of BCNU and the combination of procarbazine, CCNU, and vincristine administered after radiotherapy with hydroxyurea for malignant gliomas. AB - The authors report the results of a randomized study conducted to evaluate the relative benefit of treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) or the combination of procarbazine, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, and vincristine (PCV) administered after radiation therapy with hydroxyurea to 76 evaluable patients with glioblastoma multiforme and 72 patients with other anaplastic gliomas. The primary end-point of the study was time to tumor progression. For better-risk patients with Karnofsky performance scores of 70 to 100, results suggest that PCV was of greater benefit than BCNU (p = 0.15 for glioblastoma multiforme; p = 0.13 for other anaplastic gliomas). Median times to tumor progression were 31 and 32 weeks for patients with glioblastoma multiforme; 25th percentile times to progression were 70 and 40 weeks for patients treated with PCV and BCNU, respectively. For patients with other anaplastic gliomas treated with PCV and BCNU, median times to progression were 123 and 77 weeks, respectively. Multivariate analysis showed that the prognostic variables of age and Karnofsky scores were important for patients with glioblastoma multiforme and other anaplastic gliomas, and that the extent of surgical resection was important for those with other anaplastic gliomas. PMID- 2991487 TI - Immunohistochemical investigation of lysozyme, lactoferrin, alpha 1-antitrypsin, alpha 1-antichymotrypsin and ferritin in parotid gland tumors. AB - Presence of lysozyme, lactoferrin, alpha 1-antitrypsin, alpha 1-antichymotrypsin and ferritin was examined by the immunoperoxidase method in 15 consecutive parotid gland tumors as well as in normal parotid gland tissue. Lysozyme and lactoferrin were detected in intercalated duct cells of normal tissue and in the epithelial component of pleomorphic adenomas. alpha 1-antitrypsin, alpha 1 antichymotrypsin and ferritin were found in both epithelial and mesenchymal components of pleomorphic adenomas but not in normal parotid tissue. In the epithelial component of adenolymphoma only alpha 1-antichymotrypsin and lactoferrin were observed. The results would support a tentative histogenetic link between the intercalated duct cell and the epithelial component of the pleomorphic adenoma. PMID- 2991488 TI - Tumours of the minor (oropharyngeal) salivary glands: a demographic study of 336 cases. AB - There are 2410 primary epithelial salivary gland tumours in the files of the British Salivary Gland Tumour Panel. Of these tumours, 336 (14%) involved the minor (oropharyngeal) salivary glands, and these were studied in the present investigation. Individual tumours were diagnosed according to the WHO Classification. The percentage of malignant or potentially malignant tumours (46%) was much higher than in major glands (18%), and in some of the less common intraoral sites all the tumours were malignant. The principal sites were the palate (54%), lips (21%) and buccal mucosa (11%), and, in these sites, pleomorphic adenoma was the most common tumour. Monomorphic adenomas accounted for 6% of palatal tumours, but 30% of lip salivary gland tumours. The most common malignant tumour was the adenoid cystic carcinoma. The results are compared with several other large surveys and with tumours of major salivary glands. PMID- 2991489 TI - Rotavirus antibodies in the mother and her breast-fed infant. AB - The transfer of rotavirus antibodies from 25 healthy mothers to their breast-fed infants was investigated during the period of lactation (mean, 3.9 months; range, 1-9 months). Furthermore, the destiny of these antibodies in the infants' gastrointestinal tract and serum was examined. Rotavirus-specific immunoglobulins were analyzed by the ELISA (enzyme-linked immunosorbent assay) technique. All the mothers had rotavirus IgA and IgG in serum. About 80% of the mothers had low concentrations of rotavirus ScIg (i.e., antirotavirus immunoglobulin containing secretory component) in serum at the beginning of the lactation period declining to about 45% at the end of the period. From a few days after delivery to about 2 weeks later, the concentrations of rotavirus IgA and ScIg in milk declined. Thereafter, they remained unchanged. There was a positive correlation among the concentrations of rotavirus IgA in serum and rotavirus IgA as well as ScIg in milk. Rotavirus IgG in the infants' serum correlated with that of the mothers. Few samples of the infants' duodenal fluid contained rotavirus IgA or ScIg. On the other hand, about 80% of the infants' fecal samples contained rotavirus ScIg and IgA. Rotavirus IgA and ScIg disappeared from the infants' feces after cessation of lactation. Hence, it may be concluded that infants receive rotavirus IgG through the placenta, and rotavirus ScIg and IgA in constant amounts via milk throughout the period of lactation. The small intestine is flushed with rotavirus ScIg and IgA at each breast-meal, and these antibodies survive proteolysis in the gut. A possible protectional effect of rotavirus ScIg or IgA requires frequent breast-meals, and the effect is limited to the period of lactation. PMID- 2991490 TI - Root resorption associated with placement of a ceramic implant. Report of a case. AB - Ceramic grafting material was used to treat a vertical osseous defect associated with a maxillary left lateral incisor tooth. The area was monitored at maintenance appointments. Root resorption was detected radiographically 12 months after grafting. At 18 months, resorption was evident clinically in the presence of moderate inflammation. PMID- 2991491 TI - Longitudinal evaluation of durapatite ceramic as as alloplastic implant in periodontal osseous defects after 3 years. PMID- 2991492 TI - Analysis of solid-state Carbon-13 NMR spectra of polymorphs (benoxaprofen and nabilone) and pseudopolymorphs (cefazolin). AB - The solid-state 13C-NMR spectra of the polymorphs of benoxaprofen, nabilone, and cefazolin are reported using the cross-polarization/magic-angle spinning (CP/MAS) technique. In general, the spectra of the different crystal forms are different. In favorable cases the spectra of the drug in a pharmaceutical granulation can be discerned. These results provide a preliminary indication that solid-state NMR spectroscopy is a useful technique for the investigation of drug polymorphs and drugs in their dosage forms. PMID- 2991494 TI - Periungual angiofibroma. PMID- 2991493 TI - Beta-carbolines as benzodiazepine receptor ligands II: Synthesis and benzodiazepine receptor affinity of beta-carboline-3-carboxylic acid amides. AB - Numerous beta-carboline-3-carboxamides were synthesized by amidation of beta carboline-3-carboxylic acid, with various amino acids and amino acid esters serving as amine components, and tested in respect to their affinity for the benzodiazepine receptor in mouse brain membranes. The title compounds have affinities in the low micromolar range. The results are discussed with respect to their relevance for a possible beta-carboline structure containing the endogenous ligand of the benzodiazepine receptor. PMID- 2991495 TI - Molecular heterogeneity of leukotriene receptors: correlation of smooth muscle contraction and radioligand binding in guinea-pig lung. AB - The [3H]leukotriene C4 ([3H]LTC4) and [3H]leukotriene D4 ([3H] LTD4) specific binding sites in guinea-pig lung membranes were characterized and correlated with smooth muscle contractile activities of a series of LTC-, D- and E-type analogs. [3H]LTC4 bound to the specific sites with high affinity (dissociation constant Kd = 15 +/- 5 nM), saturable capacity (maximum binding = 68 +/- 15 pmol/mg of membrane protein), stereoselectivity and specificity. The [3H]LTC4 specific binding sites were detected in the membranes isolated from leukotriene sensitive (e.g., lung and heart) or insensitive (e.g., brain and red blood cells) tissues. [3H] LTD4 also bound to specific sites with high affinity (Kd = 0.20 +/- 0.05 nM), low capacity (maximum binding = 1.1 +/- 0.2 pmol/mg of membrane protein) stereoselectivity and specificity. The [3H] LTD4 specific binding sites were detected in the membranes isolated from lung and trachea. [3H]LTC4 specific binding was inhibited by treatment of the membranes with the sulfhydryl alkylating agent N-ethylmaleimide. [3H]LTD4 specific binding was more sensitive to heat treatment and p-hydroxymercuribenzoate than the [3H]LTC4 specific binding. Radioligand competition activities of the LTD- and LTE-type analogs correlated well with the agonist and antagonist smooth muscle contractile activities. In contrast, the radioligand competition activity of the LTC-type analogs did not correlate with smooth muscle contractile activities. These results indicate that the [3H]LTC4 and [3H]LTD4 specific binding sites in guinea pig lung membranes are chemically and physically distinct. The [3H]LTD4 specific binding sites represent physiologically and pharmacologically important receptors, and the smooth muscle contraction induced by LTD-, and possible LTE-, type analogs are mediated through the LTD4 receptors. PMID- 2991496 TI - Reserpine-induced postjunctional supersensitivity in rat vas deferens and caudal artery without changes in alpha adrenergic receptors. AB - Caudal artery and vas deferens from rats treated chronically with reserpine (1 mg/kg/day i.p.) were used to study drug-induced postjunctional supersensitivity in smooth muscle. There was no change in contractile sensitivity of the rat vas deferens after 1 day of reserpine treatment; however, there were similar increases in sensitivity 4 and 7 days after reserpine treatment. The potencies of phenylephrine, methacholine and potassium chloride in causing contraction of the vas deferens were significantly increased by 4.9-, 19.5- and 1.23-fold, respectively, after 7 days of chronic treatment. This treatment also increased the potencies of phenylephrine, serotonin and potassium chloride in causing contraction of rat caudal artery by 1.8-, 1.7- and 1.23-fold, respectively; however, the potency of clonidine was unchanged after 7 days of reserpine treatment. There was no change in sensitivity to phenylephrine 1 day after reserpine treatment but sensitivity was significantly increased after 4 days. There was no significant change in maximum contractile response in either tissue to any of these agents after chronic reserpine treatment. Scatchard analysis of saturation isotherms of specific [125I]BE 2254 binding to membrane fractions from rat vas deferens and caudal artery showed no change in the density or affinity of alpha-1 adrenergic receptors after 1, 4 or 7 days of chronic reserpine treatment. In addition, no change was observed in specific [3H]rauwolscine binding to either tissue after 7 days of chronic reserpine treatment, suggesting no change in alpha 2 adrenergic receptors. These data indicate that changes in alpha adrenergic receptors are not involved in postjunctional supersensitivity of smooth muscle caused by chronic reserpine treatment. PMID- 2991497 TI - Kinetics of captopril- and enalapril-induced inhibition of pulmonary angiotensin converting enzyme in vivo. AB - The kinetics of angiotensin converting enzyme (ACE) inhibition by captopril (SQ 14225) and enalapril (MK 421) in anesthetized rabbits was investigated. Kinetic parameters, apparent Km, an index of enzyme-substrate affinity and apparent Vmax, a measure of maximal rate of substrate conversion, were determined from indicator dilution measurements of single pass pulmonary metabolism of a synthetic ACE substrate [3H]benzoyl-phenylalanyl-alanyl-proline. Two methods for determination of kinetics in vivo from metabolism data were used. One fit pulmonary venous outflow metabolism data to a nonlinear model of saturable lung metabolic processes. This method required injection of sufficient substrate (benzoyl phenylalanyl-alanyl-proline) to produce a large range of intravascular substrate concentrations. An alternative method required use of only low intravascular substrate concentrations. Both methods rely primarily on similar Michaelis-Menten assumptions and generated very similar results. Both captopril (10 and 20 nmol/kg) and enalapril (4 and 7 nmol/kg) behaved as noncompetitive ACE inhibitors in vivo. ACE inhibition was characterized by depressed [3H]benzoyl-phenylalanyl alanyl-proline hydrolysis and apparent Vmax whereas apparent Km was unaffected. Other studies have suggested that these inhibitors act as competitive or mixed competitive and noncompetitive ACE inhibitors in vitro. Significant differences, however, between in vivo and in vitro experimental conditions suggest that the kinetics of enzyme inhibition in vitro may not necessarily reflect the action of the inhibitor in vivo. Additionally, results obtained in vivo may more accurately reflect the therapeutic behavior of these compounds. PMID- 2991498 TI - Stimulation of vasopressin release by gamma-aminobutyric acid antagonists in spinal cord transected rats. AB - Picrotoxin, a gamma-aminobutyric acid (GABA) antagonist, administered to spinal rats elicited dose-related increases in mean blood pressure and circulating plasma vasopressin concentration which were found to be highly correlated (r = 0.952; P less than .001) 6 min after infusion of picrotoxin. Pretreatment with the vasopressin antagonist d(CH2)5Tyr(Me)arginine vasopressin (10 microgram/kg i.v.) blocked the picrotoxin-induced pressor response. Administration of bicuculline (1.0 mg/kg i.v.), a second GABA antagonist, caused an increase in mean blood pressure and plasma vasopressin, whereas strychnine, another central nervous system stimulant thought not to act via a GABAergic mechanism, failed to evoke a significant change in either mean blood pressure or plasma vasopressin. Midcollicular decerebration decreased base-line plasma vasopressin concentrations and also prevented the picrotoxin-induced increase in pressure and vasopressin. The data from this study suggest that blockade of tonic GABAergic inhibition by GABA antagonists causes the release of vasopressin into the systemic circulation which results in a pressor response in spinal rats. The level at which this GABAergic inhibition occurs is not known; however, the GABA antagonists appear to require an intact supraspinal neuraxis to cause the release of vasopressin from the neurohypophysis. PMID- 2991499 TI - Characterization of a retinal melatonin receptor. AB - Melatonin (5-methoxy-N-acetyltryptamine) at picomolar concentrations (IC50, 40 pM) inhibited the calcium-dependent release of [3H]dopamine elicited at 3 Hz (2 min, 20 mA, 2 msec) from rabbit retina through activation of a site possessing the pharmacological and functional characteristics of a receptor. The effect of melatonin shows biological specificity as this hormone does not modify [3H]dopamine release from striatum or olfactory tubercle. This paper describes the effects of small modifications of the melatonin structure on the inhibition of calcium-dependent release of [3H]dopamine from retina. The more active melatonin analogs were those possessing a 5-methoxy group on carbon 5 of the indole nucleus and an N-acetyl group on the same position as in melatonin. The potencies of 5-methoxy indoles compounds was as follows (IC50): melatonin (40 pM) = 6-chloromelatonin (40 pM) greater than 6-hydroxymelatonin (1.6 nM) greater than or equal to 6-methoxymelatonin (2 nM) greater than 5-methoxytryptamine (63 nM) greater than 5-methoxy-N,N-di-methyltryptamine (200 nM) much greater than 5 methoxytryptophol (4 microM). The structure activity relationships of melatonin and related indoles indicated that the efficacy of melatonin is determined by the moiety substituted on carbon 5 (i.e., 5-methoxy group), whereas the affinity for the receptor is determined primarily by the moiety substituted on carbon 3 (i.e., ethyl N-acetyl group) of the indole nucleus. N-acetyltryptamine competitively antagonized the inhibitory effect of melatonin in the chicken retina and appears to be a partial agonist in the rabbit retina.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991500 TI - Rapid down regulation of beta adrenergic receptors by combining antidepressant drugs with forced swim: a model of antidepressant-induced neural adaptation. AB - The hypothesis that behavioral responses to antidepressant drugs in the forced swim test are related to a rapid neural adaptation produced by the combination of drug treatment and swim stress was explored. As a measure of adaptation, brain beta adrenergic receptors were assayed using [3H]dihydroalprenolol [( 3H]DHA) binding to brain membranes from rats that were processed in the forced swim test. The combination of swim stress and imipramine treatment antagonized immobility induced by forced swimming and resulted in a reduction in [3H] DHA binding to membranes from forebrain preparations which did not include the corpus striatum. Administration of antidepressant drugs from other chemical classes, including pargyline, iprindole and nomifensine, also reduced immobility induced by the forced swim and produced a reduction in [3H]DHA binding to forebrain membranes. In homogenates of the corpus striatum, [3H]DHA binding was not altered by swim stress combined with antidepressant drug treatment. Chlordiazepoxide was without an effect on immobility or beta receptor binding when combined with forced swim. Even though atropine and amphetamine exhibited a positive activity in the forced swim test, they did not reduce [3H]DHA binding. Therefore, by combining behavioral and neurochemical analysis of animals processed in the forced swim test, it may be possible to differentiate, with greater confidence, potential antidepressant drugs from "false positives." The present studies support the hypothesis that antidepressant drug action in the forced swim test involves a rapid neural adaptation as reflected by the down regulation of beta adrenergic receptors. Thus, this behavioral paradigm may serve as a model of adaptive mechanisms induced by antidepressant drugs. PMID- 2991501 TI - Effect of imipramine and adrenocorticotropin administration on the rat brain norepinephrine-coupled cyclic nucleotide generating system: alterations in alpha and beta adrenergic components. AB - Continuous treatment (1-3 weeks) with imipramine or adrenocorticotropin (ACTH) decreases the responsiveness of the norepinephrine-coupled cyclic nucleotide generating system in rat brain cerebral cortex. Experiments were undertaken to determine which component of the second messenger system is influenced by the hormone and antidepressant. Neither treatment modified the amount or function of extractable stimulatory guanine nucleotide binding protein or the activities of adenylate cyclase or phosphodiesterase. While both imipramine and ACTH treatment decreased the cyclic AMP response to norepinephrine, only imipramine administration influenced the response to isoproterenol. ACTH treatment was found to reduce the alpha adrenergic potentiation of isoproterenol- and 2 chloroadenosine-stimulated cyclic AMP production, as well as reduce the sensitivity of the norepinephrine response to prazosin. These findings indicate that imipramine and ACTH treatments decrease the responsiveness of the rat brain norepinephrine-stimulated cyclic AMP generating system through actions on the alpha and beta adrenergic receptor components. The results suggest that noradrenergic receptor activity may be under the control of adrenal and/or pituitary hormones. PMID- 2991503 TI - Vasopressin-mediated protein phosphorylation in intact toad urinary bladder. AB - Endogenous protein phosphorylation was examined in intact 32P-labeled toad bladders in response to arginine vasopressin (AVP) and other agents under conditions where the bladders were undergoing the normal hydroosmotic response. AVP increased 32P incorporation into proteins with apparent MW of 17,000, 28,000 and 34,000 and decreased 32P incorporation into a protein with MW 15,500. The cyclic AMP analog 8-(p-chloro-phenylthio)-cyclic AMP mimicked the effects of AVP on 32P incorporation. AVP-dependent changes in protein phosphorylation were found to be specific for the epithelium of the bladder and were blocked by the antagonist d(CH2)5-D-TyrVAVP. AVP caused increased phosphorylation even in the absence of an osmotic gradient, but the AVP-mediated decrease in 32P content of the 15,500 MW band was observed only in the presence of an osmotic gradient. Isolated epithelial cells also displayed AVP-stimulated increases in 32P incorporation into the MW 17,000 and 34,000 phosphoproteins, but no decrease in 32P incorporation into the 15,500-dalton band. Phosphorylation of the MW 34,000 band was maximal within 3 min. These data are consistent with the hypothesis that physiological effects of AVP may, in part, be mediated by cyclic AMP-dependent phosphorylation of specific proteins. PMID- 2991502 TI - Diuresis and suppression of vasopressin by kappa opioids: comparison with mu and delta opioids and clonidine. AB - The effects of agonists at kappa, mu and delta opioid receptors were determined on vasopressin levels in water-deprived rats. Bremazocine and U-50, 488 (kappa agonists) markedly suppressed vasopressin levels, whereas morphine and methadone (mu agonists) and metkephamid (a mixed mu and delta agonist) did not suppress vasopressin levels. Likewise, clonidine, a potent diuretic in normally hydrated rats, did not clearly suppress vasopressin levels. Metkephamid was shown to have a modest diuretic effect in normally hydrated rats but not in water-deprived rats. The diuretic effects of bremazocine were blocked completely by simultaneous treatment with desmopressin, a synthetic vasopressin-like analog. Desmopressin also blocked the diuretic effect of a waterload, but only partially attenuated the diuretic effect of clonidine. These results support the hypothesis that kappa opioid agonists produce a diuretic effect by suppressing plasma levels of vasopressin, and at higher doses produce a pattern of urination similar to animals lacking vasopressin. By comparison, mu and delta opioid agonists have little effect on vasopressin levels in water-deprived rats. PMID- 2991504 TI - The role of cyclic nucleotides in modulation of the membrane potential of the Schwann cell of squid giant nerve fibre. AB - The role of cyclic nucleotides in mediating the effects of nicotine cholinergic receptors has been investigated in Schwann cells of the giant nerve fibre of the squid. Elevation of cyclic AMP levels in this preparation by means of the phosphodiesterase inhibitor, theophylline, by the diterpene adenylate cyclase activator, forskolin, and by cyclic nucleotide analogues mimics the action of activating the nicotinic cholinergic receptors in producing a long-lasting hyperpolarization of the membrane potential of the Schwann cell. Theophylline and forskolin also potentiate the effects of carbachol and of neural stimulation on the Schwann cell. The results suggest that the nicotinic receptor of the squid Schwann cell is likely to mediate its effects via a mechanism that activates adenylate cyclase. The results are discussed in terms of the role of cyclic AMP in the complex multistep interaction between the giant axon of the squid and its surrounding Schwann-cell layer. PMID- 2991505 TI - Long-term potentiation of synaptic acetylcholine release in the superior cervical ganglion of the rat. AB - The release of endogenous acetylcholine (ACh) from the in vitro rat superior cervical ganglion was measured by assaying the bathing medium. Simultaneously, synaptic transmission in the ganglion was assessed by recording post-ganglionic compound action potentials. A brief period of tetanic preganglionic stimulation (20 Hz for 20 s) induced a long-term potentiation of the post-ganglionic compound action potential. The same tetanic stimulation also consistently induced a long term potentiation of stimulated ACh release. Spontaneous (non-stimulated) ACh release was not enhanced after tetanic stimulation. The content of ACh in the ganglion was not measurably increased after tetanic stimulation. These results suggest that the long-term increase in synaptic efficacy is due, at least in part, to an increase in the amount of ACh released by the afferent impulse. PMID- 2991506 TI - Heterosynaptic changes accompany long-term but not short-term potentiation of the perforant path in the anaesthetized rat. AB - Brief high-frequency trains of electrical stimulation delivered to the perforant path result in long-term potentiation (l.t.p.) of field potentials recorded extracellularly from granule cells of the dentate gyrus. L.t.p. of the population spike is often disproportionately greater than l.t.p. of the population excitatory post-synaptic potential (e.p.s.p.). We have investigated the basis of this effect in rats anaesthetized with sodium pentobarbitone. A series of graded stimuli were given before and after tetanization of the perforant path. From data obtained in this way, we plotted stimulus-response curves, and the relation (E-S curve) between the slope of the population e.p.s.p. (E) and the amplitude of the population spike (S). Curves relating spike onset latency to the slope of the e.p.s.p. were also constructed. Tetanization of the combined medial and lateral components of the perforant path led to long-term changes in the relation between the e.p.s.p. and the population spike. For a given e.p.s.p., the corresponding population spike was of greater amplitude and earlier onset. This E-S potentiation was marked by a shift to the left of the E-S amplitude curve and a downward displacement of the E-S latency curve. Tetanization of the lateral component of the perforant path had two long-term effects on responses evoked by test stimuli to the untetanized medial component: (1) long-term depression of the medial e.p.s.p. and (2) long-term E-S potentiation. The net result of these two heterosynaptically induced effects was to leave unaltered information transfer across medial perforant path-granule cell synapses; for a given test volley the e.p.s.p. was smaller, but because of E-S potentiation the population spike remained relatively unaffected. Short-term potentiation, which has a time course of only a few minutes and is presumed to be mediated by presynaptic mechanisms, was not accompanied by E-S potentiation or by corresponding changes in spike latency. Possible mechanisms of long-term heterosynaptic depression of the e.p.s.p. and of homo- and heterosynaptic E-S potentiation, are discussed. We conclude that although these effects probably reflect a generalized post-synaptic change, this change is unlikely to be a prolonged reduction in the membrane potential of granule cells. PMID- 2991507 TI - The cause of diverticular disease of the colon, its symptoms and its complications. Review and hypothesis. PMID- 2991508 TI - Embolisation of a massively bleeding hepatoma. A delayed sequel to needle biopsy. PMID- 2991509 TI - Granular cell myoblastoma of the breast. PMID- 2991510 TI - Changes in calmodulin level and cAMP-dependent protein kinase activity during epididymal maturation of ram spermatozoa. AB - Calmodulin concentration and cAMP-dependent protein kinase activity were simultaneously determined on ram spermatozoa collected by cannulation of successive segments of the epididymal tubule. Epididymal transit was characterized on one hand by an overall decrease in the calmodulin level and on the other by a dramatic rise in the cAMP-dependent protein kinase activity. In contrast to the calmodulin level, the cAMP-dependent protein kinase activity was correlated with the acquisition of flagellar beat. No further alterations in the level of these two proteins could be detected as spermatozoa acquired progressive motility. PMID- 2991511 TI - Pituitary and gonadal function in hypogonadotrophic hypogonadal (hpg) mice bearing hypothalamic implants. AB - GnRH receptor values are 30-50% of normal in pituitaries of hpg male mice, and testicular LH receptors only 8% of normal (160.4 +/- 17.6 and 2013 +/- 208.1 fmol/testis respectively). In male hpg mice bearing fetal preoptic area (POA) hypothalamic implants for 10 days there was no change in pituitary GnRH receptors, pituitary gonadotrophin content, or seminal vesicle weight. However, testicular weights and LH receptors were doubled in 4/10 mice and 2 had increased serum FSH levels. Between 26 and 40 days after implantation pituitary GnRH receptors and pituitary LH increased to normal male levels, although at 40 days serum and pituitary FSH concentrations had reached only 50% of normal values. Testicular and seminal vesicle weights increased more than 10-fold by 40 days after implantation and LH receptors to 70% of normal. In hpg female mice bearing hypothalamic implants for 30-256 days pituitary gonadotrophin concentrations were normal, even though GnRH receptors reached only 60% of normal values (6.18 +/- 0.4 and 9.8 +/- 0.4 fmol/pituitary respectively). Serum FSH was substantially increased from values of less than 30 ng/ml in hpg mice to within the normal female range in hypothalamic implant recipients. Ovarian and uterine weights increased after hypothalamic grafting from only 4-5% to over 74% of normal values. LH receptors increased from 6.5 +/- 1.3 fmol/ovary for hpg mice to 566.9 +/- 39.2 fmol/ovary for implant recipients. Vaginal opening occurred about 23 days after implantation and these animals displayed prolonged periods of oestrus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991512 TI - Effect of forskolin on maintenance of meiotic arrest and stimulation of cumulus expansion, progesterone and cyclic AMP production by pig oocyte-cumulus complexes. AB - Forskolin induced biphasic responses of cumulus progesterone secretion (determined by RIA) and cumulus mass expansion, with maximal increases occurring at 6.25 microns, and subsequent dose-dependent declines observed up to 10 microns forskolin. The diterpene induced dose-dependent responses in the % germinal vesicle (GV) of cumulus-enclosed and denuded oocytes (0.23 and 4.84 microns maintained 50% GV, respectively), it increased the cAMP content of cumulus masses, cumulus-enclosed oocytes and denuded oocytes, and increased heterologous metabolic coupling (determined by measuring transfer of radiolabelled uridine marker from the cumulus mass to the oocyte). A significant correlation was established between the amount of cAMP within the cumulus mass and that in the corresponding oocyte (r = 0.58). Above 10 microns-forskolin, the cAMP content of cumulus-enclosed oocytes was significantly greater than that of denuded oocytes (100 microns-forskolin: 0.118 +/- 0.082 and 0.006 +/- 0.001 pmol/oocyte respectively; P less than 0.001, paired t test), and the enhanced arresting action of forskolin upon cumulus-enclosed oocytes was correlated with an increase in intra-oocyte cAMP. Maintenance of meiotic arrest and stimulation of oocyte cumulus cAMP were reversible. During 48 h of culture, the arresting action of forskolin (50 microns) was maintained on denuded and cumulus-enclosed oocytes but heterologous metabolic coupling significantly declined. The cAMP content of the cumulus mass and corresponding oocyte significantly declined, while that of the denuded oocyte remained unchanged. The cAMP content of arrested cumulus-enclosed oocytes cultured for 48 h in 50 microns-forskolin was significantly greater than that of maturing oocytes cultured for 24 h in 50 microns-forskolin and then for 24 h in control medium. These results show that (1) forskolin stimulates progesterone secretion and expansion of pig cumuli, but at high doses the drug inhibits these functions while cumulus cAMP remains elevated; (2) when heterologous metabolic coupling is maintained, cumulus cAMP may be transferred to the oocyte; (3) the pig oocyte can synthesize cAMP; and (4) forskolin-maintenance of meiotic arrest of pig oocytes is correlated with elevated intra-oocyte cAMP but a 'factor' other than cAMP is also involved in maintenance of meiotic arrest. PMID- 2991513 TI - Diagnosis of genital herpes simplex virus infections. AB - Genital herpes simplex virus (HSV) infections have increased markedly in the last ten years. The clinical distinction between HSV and other infectious and noninfectious causes of genital ulcer can be difficult. As such, laboratory confirmation of HSV infection should be used for all women who present with ulcerative or clinically atypical genital lesions. Isolation of the virus in tissue culture remains the gold standard for the laboratory diagnosis of HSV infections. However, many new diagnostic methods of detecting HSV antigens in clinical specimens have been developed. Those procedures appear most useful in specimens taken from genital lesions, and at present they are unproven as reliable methods of obstetric management. Serologic methods can be useful in detecting past HSV infection; however, serodiagnosis is a less sensitive and timely procedure in patients with acute HSV infection. PMID- 2991514 TI - AIDS virus infection: prognosis and transmission. PMID- 2991515 TI - Treatment of cigarette smoking with short-term high-dosage corticotrophin therapy: preliminary communication. AB - Uncontrolled studies have indicated that the smoker's "life' can be simulated clinically by short-term, high-dosage corticotrophin therapy. Following such treatment, smokers previously prevented by the withdrawal syndrome from giving up smoking were able to do so easily. All 10 patients reported here were still not smoking after seven months. PMID- 2991516 TI - Diplopia: an unusual presentation of insulinoma. PMID- 2991518 TI - Synthesis and biological activity of carboxylic acid replacement analogues of the potent angiotensin converting enzyme inhibitor 5(S)-benzamido-4-oxo-6 phenylhexanoyl-L-proline. AB - The carboxylic acid group on the proline of 1 was replaced by a phosphoric acid, a hydroxamic acid, and a tetrazole to give compounds 2-4, respectively. Testing of 2-4 as angiotensin converting enzyme (ACE) inhibitors gave I50 values of 100, 1.6, and 22 microM, respectively, compared to 0.07 microM for 1. A hydroxamic acid derivative of the ketomethylene pentapeptide analogue 18 was then synthesized. This compound, 17, had an ACE I50 of 0.011 microM compared to 0.0076 microM for 18. Oral administration of 10 mg/kg of 17 to renal hypertensive rats had no effect on blood pressure or heart rate. PMID- 2991517 TI - Synthesis and biological activity of pentapeptide analogues of the potent angiotensin converting enzyme inhibitor 5(S)-benzamido-4-oxo-6-phenylhexanoyl-L proline. AB - Two pentapeptide analogues of the ketomethylene-containing angiotensin converting enzyme (ACE) inhibitor 5(S)-benzamido-4-oxo-6-phenylhexanoyl-L-proline (1) were synthesized and evaluated as ACE inhibitors and antihypertensive agents. Compounds 14 and 15 were very potent ACE inhibitors with I50 values of 7.0 and 3.0 nM, respectively, compared to an I50 value of 70 nM for 1. Neither 14 nor 15 showed significant blood pressure lowering activity in renal hypertensive rats. Investigations conducted on a tritiated analogue of 14 showed that 70% of an oral dose of this compound is absorbed but is rapidly excreted from the blood with a half life of 24 min. Thin-layer chromatography of bile and urine contents in rats given tritiated 14 orally showed that it is excreted in greater than 90% unchanged form. This implies that a ketomethylene linkage can stabilize peptide amide linkages adjacent to it to peptidase degradation. PMID- 2991519 TI - Synthesis of xanthines as adenosine antagonists, a practical quantitative structure-activity relationship application. AB - A set of 56 8-phenylxanthines, previously tested for adenosine antagonism (adenosine A1 receptor affinity), was analyzed by quantitative structure-activity relationship (QSAR) techniques. The resulting QSAR revealed that (1) the most potent receptor binders had already been made in this series and thus suggested the termination of synthesis of compounds with additional phenyl substituents to increase potency and (2) potency was much more strongly affected by changes in ortho than para phenyl substitution. On the basis of this study, an additional 20 compounds were synthesized that contained primarily para substituents designed to increase aqueous solubility. High potency was maintained among the resulting sulfonamide derivatives (as predicted by the QSAR), and aqueous solubility was dramatically increased. Furthermore, in vitro antagonism of an adenosine receptor mediated physiological effect was demonstrated. PMID- 2991520 TI - Optimization of the Schiff bases of N-hydroxy-N'-aminoguanidine as anticancer and antiviral agents. AB - Hydroxyurea, hydroxyguanidine, and some thiosemicarbazones have been shown to have anticancer and antiviral activities. One of their possible sites of action is the enzyme ribonucleotide reductase (RR). Combination of the structural features of these compounds led to the design and synthesis of the Schiff bases of N-hydroxy-N'-aminoguanidine. Synthesis and structure-activity studies of some of these compounds point to increased size and lipophilicity as important factors for activity. To optimize the activities of this series of compounds, 13 derivatives of high lipophilicity and molecular size have been synthesized and their biological activities studied. The most active anticancer compounds against L1210 in vitro (compounds 9 and 12) are about 7 times more active than hydroxyguanidine and hydroxyurea. The most active antiviral compounds against Rous sarcoma virus transformation of chick fibroblasts in vitro (7, 9) are about 40 times more active than hydroxyguanidine. One of the compounds (4) shows promising activity in vivo (% T/C = 140 against P388 leukemia in mice) and is undergoing further studies by the National Cancer Institute (NCI). Studies of the inhibition of transformation of chick embryo cells by Rous sarcoma virus show that all these compounds inhibit transformation while some compounds inhibit viral replication as well. PMID- 2991521 TI - Synthesis and pharmacological evaluation of cis-2,3,3a,4,5,6,7,7a-octahydro-3 oxoisoxazolo[5,4-c]pyridine: a structural analogue of the GABA agonist THIP. AB - The pharmacological activities of the GABA agonist muscimol (1) and its dihydro analogue (2) have been shown to be almost identical. The closely related 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP, 3), although biologically less active than muscimol, was selected for clinical trials. We report the synthesis of the so far elusive cis-2,3,3a,4,5,6,7,7a-octahydro-3-oxoisoxazolo[5,4 c]pyridine (cis-DH-THIP, cis-4), which--surprisingly--is devoid of any GABAergic activity. PMID- 2991522 TI - Modifications on the heterocyclic base of acyclovir: syntheses and antiviral properties. AB - A group of compounds was prepared in which variations of the ring portion of the acyclovir (ACV) structure were made. These modifications included monocyclic (isocytosine, triazole, imidazole), bicyclic (8-azapurine, pyrrolo[2,3 d]pyrimidine, pyrazolo[3,4-d]pyrimidine) and tricyclic (linear benzoguanine) congeners. The derivatives were evaluated against herpes simplex virus type 1 (HSV-1) by the plaque-inhibition and plaque-reduction methods with only the 8 azapurine analogue 28 showing some activity. In a test measuring the ability of these compounds to inhibit the HSV-1 thymidine kinase, 28 and the tricyclic derivative 38 exhibited competition with ACV for binding to the enzyme. The inability of the group to exert significant antiherpetic action is attributed to their lack of phosphorylation to the requisite triphosphate stage. PMID- 2991523 TI - Immunity to varicella-zoster virus in a normal adult population. AB - Sera from 489 trainee nurses were examined, by the ELISA technique, for the presence of varicella-zoster virus specific antibody; antibody was found in 446 (91.2%). In more detailed investigations of specific immunity in 33 healthy adults with a past history of chickenpox, 32 (97%) showed a positive lymphocyte transformation test, but only 11 out of 23 examined (48%) demonstrated mononuclear cell production of specific antibody in vitro; serum antibody was found in 30 (91%) by the ELISA and in 25 out of 26 examined (96%) by the FAMA technique. A high degree of correlation was found between both a positive lymphocyte transformation and the presence of serum antibody and a past history of chickenpox. PMID- 2991524 TI - Morphological response of human rotavirus to ultra-violet radiation, heat and disinfectants. AB - The morphological damage induced in human rotavirus particles by exposure to ultraviolet (UV) radiation at a wavelength of 254 nm increased progressively with length of treatment. Exposure of the virus in suspension to 9000 ergs/cm2/s was sufficient to remove the smooth capsid layer from 50% of particles after 1 min and from all the virions within 10 min. By this time, the number of stain penetrated or empty particles increased markedly, along with the appearance of virus-derived debris in the form of disrupted and isolated capsomeres. After treatment for 120 min no intact virus particles were observed. The action of wet (100 degrees C) or dry (60 degrees C) heat resulted in changes similar to those effected by UV radiation, with a rapid loss of viral outer capsid shell from the virions followed by stain penetration and disintegration of particles. Sodium hypochlorite, cetrimide and 70% ethanol induced a rapid loss of the outer capsid layer, but, compared with UV radiation or heat, a slower increase in the number of stain-penetrated particles was noted. This was particularly evident with cetrimide. Chlorhexidine and phenol had effects on virus structure only after extended periods of exposure, whilst glutaraldehyde treatment had little influence on virus morphology. Glutaraldehyde 2% v/v would appear to be most suitable for the disinfection of rotavirus-containing electronmicroscope grids before their examination. PMID- 2991525 TI - Restriction endonuclease digestion analysis of DNA from viruses isolated from different sites of two fatal cases of herpes simplex virus type-1 infection. AB - Herpes simplex virus (HSV) type-1 was isolated from a fatal case of herpes simplex encephalitis (case 1) and from a fatal case of disseminated herpes simplex (case 2). The virus was isolated from the lip lesion, the frontal lobe and the temporal lobe of the brain in case 1 and from a mesenteric node, myocardium and salivary gland in case 2. Restriction endonuclease digestion analysis showed that each case was infected with different substrains of HSV. The changes in band pattern in isolates from case 1 occurred in the "variable" region of the genome, showing that viruses with such variations can be isolated simultaneously from different tissues. The changes in band patterns in isolates from case 2 indicated the presence of two virus substrains, one in the mesenteric node and salivary gland and a second in the myocardium. PMID- 2991526 TI - Characterisation of cryptic plasmids in clinical isolates of Bacteroides fragilis. AB - A screening method for plasmids in the fragilis group of Bacteroides spp. was developed, taking account of the lysozyme resistance of these species; 26 strains, 24 of them B. fragilis, were investigated by this method. Eleven strains contained plasmids and up to three different plasmids were found in individual strains. The plasmids belonged to five different size classes of mol. wt (10(6] 2.8, 3.5, 3.6, 4.2 and 19. Plasmids of equal size showed homology; no homology was found between plasmids of different sizes. Plasmids of equal size showed identical restriction patterns with 17 restriction endonucleases. Restriction maps were constructed for the five classes of plasmid. PMID- 2991527 TI - Immunological effect of silica dust analyzed by monoclonal antibodies. AB - We found a significant decrease of OKT8 + ve cells in silicosis patients (18.1%), but also in unaffected exposed workers (19.0%), when compared with sex- and age matched controls (22.8%). The proportion of OKT8 + ve cells was significantly lower in subjects with antinuclear antibodies (15.7%) and in those with IgG rheumatoid factors (16.3%). PMID- 2991530 TI - Organization of the Dictyostelium discoideum actin multigene family. Flanking sequences show subfamily homologies and unusual dyad symmetries. AB - Sequences flanking the protein-coding regions of 15 of the 17 to 20 actin genes in the cellular slime mold Dictyostelium discoidium have been determined. Comparison of sequences among genes shows that they contain extensive homologies at both the 5' and 3' ends of the coding regions. On the basis of these homologies, actin genes fall into three groups. Group I consists of Actin 8 alone. Group II consists of the two closely linked genes Actin 3-sub1 and Actin 3 sub2. These two genes differ from all other actin genes in the location of their TATA box and oligo(dT) run, and diverge substantially in their coding sequence as well. Group III contains all the rest of the genes we have studied. Within this group, there are two subgroups of genes, IIIA (Actins 5, 9 and 10) and IIIB (Actins M6, 2-sub1 and 2-sub2, 4, 6, 7, 11 and 12). Two actin cDNA clones, ITL-1 and III-12/A1, which have no cloned genomic counterparts, are members of groups IIIA and IIIB, respectively. Homologies at the 3' ends of genes do not extend beyond a short genomic poly(A) sequence, the probable termination of transcription. Homologies at the 5' ends may extend about 300 base-pairs 5' to the ATG but, in most cases, extend only about 150 base-pairs 5' to the ATG. We have identified a group of short, relatively G + C-rich sequences within the extremely A + T-rich sequence at the 5' ends of actin-coding regions, which are shared among different actin genes. Many of these sequences exhibit dyad symmetry, and their general location and order is conserved among the different actin genes. We suggest that they may have a role in regulation of the transcriptional patterns of individual actin genes. PMID- 2991528 TI - Optical study of active ion transport in lipid vesicles containing reconstituted Na,K-ATPase. AB - A fluorescence method is described for the measurement of ATP-driven ion fluxes in lipid vesicles containing purified Na,K-ATPase. The membrane voltage of enzyme containing vesicles was measured by using a voltage-sensitive indocyanine dye. By addition of valinomycin the vesicle membrane is made selectively permeable to K+ so that the membrane voltage approaches the Nernst potential for K+. With constant external K+ concentration, the time course of internal K+ concentration can be continuously measured as change of the fluorescence signal after activation of the pump. The optical method has a higher time resolution than tracer-flux experiments and allows an accurate determination of initial flux rates. From the temperature dependence of active K+ transport its activation energy was determined to be 115 kJ/mol. ATP-stimulated electrogenic pumping can be measured as fast fluorescence change when the membrane conductance is low (i.e., at low or zero valinomycin concentration). In accordance with expectation, the amplitude of the fast signal change increases with decreasing passive ion permeability of the vesicle membrane. The resolution of the charge movement is so high that a few pump turnovers can be easily detected. PMID- 2991529 TI - Functional reconstitution of lens gap junction proteins into proteoliposomes. AB - Membranes rich in junction complexes were prepared from bovine lens, and the fragments of the membranes were reconstituted into proteoliposomes with a large excess of phosphatidylcholine and dicetylphosphate. The osmotic swelling behavior of these liposomes showed that the lens junction membranes contributed protein components that produced channels with a nominal diameter of 1.4 nm. Most preparations of lens junctions produced rates of osmotic swelling much slower than those found in proteoliposomes containing equivalent amounts of Escherichia coli porin, and we discuss several possible explanations for this observation. PMID- 2991531 TI - Solution structure of mitochondrial cytochrome c. I. 1H nuclear magnetic resonance of ferricytochrome c. AB - The 1H nuclear magnetic resonance spectra of tuna and horse ferricytochromes c have been investigated and the resonances of all amino acid methyl groups have been assigned to specific absorption lines. The assignment procedure involves principally the comparison of one-dimensional nuclear magnetic resonance spectra from a range of homologous ferricytochromes c and does not require a prior knowledge of the secondary or tertiary protein structure. Of the 49 methyl groups of tuna cytochrome c, the assignment of 33 is made without reference to the X-ray crystal structure. The method should therefore be applicable to other proteins of similar size where X-ray structures are unavailable. The assignments will be used to investigate the structure of cytochrome c in solution. PMID- 2991532 TI - Solution structure of mitochondrial cytochrome c. II. 1H nuclear magnetic resonance of ferrocytochrome c. AB - The 1H nuclear magnetic resonance spectrum of tuna ferrocytochrome c has been studied and the resonances of all 49 amino acid methyl groups have been assigned to specific absorption lines. In comparison with resonance assignments in the ferricytochrome c spectrum, the secondary shifts of resonances of ferrocytochrome c are smaller and the identification of characteristic spin-systems from comparison of spectra from homologous proteins more difficult. For this reason, two-dimensional nuclear magnetic resonance exchange correlated spectroscopy has been used to correlate the assigned resonances of tuna ferricytochrome c with previously unassigned resonances of tuna ferrocytochrome c. PMID- 2991533 TI - Comparison of the solution and crystal structures of mitochondrial cytochrome c. Analysis of paramagnetic shifts in the nuclear magnetic resonance spectrum of ferricytochrome c. AB - The two accompanying papers describe the assignment of methyl-containing spin systems in the 1H nuclear magnetic resonance spectra of tuna ferricytochrome c and tuna ferrocytochrome c. At present, 104 resonances from 208 C-H protons are assigned in both oxidation states. In this paper, the difference in chemical shift of a resonance between the two oxidation states is used together with a dipolar model of the unpaired electron spin of ferricytochrome c to compare the structure of cytochrome c in solution with three high-resolution structures of cytochrome c obtained by X-ray diffraction in single crystals. The overall protein fold and the positions of most of the haem-packing residues are shown to be invariant between the crystal and solution. However, three regions of the protein, at the C terminus, around the haem propionic acid groups and at the haem crevice near thioether-2, are found to undergo conformational changes on the removal of crystal packing constraints. PMID- 2991534 TI - Molecular movements promoted by metal nucleotides in the heavy-chain regions of myosin heads from skeletal muscle. AB - Molecular movements generated in the heavy-chain regions (27-50-20(X 10(3)) Mr) of myosin S1 on interaction with nucleotides ATP, AMPPNP, ADP and PPi were investigated by limited proteolysis of several enzyme-metal nucleotide complexes in the absence and presence of reversibly bound and crosslinked F-actin. The rate and extent of the nucleotide-promoted conversion of the NH2-terminal 27 X 10(3) Mr and 50 X 10(3) Mr segments into products of 22 X 10(3) Mr and 45 X 10(3) Mr, respectively, were estimated to determine the amplitude of the molecular movements. The 22 X 10(3) Mr peptide was identified by amino acid sequence studies as being derived from cleavage of the peptide bond between Arg and Ile (at position 23 to 24). The 45 X 10(3) Mr peptide, previously shown to represent the NH2-terminal part of the 50 X 10(3) Mr region, would be connected to the adjacent C-terminal 20 X 10(3) Mr region by a pre-existing loop segment of about 5 X 10(3) Mr; the proteolytic sensitivity of the latter region is increased particularly by nucleotide binding. The tryptic reaction proved to be a sensitive indicator of the conformational state of the liganded heavy chain as the rate of peptide bond cleavage in the two regions is dependent on the nature of the bound ligand; it decreases in the order: ATP greater than AMPPNP greater than ADP greater than PPi. It depends also on the nature of the metal present, Mg2+ and Ca2+ being much more effective than K+. Binding of F-actin to the S1-MgAMPPNP complex affords significant protection against breakdown of 27 X 10(3) Mr and 50 X 10(3) Mr peptides, but with concomitant hydrolysis of the 50 X 10(3) Mr-20 X 10(3) Mr junction. Additionally, interaction of MgATP with HMM modulates the tryptic fission of the S1-S2 region. The overall data provide a molecular support for the two-state model of the myosin head and emphasize the involvement of the 50 X 10(3) Mr unit in the mechanism of coupling between the actin and nucleotide binding sites. PMID- 2991535 TI - Conservation of major nuclease S1-sensitive sites in the non-conserved spacer region of ribosomal DNA in Drosophila species. AB - We have analysed nuclease S1-sensitive sites in cloned ribosomal DNA repeats from Drosophila melanogaster, D. hydei and D. virilis. All species contain major S1 sensitive sites in the spacer near the region of transcription termination, albeit with somewhat different positions and sensitivities. The same sites are also sensitive to the single-strand specificity of Bal31 nuclease at neutral pH. Additional major sites exist at each end of the intervening sequence within the 28 S gene of non-transcribed intervening-sequence-positive ribosomal DNA units of D. hydei. Only minor sites, however, were detected in the Pol I promoter regions. This is in contrast to Pol II transcribed genes, where S1 hypersensitivity becomes apparent at the 5' ends during gene expression. We have sequenced and mapped the S1 sites in the D. hydei spacer. They consist mainly of alternating A and T nucleotides that could form small cruciform structures. Cross-hybridization at low stringencies between the relevant S1-sensitive spacer regions of the three species indicates that the sites lie within very divergent sequences. We discuss the potential functional significance of S1 sites in rDNA spacers and intervening sequences, and the manner in which they might be maintained during rDNA sequence divergence. PMID- 2991537 TI - The adrenochrome pathway: the major route for adrenalin catabolism by polymorphonuclear leucocytes. AB - Oxygen radical production by polymorphonuclear leucocytes stimulated the oxidation of adrenalin through the adrenochrome pathway. This was detected either spectrophotometrically at 480 nm or separated by hplc and detected radiochemically. The oxidation was detectable within 5 min and continued for at least 4 h. Over the adrenalin concentration range 0.3 microM to 10 mM more than 80% of the oxidation that occurred was through the adrenochrome pathway, the remainder being through the amine oxidase, catechol methyl transferase pathway. Medium isolated after stimulation of the polymorphonuclear leucocytes was also able to oxidise adrenalin to adrenochrome. The results provide a cellular mechanism for the formation of adrenochrome and the other metabolites on this pathway of adrenalin metabolism, in inflammatory conditions where polymorphonuclear leucocyte infiltration occurs. PMID- 2991536 TI - Binding of an antitumor drug to DNA, Netropsin and C-G-C-G-A-A-T-T-BrC-G-C-G. AB - The antitumor antibiotic netropsin has been co-crystallized with a double-helical B-DNA dodecanucleotide of sequence: C-G-C-G-A-A-T-T-BrC-G-C-G, and the structure of the complex has been solved by X-ray diffraction at a resolution of 2.2 A. The structure has been refined independently by Jack-Levitt and Hendrickson-Konnert least-squares methods, leading to a final residual error of 0.257 by the Jack Levitt approach (0.211 for two-sigma data) or 0.248 by the Hendrickson-Konnert approach, with no significant difference between refined structures. The netropsin molecule displaces the spine of hydration and fits snugly within the minor groove in the A-A-T-T center. It widens the groove slightly and bends the helix axis back by 8 degrees, but neither unwinds nor elongates the double helix. The drug molecule is held in place by amide NH hydrogen bonds that bridge adenine N-3 and thymine O-2 atoms, exactly as with the spine of hydration. The requirement of A X T base-pairs in the binding site arises because the N-2 amino group of guanine would demand impermissibly close contacts with netropsin. It is proposed that substitution of imidazole for pyrrole in netropsin should create a family of "lexitropsins" capable of reading G X C-containing base sequences. PMID- 2991538 TI - Influence of the diabetic state on isoproterenol-induced cardiac necrosis. AB - Isoproterenol-induced myocardial necrosis was examined in male mice with alloxan induced and genetically transmitted diabetes. Ten days after alloxan treatment, mice exhibited an elevation in blood glucose concentrations, weight loss, polyuria and decreased heart rates (510 +/- 15 v. 675 +/- 11 beats/min) compared with matched control mice. Similarly, genetically diabetic mice exhibited lower heart rates than the corresponding age-matched controls (383 +/- 30 v. 603 +/- 30 beats/min). In comparison to matched controls, both groups of diabetic mice had a significant decrease in the severity of the cardiac necrosis which was induced by the administration of isoproterenol. The reduction in isoproterenol-induced cardiac lesions was similar in mice with chemically induced diabetics and mice with genetically transmitted diabetes. Biochemical studies of ventricular slices revealed no change in basal cAMP levels and no differences in isoproterenol induced changes in cAMP levels in mouse hearts from both models of diabetes. Insulin treatment corrected the chemically induced diabetic state and restored the cardiotoxic potential of isoproterenol. PMID- 2991539 TI - Myocardial injury following endogenous catecholamine release in rabbits. AB - Catecholamines (CAT) given in large doses produce cardiomyopathic changes in several animal species. This study was designed to determine if endogenous release can also induce cardiac injury. Rabbits were infused with doses of tyramine (TYR), ranging from 200 to 500 micrograms/min/kg, i.v. for 90 min. Arterial pressure and heart rate were measured, as were total CAT concentrations, blood gases, pH and glucose. Two days later the animals were killed and cardiac injury assessed using a histological scoring system. All data were compared with controls given saline. Initial CAT averaged 452 pg/ml, rose to 2890 pg/ml after starting TYR, 500 micrograms/min/kg, and remained elevated for the duration of infusion. Circulating CAT levels were a function of TYR dose, and bore a linear relationship to the histological score (P less than 0.001). Development of lesions was unaltered by beta 1 blockade with practolol, but sharply reduced by alpha blockade with phentolamine (P less than 0.01). Pretreatment with insulin also reduced lesion formation, but diabetic (alloxan) rabbits showed no greater CAT injury. It is concluded that endogenous release of CAT induces myocardial injury in the rabbit in a dose-dependent manner. This is unrelated to myocardial O2 demand, and microvascular pathology was absent. Activation of alpha adrenergic pathways is likely the dominant or exclusive mechanism. PMID- 2991540 TI - The effects of trimetoquinol on the intact rabbit heart and myocardial adenylate cyclase activity: evidence for spare myocardial beta receptors. AB - We have compared and contrasted the actions of (-)isoproterenol and (+/-) trimetoquinol on rabbit heart preparations. In the presence of either GTP or Gpp[NH]p (guanosine-5'-(beta, gamma imino) triphosphate), trimetoquinol displayed partial agonist activity in stimulating adenylate cyclase activity in a particulate rabbit heart preparation. Trimetoquinol enhanced adenylate cyclase activity 20% or 65% of the maximum obtainable by isoproterenol in the presence of GTP or Gpp[NH]p respectively. In the presence of GTP, concentrations of catecholamines required to enhance cyclase activity 15% of the maximum obtainable with isoproterenol (EC15) were 2.0 X 10(-7) M and 5.5 X 10(-8) M for trimetoquinol and isoproterenol, respectively. In the presence of Gpp[NH]p EC30 values were 2.0 X 10(-7) and 3.5 X 10(-8) M for trimetoquinol and isoproterenol respectively. Trimetoquinol also displayed partial agonist activity for the ability to increase cAMP levels in the isolated perfused rabbit heart. By contrast trimetoquinol was equieffective to isoproterenol at increasing tension development and rate of contraction of the isolated perfused heart. Concentrations of catecholamines required to increase tension and rate of contraction 50% of the maximum obtainable with isoproterenol were 1.5 X 10(-7) M and 1.7 X 10(-8) M for trimetoquinol and isoproterenol, respectively. These data show that only a partial stimulation of adenylate cyclase activity and cAMP levels by trimetoquinol is sufficient to produce maximal changes in mechanical activity of the heart.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991541 TI - Immunological analysis of auto-antibodies against the adenine nucleotide translocator in dilated cardiomyopathy. AB - We have previously identified the adenine nucleotide translocator (ANT), an intrinsic protein of the inner mitochondrial membrane, as an auto-antigen in dilated cardiomyopathy (DCM). Further immunochemical characterization by crossed immunoelectrophoresis, indirect solid phase radioimmunoassay and immunoadsorption studies on the isolated translocator protein and mitochondria from heart, kidney and liver showed the existence of organ-specific antigenic determinants although partial crossreactivity between the three proteins was observed. Sera from 18 patients with histologically proven dilated cardiomyopathy were studied for their capacity to bind to the translocator protein. Seventeen of 18 patients showed significant binding, while in the sera of patients with coronary heart disease, suspected alcoholic heart disease or healthy blood donors, no anti-ANT antibodies were observed. Further studies showed organ-specific and functionally active autoantibodies, which decreased the ADP/ATP exchange rate from heart mitochondria. A close correlation was found between the antibody-titer and the hemodynamic function. These results give new evidence for autoimmunological events in dilated cardiomyopathy. PMID- 2991542 TI - Existence and functional role of alpha 1-adrenoceptors in the mammalian heart. PMID- 2991543 TI - The effect of verapamil on ischaemia-induced changes to the sarcolemma. AB - The effect of verapamil on sarcolemmal activities of sarcolemmal fragments isolated from aerobically perfused (control) and ischaemic rat hearts was examined. Adding verapamil to the perfusate of aerobically perfused hearts for 75 min enhanced some of the sarcolemmal activities; Na+-K+ ATPase (31%), K+ stimulated phosphatase (31%) and Na+-Ca2+ exchange rate (46%). Adding verapamil directly to the enzymatic incubation media, or to the cardiac homogenate prior to sarcolemmal isolation did not alter these activities, suggesting that these changes are dependent upon addition of verapamil to the intact system. Addition of verapamil to hearts 15 min prior to a 60 min ischaemic episode maintained a number of sarcolemmal activities close to those obtained after aerobic perfusion. Na+-K+ ATPase activity and Na+-Ca2+ exchange received a relative protection while K+ stimulated phosphatase activity was not protected. 5'-nucleotidase activity was completely protected against ischaemia-induced depression. The mechanism whereby verapamil induces these changes in sarcolemmal enzymatic activities is unclear but its ability to maintain these activities at or near normal levels may contribute to its ability to protect against the deleterious effects of ischaemia. PMID- 2991544 TI - The protective effect of prazosin on the ischaemic and reperfused myocardium. AB - Experiments were undertaken to establish whether prazosin prevents isolated hearts from gaining excess Ca2+ during post-ischaemic reperfusion, to determine whether this effect is dose-dependent, if it is accompanied by a change in the energy-rich phosphate reserves, and whether prazosin is effective when added only upon reperfusion. Isolated, spontaneously beating rat hearts were used. The ischaemic episodes ranged from 15 to 60 min, and prazosin (0.01 to 10 micro mol/1) was added both before inducing ischaemia and upon reperfusion. When 0.01 to 1 micro mol/1 prazosin was present before and after the ischaemic episode the reperfusion-induced gain in Ca2+ was attenuated, but not abolished. Pretreatment with 0.01 to 1 micro mol/1 prazosin slowed the ischaemic-induced rise in resting tension, enhanced mechanical recovery after 30 but not 60 min ischaemia, and exerted a dose-dependent slowing effect on the ischaemia-induced depletion of ATP and CP, with 1 micro mol/1 being the optimal dose. Adding 0.01 to 1 micro mol/1 prazosin at the time of reperfusion neither prevented excess Ca2+ accumulation upon reperfusion nor did it exert an energy-sparing effect. 5 to 10 micro mol/1 prazosin did not attenuate the reperfusion-induced gain in Ca2+, irrespective of whether it was added before or only at the time of reperfusion. These results show that the dose-response curve for the inhibitory effect of prazosin on Ca2+ overload is complex, and that adding prazosin coincident with the reperfusion of isolated ischaemic hearts does not attenuate Ca2+ gain. PMID- 2991545 TI - Developmental expression of 2',3'-cyclic nucleotide 3'-phosphohydrolase in dissociated fetal rat brain cultures and rat brain. AB - The development in primary dissociated rat brain cultures of 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) activity, the accumulation of CNP protein, and the number of cells accumulating this protein have been quantitatively determined as a function of time in culture. Parallel determinations have been made for the first two parameters for developing rat brain. The developmental profile of CNP enzymatic activity and amount of CNP protein in culture paralleled that observed in rat brain, in which the period of most active development occurred 7-25 days after birth. Mean CNP activities of 5.6 and 8.1 mumol/min/mg total protein were recorded for the cultures and rat brain, respectively, at their maximal levels. The corresponding mean values for the CNP protein accumulation were calculated to be 138 and 150 pmol/mg total protein, respectively. Thus maximal specific activities of the CNP protein were estimated to be about 800 and 1,000 mumol/min/mg CNP protein for culture and rat brain enzyme, respectively. Approximately three million cells expressing CNP appeared in the cultures per dissociated fetal rat brain seeded. Each CNP+ oligodendrocyte in culture had an average CNP activity of 3.2 pmol/min, and an average CNP protein content of 0.09 fmol (5.4 X 10(7) molecules), values which remained nearly constant during the course of development. Two principal conclusions are drawn from these data. First, the dissociated fetal brain culture system reproduces rather accurately the temporal developmental pattern of CNP expression occurring in the rat brain, but some important quantitative differences occur which suggest the need for additional environmental stimuli missing in these cultures. Second, the quantitative increases in CNP specific activity and amount of CNP protein occurring during oligodendrocyte differentiation in these cultures are primarily the result of increases in the number of CNP+ cells present which upon differentiation express very quickly, via an off-on regulation, steady-state levels of the enzyme. PMID- 2991546 TI - Effects of tunicamycin on NGF binding and neurite outgrowth in PC12 cells. AB - The effects of inhibition of glycosylation on nerve growth factor (NGF) binding and neurite outgrowth response of PC12 cells have been examined. Exposure to tunicamycin (1-10 micrograms/ml) for 24-36 hr eliminates the rapidly dissociating component of NGF binding and decreases the proportion of PC12 cells capable of elaborating neurites in a dose-dependent manner. These decreased cellular responses are probably due to an underglycosylation of the NGF receptor, since the effects of tunicamycin are correlated with a decrease in 3H-fucose incorporation rather than a general decline in cellular metabolism as measured by viability and protein synthesis. These results suggest that carbohydrate side chains are important for the function and/or orientation of the NGF receptor in PC12 cells and that the rapidly dissociating component of NGF binding may be associated with a minimum concentration of functional receptors per cell required for the full biologic response. PMID- 2991547 TI - Cell to cell association in Zajdela ascitic hepatoma. An ultrastructural study. AB - Zajdela ascitic hepatoma cells exist as small islands of two or more cells in addition to single cells. It is now shown that the number of cells per island is directly proportional to the age of the tumor. The number of single cells is maximal 24 h after transplantation of the tumor. Various types of inter-cellular connections are seen between adjacent cells in the island. These junctions are predominant in the apical regions of the cells. The electron-dense material present at junctional sites disappears from several sites after treatment of the islands with chelating agents such as EDTA, EGTA and citrate; the latter being most effective. No appreciable effect on the removal of electron-dense material from the junctional sites is observed when the islands are treated with either collagenase or trypsin. PMID- 2991549 TI - Morphology of nasopharyngeal angiofibroma. An electron microscope study. AB - Three cases of nasopharyngeal angiofibroma were submitted for ultrastructural analysis. All had three main components: cells comprising the microvasculature, fibroblastic (stromal) cells of varying but distinctive morphology, and inflammatory cells. Nuclei of fibroblastic cells that contained dense intranuclear bodies also had a heretofore undescribed prominent fibrous lamina apposed to the inner nuclear envelope. Fibroblastic cells not having intranuclear dense granules also did not have a prominent dense, fibrous lamina (DFL). Thus there appear to be two populations of fibroblastic cells, separable by distinctive nuclear morphology, which together comprise the stromal element of this tumor. The use of Osmium Ferrocyanide as a secondary fixation agent aids considerably in visualization of the intranuclear DFL. This fixation procedure is compared to results obtained using more conventional fixation protocols used in diagnostic electron microscopy. PMID- 2991548 TI - Oncocytic carcinoid of the lung. AB - A case of oncocytic carcinoid of the lung was studied with the electron microscope. This rare tumour is composed of cells acceptable as neuroendocrine cells, oncocytes and cells of an intermediate morphology. Our studies lead us to believe that this tumour develops by oncocytic transformation of neuroendocrine cells in a carcinoid. Two new findings reported in this study are: 1) the occurrence of filamentous inclusions in mitochondria; and 2) calcified spherules which appear to develop in material secreted by tumour cells. PMID- 2991550 TI - Mitochondrial abnormalities in cortical dendrites from patients with Alzheimer's disease. AB - Apparently normal dendrites from the frontal cortex of 7 patients with Alzheimer's disease showed mitochondria with increased density matrices and paracrystalline inclusions in the intercristal space. Most of the inclusions were formed by parallel laminae, while others presented a polyhedric arrangement. Deposits of electron-dense flocculent material were also seen. These abnormal mitochondria appeared in areas of the cortex where dendritic degeneration was more intense. These results, together with identical observations made in the cortex of patients with subacute sclerosing panencephalitis, where dendritic degeneration is also known to occur, led us to advance that mitochondrial abnormalities might precede dendritic degeneration. PMID- 2991551 TI - Intranuclear helioid inclusions. AB - We have coined the term 'helioid inclusion' to describe a new type of single membrane-bound intranuclear inclusion which contains one or more rounded bodies with radiating filaments (helioid bodies). Helioid bodies were not found in the rough endoplasmic reticulum in the cytoplasm, or in the cytoplasmic matrix, but a helioid body was seen in dilated rough endoplasmic reticulum included in a double membrane-bound intranuclear pseudoinclusion. No firm conclusion regarding the genesis of helioid inclusions has emerged from our studies, but two possibilities exist: 1) that helioid bodies derive from modification of serous secretory granules sequestrated in an intranuclear inclusion; or 2) that they develop by condensation of the contents of the perinuclear cistern or rough endoplasmic reticulum subsequent to its sequestration in an intranuclear inclusion. PMID- 2991552 TI - The in vitro actions of clenbuterol (NAB-365) on bovine pulmonary vein and artery. PMID- 2991553 TI - KBSH parvovirus: comparison with porcine parvovirus. AB - We compared the molecular, antigenic, and pathogenic properties of KBSH parvovirus to those of porcine parvovirus (PPV) isolate NADL-8. KBSH, propagated in swine testes cells in culture, possessed two major capsid polypeptides of 83 and 64 kilodaltons that were similar in size to those of PPV. KBSH-infected cells also contained an 86-kilodalton nonstructural polypeptide that was identical in size to the PPV nonstructural polypeptide (NS-1). The KBSH polypeptides were structurally similar but not identical to the corresponding PPV polypeptides, as revealed by partial proteolysis mapping. Viral replicative-form DNA from KBSH infected cells was similar in size to PPV replicative-form DNA and exhibited similar but not identical restriction endonuclease cleavage patterns to that of PPV replicative-form DNA. Antigenically, the two viruses were also very closely related. By using heterologous and homologous antisera, the two viruses were indistinguishable in hemagglutination inhibition and immunoprecipitation assays. However, pathogenically these viruses were dramatically different. NADL-8 caused fetal death when injected into swine fetuses in utero and viremia and high persisting antibody titers when administered orally to weaning-age swine. KBSH inoculated fetuses were normal in appearance, and pigs orally exposed to KBSH failed to establish viremia and demonstrated only transient antibody titers. Thus, KBSH appears to be a PPV that is very closely related to a highly pathogenic PPV isolate, yet is itself nonpathogenic in swine. This reduced pathogenic potential of KBSH may be attributable to its poor ability to replicate in swine. PMID- 2991555 TI - Analysis of wild-derived mice for Fv-1 and Fv-2 murine leukemia virus restriction loci: a novel wild mouse Fv-1 allele responsible for lack of host range restriction. AB - Wild-derived mice originally obtained from Asia, Africa, North America, and Europe were typed for in vitro sensitivity to ecotropic murine leukemia viruses and for susceptibility to Friend virus-induced disease. Cell cultures established from some wild mouse populations were generally less sensitive to exogenous virus than were cell cultures from laboratory mice. Wild mice also differed from inbred strains in their in vitro sensitivity to the host range subgroups defined by restriction at the Fv-1 locus. None of the wild mice showed the Fv-1n or Fv-1b restriction patterns characteristic of most inbred strains, several mice resembled the few inbred strains carrying Fv-1nr, and most differed from laboratory mice in that they did not restrict either N- or B-tropic murine leukemia viruses. Analysis of genetic crosses of Mus spretus and Mus musculus praetextus demonstrated that the nonrestrictive phenotype is controlled by a novel allele at the Fv-1 locus, designated Fv-10. The wild mice were also tested for sensitivity to Friend virus complex-induced erythroblastosis to type for Fv 2. Only M. spretus was resistant to virus-induced splenomegaly and did not restrict replication of Friend virus helper murine leukemia virus. Genetic studies confirmed that this mouse carries the resistance allele at Fv-2. PMID- 2991554 TI - Interstitial murine cytomegalovirus pneumonia after irradiation: characterization of cells that limit viral replication during established infection of the lungs. AB - Interstitial pneumonia associated with viral replication in lung tissue was observed after cytomegalovirus infection of total-body gamma-irradiated mice, whereas in noncompromised hosts the lungs were not affected and virus multiplication was restricted to the salivary glands. The radiation damage could either predispose normally nonpermissive cell types for productive infection or abrogate an immune control of the tissue manifestation of infection by elimination of lymphocytes. Adoptive transfer of lymphoid cells into irradiated, infected recipients supported the second alternative. Even when infection was established in the lungs, as manifested by the presence of infected lung tissue cells in the alveolar septa, an antiviral effect could be assigned to the Lyt-2+, L3T4- subset of T lymphocytes specifically sensitized in the immunocompetent donor. These cells did not require in vitro propagation to perform effector cell functions in vivo and were operative under physiological conditions in comparatively low numbers. Hence, there is reason to assume that T lymphocytes are responsible for the tissue distribution of cytomegalovirus replication during infection. PMID- 2991556 TI - Transformation by Epstein-Barr virus requires DNA sequences in the region of BamHI fragments Y and H. AB - Eight independent recombinant Epstein-Barr virus genomes, each of which was a transforming strain, were made by superinfecting cell lines containing Epstein Barr virus DNA (Raji or B95-8 strain) with a nontransforming virus (P3HR1 strain). A knowledge of the constitution of each transforming recombinant allowed the localization of the defect in the genome of the nontransforming parent to a 12-megadalton sequence within the EcoRI A fragment. Within this region, the nontransforming virus has a deletion of the BamHI Y fragment and about half of the sequences in the adjacent BamHI H fragment. The present data suggest that this deletion is responsible for the nontransforming phenotype. Furthermore, mapping a deletion in one of the recombinant genomes allowed the conclusion that a sequence (comprising about 20% of the Epstein-Barr virus genome) from the center of BamHI-D to BamHI-I' is not necessary for the maintenance of transformation by Epstein-Barr virus. PMID- 2991557 TI - Rous sarcoma virus variants that encode src proteins with an altered carboxy terminus are defective for cellular transformation. AB - The src gene of Rous sarcoma virus (v-src) and its cellular homolog, the c-src gene, share extensive sequence homology. The most notable differences between these genes reside in the region encoding the carboxy terminus of the src proteins. We constructed mutations within the 3' end of the v-src gene to determine the significance of this region to the transforming potential of the v src protein, pp60v-src. The mutants CHdl300 and CHis1511 contain mutations that alter the last 23 amino acids of pp60v-src, whereas the mutant CHis1545-C contains a linker insertion that alters the last 11 amino acids of pp60v-src, and the mutant CHis1545-H contains a linker insertion that results in a 9-amino-acid insertion at position 415. Plasmids bearing each of these mutations were unable to transform chicken cells when introduced into these cells by DNA transfection. In addition, the structurally altered src proteins encoded by the mutants had much-reduced levels of tyrosine protein kinase activity in vivo, as measured by autophosphorylation and phosphorylation of the 34,000-Mr cellular protein, and in vitro, as determined by measuring the level of pp60src autophosphorylation. These data indicate that the carboxy-terminal amino acid sequences play an important role in maintaining the structure of the catalytic domain of pp60v-src. In contrast, the transfection of chicken cells with plasmid DNA containing a chimeric v-c-src gene resulted in morphological cell transformation and the synthesis of an enzymatically active hybrid protein. Therefore, the carboxy terminal sequence alterations observed in the c-src protein do not alone serve to alter the functional activity of a hybrid v-c-src protein appreciably. PMID- 2991558 TI - Thymidine kinase-deficient herpes simplex virus type 2 genital infection in guinea pigs. AB - In guinea pigs, thymidine kinase-producing strains of herpes simplex virus type 2 replicated to high titer in the vagina and spinal cord, and animals developed severe clinical disease. Infection with thymidine kinase-deficient virus resulted in similar vaginal virus titers; however, animals exhibited little or no clinical illness and only low titers of virus were detected in spinal cord homogenate cultures. Neural and extraneural latent infection as well as recurrent infection were noted in animals inoculated with either thymidine kinase-producing or deficient viruses. These data suggest that neural pathways are important in the pathogenesis of genital herpes and that virus-coded thymidine kinase may influence virulence but is not required for latency. PMID- 2991559 TI - Isolation of herpes simplex virus regulatory protein ICP4 as a homodimeric complex. AB - The viral polypeptide ICP4 (or Vmw175) is synthesized during the immediate early phase of infection by herpes simplex virus and regulates the transcription of delayed early and late viral genes. We obtained a partially purified preparation of soluble ICP4 under nondenaturing conditions. Physical constants for native ICP4 were empirically determined by molecular sieve chromatography and sucrose density gradient ultracentrifugation. The Stokes radius of native ICP4 was 8.72 X 10(-7) cm. The sedimentation coefficient of native ICP4 was 9.00S. From these values, the calculated molecular weight of native ICP4 was 342,000, a value which is twice that of monomeric ICP4, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The failure of any other polypeptides to specifically coprecipitate with native ICP4 in the presence of anti-ICP4 antibody indicates that the 342,000-dalton complex is a homodimer of ICP4. The frictional coefficient ratio of native ICP4, which is 1.9, indicates that the homodimer is a highly elongated molecule. PMID- 2991560 TI - Herpes simplex virus 1 mutant deleted in the alpha 22 gene: growth and gene expression in permissive and restrictive cells and establishment of latency in mice. AB - R325-beta TK+, a herpes simplex virus 1 mutant carrying a 500-base-pair deletion in the alpha 22 gene and the wild-type (beta) thymidine kinase (TK) gene, was previously shown to grow efficiently in HEp-2 and Vero cell lines. We report that in rodent cell lines exemplified by the Rat-1 line, plating efficiency was reduced and growth was multiplicity dependent. A similar multiplicity dependence for growth and lack of virus spread at low multiplicity was seen in resting, confluent human embryonic lung (HEL) cells. The shutoff of synthesis of beta proteins was delayed and the duration of synthesis of gamma proteins was extended in R325-beta TK+-infected HEL cells relative to cells infected with the wild-type parent, but no significant differences were seen in the total accumulation of viral DNA. To quantify the effect on late (gamma 2) gene expression, a recombinant carrying the deletion in the alpha 22 gene and a gamma 2-TK gene (R325-gamma 2 TK) was constructed and compared with a wild-type virus (R3112) carrying a chimeric gamma 2-TK gene. In Vero cells, the gamma 2-TK gene of R325 gamma 2TK was expressed earlier than and at the same level as the gamma 2-TK gene of R3112. In the confluent resting HEL cells, the expression of the gamma 2-TK gene of the alpha 22- virus was grossly reduced relative to that of the alpha 22+ virus. Electron microscopic studies indicated that the number of intranuclear capsids of R325-beta TK+ virus was reduced relative to that of the parent virus in resting confluent HEL cells, but the number of DNA-containing capsids was higher. Notwithstanding the grossly reduced neurovirulence on intracerebral inoculation in mice, R325-beta TK+ virus was able to establish latency in mice. We conclude that (i) the alpha 22 gene affects late (gamma 2) gene expression, and (ii) a host cell factor complements that function of the alpha 22 gene to a greater extent in HEp-2 and Vero cells than in confluent, resting HEL cells. PMID- 2991562 TI - Defective interfering virus particles modulate virulence. AB - To determine whether defective interfering (DI) particles modulate virulence by initiating a cyclic pattern of virus growth in vivo, adult mice were infected with vesicular stomatitis virus (VSV), both with and without DI particles. A total of 184 mice divided into groups were inoculated intranasally. A majority of mice inoculated only with standard VSV developed paralysis, most of them between days 7 and 9. The addition of DI particles altered the development of paralysis in several ways. When there was significant protection, a few still became paralyzed on days 7 and 9. When overall mortality was unaffected or even slightly increased, the majority of mice became paralyzed between days 7 and 9 as well. Protection could not be predicted based on a single ratio of standard VSV to DI particles or on the absolute amount of DI particles inoculated. Infectious virus recovered from mouse brains at the time of paralysis and incipient death showed considerable variation, although the titer in a majority of the animals was between 10(5) and 10(7) PFU/ml. When the brains of these paralyzed mice were examined for hybridizable VSV RNA, the detection of standard VSV RNA correlated well with infectivity. The amount of DI RNA in the coinfected mice was more variable and independent of the amount of 40S RNA, although DI RNA was usually found when standard RNA was present. Survivors examined between days 14 and 21 did not contain infectious virus or any detectable viral RNA in their brains. Because these results were consistent with the hypothesis of viral cycling in vivo, rather than a gradual accumulation of total infectious virus, mice were coinfected with 10(8) PFU of standard VSV and 10(5) PFU equivalents of DI particles and sacrificed daily thereafter, irrespective of whether they developed paralysis. Infectivity measurements indicated a reproducible cycling pattern of VSV in the mouse brains with a periodicity of about 5 days. This cycling and the detection of DI RNA in brains several days after intranasal inoculation suggest that there is a dynamic continuous interaction between standard VSV and its DI particle beyond the initial site of replication as the virus population spreads into the host animal. Such cycling of virus production before the full development of specific immune responses from the host may have important implications for viral diagnostics and disease transmission. PMID- 2991561 TI - Identification of a herpes simplex virus function that represses late gene expression from parental viral genomes. AB - The expression of herpes simplex virus gamma 2 (late) genes is inhibited before the onset of viral DNA replication. We report that the block in the expression of certain gamma 2 genes is relieved, at least in part, by defects in the beta ICP8 protein. We have examined the expression of the gamma 2 gene encoding glycoprotein C (gC) in cells infected with a temperature-sensitive ICP8 mutant. Under conditions in which viral DNA replication is inhibited, cells infected with the ICP8 mutant overproduce the gC family of mRNAs relative to the level observed in cells infected with a wild-type virus. The gC mRNA synthesized in cells infected with the ICP8 mutant virus is correctly initiated and spliced and is translated with the same relative efficiency as in cells infected with a replicating wild-type virus. These results suggest that ICP8 is involved in the negative regulation of gamma 2 genes expressed from parental viral genomes. The level of gC expression was greatest in cells infected with a replicating wild type virus. These data suggest that DNA replication and genome amplification are not absolute requirements for gamma 2 gene expression but may facilitate full level expression of these genes. PMID- 2991563 TI - Nucleotide sequence of the Syrian hamster intracisternal A-particle gene: close evolutionary relationship of type A particle gene to types B and D oncovirus genes. AB - We determined the complete nucleotide sequence of the intracisternal A-particle gene, IAP-H18, cloned from the normal Syrian hamster liver DNA. IAP-H18 was 7,951 base pairs in length with two identical long terminal repeats of 376 base pairs at both ends. On the coding strand, imperfect open reading frames corresponding to gag and pol of the retrovirus genome were observed, whereas many stop codons were present in the region corresponding to env. The putative H18 gag gene (809 amino acids) had a sequence homologous to the N-terminal half of the mouse mammary tumor virus gag gene and locally to the Rous sarcoma virus gag gene. The putative H18 pol gene (900 residues) was homologous to the Rous sarcoma virus pol gene almost throughout the entire region. Two conserved regions among the retrovirus pol genes have been reported. One presumably corresponds to the DNA polymerase and the RNase H domain, and the other corresponds to the DNA endonuclease domain of the multifunctional protein pol. By the comparison of the deduced amino acid sequences of the putative endonuclease domain of six representative oncovirus genomes, a phylogenetic tree of the oncovirus genomes was constructed, and the intracisternal A-particle (type A) genome was found to be more closely related to the mouse mammary tumor virus (type B) and squirrel monkey retrovirus (type D) genomes. PMID- 2991564 TI - Isolation and immunizations with hepatitis A viral structural proteins: induction of antiprotein, antiviral, and neutralizing responses. AB - An immune affinity purification procedure for hepatitis A virus (HAV) was designed which yielded milligram quantities of the virus with greater than 95% purity. The major structural proteins VP-1, VP-2, and VP-3 were isolated from the purified virus by electroelution from sodium dodecyl sulfate-polyacrylamide gels and used to immunize Lewis rats (three to four doses, 10 to 15 micrograms per dose). The two Lewis rats immunized with VP-1 developed a strong antibody response to VP-1, as determined by Western blot analysis and immune precipitation of the denatured protein. These animals also developed a good antibody response to the whole virus, as demonstrated by a positive response in a competitive radioimmunoassay (HAV antibody test) and by precipitation of the whole virus. In addition, both animals developed a low titer neutralizing antibody to HAV, as demonstrated by an in vitro cell culture assay. While the two rats receiving VP-2 developed only minimal responses to the protein and to the virus by the same assays described above, one of the two developed a significant neutralizing antibody to HAV. The immunization of one Lewis rat with VP-3 induced a good antibody response to both denatured protein and the whole virus. This serum sample was also demonstrated to neutralize the viral infectivity. Finally, two rabbits that had received inoculations of sodium dodecyl sulfate and heat disrupted HAV (containing 20 to 30 micrograms of each protein per dose) developed good antiprotein responses to all of the proteins and good antiviral responses, including a consistently significant neutralizing activity. The neutralizing antibody responses suggest that the structural proteins of HAV, or a portion of them, could provide the basis for a subunit vaccine for HAV. PMID- 2991565 TI - The placenta as a site of cytomegalovirus infection in guinea pigs. AB - The development of cytomegalovirus (CMV) infection in the placenta was studied in Hartley guinea pigs inoculated at midgestation, and its role in determining the outcome of fetal CMV infection was assessed. A hematogenous spread of CMV from the mother to the placenta occurred early during the course of the infection. However, the virus remained present in placental tissues long after CMV had been cleared from maternal blood (i.e., 3 and 4 weeks postinoculation). At that time, the virus was able to replicate in placental tissues in the presence of specific maternal antibodies. Viral nucleocapsids were seen within nuclei of trophoblastic cells, and virions were present surrounding infected cells. In addition, typical CMV-induced histopathological lesions bearing CMV antigens were consistently localized at the transitional zone between the capillarized labyrinth and the noncapillarized interlobium. Whenever CMV infection of the fetus occurred, virus was isolated from the associated placenta. Among placental-fetal units with CMV infected placentas, only 27% of the fetuses were found to be infected. In addition, there was a delay in the establishment of the infection in the fetus in relation to the placenta, although frequencies of virus isolation in placental and fetal tissues peaked at 3 weeks after CMV inoculation. These results suggest that during primary CMV infection of pregnant guinea pigs, the placenta not only serves as a reservoir for CMV but also acts to limit transmission of the virus to the fetus. PMID- 2991566 TI - Establishment of latency in mice by herpes simplex virus 1 recombinants that carry insertions affecting regulation of the thymidine kinase gene. AB - Herpes simplex virus 1 recombinants carrying alpha-, beta-, and late gamma (gamma 2)-regulated thymidine kinase (TK) genes were tested for the ability to establish latency in BALB/c mice inoculated by the eye route. The significant findings were as follows. Representatives of alpha- and gamma 2-regulated TK recombinants all established and maintained latent infections, but the efficiency was somewhat lower than that of wild-type virus. Of the three alpha TK recombinants tested, one (R316) spontaneously deleted portions of the inserted sequences which conferred alpha regulation to the TK gene. The viruses carrying these deletions expressed considerably lower TK activity than did wild-type virus, i.e., 2 to 40% of the levels expressed by the wild-type virus carrying the beta TK gene. However, the ability of these viruses to establish latency was not related to the efficiency of expression of the TK gene. These results indicate the following: (i) conversion of the TK gene into an alpha or gamma 2 gene did not preclude the establishment of latent infections; (ii) there was no correlation between the levels of TK activity expressed in cell culture and the ability to establish latency; and (iii) rearrangement of the genome by insertions or deletions which interrupt gene domains did not automatically result in an inability to establish latent infections. PMID- 2991567 TI - Activation of the major immediate early gene of human cytomegalovirus by cis acting elements in the promoter-regulatory sequence and by virus-specific trans acting components. AB - Upstream of the major immediate early gene of human cytomegalovirus (Towne) is a strong promoter-regulatory region that promotes the synthesis of 1.95-kilobase mRNA (D. R. Thomsen, R. M. Stenberg, W. F. Goins, and M. F. Stinski, Proc. Natl. Acad. Sci. U.S.A. 81:659-663, 1984; M. F. Stinski, D. R. Thomsen, R. M. Stenberg, and L. C. Goldstein, J. Virol. 46:1-14, 1983). The wild-type promoter-regulatory region as well as deletions within this region were ligated upstream of the thymidine kinase, chloramphenicol acetyltransferase, or ovalbumin genes. These gene chimeras were constructed to investigate the role of the regulatory sequences in enhancing downstream expression. The regulatory region extends to approximately 465 nucleotides upstream of the cap site for the initiation of transcription. The extent and type of regulatory sequences upstream of the promoter influences the level of in vitro transcription as well as the amount of in vivo expression of the downstream gene. The regulatory elements for cis activation appear to be repeated several times within the regulatory region. A direct correlation was established between the distribution of the 19 (5' CCCCAGTTGACGTCAATGGG 3')- and 18 (5' CACTAACGGGACTTTCCAA 3')-nucleotide repeats and the level of downstream expression. In contrast, the 16 (5' CTTGGCAGTACATCAA 3')-nucleotide repeat is not necessary for the enhancement of downstream expression. In a domain associated with the 19- or 18-nucleotide repeats are elements that can be activated in trans by a human cytomegalovirus-specified component but not a herpes simplex virus-specified component. Therefore, the regulatory sequences of the major immediate early gene of human cytomegalovirus have an important role in interacting with cellular and virus-specific factors of the transcription complex to enhance downstream expression of this critical viral gene. PMID- 2991568 TI - Transcriptional and translational analysis of the vaccinia virus late gene L65. AB - Among the products of vaccinia virus genes which are expressed late in infection is a major polypeptide (Mr, 65,000) designated L65. Pulse-chase analyses indicated that L65 is not subject to posttranslational cleavage as is the core polypeptide p4b which migrates to a similar position in sodium dodecyl sulfate polyacrylamide gels. A polypeptide of 65,000 molecular weight produced in reticulocyte lysates programmed with viral mRNA isolated late in infection was identified as L65 by peptide mapping. L65 mRNA was purified by hybridization selection to restriction fragments of the viral genome and translated in vitro. This allowed the gene encoding L65 to be mapped to the rightmost 4.5 kilobase pairs of the HindIII D fragment. Transcriptional mapping of this region of the genome detected a late mRNA which was initiated at 450 base pairs to the right of the HindIII D-A junction, was transcribed in the leftward direction, and was terminated in the nondescript manner typical of vaccinia virus late mRNAs. PMID- 2991569 TI - DNA packaging initiation of Salmonella bacteriophage P22: determination of cut sites within the DNA sequence coding for gene 3. AB - DNA packaging of Salmonella phage P22 starts at a defined site on a concatemer of P22 genomes. The molecular ends formed at the packaging initiation site (pac) map within a region of ca. 120 base pairs and may contain any of the four nucleotides at their 5' end. The determination of the positions of the cuts within the sequence demonstrates a characteristic distribution of cut sites which apparently cannot be attributed to the sequence organization of the involved regions. Symmetric elements of the sequence might serve as signals for a recognition event(s) at pac in a separate process preceding the cutting reaction. The region of packaging initiation is located within the sequence coding for gene 3. The 3 protein is responsible for the site specificity of this process. We find no significant homology to Nu1 protein, which appears to have an analogous or similar function in the DNA maturation of Escherichia coli phage lambda. PMID- 2991570 TI - Specificities of monoclonal and polyclonal antibodies that inhibit adsorption of herpes simplex virus to cells and lack of inhibition by potent neutralizing antibodies. AB - Polyclonal and monoclonal antibodies to individual herpes simplex virus (HSV) glycoproteins were tested for ability to inhibit adsorption of radiolabeled HSV type 1 (HSV-1) strain HFEMsyn [HSV-1(HFEM)syn] to HEp-2 cell monolayers. Polyclonal rabbit antibodies specific for glycoprotein D (gD) or gC and three monoclonal mouse antibodies specific for gD-1 or gC-1 most effectively inhibited HSV-1 adsorption. Antibodies of other specificities had less or no inhibitory activity despite demonstrable binding of the antibodies to virions. Nonimmune rabbit immunoglobulin G and Fc fragments partially inhibited adsorption when used at relatively high concentrations. These results suggest involvement of gD, gC, and perhaps gE (the Fc-binding glycoprotein) in adsorption. The monoclonal anti gD antibodies that were most effective at inhibiting HSV-1 adsorption had only weak neutralizing activity. The most potent anti-gD neutralizing antibodies had little effect on adsorption at concentrations significantly higher than those required for neutralization. This suggests that, although some anti-gD antibodies can neutralize virus by blocking adsorption, a more important mechanism of neutralization by anti-gD antibodies may be interference with a step subsequent to adsorption, possibly penetration. PMID- 2991571 TI - Potent neutralizing activity associated with anti-glycoprotein D specificity among monoclonal antibodies selected for binding to herpes simplex virions. AB - Thirty-three monoclonal antibodies were selected for ability to bind to purified virions of herpes simplex virus and were shown by immunoprecipitation to react with one or another of the envelope glycoproteins. Only six of these antibodies exhibited potent neutralizing activity, and all six were specific for glycoprotein D. Two other anti-glycoprotein D antibodies and 25 antibodies specific for four other viral glycoproteins had much less potent, if any, neutralizing activity. PMID- 2991572 TI - Poliovirus protease 3C (P3-7c) does not cleave P220 of the eucaryotic mRNA cap binding protein complex. AB - Infection of HeLa cells by poliovirus results in proteolysis of the large subunit (P220) of the cap-binding protein complex. This is believed to cause the rapid shut-off of host protein synthesis during poliovirus infection. In this communication we examined the possible involvement of poliovirus proteins 3C (a proteinase) and 2C in cleavage of P220. Using antisera against these two viral polypeptides, we were unable to inhibit proteolysis of P220 in an in vitro assay. These results indicate that viral proteins 3C and 2C are not directly involved in cleaving P220 and hence do not cause shut-off of cellular protein synthesis. PMID- 2991573 TI - Expression of the x-lor gene of human T-cell leukemia virus I in Escherichia coli. AB - The 3'-terminal regions of the human T-cell leukemia virus I (HTLV-I) and HTLV-II genomes encode a novel gene product. We showed that expression of this region fused to the beta-galactosidase gene in bacteria produces a protein recognized by adult T-cell leukemia-lymphoma patient sera. Rabbit antibodies raised against this protein specifically precipitated the 42-kilodalton x-lor gene protein from HTLV-I-infected cells. PMID- 2991574 TI - Early clonality and high-frequency proviral integration into the c-myc locus in AKR leukemias. AB - Blot hybridization of thymocyte DNA from AKR/J mice was used to detect new proviral junction fragments as markers of clonality at different stages of viral leukemogenesis and to detect DNA rearrangements at the c-myc locus due to proviral insertion. Clonal populations of thymocytes were observed in mink cell focus-forming virus-injected mice as early as 35 days postinjection, at a stage distinguishable from frank leukemia by flow cytometric analysis and transplantation bioassay. Specific proviral integrations in the c-myc locus were detected in 15% of these early clones and in up to 65% of late-developing thymomas and frank leukemias. Thus, in this system c-myc activation appears to be a common mechanism in T-cell leukemogenesis. PMID- 2991575 TI - Rescue of a herpes simplex virus type 1 neurovirulence function with a cloned DNA fragment. AB - A herpes simplex virus type 1 (HSV-1) genetic function that is required for viral replication in the murine central nervous system was unambiguously localized. Thus, cosmid clones of either HSV-1 HindIII fragment C (0.64 to 0.87 map units) or fragment B (0.64 to 0.83 plus 0.91 to 1.0 map units) were employed to restore neurovirulence to an intertypic recombinant (RE6) that is specifically deficient in this property. The neurovirulent recombinants were generated in cell culture by cotransfecting the clone fragments and unit-length RE6 DNA and then selected in mouse brains. Either fragment efficiently conferred neurovirulence to RE6, demonstrating that no short region unique sequences are required. Analyses of the genomic structures of the neurovirulent recombinants showed that, in every case, HSV-1 information from 0.71 to 0.83 map units was incorporated into the RE6 genome. Cleavage of HindIII fragment C with EcoRI eliminated its capacity to rescue RE6. Virulence could be restored by the addition of HSV-1 BamHI fragment L (0.71 to 0.74 map units) that spans an EcoRI site at 0.72 map units. The precise location of this HSV-1 neurovirulence function is discussed. PMID- 2991576 TI - Diagnosis of rotavirus infection with cloned cDNA copies of viral genome segments. AB - The diagnostic potential of cloned cDNA copies of human rotavirus (strain WA) genome segments for the detection of rotavirus in clinical specimens has been determined. A hybridization assay in which a mixture of 32P-labeled cDNAs representing the 11 rotavirus segments was used as a probe compared favorably with three frequently used diagnostic tests for rotavirus in terms of both specificity and sensitivity. Significantly, clinical isolates could be readily distinguished when cloned cDNA copies of individual genome segments were used independently as a probe. In assays in which genome RNA from rotaviruses of known subgroups and serotypes were tested, cloned probes that encode nonstructural viral proteins hybridized efficiently to genome RNAs of all strains, whereas cloned probes corresponding to genome segments 6 and 9 exhibited the potential for differentiating strains of different subgroups and serotypes. Cloned cDNA copies of rotavirus genome segments therefore offer considerable potential for improved general diagnosis of rotavirus in clinical specimens, as well as for epidemiological studies in which virus isolates can be distinguished on the basis of nucleotide sequence homology of individual genome segments. PMID- 2991577 TI - Deletion of the gag region from FBR murine osteosarcoma virus does not affect its enhanced transforming activity. AB - Both FBJ murine osteosarcoma virus (FBJ-MSV) and FBR-MSV induce transformation in tissue culture and osteogenic sarcomas in mice. In tissue culture, however, FBR MSV induces larger foci with a shorter latency than those induced by FBJ-MSV. Transformation is dependent on expression of the fos oncogene in FBJ-MSV and of a gag-fos fusion protein in FBR-MSV. We have determined that the gag sequences can be deleted from FBR-MSV without affecting the high transforming activity of this virus in comparison to FBJ-MSV. The resultant virus, designated FBJ/R-MSV, has a protein coding region that is half the size of that of FBR-MSV and about one third smaller than that of FBJ-MSV. Thus, FBJ/R-MSV will provide a useful tool for studying the transforming activity of the fos oncogene. PMID- 2991578 TI - Monocyte-derived inhibitor of interleukin 1 induced by human cytomegalovirus. AB - It has previously been shown that human cytomegalovirus (HCMV) can exert immunosuppressive effects, and it has been suggested that these may be mediated by monocytes, although the mechanism is unclear. We showed that infection of human monocytes with the AD169 strain of HCMV abrogates their production of interleukin 1 (IL-1) activity. This was associated with the release from infected monocytes of an inhibitor of IL-1 activity which was also released after HCMV infection of the U937 macrophage-like cell line. The inhibitor of IL-1 activity is a protein with an apparent molecular weight of ca. 95,000. This action of HCMV strain AD169 was virus specific and required infectious virus but occurred without virus replication or detectable expression of viral proteins. This effect may account, at least in part, for the previously observed immunosuppressive properties of HCMV. PMID- 2991580 TI - Purification of a HeLa cell receptor protein for group B coxsackieviruses. AB - Coxsackievirus B3 (CB3) firmly attaches to HeLa cells, forming a specific complex between the virus and its receptor on the cell surface. We extracted this virus receptor complex (VRC) with the detergents sodium deoxycholate and Triton X-100. The VRC was identified by its sedimentation coefficient (140S), which was less than that of virions (155S). Formation of the VRC from cell lysates and 3H-CB3 occurred with the same specificity as did attachment of virions to cells, in that its formation was blocked by unlabeled CB3 but not by poliovirus. The VRC was purified 30,000-fold by differential and sucrose gradient centrifugation. Iodination with Na125I revealed that the purified VRC consisted of the normal CB3 proteins and one additional protein (RP-a) with an approximate molecular weight of 49,500. RP-a was eluted from the VRC and was shown to rebind with CB3 and CB1 virions but not with poliovirus type 1. We propose that Rp-a is a protein in the plasma membrane receptor complex which is responsible for the specific recognition and binding of the group B coxsackieviruses. PMID- 2991579 TI - Different genes control the susceptibility of mice to Moloney or Abelson murine leukemia viruses. AB - The susceptibility of mice to lymphoma induction by Moloney or Abelson murine leukemia virus has been compared in BALB/c, C57BL/6, and BALB/cXC57BL/6 recombinant inbred strains. BALB/c mice were found to be susceptible to lymphoma induction by either virus, and C57BL/6 mice were found to be relatively resistant to lymphoma induction by either virus. The genes that control these patterns of susceptibility to each virus are not the same because susceptibility to each virus segregated independently in CXB recombinant inbred strains. We also found, as reported by Cook (W. Cook, Proc. Natl. Acad. Sci. U.S.A. 79:2917-2921, 1982), when injected intrathymically that Abelson murine leukemia virus rapidly induced thymomas in weanling B6 mice. Examination of the cellular phenotypes of the tumors induced by Abelson murine leukemia virus or by Moloney murine leukemia virus indicated that different lymphocyte subpopulations were the targets for tumor induction by each virus. PMID- 2991581 TI - Cloning and sequence of DNA encoding structural proteins of the autonomous parvovirus feline panleukopenia virus. AB - Approximately 80% of the genome of feline panleukopenia virus was cloned into pBR322. This DNA included the transcription unit for the major viral mRNA species. The nucleotide sequence of the cloned portion of the genome was determined. Comparison of the feline panleukopenia virus sequence with the sequences of the parvoviruses minute virus of mice and H-1 revealed considerable homology between the three viruses on both the nucleic acid and protein levels. Based on this homology, a model for the generation of the two size classes of viral structural proteins (VP1 and VP2') is proposed. PMID- 2991582 TI - Neuropeptide-induced hypothermia and the course of central nervous system disease mediated by temperature-sensitive mutants of vesicular stomatitis virus. AB - Mice inoculated with many temperature-sensitive (ts) vesicular stomatitis virus (VSV) mutants incur a less aggressive disease than mice infected with wild-type VSV. The normal body temperature of mice, 38 degrees C, is not a permissive temperature for replication of the temperature-sensitive VSV mutants in cell culture. To determine whether the body temperature of mice caused the alteration in disease states, a neuropeptide that induces hypothermia in rodents was injected into mice before their infection with a temperature-sensitive VSV mutant. Only 1.0 ng of the neuropeptide neurotensin, injected intracerebroventricularly, was required to lower the core temperatures of mice an average of 2.5 degrees C. A single injection of neurotensin before infection with tsG31 VSV (complementation group III) dramatically altered the course of disease. Without neurotensin only 3% of the mice infected with tsG31 VSV died, but when neurotensin was administered 24 h before the inoculation of the tsG31 VSV, 80% of the mice died. The course of disease in mice produced by infection with another temperature-sensitive VSV mutant, tsG11 VSV (complementation group I), also was altered when neurotensin was injected before inoculation of the virus. Instead of 3% of the mice dying as in a normal infection with tsG11 VSV, treatment with neurotensin before inoculation produced a rapidly fatal disease, killing 90% of the mice. PMID- 2991583 TI - Inhibition of viral mRNA translation in interferon-treated L cells infected with reovirus. AB - Murine L cells were treated with interferon (IFN) concentrations which reduced by 75 to 80% the synthesis of viral mRNA after infection with reovirus. Protein synthesis was not inhibited in these cells up to 6 h after infection, but a large fraction of the viral mRNA was not associated with polyribosomes and sedimented at about 50S. In contrast, most of the reovirus mRNA was associated with polyribosomes in control infected cells. This mRNA was of similar size to non polyribosomal mRNA from IFN-treated cells when analyzed by Northern blot hybridization with a cloned cDNA for the s2 reovirus mRNA, indicating that the non-polyribosomal mRNA was not appreciably degraded. Viral mRNA was labeled with [3H]uridine and the non-polyribosomal mRNA was isolated from IFN-treated cells. This mRNA could quantitatively bind to 80S initiation complexes when incubated in a rabbit reticulocyte cell-free system. These findings indicated that the non polyribosomal RNA was translatable, but that its binding to functional initiation complexes was inhibited in IFN-treated cells by a discriminatory mechanism, which did not affect translation of cellular mRNA. Previous experiments showed that mRNA is blocked in 48S complexes when the alpha subunit of initiation factor eIF 2 is phosphorylated by the double-stranded RNA-dependent protein kinase induced by IFN. A localized activation of this kinase could explain the block of viral mRNA in 48S complexes. By labeling the phosphoproteins of IFN-treated cells with 32P, eIF-2 (alpha P) was shown to cosediment with non-polyribosomal mRNA, presumably in 48S complexes. PMID- 2991584 TI - Susceptibility to erbB-induced erythroblastosis is a dominant trait of 151 chickens. AB - Rous-associated virus-1 (RAV-1)-induced erythroblastosis results from proviral insertions into or viral transductions of the c-erbB region of the epidermal growth factor gene. Most chickens develop low incidences (less than 5%) of RAV-1 induced erythroblastosis. However, an inbred line of chickens (151) suffers high incidences (approximately 80%) of RAV-1-induced erythroblastosis. Analysis of 151, K28, and (K28 X 151) X K28 chickens for susceptibility to RAV-1-induced erythroblastosis revealed that susceptibility to RAV-1-induced erythroblastosis is a dominant trait of line 151 chickens. Analysis of 151 X K28 and K28 chicks for susceptibility to the induction of erythroblastosis by two new c-erbB transducing viruses (avian erythroblastosis virus strains AEV-5005 and AEV-5009) revealed that susceptibility to transformation by new c-erbB-transducing viruses is also a dominant trait of 151 chickens. We think it is likely that both of these dominant traits are encoded by the same gene or genes. Our hypothesis is that this gene (or genes) potentiates the ability of the transmembrane and cytoplasmic domains of the epidermal growth factor receptor to transform cells. PMID- 2991585 TI - Herpesvirus saimiri-induced lymphoblastoid rabbit cell line: growth characteristics, virus persistence, and oncogenic properties. AB - A nonproducer lymphoblastoid cell line (7710) containing the herpesvirus saimiri (HVS) genome was established from the HVS-positive spleen of a male, inbred New Zealand White rabbit (III/J strain) which had developed a well-differentiated lymphoma after inoculation of HVS and 12-O-tetradecanoylphorbol-13-acetate (TPA). Antibodies to HVS early and late antigens were detected in the serum of rabbit 7710 by indirect immunofluorescence and immunoprecipitation. The cell line was of T-cell origin, did not produce HVS, and could not be superinfected with HVS. However, HVS early antigens could be induced in the cells with n-butyric acid and TPA or TPA alone. On the other hand, late antigens were never observed, and infectious virus could not be rescued by cocultivation of 7710 cell with OMK cells. The 7710 cells were T-cell growth factor dependent, even after many in vitro passages. The 7710 cell line contained multiple copies of a nonintegrated, covalently closed circular HVS genome. As is characteristic of some other HVS transformed nonproducer lymphoid cell lines, a large segment of unique light (L) DNA was missing in the persistent circular viral DNA present in 7710 cells. This deletion spanned at least 42.5 kilobases, corresponding to the segment between 12.3 and 50.7 map units of full-length, infectious virion L-DNA. The 7710 cells failed to induce tumors in athymic nude mice, but inbred rabbits inoculated with as few as 100 of these cells developed fatal lymphomas. Chromosomal analysis showed that tumors were due to the growth of donor cells. Cells recovered from one of the rabbits inoculated with 7710 cells also contained HVS DNA and, after in vitro culture, induced the same type of lymphoma when inoculated into two other III/J-strain rabbits. None of the previously described HVS-transformed cell lines have been able to induce tumors in either their host species or nude mice. Thus, our demonstration that the 7710 cell line is readily transplantable in syngeneic rabbits represents the first available model which allows analysis of many biological and molecular aspects of the in vivo oncogenicity of HVS. PMID- 2991586 TI - Intracellular location of rabbit poxvirus nucleic acid within infected cells as determined by in situ hybridization. AB - The intracellular location of rabbit poxvirus DNA within cells during the course of infection has been determined by the hybridization in situ of labeled viral DNA probes to uninfected and infected cells under various conditions. Extensive control experiments were performed to demonstrate that DNA could be detected selectively and accurately within the cell. Our results suggest that rabbit poxvirus DNA is located only within the cytoplasm during the reproductive cycle, and we found no evidence that viral DNA enters the cell nucleus. The pattern of hybridization of viral DNA at early times (1 and 2 h postinfection) and in the presence of inhibitors of viral DNA synthesis suggests that there may be an association between the input viral DNA and some structural component of the host cell. A number of observations support the hypothesis that the host cell nucleus is required for a productive poxvirus infection. Our results are discussed in terms of the possible role of the nucleus in the replication of poxviruses. PMID- 2991587 TI - Reduced microfilament organization in adenovirus type 5-infected rat embryo cells: a function of early region 1a. AB - The actin microfilament organization in rat embryo cells was examined by fluorescence microscopy with 7-nitrobenz-2-oxa-1,3-diazole-phallacidin and by electron microscopy, after mock infection or infection with adenovirus type 5 (Ad5). Infected cells showed severely reduced numbers of actin microfilaments and stress fibers, detectable early after infection. Mutants defective in Ad5 early genes were used to show that reduced microfilament organization was a function of the Ad5 transformation early gene 1a (E1a) and did not require expression of any other viral gene. The product of the E1a 13s mRNA was essential for the effect, although the 12s mRNA product appeared to contribute. Ad5 infection of the cells had no observable effect on total cell actin levels or on the ratio of monomeric to polymeric actin. E1a, therefore, affected only the higher-order organization of actin. PMID- 2991588 TI - Construction and biological analysis of deletion mutants of Fujinami sarcoma virus: 5'-fps sequence has a role in the transforming activity. AB - Fujinami sarcoma virus (FSV) genome codes for the gag-fps fusion protein FSV P130. The amino acid sequence of the 3' one-third portion in v-fps is partially homologous to the 3' half of pp60src, or the kinase domain, but the sequence of the 5' portion is unique to v-fps. To identify a possible domain structure in the v-fps sequence responsible for cell transformation, we constructed various deletion mutants of FSV with molecularly cloned viral DNA. Their transforming activities were assayed by measuring focus formation on chicken embryo fibroblasts and rat 3Y1 cells and tumor formation in chickens. The mutants carrying a deletion at the 3' portion in v-fps, the kinase domain, lost transforming activity. The mutants carrying an approximately 1-kilobase deletion within the 5' portion of the v-fps sequence retained focus-forming activity and tumorigenicity in the chicken system, but the efficiency of focus formation was about 10 times lower than that of the wild type. The morphology of these transformed cells was distinct from that observed in cells infected with wild type FSV. Furthermore, these mutants could not transform rat 3Y1 cells, although wild-type FSV DNA transformed rat 3Y1 cells at a high frequency. The mutants carrying a larger deletion in the 5' portion of fps completely lacked the transforming activity. These results suggest that the 3' portion of the v-fps sequence is necessary but not sufficient for cell transformation and that the 5' portion of v-fps has a role in the transforming activity. PMID- 2991589 TI - Isolation and characterization of monoclonal antibodies directed against two subunits of rabbit poxvirus-associated, DNA-directed RNA polymerase. AB - A library of monoclonal antibodies directed against individual proteins of the rabbit poxvirus (RPV) virion within a complex immunogenic mixture has been generated through the use of in vivo and in vitro immunization regimens. The relative efficacies of the two procedures were compared. Based on immunoblot analysis, the in vitro immunization regimen led both to a wider variety of monoclonal antibodies to different proteins and to a larger number of antibodies directed against proteins of higher molecular weights. Each method, however, has advantages, and the two procedures appear to be complementary. A simple method to recognize antibodies directed against the virion DNA-directed RNA polymerase was developed. Monoclonal antibodies directed against two subunits (137 and 34 kilodaltons [kDa]) of the RNA polymerase were identified and used to study the biogenesis of the enzyme and to map the two corresponding genes within the viral genome by using an RPV DNA library cloned into the lambda gtll expression vector. Both proteins are synthesized late in the infectious cycle and are restricted totally to the cytoplasm. Preliminary mapping data place the genes encoding the 137-kDa protein within the HindIII H fragment, whereas the gene for the 34-kDa protein is located within the left most region of the HindIII A fragment. PMID- 2991590 TI - Susceptibility of wild mouse cells to exogenous infection with xenotropic leukemia viruses: control by a single dominant locus on chromosome 1. AB - Although xenotropic murine leukemia viruses cannot productively infect cells of laboratory mice, cells from various wild-derived mice can support replication of these viruses. Although the virus-sensitive wild mice generally lack all or most of the xenotropic proviral genes characteristic of inbred strains, susceptibility to exogenous infection is unrelated to inheritance of these sequences. Instead, susceptibility is controlled by a single dominant gene, designated Sxv, which maps to chromosome 1. Sxv is closely linked to, but distinct from Bxv-1, the major locus for induction of xenotropic murine leukemia viruses in laboratory mice. Genetic experiments designed to characterize Sxv show that this gene also controls sensitivity to a wild mouse virus with the interference properties of mink cell focus-forming murine leukemia viruses, and that Sxv-mediated susceptibility to xenotropic murine leukemia viruses is restricted by the mink cell focus-forming virus resistance gene Rmcf. These data, together with genetic mapping of the mink cell focus-forming virus cell surface receptor locus to this same region of chromosome 1, suggest that Sxv may encode a wild mouse variant of the mink cell focus-forming virus receptor that allows penetration by xenotropic murine leukemia viruses. PMID- 2991592 TI - Correspondence between immunological and functional domains in the transforming protein of Fujinami sarcoma virus. AB - Monoclonal antibodies reactive with either gag or fps portions of the wild-type Fujinami sarcoma virus transforming protein have been used to probe the structure of proteins encoded by mutant genomes constructed in vitro. The pattern of immunoreactivity suggests that the functional domain defined in genetic studies (Stone et al., Cell 37:549-558, 1984) corresponds to a discrete immunological domain in the native, wild-type Fujinami sarcoma virus protein. At least one mutation affecting both the structure and function of the proposed NH2-terminal fps-specific domain encodes a product with high specific activities in kinase assays. Furthermore, a cell line expressing high levels of this mutant protein is only moderately transformed. The striking correspondence between the immunological domain defined here and the functional domain inferred from the results of transfection experiments suggests that this non-kinase-specifying region constitutes a discrete structural as well as functional component of the viral protein. PMID- 2991591 TI - Epstein-Barr virus-encoded protein found in plasma membranes of transformed cells. AB - We have developed monoclonal antibodies to a 63,000-molecular-weight protein (p63) which is the product of the most abundant messenger RNA in Epstein-Barr virus-transformed cells and shown that the protein is associated specifically with plasma membranes. It was also found to be associated with the other membrane fractions and was found in all Epstein-Barr virus-transformed cells tested. In addition, p63 was present in virions, resulting in transient, early appearance in newly infected cells. Newly synthesized p63 was detected at the time cells underwent blast transformation (48 to 72 h postinfection). The possible role of this protein in transformation and as a target for cell-mediated cytotoxicity is discussed. PMID- 2991593 TI - Partial nucleotide sequence of Rous sarcoma virus-29 provides evidence that the original Rous sarcoma virus was replication defective. AB - Rous sarcoma virus-29 (RSV-29) is the strain of RSV that has the least number of passages beyond its isolation from chicken tumor no. 1 among all current strains of RSV. Biological characterization indicated that it was replication defective. RNA analysis of nonproducer clones of RSV-29-infected chicken embryonic fibroblasts showed the presence of a subgenomic message of 2.6 kilobases containing src and a genomic RNA of 7.7 kilobases that contains gag, pol, and src, but not env. The src-containing EcoRI fragment of RSV-29 proviral DNA was molecularly cloned. Sequence analysis of the regions flanking src revealed that the env gene was completely deleted in RSV-29 and that the sequence across the deletion was exactly the same as the Bryan high-titer strain of RSV. The sequence immediately 3' to src in RSV-29 was closely related to that of the Prague strain of RSV. The fact that the strain of RSV which has the minimal number of passages beyond its isolation is replication defective supports the hypothesis of Lerner and Hanafusa (J. Virol. 49:549-556, 1984) that the original RSV is a defective transforming virus. This defective transforming virus is postulated to be the precursor to other defective RSVs like the Bryan high-titer strain and to nondefective RSVs like the Prague strain. The particular clone of RSV-29 that we studied also had a short stretch of sequence duplication at the 3' end of the pol gene, which was presumably created by an error of reverse transcription. PMID- 2991594 TI - Sequences outside of the long terminal repeat determine the lymphomogenic potential of Rous-associated virus type 1. AB - Recombinant avian leukosis viruses have been constructed from the molecularly cloned DNAs of Rous-associated virus type 1 (RAV-1) and Rous-associated virus type 0(RAV-0). Virus encoded by the cloned RAV-1 DNA induced a high incidence of B-cell lymphoma and a moderate incidence of a variety of other neoplasms. Virus encoded by the cloned RAV-0 DNA did not cause disease. Virus recovered from DNA constructions that encoded the gag, pol, and 5' env sequences of RAV-0 and the 3' env and long terminal repeat sequences of RAV-1 did not cause a high incidence of lymphoma. Rather, these constructed viruses induced a low incidence of a variety of neoplasms. Virus recovered from reconstructed pRAV-1 DNA had the same disease potential as did virus recovered from the parental pRAV-1 DNA. These results indicate that the long terminal repeat sequences of RAV-1 do not confer the potential to induce a high incidence of B-cell lymphoma. PMID- 2991595 TI - Characterization of a molecularly cloned retroviral sequence associated with Fv-4 resistance. AB - The murine leukemia virus (MuLV) sequence associated with the resistance allele of the Fv-4 gene (Fv-4r) was molecularly cloned from genomic DNA of uninfected mice carrying this allele. The 5.2-kilobase cloned EcoRI DNA fragment (pFv4) was shown by nucleotide sequencing to contain 3.4 kilobases of a colinear MuLV related proviral sequence which began in the C-terminal end of the pol region and extended through the env region and the 3' long terminal repeat. Cellular sequences flanked the 3' as well as the 5' ends of the truncated MuLV sequence. Alignment of the N-terminal half of the pFv4 env sequence with ecotropic, mink cell focus-forming, and xenotropic MuLV env sequences established the relatedness of pFv4 and ecotropic MuLV env sequences. A subcloned 700-base pair segment (pFv4env) from the 5' env region of pFv4 was used as an Fv-4-specific probe; it hybridized specifically to the Fv-4r-associated proviral sequence but not to endogenous ecotropic MuLV proviral DNA under high stringency. All Fv-4-resistant mice contained the same retroviral segment associated with the same flanking cellular DNA. Expression of Fv-4r-specific mRNA was demonstrated in the spleens of Fv-4r mice but not Fv-4s mice, supporting the previously proposed resistance model based on interference. PMID- 2991596 TI - Herpes simplex virus type 1 ICP27 is an essential regulatory protein. AB - The five immediate-early genes of herpes simplex virus are expressed during the initial stages of the infectious cycle, and certain immediate-early proteins have been shown to play a regulatory role in subsequent viral gene expression. Until recently, the functional properties of only one immediate-early protein, ICP4, had been examined in any detail, primarily because mutants had been isolated only in the gene for ICP4. We report herein the genetic and phenotypic characterization of four temperature-sensitive mutants of herpes simplex virus type 1 (tsY46, tsE5, tsE6, and tsLG4) that have begun to elucidate the function(s) of a second immediate-early protein, ICP27. The four mutants complemented each other inefficiently or not at all, indicating that they are defective in the same function. Marker rescue tests placed the mutations in tsY46 and tsE5 in sequences that encode the transcript for ICP27; the mutations in tsE6 and tsLG4 lie in or near these sequences. The ability of wild-type ICP27 expressed from a cloned gene to complement tsY46 and tsLG4 constitutes additional evidence that these mutants are defective in an ICP27-associated function. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of mutant-infected cell polypeptides showed that certain immediate-early (alpha) polypeptides were overproduced, whereas significant levels of early (beta) and drastically reduced levels of several late (gamma) proteins were synthesized at the nonpermissive temperature. Interestingly, the mutants were observed to form a spectrum with regard to their relative abilities to induce the expression of a number of polypeptides, especially those of the delayed-early (beta gamma) class. Consistent with their ability to induce expression of early polypeptides, all of the mutants induced the synthesis of substantial levels of viral DNA at the nonpermissive temperature. Taken together, the results of these studies demonstrate that ICP27 plays an essential regulatory function in virus replication, that this function is required after the onset of early gene expression and viral DNA synthesis, and that the inability of the mutants to induce the synthesis of late proteins is independent of viral DNA synthesis. PMID- 2991597 TI - Decreasing the number of 68-base-pair tandem repeats in the BK virus transcriptional control region reduces plaque size and enhances transforming capacity. AB - Wild-type BK virus, which has three 68-base-pair (bp) elements tandemly repeated in its transcriptional control region, forms clear, large plaques in human embryonic kidney cell cultures but rarely transforms hamster or rat cells. From this BK virus DNA, deletion mutants with fewer than three 68-bp elements were constructed and characterized. The mutant DNA with two 68-bp elements formed small, turbid plaques in human cell cultures and transformed rat 3Y1 cells inefficiently. The mutant DNA with one 68-bp element formed minute, turbid plaques in human cell cultures and transformed rat cells efficiently. PMID- 2991598 TI - Complete sequence of bluetongue virus L2 RNA that codes for the antigen recognized by neutralizing antibodies. AB - The complete sequence of the RNA which encodes the major outer-shell-neutralizing antigen (VP2) of bluetongue virus serotype 10 was determined from overlapping cDNA clones inserted into pBR322. The segment L2 RNA was 2,926 base pairs long (1.87 X 10(6) daltons) and had, in one strand, an open reading frame capable of coding for a protein that had a calculated size of 111,122 daltons (956 amino acids) and a +11.5 net charge. The coding strands of both the L2 gene and the group-specific L3 gene of bluetongue virus serotype 17 (M. Purdy, J. Petre, and P. Roy, J. Virol. 51:754-759, 1984) had common sequences of some six nucleotides at their 5' termini (namely, GUUAAA...) and eight nucleotides (namely, ...ACACUUAC) at their 3' termini. Both had short 5' noncoding regions with AUG codons at residues 20 to 22 (L2) and 18 to 20 (L3). The sequences flanking these AUG codons were similar (A/GCCAUGG). The 3' noncoding regions were longer (36 nucleotides for L2, 49 nucleotides for L3). The predicted amino acid sequence of the L2, compared with the similarly sized L3 gene product, was rich in cysteine residues and charged amino acids. PMID- 2991599 TI - trans-Activation of the human T-cell leukemia virus long terminal repeat correlates with expression of the x-lor protein. AB - Cell lines established directly from adult T-cell leukemia-lymphoma patients or immortalized by human T-cell leukemia virus type I (HTLV-I) in vitro that do not produce complete HTLV virions were characterized both for the content of viral proteins and for the presence of trans-acting factors activating gene expression under the control of the HTLV long terminal repeat. The expression of the 42 kilodalton HTLV x-lor product correlated with trans-activation of the long terminal repeat. The implications of this study for understanding the role of the HTLV x-lor product in the initiation and maintenance of T-lymphocyte transformation are discussed. PMID- 2991600 TI - Induction of hepatitis A virus-neutralizing antibody by a virus-specific synthetic peptide. AB - Comparative surface feature analyses of the VP1 sequences of hepatitis A virus (HAV) and poliovirus type 1 allowed an alignment of the two sequences and an identification of probable HAV neutralization antigenic sites. A synthetic peptide containing the HAV-specific amino acid sequence of one of these sites induced anti-HAV-neutralizing antibodies. It is concluded that a structural homology exists between the two viruses, despite minimal primary sequence conservation. PMID- 2991601 TI - Cell-related sequences in the DNA genome of human cytomegalovirus strain AD169. AB - Human cytomegalovirus (HCMV) cloned EcoRI fragments R and b hybridized strongly, under standard high-stringency conditions, to uninfected cellular DNA of human, murine, or sea urchin origin. Less hybridization was detected with fragments, A, C, E, WL(F), WN(H), I, M, O, P, Q, V, c, d, and e. Southern blot analysis of the HCMV-related human DNA localized the major sites of hybridization of HCMV EcoRI fragments R, b, and d to defined regions of the 28S rRNA gene. PMID- 2991602 TI - Latent herpes simplex virus type 1 DNA contains two copies of the virion DNA joint region. AB - Southern blot analysis of latent herpes simplex virus DNA detected in mouse brain and digested with a restriction enzyme revealed two copies of the virion DNA joint fragment. Thus, the absence of free ends noted previously in latent herpes simplex virus type 1 DNA is due to joining of the termini. PMID- 2991603 TI - Temporal replication of the Pullman strain of Aleutian disease virus in royal pastel mink. AB - Information was sought on the temporal replication of Aleutian disease virus in 27 royal pastel mink. Groups of three were examined 8 to 126 days after they were inoculated subcutaneously with 10(3) 50% lethal doses of the Pullman strain. Much individual variation was noted in the onset of infection, occurrence of viremia, and extent of virus replication in the tissues. Thus, virus was detected in lymph nodes regional to the site of inoculation in only some mink during the first 14 days after inoculation. During this period, virus was often present as well in the mesenteric lymph node and spleen. First detected on day 10, viremia was present in all mink examined on day 28 but occurred irregularly thereafter, even when virus was widespread in the tissues. Except in five mink succumbing to the disease, the tissue distribution of virus after day 28 tended to be more limited, and the titers were generally lower than they had been earlier. Even though present in the lymph nodes and spleen, virus was often absent from the kidney, liver, and intestine after day 28. Specific antibody was detected on day 28 and was present in all mink thereafter, ostensibly without any adverse effect on virus replication. In most mink, the infection was considered subclinical, for it was usually not accompanied by a rise in serum gamma globulin or by morphologic evidence of the disease. The virologic findings in this study have a bearing on the relationship of subclinical infections to both horizontal and vertical transmission of the virus. PMID- 2991604 TI - Binding of complement component C3b to glycoprotein C is modulated by sialic acid on herpes simplex virus type 1-infected cells. AB - Neuraminidase treatment of cells infected with herpes simplex virus type 1 (HSV 1) markedly enhanced the binding of complement component C3b to HSV 1 glycoprotein C (gC). When HSV-1 was grown in BHK RicR14 cells in which glycoproteins had reduced amounts of N-linked complex oligosaccharides, including sialic acid, the binding of C3b to gC was markedly enhanced. We used neuraminidase treatment to demonstrate that cloning the gC gene from the HSV-1 F strain into an HSV-1 mutant which fails to express gC converted the mutant virus from C3b receptor negative to receptor positive. These results further support a role for gC as a C3b receptor and indicate that sialic acid modifies receptor activity. PMID- 2991605 TI - Tissue selectivity of murine leukemia virus infection is determined by long terminal repeat sequences. AB - Here we show that the tissue specificity of murine retrovirus infections is determined by the long terminal repeat (LTR) of an otherwise isogenic set of viruses. The isogenic viruses used for this study contain the coding gag, pol, and env genes of the avirulent Akv virus. Recombinant viruses that contain the LTR of a virus that induces T-cell leukemia lymphoma preferentially infect T lymphocytes. Viruses that carry the LTR of a virus that induces erythroleukemia preferentially infect non-T lymphoblastoid cell lines in the marrow and spleen. The Akv virus itself displays no tissue preference for hematopoietic cells. These experiments suggest that retroviruses that carry appropriate enhancer-promoters can be used to infect selectively specific target cells in animals. PMID- 2991606 TI - Enhancement of the transforming capacity of BK virus by partial deletion of the 68-base-pair tandem repeats. AB - We constructed a set of deleted BK viral DNAs that lack part of the triplicated 68-base-pair (bp) sequences (the central unit has an 18-bp deletion) and assayed them for capacity to transform hamster kidney cells. Deletion of one of the two 68-bp units remarkably enhanced transforming capacity. PMID- 2991607 TI - Translational readthrough of an amber termination codon during synthesis of feline leukemia virus protease. AB - Feline leukemia virus contains a protease which apparently has the same specificity as murine leukemia virus protease. It cleaves in vitro the Pr65gag of Gazdar-mouse sarcoma virus into the constituent p15, p12, p30, and p10 proteins. We purified the protease and determined its NH2-terminal amino acid sequence (the first 15 residues). Alignment of this amino acid sequence with the nucleotide sequence (I. Laprevotte, A. Hampe, C. H. Sherr, and F. Galibert, J. Virol. 50:884 894, 1984) reveals that the protease is a viral-coded enzyme and is located at the 5' end of the pol gene. As previously found for murine leukemia virus (Y. Yoshinaka, I. Katoh, T. D. Copeland, and S. Oroszlan, Proc. Natl. Acad. Sci. U.S.A. 82:1618-1622, 1985), feline leukemia virus protease is synthesized through in-frame suppression of the gag amber termination codon by insertion of a glutamine in the fifth position, and the first four amino acids are derived from the gag gene. PMID- 2991609 TI - Unusual features of the leader sequence of Rous sarcoma virus packaging mutant TK15. AB - TK15, a mutant derived from a temperature-sensitive mutant of Rous sarcoma virus (tsNY68), has extremely low infectivity although it has intact viral genes. Biological and biochemical analyses of the virus and virus-induced transformants showed that the mutant has a defect in the packaging of its own genomic RNA possibly owing to a deletion near the 5' end (S. Kawai and T. Koyama, J. Virol. 51:147-153, 1984; T. Koyama, F. Harada, and S. Kawai, J. Virol. 51:154-162, 1984). Nucleotide sequence analysis of the provirus DNA of the mutant revealed that the deletion extends from the 3' end of the primer binding site to 22 bases upstream of the gag initiation codon and also suggested that possible binding between an extra region of the primer molecule and the viral genome resulting from the deletion causes another defect in the replication of the TK15 genome. It was suggested that the deletion of 237 bases in TK15 was generated during reverse transcription of the genome by the skipping of a sequence between identical 13 base sequences present in the primer binding site and 35 to 22 bases upstream of the gag initiation codon of the parental virus. PMID- 2991608 TI - Use of site-specific antipeptide antibodies to perturb the serine kinase catalytic activity of p37mos. AB - The mos oncogene of Moloney murine sarcoma virus encodes a protein of approximately 37,000 daltons (designated p37mos). We have detected a serine protein kinase activity which is closely associated with p37mos in immune complexes obtained with antibodies [anti-mos(37-55) serum] that were generated with a peptide containing amino acids 37 through 55 of the v-mos protein (S. A. Maxwell and R. B. Arlinghaus, Virology 143:321-333, 1985). Immune complexes that were derived with antibodies generated against peptides representing the C terminal 8 or 12 amino acids of v-mos (anti-C2 and anti-C3 serum, respectively) exhibited very little kinase activity capable of phosphorylating p37mos. Treatment of anti-mos(37-55) complexes containing active v-mos kinase with anti C3 or anti-C2 serum resulted in a dramatic reduction of the in vitro phosphorylation of p37mos. Antiserum blocked with the appropriate C-terminal peptide had no inhibitory effect on the phosphorylation of p37mos in anti-mos(37 55) complexes which indicated that the inhibition of v-mos kinase activity was a specific effect of these antibodies. The specific inhibition of the in vitro phosphorylation of p37mos by antibodies directed against the C terminus of the v mos protein provides strong evidence that the v-mos gene encodes a serine protein kinase. In addition, the extreme C terminus of p37mos may be critical for an active v-mos kinase. PMID- 2991610 TI - Genetic determinants of resistance to ectromelia (mousepox) virus-induced mortality. AB - Resistance to ectromelia (mousepox) virus-induced mortality was examined in crosses between susceptible DBA/2J, A/J, and BALB/cByJ mice and resistant C57BL/6J and AKR/J mice. Depending on the cross, resistance to mousepox virus was shown to be determined by one or more independently assorting autosomal loci with dominant alleles for resistance in AKR/J and C57BL/6J mice and recessive alleles in A/J, BALB/cByJ, and DBA/2J mice. A sexual dimorphism in resistance to disease was also observed. PMID- 2991612 TI - Obstructive uropathy, renal dysplasia and nodular renal blastema: is there a relationship to Wilms tumor? AB - We report 3 cases of renal dysplasia related to congenital obstructive uropathy and associated with nodular renal blastema. The literature is reviewed in regard to this relationship, as well as the relationship of nodular renal blastema with Wilms tumor and Wilms tumor with congenital genitourinary anomalies. The possible implication of nodular renal blastema associated with dysplasia and obstructive uropathy is emphasized with respect to the risk of neoplastic change. PMID- 2991611 TI - The use of collagenase in the treatment of Peyronie's disease. AB - Purified clostridial collagenase was administered intralesionally in 31 men with Peyronie's disease. Within 4 weeks of treatment 20 patients showed an objective improvement. Pain was eliminated in 13 of 14 patients with this complaint at presentation within the same 4-week period. The ability to have intercourse was restored in 3 of 4 patients with this problem. Except for a small corporeal rupture at the site of injection in 1 patient, no significant untoward effects were noted. During the mean 9.8-month followup 1 recurrence of bending was noted. PMID- 2991613 TI - Adult nephroblastoma with glomerular intracapillary deposits of IgM in the contralateral kidney. AB - A 43-year-old man who presented with hematuria had a nephroblastoma in the right kidney. After nephrectomy the patient was treated with irradiation and chemotherapy. Despite the development of pulmonary metastases he remained reasonably well for several years. He died of metastatic disease 8 years after presentation. Histological study of the contralateral kidney at autopsy revealed intracapillary glomerular deposits of IgM. PMID- 2991615 TI - Changing concepts in the management of clinical stage I nonseminomatous testicular cancer: significance of prognostic factors. PMID- 2991614 TI - A pharmacological study of alpha adrenergic receptor subtypes in smooth muscle of human urinary bladder base and prostatic urethra. AB - Postsynaptic alpha adrenergic receptor subtypes mediating contraction of human urinary bladder base and prostatic urethra were investigated in vitro. Alpha adrenergic receptor agonists, noradrenaline and phenylephrine, induced contraction dose-dependently in these tissues. Alpha adrenergic receptor antagonists, prazosin and yohimbine, competitively antagonized the contraction induced by these agonists in the bladder base as well as prostatic urethra. The mean pA2 values for the antagonists in the bladder base and prostatic urethra were as follows: 8.89 and 8.96 for prazosin and 6.30 and 6.45 for yohimbine, respectively. Comparison of pA2 values of prazosin and yohimbine in both the tissues with those values found in the literature for these compounds in a variety of tissues containing alpha-1 or alpha-2 adrenergic receptor indicates that the postsynaptic alpha adrenergic receptor subtype in the human urinary bladder base and prostatic urethra is the alpha-1 type, not the alpha-2 type. PMID- 2991616 TI - Status report on the acquired immunodeficiency syndrome. Human T-cell lymphotropic virus type III testing. Council on Scientific Affairs. PMID- 2991617 TI - Epidemiologists also studying suicidal behavior. PMID- 2991618 TI - Unexplained lymphadenopathy in homosexual men. A longitudinal study. AB - In January 1982, the Centers for Disease Control began a prospective study of unexplained generalized lymphadenopathy syndrome in 78 homosexual or bisexual men. As of Aug 1, 1984, the median durations of lymphadenopathy and enrollment were 29.5 months and 20 months, respectively. Antibodies to human T-cell lymphotropic virus type III/lymphadenopathy-associated virus were present in 75 (96%) of the study patients. Five (6%) of the patients were diagnosed as having the acquired immunodeficiency syndrome five to 25 months after the onset of lymphadenopathy. At the time of their initial visit, these five patients had lower T-helper cell counts, hematocrits, lymphocyte counts, and pokeweed mitogen stimulation ratios and were more likely to report severe symptoms than patients who did not develop the acquired immunodeficiency syndrome. During 318 follow-up visits, except for the development of the acquired immunodeficiency syndrome in five patients, there has been little change in the initial laboratory and clinical measures of this cohort. Within the follow-up period of this study, the features of lymphadenopathy syndrome have been unremitting for the majority of patients. PMID- 2991620 TI - Leads from the MMWR. Isolation of HTLV-III/LAV from samples of serum proteins given to cancer patients--Bahamas. PMID- 2991619 TI - Absence of antibodies to HTLV-III in health workers after hepatitis B vaccination. AB - A proportion of the plasma for the triply inactivated, plasma-derived hepatitis B vaccine produced in the United States is obtained from homosexual men. Because homosexual men are a high-risk group for the acquired immunodeficiency syndrome (AIDS), concern has emerged that the vaccine could harbor the AIDS agent. To evaluate this risk, we tested 15-month postvaccination serum samples for antibodies to human T-cell lymphotropic virus type III in 100 health care workers who had received inactivated hepatitis B vaccine lots made from plasma collected between 1977 and 1979 and 100 who had received placebo injections. None of the 200 health workers had serological evidence of human T-cell lymphotropic virus type III infection. These serological findings lend additional support to earlier epidemiologic and immunologic observations suggesting that hepatitis B vaccine does not transmit infection with an AIDS virus. PMID- 2991621 TI - [Adrenocortical function in patients with valve surgery; evaluation of ACTH stimulation test]. PMID- 2991622 TI - [An anesthetic experience with a patient with Eaton-Lambert syndrome]. PMID- 2991624 TI - [Alteration of blood coagulation and fibrinolysis after transcatheter arterial embolization of hepatocellular carcinomas]. AB - Alteration in blood coagulation and fibrinolysis were observed serially after TAE 37 times by measuring PT, APTT, FDP, fibrinogen, AT III, plasminogen, and alpha 2 macroglobulin. Immediately after TAE, PT and APTT were prolonged, Fbg AT III, alpha 2-M, and platelets decreased, and FDP increased. These changes within 24 hours suggested that coagulation activities were accelerated, and fibrinolysis followed. Within four weeks after TAE, PT and APTT became most prolonged one to three days after, while AT III, Plg, and alpha 2-M became lowest on the third to seventh days. It is suggested that these decrease were mainly due to suppressed production resulting from damage of the residual liver. PMID- 2991623 TI - [Postoperative prophylactic irradiation of lung cancer]. AB - Eighty-three patients with resected lung cancer were studied to determine whether postoperative prophylactic irradiation improves survival. All patients had mediastinal lymph node metastases and underwent relative curative surgery. Thirty patients received postoperative irradiation, and another 52 were followed without further treatment. The 5-year survival rate was 19.2% in the surgery alone group and 20% in the postoperative irradiation group. No increase in survival time was noticed in the postoperative irradiation group. The 5-year survival rate was not influenced by histologic types, presence or absence of subcarinal lymph node metastases and right or left thoracotomy in either group. PMID- 2991625 TI - [A case of adenoid cystic carcinoma in the paranasal sinuses with 8-year survival after pulmonary metastasis-- clinicopathological reference to 6 non-squamous cell carcinoma cases in the paranasal sinuses]. AB - Most cases of paranasal sinuses cancer are histologically squamous cell carcinoma. However, adenocarcinoma is rarely found even among cases of submucosal mucous gland origin. As adenocarcinoma has tendencies to local recurrence and distant metastasis, the five-year survival rate of this type of tumor has been reported to be worse than that of squamous cell carcinoma. On the other hand, some investigators insist that the survival rate at 10 or 20 years should be discussed because of the slow-growing nature of this tumor. We report herein clinicopathological studies on six cases of adenocarcinoma of the paranasal sinuses, including one long survivor, a 63-year-old man who lived 11 years and 3 months after the first visit and 8 years after pulmonary metastasis. PMID- 2991626 TI - [Autologous bone marrow transplantation in a patient with lymphoma type adult T cell leukemia]. PMID- 2991627 TI - Estimation of rate of incidence of ATL among ATLV (HTLV-I) carriers in Kyushu, Japan. AB - From the mortality statistics on malignant lymphomas in Japan and nationwide studies on T- and B-cell malignancies, the number of deaths from adult T-cell leukemia/lymphoma (ATL) in adults older than 20 years in the Kyushu district was estimated at 339 or more per year in 1978-82. The incidence for ATL per 100,000 was estimated at 3.5 for adults and 5.7 for persons over 40 years of age in the Kyushu district. From the information from a nationwide study on the distribution of ATL virus (ATLV) carriers among volunteer blood donors, the number of ATLV carriers in adults older than 20 years in Kyushu was calculated at 561,000 (310,000-793,000 at 95% confidence limit) in recent years. The incidence of ATL per 1,000 ATLV carriers in Kyushu was calculated at 0.60 (0.43-1.09) for adults, and 0.75 (0.55-1.18) for those over 40 years of age which was very close to the independently calculated figure of 0.84 (0.75-0.95) in the Goto Islands (the most typical ATL-endemic area). From these results, it was suggested that one case of ATL developed per year among 1,700 (900-2,300) ATLV-carrier adults and among 1,300 (800-1,800) ATLV carriers older than 40 years in Kyushu in recent years. PMID- 2991628 TI - Factors affecting the rate of positivity of sputum cytology in lung cancer. AB - We studied the role of sputum cytology and factors influencing the rate of positivity of sputum cytology in 103 patients with lung cancer. Sputum cytology was positive in 71.8% of these patients. Of 647 specimens submitted for sputum cytology, 610 were evaluable and the positivity rate was 43.6%. Forty-eight patients had at least six sputum cytology examinations. The rate of positivity for each of these examinations was between 31.3% and 50.0%, and the cumulative rate of positivity in these 48 patients was 77.1% at the sixth examination. Factors influencing the positivity rate were the histology of the tumor, tumor size, the site of the tumor, admixture with blood and the degree of surface necrosis of the tumor. The histology suspected from the sputum cytology coincided well with that determined at the operation or necropsy in adenocarcinoma, squamous cell carcinoma and small cell carcinoma, but not in large cell carcinoma. PMID- 2991629 TI - [Significance of CEA in sputum in examination of lung cancer]. PMID- 2991630 TI - [Role of systemic chemotherapy in smalL cell lung cancer as a part of multidisciplinary treatment]. PMID- 2991631 TI - [Treatment of small cell carcinoma of the lung]. PMID- 2991632 TI - [Characterization of eccrine carcinoma established in tissue culture from a patient with epidermodysplasia verruciformis]. PMID- 2991633 TI - [S-100 protein-positive cells in sweat apparatus tumors]. PMID- 2991634 TI - [Clinical evaluation of the glucagon test in fatty liver]. PMID- 2991636 TI - Effects of angiotensin converting enzyme inhibitor and angiotensin II analogue on hypertension. Study of hypertension with two different blockers of the renin angiotensin-aldosterone system. PMID- 2991635 TI - Comparison of the acute effects of three structurally different converting enzyme inhibitors (captopril, SA-446 and MK-421) on blood pressure, the renin angiotensin system and the kallikrein-kinin system. PMID- 2991637 TI - [The usefulness of scrotal scintigraphy in acute intra-scrotal lesions with special reference to differential diagnosis of spermatic cord torsion from epididymitis]. PMID- 2991638 TI - Bronchopulmonary sequestration associated with aspergilloma, with special reference to the finding of radionuclide venoangiography. AB - A case of intralobar bronchopulmonary sequestration associated with aspergilloma was reported. It was suggested by radionuclide venoangiography and aortic cineangiography. The diagnosis was confirmed by subsequent pathologic examination of the lobectomized lung tissue. The pulmonary perfusion scan with 99mTc-MAA showed a perfusion defect in the right lower lung field. Perfusion defect could also be traced in the same anatomical location in the pulmonary phase of radionuclide venoangiography with 99mTc-pertechnetate. However, the area of perfusion defect was visualized as hot area, when the injected radionuclide was pumped into the aorta after its first pass through the lung. The radionuclide venoangiography can efficiently detect abnormal systemic blood supply of the lung tissue. It can be an useful method for suggestive diagnosis of bronchopulmonary sequestration. PMID- 2991639 TI - Feeding control through bioassay of body chemistry. PMID- 2991641 TI - Effects of clonidine, alpha-methyldopa and hydralazine on met-enkephalinergic neurons in cerebral nuclei of spontaneously hypertensive rats. AB - Immunocytofluorescent and microautoradiographic methods were applied to measure met-enkephalin-like immunoreactivity (MELI) and ME receptor binding (MERB) levels in cerebral nuclei of serial brain slices from SHR which received clonidine, alpha-methyldopa and hydralazine at equivalent hypotensive doses. All three drugs increased both MELI and MERB levels in the caudal part of the n. tractus solitarii and its functionally related dorsomedullary nuclei and decreased them in the n. accumbens septi, in accordance with the correspondent change in glucose utilization rates in these nuclei as reported previously. Both CNS-active agents (not hydralazine) also increased MELI and MERB levels in the n. intercalatus and substantia grisea centralis, and they decreased them in the n. ventromedialis hypothalami. Differences in both CNS-active agents were minor. Vasodilative hydralazine alone decreased these levels both in the n. reticularis medialis and n. tegmenti ventralis, and it increased them slightly in the area lateralis hypothalami. The present studies indicate that ME neurons of these dorsomedullary and supramedullary nuclei may act as direct as well as homeostatic controls of blood pressure. PMID- 2991640 TI - Influence of polymeric effectors on binding of 3H-dihydroalprenolol to beta adrenergic receptor of rat brain. AB - The significance of anionic and cationic charges of glycocalyx, phospholipid or protein, etc. on the cell surface of the rat brain was examined for beta adrenoceptors using the radioligand binding assay method. Thus, this experiment was designed to assess the effects of polymeric effectors, DNA, heparin, polymyxin B, histone, gelatin, colominic acid and bovine serum albumin (BSA), on the affinity of beta-adrenoceptors. The rat brain was used and the beta adrenoceptor binding assay was carried out using 3H-dihydroalprenolol as a radioligand. Polymyxin B, DNA and heparin significantly caused a reduction in the maximum number of beta-adrenoceptors (Bmax), but only small changes were observed with histone, gelatin, BSA and colominic acid. Only DNA induced a decrease in the value of the dissociation constant (Kd) of beta-adrenoceptors. These results suggest that anionic or cationic charges in the environment of the receptor sites could have a crucial role in drug-receptor interaction. PMID- 2991642 TI - [Acquisition of beta-adrenergic blockade by repeated induction of airway anaphylaxis: changes in beta-adrenergic receptor levels in lung homogenates and beta-adrenergic response in tracheal muscles in guinea pigs with experimental asthma]. PMID- 2991643 TI - [Surgical indications of and their limitations in pulmonary cancer with special reference to lymph node metastasis]. PMID- 2991644 TI - [CT diagnosis of mediastinal lymph node metastasis]. PMID- 2991645 TI - [Surgical correction of chest deformity in a 13-year-old girl with Poland syndrome]. PMID- 2991646 TI - Bimodal differentiation prospectives for promyelocytes. AB - In accordance with the long-range goal to demonstrate the direct derivation of blood monocytes from promyelocytes in human bone marrow, the promyelocytic cell line HL-60 in the present study was differently stimulated with various inducers for the purpose of documenting its capability to evolve into granulocytes or monocytes-macrophages. Each differentiation line was monitored with the use of marker enzymes, antigens, and cell-specific monoclonal antibodies. In addition, studies were done on normal human bone marrow and leukemias. The results provided strong evidence for the close cytogeneic relationship of granulocytes and monocytes and for a common progenitor at the maturation stage of promyelocytes myelocytes for both cell lineages. PMID- 2991648 TI - [Clinical importance of 99mTc-pyrophosphate scintigraphy in the bicycle ergometry test]. AB - The clinical value of myocardial scintigraphy with 99mTc-pyrophosphate in conjunction with bicycle ergometry was assessed through the correlation of the latter's results with the clinical course of coronary heart disease over 1 year in 21 patients with first myocardial infarction during its early healing stage and 17 anginal patients without a history of myocardial infarction. The disease ran a stable course at the time of the testing in all the patients; later on, 11 of those developed various complications associated with coronary heart disease, or died. Three scintigraphic recordings were made: prior to, at the peak of, and after the exercise. Thirteen scintigraphic patterns were identified. They are interpreted on the basis of presumed variation in the degree of myocardial damage and blood supply to affected areas under the effect of exercise; a relation is also noted with predicted risk of complications. The method has proved reliable in identifying patients at low risk for complications. Providing indirect evidence of the quality of blood supply to damaged myocardial areas during rationed exercise, it furnishes additional criteria for the assessment of the patients' working capacity. PMID- 2991647 TI - Studies of methotrexate-monoclonal antibody conjugates for immunotherapy. AB - Methotrexate (MTX) was covalently bound to two different murine monoclonal antibodies, one reactive with human colon carcinoma and the other reactive with the transferrin receptor. The drug-antibody complexes were examined for their in vitro and in vivo potency against tumors. The conditions of coupling were closely monitored, with particular attention being paid to the preservation of both drug and antibody activity. After activation of MTX with N-hydroxysuccinimide, MTX was bound to the antibodies under conditions leading to maximum protein and antibody recovery. Although the coupling conditions for both antibodies were different, up to 13 molecules of MTX-antibody molecule could be attached with retention of antibody activity. Such conjugates were active in vitro and could inhibit the growth [( 3H]deoxyuridine uptake) of cells in culture. The conjugates were highly specific, having no effect on tumors lacking the antigen; however, MTX complexed to antibody was less potent than the free drug. In vivo, the MTX-antibody impaired the growth of established tumors. Thus MTX-antibody complexes can be successfully produced and can be used for the immunotherapy of tumors. PMID- 2991649 TI - [Nuclear medicine diagnosis of hemorrhaging Meckel's diverticulum using 99mTC pertechnetate--demonstrated by 2 cases]. PMID- 2991650 TI - [Treatment procedure in phyllodes tumors of the breast]. PMID- 2991651 TI - [Exogastric form of glomus tumor of the stomach]. PMID- 2991652 TI - [Metastasis of small-cell lung cancer to the appendix]. PMID- 2991653 TI - A teratoma group tumor of testis with mature teratoma in metastatic foci--a case report. PMID- 2991654 TI - Arginine deficiency, hyperammonemia and Reye's syndrome in ferrets. AB - Young male ferrets developed hyperammonemia and encephalopathy shortly after eating a diet lacking in arginine. The dietary supplementation of arginine or intraperitoneal injection of ornithine prevented hyperammonemia and shortened the duration of encephalopathy. Therefore, young ferrets were assumed to be unable to meet their ornithine needs from sources other than arginine. Adult ferrets did not develop hyperammonemia and encephalopathy after eating arginine-free diet. Because young ferrets are also susceptible to human influenza infections, they were further tested as animal model of Reye's syndrome. Reye's syndrome is a serious childhood disorder that develops following influenza infections and is characterized in part by an encephalopathy, hyperammonemia and elevated serum transaminases. In young ferrets, concurrent administration of aspirin with human influenza inoculation and an arginine-free diet produced symptoms similar to those seen in humans with Reye's syndrome. The ferret model appears to be useful for studying the roles of various etiologic agents and their interactions in producing Reye's syndrome-like disorders. The ammonia metabolism in ferrets is reviewed and the ferret model for Reye's syndrome and its applications for the better understanding of this disorder in humans are discussed. PMID- 2991655 TI - Ultrastructural comparison of the retroviruses associated with human and simian acquired immunodeficiency syndromes. AB - Three isolates of human acquired immunodeficiency syndrome (AIDS) virus (lymphadenopathy-associated virus, human T cell lymphotropic virus strain III, acquired immunodeficiency syndrome-associated retrovirus) were compared with simian AIDS virus by thin section transmission electron microscopy. Simian AIDS virus was found to form intracytoplasmic A particles and to bud by both ring forms and crescent shapes. The extracellular mature simian AIDS virus exhibited a cylindrical nucleoid within a tubular or conical core shell. The simian AIDS virus was morphologically indistinguishable from Mason-Pfizer monkey virus, the prototype D type retrovirus, although the two were readily distinguished by radioimmunoassay and restriction endonuclease mapping. All three AIDS virus isolates exhibited identical ultrastructure and maturation sequences. The AIDS viruses did not form intracytoplasmic A particles and budded only with crescent shaped nucleoids. Mature extracellular AIDS viruses contained an eccentric spherical nucleoid within a conical or tubular core shell. AIDS virus was found to show many morphologic features of the retroviral subfamily Lentivirinae. The AIDS virus isolates appear to form a new group of retroviruses. PMID- 2991656 TI - Terminally differentiated derivatives of pulmonary small cell carcinomas may contain neurofilaments. PMID- 2991658 TI - Metabolism of estradiol by true and pseudoperoxidases. AB - The activation of 14C-labeled estradiol by "true" and "pseudo" peroxidases to form conjugates and other products was compared in four model systems using H2O2, glutathione, Mn2+ or irradiated riboflavin. Albumin was used as acceptor except in the glutathione system. The binding of estradiol to glutathione in the presence of the true peroxidases, lacto- or uterine peroxidase (no H2O2 added), was also examined and the conditions shown to differ from those required with the pseudoperoxidases, microperoxidase or trypsin-digested cytochrome c. The conjugates were purified by chromatography after elution from Amberlite XAD-2 and the relative amounts of these products assessed by autoradiography. The ratio of steroid to glutathione in the main water-soluble metabolite formed with lactoperoxidase was found to be approx 1:1 in a double label experiment with [14C]estradiol and [3H]glutathione. It was also shown, using estradiol labeled with 3H in different positions of the steroid molecule, that lactoperoxidase acts non-specifically in catalyzing the formation of glutathionyl conjugates as indicated by the release of 3H2O. The possible role of peroxidase and glutathione in the metabolism of estrogens and in the formation of artifactual products is discussed. PMID- 2991657 TI - Rapid recovery of nuclear estrogen receptor and oxytocin receptor in the ovine uterus following progesterone withdrawal. AB - We previously showed that progesterone rapidly down regulates nuclear estrogen receptor (Re) in the estrogen-primed rodent uterus. We have now extended these studies to test the response of the Re system in sheep uterus to progesterone withdrawal. Since the estrogen-Re complex is believed to regulate hormone dependent gene expression, it was of interest to determine whether withdrawal of progesterone under constant estrogen stimulation would lead to the recovery of nuclear Re levels and estrogen action, i.e. oxytocin receptor (ROT) synthesis. Ovariectomized ewes were primed with estradiol-17 beta and serum steroid levels were maintained by constant infusion of estradiol (0.5 microgram/h) and progesterone (500 micrograms/h) for 5 days. The animals were anesthetized with fluothane/O2, and uterine samples were excised 1 h before and 3, 6 and 12 h after progesterone withdrawal. Estradiol infusion was continued during the experiment in order to maintain estrogen levels at a steady state (14 pg/ml plasma). Re, ROT and progesterone receptor (Rp) were measured in endometrium and myometrium using standard 3H-hormone binding assays. Following progesterone withdrawal, the nuclear Re concentration increased in both uterine compartments, and the nuclear Re level was correlated significantly with the ROT concentration in the membrane fraction of both uterine tissues (endometrium, r = 0.79; myometrium, r = 0.86). Although cytosol Re rose between 6 and 12 h in the endometrium, cytosol Re levels remained unchanged in myometrium. Cytosol Rp appeared to increase in endometrium but not in myometrium. Uterine tissue sampled from a control animal before stopping the progesterone infusion revealed that the observed changes in receptor concentration following progesterone withdrawal were not due to regional differences in receptor levels. These results demonstrate that the recovery of nuclear Re in the ovine endometrium and myometrium following progesterone withdrawal represents a selective effect on Re retention in the nucleus rather than on cytosol Re availability or Re activation which was controlled by constant estrogen infusion. Thus, these results are consistent with the hypothesis that progesterone induces an Re regulatory factor which acts to down regulate nuclear Re, and that the activity of this factor diminishes rapidly after progesterone withdrawal. PMID- 2991659 TI - Are polyphosphorylated phospholipids involved in the hormonal control of cholesterol side-chain cleavage activity in tumour Leydig cells? AB - The possible role of LH or dcAMP induced changes in polyphosphorylated phospholipid metabolism in the regulation of cholesterol side-chain cleavage activity has been studied in tumour Leydig cells. Mitochondria isolated from LH stimulated Leydig cells were 400% more active in pregnenolone production than mitochondria from control cells. Steroid production in isolated mitochondria from control cells could be stimulated only 25% by cytosol fractions from stimulated cells and 100 microM phosphatidyl inositol-4'-phosphate (PtdIns4P). Other polyphosphorylated phospholipids were either inactive or showed aspecific effects. During a preincubation period tumour cells were labelled with [32P]phosphate and steady-state labelling was obtained for the pholyphosphorylated phospholipids after 40-60 min. [32P]Phosphate incorporation in Ptd Ins4P, phosphatidyl inositol (PtdIns), phosphatidyl choline (PtChl), phosphatidyl ethanolamine (PtdEtn) and cardiolipin (CL) was not affected by treatment of the Leydig cells with LH which stimulated (6-fold), or with cycloheximide which suppressed (4-fold) steroid production. A 25% increase of phosphate incorporation by LH was observed only in phosphatidyl inositol-4',5' biphosphate (PtdIns4,5P2). 32P Incorporation in PtdIns4,5P2, PtdIns,PtdEtn and CL was stimulated by quinacrine 50 microM. Under these conditions the LH-stimulated pregnenolone production but not the 25-hydroxycholesterol dependent pregnenolone production, was completely inhibited. The results obtained with isolated mitochondria and intact cells indicate that increased levels of polyphosphorylated phospholipids are not consistently correlated with increased mitochondrial pregnenolone production. This argues against an important role of polyphosphorylated phospholipids in the hormonal regulation of cholesterol side chain cleavage activity in tumour Leydig cells. PMID- 2991660 TI - 3 Beta-hydroxysteroid dehydrogenase-isomerase activity in bovine adrenocortical cells in culture: lack of response to ACTH treatment. AB - Primary cultures of bovine adrenocortical cells (BAC) were used to determine whether the adrenal microsomal 3 beta-hydroxysteroid dehydrogenase-isomerase complex (3 beta-HSD), like the 17 alpha-hydroxylase (17-OHase), responded to ACTH treatment with an increase in activity. Both enzymes influence the steroidogenic path leading to 17 alpha-hydroxyprogesterone formation and thus could affect adrenal androgen biosynthesis. 3 beta-HSD Activity in postmitochondrial supernatant fluid, homogenates or cell monolayers remained unchanged after cells had been maintained in 1 microM ACTH up to 48 h. Since ACTH exposure led to a marked increase in 17-OHase activity over the same time period, it is concluded that, under the conditions used, the 3 beta-HSD-isomerase complex in BAC is nonresponsive to tropic hormone treatment. PMID- 2991661 TI - Effects of hyperprolactinemia on activities of 17-hydroxylase, 17 beta-ol dehydrogenase and 5 alpha-reductase in neonatally grafted and host testes in mice. AB - Male and female (WB X C57BL/6)F1 hybrid mice were used. Two pituitaries from 60 80-day-old female mice were grafted under the capsule of the left kidney of 60-80 day-old male mice. One week after grafting, 2 testes from neonatal mice were grafted under the capsule of the right kidney of the grafted mice and 70-90-day old intact male mice. The grafted and host testes, in groups of 10-26, were removed 15, 30, 40, 60 and 120 days after transplantation of the neonatal testes. Testicular homogenates were incubated with [3H]progesterone or [14C]4-androstene 3,17-dione, and enzyme activities per g tissue were estimated. Significantly elevated prolactin levels, slightly lower LH levels and normal testosterone levels were found in the mice with pituitary grafts, compared with those in the mice without pituitary grafts. Activities of 17-hydroxylase and 17 beta-ol dehydrogenase increased clearly with age in the grafted testes in the mice without pituitary grafts, though the increases were inhibited significantly by the pituitary grafts. However, the pituitary grafts had no significant effect on activities of 17-hydroxylase and 17 beta-ol-dehydrogenase in the host testes under similar gonadotrophic stimulation. 5 alpha-Reductase activities in the grafted and host testes were unaffected by the pituitary grafts. These results show that hyperprolactinemia may directly inhibit increases in activities of 17 hydroxylase and 17 beta-ol-dehydrogenase with testicular age in neonatally grafted testes in mice. PMID- 2991662 TI - Appearance of the rat testicular receptor for calcitriol (1,25-dihydroxyvitamin D3) during development. AB - In the present study we have examined the developmental changes in the concentration of receptors for calcitriol in high-salt cytosol from the rat testis. Receptors for calcitriol were undetectable (less than 0.4 fmol/mg protein) until day 24, after which there was a rapid increase to reach adult levels (6-8 fmol/mg protein) between day 50-60. The lack of receptors in high salt cytosol from the immature rat testis is not due to degrading enzymes, since cytosols prepared from the combination of equal volumes of testis homogenates from immature and adult rats had binding levels exactly half of that found in "adult controls". Furthermore, the increase in specific binding of [3H]calcitriol during development is due to an increase in the number of receptor sites, and is not due to a change in the apparent affinity of the receptors (Kd approximately equal to 1 X 10(-11) M at 0 degrees C). These results may explain why we previously were unable to demonstrate calcitriol receptors in cultured Sertoli cells and peritubular cells isolated from 19-day old rats. Furthermore, they indicate that calcitriol may be of minor importance for testicular function in the immature rat. The role of calcitriol in the pubertal and adult testis remains to be established. PMID- 2991663 TI - The transmethylations inhibitor 3-deazaadenosine, inhibits in vitro testosterone production by rat testis interstitial cells stimulated with HCG. AB - 3-Deazaadenosine (3-DZA), an inhibitor of somatic cell transmethylations, inhibited in vitro HCG-stimulated testosterone synthesis by rat testis interstitial cells. A maximal inhibition of 50% was observed with 100 microM 3 DZA; in addition homocysteine-thiolactone (Hcy) enhanced the inhibitory effect of 3-DZA. On the other hand, when cells were stimulated with dibutyryl cyclic AMP (Bt)2-cAMP, 3-DZA did not exert any effect on the stimulation. The presence of 3 DZA in the incubation medium neither modified HCG Kd values nor the number of its binding sites to Leydig cells. These results demonstrate that inhibitors of transmethylation reactions interfere with hormone-stimulated testosterone synthesis, suggesting that those reactions (presumably phospholipid methylation) at the plasma membrane level are involved in hormone-stimulated testosterone synthesis by rat Leydig cells. PMID- 2991664 TI - Synthesis and biologic activity of 3 beta-thiovitamin D3. AB - We synthesized 3 beta-thiovitamin D3 from 7-dehydrocholesterol and tested its biological activity and protein binding properties. The thiovitamin was found to be a weak vitamin D agonist at high doses in vivo. It was poorly bound by both vitamin D-binding protein as well as by the intestinal cytosol receptor for 1,25 dihydroxyvitamin D. It did not increase the synthesis of calcium binding protein in the chick embryonic duodenum and did not block the activity of 1,25 dihydroxyvitamin D3 in this system. We conclude that 3 beta-thiovitamin D3 is a weak vitamin D agonist in vivo with no agonist activity or antagonist activity to 1,25-dihydroxyvitamin D3 in the chick embryonic duodenum. PMID- 2991665 TI - The quantification of relative efficacy of agonists. AB - While much pharmacological effort has been expended in the measurement, quantification, and comparison of agonist affinity for the classification of drugs and drug receptors, inordinately less emphasis has been placed on the quantification of the other property of agonists, namely intrinsic efficacy. This is unfortunate as the existing studies of the relative intrinsic efficacy of agonists show this to be a most useful scale for the classification of agonists and the prediction of tissue responses. This paper will review some of the theories that describe efficacy on a molecular level, the methods of measuring relative efficacy, and the factors in these procedures which can lead to artifacts and misleading information for the classification of drug receptors. Lastly, the value of the quantification of efficacy will be discussed in terms of the design of agonists for therapeutic advantage. PMID- 2991666 TI - A model for transposon-based eucaryote regulatory evolution. AB - This paper presents a compact model of the role of transposable elements in eucaryote evolution which, although forward looking, is consistent with both experimental results and theories of gene regulation. The model postulates that a principal factor in the emergence of the eucaryotes was the development of a symbiotic relationship between reverse transcribing transposable elements and RNA based gene regulation, which we will call structural symbiosis. Thus, although transposable elements follow their own evolutionary protocol, structural homologies between "cellular" and "viral" genomes result in selective mutagenesis, a situation where transposon mutations are permitted because they can result in phenotypic mutations of the regulatory process with reduced probability of deleterious mutation of structural genes. The incorporation of this scheme into the life cycle of higher organisms results in two forms of integral evolution. Exogenous, in which differing species in an ecosystem share genetic information through viral transfer, and endogenous in which somatically induced regulatory mutations can be mapped back into the germ line. PMID- 2991667 TI - Prognostic evaluation of tracheobronchial reconstruction for bronchogenic carcinoma. AB - Fifty-two patients have undergone tracheobronchial reconstruction for bronchogenic carcinomas over a 20 year period and have been evaluated from the view point of prognosis. Five-year survival rates of the patients undergoing reconstructive operations were as follows: 35% for the total group, 50% for those with squamous cell carcinoma, and 64% for those with Stage I and II disease. No patients with adenocarcinoma or Stage III disease have survived more than 5 years. However, the number of patients with early adenocarcinoma was too small for us to conclude that the histologic type per se affected survival. Six of eight patients with sleeve lobectomy and pulmonary artery reconstruction died within 2 years, 7 months postoperatively. Five of seven patients died within 1 year after carinal reconstruction. However, two are alive at 4 months and 2 years, 9 months after left or right sleeve pneumonectomy. In summary, any types of lobectomy or pneumonectomy with reconstruction of the tracheobronchial tree can be conducted in patients with Stage I and II lung cancer. Sleeve lobectomy with pulmonary artery reconstruction can be an alternative to pneumonectomy when pneumonectomy is contraindicated because of low cardiopulmonary reserve. In patients undergoing reconstruction of the carina, prophylactic radiation therapy may be necessary during the postoperative course. PMID- 2991668 TI - Bronchoplastic procedures and pulmonary artery reconstruction in the treatment of bronchogenic cancer. AB - Nineteen patients with primary bronchogenic carcinoma underwent bronchoplastic procedures (six wedge and 13 sleeve resections) between 1970 and 1982. In six of them lobectomy was combined with sleeve resection and reconstruction of the pulmonary artery: In one a synthetic prosthesis was inserted. Twelve patients had squamous cell carcinoma, five adenocarcinoma, and two large cell carcinoma. No operative deaths were observed, and the 5 year survival rate is 28.1%. One patient had an early bronchial fistula and two patients had bronchial stenosis (one suture granulation and one local recurrence). No patient with resection of the pulmonary artery had vascular complications. Survival rates on the basis of nodal involvement indicate 50% survival at 5 years without nodal metastasis (11 cases) versus 9.7% with nodal involvement (eight cases) (p less than 0.05). Bronchoplastic procedures, even if accompanied by segmental resection of the pulmonary artery, can be performed safely with long-term results comparable to those following major pulmonary resections. PMID- 2991669 TI - Phorbol ester-induced loss of colchicine ultrasensitivity in chronic lymphocytic leukaemia lymphocytes. AB - On exposure to the phorbol ester 12-O-tetradecanoyl-13-acetate (TPA) the pathological (non-dividing) lymphocytes of B-cell chronic lymphocytic leukaemia (CLL) lose their characteristic ultrasensitivity to the cytocidal action of colchicine in vitro. They are no longer killed in 1 day by the drug at 10(-6)M concentration. The effect was the same whether the cells were incubated in the continuous presence of TPA, or subjected instead to pulse-treatment with it (for as little as 5 min.). Colchicine at one thousand times greater concentration was now needed to kill the cells. CLL lymphocytes already primed to undergo interphase death by pretreatment with colchicine could be prevented from doing so by early addition of TPA. A marked proportion of those CLL lymphocytes destined to undergo early spontaneous death in vitro in the absence of colchicine could be prevented from doing so by TPA. The loss of colchicine ultrasensitivity applied to cells which had not yet undergone TPA-induced morphological transformation to blast-like cells or differentiation to cells containing abundant cytoplasmic immunoglobulins (CIg). These transformed cells materialised in greatest incidence (70-80%) after 3 days of culture, an observation in agreement with others workers. PMID- 2991670 TI - Granulocyte functions during maturation of human promyelocytic leukemia cells. AB - Human promyelocytic leukemia cells (HL-60) mature into functional granulocytes in response to in vitro treatment with several classes of chemical agents. Compounds that increase intracellular adenosine 3':5' -cyclic monophosphate induce a modified program of maturation in which the cells demonstrate functional properties characteristic of mature phagocytic cells while remaining morphologically immature. We compared the developmental programs initiated by two well-studied inducers, retinoic acid and dimethyl sulfoxide, with the programs initiated by two inducers known to raise intracellular adenosine 3':5' -cyclic monophosphate; N6, O2-dibutyryl adenosine 3':5' -cyclic monophosphate and the combination of prostaglandin E2 and theophylline. In response to the increase in intracellular adenosine 3':5' -cyclic monophosphate, the cells ceased proliferation, expressed chemotactic receptors and demonstrated stimulated enzyme release within 24 h. Chemotaxis, adherence, NBT reduction and superoxide production appeared by 72 h, although the cells remained unchanged morphologically. A similar developmental program was induced by dimethylsulfoxide, but appearance of the markers was delayed by 48 h. Expression of these markers was delayed and incomplete in response to retinoic acid. PMID- 2991671 TI - Response of the tracheobronchial epithelium to hemoprotein tracers. PMID- 2991672 TI - Monoclonal antibody for rapid laboratory detection of cytomegalovirus infections: characterization and diagnostic application. AB - Monoclonal antibodies to early (2H2.4, molecular weight 72,000 daltons) and late (2F3.0, molecular weight 68,000 daltons) antigens of the AD-169 strain of cytomegalovirus (CMV) were prepared by fusing mouse spleen cells with NS-1 mouse myeloma cells. The 2H2.4 monoclonal antibody produced a dense immunofluorescence with prominent lobular staining within the nucleus of CMV-infected substrate cells, whereas the reaction of 2F3.0 was more diffuse and generally involved the entire nucleus of the cells. Both monoclonal antibodies had little or no neutralizing activity against CMV in plaque-reduction assays. No cross-reactions were observed between these monoclonal antibodies and other members of the herpesvirus group. The 2H2.4 monoclonal antibody to early CMV antigen was used in a shell vial assay with a low-speed centrifugation step for the rapid (within 16 hours after inoculation) diagnosis of CMV infections. Optimal conditions for the test included centrifugation of shell vials at 700 X g for 45 minutes at 36 degrees C. An inoculum volume of 0.2 ml provided a reasonable balance between the optimal sensitivity for detecting specific viral fluorescence and the easy discrimination of the specific immunofluorescence from the background debris. Because of the commercial availability of the monoclonal antibody and the simplicity of the procedures used in the shell vial assay and subsequent fluorescence techniques, this rapid assay can be done in any laboratory that is familiar with cell culture manipulations. PMID- 2991674 TI - Age-related reductions in rat atrial high affinity choline uptake, ACh synthesis, and ACh release. A brief note. AB - We have developed a rat atrial mince preparation that can take up choline, acetylate it, and then release acetylcholine in a depolarization-dependent manner. We demonstrate that aging appears to reduce the functional cholinergic activity in this tissue, which may be important for understanding how senescence alters the regulation of cardiac activity. PMID- 2991673 TI - Restriction enzyme analysis of mitochondrial DNA in aging human cells. AB - Human diploid fibroblasts show a limited lifespan in vitro. To investigate the integrity of mitochondrial DNA (mtDNA) in aging fibroblasts, whole cell DNA samples from the human cell line IMR-90 have been prepared at 36, 22, and 3 population doublings (PD) from the end of the lifespan (63 PD). These DNA samples were then digested separately with 19 different restriction endonucleases, and the resulting fragments were separated by agarose gel electrophoresis and transferred to nitrocellulose filters. Fragment sizes were revealed by hybridization to 32P-labelled mouse mtDNA and autoradiography, and were compared with computer maps of fragments generated from the known sequence of human mtDNA. These 19 enzymes recognize a total of 297 recognition sites comprising 1315 nucleotide base pairs (bp), approximately 8% of the human mtDNA (16 569 bp). Control experiments reveal that a minor component representing as little as 5% of the total mtDNA can be detected. No changes were seen in the restriction fragment pattern with fibroblast cell age. It is concluded that there are no large deletions, insertions, or rearrangements in human mtDNA, and no single base changes in the detectable regions. This suggests efficient maintenance of mtDNA molecules and/or elimination of damaged mtDNA during fibroblast cell lifespan. PMID- 2991675 TI - Binding of thyroid hormones by nuclei of target tissues during the chick embryo development. AB - In the present studies we have compared the ontogeny of the binding of thyroxine (T4) and triiodothyronine (T3) to isolated nuclei from various target tissues of chick embryo. We observed a marked difference between the patterns of Satchard plots, maximal binding capacities (MBC) and association constants (Ka) of T4 and those of T3. Scatchard plots revealed that T4 and T3 had different binding sites. In liver, brain and lung MBCs and Kas of T3 and T4 were rather similar at day 9, but during the following days (12-19) T3 MBCs and Kas showed small changes, whereas T4 MBC markedly increased (4-5-fold) and T4 Ka significantly declined. In liver, for instance, T3 MBC = 395 +/- 19 (day 9) and 489 +/- 66 fmol/mg protein (day 19); T4 MBC = 631 +/- 6.5 (day 9) and 2201 +/- 516 fmol/mg protein (day 19); T4 Ka = 1.92 +/- 0.01 (day 9) and 0.56 +/- 0.21 X 10(8) M-1 (day 19). These data indicate that, during chick embryogenesis, nuclei of target tissues contain multiple T4 binding proteins, but only a single T3 binding site. PMID- 2991676 TI - [Extrahepatic obstructive jaundice as the 1st manifestation of bronchogenic carcinoma]. PMID- 2991677 TI - [Tumoral hypophosphatemic osteomalacia]. PMID- 2991678 TI - Binding of a new opiate antagonist, nalmefene, to rat brain membranes. AB - Nalmefene (6-methylene-naltrexone) is a potent, orally active, opiate antagonist. IC50's were obtained for nalmefene, naloxone and naltrexone using radiolabelled prototype ligands for mu, kappa and delta receptors in homogenates of rat brain minus cerebellum. Nalmefene antagonized the bindings of [3H]-dihydromorphine, [3H]-ethylketocyclazocine and [3H]-D-ala-D-leu enkephalin with IC50's in the low nanomolar range. At the central mu receptor, nalmefene bound with an IC50 of 1.0 nM, equal to that of naltrexone and approximately four times lower than that of naloxone. At central kappa and delta sites the IC50's for nalmefene were somewhat lower than those of naltrexone and considerably lower than those of naloxone. All three antagonists had sodium indices less than 1.0. These results indicate that nalmefene is a universal opiate antagonist, has no agonist character at the central mu site and binds more effectively to central opiate receptors than either naloxone or naltrexone. PMID- 2991679 TI - Resection of upper aerodigestive tract tumors involving the middle cranial fossa. AB - Benign and malignant neoplasms of the upper aerodigestive tract that invade the middle cranial fossa are frequently considered unresectable due to the proximity of the cavernous sinus and internal artery, and to the inaccessibility of this region via conventional surgical approaches. We report our experience using a combined, lateral intracranial and infratemporal fossa procedure for the management of these tumors in four patients. Successful removal of neoplasm associated with minimal morbidity was accomplished in 3 patients, 2 with angiofibromas and 1 with an adenoid cystic carcinoma. The postoperative course of the fourth patient was complicated by meningitis which resulted in the patient's death five months following resection of an adenocarcinoma. PMID- 2991680 TI - Evaluation of NMR versus CT for parotid masses: a preliminary report. AB - Twelve patients with various parotid masses were prospectively studied by both nuclear magnetic resonance (NMR) imaging and intravenously enhanced computed tomography (CT). These imaging techniques were then correlated with the disease process and clinical relevance of both forms of imaging assessed. Our results suggest high resolution, spin-echo NMR offers several advantages over CT, and is likely to become the preferred method of evaluating a parotid mass. PMID- 2991682 TI - [Brain opiates and pain]. PMID- 2991681 TI - A clinical pathologic study of hearing loss in congenital cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is currently reported as the most common cause of congenital viral induced deafness. However, few systematic studies of the audiovestibular sequelae of this infection are present in the literature. A clinical pathologic study was conducted from 1976 to 1982 to evaluate this. Fifty two pairs of infant and children's temporal bone studied demonstrated no evidence of CMV endolabyrinthitis even in the single case with evidence of extensive congenital CMV infection. Over 2,000 umbilical cord sera were screened to detect asymptomatic CMV infection with an incidence of 0.38% (and slightly greater than 1% when extrapolated to correct for the sensitivity of the method of detection) in a central Pennsylvania study population. No sensorineural abnormalities were detected in five asymptomatic children and 30 control children. However, three out of six (50%) infants, symptomatic at birth and followed to a mean age of 5.5 years, showed significant and progressive sensorineural loss and vestibular deficits. PMID- 2991683 TI - Studies on the up regulation of alpha-adrenoceptors on rat hippocampal perikarya by chemical lesion of the median raphe nucleus. AB - Neurotoxin-induced lesion of the serotonergic raphe-hippocampal pathway produced about a 50% increase in the density of a nM affinity alpha-adrenergic binding site for (3H)WB-4101 in rat hippocampus 18 days postlesion without altering the specific binding of (3H)5-HT to serotonergic receptors. The chronic i.c.v. infusion of serotonin by minipump started at the appropriate time averted or reverted the effect. The dynamics of noradrenergic neurotransmission in the hippocampus was not impaired by lesion of the median raphe nucleus as determined by the uptake and turnover of noradrenaline as well as its release - as reflected by the normetanephrine concentration. In addition, neurotoxin-induced lesion of the dorsal noradrenergic bundle failed to alter either the Bmax or the Kd of (3H)WB-4101 binding to the nM site. Kainic acid-induced destruction of perikarya depressed the nM (3H)WB-4101 binding sites by 60% and completely prevented the up regulation caused by lesion of the median raphe nucleus. Thus, the supersensitivity-like response of the adrenoceptors to the lack of serotonin appears to be localized on kainate-sensitive cells within the hippocampus. PMID- 2991684 TI - The anticonvulsant effects of DADLE are primarily mediated by activation of delta opioid receptors: interactions between delta and mu receptor antagonists. AB - Dose-response comparisons of the ability of the selective delta antagonist ICI 154,129 (12.5-50 nmol), the nonselective antagonist naloxone (29-290 nmol), and the irreversible selective mu antagonist beta-fNA (1.3-21 nmol) to alter the threshold response to DADLE or etorphine was studied in the rat flurothyl seizure test. DADLE (35 nmol, i.c.v.) and etorphine (122 nmol/kg, s.c.) both caused increases in seizure threshold which were differentially antagonized by pretreatment (i.c.v.) with the respective antagonists. For DADLE, only ICI 154,129 and naloxone produced a dose-related blockade of the increase in seizure threshold, with ICI 154,129 being more potent than naloxone. In contrast, the anticonvulsant action of etorphine was not antagonized by ICI 154,129 (50 nmol), but was blocked by a low dose of naloxone (29 nmol) or beta-fNA (21 nmol). In addition, prior occupancy of mu-sites with beta-fNA (21 nmol) significantly diminished the abilities of either ICI 154, 129 (50 nmol) or naloxone (290 nmol) to antagonize the anticonvulsant action of DADLE. The results of this study demonstrated that the effects of DADLE to increase seizure threshold in the rat were primarily mediated by activation of a delta-opioid receptor system. Furthermore, evidence has been provided for a functional interaction between delta and mu receptors in the opioid regulation of seizure threshold. PMID- 2991685 TI - Adrenergic receptors on cerebral microvessels in control and parkinsonian subjects. AB - The binding of adrenergic ligands (3H-prazosin, 3H-clonidine, 3H dihydroalprenolol) was studied on a preparation of cerebral microvessels in the prefrontal cortex and putamen of control and Parkinsonian subjects. The adrenergic receptor density in microvessels of control patients was less than 0.5% and 3.3% respectively of the total binding. A significant decrease in the number of alpha-1 binding sites was observed on microvessels in the putamen of patients with Parkinson's disease. PMID- 2991686 TI - Increased gluconeogenesis in the rat at term gestation. AB - In this study the contribution of maternal gluconeogenesis to the glucose homeostasis of the maternal-fetal unit has been studied in fed term pregnant rats. We have measured the activity of two gluconeogenic enzymes, the rates of lactate turnover and the rates of gluconeogenesis from lactate in fed term pregnant rats. A decrease in plasma glucose and liver glycogen concentrations, and an increase of plasma lactate and alanine concentrations were observed in fed 22-day pregnant rats compared to virgin controls. Also, liver and kidney phosphoenolpyruvate carboxykinase activities and liver lactate dehydrogenase and hexose bisphosphatase activities significantly increased in fed term pregnant rats compared to virgin rats. The lactate turnover rate and the rate of gluconeogenesis in vivo from L-[U14C] Lactate increased four- and two-fold respectively in fed pregnant rats compared to fed virgins. PMID- 2991687 TI - Increased digitalis-like activity in human cerebrospinal fluid after expansion of the extracellular fluid volume. AB - The present study was designed to determine whether acute expansion of the extracellular fluid volume influenced the digitalis-like activity of human cerebrospinal fluid (CSF), previously described by our laboratory. Human CSF samples, drawn before and 30 minutes after the intravenous infusion of 1 liter of either saline or glucose solutions, were assayed for digitalis-like activity by inhibition of either the 86Rb+ uptake into human erythrocytes or by the activity of a purified Na+ - K+ ATPase. The CSF inhibitory activity on both systems significantly increased after the infusion of sodium solutions but did not change after the infusion of glucose. These results indicate that the digitalis-like factor of human CSF might be involved in the regulation of the extracellular fluid volume and electrolyte content and thereby in some of the physiological responses to sodium loading. PMID- 2991688 TI - A novel inhibitor of mammalian collagenase. AB - N-[[[(5-chloro-2-benzothiazolyl)thiolphenyllacetyll-L-cysteine (WY-45,368) is a potent inhibitor of human skin fibroblast collagenase. Kinetic data show that the inhibition is competitive, with a Ki of 3.5 microM. WY-45,368 inhibits neither of two other metalloproteinases, thermolysin and angiotensin converting enzyme, nor does it inhibit clostridial collagenase--thus indicating specificity for mammalian collagenase. PMID- 2991690 TI - Stimulation of cyclic GMP formation in smooth muscle cells by atriopeptin II. AB - Addition of synthesized atriopeptin II (AP-2), a 23 amino acid peptide of rat atria, to rat thoracic aorta smooth muscle cells results in the stimulation of cyclic GMP production by the cells. The EC50 for the effect is 81 nM and a 7 fold increase occurs at 10 microM AP-2. Cyclic GMP levels increased within 15 seconds after the addition of AP-2 and were maximal at 5 minutes. Cyclic GMP levels in primary rabbit kidney cells were increased 15 fold by 10 microM AP-2. However, no increase in cyclic GMP was detected in WI-38 fibroblast cells after the addition of 10 microM AP-2. Cyclic AMP levels were not affected by AP-2 in any of these cell systems. The effect upon cyclic GMP accumulation was specific for AP-2; none of the other compounds or peptides tested affected cyclic GMP levels. PMID- 2991689 TI - The effect of ACTH4-9 analog (Org2766) on some cerebrospinal fluid parameters in patients with Alzheimer's disease. AB - In a double-blind, placebo-controlled study, the central nervous system effects of an ACTH4-9 analog, Org2766 (40 mg/day), in Alzheimer's disease were assessed by measuring cerebrospinal fluid parameters during 6 months' treatment. Somatostatin-like immunoreactivity and cholinesterase activity, which are known to be reduced in cerebrospinal fluid of Alzheimer patients compared with controls, did not change during treatment. As a marker of noradrenergic and dopaminergic systems, we measured dopamine-beta-hydroxylase and homovanillic acid, but both levels were static. These results suggest that Org2766 did not interact with the transmitter systems, which are thought to be disturbed in Alzheimer's disease. PMID- 2991691 TI - The methylenedioxyindene calcium antagonists. PMID- 2991692 TI - In ability of Na+,K+-ATPase inhibitor to cause hypertension in sodium-loaded or deoxycorticosterone-treated one kidney rats. AB - The role of an endogenous inhibitor of Na+,K+-ATPase in hypertension observed in one-kidney NaCl-loaded rats treated with deoxycorticosterone (DOC) was examined. Ouabain or digitoxin, an exogenous inhibitor of Na+,K+-ATPase, failed to cause hypertension in one-kidney NaCl-loaded rats without DOC treatment or one-kidney DOC-treated rats without NaCl loading. Moreover, neither ouabain nor digitoxin acted additively with a putative endogenous inhibitor of Na+,K+-ATPase to augment hypertension observed in one-kidney NaCl-loaded rats treated with DOC. The results do not support the hypothesis that an endogenous inhibitor of Na+,K+ ATPase plays an important role in the development or maintenance of hypertension in this animal model. PMID- 2991694 TI - [Analysis of the effectiveness of repeated radiation treatment of lung cancer]. AB - In spite of a direct effect obtained in radiotherapy of lung cancer, tumor growth often resumes afterwards, metastases appear, all these causing death of most patients within the first 1-2 yrs. It is worth nothing that a considerable number of patients die from tumor recurrences in the absence of generalization. Therefore a question arises as to the therapeutic tactics for such patients, including the potentialities and appropriateness of repeated irradiation. The efficacy of repeated radiation therapy of 100 patients with lung cancer was analysed. Direct improvement was noted in 44% of x-ray findings and in 77% of clinical findings. The patients' mean survival rates were 29.7-2.6 mos. after the 1st course of treatment and 17.2 +/- 2.5 mos. after repeated irradiation. Short- and longterm results showed correlation, i.e. in x-ray improvement the mean survival after the 2nd course was 23.5 +/- 4 mos. whereas in its absence it was 13.5 +/- 4 mos. It was established that the longer the interval between the therapeutic courses the more probable an increase in a survival period after repeated radiation exposure. After 2 radiotherapy courses all the patients developed radiation pneumosclerosis of varying degree pathological rib fractures were marked in 4 patients and tumor regression in 5 patients. A conclusion has been made that repeated radiation therapy of lung cancer patients is of palliative nature. PMID- 2991693 TI - Differential effects of lipoxygenase products on FMLP and LTB4 evoked neutrophil aggregation. AB - There is evidence that the endogenous biosynthesis of LTB4 is involved in the aggregation of human neutrophils induced by the chemotactic peptide f-met-leu-phe (FMLP). If LTB4 mediates this aggregatory response, then agents which desensitize neutrophils to LTB4 should inhibit the cellular response to FMLP. Since many lipoxygenase products modulate other neutrophil responses to LTB4 and FMLP, we have investigated the effects of lipoxygenase products on LTB4- and FMLP initiated aggregation. Prior exposure to low concentrations of LTB4 (0.5-10 nM) inhibited subsequent aggregation to the same agent (50 nM), but it did not influence the response to FMLP (10(-7) M). Relatively high concentrations of 5 HETE (5-50 microM) inhibited aggregation initiated by either stimulus. Although the hydroperoxy derivative 5-HPETE also inhibited the response to LTB4, in the relatively narrow concentration range of 1-4 microM it stimulated FMLP-induced aggregation. This latter effect was confirmed using 12 cell preparations from six separate donors; it (the activity of 5-HPETE) was not mimicked by other 5 lipoxygenase products, including LTB4, nor the dihydroperoxide 8,15-DiHPETE. Our results indicate that neutrophil aggregation in response to LTB4 or FMLP can be selectively potentiated or inhibited. On the basis of these data we conclude that the endogenous synthesis of LTB4 is not directly involved in the neutrophil aggregatory response to FMLP, although the hydroperoxy intermediate 5-HPETE may act to enhance the cellular response. PMID- 2991695 TI - [Lymphography in cancer of the prostate]. AB - The frequency and area of lymphogenic metastasis with relation to the histological structure, differentiation and degree of local dissemination of a primary tumor were determined at lymphography of 114 patients with prostatic cancer. Lymph node involvement was often observed in poorly differentiated and solid trabecular adenocarcinoma, scirrhous cancer and locally disseminated tumors extending beyond the prostatic capsule. Metastases were usually located in the external and common inguinal lymph nodes. Lymphography was shown to be indicated to patients with a local spread of prostatic cancer (T2, T3 and T4). PMID- 2991696 TI - [Scintigraphy of focal brain lesions]. PMID- 2991697 TI - [Radionuclide study of reparative osteogenesis]. PMID- 2991698 TI - [Hormone levels of the blood of the adrenal veins and inferior vena cava in patients with adrenal and kidney diseases]. AB - Hydrocortisone and aldosterone concentration was determined in 138 patients with arterial hypertension of adrenal and renal genesis in the blood of the adrenal veins and in the cistern of the vena cava inferior, that of deoxycorticosterone in 50 patients, ACTH in 51 and renin activity in the blood plasma of the renal veins in 21 patients. The concentration of steroid hormones adequately reflected adrenal cortex function facilitating differential diagnosis between renal and adrenal pathology variants. Differential diagnostic analysis on the basis of change in the concentration of steroid hormones was found difficult or impossible if the patients received steroidogenesis changing drugs on day, preceding veno graphic examination of the adrenals. PMID- 2991699 TI - [Selective effect of isoproterenol on radiosensitivity of cells associated with differences in beta-adrenoreceptor expression]. AB - During x-ray irradiation of the joint cultures of cells 431 and cells B-82 of fibroblasts the antiradiation effect of isoproterenol manifests itself in regard to cells 431 only. As distinct from cells B-82, they bear beta-adrenoreceptors. The effect on them results in an increase in the cAMP intracellular content mediating the radioprotective effect. It has been noted that mammalian cells in malignancy often lose receptors to some hormones and biologically active substances which produce a radio-protective effect. It is assumed that the loss of receptors by tumor cells may be a factor opening up a new opportunity to the protection of normal cells from radiation injury in cancer radiotherapy. PMID- 2991700 TI - Response of serum hormones to androgen administration in power athletes. AB - Endocrine effects of self-administration of high doses of anabolic steroids and testosterone were investigated in five power athletes during 26 wk of training, and for the following 12-16 wk after drug withdrawal. After 26 wk of anabolic steroid and testosterone administration, serum testosterone concentrations had increased 2.3-fold. This was associated with increased concentrations of serum estradiol, which rose 7-fold to values (0.48 nmol X 1(-1) typical for females. There was a major decrease in serum FSH and LH concentrations, but they returned to control levels following drug withdrawal. However, serum testosterone concentrations stayed at low levels (9 nmol X 1(-1) ) during this follow-up period, indicating long-lasting impairment of testicular endocrine function. Serum ACTH concentrations were also decreased during steroid administration, possibly due to a corticoid-like effect of some of the anabolic steroids taken in high doses. However, no changes were seen in serum cortisol. The only consistent change in the control group was an increase in serum LH concentrations during the most intensive training, suggesting that a decreasing tendency of serum testosterone was compensated for by augmented LH secretion. PMID- 2991701 TI - [Nutritional status and food consumption in the north of the Ivory Coast]. AB - Malnutrition is frequent in the north of Ivory Coast, catching 63,7% of the children admitted in the Korhogo hospital, 39% of the children in the out patients services, 25% of the children living in villages and 6,9% of the admitted adults. We note animal protein and lipid deficiencies mainly. Syndrome of protein deficiency is very well known, but lipid deficiency is less; its responsibility in peripheral neuropathy, as hypothesis, is recalled. Fish and butcher's meat protein consumption remains stable, because of the economic context. Hunting meat consumption remains stable and important. Lipid intake is also under economic and cultural constraints. Development projects, and difficulties are briefly exposed. PMID- 2991702 TI - Synthesis of allylic and homoallylic isoprenoid pyrophosphates. PMID- 2991703 TI - Enzymatic hydrolysis of polyprenyl pyrophosphates. PMID- 2991704 TI - Purification and characterization of undecaprenylpyrophosphate synthetase. PMID- 2991706 TI - Mevalonate kinase. PMID- 2991705 TI - Radioenzymatic assay of plasma mevalonate. PMID- 2991707 TI - Phosphomevalonate kinase from pig liver. PMID- 2991708 TI - Mammographic breast cancer screening--a randomized trial in Malmo, Sweden. AB - A randomized trial is presented on the effect of repeated invitation to breast cancer screening with mammography on mortality from breast cancer. The invited group and the control group each consisted of approximately 21,000 women aged 45 69 yr at the start of the screening. The attendance rate was 74% at the first screening and 70% at the two subsequent screening rounds. The cancer detection rate was 7.5 per 1000 women examined in the first screening round and 2.2 and 2.0 per 1000 woman-years in the second and third screenings with an incidence of 0.9 in the intervals. The incidence in the control group was 2.7 per 1000 woman years. The proportion of positive biopsies was 61% in the first screening round, 33% in the second, and 58% in the third. After the prevalence screening, the stage distribution was more favourable in the invited group (including non attenders) than in the control group. In the two most recent periods of the programme, 62 out of 160 women with cancer (39%) in the invited group were in stage II-IV compared with 91 out of 159 (57%) in the control group. More than 60% of cancers detected at screening were either non-invasive or invasive with a diameter of 1 cm. The corresponding percentage in the control group was 27%. The importance of sampling bias is discussed. Although data on mortality still have to be awaited, the results so far clearly indicate a positive effect of screening. PMID- 2991710 TI - Drug-resistant mutants of herpes simplex virus type 2 with a mutator or antimutator phenotype. PMID- 2991709 TI - Steatitis in mice infected with Coxsackie virus B3. PMID- 2991711 TI - Cytomegalovirus infection. PMID- 2991712 TI - False-positive anti-HTLV-III assay results and HLA-DR4. PMID- 2991713 TI - Alternative medicine. PMID- 2991714 TI - Alternative vs orthodox medicine. PMID- 2991715 TI - [Rotavirus infections in newborn infants]. PMID- 2991716 TI - [Usefulness of isotopic cystography in the study of vesicoureteral reflux in children]. PMID- 2991717 TI - Pathophysiology of neuronal signaling in the cerebral cortex. PMID- 2991718 TI - [Progonoma or neuroectodermal melanotic tumor of infancy (NMTI)]. PMID- 2991720 TI - Marketing your services. PMID- 2991721 TI - Results of nursing ethics survey: allocation of scarce resources identified as major concern. PMID- 2991719 TI - [Use of calcium hydroxyapatite in oromaxillofacial surgery. I. General properties and the histochemical and histophysical evaluation of calcium hydroxyapatite]. PMID- 2991722 TI - Identification and reporting child abuse. PMID- 2991723 TI - Advocating for chemically dependent nurses--a job for the RLP. PMID- 2991724 TI - NLRB decision revokes NY unit certification. PMID- 2991725 TI - The morbidity and mortality of brain tumors. A perspective on recent advances in therapy. AB - This article reviews current morbidity and mortality statistics for the major classes of primary brain tumors including malignant astrocytoma, glioblastoma, low-grade astrocytoma, oligodendroglioma, meningioma, and other benign tumors and metastatic tumors. Innovations in therapy are discussed for surgery, radiation, chemotherapy, and such newer areas as hyperthermia, immunotherapy, and phototherapy. PMID- 2991726 TI - Aneurysms and arteriovenous malformations. AB - This article discusses intracranial aneurysms and subarachnoid hemorrhage in terms of prevalence, etiology, pathology, pathophysiology, and clinical presentation. Evaluation of patients, medical management, and surgical treatment are detailed as are results for selected aneurysms. PMID- 2991727 TI - Peripheral nerve surgery. AB - In treating the three main surgical problems of peripheral nerves--nerve sheath tumors, entrapment neuropathies, and acute nerve injuries--the overriding consideration is the preservation and restoration of neurologic function. Because of this, certain other principles may need to be compromised. These include achieving a gross total excision of benign tumors, employing conservative therapy as long as a disease process is not clearly progressing, and delaying repair of a nerve transection until the skin wound has healed. Only three pathophysiologic processes need be considered: neurapraxia (focal segmental dymyelination), axonotmesis (wallerian degeneration caused by a lesion that does not disrupt fascicles of nerve fibers), and neurotmesis (wallerian degeneration caused by a lesion that interrupts fascicles). With nerve sheath tumors and entrapment neuropathies, the goal is minimize the extent to which neurapraxia progresses to axonotmesis. The compressive force is relieved without carrying out internal neurolysis, a procedure that is poorly tolerated, presumably because a degree of nerve ischemia exists with any long-standing compression. When the nerve has sustained blunt trauma (through acute compression, percussion, or traction), the result can be a total loss of function and an extensive neuroma-in-continuity (scarring within the nerve). However, the neural pathophysiology may amount to nothing more than axonotmesis. Although this lesion, in time, leads to full and spontaneous recovery, it must be differentiated from the neuroma-in-continuity that contains disrupted fascicles requiring surgery. Finally, with open nerve transection, the priority is to match the fascicles of the proximal stump with those of the distal stump, a goal that is best achieved if primary neurorrhaphy is carried out. PMID- 2991728 TI - Update: Public Health Service workshop on human T-lymphotropic virus type III antibody testing--United States. PMID- 2991729 TI - Isolation of human T-lymphotropic virus type III/lymphadenopathy-associated virus from serum proteins given to cancer patients--Bahamas. PMID- 2991730 TI - Recommendations for preventing possible transmission of human T-lymphotropic virus type III/lymphadenopathy-associated virus from tears. PMID- 2991731 TI - [Hepatectomy after transcatheter arterial embolization (TAE) for hepatocellular carcinomas]. AB - Subjects of this study were 26 patients with hepatocellular carcinoma who underwent hepatectomy after TAE. They did not develop serious complications due to TAE. A suitable interval between TAE and the planned hepatectomy was about one month, based on AFP levels and the recovery of liver functions. Hepatic arteriography after TAE was useful to observe changes of the blood flow into the liver and the carcinoma, as well as to find intrahepatic metastases. It is also necessary for making final decisions on the strategy of the hepatectomy. The effects of TAE greatly depended on patterns of arterial blood feeding. When the tumor was less than 5 cm in diameter, had a capsule, and was fed by a single artery the necrotic rate was high. TAE was effective in intrahepatic metastases 0.5 cm or more in diameter, which were detectable by hepatic arteriography. TAE had little effect on intrahepatic metastases less than 0.5 cm in diameter, on intracapsular or extracapsular invasion, or on tumor embolus. For tumor embolus in the portal vein, since TAE and hepatectomy give poor results, other methods should be considered. PMID- 2991732 TI - Evidence for the epsilon-type of opioid receptor in the rat vas deferens. AB - The epsilon-opioid receptor type has been postulated to be specific for beta endorphin on the basis of structure-activity studies in the isolated rat vas deferens (RVD). Since beta-endorphin also displays high affinity for other opioid receptors, the present investigations were conducted to better define the epsilon receptor in the RVD. Therefore, the interaction of concomitantly acting pairs of opioid agonists was examined and analyzed according to the predictions of the law of mass action and the receptor theory. Our data provide evidence for a specific receptor for beta-endorphin in this organ, a receptor not recognized by either the delta-agonist D-Ala2-D-Leu5-enkephalin, or the mu-agonists FK-33824, fentanyl, and etorphine in the range of concentrations necessary for them to exert maximal inhibition of electrically induced twitch tension. The activity displayed by beta-endorphin in the RVD is in agreement with the existence of the proposed epsilon-type of opioid receptor therein. PMID- 2991733 TI - Thymidylate synthase gene amplification in fluorodeoxyuridine-resistant mouse cell lines. AB - We have previously isolated fluorodeoxyuridine-resistant mouse fibroblast (LU3-7) and neuroblastoma (FUdR-R) cell lines that overproduce thymidylate synthase and the mRNA for this enzyme up to 50-fold as compared to the parental cell lines. We have also cloned cDNA corresponding to mouse thymidylate synthase mRNA into pBR322. In the present study, we used this cloned cDNA as a hybridization probe in Southern blot analysis of DNA from the parental and overproducing cell lines. These analyses showed that the thymidylate synthase gene is amplified 50-100 fold in LU3-7 cells and about 30-fold in FUdR-R cells when compared to the respective parental cells. The sizes of the restriction fragments were the same in the parental and overproducing cells of each type, suggesting that extensive rearrangements have not occurred in the vicinity of the thymidylate synthase gene during the amplification process. However, not all of the fragments in the parental cells were amplified in the overproducing cells, suggesting that there may be multiple genes or pseudogenes for the enzyme. Restriction fragment length polymorphisms were detected when analyzing DNA from several different mouse cell lines. When LU3-7 cells were grown in the absence of selective pressure, the level of thymidylate synthase overproduction and the number of copies of the thymidylate synthase gene decreased in parallel. PMID- 2991734 TI - Expression of both alpha 1- and alpha 2-adrenoceptors in an insulin-secreting cell line. Parallel studies of cytosolic free Ca2+ and insulin release. AB - Changes in intracellular free Ca2+, [Ca2+]i, and immunoreactive insulin release in response to alpha-adrenergic agents were measured in RINm5F cell suspensions. Cells were loaded with the fluorescent indicator quin 2 for monitoring [Ca2+]i. Epinephrine (1 microM), which inhibited alanine-stimulated insulin release by 73%, evoked a transient rise in [Ca2+]i. This rise is in part due to Ca2+ mobilization, since it is still present in the absence of extracellular Ca2+. The alpha 2-adrenergic agonist clonidine (10 microM) mimicked the epinephrine effect on insulin release without any change in [Ca2+]i. In contrast, the alpha 1 adrenergic agonist phenylephrine (10 and 100 microM) raised [Ca2+]i, albeit to a lesser extent than epinephrine. Phenylephrine enhanced basal, but had no effect on alanine-stimulated insulin release. To examine further the nature of the effect of epinephrine, specific alpha-adrenergic blocking agents were employed. The epinephrine-induced increase in [Ca2+]i could be inhibited by the alpha 1 adrenergic antagonists BE2254 (0.1 microM) and prazosin (0.01 microM). In the presence of these blockers, epinephrine was still able to inhibit insulin release. When alpha 2-adrenergic receptors were blocked by the addition of idazoxan (0.1 and 1 microM), epinephrine still raised [Ca2+]i. At the higher concentration of idazoxan, the epinephrine inhibition of insulin release was completely overcome. The alpha-adrenergic agonists did not attenuate the alanine induced rise in [Ca2+]i. This study shows that both subtypes of alpha-adrenergic receptors are present in the insulin-secreting cell line RINm5F. The activation of alpha 1-adrenergic receptors leads to an increase in [Ca2+]i. In contrast, the inhibition of insulin release due to epinephrine is mediated through alpha 2 adrenergic receptors. The alpha 2-adrenergic mechanism does not involve changes in [Ca2+]i, but is rather exerted at a later step in the secretory process. PMID- 2991735 TI - Multiple states of opioid receptors may modulate adenylate cyclase in intact neuroblastoma X glioma hybrid cells. AB - Opioid receptor binding and opioid-mediated inhibition of cAMP accumulation were studied simultaneously in intact NG108-15 cells. The dose-response curves for the biological response were suggestive of positive cooperativity and systematically occurred at lower ligand concentrations than those for the binding of [3H] [D Ala2, D-Leu5]enkephalin (DADLE), which were instead shallow and suggestive of a site heterogeneity or of a cooperative phenomenon. Computer modeling of the binding isotherms revealed that the data are best described assuming two binding sites with different affinities for the agonist; the mean ratio between the DADLE concentrations yielding half-maximal occupancy of the high affinity site and half maximal response was 1.5, but it was 36 when the fractional occupancy of the sum of the two sites was considered. On examining several opioids, no direct correlation was found between high affinity site and biological response; however, several agonists displayed different affinities for the two sites, while the antagonist naloxone and the partial agonist diprenorphine bound to them with identical affinities. Furthermore, naloxone exhibited a good agreement between half-maximal receptor occupancy and Ki in blocking the agonist response. Thus, the binding heterogeneity detectable in intact cells is agonist-specific, and suggests rather that the sites are states of an identical receptor population. When [3H]diprenorphine was used to label the opioid receptors, the competition curves for DADLE were consistent with the existence of an additional, very low affinity state undetectable by direct binding with labeled agonist and, again, not discriminated by naloxone. Multiple affinity states of the opioid receptor in intact cells may reflect its interaction with the effector system in the plasma membrane. PMID- 2991736 TI - Binding of an 125I-labeled thromboxane A2/prostaglandin H2 receptor antagonist to washed canine platelets. AB - A binding site for the thromboxane A2/prostaglandin H2 (TXA2/PGH2) antagonist 125I-PTA-OH (9,11-dimethylmethano-11,12-methano-16-(4-methoxyphenyl)-13,14-dih ydro-13-aza-1 5 alpha beta-w-tetranor-TXA2) to washed canine platelets is described. 127I-PTA-OH competitively antagonized aggregation induced by the TXA2/PGH2 mimetic U46619. A Schild analysis of the pharmacologic study revealed pA2 of 7.97 and a slope of -0.95. The pA2 value yielded a Kd of 11 nM. Specific binding in Tris-NaCl buffer (pH 7.4) is not affected by extracellular Ca2+ or Mg2+ in concentrations up to 750 microM. The pH optimum for binding resides between 7.0 and 7.4. The association rate constant, k1, was 4.5 X 10(6) M-1 min 1, and the dissociation rate constant, k-1, was 1.45 X 10(-1) min-1, yielding a kinetically determined Kd (k-1/k1) of 32 nM. Scatchard analysis of I-PTA-OH binding to washed canine platelets revealed two classes of binding sites, a high affinity site (Kd = 24 nM, Bmax = 71 fmol/10(7) platelets) (4400 binding sites/platelet) and a low affinity site (Kd = 2.1 microM). Several TXA2/PGH2 receptor antagonists competed with specific 125I-PTA-OH binding, and the rank order of potency for displacing the ligand correlated (r = 0.97) with the rank order of potency for their ability to inhibit U46619-induced aggregation in canine platelet-rich plasma. Prostaglandins F2 alpha and E2 also displaced the ligand, but only at much higher concentrations. Binding of I-PTA-OH or the TXA2/PGH2 mimetic U46619 was unaffected by the aggregating agents epinephrine (10 microM) or ADP (5 microM). The similarity in the Kd values obtained kinetically, by equilibrium binding studies for the high affinity site and by Schild analysis, suggests that this high affinity site mediates TXA2/PGH2 induced platelet aggregation. In addition, the close correlation between the abilities of the antagonists to displace the ligand and to inhibit U46619-induced aggregation suggests that this site may represent a TXA2/PGH2 receptor. PMID- 2991737 TI - Adenosine-induced K+ current in Xenopus oocyte and the role of adenosine 3',5' monophosphate. AB - Voltage clamp technique was used in Xenopus laevis oocytes in order to study and compare membrane currents evoked by extracellularly applied adenosine (0.1-10 microM) and intracellularly injected cyclic AMP (0.15-10 microM). The adenosine response is a late long-lasting outward K+ current ("H" current), mediated by the Ra purine receptor subtype. The H current amplitude is directly proportional to (occupancy)3; the KD for adenosine is 3.34 microM. The H current is inhibited by the intracellular injection of protein kinase inhibitors, types II and III (5-450 ng/oocyte) and is usually potentiated by intracellular injection of theophylline (100-300 microM), though extracellular application of theophylline (1-100 microM) reversibly blocks the receptor. Occasionally, the H current is contaminated by a small Cl- current. The cyclic AMP current is also a long-lasting K+ outward current which is potentiated by extracellular theophylline (2 mM). Injection of cyclic AMP inhibits the membrane response to subsequent application of adenosine. The converse inhibition of a cyclic AMP response by an earlier adenosine response is also observed but at very high concentrations of adenosine (greater than 0.6 mM). It was shown by radioimmunoassay that extracellular adenosine increases the level of the intracellular cAMP within a few seconds by about 30%. Intracellular injection of a comparable amount of cAMP was shown to evoke a measurable K+ current. It is proposed that the adenosine-evoked K+ outward current is mediated by a rise in intracellular cAMP. PMID- 2991738 TI - Molecular structure of Ro15-1788 and a model for the binding of benzodiazepine receptor ligands. Structural identification of common features in antagonists. AB - Ligands that bind to the benzodiazepine receptor have three possible effects. The ligand can be an agonist and reduce anxiety, an antagonist and have no biological effect, or an inverse agonist and promote convulsions. This receptor complex is unique in its spectrum of response to ligands, and conformational changes in the receptor are implicated. The x-ray crystal structure of an imidazobenzodiazepine antagonist ligand, Ro15-1788, was determined and compared to the structures of the 1,4-benzodiazepine agonists and to two other types of antagonists, beta carbolines and a pyrazoloquinolinone, CGS-8216. The antagonists were found to have similar arrangements of binding features including an aromatic ring, a carbonyl oxygen atom, and a hydrophobic side chain. The structures of these antagonists could be superimposed in a model binding site with three common features for all of the antagonists and a fourth hydrogen-bonding site for the pure antagonists (or inverse agonists), the beta-carbolines, and CGS-8216. A comparison of the shapes of the antagonist benzodiazepine, Ro15-1788, and several agonists showed that RO15-1788 has a unique azepine ring conformation that distorts the usual arrangement of the aromatic A ring, carbonyl oxygen atom, and imine N atom of the agonists. A conformational adjustment in the receptor would be required to accommodate both of these types of ligands. A summary of the superpositions of typical agonists and the antagonists leads to a model with 7 conformationally mobile binding points. Inverse agonists are distinguished from antagonists by the length of the hydrophobic side chain. Antagonists are distinguished from agonists in part by the lack of a binding feature similar to the imine N atom of the diazepine ring. This model accounts for the key features found in ligands for the benzodiazepine receptor and provides an explanation for the spectrum of responses elicited by receptor binding. PMID- 2991739 TI - Specificity of inhibitors of poly(ADP-ribose) synthesis. Effects on nucleotide metabolism in cultured cells. AB - The effects of inhibitors of poly(ADP-ribose) synthesis on cell growth and several parameters of nucleotide metabolism have been determined. At concentrations which produced similar inhibitions of poly(ADP-ribose) synthesis, 3-acetylaminobenzamide (1 mM) had no effect on L1210 cell growth, 3 aminobenzamide (5mM) was slightly inhibitory and 3-methoxybenzamide (5 mM) was a potent inhibitor of growth. During a 2-h incubation, none of the inhibitors affected ribo- or deoxyribonucleotide concentrations in cells treated with or without N-methyl-N-nitrosourea; however, N-methyl-N-nitrosourea treatment reduced dCTP concentrations by 50%. During a 24-hr incubation, 3-aminobenzamide and 3 acetylaminobenzamide did not lower ribonucleotide concentrations in cells grown with either undialyzed or dialyzed serum. In contrast, 3-methoxybenzamide caused a depletion of UTP in cells grown with undialyzed serum and caused a depletion of all purine and pyrimidine ribonucleotides in cells grown with dialyzed serum. 3 Aminobenzamide and 3-acetylaminobenzamide had no effect on the conversion of hypoxanthine to ATP and GTP but did slightly inhibit incorporation of formate into ATP and GTP. 3-Methoxybenzamide inhibited incorporation of both hypoxanthine and formate into purine ribonucleotides. 3-Aminobenzamide, 3 acetylaminobenzamide, and 3-methoxybenzamide all inhibited glycine incorporation into ATP and GTP and reduced both the incorporation of thymidine into DNA and the apparent specific activity of the dTTP pool. We conclude that inhibition of poly(ADP-ribose) synthesis causes little or no growth inhibition and has no effect on purine or pyrimidine nucleotide synthesis de novo. The effect of all the inhibitors on glycine and formate metabolism may be related to an inhibition of ADP-ribose synthesis or may be a secondary effect of the inhibitors. The growth inhibition and the reduction in nucleotide concentration caused by 3 methoxybenzamide are apparently secondary effects of this drug and may result from an inhibition of phosphoribosyl pyrophosphate synthesis. PMID- 2991740 TI - Effects of pertussis toxin on cAMP and cGMP responses to carbamylcholine in N1E 115 neuroblastoma cells. AB - As noted previously, in N1E-115 neuroblastoma cells, carbamylcholine, a muscarinic cholinergic agonist, increased cGMP over 15-fold and decreased basal and prostaglandin E1 (PGE1)-stimulated cAMP content. In contrast to the stimulatory effects of PGE1 on cAMP, which were immediate, the carbamylcholine induced decrease in basal and PGE1-stimulated cAMP exhibited a delay. The delay in carbamylcholine inhibition was independent of the extent of adenylate cyclase activation. Although basal cAMP content was suppressed within 30 sec after addition of carbamylcholine, inhibition was not maximal for at least 2 min following agonist addition; the delay was similar in cells exposed to PGE1 for 10 min prior to carbamylcholine but could be eliminated by incubation of the cells with muscarinic cholinergic agonist for 5 min prior to addition of prostaglandin. N1E-115 neuroblastoma cells possess a 41,000-Da membrane protein believed to be a component of the inhibitory GTP-binding protein of adenylate cyclase that is ADP ribosylated by pertussis toxin. Incubation of the cells with pertussis toxin prior to the addition of carbamylcholine reduced the maximal extent of inhibition of cAMP content and prevented the [32P]ADP-ribosylation of a 41,000-Da protein by toxin and [32P]NAD in membrane preparations from these cells. Incubation of cells with pertussis toxin, however, did not significantly alter the dose-response curve for carbamylcholine effects on cGMP. Even high concentrations of carbamylcholine, effective in stimulating cGMP, had minimal effects on cAMP content in toxin-treated cells; thus, ADP-ribosylation of Gi converts the adenylate cyclase but not the guanylate cyclase system to an agonist-insensitive state. PMID- 2991741 TI - The relationships between receptor binding capacity for norepinephrine, angiotensin II, and vasopressin and release of inositol trisphosphate, Ca2+ mobilization, and phosphorylase activation in rat liver. AB - Concentration-response relationships for norepinephrine-, angiotensin II-, and vasopressin-stimulated changes in cell Ca2+ content, phosphorylase activation, and cytosolic free Ca2+ and myo-inositol-P3 levels were examined in isolated hepatocytes. The specific binding of radioligands to alpha 1-adrenergic, vasopressin, and angiotensin II receptors was also examined in rat liver plasma membranes. Disparities occurred between the concentration-response curves for myo inositol-P3 formation and the Ca2+ and phosphorylase responses, with the greatest difference being observed with vasopressin and the smallest with norepinephrine. It was also observed that all three agonists produced the same maximum changes in phosphorylase, cell Ca2+, and cytosolic Ca2+, but the maximum capacity of each agonist to generate myo-inositol-P3 varied greatly and was correlated with the maximum receptor binding capacity. The data indicated that a very small and submaximal elevation of myo-inositol-P3 was sufficient to maximally elevate cytosolic Ca2+ and activate phosphorylase. In addition, the relationship between the accumulation of myo-inositol-P3 and the elevation of cytosolic Ca2+ was similar, irrespective of whether the agonist was norepinephrine, angiotensin II, or vasopressin. It is proposed that the large differences between the concentration-response curves for myo-inositol-P3 formation and Ca2+ and phosphorylase changes observed with vasopressin and angiotensin II are due to the higher density of their receptors on liver cell plasma membranes compared with alpha 1-adrenergic receptors. PMID- 2991742 TI - Cyclic AMP, nuclear protein kinase and the PY815 cell cycle. AB - A substantial increase in cyclic AMP-dependent protein kinase activity occurred in nuclei of PY815 mastocytoma cells during G1 phase growth arrest by DB cyclic AMP and the increased nuclear protein kinase was accompanied by changes in nuclear protein phosphorylation. However, there was no obligatory association between the rise in nuclear cyclic AMP-dependent protein kinase in G1 phase and growth arrest because nuclear cyclic AMP-dependent protein kinase also increased during G1 phase in cycling PY815 cells synchronized with amethopterin. These observations suggest that maintenance of high cyclic AMP levels during G1 phase may cause growth arrest by activating a cyclic AMP-dependent protein kinase that normally increases in PY815 cell nuclei during G1 phase. PMID- 2991743 TI - Inhibition of the formation of lipid-linked intermediates in normal and transformed cells by a purified tunicamycin homologue. AB - The effects of a purified homologue of tunicamycin (B2-tunicamycin) on the biosynthesis of lipid-linked intermediates participating in protein glycosylation in normal embryonic fibroblasts, 3T3 and virally transformed (simian virus 40 and polyoma virus) mouse fibroblasts grown in culture were investigated. Long incubations (20 h) with the antibiotic caused a higher degree of inhibition of sugar incorporation into glycoproteins in transformed cells. However, the formation of lipid-linked intermediates was inhibited to a similar level in both cell types. When time dependent inhibition experiments were carried out using transformed cells, an earlier and stronger inhibition of the formation of lipid oligosaccharides occurred (70% inhibition at 30 min). In 3T3 cells, prolonged incubation (6-8 h) was necessary in order to reach a similar degree of inhibition. Formation of lipid-sugar was also inhibited to a greater extent by B2 tunicamycin in transformed cells. This inhibition was not clearly time dependent. Analysis of the newly synthesized glycolipids in 3T3 and in transformed cells after B2-tunicamycin treatment have shown reduction in dolichyl-P-P-sugars as well as in other glycolipids. Dimethylsulfoxide (10%) and linoleic acid (0.5 mg/ml) markedly increased the level of tunicamycin activity in 3T3 cells while phosphatidylcholine (2 mg/ml) partially reversed it. The stronger and faster inhibition of the formation of lipid intermediates of the dolichyl-phosphate cycle caused by B2-tunicamycin in transformed cells, described here for the first time, may therefore be due to differences in penetration of the antibiotic into these cells. PMID- 2991744 TI - Isolation and characterization of the SPT2 gene, a negative regulator of Ty controlled yeast gene expression. AB - The his4-917 mutation of Saccharomyces cerevisiae results from the insertion of the Ty element Ty917 into the regulatory region of the HIS4 gene and renders the cell His-. The hist4-912 delta mutant, which carries a solo delta in the 5' noncoding region of HIS4, is His+ at 37 degrees C but His- at 23 degrees C. Both these mutations interfere with HIS4 expression at the transcriptional level. The His- phenotype of both insertion mutations is suppressed by mutations at the SPT2 locus. The product of the wild-type SPT2 gene apparently represses HIS4 transcription in these mutant strains; this repression is relieved when the SPT2 gene is destroyed by mutation. The repression of transcription by SPT2 presumably results from an interaction between the SPT2+ gene product and Ty or delta sequences. In this paper, we report the cloning and DNA sequence analysis of the wild-type SPT2 gene and show that the gene is capable of encoding a protein of 333 amino acids in length. In addition, we show that a dominant mutation of the SPT2 gene results from the generation of an ochre codon which is presumed to lead to a shortened SPT2 gene product. PMID- 2991745 TI - Properties of some monkey DNA sequences obtained by a procedure that enriches for DNA replication origins. AB - Twelve clones of monkey DNA obtained by a procedure that enriches 10(3)- to 10(4) fold for nascent sequences activated early in S phase (G. Kaufmann, M. Zannis Hadjopoulos, and R. G. Martin, Mol. Cell. Biol. 5:721-727, 1985) have been examined. Only 2 of the 12 ors sequences (origin-enriched sequences) are unique (ors1 and ors8). Three contain the highly reiterated Alu family (ors3, ors9, and ors11). One contains the highly reiterated alpha-satellite family (ors12), but none contain the Kpn family. Those remaining contain middle repetitive sequences. Two examples of the same middle repetitive sequence were found (ors2 and ors6). Three of the middle repetitive sequences (the ors2-ors6 pair, ors5, and ors10) are moderately dispersed; one (ors4) is highly dispersed. The last, ors7, has been mapped to the bona fide replication origin of the D loop of mitochondrial DNA. Of the nine ors sequences tested, half possess snapback (intrachain reannealing) properties. PMID- 2991746 TI - Quantification of expression of linked cloned genes in a simian virus 40 transformed xeroderma pigmentosum cell line. AB - We wished to determine whether simian virus 40 (SV40)-transformed xeroderma pigmentosum cells, despite their defective DNA repair, were suitable for DNA mediated gene transfer experiments with linked genes. Expression of a nonselectable gene (cat, coding for chloramphenicol acetyltransferase [CAT]) linked to a selectable gene (gpt, coding for xanthine-guanine phosphoribosyltransferase [XPRT]) in the plasmid pSV2catSVgpt was quantified after transfection of SV40-transformed xeroderma pigmentosum [XP20s(SV40)] and normal human [GM0637(SV40)] fibroblast cell lines. A novel autoradiographic assay with [3H]xanthine incorporation showed 0.5 to 0.7% phenotypic expression of XPRT in both cell lines. Without selection, transient CAT activity was 20 times greater in the GM0637(SV40) than in the XP20s(SV40) cells, and transient XPRT activity was 5 times greater. Both of these transient activities were increased and equalized in both cell lines by transfection with pRSVcat or pRSVgpt. Genotypic transformation to gpt+ occurred at a frequency of 2 X 10(-4) to 4 X 10( 4) in both cell lines with pSV2catSVgpt. After 2 to 3 months in selective medium, stable expression of the (nonselected) cat gene was found in 11 (92%) of 12 gpt containing clones derived from GM0637(SV40) cells and in 13 (81%) of 16 gpt containing clones from XP20s(SV40) cells. However, the levels of CAT activity did not correlate with those of XPRT activity, and both of these activities varied more than 100-fold among different clones. Copies (1 to 4) of the gpt gene were integrated in four clones of the GM0637(SV40) cells having an XPRT activity of 1 to 5 nmol/min per mg, but 5 to 80 copies were integrated in four XP20s(SV40) clones with an XPRT activity of 0.8 to 1.8 nmol/min per mg. This study shows that XP20s(SV40) is as suitable for gene transfer experiments as the normal human line GM0637(SV40). PMID- 2991747 TI - Neoplastic transformation of normal and carcinogen-induced preneoplastic Syrian hamster embryo cells by the v-src oncogene. AB - The ability of cloned Rous sarcoma virus (RSV) DNA encoding the v-src oncogene to neoplastically transform normal, diploid Syrian hamster embryo (SHE) cells was examined. Transfection of RSV DNA into early passage SHE cells resulted in a low but significant number of tumors when treated cells were injected into nude mice. Tumors formed with a low frequency (two tumors out of ten sites injected) and only after a long latency period (14 weeks). In contrast to the normal SHE cells, several different carcinogen-induced preneoplastic immortal SHE cell lines were highly susceptible to transformation by the v-src oncogene to the neoplastic phenotype. Tumors formed with high efficiency and a short latency period (less than 3 weeks). Further studies were performed to determine the basis for the inefficient transformation of the normal SHE cells. NeoR clones isolated after cotransfection of SHE cells with pSV2-neo and RSV DNAs were neither morphologically altered nor immortal and did not contain detectable levels of the v-src gene product. These results suggest that neoplastic transformation by v-src DNA in the normal cells is initially suppressed. However, cells from a v-src induced tumor expressed v-src RNA, and antibody to v-src protein precipitated from the tumor cells a 60,000-molecular-weight protein which displayed protein kinase activity. Karyotypic analyses confirmed that the tumor was derived from Syrian hamster cells and suggested that it was clonal in nature. These results indicate that the v-src oncogene was primarily responsible for neoplastic transformation of SHE cells. In contrast to the results with the v-src oncogene, our previous studies showed that v-Ha-ras oncogene alone is unable to induce neoplastic transformation of SHE cells. Furthermore, the v-myc oncogene was able to compliment v-Ha-ras to neoplastically transform SHE cells, while cotransfection with v-src plus v-myc did not increase the incidence of tumors. PMID- 2991748 TI - Direct and indirect gene replacements in Aspergillus nidulans. AB - We performed three sets of experiments to determine whether cloned DNA fragments can be substituted for homologous regions of the Aspergillus nidulans genome by DNA-mediated transformation. A linear DNA fragment containing a heteromorphic trpC+ allele was used to transform a trpC- strain to trpC+. Blot analysis of DNA from the transformants showed that the heteromorphic allele had replaced the trpC allele in a minority of the strains. An A. nidulans trpC+ gene was inserted into the argB+ gene, and a linear DNA fragment containing the resultant null argB allele was used to transform a trpC- argB+ strain to trpC+. Approximately 30% of the transformants were simultaneously argB-. The null argB allele had replaced the wild-type allele in a majority of these strains. The A. nidulans SpoC1 C1-C gene was modified by removal of an internal restriction fragment and introduced into a trpC- strain by transformation with a circular plasmid. A transformant containing a tandem duplication of the C1-C region separated by plasmid DNA was self-fertilized, and trpC- progeny were selected. All of these had lost the introduced plasmid DNA sequences, whereas about half had retained the modified C1 C gene and lost the wild-type copy. Thus, it is possible with A. nidulans to replace chromosomal DNA sequences with DNA fragments that have been cloned and modified in vitro by using either one- or two-step procedures similar to those developed for Saccharomyces cerevisiae. PMID- 2991749 TI - Structure and localization of genes encoding aberrant and normal epidermal growth factor receptor RNAs from A431 human carcinoma cells. AB - A431 cells have an amplification of the epidermal growth factor (EGF) receptor gene, the cellular homolog of the v-erb B oncogene, and overproduce an aberrant 2.9-kilobase RNA that encodes a portion of the EGF receptor. A cDNA (pE15) for the aberrant RNA was cloned, sequenced, and used to analyze genomic DNA blots from A431 and normal cells. These data indicate that the aberrant RNA is created by a gene rearrangement within chromosome 7, resulting in a fusion of the 5' portion of the EGF receptor gene to an unidentified region of genomic DNA. The unidentified sequences are amplified to about the same degree (20- to 30-fold) as the EGF receptor sequences. In situ hybridization to chromosomes from normal cells and A431 cells show that both the EGF receptor gene and the unidentified DNA are localized to the p14-p12 region of chromosome 7. By using cDNA fragments to probe DNA blots from mouse-A431 somatic cell hybrids, the rearranged receptor gene was shown to be associated with translocation chromosome M4. PMID- 2991751 TI - Linear simian virus 40 DNA fragments exhibit a propensity for rolling-circle replication. AB - A linear simian virus 40 origin-containing DNA fragment replicated in monkey COS cells, generating tandemly repeated (head-to-tail) structures. Electron microscopy revealed circle-and-tail configurations characteristic of rolling circle replication intermediates. Circularization of the same DNA before transfection led to a theta type of replication which generated supercoiled DNA molecules. PMID- 2991750 TI - Erbb is linked to the alpha-globin locus on mouse chromosome 11. AB - A fragment of the human gene for c-erb-B was used to map homologous sequences in mice. Analysis of somatic cell hybrids and recombinant inbred and congenic mouse strains indicated that this gene, designated Erbb, is closely linked to the gene for alpha-globin on mouse chromosome 11. Several genes controlling hematopoietic differentiation map to mouse chromosome 11. PMID- 2991753 TI - Expression of transfected DNA in avian cells can be enhanced in trans by retroviral infection. AB - Using a quantitative S1 nuclease protection assay, we demonstrated that acute or chronic infection of avian cells enhances expression of an exogenously introduced rat preproinsulin II gene by approximately equal to 50-fold. The degree of enhancement is shown to vary with the transfection technique used but is independent of the transcription control region of the transfected gene. We conclude that retroviral infection of avian cells enhances expression of transfected DNA in trans by facilitating the uptake of DNA rather than by activating the transfected promoter. PMID- 2991752 TI - Large-scale production of polyoma middle T antigen by using genetically engineered tumors. AB - A recombinant plasmid containing a metallothionein promoter-polyoma middle T cDNA fusion was constructed and used to transfect NIH 3T3 cells. Transformed cells expressing middle T were injected into nude mice. Within 3 weeks, each mouse produced tumors containing middle T equivalent to that in 250 to 1,000 100-mm dishes of polyomavirus-infected cells. This middle T, partially purified by immunoaffinity chromatography, retained activity as measured by its ability to be phosphorylated in vitro. The combined approach of fusing strong promoters to genes of interest and utilizing nude mice to grow large quantities of cells expressing the gene provides a quick, inexpensive alternative to other expression systems. PMID- 2991754 TI - Spin labelling of sialic acid and galactose residues on lymphocyte plasma membrane: effects of lectins on oligosaccharide dynamics. AB - To investigate the behaviour of glycoprotein and glycolipid receptors at the lymphocyte cell surface, a spin label probe has been introduced into either sialic acid or galactose residues on lymphocyte plasma membrane, using specific activation of sugars with periodate or galactose oxidase, followed by reductive amination. The extent of membrane labelling could be controlled by varying the mole ratios of reactants used. Chloroform-methanol extraction of the labelled membranes showed that approximately 17% of the label is bound to glycolipids. A large fraction of the spin label could be released from both sialic acid and galactose-labelled membrane by treatment with pronase, indicating attachment to membrane proteins. Rotational correlation times (tau c) for both labelled sialic acid and galactose residues were in the range 10-13 X 10(-10) sec, indicating a reduction in sugar headgroup mobility at the membrane surface. Isolated lymphocyte membrane glycoproteins spin labelled on galactose residues and reassembled into phospholipid bilayer vesicles showed similar motional characteristics. Prolonged incubation of conc. suspensions of labelled membrane resulted in cleavage of the sialic acid-bound (but not the galactose-bound) label. Binding of several lectins to labelled plasma membrane produced significant immobilization of cell surface oligosaccharides while others had no effect. This differential restriction in oligosaccharide motion following lectin binding appears to be at least partly related to the sugar specificity of the lectin. Binding of wheat germ agglutinin and Ricinus communis agglutinin to sialic acid and galactose-labelled membrane respectively produced a dramatic decrease in oligosaccharide mobility which was reversible on addition of the appropriate sugar inhibitor. The concn dependence of lectin-induced spin label immobilization suggested a cooperative interaction between the lectins and their oligosaccharide receptors. Binding of lectins to the lymphocyte cell surface thus seems to have distinct effects on the dynamic state of glycoproteins and glycolipids within the glycocalyx. PMID- 2991756 TI - [Prenatally diagnosed ascites-causing cytomegalovirus infection in a fetus]. PMID- 2991757 TI - Current therapy of demyelinating neuropathies. PMID- 2991755 TI - Monoclonal antibody (EGR/G49) reactive with the epidermal growth factor receptor of A431 cells recognizes the blood group ALeb and ALey structures. AB - The carbohydrate specificity of the monoclonal antibody EGR/G49, raised against the epidermal growth factor (EGF) receptor of A431 cells, has been investigated by assessing its interactions with glycoproteins and erythrocytes derived from individuals of known blood group ABH, Lewis and secretor types, and by inhibition of binding assays using structurally defined oligosaccharides. The results indicate that this antibody reacts with the difucosylated blood group structures ALeb and ALey: (formula; see text) This antibody differs from the previously described anti-EGF receptor antibody. TL5, which is directed at the terminal blood group A trisaccharide structure and reacts poorly with the ALeb/Ley structures. Since both antibodies were selected for their reactivities with the receptor for EGF, their specificities provide evidence for the presence of both the mono- and difucosylated blood group A structures on the receptor glycoprotein. These antibodies will be invaluable in the studies of the distribution and the roles of blood group related carbohydrate structures in the organisation and function of the EGF and other receptor systems. PMID- 2991758 TI - Synergistic induction of sister-chromatid exchanges in lymphocytes from normal subjects and from patients under cytostatic therapy by inhibitors of poly(ADP ribose)polymerase and antitumour agents. AB - The effects of nicotinamide on SCE rates induced in vitro by chlorambucil (CBC or melphalan (MELPH) or mitomycin C (MMC) was studied. The combined treatments with either CBC or MELPH or MMC and nicotinamide showed the potentiating ability of the latter drug. Theophylline and MELPH were also found to act synergistically on the induction of SCEs. In a combined in vivo and in vitro study, lymphocytes taken from 7 cancer patients who had been given cytoxan by injection 3 h before, were treated with nicotinamide or diphylline (DP) in vitro, and found to have synergistically increased exchange rates. This has implications for interpreting the repair processes involved, for monitoring drug combinations that synergistically damage DNA in vivo and in vitro and for identifying interindividual variation in the response to the treatment. PMID- 2991759 TI - Scintillometric determination of unscheduled DNA synthesis in primary cultures of rat-liver cells. A hydroxylapatite batch assay. AB - A new procedure has been examined for measuring unscheduled DNA synthesis (UDS) in hepatocyte primary cultures by liquid-scintillation counting. DNA of the hepatocyte lysates was eluted with K-phosphate buffers after absorption on hydroxylapatite in order to reduce the background produced by cytoplasmic radioactivity. To inhibit hepatocyte replicative synthesis, hydroxyurea (10 mM) and cytosine arabinoside (80 microM) were added to the cultures. This procedure was found capable of detecting UDS elicited by 0.3 - 10 mM N nitrosodimethylamine. PMID- 2991760 TI - Enhanced induction of SV40 replication from transformed mammalian cells by fusion with UV-irradiated untransformed cells. AB - DNA-damaging agents such as ultraviolet (UV) light are known to cause stimulation of virus replication in SV40-transformed hamster and human cells. The dose response curves of UV-induced SV40 replication in transformed hamster cells resemble that obtained for UV-enhanced reactivation (ER) and UV-enhanced mutagenesis (EM) of SV40 or herpes viruses in mammalian cells. We have investigated whether UV-enhanced production of SV40 from transformed hamster (THK) and human (NB-E) cells belongs to the same category of conditional responses as ER and EM. To answer this question we have made use of the phenomenon that fusion of the SV40-transformed cells with monkey cells that are permissive to SV40 results in a considerable increase in the production of SV40 virus. When THK or NB-E cells were fused with UV-irradiated CV-1 cells at various times after irradiation, induction of SV40 was further increased and reached a maximum value of 2--3-fold when fusion was delayed for 24-48 h after irradiation. The kinetics of enhanced SV40 induction resembled that of ER and EM, suggesting that the UV-stimulated part of the induction represents one of the pleiotropic responses that are transiently induced in mammalian cells by DNA-damaging agents. Evidence is presented, showing that this transient effect can be induced only in cells that are permissive to SV40 replication. This suggests that the enhanced induction observed after fusion with irradiated monkey cells may be attributed to a transient increase in the activity of "permissiveness' factors. No enhanced induction was found when the THK or NB-E cells were fused with irradiated rodent cells, that are not or only slightly permissive to SV40 replication. PMID- 2991761 TI - Differential effect of ultraviolet light on the induction of simian virus 40 and a cellular mutator phenotype in transformed mammalian cells. AB - UV irradiation of simian virus 40 (SV40)-transformed human and hamster cells induced them both to express a mutator phenotype and to produce SV40. The mutator could also be activated indirectly by transfecting unirradiated cells with UV damaged calf thymus DNA. In contrast, UV-damaged exogenous DNA failed to rescue SV40 from unirradiated transformed cells. These results suggest that the expression of transforming viruses and of cellular mutator functions is regulated by at least partially independent mechanisms. Unlike the activation of a cellular mutator phenotype, the rescue of SV40 from virus-transformed mammalian cells by UV light might require that the integrated viral DNA and/or specific cellular sequences are directly damaged. PMID- 2991762 TI - Autoantibody to the gastrin receptor in pernicious anemia. AB - We examined serum IgG fractions from 20 patients with pernicious anemia and 25 control subjects for their capacity to inhibit binding of [125I]15-leu human gastrin-17 to parietal-cell-enriched gastric mucosal cells. IgG fractions from six patients reduced gastrin binding by 45.6 +/- 12.2 per cent, as compared with a reduction of 1.8 +/- 0.7 per cent by fractions from the 25 controls. The fractions from these six patients also reduced gastrin-stimulated [14C]aminopyrine uptake by gastric cells (an index of gastric acid secretory activity in vitro) by 50.2 +/- 8.4 per cent (mean +/- S.D.), as compared with 9.2 +/- 4.1 per cent for the controls. IgG fractions from six other patients that did not reduce gastrin binding also inhibited gastrin-stimulated [14C]aminopyrine uptake, by 48.1 +/- 9.1 per cent. These reductions in gastrin binding and aminopyrine uptake were abolished by absorption of the IgG fractions with suspensions of viable gastric mucosal cells but not by absorption with liver or kidney cells. The IgG fractions did not inhibit [3H]histamine binding or histamine-stimulated [14C]aminopyrine uptake. These results suggest that serum IgG from some patients with pernicious anemia contains autoantibodies to the gastrin receptor. PMID- 2991763 TI - Etoposide. PMID- 2991764 TI - Sequential inferior petrosal venous sampling for Cushing's disease. PMID- 2991765 TI - Similarity of vaccinia 28K, v-erb-B and EGF receptors. PMID- 2991767 TI - Alterations of the hprt gene in human in vivo-derived 6-thioguanine-resistant T lymphocytes. AB - Investigations into the extent and significance of somatic gene mutations occurring in vivo in humans have been hampered by the lack of a means of unambiguously defining the mutational origin of in vivo-derived variant cells. Several years ago we proposed that 6-thioguanine-resistant T lymphocytes, present at low frequencies in human peripheral blood, might be useful markers of in vivo somatic mutation. We and others have since described methods for the isolation and study of these unusual cells. The thioguanine-resistant T cell stably lack hypoxanthine-guanine phosphoribosyltransferase (HPRT) activity, suggesting that they are somatic equivalents in normal individuals to cells from individuals with the X-chromosomal hprt Lesch-Nyhan germinal mutation. We now report that in vivo derived thioguanine-resistant T-cell colonies from a single normal individual show a variety of hprt structural alterations, as determined by Southern blot analysis. This finding demonstrates unequivocally that these cells are genetic mutants and validates their use for fundamental and applied mutational studies in humans. PMID- 2991766 TI - Loss of heterozygosity in three embryonal tumours suggests a common pathogenetic mechanism. AB - Children with the Beckwith-Wiedemann syndrome have a greatly increased potential for the specific development of the embryonal tumours hepatoblastoma, rhabdomyosarcoma and Wilms' tumour. Data obtained with molecular probes suggest that the association between these disparate, rare tumour types reflects a common pathogenetic mechanism that entails the somatic development of homozygosity for a mutant allele at a locus on human chromosome 11. PMID- 2991769 TI - More US funds for AIDS. PMID- 2991768 TI - A hybrid recognition sequence in a recombinant restriction enzyme and the evolution of DNA sequence specificity. AB - Early attempts to generate new restriction specificities by recombination between allelic restriction-modification systems have been unsuccessful. Bullas et al. succeeded in isolating a new specificity, SQ, in Salmonella that they interpreted as being the result of a recombination event between the parental strains, Salmonella typhimurium and S. postdam, which encode the SB and SP restriction systems, respectively. This interpretation has recently been confirmed by DNA heteroduplex studies with the SB, SP and SQ structural genes. We have determined the DNA sequences recognized by the SB and SP enzymes and found that, like all type I restriction sequences, they are split into two specific domains by a spacer of nonspecific sequence that, for both SB and SP, is 6 base pairs (bp) long. We have now determined the sequence recognized by the recombinant SQ enzyme and find that it is a hybrid between the SB and SP sequences, containing one specific domain from each parental strain. This result implies that each of the two specific domains is recognized by a physically distinct part of the enzyme. PMID- 2991770 TI - Eukaryotic topoisomerases come into the limelight. PMID- 2991771 TI - SV40 enhancer and large-T antigen are instrumental in development of choroid plexus tumours in transgenic mice. AB - We have shown recently that choroid plexus tumours frequently develop in transgenic mice which have developed from fertilized eggs injected with DNA molecules containing both simian virus 40 (SV40) early-region genes and metallothionein (MT) fusion genes, and several lines of mice have now been established in which all of the offspring that inherit the foreign DNA succumb to these tumours at 3-5 months of age (ref. 1 and our unpublished data). Several other tissues, notably thymus and kidney, occasionally also show pathological changes. SV40 large-T antigen protein and messenger RNA are always present in affected tissues at much greater concentrations than in unaffected tissues, suggesting that SV40 early-region genes are preferentially activated in choroid plexus, thymus and kidney and that this activation frequently leads to tumorigenesis in the choroid plexus. To determine which regions of the original constructs are important for this tumorigenesis, we have now tested several derivatives and report here that the large-T antigen is sufficient, that the MT fusion gene is dispensable and that the SV40 enhancer (72-base-pair repeat region) has an important role in directing tumours to the choroid plexus. Deletion of the SV40 enhancer region alone commonly leads to peripheral neuropathy, as well as liver and pancreatic tumours, which are the subject of the accompanying paper. Evidence is presented that these pathologies may result from an enhancing effect of the MT sequences on large-T antigen genes, made possible by removal of the otherwise dominant SV40 enhancer. PMID- 2991772 TI - Peripheral neuropathies, hepatocellular carcinomas and islet cell adenomas in transgenic mice. AB - The ability to introduce foreign DNA into the genome of mice offers unique opportunities to produce new models of disease process. Recent experiments have shown that integration and expression of simian virus 40 (SV40) T antigen genes and the murine mammary tumour virus (MMTV)-myc genes in transgenic mice can lead to the development of neoplasia in a remarkably tissue-specific manner. In the case of SV40-bearing mice, tumours consistently develop in the choroid plexus. In the accompanying paper, we show that the 72-base pair (bp) enhancer in the SV40 genome is instrumental in directing tumorigenesis to the choroid plexus. However, when the enhancer is deleted from a construction also containing the metallothionein-human growth hormone fusion gene (SV delta e-MGH), an entirely new pattern of pathology results. The present report focuses on transgenic mice carrying this construct; they develop demyelinating peripheral neuropathies, hepatocellular carcinomas and islet cell adenomas. PMID- 2991773 TI - Two tandemly organized human genes encoding the T-cell gamma constant-region sequences show multiple rearrangement in different T-cell types. AB - The recent detailed analysis of genes that undergo rearrangement in T cells has shown that the T-cell receptor genes encoding alpha- and beta-chains are involved in specific alterations in T-cell DNA analogous to the immunoglobulin genes. A third type of gene, designated gamma, has been isolated from mouse cytotoxic T lymphocytes, and evidence suggest that the mouse displays very limited diversity in this gene system, having only three variable-region (V) genes and three constant-region (C) genes. The function of the so-called T-cell gamma gene is unknown. We have isolated genomic genes encoding the human homologue of the mouse T-cell gamma gene; as there is no evidence that this T-cell rearranging gene is anything to do with the T3 molecule, we have designated the human T-cell rearranging gene as TRG gamma (ref. 13), to avoid confusion with the T3 gamma chain, and have shown that the gene locus maps to chromosome 7 in humans. We now report that human DNA contains two tandemly arranged TRG gamma constant-region genes about 16 kilobases apart. These two genes show multiple rearrangement patterns in a variety of T cells, including helper and cytotoxic/suppressor type, as well as in all forms of T-cell leukaemia. Our results indicate variability of this T-cell gene system in man compared with the analogous system in mouse. PMID- 2991774 TI - On the uptake and genotoxicity of UICC Rhodesian chrysotile A in human primary lung fibroblasts. PMID- 2991775 TI - Endogenous noradrenaline as modulator of hippocampal serotonin (5-HT)-release. Dual effects of yohimbine, rauwolscine and corynanthine as alpha-adrenoceptor antagonists and 5-HT-receptor agonists. AB - The modulation of hippocampal serotonin (5-HT)-release by noradrenaline was studied in rabbit hippocampal slices, which were preincubated with 3H-serotonin and then superfused continuously. Electrical field stimulation of the slices elicited a tritium overflow, which was decreased by clonidine in a concentration dependent manner. Phentolamine antagonized the effects of clonidine and, given alone, increased the evoked tritium overflow. This facilitatory effect of phentolamine was further enhanced in the presence of (+)oxaprotilin, a highly selective noradrenaline uptake inhibitor, whereas the (-)enantiomer of oxaprotilin, which does not affect noradrenaline uptake, was inactive. (+)Oxaprotilin but not (-)oxaprotilin, given alone, inhibited the evoked tritium overflow. The inhibitory effect of (+)oxaprotilin was antagonized by phentolamine. In the presence of phentolamine, the alpha-adrenoceptor antagonists yohimbine, rauwolscine and corynanthine decreased the evoked 5-HT-release concentration dependently. Their inhibitory effects were, however, abolished (corynanthine) or inversed to a facilitation of release (yohimbine, rauwolscine) if instead of phentolamine the 5-HT-receptor antagonist metitepin was present. Therefore we suggest that yohimbine, rauwolscine and corynanthine, in addition to their alpha-adrenoceptor antagonistic properties, may act as agonists at 5-HT autoreceptors. Possibly the indol part of their molecules is responsible for this effect. Furthermore, our results provide evidence for the modulation of hippocampal 5-HT-release by endogenous noradrenaline. PMID- 2991777 TI - Effects of isoprenaline and glucagon on insulin secretion from pancreatic islets. AB - The effects of isoprenaline and glucagon on insulin secretion from pancreatic islets were investigated. In the presence of high concentrations of isoprenaline (10-50 mumol/l), glucose-induced (20 mmol/l) insulin secretion from isolated perifused mouse islets was inhibited. This inhibition was apparently mediated by alpha 2-adrenoceptors, as it was antagonized by rauwolscine. At low concentrations isoprenaline (0.1 or 1 mumol/l) did not affect glucose-induced (2.5; 10 or 20 mmol/l) insulin secretion from perifused mouse or rat islets, even if alpha 2-adrenoceptors were blocked by rauwolscine. A stimulatory effect of isoprenaline on insulin secretion was also not observed in the perfused rat pancreas. However, when incubated mouse islets were exposed to glucose (10 mmol/l), insulin secretion was further enhanced by isoprenaline (0.5 mumol/l). To elucidate the underlying mechanism, the effects of glucagon on insulin secretion were investigated, because glucagon is released from the pancreatic A-cells during stimulation with isoprenaline and is accumulated in the islets and the surrounding medium during incubations of pancreatic islets. Indeed, glucagon stimulated insulin secretion from perifused mouse islets in the presence of high glucose (10 or 15 mmol/l) concentrations but not of low glucose (5 mmol/l) concentrations. Thus it is concluded that direct beta-adrenergic stimulation of pancreatic B-cells does not occur in mouse or rat pancreatic islets. Augmentation of glucose-induced insulin secretion by isoprenaline observed in incubation systems can be explained as a result of stimulation by glucagon, which is released from pancreatic A-cells by isoprenaline. PMID- 2991776 TI - Interactions of the enantiomers of 3-O-methyldobutamine with alpha- and beta adrenoceptors in vitro. AB - The enantiomers of 3-O-methyldobutamine, a metabolite of dobutamine, were evaluated for their alpha- and beta-adrenoceptor mediated effects in vitro in a variety of isolated organs and in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed alpha 1-adrenoceptor agonist activity in isolated guinea pig aorta. However, both enantiomers of 3-O-methyldobutamine were competitive alpha 1-adrenoceptor antagonists, with the (+)-enantiomer being approximately 10-fold more potent than the (-)-enantiomer as assessed either in guinea pig aorta or by displacement of 3H-prazosin binding from alpha 1 adrenoceptors in rat cerebral cortex. The alpha 1-adrenoceptor blocking activity of (+)-3-O-methyldobutamine was relatively potent and corresponded to a pA2 of 7.33 in guinea pig aorta and a -log Ki of 7.72 in radioligand binding studies. Neither enantiomer of 3-O-methyldobutamine possessed alpha 2-adrenoceptor agonist activity in field-stimulated guinea pig ileum. Although (+)-3-O-methyldobutamine weakly inhibited the twitch response in field-stimulated guinea pig ileum, the response was not blocked by the selective alpha 2-adrenoceptor antagonist, yohimbine, and was found to result from weak anticholinergic activity (pA2 = 5.06). Neither enantiomer of 3-O-methyldobutamine possessed beta 1-adrenoceptor agonist activity in guinea pig atria, however the (+)-enantiomer was a weak noncompetitive antagonist at beta 1-adrenoceptors. In contrast, both enantiomers of 3-O-methyldobutamine were weak beta 2-adrenoceptor agonists in rat uterus, however these weak effects were not highly stereoselective, which was also confirmed in radioligand binding studies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991778 TI - The interaction of canrenone with the Na+,K+ pump in human red blood cells. AB - Canrenone inhibits 30-40% of ouabain-sensitive Na+ efflux in human red cells. Half-maximal inhibition was obtained with a canrenone concentration = 86 +/- 37 mumol/l (mean +/- SD of 13 experiments). The partial inhibition of the Na+,K+ pump appears to be mediated at the digitalis receptor site with an apparent dissociation constant (Kc) = 200 +/- 130 mumol/l (mean +/- SD). Further evidence suggesting that canrenone is a partial agonist at the digitalis receptor site was obtained by the observation that it decreases the apparent affinity of the Na+,K+ pump for external K+. However, in contrast to ouabain, canrenone decreases the apparent pump affinity for internal Na+. Our results show that, at physiological cell Na+ levels canrenone is able to enhance the inhibition of the Na+,K+ pump by low doses of ouabain. Conversely, in cells treated with high concentrations of cardiac glycosides (in which cell Na+ content increases), canrenone is able to restimulate the blocked pumps. PMID- 2991780 TI - [New methods, new diseases; benign liver tumors]. PMID- 2991781 TI - [Significance of electron microscopy in the histological classification of lung carcinoma]. PMID- 2991779 TI - DPI 201-106, a novel cardioactive agent. Combination of cAMP-independent positive inotropic, negative chronotropic, action potential prolonging and coronary dilatory properties. AB - The in vitro cardiac effects of DPI 201-106, a novel piperazinyl-indole, were investigated. DPI 201-106 produced concentration-dependent positive inotropic effects in guinea-pig and rat left atria, kitten, rabbit and guinea-pig papillary muscles and Langendorff perfused hearts of rabbits between 10(-7) and 3 X 10(-6) mol/l. During isometric twitches, contraction and relaxation phases were prolonged in guinea-pig left atria and right ventricular papillary muscles from kitten and guinea-pigs. Spontaneous sinus rate was decreased in right atria of guinea-pigs and rats. Coronary flow increased in rabbit isolated hearts. Functional refractory period was increased in left atria from guinea-pigs and rats with EC50 values of 1.7 and 0.24 mumol/l respectively. In electrophysiological measurements, DPI 201-106 prolonged the action potential duration (APD70) in guinea-pig papillary muscles up to 70% and in rabbit atria up to 120% at 3 mumol/l. Other action potential characteristics were not changed in guinea-pig papillary muscles but Vmax was decreased in rabbit left atria. The electrophysiological as well as the positive inotropic effects were stereoselective with the activity residing in the S-enantiomer. DPI 201-106 increased the Ca2+-sensitivity of skinned fibres from porcine trabecula septomarginalis with an EC50 of 0.2 nmol/l. DPI 201-106 dit not change cAMP levels in guinea-pig atria and rabbit papillary muscles. Slow action potentials were not induced by DPI 201-106 in partially depolarized guinea-pig papillary muscles. Phosphodiesterase activity of rat hearts was not inhibited by DPI 201 106 at pharmacologically relevant concentrations. The presence of propranolol did not influence the inotropic potency of DPI 201-106 in guinea-pig atria. In conclusion, DPI 201-106 represents a novel type of positive inotropic agents with a synergistic sarcolemmal and intracellular mechanism of action. PMID- 2991782 TI - [DNA analysis and genetic counseling]. PMID- 2991783 TI - [Search for the carrier state of hemophilia B using restriction fragment length polymorphism]. PMID- 2991785 TI - [A difficult diagnosis?]. PMID- 2991784 TI - [Linitis plastica, a rare disease picture]. PMID- 2991786 TI - [Features of projections of afferent fibers of the ventral roots to neurons of the dorsal horn of the spinal cord in the cat]. AB - Some peculiarities of afferent effects from ventral roots on the dorsal horn neurons were investigated in acute experiments on cats. Convergence of afferents from ventral and dorsal roots on the same neurons was shown. Excitation of some ventral root afferents caused inhibition in investigated neurons. This inhibition developed with shorter latency than that from dorsal root afferents. PMID- 2991787 TI - [Does non-quantum secretion of acetylcholine from motor neuron endings participate in the neurotrophic control of the membrane potential of muscle fibers in the rat?]. AB - The resting membrane potential of synaptic zone of muscle fibres within 2-3 mm (near) and 9-11 mm (far) of the nerve section of the rat diaphragm muscle was measured after 3 hours or on the 5th day after the motor nerve section. The membrane potential of "near" fibres was lower than that of "far" fibres. The presence of carbamylcholine or cGMP in the culture medium maintained the membrane potential of "near" fibres close to that of "far" fibres. Ouabain prevented these effects. On the 5th day after the nerve section the presence of carbamylcholine or cGMP had no effect on the membrane potential. In the in vitro experiments the nonquantal acetycholine release was equal in "near" and "far" fibres and just the same as in the control preparations. A conclusion is made that synaptic acetylcholine from motor nerve endings in nonquantal form does not play a significant role in neurotrophic control of the resting membrane potential of muscle fibres membrane in mammals. PMID- 2991788 TI - [GABA-ergic structures in intact and chronically isolated association cortex of the cat brain (field 5)]. AB - Distribution of GABAergic structures in intact and neuronally isolated associative cortex (field 5) of the cat was studied using histochemical GABA transaminase reaction two and three weeks after isolation. An overwhelming majority of GABAergic fibres of the neuropile and synaptic terminals were found to be formed by axons of few GABAergic interneurons. Only a small portion of them belongs to afferent axons of the extracortical origin. GABAergic interneurons can be divided into short-axon ones, making contacts within the isolated slat, and long-axon neurons forming horizontal connections with more distant cortical neurons. GABAergic axons give numerous projections to the soma and proximal parts of dendrites of many pyramidal neurons, containing no GABA-transaminase, and of stellate neurons which are of GABAergic and non-GABAergic mediatory nature. It is suggested that the influence of some GABAergic neurons on the others may ensure intracortical spatial regulation of inhibitory processes. PMID- 2991789 TI - [Generation of locomotor rhythms in Limacina helicina]. AB - Two groups of neurons (motoneurons and putative interneurons), exhibiting periodic activity with the locomotory rhythm, were recorded in the pedal ganglia of the isolated nervous system of the mollusc Limacina helicina. Motoneurons periodically generated spike bursts, while interneurons generated only one prolonged (100-400 ms) action potential per cycle. Rhythmic generation persisted after blocking the spike discharges of motoneurons by means of tetrodotoxin. Rhythmic generation could be facilitated by application of serotonin. PMID- 2991790 TI - [Posttetanic potentiation at an identified synapse of the central nervous system of Helix lucorum during different types of tetanization]. AB - Posttetanic potentiation (PTP) of the identified synapse (LPa9-LPa3) in the central nervous system of snail (Helix lucorum L.) was investigated. PTP was observed both after tetanization of presynaptic neuron LPa9 by intracellular current pulses and after tetanization of the same presynaptic neuron by extracellular stimulation of its axonal branch. PMID- 2991791 TI - [Brain tumor induced in dogs by intracerebral inoculation of SR-RSV induced cultured tumor cells--electron microscopic study]. AB - An experimental transplantable canine brain tumor model with the advantages of rapid tumor growth within 10 days and relative safety for the investigator is presently available. The tumor is produced by intracerebral inoculation of cultured cells derived from a canine brain tumor induced by the Schmidt-Ruppin strain of the Rous-Sarcoma virus (SR-RSV). It has potential use as a model in experiments designed to evaluate the effectiveness of chemotherapy and radiotherapy with serial computerized tomography scans. However, characterization of the induced tumor is essential. Ideally, it should have features attributable to glioma and/or neuroectodermal tumors. Utilizing the technique of intracerebral inoculation of cells cultured from the original dog brain tumor induced by SR RSV, Salcman et al identified the tumor they induced in brains of mongrel puppies as a glioma by light microscopic criteria (Reference). The purpose of our study was to further characterize this experimental tumor by electron microscopic and immunohistochemical techniques. Tumor was induced in 6 mongrel puppies. Stains of the tumor for immunohistochemical reactivity to glial fibrillary acid protein, S 100 protein and 210K neurofilament protein were all negative. With the electron microscope, the intracerebral tumor cells were mostly undifferentiated. They had a few cell processes, occasional punctate adhesions and some microvilli-like structure. The tumor cell nucleus was usually oval shaped and sometimes had nuclear indentations. The cytoplasm contained abundant free ribosomes, some rough endoplasmic reticulum and mitochondria. Collagen fibers and basal lamina were not observed in the intercellular spaces. The capillaries within the tumor were characterized by proliferation of immature endothelial cells which were non fenestrated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991792 TI - [Extraneural metastasis of malignant glioma through a ventriculo-peritoneal shunt: growth in peritoneal cavity as ascitic form]. AB - This is the first report of extraneural metastasis of malignant glioma through V P shunt tube and growth in peritoneal cavity as ascitic form. The patient was a 43-year-old man who was admitted to our hospital with occipital headache. CT scan showed enhanced cystic tumor mass at left temporal lobe. Craniotomy and partial excision of the tumor was done and the histology of tumor tissue showed a malignant astrocytoma. Following this treatment, the patient received the adjuvant therapies of radiation, chemotherapy and immunotherapy with interferon, and also recraniotomy three times. In the mean time, a ventriculo-peritoneal shunt was set up for internal hydrocephalus. One month later, abdominal bulging appeared and yellowish ascites could be obtained with peritoneal tap. In the ascite, tumor cells with glial fibrillary acidic protein were observed at the concentration of 5-10 x 10(4) cells/ml. The patient died three months after extraneural metastasis to the abdominal cavity as ascitic form. At autopsy, solid metastatic mass lesion was not found in extraneural region include abdomen. PMID- 2991793 TI - Glycogen content and fructose-1, 6-biphosphatase activity in methionine sulfoximine epileptogenic mouse brain and liver after protein synthesis inhibition. AB - Mice given intraperitoneal injections of methionine sulfoximine (MSO) (100 mg/kg body weight) showed tonic-clonic seizures 7 to 8 h later. The protein synthesis inhibitors actinomycin D and cycloheximide, when combined with MSO delayed the onset of seizures. Methionine completely abolished the convulsions and metyrapone delayed them for some hours. Twenty-four h after the administration of the convulsant, the activity of the gluconeogenic enzyme, fructose-1, 6-biphosphatase (FBPase), and the glycogen content were determined in different areas of the brain. MSO induced an increase in both FBPase activity and glycogen content. These effects were antagonized by the inhibitors of protein synthesis. Metyrapone partly inhibited MSO-induced increases of FBPase activity and glycogen content whereas methionine completely abolished them. MSO decreased glycogen content in liver but had no effect on blood glucose level 24 h after its administration. These findings suggested that in MSO epileptogenic brain, glycogen accumulation may proceed from an enhanced gluconeogenesis. PMID- 2991794 TI - Evidence for the modulation of sexual behavior by alpha-adrenoceptors in male rats. AB - Clonidine, a commonly used antihypertensive agent believed to act by stimulation of central alpha-adrenoceptors, produced a dose-related suppression of ejaculatory behavior in sexually vigorous male rats throughout the range of treatment (0.0125-0.5 mg/kg). Treatment with 0.25 mg/kg virtually eliminated ejaculatory behavior, without altering the number of animals mounting and intromitting. These effects were maintained for at least 4 h after treatment. Prazosin, another antihypertensive (but acting by blockade of alpha 1 adrenoceptors), increased latencies to initiation of copulation, to ejaculation, and to reinstatement of copulation following ejaculation. Additionally, prazosin (1 mg/kg) pretreatment failed to attenuate or prevent the clonidine-induced suppression of ejaculation. In contrast, yohimbine, a drug which preferentially blocks alpha 2-adrenoceptors (2 mg/kg, 20 min prior to mating tests), caused a facilitation of copulatory behavior as evidenced by drastic decreases in ejaculation latency and intercopulatory and postejaculatory intervals. Pretreatment with yohimbine completely prevented the clonidine-induced suppression of ejaculation, while clonidine attenuated the facilitatory effects of yohimbine, suggesting a competitive interaction. These data lead to the suggestion that increased excitatory adrenergic activity results in increased sexual arousal either by blockade of alpha 2-or stimulation of alpha 1 adrenoceptors. Alternatively, stimulation of alpha 2-or blockade of alpha 1 adrenoceptors results in diminished sexual motivation. While the precise implications of this animal research for the clinic are as yet unclear, further research could lead to important developments in the pharmacological treatment of sexual dysfunctions. PMID- 2991795 TI - Castration affects male rat brain opiate receptor content. AB - We previously reported that saturable stereospecific binding of [3H]-naltrexone in rat brain homogenates prepared from castrated male rats was greater than the corresponding binding in intact animals. We now report that we have replicated these results and that the difficulty of other investigators in observing these differences is due to methodological factors. Specifically, when samples were filtered individually and rapidly, differences between castrated and intact rats were maintained. The increase in binding was also observed when tissues were washed to remove endogenous opioids prior to incubation, when [3H]-naloxone was used as the ligand, and when various antagonists were used as displacers in the radioreceptor assay. PMID- 2991796 TI - Streptozotocin-induced diabetes is associated with reduced immunoreactive beta endorphin concentrations in neurointermediate pituitary lobe and with disrupted circadian periodicity of plasma corticosterone levels. AB - Lower concentrations of immunoreactive (IR) beta-endorphin were present in the neurointermediate pituitary lobes of streptozocin-induced diabetic versus control animals at both 2 and 4 weeks after the onset of diabetes. The forms of beta endorphin-like material present appeared to be similar in both groups when studied with cation-exchange chromatography. Insulin therapy via minipump for 2 weeks did not alter this finding of lowered beta-endorphin concentrations in diabetic animals, despite normalization of blood glucose levels and body weight gain. Lower IR beta-endorphin levels were also found in neurointermediate lobes of weight-restricted rats, but this group had increased plasma IR beta-endorphin concentrations compared to diabetic animals. Concentrations of IR beta-endorphin in microdissected brain regions and in anterior pituitaries of the diabetic animals failed to show consistent changes; in addition, ACTH concentrations in pituitary lobes and plasma did not differ among groups. Circadian rhythmicity of plasma insulin and corticosterone concentrations was absent in the diabetic animals, although food and water intake, while elevated, showed the normal nocturnal pattern of increased ingestion. Furthermore, adrenal hypertrophy was present in the diabetic animals and was accompanied by an elevation of mean plasma corticosterone levels. The present findings indicate that diabetes is associated with a decrease of neurointermediate pituitary lobe synthesis of beta endorphin, while not affecting the processing of the peptide in this lobe, and confirm previous reports of altered adrenal function in diabetic animals. PMID- 2991797 TI - Calmodulin inhibitors decrease the CRF-and AVP-induced ACTH release in vitro: interaction of calcium-calmodulin and the cyclic AMP system. AB - The effect of N-(6-aminohexyl)-5-chloro-naphthalene-1-sulfomide (W-7) and trifluoperazine (TFP) was examined on ACTH release from cultured rat anterior pituitary cells and pituitary halves. These drugs significantly inhibited the ACTH release induced by synthetic ovine corticotropin-releasing factor (CRF) in a dose-related manner. In pituitary halves, arginine vasopressin (AVP) at 10 and 100 ng/ml showed almost the same ACTH-releasing activity as CRF at the same concentrations. W-7 and TFP inhibited the CRF-and AVP-induced ACTH release from pituitary halves. The cyclic AMP levels in the pituitary halves were significantly increased by CRF, but not AVP. Although W-7 inhibited CRF-induced ACTH release, it did not have an effect in cyclic AMP accumulation. These results suggest that CRF exerts ACTH-releasing activity through both the calcium calmodulin system and cyclic AMP system and that AVP stimulates ACTH release mainly through the calcium-calmodulin system. PMID- 2991798 TI - Effect of acute ether stress on monoamine metabolism in median eminence and discrete hypothalamic nuclei of the rat brain and on anterior pituitary hormone secretion. AB - This study was designed to correlate the endocrine responses elicited by acute ether stress with the changes in metabolism of several monoamines in discrete nuclei of the rat brain. Concentrations of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT) and also of the specific metabolites of NE, DA, and 5-HT, 3-methoxy-4-hydroxyphenylethylene glycol, 3,4-dihydroxyphenylacetic acid, and 5-hydroxyindole-3-acetic acid, respectively, were concurrently measured in microdissected nuclei using high-performance liquid chromatography with electrochemical detection. The ratio of the metabolites to their respective amines was used as an estimate of the metabolism of NE, DA, and 5-HT. Acute exposure to ether vapors induced, within 5-15 min, large increments in plasma levels of adrenocorticotropic hormone (ACTH), beta-endorphin, and prolactin (PRL), and decrements in the levels of plasma growth hormone (GH). Significant increases in NE metabolism were observed in the rostral (ANr) and caudal (ANc) divisions of the arcuate nucleus, as well as in the paraventricular (PVN) and dorsomedial nuclei, 15 min after ether stress. A significant decrease in 5-HT metabolism was observed in the PVN, supraoptic nucleus, and ANc, whereas significant increases in 5-HT metabolism were detected in the suprachiasmatic nucleus and ANr. DA metabolism selectively increased in the ANr. The present results indicate that the acute changes in ACTH, beta-endorphin, PRL, and GH release induced by ether exposure are temporally correlated with increases in NE metabolism in many hypothalamic nuclei; a selective increase in DA metabolism restricted to the ANr, and differential effects on 5-HT metabolism, probably reflecting selective activation or inhibition of different populations of 5-HT neurons. PMID- 2991799 TI - An analysis of the effects of clonidine and three structural isomers on transmission in rat superior cervical ganglia in vitro. AB - alpha-Adrenoceptors mediating inhibition of release of acetylcholine in the superior cervical ganglia of the rat have previously been characterized as of the alpha 2-subtype (Brown and Caulfield, 1981). In the present study, the effects of clonidine (2,6-dichlorophenylimino-2-imidazolidine), the 2,3- 2,4- and 2,5 dichloro isomers (St 476, St 363 and St 475) and the 3,4-dihydroxy analogue (DPI) were examined on transmission in this tissue. Compound action potentials (CAP; supramaximal preganglionic stimulation at 0.2 Hz) were recorded extracellularly. Clonidine, DPI and adrenaline inhibited the compound action potential; concentration-response curves were shifted to the right by phentolamine. The maximum inhibition of the compound action potential was similar for epinephrine and DPI. However, for clonidine a secondary inhibitory component, leading to almost complete blockade of the compound action potential, was seen in concentrations greater than 10 microM; similar effects were seen for the lipid soluble analogues St 476, St 363 and St 475, suggesting a non-specific effect. In smaller concentrations, St 476, St 363 and St 475 exerted only weak inhibition of the CAP; St 363 and St 475 actually increased the height of the compound action potential, in a similar manner to phentolamine (0.1-3 microM), and all three analogues blocked the inhibitory effects of adrenaline. These latter data suggest partial agonist actions for St 476, St 363 and St 475. The rank order of inhibitory effectiveness (clonidine greater than St 476 greater than St 363 greater than St 475) parallels that for acute hypotensive effects in rats (Timmermans and van Zwieten, 1977a). The results suggest that sympatho-inhibitory effects of some imidazolidines may not result solely from activation of presynaptic alpha 2-adrenoceptors. PMID- 2991801 TI - Evidence for GABA tolerance in barbiturate-dependent and withdrawn mice. AB - Mice were rendered tolerant and dependent to barbital by a chronic feeding schedule of barbital over 5 weeks. The behavioural effects of muscimol, imidazole acetic acid (ImAA), and gamma-hydroxybutyric acid (GHB) were measured in control, barbital-dependent, and mice dependent on barbital 48 hr after withdrawal of the drug. The sedative effects of the GABA-mimetics imidazole-acetic acid and muscimol were increased in dependent mice, but reduced in withdrawn mice. Subanaesthetic doses of barbital, given acutely, also increased the sedative effects of imidazole-acetic acid and muscimol but not of gamma-hydroxybutyric acid. Assay of plasma barbital levels by GLC indicated that a negligible amount of barbital was present 48 hr after withdrawal compared to levels of between 60 and 120 micrograms/ml during chronic treatment with barbital. The binding of [3H]GABA to membrane preparations from brain indicated that the values of Kd and Bmax for low affinity binding were not significantly altered in mice withdrawn from chronic treatment with barbital, but that the Kd for high affinity binding was significantly increased from 4.38 to 6.06 nM in barbital-withdrawn mice. There was no difference in the enhancement of GABA binding by pentobarbital between the two groups. It is concluded that barbital-tolerant and dependent mice are cross-tolerant to GABA and that this is possibly mediated by a change in the affinity of the GABA receptor for its ligand. PMID- 2991802 TI - Adenosine: an endogenous modulator of hippocampal noradrenaline release. AB - In slices of hippocampus from the rabbit, preincubated with [3H]noradrenaline and then continuously superfused, the modulation of the release of noradrenaline by adenosine receptors was studied. Electrical field stimulation of the slices elicited a release of [3H]noradrenaline which was inhibited in a concentration dependent manner by various adenosine receptor agonists. From the order of potency: cyclohexyladenosine greater than (-)phenylisopropyladenosine [(-)PIA] greater than 5'-N-ethylcarboxamide-adenosine (NECA) greater than 2-chloro adenosine greater than adenosine (+)phenylisopropyladenosine greater than ATP, the inhibitory adenosine receptor was classified as A1- (Ri-) receptor. The effect of the agonist was strongly reduced by adenosine receptor antagonists, the methylxanthines. A role for endogenous adenosine in the modulation of hippocampal noradrenaline release is supported by these findings: (1) that blockade of adenosine receptors by methylxanthines, especially by 8-phenyltheophylline, increased, whereas (2) inhibition of the uptake of adenosine decreased the evoked release of noradrenaline and (3) that deamination of endogenous extracellular adenosine by addition of adenosine deaminase to the medium enhanced the evoked transmitter release. Inhibitors of endogenous adenosine deaminase and 5' nucleotidase were without effect. It is concluded that release of noradrenaline in the hippocampus is inhibited at the level of the noradrenergic nerve terminals by endogenous adenosine via A1 (or Ri) receptors. PMID- 2991800 TI - Oxytocin and a C-terminal derivative (Z-prolyl-D-leucine) attenuate tolerance to and dependence on morphine and interact with dopaminergic neurotransmission in the mouse brain. AB - The effects of oxytocin (OXT) and of dipeptides derived from the C-terminal portion of oxytocin (Z-prolyl-leucine and Z-prolyl-D-leucine) on the development of acute and chronic tolerance to, and dependence on morphine were tested in the mouse. Oxytocin and the dipeptides attenuated the development of acute and chronic tolerance to the antinociceptive effect of morphine and delayed the onset of the naloxone-precipitated withdrawal syndrome. Both oxytocin and Z-prolyl-D leucine affected drug-induced behavioural responses related to dopamine (DA) in the brain. Thus, oxytocin potentiated the hypermotility induced by a large dose of apomorphine and decreased the supersensitivity of the DA receptors. Small doses of Z-prolyl-D-leucine inhibited the hypomotility elicited by a small dose of apomorphine and potentiated the hyperactivity induced by amphetamine. The data indicate that both oxytocin and Z-prolyl-D-leucine affect tolerance to and dependence on morphine. While oxytocin interacts mainly with postsynaptic DA ergic neuronal elements, the dipeptide primarily affects DA-ergic neurotransmission at the presynaptic level. PMID- 2991803 TI - Classification of ischemic-induced damage to Na+, K+-ATPase in gerbil forebrain. Modification by therapeutic agents. AB - The activity of three forms of ATPase were examined in fractions of the brain of the gerbil treated with ethylene glycol-N-N-tetra-acetic acid (EGTA) under a variety of conditions of primary and secondary (reflow) ischemia. In animals which were unilateral ischemic (ligation of the right common carotid), damage to Na+, K+-ATPase alone was observed only after at least 6 hr of ischemia had elapsed. The phenomenon occurred in only symptomatic gerbils and was absent in animals which were either asymptomatic or only displayed partial neurological symptoms. Under conditions of bilateral cerebral ischemia, in which both carotid arteries were clamped, only irreversible ischemia (60 min) followed by reflow, was associated with highly significant damage to cerebral Na+, K+-ATPase. In regional studies of the forebrain involving ischemia for 60 min plus 30 min reflow, damage to Na+, K+-ATPase was evident in the cerebrum, hippocampus, striatum and thalamus, while the hypothalamus and olfactory bulb were spared. Pretreatment of gerbils with allopurinol, clonazepam or combinations of thiopental plus either indomethacin or methylprednisolone offered protection to cerebral Na+, K+-ATPase subsequent to secondary ischemia. With only minor exceptions (striatum) neither Ca2+, Mg2+- nor Mn2+-ATPase were altered by stroke or treatment with drugs. PMID- 2991805 TI - Verapamil alters the amplitude and time course of miniature endplate current. AB - The effect of verapamil on neuromuscular transmission was examined by recording miniature endplate currents (mepcs) in voltage-clamped frog sartorius muscle fibres. In the presence of 100 microM verapamil, the amplitude and time constant of decay of the mepcs (tau D) were reduced to 68% and 55% of control respectively, and the normal voltage dependency of tau D was decreased. Part of the decrease in amplitude of the mepc was independent of changes in tau D because, on washout, recovery of tau D was more rapid than that of amplitude, and in some cells smaller concentrations of verapamil (1 and 10 microM) decreased amplitude without affecting tau D. Evidence of open and closed channel blockade by verapamil (5-20 microM) was obtained from ionophoretic end-plate current trains and it is proposed that these effects are mediated via an allosteric mechanism. PMID- 2991804 TI - Acetylcholine hydrolysis during neuromuscular transmission in the synaptic cleft of skeletal muscle of mouse and chick. AB - Inhibition of acetylcholinesterase (AChE) by more than 80% by neostigmine or physostigmine resulted in a failure of tetanic contraction (100 Hz) in the isolated mouse nerve-diaphragm preparation. In the chick biventer cervicis muscle, however, the tetanic contraction was well maintained and even outlasted the period of nerve stimulation after inactivation of AChE. The concentration of (+)tubocurarine for 70% block of the indirect twitch response of the mouse diaphragm at 0.1 Hz was increased from 0.67 to 0.99 to 1.21-2.03 microM in the presence of neostigmine (0.15-1.5 microM) which inhibited AChE by 70% or more, while that to depress the tetanic contraction (50 Hz) was increased from 0.38 to 0.42 to 0.53-0.69 microM. In the chick muscle, physostigmine at 2.4 microM increased the concentration of (+)tubocurarine for 70% block of the twitch response from 1.68 to 4.14 microM, whereas that for block of the response to exogenous acetylcholine (ACh) was increased from 1.47 to 74.6 microM. On single stimulation, the relative peak concentrations of acetylcholine (ACh) at the postsynaptic receptor site of the mouse diaphragm and chick biventer cervicis were estimated to be increased by about 110 and 120% respectively, after complete inhibition of AChE. In the chick muscle, physostigmine increased the relative concentration of ACh by about 40-fold at the receptor site for exogenously applied ACh. It is concluded that the intrinsic ACh released from the nerve terminal is hydrolyzed by about 50% during the time of diffusion across the synaptic cleft whereas most of exogenous ACh is hydrolyzed before reaching the target. PMID- 2991806 TI - The role of central dopaminergic systems in the behavioral effects of H-Phe-Ile Tyr-His-Ser-Tyr-Lys-OH. AB - In the present study the role of the central dopaminergic systems in the behavioral action of H-Phe-Ile-Tyr-His-Ser-Tyr-Lys-OH was investigated. The heptapeptide inhibited the extinction of active avoidance behavior if the treatment was performed intracerebroventricularly (icv) in a dose of 1 microgram, but was ineffective in a dose of 0.1 micrograms. If the peptide was injected into the nucleus accumbens septi (NAS) in a dose of 1 microgram, 0.1 micrograms or 0.01 micrograms it inhibited the extinction, but in the latter dose its effect was only a short one. H-Phe-Ile-Tyr-His-Ser-Tyr-Lys-OH in a dose of 1 microgram, 10 micrograms or 20 micrograms (icv) did not influence the turning activity of unilateral substantia nigra (USN)-lesioned animals. These results suggest that the NAS plays an important role in the behavioral action of H-Phe-Ile-Tyr-His-Ser Tyr-Lys-OH and the heptapeptide has no direct dopamine receptor stimulatory or dopamine-releasing effect in the striatum. PMID- 2991807 TI - Evidence that the delta-selective alkylating agent, fit, alters the mu noncompetitive opiate delta binding site. AB - Considerable evidence supports the notion that the prototypic delta agonist [3H]D ala2-D-leu5-enkephalin labels two binding sites on brain membranes in vitro. Recent studies have demonstrated that treatment of brain membranes with the delta selective, site-directed, alkylating agent, FIT (Rice et al., Science 220, 314 316, 1983) results in a membrane preparation devoid of detectable higher affinity [3H]D-ala2-D-leu5-enkephalin binding sites, but contain residual lower affinity binding sites at which mu-ligands are apparent noncompetitive inhibitors (Rothman et al., Neuropeptides 4:210-215, 1984). In this paper we extend these data by showing that although FIT eliminates the higher affinity binding site, it also alters the properties of the residual lower affinity binding sites. PMID- 2991808 TI - Cystic supratentorial gliomas: natural history and evaluation of modes of surgical therapy. AB - The management of cystic supratentorial gliomas is hampered by lack of documentation of the natural history of these lesions and by a lack of evaluation of modes of surgical therapy. We analyzed these factors in 25 patients with solitary cysts operated upon over a 20-year period. Two distinctive patterns of symptoms were seen: short duration (increased pressure and hemiparesis), most often heralding a malignant lesion, and long duration (commonly seizure disorder), associated more often with a benign pathological condition. Large solitary cysts were found in tumors of all histological grades. Surgical procedures included extirpation, biopsy/partial resection, cyst communication to ventricle or marsupialization, burr hole aspiration, aspiration via an indwelling reservoir, and cyst-peritoneal shunting. Radiotherapy, given in all cases, did not prevent cyst recurrence. Of the 25 patients, 76% are alive and remain cyst free at follow-up intervals of 1 to 16 years (mean, 3.2). Five patients died from their tumors, with a mean survival of 33 months after decompression. In 7 of 8 patients with cysts largely or entirely within the basal ganglia or thalamus, successful operative cyst control was achieved. Patients with solitary cystic gliomas seem to have a favorable prognosis, and vigorous efforts to control cyst recurrence and limit disability are warranted. Analysis of our data suggests that craniotomy for tumor resection, cyst decompression, and tissue diagnosis is the initial procedure of choice. Cyst recurrence without major solid tumor should be controlled by computed tomography-guided tap or shunt drainage. Reexploration is indicated when cyst reaccumulation is accompanied by clear regrowth of a solid component. PMID- 2991809 TI - Use of nicotinamide and pyridoxal-5-phosphate in the treatment of experimental epilepsy. PMID- 2991810 TI - Possibilities and limitations of the H-reflex test in the diagnosis of diseases of the nervous system. PMID- 2991811 TI - Reduction of inhibition by a benzodiazepine antagonist, Ro15-1788, in the rat hippocampal slice. AB - The effects of extracellular applications of benzodiazepine agonists and the benzodiazepine antagonist, Ro15-1788, were investigated on pyramidal neurons in the CA1 region of rat hippocampal slices. The benzodiazepine agonists, chlordiazepoxide and diazepam, enhanced gamma-aminobutyrate synaptic inhibition, as tested by extracellular recordings during a paired-pulse inhibition paradigm. In contrast, Ro15-1788 (0.1-1 microM) depressed paired-pulse inhibition in a dose dependent manner that suggested agonist activity at higher (10-100 microM) concentrations. Intracellular recordings from CA1 neurons showed that Ro15-1788 reduced both orthodromically and antidromically evoked inhibitory postsynaptic potentials. The reduction of the inhibitory postsynaptic potential probably resulted from a postsynaptic effect on the conductance mechanism of the inhibitory postsynaptic potential, since there were no changes in resting input resistance, the inhibitory postsynaptic reversal potential or the frequency of spontaneous inhibitory postsynaptic potentials. These data suggest that in the hippocampal slice preparation either (1) an endogenous benzodiazepine agonist exists that can be displaced by Ro15-1788 or (2) Ro15-1788 has inverse agonist activity. PMID- 2991812 TI - The localization of adrenoceptors and opiate receptors in regions of the cat central nervous system involved in cardiovascular control. AB - The distribution of adrenoceptors and opiate receptors in the nucleus of the tractus solitarius and the intermediolateral cell column of the thoracic spinal cord of the cat have been investigated using an in vitro autoradiographic technique. Specific binding of [3H]yohimbine and [3H]rauwolscine (alpha 2 adrenoceptor ligands) was seen within the intermediolateral cell column but no obvious binding of [3H]prazosin, an alpha 1-ligand, was observed. No evidence of a significant population of opiate receptors was obtained in the intermediolateral cell column. Within the nucleus of the tractus solitarius a marked binding of [3H]yohimbine and [3H]rauwolscine was accompanied, however, by a more restricted binding of [3H]naloxone and [3H]dihydromorphine indicating the presence of both alpha 2-adrenoceptors and opiate receptors. As with the intermediolateral cell column no evidence of [3H]prazosin binding was seen. These observations may have particular relevance for the physiology and pharmacology of cardiovascular control. In the case of the intermediolateral cell column it is consistent with evidence of a catecholamine innervation originating from the brainstem. With regard to the nucleus of the tractus solitarius the location of the receptor groups is discussed in the light of the anatomy and physiology of its afferent innervation. PMID- 2991813 TI - Peripheral neuropathy in hypereosinophilic syndrome. AB - We evaluated seven patients with the hypereosinophilic syndrome (HES) to define the clinicopathologic spectrum of the peripheral neuropathy. Clinically, three had evident polyneuropathy; the others were asymptomatic, although they had electrophysiologic evidence of neuropathy. Nerve conduction studies and EMG were compatible with axonal neuropathy. Morphometry of sural nerves from four patients ranged from normal to marked axonal loss, more prominent in large myelinated fibers. Demyelination was rare, and there was no evidence of vasculitis. Neuropathy may be produced by an eosinophil-derived neurotoxin. PMID- 2991814 TI - Tropical myeloneuropathies: the hidden endemias. AB - Tropical myeloneuropathies include tropical ataxic neuropathy and tropical spastic paraparesis. These disorders occur in geographic isolates in several developing countries and are associated with malnutrition, cyanide intoxication from cassava consumption, tropical malabsorption (TM), vegetarian diets, and lathyrism. TM-malnutrition was a probable cause of myeloneuropathies among Far East prisoners of war in World War II. Clusters of unknown etiology occur in India, Africa, the Seychelles, several Caribbean islands, Jamaica, and Colombia. Treponemal infection (yaws) could be an etiologic factor in the last two. Tropical myeloneuropathies, a serious health problem, are multifactorial conditions that provide unsurpassed opportunities for international cooperation and neurologic research. PMID- 2991815 TI - Angiotensin converting enzyme in cerebrospinal fluid: a new assay. AB - Serum and CSF angiotensin converting enzyme (ACE) were measured by a new inhibitor binding assay in 32 patients with sarcoidosis, 49 with neurologic diseases, and 38 controls. In neurosarcoidosis, 11 of 20 patients had high levels of CSF ACE. In systemic sarcoidosis without neurologic abnormality, only 1 of 12 patients had elevated CSF ACE. The highest value was observed in a patient with widespread meningeal sarcoidosis. High values were also observed in patients with bacterial meningitis or malignant tumors of the CNS. Fluctuation in successive analyses correlated to clinical course of neurosarcoidosis. CSF ACE analysis seems useful in diagnosis and follow-up of neurosarcoidosis. PMID- 2991816 TI - Peripheral neuropathy in abetalipoproteinemia. AB - We studied the peripheral neuropathy of three sisters with abetalipoproteinemia. Clinically, a sensory neuropathy progressively increased in severity. There was a diminution in the amplitude of sensory action potentials and a slight-to-moderate slowing in maximum sensory conduction velocity, initially most marked in distal portions of the nerves. Motor conduction was normal, although EMG indicated subclinical signs of partial chronic denervation. The sural nerves showed a decreased number of large fibers (greater than 7 micron); in the patient with the neuropathy of shortest duration, small fibers and clusters of regenerating fibers indicated regeneration. In the two patients with advanced neuropathy, one-half the segments of teased fibers showed paranodal demyelination. Also, unmyelinated fibers showed evidence of regeneration. PMID- 2991817 TI - Treatment of Alzheimer's disease with a synthetic ACTH 4-9 analog. AB - The synthetic ACTH 4-9 analog, Org 2766, was administered to 77 Alzheimer's patients for 6 months in a double-blind, placebo-controlled study. Although Org 2766 was tolerated well, no significant effect of Org 2766 could be demonstrated in terms of rating scales or cognitive functions. PMID- 2991818 TI - beta-Endorphin cerebrospinal fluid decrease in untreated parkinsonian patients. AB - We measured CSF and plasma contents of beta-endorphin (beta-EP), beta-lipotropin (beta-LPH), and ACTH in 24 patients with Parkinson's disease; 14 had not been treated. CSF beta-EP concentrations in untreated patients were lower than in 15 controls (p less than 0.005), but values did not differ significantly in treated and untreated patients. In untreated and treated patients, ACTH and beta-LPH CSF, and beta-EP, beta-LPH, and ACTH plasma concentrations were in the same range as controls. The Parkinson's disease-related decrease of CSF beta-EP levels further supports the concept that there is a generalized brain disorder in Parkinson's disease affecting more than dopaminergic neurons in the substantia nigra. PMID- 2991819 TI - Antivirus antibodies in myasthenia gravis. AB - Serum antibodies to influenza A, measles, rubella, cytomegalovirus, varicella zoster, herpes simplex type 1, and mumps have been assayed in 104 patients with myasthenia gravis, grouped according to clinical features plus thymus pathology, and compared with matched controls. No significant differences in incidence or antibody titer were detected. In 37 patients with recent onset of symptoms, the incidence of antibody to coxsackieviruses B1-B6 was less than in controls. Juvenile-onset cases also demonstrated antibody to Epstein-Barr virus at the expected frequency. These results weaken the case for any of these common viruses, or the response to them, contributing to the pathogenesis of myasthenia gravis. PMID- 2991820 TI - Lumbosacral plexopathy in cancer patients. PMID- 2991821 TI - [Obstructive icterus caused by a biliary thrombus due to hemobilia caused by a hepatocarcinoma. Case observation]. PMID- 2991822 TI - Selective blockade of benzodiazepine receptors by Ro 15-1788 prevents foot shock induced decrease of low affinity gamma-aminobutyric acid receptors. AB - The cerebral cortex of unstressed rats has a higher density of low affinity gamma aminobutyric acid (GABA) receptors than that of stressed animals. Stress (handling or foot shock) produces a sudden decrease in the total number of low affinity GABA receptors in the cerebral cortex of unstressed rats but leaves unchanged the density of GABA receptors in the cortex of stressed animals. The in vivo administration of Ro 15-1788 (30 mg/kg per os), a specific benzodiazepine receptor antagonist, completely prevents the effect of footshock on the low affinity GABA receptors. The results suggest that (a) benzodiazepine recognition sites are involved in the action of stress on GABA receptors, and (b) stress may release an endogenous ligand for the benzodiazepine recognition site. PMID- 2991823 TI - Dopamine-sensitive adenylate cyclase in rabbit portal vein. AB - The effect of dopamine (DA) on 3',5'-cyclic adenosine monophosphate (cAMP) generation in the rabbit portal vein was investigated. Dopamine added to homogenates of rabbit portal vein increased the concentration of cAMP. DA elicited cAMP increase was exclusively inhibited by DA antagonists fluphenazine and haloperidol suggesting the existence of DA receptors (of the DA1 subtype) in the portal vein. PMID- 2991824 TI - Synaptic organization of the cerebello-thalamo-cerebral pathway in the cat. I. Projection of individual cerebellar nuclei to single pyramidal tract neurons in areas 4 and 6. AB - The neural connections of the dentate (DN) and the interpositus (IN) nuclei to the motor cortex and area 6 were investigated by recording intracellular postsynaptic potentials from fast and slow pyramidal tract neurons (PTNs) in the anesthetized cat. Localized stimulation of DN and IN produced di- or polysynaptic EPSPs in fast and slow PTNs in the "forelimb area" of the motor cortex and area 6. The effects of stimulation of the two cerebellar projections were essentially the same, although some regional difference of their relative strength was noted. In these cortical areas, the majority of fast and slow PTNs received convergent inputs from both DN and IN. By examining the interaction of DN- and IN-evoked EPSPs, spatial facilitation and occlusion at the level of the thalamus were demonstrated. Therefore, it was concluded that at least a portion of the convergence of the dentate and the interpositus inputs occurred at the level of the ventrolateral nucleus of the thalamus. PMID- 2991825 TI - Synaptic organization of the cerebello-thalamo-cerebral pathway in the cat. II. Input-output organization of single thalamocortical neurons in the ventrolateral thalamus. AB - Input-output neural organization of single thalamocortical (T-C) neurons in the ventrolateral nucleus (VL) of the thalamus was investigated using an intracellular recording technique in the anesthetized cat. Stimulation of the dentate (DN) and the interpositus (IN) nuclei produced monosynaptic unitary EPSPs of large amplitude in T-C neurons projecting to the motor cortex or area 6 over the entire mediolateral region of VL. The thalamic projections from DN and IN are very wide and there is a considerable overlap between the dentate and the interpositus projection areas in VL. And in this overlapping area, a considerable number of T-C neurons (50%) receive inputs from both DN and IN. More than 40% of T-C neurons were antidromically activated from widely separated electrodes in the motor cortex, indicating that the cortical arbolization of single T-C neurons is very wide and the number of these neurons with widely divergent projections is considerably large. PMID- 2991826 TI - Nerve growth factor enhances central synaptic function of Ia sensory neurons. AB - Daily treatment of neonatal rats with nerve growth factor (NGF) significantly enhanced monosynaptic excitatory postsynaptic potentials (EPSPs) evoked in spinal motoneurons by muscle afferent volleys. A few weeks after crush of a muscle nerve, the EPSPs elicited by afferent volleys from the muscle nerve were markedly depressed. This synaptic depression could be partly prevented by daily application of NGF. It is concluded that central synaptic function of Ia sensory neurons is responsive to exogenous NGF in postnatal animals. PMID- 2991828 TI - Altered receptors for 1,25-dihydroxycholecalciferol in vitamin D-dependent rickets. PMID- 2991827 TI - Intracellular receptors mediate the biologic action of 1,25-dihydroxyvitamin D3. PMID- 2991829 TI - Nutrition classics. Proceedings of the National Academy of Sciences of the United States of America, Volume 62, 1969. Chromosomal receptor for a vitamin D metabolite. By Mark R. Haussler and Anthony W. Norman. PMID- 2991830 TI - Acute respiratory tract infections of children in hospital: a viral and Mycoplasma pneumoniae profile. AB - Respiratory specimens and blood were collected from all infants and children admitted with acute respiratory illness to a paediatric unit in Christchurch from May to November (late autumn, winter and spring) 1983, to define the viral aetiological agents involved. A virus or Mycoplasma pneumoniae was identified in 160 (50%) of 317 children studied by the rapid indirect immunofluorescence, virus culture and/or serological techniques. Aetiological agents were detected in 71% of children with bronchiolitis, 57% with pneumonia, 53% with bronchitis, 40% with laryngotracheitis (croup), and 45% with upper respiratory tract illness. Respiratory syncytial virus was the most frequently identified virus, confirming the importance of this virus as a cause of respiratory illness requiring hospitalisation of young children in Christchurch. An epidemic due to influenza A/Dunedin/7/83 (HINI) and A/New Caledonia/4/83 (HINI) viruses occurred during the study period. PMID- 2991831 TI - Science and parascience: a modern confusion. PMID- 2991833 TI - Asymptomatic first-trimester liver cell adenoma: diagnosis by fine-needle aspiration cytology with cytochemical and ultrastructural study. AB - A large asymptomatic first-trimester liver cell adenoma was diagnosed by fine needle aspiration cytology. Therapeutic abortion was performed to reduce the rate of growth and achieve reduction in size before resection. Computed tomography scan evidence suggested that abrupt necrosis occurred after the therapeutic abortion, which might have precipitated an emergency partial hepatectomy. The tumor did not regress in size and was estrogen and progesterone receptor-assay negative. PMID- 2991832 TI - Comparison of lipid and androgen levels after conjugated estrogen or depo medroxyprogesterone acetate treatment in postmenopausal women. AB - Thirty-five women who had undergone a natural or surgical menopause were randomized to receive either 0.625 mg of conjugated estrogen to be ingested for 25 days each month or 150 mg of depo-medroxyprogesterone acetate intramuscularly every three months. Plasma lipids were determined before and after one year of therapy. Serum androgens were measured before and after six months of therapy. Conjugated estrogens and depo-medroxyprogesterone acetate had similar effects on plasma lipids. Cholesterol was decreased (P less than .02 for conjugated estrogen therapy and P less than .01 for depo-medroxyprogesterone acetate therapy), as was low-density lipoprotein cholesterol (P less than .02 for conjugated estrogen and P less than .05 for depo-medroxyprogesterone acetate). Conjugated estrogens also significantly increased high-density lipoprotein cholesterol (P less than .02). High-density lipoprotein cholesterol levels were unchanged after depo medroxyprogesterone acetate therapy. Serum androgens were generally unchanged after depo-medroxyprogesterone acetate or conjugated estrogen therapy. The data from this study suggest that long-term conjugated estrogen and depo medroxyprogesterone acetate treatment have similar effects on lipid and androgen levels except that high-density lipoprotein cholesterol was not significantly increased by depo-medroxyprogesterone acetate. PMID- 2991834 TI - Diagnosis of breast masses in pregnant and lactating women by aspiration cytology. AB - Five pregnant women with dominant breast masses underwent fine-needle aspiration cytology. Fine-needle aspiration cytology findings of the three patients with benign pregnancy adenomas and the two patients with ductal carcinoma are described. More extensive use of fine-needle aspiration cytology in the evaluation of breast masses in pregnant patients may decrease delays in diagnosis of breast cancer in pregnant women. PMID- 2991835 TI - Vaginal glomus tumor: case presentation and ultrastructural findings. AB - Vaginal glomus tumors are exceptionally rare, and have not been reported in the literature. A case of vaginal glomus tumor is described. Light and electron microscopic views are presented. The differential diagnosis and surgical approach are discussed. PMID- 2991836 TI - Malignant placental site trophoblastic tumor. AB - Placental site trophoblastic tumor is a rare variant of trophoblastic disease. The malignant form of this disease with metastasis and death is even more infrequent, as is evident from only five cases in the literature. A 30-year-old black woman who died from this disease is presented. Patients with metastases are at extremely high risk with no reported survivors and must be treated aggressively with chemotherapy and cytoreductive surgery. This type of trophoblastic tumor apparently exhibits a different biologic behavior as compared with choriocarcinoma. PMID- 2991837 TI - [Familial cumulation of the incidence of gastroenteritis caused by Rotavirus]. PMID- 2991838 TI - Reflections on the management of adenoid cystic carcinoma of the nasal cavity and paranasal sinuses. AB - Adenoid cystic carcinoma of the nasal cavity and paranasal sinuses has a unique natural history. It has a propensity for frequent local recurrence and early perineural and hematogenous spread. These factors, together with a complex anatomic situation, make adequate oncologic surgery extremely difficult. The incidence of local recurrence is lower in those patients receiving more radical primary surgery, but the choice between conservative and radical management may not alter prognosis. Indeed, it is doubtful if any patients are cured of this disease. PMID- 2991839 TI - Human papillomavirus in various lesions of the head and neck. AB - The association of human papillomavirus with benign and malignant epithelial lesions of the head and neck has been studied by a peroxidase-antiperoxidase technique having immunospecificity against genus-specific structural antigens of the papillomaviruses. More than 360 specimen blocks from 144 patients were evaluated. There was evidence of human papillomavirus antigen in three out of eight patients with childhood-onset laryngeal papillomas (37.5%) and in four out of eight patients with adult-onset papillomas (50%). A patient with an unusual flat, wartlike lesion appearing as an oral cavity leukoplakia had detectable papillomavirus antigen in it. None of the 13 cases of inverting papilloma or any of the malignant lesions studied showed evidence for the presence of papillomavirus antigen. There is currently only suggestive evidence for the oncogenic potential of human papillomavirus in the head and neck. PMID- 2991840 TI - Pleomorphic adenoma of the nasal septum. PMID- 2991841 TI - Defective regulation of immune responses in croup due to parainfluenza virus. AB - In order to determine if defects in regulation of immune responses play a role in the pathogenesis of croup, we studied 37 infants and children with either croup or upper respiratory illness alone due to parainfluenza virus (PV). PV-specific IgE responses were determined by an enzyme-linked immunosorbent assay, cell mediated immune responses to PV antigen were studied by in vitro lymphocyte transformation assays, and suppressor cell function was determined by addition of histamine to lymphocyte transformation assays. In comparison to patients with upper respiratory illness alone, patients with croup had increased production of PV-specific IgE antibody, increased lymphoproliferative responses to PV antigen, and diminished histamine-induced suppression of lymphocyte transformation responses to PV. These results suggest that a defect in suppressor function exists among croup patients. Similar defects have been demonstrated in bronchiolitis and atopic diseases, providing an immunologic link between the three illnesses. PMID- 2991842 TI - [Functional constipation]. PMID- 2991844 TI - Positive regulation of the colicin E1 gene by cyclic AMP and cyclic AMP receptor protein. AB - In previous experiments, we showed that the in vivo transcription of the colicin E1 gene was dependent on cyclic AMP in adenylate cyclase-defective mutant cells of Escherichia coli (Ebina, Y. and Nakazawa, A (1983) J. Biol. Chem. 258, 7072 7078). We now show that cyclic AMP and cyclic AMP receptor protein stimulated the in vitro transcription of the gene in the presence of spermidine. As determined in DNase I protection experiments, two binding sites for the complex of cyclic AMP and the receptor protein were identified about 60 base pairs (CRP-1) and 110 base pairs (CRP-2) upstream from the transcription initiation site of the colicin E1 gene. CRP-1 had a higher affinity for the complex than that of CRP-2. Substituting an unrelated DNA sequence for CRP-2 reduced the efficiency of in vitro stimulation of the gene by cyclic AMP and the receptor protein. These potential binding sites for the cyclic AMP-cyclic AMP receptor protein complex probably participate in the stimulation of the colicin E1 gene transcription. PMID- 2991843 TI - S1-sensitive sites in the supercoiled double-stranded form of tomato golden mosaic virus DNA component B: identification of regions of potential alternative secondary structure and regulatory function. AB - The sensitivity of the supercoiled double-stranded form of the DNA of tomato golden mosaic virus (TGMV), a geminivirus, to the single-strand specific enzyme S1 nuclease has been demonstrated. Specific S1 cleavage sites were identified in TGMV DNA component B by cloning into the single-strand bacteriophage vector M13 mp8 and sequencing of the inserted DNA. Analysis of the DNA sequence at the sites of S1 sensitivity in TGMV DNA component B revealed several possible regions of alternative secondary structure which were clustered in an intergenic region upstream of the starts of the two major open reading frames which are in opposite orientations. This region contains putative transcriptional promoter and modulatory sequences and a possible replication origin. The extreme S1 sensitivity of the supercoiled form of TGMV DNA component A precluded its cloning under the conditions employed for selective cleavage of DNA component B. PMID- 2991845 TI - Regulation of the Escherichia coli DNA topoisomerase I gene by DNA supercoiling. AB - The transcriptional control region of THE E. coli DNA topoisomerase I (topA) gene has been fused to the galactokinase (galK) gene coding region in a recombinant plasmid. In vivo synthesis of the galactokinase produced from such a plasmid has been measured and found to be reduced when mutations in the genes coding for DNA gyrase subunits are introduced into the cell or when gyrase inhibitors are present. In vitro transcription-translation of the galactokinase gene product confirms that a supercoiled DNA template is required for efficient transcription from the topA gene promoter. These results indicate that the amount of DNA topoisomerase I activity in E. coli is regulated by the extent of DNA supercoiling and can contribute to the overall modulation of DNA superhelicity and the expression of other genes. PMID- 2991846 TI - Functional analysis of the regulatory region of polyoma mutant F9-1 DNA. AB - Functional analysis of the transcriptional control region of the polyoma (Py) mutant F9-1 reveals that the mutation is located in a region of Py DNA required for at least two functions. First, an enhancer element which includes the F9-1 mutation was characterized by deletion analysis. This element, described previously as enhancer B is essential for viral early gene expression in F9 stem cells whereas enhancer A is unnecessary for transcriptional activity in embryonal carcinoma (EC) cells. Second, a CCACCC motif, present twice in the 3' part of enhancer B is also required in cis for the response to a heterologous enhancer. This suggests that a promoter element is present in this region of the polyoma genome which overlaps Py enhancer B. We also demonstrate the enhancement of the polyoma early promoter activity in F9 stem cells by MSV sequences. The significance of these observations is discussed. PMID- 2991847 TI - Alkali-labile structures linked to the 5' ends of Bacillus subtilis short DNA chains. AB - Alkali-labile portion covalently linked to the 5' ends of Bacillus subtilis short DNA chains, the putative primer RNA for discontinuous DNA synthesis, was isolated and analyzed using a temperature sensitive DNA polymerase I mutant, which accumulates nascent DNA fragments at a restrictive temperature. A novel oligonucleotide structure as well as mono- to triribonucleotide stretches were isolated at the 5' end of the short DNA chains. A structure for the novel oligonucleotide is proposed to be p5' X3' pp5' rN, where X represents unidentified nucleoside with a peculiar property. Possible metabolic relationship between these molecules and primer RNA has been discussed. PMID- 2991848 TI - Nucleotide sequence of transforming human c-sis cDNA clones with homology to platelet-derived growth factor. AB - Three c-sis cDNA clones were obtained from polyadenylated RNA of a human T-cell lymphotropic virus (HTLV) type I transformed cell line. Two clones, designated pSM-1 and pSM-2, have cDNA inserts of 2498 and 2509 base pairs (bp), respectively, excluding the sizes of the guanylate tails, and the polyadenylate tracts. These clones are shorter than the estimated size of the c-sis mRNA of 4200 bp. Both of these clones can transform NIH 3T3 cells. The third clone, designated pSM-3 has a cDNA insert of 1421 bp and lacks transforming activity. The sequence of clone pSM-1 reveals a single long open reading frame (nucleotides 118-840) encoding chain A of platelet-derived growth factor, and two segments with homology to v-sis (nucleotides 182-871 and 1021-1325). Sequence homology is noted in the 3' untranslated region to the corresponding regions of the beta 1 interferon (IFN), human and murine beta-nerve growth factor (NGF), human interleukin 2 (IL2) genes, and tubulin pseudogenes. However, no typical AATAAA polyadenylation signal is present. An alternating (dCdA)n X (dGdT)n sequence is present in the 3' flanking cellular sequences similar to those in the corresponding position of the human proenkephalin gene, in the first intron of the gamma-IFN gene, and the second intron of the beta-NGF gene. PMID- 2991850 TI - Ordered processing of Escherichia coli 23S rRNA in vitro. AB - In an RNase III-deficient strain of E. coli 23S pre-rRNA accumulates unprocessed in 50S ribosomes and in polysomes. These ribosomes provide a substrate for the analysis of rRNA maturation in vitro. S1 nuclease protection analysis of the products obtained in in vitro processing reactions demonstrates that 23S rRNA processing is ordered. The double stranded stem of 23S rRNA is cleaved by RNase III in vitro to two intermediate RNAs at the 5' end and one at the 3' end. Mature termini are then produced by other enzyme(s) in a soluble protein fraction from wild-type cells. The nature of the reaction at the 5' end is not clear, but the reaction at the 3' end is exonucleolytic, producing three heterogeneous mature termini. The two reactions are coordinated; 3' end maturation progresses concurrently with cleavages at the 5' end. Two results suggest a possible link between final maturation and translation: in vitro, mature termini are formed efficiently in the presence of additives required for protein synthesis; and all the processing intermediates detected from in vitro reactions are also found in polysomes from wild-type cells. PMID- 2991849 TI - Yeast contains two functional genes coding for ribosomal protein S10. AB - The DNA sequence of the second copy of the gene coding for yeast ribosomal protein S10 was determined and compared with the sequence of the first gene-copy. In addition, the sites at which the transcription of these genes start and terminate are identified. The amino acid coding regions of the two gene copies are virtually identical. The leader and in particular the trailer sequences, however, are significantly different, while the intervening sequences have hardly any homology. Taking advantage of the sequence differences we could establish that both genes are expressed in the vegetatively growing yeast cell; the respective transcripts, however, differ in their relative amounts. PMID- 2991851 TI - Transposition of a long member of the L1 major interspersed DNA family into the mouse beta globin gene locus. AB - A long member of the highly repeated long interspersed DNA family L1Md (for L1 in Mus domesticus) has integrated by transposition into a target site which lies between the two adult beta globin genes of mouse. DNA hybridization and nucleotide sequence analysis show that this target site, which is part of the single copy DNA flanking the globin genes, is interrupted by the L1 element in one chromosome but is uninterrupted in both allelic and ancestral chromosomes. Other large DNA rearrangements of the region between the two adult beta globin genes are also associated with these allelic chromosomes, and include insertions or deletions of both single copy DNA and simple and complex repetitive DNA. This has caused extensive reorganization of this intergenic region. However, the distance between the two genes flanking this region remains conserved, suggesting that the spacing of the globin genes may be subject to conservative selection. PMID- 2991852 TI - Chromosomal rearrangements associated with LINE elements in the mouse genome. AB - Two segments of DNA that have apparently inserted in the interval between the two adult beta-globin genes in BALB/c (Hbbd haplotype) but not in C57B1/10 (Hbbs haplotype) mouse strains have been described (1). These putative insertions, each about 1000 bp in length, mapped near a repetitive element. To determine the precise position of these alleged insertions, their target sites, and the nature of their boundaries, we cloned and sequenced the appropriate regions of both chromosomes. One of the two segments is not an insertion but rather a region between two independently integrated L1 repetitive elements (LINEs) (2), one in Hbbd and the other in the Hbbs chromosome. The other segment is an insertion of 940 bp which is located within the L1 element in the Hbbd chromosome. This insert is unusual in that it exists in only one copy in the BALB/c genome. PMID- 2991853 TI - Analysis of the regulatory region of the ssb gene of Escherichia coli. AB - The regulation of the ssb gene of E. coli has been studied. We reported earlier that the SOS box of the neighbouring uvrA gene also controls the transcription of the ssb gene. Detailed analysis of the upstream region of ssb by S1 mapping reveals the existence of three in vivo functional promoters of which the most upstream one (PI) is inducible by DNA damage. Measurement of galactokinase synthesis using galK fusion plasmids indicates that the uninduced level of transcription from the PI promoter is low. Ssb multicopy plasmids lacking the PI promoter still complement the UV sensitivity of an Ssb mutant. The role of the three promoters in the regulation of the level of Ssb protein in the cell, is discussed. PMID- 2991854 TI - Inhibition of the herpes simplex virus thymidine kinase gene transfection in Ltk- cells by potential Z-DNA forming polymers. AB - It has been demonstrated that certain alternating purine and pyrimidine sequences may assume a left-handed Z-DNA conformation. In order to evaluate the possibility that Z-DNA is involved in the modulation of gene expression, we examined the ability of various synthetic DNA polymers to affect the transfection of herpes simplex virus thymidine kinase (HSVtk) gene in Ltk- cells using the DNA-calcium phosphate cotransfection technique. We found that potential Z-DNA forming polymers such as, poly(dG-m5dC) X poly(dG-m5dC) and poly(dG-dC) X poly(dG-dC), cotransfected with the tk gene decreased the level of Tk+ transformed colonies. In contrast, cotransfection of the tk gene with polymers which do not assume Z conformation such as, poly(dG) X poly(dC) or poly(dA-dT) X poly(dA-dT) showed no effect on the number of colonies formed. About 50% inhibition of the Tk+ colony formation was obtained by 0.4 micrograms of poly(dG-m5dC) X poly(dG-m5dC), or by 2 micrograms of poly(dG-dC) X poly(dG-dC). DNA uptake into Ltk- cells was not significantly affected by any of these polymers. Approximately 20-42 base pairs (bp) long alternating dG-dC sequence linked at either the 5'-end or 3'-end of tk gene were cloned into plasmids. These recombinant plasmids, however, showed no remarkable effect upon the transfection of Ltk- cells. The DNAs of Tk+ colonies obtained by transfecting these recombinant plasmids were digested with BssH II and analyzed by Southern blotting. We demonstrated that the dG-dC sequences proximal to the tk gene were integrated into cellular DNA. All the presented results indicate that only larger polymers with the potential to assume a Z-DNA conformation may affect tk gene transfection either by inhibiting transcription or more probably by affecting the stable integration of the tk gene into the host chromosome. PMID- 2991857 TI - Adenovirus sequences required for replication in vivo. AB - We have studied the in vivo replication properties of plasmids carrying deletion mutations within cloned adenovirus terminal sequences. Deletion mapping located the adenovirus DNA replication origin entirely within the first 67 bp of the adenovirus inverted terminal repeat. This region could be further subdivided into two functional domains: a minimal replication origin and an adjacent auxillary region which boosted the efficiency of replication by more than 100-fold. The minimal origin occupies the first 18 to 21 bp and includes sequences conserved between all adenovirus serotypes. The adjacent auxillary region extends past nucleotide 36 but not past nucleotide 67 and contains the binding site for nuclear factor I. PMID- 2991855 TI - An unusually long poly(purine)-poly(pyrimidine) sequence is located upstream from the human thyroglobulin gene. AB - A region of human genomic DNA encompassing the 5' end of the thyroglobulin gene has been sequenced and the position of the transcriptional start site has been determined. The 5' non-translated portion of the mRNA displays a quasi palindromic sequence which could allow this region to adopt a hairpin structure. The first exon of the gene encodes a 19 amino-acids signal peptide and the 3 first amino acids of the mature protein. Apart from the canonical TATA-Box and from a CAAT-Box homology, the promoter region contains a 209 bp-long poly(purine) poly (pyrimidine) sequence located between positions-512 and -304 relative to the transcription start. When contained in a supercoiled plasmid, this sequence exhibits sensitivity to S1 nuclease at two distinct positions. A precise mapping of the borders of the sensitive regions was achieved by extending primers from both ends of the sequence after digestion by the enzyme. The resulting data can be explained by a model involving the formation of a triple helix structure. PMID- 2991856 TI - Are there insertions in the ribosomal DNA of vertebrates? AB - We have analysed the Eco RI restriction pattern of rDNA of the newt Triturus vulgaris and of some other amphibian species by Southern blotting and hybridization with nick-translated Xenopus rDNA prepared from the recombinant plasmids pXlr11 and pXlr12 (21). After hybridization with r11, the 28S coding fragments become visible in two bands, a prominent one of 5.3 kb and a weak band of 5.9 kb representing about 8% of the 28S genes. The evidence obtained so far by additional digestions with Bam HI and Bgl II indicates that in this species and in Triturus helveticus the coding regions of the 5.9 kb fragments are interrupted by an insertion 0.6 kb in length located in a 1.6 kb Bgl II fragment at the 3' end of the Eco RI fragment, which we believe to be the first described in a vertebrate. PMID- 2991858 TI - Trans acting regulation of beta globin gene expression in erythroleukemia (K562) cells. AB - K562 cells are induced by hemin to produce gamma and epsilon globin but not beta globin, although the beta globin gene is intact, and when isolated is expressed in a transient expression assay (1, 2). We have previously shown that an epsilon globin gene transferred into K562 cells is expressed and inducible (3). In this paper, we report the stable transfer of a sickle or betaS globin gene into K562 cells. Thirty-six different transformed lines were tested; 24 of 36 lines contained an intact betaS globin gene. However, using S1 nuclease, Dot blot, and Northern blotting analyses, none of these lines showed beta globin mRNA expression. These results indicate that trans acting factors are responsible for the lack of expression of the beta globin gene in K562 cells. PMID- 2991859 TI - The structure and evolution of a 461 amino acid human protein C precursor and its messenger RNA, based upon the DNA sequence of cloned human liver cDNAs. AB - Human liver cDNA coding for protein C has been synthesized, cloned and sequenced. The abundance of protein C message is approximately 0.02% of total mRNA. Three overlapping clones contain 1,798 nucleotides of contiguous sequence, which approximates the size of the protein's mRNA, based upon Northern hybridization. The cDNA sequence consists of 73 5'-noncoding bases, coding sequence for a 461 amino acid nascent polypeptide precursor, a TAA termination codon, 296 3' noncoding bases, and a 38 base polyadenylation segment. The nascent protein consists of a 33 amino acid "signal", a 9 amino acid propeptide, a 155 amino acid "light" chain, a Lys-Arg connecting dipeptide, and a 262 amino acid "heavy" chain. Human protein C and Factor IX and X precursors possess about one third identical amino acids (59% in the gamma-carboxyglutamate domain), including two forty-six amino acid segments homologous to epidermal growth factor. Human protein C also has similar homology with prothrombin in the "leader", gamma carboxyglutamate and serine protease domains, but lacks the two "kringle" domains found in prothrombin. PMID- 2991860 TI - Human N-ras: cDNA cloning and gene structure. AB - The structure and organisation of the human N-ras gene has been determined by analysing cDNA clones derived from the two main mRNA transcripts. One clone in particular is 4.1 Kb long and originates from the larger (4.3 Kb) message. Sequence analysis of this clone has revealed that the N-ras gene consists of seven exons. A second clone deriving from the smaller (2 Kb) message shows that the difference between the two transcripts is a simple extension through the termination site of the 2 Kb transcript. Using S1 analysis, two transcriptional starts have been mapped, 10 bp apart. There is no obvious TATA box in the expected promoter region of the gene, though there are 4 GGGCGG sequences surrounding the start sites. The 5' untranslated sequence contains 2 ATGs upstream of the initiation codon. PMID- 2991862 TI - Adenovirus E3-early promoter: sequences required for activation by E1A. AB - To identify sequences within the adenovirus-5 E3 promoter necessary for E1A trans activation, a series of promoter deletion mutants were constructed and analysed. A region between positions -82 and -105 was shown to be critical both for E1A induced expression as well as uninduced expression. The importance of this region was confirmed by constructing hybrid promoters consisting of E3 and Herpes simplex virus thymidine kinase sequences. The E1A insensitive tk promoter could be converted to an E1A sensitive promoter by replacing sequences upstream of position -79 with the corresponding region of the E3 promoter. This critical region of the E3 promoter contains a sequence 5' AGATGACTA3' which is also present in important upstream regions of the E2A and E4 promoters. PMID- 2991861 TI - Structural features of the hisT operon of Escherichia coli K-12. AB - The DNA sequence of a 2,3-kilobase segment of the E. coli hisT operon was determined. Analysis of the sequence indicated that the upstream gene in the operon encodes a 36,364-dalton polypeptide, which runs aberrantly on SDS polyacrylamide gels. The distal hisT gene encodes the tRNA modification enzyme, pseudouridine synthase I, which was shown to have a polypeptide molecular mass of 30,399 daltons. The DNA sequence was consistent with the phenotypes and hisT expression of mutant operons. Analysis of the sequence and genetic complementation experiments demonstrated that the upstream and hisT genes are evolutionarily, structurally, and functionally unrelated; however, translation signals for the two genes overlap, which is consistent with genetic evidence suggesting translational coupling. Codon usage in the upstream gene is radically different from the hisT gene and may underlie the differential expression observed from the operon. Gene-inactivation experiments and S1-mapping of in vivo transcripts indicated that the operon contains an additional upstream gene. S1 mapping experiments also confirmed the presence of an internal promoter, which might be stringently controlled. Taken together, these results show that the structure of the hisT operon is complex and suggest that the operon might be regulated at several levels. PMID- 2991863 TI - BamHI RFLP linked to the human urokinase gene. PMID- 2991864 TI - [Coexistence of lung cancer with other malignant neoplasms]. PMID- 2991865 TI - Fenoterol: a beta2-adrenergic agonist for use in asthma. Pharmacology, pharmacokinetics, clinical efficacy and adverse effects. AB - Fenoterol (hydroxyphenylorciprenaline) is chemically closely related to metaproterenol (orciprenaline). It has a higher bronchodilating potency than metaproterenol, albuterol (salbutamol in Europe) or terbutaline. The beta 2 selectivity of fenoterol at normal oral and inhaled doses is the same as for albuterol and terbutaline. Its pharmacodynamic effects are similar to those of other selective beta 2-adrenoceptor agonists. It has a high first-pass metabolism. The long half-life previously reported in the literature (7 hours) is mainly the half-life of inactive fenoterol metabolites. The duration of action at equipotent bronchodilating doses seems to be the same as for albuterol and terbutaline, and not longer, as previously reported. Inhalation of beta adrenoceptor agonists is the superior route of administration. Side effects do not usually occur at normal therapeutic doses. One puff of fenoterol (200 micrograms) is about equipotent to 2 puffs of albuterol (2 X 100 micrograms) or 2 puffs of terbutaline (2 X 250 micrograms) with the same duration of effect. In patients who overdose with the metered-dose inhaler (MDI), side effects occur at half the number of puffs with fenoterol. Dosage for an acute attack in children is 1 puff (200 micrograms), repeated within 5 minutes if necessary; in adults 1-3 puffs can be given. For maintenance therapy, the dose in adults is 1-2 puffs 2-4 times daily, while in children 1 puff at night and 1 in the morning may be sufficient. The usual oral dosage has been 5-10 mg 3 times daily. PMID- 2991866 TI - [Function and ultrastructure of peripheral blood neutrophils in a patient with granulocytic myeloperoxidase deficiency]. PMID- 2991867 TI - Increased susceptibility to infection in hypothermic children: possible role of acquired neutrophil dysfunction. AB - The addition of hypothermia to regimens to control cerebral edema in children at our institution has been associated with a substantial incidence of infectious complications. Of the 13 children maintained at 30 degrees C to prevent cerebral edema, 3 developed Haemophilus influenzae pneumonia and 2 developed Streptococcus pneumoniae sepsis (one with pneumonia). The importance of neutrophil (PMN) function for elimination of bacterial pathogens prompted in vitro studies of PMN function at clinically attainable hypothermic temperatures. Neutrophils at 30 degrees C had significantly less ability to migrate towards a chemotactic stimulus (45 +/- 10% inhibition; P less than 0.02), to ingest staphylococci (22 +/- 5% inhibition; P less than 0.01) and to be metabolically activated as measured by superoxide production (35 +/- 10% inhibition; P less than 0.01) or by chemiluminescence (18 +/- 8% inhibition; P less than 0.05). These in vitro findings support the clinical observation that persons with decreased body temperature may be at an increased risk for bacterial infections secondary to PMN dysfunction. PMID- 2991868 TI - Prevention of pertussis, Haemophilus and varicella infections. PMID- 2991869 TI - Recurrence of synovial sarcoma of the abdomen in a 10-year old child. AB - The case of a 10 year old child with a synovial sarcoma of the left iliac fossa, which recurred 12 months after simple large surgical excision, is reported here. No related lesions and, in particular, no lesion of the articular capsule of the left hip could be found, either on X-ray or at the time of the two excisions. Microscopically, the tumor showed, in all the areas examined, the existence of two characteristic morphological aspects, associating epithelial like type cells and elongated fusiform cells. These features correspond exactly to the classic "biphasic" type of synovial sarcoma. This location of a synovial sarcoma is an uncommon one, both in children and in adults. PMID- 2991870 TI - Primary hepatocellular carcinoma associated with Wilson's disease in a young woman. AB - A 27 year old woman with hepato-lenticular degeneration (Wilson's disease) was found to have primary hepatocellular carcinoma (PHC) three and a half years after she was started on treatment with D-penicillamine. The tumour was resected since when she has remained well. Her liver function tests were normal throughout the course of her disease. The available literature is reviewed and possible mechanisms for this association proposed. PMID- 2991871 TI - Isolated adrenocorticotrophic hormone (ACTH) deficiency associated with acute adrenal crisis. AB - A 37 year old black female presented with congestive cardiac failure, 2 months postpartum. She developed spontaneous hypoglycaemia and symptoms of acute adrenal crisis (hypotension, nausea, abdominal pain and tachycardia with small thready pulse), which responded to i.v. dextrose, sodium chloride and hydrocortisone. Biochemical investigations revealed low serum cortisol and plasma adrenocorticotrophic hormone (ACTH) levels. The patient initially showed an impaired cortisol response to intramuscular aqueous tetracosactrin, but an exuberant response after priming with intramuscular tetracosactrin depot. These findings, together with the normal remaining pituitary function, led us to conclude that this patient had isolated ACTH deficiency associated with congestive cardiac failure and acute adrenal crisis. PMID- 2991873 TI - Acromegaly presenting as subarachnoid haemorrhage in a 76 year old woman. AB - Acromegaly usually presents in middle age with the gradual onset of classical signs and symptoms. We report the case of an elderly woman who presented with subarachnoid haemorrhage due to infarction of a pituitary adenoma and rupture into the subarachnoid space. PMID- 2991872 TI - Hypereosinophilic syndrome presenting as peripheral neuropathy. AB - An unusual case of hypereosinophilic syndrome is described which presented with peripheral neuropathy with no evidence of cardiac involvement. The response to steroid therapy is documented and the literature on peripheral neuropathy in hypereosinophilic syndrome is reviewed. PMID- 2991874 TI - Malignant inflammatory fibrous histiocytoma of bone and acute lymphoblastic leukaemia. AB - A patient is described who developed a malignant inflammatory fibrous histiocytoma of vertebral bone (MFH) 4 months after the onset of acute lymphoblastic leukaemia. This is the first report of the simultaneous occurrence of these two conditions. Similarities to the more frequent occurrence of another histiocytic malignancy, histiocytic medullary reticulosis, following acute lymphoblastic leukaemia, are noted. The usefulness of gallium scanning to monitor the response of this tumour to treatment is demonstrated. PMID- 2991875 TI - Acute pancreatitis in association with small cell lung carcinoma: potential pitfall in diagnosis and management. AB - Tumour metastases to the pancreas are a rare but recognized cause of acute pancreatitis, there is a 24-40% incidence of pancreatic involvement from small cell lung cancer in autopsy series but only a very few cases of tumour-induced acute pancreatitis have been described. Chemotherapy has been advocated as the primary therapy in patients with known oat cell carcinoma who develop acute pancreatitis. We describe 2 patients with acute haemorrhagic pancreatitis in association with disseminated small cell carcinoma but without evidence of tumour invasion in the gland and with gall stones present in the gall bladder. Chemotherapy would have been inappropriate therapy for these patients. PMID- 2991876 TI - Coexistent ulcerative colitis and Crohn's disease. AB - This case report describes a patient with chronic ulcerative colitis and epithelial dysplasia of 17 y duration ultimately complicated by colonic carcinoma. At laparotomy, clinically unsuspected but typical Crohn's disease of the terminal ileum was also found. The macroscopic and histological features of the resected terminal ileum were characteristic of Crohn's disease. PMID- 2991877 TI - Effect of carbonated water on growth performance of cockerels subjected to constant and cyclic heat stress temperatures. AB - Three growth trials were performed to determine the effect of carbonated water on growth performance of cockerels subjected to heat stress temperatures. In Trial 1, a 2 X 2 X 2 factorial design was used to test growth performance of Columbian crossbred cockerels between 8 and 11 weeks of age. The birds were subjected to either cyclic (day-night) heat stress (H) temperature (29 to 34 C) or cyclic (day night) moderate (M) temperature (25 to 29 C), fed either a corn soy grower (G) diet or a 20% alfalfa diet (A), and provided with tap (TW) or carbonated (CW) drinking water. In Trial 2, a 2 X 4 X 2 factorial was used to access the effect of CW on growth performance of Hubbard cockerels between 4 and 7 weeks of age. Birds were grown in the M or H cyclic thermal environments with dietary treatments consisting of A, G, A plus 1% sodium bicarbonate (AB) and G plus 1% sodium bicarbonate (GB). In Trial 3, a 2 X 2 factorial was used to test the effect of CW on growth performance of Hubbard cockerels fed the G diet and subjected to either constant heat stress (33 C) or thermoneutral (25 C) temperatures. A 24-hr photoperiod was used in each experiment. In all three trials, heat stress reduced (P less than .05) average daily gain (ADG), feed intake, and feed efficiency (G/F).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991879 TI - [Is the metyrapone test no longer useful in the evaluation of anterior pituitary corticotropin deficiency?]. PMID- 2991878 TI - Linkage of human chorionic gonadotrophin and placental lactogen biosynthesis to trophoblast differentiation and tumorigenesis. AB - Normal trophoblast of the human placenta elaborates at least two major protein hormones, chorionic gonadotrophin (hCG) and placental lactogen (hPL). Molar and choriocarcinoma tissues characteristically synthesize large amounts of hCG and hPL. To examine the role of trophoblast differentiation in the expression of the hCG and hPL genes, we studied the cytological distribution of their mRNAs in tissue sections of human hydatidiform mole and choriocarcinoma by in situ hybridization. Histologically, these tissues are in different stages of cellular differentiation. In normal placenta, hCG alpha/beta mRNA can be localized to some cytotrophoblasts and primarily to the syncytium, whereas hPL mRNA appears only in the syncytial layer. In hydatidiform mole, which still retains placental villous morphology, the hPL gene and hCG alpha and beta genes are expressed but are poorly localized because of the admixture of cyto- and syncytiotrophoblasts. By contrast, choriocarcinoma, which is devoid of placental villous pattern but in which the cyto- and syncytiotrophoblast-like components are distinguishable, expresses hCG alpha and beta in the syncytial-like areas but little, if any, hPL. These results suggest that a certain level of trophoblast differentiation, such as villous formation, is associated with hPL expression, while the hCG alpha gene and the hCG beta gene can be expressed in more disorganized tissues which contain cytotrophoblastic elements. PMID- 2991880 TI - [Infection by the LAV virus and the acquired immunodeficiency syndrome]. PMID- 2991881 TI - Inactivation of S-adenosyl-L-homocysteine hydrolase by cAMP results from dissociation of enzyme-bound NAD+. AB - S-Adenosyl-L-homocysteine hydrolase (EC 3.3.1.1) is inactivated by cAMP and also by 2'-deoxyadenosine, and in both cases, activity is restored by incubating the inactivated enzyme with NAD+. We have previously presented evidence that, despite these similarities, inactivation by these two ligands proceeds by different mechanisms. We have now used a fluorescence technique to quantitate enzyme-bound NAD+ and NADH on S-adenosyl-L-homocysteine hydrolase from Dictyostelium discoideum, and we have confirmed that cAMP and 2'-deoxyadenosine inactivate by different mechanisms. Whereas inactivation by 2'-deoxyadenosine is due to reduction of the enzyme-bound NAD+ to NADH, incubation of S-adenosyl-L homocysteine hydrolase with cAMP results in dissociation of the enzyme-bound NAD+. The dissociation is reversible, and reactivation likely occurs by restoration of the initial NAD+ content. This reversible inactivation by cAMP may be a mechanism of controlling biological methylation reactions by adjusting intracellular concentrations of S-adenosyl-L-homocysteine through action of S adenosyl-L-homocysteine hydrolase. PMID- 2991882 TI - Gene expression and cAMP. AB - By comparing the 5'-flanking region of the porcine gene for the urokinase form of plasminogen activator with those of other cAMP-regulated genes, we identify a 29 nucleotide sequence that is tentatively proposed as the cAMP-regulatory unit. Homologous sequences are present (i) in the cAMP-regulated rat tyrosine aminotransferase, prolactin, and phosphoenolpyruvate carboxykinase genes and (ii) 5' to the transcription initiation sites of cAMP-regulated Escherichia coli genes. From this we conclude that the expression of cAMP-responsive genes in higher eukaryotes may be controlled, as in E. coli, by proteins that form complexes with cAMP and then show sequence-specific DNA-binding properties. The complex formed by cAMP and the regulatory subunit of the type II mammalian protein kinase might be one candidate for this function. Based on several homologies we suggest that this subunit may have retained both the DNA-binding specificity and transcription-regulating properties in addition to the nucleotide binding domains of the bacterial cAMP-binding protein. If this were so, dissociation of protein kinase by cAMP would activate two processes: (i) protein phosphorylation by the catalytic subunit and (ii) transcription regulation by the regulatory subunit. PMID- 2991883 TI - Regulatory components in Citrobacter freundii ampC beta-lactamase induction. AB - Citrobacter freundii encodes an inducible chromosomal beta-lactamase similar to the constitutively expressed ampC beta-lactamase of Escherichia coli. In the latter species the ampC gene is located next to the fumarate reductase (frd) operon, whereas in C. freundii the ampC gene is known to be separated from frd by 1100 base pairs. This intervening DNA segment carries a gene, ampR, coding for a 31-kilodalton polypeptide. The cloned C. freundii OS60 ampC gene is inducible by beta-lactam antibiotics in E. coli, but only in the presence of an intact ampR gene. In the absence of inducer the AmpR protein represses C. freundii ampC synthesis 2.5-fold. Addition of beta-lactams induced expression from the cloned ampC beta-lactamase gene 11-fold. Thus, the AmpR protein has a positive effect on ampC expression in the presence of inducing beta-lactams. Two spontaneous mutants of C. freundii were isolated that constitutively overproduce the ampC beta lactamase. The mutations in both these strains occurred outside the frd-amp region, suggesting that there is at least one additional component in the regulatory system. With the cloned C. freundii ampC gene in E. coli, mutants with the same phenotype could be obtained. These mutations were located on the E. coli chromosome. The constitutive beta-lactamase overproduction in these mutants requires the presence of an intact ampR gene. PMID- 2991884 TI - Mutation of NH2-terminal glycine of p60src prevents both myristoylation and morphological transformation. AB - p60src, the transforming protein kinase of Rous sarcoma virus, contains the 14 carbon saturated fatty acid, myristic acid, linked through an amide bond to the alpha-amino group of its NH2-terminal glycine residue. Myristic acid is known to be attached to four other eukaryotic proteins. In each case the fatty acid is also linked through an amide bond to an NH2-terminal glycine. We have used oligonucleotide-directed mutagenesis to examine the amino acid specificity of the enzyme that myristoylates the NH2 terminus of these proteins. Replacement of the NH2-terminal glycine in p60src with either alanine or glutamic acid prevented myristoylation completely. This indicates that the myristoylating enzyme may have an absolute specificity for glycine. Strikingly, neither nonmyristoylated mutant src protein induced morphological transformation of infected cells, even though wild-type levels of phosphorylation of cellular proteins on tyrosine were observed in these cells. Since conversion of the NH2-terminal residue from glycine to alanine should have little effect on the conformation of p60src, the inability of this mutant p60src protein to induce morphological transformation suggests that the myristoyl moiety is essential for the transforming activity of the protein. PMID- 2991885 TI - Regulation of aspartate transcarbamoylase synthesis in Escherichia coli: analysis of deletion mutations in the promoter region of the pyrBI operon. AB - The catalytic and regulatory polypeptide chains of Escherichia coli aspartate transcarbamoylase are encoded by the pyrB and pyrI genes, respectively, which constitute a single transcriptional unit in the pyrBI operon. The DNA sequence immediately preceding the first structural gene, pyrB, contains a short open reading frame that could encode a 44-amino acid leader peptide and a (G+C)-rich region of dyad symmetry followed by eight thymidine residues. Synthesis of the enzyme is negatively controlled at the level of transcription depending on the cellular level of UTP, and an attenuation mechanism has been proposed to account for the 70-fold increase in pyrBI expression on pyrimidine starvation. The potential role of the dyad and eight thymidines as an attenuator was tested with a plasmid containing the promoter region of the pyrBI operon upstream of the galK coding sequence. When cells containing this plasmid, pPYRB10, were grown in a medium low in uracil, there was an 83-fold increase in galactokinase activity compared with the same cells grown at high uracil levels. This regulation is similar to that for aspartate transcarbamoylase synthesis in cells depleted of pyrimidines. Deletions constructed in the promoter region of pPYRB10 from the 3' side produced one plasmid that retained normal control of galK expression and five that exhibited greatly reduced regulation. Nucleotide sequence determination showed that the one deletion mutation that was functionally similar to the wild type plasmid contained the entire region of dyad symmetry, including the eight thymidines. The plasmids with more extensive deletions lacked the region with dyad symmetry and the eight thymidines. One of the deletion mutants that exhibited very low levels of regulation lacks the entire sequence coding for the putative leader peptide up to the major promoter. The results demonstrating the crucial role of a 19-nucleotide sequence (from -33 to -15) support an attenuation model but indicate that other mechanisms also contribute to the regulation of the pyrBI operon. PMID- 2991886 TI - Translation activates the paused transcription complex and restores transcription of the trp operon leader region. AB - It has been proposed that RNA polymerase pausing in the leader region of the tryptophan (trp) operon of Escherichia coli is responsible for the synchronization of transcription and translation essential to attenuation control. In this report we use an in vitro coupled transcription/translation system to study the effect of trp leader peptide synthesis on RNA polymerase pausing in the trp leader region. Wild-type and translation-defective trp leader templates of E. coli and Serratia marcescens were employed, and pause RNA synthesis and paused complex release (activation) were quantified relative to synthesis of the terminated leader transcript. It was observed that pausing in the trp leader region was prolonged when translation of the leader transcript was reduced by mutations in the leader region or by addition of the translation inhibitor kasugamycin or chloramphenicol. Experiments with S-30 extracts from a mutant strain that is inefficient in translating the tryptophan codons in the leader transcript indicated that ribosome movement to these codons also releases the paused transcription complex. These findings indicate that the paused trp leader transcription complex resumes transcription when released by ribosome movement over the leader peptide coding region. This release would facilitate the coupling of transcription and translation essential to attenuation control. PMID- 2991887 TI - The nucleotide sequence of the gene for human protein C. AB - A human genomic DNA library was screened for the gene for protein C by using a cDNA probe coding for the human protein. Three different overlapping lambda Charon 4A phage were isolated that contain inserts for the gene for protein C. The complete sequence of the gene was determined by the dideoxy method and shown to span about 11 kilobases of DNA. The coding and 3' noncoding portion of the gene consists of eight exons and seven introns. The eight exons code for a preproleader sequence of 42 amino acids, a light chain of 155 amino acids, a connecting dipeptide of Lys-Arg, and a heavy chain of 262 amino acids. The preproleader sequence and the connecting dipeptide are removed during processing, resulting in the mature protein composed of a heavy and a light chain held together by a disulfide bond. The heavy chain also contains the catalytic region for the serine protease. Two Alu sequences and two homologous repeats of about 160 nucleotides were found in intron E. The seven introns in the gene for protein C are located in essentially the same positions in the amino acid sequence as the seven introns in the gene for human factor IX, while the first three introns in protein C are located in the same positions as the first three in the gene for human prothrombin. PMID- 2991888 TI - Specialized nucleoprotein structures at the origin of replication of bacteriophage lambda: complexes with lambda O protein and with lambda O, lambda P, and Escherichia coli DnaB proteins. AB - The O protein of bacteriophage lambda is required for initiation of DNA replication at the lambda replicative origin designated ori lambda. The binding sites for O protein are four direct repeats, each of which is an inverted repeat. By means of electron microscopy, we have found that phage lambda O protein utilizes these multiple binding sites to form a specific nucleoprotein structure in which the origin DNA is inferred to be folded or wound. The phage lambda O and P proteins and host DnaB protein interact at ori lambda to generate a larger structure than that formed by O protein alone; P and DnaB proteins fail to form any observable complex when O protein is excluded from the reaction mixture. We conclude that the specialized nucleoprotein structure formed by phage lambda O protein and ori lambda provides for localized initiation of DNA replication by serving as the foundation for the assembly of the initial priming structure. Specialized nucleoprotein structures may be a general means to confer exceptional accuracy on DNA transactions requiring extraordinary precision. PMID- 2991889 TI - cAMP-dependent phosphorylation of bovine lens alpha-crystallin. AB - This communication reports that the A1 and B1 chains of bovine lens alpha crystallin are phosphorylated. The conclusion is based on the following evidence: (i) When soluble preparations from lens cortex are incubated with [gamma-32P]ATP, a cAMP-dependent labeling of a high molecular weight protein is obtained. (ii) After NaDodSO4/PAGE, the label is found in two bands with Mr 22,000 and 20,000, corresponding to the B and A chains of alpha-crystallin, respectively. (iii) Isoelectric focusing indicates that the radioactivity is almost exclusively in bands with pI values of 5.58 and 6.70, corresponding to the A1 and B1 chains, respectively. (iv) Similar results are obtained in experiments of [32P]orthophosphate incorporation in lens organ culture. (v) Analyses of the digested protein indicate the label is exclusively in phosphoserine. (vi) 31P NMR analyses of native, proteolytically digested, and urea-treated alpha-crystallin gives a chemical shift of 4.6 ppm relative to 85% H3PO4 at pH 7.4, suggesting that the phosphate is covalently bound to a serine in the protein. An abundance of approximately one phosphate per four or five monomer units was found. (vii) Similar results were obtained by chemical analyses of independently prepared alpha-crystallin samples. The results are consistent with the view that the A1 and B1 chains arise as result of the phosphorylation of directly synthesized A2 and B2 polypeptides. It is suggested that this metabolically controlled phosphorylation may be associated with the terminal differentiation of the lens epithelial cell and the intracellular organization of the lens fiber cell. PMID- 2991890 TI - Tetrahymena thermophila glutamine tRNA and its gene that corresponds to UAA termination codon. AB - Nucleotide sequence analysis of one of several tRNA genes cloned from Tetrahymena thermophila macronuclear DNA indicated that it corresponds to a tRNA species having TTA as anticodon. Subsequently, the tRNA species corresponding to that gene was isolated and its nucleotide sequence was determined by post-labeling techniques. The nucleotide sequence was found to be pG-G-U-U-C-C-A-U-A-m2G-U-A psi-A-G-D-G-G-D- D-A-G-U-A-C-U-G-G-G-G-A-Cm-U-Um-U-A-i6A-A-psi-C-C-C-U-U-G-A-C- m5C-U-G-G-G-U-psi-C-G-m1A-A-U-C-C-C-A-G-U-G-G-G-A-C-C-U-C-C-AOH. This tRNA sequence exactly matched the DNA sequence of the corresponding tRNA gene. The first position of anticodon is 2'-O-methyluridine (Um), forming UmUA as the anticodon, which presumably recognizes the ochre termination codon UAA. This tRNA species is aminoacylated with glutamine by a Tetrahymena crude aminoacyl tRNA synthetase fraction, suggesting that ochre termination codon is used as a glutamine codon during cytoplasmic protein synthesis in Tetrahymena. PMID- 2991891 TI - Expression of a cloned denV gene of bacteriophage T4 in Escherichia coli. AB - A 713-base-pair Hae III fragment from bacteriophage T4 encompassing the denV gene with its preceding promoter has been cloned in a pBR322-derived positive selection vector and introduced into a variety of DNA repair-deficient uvr and rec and uvr,rec Escherichia coli strains. The denV gene was found to be expressed, probably from its own promoter, causing pyrimidine dimer incision deficient uvrA, uvrB, uvrC strains to be rescued by the denV gene. A uvrD (DNA helicase II) strain was also complemented, but to a lesser extent. A wild-type strain did not seem to be affected at the UV doses tested. Surprisingly, all recA, recB, and recC strains tested also showed an increased UV resistance, perhaps by reinforcement of the intact uvr system in these strains. Complementation of denV- T4 strains and host-cell reactivation of lambda phage was also observed in denV+ E. coli strains. Equilibrium sedimentation showed that DNA repair synthesis occurred in a UV-irradiated uvrA E. coli strain carrying the cloned denV gene. Southern blotting confirmed our earlier results [Valerie, K., Henderson, E. E. & de Riel, J. K. (1984) Nucleic Acids Res. 12, 8085-8096] that the denV gene is located at 64 kilobases on the T4 map. Phage T2 (denV-) did not hybridize to a denV-specific probe. PMID- 2991892 TI - Application of the mini-Mu-phage for target-sequence-specific insertional mutagenesis of the herpes simplex virus genome. AB - An earlier technique for insertional mutagenesis of large viral genomes involved the insertion of the thymidine kinase (TK) gene at a specific target site, cotransfection of the fragment carrying the insertion with the intact viral genome, and selection of the progeny for viral recombinants expressing the TK gene. The inserted TK gene could then be replaced by cotransfection of the recombinant DNA with fragments carrying a foreign sequence or a deletion in the target sequence. To enable the probing of larger target domains and facilitate insertional mutagenesis, we extended this technique by insertion of a 2.2 kilobase-pair (kbp) herpes simplex virus 1 (HSV-1) chimeric alpha TK gene into the 7.5-kbp mini-Mu-phage (alpha TK-mini-Mu) and lysogenized Escherichia coli with the helper Mu phage and the alpha TK-mini-Mu. Induction of phage multiplication of the lysogenized E. coli after transformation with plasmids carrying HSV-1 DNA and subsequent infection of E. coli RecA+ lysogenized with Mu phage yielded plasmid populations carrying randomly inserted alpha TK-mini-Mu DNA. Application of this procedure for insertional mutagenesis of the BamHI B fragment, which spans the junction between the unique and reiterated sequences of the L component of viral DNA, yielded two types of recombinants. Viral recombinant designated RBMu1 contained the intact alpha TK-mini-Mu inserted into unique sequences of BamHI B fragment. In recombinant RBMu2, the alpha TK-mini-Mu was inserted at or in the repeated sequences, but approximately equal to 14 kbp comprising most of the internal reiterations of the viral genome in the prototype arrangement were deleted. PMID- 2991893 TI - Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion. AB - A method that allows the specific cloning of DNA fragments absent from patients homozygous or hemizygous for chromosomal deletions is described. The method involves phenol-accelerated competitive DNA reassociation and subsequent molecular cloning of appropriately reassociated molecules. The deletion DNA sample utilized in the competition was isolated from a patient with a minute interstitial deletion in the short arm of the X chromosome. Sheared DNA isolated from a male child, who was diagnosed as having Duchenne muscular dystrophy, chronic granulomatous disease, and retinitis pigmentosa, was combined in a 200 fold excess with Mbo I-cleaved DNA isolated from a 49, XXXXY human lymphoid cell line, and the mixture was subjected to a phenol-enhanced reassociation technique. Analysis of 81 unique segments derived from cloned reassociated DNA molecules has led to the identification of 4 (5%) human DNA fragments that are absent from the male patient's DNA. The 4 clones were localized, on the basis of hybridization with restriction nuclease-digested genomic DNA from a panel of human and human rodent hybrid cell lines, into three regions surrounding band 21 of the short arm of the normal human X chromosome. These clones are potential linkage markers for the diseases affecting this boy. Each clone, as well as others obtainable by this approach, may also serve as a starting point in the eventual cloning of these three X-linked-disease loci. Extension of this approach to other loci, including human tumors potentially homozygous for small deletions, should also be possible. PMID- 2991894 TI - Deletions within a defective suppressor-mutator element in maize affect the frequency and developmental timing of its excision from the bronze locus. AB - Six independent derivatives of the bz-m13 allele, which contains a 2.2-kilobase pair defective Suppressor-mutator (dSpm) insertion at the bronze (bz) locus, have been isolated and analyzed. The derivatives were selected for alterations in the frequency and timing of somatic reversion; such derivatives have previously been analyzed genetically and designated "changes in state" by McClintock [McClintock, B. (1955) Carnegie Inst. Washington, Yearb. 54, 245-255]. All of the derivatives analyzed in the present study revert substantially later in development than the original insertion mutation and some show a very low frequency of reversion as well. All of the derivatives contain insertions at the same site as the parent bz m13 allele. Deletions of 400-1300 base pairs were found in the dSpm elements in four of the six derivatives; the remaining derivatives could not be distinguished structurally from the original mutant allele. The results suggest that changes in the frequency and developmental timing of excision are attributable to alterations in the dSpm element. Furthermore, these data suggest that DNA sequences near the ends of the element are important for responding to the two transacting functions supplied by the transposition-competent Suppressor-mutator (Spm) element. PMID- 2991895 TI - Double recombination of a single immunoglobulin kappa-chain allele: implications for the mechanism of rearrangement. AB - DNA fragments containing immunoglobulin kappa-chain sequences from two different plasmacytomas (PC 3609 and PC 7043) were found by blot-hybridization studies to be dissociated from germ-line sequences on both the 3' and 5' ends. These fragments were cloned, sequenced, and found to contain the structural features of a product of two recombination events. Each contained a variable (V kappa) gene segment recombined with a joining (J kappa) gene segment followed by the characteristic kappa light chain V-J reciprocal structure, a 5' J kappa flanking sequence joined to a 3' V kappa flanking sequence. These segments of DNA represent double recombination products (DRPs) of the same kappa-chain allele. The DRP from PC 3609 contains a normal V-J1 recombination, while the DRP from PC 7043 contains an aberrant V-J2 recombination, resulting in a frameshift. The reciprocal structure in the PC 3609 DRP is the result of a V-J2 recombination; the reciprocal structure in the DRP of PC 7043 is the result of a V-J3 recombination and appears to have been derived directly from the productive kappa chain gene recombination in that plasmacytoma. These products demonstrate the capacity of a single kappa light chain immunoglobulin allele to undergo multiple V-J recombinations. Furthermore, the presence of a V-J recombination and its reciprocal product in the same cell is inconsistent with a segregating mechanism, such as sister chromatid exchange, but is consistent with an inversion mechanism. PMID- 2991896 TI - Genomic diversity of the acquired immune deficiency syndrome virus HTLV-III: different viruses exhibit greatest divergence in their envelope genes. AB - Converging lines of research have linked human T-cell lymphotropic virus type III (HTLV-III) to the pathogenesis of the acquired immune deficiency syndrome. A characteristic feature of this virus is its genomic heterogeneity, which occurs to varying degrees in different viral isolates. To define further the nature and extent of these genomic changes, we compared the molecularly cloned genomes of two variant HTLV-III isolates by extensive restriction enzyme mapping and heteroduplex thermal melt analysis. Both viral isolates were found to be highly related to each other throughout their entire genomic complement, yet they differed markedly in their restriction enzyme maps. Electron microscopic heteroduplex analysis revealed several distinct regions of divergence located almost exclusively in the part of the genome that encodes the viral envelope gene. In vitro culture of one of these viruses over a period of 3 months did not result in any genomic changes as determined by restriction analysis of viral DNA. These results, as well as the recently published nucleotide sequences of other HTLV-III isolates, indicate that the most substantial variation among HTLV-III isolates is located in the envelope. These findings raise the possibility that viral isolates from different individuals could have important biological differences in their envelope antigens, a consideration relevant to ongoing attempts to develop a vaccine against HTLV-III. PMID- 2991897 TI - Adenosine-activated potassium conductance in cultured striatal neurons. AB - We have examined the effect of adenosine on the membrane properties of cultured embryonic mouse striatal neurons using patch electrode techniques. Adenosine at 50 microM effectively blocked spontaneous action potential activity. Adenosine or 2-chloroadenosine caused a slow hyperpolarization of the membrane potential and, under voltage clamp, an outward current that was blocked by 1 mM theophylline. ATP also caused a hyperpolarization that was slower and weaker than the adenosine response and could be blocked by 1 mM theophylline. The current induced by adenosine appears to be carried by potassium since (i) an inward current was generated by adenosine when the cells were internally perfused with cesium salts and (ii) the reversal potential of the outward current shifted 57 mV with a 10 fold change in extracellular potassium concentration. The adenosine response is voltage dependent in that the current evoked by adenosine is reduced at holding potentials more positive than -55 mV, despite a larger driving force. Though calcium influx is not required for adenosine to activate the potassium conductance, some components of the cytosol may be essential, since the response is lost during intracellular perfusion. PMID- 2991898 TI - Transcription factor Sp1 recognizes promoter sequences from the monkey genome that are simian virus 40 promoter. AB - A 440-base-pair fragment of African green monkey genomic DNA shares homology with the transcriptional regulatory region of simian virus 40 (SV40) and has been reported to direct transcription in vivo. We find that two regions within this fragment bind the promoter-specific cellular transcription factor Sp1 and are protected in DNase protection ("footprinting") experiments. As in SV40, binding occurs in regions containing multiple copies of the sequence GGGCGG. These regions, when fused to the proximal, or "TATA box," element of the herpes simplex virus thymidine kinase promoter, are able to direct Sp1-dependent transcription in vitro. The finding that Sp1 is capable of productive interaction with sequences taken from a cellular promoter supports the idea that Sp1 may play a role in modulating transcription of cellular genes. PMID- 2991900 TI - A glycoprotein associated with the non-A, non-B hepatitis agent(s): isolation and immunoreactivity. AB - A glycoprotein was isolated and purified to homogeneity from the serum of a patient with chronic non-A, non-B hepatitis. NaDodSO4/PAGE of the glycoprotein revealed a single major band at Mr approximately 77,000. Antibodies to this glycoprotein were shown to possess the following immunoreactivity: (i) they reacted by radioimmunoassay with sera obtained at the time of diagnosis from 17 of 42 patients with non-A, non-B hepatitis and with only 2 of 58 sera from either matched controls or patients with hepatitis A or hepatitis B, (ii) they reacted with sucrose gradient fractions from a proven infectious non-A, non-B hepatitis serum at a peak density of 1.14 g/ml and in the soluble protein fractions on top of the gradient, and (iii) they reacted in ELISA with disrupted human T-cell lymphocytotropic virus type III (HTLV-III), and (iv) they reacted in immunoblots with a protein of Mr 74,000 derived from HTLV-III. PMID- 2991899 TI - Characterization and sequence of the promoter region of the human epidermal growth factor receptor gene. AB - The promoter region of the epidermal growth factor (EGF) receptor has been identified by in vitro transcription using EGF receptor genomic DNA fragments as template and by primer extension and nuclease S1 mapping using EGF receptor mRNA. Six transcriptional start sites were identified. DNA sequence analysis shows that the promoter region contains neither a "TATA box" nor a "CAAT box," has an extremely high G+C content (88%), and contains five CCGCCC repeats and four (TCC)TCCTCCTCC repeats. This promoter region is situated close to or within a DNase I-hypersensitive site in A431 human epidermoid carcinoma cells, which overproduce the EGF receptor. The EGF receptor gene promoter has some resemblance to the promoter of the hydroxymethylglutaryl-CoA reductase gene and the early promoter of simian virus 40. This similarity may offer a clue to the mechanism by which the receptor gene is regulated. PMID- 2991901 TI - Differentiation-enhanced binding of the amino-terminal fragment of human urokinase plasminogen activator to a specific receptor on U937 monocytes. AB - The purified amino-terminal fragment (ATF) of human urokinase plasminogen activator (residues 1-135), which is not required for activation of plasminogen, binds with high affinity to specific plasma membrane receptors on U937 monocytes. Intact urokinase efficiently competes for 125I-labeled ATF binding; 50% competition occurs with 1 nM urokinase. A large part of receptor-bound urokinase remains on the cell surface for at least 2 hr at 37 degrees C. Differentiation of U937 monocytes into macrophage-like cells specifically increases ATF binding 10- to 20-fold. These results suggest an important role for urokinase in monocyte/macrophage biology: the native enzyme binds to the cells with the amino terminal domain; the catalytic, carboxyl-terminal domain remains exposed on the cell surface to stimulate localized proteolysis and facilitate cell migration. PMID- 2991902 TI - Purification of the opiate receptor of NG108-15 neuroblastoma-glioma hybrid cells. AB - Opiate receptors from NG108-15 neuroblastoma-glioma hybrid cell membranes were purified to apparent homogeneity in a form covalently labeled with the opiate affinity ligand 3-methylfentanylisothiocyanate (super-FIT). The purification procedure consists of five steps: quantitative labeling of opiate receptors in membranes with super-FIT; solubilization in a lubrol/3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate mixture; adsorption to and elution from a wheat germ agglutinin-Sepharose column; immunoaffinity chromatography on columns of immobilized anti-fentanyl; preparative gel electrophoresis in the presence of NaDodSO4. The protein was purified approximately 30,000-fold in 3% overall yield to a state in which there is 1 mol of super-FIT bound per mol of protein, corresponding to 21,000 pmol per mg of protein, the theoretically expected specific activity. The protein is glycosylated and migrates on NaDodSO4 gel electrophoresis with a Mr near 58,000. It has a strong tendency to dimerize, even in the presence of denaturing detergents, and it exists primarily as an oligomer in nondenaturing detergents. PMID- 2991903 TI - Removal of a 67-base-pair sequence in the noncoding region of protooncogene fos converts it to a transforming gene. AB - Transformation of fibroblasts by protooncogene fos (c-fos) requires the linkage of viral long terminal repeat (LTR) sequences and interruption of 3'-noncoding sequences. We have identified an A + T-rich stretch of 67 nucleotides, located 627-693 base pairs downstream from the coding domain and 123-189 base pairs upstream from the putative poly(A) addition site, removal of which confers transforming activity to the c-fos gene. A novel regulation of the expression of the c-fos gene is proposed, which may be functional in vivo to prevent the gene from becoming an oncogene. PMID- 2991904 TI - Ectopic peptides released by a human melanoma cell line that modulate the transformed phenotype. AB - Cells derived from a human melanoma strain by low-serum selection in monolayer were found to be capable of growth in semisolid medium, forming colonies ranging from tight to loose or dispersed. These phenotypes were found to be stable after cloning and retesting. Examination of the biological activities of ectopic peptides produced by a clone that gives rise to loose dispersed colonies revealed the presence of a transforming growth factor beta (TGF-beta; apparent Mr 14,700) and a single TGF-alpha species produced at an unusually high concentration (2.2 ng/ml). Coeluted with the TGF-alpha at approximately 22,500 was a mitogenic activity and a previously undescribed activity capable of modulating the phenotype of induced normal rat kidney fibroblast (NRK-49F) colonies in semisolid medium. This activity can modulate the usual tight colony to express a loose or dispersed phenotype characteristic of the producer cells. Both the possible role these ectopic peptides play in the expression of the transformed phenotype by the tumor cells producing them and the possible correlation between the Mr 22,500 epidermal growth factor-like peptide released by this particular tumor line and high molecular weight epidermal growth factor-like peptides found in the urine of cancer patients are discussed. PMID- 2991905 TI - Purification and characterization of caldesmon77: a calmodulin-binding protein that interacts with actin filaments from bovine adrenal medulla. AB - Caldesmon150, a protein composed of the Mr 150,000/147,000 doublet, alternately binds to calmodulin and actin filaments in a Ca2+-dependent "flip-flop" fashion. In all fibroblast cell lines examined, we also found a Mr 77,000 protein that crossreacts with anti-caldesmon150 antibody by using an immunoprecipitation technique [Owada, M.K., Hakura, A., Iida, K., Yahara, I., Sobue, K. & Kakiuchi, S. (1984) Proc. Natl. Acad. Sci. USA 81, 3133-3137]. In this report, we examine the tissue distribution of caldesmon by the method of immunoblotting, using caldesmon-specific antibody. Both caldesmon150 and caldesmon77 show widespread distribution in the tissues examined. Caldesmon77 is more widely distributed than caldesmon150, and we have purified caldesmon77 from bovine adrenal medulla. Its molecular weight estimated by NaDodSO4/polyacrylamide gel electrophoresis was 77,000, and a tetramer of this polypeptide may constitute the native molecule (Mr, 300,000). Caldesmon77 possesses a number of features in common with caldesmon150, including flip-flop binding to calmodulin and actin filaments depending on the concentration of Ca2+ and crossreactivity with caldesmon150 specific antibody. Analysis of caldesmon77-F actin interaction by sedimentation and electrophoresis revealed that 0.5 mg of caldesmon77 bound to 1 mg of F actin. This indicated that the molar ratio between caldesmon77 (tetramer) and actin monomer was calculated to be 1:12-14. In addition, caldesmon77 regulated the actin-myosin interaction in Ca2+-sensitive actomyosin obtained from adrenal medulla. These results suggest that caldesmon77 might be a ubiquitous actin linked regulator of nonmuscle contractile processes, including those in adrenal medulla. PMID- 2991906 TI - A stochastic mechanism controls the relative replication of equally competent ribosomal RNA gene sets in individual dipteran polyploid nuclei. AB - The endoreplication of the two nucleolar organizers (NOs) of the diploid genome has been examined in individual polyploid nuclei of the dipteran Calliphora erythrocephala. Crosses between two strains with diagnostic nontranscribed spacer polymorphisms in their rRNA genes were used to provide progeny with distinguishable NOs, and single nuclei of two highly polyploid cell types- salivary gland and nurse cells--were examined from individual F1 animals. Initially the representation of the two NOs in total polyploid tissue DNA was determined. This revealed that, although the NO regions present in one of the strains (Tom) were very similar in spacer composition, they displayed two types of behavior in the hybrids containing the single NO region typical of the second strain (Karla). In TW phenotype F1 progeny, very little replication of the Tom NO relative to the Karla NO occurred, whereas in TS phenotype progeny replication of the Tom and Karla NOs was approximately equivalent. When individual polyploid nuclei of the TS phenotype animals were examined, however, the relative replication of the Tom and Karla NOs was found not to be a fixed genetic property but to vary dramatically from cell to cell. This was true even for the nurse cell nuclei within a single ovarian follicle, which are the products of only four mitotic divisions of a single germ-line cell. These findings indicate that for NOs of similar replicative competence, a stochastic mechanism governs the relative usage of each NO for endoreplication and that the relative activity of the two NOs is not stably determined through the mitotic divisions preceding polyploidization. Stochastic selection after mitotic DNA replication could be a general phenomenon governing the relative usage (transcription) of different, but equally competent, alleles of any gene in individual cells, if the required factors are in short supply. PMID- 2991907 TI - Isolation of the gene encoding adenylate cyclase in Saccharomyces cerevisiae. AB - By complementation of the cyr1-1 mutation in Saccharomyces cerevisiae, we have isolated yeast genomic DNA containing the structural gene that encodes the catalytic unit of adenylate cyclase (EC 4.6.1.1). The isolated DNA restored adenylate cyclase activity to cyr1-1 mutants and directed integration at the CYR1 locus. Wild-type strains transformed with CYR1 DNA on the high copy number vector YEp24 contained 4- to 6-fold more adenylate cyclase activity than strains carrying the plasmid with no insert. This result suggests that expression of the CYR1 gene product, rather than that of other polypeptide components of the adenylate cyclase system, limits total adenylate cyclase activity in S. cerevisiae. CYR1-containing plasmids also complemented the temperature-sensitive growth defect of the cell division cycle mutation cdc35-1, which confers a phenotype under restrictive conditions similar to that of cyr1-1 and maps to the same locus. Further, cdc35-1 cam mutants, which contain mutations that enable them to take up cAMP from the medium, grew at the restrictive temperature in the presence of exogenous cAMP. These observations support the view that CDC35 and CYR1 are allelic and confirm the hypothesis that cAMP synthesis is required for cells to pass through the "start" position of the cell division cycle. PMID- 2991908 TI - Linkage map of the short arm of human chromosome 11: location of the genes for catalase, calcitonin, and insulin-like growth factor II. AB - The following order of genes on the short arm of human chromosome 11 (11p) was determined previously: parathyroid hormone (PTH)-the beta-globin gene cluster (HBBC)-HRAS1/insulin. Although it is generally agreed that HRAS1 (formerly termed c-Ha-ras-1) and the insulin gene are close to each other [1-4 centimorgans (cM)], their order on chromosome 11p is still in question. We have now added three other genes, those for catalase, calcitonin, and insulin-like growth factor II (IGF II), to this map of chromosome 11p by use of restriction site polymorphisms adjacent to these genes in classical linkage analysis. Most importantly, we find no evidence of linkage between the catalase and HBBC loci. In addition, our data indicate that the calcitonin gene is located between the catalase gene and the PTH gene. Our best estimate of the distance between the catalase and calcitonin gene is approximately 16 cM, while that between the calcitonin and PTH genes is approximately equal to 8 cM. In agreement, very loose linkage was found between the catalase and PTH loci (approximately 26 cM). Since the catalase locus has been mapped to 11p13, these data support the view that the PTH, HBBC, HRAS1, and insulin loci are located on the distal short arm of chromosome 11. The IGF-II gene is tightly linked to both the HRAS1 oncogene and the insulin gene since no recombinants were observed between the IGF-II and the HRAS1/insulin loci. Thus, based on our linkage analysis we propose that the most likely gene order for the short arm of chromosome 11 is centromere-catalase-calcitonin-PTH-HBBC HRAS1/insulin-tel ome re and that the IGF-II gene is very close to both the HRAS1 and the insulin genes. PMID- 2991909 TI - Hemin inhibits internalization of transferrin by reticulocytes and promotes phosphorylation of the membrane transferrin receptor. AB - Addition of hemin to reticulocytes inhibits incorporation of iron from transferrin [Ponka, P. & Neuwirt, J. (1969) Blood 33, 609-707]. Heme also regulates protein synthesis in immature erythroid cells through its effects on phosphorylation of the initiation factor eIF-2. We have therefore examined its effects on endocytosis of iron-transferrin and phosphorylation of the transferrin receptor. Hemin (10-50 microM) reduced iron transport but increased cell associated transferrin. When intracellular iron delivery was inhibited by NH4Cl, no such increase in cell-associated transferrin was seen. During uptake of 125I labeled transferrin in the steady state, the use of a washing technique to dissociate bound transferrin on the cell membrane showed that radioligand accumulated on the surface of hemin-treated cells. Hemin reduced the initial influx of transferrin, thereby diminishing incorporation of iron. Receptor phosphorylation was investigated by immunoprecipitation of reticulocyte extracts after metabolic labeling with [32P]Pi. In the absence of ligand, phosphorylated receptor was chiefly localized on cell stroma. Exposure to transferrin increased cytosolic phosphorylated receptor from 15-30% to approximately 50% of the total, an effect overcome by hemin treatment. Addition of hemin in the presence of transferrin enhanced net phosphorylated receptor in the reticulocyte in association with a redistribution of phosphorylated receptor to stromal membranes. The findings suggest a possible relationship of phosphorylation to endocytosis of the transferrin receptor in reticulocytes. PMID- 2991910 TI - Oxidant-dependent metabolic activation of polycyclic aromatic hydrocarbons by phorbol ester-stimulated human polymorphonuclear leukocytes: possible link between inflammation and cancer. AB - Oxidants, such as those generated by metabolically activated phagocytes in inflammation, have been implicated in the metabolic activation of carcinogens, and in this study we demonstrate that the interaction of (+/-)-trans-7,8 dihydroxy-7,8-dihydrobenzo[a]pyrene (BP 7,8-dihydrodiol) with phorbol ester stimulated polymorphonuclear leukocytes (PMNs) results in the generation of both a chemiluminescent intermediate and one that covalently binds to DNA. Cu(II)(3,5 diisopropylsalicylic acid)2 (CuDIPS), a biomimetic superoxide dismutase, and azide, a myeloperoxidase inhibitor, inhibited both of these reactions, indicating a dependency on oxygen-derived oxidants in these hydrocarbon-activation processes. Concordant with the formation of a carcinogen-DNA adduct, the admixture of BP 7,8-dihydrodiol and phorbol ester-stimulated PMNs elicited mutagenesis in Salmonella typhimurium strain TA100. 7,8-Dihydro-BP and BP cis-7,8 dihydrodiol were also mutagenic, whereas derivatives lacking a double bond at the 9,10 position were not. These results demonstrate that oxidants generated by metabolically stimulated PMNs can activate penultimate polycyclic aromatic hydrocarbons to a genotoxic metabolite and further defines a role for inflammation in carcinogenesis. PMID- 2991911 TI - Monoclonal antibodies specific for p24, the major core protein of human T-cell leukemia virus type III. AB - Four mouse hybridomas secreting monoclonal antibodies specific for p24, the major core antigen of the human T-cell leukemia virus type III (HTLV-III), have been developed, and their specificities have been partially characterized. These antibodies specifically recognized p24 of HTLV-III in extracts of HTLV-III and in HTLV-III-producing cells. No epitopes cross-reactive with HTLV-I and -II were detected with these antibodies. These hybridomas will be extremely valuable reagents in identifying expression of HTLV-III in infected cultures and in cells or tissues from patients with suspected immunodeficiency syndrome. PMID- 2991912 TI - Peripheral-type benzodiazepines influence ornithine decarboxylase levels and neurite outgrowth in PC12 cells. AB - A number of the benzodiazepines (BZDs) inhibit nerve growth factor (NGF)-induced neurite outgrowth in a dose-dependent manner in PC12 cell cultures. The rank order of potency of a series of BZDs for inhibition of neurite outgrowth does not correlate with the order of their affinity constants for the so-called peripheral BZD sites present on PC12 cells. Whereas the inhibition of neurite extension is stereospecific, the binding to the peripheral site is not. The inhibition of neurite outgrowth is not attributable to a blockade of NGF binding to either its fast or slow receptors. Additionally, several other characteristics of NGF stimulation of PC12 cells, such as autoadhesion and the induction of ornithine decarboxylase (OrnDCase), remain unaltered in the presence of BZDs. Many BZDs increase OrnDCase levels in PC12 cells in the absence of NGF. The OrnDCase response to BZDs is blocked by actinomycin D. Furthermore, the structural requirements of BZDs for induction of OrnDCase activity is not identical to that for the inhibition of NGF-induced neurite extension. Thus, unlike attenuation of neurite extension, BZD induction of OrnDCase is not stereospecific. Also, some BZDs block neurite growth but do not induce OrnDCase. We propose that there are at least three sites of action of BZDs on PC12 cells. The first is the well characterized high-affinity peripheral site. The second distinct locus of action results in a blockade of NGF-induced neurite outgrowth. The structural requirements for the latter effect are essentially indistinguishable from those for BZD induction of hemoglobin synthesis in Friend erythroleukemia cells. The third site of action results in an induction of OrnDCase. Structural requirements for this activity are not identical to those for either the differentiation of Friend cells or the inhibition of neurite outgrowth. PMID- 2991913 TI - Inositol trisphosphate-induced calcium release and contraction in vascular smooth muscle. AB - Inositol 1,4,5-trisphosphate (InsP3) caused Ca release and tension development in rabbit main pulmonary artery smooth muscle permeabilized with saponin or digitonin. Both of these responses to single additions of InsP3 (0.5-30 microM) were repeatable and occurred in the presence of 0.0-1.9 mM free Mg2+. Sustained contractions were induced by InsP3. The amount of Ca released by InsP3, measured with a Ca2+-selective electrode, was also estimated to be sufficient to stimulate contraction in intact smooth muscle. Ca release was not influenced by inhibitors of mitochondrial oxidative phosphorylation. The uptake of Ca2+ from the medium into the InsP3-sensitive pool was ATP-dependent. The present results support the hypothesis that, in smooth muscle, InsP3 is the messenger, or one of the messengers, involved in transmitter-induced (pharmacomechanical) Ca release from the sarcoplasmic reticulum, which is the intracellular Ca store identified previously as the source of Ca released by norepinephrine in main pulmonary artery. PMID- 2991914 TI - Identification of a nucleic acid helix-destabilizing protein from rat liver as lactate dehydrogenase-5. AB - A rat liver DNA helix-destabilizing protein (HDP) that has previously been proposed to play a role in transcription has been identified as M chain lactate dehydrogenase (LDH-5; L-lactate:NAD+ oxidoreductase, EC 1.1.1.27). Tryptic peptides accounting for 157 amino acids in the rat liver HDP have been characterized and then matched to the published sequence for the M chain of porcine LDH. Based on amino acid compositions and direct solid-phase protein sequencing, at least 148 of the 157 residues that were compared are identical in both proteins. In addition, both porcine LDH and the rat liver HDP have blocked amino termini and similar amino acid compositions and molecular weights. Rat liver HDP and LDH-5 that were purified to molecular homogeneity had similar specific activities in both single-stranded DNA (ss DNA) binding and LDH assays. HPLC tryptic peptide maps were also identical for both the rat liver HDP and LDH proteins. Since preincubation of HDP in NADH prevents its binding to ss DNA, both NADH and ss DNA may be binding at the same site. Further support for this latter idea derives from chemical-modification studies which demonstrate that tyrosine 238, which is located near the coenzyme binding site of LDH, seems to be essential for the ability of HDP to bind ss DNA. These results indicate caution in ascribing in vivo roles solely on the basis of binding to ss DNA. Alternatively, they suggest that a single protein may play more than one biological role. PMID- 2991915 TI - Identification of immediate early genes from herpes simplex virus that transactivate the virus thymidine kinase gene. AB - A HeLa cell transient-expression assay system was used to determine if isolated immediate early (alpha) genes from herpes simplex virus (HSV) could transcriptionally activate (transactivate) the type 1 (HSV-1) thymidine kinase (TK) gene [an early (beta) gene]. Cells transfected with the TK gene alone transcribed very low levels of TK RNA. Cells cotransfected with plasmids bearing the sequences that encode the alpha-gene product infected cell protein 0 or 4 (ICP0 or ICP4) and the TK gene faithfully transcribed high levels of TK RNA. The plasmid containing the sequences encoding ICP0 was a more potent transactivator than the plasmid containing the sequences for ICP4. PMID- 2991916 TI - In vitro mutagenesis of the v-sis transforming gene defines functional domains of its growth factor-related product. AB - The polypeptide sequence of the v-sis transforming gene product of simian sarcoma virus (SSV) can be divided into four regions that are likely to represent structural domains of the protein. Mutations were generated in the SSV nucleotide sequence to assay the extent or function of each of these regions. The results indicate that the helper virus-derived amino-terminal sequence as well as a core region homologous to polypeptide chain 2 of platelet-derived growth factor (PDGF) are required for the transforming function of the protein. Products of transforming but not nontransforming mutants formed dimer structures conformationally analogous to biologically active PDGF. PMID- 2991917 TI - Cloning and characterization of cDNAs encoding 2,3,7,8-tetrachlorodibenzo-p dioxin-inducible rabbit mRNAs for cytochrome P-450 isozymes 4 and 6. AB - Monoclonal antibodies toward rabbit liver cytochrome P-450 isozyme 6 (P-450 6) were used to identify several recombinant clones from a pBR322 cDNA library that express beta-lactamase-P-450 6 hybrid proteins. The nucleic acid sequence and predicted amino acid sequence of a rabbit P-450 6 cDNA shows a high degree of homology with rat P-450c and mouse P1-450. When used as a probe to rescreen the library, the P-450 6 cDNA hybridized to several heterologous classes of cDNAs. One such class was shown to encode P-450 4 by comparison of its predicted amino acid sequence to amino acid sequences of cysteine-containing tryptic peptides derived from P-450 4. DNA sequence analysis of a cDNA clone belonging to a third class demonstrated that it contained a 131-base-pair intervening sequencing when compared to the cDNA coding for P-450 6. This sequence corresponds in location to intron E of the rat P-450c gene. Blot-hybridization analysis demonstrated that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) dramatically induced P-450 4 and 6 mRNAs, which differ in size. The sizes of these mRNAs differ from their analogs in the mouse as a result of divergence in the 3' untranslated portions of the mRNAs. PMID- 2991918 TI - Structural similarities between the mammalian beta-adrenergic and reovirus type 3 receptors. AB - The mechanism by which viruses bind to and infect specific tissues to cause disease has only recently begun to be understood. The mammalian reoviruses provide an especially attractive model for studying the details of cell surface recognition. The cell and tissue tropism of reovirus is determined by a portion of the viral hemagglutinin termed the neutralization domain. We have reported previously on the generation of both monoclonal and polyclonal anti-idiotypic antibodies that mimic the viral hemagglutinin in the specificity of binding to the reovirus receptor. By using these anti-idiotypic antibodies as specific probes, we have successfully isolated the mammalian reovirus receptor from neuronal and lymphoid cells. In the present study, we report that the reovirus receptor is structurally similar to the mammalian beta-adrenergic receptor. This conclusion is based on the following observations: (i) purified beta-adrenergic receptor is immunoprecipitable by anti-reovirus receptor antibody; (ii) purified reovirus receptor obtained from murine thymoma cells and beta-adrenergic receptor obtained from calf lung exhibit identical molecular masses and isoelectric points; (iii) trypsin digests of purified reovirus and beta-adrenergic receptors display indistinguishable fragment patterns; (iv) purified reovirus receptor binds the beta-antagonist [125I]iodohydroxybenzylpindolol and this binding is blocked by the beta-agonist isoproterenol. PMID- 2991919 TI - Potential role of the src gene product in inhibition of gap-junctional communication in NIH/3T3 cells. AB - The effects of the src gene on the activity of protein kinase C and intercellular communication have been studied in transformed NIH/3T3 clones isolated from soft agar following transfection with the plasmid carrying the v-src gene (psrc-11). Six transformed clones that were studied contained newly incorporated v-src genes in the genome, had an increased amount of pp60src, and showed enhanced activities of protein kinase C. Intercellular communication, studied by observing with autoradiography the transfer of [3H]uridine nucleotide from prelabeled donor cells to recipient cells in contact, was found to be reduced in transformed clones as compared to parental NIH/3T3 cells. Treatment with phorbol 12-myristate 13-acetate was also found to increase protein kinase C activity and to reduce intercellular communication in normal NIH/3T3 cells. These results suggest that the v-src gene product, in a manner similar to some of the powerful tumor promoters, may directly or indirectly affect cell-cell communication. PMID- 2991920 TI - Differentiation of a human leukemia cell line and expression of collagenase inhibitor. AB - A human collagenase inhibitor (CI) of Mr 28,500 has been extensively characterized in skin fibroblasts and identified in a variety of connective tissues. Because human alveolar macrophages synthesize and secrete both a collagenase and CI that are immunologically and functionally identical to their counterparts in fibroblasts, we studied the production of such proteins by an immature human cell line (HL60) that can be induced to differentiate along monocytic or granulocytic pathways. The cells failed to synthesize collagenase under any culture condition tested. However, upon exposure to 1,25 dihydroxyvitamin D3 or phorbol esters (PMA), both of which promote monocytic differentiation of HL60, these cells synthesized and released CI in a dose dependent manner. Furthermore, the extent of CI expression was paralleled by the acquisition by such cells of the monocytic marker 63D3, indicating that inhibitor production and differentiation are closely correlated. This CI was immunologically and functionally identical to that produced by human macrophages and human skin fibroblasts. The quantity of CI synthesized by PMA-stimulated cells was 3- to 5-fold greater than produced by human alveolar macrophages, approximately equal to 1 microgram per 10(6) cells per day. In contrast, undifferentiated HL60 cells produced little or no detectable CI (less than or equal to 10-20 ng per 10(6) cells per day). Interestingly, when HL60 cells were stimulated to undergo granulocytic differentiation by dimethyl sulfoxide or retinoic acid, they also produced the "monocytic" CI. PMID- 2991921 TI - Cultured psoriatic fibroblasts from involved and uninvolved sites have a partial but not absolute resistance to the proliferation-inhibition activity of 1,25 dihydroxyvitamin D3. AB - We examined the responsiveness of cultured dermal fibroblasts from biopsies of uninvolved and involved areas of skin from six patients with psoriasis to the cell-proliferation-inhibition activity of 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D3). Cultured fibroblasts from age-matched controls responded to 1,25-(OH)2-D3 (at 0.01, 1, 10, and 100 microM) in a dose-dependent fashion, whereas cultured psoriatic fibroblasts from involved or uninvolved skin showed no inhibition of proliferation when exposed to 0.01 or 1 microM of 1,25-(OH)2-D3. However, 1,25 (OH)2-D3 did inhibit proliferation of cultured psoriatic fibroblasts when the concentrations were increased to 10 and 100 microM. An analysis of the 1,25-(OH)2 D3 receptors in cultured psoriatic fibroblasts from uninvolved skin revealed that the Kd, nmax, and sedimentation coefficient were identical to the receptors found in the fibroblasts from age-matched controls. Therefore, cultured psoriatic fibroblasts from involved and uninvolved skin have a partial resistance to 1,25 (OH)2-D3, suggesting that there may be a biochemical defect that is inherent in the dermal fibroblasts of psoriatic patients. Recognition of this defect may provide a new approach for the evaluation of the cause and treatment of this disfiguring skin disorder. PMID- 2991922 TI - Activation regions in a yeast transposon have homology to mating type control sequences and to mammalian enhancers. AB - The DNA sequence of the Ty1 activating region from the CYC7-H2 mutant of Saccharomyces cerevisiae is presented. Analysis of the data revealed the presence of four simian virus 40-type enhancer core sequences. Two of the Ty1 enhancer cores are contiguous with sequences also homologous to the diploid control site at MAT alpha. We postulate that these two Ty1 regions of approximately equal to 30 base pairs are regulatory blocks, and we have analyzed deletions to ascertain whether they are necessary for effects of Ty1 on adjacent gene expression. We found that activation is lost when a restriction fragment encompassing both postulated regulatory blocks is deleted. Deletion of restriction fragments that remove only one of the two regulatory blocks has little or no effect on Ty1 activating ability in haploid yeast cells or on repression of this function in diploid yeast cells. Because the most significant internal homologies in the restriction fragments analyzed are the putative regulatory blocks, these observations suggest that enhancer-like sequences are involved in cell-type control of Ty1 effects on gene expression. PMID- 2991924 TI - Exuberant restriction fragment length polymorphism associated with the DQ alpha chain gene and the DX alpha-chain gene. AB - Cellular DNAs from individuals of 23 families were digested with five restriction endonucleases (Pvu II, EcoRI, HindIII, BamHI, and EcoRV) and then probed with a DX alpha-chain gene probe. Seventeen allogenotopes were observed, each of which could be assigned to a serologically defined haplotype by noting its segregation in families. Six sets of allogenotopes forming allelic series were noted. In comparison with restriction maps of the DQ alpha and the DX alpha regions, each of these series has been assigned to the DQ alpha or the DX alpha locus. Allogenotopes of the four DQ alpha series constitute three clusters correlating with the supertypic groups of class II histocompatibility antigens DQw1 (DR1, DR2, and DRw6), DRw53 (DR4, DR7, and DR9), and DR3 plus DR5 plus DR8. These 13 DQ alpha fragments constitute 22 different patterns. The two DX alpha series constitute two clusters, one of which is not found to be correlated strongly with DR specificities, whereas the other is correlated loosely (r = 0.45) with DR5 and DR7. This absence of strong linkage disequilibrium between the DX alpha series and the DR series contrasts with the DQ alpha series and suggests a recombination point between DQ alpha and DX alpha loci. PMID- 2991923 TI - Transposable element sequences involved in the enhancement of yeast gene expression. AB - The his4-917 mutation of yeast results from the insertion of a Ty element, Ty917, into the 5' regulatory region of the HIS4 gene. Ty917 prevents HIS4 transcription, thus rendering the cell histidine requiring. Recombination between Ty917 and a Ty element elsewhere in the yeast genome can result in the replacement of part or all of the Ty917 element by sequences from the Ty element. Recombinant derivatives display a variety of phenotypes including His-, weakly His+, and strongly His+. In most of the His+ derivatives, the expression of HIS4 is controlled by genes at the mating type locus. To identify the Ty sequences important in controlling the expression of an adjacent gene, we used Ty elements that have different effects on gene expression to construct hybrid Ty elements in vitro. The effects of these hybrid elements on HIS4 expression were examined. These experiments indicate that the critical sequence differences between Ty elements that permit HIS4 expression and those that prevent its expression lie in the rightmost (HIS4-proximal) 730 base pairs of the element. The DNA sequence of this region was determined for three elements: Ty917, which prevents HIS4 expression; Ty917(467), which confers a weak His+ phenotype; and Ty917(480), which confers a strong His+ phenotype. Within this region, Ty917(467) differs from Ty917 by a single base-pair change that is in the internal (epsilon) region of the Ty element. Ty917(480) differs from Ty917 by this same base-pair change and by 10 changes in the terminal delta sequence. The sequence change common to Ty917(467) and Ty917(480) lies in a region of the Ty element that is homologous to the simian virus 40 enhancer of transcription. PMID- 2991925 TI - The Escherichia coli supX locus is topA, the structural gene for DNA topoisomerase I. AB - Mutations in the supX locus, which result in the absence of DNA topoisomerase I enzyme activity in both Salmonella typhimurium and Escherichia coli, are all selected as suppressors of the leu-500 promoter mutation in S. typhimurium. To determine whether the supX locus is the structural gene topA for the DNA topoisomerase I enzyme or is a positive-acting regulator/activator gene for a nearby topA structural gene, nonsense mutations were selected in the E. coli supX gene carried on an F' episome in S. typhimurium cells. The cysB-topA region of the episomes with nonsense-mutant supX alleles were then cloned onto plasmid pBR322 and transformed into E. coli cells lacking a chromosomal supX gene. Three such E. coli strains, each carrying cloned DNA from episomes with different nonsense-mutant supX alleles, all lacked DNA topoisomerase I activity but expressed antigenic determinants specific to the enzyme; control cells lacked both enzyme activity and antigenic determinants. Maxicell studies of plasmid coded proteins demonstrated the absence of the DNA topoisomerase I protein (100 kDa) in the three strains but the appearance of a new smaller peptide in each (36, 47, and 64 kDa). These new peptides must represent fragments of the enzyme resulting from translation termination at the supX nonsense codons and confirm the interpretation that the supX gene is topA, the structural gene for DNA topoisomerase I. PMID- 2991926 TI - Heterogeneity in type I Gaucher disease demonstrated by restriction mapping of the gene. AB - A cloned fragment of human glucocerebrosidase cDNA has been used as a probe to study restriction polymorphisms in the region of the gene for Gaucher disease. Variability in the size of fragments produced by digestion with the restriction endonucleases Pvu II and Kpn I was discovered. The Pvu II polymorphism was found to be a very prevalent one with a gene frequency of 0.65 for the Pv1.1- allele and 0.35 for the Pv1.1+ allele. Similar frequencies were encountered among diverse ethnic groups. Five of eight Jewish patients with Gaucher disease were found to be heterozygous for the Pvu II restriction polymorphism. One non-Jewish patient with type I Gaucher disease was heterozygous for the Kpn I variant. The existence of Gaucher disease genes in association with either allele of the ancient Pvu II polymorphism clearly indicates that, even within the Jewish population, the Gaucher disease mutation has occurred independently more than once. Presumably, different mutations have also occurred in the non-Jewish population. PMID- 2991927 TI - Accurate transcription of cloned Neurospora RNA polymerase II-dependent genes in vitro by homologous soluble extracts. AB - We have developed soluble extracts from Neurospora crassa capable of accurately initiating the transcription of cloned Neurospora protein-encoding genes by RNA polymerase II in vitro. The genes encoding glutamate dehydrogenase (am) and histones H3 and H4 were transcribed by the extracts, and transcription was sensitive to alpha-amanitin at 1 mg/ml. The 5' heterogeneity of the in vitro initiation reactions was highly specific. Of the 17 transcription initiation sites within the inducible qa gene cluster, only one minor site was used in vitro, suggesting that, in general, transcription from qa gene promoters requires at least one different protein from those required for transcription of the am and histone genes. PMID- 2991928 TI - Nonrandom association of a type II procollagen genotype with achondroplasia. AB - Achondroplasia is an autosomal dominant disorder that involves defective endochondral bone formation. Type II collagen is the predominant collagen of cartilage. We found a HindIII polymorphic site in the normal Caucasian population by using the type II procollagen gene probe pgHCol(II)A. The presence of this site yields a 7.0-kilobase (kb) band; its absence yields a 14.0-kb band. We found a significant deviation in genotype distribution and allele frequencies in a population of unrelated individuals with sporadic achondroplasia, compared with the normal control population. The HindIII genotype frequencies in 32 individuals with achondroplasia are 0.41 for the 7/7 genotype (controls, 0.08), 0.34 for the 7/14 genotype (controls, 0.54), and 0.25 for the 14/14 genotype (controls, 0.37). The apparent equilibrium excess of the "7" allele in individuals with achondroplasia may reflect either a predisposition for the mutation that causes achondroplasia or it could be the result of the achondroplasia-causing mutation. In either case, these findings suggest an association of the type II procollagen gene with achondroplasia. PMID- 2991929 TI - Immunoglobulin gene rearrangements and deletions in human Epstein-Barr virus transformed cell lines producing different IgG and IgA subclasses. AB - During differentiation B lymphocytes may switch from the expression of surface IgM to the synthesis of IgG, IgA, or IgE isotypes by using a different heavy chain constant region (CH) gene. The molecular mechanisms by which switching occurs remain controversial. Rearrangements and deletions of CH genes 5' to the expressed gene have often been observed in the mouse and, more recently, in human cells that have switched isotypes. We have used human JH, C micro, C gamma, and C alpha probes to examine the extent of the deletions and rearrangements in clones of Epstein-Barr virus-transformed human cells that produce IgG1, IgG3, IgG4, or IgA1. Though deletions of CH genes 5' to the expressed CH gene were consistently observed, the rearrangement process appeared to be highly variable for the nonproductive CH gene locus: deletion or persistence of 5' CH genes, combinations of deletion and duplication of 5' genes, and deletions extending to 3' CH genes. Our results reveal an unexpected lack of specificity in the DNA deletions in cells that have undergone isotype switching. PMID- 2991930 TI - Expression in L cells of transfected class I genes from the mouse major histocompatibility complex. AB - One of the major surprises of the molecular analysis of major histocompatibility complex (MHC) genes is the large number of class I (K/D)-related sequences in the genome. Both restriction fragment length polymorphisms and cosmid cloning experiments showed them all to be closely linked to the MHC. Until now little information was available concerning either their expression or recognition by the immune system. Here we report that these non-K/D genes can provoke antibody responses and be recognized by cytolytic T cells. Immunization of C3H mice with L cells transfected with class I genomic clones resulted in antisera that reacted preferentially with cells from strain B10.P (the gene donor). Thus, these genes can be expressed by L cells. These products were recognized by cytolytic T cells produced by mixed lymphocyte culture with B10.P stimulators. One gene, represented in clone lambda 3a, was chosen for further analysis. A restriction fragment length polymorphism, detected between B10.P (KpDp) and B10.F(14R) (KbDp) and between B10 (KbDb) and B10.F(13R) (KpDb), has enabled us to map the lambda 3a sequence to the D or Tla region. Restriction endonuclease mapping of the lambda 3a clone shows that the gene is intact and that, although many restriction sites are conserved, the gene in lambda 3a differs from other class I genes. When the lambda 3a clone was transfected into mouse L cells, a new product was expressed. Cells expressing this product (designated L3a cells) were killed by primary D-end reactive, allospecific cytolytic T lymphocytes. The L3a cells were unreactive with monoclonal antibodies specific for the Kp,Dp,Qa-2, Tla.3, and Tla.5 molecules. PMID- 2991931 TI - Isolation of infectious human T-cell leukemia/lymphotropic virus type III (HTLV III) from patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) and from healthy carriers: a study of risk groups and tissue sources. AB - Acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) are thought to be caused by human T-cell leukemia/lymphotropic virus type III (HTLV III). Since the fall of 1982, independent isolates of HTLV-III have been obtained in this laboratory, in collaboration with several clinical groups, from 101 AIDS and ARC patients and healthy donors at risk for AIDS. Most isolates were from peripheral blood T lymphocytes established in cell culture, but some were obtained from bone marrow, lymph node, brain tissue, and cell-free plasma and from cells associated with saliva, cerebrospinal fluid, and semen. Virus was isolated from approximately 50% of AIDS patients, 85% of ARC patients, and 30% of healthy individuals at risk for AIDS. The risk groups included homosexuals, promiscuous heterosexuals, i.v. drug users, recipients of blood or blood products, and spouses and offspring of AIDS patients and others at risk for AIDS. A high correlation was seen between persistent levels of serum antibody and the ability to isolate virus from patient or donor leukocytes. Immunologic and nucleic acid analysis demonstrated that the virus isolates were highly related, although substantial diversity was observed in the restriction enzyme cleavage patterns of those studied in detail. Biological analysis of cells from infected patients and donors as well as from normal peripheral blood mononuclear cells exposed to virus in vitro demonstrated that OKT4/Leu3a+ (helper/inducer) lymphocytes were preferentially infected and were subjected to a characteristic cytopathic effect. The availability of multiple isolates of virus from a number of different patients and donors will greatly facilitate the characterization of HTLV-III and the study of possible biological and/or biochemical variants of the virus responsible for the development of AIDS, ARC, and related diseases. PMID- 2991933 TI - Papillomavirus genomes in human cervical tumors: analysis of their transcriptional activity. AB - Four of six human cervical carcinoma biopsies were shown to contain the DNA of the human papillomavirus type 16 (HPV-16) covalently linked with the tumor cell DNA. HPV-16-specific mRNA species were observed in only one of the four tumors. No such sequences were found in the three other specimens in conditions that permitted the detection of less than one mRNA molecule per cell. It is concluded that maintenance of the malignant nature of these cervical tumors does not depend on the continuous transcriptional activity of HPV-16. PMID- 2991932 TI - High prevalence of antibodies to acquired immune deficiency syndrome (AIDS) associated retrovirus (ARV) in AIDS and related conditions but not in other disease states. AB - A rapid, sensitive indirect immunofluorescence assay has been developed for detection of antibodies to the acquired immune deficiency syndrome (AIDS) associated retrovirus (ARV). The human T-cell line HUT-78 was chronically infected with ARV-2 and used to detect antibodies to virus-specific cytoplasmic antigens. Because the helper T-cell marker Leu-3 is substantially reduced in this cell line after ARV infection, it appears to be an important receptor for virus infection. Nearly all patients with AIDS and most cases with related conditions showed antibodies against ARV. Some healthy individuals in risk groups for developing AIDS also had antibodies to the agent. In contrast, no antibodies to the virus were found in any individuals outside the risk groups for developing AIDS or with diseases other than those associated with AIDS. The titers of antibodies to ARV and to Epstein-Barr virus varied independently from each other. The level of anti-ARV antibodies in a patient's serum was found to reflect the severity of the disease; it was lower in individuals with more severe manifestations. Taken together, these data support the role of ARV in AIDS and its related disorders. PMID- 2991934 TI - Demonstration of benzodiazepine-like molecules in the mammalian brain with a monoclonal antibody to benzodiazepines. AB - An anti-benzodiazepine monoclonal antibody has been used to demonstrate the existence of benzodiazepine-like molecules in the brain. Immunocytochemical experiments show that these molecules are neuronal and not glial and that they are ubiquitously distributed throughout the brain. Immunoblots indicate the presence of benzodiazepine-like epitopes in several brain peptides. Small benzodiazepine-like molecules were isolated from the brain soluble fraction by immunoaffinity chromatography. They block the binding of agonists, inverse agonists, and antagonists to the neuronal-type benzodiazepine receptor. The neurotransmitter gamma-aminobutyric acid increases the affinity of the benzodiazepine receptor for the purified endogenous molecules. The results indicate that the immunoaffinity-purified molecules behave like the neuronal-type benzodiazepine receptor agonists. The purified molecules, however, do not inhibit the binding of tritiated Ro 5-4864 to the "peripheral-type" benzodiazepine receptor. The results demonstrate the existence of benzodiazepine-like molecules in the brain that bind to the benzodiazepine receptor. These molecules are different from the endogenous benzodiazepine receptor ligands reported by others. PMID- 2991935 TI - The Abelson murine leukemia virus oncogene. PMID- 2991936 TI - Nucleosides and lymphocytes--an overview. PMID- 2991937 TI - Inosine 5'-monophosphate vs inosine and hypoxanthine as substrates for purine salvage in human lymphoid cells. AB - The ability of inosine 5'-monophosphate vs inosine or hypoxanthine to supply the total purine requirements of mitogen-stimulated human T cells or rapidly dividing human B lymphoblastoid cells was evaluated. Mitogen-stimulated human peripheral blood T cells were treated with aminopterin to inhibit purine synthesis de novo and make the cells dependent upon an exogenous purine source. Thymidine was added as a source of pyrimidines. Under these conditions, 25 microM inosine 5' monophosphate, inosine, and hypoxanthine showed comparable abilities to support [3H]thymidine incorporation into DNA at rates equal to that of untreated control cultures. In parallel experiments with the rapidly dividing human B lymphoblastoid cell line, WI-L2, treatment with aminopterin (plus thymidine) inhibited the growth rate by greater than 95%. The normal growth rate was restored by the addition of 30 microM inosine 5'-monophosphate, inosine, or hypoxanthine to the medium. However, in similar experiments with cell line No. 1254, a derivative of WI-L2 which lacks detectable ecto-5'-nucleotidase activity, only inosine and hypoxanthine (plus thymidine), but not inosine 5'-monophosphate (and thymidine) were able to restore the growth inhibition due to aminopterin. These results show that the catalytic activity of ecto-5'-nucleotidase is sufficient to meet the total purine requirements of mitogen-stimulated human T cells or rapidly dividing human B lymphoblastoid cells and suggest that this enzyme may have functional significance when rates of purine synthesis de novo are limited and/or an extracellular source of purine nucleotides is available. PMID- 2991938 TI - Purine metabolizing enzymes of lymphocyte cell populations: correlation between AMP-deaminase activity and dATP accumulation in murine lymphocytes. PMID- 2991939 TI - Metabolism and selectivity of arabinonucleoside in human lymphoid cells. AB - The selective toxicity of purine deoxynucleosides against lymphoid cells appears to be mediated by a preferential accumulation of the corresponding triphosphates in these cells. We report a study of the metabolism and toxicity of arabinonucleosides of guanine and cytosine toward human T- and B-lymphoblastoid cell lines. Both compounds inhibited the growth of T lymphoblasts at concentrations less than 2 microM. However, only ara-G exhibited a strong selectivity for T lymphocytes as indicated by a 100-fold greater toxicity to T than B cells. ara-G is not significantly degraded to guanine but is metabolized to the triphosphate. In common with the other arabinonucleoside, cytotoxicity by ara-G was associated with specific inhibition of DNA synthesis in cells. The capacity of T cells (CCRF-CEM) to accumulate ara-GTP was dependent primarily on deoxycytidine kinase. The level of intracellular ara-GTP accumulated after incubation with the corresponding nucleoside was 20- to 40-fold higher in T cells than either of two B-lymphoblast-cell lines, WI-L2 or PF-2S. The levels of phosphorylating activity for ara-C in extracts of T- and B-cell lines were approximately equal; in contrast, ara-G phosphorylating activity was four- to fivefold higher in B lymphoblasts. After removal of arabinonucleosides from the culture medium, ara-GTP levels in B lymphoblasts declined at a rate that was two to four times faster than that of ara-CTP. In marked contrast, no catabolism of the arabinonucleoside triphosphates was detected in T lymphoblasts. These results suggest that the selectivity of arabinonucleosides to human lymphoid cells of various phenotypes can be correlated with their nucleotide metabolism. The selectivity of ara-G for T and B cells can be correlated with their differential ability to catabolize ara-GTP. PMID- 2991940 TI - Regulation of cyclic AMP accumulation in lymphoid cells. AB - We have examined several features of the regulation of cyclic AMP accumulation in lymphoid cells isolated from peripheral blood of human subjects and in the murine T-lymphoma cell line, S49, S49 cells are unique because of the availability of variant clones with lesions in the pathway of cyclic AMP generation and response. We found that human lymphoid cells prepared at 4 degrees C showed substantially greater cyclic AMP accumulation in response to histamine and the beta-adrenergic agonist isoproterenol than did cells prepared at ambient temperature. The muscarinic cholinergic agonist carbamylcholine and peptide hormone somatostatin failed to inhibit cyclic AMP accumulation in human lymphoid cells and treatment with pertussis toxin (which blocks function of Gi, the guanine nucleotide binding protein that mediates inhibition of adenylate cyclase) only minimally increased cyclic AMP levels in these cells. Thus the Gi component of adenylate cyclase appears to play only a small role in modulating cyclic AMP levels in this mixed population of lymphoid cells. Incubation of whole blood with isoproterenol desensitized human lymphocytes to subsequent stimulation with beta agonist. This desensitization was associated with a redistribution of beta-adrenergic receptors such that a substantial portion of the receptors in intact cells could no longer bind a hydrophilic antagonist. Wild-type S49 lymphoma cells showed a similar redistribution of beta-adrenergic receptors after a few minutes' incubation with agonist. Based on studies in S49 variants, this redistribution is independent of components distal to receptors in the adenylate cyclase/cyclic AMP pathway. By contrast, a more slowly developing, agonist-mediated down-regulation of beta adrenergic receptors was blunted in variants with defective interaction between receptors and Gs, the guanine nucleotide binding protein that mediates stimulation of adenylate cyclase. Unlike results in human lymphoid cells, S49 cells show a prominent inhibition of cyclic AMP accumulation mediated by Gi; this inhibition is promoted by somatostatin and blocked by pertussis toxin. Inhibition by Gi is unable to account for the marked decrease in ability of the diterpene forskolin to maximally stimulate adenylate cyclase in S49 variants having defective Gs. These results emphasize that both Gs and Gi component are important in modulating cyclic AMP accumulation and receptors linked to adenylate cyclase in S49 lymphoma cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2991941 TI - Biological activity of phenolic compounds. Hepatic cytochrome P-450, cytochrome b5, and NADPH cytochrome c reductase in chicks and rats fed phenolic monomers, polymers, and glycosides. AB - Eight experiments were conducted to determine effects of a phenolic polymer (Kraft wood lignin, Indulin), phenolic glycosides (cane molasses and wood molasses), and phenolic monomers (vanillin, vanillic acid, ferulic acid, and p coumaric acid) on liver cytochromes P-450, cytochrome b5, and NADPH cytochrome c reductase in chicks and rats. Chicks fed 6.0% lignin had a higher (P less than 0.01) cytochromes P-450 content than did chicks fed 0% fiber, 6.0% wood cellulose (Solka Floc), or 6.0% arenaceous flour. NADPH cytochrome c reductase activity was not affected by treatment. Chicks fed 12.0% wood molasses had a higher (P less than 0.06) cytochromes P-450 level than did chicks fed 0% fiber or 6.0% wood molasses. Cane molasses incorporated at both 6.0 and 12.0% of the diet induced (P less than 0.05) cytochromes P-450 content over those of control-fed birds. Chicks fed 6.0% lignin, with or without antibiotic (bacitracin:neomycin sulfate, 2:1), had a higher (P less than 0.01) cytochromes P-450 level than did chicks fed control diets, with or without antibiotic. Additionally, chicks fed 6.0% lignin had lower (P less than 0.01) intestinal diaminopimelic acid (DAP) levels than did chicks fed 0% fiber. Rats fed 0% fiber, 6.0% wood cellulose, 6.0% arenaceous flour, or 6.0% lignin exhibited no difference in cytochrome level or activity among treatments. Chicks fed 0.5% vanillin, 0.5% vanillic acid, 0.5% ferulic acid, or 0.5% p-coumaric acid had comparable cytochromes level and activity compared with chicks fed no phenolics. Chicks fed 0.5% p-coumaric acid had lower (P less than 0.05) rates of gain than did chicks fed control or other phenolic containing diets. Rats fed these phenolics had similar cytochromes P-450 content among treatments. PMID- 2991942 TI - Spread of multiresistant gram-negative bacilli among severely immuno-compromised mice during prophylactic treatment with different oral antimicrobial drugs. AB - Gram-negative infections are generally regarded as a major cause of severe infections in granulocytopenic patients. To prevent such infections, selective decontamination (SD) of the digestive tract has been introduced. It is assumed that if SD is applied on all patients in an oncologic station it may not only provide protection to patients thus treated, but it may have also epidemiologic consequences. SD may considerably reduce the number of circulating (antibiotic resistant) strains in the ward and therewith significantly further reduce the chance of severe infection during periods of granulocytopenia. Because the effect which different kinds of antibiotics may have on the accumulation and spread of resistant bacteria cannot be studied in patients, it was investigated experimentally. Groups of 15 conventional mice were lethally irradiated and each group was treated with one antibiotic. On the day of irradiation one mouse per cage was orally contaminated with a multi-resistant strain either of E. coli or Ps. aeruginosa. The spread of these strains among cage mates as well as their effect on mortality was investigated. It appeared that ampicillin promoted spread and enhanced mortality in comparison with a control group, whereas antibiotics currently used for SD did prevent spread of the multi-resistant strain and reduced mortality. PMID- 2991943 TI - Selective decontamination of the digestive tract for the prevention of infection in acute leukemia. PMID- 2991944 TI - Prolonged granulocytopenia due to selective antimicrobial modulation of the intestinal flora: bias or biology? PMID- 2991945 TI - Effect of phenoxybenzamine on indomethacin induced systemic vasoconstriction in dogs. AB - The effect of phenoxybenzamine (PBZ) on indomethacin (IM) induced systemic vasoconstriction was studied in 22 dogs using a right heart bypass procedure which allowed for control of cardiac output. Indomethacin alone (n = 8) caused an increase in mean arterial pressure (MAP) from 71 +/- 6.5 mmHg to 116 +/- 7.6 mmHg (p less than .05). In PBZ pretreated dogs (n = 8), the increase in MAP was from 49 +/- 3.4 mmHg to 80 +/- 6.8 mmHg (p less than .05). In 3 dogs given PBZ alone, MAP fell from 81 +/- 7.3 mmHg to 42 +/- 3.8 mmHg (p less than .05). In 3 untreated dogs, MAP fell from 86 +/- 4.1 mmHg to 63 +/- 9.0 mmHg. Thus, IM causes vasoconstriction in the systemic circulation of the dog, and this effects does not appear to be mediated by alpha adrenergic stimulation. PMID- 2991946 TI - Studies on the mechanism of central cardiovascular and temperature responses to prostaglandin D2. AB - Administration of prostaglandin D2 (PGD2) into the left lateral cerebral ventricle (i.c.v.) of urethane anaesthetized rats caused increases in blood pressure, heart rate and body temperature. Pretreatment of the animals with the alpha-receptor blocking drug prazosin completely prevented the PGD2-induced rise in blood pressure, but did not affect the chronotropic and hyperpyrexic effects of PGD2. Pretreatment of the rats with the beta-receptor blocking drug propranolol completely suppressed the increase in heart rate, augmented the rise in blood pressure, but reduced the temperature increase by more than 50%. These data indicate that central activation of the sympathetic nervous system mediates the cardiovascular and important parts of the temperature effects of i.c.v. injected PGD2. The peripheral vasoconstriction (due to the stimulation of sympathetic alpha-receptors) causing the blood pressure increase is obviously of minor importance for the rise in body temperature. In contrast the enhanced production of heat by several organs following beta-receptor activation seems to be an important factor for the temperature increase. PMID- 2991947 TI - Polymorphonuclear leukocyte 5-lipoxygenase activity in psoriasis. AB - 5-lipoxygenase-derived products of arachidonic acid are implicated in the pathophysiology of psoriasis, a common hyperproliferative and inflammatory skin disease. We therefore examined whether there is an activation of this enzymatic pathway in extracutaneous tissues. For this purpose, we measured the conversion of 14C-arachidonic acid by polymorphonuclear leukocytes from psoriatic patients and controls. No significant difference in the generation of leukotriene B4 and 5 hydroxyeicosatetraenoic acid by polymorphonuclear leukocytes was noted between the two groups. We conclude that in psoriasis there is no enhanced activity of the 5-lipoxygenase pathway in circulating polymorphonuclear leukocytes. PMID- 2991948 TI - Effects of triethyl lead on hot-plate responsiveness and biochemical properties of hippocampus. AB - Rats treated with a single dose of triethyl lead chloride (TEL) by subcutaneous injection (7.9 mg/kg) showed a transient increase in latencies to lick the hind paw during hot-plate testing. The time course of triethyl lead-induced antinociception was temporally associated with depressed binding capacity of benzodiazepine receptor sites and reduced levels of Substance P. Both of these changes appeared to be confined to the hippocampus and were not apparent in the cortex or striatum of treated rats. Met-enkephalin levels were not altered in any region studied at any time during the 21-day postdosing period. Lead levels within the brain were higher than blood levels 1 week after triethyl lead injection. Although changes in more than one factor may account for the antinociceptive effect of triethyl lead, the hippocampus seems especially vulnerable to this amphiphilic organometal. PMID- 2991949 TI - Opiate antagonism reduces placentophagia and pup cleaning by parturient rats. AB - Since endogenous opiate mechanisms are activated during parturition, the present study examined in rats the effects of opiate antagonism on maternal care during and shortly after parturition. Endogenous opiate mechanisms were blocked in late pregnant rats by (1) naltrexone pellet implants (Experiment 1); (2) acute naloxone injections of 10 mg/kg (Experiment 2) or 0.1 mg/kg (Experiment 7); or (3) induction of opiate tolerance (Experiment 3). All methods resulted in a significant decrease in placentophagia and/or in cleaning pups of umbilical cords and birth fluids (Experiment 6). Other aspects of maternal care appeared relatively unaffected and 24 hr pup survival rats were lowered only by induction of morphine tolerance (probably via its effects on the young). In nonpregnant females, naloxone produced a small but significant decrease in placentophagia (Experiment 4) whereas morphine-tolerant nonpregnant females consumed placentas as readily as controls (Experiment 5). Thus the inhibition of placentophagia produced by opiate antagonism may be specific to conditions associated with parturition. These findings suggest that endogenous opiates support placenta eating and pup cleaning during and immediately after birth. Mediation may be via opiate effects on ingestive behavior, and/or via a reduction in the stress of parturition which otherwise can interfere with the female's ability to perform these tasks. PMID- 2991950 TI - Involvement of dopamine in the aversive stimulus properties of cocaine in rats. AB - Previous studies of cocaine self-administration have demonstrated central dopaminergic involvement in cocaine's positive reinforcing properties. The present study reports the ability of pimozide, a dopamine receptor antagonist, to attenuate a conditioned taste aversion induced by repeated injections of cocaine. Rats placed on a daily water deprivation schedule were subsequently presented with a novel saccharin taste in their drinking fluid immediately followed by administration of four 9 mg/kg injections of cocaine spaced at 20 min intervals. These animals exhibited a reduction in saccharin intake on subsequent presentations. Animals pretreated with pimozide 90 min prior to the saccharin cocaine pairings failed to show this reduction. In a second experiment using an identical procedure, repeated injections of lithium chloride were shown to induce a CTA both in pimozide-pretreated and control animals. The results of these two experiments are consistent with the notion that a functional relationship may exist between neurochemical mechanisms underlying both the aversive (CTA inducing) and positive reinforcing properties of self-administered drugs such as cocaine. PMID- 2991951 TI - Changes in brain opiate receptors in rats with isolation syndrome. AB - Prolonged social isolation of Wistar rats produces various neurochemical, neuroendocrine and behavioral changes. Some of isolated rats exhibit interspecies aggressive behavior (mouse-killing). It is suggested that social isolation leads to a stress-like state of rats. On the other hand, the participation of endogenous opioid mechanisms in stress is well known. We found that social isolation induced a decrease in the number of opiate receptors in rat frontal cortex, striatum, hippocampus and periaqueductal grey matter. The decrease was most pronounced in isolated aggressive rats. The affinity of opiate receptors in striatum and hippocampus was increased and in frontal cortex was decreased, these changes being more distinct in isolated aggressive than in isolated non aggressive rats. These data suggest that social isolation of rats induces great changes in brain opiate receptor functions. PMID- 2991952 TI - Significance of serotonin in central nervous functions. PMID- 2991954 TI - Receptor-mediated regulation of 5-hydroxytryptamine metabolism: current knowledge and open questions. AB - As demonstrated for the catecholamine system several receptor-mediated mechanisms appear to be involved in the regulation of serotonergic transmission, though regulatory processes of serotonergic transmission are less well investigated as compared with the catecholamine system. One of the main homeostatic mechanisms appears to be neuronal feedback mediated by postsynaptic 5-HT receptors. The constituents of the neuronal feedback loop are unknown at present. In addition to neuronal feedback, an autoreceptor-mediated feedback appears to exist. According to their localization, 5-HT autoreceptors can be subdivided into somatodendritic and presynaptic autoreceptors. Stimulation of somatodendritic 5-HT receptors leads to inhibition of the firing rate, and stimulation of presynaptic autoreceptors in vitro to inhibition of impulse-induced release of 5-HT. Further to 5-HT receptors, other transmitter receptors appear to exist on presynaptic nerve terminals which may either inhibit or facilitate 5-HT release. The physiological significance of all these presynaptic receptors remains to be shown. The evaluation of feedback mechanisms controlling 5-HT neurons is hampered by the fact that no selective central 5-HT antagonists are available at present. There is an urgent need for selective receptor antagonists for 5-HT autoreceptors as well as postsynaptic receptors. Binding studies have revealed at least two subtypes of 5-HT receptors, 5-HT1 and 5-HT2 receptors. Selective 5-HT1 and 5-HT2 agonists and antagonists are needed in order to characterize these receptors subtypes and to find their functional correlates. PMID- 2991953 TI - Anatomical features and physiological properties of central serotonin neurons. AB - The authors describe the anatomical features and physiological properties of central serotonergic neurons. The central serotonin neurons (part of which store peptides [substance P, TRF, enkephalins] in addition to 5HT) are highly collateralized reticular-type brain stem neurons receiving multi-modal afferent information from ascending sensory and descending motor pathways. They are under control by noradrenergic, peptidergic and and gaba-ergic projection neurons and interneurons. Furthermore, they establish variable synaptoid and synaptic contacts to neuronal, glial and secretory targets throughout the entire neuraxis and send terminal branches into the ventricular CSF space. Firing rate and transmission activity appear to be controlled in a complex and rather rigid manner by 5HT release-dependent dendrodendritic and dendrosomatic inhibition via autoreceptors (which also regulate release at the axon terminals) and via transsynaptic inhibitory feedback circuits which may involve gabaergic projection and interneurons. 3H-imipramine appears to bind to an "imipramine recognition site" in the vicinity of the 5HT carrier, and to a variety of other transport and (postsynaptic) receptor sites (NA uptake, H1, 5HT2- and alpha 1-binding sites). Circumstantial evidence points to an as yet undetermined role of the postsynaptic 5HT-1-binding sites in neurotransmission. 5HT-2-binding sites fulfil the criteria for receptors: binding affinity of antagonists to these sites correlates significantly with their potency to inhibit behavioral excitation in rats elicited by 5-hydroxytryptophan or 5HT agonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2991955 TI - Serotonin involvement in lisuride-induced mounting and in sleep. AB - Mounting behavior of rats induced by several drugs, such as e.g. p-CPA, 5,7 dihydroxytryptamine, L-DOPA or lisuride, appears to result from a combined decrease of 5-hydroxytryptamine (5-HT) and increase of dopamine (DA) neurotransmission. In this paper, lisuride-induced mounting is proposed as a behavioral model for detecting pharmacologically active drugs that interact with monoaminergic mechanisms, e.g. 5-HT reuptake blockers and type A monoamine oxidase inhibitors. With regard to sleep, 5-HT appears to sustain a relevant part in controlling the sleep-wakefulness cycle. However, other transmitters or neuromodulators (catecholamines, oligopeptides etc.) may also be involved in sleep mechanisms. PMID- 2991956 TI - Photogeneration of superoxide and decarboxylated peptide radicals by carboquone, mitomycin C and streptonigrin. An electron spin resonance and spin trapping study. PMID- 2991957 TI - A 6-year experience with immediate reconstruction after mastectomy for cancer. AB - A 6-year retrospective review is presented of 185 patients who underwent immediate reconstruction of the breast at the same operation as mastectomy for carcinoma. The patients were treated at two institutions under similar protocols of patient selection, surgical technique, and postoperative care. A detailed evaluation is presented from both the oncologic and surgical points of view. The data support the conclusion that immediate reconstruction of the breast does not alter survival or cancer recurrence rates and does not interfere with the treatment of primary or secondary disease. A low incidence of significant surgical complications is also detailed. Combined with previous reports answering psychological concerns about this mode and timing of reconstruction, this review offers significant reassurance about the overall safety of immediate reconstruction. The authors therefore recommend immediate reconstruction of the breast as a safe treatment option for the woman facing mastectomy. PMID- 2991958 TI - [Electrophysiologic studies of alcohol dependent patients]. AB - Electrophysiological methods were used to examine 52 ward patients with a long history of alcohol abuse. Alertness fluctuations were much more common in the EEG than among the healthy control subjects. 86.5% of the ENG findings were abnormal and indicated demyelinisation in most cases. The latence times of the brain stem potentials were longer than normal in about half of the patients and were definitely pathological in 13.5%. There was no statistical correlation between drinking habits and the results of the examination. Unknown pathogenetic factors are therefore suspected. The polytopic pattern of damage necessitates the use of a broad range of signostic techniques. PMID- 2991959 TI - Effects of the contraceptive pill on sedative responses to clonidine and apomorphine in normal women. AB - In normal women sedation following intravenous administration of the alpha 2 adrenoceptor agonist clonidine (1.3 micrograms/kg) was significantly greater following 3 weeks' continuous treatment with the combined contraceptive pill than at the end of the 7-day withdrawal period. In contrast, sedative responses to the dopamine agonist apomorphine (5 micrograms/kg subcutaneously) were decreased when subjects were taking the contraceptive pill. These findings are in agreement with animal experimental studies, indicating that oestrogen can alter alpha adrenoceptor and dopamine autoreceptor sensitivity. Such interactions may be involved in the effects of female sex steroids on mood. PMID- 2991960 TI - Purine nucleoside--mediated immobility in mice: reversal by antidepressants. AB - In the forced swimming-induced immobility (despair) test model, adenosine, and 2 chloroadenosine treatment prolonged the immobilization period in mice. Dipyridamole, which is known to inhibit adenosine uptake, potentiated the adenosine effect. The purinoceptor antagonists caffeine and theophylline blocked purine nucleoside-induced enhancement of immobilization. Tricyclic antidepressants such as imipramine and desipramine, the MAO inhibitor tranylcypromine, and amphetamine, a psychostimulant, reversed purine nucleoside induced immobility. On the other hand, quipazine, fluoxetine, and amitriptyline failed to reverse purine nucleosides-induced prolongation of immobility. None of the antidepressants in the doses investigated had any effect by themselves. Reserpine also prolonged forced swimming-induced immobility in mice. The antidepressants fluoxetine and quipazine, but not methylxanthine pretreatment, reversed reserpine-induced immobility in this test model. These results indicate that adenosine and 2-chloroadenosine probably reduce norepinephrine outflow through their action on presynaptic purinoceptors on noradrenergic neurons and thereby cause prolongation of immobility in animals. PMID- 2991962 TI - A beta-carboline antagonizes benzodiazepine actions but does not precipitate the abstinence syndrome in cats. AB - The beta-carbolines, which are potent ligands for benzodiazepine receptors, antagonize the pharmacological actions of benzodiazepines. In the cat, the stable beta-carboline derivative methylamide-beta-carboline-3-carboxylate, FG 7142, and the specific benzodiazepine antagonist Ro 15-1788 reversed behavioral and electroencephalographic (EEG) changes produced by a single dose of diazepam. Surprisingly, the beta-carboline did not elicit signs of withdrawal when given after 22 days of a daily dose regimen of diazepam, while Ro 15-1788 precipitated an acute abstinence syndrome largely characterized by tremors, increased muscle tone, back arching, myoclonic jerks and pupil dilatation. Unlike Ro 15-1788, the beta-carboline produced effects of its own such as behavioral states of arousal and fearfulness. These findings indicate that the beta-carboline functionally interacts with benzodiazepine receptors in a manner unlike that seen with typical agonists and antagonists. PMID- 2991961 TI - Locomotor effects of lisuride: a consequence of dopaminergic and serotonergic actions. AB - The open-field test was used to study the involvement of serotonergic and dopaminergic mechanisms in the action of lisuride on locomotor activity in the rat. Lisuride produced a biphasic locomotor effect. The maximum locomotor stimulatory response of lisuride was stronger than that of apomorphine and comparable with that of apomorphine and LSD combined. Hypermotility induced by high doses of lisuride was partially suppressed by the serotonin antagonist cyproheptadine and not further enhanced by LSD. A moderate dose of lisuride potentiated apomorphine-induced hypermotility in the same manner as has been shown for LSD. Lesion of dopaminergic structures within the median raphe nucleus by 6-OHDA produced a potentiation of lisuride-induced hypermotility. This effect was suppressed by cyproheptadine. The locomotor inhibitory effect of low doses of lisuride may be related to a stimulation of presynaptic mesolimbic dopamine receptors. It is concluded that the locomotor stimulant effect of higher doses of lisuride may depend on stimulation of postsynaptic dopamine receptors and a serotonergic action and that the locomotor effects of lisuride reflect a complex interaction at dopaminergic and serotonergic transmission systems. PMID- 2991963 TI - Responses of prolactin and growth hormone to L-tryptophan infusion: effects in normal subjects and schizophrenic patients receiving neuroleptics. AB - Intravenous infusion of L-tryptophan (LTP) in 18 normal subjects produced a significant increase in plasma prolactin (PRL), growth hormone (GH), and self ratings of drowsiness. There was no correlation between the PRL and GH responses, or between the hormonal responses and drowsiness. Saline infusion did not result in endocrine or psychological changes. The effect of LTP on both PRL and GH was dose-related in that LTP 7.5 g produced greater endocrine responses than 5.0 g. It was not significantly decreased by cyproheptadine, a 5-HT receptor antagonist. Schizophrenic patients receiving neuroleptics had increased PRL response to LTP, possibly because of the drug-induced disinhibition of PRL release. Their GH response to LTP was markedly decreased. The mechanism of this effect requires further investigation. PMID- 2991965 TI - Diazepam administration to mice prevents some of the changes in monoamine mediated behaviour produced by repeated electroconvulsive shock treatment. AB - Administration to mice of electroconvulsive shock (ECS) five times over 10 days results in an enhanced 5-HTP-induced head twitch response, an enhanced apomorphine-induced locomotor response and an attenuated sedation response to the alpha 2-adrenoceptor agonist clonidine. Diazepam (1.25 mg/kg IP) injected 5 min before each ECS abolished the enhanced 5-HT- and dopamine-mediated responses but left the attenuated sedation response unaltered. When diazepam was given 5 min after each convulsion it still had the same effect, although its effects on the ECS-induced changes was blocked by administration of the specific benzodiazepine antagonist Ro 15-1788 (10 mg/kg IP) at the same time as diazepam. It is concluded that diazepam can abolish the ECS-induced changes in 5-HT- and DA-mediated behaviour, but not alpha 2-adrenoceptor-mediated responses, possibly by interfering with post-ictal changes. The implications for administration of benzodiazepines during electroconvulsive therapy (ECT) are discussed. PMID- 2991964 TI - Yawning elicited by systemic and intrastriatal injection of piribedil and apomorphine in the rat. AB - The behavioural effects of systemic and intrastriatal injections of the dopamine agonists piribedil and apomorphine in male rats were examined. Bilateral application of piribedil (50 and 100 micrograms) or apomorphine (5, 10 and 20 micrograms) to the striatum produced yawning and chewing mouth movements accompanied by intermittent stretching and sexual arousal. Low doses of piribedil (1.25 and 2.5 mg/kg) and apomorphine (0.1 and 0.2 mg/kg) injected SC produced an identical yawning syndrome. Previous work has suggested that yawning elicited by systemic dopamine agonist treatment is a consequence of dopamine autoreceptor stimulation. Similarly, the most likely explanation of the present data is that yawning elicited by systemic and central dopamine agonist treatment was due to activation of dopamine autoreceptors. Systemic injection of haloperidol and scopolamine abolished yawning induced by intrastriatal piribedil and these data provide tentative support for the proposal that a dopamine-acetylcholine link may be involved in the expression of yawning. PMID- 2991966 TI - Dopaminomimetic action of diphenylhydantoin in rat striatum: effect on homovanillic acid and cyclic AMP levels. AB - Diphenylhydantoin (DPH) is known to induce reversible paroxysmal dyskinesias and paranoid psychosis in humans, but its interactions with dopamine (DA) metabolism are not clear. Single doses of DPH (60-100 mg kg-1), with serum levels over 10 micrograms ml-1, reduced homovanillic acid (HVA) levels in rat striatum. The DPH induced HVA decrease was enhanced by supersensitivity of postsynaptic DA receptors following chronic haloperidol (Hal) administration. DPH 60 mg kg-1 given acutely enhanced the HVA decrease induced by apomorphine (Apo) and partially counteracted the HVA increase following acute Hal (0.1-0.5-2 mg kg-1). After chronic DPH treatment, Apo was ineffective in reducing striatal HVA levels. Concomitant chronic treatment with DPH and Hal counteracted the development of supersensitivity of postsynaptic DA receptors to Apo. Single doses of DPH (30-60 100 mg kg-1) increased cyclic AMP striatal content; this effect was blocked by Hal. A dopaminomimetic DPH action and a subsensitivity of postsynaptic DA receptors after chronic DPH seem to be suggested. These effects could be related to the dyskinetic and psychotic syndromes produced by the drug. PMID- 2991967 TI - Blockade of nucleus accumbens opiate receptors attenuates intravenous heroin reward in the rat. AB - A quaternary derivative of naloxone, methyl naloxonium chloride (MN), was administered intracerebrally to rats trained to self-administer heroin intravenously. Increases in intravenous (IV) heroin self-administration rates were found following injections of low doses of MN into the nucleus accumbens (N.Acc), but not following injections of low doses of MN into the ventral tegmental area (VTA). These results were interpreted to suggest that the rewarding properties of IV heroin were decreased following N.Acc opiate receptor blockade. The relative insensitivity of the VTA to MN treatment was taken to suggest that VTA opiate receptors are either not essential or play a secondary role in mediating IV heroin self-administration. The present data support the notion that post-synaptic N.Acc opiate receptors play a crucial role in maintaining IV heroin self-administration. PMID- 2991968 TI - Subsensitivity to muscimol-induced catalepsy after long-term administration of phenytoin in rats. AB - Male albino rats were injected with 25 mg/kg of phenytoin (PHT) every day for 20 consecutive days and were tested on days 21 and 28 for their response to 1 or 2 mg/kg of muscimol, a GABA receptor agonist. Rats treated with PHT showed a decreased responsiveness to muscimol-induced catalepsy (2 mg/kg) on day 21 but not on day 28. Acutely administered PHT, on the contrary, had a tendency to potentiate muscimol-induced catalepsy. Muscimol-induced catalepsy was not antagonized by acute treatment with bicuculline (0.5-2.0 mg/kg). It is proposed that withdrawal after long-term administration of PHT reduces the sensitivity of a GABA receptor site not sensitive to bicuculline. PMID- 2991969 TI - The Public Health Service role in the disposal of chemical munitions. AB - Within the last decade, the Centers for Disease Control (CDC) has increasingly emphasized environmental public health activities. The Center for Environmental Health (CEH), one of nine major units of the CDC, was established as a focus for assessment and prevention of environmentally related diseases. Many new, legislatively mandated programs have been delegated to CEH. One such mandated responsibility in Public Laws 91-121 and 91-441 directs the Department of Health and Human Services or its designee to review the Department of Defense (DOD) plans to dispose of or to transport chemical warfare agents. The Chemical Munitions Demilitarization Program, CEH, reviews DOD plans and makes recommendations to ensure that hazards to public health and safety have been provided for in the plans. In addition, these CEH staffers periodically review approved activities at DOD facilities, assessing their monitoring and evaluation programs. CEH staffers also contact State and local health and environmental agencies to identify and evaluate any concerns of the agencies or the public relating to these activities. PMID- 2991970 TI - Alcoholic skeletal myopathy, a clinical and pathological study. AB - One hundred and fifty-one inpatients with a history of chronic heavy alcohol intake were examined for evidence of muscle disease. Ninety-two patients (60 per cent) had histologically abnormal biopsies of the quadriceps muscle. The most common abnormality, which was often severe, was type II muscle fibre atrophy. Seven patients (5 per cent) had histological evidence of acute myopathy, one of whom presented with the full clinical picture of acute rhabdomyolysis. Twenty three patients had cirrhosis, 36 were significantly malnourished and 98 had evidence of a peripheral neuropathy. None of these features, however, were sufficient to account for the muscle abnormalities. There was no clear relationship between musculo-skeletal symptoms and muscle biopsy histology. Serum creatine kinase activity was elevated in only 23 subjects and was an insensitive indicator of subclinical acute myopathy and of chronic alcoholic myopathy. Follow up studies after abstinence from alcohol invariably showed both objective and subjective improvement of muscle function - often in the absence of any clinical recovery from the peripheral neuropathy. Continued alcohol consumption was accompanied by persistence and often deterioration of muscle fibre atrophy. It is concluded that chronic skeletal myopathy is a frequent consequence of alcohol abuse and may result from a direct toxic effect of ethanol on muscle fibres. PMID- 2991971 TI - Serum angiotensin converting enzyme in sarcoidosis: sensitivity and specificity in diagnosis: correlations with disease activity, duration, extra-thoracic involvement, radiographic type and therapy. AB - Serum angiotensin converting enzyme (SACE), despite certain limitations, has been found to be a useful adjunct to the diagnosis, assessment of disease activity, and management of sarcoidosis. The spectrofluorimetric assay was used to measure SACE in 50 normal controls, 76 patients with tuberculosis (42 pulmonary, 16 lymphatic and 18 military cases), 20 patients with cryptogenic fibrosing alveolitis, 50 patients with silicosis, three patients with extrinsic allergic alveolitis, 10 patients with Crohn's disease, two patients with Gaucher's disease, and 128 patients with sarcoidosis on 303 occasions (144 during periods judged as clinically active and 189 inactive). Our results show a normal range (mean +/- 2SD) of 19-54 nmol/ml/min. The false positive rate is 2 per cent in normal controls, 9.2 per cent in tuberculosis (38.9 per cent in military but 0 per cent in the other forms), 48 per cent in silicosis, 100 per cent in Gaucher's disease, and 0 per cent in the other diseases. The sensitivity, specificity, positive and negative predictive values were 58.1, 83.8, 83.8 and 58.1 per cent respectively. The sensitivity rose to 85.9 per cent if only those samples taken from patients in whom sarcoidosis was suspected on initial presentation were included, and 92.1 per cent if only those with clinically active sarcoidosis were included. The sensitivity of SACE as a diagnostic test in sarcoidosis is thus influenced by the relative frequency of active and inactive sarcoidosis. The specificity is influenced by the prevalence of military tuberculosis and silicosis, but is uninfluenced by other common varieties of interstitial lung disease such as cryptogenic fibrosing alveolitis and extrinsic allergic alveolitis, or by other non-pulmonary granulomatous disease. There was no correlation of the SACE level with age, sex, population group, associated other illness or duration of sarcoidosis. Statistically, SACE levels were significantly higher in patients with Types II and III chest radiographs as compared to Type I and 0, and also in those with additional clinically evident extra-thoracic disease e.g. in lymph node, eye and especially multiple systems. SACE also reflected clinical activity with levels being statistically significantly greater in those patients assessed as having active disease, although 7.9 per cent of these had normal levels. Our observations indicate that SACE levels correlate well with disease activity longitudinally, both in relation to spontaneous remission and steroid therapy, and are thus helpful in patient management. PMID- 2991972 TI - Late functional and biochemical changes in mouse lung after irradiation: differential effects of WR-2721. AB - The radioprotective effect of WR-2721 on late damage after whole thorax irradiation has been studied after split doses of radiation using the standard death and breathing rate assays at monthly intervals between 3 and 15 months after irradiation, as well as two biochemical measurements of injury at 15 months, hydroxyproline (HP), an indicator of tissue fibrosis, and DNA content, an indicator of tissue cellularity. A comparison of HP/lung and breaths per minute (BPM) in each dose group in the WR-2721 and non-WR-2721-treated mice 15 months after irradiation showed that the relationship between these two assays of late lung injury was not the same. There were large dose-related increases in breathing rate corresponding to relatively small changes in HP in the lungs of mice given radiation alone. In contrast, the mice given WR-2721 before irradiation showed large dose-related increases in HP/lung, but BPM remained relatively constant independent of dose. These data suggest then that changes in breathing rate and deaths later than 9 months after whole lung irradiation may not be due to collagen accumulation in the lung. WR-2721 did protect better against late lung functional changes (protection factors (PF) = 1.6) and late deaths (PF = 1.51) than against earlier changes in these same assays (PF = 1.4 and 1.28, respectively). Although the earlier-appearing injury after whole thoracic irradiation is most likely related to lung damage with deaths and increases in breathing rate resulting from pneumonitis, the cause of the late appearing functional injury in the lung after radiation is not clear. Thus protection of late lung damage measured from either lethality or breathing rate is not related to the prevention of lung fibrosis. PMID- 2991973 TI - The effect of dexamethasone on the cytotoxic and enzymatic response of cultured endothelial cells to radiation. AB - Experiments were conducted to determine (1) whether glucocorticoids directly protected endothelial cells (EC) from radiation and (2) if angiotensin converting enzyme (ACE) activity, known to be increased by glucocorticoid, played a role in the EC response to radiation. Confluent monolayers of EC cultured from bovine aorta EC were treated with dexamethasone (10(-6) M); after irradiation (5.0 Gy, 60Co gamma), ACE and lactate dehydrogenase (LDH) activities, DNA and protein contents, and nuclei number were measured. Twenty-four hours after 5 Gy, there was increased cell loss (-40%, P less than 0.001), greater LDH release (greater than 100%, P less than 0.001), more LDH activity per cell (+40%, P less than 0.001), and unchanged ACE activity compared to sham-irradiated control EC. However, 48 hr after 5 Gy, ACE activity per cell was decreased (-24%, P less than 0.005). A 48-hr exposure to dexamethasone alone was accompanied by a slight cell loss (-10%, P less than 0.001) and increased cellular ACE activity (+40-140%, P less than 0.001), but a 24-hr dexamethasone exposure was not cytotoxic and did not change ACE activity. Dexamethasone exposure for 48 hr before and after irradiation did not attenuate cell loss or LDH release. However, combined dexamethasone treatment and radiation increased cellular ACE activity at a time when neither agent alone had an effect (24-hr dexamethasone exposure before 5 Gy and assayed 24 hr after 5 Gy). This interaction between radiation and dexamethasone treatment suggests that the glucocorticoid modifies the cell's response to injury. Although this interaction does not ameliorate radiation cytotoxicity, maintenance of ACE levels in injured vessels by hormones may have physiological significance in the hemodynamics of irradiated tissues. PMID- 2991974 TI - [Cyclic nucleotide levels in the regenerating liver of rats following irradiation and protection by serotonin]. AB - Changes in cAMP and cGMP levels after X-irradiation and radioprotective treatment with serotonin were studied using a model of impulse inhibition and repair of transcription, translation and replication by cycloheximide in rat hepatocytes in vivo. Irradiation was shown to inhibit the cyclic nucleotide synthesis. Serotonin increased the contents of cAMP and cGMP maintaining their elevated level after X irradiation. PMID- 2991975 TI - [Cytogenetic effect of gamma quanta and secondary radiation generated by 70 GeV protons on Chinese hamster fibroblasts]. AB - The study of the cytogenetic effects (for instance, the chromosome aberration frequency in G2 cells and micronucleus formation) after exposure of Chinese hamster cells to gamma-rays and secondary radiation generated by 70 GeV protons showed that the relative biological effectiveness (RBE) of secondary radiation was approximately 3. The contribution of isochromatid deletions and exchanges into the total spectrum of rearrangements induced by the secondary radiation was different from that observed on gamma-irradiation. The absence of the modifying effect of caffeine on the cells exposed to the secondary radiation indicated that RBE of the secondary radiation from 70 GeV protons was mainly associated with its inhibitory action on the cytogenetic damages repair. PMID- 2991976 TI - [DNA damage and repair in Chinese hamster fibroblasts exposed to gamma and secondary radiation generated by 70 GeV protons]. AB - The cytochemical study of DNA damage and repair in a Chinese hamster fibroblast culture exposed to gamma-rays and secondary radiation from 70 GeV protons showed no significant differences between the two types of radiation. PMID- 2991977 TI - [Effect of ionizing radiation on the properties of rat liver phosphoprotein phosphatase]. AB - Phosphoproteidphosphatase (3.1.3.16) of high specificity for lysil-tRNA synthetase (6.1.1.6) and proteins of high-molecular-weight multienzyme complex of aminoacyl-tRNA-synthetases (6.1.1.) was isolated from rat liver. Irradiation of animals with an absolutely lethal dose of 0.21 C/kg decreased phosphoproteidphosphatase activity: a 3-4-fold decrease was noted 1 hr following irradiation. The activity of the enzyme isolated 24 hr after irradiation increased but did not reach the control level. PMID- 2991978 TI - [Action of a UHF field on GABA-ergic and acetylcholinergic systems in synaptic transmission]. AB - It was shown that exposure of rats to microwaves (800 MHz, 16 Hz modulation, 1-3 mW/cm2) decreased muscimol binding with synaptic membranes and reduced acetycholinesterase activity in the rat brain. PMID- 2991979 TI - [Computed tomographic diagnosis of facial bone tumors]. PMID- 2991980 TI - Tissue relaxation time: in vivo field dependence. AB - Relaxation times (T1 and T2) were measured in vivo in mongrel dogs at fields of 0.3, 0.5, 1.0, 1.35, and 1.5 tesla (T). T1 was measured using nine values of inversion time ranging from 10 to 1,280 msec. T2 was measured with a four-point multiple spin-echo sequence. Relaxation times were calculated for muscle, kidney cortex, spleen, and adipose tissue. T2 is independent of field. A linear fit to the field dependence of T1 yields slopes of 400-500 msec/T for tissues in which the primary source of protons is water. The lower slope of adipose (approximately 150 msec/T) reflects the different mechanism of spin-lattice relaxation of the CH2 protons. PMID- 2991981 TI - Enzymatic patterns of the germ cells in the adluminal compartment of the testis in mammals and man; a comparative study. PMID- 2991982 TI - Seasonal changes in the activity of phosphatases in Sertoli cells of the frog Rana temporaria. PMID- 2991983 TI - [Adaptation of distal tubular segments to changes in electrolyte metabolism]. PMID- 2991984 TI - The distributions of 5'-nucleotidase and adenosine A1 receptors: evidence for diversification and conservation in the hippocampi of several commonly employed experimental animals. PMID- 2991985 TI - [Metabolic zoning of the human liver parenchyma in comparison with various mammals]. PMID- 2991987 TI - Transforming growth factors. Isolation, characterization, and interaction with cellular receptors. PMID- 2991986 TI - Two distinct mechanisms involving growth factors employed in subversion of growth regulation by oncogenes. PMID- 2991989 TI - The possible role of a new lymphotropic retrovirus (LAV) in the pathogeny of AIDS and AIDS-related diseases. PMID- 2991988 TI - Genital papillomavirus infections. PMID- 2991990 TI - The human T-cell leukemia-lymphoma virus family. PMID- 2991991 TI - Approaching the control of primary liver cancer by means of a virus vaccine. PMID- 2991992 TI - Maternal transmission of resistance to virus-induced leukemia. PMID- 2991993 TI - [Mitochondrial biogenesis; import of nuclear-coded proteins into mitochondria]. PMID- 2991994 TI - The twelfth Bartlett memorial lecture: the role of single neurons in the psychology of perception. PMID- 2991995 TI - [Bronchiolo-alveolar carcinoma. Radiological and clinical diagnostic considerations]. AB - The authors present 42 cases of histologically confirmed bronchioloalveolar carcinoma. The purpose of the study is to investigate the possibility to improve bronchioloalveolar carcinoma diagnosis by radiological-clinical correlations. Results indicate the importance of clinical findings especially in the diffuse multinodular variety. PMID- 2991996 TI - [Nuclear spin tomography of the petrous bone and cerebellopontiLe angle. Methods and normal anatomy]. AB - Proton nuclear resonance tomography, also called magnetic resonance imaging, is a new imaging procedure which makes it possible to obtain tomographic cuts of the petrous bone in three planes. It is complementary to CT and is able to demonstrate the small structures of the inner ear and of the cerebello-pontine angle, without bone artefacts and without overlap or distortion. By using special surface coils combined with thin sections and a gradient-zoom technique, it is possible to obtain high resolution images which equal those of modern CT. By choosing suitable exposure parameters and a multi-echo technique, one can obtain excellent tissue contrast without using positive or negative contrast media. PMID- 2991998 TI - Computed tomography of pulmonary blastoma in a child. PMID- 2991997 TI - [Computed tomographic observation of a case of microlithiasis alveolaris pulmonum]. PMID- 2991999 TI - [Mediastinal lymphangioma--value of computed tomography]. PMID- 2992000 TI - [Mediastinal pancreatic pseudocyst]. PMID- 2992001 TI - [Thymus carcinoid]. PMID- 2992002 TI - [Computed tomographic diagnosis of a congenital optic nerve coloboma]. PMID- 2992003 TI - [Computed tomographic detection of a bilateral choroidal osteoma]. PMID- 2992004 TI - [Embolism-induced hemorrhagic cerebral infarct. Detection of the source of the embolism using B-scan sonography of the carotid bifurcation]. PMID- 2992005 TI - [Medullary carcinoma of the thyroid with calcified metastases]. PMID- 2992006 TI - [Spontaneous pneumoperitoneum without peritonitis]. PMID- 2992007 TI - [Clinical results of a simple new nuclear medical method for determining cerebral blood flow]. AB - Dynamic cerebral scintigraphy is a widely used, non-invasive procedure for the diagnosis of haemodynamic changes in the supra-aortic arterial territory. Eighty examinations have been carried out using a new method, which depends on demonstrating the time of arrival of the bolus within certain parameters. A definite advantage of the simultaneous demonstration of flow in various vascular territories is the fact that it is easily and rapidly perceived and that it is readily interpreted without depending on the subjective judgement of the operator. In addition, it is unnecessary to measure regions of interest. PMID- 2992008 TI - [Cerebral infarct following trauma]. AB - Vascular occlusions due to cerebral trauma have always been regarded as great rarities. However, we have found hypo-dense foci of vascular distribution in 3.5% of 3500 CT examinations for trauma during the late phase. Lesions in the vascular territory of the posterior cerebral artery are usually the result of supratentorial pressure rise from epidural and subdural haematomas, leading to compression of the vessels against the edge of the tentorium. Typical infarcts in the territory of the medial and anterior cerebral arteries were found only rarely by CT after cerebral trauma. Infarcts at the watersheds between the three vascular territories were found with surprising frequency and small infarcts were found in the basal ganglia. It is assumed that these were due to ischaemic or hypoxic events due to cardiac or pulmonary complications during the initial phase. PMID- 2992009 TI - [Technic for computed tomographic study of skeletal muscles in neuromuscular diseases]. AB - Computed tomography provides a complete view of morphological alterations in skeletal muscle caused by neuromuscular diseases. A good image quality of the lower as well as of the upper limbs is obtained by choosing a small scanning field and an appropriate scanning-position. The arms are best examined one by one. In follow-up studies muscular atrophy can be documented by planimetric and densimetric measurements. Furthermore CT-scanning is a convenient method for selecting suitable muscles for muscle biopsy. PMID- 2992010 TI - [Serial computed tomography. Value of densitometric measurements in the diagnosis of benign and malignant lesions of the liver, pancreas and kidney]. AB - By means of serial computer tomography it is possible to show changes in normal and abnormal tissues in the liver, pancreas, and kidney with characteristic contrast kinetics. Diagnosis can be improved by evaluating intra-vascular parenchymal contrast within the first minute after an intravenous bolus injection. A tissue diagnosis is not possible in the majority of lesions in the liver, but can usually be made in lesions of the pancreas and kidneys. PMID- 2992011 TI - [Visualization of the pancreatic ducts by ultrasound tomography in pancreatic diseases with bile duct obstruction]. AB - The sonographic findings in 274 patients with biliary obstruction and/or dilatation of the pancreatic duct of more than 4 mm were evaluated (128 common duct stones, 82 carcinoma of pancreas, 64 chronic pancreatitis). Carcinoma of the pancreatic head caused a greater degree of dilatation of the common bile duct (average diameter 18.3 mm) than did chronic pancreatitis (10.5 mm) or biliary stones (14.2 mm). Dilatation of the pancreatic duct greater than 4 mm occurred with approximately equal frequency with carcinomas (59%) and chronic pancreatitis (64%); it was found in only 1.6% of biliary duct stones. Dilatation of intra-and extra-hepatic bile ducts and hydrops of the gall bladder was significantly higher with carcinoma of the pancreas than in chronic pancreatitis. PMID- 2992012 TI - Comparison of ultrasonography and transhepatic cholangiography in the evaluation of obstructive jaundice. AB - The diagnostic value of ultrasonography and percutaneous cholangiography was compared in 114 consecutive patients with obstructive jaundice. The final diagnosis was obtained by surgery or autopsy. Transhepatic cholangiography diagnosed obstruction and its level in all patients, whereas ultrasonography failed to find obstruction in 3 patients and was unable to determine the level in 11 patients. The cause of obstruction was correctly assessed in 106 patients by transhepatic cholangiography and in 74 by ultrasonography. In 15 patients with obstruction caused by common duct calculi ultrasonography only diagnosed the five, and in 9 patients malignant obstruction was diagnosed as calculi. Ultrasonography is a reliable tool for diagnosis of obstructive jaundice and in most cases for localisation of the level of the obstruction. However, diagnosis of the cause of obstruction and of its precise topography requires direct cholangiography. PMID- 2992013 TI - [Value of sonography for acute diagnosis and for further examination of blunt abdominal trauma in children]. AB - For the investigation of the abdomen following blunt trauma there are, in addition to clinical examination, peritoneal lavage, laparoscopy, sonography, CT and scintigraphy. Sonography has proved particularly reliable without stressing the patient. This has been demonstrated by the examination of 73 children in whom there was only one false positive and no false negative finding. Sonography is valuable not only as a diagnostic method, but is useful for the follow-up of patients with intraperitoneal or retroperitoneal injuries. Particularly in children, a conservative attitude is often justifiable, since recovery frequently follows without surgery (seven out of 30 children in our clinical material, who had sonographic evidence of parenchymal injuries). PMID- 2992014 TI - [Status of penis sonography in andrology]. AB - Good visualization of the anatomic structures of the penis can be achieved by application of high-resolution real-time sonography and a water path. Penis sonography is painless, free of risk and easy to perform. It is ideal for documentation and assessment of pathological alterations of the penis. 80 patients with penile deviation were examined sonographically. In cases of Peyronie's disease, the most frequent clinical picture, it is possible to achieve good localization and measurement of the fibrotic plaques in the area of the tunica albuginea. Calcifications can be clearly detected. In cases of penile deviation associated with the urethral manipulation syndrome, no fibroses are seen in the corpus spongiosum. In 3 patients with erectile impotence after priapism, sonography permitted precise determination of the extent of fibrosis formation in the corpora cavernosa. PMID- 2992015 TI - [Comparative sonographic-anatomic studies aimed at demonstrating intrarenal fine structure]. AB - The detailed structure of the kidneys was studied in 40 excised organs by means of high resolution sonography. It becomes possible to differentiate the details within the renal parenchyma and the sinus. Intrarenal vascular details can also be defined. The clinical relevance of these results is discussed. PMID- 2992016 TI - [Mesentericography: role of intra-arterial DSA in acute occlusive disease]. AB - This study was carried out to evaluate the intra-arterial DSA method for diagnosis of occlusions of the mesenteric vessels. DSA examination in 24 patients (sup. mes. a.n = 24, inf. mes. a.n = 2) demonstrated that image quality was diagnostic in 84%, i.e. contrasting of mesenteric artery including the peripheral branches, when 10 ml of the contrast medium was injected selectively. In 63% of examinations the mesenteric venous return could be documented. Causes of DSA failure were image artefacts due to reduced patient's cooperation and bowel motion. Otherwise the obvious advantage of i.a. DSA was an immediate diagnostic imaging in patients with severe occlusive vascular disease using a manual administration of only small volumes of contrast material. PMID- 2992017 TI - [Embolization of the hepatic artery in primary hepatocellular carcinoma]. AB - Between April 1981 and October 1984, 189 embolisations of the hepatic artery were carried out in 82 patients with primary hepato-cellular carcinomas. Two types of embolisation were performed: 1) embolisation using only gel foam and 2) a combination of peripheral and proximal embolisation using Lipiodol and gel foam. Embolisation of the hepatic artery is indicated for inoperable carcinoma, provided less than 75% of the liver is involved or for localised carcinomas in patients whose general condition makes surgery impossible, often because of severe cirrhosis of the liver. Complications following embolisation include pain, fever and transient changes in liver function. Deaths, abscesses or rupture of the tumour did not occur. PMID- 2992019 TI - [Comparison of x-ray and histological findings in osteosarcomas following preoperative chemotherapy]. AB - Since the introduction of pre-operative chemotherapy, osteosarcomas have shown a more favourable prognosis. Reaction of the tumour due to chemotherapy is judged pre-operative primarily by radiology (plain films, angiography, CT, scintigraphy). There is little evidence concerning the radiological appearances after pre-operative chemotherapy and morphological changes, particularly in respect of tumour regression. Specific radiological changes were therefore compared with pathological findings following chemotherapy and operation in 17 patients with osteosarcomas. Tumours were examined which showed radiological evidence of intra-and extra-osseous sclerosis or lysis and which still were classified as vital tumour tissue. Tumour planes were reconstructed from large histological sections of the operative specimen and compared with the radiological appearances. Sclerosis was found to be due to reactive new bone formation or to mineralisation of the osteosarcomatous tissue. Lysis correlated with persistent vital tumour, or in connective tissue. Nine out of ten cases, regarded as vital on radiological evidence, showed vital tumour cells on histological section. Lyses and scleroses were not reliable indications of the pre-operative state of the osteosarcoma following chemotherapy. On the other hand, combined qualitative radiological criteria for assessing tumour vitality, proved to be helpful. PMID- 2992018 TI - [Spondylolysis and -listhesis. A computed tomographic differential diagnosis of lumbar discopathy]. AB - Spinal CT scans of 680 patients with suspected disc herniation were reviewed to detect lumbar spondylolysis. In this group of patients, 3.23% (n = 22) had pars interarticularis defects at L4 or L5. Disc herniation at the interspace of the pars defect was seen in 27.27% (6/22) of patients with spondylolysis. Using Meyerding's method, which measures the degree of vertebral forward dislocation, grade 2 and grade 3 spondylolisthesis were most often seen. While moderate and severe spondylolisthesis can be already detected via lateral localiser image (scout view), pars defects with only little forward dislocation can be easily underdiagnosed. Sources of diagnostic error are: 1. an atypically posterior disc margin which extends beyond only one vertebral body and presents the appearance of disc herniation, 2. pars defects simulating the adjacent facet joints involve the risk of overlooking spondylolysis, 3. CT sections made through the intervertebral discs and facet joints only, may fail to show up the pars defect 10 or 15 mm above the disc plane. PMID- 2992020 TI - [The Currarino triad. An autosomal-dominant inherited complex of anorectal malformation, sacrococcygeal defect and presacral tumor. Observation of 9 further cases]. AB - Nine cases of a syndrome are described, which is known as the "Currarino triad" and which belongs to the group of malformations in which there is a persistent neurenteric communication. The features of the triad consist of ano-rectal anomalies (particularly ano-rectal stenoses), a curved, but limited, sacro coccygeal defect ("scimitar sacrum") and a presacral tumour, which may be an anterior sacral meningocoele, a teratoma, a cyst (dermoid or neurenteric a cyst (dermoid or neurenteric cyst) or a mixture of these. In at least 50% of cases, the triad is familial and autosomal dominant. Of our nine cases, seven were familial. A dangerous complication is meningitis due to infection of the cystic component, or fistula formation between the colon and spinal canal (one of our cases). Other complications are a fixed filum terminale ("tethered cord") and malformations of the urogenital tract. The risk of malignancy in a teratoma is low, but exists. When considering the indications for surgery, this must be kept in mind, as must the risk of infection, and damage to neural structures during operation. In all cases of early obstipation the sacrum should be x-rayed in order to exclude a Currarino triad. The finding of a sacral defect and a presacral mass makes it essential to obtain CT of the pelvis with contrast in the distal gut and in the meningeal sac. The same is true for screening of the family (including views of the sacrum). PMID- 2992021 TI - [Contrast media in magnetic resonance tomography. A review. 2. Biological basis, research technic and clinical application of gadolinium-DTPA in the diagnosis of intracranial tumors]. AB - The early results of the use of the MR specific contrast medium gadolinium-DTPA in 40 patients with intracranial tumours are reported. Optimal pulse sequences, contrast dose and timing are discussed. The relationship between the blood-brain barrier and contrast distribution within tumour tissue is elaborated. The clinical significance of the application of contrast agents in the diagnosis of brain tumours is evaluated. PMID- 2992023 TI - [The cutting biopsy cannula for histological diagnosis of abdominal and retroperitoneal masses. Sonographic or computed tomographic guided puncture?]. AB - A new cutting biopsy needle has been used to obtain histologically useful material while causing the minimum of trauma. It permits biopsies of organs, but its small external diameter of 0.8 or 0.95 mm makes it possible to carry out transperitoneal puncture of the stomach, colon or liver and of the retroperitoneal space, using a ventral approach. Tissue samples were obtained in 96% of 63 patients. The risk of complications is no higher than for conventional needles used for cytology. The accuracy with which the material can be obtained is therefore the most important problem. The position of the area to be biopsied should determine whether the biopsy is to be aided by sonography or CT. Biopsies of organs can be appropriately carried out under ultrasound control, but processes in the pelvis and in retroperitoneal-paravertebral and extra-peritoneal positions are best biopsied under CT control. PMID- 2992022 TI - [Development of a tissue-equivalent plastic block for the sonographic diagnosis of regions near to the skin]. AB - A very soft and homogeneous plastic material has been developed from polyvinyl chloride with acoustic properties similar to those of human tissue. During an evaluation period of nine months, the material has proved excellent for the sonographic investigation of subcutaneous tissue, even at high frequencies. Significant advantages of the new material as compared with previously available substances depend on its stability, low sound attenuation and absence of reverberations. In order to achieve these desirable properties, twenty plasticisers of different type and concentration were used and the resulting compounds examined for their acoustic properties. PMID- 2992024 TI - [11C-methionine PET, IMP-SPECT, CT and MRI in brain tumors]. AB - 11C-labelled L-methionine uptake was measured in 7 patients with brain tumours prior to surgery, and in 2 patients with infarction, using PET. The strongest uptake occurred in tumours with a high grade of malignancy (astrocytoma IV: Tumour/Non-Tumour T/NT = 2.6) while low-grade tumours accumulated less activity (astrocytoma II: T/NT = 1.4). Conventional 99mTc DTPA scans revealed a damage of blood brain barrier (BBB) in 4 patients (2 infarctions) with no or only slight 11C-methionine accumulation, while one patient with negative 99mTc-scan and negative CT accumulated methionine in the tumour region (astrocytoma II). PET, MRI, and CT are complementary with regard to extent of tumour tissue, necrotic areas and oedema. None of the brain tumours or infarcted regions took up IMP (123I amphetamine). The investigated regions showed definite uptake defects ranging from 10-50% if compared with the contralateral side. In view of the chemical pathway of methionine it is concluded that the uptake reflects metabolic activity in brain tumour tissue rather than a diffuse uptake due to BBB damage. PMID- 2992025 TI - [NMR studies of the central nervous system in pediatrics]. AB - The central nervous system in 70 infants, children and adolescents has been examined by NMR. Phase I of the continuing investigation dealt with the applicability and scope in this group of patients. A retrospective comparison with CT was used to examine the possibility of enhancing and differentiating various tissues. The results have shown that NMR can extend the diagnostic possibilities beyond those of CT in the paediatric age group. PMID- 2992026 TI - [Central traumatic cerebral hemorrhage in the computed tomogram]. AB - Traumatically caused central brain haemorrhages are relatively rare. From 1976 through 1984 3598 patients were submitted to computed tomography examination in our department after craniocerebral trauma. Central lesions could only be found in about 3% of the injured. They generally occurred in combination with other severe damage of the skull and skull contents. Most frequently, such central haemorrhages could be found in the basal ganglia occasionally extending into the adjacent medullary layer resulting in large intracerebral haematoma. The right hemisphere was affected significantly more frequently and more severely by such large-size haemorrhages than the left hemisphere. The thalamus region ranking second in localisation of central traumatic haemorrhages was virtually never found to be the origin of large haematomas. In general, no definite distinction could be made between primary and secondary traumatic haemorrhages. We identified, however, a few cases of purely central bleeding without accompanying brain lesions. These haematomas, which were most probably caused primarily by trauma, as well as those with associated damage, preferred the regions of basal ganglia and thalamus. The prognosis of central brain haemorrhages was relatively poor with a 42% lethality rate. However, it depended on the severity of the accompanying brain lesions. Thus, isolated central haemorrhages even had a markedly favourable prognosis. The number of survivors of central bleeding turned out to be approximately the same as the number of deaths, the ratio thus being 1:1. Nevertheless, we think that especially small lesions occur more frequently and have a better survival rate than had been supposed up to now. PMID- 2992027 TI - [High-resolution computed tomography in primary and secondary lesions of the optic nerve sheath complex ]. AB - High resolution CT was found to be an extremely useful and sensitive method for evaluation and diagnosis of lesions involving the optic nerve sheath complex. Segmental "tram tracking" associated with fusiform or focal eccentric nerve sheath enlargement was confirmed to be of neoplastic origin in all cases, whereas "tram tracking" and tubular nerve sheath thickening included tumorous and a great variety of non-tumorous lesions as well. PMID- 2992028 TI - [Technic and results of spinal computed tomography in the diagnosis of cervical disk disease]. AB - We give a description of a technique of the patient's positioning with traction of the arms during the cervical spinal computed tomography which allows to draw the shoulders downwards by about one to three cervical segments. By this method the quality of the images can be improved in 96% in the cervical segment 6/7 and in 81% in the cervical/thoracal segment 7/1 to such a degree that a reliable judgement of the soft parts in the spinal canal becomes possible. The diagnostic reliability of the computed tomography of the cervical disc herniation is thus improved so that the necessity of a myelography is decreasing. The results of 396 cervical spinal computed tomographies are presented. PMID- 2992029 TI - [Differential diagnosis of cystic and cystic-like lesions of the jaw]. AB - The difficulties in the differential diagnosis of jaw cysts are demonstrated by 10 selected case studies. Panorama radiographies are basic for radiodiagnosis. Supplementary radiographs, including CT, are often necessary. In general, the final diagnosis can be given after synopsis of clinical, radiographic and patho histological findings. PMID- 2992030 TI - [MR of the knee joint. Imaging of the normal anatomy and pathological findings using surface coils]. AB - A special surface coil was developed with the intention of producing optimal resolution and signal-to-noise ratio for the examination of the knee joint by MRI. Eight volunteers and 25 patients with various abnormalities of the knee have been examined. Normal anatomical structures can be differentiated and there is high diagnostic reliability for lesions of the meniscus, as confirmed by arthroscopy and surgery. Cartilage lesions are less well demonstrated. Total rupture of cruciate ligaments is clearly shown, but partial rupture can often be recognised on the basis of indirect signs only. Good diagnostic results were obtained in cases of osteochondritis dissecans and chondropathia of the patella. PMID- 2992031 TI - [Imaging of the Achilles tendon on the computed tomogram. Normal findings and pathological changes]. AB - Ligaments and tendons, including the Achilles tendon, show the highest density among normal soft tissue structures in the body. Traumatic and degenerative changes of the Achilles tendon are often associated with marked thickening and reduction in density associated with increased opacity of the space in front of the Achilles tendon. These changes are easily demonstrated by CT, whereas conventional radiological techniques only show non-specific changes. Twenty-five patients were examined, including nine with pain, seven following rupture of the Achilles tendon and nine post-operative controls; it was found that CT can add information important for the diagnosis and treatment planning of abnormalities of the Achilles tendon. PMID- 2992033 TI - Cerebral gigantism (Sotos' syndrome). Metacarpophalangeal pattern profiles. AB - The metacarpophalangeal pattern (MCPP) profile in 17 out of 22 patients with Sotos' syndrome was studied throughout the growth period, from 2 weeks to 19 years of age. Two typical MCPP profiles will be described. The first type of profile was found in 13 patients. Although the form of the profile is the same throughout the years, the lengthening of the fingers is most pronounced around 2 years of age. Before and after that period there is a flattening of the curve. The metacarpals are relatively longer below the age of 3 years, above 3 years the lengthening of the proximal phalanges is more pronounced in the profile. The second type of profile was found in 4 patients. This curve is almost a mirror image of the first profile. The fingers are not as long as in the first type and the profile is less pronounced. In one of the 17 patients the transition from the first type to the second type could be demonstrated. One patient had a normal profile. PMID- 2992032 TI - [Separate measurements of compact and spongy bone density using a transverse rotation scanner. Determination of mineral content of the radius using a least squares algorithm]. AB - The absorption profile of the distal forearm, which depends on bone mineralisation, was measured using a high resolution microcomputer-controlled transverse rotational scanner with a tightly collimated I-125 photon source and a scintillation counter. Reconstruction of the anatomical structures within the measured area occurred in two stages. Subsequently the linear attenuation coefficient for soft tissues, compacta and spongiosa was obtained by a "least square" algorithm. The absorption coefficient of the compacta and spongiosa provided a measure of bone mineralisation. Correlation between the reconstructed images and the calculated results and measurements was tested by calculation. Ten measurements of a model and of in vivo bones resulted in a coefficient of variation of 1.8% and 3.3% for the compacta and 2.5% and 3.8% for the spongiosa. Relative deviation from the standard varied from +1.3% to -1.0%, depending on mineral content. This new, technically simple procedure for the separate determination of the compacta and spongiosa in the distal radius is useful for the early recognition of changes in bone mineralisation which are usually apparent first in the spongiosa. PMID- 2992034 TI - [A simple method for imaging disorders of cardiac wall contraction using MRT]. AB - By means of ECG triggering, it is possible to demonstrate various phases of cardiac action. It is then possible to derive a measure for cardiac function. The disadvantage of the method consists of the long measurement period required and the creation of movement artifacts. A method is discussed which would permit the demonstration of normal cardiac movement during a short time span and which reduces artifacts. In order to obtain optimum results, a new sequence of measurements is suggested. This is based on spin-echo sequences and can readily be used with any type of MRT apparatus. PMID- 2992035 TI - [Computed tomography of esophageal carcinoma. Correlation between computed tomographic and postoperative findings]. AB - Between March, 1980 and January, 1984, computerized tomography (CT) was performed on 110 patients with proven esophageal carcinoma. In 26 patients, information obtained preoperatively by CT was compared with results of intraoperative exploration or histologic examination of resection specimen. Correlation analysis showed that accuracy of CT in assessing actual tumor size and mediastinal or abdominal lymph node involvement is rather limited, while correct results were obtained in between 84 and 100 per cent of patients as far as identification of invasion of adjacent organs is concerned. We thus advocate routine use of CT in the process of preoperative assessment of operability and staging. PMID- 2992036 TI - Herniography of femoral, obturator and perineal hernias. AB - Positive contrast herniography was used in the workup of 550 patients with unclear groin pain. The majority of these patients had rather characteristic hernias of indirect, direct or femoral type. However, now and then diagnostic problems arose. A femoral hernia may look like a direct, indirect or even obturator hernia. There is also a variety of multilocular femoral hernias and other types. A femoral hernia may be present together with other hernias in the ipsilateral or contralateral groin. Obturator hernias are usually small but are always confined to the obturator canal laterally in the obturator foramen. Abnormalities in the pouch of Douglas may include a deep rectogenital pouch, diverticula and true herniations. These uncommon herniographic findings are described and discussed. PMID- 2992037 TI - [Initial experiences with MR in liver tumors]. AB - We examined 21 patients with focal lesions of the liver. Routinely we used T1 weighted, proton weighted and T2 weighted measurement modes, mainly with repetition times of 1.6 sec and echo delay times of 35 or 120 msec. Using these parameters we can see characteristic changes of the signals of the liver tumours. Cystic lesions usually show a strong decrease of the signal in the T1 weighted images in comparison with the normal liver pattern, in the proton weighted images a weak decrease but also in some cases a weak increase of the signal; in the T2 weighted images they show signals of very great intensity. We can differentiate haemangioma of cystic lesions because of the very strong signal in the proton weighted images in comparison with the normal liver pattern, which we could not see in any other focal liver disease. Metastases and hepatoma produced low signal intensity in the T1 weighted image. The proton weighted and the T2 weighted images show signals with a slightly greater intensity compared with the normal pattern, i.e. a very good possibility to distinguish hepatoma and metastases from cystic lesions or haemangioma. The differentiation from hepatoma and metastases cannot be made with NMR up to now. We are also not able to differentiate the focal nodular hyperplasia (FNH) from metastases. We used a 0.35 T supraconductive magnetic system. PMID- 2992038 TI - [Nuclear spin tomography in breast diagnosis]. AB - The authors report about their preliminary experience with MRI in the diagnosis of breast disease. 50 breast masses in 41 consecutive patients have been evaluated by MRI and mammography, some of which have been evaluated by ultrasound, as well. All masses have consequently been biopsied. They include 32 carcinomas, 1 secondary malignant lymphoma, 4 fibroadenomas, 2 papillomas, 3 cysts, 1 hamartoma and 5 dysplastic nodules. Advantages and disadvantages of MRI of the breast are discussed. Possible future indications are suggested for selected cases. PMID- 2992039 TI - [Circumscribed and diffuse skin thickening (peau d'orange) of the female breast]. AB - Skin thickening of the breast observed by mammography should always alert the radiologist to suggest the possibility of carcinoma. The roentgenographic appearance of skin thickening, however, is nonspecific and identical in each disorder, and provides no clue to its benign or malignant origin. It is therefore important to the radiologist to be familiar with the possible etiologies, which can often be simply evaluated by obtaining an appropriate medical history, clinical inspection or a thorough physical examination of the breast. A review of the mammary skin thickening is given, and the differential diagnosis is discussed with reference to the literature. PMID- 2992040 TI - [Computed tomography and nuclear spin tomography in peripheral lymphedema]. AB - The first reported findings on CT and N.M.R. in two cases of idiopathic peripheral lymphoedema are described. These methods have compared with the generally available volume estimations (volume estimation according to Kuhnke, immersion plethysmography) and have been considered in relation to visual and lymphographic examinations. PMID- 2992041 TI - [Random sample size in ROC analyses]. AB - ROC analysis has proved useful in assessing diagnostic efficiency. Large x-ray film series of thoracic images intended for cardiovascular diagnosis were used for studying diagnostic efficiency in relation to random sample size. Exploration of random samples from two classes of findings requiring diagnostic differentiation, with the same number of x-ray films in each class, showed satisfactory convergence between the radiologists' rating and the ROC curve if the size of the random sample groups was about 200 images each or larger. The smaller the random sample size (less than 200 images), the greater the scatter of the rating performance. In another series the rating ability of experienced radiologists was compared with that of a semi-automatic classificator. The semi automatic classificator attained almost the same performance index as the low performance evaluator. PMID- 2992042 TI - Legionella pneumonia in children: radiological findings in a 3-year-old child. PMID- 2992043 TI - [Extensive Hand-Schuller-Christian disease (histiocytosis X)]. PMID- 2992044 TI - [Radiographic aspects of a large splenic artery aneurysm--gastrointestinal radiography and computed tomography]. PMID- 2992045 TI - [Spontaneous rupture of an intercostal artery aneurysm]. PMID- 2992046 TI - Fibromuscular dysplasia of the ulnar artery. PMID- 2992047 TI - [Giant aneurysm of the intracranial internal carotid artery. Methods of radiological diagnosis]. PMID- 2992048 TI - [Hematogenous metastasis caused by angiography?]. PMID- 2992049 TI - [Familial congenital diaphragmatic defect]. PMID- 2992050 TI - Episternal ossicles: a normal CT variant. PMID- 2992051 TI - [Extraosseous nuclide uptake during skeletal scintigraphy. Heparin-calcium induced 99mTc-HMDP accumulation]. PMID- 2992052 TI - EEG changes induced by convulsants interfering with GABAergic inhibition in rats. I. Cortical recordings. AB - Electrocorticographic changes induced by 3 convulsant drugs interfering with the GABA system were studied in 73 adult rats. Aminooxyacetic acid first elicited changes (periods of theta waves) nearly constantly in occipital regions. Bicuculline induced not only occipital theta waves but also groups of spikes with the same frequency in frontal regions; there was no predominance of one of these regions. 3-Mercaptopropionic acid invariably elicited groups of spikes in the frontal region as the first sign of its action. As it progressed, this rhythmic activity became generalized so that differences among the 3 drugs disappeared. Paroxysmal activity induced by the 3 drugs studied was identical. It always started with rhythmic spikes which were gradually transformed into spike-and-wave rhythm. Differences among the 3 convulsants studied were found in the very first electrocorticographic changes. Long latency to the onset of action of 3 mercaptopropionic acid and especially of aminoocyacetic acid qualifies these drugs for studies of preparoxysmal changes. PMID- 2992053 TI - [Embolization of the biliary tract caused by necrotic hepatocarcinoma, an unusual cause of obstructive jaundice (apropos of 2 cases)]. PMID- 2992054 TI - [A new case of fibrolamellar hepatocarcinoma]. PMID- 2992055 TI - [Converting enzyme inhibitors in the therapy of primary arterial hypertension]. PMID- 2992056 TI - [Drugs that block the angiotensin converting enzyme]. PMID- 2992057 TI - Basic principles in imaging of regional cerebral metabolic rates. PMID- 2992058 TI - The interaction of eptazocine, a novel analgesic, with opioid receptors. AB - The profile of action of eptazocine, a novel analgesic, on opioid receptors was investigated. Eptazocine caused a concentration-dependent inhibition against the [3H]-naloxone [( 3H]-NLX) specific binding to rat brain synaptic membrane in the absence of sodium cation and GTP (IC50; 7.83 +/- 1.57 microM). The ratios of IC50 values between the absence to the presence of sodium cation alone or sodium cation and GTP were 3.89 and 4.35, respectively. In addition, eptazocine (10 microM) also produced the significant decrease of [3H]-NLX specific binding in the mouse brain synaptic membrane. Moreover, the same dose eptazocine significantly decreased the [3H]-ethylketocyclazocine [( 3H]EKC) specific binding, but not [3H]-phencyclidine [( 3H]-PCP). These results suggest that eptazocine interacts with opioid receptor, and is classified as one of the opiate agonist-antagonist analgesics. PMID- 2992059 TI - [Peripheral sensory neuropathy produced by a megadose of vitamin B6]. AB - Clinical cases of sensory neuropathy produced by a megadose of vitamin B6 have been reported in English literatures. We investigated the ordinary daily dosage and maximal dose of vitamin B6 widely adopted in Japan, and the amount of vitamin B6 per unit (per tablet, capsule or ampule) available in our medical practice. We concluded that in Japan it is very rare to administer such a large dose of vitamin B6 that produced sensory neuropathy described in the literatures. In our experimental study, Sprague-Dawley rats were intraperitoneally given a total amount of 14,000 mg/kg of body weight of pyridoxine hydrochloride in ten separate doses. They developed an ataxic gait. The occurrence of the degeneration of nerve cell bodies and peripheral axons of lumbar primary sensory neurons were histologically demonstrated. Although in Japan no clinical cases of neuropathy produced by a megadose of vitamin B6 have been reported to our knowledge, it is necessary to be aware of the possible occurrence of such neuropathy among patients with polyneuropathy of unknown etiology or who have been receiving vitamin B6 for a long time. PMID- 2992060 TI - [The non-medicinal therapeutic arsenal in essential arterial hypertension]. PMID- 2992061 TI - Immunology of viral diabetes. PMID- 2992062 TI - Role of papilloma virus in proliferative squamous lesions. AB - HPVs are associated with a variety of proliferative squamous lesions. 27 different types of HPV have been identified by DNA molecular hybridization studies. Genus- and type-specific HPV structural antigens can be detected in approximately half of benign warts and condylomata by immunocytochemistry; positive lesions are considered infectious. Genus- and type-specific putative HPV DNA sequences replicating as episomes can be recovered from the majority of benign (exophytic and flat condylomata) and malignant squamous lesions of the cervix and anogenital area. The type of HPV determines, in part, the anatomic site, clinical appearance, and natural history (including potential malignant conversion) of the lesion. PMID- 2992063 TI - [Which therapeutic attitude should be adopted in the presence of non-small cell bronchopulmonary cancer?]. AB - Complete surgical excision is the best method of obtaining cure. However, this is only possible in less than one quarter of the patients. In all of the other patients, the hopes for cure are based on combinations of chemotherapy radiotherapy-surgery, in particular preoperative chemotherapy. Phase II trials of chemotherapy are underway. Real progress will only be achieved when joint multicentric studies can be conducted. PMID- 2992064 TI - [Trophoblastic hyperplasia and placental carcinomas]. AB - The mechanism instigating triploidy is linked to placental aspects. When 2 of 3 gametes are paternal in origin, molar pregnancy results, while if 2 of 3 gametes are of maternal origin the placenta is normal (15% of cases). Hydatidiform mole and chorioadenoma destruens or invasive mole are indistinguishable one from the other in terms of clinical appearance, histology and course of disease, and are always benign. Their karyotype is 46,XX (except for 3% which are 46,XY) completely paternal in origin and usually homozygotes. Trophoblastic carcinoma however is aneuploid and heterozygotic which emphasizes its allograft nature i.e. with no apparent relationship to hydatidiform mole. PMID- 2992065 TI - [Papillomavirus, herpes virus and vulvar cancer]. AB - Vulvar carcinoma is relatively rare but genital warts (condylomas) are very frequent findings. Very old diseases, the roles of certain viruses in their pathophysiology has been shown. The mechanism of viral infestation is reviewed. Absolute proof of precise viral oncogenic roles in humans remains unrealized despite definite clinical and epidemiologic arguments; the role is that of a cocarcinogen acting in conjunction with physical, chemical, clinical and immunological factors in an as yet obscure chronological order. For the moment, viral vulvar lesions must be considered as cancer risks and be treated as such. PMID- 2992066 TI - [After-care centers]. PMID- 2992067 TI - The preparation of bioactive 125I-gastrin, using Iodo-gen as oxidizing agent, and the use of this tracer in receptor studies. AB - The preparation of 125I-labelled gastrin 1-17, using Iodo-gen as oxidizing agent, is described. Mono-125I-gastrin was purified from the iodination mixture by gel chromatography, followed by ion-exchange chromatography. The Iodo-gen-labelled 125I-gastrin tracer showed a slightly higher immunoreactivity than a similarly purified tracer produced by a gentle chloramine-T method (50% binding of 0.45 fmol tracer at an antibody dilution of 945,000 versus 780,000, respectively). The Iodo-gen-labelled 125I-gastrin, which had the same biological activity as native gastrin when tested in conscious gastric fistula rats, was tested for specific binding at 30 degrees C in pronase-isolated rat gastric fundic cells and plasma membranes from the same area. A specific binding with a Kd of 4.5 X 10(-9) M was found to isolated fundic cells. This binding was rapid and reached equilibrium within 40 min. The dissociation induced by a 10-fold dilution of the incubation medium was biphasic, with a rapid initial phase and a slow late phase, indicating two different binding sites. On the other hand, no reproducible specific binding of the tracer to plasma membranes was obtained. This study shows that Iodo-gen is suitable as an oxidizing agent in iodination of gastrin without loss of biological activity. The tracer produced may be used in receptor studies for isolated cells, whereas the use of plasma membranes for studying gastrin receptors needs reevaluation. PMID- 2992068 TI - Acid secretion in isolated gastric glands from healthy subjects and ulcer patients. AB - Isolated human gastric glands provide an in vitro model that can yield significant information about the mechanisms regulating gastric acid secretion at the parietal cell level. Aminopyrine, a weak base that accumulates in acid compartments, has been used as an indirect probe of H+ secretion. By means of a microscale technique it was possible to isolate oxyntic glands from gastroscopic biopsy specimens and thereby enable studies of healthy subjects and non-operated ulcer patients. Histamine (5.4 X 10(-5) M) and db-cAMP (10(-3) M) both induced a pronounced response, whereas the response to carbachol (4.5 X 10(-6) M), although still statistically significant, was less potent. The response to stimuli was twice as high in duodenal ulcer patients as in normal individuals. In contrast, the response in patients with a gastric ulcer located either in the prepyloric region or at the minor curvature on the antrum-corpus border was of the same magnitude as in healthy subjects. Pentagastrin did not induce any response in isolated gastric glands from normal individuals. Gastric acid secretion in vitro, measured as aminopyrine accumulation, did not decrease with increasing age of the individuals. PMID- 2992069 TI - Dietary aspects of peptic ulcer disease. AB - Diets modify gastric function in various ways. Fibre influences acidity, pepsin and bile acid concentrations both in vitro and in vivo. Both animal studies and clinical studies in humans give reason to believe that a diet rich in fibre may protect against the development of peptic ulceration. PMID- 2992070 TI - Autologous mixed lymphocyte reaction in man. XI. Deficiency of autologous mixed lymphocyte reaction and abnormalities of monoclonal antibody-defined T-cell subsets in chronic mucocutaneous candidiasis. AB - The autologous mixed lymphocyte reaction (AMLR) and T-cell subsets defined with monoclonal antibodies were examined in the peripheral blood of six patients with chronic mucocutaneous candidiasis (CMC). The AMLR was deficient in four of six patients when compared with simultaneously studied healthy controls. These patients had either overt endocrinopathy or circulating autoantibodies. In two of three patients with low AMLR, after the depletion of OKT8+ T cells no enhancement in the AMLR was observed, demonstrating that the deficiency of the AMLR was not due to increased suppressor OKT8+ T-cell activity. However, the third patient demonstrated almost complete reconstitution of the AMLR response after such depletion, suggesting that OKT8+ suppressor T cells were responsible for decreased AMLR in this patient. Two patients had a low ratio of OKT4/OKT8 phenotype T cells. T cells with Tac antigen (that is, present on activated T cells) were increased in four of six patients. No correlation was observed between the deficiency of the AMLR and the proportions of T-cell subsets. This study demonstrates a deficiency of the AMLR in some patients with CMC which is associated with increased OKT8+ suppressor T-cell activity or with the functional deficiency of responder OKT4+ T cells. PMID- 2992071 TI - Isolation of highly purified lymphocyte subsets for functional studies by means of an indirect rosette technique. AB - An indirect rosette assay, utilizing ox erythrocytes (RBC) coupled with rabbit anti-mouse IgG and lymphocytes sensitized with monoclonal mouse antibodies against membrane markers, was used for purification of lymphocyte subsets that were functionally intact. Either peripheral blood mononuclear cells (PBMC) or T lymphocytes isolated by sheep RBC rosetting could be used as starting material for obtaining pure T-cell subsets (T4 or T8). The following steps of the method were evaluated: the procedure of coupling rabbit anti-mouse IgG to ox RBC via the CrCl3 method, the experimental conditions for specific rosetting, and the use of Percoll for the separation of rosettes from the non-rosetting cells. Under optimal experimental conditions the recovery of positively selected cells was 45 55% of the cells originally present in the PBMC. The purity of these cells reached a value of more than 95%, whereas the contamination of the depleted fraction was less than 3%. The functional integrity, manifesting itself as proliferation after mitogen stimulation and as regulatory influences on in vitro Ig synthesis, appeared to be unimpaired. The described technique may be applied to the purification of various cell subpopulations for functional studies, provided monoclonal antibodies against membrane antigens are available. PMID- 2992072 TI - HTLV in Sweden: antibodies to HTLV I antigens in experimental monkeys and their caretakers. AB - 252 monkeys kept at 4 different Swedish universities and laboratories for experimentation were screened for antibodies to HTLV I associated antigens by means of a sensitive membrane antigen enzyme immunoassay (MA-ELISA). 17/185 Macaca fascicularis, 1/56 M. mulatta, 0/1 Cercopithecus aetiops and 0/10 Saimirii squiureus had antibodies. All of 11 MA-ELISA positive animals which were subjected to further testing were also positive in a competition assay for anti HTLV p19 antibodies and in an anti-whole virion enzyme immunoassay. One colony of 32 M. fascicularis monkeys from the Philippines contained 7 antibody-positive animals. Except for one M. fascicularis which suffered from a chronic dermal lesion, major disease was not observed in any of the antibody-positive animals. None of 28 animal caretakers or experimenters, of which several had been repeatedly exposed to blood from antibody-positive animals, had antibodies measurable by the MA-ELISA. The contagiosity for humans of the majority of the antibody-positive monkeys thus appears to be relatively low. We conclude that the presence in Sweden of HTLV I antibody-positive animals probably does not constitute a great health risk. However, we consider it appropriate that antibody positive animals should be handled with special care. PMID- 2992073 TI - Cytomegalovirus mononucleosis in children and adults: differences in clinical presentation. AB - In 2 previously healthy groups of 14 children and 17 adults with cytomegalovirus mononucleosis, significant clinical differences were observed. Cervical lymphadenopathy, hepatomegaly and lymphocytosis (greater than 5000/microliter) were more common in children and protracted fever more common in adults. Exudative tonsillitis indistinguishable from infectious mononucleosis was sometimes seen in children but never in adults. PMID- 2992074 TI - Intravenous foscarnet for the treatment of severe cytomegalovirus infection in allograft recipients. AB - Foscarnet, trisodium phosphonoformate, was administered intravenously to 6 immunosuppressed patients with life-threatening cytomegalovirus infection. Three of the patients were recipients of a kidney and 3 of a bone-marrow transplant. Favourable clinical responses were seen in 5 of the patients, 2 of whom were still in good health 5 and 8 months after the infection had cleared up. No toxic effect of the drug was detected. The results seem to justify further trials, in which foscarnet should be introduced at an earlier stage of the disease. PMID- 2992075 TI - Automated cytochemical differential leucocyte count in patients hospitalized with acute bacterial infections. AB - Absolute blood levels of various leucocyte types were measured by an automated cytochemical differential count (Hemalog D/90) in 84 adult patients hospitalised with an acute bacterial infection. The neutrophil granulocyte levels varied widely in all patient categories ranging from a normal value to a high count. Neutrophilic granulocytosis defined by a count exceeding 6.0 X 10(9)/l was detected in 47 patients (56%). An increased level of neutrophils with a high myeloperoxidase activity (Hpx cells) representing leucocytes recruited from the bone marrow was found in 51 patients (61%). Monocytosis was found in 25 (30%) and lymphocytopenia in 23 patients (27%). A high number of large unstained cells indicating an increase in atypical lymphocytes was detected in only 3 persons. Entirely normal cytochemical parameters were recorded in 13 patients (15%). The relationship between various leucocyte types was investigated in 26 patients with erysipelas. A negative association (r = -0.59) was observed between the neutrophil and the monocyte count. There was no correlation between the neutrophil and the lymphocyte count, nor between the levels of monocytes and lymphocytes. A positive correlation (r = 0.76) was established between the neutrophil count and the level of Hpx cells indicating that recruitment of immature cells from the bone marrow reserve pool is a feature of patients with erysipelas. The high precision and reproducibility of the cytochemical differential count indicate that the variability noted in the disease categories studied is a biological one, being the result of the complex interaction of invading microorganisms with the host defence system. PMID- 2992077 TI - Infiltrating renal collecting duct carcinoma associated with epithelial dysplasia of the renal pelvis. AB - A case of multifocal urothelial dysplasia in the renal pelvis and simultaneously occurring dysplasia in collecting ducts with invasion in the renal medulla is presented. The essential role of cytopathology in diagnosis is emphasized. PMID- 2992076 TI - Deterioration of renal function during angiotensin converting enzyme inhibition in hypertensive patients with a poorly functioning solitary kidney. AB - During treatment with the angiotensin converting enzyme inhibitor, Captopril, glomerular filtration rate (GFR) decreased in three unilaterally nephrectomized hypertensive patients with a poorly functioning remaining kidney. The fall in GFR was not related to changes in the blood pressure, and was reversed when Captopril was stopped. In a fourth nephrectomized patient the initially normal GFR was not affected by captopril. These observations point to a functional reduction in GFR due to a withdrawal of an intrarenal action of angiotensin II. Maintenance of GFR may become critically dependent on a functioning renin-angiotensin system when renal perfusion pressure is reduced to a certain degree. Inhibition of angiotensin II formation may lead to a decrease in GFR in patients with renal hypertension and pre-existing renal insufficiency. PMID- 2992078 TI - Ultrasound diagnosis of carcinoma of the urachus. AB - In a 48-year-old man with dysuria and microscopic haematuria, pyelography, cystoscopy and bimanual bladder palpation showed no definite abnormality. Because of continued symptoms, ultrasound scanning of the abdomen was done three months later. The examination revealed a tumour in close relation to the top of the bladder. Malignant cells were aspirated at ultrasonically guided puncture, and a urachus tumour was suspected. For early diagnosis of urachus tumours, ultrasound investigation of vague bladder symptoms is required. PMID- 2992079 TI - The occurrence of copper, iron, zinc and other elements and the nature of some copper and iron complexes in humic substances from municipal refuse disposed of in a landfill. AB - Municipal refuse was allowed to decompose in a simulated landfill for 20 months. Three different models were studied in which the refuse, in 40 m3 lots, was either compacted or mixed 2:1 with sewage sludge, the latter being studied in both uncompacted and compacted states. At 2, 6, 12 and 20 months, humic substances were extracted from samples with 0.1 M Na2P4O7 and 0.1 M NaOH, and humic acids isolated by precipitation after acidification with HCl. The humic substances were examined by electron paramagnetic resonance (EPR) to determine the free radical content and the nature of some of the metal complexes present. The principal form of copper was present in either a square planar or a tetragonally-distorted octahedral environment, probably coordinated to two nitrogen and two oxygen atoms. Fe3+ was present in at least three different environments. In one, it was probably in a complex with rhombic symmetry; another showed Fe3+ in an axially-symmetric environment, most likely as a ferric porphyrin. No distinctions could be made between the concentrations or forms of metals present in the refuse humic substances as a result of adding sewage sludge, but the additions increased the yield of humic substances, particularly in the uncompacted landfill. Humic substances in the refuse retain metal elements in complexed forms which will restrict their release from the landfill. PMID- 2992080 TI - Effects of genomic position on the expression of transduced copies of the white gene of Drosophila. AB - The white gene of Drosophila is expressed normally when introduced at many different sites in the genome by P-element-mediated DNA transformation, but is expressed abnormally when inserted at two particular genomic positions. It is now demonstrated that the mutant expression in these two cases is caused by the surrounding chromosomal region into which the white gene has been inserted. The white gene could be moved from these two positions, where it confers a mutant phenotype, to other positions in the genome where it confers a wild-type phenotype. However, flies in which white has been moved to one new location have an unusual mosaic phenotype. PMID- 2992081 TI - Infection of HTLV-III/LAV in HTLV-I-carrying cells MT-2 and MT-4 and application in a plaque assay. AB - The human T-cell lines MT-2 and MT-4 carry the human T-cell leukemia virus type I (HTLV-I). When MT-2 and MT-4 were infected with HTLV-III, the probable etiologic agent of the acquired immune deficiency syndrome (AIDS), rapid cytopathogenic effects and cytotoxicity were observed that made it possible to titrate the biologically active virus in a plaque-forming assay. The cytopathogenic effects were preceded by the rapid induction and increase of HTLV-III antigens as revealed by immunofluorescence and immunoprecipitation. Activities of HTLV-III were neutralized by the human antibodies against the virus when immunofluorescence and plaque assays were used. Essentially the same results were obtained with the lymphadenopathy-associated virus (LAV1). PMID- 2992082 TI - The pX protein of HTLV-I is a transcriptional activator of its long terminal repeats. AB - Expression of the pX protein of human T-cell leukemia virus type I (HTLV-I) in animal cells demonstrates that this protein is a specific transcriptional activator of the long terminal repeats (LTR) of HTLV-I. Several other promoters are not affected by pX. No lymphocyte-specific factors are required for this activation. pX can be detected in the nucleus of transfected monkey kidney cells (line CV1) by indirect immunofluorescence. These results indicate that the pX protein is essential for the replication cycle of the virus and that it may be directly involved in the immortalization of human lymphocytes by HTLV-I. PMID- 2992083 TI - Origin of human small cell lung cancer. PMID- 2992084 TI - Genomic diversity of human T-lymphotropic virus type III (HTLV-III). AB - The DNA genomes of human T-lymphotropic virus type III (HTLV-III) isolated from 18 individuals with AIDS or who were at risk for AIDS were evaluated for evidence of variation. Although all of the 18 viral DNA's hybridized throughout their entire genomes to a full-length cloned probe of the original HTLV-III isolate, each of the 18 isolates showed a different restriction enzyme pattern. The number of restriction site differences between isolates ranged from only 1 site in 23 to at least 16 sites in 31. No particular viral genotype was associated with a particular disease state and 2 of the 18 patients had evidence of concurrent infection by more than one viral genotype. Propagation of three different viral isolates in vitro for up to 9 months did not lead to detectable changes in their restriction patterns. These findings indicate that different isolates of HTLV-III comprise a spectrum of highly related but distinguishable viruses and have important implications regarding the pathogenicity of HTLV-III and attempts to develop effective diagnostic, therapeutic, and preventive measures for this virus. PMID- 2992085 TI - Regeneration of functional synapses between individual recognizable neurons in the lamprey spinal cord. AB - In 4- to 5-year-old sea lamprey larvae that had recovered from complete transection of the spinal cord, pairs of giant interneurons on opposite sides of the scar were impaled with microelectrodes. In 4 of 30 pairs, stimulation of the caudal cell elicited a monosynaptic electrochemical excitatory postsynaptic potential in the rostral cell. Fifty percent of such pairs were synaptically linked in control lampreys without transections. These results show regeneration of functional synaptic connections between individual neurons in a vertebrate central nervous system. PMID- 2992086 TI - Huntington's disease: two families with differing clinical features show linkage to the G8 probe. AB - To test the hypothesis that interfamily variability in Huntington's Disease (HD) is due to mutation at different loci, linkage analysis was undertaken in two large HD kindreds that differed in ethnicity, age-at-onset, and neurologic and psychiatric features. Both families showed linkage of the HD locus to the G8 probe. Several recombinants were documented in each family, and the best estimate of the recombination fraction for the two families was 6 percent with a 95 percent confidence interval of 0 to 12 percent. Although the data support the existence of a single HD locus, use of the G8 probe for presymptomatic testing in these kindreds would have resulted in a 12 percent error rate in genotype assignment at the HD locus. PMID- 2992087 TI - A crystalline view of protein-DNA binding. PMID- 2992088 TI - Infection of the basal ganglia by a murine coronavirus. AB - The coronavirus, mouse hepatitis virus strain A59 (MHV-A59), causes mild encephalitis and chronic demyelination. Immunohistochemical techniques showed that MHV-A59-infected C57BL/6 mice contained dense deposits of viral antigen in the subthalamic nucleus and substantia nigra, with fewer signs of infection in other regions of the brain. The animals showed extra- and intracellular vacuolation, neuronal loss, and gliosis in the subthalamic-nigral region. Such localization is unprecedented among known viral encephalitides of humans and other species. This infection by a member of a viral class capable of causing both encephalitis and persistent infection in several species may be related to postencephalitic parkinsonism. PMID- 2992089 TI - Amplification of a novel v-erbB-related gene in a human mammary carcinoma. AB - The cellular gene encoding the receptor for epidermal growth factor (EGF) has considerable homology to the oncogene of avian erythroblastosis virus. In a human mammary carcinoma, a DNA sequence was identified that is related to v-erbB but amplified in a manner that appeared to distinguish it from the gene for the EGF receptor. Molecular cloning of this DNA segment and nucleotide sequence analysis revealed the presence of two putative exons in a DNA segment whose predicted amino acid sequence was closely related to, but different from, the corresponding sequence of the erbB/EGF receptor. Moreover, this DNA segment identified a 5 kilobase transcript distinct from the transcripts of the EGF receptor gene. Thus, a new member of the tyrosine kinase proto-oncogene family has been identified on the basis of its amplification in a human mammary carcinoma. PMID- 2992090 TI - The neu gene: an erbB-homologous gene distinct from and unlinked to the gene encoding the EGF receptor. AB - The neu oncogene, identified in ethylnitrosourea-induced rat neuroglioblastomas, had strong homology with the erbB gene that encodes the epidermal growth factor receptor. This homology was limited to the region of erbB encoding the tyrosine kinase domain. It was concluded that the neu gene is a distinct novel gene, as it is not coamplified with sequences encoding the EGF receptor in the genome of the A431 tumor line and it maps to human chromosome 17. PMID- 2992091 TI - Isolation and propagation of a human enteric coronavirus. AB - Coronavirus-like particles were found by electron microscopy in stools from infants with necrotizing enterocolitis. Stool samples from these infants as well as control specimens were passaged in cultures of human fetal intestinal organs. Two samples yielded virus-like particles and these have now been passaged 14 times (HEC 14). Gradient-purified HEC 14 strains had typical coronavirus morphology on electron microscopy and contained five major proteins with molecular sizes ranging from 190 to 23 kilodaltons. Infants with necrotizing enterocolitis developed specific antibody to the viral antigens between the acute and convalescent stages of the disease, as shown by examining serum specimens by single radial hemolysis, immunoenzymatic assay, and Western immunoblotting. No cross-reactivity was shown with other coronavirus strains tested, or with the newly isolated viruses of the Breda-Berne group, responsible for calf or horse diarrhea. PMID- 2992093 TI - Gonadotropin-releasing hormone binding sites in human breast carcinoma. AB - Gonadotropin-releasing hormone analogs can cause regression of hormone-dependent breast carcinomas. These effects are thought to be mediated through the inhibition of gonadotropic and steroid hormones. These analogs may also act directly on the tumor because they are effective in treating breast cancer in some postmenopausal women. The presence of specific binding sites for gonadotropin-releasing hormone was demonstrated in human breast carcinomas by means of a novel approach of ligand immunoblotting. The results indicate a possible mechanism by which the peptide has direct effects on this tissue. These binding proteins were not detectable in non-neoplastic breast tissue. PMID- 2992092 TI - Recombinant vaccinia virus: immunization against multiple pathogens. AB - The coding sequences for the hepatitis B virus surface antigen, the herpes simplex virus glycoprotein D, and the influenza virus hemagglutinin were inserted into a single vaccinia virus genome. Rabbits inoculated intravenously or intradermally with this polyvalent vaccinia virus recombinant produced antibodies reactive to all three authentic foreign antigens. In addition, the feasibility of multiple rounds of vaccination with recombinant vaccinia virus was demonstrated. PMID- 2992095 TI - Hepatic receptors and their ligands: problems of intracellular sorting and vectorial movement. PMID- 2992094 TI - Cellular calcium homeostasis and the calcium messenger system. PMID- 2992096 TI - Regulatory factors in pathologic processes of the liver modulators and interacting metabolic networks. PMID- 2992097 TI - Benign fibrous histiocytoma of the left 8th rib. Case report 328. PMID- 2992098 TI - Hepatic resection in cirrhotic patients. AB - Cirrhotic patients are at increased risk for the development of hepatocellular carcinoma, and surgical resection of the involved liver represents the only effective mode of therapy. The presence of cirrhosis significantly complicates hepatic resection. We describe the clinical course of a patient with ruptured hepatoma localized to the lateral segment of the left lobe. This case illustrates many of the problems encountered during hepatic resections in cirrhotic patients, which should be limited to segmental or subsegmental resections. Preoperative attention to bleeding and nutrition, meticulous intraoperative hemostasis, and postoperative management of any pulmonary complications, ascites, and renal or hepatic failure are mandatory for successful management. PMID- 2992099 TI - Japanese encephalitis in Thai children, Bangkok, Thailand. AB - During a period of 5 years from May 1979 to April 1984, 121 encephalitis patients were admitted to Bangkok Children's Hospital. 79 were proved to be JE. 73.42% of JE patients came from outside Bangkok while the rest came from suburban areas of Bangkok. No distinction could be made between JE and encephalitis caused by other agents by clinical data and cerebrospinal fluid examination. Definite diagnosis of JE could be done by demonstration of specific IgM antibody against JEV in cerebrospinal fluid. Case fatality rate of JE was 25%. PMID- 2992100 TI - Elevated spontaneous mutation rate in SV40-transformed Werner syndrome fibroblast cell lines. AB - Spontaneous mutation rates of the cells from patients with Werner syndrome were examined, and we found that the spontaneous mutation rates at the hypoxanthine guanine phosphoribosyltransferase locus in SV40-transformed Werner syndrome cell lines were markedly elevated, compared to those in SV40-transformed normal control cell lines. Our results suggest that Werner syndrome is a mutation mutant. PMID- 2992101 TI - Identification and localization of DNA alteration in Chinese hamster ovary cell mutants (Urd-) defective in the first three enzymes of de novo pyrimidine synthesis. AB - In animals, the first three enzymatic steps of de novo pyrimidine synthesis, carbamyl phosphate synthetase, aspartate transcarbamylase, and dihydroorotase, comprise the multifunctional protein known as the CAD protein. Mutants of Chinese hamster ovary cells (CHO-K1, pro-) deficient in CAD protein activities require uridine for growth and are designated Urd-A mutants. To examine further the nature of the genetic alterations in Urd-A mutants and revertants, we have performed a detailed Southern blot hybridization analysis of DNA from wild-type, Urd-A, and revertant cells using as hybridization probes cDNAs complementary to CAD mRNA isolated from Syrian hamster. This has allowed us to identify an apparent alteration in the CAD gene in DNA from Urd-A cells. This alteration is in a region of the gene which appears to correspond to the region of the protein which is hypersensitive to proteases and which seems to be altered in the mutants. Only one of the two CAD alleles present appears to be altered in this way. Study of certain revertants of Urd-A strongly suggests that in some cases reversion has occurred by amplification of the mutant CAD allele. PMID- 2992102 TI - The acquired immune deficiency syndrome and related complex. A report of 2 confirmed cases in Cape Town with comments on human T-cell lymphotropic virus type III infections. AB - We report a case of acquired immune deficiency syndrome (AIDS) and one of AIDS related complex presenting in Cape Town. The first patient was probably infected in the USA. In turn he infected the second patient by regular homosexual contact. Human T-cell lymphotropic virus type III (HTLV-III) was cultured, we believe for the first time in Africa, from the peripheral blood lymphocytes and a lymph node of our patient with AIDS. HTLV-III infection and high-risk groups in South Africa are discussed in comparison with those in the USA. It is suggested that HTLV-III infection and AIDS will increasingly affect women. Prevention of the spread of HTLV-III infection and AIDS is discussed in relation to close medical surveillance and the protection of blood and blood products from contamination. Counselling of patients with AIDS and persons infected with HTLV-III, general health education, and the protection of health care staff are important in preventing spread but beyond the scope of this article. PMID- 2992103 TI - Isolation of a lymphadenopathy-associated virus from a patient with the acquired immune deficiency syndrome. AB - A virus similar to the lymphadenopathy-associated virus or human T-lymphotropic virus type III, which has been described in association with the acquired immune deficiency syndrome (AIDS) by several laboratories elsewhere in the world, was isolated from a Cape Town patient with lymphadenopathy and acquired immune deficiency. This virus has the characteristic morphogenesis and ultrastructure and its genome encodes the virus-specific p24 protein. It is T-lymphotropic and produces the characteristic cytopathic effect. It can be serially propagated in a human lymphocyte line of the T4+ phenotype. This isolate is being used in diagnostic immunofluorescence assays for virus-specific antibodies. PMID- 2992104 TI - Diet and bowel diseases--past history and future prospects. AB - Changes in diet from ancient times until the present are described. Previously relatively low in energy and animal products yet high in fibre-containing foods, diets are now high in energy and animal products (particularly fat), yet contain less fibre. The changing incidences of bowel disorders and diseases are described, with assessments of the role of diet. Clearly, diet is implicated as regards predisposition to constipation, appendicitis, colorectal cancer and diverticular disease; however, a meaningful dietary role in irritable bowel syndrome, ulcerative colitis and Crohn's disease is doubtful. In South Africa the rarity of bowel diseases in rural blacks compared with whites affords valuable aetiological information about some bowel diseases. The low occurrence thereof (except inflammatory bowel disease) in Indian and coloured populations is not readily explicable. While dietary changes in whites are being widely urged in order to combat degenerative diseases, the magnitude of changes made is unlikely to reduce the occurrence of bowel diseases. The progressive westernization of the diets and lifestyles of less-privileged populations is likely to be associated with increases in the incidences of these diseases. PMID- 2992105 TI - Human cytomegalovirus infection. AB - Human cytomegalovirus (CMV) infection is a very common lifelong infection in South Africa, with intermittent periods of silent reactivation. Infection is spread by intimate contact with infected body fluids (urine, saliva, milk, semen and blood) and most commonly takes place silently in childhood. Clinical disease is rare but may be associated with one or more of the following: (i) a large dose of virus; (ii) a primary infection; and (iii) infection at a time of poor or impaired cellular immunity (developing fetus, after organ transplantation). CMV is an important but rare cause of congenital disease involving especially hearing and neurological deficits. It is often found after organ transplantation but its precise role is not clear. Interpretation of cytomegalovirus tests in connection with an undiagnosed illness is rendered very difficult by the ubiquitous, silently reactivating nature of the virus in both health and disease. An aetiological role is easily assumed but not easily proved. PMID- 2992106 TI - Itraconazole treatment of murine aspergillosis. AB - ICR mice were challenged with conidia of Aspergillus fumigatus given either intranasally or intravenously, and treated beginning 1 day later with itraconazole, amphotericin B, or the vehicles for these two agents. Mice challenged intravenously and treated with either antifungal drug had prolonged survival over controls, and had lower tissue counts in the kidneys than the controls. However, mice challenged intranasally had neither prolonged survival nor lower lung tissue counts than the controls. PMID- 2992107 TI - A noninvasive method for monitoring the effects of increased intracranial pressure with near infrared spectrophotometry. AB - Two experiments were conducted to assess the feasibility of near infrared spectrophotometry (niroscopy) to monitor directly the effects of increased intracranial pressure on brain metabolism. ICP was increased in cats by subarachnoid infusion of a "mock" CSF solution. Cytochrome a,a3 redox state, oxyhemoglobin, deoxyhemoglobin and cerebral blood flow were noninvasively and continuously monitored by niroscopy. The results of both experiments indicated that changes in ICP correlated with a reduction in cytochrome a,a3 redox state (p less than 0.01), a decrease in the quantity of oxyhemoglobin and cerebral flow (p less than 0.01) and an increase in deoxyhemoglobin. The study results suggest that niroscopy has the potential for providing noninvasive and continuous data for assessing brain metabolic activity. The correlations obtained with simultaneous measurements of intracranial pressure make this an attractive method for application to those at risk for increased intracranial pressure. PMID- 2992108 TI - Plastic unification of a double uterus and the outcome of pregnancy. AB - Habitual abortion and premature delivery are occasionally associated with a bicornuate or septate type of uterus when the abnormality is severe. A considerable improvement of reproductive performance was seen in patients who underwent plastic operation of the uterus. The procedures introduced by Strassman, Jones and Tompkins were used. Although it is still difficult to compare the postoperative reproductive history of these three surgical procedures, the procedure used by Tompkins may be technically more simple and provides for an easier entrance into the endometrial cavities in order to incise the common lower uterine segment. Primary sterility still remains a controversial indication for metroplasty and the postoperative reproductive history for this is poor. PMID- 2992109 TI - Breast cancer in a patient with prolactinoma. AB - Although the role of prolactin in the initiation and promotion of mammary carcinoma has been proved in animal experiments, a similar effect of hormone in the development of human mammary tumor has not been clearly demonstrated. If hyperprolactinemia is a significant factor in the initiation and promotion of human breast carcinoma, the incidence of mammary carcinoma in patients with hyperprolactinemia, with or without prolactinomas, might be higher than in control populations, but this relationship has never been documented. We present this case to stimulate awareness of the possibility so the condition will be investigated further. PMID- 2992110 TI - [Effective methods in the medical treatment of osteoporosis]. PMID- 2992111 TI - Treatment policy of surgery in small cell carcinoma of the lung: retrospective analysis of a series of 874 consecutive patients. AB - A therapeutic policy of surgery in the treatment of small cell carcinoma of the lung was evaluated in a retrospective series of 874 consecutive patients. All patients were included in one of six clinical trials of intensive combination chemotherapy and radiotherapy, carried out during 1973-81. General criteria of resectability, including normal appearances at mediastinoscopy, were fulfilled in 33 electively operated patients and in 46 non-operated patients. An elective operation was defined as a thoracotomy performed with the intention of pulmonary resection in a patient known to have small cell carcinoma. Pneumonectomy or lobectomy was performed in 73% of the electively operated patients. Residual macroscopic or microscopic tumour tissue was left after half of these resections. Both operated patients and non-operated patients with operable tumours survived significantly longer than 696 patients deemed to have been inoperable. There was no significant difference, however, in the duration of survival between the two subsets of operable patients. According to this analysis, a treatment policy of surgery for resectable small cell carcinoma does not lead to better overall results than treatment with chemotherapy and radiotherapy alone. PMID- 2992112 TI - HTLV-I antibody status in hemophilia patients treated with factor concentrates prepared from U.S. plasma sources and in hemophilia patients with AIDS. AB - Serum samples from 85 Austrian hemophilia patients treated with lyophilized factor concentrates prepared from U.S. plasma sources, 24 hemophilia patients from Georgia on a home therapy program with factor concentrates, and 10 U.S. hemophilia patients with acquired immunodeficiency syndrome (AIDS) were analyzed by two different methods for the presence of antibodies to the major internal antigen of human T-cell leukemia virus I (HTLV-I) p24. All but one, a Georgia sample, were negative. The absence of antibody to HTLV-I p24 in the serum of European hemophilia patients, of U.S. hemophilia patients with no symptoms of AIDS, and of U.S. hemophilia patients with AIDS is interpreted as an indication of the lack of ready transmissibility of HTLV-I in lyophilized factor concentrates. PMID- 2992113 TI - Agglutination of platelet-type von Willebrand's disease platelets by bovine von Willebrand factor. AB - It has been shown that platelets from patients with platelet-type von Willebrand's disease (vWD) agglutinate upon the addition of human von Willebrand factor (vWF) in the absence of ristocetin or botrocetin, suggesting that platelet membrane receptors for human vWF is abnormal. The present work reports the platelet agglutinability on stimulation with bovine vWF in platelet-type vWD. Platelets in patient platelet-rich plasma or washed platelet suspensions and patient platelets treated with formalin agglutinated in the presence of markedly lower concentrations of bovine vWF than those required for normal platelets. This finding provides additional evidence that platelet-type vWD platelets have abnormal expression of binding sites for vWF on their surface, and supports that platelet receptors for bovine vWF are identical or very close to those for human vWF. PMID- 2992114 TI - Incorporation of some eicosanoic acids into endothelial cells--effect on platelet inhibitory activity and prostacyclin production. AB - Primary cultures of endothelial cells from human umbilical veins were grown until confluency. Then, dihomogammalinolenic acid (DHLA or 20:3n-6) and eicosapentaenoic acid (EPA or 20:5n-3), precursors of monoenoic and trienoic prostanoids, respectively, as well as 5,8,11-eicosatrienoic acid (20:3n-9), and isomer of DHLA, were incorporated into endothelial lipids. DHLA-rich endothelial cells had a decreased capacity of prostacyclin production. By contrast EPA- or 20:3n-9-rich endothelial cells were comparable to controls in this respect. DHLA and EPA were efficiently acylated into cell phospholipids and triglycerides at the opposite of 20:3n-9. It is suggested that both DHLA and EPA could alter the liberation of endogenous arachidonic acid for prostacyclin synthesis but this might be counterbalanced in EPA-rich endothelial cells by PGI3 production. We conclude that DHLA enrichment of endothelial cell lipids may impair the possible beneficial effect of the acid upon platelet functions whereas that of EPA would not be modified. PMID- 2992115 TI - Collagen induced platelet aggregation: a comparison between several commercially available collagens. PMID- 2992116 TI - An evaluation of suloctidil in the prevention of deep vein thrombosis in neurosurgical patients. AB - Suloctidil (200 mg t.i.d.) was compared with placebo in a randomized, double blind trial to assess its value in preventing deep venous thrombosis (DVT) in high-risk neurosurgical patients, comprising 136 patients with brain or spinal tumour, head or spinal injury, or subarachnoid or intracranial hemorrhage. 125I fibrinogen leg scanning and impedance plethysmography were performed for up to 14 days to detect DVT. The two groups were also evenly balanced for DVT risk factors. Seventeen of 68 patients (25%) (95% confidence interval, 15-35%) treated with suloctidil and 12 of 68 patients (21%) (95% confidence interval, 11-32%) treated with placebo developed deep venous thrombosis. This observed difference in outcomes is not statistically significant (X2 = 1.096; p = 0.30). The estimated 95% confidence interval for the true difference in the incidence of DVT between suloctidil-treated and placebo-treated patients ranges from an 11% benefit in favour of suloctidil to an 18% benefit in favour of placebo. Major deep vein thrombosis occurred in two patients on suloctidil and three patients in the placebo group; there were no fatal pulmonary emboli during the 14-day study period, during which time four patients in each group died of non-thromboembolic complications. There was no observed difference in hemorrhagic complications. Long-term outcomes at three-months follow-up were similar between the two treatment groups. It is concluded that there is no real evidence that suloctidil (200 mg t.i.d.) is an effective regimen for the prevention of DVT in high-risk neurosurgical patients. PMID- 2992119 TI - Secretion of glucagon in liver cell carcinoma. AB - In order to elucidate the response of plasma glucagon in liver cell carcinoma, a clinical study was performed in 12 patients with liver cell carcinoma in addition to 8 patients with liver cirrhosis and 8 normal subjects. Arginine infusion elicited increases in plasma insulin and glucagon in 6 patients with liver cell carcinoma as well as 8 patients with liver cirrhosis compared with the controls. However, the responses of plasma insulin and glucagon in liver cell carcinoma did not exceed those in liver cirrhosis. No glucagon secreting cell was proved in the hepatic cancer tissues from two other patients. Furthermore, no measurable glucagon was demonstrated in the tumor tissues extracted from four other patients with liver cell carcinoma. The extract of the tumors, infused into the pancreatic artery of anesthetized dogs, did not elicit any discernible changes in glucagon and insulin in the pancreatic vein. The present study demonstrates an elevated response of plasma glucagon in liver cell carcinoma. Since the morphological and biochemical studies failed to demonstrate the glucagon secreting cell or glucagon stimulating material in the tumor tissues, the elevated plasma glucagon response might be interpreted by the increased A-cell function of the pancreas and the decreased degradation of the hormones in the liver. PMID- 2992118 TI - The effects of PGE2 on vasopressin--and guanine nucleotide-mediated adenylate cyclase activity in toad bladder membrane. AB - PGE2 inhibited 10 m U/ml vasopressin-induced osmotic water flow of the toad bladder at 2 X 10(-8) M. PGE2 suppressed vasopressin-mediated cyclic AMP accumulation in epithelial cells and also vasopressin-mediated adenylate cyclase activity in a crude homogenate of the cells. However, PGE2 had no effect on cyclic AMP dependent and independent protein phosphorylation. These findings indicate that PGE2 inhibits vasopressin-induced water flow mainly through suppression of adenylate cyclase activity, and that the role of PGE2 at that point in the reaction leading to increased water permeability following cyclic AMP production may be slight. Under conditions in which the hormone and substrate are depleted, PGE2 and guanine nucleotides, such as GTP and Gpp(NH)p, additively bring about an increase in adenylate cyclase activity. PMID- 2992117 TI - [Pathology of the digestive tract in kangaroos. A review based on our own study results]. AB - The present paper describes spontaneous pathological findings including etiological aspects in digestive tracts of kangaroos, which have been detected in 166 necropsies during the last 20 years. Protozoan - infections, herpes virus infections, gastroenteritis of unknown etiology and especially the occurrence of the so called "lumpy-jaw" turned out to be of special importance. These findings are discussed with reference to the literature, completed as well by short literature reviews, sub-divided into the different organs and the variable etiology of the diseases, as by a brief description of digestive tract physiology and anatomy in marsupials. PMID- 2992120 TI - Characterization of the effects of erythromycin estolate and erythromycin base on the excretory function of the isolated rat liver. AB - To investigate the mechanisms of erythromycin cholestasis, the effects of erythromycin estolate (EE) on the excretory function of the isolated perfused rat liver and on liver plasma membrane (LM) preparations were studied and compared to those of erythromycin base (EB) and lauryl sulfate (LS), added alone or in combination. EE (at 125 to 200 microM) caused dose-dependent reductions of bile and perfusate flows, bile acid (BA) excretion, and biliary BA concentration. The alterations of the excretory function were only in part due to the decreased perfusate flow. In contrast, both 200 and 300 microM concentrations of EB elicited similar choleretic responses, which were presumably related to the osmotic activity of the drug excreted in the bile. LS did not affect hepatic excretory functions. However, the simultaneous addition of EB and LS resulted in a rate of bile flow lower than that observed with EB alone. EE, but not EB, increased canalicular permeability to [14C]sucrose as measured by bile to plasma (B:P) ratio. Neither drugs altered [14C]erythritol B:P ratio. In LM preparations both Na+,K+- and Mg2+-ATPase activities were inhibited in a dose-dependent manner by EE, but not by EB. The data suggest that EE could affect bile flow by inhibiting cotransport of Na+ and BA and by altering LM permeability and support the view that the effect of erythromycins on the liver may be related to their surface activity. PMID- 2992121 TI - Changes in adrenal function as a possible mechanism for elevation of serum glucose by a single large dose of fluoride. AB - Serum glucose was elevated immediately after ip administration of a single large dose of fluoride (NaF 35 mg/kg) to rats. Moreover, elevation of serum glucose following ip administration of 35 mg/kg of fluoride to rats was suppressed by adrenalectomy, dibenamine, or propranolol, but not by thyroid-parathyroidectomy. The elevation of serum glucose was associated with enhancement of glucose-6 phosphatase activities in liver and kidney in fluoride-treated rats. PMID- 2992122 TI - Fractionation of red diamond rattlesnake (Crotalus ruber ruber) venom: protease, phosphodiesterase, L-amino acid oxidase activities and effects of metal ions and inhibitors on protease activity. AB - Crotalus ruber ruber venom contains several different proteases, and the proteolytic activity of the crude venom is 6-15 times greater in adult than in juvenile venom. Venom samples were assayed for proteolytic, phosphodiesterase, L amino acid oxidase and elastinase-like activities and were subjected to gel filtration on BioGel P-100. Two major size classes of proteases were resolved (mol. wt 67,000 and 20,500). EDTA, N-ethylmaleimide (N-EM) and 1,10 phenanthroline inhibited proteolytic activity of crude venom, and EDTA, Zn2+ and Cu2+ inhibited proteolytic activity of the fractionated venom. PMID- 2992123 TI - Neurological analysis of respiratory, cardiovascular and neuromuscular effects of brevetoxin in cats. AB - Effects of brevetoxin were evaluated in cats anesthetized with pentobarbital under conditions of controlled end-expiratory pCO2 and constant body temperature. Recordings were made of arterial blood pressure, heart rate, respiratory pattern, diaphragm EMG, evoked tibialis muscle twitch and evoked contraction of the nictitating membrane. Electrical stimulation was employed for periodic excitation of the medullary respiratory center, the phrenic nerve, the peroneal nerve and the cervical sympathetic nerve. Brevetoxin was prepared at a concentration of 1.0 mg/ml in an aqueous medium of 2.5% ethanol plus 2.5% Emulphor 620 (General Aniline and Film Corp., New York). Small i.v. bolus injections of the toxin (40 micrograms/kg) evoked, without tachyphylaxis, the Bezold-Jarisch reflex triad of bradycardia, hypotension and bradypnea. This effect was essentially abolished by vagotomy. Continued injections then resulted in pressor reactions and tachycardia, along with the development of respiratory dysrhythmia. Large doses of brevetoxin (160 micrograms/kg i.v.) caused somatomotor seizures accompanied by severe hypertension, that occurred even after decerebration and cervical spinal cord transection. Cranial intra-arterial and intra-cerebroventricular injections of brevetoxin produced hypertension and respiratory depression more effectively than did i.v. injections. Systemic cumulation of the toxin, with the respiration supported artificially, caused death from cardiovascular collapse, without significant blockade of neuromuscular and ganglionic transmission. It is concluded that brevetoxin exerts its major toxic effects on the circulation and respiration through reflex and central actions, largely sparing peripheral motor mechanisms. PMID- 2992124 TI - Postsynaptic blocking of glutamatergic and cholinergic synapses as a common property of Araneidae spider venoms. AB - Venom effects of eight Araneidae spider species were studied using locust and frog neuromuscular junctions. The spider venoms irreversibly blocked miniature excitatory postsynaptic potentials and excitatory postsynaptic potentials of locust neuromuscular junction. The frog miniature end-plate potentials and end plate potentials were also blocked, but they recovered upon washing of the preparation with physiological solution. PMID- 2992125 TI - In vitro and in vivo effects of vanadate on K+-dependent phosphatase activities from subcellular fractions of brain, kidney and liver. AB - Inhibition of K+-dependent phosphatases by sodium orthovanadate was studied in the subcellular fractions of brain, kidney and liver from male Sprague-Dawley rats. The concentration required to produce an in vitro phosphatase inhibition of 50% ranged from a low of 0.2 microM in the kidney 900 X g fraction to a peak of 400 microM in the brain soluble fraction. The concentrations required to produce 50% inhibition occurred in the micromolar range for most fractions studied. Injection of sodium vanadate (i.p., 8 mg/kg) produced phosphatase inhibition in several of the fractions, but not as pronounced as the in vitro treatment. The results indicate that vanadate is a potent inhibitor of phosphatase activity in vitro, and may have similar potential in vivo. PMID- 2992126 TI - [Prospects for using the electron paramagnetic resonance method in forensic medicine]. PMID- 2992127 TI - Ocular disease in immunosuppressed patients. AB - New therapies and diseases causing immunosuppression have provoked new and devastating ocular diseases. The possible reasons for the vulnerability of the retina to opportunistic infections are discussed. The clinical patterns of disease caused by common opportunistic agents are described, and current treatment available is reviewed. PMID- 2992128 TI - Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells. AB - Seropositivity to human T-cell lymphotrophic virus-III (HTLV-III) can have profound implications for the individual in whom it is detected. Simple and reliable tests are needed to confirm positivity by screening assays. In this study, detection of human antibodies by indirect immunofluorescence (IFA) on acetone-fixed HTLV-III infected H9 cells was evaluated in blood donors, patients with infectious or auto-immune diseases, and men with or at high risk for developing acquired immune deficiency syndrome (AIDS). Specific and nonspecific patterns of immunofluorescent reactivity were distinguished easily. None of 98 serums from blood donors was positive, while two of 33 serums from patients attending an infectious disease clinic, both homosexual men, were positive. Ninety-six percent of 24 serums from men with AIDS, 87 percent of 70 serums from men with lymphadenopathy, and 50 percent of 135 serums from healthy gay men were positive. These results paralleled those obtained by Western blotting and membrane immunofluorescence. In contrast, 11 and 4 percent, respectively, of these serums were judged as borderline or not interpretable by an enzyme-linked immunosorbent assay (ELISA). Of these serums, those that were positive by IFA were positive by Western blots, and 16 of the 17 IFA-negative serums were negative by Western blots. These studies indicate that IFA is a sensitive and specific assay for HTLV-III antibodies in human serums. PMID- 2992129 TI - Anti-HTLV-III testing of blood donors: reproducibility and confirmability of commercial test kits. AB - Since 2% of the cases of Acquired Immune Deficiency Syndrome (AIDS) have been attributed to transfusions of blood and blood products, licensed tests to detect antibody to the human T-lymphotropic virus type III (anti-HTLV-III) have been put into practice to reduce the risk of transfusion associated AIDS. Two commercial ELISA kits (Abbott and ENI) were used to test for anti-HTLV-III in 100 coded samples from individuals with AIDS, at high risk for AIDS, or with low risk for AIDS and in 1280 unlinked blood donor serums. From the 100 coded samples, both Abbott and ENI tests identified 51 of 52 coded samples with anti-HTLV-III which were confirmable with Western blot analysis. Initial testing of the donor serums by Abbott's test revealed 20 reactives, of which 5 were repeatably reactive; initial testing by ENI's test revealed 25 reactives, of which 14 were repeatably reactive. However, only 3 donor serums were repeatably reactive by both test kits, out of 17 repeatable reactive by either, and no ELISA positive samples were confirmed by Western blot or IFA. Before a blood donor is notified of "anti-HTLV III reactivity", tests demonstrating this should be both reproducible and confirmable by at least one additional test. PMID- 2992130 TI - The role of pretransplant immunity in protection from cytomegalovirus disease following renal transplantation. AB - To determine the extent to which pretransplant immunity resulting from natural infection protects against cytomegalovirus (CMV) disease, we analyzed CMV serology on 153 kidney donor and recipient pairs and followed transplant patients to determine incidence and severity of CMV disease. The overall incidence of CMV disease was 22%. Significant differences occurred in CMV disease incidence and severity, depending on the immune status of the kidney donor and recipient. Among recipients of kidneys from seropositive donors, immunity offered significant protection from CMV disease, reducing its incidence from 61% in nonimmune to 24% in immune patients (P less than 0.01). Pretransplant immune patients also had fever CMV-related complications. Among recipients of kidneys from seronegative donors, pretransplant immunity conferred a significant risk of CMV disease; immune patients had a 20% incidence of CMV disease compared with 2% in nonimmune patients (P less than 0.02). Disease was generally mild in all patients receiving kidneys from CMV infection had a 3-fold higher incidence of CMV disease than patients with reactivation infection (P less than 0.01). The incidence of CMV disease was similar in immune patients, whether they received a kidney from a seropositive or a seronegative donor. However, an important observation was that disease was significantly more severe in immune patients receiving a kidney from a seropositive donor (P less than 0.05). This indicates that if kidneys from seropositive donors are selected for use only in seropositive recipients, this places the immune patient at a higher risk for severe CMV disease. We conclude that pretransplant immunity offers a significant advantage to patients receiving kidneys from seropositive donors. PMID- 2992131 TI - Effect of captopril and theophylline treatment on cyclosporine-induced nephrotoxicity in rats. PMID- 2992132 TI - Cytomegalovirus infection in renal transplant patients. PMID- 2992133 TI - Hemodynamic effects of hypertonic sodium bicarbonate in canine cardiac allograft recipients during acute rejection. PMID- 2992134 TI - Unusual clinical presentations of hepatocellular carcinoma. PMID- 2992135 TI - The effects of westernisation on sigmoid volvulus. PMID- 2992136 TI - [Genetic transformation of somatic cells. V. Inheritability of the rate of loss of the trait and the stabilization of the transformant phenotype]. AB - Subclones were isolated both on selective and nonselective medium from the Chinese hamster cells transformed by thymidine kinase gene (TK-gene) of Herpes simplex virus (HSV-1) and varying in the rate of the loss of transformant phenotype. The study of the stability of thymidine kinase-positive (TK+) phenotype in cell populations the subclones shows that the nonstability and the rate of the loss of transformant phenotype are the characters that are inherited in the cell generations. Durable cultivation on a HAT-selective medium may lead to a complete or partial (expressed as a reduced rate of the loss of the character) stabilization of TK+-phenotype of the cells of transformant clones. The rate of stabilization of TK+-phenotype may differ depending on the structure of transforming DNA introduced into cells of transformant clones. PMID- 2992137 TI - [Genetic transformation of somatic cells. VI. Transfer of the trait of the rate of loss of transformant phenotype by means of DNA from cells containing the thymidine kinase gene of the herpes simplex virus]. AB - Using dot-hybridization with thymidine kinase gene (tk gene) of Herpes simplex virus type 1 (HSV 1) of DNA preparations obtained from isolated metaphase chromosomes and lysate fractions of metaphase cells, which presumably contain smaller particles compared to metaphase chromosomes, it has been shown that the tk gene of HSV 1 is localized in chromosomes of cells of transformant clones unstable in TK+-phenotype. The DNA isolated from the metaphase chromosomes from cells of transformant clones is 1.5- or 2-fold more efficient in transforming TK Chinese hamster cells than is the total high molecular weight DNA from the same cells. Upon transformation of TK- cells by the high molecular weight DNA from the tk gene of HSV 1-containing clones, varying in the rate of the loss of TK+ phenotypes, the character "rate of the loss of transformant phenotype" is transferred together with the tk gene of HSV 1 in 22% of cases. Cells of rerevertant clones, produced from TK- subclones of transformant clones, display the rate of the loss of transformant phenotype characteristic of cells of parental TK+-clones. A comparison of the results allows a conclusion that DNA sequences, determining the character "rate of the loss of transformant phenotype", are linked tightly with the transforming DNA proper containing the tk gene of HSV 1, but are not localized inside such a DNA. PMID- 2992138 TI - Antibody to bluetongue virus in cattle in Ecuador. PMID- 2992139 TI - Detection of foot-and-mouth disease virus antibody using counterimmunoelectrophoresis and serum neutralisation tests. AB - A comparative investigation was made on the applicability, sensitivity and specificity of counterimmunoelectrophoresis (CIEP) for the rapid detection of antibody to foot-and-mouth disease virus in cattle sera using as reference a standard serum neutralisation test. The CIEP test was sensitive and exhibited a reasonable specificity. PMID- 2992141 TI - [Changes in the activity of enzymes of renin-angiotensin and kinin systems in the rat brain after adrenalectomy and administration of hydrocortisone]. AB - It is shown that the activity of enzymes participating in renin-angiotensin and brain kinin systems' metabolism depends on functional state of hypothalamo pituitary-adrenocortical system. Under experimental hypocorticism the activity of angiotensin-converting enzyme and kininase I in the hypothalamus, hippocamp, corpus striatum and rat pituitary decreases; the renin-like enzyme activity decreases in the corpus striatum but increases in the hypothalamus and hippocamp. After hydrocortisone administration to adrenalectomized rats the angiotensin converting enzyme activity of the hippocamp and pituitary is shown to be normalized as well as renin-like enzyme and kininase I of the hippocamp and corpus striatum. The activity of the studied enzymes in the hypothalamus decreases in this case. PMID- 2992140 TI - [Isolation and properties of 2 forms of cyclic nucleotide phosphodiesterase from the rat brain under normal conditions and after irradiation]. AB - It is established that the functional activity of two phosphodiesterase forms- phosphodiesterase I (Ca2+-calmodulin-sensitive) and phosphodiesterase II (Ca2+ calmodulin-insensitive), isolated from grey matter of the irradiated rat brain varies essentially in comparison with that of the normal rats. In the early period of acute radiation injury both phosphodiesterase I sensitivity to calmodulin and phosphodiesterase II special activity under hydrolysis of 3', 5' GMP decrease but phosphodiesterase I special activity under hydrolysis of 3', 5' GMP increases. The investigation of temperature dependence of phosphodiesterase I and phosphodiesterase II activations revealed changes in character of curves, the temperature optimum under irradiation being unchanged and inflections appearing on the Arrhenius curves. PMID- 2992142 TI - [Na+,K+-ATPase activity and potassium, sodium and calcium levels in the rat myocardium and brain during pain-induced emotional stress]. AB - An emotional-algesic stress in the period of its development and after-effect causes a different-directed influence on the state of the ionic transport in the heart and brain of rats. The Na+, K+-ATPase activity in the left ventricle of the heart decreases with a simultaneous increase in the sodium content and decrease in the calcium and potassium content. The Na+, K+-ATPase activation is observed in the cortex of cerebral hemispheres with a simultaneous increase in the content of mentioned ions. PMID- 2992144 TI - [Calcium tolerance test in normal subjects]. PMID- 2992143 TI - [Localization of insulinomas: the value of determining insulin using transhepatic catheterization of the pancreatic venous network]. PMID- 2992145 TI - Intraoperative consultation for the retroperitoneum and adrenal glands. AB - An incidentally discovered mass lesion is the most frequent indication for intraoperative consultation involving the retroperitoneum or adrenal gland. The goal of the surgeon is to determine the nature and extent of the lesion and, for solid lesions, to obtain a biopsy adequate for histopathologic diagnosis. Benign lesions may be excised locally, whereas therapy for malignancies depends upon the individual circumstances of the patient, the histology of the lesion, and involvement of adjacent organs. Treatment of hematomas requires early vascular control. Adrenal tissue should be preserved whenever possible. PMID- 2992146 TI - Sperm antibodies and infertility in patients with testicular cancer. AB - The level of antisperm antibodies using the enzyme-linked immunosorbent assay (ELISA) in the serum of 48 patients with testicular cancer before and after therapy is reviewed. This is not a linear study of each patient, but some conclusions can be drawn: In a high percentage of testicular cancer patients serum antisperm antibodies can be detected: 73.3 per cent before orchiectomy and 43.7 per cent overall. The percentage of patients with antibodies decreases with adequate therapy. In patients with advanced disease there is a higher percentage of positivity (50%) for serum antisperm antibodies than in patients with low stage disease (30%). The higher percentage of antibodies-positive patients among those with infertility patterns could be an important argument that supports the hypothesis that autoimmune pathology can play a role in oligo/azoospermia in testicular cancer patients. PMID- 2992147 TI - Small-cell carcinoma of prostate. Transient complete remission with chemotherapy. AB - A patient with small-cell carcinoma of the prostate is described. The patient's clinical course and the tumor's pattern of metastases were similar to those seen in small-cell carcinoma of the lung. There was a complete, though brief, remission on chemotherapy with cyclophosphamide, doxorubicin hydrochloride (Adriamycin), and vincristine. PMID- 2992148 TI - [Testing the susceptibility of cultured cells to infection with bovine leukemia virus]. AB - Different cell cultures were studied for their susceptibility to bovine leucosis virus infection. Syncytial assay was used for this study. The FLS/BLV+ cell line served as virus source. Cell lines BHK-21 and ZP-1/58 were found to be susceptible to syncytium formation. Large cells with one to three large nuclei, and loose nuclei reaching the size of syncytium were observed to occur in the BHK 21 and ZP-1/58 cell lines, apart from the syncytial formations. The virus specificity of the syncytia arising in these two cell lines was confirmed by the immunofluorescence assay. In the case of the immunoperoxidase assay, a positive result was obtained only in the BHK-21 cell line. The occurrence of syncytia and large nuclei was observed even in the cases when the BHK-21 cells were infected with the lymphocytes of leucotic cows. PMID- 2992149 TI - [Survival of Aujeszky's disease virus in infected pig slurry]. AB - It has been demonstrated that after experimental infection of pig slurry from the space under the slatted floor (infection dose of 10(6)PFU per ml), the Aujeszky's disease virus (ADV) survived for 72 hours at the temperature of 15 degrees C and at pH 6.5, but was inactivated after 96 hours. When technologically treated pig slurry from the storage tanks was saturated with water and infected with ADV at the dose of 10(5)PFU per ml, the virus survived for 23 days when kept at 15 degrees C and 4 degrees C and at pH 6.8, but was inactivated under the same conditions after 30 days. When the infective ADV dose in the technologically treated pig slurry in the storage tanks was reduced to 10(4)PFU per ml, the virus survived 16 days at +4 degrees C and pH 7.0 and 8.0 but was inactivated within 23 days after infection. PMID- 2992150 TI - An ultrastructural morphometric analysis on ultrathin epon and ultrathin cryosections of normal human gastric tissue and human gastric cancer. AB - An ultrastructural morphometric study on ultrathin epon, and cryosections of normal and neoplastic, adult, human, gastric tissue is presented. The data show a characteristic numerical pattern for the different neoplastic types of gastric tissues. PMID- 2992151 TI - Mutations in the herpes simplex virus major DNA-binding protein gene leading to altered sensitivity to DNA polymerase inhibitors. AB - Five herpes simplex virus mutants containing temperature-sensitive mutations in the gene for the major DNA-binding protein were assayed for their sensitivities to the DNA polymerase inhibitors aphidicolin and phosphonoacetic acid (PAA). Four of the mutants (tsA1, tsA15, tsA24, and tsA42) exhibited altered sensitivity to one or both of the inhibitors relative to the wild-type parent. In tsA1, a mutation or mutations conferring aphidicolin and PAA hypersensitivity were mapped by corescue with the temperature-sensitivity marker of tsA1 to a region of the DNA-binding protein locus, between map coordinates 0.385 and 0.398. The mutation conferring PAA hypersensitivity in tsA24 similarly corescued with the tsA24 temperature-sensitivity marker, mapping to the DNA-binding protein locus between coordinates 0.398 and 0.413. Thus, mutations outside the DNA polymerase locus and within the DNA-binding protein locus can confer altered sensitivity to certain DNA polymerase inhibitors. Assays of the aphidicolin and PAA sensitivities of ts+ recombinants derived by marker rescue of the DNA-binding protein mutants revealed the presence of additional mutations, separable from the ts mutations, in each of three mutants examined. One such mutation, which contributed to the aphidicolin hypersensitivity phenotype of tsA1, mapped between coordinates 0.422 and 0.448, and resides, most probably, within the DNA polymerase locus. These additional mutations possibly confer compensating modifications to the DNA polymerase such that functional interaction with altered DNA-binding protein is restored. These findings provide strong evidence that the major DNA-binding protein and the DNA polymerase of herpes simplex virus interact in infected cells. PMID- 2992152 TI - Sequence analysis of the viral core protein and the membrane-associated proteins V1 and NV2 of the flavivirus West Nile virus and of the genome sequence for these proteins. AB - Cell-associated flaviviruses contain the two membrane proteins V3 and NV2 besides the viral core protein V2 whereas extracellular viruses do contain V2 protein and the two membrane proteins V3 and V1. Since the V1 protein could not be detected in infected cells it has been suggested that V1 is generated from NV2 by proteolytic cleavage during the release of virus from cells (D. Shapiro, W. E. Brandt, and P. K. Russell (1972), Virology 50, 906-911). We have isolated the viral structural proteins V1, V2, and NV2 from the flavivirus West Nile virus and determined their amino-terminal amino acid sequences and amino acid sequences of peptides derived from these proteins. We have also transcribed parts of the viral genome into cDNA and cloned and sequenced this cDNA. The analyses of the protein structure of V1, V2, and NV2 together with the determination of the amino terminal sequence of V3 (data not shown) have allowed us to identify the nucleotide region coding for the structural proteins V2, NV2, and V1. The primary structure of this nucleotide sequence is presented in this report. The data show that the amino terminus of the viral core protein V2 is followed by the amino termini of the proteins NV2, V1, and V3, respectively. These data for the first time identify the exact order of all structural proteins of a flavivirus identified so far. Our data strongly support the above-mentioned hypothesis that V1 is derived from NV2 by proteolytic cleavage and furthermore indicate that V1 represents the nonglycosylated carboxy-terminal part of NV2 which contains those sequences which anchor NV2 in the viral membrane. A working hypothesis is presented in which two species of cellular enzymes, signalase(s) removing signal sequences and enzymes involved in cleaving polyproteins after a pair of basic amino acids, do generate the proteins V2, NV2, and V1 from the growing peptide chain synthesized during translation of the 42 S genome RNA which functions as mRNA for these proteins. PMID- 2992153 TI - Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix. AB - A total of eight human epithelial cell lines derived from the carcinoma of the cervix were examined for the presence of human papillomaviruses (HPVs) types 16 and 18 DNA sequences. Six out of eight cell lines contain sequences hybridizing to the DNA of these viruses. Two of the cell lines contain sequences hybridizing specifically to HPV 16. One of these two cell lines contains all of the HPV 16 sequences and the other cell line is missing fragments containing early regions E2 and E4 and some of the late regions. Four of the cell lines contain sequences hybridizing specifically to HPV 18. All these cell lines are missing fragments containing early regions E2, E4, and E5. Interestingly, all the cell lines contain sequences corresponding to early regions E1, E6, and E7. PMID- 2992154 TI - Sites of recombination between the transforming gene of avian myeloblastosis virus and its helper virus. AB - The sites of recombination between the transforming gene of avian myeloblastosis virus (AMV) and its natural helper myeloblastosis-associated virus (MAV) have been determined. In AMV, the cellular sequence substituting for the viral envelope (env) gene gives rise to a different carboxyl terminus of the DNA polymerase. The 5'-recombination site coincides with the RNA splice acceptor site for the production of env mRNA in MAV-infected cells. The 3'-recombination site reveals that the last 11 amino acids including the termination codon are shared by the env protein and AMV transforming protein. The RNA splice acceptor site for the generation of subgenomic v-myb mRNA is located 84 nucleotides downstream from the 5'-recombination site. The AMV transforming protein consists of helper virus related sequences at both of its amino and carboxyl termini, and all but 84 nucleotides of the cell-derived v-myb sequence. The comparison of MAV gp85 amino acid sequence with those of subgroups B, C, and E indicates that the MAV present in clone lambda 10A2-1 belongs to subgroup B. The high degree of homology among different avian retroviruses of the same subgroup indicates that the amino acid sequence of gp85 is important in determining the conformation of the envelope glycoprotein. PMID- 2992155 TI - In vitro growth of two related leporipoxviruses in lymphoid cells. AB - We examined the in vitro growth patterns of two leporipoxviruses, malignant rabbit fibroma virus (MV) and Shope fibroma virus (SFV), in lymphoid cells. MV replicates well in normal spleen cells in vitro. At low m.o.i. (0.001), dramatic virus growth occurs in unstimulated cell cultures. This growth is enhanced by addition of the T lymphocyte mitogen, concanavilin A, or the B lymphocyte mitogen, Escherichia coli lipopolysaccharide. Shope fibroma virus does not grow in lymphocytes in culture, with or without mitogen stimulation. MV itself profoundly inhibits lymphocyte mitogenesis, while SFV does not. MV and SFV added to normal lymphocytes do not appear to alter their viability in culture. Thus, MV appears to be novel in its ability to replicate to high titer in resting lymphocytes. This growth pattern may be useful in understanding MV-induced immunologic dysfunction. PMID- 2992156 TI - Maturation of murine leukemia virus env proteins in the absence of other viral proteins. AB - A mutant of Akv which produces env but no detectable gag or pol products (A. Rein, D. R. Lowy, B. I. Gerwin, S. K. Ruscetti, and R. H. Bassin, J. Virol. 41, 626-634 (1982] was examined for maturation of env gene protein products. In comparison with wild-type Akv, the mutant AK 71 synthesizes gPr85env and produces gp70 and Prp15E in normal amounts and with normal kinetics. Cell surface gp70 was found alike in the mutant and wild type. However, cleavage of Pr15E to p15E did not occur in the mutant. This final cleavage step of AK 71 Prp15E could be made to occur by superinfecting cells containing the mutant with baboon endogenous virus. Thus, unlike earlier steps, this final step in maturation of the env gene product appears to require gag or pol gene products. It is proposed that the virus-encoded protease is required for this last step. PMID- 2992157 TI - Molecular cloning of integrated caprine arthritis-encephalitis virus. AB - A full-length DNA clone of the exogenous retrovirus, caprine arthritis encephalitis virus (CAEV), was isolated from high molecular weight DNA of CAEV infected Himalayan tahr ovary cells. Although other restriction maps of CAEV have been published, this is the first time that the proviral DNA has been cloned. The restriction enzyme map of the clone was determined and found to be identical to that of unintegrated linear CAEV DNA except for the presence of cellular flanking sequences. These findings establish that lentiviruses are able to integrate within the infected host cellular genome. The cloned CAEV genome was shown to contain terminal repeats of approximately 450 base pairs in length, and its restriction enzyme map was oriented with respect to the direction of viral RNA transcription. When the cloned CAEV DNA was used as a molecular probe, it failed to detect related proviral sequences in the genomes of a variety of vertebrate species, including the goat, sheep, horse, mouse, and man. When CAEV DNA was hybridized under relaxed conditions to a variety of cloned DNAs, representing different oncoviral genera, homology to mouse mammary tumor virus (MMTV) was observed, while no homology to avian type C or mammalian type A, C, and D retroviruses was detected. This homology was localized to a region in MMTV corresponding to the 3' end of the gag gene and the 5' end of the pol gene. PMID- 2992158 TI - Polyadenylic acid sequences in the genomic RNA of the togavirus of simian hemorrhagic fever. AB - Since serologic studies have failed to relate the togavirus simian hemorrhagic fever (SHF) virus to any currently accepted genus within the Togaviridae family, the presence of polyadenylic acid [poly(A)] in the genomic RNA was analyzed in view of the different content reported for the two major genera of that family: alphaviruses where poly(A) is 40 to 120 nucleotides long and flavivirus where poly(A) is considered to be absent. Oligo(dT)-cellulose chromatography of whole genomic RNA from purified SHF virus revealed that about 36% of the molecules contained segments of poly(A) of sufficient length to bind to oligo(dT) cellulose. However, a reproducible fraction of the RNAs did not bind to oligo(dT) cellulose, indicating little or no poly(A) present. When analyzed by electrophoresis under denaturing conditions, both the binding and nonbinding molecules were similar in size. In addition, no polyuridylic acid [poly(U)] was detected in SHF virus genomic RNA. After digestion of the genomic RNA with pancreatic and T1 ribonucleases, the resultant resistant polynucleotide sedimented by ultracentrifugation between tRNA and 5 S RNA. Base composition analysis of these polynucleotides detected only adenosinic residues. A mean length of 76 +/- 2 nucleotides for these poly(A) sequences of SHF virus RNA was established by electrophoresis under denaturing conditions. Thus, together with previous morphological as well as biochemical findings, the presence of a poly(A) sequence is further evidence that SHF virus has distinctive characteristics which differentiates it from the two major subgroups of togavirus. PMID- 2992159 TI - Transposon-mediated mutagenesis of a baculovirus. AB - Spontaneous mutants of insect nuclear polyhedrosis viruses of Autographa californica (AcMNPV) and Galleria mellonella (GmMNPV) were analyzed. These mutants produce few polyhedra in infected cells and have insertions of host cell DNA. All the insertions mapped to two adjacent sites in the genome. The junctions between two host insertions and the viral DNA were sequenced. One of the insertions contained a perfect 7-bp inverted terminal repeat, and had caused a direct duplication of 4 bp of viral DNA at the point of insertion. Therefore, this insertion sequence has properties of a transposon of the host cell Trichoplusia ni. PMID- 2992160 TI - Latency of horizontal cell response in the carp retina. AB - The response latency of horizontal cells in the carp retina was investigated by means of the stimulus-response crosscorrelation using a randomly modulated light of various wavelengths. R/G type cells produced two response components with some difference in latency; short-latency hyperpolarising response to green light and long-latency depolarising one to red light. The latency difference could not be made smaller than 14 ms by increasing the intensity of test light. These results were reconciled with similar measurements under selective chromatic adaptation. The latency difference revealed here supports the current view of neural circuitry that accounts for spectral response characteristics of horizontal cell subtypes. PMID- 2992161 TI - [The effect of isoprenaline on the levels of cyclic adenosine monophosphate (cAMP) in the lymphocytes of persons in euthyroid, hyperthyroid and hypothyroid states]. PMID- 2992162 TI - [Na+, K+-ATPase activity of the kidneys and 3,5,3'-triiodothyronine levels in rat plasma during adaptation to cold]. AB - No correlation between Na, K-ATPase activity increase in rat kidney and the level of T3 in blood plasma was revealed in course of the rats adaptation to cold. Spironolactone prevented the enzyme activation caused by cold. Actinomycin D, administered to the acclimated rats two days before the experiment, decreased the enzyme activity. Addition of actinomycin D to membrane preparation did not change the enzyme activity. The increase in Na, K-ATPase activity in kidney during adaptation of rats to cold appears to be related to DNA-dependent synthesis of RNA. The enzyme activation was apparently regulated by aldosterone but it was not the T3-dependent process. PMID- 2992163 TI - [Enzymes of purine nucleotide catabolism in lymphocytes in normal states and in chronic lymphoid leukemia]. AB - Activities of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) as well as their ratio in chronic lymphocytic leukemia (CLL) were found to be several times lower as compared with normal cells and to depend upon the duration and severity of leukemic process. Ratio of ADA and PNP activities in CLL was inverted as compared with those of normal cells; 5'-nucleotidase activity varied within all the stages of the disease from zero values to supernormals. There was a correlation between beneficial effects of treatment of the CLL patients and an increase in ADA and PNP activities in their peripheral lymphocytes. PMID- 2992164 TI - [Changes in Na, K-ATPase activity in subcellular fractions of the lumbar region of the spinal cord in botulism and the possibility of their pharmacological correction]. AB - Inhibition of Na, K-ATPase in fractions of unpurified synaptosomes and of heavy microsomes from lumbar part of spinal cord was found in dynamics of botulinic intoxication of the C type in rats. Both competitive and noncompetitive types of inhibition of the enzymatic activity were found in kinetic studies. Inhibition of transport functions across biological membranes caused by botulinic toxin was shown to have a reversible character. The inhibition might be prevented within the early periods of botulinic intoxication by administration of serotonin and guanidine hydrochloride. PMID- 2992165 TI - [Ultrastructural characteristics of the histological forms of breast tumors]. AB - Differences in ultrastructurally-differentiated and undifferentiated cells distribution were established in such non-invasive and invasive histological patterns of breast cancer as medullar carcinoma, mucinous, adenoid-cystic and lobular invasive cancer by means of electron microscopy. It was shown that a certain histological pattern is not necessarily matched by a relevant degree of ultrastructural differentiation of tumor cells. Tumor tissues of a similar histological pattern sometimes revealed different ratios of ultrastructurally differentiated and undifferentiated cells, which may be responsible for the peculiarities in the clinical behavior of the said neoplasms. These data should be taken into account in establishing ultrastructural diagnosis as well as prognostic evaluation of the findings of electron microscopic examination in cases of non-invasive and certain invasive forms of breast tumors. PMID- 2992166 TI - [Electron microscopic and biochemical characteristics of the hormonal activity of small-cell lung cancer]. AB - An electron microscopic study of small-cell carcinoma of the human lung showed that tumors identified by light microscopy as small-cell lung cancer were, in actual fact, histologically heterogenous, ranging from those free from any signs of specific cell differentiation, neoplasms with endocrine differentiation, squamous cell cancer, adenocarcinomas to tumors with mixed cell differentiation. Elevated blood-ACTH concentrations occur in patients with tumors which do not incorporate cells with endocrine differentiation. In such cases, activation of ACTH synthesis may be due to hyperfunction and hyperplasia of apudocytes (elements of a diffuse endocrine system--the APUD system) of the lung or other organs in response to tumor growth. PMID- 2992167 TI - [Clinical characteristics of a malignant fibrous histiocytoma of the soft tissues]. AB - The results of a study of 51 cases of malignant fibrous histiocytoma of soft tissues are discussed. The survival versus clinico-morphological feature correlation was studied. Shorter survival time was found to be associated with pain syndrome and tumor-produced skin symptoms as well as in cases of tumor misdiagnosed as hematoma. Female patients over 50 years showed relatively longer survival time. No correlation was established between survival time and histological pattern of tumor. PMID- 2992168 TI - [Growth factors and their activation of protein kinases in benign and malignant tumors]. PMID- 2992169 TI - [Association of lung cancer with adenoma of the adrenal cortex]. PMID- 2992170 TI - [Comparison of x-ray and radionuclide studies of the joints in patients with Touraine-Solente-Gole syndrome]. AB - The thesis of primariness of the disease and of its osseous tissue manifestations is discussed on the grounds of the results from a complex study on 3 patients. The periosteal ossifying hyperplasia and disturbances in the osseous tissue structure develop with no other organ symptomatics and in the absence of any changes in the available laboratory and immunological indices. The macromorphological X-ray characteristic corresponds to the radionuclide tests of the study with 99MTc-pertechnetate, 99MTc-pyrophosphate and 99MTc-sulfocolloid, informing about abnormal vascularization index, growth of bone-marrow tissue and intensified joint-tissue anabolism. PMID- 2992172 TI - Inherent contrast in magnetic resonance imaging and the potential for contrast enhancement. The 1984 L. Henry Garland lecture. AB - Magnetic resonance (MR) imaging is emerging as a powerful new diagnostic tool valued for its apparent lack of adverse effects. The excellent inherent contrast between biologic tissues and fluids afforded by MR imaging is one of the foremost characteristics of this technique and depends on physicochemical properties such as hydrogen density and T1 and T2 relaxation rates, on magnetic field strength and on operator-chosen factors for acquiring the MR imaging signal. Pharmaceutical contrast-enhancing agents shorten the MR imaging process and improve sensitivity and diagnostic accuracy. PMID- 2992173 TI - [Results of surgical treatment of common peripheral nerve lesions of the upper extremities]. AB - The results of operative treatment and clinical as well as electrodiagnostic findings are discussed in 68 cases. In about one half of our cases a carpal tunnel syndrome was found with excellent outcome of the operation in 93,6%, with fairly good outcome in the rest of the cases. In 14 of the patients studied a sulcus nervi ulnaris syndrome was seen, 9 of them showing excellent improvement after operative neurolysis was performed. Motor function improved in 8 out of 11 traumatic nerve dissections; Restitutio ad integrum was seen in incomplete lesions of the brachial plexus only, useful recovery of motor and sensory functions in 7 out of 10 patients with total plexus lesions. Regarding these good results of operative treatment of mechanical lesions of the peripheral nerves an exact clinical and neurophysiological examination has to be done for diagnosing and localizing those peripheral nerve affections. PMID- 2992171 TI - Paraneoplastic syndromes of the nervous system. AB - The paraneoplastic disorders of the nervous system are clinicopathologic syndromes that occur in association with systemic cancer. Dysfunction of any part of the nervous system or its effector organs may be produced by a paraneoplastic process. The temporal relationship between the onset of a paraneoplastic syndrome and the diagnosis of the associate cancer varies. Most of the paraneoplastic syndromes of the nervous system may appear months to years before the cancer is discovered. The pathogenesis of most of the paraneoplastic syndromes of the nervous system is unknown and specific, effective therapy is lacking. With rare exceptions, treatment of the associated cancer seldom alters the course of patients with these syndromes. However, in other paraneoplastic syndromes such as myasthenia gravis, myasthenic syndrome of Eaton-Lambert and polymyositis dermatomyositis an autoimmune pathogenesis is established or strongly suspected. PMID- 2992174 TI - Identification of hepatic metabolites of n-heptyl-delta-1-tetrahydrocannabinol in the mouse. AB - Nineteen metabolites of heptyl-delta-1-THC have been identified in mouse liver using combined g.l.c. and mass spectrometry following i.p. administration of the drug. All metabolites contained either hydroxyl or carboxylic acid functions, the positions hydroxylated (2'', 3'', 6 and 7) were the same as those found previously for delta-1-THC in this species. The relative abundance of acid metabolites was much lower than for the corresponding metabolites produced from delta-1-THC itself, but hydroxylation was more extensive with metabolites containing up to four hydroxyl groups being observed. PMID- 2992175 TI - Stereospecific hydrolysis of 8 alpha,9 alpha- and 8 beta,9 beta epoxyhexahydrocannabinols in the mouse in vivo and in vitro. AB - After i.v. injections to the mouse, both 8 alpha,9 alpha- and 8 beta,9 beta epoxyhexahydrocannabinols (EHHCs) were easily hydrolysed to 8 beta,9 alpha dihydroxyhexahydrocannabinol (dihydroxy-HHC) rather than 8 alpha,9 beta-dihydroxy HHC in the liver. 8,9-Dihydroxy-HHCs hydroxylated at the 2' or 3' position of a pentyl side-chain were identified in vivo as liver metabolites of the epoxides. The same stereospecific hydrolysis of 8,9-EHHCs were observed in vitro using liver-microsomal fractions of mice, though both epoxides were resistant to enzymic hydrolysis. Several monohydroxylated 8,9-EHHCs, together with small amounts of 8,9-dihydroxy-HHCs were identified as in vitro liver metabolites of the epoxides. These metabolites could be formed not only by epoxide hydrolase but also by a mono-oxygenase system involving cytochrome P-450. The magnitude of the binding affinities of cannabinoids to cytochrome P-450 was ranked in the following order: delta 8-tetrahydrocannabinol (spectral dissociation constant Ks = 9.4 microM) greater than 8 beta,9 beta-EHHC (13.3 microM) greater than 8 alpha,9 alpha-EHHC (34.5 microM). PMID- 2992177 TI - [Malignant eccrine poroma]. AB - We report on a 82-year-old woman suffering from malignant eccrine poroma on her back. The infiltrated surrounding tissue suggested lymphangiosis carcinomatosa; one axillary lymph node was enlarged. Both features, however, turned out to be only of inflammatory nature. PMID- 2992176 TI - [Generalized verrucosis in familial T-helper cell deficiency. Treatment with interferon and Tigason]. AB - We report on a 17-year-old patient suffering from generalized verrucosis. This diagnose had been confirmed by means of light microscopy, electron microscopy, and DNA hybridisation (HPV type 2). The study of immunological parameters revealed reduced numbers of T-lymphocytes, especially of helper-T-cells. Skin tests with recall antigens were normal. Three brothers of our patient showed slight reduction of helper-T-cells. Treatment with topical retinoic acid was not effective. Treatment with alpha-interferon i.m. intralesionally was effective in the beginning, but two weeks after onset of therapy no further improvement could be achieved. Discontinuation of therapy was followed by a relapse. Tigason which was given thereafter led to flattening of the warts but failed to bring about complete regression within the following four months. PMID- 2992178 TI - [Determination of cAMP concentrations in the liver of the rat using the luciferin luciferase method]. PMID- 2992179 TI - Proton uptake by light induced interaction between rhodopsin and G-protein. AB - The light-induced proton uptake of rod outer segment disc membranes has been investigated in the absence and presence of G-protein. Proton uptake was measured as the alkalisation of the suspending medium using a pH electrode and/or the indicator dye bromocresol purple. It was found that besides the known proton uptake of photolysed rhodopsin additional uptake of one proton accompanies formation of the complex between rhodopsin and G-protein. No measurable proton uptake was found under conditions of rapid redissociation of the complex indicating an only transient protonation during its lifetime. Proton uptake was the same in washed membranes recombined with G-protein and in ordinarily stacked rod outer segments. The additional proton uptake reported here is not due to enhanced formation of the protonated photoproduct metarhodopsin II. PMID- 2992180 TI - Fractionation of Plasmodium chabaudi-infected erythrocytes into parasites and ghosts. AB - We have developed a new procedure for subfractionation of Plasmodium chabaudi infected erythrocytes into parasites and ghosts. Trophozoite-infected erythrocytes enriched over a percoll-step (:1.10 g/cm3) are subjected to a glycerol-enhanced osmotic shock. This induces the release of parasites and the emergence of erythrocyte ghosts, which can be separated on a continuous percoll gradient (:1.02----1.10 g/cm3). The parasites are intact in terms of ultrastructure and incorporation of 14C-isoleucine. The erythrocyte ghosts are purified over a two-step percoll gradient (:1.01-1.02 g/cm3). Our method recovers about 40%-50% of the initial ghosts and 70%-95% of the initially freed parasites. PMID- 2992181 TI - [Hemodynamics, various aspects of aldosterone metabolism and renin activity in children with cerebral palsy]. PMID- 2992182 TI - [Pathology of organs of the urinary system in children with high risk of prenatal Coxsackie virus infection]. PMID- 2992184 TI - Isolation and biochemical characterization of intertypic recombinants of foot-and mouth disease virus. AB - Recombinants were isolated between two European serotypes (O and A) and between two of the most distantly related serotypes (O from Europe and SAT2 from Africa) using appropriate ts mutants in an infectious centre assay. The recombinants were characterised by electrofocusing of their induced proteins and by RNase-T1 fingerprinting of their RNA. The approximate location of the cross-over event in each recombinant was determined by sequencing the unique distinguishable O or A oligonucleotides and locating them within the known genome sequence. Nine different types of recombinant were identified from the two types of cross (O X A and O X SAT) and all had a single cross-over in the middle or 3' half of the genome, i.e. in the nonstructural coding region. Recombination between the most distantly related viruses (O X SAT2) appeared to occur at a lower frequency than recombination between serotypes of the same group (O X A). A higher incidence of recombinant proteins with unique pI was also observed in the O X SAT2 crosses. PMID- 2992183 TI - Characterization of leader-related small RNAs in coronavirus-infected cells: further evidence for leader-primed mechanism of transcription. AB - Mouse hepatitis virus (MHV), a murine coronavirus, replicates in the cytoplasm and synthesizes 7 viral mRNAs containing an identical stretch of leader RNA sequences at the 5'-end of each RNA. The leader-coding sequences at the 5'-end of genomic RNA are at least 72 nucleotides in length and are joined to the viral mRNAs by a unique mechanism. Utilizing a leader-specific cDNA probe, we have detected several free leader RNA species ranging from 70 to 82 nucleotides in length. The predominant leader RNA was approximately 75 nucleotides. In addition, larger distinct leader-containing RNAs were also detected ranging from 130 to 250 nucleotides in length. The 70-82-nucleotide leader-related RNAs were present in both the cytosol and membrane fractions of infected cells. They were also detected only in the small RNA fractions but not associated with the replicative intermediate RNA. These data suggest that the leader RNAs were associated with the membrane-bound transcription complex but at least part of them were dissociated from the RNA template. We have also identified a temperature sensitive mutant, which synthesizes only leader RNA but not mRNAs at nonpermissive temperature, indicating that leader RNA synthesis is distinct from the transcription of mRNAs. These data support the leader-primed mechanism for coronavirus transcription and suggest that one or more free leader RNAs are used as primers of mRNA synthesis. PMID- 2992185 TI - [Effect of 8-Br-cAMP on the formation of conditioned reflex behavior in the white rat]. AB - The influence of cAMP analogue 8-Br-cAMP on conditioning was studied in white rats. Two models of learning were used with different kinds of reinforcement, i. e. conditioned active avoidance and instrumental alimentary reactions in a complex maze. Intraventricular 8-Br-cAMP injection 4 or 24 hours before the beginning of learning improved the process of defensive as well as alimentary conditioning. Characteristics of formation of complex behaviour of experimental rats in a maze showed that under the influence of 8-Br-cAMP, not only conditioning was accelerated, but the process of optimal decision making itself was changed. The data obtained permit to suppose that 8-Br-cAMP first of all affects initially poorly learning rats. PMID- 2992186 TI - [Effect of enkephalins on the plasticity of pond snail]. AB - The effects of introduction of leu- and met-enkephalins (LE and ME) into the perfusion medium on the characteristics of electrical activity, spontaneous firing and habituation of single neurones to repeated intracellular electrical stimulation were studied on isolated CNS of molluscs. The character, speed, and degree of development of endoneuronal habituation changed significantly after application of LE in 67 per cent of the neurones studied and in 25 per cent of neurones after ME application. As a rule the changes of habituation dynamics occurred at constant levels of membrane potential, excitability, and reactivity of the neurone. LE and ME exerted different modulating effects on the initial electrical activity in 60 per cent of neurones. The obtained data on the independence of the effects of enkephalins on different parameters of activity of one and the same neurone give evidence of a mosaic character of excitable neuronal membrane. A suggestion is made about the possible role of the revealed opiate dependence of the endoneuronal functional plasticity in realization of opiate influences at the behavioural level. PMID- 2992187 TI - [Possible role of hypoxia and lipid peroxidation in the development of a neurosis like condition in the rat]. AB - The chronic emotional pain stress resulting in a development of neurosis-like state in rats induced an increase of arterial pressure and change of the cardiac rate dynamics under the conditions of functional load. An increase of cardiac mass was also seen without change of masses of the thymus, adrenal glands and the spleen. The rise of activity of cytochromeoxidase and activation of peroxide lipide oxidation (by malonate dialdehyde level) were observed in the cerebral cortex and the hippocampus of neurotized rats. Injection of antioxidant F-801 before each emotional pain stress trial prevented vegetative disturbances, cardiac hypertrophy, and increase of oxidative activity in the brain. The role of peroxide lipide oxidation and that of the factor of hypoxia in development of disturbances caused by neurotization were discussed. PMID- 2992188 TI - [Enzyme immunochemical detection of the use of hashish and confirmation by thin layer chromatography]. AB - The EMIT cannabinoid assay was used for screening blood and urine after smoking tetrahydrocannabinol (THC; 10 mg) or ingestion of THC (30 mg). Cannabinoid levels in urine remain detectable up to 1 week. Confirmation was done by adsorption of the THC carboxylic acid onto a C18 extraction column and elution with acetone and TLC. The method is simple and sensitive and is applicable with common laboratory equipment. The detection limit is 10 ng/ml, using 10 ml urine. PMID- 2992190 TI - [Carcinoma of the appendix arising on the basis of a mucinous cystadenoma]. AB - The author describes a rare case of mucinous adenocarcinoma arising in a mucinous cystadenoma of the appendix. The tumor developed into a mucocele which subsequently perforated. At a second operation two years later a malignant pseudomyxoma peritonei was found. PMID- 2992189 TI - [Death by peroral ingestion of soluble glass (sodium silicate)]. AB - The intake of 0.51 of water glass (sodium metasilicate; colloid pH 12.5) led to death within 1-1.5 h. Autopsy revealed alkali burns of the stomach mucosa. The stomach contained a small amount of liquid with a pH of 11.5. Histologically, numerous bronchioles and alveoles were filled with amorphous material. This material was chemically proved to be condensed water glass. The obstruction of large parts of the lungs by water glass led to an inhibition of alveolar gas diffusion, which may have been the cause of death. The transformation of the water glass from liquid to solid occurred in the lungs by means of the carbonic acid of expiration air. This was made possible because the pH of water glass had been changed by gastric secretions from 12.5 to 11.5. Water glass starts to solidify around pH 11.4-11.3. PMID- 2992191 TI - Preliminary evaluation of the ELISA as a tool for the detection of rotaviruses in activated sewage sludge. AB - Samples of activated sludge were taken in three different treatment plants near Nancy (France) and in West Berlin (F.R.G.), and were investigated for their content of rotaviruses. After elution with beef extract (0.25%, pH 7.3) and ultrasonication, the samples were concentrated by protamine flocculation and, in a few cases, in parallel by ultracentrifugation. A commercially available ELISA test kit (Rotazyme, Abbott Diagnostics) was routinely used for rotavirus detection. Although both the Rotazyme and a laboratory-designed ELISA gave positive results for the suldge concentrates, confirmation with a blocking test and electron microscope (EM) was negative in all cases so far investigated. Therefore, the exclusive use of ELISA cannot be recommended for similar investigations as it may lead to false positive results. In this light, an additional verification step with neutralizing antibodies should be included by the ELISA kits' manufacturers to enable a quick confirmation of the user's results. PMID- 2992193 TI - Drugs recently released in Belgium. Nabilone and rosoxacin. PMID- 2992192 TI - [Various rarely encountered adverse effects and complications of psychopharmacotherapy]. AB - Nine patients are described in whom psychopharmacotherapy was associated with the development of rare complications in the form of the syndrome of cortical disturbances, psychosensory disorders and toxic polyneuropathy. All patients presented residual organic insufficiency of the CNS, with some of them exhibiting endocrine-metabolic disorders as well. PMID- 2992194 TI - Viral disease as a complication of blood transfusion. PMID- 2992195 TI - The receptor concept revisited. PMID- 2992196 TI - Fine needle aspiration biopsy cytology of major salivary glands. AB - Fine needle aspiration biopsy of the major salivary glands was performed on 160 patients. In 146 patients with satisfactory samples, the cytologic diagnosis was correlated with clinical follow-up and histologic findings. There were 122 benign lesions, including 47 tumors. There were 24 malignant lesions, 10 of which were primary and 14 metastatic. The overall accuracy was 98%. The sensitivity of the technique was 87.5%. There was no false-positive diagnosis. There were three false-negative diagnoses due to sampling errors and inexperience during the initial period of the study. This study documents that needle aspiration biopsy cytology of the salivary glands is accurate, simple, rapid, inexpensive, well tolerated and harmless to the patient. PMID- 2992197 TI - Adenoid cystic carcinoma arising in a pleomorphic adenoma of the parotid gland. An aspiration cytology and ultrastructural study. AB - The unusual occurrence of an adenoid cystic carcinoma arising in a pleomorphic adenoma is described, with emphasis on the cytomorphology of the neoplasm in a fine needle aspiration specimen. The ultrastructural findings are detailed, including the acellular hyaline spheres so characteristic of this tumor. PMID- 2992198 TI - Fine needle aspiration of adenoid cystic carcinoma metastatic to the lung. Cytologic features and differential diagnosis. AB - Two examples of adenoid cystic carcinoma metastatic to the lung, one from a Bartholin's gland and the other from a submandibular gland, were sampled by fine needle aspiration. Although the cytologic features of adenoid cystic carcinoma have been well described, it is easy to confuse adenoid cystic carcinoma with other more common primary small-cell neoplasms of the lung; to determine distinguishing features, we compared the cytomorphology of adenoid cystic carcinoma with well-differentiated adenocarcinoma, small-cell undifferentiated carcinoma and carcinoid tumor of the lung. The differential features distinguishing adenoid cystic carcinoma from these other neoplasms include: (1) tight, globular, honey-comb arrangements of cells lacking true nuclear molding; (2) acellular chunks of basal lamina material, which alone may suggest adenoid carcinoma; and (3) the extension of a solid core of basal lamina material beyond a sievelike cellular meshwork. The morphologic expression of metastatic adenoid cystic carcinoma is so distinctive as to permit a definite diagnosis. PMID- 2992199 TI - Adenoid cystic carcinoma metastatic to the lung. Report of a case diagnosed by fine needle aspiration biopsy cytology. AB - Cytologic examination of a fine needle aspiration (FNA) biopsy specimen served to diagnose a lung metastasis in a patient with an adenoid cystic carcinoma primary in a minor salivary gland of the hard palate. The aspiration cytologic features of the metastatic adenoid cystic carcinoma were similar to those of primary adenoid cystic carcinoma, including uniform tumor cells with scant cytoplasm surrounding transparent, mucoid spherical bodies. FNA biopsy cytologic diagnosis of metastatic adenoid cystic carcinoma is accurate, particularly if mucoid spherical bodies are present. PMID- 2992200 TI - Malignant islet-cell tumor of the pancreas diagnosed by fine needle aspiration biopsy. A case report. AB - A case of malignant islet-cell tumor of the pancreas diagnosed by preoperative percutaneous fine needle aspiration biopsy and confirmed by tissue examination is reported. The cytologic presentation of this neoplasm is described, and the differential diagnosis is discussed. PMID- 2992202 TI - Granular-cell tumor of the breast. A cytologic, immunohistochemical and ultrastructural study of two cases. AB - Two primary granular-cell tumors of the breast are reported. Cytologically and histologically, these tumors had the classic features of small, eccentrically located nuclei and numerous periodic acid-Schiff-positive cytoplasmic granules. The characteristic cytologic features were best appreciated in touch imprints, not in frozen sections. Immunohistochemically, the tumors demonstrated diffuse cytoplasmic staining for S-100 protein but negative staining for myoglobin. The significance of these immunohistochemical staining characteristics, particularly in evaluating the possible histogenesis of these tumors, is discussed. The ultrastructural features of these two tumors are also presented and compared to findings reported by other investigators. PMID- 2992201 TI - Aspiration and exfoliative cytology, including ultrastructure, of a malignant granular-cell tumor. AB - The cytopathologic features of a malignant granular cell tumor in both exfoliative and aspiration cytopreparations included isolated cells with low nuclear-cytoplasmic ratios and abundant, diffusely granular cytoplasm. Transmission electron microscopy revealed the diagnostic secondary lysosomes in the neoplastic cells. PMID- 2992203 TI - Cytologic findings in signet-ring adenocarcinoma of the large bowel. A case report. AB - Signet-ring adenocarcinoma of the large bowel is an uncommon tumor with distinctive gross anatomic, radiologic and histologic features. A case in a 34 year-old man is presented. Because of the unusual appearance of the bowel at colonoscopy, diagnostic cytologic smears were taken. The patient subsequently underwent resection of this tumor but died within 20 months of metastatic disease. The histologic and cytologic features are described; the cytologic findings do not appear to have been previously reported. PMID- 2992204 TI - Pulmonary oncocytoma. Report of a case with cytologic, histologic and electron microscopic study. AB - The cytology of a pulmonary oncocytoma diagnosed by bronchial brushings is described along with the light and electron microscopic findings. Ultrastructurally, the tumor cells showed mitochondrial hyperplasia and an absence of neurosecretory granules. The possible histogenesis of pulmonary oncocytomas is discussed. PMID- 2992205 TI - Secretory, enzymatic, and morphological characterization of rat pancreatic endocrine tumours induced by streptozotocin and nicotinamide. AB - Rat pancreatic endocrine tumours were induced by administration of streptozotocin plus nicotinamide. Fifteen to eighteen months later tumours with wet weights of 0.1 to 224 mg were isolated. These tumours were compared with normal rat pancreatic islets. Insulin release from perifused tumours was stimulated by D glucose, L-leucine, 2-ketoisocaproate, and D-glyceraldehyde, potentiated by theophylline and inhibited by norepinephrine. Compared with isolated rat pancreatic islets, however, insulin secretory responsiveness to glucose stimulation and insulin content were reduced in tumour tissue. Hypoglycaemia in tumour bearing rats and impaired diffusion of insulin out of the tumours may explain this difference. The pattern of enzyme activities observed in tumour tissue was typical for pancreatic endocrine tissue. The activities of succinate dehydrogenase, the two types of the monoamine oxidase, and alpha-glucosidase were in the normal range in tumour tissue. Only the activities of 5'nucleotidase and glutamate dehydrogenase were decreased. Immunocytochemical analysis of the tumours revealed that they contained an average of 91% B-cells. In addition 8% of D-cells were encountered. Proportions of A-cells and PP-cells ranged below 1%. Thus this endocrine tumour of the pancreas with a high proportion of functionally intact B-cells is an interesting model for studying regulation of secretion and endocrine tumour development. PMID- 2992206 TI - Multiple-sites of inhibition by intravenous metyrapone of human adrenal steroidogenesis. AB - The in vivo influence of metyrapone on adrenal steroidogenesis has been studied by measuring plasma concentrations of pregnenolone, 17-OH-pregnenolone, progesterone, 17-OH-progesterone, 11-deoxycorticosterone, 11-deoxycortisol, corticosterone, cortisol, 18-OH-11-deoxycorticosterone, 18-OH-corticosterone and aldosterone before, during and after a 5 h infusion of metyrapone ditartrate at doses of 0.2 g/h and 0.8 g/h respectively. Time courses of plasma steroids and corticotrophin indicate an inhibitory effect of metyrapone on total adrenal steroidogenesis in addition to the known inhibition of the 11- and 18 monooxygenase. The effect on total adrenal steroidogenesis is pronounced at high concentrations of metyrapone and may be compensated by corticotrophin. This effect and a concomitant suppressive effect of metyrapone on plasma corticotrophin itself may account for the frequently described falsely abnormal responses to the metyrapone test. From the present 'in vivo' data, no significant, metyrapone induced alterations of the 3 beta-hydroxysteroid dehydrogenase/delta 5-isomerase, 17-monooxygenase, 21-monooxygenase or the 18 hydroxysteroid dehydrogenase are apparent. PMID- 2992207 TI - 'Cryptic' form of congenital adrenal hyperplasia due to 21-hydroxylase deficiency in the Yugoslav population. AB - Five individuals with the asymptomatic, 'non-classical', 'cryptic' form of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase (21-OH) deficiency from 5 unrelated families were discovered during hormonal studies and HLA-typing performed in a series of 24 families with CAH due to 21-OH deficiency. Four of the 5 individuals with the 'cryptic' form of CAH belong to families where the index case was a patient with the classical form of CAH due to 21-OH deficiency. The fifth one originated from a family where the index case was a girl with the 'non-classical', 'late-onset' form of the disease. All the 5 individuals had no clinical symptoms in spite of clearcut biochemical signs of 21-OH deficiency: increased 17-OH-progesterone (17-OHP), dehydroepiandrosterone and androstenedione levels, particularly after ACTH-stimulation. The 17-OHP response upon ACTH stimulation of heterozygotes for this 'non-classical' form of 21-OH deficiency did not differ from the response of heterozygous individuals for the classical form of the disease. The results of this study confirm the hypothesis that individuals with the 'cryptic' form of CAH due to 21-OH deficiency are genetic compounds bearing one allele for the severe, classical form, and on the homologous locus, another one for the mild 'non-classical' form of CAH due to 21 OH deficiency. Their genotype was 21-OH severe/21-OH mild.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992208 TI - Hormonal regulation of ascorbic acid in the adrenal of the rat. AB - Fifteen days after hypophysectomy of rats the concentrations of ascorbic acid (AA) in adrenals, liver, blood and urine are lower than in normal rats or in rats 1 day after hypophysectomy. Despite the low levels the percentage AA depletion after administration of ACTH and the subsequent repletion to the pre-ACTH level are normal. Lack of corticosteroids is not the cause of the low AA levels, as shown by experiments in 15 days adrenalectomized rats and in rats treated with low doses of dexamethasone. Fifteen days treatment with high doses of dexamethasone lowered the AA concentrations in adrenals, liver and blood. Treatment with long-acting ACTH maintained adrenal weight but not adrenal and blood AA. A high dose of ACTH lowered these levels. The administration of AA markedly increased the AA levels in blood, but did not normalize its concentration in the adrenals, not even when the size of the adrenals was maintained by treatment with long-acting ACTH. Growth hormone, in particular when administered together with long-acting ACTH, markedly raised the AA concentration in the adrenals but hardly affected the AA blood level. Rats with high blood levels of prolactin induced by pituitary grafts in the kidney also had clearly higher AA levels in adrenals, but not in blood. These results indicate that, although acute AA release from the adrenal is caused by ACTH, AA uptake to a certain concentration is not controlled by the pituitary gland, and above this concentration is promoted by growth hormone and prolactin. In the liver AA release may also be caused by ACTH but the AA production is promoted by other as yet unidentified pituitary factors. PMID- 2992209 TI - Calcium metabolism in paravertebral ligamentous ossification. AB - Twenty-eight patients with paravertebral ligamentous ossification (PVLO) and 11 control subjects were studied in an attempt to examine possible involvement of disturbances in mineral metabolism in the development of PVLO. No significant difference in baseline serum calcium and fasting urinary calcium excretion was found between patients with PVLO and controls. Patients with PVLO showed lower serum inorganic phosphate (Pi) and tubular reabsorptive capacity for Pi (TmP/GFR) than controls. Basal nephrogenous cyclic 3',5'-AMP (NcAMP) was elevated in some patients with PVLO. A significant inverse relationship was found between basal TmP/GFR and NcAMP in patients with PVLO (r = -0.50; P less than 0.01). Compared with controls, patients with PVLO had significantly lower calciuric responses to an oral calcium load (P less than 0.05), suggesting decreased intestinal calcium absorption. Furthermore, a significant inverse correlation was found between basal NcAMP and the calciuric response in patients with PVLO (r = -0.42; P less than 0.05). Serum 25-hydroxyvitamin D (25OHD) and 1,25-dihydroxyvitamin D (1,25(OH)2D) levels were not significantly different between the two groups. The size of ossified areas in paravertebral ligaments estimated from spinal radiograms inversely correlated with the calciuric response to the oral calcium load (r = -0.50; P less than 0.01). These data demonstrate that the development of PVLO is associated with decreased intestinal calcium absorption in the face of normal serum 1,25(OH)2D level. Thus, together with previous observations showing a high incidence of PVLO in patients with hypoparathyroidism or familial hypophosphataemic rickets/osteomalacia, the present results suggest that the defect in the action of 1,25(OH)2D may underlie the development of PVLO. PMID- 2992210 TI - [Education project of leprosy teams in health education. Ministry of Public Health and Population, Republic of the Ivory Coast]. PMID- 2992211 TI - Tangier disease. A case with sensorimotor distal polyneuropathy and lipid accumulation in striated muscle and vasa nervorum. AB - A 65-year-old man with Tangier disease (analphalipoproteinemia) had had a progressive sensorimotor distal neuropathy with sensory ataxia for 1 year. Muscle biopsy demonstrated excess lipid vacuoles on histochemical and electron microscopic techniques. Sural nerve biopsy showed a marked loss of large fibers and an increase in small myelinated fibers, with presence of remyelinating fibers and clusters of regeneration; a few aspects of active demyelination and some onion-like formations were also present. Lipid accumulation chiefly affected the Schwann cells of unmyelinated fibers and, to a lesser degree, of myelinated fibers, endoneurial fibroblast, and vasa nervorum. Teased fibers showed prevalent aspects of de-/remyelination and, often in association, marked myelin wrinkling suggesting axonal atrophy. This Tangier patient differs from known cases for the presence of a distal symmetrical sensorimotor polyneuropathy (not previously reported in Tangier disease) and because of the morphological findings of de /remyelination coexisting with aspects of axonal atrophy and previous degeneration, and of lipid accumulation within striated muscle and vasa nervorum. This latter finding contrasts with the assumption that in Tangier disease vessel walls are not a site of lipid storage: probably the vasa nervorum are different, in this respect, from other vessels, because of the intense lipid metabolism of the nervous tissue. Thus we suggest that involvement of vasa nervorum in Tangier disease may be more important than previously suspected, possibly playing a role in the causation of neuropathy. PMID- 2992212 TI - Immunocytochemical demonstration of S-phase cells by anti-bromodeoxyuridine monoclonal antibody in human brain tumor tissues. AB - Five patients with various brain tumors received bromodeoxyuridine (BrdU), 150 200 mg/m2 i.v., at the time of craniotomy. Biopsied materials were fixed in 70% ethanol, sectioned, denatured with hydrochloric acid, and reacted with monoclonal antibodies against BrdU. Immunofluorescence and immunocytochemical methods were used to visualize BrdU-labeled nuclei. Our results showed that both methods demonstrated BrdU-labeled nuclei satisfactorily in tissue sections. Thus, BrdU can be used to measure the proliferative potential of human tumors in situ. PMID- 2992213 TI - Nuclear inclusions following chronic ethanol administration. An electron microscopic investigation of the rat parietal cortex. AB - Wistar rats with an average initial body weight of 180 g received a 15% (v/v) ethanol-water solution over a period of 6 months. Neurons of the parietal cortex (area 3) showed a significant increase of nuclear inclusions which consisted of parallel filaments and tubular systems. Furthermore, we observed an augmentation of lipid droplets, nemetosomes, and straight filaments. The latter could not be identified in control animals. It is discussed whether the occurrence of these structures might be enhanced by an altered cellular metabolic activity during the chronic ethanol administration. PMID- 2992214 TI - The nature of cytoplasmic inclusions in cerebellar haemangioblastomas. AB - Intracytoplasmic hyaline globules present in stromal cells in eight of a series of ten cerebellar haemangioblastomas have been shown to contain the glycoprotein alpha-1-antitrypsin. PMID- 2992216 TI - Pathology of intraocular lens implantation. Paulo Foundation International Symposium, 18 May, 1984, Helsinki, Finland. PMID- 2992215 TI - Effects of clonidine on polypeptide and steroid hormone levels in man. A review. PMID- 2992218 TI - Pseudophaco--anaphylactic endophthalmitis? P.M.M.A. related. AB - The pathological findings of pseudophacoanaphylactic endophthalmitis are described in a patient with a combined trabeculectomy, extracapsular cataract extraction and an "in-the-bag" posterior chamber lens implant, and who developed sympathetic ophthalmitis following a perforating eye injury within hours of his operation. Apart from the typical sympathetic histological findings, the eye revealed a pseudophaco-anaphylactic endophthalmitis reaction which appeared to be totally related to the polymethylmethacrylate (P.M.M.A.) optic. The "in-the-bag" polypropylene closed loops revealed no significant histological reaction. PMID- 2992217 TI - Histopathologic study of changes occurring in eyes with intraocular lens implantation: autopsy eyes, enucleated eyes and corneal buttons. AB - The clinicopathologic findings in 17 eyes which were surgically enucleated or removed at autopsy as well as the histopathology of 72 corneal buttons removed from eyes with intraocular lenses are described. In four of five autopsy eyes the intraocular lens was well tolerated; however, in all cases, changes related to the pseudophakos were found. In seven of 12 surgically enucleated eyes, postsurgical bacterial or fungal endophthalmitis had developed. On histopathological examination, one eye revealed clinically unsuspected endophthalmitis phacoanaphylactica after extracapsular cataract extraction and intraocular lens implantation. In two cases, choroidal melanoma became evident after cataract extraction and insertion of a pseudophakos. In both cases preoperative ultrasonography was limited to A-scan axial measurements. The 72 corneal buttons were removed from patients with pseudophakic bullous keratopathy; histologically the most striking findings were guttata-like changes in 50% of the specimens and retrocorneal membranes in 13.9% of the specimens. PMID- 2992219 TI - Successful intraocular lens implantation: a histological and scanning electron microscopic study of four cases. AB - The histological findings in four globes obtained at autopsy from clinically uncomplicated cases with intraocular lens implants, one glass posterior chamber and three anterior chamber lenses consisting of two pupillary fixated and one "in the-bag" fixated Binkhorst lens, are reported. Minimal changes could be detected in the tissues with which the implants were in contact. Scanning electron microscopy of the cornea in the three anterior chamber implants showed a loss of endothelial cells in the axial region of roughly 30-50%. There was marked vertical disparity in the loss. Scanning microscopy of the implants showed evidence of surface degradation of the nylon loops of all the anterior chamber implants. PMID- 2992220 TI - Histopathological changes in eyes following intraocular lens implantation (IOL). Preliminary study of a Danish and Swedish material. PMID- 2992221 TI - Cellular deposits on intraocular lenses. AB - We examined cellular deposits on 14 intraocular lenses (IOL) that were removed after implantation for a variety of clinical reasons. Five of the lenses were cultured to evaluate the growth potential of preexisting cells on the IOLs. The most common cells found were iris pigment epithelial cells. The thickest membranes were probably formed by iris stromal cells. Other cellular deposits included macrophages, multinucleated foreign body giant cells, lens epithelial cells, squamous epithelial ingrowth, red blood cells, uveal melanocytes, and vitreous strands. Cells around the IOL had only limited growth in tissue culture. We propose that a continuous influx of inflammatory cells in conjunction with a disruption of the blood-aqueous barrier may be an important pathogenetic mechanism for the production of thick secondary membranes around an IOL. PMID- 2992222 TI - Posterior chamber lens implantation. A clinicopathological report of a successful case. AB - This report describes the clinical and histopathological studies of an eye 13 months after successful extracapsular cataract surgery and J-loop-type posterior chamber lens implantation. The lens was of PMMA while the loops were made of polypropylene. The corrected visual acuity of the patient was 1.0 and clinically the eye showed no evidence of anterior uveitis. The lower loop was in the capsular bag while the upper loop was embedded in the ciliary body surrounded by chronic foreign body giant-cell inflammatory reaction. PMID- 2992223 TI - Microscopic-cytologic investigations on explanted intraocular lenses. AB - Nine iris supported anterior chamber lenses were removed during keratoplasty, eight of them on account of bullous pseudophakic keratopathy. They were examined cytologically according to the method of WOLTER. No one revealed a complete cellular--membraneous capsule. This membrane has with great probability a protective function. Four basic types of cells could be characterized. All cells of the borders of the eye chambers are involved but predominantly mobile and sessile macrophages are the source of the covering cells. The cellularity depends on the type of the lens and the material of the lens and on the clinical picture of the disease. Supramid is worse than prolene. Dangerous are pseudophakodonesis and endothelial contact. PMID- 2992224 TI - Cochlear processes reflected in responses of the cochlear nerve. AB - The effects of sound frequency, intensity, and duration on responses of cochlear nerve fibers and inner hair cells (IHCs) are reviewed and compared. The frequency selectivity observed in the nerve is already present in the IHC receptor potential but synaptic transmission appears to influence some other properties of the nerve response. First, the average rate-intensity function of a nerve fiber spans a smaller operating range than the IHC input-output characteristic. However, nerve fibers with different sensitivities can innervate the same IHC and respond over different regions of its characteristic. Second, adaptation is present in the nerve-fiber response but not observed in the IHC. It appears to result from a decrease in synaptic transmission and produces temporal contrast and a decrease in operating range. Interactions among the various effects have important influences on the spatiotemporal pattern of cochlear-nerve responses to complex stimuli. PMID- 2992225 TI - Development of B lymphocyte function in childhood. AB - The capacity of blood lymphocytes of children aged from birth to six years to produce immunoglobulins was studied in vitro at the cell level using a direct B lymphocyte activator (Epstein-Barr virus) or a T lymphocyte dependent B lymphocyte activator (pokeweed mitogen). Umbilical cord blood lymphocytes secreted IgM at adult levels after Epstein-Barr virus stimulation, while the ability to synthesize IgG and IgA increased up to the ages of 1 and 2 years, respectively, but not beyond this period. IgG3 production preceded that of the other IgG subclasses. The T lymphocyte dependent IgM synthesis was low at birth, but approached adult levels at two years of age. T cell dependent IgG and IgA secretion, however, remained reduced even up to 6 years of age. PMID- 2992226 TI - A 16-month-old boy with infectious mononucleosis, parotitis and Bell's palsy. AB - The case of a 16-month-old boy with infectious mononucleosis who developed bilateral parotitis and unilateral Bell's palsy is presented. The diagnosis was confirmed by the development of specific antibodies for various Epstein-Barr virus antigens typical of primary Epstein-Barr virus infection. Serological tests for other viruses were negative. The Bell's palsy subsided within three months, while the clinical signs of infectious mononucleosis and parotitis persisted for three weeks. PMID- 2992227 TI - Lipid storage disease: Part II. Ultrastructural pathology of lipid storage cells in sphingolipidoses. AB - The ultrastructural pathology of the stored materials in lipid storage cells, particularly of macrophagic nature, in various disorders of sphingolipidosis was investigated. Cell morphology of the lipid storage cells was largely divided into two groups; one had peculiar cell morphology, such as Gaucher cells or globoid cells, and the other showed the appearance of foam cells. These cytological characteristics of the lipid storage cells were closely related to the ultrastructural configuration of lipid storage inclusions. By transmission electron microscopy, the fundamental structures of the stored materials were classified into two types; tubular and lamellar. The tubular structures were formed by accumulation of ceramide or monohexosyl ceramide, whereas the lamellar structures were formed by accumulation of larger sphingolipids than monohexosyl ceramide. These tubular structures were proven to consist of multilayers of lamellae, which are considered fundamentally similar to the lamellar structures. Almost all the lipid storage inclusions are considered to be of lysosomal origin, because of their encirclement by a single unit membrane and localization of acid phosphatase activity, and participation of heterophagic or autophagic mechanisms as for the development of the inclusions may be noted. Besides, the occurrence of secondary lipid storage was pointed out in some disorders of sphingolipidosis. PMID- 2992228 TI - Myoepithelioma of the parotid gland. Report of two cases with immunohistochemical technique for S-100 protein and electron microscopic observation. AB - Two cases of myoepithelioma occurring in the parotid gland were reported. These tumors consisted of spindle cells that exhibited an arrangement similar to leiomyoma or fibroma. The diagnosis was made on the basis of characteristic ultrastructural as well as immunohistochemical findings: The tumor cells of the two cases likewise contained myofilaments and gave a positive result for the S 100 protein immunohistochemical technique. PMID- 2992230 TI - Congenital nephrotic syndrome (Finnish type). AB - The case was a one year and four months old boy, who had been diagnosed as congenital nephrotic syndrome on the 4th day of life. He died of septicemia from peritonitis. Large pale kidneys, dilated trunk of pulmonary artery, and thickened left atrial endocardium were observed at autopsy. Renal histology seemed to be compatible with congenital nephrotic syndrome of the Finnish type. Viral antigens of herpes simplex type 1 and varicella-zoster in paraffin-embedded sections were proven to be negative. Besides renal change, increased number of pancreatic islets without hypertrophy was noticed. PMID- 2992229 TI - Nonchromaffin paraganglioma in the nose and paranasal sinuses. AB - Nonchromaffin paraganglioma occurring in the nose and paranasal sinuses are extremely rare. The authors have experienced a case of nonchromaffin paraganglioma of nasal cavity which extended to the epipharynx. To our knowledge, this reported case is the fourth case that has appeared in the world literature. The patient, a 31-year-old female, had complained of a right nasal obstruction and mucopurulent rhinorrhea for two years. The right nasal cavity was filled with a polypoid mass showing no bleeding tendency or necrosis. Radiographs revealed a homogeneous shadow which occupied the entire nasal cavity and paranasal sinuses. Histopathological studies revealed that the tumor showed typical findings of nonchromaffin paraganglioma and neurosecretory granules in cytoplasm were observed by electron microscopic study. Subsequently, the tumor recurred at on the posterior epipharyngeal wall after a duration of one year and three months, and was removed completely. Since then, there was no recurrence of the tumor until the present time. PMID- 2992231 TI - Increased lead levels in brain after long-term treatment with lead and dithiocarbamate or thiuram derivatives in rats. AB - Lead concentrations in blood and brain were measured in rats exposed to lead via drinking water, 0.25% Pb (12 mM), and dithiocarbamate/thiuram derivatives administered by gavage singly or in combination for 6 weeks. Sodium salts of diethyldithiocarbamate (DEDTC) and dimethyldithiocarbamate (DMDTC) were given in doses of 0.2 mmol/kg and tetramethylthiuram disulfide (thiram) and tetraethylthiuram disulfide (disulfiram) in doses of 0.1 mmol/kg twice a week. In rats that received lead plus dithiocarbamate/thiuram derivatives lead concentrations in blood and brain were significantly increased; disulfiram being most effective in increasing lead levels, followed by thiram, DMDTC and DEDTC. In blood, lead levels were increased 3-fold and in brain almost 4-fold after treatment with lead plus disulfiram compared to treatment with lead alone. When rats were given DEDTC or thiram by gavage (0.1 mmol/kg/day 5 days a week for 2 weeks) after cessation of the lead treatment, there was no increase in blood lead levels but in thiram-treated rats brain lead concentration was increased 2.7 fold. In rats treated with DEDTC intraperitoneally after cessation of lead treatment, both blood and brain concentrations of lead were increased. This study suggests that combined exposure of lead and dithiocarbamate/thiuram derivatives causes a substantial increase in brain levels of lead which are not always reflected in increases of blood lead levels. This interaction effect ought to be taken into consideration when evaluating the health effects of environmental and occupational lead exposure. PMID- 2992232 TI - The cardiac glycosides of Thevetia ovata A.DC. and Thevetia nereifolia Juss. ex Stend. PMID- 2992233 TI - Effects of pentazocine and concomitant clonidine on opioid receptors in the rat brain. AB - The changes in opioid receptors (Op-R) caused by repeated administration of pentazocine and the effect of concomitant clonidine were investigated. Binding of [3H] naloxone was markedly decreased in the absence of Na+, but was increased in the presence of Na+ in the diencephalon-mesencephalon of chronic pentazocine treated rats. No significant changes were observed in the cerebral cortex of pentazocine-treated rats. The pentazocine-induced changes in Op-R were abolished by the concurrent use of clonidine, an alpha-adrenergic agonist, which has been shown to relieve the withdrawal symptoms of morphine. This result indicated that the behavioral action of clonidine can also be observed at the Op-R level. PMID- 2992235 TI - Long-term beta 1-selective adrenergic blockade and adrenergic receptors in human subcutaneous adipocytes. AB - The influence of beta-adrenergic blockade with metoprolol, a beta 1-selective agent, on the adrenergic regulation of lipid mobilization was explored in subcutaneous adipocytes removed from 13 patients with essential hypertension. Treatment with metoprolol, which was associated with adequate beta-adrenergic blockade and an antihypertensive effect, resulted in a significant increase (p less than 0.05) in the binding of the beta-adrenergic antagonist (-)-(3H) dihydroalprenolol and a 50% increase (p less than 0.01) in the maximum lipolytic response to the beta-adrenergic agonist isopropylnoradrenaline. In 7 patients with normotriglyceridaemia the total plasma triglyceride level increased significantly (p less than 0.025) during metoprolol treatment, a change that was due to an increase in the very low density lipoprotein triglycerides. The findings suggest that chronic treatment with the beta 1-selective adrenergic blocker metoprolol leads to a significant increase in beta-adrenoceptor density and an increase in the lipolytic response to beta-adrenergic agonists. This latter finding may, in some measure, account for the increased plasma triglyceride level observed. PMID- 2992234 TI - Mucinous carcinoma of the jejunum. Report of a clinical case. PMID- 2992236 TI - Low density lipoprotein receptor activity in human leukemic cells--relation to chromosome aberrations. AB - Chromosome analysis and low density lipoprotein (LDL) receptor activity of leukemic cells from 38 patients with acute non-lymphocytic leukemia were correlated. Clonal chromosome aberrations were found in 22 patients, and an extra chromosome 8 was found in 7 of them. LDL receptor activity was significantly higher in patients with an extra chromosome 8 than in patients with other abnormalities or a normal karyotype. Chromosome 8 may harbor genes of importance for the expression of the LDL receptor. PMID- 2992237 TI - Morbid obesity, gastric plication and a severe neurological deficit. AB - A 39-year-old man had protracted vomiting after gastric plication for morbid obesity. Within three months he lost 53 kg in weight and developed neuromuscular weakness, especially in the lower extremities. Clinical and laboratory studies suggested both radicular and peripheral neuropathy. One year later the condition was only marginally improved: he took only few steps unsupported. The apparent etiology is malnutrition but the primary cause remained unknown. PMID- 2992238 TI - Infections in neurosurgery: a randomized comparison between silk and polyglycolic acid. AB - Silk or polyglycolic acid (PGA) was chosen at random for 1,011 patients operated on in the department during a 19 month period in 1981. There was no difference in the incidence of serious infections between the two groups. A significantly higher incidence of suture fistulas in the silk group indicates the use of PGA for buried sutures. PMID- 2992239 TI - Our experience with thin needle percutaneous transhepatic cholangiography. PMID- 2992241 TI - Leukotrienes: biosynthesis, metabolism, and analysis. PMID- 2992240 TI - [Progress in dentistry and maxillofacial surgery]. PMID- 2992242 TI - Pharmacology of the leukotrienes. PMID- 2992243 TI - Chemistry of the prostaglandins and leukotrienes. PMID- 2992244 TI - Synthesis and structure elucidation of leukotrienes. PMID- 2992245 TI - Prostaglandin endoperoxide and thromboxane A2 analogs. PMID- 2992246 TI - Analogs of leukotrienes and related lipoxygenase products. PMID- 2992247 TI - Structural determination of leukotrienes and lipoxins. PMID- 2992248 TI - [Acute haemorrhagic conjunctivitis caused by a subtype of adenovirus type 37]. PMID- 2992249 TI - [Intrascrotal malignant fibrous histiocytoma: a case report and review of the literature]. AB - A 75-year-old man visited our clinic on November 11, 1982 with the complaint of a painless mass in the right scrotum. The mass was hen's egg sized and hard. Surgery was performed under the diagnosis of testicular tumor. Right hemiscrotectomy with right inguinal orchiectomy was performed because of a scrotal skin invasion of the tumor. The spermatic cord, testis and epididymis were grossly normal. The tumor was located just beneath the testis, and it was considered to be scrotal tunicus in origin. The tumor was elastic and hard, and the cut surface showed a yellowish white and lobulated appearance. The histological diagnosis was malignant fibrous histiocytoma. Postoperatively, radiotherapy for a total dosage of 4,910 rads was given. The patient is alive and well without any sign of recurrence 19 months after operation. The 20 cases reported in Europe, America and Japan, including our case, were tabulated and some discussion is made. PMID- 2992251 TI - Primary liver tumor in a young man: a brief review of diagnostic strategies and treatment options. PMID- 2992250 TI - Magnetic resonance imaging of cavernous hemangioma of the liver: tissue-specific characterization. AB - Twenty-one patients with hepatic hemangioma, five with hepatic cysts, and 25 with primary or metastatic cancer involving the liver were studied by magnetic resonance imaging (MRI). Benign lesions (hemangiomas, cysts) were diagnosed noninvasively by CT, radionuclide studies, and/or sonography and confirmed by follow-up examinations more than 1 year later. Malignant lesions were confirmed by liver biopsy in every case. Identical multisection/multiecho techniques were used in all patients to obtain T1-and T2-weighted spin-echo (SE) and inversion recovery (IR) images. MRI detected more hemangiomas than any other imaging technique. Of 30 hemangiomas, 25 were spherical or ovoid with a homogeneous appearance and smooth, well defined margins. Cancer tended to have a heterogeneous appearance and poorly defined margins. On T2-weighted SE images obtained with 2000 msec TR and 60, 120, or 180 msec TE, hemangiomas had significantly greater contrast-to-noise ratios (C/N) than liver cancer (p less than 0.001). The SE 2000/120 sequence provided the single most useful image for distinguishing hemangiomas from cancers. When morphologic criteria are used in conjunction with measured C/N, MRI correctly distinguished cavernous hemangiomas from liver cancer with 90% sensitivity, 92% specificity, and an overall accuracy of 90%. Cysts had a low signal intensity on SE 500/30 images and could often be distinguished from hemangiomas and cancers that were nearly isointense relative to liver. IR images were sensitive for lesion detection but provided no tissue specific information. The data indicate that T2-weighted SE imaging may become the procedure of choice for distinguishing cavernous hemangioma from liver cancer. PMID- 2992252 TI - Computed tomography of Krukenberg tumors. AB - Computed tomography (CT) of three patients with Krukenberg tumor was reviewed retrospectively. CT showed large, lobulated, multicystic masses with soft-tissue components, indistinguishable from primary ovarian carcinoma. Indeed, CT and sonography of all three patients were initially interpreted as primary ovarian carcinoma. The ovary is a frequent site of metastases, particularly from colon carcinoma. These can be quite large, yet diagnosis is seldom made preoperatively. Much has been written about metastatic ovarian tumor, but this is the first report in the radiologic literature about their CT features. The authors emphasize the importance of recognizing the ovary as a frequent site of metastases and the proper approach to this problem. In patients with a history of colon or gastric carcinoma, the mixed cystic and solid ovarian mass on CT should be regarded as metastatic tumor until proven otherwise. A careful search for gastrointestinal tract signs or symptoms should be done in any patient with a pelvic tumor. When CT is done for evaluation of ovarian tumor, the stomach and colon should be carefully evaluated, and the ovaries routinely examined in the preoperative CT staging of gastric or colon carcinoma. PMID- 2992253 TI - Fleischner lecture. Biokinetic impacts on structure and imaging of the lung: the concept of biologically closed electric circuits. AB - The corona complex of the lung is a collection of radiographic features surrounding a pulmonary mass. Each of its 12 components may be explained by a recently described subset of biologically closed electric circuits, the vascular interstitial closed electric circuit (VICC) system. This system is activated both by normal metabolism of tissue and by local degrading processes, such as spontaneous necrosis or hemorrhage, that lead to local electrochemical polarization of a lesion in relation to surrounding noninjured tissue. Ions, cells, and water are transported electrically in the VICC system, leading to the development of the corona complex. The VICC system is conceived to exist with the walls of arteries and veins functioning as insulators around the electrically conducting medium of blood, the plasma. Blood vessels therefore connect electrically the injured and noninjured tissues. At the capillary level, electric junctions connect plasma and interstitial fluid, which functions as an electrical conductor comparable to blood plasma. The interstitial fluid therefore completes the circuit. The VICC system can be regarded as an additional circulatory system for selective electrogenic transports, coupled directly to the mechanical circulation of blood and lymph. The injury potential represents an important energetic factor in the activation of the VICC system. It is a slowly fluctuating, attenuating, electrochemical potential inducing ebb and flow of time dependent anionic and cationic transports. The corona structures are special effects of the healing of injured tissue. The 12 radiologic signs of the corona complex have each been produced experimentally in vitro in animals and in vivo in humans during electrochemical treatment of cancers. The "A" zone is characterized radiographically by radiolucency around an electrically polarizing focal lesion. Peripheral to the A zone, a "B" zone is seen as a radiopaque region. The A and B zones are predominantly the result of an electroosmotic outflow of water from a lesion during its electropositive phase. At the interface between the A and B zones, small arches sometimes form an arcade. This configuration develops when the polarizing lesion has small protrusions at its surface. As a result of electrical edge enhancement, various elements of the interstitial tissue are transformed into radiating fibrous structures. They grow out at right angles to the surface of the lesion and serve as supporting columns for the arches. When necrotic material from a tumor is evacuated through a bronchus, ensuing collapse of the tumor will displace those radiating structures already produced.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2992254 TI - Relation between mammographic findings and hormonal receptor content in breast cancer. AB - A comparative investigation of 85 patients with unilateral primary breast cancer was performed to elucidate a possible correlation between the mammographic appearance of the cancer and the content of hormonal receptors (estrogen and progesterone). The mammographic appearance was divided into five types: (1) spiculated; (2) structural changes in the parenchyma (density); (3) calcifications; (4) circumscribed; and (5) not visible on mammogram. The hormonal content was found to be high in types 1 and 2, low in 3 and 4, and intermediate in 5. Statistically significant difference in the estrogen receptor content was found between 1 and 3; 1 and 3 + 4; 1 and 3 + 4 + 5. In progesterone receptor content, a statistically significant difference was found between 1 and 3 + 4. The results concerning estrogen receptor content within each cancer type, except type 2, are comparable to those of a Swedish study. No correlation was found between Wolfe's parenchymal patterns and hormonal receptor content, which contradicts a recent English study. Reasons for this discrepancy are discussed. PMID- 2992255 TI - Malignant thoracopulmonary small-cell ("Askin") tumor. AB - The clinical, radiographic, and pathologic features of 10 patients with documented malignant small-cell tumor of the thoracopulmonary region (Askin tumor) were reviewed. The tumor represents a distinct pathologic entity of neuroectodermal origin. Clinically, it presents as a chest-wall mass with or without pain. Its radiographic appearance is that of a soft-tissue mass with or without pleural or rib involvement, often with metastatic disease--to the skeletal system, bone marrow, thorax, and sympathetic chain. Two patients developed metastases to the adrenal gland and liver, one after autologous bone marrow transplantation. The radiologist should be aware of this entity and its pattern of metastatic spread since metastases are treated aggressively. PMID- 2992256 TI - Magnetic resonance imaging of the pelvis: evaluation of ovarian masses at 0.15 T. AB - Seventeen patients with suspected ovarian masses were evaluated by magnetic resonance imaging (MRI). MRI findings were confirmed by surgery (13 patients) or sonography and clinical follow-up (four). The study evaluated MRI characteristics of ovarian lesions using recently developed multislice and multiecho pulse techniques for a 0.15-T system. T1 and T2 relaxation times were calculated in 12 patients and although a range of values was obtained in several disease categories, diagnostic accuracy was frequently improved. MRI appearances tended to vary considerably with different pulse sequences and were particularly complex in patients with endometriosis and cystic ovarian tumors. Shortest calculated T1 and T2 values were found in hemorrhagic cysts in patients with endometriosis. Benign tumors with thick fibrous pseudocapsules had longer T1 values. Inflammatory masses and malignant ovarian tumors had significantly longer T1 and T2 values and relaxation times in a patient with mucinous cystadenoma varied within the complex mass. PMID- 2992257 TI - Dynamic study of supratentorial gliomas with L-methyl-11C-methionine and positron emission tomography. AB - The regional kinetics of intravenously injected L-methyl-11C-methionine (11C-L methionine) in the brain was investigated by positron emission tomography (PET) in 14 patients with gliomas. In both tumor and unaffected brain the tracer uptake reached a nearly constant level in 5 min or less. The ratio between the uptake of 11C-L-methionine by high-grade tumors and the uptake by unaffected brain was 1.9 4.8. In two cases of low-grade astrocytoma the ratio was 0.8-1.0. High uptakes of 11C-L-methionine occurred in gliomas even in the absence of blood-brain barrier defects as observed by other methods. This indicates that besides active transport of amino acid, a larger extracellular space in tumor as compared with unaffected brain tissue may also contribute to the increased uptake of 11C-L methionine--derived radioactivity. In some patients delineation of the tumors was improved by use of PET with 11C-L-methionine as compared with computed tomography, angiography, and, in some instances, PET with 68Ga-EDTA. PET with 11C L-methionine permits better evaluation of the tumor extent and may affect preoperative grading. PMID- 2992258 TI - Nutritional pharmacology. PMID- 2992259 TI - Technetium-99m-pyrophosphate scintigraphy in patients with suspected acute myocardial infarction: impact of interobserver variability. AB - Technetium-99m-pyrophosphate (TcPYP) scintigraphy may have great value in patients with suspected acute myocardial infarction (AMI), but interobserver variability undoubtedly has adverse impact on predictive value. TcPYP scintigrams for 133 (80%) of 166 consecutive patients admitted for suspected AMI were interpreted independently by three experienced readers. Although there was complete agreement for 87 interpretations (65%), major discrepancies (i.e., at least one positive and one negative reading on the same scan) occurred for 28 scans (21%). To assess predictive accuracy, patients were categorized as follows: 36 had definite AMI manifest by new ECG Q waves and/or CK-MB evidence of AMI (group I), 56 were classified as possible AMI (group II), and 41 had AMI excluded (group III). Using only the definitive diagnostic categories (groups I and III), accuracy for each reader approximated 0.68, with no single reader being correct more often than any other. PMID- 2992261 TI - A variable feed rate mechanism for fluidized bed asbestos generators. AB - A simple and inexpensive dust feed mechanism has been designed for use with a two phase fluidized bed generator (FBG). The mechanism is especially useful for generating asbestos aerosols, but may be used with other dusts as well. Using this system, a steady state concentration (39.1 fibers/cc greater than 5 microns in length +/- 6.2%) of asbestos aerosol was maintained in an inhalation chamber for five hours. In addition, FBG output concentration was easily adjusted and quickly equilibrated (within 10 minutes). The system provides a good technique for generating asbestos aerosols for day-long animal exposures. PMID- 2992260 TI - Effects of ryanodine on cat papillary muscle and isolated rat heart. AB - The neutral alkaloid, ryanodine, has several actions on cardiac muscle. To delineate better its mode of action, we studied ryanodine's effect upon contracting cat papillary muscles under changing loading conditions and stimulation frequencies. We also studied ryanodine's physiologic and metabolic effects upon isolated rat hearts. The results of our study suggest the following: (1) ryanodine causes both decreased release and decreased uptake of calcium by the sarcoplasmic reticulum; (2) elevation of high-energy phosphates secondary to decreased energy requirements is due to decreased calcium availability to the myofilaments during systole; (3) the slowed or incomplete relaxation caused by ryanodine may be a stimulus for myosin phosphorylation; (4) ryanodine probably decreases calcium movement through the sarcolemma and so increases adenosine and inorganic phosphate and decreased cyclic adenosine monophosphate (AMP) concentration in the myocardium; and (5) the effect of ryanodine on altered loading conditions and contraction velocities can be understood in terms of decreased calcium availability to the myofilaments. PMID- 2992262 TI - A critique of MSHA procedures for determination of permissible respirable coal mine dust containing free silica. AB - The literature in selected technical subject areas related to the Mine Safety and Health Administration's (MSHA) respirable mine dust control policy was critically reviewed. Specific topics included: (1) the technical aspects of respirable dust sampling, (2) the development of a sampling strategy and (3) currently used analytical techniques for free silica content of respirable coal mine dust. The 2.0 Lpm MSHA respirable dust sampling flow rate does not conform with published results which indicate that to simulate existing pulmonary particle deposition models, the sampling flow rate should range from 1.4 to 1.7 Lpm. MSHA's sampling strategy focuses on controlling respirable coal mine dust with both area and personal samples of workers in selected occupations or activities. Many uncertainties are encountered as area samples are used to estimate personal exposures. Although all of the analytical methods for crystalline free silica are sufficiently sensitive to be able to detect and quantitate free silica at environmentally significant concentrations, they are all plagued with similar difficulties. Analytical standards representing the various forms of silica are not available. Available analytical methods do not differentiate among polymorphs of silica. Recommendations are presented to resolve identified problem areas. PMID- 2992263 TI - Serum fatty acids, apolipoproteins, selenium and vitamin antioxidants and the risk of death from coronary artery disease. AB - The independent association of serum concentrations of saturated and polyunsaturated fatty acids, apolipoproteins AI and B, selenium and vitamins A and E with the risk of death from coronary artery disease (CAD) was studied in 92 persons with no previous myocardial infarction, who died from CAD during a 5-year follow-up, and their 92 1-to-1 matched controls. Case-control pairs came from a randomly drawn population sample of approximately 12,000 persons aged 30 to 64 years from 2 provinces of eastern Finland, an area with exceptionally high CAD mortality. Control subjects were matched for sex, age, serum cholesterol, mean arterial pressure, tobacco consumption and history of cardiovascular diseases. The persons who died of CAD had lower serum esterified arachidonic acid concentrations before follow-up than the control subjects (41 vs 48 mg/liter, p = 0.05), and this difference was greater for pairs with no chest pain on effort (36 vs 50 mg/liter, p less than 0.05). The adjusted risk of CAD death in persons with a serum polyunsaturated to saturated (P/S) fatty acid ratio of 0.28 or less (in the lowest tertile) was 3.5-fold (95% confidence interval [CI], 1.5 to 8.2) compared with those with higher serum P/s ratios in a multivariate logistic model and 5.6-fold (95% CI 1.6 to 19.8) for pairs with no chest pain on effort. A low serum apolipoprotein AI concentration (1.25 g/liter or less, in the lowest tertile) was associated with a 2.5-fold (95% CI 1.1 to 5.7) adjusted risk of CAD death among the chest pain-free persons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992264 TI - The effects of the fiber components pectin, cellulose and lignin on serum cholesterol levels. AB - Eight-week randomized cross-over studies in three separate groups of 10 healthy volunteers were undertaken to determine the effects of daily dietary supplementation with pectin (12 g/day), cellulose (15 g/day) and lignin (12 g/day) on serum lipid levels. Detailed dietary records were kept throughout the study and there was no significant change in dietary intakes except for the fiber supplement. Neither pectin, cellulose, nor lignin significantly altered serum total cholesterol, triglycerides, high-density lipoprotein cholesterol, or the ratio of high-density lipoprotein to total cholesterol in healthy normolipidemic subjects over four weeks. PMID- 2992265 TI - Ultrastructure of cystosarcoma phyllodes and fibroadenoma. A comparative study. AB - Electron microscopic studies were conducted on 11 cases of cystosarcoma phyllodes (6 benign, 3 malignant, 2 recurrent), 1 stromal sarcoma, and 15 cases of fibroadenomas. Particular emphasis was placed on examination of the epithelial stromal junction. The epithelial cells were basically similar in all cases. The basal lamina of fibroadenomas tended to be reduplicated, whereas in cystosarcoma phyllodes the basal lamina was less reduplicated and even focally absent. A continuous layer of delimiting fibroblasts was not present in either type of tumor. Stromal cells were generally fibroblasts, with 2 of 15 being fibroadenomas and 2 of 11 cystosarcoma phyllodes showing myofibroblastic differentiation. The stromal sarcoma also showed smooth muscle differentiation. The authors conclude that the epithelial-stromal junction is abnormal in both fibroadenomas and cystosarcomas, but the abnormality is different in these two tumors. Increasing prominence of stromal cell junctions and of lysosomes also correlated with the diagnosis of cystosarcoma, particularly the malignant variant. PMID- 2992266 TI - Hematopoietic cell surface markers on metastatic small cell carcinoma detected with monoclonal antibodies. AB - Using a panel of monoclonal antibodies, cells from lymph node biopsies have been examined in three patients with small cell carcinoma presenting with cervical lymphadenopathy. Two patients had small (oat) cell carcinoma of the lung; in the third patient, a primary tumor was not found. Two lymph node biopsies showed typical small (oat) cell carcinoma, and one was an intermediate cell variant; in the last, lung biopsy showed small (oat) cell carcinoma. Electron microscopy demonstrated desmosomes in all three tumors. In each case, lymph node cell suspensions were examined by indirect immunofluorescence with the use of a panel of monoclonal antibodies to antigens usually associated with lymphoid or myeloid cells. In two of the three cases malignant cells were positive with the lymphoid marker BA-2; in two cases malignant cells were positive with OK1a1, a marker for the Ia-like antigen (HLA-DR); and in one case malignant cells were positive with My-1. Caution is needed in the interpretation of cell surface marker studies in the differential diagnosis of small round cell tumors. PMID- 2992267 TI - An outbreak of Norwalk-related gastroenteritis at a boys' camp. AB - An acute gastrointestinal tract illness affected 213 (52%) of 407 campers and 64 (53%) of 121 staff members attending a boys' camp in the Catoctin Mountains of Maryland during the summer of 1981. Nausea was the predominant symptom for ill campers and staff members (73%), but more staff members experienced diarrhea (49%) than did campers (9%). Twenty-three individuals had more than one episode of illness compatible with the case definition. Eight of nine paired blood specimens from ill staff members showed a fourfold increase in antibody titer to Norwalk virus by radioimmunoassay. Environmental inspections and laboratory tests failed to implicate a common source of exposure. PMID- 2992268 TI - Hepatocellular carcinoma simulating amebic liver abscess: report of a case and analysis of current diagnostic methods. AB - Radiological and serological tests are valuable tools commonly used in diagnosing amebic liver abscess. The limitations in sensitivity and specificity of the various imaging procedures should be recognized in order to avoid misinterpretation of test results. Serodiagnostic methods present potential pitfalls primarily because of their frequent inability to differentiate previous disease from active infection, and must therefore be interpreted carefully. PMID- 2992269 TI - Association between urinary zinc excretion and lymphocyte dysfunction in patients with lung cancer. AB - Patients with bronchogenic carcinoma often have low serum zinc concentrations and sometimes have markedly elevated renal zinc losses. Since normal zinc metabolism is critical for the proper function of T lymphocytes and natural killer cells, the effect of zinc status on T cell phytohemagglutinin response and peripheral blood lymphocyte natural killer cell activity was studied in patients with lung cancer. Mean (+/- SEM) serum zinc concentration in 75 patients with cancer was 67.4 +/- 2.2 micrograms/dl versus 96.0 +/- 8.0 micrograms/dl for normal subjects. Patients with low serum zinc levels (less than 70 micrograms/dl) had significantly higher urine zinc excretion than patients with normal serum zinc levels (1,385 +/- 240 micrograms per 24 hours versus 392 +/- 107 micrograms per 24 hours) (p less than 0.001). This pattern of zinc concentrations (i.e., low serum zinc in combination with high urine zinc) is typical of patients with mild zinc deficiency, and suggests that a mild chronic zinc deficiency state was present in some of these patients. When lymphocyte data were analyzed according to serum zinc concentrations and urinary zinc excretion, low serum zinc concentration and high urine zinc excretion both correlated with depressed T cell phytohemagglutinin response (p less than 0.005 and p less than 0.001, respectively). For instance, mean maximal phytohemagglutinin response in patients with urinary zinc excretion of more than 700 micrograms per 24 hours was 22,132 +/- 3,201 cpm (n = 14) compared with 68,130 +/- 6,850 cpm for patients with normal zinc excretion (n = 7). Peripheral blood lymphocyte natural killer cell activity did not correlate with either serum or urine zinc values. Oral zinc sulfate (220 mg, three times daily for six weeks) was then administered to patients with hyperzincuria (mean = 992 micrograms per 24 hours). Zinc supplemented patients had normalization of T cell phytohemagglutinin response after zinc therapy, whereas control patients demonstrated continued T cell dysfunction. Natural killer cell activity did not change in either group during the study period. These data suggest that a mild subclinical zinc deficiency state may exist in some patients with lung cancer and may be an important cause of abnormal T cell function. Furthermore, zinc supplementation may be useful to improve lymphocyte function in selected patients. Whether zinc supplementation would alter the course of the disease or the patient's prognosis is presently unknown. PMID- 2992270 TI - DNA polymorphism and the susceptibility to diabetes. AB - We have studied the relationship between the DNA polymorphisms and the development of diabetes in Caucasians and US black populations. The analyses suggest that disease susceptibility differs in the two populations. In a large population with low frequency of diabetes, DNA fragments detected in the studies of the human insulin gene and HLA-DR regions seem to have a minor role in the development of diabetes. However, the currently available data also show that alleles with insertion in the 5'-flanking region of the insulin gene in the US black populations induce noninsulin-dependent diabetes fivefold more often than in white populations. To draw a more reliable conclusion, further DNA polymorphism surveys in different ethnic groups need to be conducted. PMID- 2992272 TI - Familial glioblastoma multiforme without neurofibromatosis. AB - Glioblastoma multiforme is a rapidly growing anaplastic primary tumor of the central nervous system. It is frequently associated with neurofibromatosis or other hamartomatous syndromes, and in such kindreds may be found in several generations. We describe a highly inbred family in which glioblastoma multiforme presents in males only as an isolated central nervous system neoplasm on the right side, without evidence of other underlying genetic disease. It is speculated that in this family the tumor develops as the consequence of an autosomal recessive or an X-linked recessive mutation. In addition, the gene for cystic fibrosis segregates in this family and an undefined autosomal recessive malformation syndrome was detected. PMID- 2992271 TI - Antifolate-induced misincorporation of deoxyuridine monophosphate into DNA by cells from patients with the fragile X syndrome. AB - The fragile site at Xq27 is expressed in vitro under conditions that lead to decreased intracellular thymidine triphosphate concentration, a condition which has also been shown to promote the misincorporation into DNA of deoxyuridine monophosphate (dUMP) in place of thymidine. We tested for increased whole-cell misincorporation of dUMP as a possible molecular mechanism for the expression of the fragile X abnormality. Neither deoxyuridine triphosphatase nor uracil-DNA glycosylase, the two enzymes that normally prevent the accumulation of dUMP in DNA, was deficient in fragile X syndrome cells. Misincorporation of dUMP occurred in comparably low levels in both normal and fragile X syndrome lymphoblasts. Although these results provide strong evidence against generalized misincorporation of dUMP in fragile X syndrome cells, a substantial real difference present at Xq27 might not be detected in these studies of whole cells containing the diploid chromosome complement. PMID- 2992273 TI - Human myometrial adrenergic receptors during pregnancy: identification of the alpha-adrenergic receptor by [3H] dihydroergocryptine binding. AB - The radioactive alpha-adrenergic antagonist [3H] dihydroergocryptine binds to particulate preparations of term pregnant human myometrium in a manner compatible with binding to the alpha-adrenergic receptor (alpha-receptor). [3H] Dihydroergocryptine binds with high affinity (KD = 2 nmol/L and low capacity (receptor concentration = 100 fmol/mg of protein). Adrenergic agonists compete for [3H] dihydroergocryptine binding sites stereo-selectively ([-]-norepinephrine is 100 times as potent as [+]-norepinephrine) and in a manner compatible with alpha-adrenergic potencies (epinephrine approximately equal to norepinephrine much greater than isoproterenol). Studies in which prazosin, an alpha 1 antagonist, and yohimbine, and alpha 2-antagonist, competed for [3H] dihydroergocryptine binding sites in human myometrium indicated that approximately 70% are alpha 2-receptors and that 30% are alpha 1-receptors. [3H] dihydroergocryptine binding to human myometrial membrane particulate provides an important tool with which to study the molecular mechanisms of uterine alpha adrenergic response. PMID- 2992274 TI - Reproductive patterns and the risk of gestational trophoblastic disease. AB - The relation between reproductive pattern and the risk of gestational trophoblastic disease was evaluated in a case-control study conducted in Northern Italy on 310 women with histologically confirmed gestational trophoblastic disease and two control groups consisting of 290 obstetric subjects and 394 patients in hospital for acute, nonobstetric, nongynecologic conditions. Compared to that for nulliparous women, the estimated age-adjusted relative risk of trophoblastic disease for parous women was 0.6 (90% confidence limit = 0.4 to 0.9) when obstetric controls were used as a comparison group and 0.4 (95% confidence limit = 0.2 to 0.6) compared with other controls. Conversely, a history of spontaneous abortions was associated with elevated risk of gestational trophoblastic disease, and the risk increased significantly with increasing number of spontaneous abortions. When the combined effect of parity and spontaneous abortions was considered, the major factor influencing the risk of gestational trophoblastic disease was the existence of one or more previous term pregnancies. PMID- 2992275 TI - Congenital herpes simplex type II infection. AB - A case of congenital herpes simplex virus type II infection acquired in utero is reported. The issue of congenital infection due to the herpes simplex viruses is discussed. PMID- 2992277 TI - Surgical problems in the management of giant fibroadenoma of the breast. AB - Giant intracanalicular fibroadenoma is reported to be the most common cause of a massive, deforming enlargement of one breast in the female adolescent. It occurs infrequently and has a varied histopathologic pattern. It is suggested that obstetricians and gynecologists should have an awareness of this potentially malignant tumor of the breast with its associated problems of diagnosis and surgical management. Consideration of these huge breast tumors in relation to other similar breast lesions is presented in order to emphasize differential characteristics and treatment. PMID- 2992276 TI - Intraperitoneal immunotherapy of epithelial ovarian carcinoma with Corynebacterium parvum. AB - Corynebacterium parvum was administered intraperitoneally to 21 patients with epithelial ovarian cancer. Nineteen patients had surgically measurable disease and two received adjuvant therapy. Surgically confirmed responses were documented in six of 19 patients (31.6%), with two complete responses (10.5%) and four partial responses (21.1%). Three patients (15.8%) had stable disease, and 10 patients (52.6%) had disease progression. The mean survival of the patients who had a complete response was 35.5 months; the four patients who had a partial response the mean survival was 26.6 months, and of the nonresponders the mean survival was 12.6 months (p less than 0.02). The mean survival of the entire group was 18.2 months. Initial response and patient survival correlated with the amount of disease pretreatment. Thus six responding patients had less than or equal to 5 mm maximum diameter tumors, that is, minimal residual disease. Toxicity in the 86 courses of therapy included abdominal pain in 78% of cases, fever in 56%, nausea in 40%, and vomiting in 22%. Stimulation of cytotoxic lymphocytes resulted from the administration of C. parvum, which induced a significant increase of both intraperitoneal natural killer lymphocyte cytotoxicity and antibody-dependent cell-mediated cytotoxicity in six of nine patients tested; these two types of cytotoxicity correlated with response to therapy and may be partially responsible for the surgically documented tumor regression. While the clinical usefulness of intraperitoneal C. parvum is limited because of its toxicity, intraperitoneal immunotherapy may prove useful in patients with minimal residual ovarian cancer when more refined agents become available. PMID- 2992278 TI - A double-blind comparative study of Estraderm and Premarin in the amelioration of postmenopausal symptoms. AB - Patients whose postmenopausal symptoms were being satisfactorily controlled with conjugated equine estrogens, either 0.625 mg/day (n = 57) or 1.25 mg/day (n = 67), participated in a study that compared the efficacy of these oral regimens with that of 17 beta-estradiol, 0.1 mg/day, administered through intact skin. The study was a multicenter, double-blind, randomized, parallel-group trial during which two thirds of the patients who received each dosage of conjugated equine estrogens were changed to an estradiol transdermal system while the remainder continued with conjugated equine estrogens. A total of 124 patients was included in the analysis of efficacy. The analysis revealed no significant differences between the estradiol transdermal system and conjugated equine estrogens in control of hot flushes or other postmenopausal symptoms and no statistically significant differences between treatment groups in regard to estrogen-related side effects. Minor topical reactions to the transdermal systems were reported during only about 20% of study weeks. Thus, transdermal estradiol, 0.1 mg/day, appears to be equally effective as conjugated equine estrogens, 0.625 or 1.25 mg/day, for controlling postmenopausal symptoms and is well tolerated. PMID- 2992279 TI - Pharmacokinetics and pharmacodynamics of transdermal dosage forms of 17 beta estradiol: comparison with conventional oral estrogens used for hormone replacement. AB - This open-label, multiple-crossover study compared the pharmacokinetics and pharmacodynamics of transdermal 17 beta-estradiol and two oral forms of estrogen replacement therapy in postmenopausal women. The transdermal systems delivered either 0.025, 0.05, or 0.1 mg/day; oral dosages were 2 mg of micronized 17 beta estradiol or 1.25 mg of conjugated equine estrogens. Transdermal estradiol provided serum and urinary levels of estradiol conjugates typical of the early follicular phase of the premenopausal woman and an estradiol/estrone ratio that approximated 1. The increments of both serum and urinary estradiol showed dose proportionality. Serum levels of estradiol obtained 24 hours after oral administration of estrogens were in a range similar to the steady-state levels obtained with transdermal estradiol delivery. Oral estrogens, however, induced an excessive rise in estrone to levels far beyond those observed in premenopausal women. Continuous application of transdermal estradiol over 3 weeks did not result in any accumulation of estradiol or estradiol conjugates. After only three doses of oral estrogens, there were signs of retention of estrogens. Suppression of gonadotropins by oral and transdermal administration of estrogens was in a similar range. This observation supports the conclusions that levels of circulating estradiol are relevant to efficacy, and that excessively high levels of estrone after oral administration of estrogens merely represents a nonphysiologic precursor or metabolite pattern. PMID- 2992280 TI - A prospective study of the ophthalmologic findings in the acquired immune deficiency syndrome in Africa. AB - A prospective study of ophthalmologic findings in 20 African patients with the acquired immunodeficiency syndrome (Group 1) and nine patients with acquired immune deficiency syndrome related complex (Group 2) disclosed that 11 patients in Group 1 and two patients in Group 2 had ocular involvement. Cotton-wool spots, Roth spots, and hemorrhages were frequent features in Group 1. In this series, unlike most others, perivasculitis occurred in five patients whereas cytomegalovirus retinitis and ocular involvement of Kaposi's sarcoma were not observed. These African patients also differed from those previously described in the high proportion of women (11 of 29) and the lack of the usual risk factors (homosexuality, intravenous drug abuse, and hemophilia). PMID- 2992281 TI - Progressive herpetic corneal endotheliitis. PMID- 2992282 TI - Malignant fibrous histiocytoma in rats at sites of implanted millipore filters. AB - Soft-tissue tumors developing in rats at the site of implanted Millipore filters showed the structural, cytochemical, and biologic characteristics of the malignant fibrous histiocytomas occurring in man. Evidence from this tumor model suggests that malignant fibrous histiocytomas can arise from pluripotential mesenchymal stem cells which are not derived from the mononuclear/phagocytic system and that chronic inflammation and scarring may predispose to their development. PMID- 2992284 TI - Diurnal-stimulated and stress-induced ACTH release in rats is mediated by ventral noradrenergic bundle. AB - Female rats were bilaterally injected with 3 micrograms of 6-hydroxydopamine (6 OHDA) dissolved in 0.2 microliter saline, via a glass micropipet stereotaxically implanted into the ventral noradrenergic-ascending bundle (VNAB). This bundle conveys most of the catecholaminergic innervation to the paraventricular nuclei and originates from the locus coeruleus and from two medullary groups of neurons (A1 and A2). Two weeks after injection, and 1 wk after the subsequent implantation of an arterial cannula, serial blood samples were taken from each rat over a 36-h period for assay of basal secretion patterns of ACTH and corticosterone (C) by radioimmunoassay and radiocompetition, respectively. Other blood samples were collected at short intervals over a 2-h period to explore the stress-ether responses of both hormones. Effects of 6-OHDA injections on catecholaminergic innervation were attested by the striking decrease in the histofluorescence of hypothalamic catecholamines and by the 86% drop in the hypothalamic noradrenaline concentrations measured by high-performance liquid chromatography at constant dopamine titers. Compared with control, sham-lesioned rats, pharmacological destruction of the VNAB by 6-OHDA led to 1) obliteration of the circadian patterns for ACTH and C and the emergence in their place of ultradian fluctuations of reduced amplitude above base-line levels and 2) 80% inhibition of the ACTH stress response which correlated with a short-lived, depressed C response. These results are discussed within the framework of the controversial literature on the mechanisms by which catecholamines may control corticotropic function. PMID- 2992283 TI - Ascorbate in aqueous humor protects against myeloperoxidase-induced oxidation. AB - Chemotactic factors can cause polymorphonuclear leukocytes to release the contents of azurophilic granules, including the enzymes beta-glucuronidase and myeloperoxidase. In the presence of aqueous humor from the anterior chamber of the rabbit eye, the supernatant from stimulated leukocytes contains beta glucuronidase, but myeloperoxidase is not detectable. Studies with aqueous humor and partially purified human myeloperoxidase suggest that this phenomenon is not due to a failure of enzyme release. The factor responsible for the inability to detect MPO in the assay system is heat-labile, dialyzable, and reversed by ascorbate oxidase. Comparable assay inhibition is produced by ascorbic acid at a concentration present in either human or rabbit aqueous humor. The ability of aqueous humor to protect against myeloperoxidase-induced oxidation may contribute to several diverse phenomena, including the susceptibility of the eye to Candida infection and a prolonged half-life for several inflammatory mediators in the anterior chamber. PMID- 2992285 TI - Role of serotonin type 2 receptors in regulation of aldosterone production. AB - It is well recognized that serotonin stimulates aldosterone production by the adrenal glands. To investigate the possible roles of serotonin type 1 and 2 receptors in the regulation of aldosterone production, we examined the effects of cyproheptadine (a serotonin antagonist that inhibits both type 1 and 2 receptors) and ketanserin (a serotonin type 2 selective antagonist) on aldosterone and cAMP production in collagenase dispersed rat adrenal capsular cells. Serotonin, ranging in concentration from 10(-9) to 10(-3) M, significantly increased aldosterone production in a dose-dependent fashion after 2 h of incubation at 37 degrees C. Cyproheptadine and ketanserin showed comparable inhibitory effects on basal aldosterone production. These serotonin antagonists preferentially inhibited serotonin-induced aldosterone production. Serotonin significantly increased cAMP production at a dose of 10(-6) M. Both cyproheptadine and ketanserin significantly decreased basal cAMP production at doses of 10(-5) M. These serotonin antagonists preferentially inhibited serotonin-stimulated cAMP production. These results suggest that adrenal serotonin type 2 receptors may be coupled with adenylate cyclase activity and that these receptors are involved in the regulation of aldosterone production. Whether serotonin plays an important role in the regulation of aldosterone secretion in vivo remains to be elucidated. PMID- 2992286 TI - Is intestinal peptide transport energized by a proton gradient? AB - Transport of intact peptides, followed by intracellular hydrolysis in the intestinal mucosal cells, plays an important role in the absorption of protein digestion products in the mammalian small intestine. Even though earlier studies on peptide absorption in intact-tissue preparations have indicated that peptides are transported by an active Na+-dependent mechanism, recent studies with purified brush-border membrane vesicles have unequivocally demonstrated that Na+ does not play a direct role in the translocation of peptides across the membrane. Like most amino acids, peptides are also transported as zwitterions. However, peptide transport causes depolarization of the brushborder membrane in intact mucosal cells as well as in purified membrane vesicles, and the depolarization is the result of a net transfer of positive charge across the membrane during peptide transport. This electrogenic nature of peptide transport is observed even in the absence of Na+. Peptide transport is enhanced by an interior-negative membrane potential and inhibited by an interior-positive membrane potential. An inward proton gradient stimulates peptide transport, and this stimulation is reduced when the proton gradient is subjected to rapid dissipation by the presence of a proton ionophore. These observations strongly suggest that peptides are cotransported with protons in the intestine. There is substantial evidence for the existence of an inward proton gradient in the mammalian small intestine, and therefore it is very likely that this proton gradient is the in vivo energy source for the uphill transport of peptides. The Na+-H+ exchanger in the brush border membrane, in conjunction with Na+-K+-ATPase at the basolateral membrane, is probably responsible for the generation and maintenance of the proton gradient and may thus be involved indirectly in the intestinal absorption of peptides. PMID- 2992287 TI - Effect of dietary cellulose on site of lipid absorption. AB - The effect of dietary cellulose on the localization within the small intestine of isotopically labeled triglyceride (TG) and cholesterol (CH) from a test meal was investigated. Feeding a 20% cellulose meal resulted in greater quantities of 14C TG present in both the contents and mucosa of the distal intestine compared with a fiber-free control meal. In contrast, cellulose had no effect on the localization of CH within either the intestinal contents or the mucosa. Accumulation of TG within the intestine was not due to differences in stomach emptying, as the emptying rate was similar for both TG and CH. Within the bulk phase TG must be hydrolyzed by pancreatic lipase before it is available for cellular uptake at the microvillus membrane, whereas CH requires no hydrolysis. The greater amount of TG, but not of CH, within the intestine suggests that cellulose can interfere with lipase activity in vivo. Consequently, cellulose can delay TG hydrolysis and increase the amount of lipid absorbed in the ileum. PMID- 2992288 TI - Uptake and release of gamma-aminobutyric acid in guinea pig gallbladder. AB - The presence of gamma-aminobutyric acid (GABA)-ergic neuron in guinea pig gallbladder was investigated by measuring GABA contents and glutamate decarboxylase (GAD) activity and by demonstrating the uptake and release of [3H]GABA. GABA and GAD are both present in the gallbladder, and a positive correlation in regional distribution was observed among GABA, GAD, and the number of ganglion cells. The uptake of [3H]GABA by the gallbladder showed two saturable components; a high-affinity component (Km = 23.3 microM, Vmax = 7.63 nmol X g-1 X 10 min-1) and a low-affinity component (Km = 515 microM, Vmax = 57.1 nmol X g-1 X 10 min-1). These high-affinity and low-affinity transport systems corresponded to those obtained in the presence of beta-alanine and L-2,4-diaminobutyric acid, respectively, thereby suggesting the presence of neuronal and nonneuronal GABA transport systems in this tissue. Electrical transmural stimulation produced an increase in [3H]-GABA release from the isolated gallbladder preloaded with [3H]GABA, in the presence of beta-alanine. The stimulation-evoked release of [3H]GABA was prevented by calcium-free medium containing 1 mM EGTA and tetrodotoxin, thereby indicating that the released GABA originates from the nerve terminals. These results provide evidence for the presence of GABA-ergic neurons in the guinea pig gallbladder. PMID- 2992289 TI - Electrogenicity of the Na-K-ATPase pump in bullfrog cornea epithelium. AB - The effect of changing the K concentration in the aqueous solution was studied in the frog cornea. In general, when the K concentration was increased from 4 to 20 or 79 mM, the transepithelial PD and resistance decreased. If K was decreased from 79 to 4, 20 to 4, or 4 to 0 mM, or any other combination, the PD and resistance increased. These are normal PD responses. If after a K-free period of more than 5-10 min the K was increased to 4 mM, the PD increased, an anomalous PD response. If K was increased from 0 to 20 mM, there was an initial PD increase (anomalous response) followed by a PD decrease (normal response). If K was increased from 0 to 79 mM the PD decreased, normal response only. The resistance decreased with every increase in K concentration. Anomalous responses were abolished in Na-free solutions and in the presence of both 10(-3) M ouabain and 10(-4) M vanadate in the aqueous solution. We interpret the results on the basis of two pathways, a simple K-conductive pathway and an electrogenic Na-K-ATPase pump pathway with more Na's than K's per cycle. The normal or anomalous PD responses to changes in aqueous K concentration depend on the relative resistance of the two pathways. PMID- 2992291 TI - Developmental changes in the phosphaturic response to parathyroid hormone in the rat. AB - The need for young, immature rats to maintain positive phosphate balance for growth is well recognized. However, whether this process is associated with a resistance to the phosphaturic effect of parathyroid hormone (PTH) is not clear. In these experiments we examined the effect of PTH on urinary phosphate and cAMP excretion in rats at 3, 6, 12, and 20 wk of age. Clearance experiments were performed in acutely thyroparathyroidectomized (TPTX) rats fed a normal phosphate diet (0.86%). Basal fractional excretion of phosphate (FEPi) was low in all TPTX rats (less than 1%). The phosphaturic response to a high dose of PTH (1 U X kg-1 X min-1) increased with development (from 4 to 29%). The responses to increasing doses of PTH demonstrated a decrease in sensitivity to PTH in 6- compared with 20 wk-old rats. Urinary cAMP excretion (either per milliliter glomerular filtrate or per gram kidney weight) following PTH was not different among 6-, 12-, and 20-wk old rats, thus demonstrating a dissociation between the increase in phosphate excretion and cAMP excretion. These results indicate that the phosphaturic response to PTH is blunted in immature, acutely TPTX rats and that the phosphaturia increases progressively with development. PMID- 2992290 TI - Role of myosin light-chain phosphorylation and microtubules in stress fiber morphology in cultured mesangial cells. AB - Induced elevations in intracellular cAMP caused remarkable shape changes in cultured mesangial cells that were prevented by prostaglandin E2 (PGE2). The purpose of this study was to determine the morphological and physiological basis for these alterations. Coincident with elevated levels of cAMP and shape change there was rapid dissolution of stress fibers. Cytochalasin B treatment also caused shape change and stress fiber dissolution. Stress fibers were visualized by fluorescence microscopy after appropriate staining. In such cells, microtubules were intact. PGE2 prevented stress fiber loss as well as shape change if it was included in the medium from the outset, or restored stress fibers and normal morphology within 30 min if it was added following shape change. Agents that depolymerize microtubules also prevented both stress fiber loss and shape change. Coincident with stress fiber loss and shape change induced by elevations of cAMP, there was a decrease in the rate of [32P]-orthophosphate incorporation into myosin light chain; this was also prevented or reversed by PGE2. Remarkably, PGE2 alone caused an increase in the rate of [32P]orthophosphate incorporation into myosin light chain in cells that were otherwise untreated. Phosphorylation of the light chain of myosin may be an important factor in the maintenance of stress fiber morphology. The results of this study point out complex interrelationships between microtubules, and stress fibers in the maintenance of cell shape needing further study. The model systems that are presented seem suited for such investigation. PMID- 2992292 TI - Mechanisms of vasodilation induced by vasoactive intestinal polypeptide in rabbit mesenteric artery. AB - Vasoactive intestinal polypeptide (VIP; over 10(-13) M) inhibited the norepinephrine (NE)-induced contraction evoked from the rabbit mesenteric artery. Increased concentrations of VIP (over 10(-9) M) inhibited the contractions induced by caffeine and 39 mM [K]o. However, VIP (below 10(-7) M) had no effect on the membrane potential and resistance of muscle cells. In Ca-free solution, VIP (10(-10) M) inhibited the NE-induced contraction, but the second application of NE after removal of VIP enlarged the amplitude of contraction over that in the control. Yet when 10(-9) M VIP was applied, both the first and second contractions were consistently smaller than those observed by application of 10( 10) M VIP. In Na- and Ca-free solution, repetitive applications of NE generated contractions longer than those observed in Ca-free solution. When VIP (10(-10) M) was applied once (3 min), the contraction was inhibited only once during repetitive applications of NE. VIP (over 10(-9) M) dose dependently inhibited the NE-induced contraction and had a long-lasting inhibition after washout of the tissue. In saponin-treated skinned muscles, VIP (10(-7) M) had no effect on the Ca-induced contraction or on the Ca store sites. VIP (over 10(-8) M) was about 10 times more potent than equimolar concentrations of isoproterenol in increasing the content of adenosine 3', 5'-cyclic monophosphate (cAMP). These results indicate that VIP (10(-10) M) selectively inhibits the Ca release activated by NE, and high concentrations (over 10(-9) M) would expectedly increase the Ca extrusion from cells following increase in the levels of cAMP. PMID- 2992293 TI - Attenuation of muscarinic cholinergic inhibition by islet-activating protein in the heart. AB - Experiments were carried out to investigate the influence of islet-activating protein (IAP), a pertussis toxin, on the autonomic regulation of cardiac function. In atria isolated from rats given intravenous IAP (0.125-1.0 micrograms/100 g body wt) 12-72 h prior to experiments, the negative chronotropic and inotropic actions of carbachol were attenuated in a dose- and time-dependent manner. The inhibitory action of carbachol in the absence of isoproterenol and its attenuation by IAP were not associated with any changes in tissue adenosine 3',5'-cyclic monophosphate (cAMP) levels. In addition, carbachol decreased the isoproterenol-induced elevation of cAMP levels and inhibited the positive chronotropic and inotropic responses to isoproterenol. The inhibitory action of carbachol on the isoproterenol-induced functional and cAMP responses was also reduced by the IAP treatment. The increase in tissue guanosine 3',5'-cyclic monophosphate (cGMP) levels produced by carbachol was likewise abolished by the IAP treatment. These results indicate that IAP treatment attenuates the inhibitory actions of carbachol elicited via muscarinic receptors through both cAMP-dependent and cAMP-independent subcellular processes. PMID- 2992294 TI - Parasympathetic control of transmural coronary blood flow in dogs. AB - The transmural distribution of coronary blood flow was studied during vagal stimulation in closed-chest, morphine- and alpha-chloralose-anesthetized dogs. The left main coronary artery was cannulated and perfused at constant pressure. Bradycardia during vagal stimulation was prevented by atrioventricular heart block and ventricular pacing. Beta-adrenergic receptors were blocked with propranolol (1 mg/kg iv), and aortic pressure was stabilized by means of a pressure reservoir. Regional myocardial blood flow was measured with 9-micron radioactive microspheres during vagal stimulation and during intracoronary acetylcholine infusion. Vagal stimulation increased coronary blood flow uniformly across the left ventricular wall. In contrast, intracoronary acetylcholine infusion, at a rate selected to increase total flow to the same degree, vasodilated the subendocardium more than the subepicardium, increasing the inner/outer blood flow ratio. It is concluded that both vagal activation and acetylcholine produce coronary vasodilation that is independent of left ventricular preload, afterload, and heart rate. Acetylcholine vasodilation preferentially vasodilates the subendocardium, increasing the inner/outer flow ratio, but vagal stimulation produces uniform vasodilation across the left ventricular wall. PMID- 2992295 TI - Decreased membrane fluidity and beta-adrenergic responsiveness in atherosclerotic quail. AB - The effects of increased cholesterol on erythrocyte membrane fluidity and beta adrenergic function were studied in a quail model of atherosclerosis. Birds fed a cholesterol-supplemented diet developed severe atherosclerosis and hypercholesterolemia after 6 wk. This cholesterol-enriched diet led to a markedly elevated serum cholesterol and a 26% increase in the cholesterol-to-phospholipid ratio in erythrocyte membranes. Electron paramagnetic resonance spectra measured with 5- and 12-doxyl-stearic acid spin-label probes were used to estimate the order of quail erythrocyte membranes. Membrane preparations from cholesterol-fed birds were more highly ordered near the membrane leaflet surface, as well as deeper in the membrane interior, compared with controls. beta-Adrenergic receptor stimulation of adenosine 3',5'-cyclic monophosphate accumulation was blunted in erythrocytes from the hypercholesterolemic quail. There was no change in beta receptor density or affinity in the cholesterol-enriched membranes. These studies demonstrate that cholesterol incorporation into erythrocyte membranes in vivo is associated with decreased membrane fluidity and decreased beta-adrenergic responsiveness. The atherosclerotic quail may serve as a useful model to further probe the sequelae of hypercholesterolemia on the function of integral membrane proteins. PMID- 2992296 TI - Phentolamine-resistant neurogenic constriction occurs in small arteries at higher frequencies. AB - The effect of the alpha 1, alpha 2-antagonist phentolamine (PTA) on neuromuscular transmission and exogenous norepinephrine (NE) was assessed in arteries of diminishing diameter possessing a substantial adventitiomedial junction adrenergic innervation in the rabbit ear, i.e., central ear artery (CEA), unstretched lumen diameter (ULD) approximately equal to 300 microns; main side branch (MSB) off the CEA (ULD approximately equal to 150 microns); and terminal branch (TB) off the MSB (ULD approximately equal to 75 microns). With increasing PTA concentrations, contractile responses to transmural nerve stimulation (TNS) were decreased proportionately less in TB than in MSB and CEA. PTA (4 X 10(-6) M, a competitive antagonist concentration) blocked the tetrodotoxin-sensitive TNS induced contractions of CEA segments at 2, 4, and 8 Hz. The response at 8 Hz was reduced at least 98% in MSB and 86% in TB. However, responses to 8 Hz were not abolished in MSB and TB until 2 X 10(-5) and 3 X 10(-5) M PTA, respectively. PTA (3 X 10(-5) M) possessed nonspecific depressant properties in addition to its alpha-antagonist properties. We conclude that the sympathetic nervous system influences tone through alpha-adrenoceptors in the CEA and in the MSB and TB at lower frequencies. Responses in the MSB and TB at higher frequencies of nerve stimulation are mediated predominately through alpha-receptors. If the possibility of a nonadrenergic transmitter is discounted, the possibilities that the small PTA-resistant component of the neurogenic response is due to a high concentration of NE acting on alpha-adrenoceptors and/or a high threshold site cannot be distinguished. PMID- 2992297 TI - Control of ACTH and vasopressin in neurohypophysectomized conscious dogs. AB - Adrenocorticotropin (ACTH), cortisol, and vasopressin responses to clamped decreases in blood pressure (MAP) and to ovine corticotropin-releasing factor (CRF) infusion (20 ng X kg-1 X min-1) in intact and neurohypophysectomized (NHX) conscious dogs were examined. Mean arterial blood pressure was decreased 28 mmHg by a controlled infusion of sodium nitroprusside. Hypotension induced large increases in ACTH (peak 164 +/- 25 pg/ml), cortisol (peak 12.5 +/- 2.5 micrograms/dl), and vasopressin (peak 221 +/- 64 pg/ml) in intact (n = 7) dogs. NHX (n = 7) significantly attenuated these responses to hypotension. CRF infusion induced increases in ACTH similar in intact (n = 4) and NHX (n = 4) dogs. However, cortisol responses were significantly attenuated by NHX. Interestingly, CRF infusion induced small but significant increases in vasopressin from 3.0 +/- 1.1 to 8.1 +/- 2.0 pg/ml. We conclude that NHX attenuates ACTH and vasopressin responses to hypotension and cortisol responses to CRF-induced increases in ACTH. CRF seems to stimulate vasopressin release. PMID- 2992299 TI - Does community care for the mentally ill make a difference? A tale of two cities. AB - Matched groups of schizophrenic patients in Portland, Ore., and Vancouver, B.C., were compared approximately 1 year following discharge from an index hospitalization. Whereas Vancouver boasts a rich network of accessible private services and a public mental health system that provides a model of care for the chronically mentally ill, Portland's aftercare facilities at the time of the study were limited. One year after discharge the Vancouver cohort experienced fewer readmissions, was more apt to be employed, and reported a higher level of well-being, all of which suggest that community aftercare positively affects the negative symptoms of schizophrenia. PMID- 2992298 TI - Diagnostic categorization of psychiatric disturbance in Cushing's syndrome. AB - The author obtained a longitudinal psychiatric history from 30 patients with proven Cushing's syndrome. Twenty-five (83%) of the patients met strict diagnostic criteria for an episode of affective disorder during the course of their endocrine disturbance. Twenty patients met criteria for endogenous depression and eight patients also reported an episode of mania or hypomania. Schizophrenic syndromes were not evident, although two patients met criteria for a psychotic depression. Patients frequently attempted to minimize or conceal serious psychiatric disturbance, including a suicide attempt. Typical affective syndromes occurred in patients with adrenocortical tumors as well as in those with ACTH-dependent Cushing's syndrome. PMID- 2992300 TI - Naltrexone-induced dysphoria in former opioid addicts. AB - Naltrexone treatment, used to prevent relapse among former opioid addicts, is reported to have an extraordinary rate of noncompliance. Since activation of opioid receptors produces a sense of well-being, naltrexone's blockade of these receptors might produce dysphoria, which could contribute to noncompliance among addicts under treatment. To test this hypothesis, the authors administered naltrexone to four men who had been free of opioids for 9 to 44 months using a 6 week, placebo-controlled crossover design. One subject dropped out with abstinence-like symptoms, and two others reported mild but significantly greater dysphoria during naltrexone administration. The results suggest that naltrexone may induce mild dysphoria long after addicts stop using opioids. PMID- 2992301 TI - Favorable influences of imuthiol on mouse reproduction and immune system of offspring. AB - A physiological immature immune system in newborns is a common feature frequently associated with increased susceptibility to infections. The properties of imuthiol (purified sodium diethyldithiocarbamate), an agent specifically active on the T-cell lineage, and virtually devoid of toxicity for man or animals, encouraged us to determine whether imuthiol administered to the dams could increase the immune capability of offspring without altering fecundability and birth rate. Experiments performed either in histocompatible or histoincompatible mating systems, show that chronic administration of imuthiol prior to mating and/or during pregnancy stimulated newborn mice to increased T-cell-dependent responses, without altering birth rates and growth curves in progenies. The data suggest that imuthiol has no teratogenicity or deleterious influences on mouse gametes, and might be useful to prevent immunodepression-associated infections in newborns. PMID- 2992302 TI - A partially biodegradable material device for repair and reconstruction of injured tendons. Experimental studies. AB - We assessed the applicability of a partially biodegradable synthetic material composed of polyglycolic acid (PGA) and dacron to repair or replace severely injured tendons. Adult rabbits underwent complete laceration and repair of one Achilles tendon. Group 1 (N = 8) had end to end tenorrhaphy with size 0 braided polyester suture, and Group 2 (N = 16) tendons were similarly repaired with the bicomposite designed PGA-dacron device. Group 3 (N = 16) received laceration and removal of 1 cm of tissue, and the defect was bridged with the same PGA-dacron material. No postoperative immobilization was applied. Evaluation, consisting of biomechanical testing or histologic inspection, was done at 4 and 8 weeks after tenorrhaphy. All tendons healed, and at 8 weeks the mean maximum load at failure of the repaired tendons was 32.2 +/- 3.4 kgf, 40.3 +/- 2.4 kgf, and 31.8 +/- 3.2kgf for Groups 1, 2, and 3, respectively. Values between groups were not significantly different, but all were significantly less (P less than or equal to 0.05) than the strength of the unoperated control tendons (55.8 +/- 7.2 kgf). The most noteworthy finding was the lengthening of the repaired tendons, due to scar elongation at the tenorrhaphy site, that occurred during healing as determined by a landmark placed a fixed distance proximal to the repair site. Group 1 tendons lengthened 22 mm, while Group 2 and 3 tendons increased by 12.5 mm. Histologically, fibrous tissue ingrowth into the residual dacron scaffold occurred, but it was inconsistent and inadequate in quality and quantity to be satisfactory as neotendon. The fibrous tissue did not mature or align in response to load. We concluded that the PGA-dacron material had adequate strength and physical properties to use both for primary tenorrhaphy and to bridge the tendon defect. However, in our experience this material lacks any significant advantage over other materials available or known to be under evaluation to support neotendon formation when an actual soft tissue defect exists. PMID- 2992303 TI - Electrocardiographic changes during treatment of leishmaniasis with pentavalent antimony (sodium stibogluconate). AB - Serial electrocardiograms (ECGs) were obtained during 65 courses of sodium stibogluconate treatment in 59 Kenyan patients with leishmaniasis (56 visceral and 3 cutaneous). ECG abnormalities developed during 54% of the treatment courses. The frequency with which abnormalities occurred was related to the total daily dose of antimony (Sb), increasing from 2/9 patients treated with 10 mg Sb/kg/d to 25/48 treated with 20-30 mg Sb/kg/d and 8/8 treated with 40-60 mg Sb/kg/d. The frequency with which ECG abnormalities developed was also related to the duration of treatment, increasing from 11/65 patients after 7 days to 18/44 after 15 days, 26/39 after 30 days and 11/12 after 60 days. ECG abnormalities were similar to those previously described during treatment with trivalent antimonial drugs, the most common being flattening and/or inversion of T waves. Prolongation of the corrected QT interval occurred in 13 patients, all of whom were treated for more than 30 days or with more than 20 mg Sb/kg/d. One patient died suddenly during the fourth week of treatment with 60 mg Sb/kg/d, and 2 patients died of measles after 9 or 10 days of treatment with 30 mg Sb/kg/d. QT prolongation and a concave ST segment developed in all 3 patients who died. We conclude that minor ECG abnormalities are common when sodium stibogluconate is used at doses above 20 mg Sb/kg/d for more than 15 days, and that life threatening arrhythmias may occur if very high doses are used. PMID- 2992305 TI - Human monkeypox: a newly emerged orthopoxvirus zoonosis in the tropical rain forests of Africa. AB - During the course of the recently concluded smallpox eradication program, a new human orthopoxvirus infection was discovered which is caused by monkeypox virus. The disease occurs sporadically in remote villages within tropical rain forests of West and Central Africa. The disease is rare; only 155 cases having been reported from 1970 to 1983. The symptoms and signs of human monkeypox resemble those of smallpox, differing significantly only in the occurrence of lymphadenopathy with human monkeypox disease. Of 155 cases, some 80% are believed to have resulted from infection from an as yet unknown animal reservoir; the rest occurred among unvaccinated close contacts among whom a secondary attack rate of 15% was observed. Although person-to-person spread appears to have occurred in some instances, few cases were observed in the third or fourth generation of transmission and none thereafter. Since 1982, the incidence of human monkeypox infections in Zaire has increased concomitant with an intensified surveillance program. Additional reasons which might explain the increased incidence are discussed. Further surveillance and research of this primarily zoonotic infection are warranted and are in progress. PMID- 2992304 TI - High continuous antimony therapy in two patients with unresponsive mucosal leishmaniasis. AB - The treatment of two patients with severe mucosal leishmaniasis due to Leishmania braziliensis braziliensis is described. Both patients had received much prior antimonial therapy and one had relapsed after a total dose of 2.5 g of Amphotericin B. Both patients responded to prolonged continuous Pentostam therapy at a daily dose of 20 mg Sbv/kg/day for 62 days in one case and for 85 days in the other. Pentavalent antimonials can be curative in such protracted courses in selected patients unresponsive to standard chemotherapy. PMID- 2992306 TI - Medical pathology conference. A 54-year-old man with progressive glucose intolerance. PMID- 2992307 TI - [Plasma lipids and atheromatous plaque]. PMID- 2992308 TI - The armlift test. An alternative to the headlift test for assessing recovery from neuromuscular blockade. AB - The armlift test was assessed as an alternative to the headlift test for recovery from neuromuscular blockade. Forty comparisons of inspiratory force, duration of arm-lift and duration of headlift were performed in 27 intubated patients after anaesthesia, which included the use of non-depolarising neuromuscular blockade. Patients' ability to keep one arm raised (with the elbow clear of the bed) correlated well with inspiratory force (r = 0.67). All patients capable of an armlift duration of 45 seconds or more were also found to have an inspiratory force of at least -2.5 kPa and were safely extubated. The headlift test was found to be less discerning (r = 0.34) and of 40 determinations of headlift and inspiratory force there were 18 false negatives and two false positives. PMID- 2992309 TI - [Effect of etomidate and thiopental on ACTH and cortisol levels in serum]. PMID- 2992310 TI - Propidium iodide and S1 nuclease: tools for studying DNA reassociation kinetics. AB - A method for the determination of the amount of double-stranded DNA in a reassociation mixture is described. Reassociated DNA resistant to S1 nuclease digestion is measured fluorometrically using propidium iodide. A direct comparison is made between this method and an established method in which radiolabeled Escherichia coli DNA resistant to S1 digestion is measured by scintillation counting after separation of nucleotides by Sephadex G-100 chromatography. Reassociation curves determined for calf thymus and E. coli DNA are presented. PMID- 2992311 TI - Assay of aminoglycoside phosphotransferase in situ. AB - A method for the evaluation of the aminoglycoside phosphotransferase activity in bacterial colonies directly is described. The method is based on the ability of the enzyme to modify the substrate immobilized on carboxymethylcellulose paper. The sensitivity and accuracy of the method were tested by comparing the results of the present assay to those obtained with conventional procedures. The method seems to be particularly useful for the detection within a bacterial population producing aminoglycoside phosphotransferase of those cells which do not make the enzyme and for rapid determination of the relative levels of enzyme produced by different clones. PMID- 2992312 TI - Periodate oxidation products of hydroxylysine in the synthesis of 5-substituted prolines. AB - The amino acid hydroxylysine was subjected to oxidation by sodium metaperiodate under various conditions. It was found that in acid and high temperature, the initial oxidation product alpha-aminoglutaric gamma-semialdehyde was converted to glutamic acid with a yield of 60%. The use of alkaline conditions of oxidation favored the cyclization of alpha-aminoglutaric gamma-semialdehyde to form delta 1 pyrroline 5-carboxylic acid. Addition of NaCN to this intermediate generated new proline analogs, likely a mixture of cis- and trans-5-cyanoprolines, with a yield of 30%. Upon hydrolysis, the 5-cyanoprolines were converted to a probable mixture of cis- and trans-5-carboxyprolines. Infrared and high-resolution mass spectral data of the analogs and visual absorption spectra of the ninhydrin products were obtained to confirm the structures. PMID- 2992313 TI - An improved DNA sequencing strategy. AB - A modification of Hong's systematic DNA sequencing strategy is described. The original procedure has been simplified and transfectant yield increased. After DNase I limited cleavage in the presence of Mn2+, the single-cut linear DNA does not have to be separated from supercoiled or open circular DNA on an agarose gel. After ligation, the DNA is digested with a second restriction endonuclease for which a unique cleavage site resides between the insert and the first restriction endonuclease cutting site. The original intact DNA is linearized whereas the deleted subclone is not. The background is decreased to an undetectable level. This DNA sequencing strategy was tested on a 1.4-kb DNA fragment containing the araC regulatory gene from Erwinia carotovora. A set of subclones sufficient to sequence the fragment on both strands was produced in 2 days and the yield was at least 60-fold higher than in the original protocol. PMID- 2992314 TI - Protein hydrogen exchange studied by the fragment separation method. AB - The potential of hydrogen-exchange studies for providing detailed information on protein structure and structural dynamics has not yet been realized, largely because of the continuing inability to correlate measured exchange behavior with the parts of a protein that generate that behavior. J. Rosa and F. M. Richards (1979, J. Mol. Biol. 133, 399-416) pioneered a promising approach to this problem in which tritium label at exchangeable proton sites can be located by fragmenting the protein, separating the fragments, and measuring the label carried by each fragment. However, severe losses of tritium label during the fragment separation steps have so far rendered the results ambiguous. This paper describes methods that minimize losses of tritium label during the fragment separation steps and correct for losses that do occur so that the label can be unambiguously located and even quantified. Steps that promote adequate fragment isolation are also described. PMID- 2992315 TI - A high-performance liquid chromatographic technique that separates cellular retinol binding protein from cellular retinoic acid binding protein. AB - A one-step procedure to detect cellular [3H]retinol and [3H]retinoic acid binding proteins (CRBP and CRABP) from rat testis cytosolic extract was devised. The procedure is based on anion-exchange high-performance liquid chromatography of the cytosolic fraction on columns of Mono Q, which permits elution of CRABP and CRBP at 12 and 22 min, respectively. PMID- 2992316 TI - The relationship of evoked electromyographic and mechanical responses following atracurium in humans. PMID- 2992317 TI - Replication of tick-borne encephalitis (TBE) virus in ticks Dermacentor marginatus. AB - In the laboratory experiments, the virophoric period in D. marginatus ticks lasted 61 to 81 d, the premoulting period (nymphs - adults) amounted to 17 to 18 d. The titres of individually examined females and males for the presence of TBE virus ranged from 10(1) to 10(5.5) ic mouse LD50/0.03 ml between the 14th and 29th day after hatching. In the laboratory experiments 77% of ticks were positive. In the field experiments, the virophoric period in D. marginatus ticks lasted 79 to 114 d, the premoulting period amounted to 34 to 38 d. The titres of individually examined adults for the presence of virus ranged from 10(1) to 10(5.5) ic mouse LD50/0.03 ml between the 13th and 80th day after hatching. In the field experiments 96.5% of ticks were positive. PMID- 2992318 TI - Coxsackievirus B4 infection of sympathetic ganglia in squirrel monkeys. AB - Lumbar sympathetic ganglionitis was found by light microscopy in 2 of 17 (12%) squirrel monkeys (Saimiri sciureus) experimentally infected with Coxsackievirus B4. This finding shows that viruses can cause ganglionitis which, in turn, must cause autonomic nervous system dysfunction. Such viral ganglionitis may explain some diseases, including cardiovascular ones, of poorly understood or unknown etiology which present with manifestations of dysfunction of the sympathetic nervous system. PMID- 2992319 TI - Superior vena caval obstruction: an analysis of seventy-six cases, with comments on the safety of venography. AB - Seventy-six cases of superior vena caval obstruction (SVCO) were documented in the period 1970-1980. There were 53 males and 23 females, with a mean age of 61.7 years. The underlying causes were:- Lung cancer, 64/76, 84.2%; Metastatic disease, 4/76, 5.3%; Lymphoma, 4/76, 5.3%; Benign aetiology, 2/76, 2.6%; Undiagnosed, 2/76, 2.6%; 97.4% of the cases were due to malignancy. Of 61 patients on whom follow-up is available, only one is alive. Eighty-two percent of this group died within one year following the onset of SVCO. Autopsies were performed on 17 patients. Nine of these (52.9%) showed invasion of the superior vena cava, 6 (35.3%) showed compression without invasion, and 2 (11.8%) showed neither compression nor invasion following Radiotherapy. Venography was performed on all patients with only one minor complication being recorded. PMID- 2992320 TI - Effect of human leukocyte A interferon on prevention of infectious bovine rhinotracheitis virus infection of cattle. AB - Human leukocyte A interferon was evaluated for its ability to prevent infectious bovine rhinotracheitis virus-induced respiratory tract disease in cattle. Weanling calves were treated daily for 1 week with 50 X 10(6) U of interferon, intranasally (by nebulization) and IM, and inoculated with infectious bovine rhinotracheitis virus on the first day of treatment. Respiratory tract disease was less severe in treated as compared with nontreated calves which were given only infectious bovine rhinotracheitis virus, and infection in the treated calves occurred later than in the untreated calves. Viral shedding and appearance of viral neutralizing antibodies occurred later in treated calves than in calves given only virus. Because several calves in a treatment group were housed together, whether the late appearance of infection in some interferon-treated calves was due to emergence of suppressed virus or to horizontal transmission from calves shedding virus could not be determined. One calf in the interferon treated group developed antibody to human interferon and a few treated calves had transient elevation of hepatic enzymes. Interferon-treated calves developed a high temperature which subsided on termination of treatment. Production of disease was considerably dependent on the amount of virus and interferon given, since calves given 300 times more virus and approximately half as much interferon showed no evidence of protection against infection. PMID- 2992321 TI - Abnormal canine triiodothyronine-binding factor characterized as a possible triiodothyronine autoantibody. AB - Of more than 6,000 canine serum samples submitted to the Endocrine Diagnostic Laboratory of the School of Veterinary Medicine, Auburn University, Ala, for thyroid evaluation in 1983, 18 contained an abnormal triiodothyronine (T3) binding factor (T3BF). These samples were easily distinguished from non-T3BF containing samples because the factor interfered with the radioimmunoassay for total T3 resulting in a profound increase in apparent values for T3 concentration. Most of the T3BF-containing samples did not have unusual thyroxine binding or inappropriate thyroxine concentrations. Triiodothyronine binding in samples containing T3BF was inhibited by addition of exogenous T3 and was, with one exception, not affected by the addition of 8-anilino-1-naphthalene sulfonic acid (ANS). Apparent affinity constants determined by Scatchard analysis were considerably greater than those of circulating T3-binding proteins found in human beings, but were similar to values obtained for human T3 autoantibodies. After the addition and incubation of [125I] T3 with T3BF-containing samples, a large fraction of the radioactivity bound by T3BF could be precipitated by the addition of goat antibody to dog immunoglobulin G. Additional observations that the binding was insensitive to inhibition by ANS, was resistant to heat inactivation, and had a high apparent affinity constant for T3 were consistent with the idea that the factor was an autoantibody to T3. PMID- 2992322 TI - Attenuated properties of thymidine kinase-negative deletion mutant of pseudorabies virus. AB - A thymidine kinase (TK)-negative (TK-) deletion mutant of the Bucharest (BUK) strain of pseudorabies virus (PRV) was isolated. The mutant, designated as PRV (BUK d13), did not revert to TK-positive (TK+), even when propagated in medium that selected for TK+ viruses. The mutant also replicated equally well at 39.1 C and 34.5 C, and was easily distinguished from other PRV strains by molecular hybridization experiments, restriction nuclease fingerprints, and plaque autoradiography or other assays for the TK phenotype. The PRV (BUK d13) had greatly reduced virulence for mice and rabbits, compared with parental TK+ strains, PRV (BUK-5) and PRV (BUK-5A-R1), and provided mice with solid protection against the TK+ BUK and Aujeszky strains of PRV. Experiments were done in 5- to 6 week-old pigs to assess the safety and efficacy of PRV (BUK d13) in the natural host. In one experiment, pigs were vaccinated IM with 7.5 X 10(8) plaque-forming units of TK- PRV (BUK d13), and were then challenge exposed intranasally (IN) with 4.3 X 10(8) TCID50 of virulent PRV [Indiana-Funkhauser (IND-F)]. Vaccinated pigs did not have clinical signs of illness after vaccination or after challenge exposure. One nonvaccinated control pig died on postchallenge day 4; a 2nd nonvaccinated control pig became moribund, but eventually recovered. Pigs developed virus-neutralizing antibodies after vaccination, and had a secondary immunologic response after challenge exposure; however, PRV was not isolated from the tonsils or trigeminal ganglia of vaccinated pigs at postchallenge exposure day 11.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992323 TI - Retrospective study of four years of carcass condemnation rates for malignant lymphoma in California cows. AB - A retrospective study was undertaken to examine monthly rates of carcass condemnation for bovine malignant lymphoma in adult dairy cows slaughtered between January 1979 and December 1982, in 2 plants (A and B) in the southern San Joaquin Valley of California. Extremes (mean) of monthly rates for slaughterhouse A were 41 to 100 (65.9) per 10,000 slaughtered and for slaughterhouse B, 39 to 113 (94.1) per 10,000. The overall monthly mean rate was 80 per 10,000 slaughtered. Equations for long-term trend lines for condemnation rates for slaughterhouses A and B were T = 0.398 + 0.0176t - 0.0002t2 and T = 0.314 + 0.0378t - 0.0007t2, where t = month. A difference in patterns of trends was not apparent, although slaughterhouse A tended to have lower rates of condemnation than did slaughterhouse B. Seasonal components of trend lines, estimated by time series analysis, were not consistent between plants, except for the month of August, during which time seasonally adjusted rates were low for both slaughterhouses. For slaughterhouse A, 2 classic cyclical components were identified--between January 1981 and September 1981 and between September 1981 and September 1982. For slaughterhouse B, the cyclical components were between November 1979 and July 1981 and between July 1981 and August 1982. The later cycle resembled the second cycle of rates from slaughterhouse A. Rates remained stable through 1979 and then increased steadily for 2.5 years. During the last 6 months of 1982, rates leveled off and perhaps began to decline. PMID- 2992324 TI - Comparison of the commercial agar-gel immunodiffusion test and radioimmunoprecipitation assay for detection of antibodies to bovine leukemia virus. AB - In a double-blind study, the commercial agar-gel immunodiffusion test (AGID) was compared with a radioimmunoprecipitation assay (RIA) performed with glycoprotein (gp) antigen for detection of antibodies to bovine leukemia virus. Of 240 sera tested, 115 were from adult cows and 125 were from precolostral calves. Most adult animals were tested within 1 week of parturition. Sera from 74 cattle were positive and sera from 166 cattle were negative by gp RIA. Sensitivity of the AGID, compared with the gp RIA, was 85.1% when the test was read at 48 hours and was 94.6% when read at 72 hours. Specificity increased from 92.2% at 48 hours to 96.4% at 72 hours. Reading the AGID again at 72 hours also clarified most reactions that were questionable at 48 hours due to a haze around the test serum well. Of 3 RIA-positive precolostral calf sera, 2 were AGID-negative and 1 had a questionable reaction by the AGID at 48 hours. Of 5 RIA-positive sera that were AGID-negative at 48 hours, 2 were precolostral calves and 3 were cows tested at parturition. Of 166 RIA-negative reactions, none was falsely positive by the AGID at 48 or at 72 hours. PMID- 2992325 TI - Effect of primary and recurrent infections bovine rhinotracheitis virus infection on the bovine ovary. AB - Six heifers were inoculated IV at estrus with the Iowa or Colorado isolates of infectious bovine rhinotracheitis virus (IBRV). Subsequent measurements of plasma progesterone indicated that corpus luteum function was depressed in all heifers. In the 1st estrous cycle after inoculation, progesterone values did not exceed 2 ng/ml in 3 heifers given the Iowa isolate. Although maximal progesterone values were greater than or equal to 2 ng/ml in 3 heifers given the Colorado isolate, values were lower than those in later cycles. Five heifers had maximal diestrual progesterone values greater than or equal to 5 ng/ml within 5 weeks after inoculation, but in the 6th heifer, this amount of progesterone was not present until 8 weeks after inoculation. Three to 5 months after inoculation, all heifers were given 5 daily injections of dexamethasone, 2 heifers each during metestrus, diestrus, or proestrus. Subsequent recrudescence of IBRV was demonstrated in all heifers by the isolation of virus from vaginal or nasal swab samples. The heifers were killed 10 to 17 days after initiation of dexamethasone treatment and their reproductive organs were examined for lesions and IBRV. Lesions were not seen, and IBRV was isolated only from the corpus luteum of a heifer given dexamethasone during diestrus. PMID- 2992326 TI - Vaccination of swine with thymidine kinase-deficient mutants of pseudorabies virus. AB - A mutant of pseudorabies virus (PRV) deficient in thymidine kinase (TK-) activity was isolated and characterized. The mutant grew well in cell culture and did not revert to the thymidine kinase-positive phenotype. The PRV-TK- was not virulent when inoculated intranasally into 3-to 4-week-old pigs and could not be reactivated from the ganglia of these pigs by explantation and cocultivation with susceptible cells several weeks after virus inoculation. Pigs that had been exposed to PRV-TK- were immune to challenge exposure with a virulent strain of PRV. Furthermore, the challenge virus was not recovered from the ganglia of most of these pigs, indicating that colonization of the ganglia by a super-infecting virulent PRV strain was considerably reduced by vaccination. PMID- 2992327 TI - Experimental infection of fattening pigs with pseudorabies (Aujeszky's disease) virus: efficacy of attenuated live- and inactivated-virus vaccines in pigs with or without passive immunity. AB - The efficacies of attenuated live- and inactivated-virus vaccines against pseudorabies (PR) in fattening pigs were compared. Pigs born from vaccinated or nonvaccinated sows were vaccinated with one or the other vaccine and were challenge exposed at the end of the fattening period. The particular form of PR observed in fattening units in the field could be reproduced. A marked difference was seen between the control lot and the lots of the pigs vaccinated with the attenuated live- and inactivated-virus vaccines. The protection was real, but not absolute, in the vaccinated pigs. The inactivated-virus vaccine conferred a strong passive immunity to the young pigs of vaccinated dams which interfered with the development of an active immunity. The titer of the colostral antibodies in the sera of pigs born from the sows vaccinated with the attenuated-live virus vaccine was low and decreased rapidly. The active protection obtained with this vaccine was similar to that observed with the inactivated-virus vaccine. Thermal curves, weight losses, and time necessary for recovering the weight of pigs at the time of challenge exposure seemed to be good criteria for measuring the protection of fattening pigs against this particular form of PR. The conditions of the outcome of respiratory tract disorders in these pigs are discussed. PMID- 2992328 TI - Pathologic changes in pigs with prednisolone-induced recrudescence of herpesvirus infection. AB - In pigs inoculated with Aujeszky's disease virus (ADV), recrudescence of herpesvirus infection was induced by daily administration of 1,000 mg of prednisolone for 5 days at 2 (group A) or 5 (group B) months after the primary infection. At necropsy in group A pigs, ADV was recovered from nasal secretions 3 to 9 days after prednisolone treatment initiation and from the brain cortex 10 days after treatment initiation; ADV was not recovered from group B pigs. In pigs of both groups killed 10 days after treatment initiation, 2 types of characteristic lesions were found. One type was a nonsuppurative encephalitis that consisted of neuronal necrosis, neuronophagia, and formation of eosinophilic intranuclear inclusion bodies. The 2nd type had basophilic intranuclear inclusion bodies in the enlarged endothelial cells of the kidneys, liver, lungs, adrenal glands, and lymph node sinusoids. Cells containing intranuclear inclusion bodies had immature and mature herpesvirus particles. Therefore, the brain lesions containing the eosinophilic inclusion bodies were considered to be due to ADV. Basophilic inclusion bodies in the endothelial cells were due to porcine cytomegalovirus. These observations indicated that prednisolone treatment resulted in recrudescence of ADV and porcine cytomegalovirus infections. PMID- 2992329 TI - Effect of o,p'DDD therapy on endogenous ACTH concentrations in dogs with hypophysis-dependent hyperadrenocorticism. AB - The effect of o,p'DDD therapy on the endogenous plasma ACTH concentration was evaluated in 15 dogs with hypophysis-dependent hyperadrenocorticism. Adequate control of hyperadrenocorticism with o,p'DDD was based on the reduction of water consumption to within the normal range, disappearance of clinical signs of lethargy, weakness, alopecia, thin skin, or pendulous abdomen, and an increase in blood cortisol below the normal range after exogenous ACTH administration. Endogenous ACTH concentrations were determined for each dog after the disease was controlled and while they were given o,p'DDD on a maintenance schedule. Endogenous ACTH concentrations increased in 14 of 15 dogs after o,p'DDD therapy, indicating a lack of suppressive effects of o,p'DDD on hypophysis ACTH secreting cells. PMID- 2992331 TI - In vivo and in vitro studies on the use of the guinea pig as a model for virus provoked airway hyperreactivity. AB - The parainfluenza 3 (P-3)-infected guinea pig was examined as a model for virus provoked airway hyperreactivity by measuring changes in airway overflow pressure in response to intravenously (iv) administered histamine. In vitro responses of lung parenchymal strips to several contractile substances were also measured. All studies were conducted 4 days after nasal insufflation with P-3 or P-3 growth medium (control). Increases in airway overflow pressure caused by histamine were enhanced by P-3 infection, and the dose required to produce a standard level of response was decreased (i.e., there was an increase in sensitivity to histamine). Enhancement of in vivo histamine responses caused by P-3 was prevented both by cutting the vagus nerves in the midcervical region and by iv administered hexamethonium, 5 mg/kg. The enhancement was not blocked by 1 mg/kg of atropine given iv, but was blocked by a larger dose 5 mg/kg. The larger dose of atropine significantly antagonized responses to histamine in the P-3-infected state. Increases in airway overflow pressure produced by electrical stimulation of the left vagus and nicotine (1 and 10 mg/kg given iv), both studied after bilateral vagotomy and propranolol, 1 mg/kg given iv, were also enhanced by P-3 infection. Atropine, 1 mg/kg given iv blocked the P-3-induced enhancement of responses to vagus stimulation. Propranolol, 1 mg/kg, and phentolamine, 3 mg/kg, given together iv, produced a doubling of the airway overflow pressure only in P-3 infected animals. Propranolol alone or other receptor antagonists did not produce as marked a change in either group of animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992330 TI - Effect of exogenous ACTH on serum corticosterone and cortisol concentrations in the Moluccan cockatoo (Cacatua moluccensis). AB - The effects of exogenous adrenocorticortrophic hormone (ACTH) on the serum corticosterone and cortisol concentrations were determined in 28 mature Moluccan cockatoos (Cacatua moluccensis), a representative of the psittacine species. Birds were randomly assigned to 4 groups (2 ACTH-treated groups and 2 saline treated controls). Group I (10 cockatoos [5 males and 5 females] ) was given 15 IU of ACTH after blood samples (base line) were taken at 10:00 AM. Blood samples were taken again at 30 minutes and 2.5 hours after ACTH administration. Group II (10 cockatoos) was given similar treatment, but blood samples were taken at 1 and 4 hours after ACTH was administered. Groups III and IV (each of 4 birds) were given saline solution injections as controls. Blood samples were taken at 30 minutes and 2.5 hours after injection (group III) and at 1 and 4 hours after injection (group IV). All serum samples were analyzed for cortisol and corticosterone. Serum corticosterone concentration increased significantly (P less than 0.01) from base-line levels (26 ng/ml) to 108 ng/ml within 30 minutes after ACTH was administered. The high values were maintained for 3 hours and then decreased to 40 ng/ml at the end of 4 hours. Male birds seemed to respond to the ACTH treatment quickly and maintained increased concentration for a shorter period when compared with the responses seen in female birds. Serum cortisol values remained low throughout the experimental period. These results indicate that serum corticosterone was responsive to ACTH administration, but cortisol was not. In addition, there may be a difference in the responses between male and female members of the species. PMID- 2992332 TI - Neuropsychiatric dysfunction in primary Sjogren's syndrome. AB - Neuropsychiatric complications developed in 40 patients with primary Sjogren's syndrome, none of whom met American Rheumatologic Association criteria for systemic lupus erythematosus. Twenty-five patients had psychiatric abnormalities, the commonest of which were affective disturbances. Of 30 patients tested, 23 had an abnormal pattern in the Minnesota Multiphasic Personality Inventory, the commonest pattern being a "conversion V." In general, patients presented with hysteroid dysphoric features. Of 16 patients undergoing cognitive function testing, 7 showed mild memory impairment with attention and concentration deficits. On clinical evaluation, 27 patients had neurologic abnormalities unattributable to other causes (central and peripheral nervous system in 16 and 19 patients respectively). There was a significant correlation between psychiatric disturbances and neurologic dysfunction, suggesting a possible organic basis for psychiatric dysfunction. The diagnosis of primary Sjogren's syndrome should be considered in patients with unexplained neuropsychiatric illness. PMID- 2992333 TI - Activity of 9-[2-hydroxy-1-(hydroxymethyl)ethoxymethyl]guanine in the treatment of cytomegalovirus pneumonia. AB - Ten marrow transplant recipients with biopsy-proven cytomegalovirus pneumonia were treated with the acyclic nucleoside analog 9-[2-hydroxy-1-(hydroxymethyl) ethoxymethyl]guanine (BW B759U). Viruria and viremia ceased after 4 days of treatment in all patients with cultures initially positive from these sites. Cytomegalovirus was eliminated from respiratory secretions after a median of 8 days. Despite this antiviral effect, only one patient survived the pneumonia. Quantitative cultures of lung tissue before and after treatment confirmed that therapy with BW B759U was associated with substantial antiviral activity, with a mean decrease in viral titers of more than 99.99% after treatment. Neutropenia developed in three patients when mean peak and trough plasma levels exceeded 50 and 10 mu mol/L, respectively, but no other toxicity was seen. BW B759U is the first antiviral agent showing consistent activity against cytomegalovirus in vivo, and it should be evaluated in the earlier management of cytomegalovirus infections after marrow transplantation and in serious cytomegalovirus infections in other immunocompromised patients. PMID- 2992335 TI - 9-(1,3-Dihydroxy-2-propoxymethyl)guanine for cytomegalovirus infections in patients with the acquired immunodeficiency syndrome. PMID- 2992334 TI - Treatment of cytomegalovirus retinitis with 9-[2-hydroxy-1 (hydroxymethyl)ethoxymethyl]guanine. AB - Cytomegalovirus infections can cause significant morbidity and mortality in immunosuppressed patients, and present antiviral agents have had little efficacy against these infections. We describe the use of an acyclic nucleoside 9-[2 hydroxy-1-(hydroxymethyl) ethoxy methyl]guanine (BW B759U), in the treatment of two patients with cytomegalovirus retinitis. The efficacy of this agent was evident, with healing of retinal lesions and resolution of viremia and viral shedding. BW B759U appears to be similar pharmacokinetically to intravenous acyclovir in terms of half-life, peak serum levels, and renal excretion. PMID- 2992336 TI - Revision of the case definition of acquired immunodeficiency syndrome for national reporting--United States. Centers for Disease Control, Department of Health and Human Services. AB - Since the first case definition of the acquired immunodeficiency syndrome was published in 1982, minor revisions have been made in the list of diseases used as indicators of underlying cellular immunodeficiency, and, more recently, the human T-lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV) has been identified as the cause of the syndrome. To reflect these changes, the Conference of State and Territorial Epidemiologists suggested resolutions to the case definition of the syndrome that the Centers for Disease Control will immediately adopt. These revisions will result in the reclassification of less than 1% of cases previously reported. PMID- 2992338 TI - Extrahepatic biliary obstruction and small-cell lung carcinoma. PMID- 2992337 TI - Patients with small-cell lung cancer treated with combination chemotherapy with or without irradiation. Data on potential cures, chronic toxicities, and late relapses after a five- to eleven-year follow-up. AB - We assessed the outcome in 252 patients with small-cell lung cancer 5 to 11 years after treatment with combination chemotherapy, with or without chest and cranial irradiation, in National Cancer Institute therapeutic trials from 1973 through 1978. Twenty-eight patients (11%) survived free of cancer for 30 months or more. Fourteen patients remain alive without evidence of cancer beyond 5 years (range, 6.4 to 11.3 years), and 7 patients have returned to a lifestyle similar to that before diagnosis. The other 14 patients who were cancer-free at 30 months have developed cancer or died; 6 patients had a relapse, 4 developed or died from non small-cell lung cancer, and 4 died of unrelated causes. A few patients with small cell lung cancer (5.6%) may be cured. Thirty-month, cancer-free survival is insufficient to show a cure. Although late toxicities are troublesome, they do not outweigh the benefits of prolonged survival and potential for cure with modern aggressive therapy in small-cell lung cancer. PMID- 2992339 TI - Oat-cell carcinoma in transfusion-associated acquired immunodeficiency syndrome. PMID- 2992340 TI - [Purine metabolism and blastogenesis in lymphocytes of alcoholic subjects]. AB - Immune deficiency is characteristic of alcoholic subjects. These subjects usually show altered lymphocyte function. We determined the activities of adenosine deaminase (ADA) and ecto-5'-nucleotidase (ecto-5'N) in lymphocytes from 54 subjects: 15 healthy controls, 28 non-cirrhotic alcoholics, 8 alcoholic cirrhotics and 3 non-alcoholic cirrhotics. Whereas ADA activity was the same for all 54 subjects, ecto-5'N activity was in general lower in alcoholic subjects after cessation of alcohol intake. Following alcohol intoxication, however, ecto 5'N activity increased. The decrease of ecto-5'N activity in alcoholic subjects might be explained by shedding of the ecto-enzyme and alteration of lymphocyte subpopulations. We observed decreased mitogenic-induced lymphoblastic transformation in 3 patients with cirrhosis. All other subjects (including healthy controls) had normal mitogenic-induced blastogenesis. Interestingly, following alcohol intake, non-stimulated lymphoblastic transformation increased, leading to an apparently decreased stimulation index. PMID- 2992342 TI - Cholecystokinin in the medial parvocellular subdivision of the paraventricular nucleus. Co-existence with corticotropin-releasing hormone. PMID- 2992341 TI - [Increased serum IgA levels in alcoholic polyneuropathy. Pathogenetic discussion]. AB - Increased serum IgA in patients with direct toxic alcoholic peripheral neuropathy was studied prospectively in 33 patients with an obvious clinical polyneuropathy confirmed by electrophysiological investigations. The average concentration of IgA in this group (450 mg/100 ml) was significantly higher than in a group of 24 matched alcoholic control patients without peripheral neuropathy (306 mg/100 ml), p less than 0.0001. The authors discuss possible direct and indirect modifications of the humoral immune response due to the peripheral neuropathy. PMID- 2992343 TI - Cholecystokinin in the retina of vertebrates. PMID- 2992344 TI - Integrated anatomical and physiological studies of neuronal cholecystokinin receptors. PMID- 2992345 TI - Cholecystokinin receptors in mammalian brain. A comparative characterization and visualization. PMID- 2992346 TI - Brain cholecystokinin receptors. Binding characteristics, covalent cross-linking, and evolutionary aspects. PMID- 2992347 TI - Cholecystokinin and cultured spinal neurons. Immunohistochemistry, receptor binding, and neurophysiology. PMID- 2992349 TI - Reduced cholecystokinin levels in the limbic lobe in schizophrenia. A marker for pathology underlying the defect state? PMID- 2992348 TI - Central nervous system cholecystokinin and the control of feeding. AB - There is still much to learn of the sites and mechanisms of action of brain CCK peptides and their interaction with other brain peptides and neurotransmitters. The findings obtained from studies in sheep and other species provide evidence for brain CCK peptides functioning as important transmitters of hunger and satiety signals. We have hypothesized that signals, either neural or humoral, peripherally generated as a result of feeding induce secretion of CCK from specific (paraventricular) brain sites into the CSF (FIG. 1). Specialized ependymal cells, such as tanycytes, may take up CCK from the CSF for transport to receptor sites which mediate CCK's specific functions (such as changes in rumen motility, suppression of insulin secretion, and behavioral satiety). Evidence also exists to indicate the involvement of specific hypothalamic sites in either the release or the action of CCK. PMID- 2992351 TI - A membrane filter technique for glass fibres. PMID- 2992350 TI - Studies on the conformation, enzymatic degradation, pharmacological potency, and binding properties in brain tissue of cholecystokinin-8 and new related peptides. PMID- 2992352 TI - Determination of free quartz surfaces in coal mine dust. PMID- 2992353 TI - Hearing loss in experimental cytomegalovirus infection of the guinea pig inner ear: prevention by systemic immunity. AB - Guinea pig cytomegalovirus (GPCMV) has been used to establish a reproducible model of viral labyrinthitis and hearing loss. Cochlear function was assessed by electrophysiological recordings of cochlear microphonic (CM) and eighth nerve N1 compound action potential (AP) thresholds prior to and up to eight days following inoculation of the scala tympani. Inner ear inoculation of seronegative subjects with live GPCMV produced profound elevations in CM and AP thresholds: 70% of these subjects had their thresholds raised to the limits of the sound system throughout the tested frequency range of 0.10 to 32 kHz. Histopathologic effects associated with CM and AP threshold shifts were primarily limited to the perilabyrinthine compartment, and were greatest in the most basal cochlear turns. Systemic infection with GPCMV produced an immune response, but did not affect CM or AP thresholds. Subsequent inoculation of the inner ear of these seropositive animals with live GPCMV did not result in either CM or AP threshold shifts, or cochlear histopathology. Inoculations of inactivated virus into the inner ears of seronegative and seropositive animals produced only moderate CM and AP threshold effects. Primary GPCMV labyrinthitis thus results in significant cochlear dysfunction and histopathologic changes which are prevented by prior systemic infection with GPCMV. PMID- 2992354 TI - Acute appendicitis related to faecal stasis. AB - The objectives of the study presented here were to evaluate the role of faecal stasis in the development of acute appendicitis. Inquiries about bowel habits were made of 188 patients admitted to this Department of Surgery with a clinical diagnosis of acute appendicitis, and of 188 healthy volunteers without known gastrointestinal disease, matched for age and sex. The results show that the number of bowel actions per week were significantly lower in the group of patients with acute appendicitis. Furthermore, the patients with a gangrenous and perforated appendix found at operation had significantly fewer bowel actions per week than those with phlegmonous appendicitis. These findings support the hypothesis of the importance of the obstruction of the appendix lumen in the development of acute appendicitis. PMID- 2992355 TI - A direct radioimmunoassay for the determination of adrenocorticotropic hormone (ACTH) and a clinical evaluation. AB - A direct radioimmunoassay for the determination of ACTH in a small volume of serum was developed using a commercially available antiserum and tracer. A single stage radioimmunoassay with an incubation period of 4 days gave reliable and reproducible ACTH values. In the incubation medium 0.1% 2-mercaptoethanol was used as the antioxidant. During a 15-month period the assay has been used routinely in the laboratory. The coefficient of variation was less than 10% in the physiological and pathological range. A normal range of 12 to 60 ng/L was found. The reliability of the assay is demonstrated by results obtained in patients with disorders of the pituitary-adrenal axis. PMID- 2992356 TI - Action of Lambert-Eaton myasthenic syndrome IgG at mouse motor nerve terminals. AB - We have studied the electrophysiological effects of IgG obtained from four patients with Lambert-Eaton myasthenic syndrome (LEMS) (two with small cell carcinoma), using the mouse passive transfer model. Mice received LEMS or control IgG or plasma, 10 to 60 mg daily. Microelectrode intracellular recordings were made from diaphragm muscle. LEMS IgG and plasma decreased end-plate potential quantal content similarly, confirming IgG as the active factor. LEMS IgG was equally effective in C5-deficient mice, indicating that late complement components are not required. The time course of decline and recovery of quantal content closely followed that of the human IgG in the mouse serum, with time to half-maximal effect of about 1.5 days in each case. Binding/dissociation of IgG or down/up regulation of the antigenic determinants, possibly Ca2+ channels, has a half-life of between 2 and 36 hours. The results confirm our concept that IgG antibody to nerve terminal determinants underlies the disorder of transmitter release in LEMS. PMID- 2992357 TI - Receptors and phosphoinositide-generated second messengers. PMID- 2992358 TI - Growth hormone releasing factors. PMID- 2992359 TI - The chemical modification of enzymatic specificity. AB - The chemical modification of enzymes has played and will continue to play an important role in probing the mechanism of enzyme activity. This technique can be utilized for identification of those individual amino acid residues responsible for the catalytic properties of the entire protein. In chemical modification experiments, changes in enzymatic specificity have been noted, but often not predicted. However, in recent years, rational approaches for the alteration of enzymatic properties have become feasible by means of site-specific mutagenesis and chemical methodology. In the first method, one amino acid can be replaced by a new one; in the second method, not only can new amino acid residues be introduced but also new catalytic entities (such as flavins) can be affixed to the protein molecule. Both methodologies are in their infancy, yet they represent a potentially powerful approach toward the design and synthesis of enzymes possessing new specificities. PMID- 2992360 TI - DNA topoisomerases. PMID- 2992361 TI - Transpositional recombination in prokaryotes. PMID- 2992362 TI - Protein-tyrosine kinases. PMID- 2992363 TI - [Aminoglycoside acetyltransferase from Streptomyces fradiae--a producer of neomycin]. AB - An enzyme inactivating aminoglycosides by acetylation was isolated from S. fradiae 918 producing neomycin. By the data on the substrate specificity the enzyme was identified as aminoglycoside-3-acetyltransferase. Localization of aminoglycoside-3-acetyltransferase in the cells of the culture was determined. The enzyme could be detected in the spores and mycelium of S. fradiae already at the early hours of the fermentation process. The enzyme was not detected in the fermentation broth filtrate. The activity of aminoglycoside-acetyltransferase did not depend on the cultivation conditions providing different antibiotic production levels. PMID- 2992364 TI - [Isolation and purification of restriction endonuclease PmiI from Proteus mirabilis 1667]. AB - A new restriction endonuclease Pmi I was detected in Proteus mirabilis 1667. The enzyme hydrolyzes DNA of the phage lambda into 10 electrophoretically separating fragments with molecular weights of 1.3-7.9 mD. With the use of two-stage chromatography on blue sepharose and phosphocellulose it is possible to obtain restriction endonuclease Pmi I free of the admixtures of ballast proteins, nonspecific nucleases and phosphatases. PMID- 2992365 TI - In vitro activity of WIN 51711, a new broad-spectrum antipicornavirus drug. AB - WIN 51711 (5-[7-[4-(4,5-dihydro-2-oxazolyl)phenoxy]heptyl]-3-methylisoxazole), a new antipicornavirus drug, is a potent inhibitor of human entero- and rhinoviruses at concentrations not inhibitory to HeLa cell growth. In plaque reduction assays, WIN 51711 reduced plaque formation by 9 enteroviruses and 33 rhinoviruses, with MICs of 0.004 to 0.17 and 0.004 to 6.2 micrograms/ml, respectively. Addition of WIN 51711 to infected cells at concentrations of 0.02 to 5.0 micrograms/ml reduced the yield of picornaviruses by 90%. Other RNA viruses (nonpicornaviruses) and DNA viruses were unaffected by the compound. PMID- 2992366 TI - Plasmid-borne or chromosomally mediated resistance by Tn7 is the most common response to ubiquitous use of trimethoprim. AB - The folic acid analog trimethoprim has been in clinical use for more than 10 years. The use of it in Sweden has doubled in the last 6 to 7 years, and from the distribution statistics it can be calculated that during 1 year 4 to 5% of the population in Sweden are given this drug. The bacterial resistance mechanisms to be found in response to such a selection pressure were investigated in a relatively isolated population in northern Sweden (the county of Jamtland), in which one centrally located bacteriological laboratory serves the area. Trimethoprim-resistant strains were collected during an 8-month period from consecutive specimens of bacteria from the urinary tracts of patients. Among the highly resistant strains of enteric bacteria, trimethoprim resistance mediated by transposon-borne dihydrofolate reductase of type I was found to dominate. The corresponding Tn7-like transposon was found to be localized both on the chromosome of isolated Escherichia coli strains and also on a 50-kilobase IncI transferable plasmid which was found in several different serotypes of E. coli. In two enterobacterial strains, resistance to more than 10(3) micrograms of trimethoprim per ml was furthermore found to be caused by a ca. 80-fold increase in the formation of chromosomal dihydrofolate reductase. PMID- 2992368 TI - Enhanced efficacy of the acyclic nucleoside 9-(1,3-dihydroxy-2 propoxymethyl)guanine in combination with gamma interferon against herpes simplex virus type 2 in mice. AB - The acyclic nucleoside 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) and recombinant mouse interferon gamma (rMuIFN-gamma) were evaluated for their efficacy alone and in combination against a herpes simplex virus type 2 systemic infection in mice. Intraperitoneally infected animals were treated once a day with the drugs at various concentrations for 5 days starting 24 h after inoculation. DHPG was given subcutaneously and rMuIFN-gamma intraperitoneally. For DHPG, the effective dose at which 50% of the mice survived (ED50 was lowered approximately 10-fold from 3.4 to 0.25 mg/kg when given in combination with an ineffective dose of 4MuIFN-gamma (10(3) units per mouse). For rMuIFN-gamma, the ED50 was lowered greater than 10-fold from 6 x 10(3) to less than 3 x 10(2) units per mouse when given in combination with a marginally effective dose of DHPG (1 mg/kg). Construction of an isobologram and calculation of the corresponding fractional protective dose index (less than 0.12 where values less than or equal to 0.5 are considered synergistic) indicates an enhanced protective interaction by the combination of the two drugs. PMID- 2992367 TI - Comparative efficacy and selectivity of some nucleoside analogs against Epstein Barr virus. AB - The effects of (2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodocytosine (FIAC), 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-methyluridine (FMAU), 1-(2-deoxy-2 fluoro-beta-D-arabinofuranosyl)-5-iodouridine (FIAU), (E)-5-(2-bromovinyl)-2' deoxyuridine (BVdU), and 9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG or BW B759U) on the replication of Epstein-Barr virus (EBV) in vitro were evaluated and compared with that of acyclovir (ACV). The relative potencies of these drugs, on the basis of anti-EBV activity, were: FIAC = FIAU greater than FMAU greater than DHPG greater than BVdU greater than ACV; on the basis of the therapeutic index they were: BVdU greater than DHPG greater than FIAC greater than ACV greater than FIAU greater than FMAU. Differential inhibition of EBV-associated polypeptides by these drugs was observed. PMID- 2992369 TI - Comparison of the in vitro and in vivo antiherpes virus activities of the acyclic nucleosides, acyclovir (Zovirax) and 9-[(2-hydroxy-1 hydroxymethylethoxy)methyl]guanine (BWB759U). AB - The antiherpes virus activities of acyclovir and its close analogue 3-[(2-hydroxy 1-hydroxymethylethoxy)methyl]guanine (BWB759U) were compared in vitro and in vivo. The activities of both compounds against herpes simplex virus and varicella zoster virus were similar in the majority of cell lines. However, in mouse derived and HeLa cells, BSB759U was more effective than acyclovir against herpes simplex virus. Mutants of herpes simplex virus deficient in thymidine kinase and resistant to acyclovir were found to vary in their sensitivity to BWB759U. In two mouse models of herpes simplex virus infection BWB759U was more effective than acyclovir. PMID- 2992370 TI - Chemotherapy of Aujeszky's disease (pseudorabies) in the mouse by means of nucleoside analogues: bromovinyldeoxyuridine, acyclovir, and dihydroxypropoxymethylguanine. AB - Pseudorabies virus (PRV) infection was established in mice by means of inoculating the ear flap. The infection was universally fatal once clinical signs appeared. Bromovinyldeoxyuridine (BVDU) was a potent inhibitor of PRV in vitro, but this drug failed to protect mice and produced only marginal reductions in virus titre and slight prolongation of survival. Acyclovir (ACV) and dihydroxypropoxymethylguanine (DHPG) were both less active than BVDU when tested against the virus in BHK cells, yet DHPG therapy was extremely effective in mice; it reduced virus titres markedly and resulted in the long-term survival of mice given a potentially lethal infection. When ACV and DHPG were tested in vitro using murine rather than hamster cells, these compounds, especially DHPG, were shown to be much more active against PRV. PMID- 2992372 TI - Evidence for IFN-beta heterogeneity in a substrain of Namalwa cells. AB - A substrain of Namalwa cells, denoted substrain B, was grown in fermentors up to the 100-L scale, and was induced with Sendai virus to produce interferon (IFN). The titer of the crude IFN varied extensively between different batches; part of the variation was caused by a differential expression of IFN-alpha and IFN-beta. More than 80% of the IFN activity was IFN-beta by several criteria. A two-step purification procedure was developed and the resulting preparation had a specific activity of approximately 10(6) U/mg protein. The IFN-beta type was found to be heterogeneous, and could be separated into several components, which probably represented post-translational modifications of one molecule. PMID- 2992373 TI - pULB113, an RP4::mini-Mu plasmid, mediates chromosomal mobilization and R-prime formation in Erwinia amylovora, Erwinia chrysanthemi, and subspecies of Erwinia carotovora. AB - The RP4::mini-Mu plasmid pULB113, transferred from Escherichia coli strain MXR, was stable and transfer proficient in Erwinia amylovora strain EA303, E. carotovora subsp. atroseptica strain ECA12, E. carotovora subsp. carotovora strain ECC193, and E. chrysanthemi strain EC183. The plasmid mobilized an array of Erwinia sp. chromosomal markers (E. amylovora: his+,ilv+,rbs+,ser+,thr+;E. chrysanthemi:arg+,his+,ilv+,leu+; E. carotovora subsp. atroseptica: arg+,gua+,leu+,lys+,pur+,trp+; E. carotovora subsp. carotovora: arg+,gua+,leu+,lys+,out+[export of enzymes],pur+,trp+), suggesting random interactions of the plasmid with the chromosomes. In E. carotovora subsp. carotovora, pULB113-mediated two-factor crosses revealed linkage between three auxotrophic markers and the out loci. The export of pectate lyase, polygalacturonase, and cellulase and the maceration of potato tuber tissue occurred with Out+, but not Out-, strains of E. carotovora subsp. carotovora, indicating the importance of enzyme export in plant tissue maceration. Erwinia sp. donors harboring pULB113 complemented mutations in various biosynthetic and catabolic genes (arg, gal, his, leu, met, pro, pur, thy) in Escherichia coli recA strains. Escherichia coli transconjugants harbored pULB113 primes as indicated by the cotransfer of Erwinia genes and pULB113 markers and a change in plasmid mass. Moreover, the PstI and SmaI cleavage patterns of selected pULB113 primes were different from those of pULB113. pULB113 primes carried DNA insertions ranging from 3 to about 160 kilobases. These findings indicate that pULB113 is useful for in vivo gene cloning and genetic analysis of various enterobacterial phytopathogens. PMID- 2992371 TI - Preclinical assessment of topical treatments of herpes simplex virus infection: 5% (E)-5-(2-bromovinyl)-2'-deoxyuridine cream. AB - The potential efficacy of topical therapy with (E)-5-w-bromovinyl)-2' deoxyuridine (BVDU) for cutaneous herpesvirus infection was evaluated in vitro and in guinea pigs. Drug sensitivity testing against herpes simplex virus type 1 strain E115 revealed an ID50 of 0.008 microgram/ml for BVDU and 0.19 microgram/ml for acyclovir (ACV). In vitro drug diffusion studies showed poor penetration of guinea pig skin by BVDU from the cream compared to BVDU from dimethylsulfoxide (DMSO) (0.04 vs. 1.5 microgram/cm2 per h). 5% BVDU cream, 5% BVDU/DMSO, and 5% ACV in polyethylene glycol (PEG) were then compared in the treatment of experimental dorsal cutaneous HSV-1 infection in guinea pigs. Lesion number, total lesion area and virus titer were reduced by all three formulations compared to control sites treated with the corresponding drug vehicles (P less than or equal to 0.01). BVDU cream effected a greater reduction in lesion number (20+ vs. 13%) and total lesion area (40% vs. 28%) than did ACV/PEG and a significantly greater decrease in virus titer (990% vs. 55%, P less than 0.002). BVDU/DMSO was clinically twice as effective as BVDU cream (P less than or equal to 0.01) and reduced lesion virus titers to a similar degree. The results of these studies show that BVDU is a more potent virus-inhibitory agent than ACV in vitro and is superior to topical ACV in vivo when formulated in a simple aqueous cream. The marked efficacy of BVDU/DMSO in the animal model demonstrates the potential of this antiviral if drug delivery is improved. PMID- 2992374 TI - Inhibition of Clostridium botulinum 52A toxicity and protease activity by sodium acid pyrophosphate in media systems. AB - The effects of two pH levels (5.55 or 5.85) in combination with 0.4% sodium acid pyrophosphate (SAPP), NaH2PO4 X H2O, Na2HPO4 X 7H2O, or NaCl on the growth and toxicity of Clostridium botulinum 52A were studied. Absorbancy measurements at 630 nm, microscopic observations, and the mouse bioassay procedure were used to observe the effects. At pH 5.55 and 5.85 most control cultures exhibited toxicity when cell lysis began. Vegetative cell development was normal (4 micron long; 1 micron wide). SAPP-containing (0.4%) treatment cultures displayed similar growth and lysis but no or delayed (48 h) toxicity. Cells grown in the SAPP treatment culture were longer and wider (6 micron long; 1.5 micron wide) than in most other treatment cultures. Trypsinization of nontoxic supernatants from 0.4% SAPP resulted in toxicity. Addition of 0.4% SAPP to toxic C. botulinum supernatant delayed but did not prevent death of mice. The addition of various levels of SAPP to toxic supernatants resulted in a decrease in zone size with an increase in the level of SAPP (9 mm with 0.4% SAPP to 7 mm with 1.0% SAPP), using a dual substrate protease assay. A decrease in the zone size also occurred with the supernatant from cultures grown in the presence of SAPP and with Bacillus polymyxa protease dilutions containing 0.4% SAPP. Results suggest that the actual production or function of the protease responsible for toxin activation may have been inhibited by the presence of SAPP. PMID- 2992375 TI - Construction of a cosmid clone library of Pseudomonas syringae pv. phaseolicola and isolation of genes by functional complementation. AB - A genomic library constructed from a wild-type strain of Pseudomonas syringae pv. phaseolicola in the broad-host-range cosmid vector pVK102 was used to isolate wild-type genes by complementation of Tn5-induced auxotrophic mutants. Selection pressure was required for maintenance of the vector and members of the library in strains of P. syringae. PMID- 2992376 TI - Evidence for plasmid- and chromosome-borne multiple nif genes in Rhizobium fredii. AB - Rhizobium fredii is a fast-growing rhizobium isolated from the primitive Chinese soybean cultivar Peking and from the wild soybean Glycine soja. This rhizobium harbors nif genes on 150- to 200-megadalton plasmids. By passage on acridine orange plates, we obtained a mutant of R. fredii USDA 206 cured of the 197 megadalton plasmid (USDA 206C) which carries both nif and nod genes. This strain, however, has retained its symbiotic effectiveness. Probing EcoRI digests of wild type and cured plasmid DNA with a 2.2-kilobase nif DH fragment from Rhizobium meliloti has shown four homologous fragments in the wild-type strain (4.2, 4.9, 10, and 11 kilobases) and two fragments in the cured strain (4.2 and 10 kilobases). EcoRI digests of total DNA show four major bands of homology (4.2, 4.9, 5.8, and 13 kilobases) in both the wild-type and cured strains. The presence of major bands of homology in the total DNA not present in the plasmid DNA indicated chromosomal nif genes. Probing of HindIII digests of total and plasmid DNA led to the same conclusion. Hybridization to the smaller plasmids of USDA 206 and USDA 206C showed the presence of nif genes on at least one of these plasmids, explaining the nif homology in the USDA 206C plasmid digests. PMID- 2992378 TI - Inactivation of glutathione peroxidase by superoxide radical. AB - The selenium-containing glutathione peroxidase, when in its active reduced form, was inactivated during exposure to the xanthine oxidase reaction. Superoxide dismutase completely prevented this inactivation, whereas catalase, hydroxyl radical scavengers, or chelators did not, indicating that O2 was the responsible agent. Conversion of GSH peroxidase to its oxidized form, by exposure to hydroperoxides, rendered it insensitive toward O2. The oxidized enzyme regained susceptibility toward inactivation by O2 when reduced with GSH. The inactivation by O2 could be reversed by GSH; however, sequential exposure to O2 and then hydroperoxides caused irreversible inactivation. Reactivity toward CN- has been used as a measure of the oxidized form of GSH peroxidase, whereas reactivity toward iodoacetate has been taken as an indicator of the reduced form. By these criteria both O2 and hydroperoxides convert the reduced form to oxidized forms. A mechanism involving oxidation of the selenocysteine residue at the active site has been proposed to account for these observations. PMID- 2992377 TI - Cloning and expression of a Streptococcus cremoris proteinase in Bacillus subtilis and Streptococcus lactis. AB - Previously, curing experiments suggested that plasmid pWV05 (17.5 megadaltons [Md]) of Streptococcus cremoris Wg2 specifies proteolytic activity. A restriction enzyme map of pWV05 was constructed, the entire plasmid was subcloned in Escherichia coli with plasmids pBR329 and pACYC184. A 4.3-Md HindIII fragment could not be cloned in an uninterrupted way in E. coli but could be cloned in two parts. Both fragments showed homology with the 9-Md proteinase plasmid of S. cremoris HP. The 4.3-Md HindIII fragment was successfully cloned in Bacillus subtilis on plasmid pGKV2 (3.1 Md). Crossed immunoelectrophoresis of extracts of B. subtilis carrying the recombinant plasmid (pGKV500; 7.4 Md) showed that the fragment specifies two proteins of the proteolytic system of S. cremoris Wg2. PGKV500 was introduced in a proteinase-deficient Streptococcus lactis strain via protoplast transformation. Both proteins were also present in cell-free extracts of S. lactis(pGKV500). In S. lactis, pGKV500 enables the cells to grow normally in milk with rapid acid production, indicating that the 4.3-Md HindIII fragment of plasmid pWV05 specifies the proteolytic activity of S. cremoris Wg2. PMID- 2992379 TI - Adenosinetriphosphate sulfurylase from Penicillium chrysogenum: steady-state kinetics of the forward and reverse reactions, alternative substrate kinetics, and equilibrium binding studies. AB - The kinetics of the forward ATP sulfurylase-catalyzed reaction were examined using a new assay based on 32PPi released from [gamma-32P]MgATP in the presence of inorganic sulfate. Replots yielded Vmaxf = 6.6 units mg protein-1, KmA = 0.13 mM, Kia = 0.33 mM, and KmB = 0.55 mM, where A = MgATP and B = SO2-4. Thiosulfate, a dead-end inhibitor of the reaction, was competitive with sulfate and noncompetitive with respect to MgATP. The ratio kcat/KmA was determined for several alternative inorganic substrates, B, where A = MgATP and B = SO2-4, SeO2 4, MoO2-4, WO2-4, or CrO2-4. For SO2-4 and SeO2-4, the ratio was 5-6.5 X 10(4) M 1 S-1; for the others, the ratio was 5.8-7.3 X 10(5) M-1 S-1. The results support a random addition of MgATP and inorganic substrate. The kinetics of the reverse reaction were examined using a new assay based on 35SO2-4 release from [35S]APS (adenosine 5'-phosphosulfate) in the presence of MgPPi. Reciprocal plots were linear, intersecting below the horizontal axis. Replots yielded Vmaxr = 50 units mg protein-1, KmQ = 0.3 microM, Kiq = 0.04 microM, and KmP = 4 microM, where Q = APS and P = PPi (total of all species). MgATP and SO2-4 were both competitive with APS and noncompetitive with respect to MgPPi. Taken together with earlier results suggesting that APS is competitive with both MgATP and SO2-4 and that MgPPi is noncompetitive with respect to both substrates, the qualitative results point to a random A-B, ordered P-Q kinetic mechanism. The Scatchard plot for [35S]APS binding was curved, indicating either negative cooperativity or more than a single class of sites. [gamma-32P]MgATP displayed half-site saturation in the presence of saturating FSO-3. PMID- 2992380 TI - Purification and characterization of an abscisic acid-inducible anionic peroxidase associated with suberization in potato (Solanum tuberosum). AB - An anionic peroxidase (EC 1.11.1.7), thought to be involved in suberization, was purified 110-fold from wound-healing slices of Solanum tuberosum by a combination of ammonium sulfate fractionation, Sephadex G-100 gel filtration, isoelectric focusing, and phenyl-Sepharose CL-4B chromatography in 24% yield. The purified enzyme was homogeneous as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and horizontal thin-layer polyacrylamide gel electrophoresis. The molecular weight of the enzyme was estimated to be 47,000 by both Sephadex G-100 gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This peroxidase was found to be a glycoprotein containing about 17% carbohydrate, approximately one-quarter of which was shown to be glucosamine residues. It was found to have an isoelectric point of 3.15. An anionic peroxidase was also isolated from abscisic acid-treated callus tissue culture of S. tuberosum by the above purification procedure. The two enzymes were shown to be immunologically similar, if not identical, based on their cross-reactivity with rabbit antibody prepared against the peroxidase from wound-healing slices, whereas the major cationic peroxidase from wound-healing slices did not cross-react with this antibody. The anionic enzyme from both sources showed very similar specific activities when assayed with a range of substrates, whereas the specific activities found for the cationic isozyme isolated from wound-healing slices were quite different. PMID- 2992381 TI - 1H NMR spectroscopy of cytochrome cd1 derivatives. AB - Proton nuclear magnetic resonance spectra are reported for cytochrome cd1 from Pseudomonas aeruginosa (ATCC 19429) in several forms including complexes of the ferricytochrome with cyanide, azide, and fluoride, a quasi-apo form in which the noncovalently associated heme d1 has been removed but the covalently bound heme c is retained, and the reduced state of both native and the quasi-apo forms. Comparisons are made to the previously reported spectrum of ferricytochrome cd1. The following points are made. The spectra of the azide and fluoride complexes and the ferric quasi-apo form show perturbation of resonances assignable to the site of heme d1, and leave relatively unperturbed resonances assignable to the site of heme c. The heme d1 associated resonances are at 46.0, 35.4, 23.3, 17.5, 2.9, and 16 ppm, and the heme c associated resonances are at 42.0, 33.7, 15.0, 13.9, -7.5, -14, and -33 ppm in native ferricytochrome cd1. The similarity of the hyperfine resonances of the ferric quasi-apo from to the heme c resonances of intact ferricytochrome cd1 is evidence that removal of heme d1 leaves the heme c binding site relatively unaltered. Linewidths and relaxation times suggest that the relaxation times of the unpaired electron spins of the ferric hemes c and d1 are on the same order of magnitude. Although it is paramagnetic, ferrocytochrome cd1 does not demonstrate an experimentally detectable hyperfine shifted spectrum under present conditions. Possible reasons for this are discussed. The presence of a narrow resonance at -2.8 ppm in both ferrocytochrome cd1 and the reduced state of the quasi-apo form suggests that methionine may be a ligand to heme c. PMID- 2992382 TI - Structure-function relationships of S-carboxymethyl methionine27 glucagon. AB - Carboxymethylation of glucagon and subsequent purification of the hormone has provided a derivative modified by the addition of bulk to the methionine at position 27 without a net charge alteration in the side chain. Unreacted glucagon was removed after methylation of the methionine which provides a positively charged chromatographic handle. The derivative has a half-maximum concentration for binding of 5.3 nM and is a full agonist. These findings along with those provided by methylation of the methionine indicate that a positive charge rather than bulk on the methionine side chain disrupts the binding of hormone to its receptor. The S-carboxymethyl derivative lacks the concentration-dependent aggregation characteristic of glucagon at pH 10.2 as does the S-methyl derivative but increases its helical content in 30% 2-chloroethanol to the same extent as native and S-methyl hormone. Full activity of the S-carboxymethyl methionine27 glucagon does not favor the existence of the globular structure proposed by Korn and Ottensmeyer [(1983) J. Theor. Biol. 105, 403] as the binding species whereas multiple considerations do favor a flexible hormone with nucleation followed by conformational changes for complete binding and activation. Isotopic enrichment using labeled iodoacetate is feasible and can provide more definitive structural information. PMID- 2992384 TI - On the mechanism of glycolysis stimulation by neutral detergents in 3T3 and Ehrlich ascites tumor cells. AB - Glycolysis of 3T3 and Ehrlich ascites tumor cells was greatly enhanced by Nonidet P-40 or Triton X-100 at about 100 micrograms/mg cell protein. This enhanced glycolysis was partly sensitive to rutamycin and partly sensitive to ouabain, suggesting that the detergent released the control of the ATPase of the mitochondria and of the plasma membrane Na+K+-ATPase. Nonidet P-40 had no effect on glycolysis in cell-free extracts from Ehrlich ascites tumor cells to which soluble mitochondrial ATPase was added. Measuring ouabain-sensitive 22Na efflux and using ouabain-sensitive lactate production as a measure of ATP hydrolysis by the Na+K+ pump, it was shown that Nonidet P-40 greatly decreased the efficiency of the Na+K+ pump. Quercetin increased the efficiency of pumping in EAT cells both in the absence and presence of the detergent. PMID- 2992383 TI - Comparison of the spectroscopic and saccharide binding properties of lentil and pea isolectins. AB - The lentil isolectins, CMLcH A and CMLcH B, and pea isolectins, CMPSA A and CMPSA B, are compared in terms of their spectroscopic and saccharide binding properties. The paramagnetic contribution to the solvent proton magnetic relaxation dispersion profiles of solutions of the isolectins of each protein are found to be essentially identical. Electron paramagnetic resonance spectra suggest a high degree of octahedral symmetry at the Mn2+ site for both pairs of isolectins. The near-ultraviolet absorption spectra of CMLcH A and CMLcH B are identical, as are the spectra of CMPSA A and CMPSA B. Carbohydrate binding activities of the isolectins of each protein are compared using hemagglutination, precipitation, and precipitation-inhibition assays, and are found to be identical, although the activities of CMLcH and CMPSA differ somewhat. These results demonstrate that the spectroscopic and saccharide binding properties of the isolectins of CMLcH are essentially identical, as are those of the isolectins of CMPSA, and suggest that native mixtures of the isolectins may be treated as single proteins in further studies. PMID- 2992385 TI - A comparison of the cyclic nucleotide-dependent protein kinases using chemical cleavage at tryptophan and cysteine. AB - cGMP-dependent protein kinase (G-kinase) and the regulatory subunit of type I (RI) cAMP-dependent protein kinase (A-kinase) both contain a phosphorylation site located near the NH2 terminus of each enzyme. These sites can be utilized as convenient markers for the determination of the position of an amino acid residue susceptible to either chemical or enzymatic digestion. Using the tryptophan specific reagent, N-chlorosuccinimide, the approximate location along the polypeptide chain of six reactive tryptophans in G-kinase and three reactive residues in RI were identified. Similarly, cleavage with cyanide was used to locate free and disulfide-bonded cysteines in both proteins. The approximate positions of nine cysteines in G-kinase were determined along with the location of the interchain disulfide bond and an intrachain disulfide bond. RI was found to contain three cyanide-reactive cysteines, two of which are involved in interchain disulfide bonding. A comparison of the positions of the cysteines and tryptophans determined by chemical cleavage in G-kinase and RI, with the positions of cysteine and tryptophan in the known sequence of the type II A kinase, support the structural relationships between these enzymes. Comparison with subsequently reported primary sequences of all three enzymes indicates the limits of precision of this chemical cleavage procedure. PMID- 2992386 TI - Functional characterization of the mitochondrial cytochrome b-c1 complex: steady state kinetics of the monomeric and dimeric forms. AB - The QH2:cytochrome c oxidoreductase activity of the isolated bovine heart cytochrome b-c1 complex resolved into monomeric and dimeric form was titrated with three different inhibitors of electron transfer, antimycin, myxothiazol, and 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole (UHDBT). In all cases one inhibitor molecule per cytochrome c1 was found necessary to block completely the activity of both molecular forms of the enzyme. The antimycin-sensitive cytochrome c reduction catalyzed by the b-c1 complex was also studied as a function of increasing concentrations of either cytochrome c or quinol. Double-reciprocal plots of the activity of the monomeric enzyme were found linear either when the concentration of cytochrome c or of quinol derivatives, 2,3-dimetoxy-5-methyl-6 decyl-1,4-benzoquinol (DBH), and 2-methyl-3-undecyl-1,4-naphthoquinol (UNH), was changed. Cytochrome c reductase activity of the dimeric b-c1 complex also showed a linear Lineweaver-Burk plot as a function of cytochrome c concentrations. In contrast to the monomeric enzyme, however, dimers of the b-c1 complex express a clear nonlinear kinetic behavior toward quinol derivatives, with two apparent Km values differing approximately by one order of magnitude (about 3-4 and about 20 30 microM). At saturating quinol concentrations the activity of the dimeric enzyme becomes two to three times higher than that of monomers. The nonlinear kinetic plots were found to be the same at different temperatures and different cytochrome c concentrations. The data suggest that although the monomer of the b c1 complex appears to be the functional unit of the enzyme, the dimer is more active. A regulatory role of the dimerization process resulting in an increase of the electrons flux through the enzyme is postulated. PMID- 2992387 TI - The relationship of alkaline phosphatase, CaATPase, and phytase. AB - Alkaline phosphatase, highly purified from bovine intestinal mucosa, has significant hydrolytic activity against phytate and CaATP. Phytase and CaATPase activities require quite different assay conditions than those which are optimal for conventional alkaline phosphatase substrates such as 4-nitrophenyl phosphate. We have used affinity chromatography and antibody recognition to demonstrate that the phytase and CaATPase activities are not due to contaminating enzymes, but are intrinsic activities of intestinal alkaline phosphatase. All of the phytase and CaATPase activities present in crude extracts of bovine intestinal mucosa can be accounted for by alkaline phosphatase. Apparently neither phytase nor CaATPase exist in this tissue as independent enzymes. Specific substrates which require assay conditions quite different from the conventional 4-nitrophenyl phosphate substrate may account for the physiological function of "alkaline phosphatase." PMID- 2992388 TI - The effects of "oxygen radicals" generated in the medium on lenses in organ culture: inhibition of damage by chelated iron. AB - Rat lenses in organ culture were exposed to activated species of oxygen generated in the culture medium either by xanthine oxidase and hypoxanthine or by riboflavin and visible light, two systems which have been shown to produce superoxide and H2O2. In each case there was marked damage to carrier-mediated transport systems of the lens. Under standard culture conditions this damage was strongly inhibited by catalase, but not by superoxide dismutase (SOD). By the addition to the medium of chelated iron, hydroxyl radicals were produced in a Fenton reaction with a concomitant decrease in H2O2 levels. With both oxygen radical-generating systems, the addition of chelated iron strongly inhibited lens damage. This inhibitory effect could be reversed by the addition of SOD with the chelated iron. Under such conditions SOD converts superoxide anion to H2O2, thereby preventing reduction of the chelated iron and thus stopping the generation of hydroxyl radicals. Increased lens damage following addition of SOD to the iron-containing systems correlated with higher H2O2 concentrations, and was inhibited by catalase. These findings suggest that, when generated in the fluids surrounding the lens, H2O2 poses a much greater oxidative stress for the lens than do the superoxide or hydroxyl free radicals. PMID- 2992390 TI - A protein inhibitor of calmodulin-regulated cyclic nucleotide phosphodiesterase in amphibian ovaries. AB - The cytosol fraction of an extract of Xenopus laevis ovaries contains a protein inhibitor that can specifically block the activation of calmodulin-sensitive cyclic nucleotide phosphodiesterase (PDE I) found in that tissue. This inhibitor was purified by DEAE-cellulose chromatography, gel filtration on Sephacryl S-200, and affinity chromatography on calmodulin-Sepharose. It has a molecular weight of approximately 90,000, and is heat-labile and susceptible to inactivation by chymotrypsin. The inhibitor blocks calmodulin activation of cyclic nucleotide phosphodiesterases from amphibian ovary and bovine brain and of the myosin light chain kinase from rabbit smooth muscle, but does not affect the activity of a calmodulin-insensitive cyclic nucleotide phosphodiesterase. The inhibitor not only affects the activation of Xenopus PDE I and of the bovine brain phosphodiesterase by calmodulin, but also inhibits the stimulation of these enzymes by lysophosphatidylcholine. The inhibitor also acts on PDE I activated by partial tryptic proteolysis, but the enzyme fully activated by trypsin is only slightly susceptible to inhibition by this protein. The inhibition of PDE I activation caused by this ovarian factor can be reversed by adding excess amounts of calmodulin or lysophosphatidylcholine. The presence of this inhibitor provides a possible explanation for the previously observed inactivity of PDE I in vivo. PMID- 2992389 TI - Characterization of cucumber ascorbate oxidase and its reaction with hexacyanoferrate (II). AB - The spectroscopic features of cucumber ascorbate oxidase (AOase) and its type-2 copper-depleted (T2D) derivative, and the electron pathway among the copper sites in the enzyme have been investigated. The electronic and CD spectra of native and T2D AOase in the visible region bear a striking resemblance to those of plastocyanin or azurin, which contain type-1 copper alone. The electronic absorption shoulder of the native enzyme at around 330 nm for the native enzyme which has been assigned to type-3 copper disappears with the depletion of the type-2 copper. The reduction of AOase with a large excess of hexacyanoferrate(II) results in a selective reduction of the type-2 Cu, giving rise to an additional EPR-detectable species which is considered to be originated from partly reduced type-3 copper. The type-1 copper is, however, not reduced even in the presence of excess hexacyanoferrate(II). The redox potential of type-1 Cu was determined to be +350 mV, which is distinctly lower than that of hexacyanoferrate(II-III). Type 2 copper was supposed to be a mediator of the electron transfer between type-1 and type-3 coppers in consideration of the extremely low activity of the T2D enzyme under the same condition. A comparison of the electron pathway in AOase with that in laccase is also argued. PMID- 2992391 TI - Identification and partial characterization of bovine heart cytosolic phosphorylase phosphatases. AB - The properties of phosphatases in bovine heart cytosol were studied. Two isozymic forms of protein phosphatase H (H-1 and H-2) were resolved by chromatography on DEAE-Sephacel. The two isoenzymes had identical physical properties (Mr 260,000, 7.9 S). Treatment with 80% ethanol activated both isozymes and converted H-1 to a Mr 35,500 form and H-2 to Mr 67,000 and Mr 35,500 forms. Both H-1 and H-2 and their lower Mr activated forms had essentially identical Km values for phosphorylase a. The heart cytosol also contained a latent phosphatase (Fc) which could be activated by preincubation with either ATP X Mg and an activating factor (FA), or by Mn/trypsin treatment. The latter procedure converted the latent Fc (Mr 200,000) to a Mn2+-independent Mr 34,500 form. Both activated forms of Fc had similar Km values which were fourfold lower than the affinity of the protein phosphatase H forms for the phosphorylase a substrate. PMID- 2992392 TI - Coregulation of collagenase and collagenase inhibitor production by phorbol myristate acetate in human skin fibroblasts. AB - Phorbol myristate acetate (PMA), a tumor promotor known to stimulate collagenase production in fibroblasts and endothelial cells, was examined with regard to its ability to regulate the expression of the collagenase inhibitor secreted by human skin fibroblasts. Confluent human skin fibroblasts were incubated with concentrations of PMA ranging from 10(-11) to 10(-7) M, and the conditioned medium was analyzed by enzyme-linked immunosorbent assay for both immunoreactive collagenase and collagenase inhibitor. PMA stimulated the production of both collagenase and collagenase inhibitor in several cell lines to maximal rates that were very similar, 300 to 350 vs 230 to 330 pmol 10 micrograms DNA-1 48 h-1, respectively. Due to differences in the basal levels of expression of these proteins, such rates reflected a two- to sevenfold stimulation in collagenase production, in comparison to a more uniform two- to threefold enhancement in inhibitor synthesis. Production of inhibitor was 50% of maximal at 7 X 10(-9) M and maximal at 10(-7) M phorbol. This concentration-dependent effect was very similar to that observed for collagenase expression. Total protein synthesis by the phorbol-conditioned cells, as studied by incorporation of [3H]leucine into newly synthesized protein, was not significantly increased, nor was cellular DNA content. The onset of the effect of PMA on inhibitor production occurred between 4 and 8 h, was maximal by 8 h, and continued undiminished for at least another 64 h. After the first 8 h, inhibitor production continued at a roughly constant rate of approximately 10 pmol 10 micrograms DNA-1 h-1. Interestingly, following the removal of phorbol from culture medium, such fibroblasts continued to produce increased quantities of inhibitor protein for at least 72 h. Metabolic labeling studies in which fibroblasts were exposed to [3H]leucine followed by immunoprecipitation using inhibitor-specific antibody suggested that stimulation of inhibitor production by PMA was mediated via an increased synthesis of new inhibitor protein. Therefore, in response to the tumor promoter, PMA collagenase and collagenase inhibitor expression by human skin fibroblasts appear to be coregulated. PMID- 2992394 TI - Oxidation and reduction of cytochrome c bound to the phosphoprotein phosvitin. AB - The oxidation-reduction reactions and structural characteristics of phosvitin bound cytochrome c were examined at various ratios of cytochrome c to phosvitin. At binding ratios below half the maximum, the rate constants for the oxidation reactions with cytochrome c oxidase and ferricyanide and the rate constants for the reduction reactions with cytochrome b2 and ascorbate were low, but at higher ratios these rate constants gradually increased to that of free cytochrome c and, in particular, the rate constant for oxidation by cytochrome c oxidase was raised to two to three times that of the free form. This binding-ratio dependence of the rate constants for the oxidation and reduction reactions was different from that of the net charge of the cytochrome c-phosvitin complex, implying that the negative charges of phosvitin are unlikely to modulate the rates. In contrast, the broadening of the NMR signals for the heme and methionine-80 methyl groups and the conformational transition in the vicinity of the heme moiety on change from the native to the cyanide-bound or urea-denatured form of cytochrome c showed a similar binding-ratio dependence to the rate constants for the oxidation and reduction reactions. Since the conformation and electronic structure in the heme environment of ferric and ferrous cytochromes c were not changed significantly by binding to phosvitin, and since the binding strength of cytochrome c to phosvitin at binding ratios below half the maximum is different from that at higher ratios, these findings suggest that a difference in the movement of cytochrome c in its complex with phosvitin may modulate its oxidation reduction reactions. PMID- 2992393 TI - A cytosolic flavin-containing enzyme catalyzing reduction of cytochrome c in Trypanosoma cruzi: kinetic studies with cytochrome c as substrate. AB - The kinetic mechanism of cytochrome c reduction by a Trypanosoma cruzi cytosolic flavoenzyme was investigated by initial velocity determinations, by product inhibition patterns, and by the characteristics of inhibition by analogs. The data suggest a two-site ping-pong mechanism in which NADPH reduces the flavin, which is then reoxidized in two one-electron steps by reaction with two molecules of cytochrome c. The two-site nature of the mechanism is probably related to the dimeric nature of the enzyme, and the binding sites of cytochrome c and NADPH are probably on opposite sites of the FAD. PMID- 2992395 TI - [Update of human tumor clonogenic assay in carcinoma of the lung]. AB - The human tumor clonogenic assay (HTCA) is a double-layer agar technique, which provides an in vitro prediction of the response of an individual patient's tumor to an antitumor agent. This paper briefly provides an outline of HTCA and its potential use in chemotherapy on patients with lung cancer. In our experience with culturing 123 carcinomas of the lung, 105 specimens (85%) could be subject to more than 5 chemosensitivity tests each by modifying the preparation method of single cell suspension in this system. Growth rate was improved in all types of primary human lung cancer with reasonable consistency. Further, metastatic tumors were capable of being successfully grown in a high percentage of cases, which was comparable to the results obtained for other kinds of tumors. There was no correlation of growth or cloning efficiency with histology, source, or previous chemotherapy. Using 50% or more inhibition on to colony formation as the criterion for chemosensitivity, response rates to vindesine or mitomycin C were 19% or 16%, respectively. The in vitro response rates of these or almost all other antitumor drugs seemed to be comparable to the clinical responses reported by various investigators. Correlation between in vitro chemosensitivity in HTCA and clinical response has been evaluated by various investigators, and the pooled data have demonstrated a good association between in vitro drug sensitivity and clinical response or lack of response. In lung cancer, HTCA had a 57% true positive rate and an 85% true negative rate for the prediction of drug sensitivity and resistance, respectively, of cancer patients to specific chemotherapeutic drugs. Although the system still has to undergo modification to resolve a number of theoretical and practical problems, it has potential uses in lung cancer chemotherapy. PMID- 2992396 TI - A retrospective cohort mortality study of a phosphate fertilizer production facility. AB - A retrospective cohort mortality study of phosphate fertilizer production workers was undertaken to determine whether this group is at increased risk of dying from any cause, particularly from lung cancer. A total of 3,199 workers who had ever been employed at one facility were included in this investigation. These workers were followed for vital status ascertainment from their first date of employment up to December 1, 1977, or the date of death, whichever occurred first. Overall, no statistically significant elevations in cause-specific mortality were observed for the entire study population. However, when the analysis was stratified by duration of employment, and length of follow-up, a statistically significant (P less than .05) excess in lung cancer mortality was observed among workers with more than 10 yr of employment and follow-up (standardized mortality ratio = 411). Because of the small number of deaths involved, and because we had prior knowledge of a lung cancer cluster at this plant, we believe that these findings should be viewed as suggestive, and that other investigations in plants with similar exposures are needed to clarify whether an occupationally related lung cancer excess truly exists. PMID- 2992397 TI - Exclusion of the alpha 1(II) collagen structural gene as the mutant locus in type II Ehlers-Danlos syndrome. AB - We have used a high frequency site polymorphism within the human pro-alpha 1(II) collagen gene (COL2A1) in order to examine the segregation of this gene within a large pedigree with type II Ehlers-Danlos syndrome (EDS). The EDS gene and the collagen gene segregate independently within the pedigree and therefore COL2A1 can be excluded as the mutant locus. PMID- 2992399 TI - Acute monoarthritis associated with lipid liquid crystals. AB - We describe three women with unexplained acute monoarthritis and birefringent lipid spherules or lipid liquid crystals in their synovial fluid. Synovial fluid aspirated within 24 hours of onset showed leucocyte counts ranging from 10 X 10(9) to 46 X 10(9)/1 with 91-95% polymorphonuclear cells. Numerous positively birefringent 2-6 micron microspherules were seen inside and outside polymorphonuclear cells. These were dissolved in alcohol-ether but were not digested by uricase. Other non-birefringent globules of similar size that stained with Sudan black B were also seen. Transmission electron microscopy identified osmiophilic homogeneous or multilamellated material in phagocytic vacuoles. Serum lipids, lipase, and repeated joint radiographs were normal. Arthritis subsided completely after treatment with colchicine in one patient and nonsteroidal anti inflammatory agents in two. Thus birefringent microspherules can be found in association with acute, otherwise unexplained, arthritis. It seems possible that certain lipid liquid crystals may induce synovial inflammation similar to inflammation seen in other crystal induced arthritides. PMID- 2992398 TI - Abnormal responses of rheumatoid arthritis lymphocytes to Epstein-Barr virus infection in vitro: evidence for multiple defects. AB - Blood lymphocytes from 53 patients with rheumatoid arthritis (RA) and 44 controls were cultured with the polyclonal B cell activator Epstein-Barr virus (EBV). Culture supernatants were removed at weekly intervals and the amount of IgM secreted by the lymphocytes measured by an enzyme-linked immunosorbent assay (ELISA). Three major differences in the pattern of EBV-induced IgM synthesis by RA versus control lymphocytes were observed. Lymphocytes from RA patients, in general, produced less IgM after one week in culture than controls. In contrast, they increased their IgM secretion significantly by the end of the second week, whereas control lymphocyte cultures showed little change in IgM secretion at this time. Control lymphocytes from EBV seropositive individuals produced undetectable amounts of IgM after five weeks in culture. However, lymphocytes from 40% of the RA patients, even though they were EBV seropositive, secreted greater than 2000 ng/ml (microgram/l) IgM after five weeks. The data are discussed in terms of defective B and T cell responses to EBV in lymphocytes from patients with RA. PMID- 2992400 TI - Bronchial adenoma: a review of 51 patients. AB - The cases of 51 patients with bronchial adenomas were reviewed. There were 43 bronchial carcinoids, 5 adenoid cystic carcinomas, 2 mixed tumors, and 1 mucoepidermoid carcinoma. The carcinoid group was divided into typical (31, 72%) and atypical (12, 28%) subgroups. Nine carcinoids (20%) were categorized as metastasizing adenomas; in this group, 7 lesions were atypical and 2 were typical. Thirty-two lobectomies, 7 bilobectomies, 8 pneumonectomies, 2 sleeve resections, and 2 tracheal resections were performed. Ten-year survival was 88% for patients with typical carcinoids and 59% for those with atypical carcinoids. In the group with adenoid cystic carcinoma, 1 patient died postoperatively, 1 had recurrence of the tumor, 2 were alive and free from disease 16 and 23 years later, and 1 died of heart disease at 11 years. The patient with mucoepidermoid carcinoma was alive without recurrence 15 years after operation. In conclusion, bronchial adenomas of the carcinoid type are potentially malignant. Their prognosis depends on the histology of the tumor, and on the presence of metastasis to the regional lymph nodes and distant organs. PMID- 2992401 TI - Similarities and differences in the effects of verapamil, diltiazem and nifedipine on arterial baroreflexes of anesthetized cats. AB - The effects of verapamil, diltiazem and nifedipine on baroreflex sensitivity (BS) were investigated in anesthetized cats at equihypotensive doses of 100, 100 and 10 micrograms/kg, respectively. The gain in BS (ms/mmHg) was calculated from the ratio delta heart period/delta systolic pressure following evoked rises and falls in arterial pressure. Verapamil and diltiazem significantly augmented BS while nifedipine depressed BS during evoked rises in arterial pressure. By contrast, BS was significantly depressed by diltiazem and nifedipine in response to depressor challenges with nitroprusside, while it was unaffected by verapamil. The augmentatory action of verapamil on BS after pressor challenges, was abolished by bilateral vagotomy while the inhibitory action of nifedipine was still evident after vagotomy, suggesting a vagal augmentatory action for verapamil and an involvement of the cardiac sympathetic nerves in the inhibitory action of nifedipine on BS. The differences between the actions of the 3 drugs on BS are not readily explainable by our data but they cannot be attributed to alterations of cardiac autonomic function since at the doses employed none of the drugs affected the cardiac responses to carotid occlusion, i.v. isoproterenol or electrical stimulation of the right vagus. It is concluded that BS is affected differently by the 3 chemically unrelated calcium channel blockers in a way which could either be contributory or deleterious to the antihypertensive effectiveness of these drugs. PMID- 2992402 TI - Identification and characterization of 3H-prazosin binding to alpha 1 adrenoceptors in bovine teat muscles. AB - The highly specific alpha 1-adrenoceptor antagonist 3H-prazosin was used to characterize alpha 1-adrenoceptors in smooth muscles of the cistern wall of teats of lactating cows. Binding was rapid, readily reversible, stereospecific and saturable. Scatchard analysis of saturation binding data on bovine teat membranes revealed that 3H-prazosin bound to a single class (Bmax = 128 +/- 13 fmol/mg of membrane protein) of noncooperative sites (Hill coefficient = 0.95 +/- 0.02) with high affinity (KD = 0.47 +/- 0.05 nM). Competition binding studies with 9 antagonists and 8 agonists, indicated that 3H-prazosin labeled the alpha 1 receptor. PMID- 2992403 TI - Clonidine-induced desensitization differentiates presynaptic alpha-2 adrenoceptors of the guinea-pig ileum and rat vas deferens. AB - Clonidine-induced inhibitions of twitch responses of coaxially stimulated guinea pig ileum and transmurally stimulated rat vas deferens were antagonized by yohimbine in a competitive manner. In contrast to rat vas deferens, clonidine caused desensitization of presynaptic alpha 2-adrenoceptors on cholinergic neurons of guinea-pig ileum. Cross desensitization was observed between clonidine and oxymetazoline. No cross desensitization between clonidine and morphine was observed and indicates that presynaptic alpha 2-adrenoceptors are involved in clonidine-induced desensitization of the presynaptic site in the guinea-pig ileum. PMID- 2992404 TI - Effects of oxamniquine on neuromuscular transmission. AB - The effects of oxamniquine on neuromuscular transmission have been investigated in some experimental animals in an effort to unveil some of the unwanted effects of this commonly prescribed schistosomicide. Results obtained show that relatively low concentrations of the drug (oxamniquine, 10-100 micrograms/ml) potentiated acetylcholine-, carbachol- and nicotine-evoked contractions of the toad (Bufo regularis) isolated rectus abdominis muscle. Moderate to high concentrations of the compound (oxamniquine, 500 micrograms/ml and above) contracted rectus abdominis muscle preparations, and depressed indirect electrically-induced twitches of the rat isolated hemidiaphragm and chick isolated biventer muscle preparations. The depression of electrically-evoked twitches of the chick biventer and rat phrenic nerve-hemidiaphragm muscle preparations in vitro induced by moderate to high concentrations of oxamniquine (500 micrograms/ml and above) were resistant to bath-applied physostigmine (1-15 micrograms/ml), but deepened by d-tubocurarine (3-10 micrograms/ml). These findings suggest that the neuromuscular blocking action of oxamniquine is probably post-junctional in origin. PMID- 2992405 TI - Effect of barium ion on p-aminohippurate transport in basolateral membrane vesicles isolated from rat kidney cortex. AB - To clarify the cause of the stimulation of p-aminohippurate (PAH) accumulation in rat kidney cortical slices by barium, an experiment was carried out with basolateral membrane vesicles isolated from rat kidney cortex. The effect of barium on PAH uptake by the membrane vesicles was compared with that of verapamil which also stimulated PAH accumulation in the slices. The enzyme marker for basolateral membrane, (Na+ + K+)- ATPase, was enriched 15-fold and the brushborder enzyme marker, alkaline phosphatase, was 1.3-fold in our membrane preparation. Contamination in this preparation by lysosomes, mitochondria and cytosol was also low but that by endoplasmic reticulum was slightly high as judged by the enzyme markers. PAH uptake by the membrane vesicles possessed the usual characteristics, i.e., sodium-dependence and probenecid-sensitivity. PAH uptake by the membrane vesicles was enhanced by barium, but not by verapamil. On the other hand, barium did not affect tetraethylammonium (TEA) uptake by the vesicles, and verapamil strongly inhibited it. Manganese also stimulated PAH uptake to the same extent as did barium, but calcium and strontium did not affect the uptake. Barium did not act on sodium transport in the membrane vesicles. An 'anion-sensitively transported lipophilic cation', triphenylmethylphosphonium iodide (TPMP), uptake was depressed by barium. These results suggest that barium stimulates selectively PAH uptake in basolateral membrane vesicles. Its stimulatory action may contribute at least partly to an increase in PAH accumulation in rat kidney cortical slices by this ion and may prove useful in an analysis of the mechanism of PAH transport system in renal basolateral membranes. PMID- 2992406 TI - The renin-angiotensin system and body function. AB - Angiotensin II, the biologically active component of the renin-angiotensin system, acts throughout the body to produce an impressive number of cardiovascular, endocrine, metabolic, and behavioral effects. Major actions include elevation of arterial pressure, stimulation of aldosterone secretion, and a variety of effects on the kidneys, brain, and pituitary. Investigation of the role of the renin-angiotensin system in physiological regulation has been greatly facilitated by the availability of specific inhibitors of the formation or actions of angiotensin II, most notably converting-enzyme inhibitors and angiotensin receptor antagonists. Studies with these agents have clearly shown that the renin-angiotensin system plays an important role in the defense of body balance and blood pressure in hypovolemic state, including sodium deficiency and hemorrhage. The inhibitors also lower blood pressure in some forms of hypertension, and converting-enzyme inhibitors are proving to be effective antihypertensive agents. PMID- 2992407 TI - Myotonic muscular dystrophy. From clinical description to molecular genetics. PMID- 2992408 TI - Effects of unilateral lesion of the inferior olive on L-[3H] aspartate receptors binding in synaptic membranes of cat cerebellar cortex. AB - In adult cats, local injection of kainic acid (KA) in the inferior olive (IO) of one side, from which the crossed olivocerebellar projection originates, produced asymmetric postural and motor deficits, attributed to selective damage of the olivary neurons. Since aspartate is one of the putative transmitters of the olivocerebellar fibers, experiments were performed to find out whether 6-8 days after injection of KA within the IO of one side produced changes in aspartate receptors binding in different zones of the cerebellar cortex. In particular, binding in the contralateral zones of the cerebellar cortex was referred to proteins contained in membrane suspensions and compared with the control values obtained in the same experiments from the ipsilateral zones. Binding of L-[3H] aspartate decreased on the average to 53.4% of the control value in the medial zone and to 86.1% of the control value in the intermediate and lateral zones of the cerebellar cortex. This reduction varied in different experiments according to the side of the injection, in agreement with the well known pattern of regional distribution of the olivocerebellar projection within the cerebellar cortex. These findings favour aspartate as the putative neurotransmitter of the climbing fibers. The demonstration that binding of aspartate decreased in the cerebellar cortex of one side, 6-8 days after injection of KA in the corresponding IO, indicates that plastic events occur at this level following destruction of the olivocerebellar pathway. In particular, the reduced binding can be attributed either to a decrease in number of the postsynaptic receptor sites for aspartate or to a decreased affinity of this amino acid for the corresponding receptors. These findings, however, do not exclude that an hypersensitivity by denervation may occur at the level of individual Purkinje cells when they are deprived of the climbing fibers input. In order to answer this question further experiments are required to find out how the binding for aspartate is modified at increasing time intervals after the olivary lesion. PMID- 2992409 TI - Temporal changes in mitral cell response patterns during repeated odor exposure. AB - The olfactory system of 17 goldfish was stimulated by natural odors, and from 31 mitral cells activity was extracellularly recorded as a response to the stimuli. 40 experiments were available for evaluation because nine of the 31 cells were investigated with respect to two different odors. The aim of this study was to examine the changes of the activity patterns during repeated runs of an experiment. 120 runs were taken in 15 experiments, 40 runs in 25 experiments, and 37 runs in 2 experiments. Nineteen out of the forty recordings showed patterned activities. In ten cases the patterns remained constant in all runs, while pattern changes occurred in nine cases. These changes sometimes happened abruptly after the first run or developed gradually over up to forty runs in other cases. Possible causes of the pattern changes are discussed. PMID- 2992410 TI - Electron microscopic investigation on the fine structure of the virus of enzootic bovine leukosis (BLV). AB - Systematically electron microscopical control of BLV producing cells and stimulated lymphocytes of leukotic cattle in the last years revealed maturation processes of BLV in a different form from comparable type C viruses. Electron microscopic representation of bridge-like junctions between core and envelope within examined retroviruses were discussed and compared with new model concepts. PMID- 2992411 TI - Calcium, calcium-ATPase and cyclic AMP changes in Ehrlich ascites cells submitted to hyperthermia. AB - Increased radiocalcium influx, decreased Ca-ATPase activity and cAMP concentration were observed in Ehrlich carcinoma cells submitted to hyperthermia temperature (45 degrees C). The significant augmentation of mitochondrial calcium that is simultaneous to an inhibition of the calcium pump appears to indicate a heat-induced failure of the system of calcium-extrusion that may be responsible for cell inactivation and death through cell toxic death by calcium overloading. The decreased cAMP concentration appears to corroborate the influx of calcium. PMID- 2992412 TI - [Cytological-histological case study of a dimorphous bronchial carcinoma]. AB - The case of a 55-year-old man who died of a dimorphic bronchial carcinoma is described. Diagnostic uncertainties and consequences resulting from the cytological diagnosis are discussed. PMID- 2992413 TI - Multiple islet cell adenomas in a patient with secondary hemochromatosis. PMID- 2992415 TI - Parathion-provoked lethality in rats is reduced by diethyldithiocarbamate. AB - Intoxication of male rats with 5 mg/kg parathion reduces survival to 15%. Diethyldithiocarbamate (DDC), known to inhibit mixed-function oxidase activity, has no effect on survival rate when this compound is used in a dosage of 300 mg/kg 45 min before or 10 min after parathion intoxication. However, when DDC and parathion are administered simultaneously, the rate of survival rises to 53%. PMID- 2992414 TI - Metabolism of protein supplements studied by the rumen simulation technique (Rusitec). AB - Long-term experiments were made with the Rumen Simulation Technique (Rusitec) with hay diet and with three different protein supplements (casein, fish meal and soyabean meal). Soyabean meal made the greatest contribution to the output of end products of fermentation and casein made the smallest contribution. The supplements did not have any marked effect on the metabolism of the basal diet of hay. The experiments showed that rumen microorganisms destroy a large proportion of protein (measured as total alpha-NH2 groups) and that this loss of protein was increased by supplementation with high protein compounds. The degradation of protein in soyabean meal and fish meal appeared to be considerably greater in the model system than the reported results of measurement in vivo and in situ. PMID- 2992416 TI - Neoplastic and nonneoplastic lesions in aging female rats with special reference to the functional morphology of the hyperplastic and neoplastic changes in the pituitary gland. AB - Hyperplastic and neoplastic lesions in the mammary glands, uterus, adrenals and pituitary glands of untreated aging female rats were evaluated. The most common lesions in the uterus, mammary glands and adrenal cortex were endometrial stromal polyps, fibroadenomas and nodular hyperplasias, respectively. A few malignant neoplasias were found in the uterus and mammary glands. The pituitary glands of the senile untreated rats had diffuse hyperplasia and/or hypertrophy of PRL, STH and ACTH cells. In 25 out of 52 rats (48%), pituitary adenomas were detected. Histological examination showed that the adenomas were made up of immature cells, a combination of ACTH and immature cells, prolactin cells, or intermediate zone cells. The cytological changes in the hypophyseal cell types in the senile rats were usually accompanied by spontaneous neoplasias of the mammary glands. The possible role of hypophyseal hormones in the neoplasias of the mammary glands and the adrenal cortex is discussed. The present study has also shown that the variation in the reproductive state of the aging female rats, as assessed from ovarian morphology, had no striking effect on the incidence or the severity of the hyperplastic/neoplastic lesions in the organs examined. PMID- 2992417 TI - Spread of herpes simplex virus and distribution of latent infection after intraocular infection of the mouse. AB - Intraocular inoculation of HSV 1 in the mouse results not only in uveitis, but also in the spread of virus via sensory, sympathetic and optic nerves. During the acute infection with HSV 1 strain SC 16 in both outbred and NIH (inbred) mice, virus reached the ipsilateral trigeminal ganglion, superior cervical ganglion, both sides of the brain stem and the contralateral (uninoculated) eye. With HSV 1 strain KOS in outbred mice the same tissues became infected but virus was also isolated from the ophthalmic part of the contralateral trigeminal ganglion. After resolution of the acute disease in outbred mice, latent infection with strain KOS was demonstrated in both trigeminal ganglia and in the ipsilateral superior cervical ganglion. With strain KOS, virus was sometimes isolated from eyes removed more than a month after inoculation and then cultured in vitro for 2-3 weeks. By electron microscopy infected cells were seen in the choroid and sclera of such eyes. PMID- 2992418 TI - Morphology and immunoelectron microscopy of AIDS virus. AB - Lymphocytes infected with human T-lymphotropic virus type III/lymphadenopathy associated virus were examined by thin-section and immunoelectron microscopy, with horseradish peroxidase as an electron-dense marker. There was no evidence of an intracytoplasmic virion precursor, but viral antigen accumulated at the plasma membrane. Particles formed at the plasma membrane, and most virus was found in extracellular spaces. Mature virus had an envelope with fuzzy surface projections surrounding a centric core and an electron-dense eccentric nucleoid. PMID- 2992419 TI - Distinction of viral and host-derived glycopolypeptides induced by "early" functions of human cytomegalovirus. AB - Human cytomegalovirus is shown to induce in phosphonoacetic acid-treated human fibroblasts glycosylation of five polypeptides with approximate molecular weights of 200-250, 150, 135, 130 and 100 kilodaltons (kd). Except for the 130 kd product, these glycopolypeptides (gp) separate with the cytoplasmic fraction, only one (200-250 kd) with the chromatin fraction as well. The gp of 135 and 100 kd were found to be virus-specified as determined by immunoblotting and immunoprecipitation. The gp of 200-250 kd exhibited an immunological relatedness to fibronectin and are therefore considered host-specific products. Both subsets of gp participate in virus-induced surface membrane alterations as documented by living cell immunofluorescence. PMID- 2992420 TI - Isolation and studies of myxovirus glycoproteins. AB - The isolation of ortho- and paramyxovirus glycoproteins using a new nonionic detergent (MESK) is reported. MESK was shown to solubilize most of the viral envelope glycoproteins without decreasing their biologic activity. Solubilized glycoproteins are not contaminated by any internal viral proteins or by appreciable quantities of viral envelope lipids. The removal of MESK by dialysis resulted in the formation of glycoprotein micelles. The immunogenic activity of isolated glycoproteins was compared to that of virus particles. Immunization with isolated glycoproteins was shown to protect mice against a lethal influenza infection. Virions were treated with MESK in the presence of exogenous egg phosphatidylcholine, detergent was removed by dialysis and the glycoprotein was reconstituted in the vesicles. This reconstitution was accompanied by restoration of the haemolytic activity of Sendai virus proteins up to that of native virus particles. The level of activity, also the morphology and buoyant density of the vesicle were dependent on the protein/lipid ratio. MESK proved to be of value for the selective solubilization of the surface glycoproteins of animal enveloped viruses and their reconstitution in liposomes. PMID- 2992421 TI - Macropodid herpesviruses 1 and 2: two herpesviruses from Australian marsupials differentiated by restriction endonucleases, DNA composition and hybridization. Brief report. AB - The DNAs of a number of herpesviruses from Australian marsupials have been analyzed using restriction endonucleases, dot-blot hybridizations, and analytical ultracentrifugation in CsCl. The data clearly show the presence of 2 distinct viruses, designated macropodid herpesviruses 1 and 2. PMID- 2992422 TI - Entry of adult T-cell leukemia-associated virus into human peripheral blood leukocytes. Brief report. AB - Within 10 minutes after tritium-labeled adult T-cell leukemia-associated virus (ATLV) inoculation, silver grains were found over human lymphocytes. At the time of entry of ATLV, the viral envelope was observed to fuse with the cell membrane. PMID- 2992424 TI - [Additional findings on contact sensitivity of mice induced by toluene diisocyanate (TDI)]. PMID- 2992423 TI - [Plasma prostaglandin and thromboxane levels in asthmatic children]. PMID- 2992426 TI - Radiculopathy and electromyography. PMID- 2992425 TI - [Beta-adrenergic receptor in bronchial asthma. Part 2. Human placental beta adrenergic receptor in normal and asthmatic subjects]. PMID- 2992427 TI - Legionnaires' disease-associated neuropathy. PMID- 2992428 TI - Status epilepticus and Epstein-Barr virus encephalopathy. Diagnosis by modern serologic techniques. AB - Three patients had status epilepticus appearing de novo as the presenting manifestation of Epstein-Barr virus (EBV) encephalopathy. Clinical findings of infectious mononucleosis were absent and EBV-specific serologic tests made or confirmed the diagnosis in each case. Epstein-Barr virus should be considered as a potential cause when status epilepticus occurs in the absence of previous seizures or an identified cause of seizures. The diagnosis may be made in some cases only with the use of EBV-specific serologic testing. PMID- 2992429 TI - Adenovirus type 2 encephalitis and concurrent Epstein-Barr virus infection in an adult man. AB - A 30-year-old bisexual male prisoner with a history of intravenous drug abuse manifested encephalitis. Adenovirus type 2 was isolated from brain tissue obtained by a biopsy and there was a concurrent fourfold rise in the antibody titer. In addition, an initial negative result from a slide test for infectious mononucleosis heterophil antibodies (Monospot) test converted to positive and there was an increase in the IgG antibody titer to Epstein-Barr virus (EBV) viral capsid antigen. Although he was anergic to skin test antigens, T-cell subsets and lymphocyte transformation study results were normal. This case demonstrated a rare adenovirus encephalitis with a simultaneous EBV infection in a patient at risk for acquired immunodeficiency syndrome, who had minimal evidence of T-cell deficiency. PMID- 2992430 TI - Adsorption of human salivary mucins to hydroxyapatite. AB - The interactions between low (MG2) and high (MG1) molecular-weight human submandibular-sublingual mucin and hydroxyapatite (OHAp) were compared using a quantitative assay. Data obtained appeared to empirically fit the Langmuir adsorption isotherm. Apparent affinity constants derived from this isotherm indicated that MG1 had a greater affinity for OHAp than did MG2. Inhibition studies revealed that salivary glycolipids inhibited the interaction of MG1 and OHAp without influencing MG2 adsorption. In contrast, MG2 adsorption to OHAp was markedly inhibited by cysteine-containing salivary phosphoprotein fractions. Collectively, the data indicate MG1 and MG2 differ in their interaction with OHAp. PMID- 2992431 TI - Pathologic diagnosis: pleomorphic adenoma. PMID- 2992432 TI - An investigation of vertical transmission of a fowl adenovirus serotype 8. PMID- 2992433 TI - A rapid procedure for the purification of avian encephalomyelitis viruses. AB - A rapid procedure for the purification of egg-grown or field preparations of avian encephalomyelitis virus (AEV) of neural origin is described. Extracts of infected tissues were clarified and then partly purified with trichlorotrifluorethane (Freon TF), and the virus present was concentrated with polyethylene glycol. The concentrates were then re-extracted with Freon, and a portion was labeled with 125iodine. During subsequent purification steps, virus could be readily detected by monitoring for radioactivity, thus eliminating the need to determine the infectivity in individual fractions or to examine for the presence of virions by electron microscopy. Final purification was achieved by cesium-chloride equilibrium or sucrose-velocity-gradient centrifugation. Virus purified in this manner was shown to be free of tissue debris, to be specific for AEV by immune electron microscopy, and to possess structural proteins characteristic of picornaviruses. PMID- 2992435 TI - Identification of field isolates of infectious bronchitis virus by interference with the La Sota of strain of Newcastle disease virus. AB - The interference phenomenon of infectious bronchitis virus (IBV) with growth of Newcastle disease virus (NDV) in embryonating chicken eggs (ECE) was used as a diagnostic method. Fifteen field isolates obtained from presumptively infectious bronchitis-affected chickens were analyzed by the IBV-NDV interference test. Eight isolates were capable of interfering with the growth of the La Sota strain of NDV, as measured by hemagglutination (HA) activity when IBV was inoculated 10 hr before NDV into ECE. The interference was considered specific for IBV, because it could be eliminated by adding homologous anti-IBV serum. The sensibility of this method could be demonstrated, because in some cases low-passage levels of IBV isolates showing HA interference ability were not capable of producing lesions in ECE. Furthermore, serologically negative IBV samples did not interfere with NDV growth. From these results, the IBV-NDV interference test appears to be a potential diagnostic alternative for identifying IBV field isolates. PMID- 2992436 TI - Hemorrhagic enteritis of turkeys in California: serologic study of hemorrhagic enteritis virus antibody with an enzyme-linked immunosorbent assay. AB - The incidence of hemorrhagic enteritis (HE) infection in California turkeys was studied by testing 2220 turkey blood samples from 173 flocks for HE virus (HEV) antibody by the enzyme-linked immunosorbent assay (ELISA). Maternal antibody was detected at 1 day of age in all flocks tested, and it vanished after 3 weeks. Acquired HEV antibody appeared at 8 to 10 weeks, and 100% of the meat and breeder turkey flocks were positive after 11 weeks of age. HEV infection occurred earlier in the meat flocks than in the breeder flocks, and it also occurred earlier during summer than during the fall and winter months. PMID- 2992434 TI - Efficacy of experimental inactivated mycoplasma gallisepticum oil-emulsion bacterin in egg-layer chickens. AB - Six groups of white leghorn pullets were studied to determine the ability of beta propiolactone-inactivated Mycoplasma gallisepticum (MG) oil-emulsion bacterins to counteract reductions in egg production caused by MG infection. The pullets were inoculated with 0.5 ml of MG bacterin subcutaneously in the neck at about 20 weeks of age and were challenged with MG near 28 weeks of age, when they were in peak egg production. Various challenge schemes with infectious bronchitis virus were used at the time of MG challenge to increase the reduction in egg production. MG bacterins afforded protection against moderate drops in egg production in at least three of the studies, where the unvaccinated challenged control hens exhibited reduced egg production. PMID- 2992437 TI - Exacerbative effect of vitamin A on malabsorption syndrome in chicks. AB - The interaction between malabsorption syndrome (MAS) and dietary vitamins A and D was studied in broiler chicks reared in floor pens for 4 weeks. The chicks were naturally infected with MAS, whereas hatchmates fed the same diets but in a separate facility (battery brooder) did not exhibit signs of MAS and, therefore, were considered controls. MAS significantly reduced body weights, bone ash, serum calcium and phosphorus concentrations, and liver lipids and increased the incidence of skeletal abnormalities (tibial dyschondroplasia and rickets). Rather than ameliorating the effects of MAS, vitamin A caused a further reduction in body weight and bone ash. A possible nutrient interaction between vitamin A and vitamin D or vitamin E in birds with MAS may account for the exacerbative effect of vitamin A. PMID- 2992438 TI - Use of egg yolk to determine antibody levels in chickens inoculated with a hemagglutinating duck adenovirus (adenovirus 127-like). AB - Chickens were experimentally infected with a duck adenovirus that has been shown to be serologically indistinguishable from Adenovirus 127. Sera and eggs were collected at intervals after exposure for antibody determination by the hemagglutination-inhibition (HI) test, the enzyme-linked immunosorbent assay (ELISA), and the immunodiffusion (ID) test. Egg yolks were processed for use in the serological tests by (a) dilution in phosphate-buffered saline (PBS), (b) extraction of the water-soluble fraction with chloroform, or (c) freezing and thawing PBS-diluted yolks and testing the supernatant fluid. HI antibody titers from serum and extracted yolk were similar except during the initial 2 weeks, when yolk antibody levels were low or absent. Chloroform-extracted yolks were suitable material for the HI, ELISA, and ID tests. Heat inactivation of the chloroform-extracted yolk had no effect on titers. PMID- 2992439 TI - Concurrent infections with reoviruses and coccidia in broilers. AB - These experiments investigated the interaction among two species of coccidia (Eimeria acervulina and E. mitis) and three strains of reovirus (virus 2035, a weak to moderate pathogen; and viruses 2408 and 1733, severe pathogens). When reoviruses were not present, high inoculation dosages (10(6) sporulated oocysts/bird) of both E. acervulina and E. mitis depressed weight gain, plasma pigment, and plasma protein. Low doses of coccidia (10(4) oocysts) in the absence of virus had no such effect on weight gain. When high doses of coccidia were present at the same time as virus 2035 or 2408, they resulted in a significantly greater depression of weight gain than when either virus or coccidia were present alone. With virus 2035, this greater depression was seen even when low doses of coccidia were used. Lesion scores due to coccidiosis and the number of oocysts produced were not affected by previous exposure to reovirus. Both coccidiosis and reovirus infections increased the frequency of some leg problems and other abnormal conditions. The most obvious interaction between coccidia and reovirus was the marked increase in swollen hocks seen when coccidia and virus 2035 were present together (20-27%) compared with either the virus or coccidia alone (0 10%). Virus 2408 interfered slightly with the development of immunity to coccidia. There was some indication that early coccidiosis could increase the ability of some virus isolates to infect various tissues of the host. PMID- 2992440 TI - Isolation and characterization of avian rotaviruses. AB - Rotaviruses were isolated from intestinal contents obtained from flocks of turkey poults and pheasant poults with diarrhea and from different age groups of chickens showing various signs of intestinal disorders. The incorporation of 5 micrograms trypsin/ml in the inoculum and medium was essential for virus isolation in chicken kidney cells. All isolates were identified as rotaviruses by fluorescent-antibody technique using a National Institutes of Health reference rotavirus antiserum against human rotavirus strain "D," Type 2. Negative-staining and transmission electron microscopy showed the presence of rotavirus particles. Furthermore, polyacrylamide gel electrophoresis pattern of viral RNA segments of our isolates confirmed that they are rotaviruses. Seven-day-old turkey poults could be infected with turkey and chicken rotavirus isolates; in contrast, chicks of the same age were refractory. PMID- 2992441 TI - Isolation of infectious bronchitis virus from intestine and reproductive organs of laying hens with dropped egg production. AB - A total of 52 samples (22 cloacal swabs and 30 pooled ovaries and oviducts) from hens with a history of a 25 to 40% drop in egg production from 30 flocks were examined for the isolation of infectious bronchitis virus in embryonated chicken eggs. Five isolates were obtained: three from cloacal swabs and two from reproductive organs. These isolates were identified and characterized on the basis of agar-gel-precipitation, hemagglutination, and neutralization tests and other physicochemical characteristics. PMID- 2992442 TI - Comparative use of direct organ cultures of infected chicken tracheas in isolating avian infectious bronchitis virus. AB - Five trials were conducted to compare four in vitro methods of isolating avian infectious bronchitis virus (IBV)-direct organ culture of infected tracheal rings (DOC), inoculation of tracheal organ culture (OC), inoculation of chicken embryo, and inoculation of cultured cells. DOC was prepared from tracheas of chickens experimentally inoculated with field samples. In the other methods, pooled tracheal and kidney suspensions were used to inoculate OC, chicken embryos, and cultured cells. IBV was consistently isolated at the initial passage by the DOC and OC inoculation systems, but it was not always isolated by embryo inoculation and never isolated by cultured-cell inoculation. When combined with immunofluorescent staining, DOC was much more efficient than the OC inoculation system for isolation and identification of the five strains of IBV tested because of its simplicity and speed. PMID- 2992443 TI - Effect of avian reoviruses on lymphoid organ weights and antibody response in chickens. AB - Several avian reoviruses were screened to determine their effects on the immune system by inoculating them subcutaneously (SQ) into day-old chicks. For comparison, infectious bursal disease virus (IBDV) was similarly evaluated. The response of the immune system was measured functionally by the hemagglutination inhibition (HI) response to Newcastle disease virus (NDV) and structurally by changes in the organ-to-body-weight ratios of the bursa of Fabricius, thymus, and spleen. When inoculated SQ, most of the reoviruses caused transient alterations in lymphoid organ weights, decreasing the bursa weight and increasing the spleen weight. Of those reoviruses tested, only one--a commercial vaccine based on the isolate S-1133--demonstrated the ability to interfere significantly with NDV-HI responses, although several had numerically lower titers. Two of the isolates were also evaluated by oral inoculation. Giving the viruses orally did not cause any alterations in organ weights; however, both isolates depressed the HI response of chicks to NDV. Compared with reoviruses, IBDV significantly depressed NDV-HI titers. The structural responses to IBDV differed, however: IBDV significantly depressed bursa weights for all 3 weeks of the test period without affecting spleen weights. Some of the reovirus isolates inoculated SQ were lethal for day-old chicks. This, and their ability to alter the lymphoid-to-body-weight ratios of the spleen and bursa, could be considered valid criteria by which to study the pathogenesis of these agents. PMID- 2992444 TI - Necrotizing hepatitis associated with a reo-like virus infection in a parrot. AB - A young African grey parrot was presented for necropsy following a 3-to-4-day illness consisting of lethargy, anorexia, and diarrhea. A disseminated necrotizing hepatopathy was evident upon histologic examination of the liver. Aggregates of approximately 70-nm-diameter viral particles having morphology consistent with the family Reoviridae were demonstrated by thin-section and negative-staining electron microscopy. PMID- 2992445 TI - Dihydroergotoxine, an ergot alkaloid without marked hypotensive effect, improves blood flow and metabolism in the underperfused canine myocardium. AB - Regional myocardial blood flow was assessed in 6 anaesthetized dogs using radiolabelled microspheres. Lactate, H+, and O2 content were measured in arterial and local venous blood, obtained from the area supplied by the circumflex branch of the left coronary artery (LCX). Constriction of LCX by 67% led to considerable hemodynamic changes indicative of depressed global myocardial function. A significant release of lactate from the underperfused part of the left ventricle was recorded. Infusion of 0.7 micrograms/kg dihydroergotoxine over a period of 5 min induced a decrease in heart rate, left ventricular dp/dt max, and cardiac output, a sustained increase in total peripheral and femoral resistance and a transient increase in mean arterial blood pressure. Dihydroergotoxine markedly reduced lactate release and even, in some cases, induced lactate uptake and led to a significant increase in the endocardial blood flow both in the underperfused and normally perfused left ventricle. It is concluded that dihydroergotoxine, due to its ability to reduce myocardial oxygen demand and owing to its alpha sympatholytic activity, which is not accompanied by any marked decrease in arterial blood pressure, causes improvement in blood flow in the underperfused myocardium and a marked amelioration of the impaired metabolic processes. PMID- 2992447 TI - [Replication characteristics and cytopathic effect of southern German field isolates of chicken infectious bronchitis virus in cell cultures and the effect of trypsin]. PMID- 2992448 TI - [A mixed mamma tumor, a theca cell tumor and a leiomyoma in a bitch]. PMID- 2992446 TI - Ca overload and the action of calcium sensitive proteases, phospholipases and prostaglandin E2 in myocardial cell degradation. AB - The mechanism of Ca overload-induced myocardial cell injury under hypoxia was examined for the involvement of calcium-activated neutral proteases (CANP), calcium-dependent phospholipases (CDP) or prostaglandins with measuring 45Ca entry, intake of biologically inert dye, nigrosin, into the cultured myocytes, as was useful for the quantification of sarcolemma permeability, and the release of creatine phosphokinase (CPK) to the culture medium. A Ca channel blocker, verapamil (1 and 10 micrograms/ml) or a Ca ionophore, A23187 (0.5 to 4 micrograms/ml) dose-dependently decreased or increased both the Ca entry and nigrosin intake in accordance with the CPK release. Furthermore, the inhibitors against CANP, NCO-700 (2 and 20 micrograms/ml) that was demonstrated to permeate sarcolemma using 14C-labelled reagent, against CDP, mepacrine (1 and 10 micrograms/ml) or against cyclooxygenase, indomethacin (1 and 10 micrograms/ml) caused no effect on the Ca entry, nigrosin intake nor CPK release under hypoxia. These results suggest that the Ca overdose into the myocardial cells potentiates their injury and it is not primarily related to the activation of CANP, CDP nor cyclooxygenase. PMID- 2992449 TI - Some properties of cellobiose oxidase from the white-rot fungus Sporotrichum pulverulentum. AB - Cellobiose oxidase from the white-rot fungus Sporotrichum pulverulentum has been purified to homogeneity by a new procedure. The carbohydrate and amino acid compositions of the enzyme have been determined. Cellobiose oxidase contains FAD and cytochrome b prosthetic groups. Mr of the enzyme has been estimated at 74400 by sedimentation equilibrium. The enzyme is a monomer. Optical, fluorescence and e.p.r. spectra of oxidized and reduced cellobiose oxidase have been determined. A preliminary investigation of the substrate specificity of cellobiose oxidase reveals that disaccharides and even some insoluble polysaccharides are substrates, but not monosaccharides. Strong substrate inhibition is seen at high concentrations of cellobiose. This effect is particularly marked when oxygen is the electron acceptor. Cellobiose oxidase is unusual among flavoproteins, since it stabilizes the red anionic flavin semiquinone and forms a sulphite adduct, yet appears to produce the superoxide anion as its primary reduced oxygen product. PMID- 2992450 TI - Production of the superoxide adduct of myeloperoxidase (compound III) by stimulated human neutrophils and its reactivity with hydrogen peroxide and chloride. AB - Examination of the spectra of phagocytosing neutrophils and of myeloperoxidase present in the medium of neutrophils stimulated with phorbol myristate acetate has shown that superoxide generated by the cells converts both intravacuolar and exogenous myeloperoxidase into the superoxo-ferric or oxyferrous form (compound III or MPO2). A similar product was observed with myeloperoxidase in the presence of hypoxanthine, xanthine oxidase and Cl-. Both transformations were inhibited by superoxide dismutase. Thus it appears that myeloperoxidase in the neutrophil must function predominantly as this superoxide derivative. MPO2 autoxidized slowly (t 1/2 = 12 min at 25 degrees C) to the ferric enzyme. It did not react directly with H2O2 or Cl-, but did react with compound II (MP2+ X H2O2). MPO2 catalysed hypochlorite formation from H2O2 and Cl- at approximately the same rate as the ferric enzyme, and both reactions showed the same H2O2-dependence. This suggests that MPO2 can enter the main peroxidation pathway, possibly via its reaction with compound II. Both ferric myeloperoxidase and MPO2 showed catalase activity, in the presence or absence of Cl-, which predominated over chlorination at H2O2 concentrations above 200 microM. Thus, although the reaction of neutrophil myeloperoxidase with superoxide does not appear to impair its chlorinating ability, the H2O2 concentration in its environment will determine whether the enzyme acts primarily as a catalase or peroxidase. PMID- 2992452 TI - The mechanism of guanosine triphosphate depletion in the liver after a fructose load. The role of fructokinase. AB - A Sephadex G-25 filtrate of a 100 000g supernatant of rat liver homogenate was shown to be able to phosphorylate fructose, with GTP as the phosphate donor. Attempts to separate ATP- and GTP-dependent fructokinase activities failed, indicating that there is a single enzyme able to use both nucleotides. With a partially purified enzyme, Km values for fructose of 0.83 and 0.56 mM were found with ATP and GTP as substrates respectively. Km values of 1.53 and 1.43 mM were found for GTP and ATP respectively. Both ADP and GDP inhibited the GTP- and ATP dependent fructokinase activity. We conclude that the depletion of hepatic GTP caused by intravenous administration of fructose to mice and rats can be explained simply by the utilization of the nucleotide by fructokinase. PMID- 2992451 TI - Simultaneous purification and characterization of glucokinase, fructokinase and glucose-6-phosphate dehydrogenase from Zymomonas mobilis. AB - The three enzymes glucokinase (EC 2.7.1.2), fructokinase (EC 2.7.1.4) and glucose 6-phosphate dehydrogenase (EC 1.1.1.49) were isolated in high yield from extracts of Zymomonas mobilis. The principal steps in the isolation procedures involved the use of selected dye-ligand adsorbent columns, with affinity elution of two of the three enzymes. Glucokinase and fructokinase are dimeric proteins (2 X 33000 Da and 2 X 28000 Da respectively) and glucose-6-phosphate dehydrogenase is a tetramer (4 X 52000 Da). Some similarities in the structural and kinetic parameters of the two kinases were noted, but they have absolute specificity for their substrates. Fructokinase is strongly inhibited by glucose; otherwise non substrate sugars had little effect on any of the three enzymes. PMID- 2992454 TI - Digitonin-collagenase perfusion for efficient separation of periportal or perivenous hepatocytes. AB - Intact rat liver cells from the perivenous region were isolated by collagenase perfusion after first destroying the periportal region by a brief portal infusion of digitonin. Periportal cells were isolated after retrograde digitonin infusion. Significantly higher alanine aminotransferase, gamma-glutamyltransferase and lactate dehydrogenase activities and lower glutamate dehydrogenase and pyruvate kinase activities in periportal than in perivenous cells demonstrate marked separation. The high yield allows further characterization in vitro of the cell populations. PMID- 2992453 TI - Studies on the role of inositol trisphosphate in the regulation of insulin secretion from isolated rat islets of Langerhans. AB - Glucose (20 mM) and carbachol (1 mM) produced a rapid increase in [3H]inositol trisphosphate (InsP3) formation in isolated rat islets of Langerhans prelabelled with myo-[3H]inositol. The magnitude of the increase in InsP3 formation was similar when either agent was used alone and was additive when they were used together. In islets prelabelled with 45Ca2+ and treated with carbachol (1 mM), the rise in InsP3 correlated with a rapid, transient, release of 45Ca2+ from the cells, consistent with mobilization of 45Ca2+ from an intracellular pool. Under these conditions, however, insulin secretion was not increased. In contrast, islets prelabelled with 45Ca2+ and exposed to 20mM-glucose exhibited a delayed and decreased 45Ca2+ efflux, but released 7-8-fold more insulin than did those exposed to carbachol. Depletion of extracellular Ca2+ failed to modify the increase in InsP3 elicited by either glucose or carbachol, whereas it selectively inhibited the efflux of 45Ca2+ induced by glucose in preloaded islets. Under these conditions, however, glucose was still able to induce a small stimulation of the first phase of insulin secretion. These results demonstrate that polyphosphoinositide metabolism, Ca2+ mobilization and insulin release can all be dissociated in islet cells, and suggest that glucose and carbachol regulate these parameters by different mechanisms. PMID- 2992455 TI - Influence of divalent cations on the detection of somatogenic and lactogenic binding sites in mouse liver cells. AB - Specific binding of 125I-labelled human somatotropin was demonstrated in isolated hepatocytes from male mice. In the presence of divalent cations (Ca2+ and Mg2+) the binding of 125I-labelled human somatotropin was competitive with ovine prolactin. Scatchard analysis of competition data indicated a KD of 1.4 +/- 0.2 nM and a binding capacity of 13 000 +/- 2000 sites/cell. In the absence of divalent cations and in the presence of EDTA, human and bovine somatotropins were found to be equally effective to displace bound 125I-labelled human somatotropin, while ovine prolactin showed a weak competition. In this case, the binding capacity was 8400 +/- 1500 sites/cell and the KD was 1.1 +/- 0.1 nM. PMID- 2992456 TI - Glycolytic and gluconeogenic states in an enzyme system reconstituted from phosphofructokinase and fructose 1,6-bisphosphatase. AB - Transitions between glycolytic and gluconeogenic states have been investigated in an open and homogeneous enzyme system containing phosphofructokinase, fructose 1,6-bisphosphatase, pyruvate kinase, adenylate kinase and glucose 6-phosphate isomerase. The direction of substrate flow was found to depend on the maximum activities of phosphofructokinase and fructose 1,6-bisphosphatase as well as on the influx concentrations of fructose 6-phosphate and fructose 1,6-bisphosphate. At high and low maximum activities of phosphofructokinase and fructose 1,6 bisphosphatase unique and stable stationary states occur, whereas at intermediate enzyme concentrations sustained oscillations emerge. Stationary states with a low rate of substrate cycling demand both appropriate enzyme concentrations and an adequate substrate supply. Accordingly, transitions between efficient glycolytic and gluconeogenic states require changes of the enzyme concentrations and of the supply of substrates. Such transitions exhibit a transient oscillatory response. The sustained oscillations generated at intermediate activities of phosphofructokinase and fructose 1,6-bisphosphatase lead to a significant diminution of the rate of substrate cycling when compared with the respective steady state values. During the oscillations glycolytic and gluconeogenic states are consecutively passed through. Because of this a temporal organization of the antagonistic reactions is achieved. In our system the kinetic organization of the two opposite reactions is mainly brought about by the reciprocal allosteric effects of AMP on the activities of the two enzymes. PMID- 2992457 TI - Purification of DNA from formaldehyde fixed and paraffin embedded human tissue. AB - The ability to isolate DNA from preserved human tissues would provide numerous experimental opportunities. In this report it is shown that DNA can be extracted from tissues prepared for routine histopathological examination (i.e., fixed with formaldehyde and embedded in paraffin). Although the extracted DNA is not intact, it is double stranded, cleavable with restriction endonucleases, and suitable for a variety of standard techniques used in molecular biology. PMID- 2992458 TI - Effect of reduced oxygen intermediates on sarcolemmal muscarinic receptors from canine heart. AB - The effect of oxygen free radicals generated by xanthine-xanthine oxidase system and hydrogen peroxide were investigated on cardiac muscarinic cholinergic receptors. We have used highly enriched sarcolemmal preparations isolated from canine myocardium. Exposure of the sarcolemma to oxygen free radicals by xanthine xanthine oxidase system resulted in a significant (P less than 0.05) decrease of Bmax of (3H)-QNB (4.66 +/- 0.51 to 2.68 +/- 0.22 pmoles/mg protein). Addition of superoxide dismutase (SOD) and catalase (10 micrograms/ml) resulted in a significant reversal of Bmax value to 3.72 +/- 0.39 pmoles per mg protein (p less than 0.05). However, the affinity constants of dissociation (KD) were not altered appreciably with the exposure to oxygen free radicals with or without scavengers. Hydrogen peroxide significantly depressed 3H-QNB binding to the receptors in a dose-dependent manner in a concentration range between 4.41 mM -441 mM. This depression was completely inhibited by 10 micrograms/ml catalase. The study demonstrates that the oxygen free radical species are capable of disrupting (3H) QNB binding to the cardiac muscarinic receptors. PMID- 2992459 TI - A 25 000 dalton inhibitor of cAMP independent protein kinases present in rat liver HMG protein preparations. AB - A protein kinase inhibitor was found in rat liver cells as a component of HMG proteins. It is located in cytosol as well as in nuclei. It inhibits all tested cAMP independent protein kinases and has no effect on cAMP dependent protein kinases. This inhibitor is a 25 000 Da protein. It has no ATPase, phosphoprotein phosphatase or proteinase activity and is heat unstable. PMID- 2992460 TI - The role of reactive oxygen compounds derived from 6-hydroxydopamine for bone marrow purging from neuroblastoma cells. AB - 6-Hydroxydopamine(6-OHDA), a specific neurotoxin against sympathetic nerve cells, is a drug already used for purging of bone marrow from neuroblastoma cells before autologous bone marrow transplantation. However, we could not detect significant differences in the toxicity of 6-OHDA against neuroblastoma and other tumor cells under the purging conditions clinically used. In contrast, bone marrow stem cells were much more resistant. The unspecific toxic effect of 6-OHDA is caused by H2O2 or H2O2-derived products which are generated by auto-oxidation in the incubation medium before a significant amount of 6-OHDA is taken up by the cells. Withdrawal of oxygen during the incubation period and subsequent incubation with an oxygen containing medium led to a more specific destruction of neuroblastoma cells which can take up 6-OHDA selectively. PMID- 2992461 TI - In vivo metabolism of leukotriene C4 in man: urinary excretion of leukotriene E4. AB - Five - 20 nmoles of [5,6,8,9,11,12,14,15-3H8]leukotriene C4 was injected into three male volunteers. Forty-eight percent of the administered 3H was recovered from urine and 8% from feces, within a 72 hr period. Of the total urinary radioactivity 44% was excreted during the first hour after injection. This activity was mainly found in one compound, designated "I". The radioactivity excreted into urine later than one hour after injection, consisted partly of Compound I and two additional components, and partly of polar, non-volatile material. Compound I was identified as leukotriene E4 by UV-spectroscopy and cochromatographies in three high performance liquid chromatography systems with synthetic reference compounds. A total of 13% of administered radioactivity was excreted in urine as leukotriene E4. PMID- 2992462 TI - Effects of calmodulin and related proteins on the hemolytic activity of melittin. AB - The calcium-dependent binding of melittin by calmodulin effectively inhibits the hemolytic activity of melittin in suspensions of washed rabbit erythrocytes. Protection is also obtained with troponin C (+/-Ca++), denatured phosphorylase kinase, and denatured calcineurin but not with whole troponin or the native enzymes. These effects can be used both in assays for melittin in venom samples and in determinations of calmodulin or related proteins. PMID- 2992464 TI - Topoisomerase-II activity in human leukemic and lymphoblastoid cells. AB - Topoisomerase-II activity was analyzed in various human leukemic and lymphoblastoid cell-lines with comparison to normal human peripheral blood lymphocytes. All of the examined tumor cells contained this enzyme in both the nuclear and cytoplasmic fractions, whereas no appreciable activity of the enzyme was detected in either fraction of the resting normal lymphocytes. Using pBR322 plasmid as a substrate, undialyzed extracts of the tumor cells exhibited the typical ATP-dependent relaxation of supercoiled circles and formation of linear and catenated structures, as well as the ATP-independent knotting activity. On the other hand, dialyzed extracts exerted only the ATP-dependent supercoil relaxation. Novobiocin inhibited the linearization and catenation but not the supercoil relaxing or knotting activities. This study provides indications for an excessive level of a structurally abnormal topoisomerase-II in these tumor cell lines. PMID- 2992463 TI - The amino acid sequence of the fluorescein-labeled peptides of electric ray and brine shrimp (Na,K)-ATPase. AB - (Na,K)-ATPase from Torpedo californica (electric ray) and Artemia salina (brine shrimp) was labeled with fluorescein 5'-isothiocyanate (FITC) with concomitant loss of activity. Both inactivation and binding were inhibited in the presence of ATP. The sequence of the peptide resulting from tryptic digest containing labeled lysine from both enzymes is Tyr-Leu-Leu-Val-Met-Lys*-Gly-Ala-Pro-Glu-Arg. Thus the primary structure of this region is shown to be conserved in the enzymes of a nonvertebrate and a vertebrate. PMID- 2992465 TI - Comparison of dipeptidyl carboxypeptidase and endopeptidase activities in the three enkephalin-hydrolysing metallopeptidases: "angiotensin-converting enzyme", thermolysin and "enkephalinase". AB - Angiotensin-converting enzyme (ACE), thermolysin and "enkephalinase", three metallopeptidases cleaving the Gly3-Phe4 amide bond of enkephalins, were compared regarding substrate specificity and effects of butanedione, an arginyl-directed reagent. The hydrolysis of enkephalins and analogues was more affected by the nature of P1 and P2 residues in the case of thermolysin than in those of ACE or "enkephalinase"; amidation of the C-terminal carboxylate decreased drastically the hydrolysis by ACE but only marginally by thermolysin and the effect was intermediate for "enkephalinase". With adequate model substrates, the ratio of dipeptidylcarboxypeptidase to tripeptidylcaroxypeptidase (endopeptidase) activities were of 25 for ACE, 3 for "enkephalinase" and only 0.3 for thermolysin. Finally a butanedione treatment increased thermolysin activity, but abolished ACE activity; it reduced "enkephalinase" activity by 80% when measured with a free C-terminal carboxylate enkephalin analogue but only slightly with the corresponding amidated derivative. A critical role of an Arg residue in ACE and, to a lesser extent, in "enkephalinase" (but not in thermolysin) is suggested to be responsible for the preferential dipeptidylcarboxypeptidase activity of these two enzymes. PMID- 2992466 TI - Synergistic induction of ornithine decarboxylase by diacylglycerol, A23187, and cholera toxin in guinea pig lymphocytes. AB - When guinea pig lymphocytes were cultured with 1-oleoyl-2-acetyl-glycerol (OAG), A23187, and cholera toxin, ornithine decarboxylase activity was induced synergistically, peaking at 6 h. Addition of 12-O-tetradecanoyl-phorbol 13 acetate (TPA), A23187, and dibutyryl cAMP caused the same kind of induction. Cholera toxin potentiated the ability of A23187 to induce ornithine decarboxylase, but not that of OAG. Dibutyryl cAMP augmented the induction caused by A23187 but not by TPA. These results suggest that both the activation of Ca++ sensitive, phospholipid-dependent protein kinase (protein kinase C) and the increase in intracellular levels of Ca++ and cAMP are necessary for this induction. cAMP may potentiate the induction by modulating a Ca++ messenger system other than that for protein kinase C activation. PMID- 2992467 TI - Studies on rhodanese synthesis in bovine adrenocortical cells. AB - The synthesis of adrenodoxin, a mitochondrial iron-sulfur protein required for adrenocortical steroidogenesis, is known to be regulated chronically by ACTH. Rhodanese, also a mitochondrial enzyme, is thought to be required for synthesis of iron-sulfur centers, such as those contained in adrenodoxin. In this study it has been found that rhodanese synthesis and activity are not regulated by ACTH, under the same conditions whereby ACTH induces adrenodoxin synthesis. In addition, unlike adrenodoxin, rhodanese is found to be synthesized in the mature form rather than as a higher molecular weight precursor protein. PMID- 2992468 TI - P450cam gene cloning and expression in Pseudomonas putida and Escherichia coli. AB - The gene camC, which encodes the cytochrome P450 monoxygenase protein, was cloned into the shuttle vector pKT240 and recovered as the recombinant pKG201 with a 2.3 kb insert from the CAM plasmid in the PstI site. The gene product is expressed constitutively in P. putida and in E. coli whereas the inverted insert clone lacks expression, indicating absence of an insert promoter. PMID- 2992469 TI - Cloning and sequencing of a full length cDNA corresponding to human cellular retinol-binding protein. AB - We have isolated and sequenced a cDNA clone corresponding to the human cellular retinol-binding protein (CRBP). The deduced amino acid sequence, which encompasses 134 amino acid residues, shows significant homology with several low molecular weight proteins which bind hydrophobic ligands. No homology to the plasma retinol-binding protein was observed. Southern and Northern blot analyses suggest that the CRBP gene is present in a single copy in the haploid genome and that it is transcribed in a single mRNA species. PMID- 2992470 TI - Catabolite inactivation of fructose 1,6-bisphosphatase and cytoplasmic malate dehydrogenase in yeast. AB - Catabolite inactivation of fructose 1,6-bisphosphatase and cytoplasmic malate dehydrogenase was studied using the protease-deficient and vacuole-defective yeast strain pep4-3. The catabolite inactivation of fructose 1,6-bisphosphatase in pep4-3 was found to have a normal first inactivation step but with a defective second proteolytic step. In contrast, catabolite inactivation of cytoplasmic malate dehydrogenase was normal in pep4-3. These results suggest that the proteolytic pathways utilized in the hydrolysis of the two enzymes may be different and that proteolysis of fructose 1,6-bisphosphatase may require functional vacuoles while proteolysis of cytoplasmic malate dehydrogenase may not. PMID- 2992471 TI - Purification and biochemical characterization of alpha 2-adrenergic receptor from the rat adrenocortical carcinoma. AB - The alpha 2-adrenergic receptor was purified from rat adrenocortical carcinoma 494 by an affinity chromatographic step using a novel para-aminoclonidine sepharose resin followed by a gel-permeation high performance liquid chromatographic step. The iodinated receptor protein was homogeneous as evidenced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by high performance liquid chromatography. Both SDS-PAGE and high performance liquid chromatographic studies revealed that Mr of the protein was 64,000, suggesting the monomeric nature of the receptor protein. The purified protein showed the typical binding characteristics of alpha 2-adrenergic receptor. PMID- 2992472 TI - Formation of N tau-ribosylhistidine, a novel histidine derivative found in the urine in histidinemia, from histidine and NAD(P)+ catalyzed by an NAD(P)+ glycohydrolase system. AB - The formation of N tau-ribosylhistidine (His-R), a novel histidine derivative found in the urine of histidinemic patients, was studied. A most possible synthetic pathway catalyzed by imidazole acetic acid (ImAA) phosphoribosyltransferase was not substantiated, because p.o. administration to humans and rats of aspirin, an inhibitor of the enzyme, did not change the urinary excretion of His-R, whereas aspirin decreased the excretion of ImAA-R with concomitant increase in that of ImAA. His-R was produced on incubation of a rat liver homogenate or its membrane fraction with histidine, NAD(P)+ and MgCl2, but not with only histidine or NAD(P)+. Nicotinamide inhibited the formation of His-R. Thus the enzymes responsible for the formation of His-R were suggested to be NAD(P)+ nucleosidase, nucleotide pyrophosphatase and 5'-nucleotidase. PMID- 2992473 TI - Is hydroxyl radical generated by the Fenton reaction in vivo? AB - Yeast mutants deficient in activities of cytosolic superoxide dismutase and catalase A and T were exposed to four different kinds of oxygen stress. The response of the cells contradicts suggestions, that hydroxyl radical is formed in vivo through the Fenton reaction. The results suggest that superoxide radicals are directly responsible for cytotoxic effects of oxygen. PMID- 2992474 TI - Mg2+ efflux is accomplished by an amiloride-sensitive Na+/Mg2+ antiport. AB - Mg2+ efflux from Mg2+-preloaded chicken erythrocytes is caused by an electroneutral Na+/Mg2+ antiport. It depends specifically on extracellular Na+, according to Michaelis-Menten kinetics (Km = 25 mM), and is reversibly noncompetitively inhibited by amiloride (Ki = 0.59 mM). In contrast to Na+/H+ antiport, Li+, Ca2+ and N-ethylmaleimide do not interfere with Na+/Mg2+ antiport. The Na+/Mg2+ antiport is driven by the intracellular/extracellular Mg2+ gradient. PMID- 2992475 TI - Regulatory subunit of cAMP-dependent protein kinase inhibits phosphoprotein phosphatase. AB - The activity of a purified high molecular weight phosphoprotein phosphatase was inhibited by purified type II cAMP-dependent protein kinase. This effect required cAMP and was obtained in the absence of ATP. The isolated type II regulatory subunits (R-subunits) from several species also inhibited the phosphatase activity in both crude extracts and purified preparations. Half maximal inhibition was observed at 0.06-0.25 microM, well within the physiological range of R-subunit concentrations. The inhibitory potency of R-subunit was greater using the thiophosphorylated form. Limited trypsinization of the R-subunit abolished the inhibitory activity. The C-subunit released the bound cAMP when combined with R-subunit, but the phosphatase did not, implying that the inhibited species is a R.cAMP-phosphatase complex. The results suggest that the R-subunit might have at least one physiological role in addition to inhibition of the C subunit, i.e., inhibition of phosphatase. The latter would occur only when cAMP is elevated. PMID- 2992476 TI - Solubilization and molecular weight estimation of atrial natriuretic factor receptor from bovine adrenal cortex. AB - Receptors for atrial natriuretic factor (ANF) have been solubilized with 3-[(3 cholamidopropyl)dimethylammonio]-1-propanesulfonate from bovine adrenal cortex and characterized. The detergent extract retained specific high-affinity binding sites for 125I-ANF. Scatchard analysis of the equilibrium binding data revealed a single class of binding site with a K-d of 1.8 nM and a maximum binding capacity of 2.5 pmol/mg of protein. The size of the 125I-ANF X receptor complexes was estimated to be 140,000 daltons by gel filtration on TSK gel G3000SW. Affinity labeling followed by electrophoresis under nonreducing conditions and autoradiography also revealed a single band of a similar size (Mr = 130,000); this band, however, migrated as a Mr = 70,000 species under reducing electrophoretic conditions. These results indicate that the ANF receptor, having a Mr of 130,000 - 140,000, is composed of disulfide-linked subunits and the ANF binding site is located on the 70-kDa component. PMID- 2992477 TI - Regulation of type IV collagen degrading enzyme by cortisol during human skin fibroblast growth. AB - The effects of cortisol on type IV collagenolytic enzyme activity were studied in human skin fibroblast cultures. Cortisol reduced the production of the enzyme, both in normally dividing fibroblasts, and also when the secretion was induced by the administration of the tumor promoter 12-O-tetradecanoyl phorbol 13-acetate (TPA) to the subconfluent culture. TPA decreased the glucocorticoid receptor activity and augmented the cGMP content of subconfluent fibroblasts. Type IV collagenolytic activity appeared to be correlated positively with the cGMP/cAMP ratio, and negatively with the glucocorticoid receptor activity, and the activity was increased by the addition of dibutyryl cGMP. This suggests that this enzyme may at least partly be under glucocorticoid control during fibroblast growth, and that TPA may to some degree act via the inactivation of glucocorticoid receptors. PMID- 2992478 TI - Activation of calcium mobilization and calcium influx by alpha 1-adrenergic receptors in a smooth muscle cell line. AB - Alpha 1-adrenergic receptor (alpha 1R) mediated increases in the cytosolic levels of free Ca+2 and the inositol phosphates were measured in a smooth muscle cell line, DDT1. Norepinephrine (NE) stimulated a rapid increase in cytosolic Ca+2 by two distinct components: 1) release of Ca+2 from intracellular sites (mobilization), and 2) influx of extracellular Ca+2. The mobilization component was not affected by removal of extracellular Ca+2 or addition of La+3 or Co+2 to the buffer. The influx component was abolished by EGTA, La+3, or Co+2, but was not affected by the voltage-operated Ca+2 channel blockers diltiazem or nifedipine. Depolarization of DDT1 cells with 100 mM KCl or with gramicidin did not induce Ca+2 influx. NE also increased inositol trisphosphate to 78% over basal levels within 1 minute. These results suggest that alpha 1R on DDT1 cells are coupled to both the mobilization of intracellular Ca+2 and to receptor operated Ca+2 channels in the plasma membrane, and that polyphosphoinositide hydrolysis may play a role in these phenomena. PMID- 2992480 TI - Unique inhibitory profile of platelet activating factor induced calcium mobilization, polyphosphoinositide turnover and granule enzyme secretion in rabbit neutrophils towards pertussis toxin and phorbol ester. AB - Platelet activating factor has been found to increase the intracellular level of free calcium (as monitored by the fluorescent calcium indicator quin-2) and to stimulate the turnover of the polyphosphoinositides in rabbit neutrophils. Calcium mobilization induced by platelet activating factor, in contrast to previous reports with chemotactic factors, is unaffected by pertussis toxin; on the other hand, stimulated polyphosphoinositol hydrolysis and granule enzyme secretion are potently antagonized under the same conditions. The calcium, as well as the secretory responses to the lipid mediator are largely dependent on the presence of extracellular calcium. Internal contributions to the quin-2 signal are only detectable at relatively high concentrations of platelet activating factor. Calcium mobilization and secretion stimulated by platelet activating factor are inhibited following a short incubation with phorbol 12 myristate 13-acetate. These results are discussed in terms of the possibility that platelet activating factor activates neutrophils via dual pathways, the first involving direct interaction with phorbol ester inhibitable calcium channels and the other the stimulation in a manner dependent on a guanine nucleotide binding protein of the phospholipase C specific for polyphosphoinositides. PMID- 2992479 TI - Stimulation of Na+/H+ antiport activity by epidermal growth factor and insulin occurs without activation of protein kinase C. AB - Addition of phorbol 12, 13 dibutyrate (PBt2) or a combination of epidermal growth factor (EGF) and insulin to quiescent cultures of Swiss 3T3 cells increased intracellular pH in a Na+-dependent fashion. In contrast to PBt2, EGF plus insulin failed to stimulate protein kinase C and elicited the ionic response in cells lacking this enzyme. We suggest that the stimulation of the Na+/H+ antiport in Swiss 3T3 cells is mediated by at least two separate pathways, only one of which is dependent upon the activation of protein kinase C. PMID- 2992481 TI - Spermine induced phosphorylation of a 42 kD/pI 5.9 nuclear protein in different human tumor cell lines. AB - In vitro phosphorylation of 5 M urea extracts from nuclei obtained from different human tumor cell lines leads to incorporation of phosphate from 32P-gamma-ATP in more than 20 polypeptides with an acidic pI. Whereas heparin at a concentration of 1 microgram had no effect on the phosphorylation pattern, spermine stimulated the total phosphorylation up to twofold. Furthermore, in the presence of this polyamine, the two-dimensional polyacrylamide gel revealed an additional phosphoprotein with an apparent pI of 5.9 and a relative molecular mass of 42 000. Phosphoamino acid analysis of the most prominent phosphoproteins showed serine and threonine as phosphoacceptors. PMID- 2992482 TI - Teleocidin and phorbol ester tumor promoters exert similar mitogenic effects on human lymphocytes. AB - The tumor promoter 12-0-tetradecanoyl-phorbol-13-acetate (TPA) affects a wide variety of cellular functions via its binding to protein kinase C (PKC). The TPA molecule contains a diacylglycerol (DAG)-like structure, which may explain its ability to mimic DAG in PKC activation. Teleocidin (TCD) is a different tumor promoter which can compete with TPA in binding to its cell surface receptors even though structurally unrelated to TPA or DAG. Since TCD may use an additional receptor system and/or be distinguished from TPA in its effect on cells, we compared the effects of TPA and TCD on human peripheral blood lymphocytes (PBL). Both tumor promoters preferentially enhanced cell proliferation of sheep erythrocyte-rosetted lymphocytes, which were enriched for T cells. Additionally, TPA and TCD both induced a high density of cell surface receptors for interleukin 2 (IL2) and transferrin, but not synthesis or production of IL2. However, either of the tumor promoters synergized with T cell mitogens to induce high level IL2 production by PBL. In dose response and kinetic studies, matching concentrations of TPA and TCD induced similar effects in PBL. The results thus demonstrate that TPA and TCD are alike in mitogenic capacity, and suggest that structural similarity between the tumor promoter and DAG, the physiological activator of PKC, is not an essential property for promoting tumors or affecting a wide variety of cellular functions. PMID- 2992483 TI - Homology of ATP binding sites from Ca2+ and (Na,K)-ATPases: comparison of the amino acid sequences of fluorescein isothiocyanate labeled peptides. AB - Ca2+ and (Na,K)-stimulated ATPases from various species and tissues were labeled with fluorescein isothiocyanate (FITC). Labeled peptides were solubilized by tryptic digestion and purified by reverse phase high pressure liquid chromatography. The amino acid sequences of the labeled peptides reveal considerable homology between sarcoplasmic reticulum Ca2+-ATPases from various sources. These Ca2+-ATPases also contain a region of homology with all other ATPases thus far sequenced. A difference was demonstrated between dog skeletal and cardiac Ca2+-ATPases. These results demonstrate homology of the putative ATP binding site of ATPases, which extends over tissue, species, and cation specificity, including the completely conserved amino acid sequence: lys-gly-ala pro-glu. PMID- 2992485 TI - Expression of the c-sis gene and secretion of a platelet-derived growth factor like protein by simian virus 40-transformed BHK cells. AB - SV40-transformed BHK cells were shown to express two transcripts, of 3.5 kb and 2.0 kb, that hybridised to a human c-sis probe. Antibodies directed against human PDGF specifically recognized a 31 kDa protein in SV40/BHK cell conditioned medium, which upon reduction was split into 16 kDa species. Unfractionated conditioned medium and one of two growth factors isolated from SV40/BHK cells competed with 125I-PDGF for binding to its receptor. The present communication thus provides compelling evidence that an SV40/BHK cell-derived growth factor is a hamster equivalent to human PDGF. PMID- 2992484 TI - Effect of doxorubicin-enhanced hydrogen peroxide and hydroxyl radical formation on calcium sequestration by cardiac sarcoplasmic reticulum. AB - This study investigated the effect of doxorubicin-related oxygen radical formation on Ca2+ uptake by rat heart sarcoplasmic reticulum vesicles. Enzymatic activation of doxorubicin by cardiac NADH dehydrogenase produced a dose-related inhibition of Ca2+ uptake that was enzyme- and cofactor-dependent and that was inhibited by catalase, various hydroxyl radical scavengers, and the iron chelator deferoxamine. Furthermore, inhibition of Ca2+ uptake paralleled the production of the hydroxyl radical by NADH dehydrogenase after doxorubicin treatment. These results suggest that doxorubicin-stimulated reactive oxygen metabolism can alter Ca2+ transport by cardiac sarcoplasmic reticulum and may represent one pathway involved in the cardiac toxicity of this potent antineoplastic agent. PMID- 2992486 TI - High level human interleukin 1 production by a hepatoma cell line. AB - The human hepatic adenocarcinoma cell line, SK-hep-1, was found to constitutively produce Interleukin 1. Addition of the ionophore A23187 and lipopolysaccharide resulted in a 30-fold enhancement in the release of biological activity. Serum supplementation did not affect the level of production. Interleukin 1 from these cells had a molecular weight of 10-20,000 daltons on gel exclusion chromatography. Polyadenylated RNA, when fractionated on sucrose density gradients and injected into Xenopus laevis oocytes, produced high levels of biological activity in the 14-16s region. An oligonucleotide probe, complementary to the coding sequence of the Interleukin 1 cDNA isolated from human monocytes, hybridized specifically to this part of the gradient. These results demonstrate that SK-hep-1 cells are a valuable source of material for studying the polypeptide and messenger RNA of Interleukin 1. PMID- 2992487 TI - Evidence for the phosphorylation of the type II insulin-like growth factor receptor in cultured cells. AB - The ATP pools of monolayer cultures of rat embryo fibroblasts and rat liver cells (BRL-3A2) were labeled with [32P]H3PO4. The type II insulin-like growth factor (IGF) receptor was purified by affinity chromatography on wheat germ lectin Sepharose and IGF-II-Sepharose columns. A phosphorylated species having the expected size of the type II receptor (Mr = 220,000 without reduction, Mr = 260,000 with reduction) was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. IGF-II stimulated phosphorylation of the type II receptor in BRL-3A2 rat liver cells. Lability of the receptor phosphate bonds to alkaline pH suggests that the bulk of phosphorylation was occurring on serine residues. PMID- 2992488 TI - The presence of two mitochondrial DNA topoisomerases in human acute leukemia cells. AB - We have undertaken a study of DNA topoisomerases in mitochondria from human acute leukemia cells. Two activities have been detected in these organelles. One of the enzymes is presumably a type II topoisomerase, i.e., in ATP-dependent reactions it can catenate closed circular plasmid DNA, and decatenate closed circular kinetoplast DNA. A second topoisomerase is presumably a type I enzyme since, it can relax positive as well as negative supercoils in an ATP-independent reaction, it is unable to catenate plasmid DNA or decatenate kinetoplast DNA, and it is inhibited, rather than stimulated, by ATP. PMID- 2992489 TI - Biosynthesis of a sulfonolipid in gliding bacteria. AB - Gliding bacteria of the genus Cytophaga synthesize sulfonolipids (1,2) that contain capnine (1-deoxy-15-methylhexadecasphinganine-1-sulfonic acid). Studies of the incorporation of radiolabeled compounds by C. johnsonae show that cysteate is utilized preferentially to both cystine and inorganic sulfate as a precursor of capnine sulfur and to both cystine and serine as a precursor of carbons 1 and 2 of capnine. The results are consistent with a pathway in which capnine is formed by condensation of cysteate with a fatty acyl CoA. Cystine, added as the sole sulfur source in the presence of glucose, provides the sulfur but not the carbon for capnine. Hence, these cells form cysteate not by direct oxidation of cystine (or cysteine), but by transfer of its sulfur to a different carbon compound. PMID- 2992490 TI - Involvement of adenosine kinase in the phosphorylation of formycin B in CHO cells. AB - In Chinese hamster ovary cells, [3H]formycin B is metabolized into formycin B-5' monophosphate, formycin A-5'-monophosphate and higher phosphorylated derivatives of formycin A which are incorporated into RNA. Mutants of CHO cells independently selected for resistance to various adenosine analogs viz. toyocamycin, tubercidin, 6-methylmercaptopurine riboside, which contain no detectable activity of adenosine kinase (AK) in cell extracts, all exhibited between 2- to 3-fold increased resistance to formycin B. Formycin B-resistant mutants of CHO cells are also affected in AK, as indicated by the absence of AK activity in cell extracts. Both types of AK- mutants showed reduced uptake and phosphorylation of [3H]formycin B in comparison to the parental (AK+) cells. In addition, toxicity of formycin B towards CHO cells was reduced in presence of adenosine in a concentration dependent manner. These observations strongly indicate that in CHO cells, formycin B is phosphorylated via AK and that like other nucleoside analogs its phosphorylation may be essential for the drugs cellular toxicity. PMID- 2992491 TI - Conversion and inactivation of opioid peptides by rabbit brain endo oligopeptidase A. AB - The conversion of BAM-12P to Met-enkephalin and the hydrolysis of the Phe-Met and Phe-Leu bonds of met-enkephalin-Arg-Phe and Leu-enkephalin-Arg-Arg, respectively, by rabbit brain endo-oligopeptidase A were demonstrated. Peptide fragments were isolated by high performance liquid chromatography and identified by amino acid analysis. BAM 22P was not hydrolysed by the enzyme. The concentration dependent inhibition of BAM-12P conversion into Met-enkephalin by bradykinin and vice-versa provided additional evidence that endo-oligopeptidase A cleaves both the Phe5 Ser6 bond in bradykinin and the Met5-Arg6 bond of BAM-12P. PMID- 2992492 TI - A regulatory protein for orthophosphate-regulated cyclic phosphodiesterase in Neurospora crassa. AB - A regulatory protein for orthophosphate-regulated cyclic phosphodiesterase (cPDase) was detected in mycelial extracts of Neurospora crassa. The protein, designated neucrassin, was precipitated by ammonium sulfate between 60 to 100% saturation, and fractionated by gel filtration through a TSK-gel column. The molecular weight was estimated to be 65,000. Neucrassin inhibited the hydrolyzing activity of cPDase for cyclic 3',5'-AMP in the presence of MnCl2 in a noncompetitive manner, whereas it stimulated the activity for cyclic 2',3'-AMP over a wide ranges of pH's, between 2.8 to 6.8. PMID- 2992493 TI - "Differential displacement"--a simple method to study receptors for 1,25 dihydroxyvitamin D3 in the presence of contaminating serum vitamin D binding protein. AB - This report describes a simple procedure to quantitate receptors for 1,25 dihydroxyvitamin D3 in the presence of contaminating serum vitamin D binding protein. The method ("differential displacement") takes advantage of the greatly different rates of dissociation of 1,25-dihydroxyvitamin D3 from the serum vitamin D binding protein (t 1/2 less than 5 min) and from the receptor (t 1/2 greater than 120 h) at 0 degrees C. The principle of "differential displacement" can be used for other steroid receptors as well, and in combination with a variety of different binding assays, provided they are performed under conditions where the dissociation of the steroid from the receptor is slow. PMID- 2992495 TI - Electrochemistry of norcocaine nitroxide and related compounds: implications for cocaine hepatotoxicity. AB - Norcocaine nitroxide, a free radical metabolite of cocaine, displays a reversible one-electron cyclic voltammogram which is abolished by the addition of reduced glutathione. The corresponding nitrosonium ion was synthesized. It showed the same electrochemical characteristics as the nitroxide. The spin label 4-hydroxy 2,2,6,6-tetramethyl piperidine-1-oxyl (TEMPOL) and its nitrosonium ion behaved like morcocaine nitroxide and its nitrosonium ion. The nitrosonium ion of TEMPOL caused hemolysis of red blood cells, but TEMPOL did not. These observations suggest that the highly reactive nitrosonium ion may be involved in the production of cocaine-induced hepatic necrosis in mice. PMID- 2992496 TI - Pertussis toxin reversal of the antilipolytic action of insulin in rat adipocytes in the presence of forskolin does not involve cyclic AMP. AB - Insulin inhibition of lipolysis in the presence of forskolin was reversed by a four hour exposure of adipocytes to pertussis toxin. In contrast, the antilipolytic action of insulin against lipolysis due to theophylline was unaffected by pertussis toxin as was the ability of insulin to lower cyclic AMP in the presence of either forskolin or theophylline. The stimulation of adenylate cyclase by norepinephrine in crude plasma membranes obtained from rat adipocytes was inhibited by N6-(Phenylisopropyl)adenosine (PIA) and abolished by pretreating rat adipocytes with pertussis toxin. The stimulation of glucose metabolism by insulin was not altered by pertussis toxin pretreatment of rat adipocytes. These findings suggest that pertussis toxin selectively abolishes the antilipolytic effect of insulin in the presence of forskolin through a cyclic AMP independent mechanism. PMID- 2992497 TI - Lack of effect of synthetic atrial natriuretic factor on rubidium uptake by human erythrocytes. AB - The effect of synthetic atrial natriuretic factor on the ouabain-sensitive and the furosemide-sensitive rubidium uptake by human erythrocytes has been studied. This peptide with potent diuretic and natriuretic effects did not affect any rubidium uptake system at concentrations of 10(-7) and 10(-9) M. These results do not support that the natriuretic effect is based on the inhibition of active transport systems in the renal tubules. PMID- 2992494 TI - Effect of epidermal growth factor on synthesis of prostaglandins and cyclic AMP by embryonic palate mesenchymal cells. AB - The influence of epidermal growth factor (EGF) on the ability of murine embryonic palate mesenchymal (MEPM) cells to be stimulated to synthesize cAMP and prostaglandins was investigated. Preincubation of MEPM cells with EGF enhanced, in a dose-dependent fashion, (1) the responsiveness of MEPM cells to prostaglandin E1-induced elevation of intracellular levels of cAMP, and (2) the responsiveness of cells to calcium ionophore (A23187) and melittin-induced synthesis of prostaglandins E2 and F2 alpha. Hormonal responsiveness of MEPM cells to EGF, prostaglandins and cAMP has been implicated as being involved in controlling various aspects of normal oro-facial development. We show here that EGF can potentiate hormonal responsiveness of these cells and thus allows consideration of EGF as a factor which may modulate hormonally regulated craniofacial growth and differentiation. PMID- 2992498 TI - A novel tumor-associated molecular species of 5'-nucleotide phosphodiesterase. AB - 5'-Nucleotide phosphodiesterase (5'-NPDase) was partially purified from plasma membranes of murine lymphoma L5178Y. The enzyme was inactivated by N ethylmaleimide and Zn2+, but stabilized by dithiothreitol, suggesting that it is an SH enzyme. The enzyme, Km 1.54 mM, pI 5.8 and MW 23k, differs from liver 5' NPDase in MW, Km and sensitivity to some inhibitors. On the contrary, 5'-NPDase, derived from normal mouse organs, is similar to the liver enzyme. The results suggest that tumor cells possess a novel molecular species of 5'-NPDase. PMID- 2992499 TI - The energetics of the injection process of bacteriophage lambda DNA and the role of the ptsM/pel-encoded protein. AB - We have examined the nature of the role played in the process of phage lambda DNA injection by the bacterial protein coded by the ptsM/pel gene. Neither the specific inhibition of the activity of the PtsM protein, nor the addition of inhibitors of phosphotransferase system modified the efficiency of lambda DNA penetration. Thus, the PtsM/Pel protein does not seem to play a role through its transport function, although we have confirmed that it must be present for a successful lambda DNA injection. Moreover, the presence of various metabolic inhibitors (uncouplers, cyanide, arsenate) separately or together, or even harsher methods of energy depletion did not prevent lambda DNA penetration, suggesting that DNA is entering the cell cytoplasm by diffusion. PMID- 2992500 TI - Preliminary 1H-NMR studies of the interaction between cytochrome c3 and ferredoxin I from Desulfovibrio desulfuricans Norway. AB - The complex formation of two electron transfer proteins, cytochrome c3 and ferredoxin I from Desulfovibrio desulfuricans Norway, has been shown by 1H-NMR spectroscopy. Presence of ferredoxin I produces ferricytochrome c3 1H-NMR spectrum modifications. The chemical shift of perturbated heme methyl resonances has been used to determine the stoichiometry of the complex. At pH 7.6 and 20 degrees C, the two proteins were found to form a complex 1:1 with an association constant, KA, of 10(4) M-1. Two of the four hemes are affected by presence of ferredoxin I and may be involved in the electron transfer sites. The heme methyl resonances are average resonances of free and bound cytochrome c3 resonances, indicating a fast exchange process on the NMR time scale. PMID- 2992501 TI - Inositol 1,4,5-trisphosphate induces cyclic GMP formation in Dictyostelium discoideum. AB - Chemotactic signalling in the cellular slime mould Dictyostelium discoideum employs signalling molecules such as folate and cyclic AMP. These bind to specific cell surface receptors and rapidly trigger internal responses that induce chemotactic movement of the amoebae. Previous studies have shown that actin is polymerised within 3-5 sec of cyclic AMP or folate binding and that a peak of cyclic GMP is formed within 9-12 sec. Release of Ca2+ from intracellular stores has been implicated as a secondary messenger. Here we present evidence that D-myo-inositol 1,4,5-trisphosphate, when added to permeabilized amoebae of Dictyostelium, can mimic the action of chemoattractants on normal intact amoebae in inducing cyclic GMP formation. Our data suggest that IP3, which is known to act as an intermediary messenger between cell surface hormone receptors and release of Ca2+ from internal stores in mammalian cells, functions in a similar capacity during chemotaxis of this primitive eukaryote. PMID- 2992502 TI - Atrial natriuretic factor increases cyclic GMP and inhibits cyclic AMP in rat renal papillary collecting tubule cells in culture. AB - The present study was undertaken to determine whether human atrial natriuretic factor (hANF) produces guanosine-3', 5'-monophosphate (cGMP) and alters arginine vasopressin (AVP)- and forskolin (F)- induced adenosine-3', 5'-monophosphate (cAMP) production in the cultured rat renal papillary collecting tubule cells. hANF increased cellular cGMP levels in a dose dependent manner. AVP and F, however, did not affect cGMP production. hANF significantly inhibited AVP- and F stimulated cAMP levels, but hANF by itself did not affect cellular cAMP production. Since F activates adenylate cyclase at a step of catalytic unit and the cellular action of AVP to activate adenylate cyclase is mediated through receptor-catalytic units, the present results indicate that hANF may directly inhibit the AVP- and F-stimulated adenylate cyclase in renal papillary collecting tubules. PMID- 2992503 TI - Two classes of cAMP analogs synergistically inhibit p21 ras protein synthesis and phenotypic transformation of NIH/3T3 cells transfected with Ha-MuSV DNA. AB - Factors that control cellular proliferation might do so by regulating quantitative expression of viral or cellular oncogenes. Since the growth regulatory effect of cAMP is well-known, the effect of cAMP on ras gene expression was examined on Ha-MuSV-transformed 13-3B-4 cells (NIH-3T3) grown in chemically defined serum-free medium. Treatment of cells with two classes of cAMP analogs which are selective for the two different cAMP-binding sites of type II protein kinase, in combination, synergistically inhibited both p21 ras protein synthesis and phenotypic transformation. The inhibition was also demonstrated with these analogs singly but at higher concentrations. The decrease in p21 synthesis was inversely correlated with an increase in the RII cAMP receptor protein, the regulatory subunit of type II protein kinase. These results suggest a role for cAMP and its receptor protein in the regulation of v-rasH oncogene expression. PMID- 2992504 TI - Luteinizing hormone-releasing hormone enhances polyphosphoinositide breakdown in rat granulosa cells. AB - A 2-min addition of LHRH to [3H]inositol-prelabeled rat granulosa cells in primary culture evoked significant increases in the accumulation of [3H]inositol phosphates, i.e. radiolabeled inositol monophosphate (IP), inositol diphosphate (IP2), and inositol triphosphate (IP3) levels increased to 210, 590 and 520%, respectively, when compared to control cultures. By contrast, addition of FSH failed to elicit such a response. The effect of LHRH was completely blocked by the concomitant presence of a specific LHRH antagonist. LHRH evoked increase in [3H]IP3 and [3H]IP2 accumulation as early as 30 sec, while the increase in [3H]IP became significant at 2 min. These data support the hypothesis that polyphosphoinositide breakdown may be an early step in the intracellular signal mechanism which mediates the action of LHRH. PMID- 2992505 TI - Properties of interleukin-1 as a complete secretagogue for human neutrophils. AB - Human monocyte-derived Interleukin-1 (IL-1) stimulated a concentration-dependent extracellular release of azurophil (myeloperoxidase) and specific (vitamin B12 binding protein) granule constituents from cytochalasin B-treated human neutrophils. The serine protease inhibitors, L-1-tosylamide-2-phenylethyl chloromethyl ketone (TPCK) and N-alpha-p-tosyl-L-lysine-chloromethyl ketone (TPCK) as well as an inhibitor of thiol protease activity, p hydroxymercuribenzoate (PHMB), suppressed granule enzyme release from neutrophils activated with IL-1. Cycloheximide, an inhibitor of protein synthesis, had no effect on IL-1-induced neutrophil degranulation. Neutrophils pretreated with IL-1 were rendered unresponsive to subsequent exposure to this stimulus. IL-1-elicited granule exocytosis appears to be stimulus specific in that N-formyl-methionyl leucyl-phenylalanine (FMLP), 1-0-hexadecyl/octadecyl-2-0-acetyl-sn-glyceryl-3 phosphorycholine (AGEPC), and 5(S),12(R)-dihydroxy-6,14-cis-8,10-trans eicosatetraenoic acid (LTB4) were capable of eliciting a secretory response from IL-1-pretreated cells. PMID- 2992506 TI - Synthetic analogs of the active site of cytochrome P-450cam characterization of thiolpeptide fragment-hemin complexes by optical and ESR spectrometries. AB - Thiol-containing penta (Leu-Ala-Cys-Ser-Leu) and nona (Leu-Ala-Cys-Ser-Leu-Ile Glu-Ser-Leu) peptides corresponding to residues 132-136 and 132-140, respectively, of apo-P450 from Psuedomonas putida were synthesized to examine heme-binding by the enzymes in the oxidized (ferric) form. The peptide-hemin complexes prepared in solution were characterized by their optical and ESR spectra. In these complexes without nitrogenous ligands, no ferric complexes in the low-spin state were observed, suggesting that simultaneous axial coordination of Cys-134 (thiolate) and Ser-139 (hydroxyl) of apo-P450cam to the heme is unlikely to occur. In the presence of nitrogenous ligands, such as py, Im and MeIm, the resulting complexes were good models of apo-P450cam-nitrogenous ligand adducts in the low-spin ferric state retaining a thiolate-Fe(III)-nitrogen axial coordination mode, as judged by their spectral pattern as well as by crystal field analysis of ESR g-values. PMID- 2992507 TI - Isolation and characterisation of a variant allele of the gene for human apolipoprotein E. AB - Size-selected human DNA fragments enriched in the Apolipoprotein E (ApoE) gene sequence were cloned from an individual of known ApoE phenotype, E2/E2. The clone bank was screened using a human cDNA clone for the ApoE locus (1), and a single genomic clone was isolated. Sequence data obtained from appropriate subcloned fragments confirmed that the codon for Arg-158 (CGC) in the E3 allele is altered to the codon for Cys (TGC) in the E2 allele. Hybridisation data indicated the presence of at least one intron in the ApoE gene, consistent with the structure of an independently isolated human ApoE4 allele (2). PMID- 2992508 TI - Proton NMR study of the cytochrome c:flavodoxin electron transfer complex. AB - The effects of complex formation with flavodoxin on the proton NMR spectrum of cytochrome c are to change the resonance frequencies and to increase the bandwidths of most of the low and high field heme, Met-80, and His-18 protons. These effects are, in general, more pronounced than has been reported for other cytochrome c complexes. The degree of line broadening for many heme related resonances suggests that complex formation induces changes in the cytochrome structure. These results provide the first spectroscopic evidence which corroborates the proposed model for the cytochrome c: flavodoxin complex (1-3). PMID- 2992509 TI - Diverse involvements of Ni protein in superoxide anion production in polymorphonuclear leukocytes depending on the type of membrane stimulants. AB - Effects of islet-activating protein (IAP) were examined to assess the involvement of the guanine nucleotide-binding regulatory protein responsible for inhibition of adenylate cyclase system (Ni protein) in the superoxide anion (O-2) production in polymorphonuclear leukocytes (PMNL) stimulated with various agents. N-Formyl methionyl-leucyl-phenylalanine (FMLP)-stimulated O-2 production was inhibited by the pretreatment with IAP. O-2 production induced by each of phorbol myristate acetate, concanavalin A, and A23187, however, was rather resistant to the pretreatment with IAP. This observation indicates that the Ni protein does not involve in the common pathway for the O-2 production. in PMNL, and the involvement is rather specific for the FMLP-induced production. O-2 production in PMNL stimulated with various membrane perturbing agents was also diverse in the requirement of extracellular Ca2+. PMID- 2992510 TI - Inhibition by vanadyl of adenylate cyclase activation reactions in rat adrenal membranes. AB - In rat adrenal dispersed cells, both vanadyl and vanadate inhibited ACTH stimulated steroidogenesis and the formation of cAMP, whereas these compounds did not inhibit the cAMP-dependent steroidogenesis. Then, the membrane fraction was prepared and activated by various secretagogues including ACTH, Gpp(NH)p, GTP gamma S, and forskolin. The cyclase activity was inhibited by vanadyl but not by vanadate in the presence of these stimulators. Based on these results, we conclude that cationic vanadyl acts against a metal-requiring step in the adenylate cyclase system containing G-protein and the catalytic subunit. In addition, we believe that vanadyl is a useful tool to investigate adenylate cyclase systems. PMID- 2992511 TI - Phosphorylation of neurofilament proteins by endogenous calcium/calmodulin dependent protein kinase. AB - A protein fraction containing neurofilaments was prepared from rat brain cytosol by differential centrifugation and gel filtration chromatography. These preparations were enriched for a calcium/calmodulin-dependent kinase activity that phosphorylated endogenous neurofilament proteins. The enzyme incorporated approximately 1 mol PO4/mol of each neurofilament triplet polypeptide. These data suggest that a calmodulin-dependent kinase may mediate some of the effects of calcium on cytoskeletal function by phosphorylation of neurofilament proteins. PMID- 2992513 TI - Photoaffinity labeling of atrial natriuretic factor receptor in bovine and rat adrenal cortical membranes. AB - Radioiodinated synthetic atrial natriuretic factor (ANF) bound to a single class of high affinity binding sites in the plasma membrane from bovine adrenal cortex with a KD of 7.4 X 10(-10) M. The binding affinities of related peptides showed close parallelism to their potencies in natriuretic and vasorelaxant activities. Incubation of adrenal membranes with radioiodinated 4-azidobenzoyl ANF or a similar derivative of its analogue followed by photolysis resulted in specific radiolabeling of a protein band in SDS gel electrophoresis with an apparent Mr of 124,000 in bovine or Mr of 126,000 in rat, which was abolished by inclusion of unmodified ANF in the incubation. Prevention of the labeling was dependent on the concentration of ANF and was not observed with atriopeptin I or with unrelated peptides, angiotensin II, ACTH or [Arg8] vasopressin. These results indicate specific covalent labeling of ANF-receptor or its subunit by the photoaffinity ligands. PMID- 2992512 TI - Threonine phosphorylation of rat liver glycogen synthase. AB - 32P-labeled glycogen synthase specifically immunoprecipitated from 32P-phosphate incubated rat hepatocytes contains, in addition to [32P] phosphoserine, significant levels of [32P] phosphothreonine (7% of the total [32P] phosphoaminoacids). When the 32P-immunoprecipitate was cleaved with CNBr, the [32P] phosphothreonine was recovered in the large CNBr fragment (CB-2, Mapp 28 Kd). Homogeneous rat liver glycogen synthase was phosphorylated by all the protein kinases able to phosphorylate CB-2 "in vitro" (casein kinases I and II, cAMP-dependent protein kinase and glycogen synthase kinase-3). After analysis of the immunoprecipitated enzyme for phosphoaminoacids, it was observed that only casein kinase II was able to phosphorylate on threonine and 32P-phosphate was only found in CB-2. These results demonstrate that rat liver glycogen synthase is phosphorylated at threonine site(s) contained in CB-2 and strongly indicate that casein kinase II may play a role in the "in vivo" phosphorylation of liver glycogen synthase. This is the first protein kinase reported to phosphorylate threonine residues in liver glycogen synthase. PMID- 2992514 TI - A comparison of the influence of diets high in saturated versus unsaturated fatty acids on lipid composition and glucose-6-phosphatase activity of rat liver microsomes. AB - Glucose-6-phosphatase (G6Pase) activity in microsomes (M-SF) prepared from livers of rats fed a saturated fat-enriched diet has been observed to be 20-30% higher than in microsomes (M-PUF) prepared from rats fed a polyunsaturated fat-enriched diet. This difference was persisted throughout the 49 days experimental period. Neutral lipids of M-PUF contained a significantly higher concentration of 18:2 and 20:4, and a lower concentration of 16:0, 18:0 and 18:1 whereas in phospholipids 18:2 and 20:4 accumulated at the expense of 16:0 as compared to M SF. Total phospholipid content from M-PUF was 30% lower than that from M-SF but the proportional phospholipid composition remained the same. An increase in cholesterol content was observed in M-PUF and this increase was localized in the ester fraction of cholesterol. These observations clearly indicate that manipulations of dietary lipids can influence both the fatty acid profiles and enzyme function of hepatic microsomal membranes. PMID- 2992515 TI - Isolation and partial characterization of BstVI, a thermostable isoschizomer of XhoI. AB - A type II restriction endonuclease, which has been named BstVI, was isolated and partially purified from a spore-forming, gram-positive thermophilic bacilli. On the basis of its digestion patterns on various DNA's, it was concluded that this enzyme is an isoschizomer of XhoI, isolated originally from Xanthomonas holcicola. Besides being highly thermostable, the enzyme is produced in very large amounts by this bacterial strain. A single purification step renders it free of unspecific nucleases and suitable for performing restriction analysis and cloning experiments. PMID- 2992516 TI - Hormonal regulation of fat esterification & oxidation in hepatocytes from fed rats: modulation by 3-mercaptopicolinate. AB - 3-Mercaptopicolinate, an inhibitor of phosphoenolpyruvate carboxykinase (PEPCK), decreased esterification of [1-14C] oleate and [1-14C] myristate in hepatocytes from fed rats. In the absence of 3-mercaptopicolinate, adrenaline, noradrenaline, vasopressin or angiotensin II increased esterification to triacylglycerol of [1 14C] oleate but not [1-14C] myristate. Cyclic AMP decreased esterification of both oleate and myristate. In the presence of 3-mercaptopicolinate, stimulation of oleate esterification by the catecholamines, vasopressin or angiotensin II was increased, and stimulatory effects of these hormones on myristate esterification were observed. Adrenaline, noradrenaline, vasopressin or angiotensin II increased 14CO2 production from both [1-14C] oleate and [1-14C] myristate but the degree of stimulation was similar in the absence or presence of 3-mercaptopicolinate. The results indicate a role for the catecholamines and angiotensin II in the regulation of liver fat metabolism and emphasize the potential importance of changes in activity of PEPCK as determinants of hepatic carbon flux. PMID- 2992517 TI - Chemical mechanisms for photoaffinity labeling of receptors. PMID- 2992518 TI - Identification of specific binding sites for leukotriene C4 in membranes from human lung. AB - Leukotriene C4 (LTC4), one of the major components of the slow-reacting substance of anaphylaxis (SRS-A), is a potent constrictor of bronchial smooth muscle in many species including humans. Here we report the identification and characterization of specific binding sites for LTC4 in membranes from human lung parenchyma. At 4 degrees, 3H-LTC4 binding is specific, saturable (Bmax = 32-41 pmoles/mg prot.), rapid (equilibrium being attained within 15 min), reversible and of high affinity (Kd = 3.6-7 X 10(-8) M). The binding sites are sensitive to heat and probably possess a protein moiety, being inactivated upon trypsinization. CaCl2 affects both the association and the dissociation rate and dose-dependently enhances the binding of 3H-LTC4 at equilibrium; maximal enhancement (4-fold) occurred at 10(-2)M CaCl2. Unlabelled LTC4 is able to complete with 3H-LTC4 for its binding sites with an IC50 of 7.8 X 10(-8) M. The addition of 10(-2) M CaCl2 increases the potency of LTC4 in inhibiting the binding (2.2-fold); both the competition curves are monophasic, indicating the existence of a homogeneous class of binding sites. In the presence of CaCl2, LTD4, LTE4 and the SRS-A antagonist FPL 55712 can inhibit 3H-LTC4 specific binding, being, however, less potent than LTC4 (IC50 S = 2.2 X 10(-6), 2.4 X 10( 5) M, for LTD4, LTE4 and FPL 55712, respectively). FPL 55712 displayed a competitive mechanism; its affinity, however, was lower if absorption to glass was not prevented. The present studies indicate that specific binding sites for 3H-LTC4 exist in human lung parenchyma, and that a receptor-mediated process might be involved in the bronchoconstriction induced by LTC4. PMID- 2992519 TI - The mechanism of inhibition by 2,2'-pyridylisatogen tosylate of NADPH-linked enzyme activities in microsomes isolated from rat liver. AB - Microsomal preparations isolated from rat liver were used to study the action of 2.2'-pyridylisatogen tosylate (PIT) on aniline hydroxylation, cytochrome c reduction and NADPH oxidation. PIT was found to inhibit both the NADPH-dependent (5-100 microM, PIT) and the NADPH-independent (0.05-2.5 mM, PIT) hydroxylation of aniline, but had no significant effect on either the NADPH-dependent oxidation of hexobarbital, or the NADPH-independent hydrolysis of glucose-6-phosphatase. PIT was also found to inhibit cytochrome c reductase competitively (Ki = 35 microM) and to stimulate NADPH oxidation (ED50 = 6.5 microM) PIT and aniline were both found to bind to the microsomal haemoprotein cytochrome P-450 and produce Type II spectral changes. It is proposed that PITs ability to bind to the haemoprotein and its ability to accept electrons from the microsomal NADPH-cytochrome c reductase system leads to the inhibition of aniline hydroxylase activity. PMID- 2992520 TI - Anti-morphine anti-idiotypic antibodies. Opiate receptor binding and isolated tissue responses. AB - Anti-idiotypic antibodies which recognize the opiate receptor were generated in guinea pigs following immunization against purified rabbit anti-morphine antibodies. The anti-idiotypic antibodies produced a concentration-dependent inhibition of [3H]naloxone binding to opiate receptors in a membranous mouse brain preparation. Saturation analysis indicated that the antibodies produced a non-competitive inhibition of naloxone binding. The ability of the antibodies to interact with biological systems was investigated in in vitro systems. In both the isolated guinea pig ileal longitudinal muscle and mouse vas deferens, the antibodies produced a concentration-dependent, opiate agonist-like action. The anti-morphine anti-idiotypic antibodies appear to interact specifically with the opiate receptor and may serve as useful tools in characterization of this receptor system. PMID- 2992521 TI - Factors controlling beta 1-adrenoceptor affinity and selectivity. AB - A membrane preparation of the calf heart left ventricle has been used after identification and characterization, as a source of myocardial beta 1 adrenoceptor for radioligand binding studies. The displacement of specifically bound (-)-[3H]dihydroalprenolol by some beta-adrenoceptor ligands appeared to be pH-dependent, which could be related to the ionization characteristics of the compounds. Among the usually four ionic species of the ligand, present at physiological pH, the cation was shown to govern beta 1-adrenoceptor affinity. Furthermore, quantitative structure affinity relationships for the interaction with beta 1- and beta 2-adrenoceptors were established for the phenoxypropanolamines, a class of beta-adrenoceptor ligands. The N-isopropyl oxypropanolamine side chain itself does not discriminate between beta 1- and beta 2-adrenoceptors, whereas aromatic substitution ortho to the side chain induces some beta 2-selectivity. Selectivity for myocardial beta 1-adrenoceptors is mainly obtained by aromatic substitution para to the side chain. This substitution pattern yields a decrease in beta 2-adrenoceptor affinity, far more pronounced than the decrease in beta 1-adrenoceptor affinity. PMID- 2992522 TI - Evaluation of microsomal pathways of oxidation of alcohols and hydroxyl radical scavenging agents with carbon monoxide and cobalt protoporphyrin IX. AB - Rat liver microsomes catalyze the oxidation of hydroxyl radical scavenging agents by an iron-dependent process, and can oxidize alcohols by pathways dependent on, as well as independent of, .OH. Experiments were carried out to evaluate which microsomal components participate in the production of .OH, and in the two pathways of oxidation of alcohols. Cobalt protoporphyrin IX treatment of rats resulted in a decrease in microsomal oxidation of aminopyrine, .OH scavengers, and alcohols. However, this treatment not only lowered the content of cytochrome P-450, but also decreased the activity of NADPH-cytochrome P-450 reductase. Carbon monoxide, metyrapone and SKF-525A also inhibited the oxidation of aminopyrine but did not affect oxidation of .OH scavengers. Desferrioxamine, a potent iron chelator, inhibited the oxidation of .OH scavengers but not aminopyrine. The oxidation of alcohols was partly sensitive to desferrioxamine and partly sensitive to carbon monoxide, thus showing similarities to the oxidation of .OH scavengers and drugs. These results suggest that the desferrioxamine-sensitive, .OH-dependent pathway of alcohol oxidation is mediated by the reductase, in analogy to results with .OH scavengers, whereas the desferrioxamine-resistant pathway of alcohol oxidation is mediated by cytochrome P-450, in analogy to results with aminopyrine. By the use of desferrioxamine or carbon monoxide, either of the two alcohol-oxidizing pathways can be inhibited independently of each other. PMID- 2992523 TI - Hormonal regulation of heme oxygenase induction in avian hepatocyte culture. AB - The effects of various hormones were examined on the induction of heme oxygenase in monolayer cultures in chick embryo hepatocytes maintained in a chemically defined medium. Addition of insulin to the cultured cells markedly suppressed the activity of basal as well as Co2+-induced heme oxygenase. Treatment of cells with hydrocortisone also suppressed the basal enzyme activity, while the Co2+-induced enzyme activity was enhanced slightly. In contrast, triiodothyronine addition to the culture caused a slight increase of both uninduced and induced levels of the enzyme. This stimulatory effect of triiodothyronine was enhanced significantly by prolonged incubation of cells (48-96 hr) in the serum-free medium. These findings indicate that heme oxygenase synthesis can be substantially altered by changing the hormonal environment of the hepatocytes. Furthermore, the induction of heme oxygenase by Co2+ was inhibited by glucagon, dibutyryl cAMP and theophylline in a dose-dependent manner, suggesting that the enzyme induction may also be controlled by changes in cAMP levels. PMID- 2992524 TI - Protonophoric properties of fluorinated arylalkylsulfonamides. Observations with perfluidone. AB - The acidity and lipophilicity of the fluorinated arylalkylsulphonamides are determined by the nature of the substituents on their aromatic rings. Herbicidal and anti-inflammatory effects of these compounds appear to increase with their lipophilicity. According to Mitchell's chemiosmotic theory, lipophilic weak-acid uncoupling agents act by transporting protons across the inner mitochondrial membrane and thus destroying the proton-electrochemical potential gradient required for ATP synthesis and ion transport. 1:1:1-Trifluoro-N-[2-methyl-4 (phenylsulphonyl) phenyl]methanesulphonamide (Perfluidone), a pre- and post emergence herbicide (at 20 microM concentration), in isolated rat-liver mitochondria caused (1) a 2-fold stimulation of metabolic state-4 respiration, (2) a reduction of respiratory control ratio (RCR) by at least 50%, (3) an enhancement of latent ATPase activity by 40%, (4) a significant passive swelling of mitochondria in 0.15 N NH4Cl(delta A520 = -0.46 +/- 0.003), (5) proton intrusion during state-4 respiration (356 ng H+/min/mg protein; ng H+/min/mg protein with 5 microM perfluidone), and (6) at least 100% stimulation of oligomycin-inhibited respiration. These profiles are qualitatively comparable with those of the classical lipophilic weak-acid uncoupler, carbonylcyanide trifluoro-methoxyphenylene hydrazone (FCCP), which acts by promoting the electrogenic transport of H+ ions across mitochondrial membrane. PMID- 2992525 TI - Studies on the mechanism of action of omeprazole. AB - The effects of omeprazole on preparations of pig gastric (H+ + K+)-ATPase have been studied. Omeprazole was found to inhibit the (H+ + K+)-ATPase activity in a time-dependent manner. Inhibition was more pronounced at pH 6.1 compared with pH 7.4 and decreased as the concentration of (H+ + K+)-ATPase preparation increased. The potency of omeprazole was therefore highly dependent upon the conditions used. When pre- incubated with (H+ + K+)-ATPase preparation (30 micrograms protein/ml) for 30 min at 37 degrees and pH 6.1, omeprazole inhibited the (H+ + K+)-ATPase activity with an IC50 of 3.9 microM. This inhibition was shown to be irreversible in nature. Whilst omeprazole itself was not very potent as an inhibitor of the (H+ + K+)-ATPase activity at pH 7.4 (IC50 = 36 microM), transient acidification of omeprazole resulted in the formation of a compound(s) which produced marked inhibition at this pH (IC50 = 5.2 microM). The effects of omeprazole in the absence of acidification may have resulted from the rate limiting formation of this compound. Radiolabelled omeprazole was shown to incorporate into the (H+ + K+)-ATPase preparation in a time-dependent and pH dependent manner. Omeprazole, radiolabelled in three separate positions (the sulphur atom and the two adjacent carbon atoms), incorporated with equivalent time courses suggesting that the incorporation did not involve a fragmentation of the omeprazole molecule. Under conditions shown to produce a 50% inhibition of (H+ + K+)-ATPase activity, [14C] omeprazole had incorporated to a level of 4-5 nmoles/mg protein. Incorporation continued beyond the point required to produce 100% inhibition of (H+ + K+)-ATPase activity and reached 30 nmoles/mg protein after 5 hr. Prior acidification of the omeprazole resulted in a more rapid initial rate of incorporation although the final level of incorporation was lower than for omeprazole. Omeprazole was also shown to interact with the (Na+ + K+) ATPase from dog kidney. Omeprazole inhibited the (Na+ + K+)-ATPase activity (IC50 = 186 microM). Acid-degraded omeprazole inhibited the (Na+ + K+)-ATPase activity with greater potency (IC50 = 19 microM) and was also shown to incorporate into this enzyme preparation. PMID- 2992526 TI - Bis-(diethyldithiocarbamato) copper complex: a new metabolite of disulfiram? PMID- 2992527 TI - Selective inhibition of one of the cyclic AMP phosphodiesterases from rat brain by the neurotropic compound rolipram. PMID- 2992528 TI - Microsomal reduction of bisulfite (aqueous sulfur dioxide)--sulfur dioxide anion free radical formation by cytochrome P-450. PMID- 2992529 TI - Polyarthritis as the presenting symptom of a malignant fibrous histiocytoma of the heart. PMID- 2992530 TI - Suloctidil: long-term trial in the aged. Study of microcirculation, viscosity, red cell deformability, fibrinolysis and lipid fractions. AB - 15 aged patients with clinical and laboratory evidence of cerebrovascular insufficiency and/or myocardial sclerosis were given suloctidil (Locton) at the daily dose of 200 mg X 3, for 180 days, under the conditions of an open trial. Blood and plasma viscosity, red cell deformability, total lipids, cholesterol, triglycerides, fibrinolysis (plasminogen, antiplasmin, euglobulins with and without activator, fibrinogen), and capillaroscopic parameters in the small conjunctival vessels (artery, vein, and capillary diameter, appearance of collaterals, red cell aggregation and flow homogeneity) were evaluated. After 30 days of treatment and particularly at the end of the trial a significant improvement of almost all these parameters was observed. Tolerance was always excellent. PMID- 2992531 TI - [Hexoprenaline--a new tocolytic for treatment of premature labor]. AB - Hexoprenaline is a selective beta 2-mimetic drug used in tocology for the prevention of premature labor and immature birth. From clinical application of the drug in bronchospasm therapy its selectivity, which is due to the elongated nitrogen substituent, is well-known. Because of the relatively small stimulation of cardiac beta 1-receptors the side-effects related therewith are less pronounced than with other beta-mimetics. The extent of tachycardia depends on the initial sympathomimetic condition of the patients. The success rate for hexoprenaline tocolyses is 34-78%, dependent on the initial tocological condition. The advantage of hexoprenaline compared with other tocolytics on the basis of experience made so far seems to relate to the smaller increase in chronotropy and the better tolerability. PMID- 2992532 TI - [Improved immunocompetence in two children with chronic Epstein-Barr virus infection under treatment with a standardized thymus hormone preparation (thymostimulin)]. AB - Two children with chronic Epstein-Barr virus infection were treated with a standardized thymic hormone preparation (thymostimulin, TP-1 Serono). The treatment was well tolerated. No adverse reactions, no side effects were observed. Results of immunological investigations showed a trend towards normalisation. Additional clinical evidence showed up, which pointed to a response to the therapy. PMID- 2992533 TI - Effect of ryodipine on electromechanical parameters of heart and vessels, cAMP phosphodiesterase activity and swelling-contraction cycle of mitochondria. AB - 2,6-Dimethyl-3,5-dimethoxycarbonyl-4-(o-difluoromethoxyphenyl)-1,4 dihydropyridine (ryodipine, PP-1466), an effective Ca2+ channel blocker, diminishes contraction force and decreases duration of action potential in the frog heart ventricle strips. Dissociation constants K0.5 are 2 x 10(-7), 5 x 10( 7), and 10(-6) mol/l for PP-1466, nifedipine and nicardipine, respectively (at 0.25-0.3 Hz stimulation). One molecule of PP-1466 or nifedipine apparently interacts with two receptors on the channel (n = 0.5), nicardipine with one receptor (n = 1). The binding energy of PP-1466 and nifedipine increases at closed and diminishes at open channels which is in contrast to nicardipine, whose effect is irreversible. Thus, the site of nicardipine action differs from that of PP-1466 and nifedipine. PP-1466 (10(-8) mol/l--10(-6) mol/l) suppresses contraction force and diminishes frequency of spontaneous contractions of the rabbit atria, and also displays antagonism to the effect of Ca2+ upon rabbit auricle contractions. In the isolated rabbit aorta and portal vein PP-1466 is more antagonistic to contractions caused by Ca2+ than by epinephrine. Both competitive and non-competitive types of antagonism can be distinguished.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992534 TI - Influence on serum lipids, lipoproteins and blood pressure of mackerel and herring diet in patients with type IV and V hyperlipoproteinemia. AB - Eight patients with type IV and V hyperlipoproteinemia were put on a mackerel and herring diet of an isocaloric regimen for 2 weeks, in a cross-over design. At the end of the dietary periods a predominant increase of eicosapentaenoic acid (EPA - C20:5, n-3) in cholesterol esters and of docosahexaenoic acid (DHA - C22:6, n-3) in serum triglycerides, being more pronounced after mackerel as compared to herring diet, could be confirmed. After mackerel diet serum triglycerides and total cholesterol were significantly lower, returning to basal levels 3 months later. High density lipoprotein (HDL) cholesterol appeared slightly increased after mackerel diet and decreased to initial values thereafter. After herring diet, which contained half as much EPA as compared to mackerel diet, the differences were minor. The decline of free fatty acids (FFA) and insulin at the end of the mackerel period reached the level of significance 60 min and 120 min, respectively, after glucose load. A significantly lower systolic blood pressure in recumbent and upright position after the mackerel period could be found, whereas diastolic pressure and blood pressure after herring diet remained unchanged. PMID- 2992535 TI - Plasma lipid and lipoprotein cholesterol levels in swine. Modification of protein induced response by added cholesterol and soy fiber. AB - Piglets, aged 8 weeks and weighing 12-18 kg, were fed semi-purified casein or soy protein diets, with or without cholesterol and soy hull fiber, for 2 months. In addition to observing the effects of the dietary treatments on growth, the modification of the primary hypocholesterolemic action of soy protein by cholesterol and soy fiber was studied. Pigs fed the soy protein or casein diets grew normally with no difference in weight gain. Plasma triglyceride and phospholipid levels, as well as several plasma metabolic indices examined, were not significantly affected by dietary treatment. However, plasma total cholesterol was higher (but not significantly) in pigs fed casein than in those fed soy protein alone. Cholesterol feeding induced markedly significant (P less than 0.05) hypercholesterolemia with either protein source, compared to feeding the proteins without added cholesterol. Dietary soy fiber fed simultaneously with cholesterol decreased the cholesterol-induced hypercholesterolemia, but the reduction was significantly greater (P less than 0.05) with soy protein than with casein in the diet. Analyses of the lipoprotein cholesterol indicated that LDL cholesterol was much more sensitive to the changes induced by feeding cholesterol and soy fiber than either HDL or VLDL cholesterol. These findings suggest a beneficial role of dietary soy fiber in hypercholesterolemia. PMID- 2992536 TI - Input-output relationships of identified buccal neurones involved in feeding control in Aplysia. AB - A group of about 28 neurones located in the lateral portion of the caudal face of Aplysia buccal ganglion and projecting into the cerebro-buccal connective were identified by retrograde cobalt staining, and designated as L neurones. It was found that the L neurones did not establish synaptic relations with the known buccal neurones, which are mainly involved in the production of the consummatory phase of feeding, nor with several cerebral neurones tested, including the well known serotonin giant cell. Neither did they show responses to stimulation of the nerves directed to the buccal mass. On the other hand, the L neurones showed depolarizing responses, with the possible addition of a weak, slower hyperpolarizing phase, to stimulation of the ipsi- and contralateral oesophageal nerves, which innervate the portion of the gut posterior to the buccal mass. These findings, together with several properties of the oesophageal nerve input, suggest that one function of the L cells is to transmit information about gut regions posterior to the buccal mass towards the cerebral ganglia, and that they may mediate the inhibitory influence which in Aplysia is known to be exerted upon feeding by the presence of bulk in the anterior gut. The L neurones showed synaptic responses - consisting mainly or exclusively of depolarizations - to stimulation of the cerebro-buccal connectives. Besides this, large, tonic EPSPs, which often occurred in the 'spontaneous' activity of the L neurones, were found to be generated by spikes that travelled in the cerebro-buccal connective towards the buccal ganglion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992537 TI - Neocortical involvement in the acquisition and retention of learned alcohol aversions in rats. AB - In one experiment, rats lacking gustatory neocortex (GN) were compared with control (both normal and control lesion) rats in the acquisition of a learned alcohol aversion. In a second experiment, the effect of GN ablations on preoperatively learned alcohol aversions was examined. Results showed that rats lacking GN both learned and retained alcohol aversions in a normal manner. These same GN rats, however, extinguished the alcohol aversions significantly faster than controls. These results are in contrast to previous data which indicated that the integrity of the GN was necessary for normal acquisition and retention of learned taste aversions. It is suggested that the odor qualities present in the alcohol stimulus account for the relatively normal performance of GN rats. PMID- 2992539 TI - Steroid conversions by the 19-day old foetal rat ovary in organ culture. AB - The ovaries from 19-day old foetal rats were cultured in vitro on Wolff and Haffen's semi-solid medium with radioactive progesterone, dehydroepiandrosterone, testosterone or androstenedione added as precursors, and in the presence or absence of lutropin, follitropin and human chorion gonadotropin. After a 24-h culture period, the media were analysed for the presence of testosterone, androstenedione, oestrone and oestradiol, which were measured by specific activity determination after isotopic dilution. PMID- 2992540 TI - The use of silica gel in evidential tests in drunken drivers: problems related to alcohols adsorption and elution. PMID- 2992538 TI - Isolation of a cAMP receptor protein from yeast mitochondria (Mr 45000) and comparison with mitochondrial RNA polymerase (Mr 45000). AB - We have isolated a cAMP-binding protein from highly purified yeast mitochondria by affinity chromatography. It is a lipophilic protein of molecular mass 45 000 Da, which is tightly membrane-bound and localized on the outer surface of the inner membrane. It can be solubilized in active form under mild conditions. The cAMP receptor resembles mitochondrial RNA polymerase prepared as described by Levens et al. [(1981) J. Biol. Chem. 256, 1474] in a surprisingly large number of properties including molecular mass. Comparison of the two proteins revealed that the polypeptide previously considered as RNA polymerase is, in fact, a mitochondrial cAMP receptor protein. PMID- 2992541 TI - Relevance of anti-T monoclonal antibodies in the study of children with primary immunodeficiencies. AB - T-cell phenotypic analysis with anti-T monoclonal antibodies (MoAb) was performed on 37 children with immunologic disorders. Abnormalities of T-cell differentiation and/or of T-cell subset distribution were observed in many patients. In particular two infants with severe combined immunodeficiency showed immunologically incompetent common thymocytes (OKT6+) in the circulation, in one case a proportion of OKT6+ cells was OKT4-, OKT8-. A boy with a selective T-cell defect synthetized normal levels of Ig classes, despite the marked reduction of helper/inducer T-cells (OKT4+). Irregularities of T-cell subsets were also noted in children with Wiskott-Aldrich syndrome and in some patients with selective IgA defect or hypogammaglobulinaemia. In one of these, in whom the agammaglobulinaemia was caused by EBV infection, a persistently reversed OKT4+/OKT8+ ratio together with an excessive suppressor T-cell function were found more than 10 years after the onset of the disease. Such a case supports the hypothesis that a viral infection may cause, in a predisposed host, both the agammaglobulinaemia and an abnormality of the regulatory T-cell subpopulations. Such abnormalities, together with those found in the other children studied, underline the importance of MoAb against different T-cell antigens for a better characterization of primary immunodeficiencies. PMID- 2992542 TI - [Reduction of cytochrome c by NADH induced by light]. AB - We have studied the behaviour of Fe(III) cytochrome c upon irradiation in the 290 360 nm wavelength range either in the presence or in the absence of NADH; in both cases the photoexcitation caused the reduction of the heme iron. When the irradiation was performed in the absence of NADH, the iron reduction was coupled to a non reversible modification in the protein structure; the photoreduction quantum yield was decreasing with the increase of the irradiation wavelength. Irradiation in the presence of NADH gave heme iron reduction coupled to NADH oxidation and the protein resulted finally unmodified; the quantum yield depended on the irradiation wavelength in a way similar to the observed in the absence of NADH, but it was tenfold higher. We propose that in both cases the active species is an electronic excited state of the heme iron. PMID- 2992543 TI - Partial purification and some properties of a 5'-nucleotidase from chicken liver with activity toward pyrimidine nucleoside monophosphates. AB - A 5'-nucleotide phosphohydrolase activity, purified about 200-fold from cytosol of chicken liver, prefers as substrates pyrimidine nucleoside monophosphates, particularly the cytidilic forms. This activity, independent by divalent metal ions, shows a pH optimum of 6.2 and an isoelectric point of 5.2. PMID- 2992544 TI - [Modulating action of barium and other ions on the sensory activity of frog labyrinth]. AB - The effect of Ba2+, TEA, 4-AP and CoCl2 on the EPSP and spike discharges recorded from single fibres of the posterior nerve in the isolated frog labyrinth has been investigated. In Ca-free solution Ba2+ preserved, at low concentration (0.3 mM), the resting activity and at higher levels (up to 6 mM) it resulted in a pronounced facilitation of the EPSP and spike discharges. Facilitation increased on increasing Ba2+ concentration up to 4-5 mM and it was more evident in those units exhibiting a low resting spike firing. The effect of Ba2+ (1 mM) was completely antagonized by 10 mM Ca2+ X CoCl2 (3 mM) suppressed the resting rate at the normal external Ca2+ concentration; the Co2+ block was partially relieved by 1.8 mM Ba2+ X TEA (20 mM) evoked a clear-cut increase in the EPSP and spike discharges which, however, was less consistent than that produced by Ba2+. By comparing the effect of TEA on the spike frequency with that obtained at different Ba2+ levels, the Ba2+ capacity to carry the Ca2+ current was dissected. Such an effect is dose-dependent and it is more evident in low-frequency units. Conversely, 4-AP did not affect the resting discharge frequency. These results indicate that either the Ca2+ or the Ba2+ current sustain the transmitter release at the cyto-neural junction. The effect of TEA suggests that the Ca2+-dependent K+ current may play an important role in supporting the neurosecretory process by controlling the membrane potential of the hair cells. PMID- 2992545 TI - [Morphological, ultrastructural, cytochemical and functional analysis of neutrophils deficient in myeloperoxidase]. AB - Eighteen patients with established hereditary myeloperoxidase deficiency underwent morphological, ultrastructural, cytochemical and functional analysis in order to correlate the lack of peroxidase from phagocytes with other leucocyte activities. Cytochemical and ultrastructural findings only confirmed the peroxidase defect in neutrophil and monocyte population, whereas normal peroxidase activity was detected in eosinophil granulocytes ("Alius-Grignaschi anomaly"). Morphological analysis, as determined by both ligh and electron microscopy, showed in two patients with total MPO-deficiency a large number of neutrophils (50-60%) with nuclear abnormalities very similar to Pelger-Huet's heterozygous form (two lobed neutrophils having a typical pince-nez appearance and a nuclear chromatin coarser than that of normal PMNL). Other 2 cases displayed a 50-60% five-lobed neutrophils, as occur in congenital nuclear hypersegmentation of PMNL. These findings suggest that Alius-Grignaschi anomaly and Pelger-Huet syndrome can be found associated in the same individuals, since both these abnormalities have a genetic origin. Finally, since an impaired bactericidal and fungicidal activity was observed, no patients displayed particular susceptibility to persistent or severe infections, thus confirming the presence of MPO-independent enzymatic systems. PMID- 2992546 TI - [Effect of ACTH on water content of rat brain]. PMID- 2992547 TI - Naturally occurring anticarcinogenic substances in foodstuffs. PMID- 2992548 TI - Vitamin E: interactions with free radicals and ascorbate. PMID- 2992549 TI - Mechanisms and consequences of lipid peroxidation in biological systems. PMID- 2992551 TI - Simple versus complex carbohydrate use in the diabetic diet. PMID- 2992550 TI - Brown adipose tissue metabolism and thermogenesis. PMID- 2992552 TI - [Fenestration of the internal carotid artery in the neck]. AB - Fenestration or duplication of the internal carotid artery is extremely rare. A review of the literature revealed only two such cases by angiography. We present two further cases of internal carotid artery fenestration, incidentally disclosed by angiography. The embryological basis and the clinical significance of this anomalous condition were discussed. Case 1. A 47-year-old woman was admitted to hospital on May 13, 1982, after the first attack of subarachnoid hemorrhage. Four vessel angiography showed a saccular aneurysm arising at the trifurcation of the right middle cerebral artery. In addition, a fenestration of the right internal carotid artery was demonstrated at the level of the atlas: the cervical portion of the vessel distal to the level of C-2 was markedly elongated and dilated in caliber. Moreover, the right external carotid artery occupied a dorso-lateral position relative to the internal carotid artery. Case 2. A 51-year-old man was admitted to hospital on April 20, 1983, for evaluation of grand mal seizures. CT scans and four-vessel angiograms showed a malignant glioma of the right temporal lobe. Additionally, a fenestration of the right internal carotid artery was identified at the level of the atlantoaxial joint, and the vessel was dilated in caliber at the level of C-1 to C-2. PMID- 2992553 TI - [Patient with progressive multifocal leukoencephalopathy by in situ hybridization: comparison of in situ hybridization using isotopic and biotinylated probes]. AB - Ward et al developed biotinylated probes to hasten virus nucleic acid detection. We used these probes to trace SV 40 and JC virus nucleic acids in PML brain. This procedure visualizes the hybridization of viral sequences by affinity cytochemistry with avidin-biotin peroxidase complexes and diaminobenzidine. In the PML frozen white matter numerous oligodendroglial nuclei hybridized with JC virus biotinylated and tritium labelled probes in areas adjacent to active demyelination. Although tritium labeled probes was most sensitive and gave lowest background this biotinylated method showed be useful as a rapid diagnostic test for the specific detection of viral nucleic acid sequences in brain tissues from patients with central nervous system (CNS) infectious diseases. PMID- 2992554 TI - [NMR: basic and clinical researches in diagnosis of CNS lesions]. PMID- 2992555 TI - [MR imaging of intracranial tumors by superconducting system operating at 0.35 tesla]. PMID- 2992556 TI - [Postnatal development of barreloid neuropils in the ventrobasal complex of mouse thalamus: a histochemical study for cytochrome oxidase]. AB - Postnatal development of barreloid neuropils was studied in the ventrobasal complex of mouse thalamus. A single barreloid neuropil which topographically represented a corresponding facial vibrissa on the contralateral side was demonstrated as a solid reaction mass with the method for cytochrome oxidase. This architectural unit was called a vibrissa compartment. On postnatal Day 0, the nuclear area of the ventrobasal complex showed higher activity for the enzyme than in regions around, but no barreloids were seen. Barreloid neuropils became detectable on postnatal Day 3 in the dorsomedial subnucleus of the ventrobasal complex and the septal delimitation of barreloids was established on postnatal Day 6. The arrangement of barreloids showed a distinct orderlines similar to that of facial vibrissae on a horizontal section. Barreloid neuropils contained synaptic glomeruli ensheathed with glial processes, in which dendritic processes and their excrescences of nerve cells received large terminal knobs in well organized manner. On postnatal Day 10, immature glomerular forms were found in barreloid neuropils. The synaptic glomerulus of mature type, however, was not formed till postnatal Day 13. The number of synaptic junctions was examined in barreloid neuropils from postnatal Day 3 to 21. The result that the growth rate of synaptic junctions was the highest in a short period from postnatal Day 10 to 13 suggested rapid maturation of barreloid neuropils in this period. On postnatal Day 21, several types of synaptic contacts became remarkable in matured glomeruli. One of them, a filamentous type with association of clear synaptic vesicles at presynaptic sites was indicated. And non-synaptic junctional complex accompaning tubular structures beneath dense materials at dendritic sites was remarked between terminal knobs and dendritic processes. From results the following conclusions are remarked that at the level of mouse thalamus, septal delimitation of vibrissa compartments appeared on postnatal Day 3 and was established until postnatal Day 6. On the other hand, barreloid neuropils containing glomerular synaptic complexes became a maturing state on postnatal Day 13, that is, about one week later after architectural compartmentalization. PMID- 2992557 TI - The effects of an airbrasive instrument on dental hard tissues, skin and oral mucosa. PMID- 2992558 TI - Differential effect of platelet inhibitors in normal and in hypercholesterolaemic subjects. AB - Dibutyryl cyclic AMP, forskolin, dipyridamole and butyl imidazole inhibited platelet aggregation (induced by ADP or collagen) in washed platelets more than in platelet-rich plasma preparations. Aspirin, indomethacin and epoprostenol (prostacyclin, PGI2) showed no preferential inhibition of these platelet preparations. When platelet-rich plasma from either normal or familial hypercholesterolaemic (FH) subjects was used, aspirin, indomethacin and dipyridamole (but not forskolin) inhibited platelet aggregation in normal subjects more than in FH patients. When low doses of aspirin (75 mg daily for 7 days) or dipyridamole (250 mg, single dose) were administered in vivo, platelet aggregation was inhibited more in the normal subjects in comparison to the patient group. PMID- 2992559 TI - Involvement of calcium in the caffeine stimulation of human sperm motility. AB - The shape of the concentration-response curve and the amplitude of maximal motility increase for the stimulatory effect of caffeine on human sperm motility were similar to those of EGTA, a calcium chelator, and lanthanum chloride, a calcium entry blocker. On the other hand, the maximal motility stimulation induced by dibutyryl cyclic AMP was similar to that induced by trifluoperazine, a calmodulin antagonist. The antagonism of A23187 on the motility-stimulating effect of EGTA and caffeine was competitive in nature, while that on cyclic AMP was non-competitive. We proposed that there could be two mechanisms for caffeine to stimulate sperm motility: at higher concentrations, caffeine modified calcium translocation; at lower concentrations, caffeine increased cyclic AMP level. The importance of calcium in the regulation of human sperm motility was reconfirmed in this study. PMID- 2992560 TI - Plasma melatonin during desmethylimipramine treatment: evidence for changes in noradrenergic transmission. AB - Plasma melatonin was used to determine overall beta-adrenergic transmission through pineal neuro-effector junctions during desmethylimipramine (DMI) treatment in 10 normal subjects. Changes in plasma melatonin indicated that an initial increase in noradrenaline (NA) transmission produced by DMI was counteracted by adaptive changes which restored transmission to normal by the third week of treatment. A 'rebound' increase in NA transmission was seen on DMI withdrawal. The results suggest that the adaptive changes which occur in NA synapses during DMI treatment do not, as has been proposed, decrease NA transmission below normal levels, but instead restore homeostasis in the presence of the drug. PMID- 2992561 TI - The effect of captopril on the reflex control heart rate: possible mechanisms. AB - Angiotensin converting enzyme inhibitors reduce blood pressure without reflex tachycardia, possibly as a result of enhanced hypothesis that this results from the removal of the parasympathetic activity. We examined the vagolytic action of angiotensin II or alternatively by acetylcholinesterase inhibition. Both captopril and [Sar1ala8] angiotensin II, (saralasin), caused modest falls in blood pressure, without increasing heart rate in normotensive subjects. Captopril and saralasin significantly attenuated the vagally mediated heart rate slowing after facial immersion in water. There was a close correlation between the effects produced by captopril and saralasin on the diving reflex. Infusion of subpressor doses of angiotensin II, reversed the hypotensive effect of captopril and returned the bradycardia after facial immersion to placebo level. In vitro neither captopril nor enalapril or lisinopril affected bovine erythrocyte acetylcholinesterase activity. The parasympathetic effect of angiotensin converting enzyme inhibitors appear to reflect a direct consequence of the removal of angiotensin II. PMID- 2992562 TI - A dose-response study of HOE 498, a new non-sulphydryl converting enzyme inhibitor, on blood pressure, pulse rate and the renin-angiotensin-aldosterone system in normal man. AB - The effect of different oral doses of HOE 498, a new non-sulphydryl containing converting enzyme inhibitor, was investigated in a double-blind, placebo controlled study in normotensive volunteers. Dose-related reductions in serum converting enzyme activity, plasma angiotensin II and aldosterone were seen, greater at 4 h than at 12 h after drug ingestion. Converse dose-related increases in blood angiotensin I and plasma active renin concentration occurred. Falls of angiotensin II were as great with 20 mg as with 50 mg of HOE 498, although the effect was more prolonged with 50 mg. The reductions in concentrations of plasma angiotensin II and serum converting enzyme activity and the increases in plasma renin concentration were correlated with the concentration of HOE 498 - diacid in plasma. Dose-related falls in both supine and erect blood pressure were maximal 2 3.5 h after dosing. Pulse rate increased marginally but insignificantly in the supine; slightly and significantly in the upright position, concomitantly with the blood pressure reduction at all doses of active drug. We conclude that effects of single doses of HOE 498 on the renin-angiotensin system are maximal within 4 h, but are still apparent after 24 h. Thus it is likely that once daily administration will be adequate for treatment of high blood pressure in patients. PMID- 2992563 TI - Inhibition of methoxamine-induced bronchoconstriction by ipratropium bromide and disodium cromoglycate in asthmatic subjects. AB - We compared the effects of pretreatment with saline, ipratropium bromide, and disodium cromoglycate (DSCG) on bronchoconstriction induced by methoxamine--an alpha-adrenoceptor agonist, in asthmatic subjects. All 12 patients bronchoconstricted in response to methoxamine after saline. The PD20 (the dose of methoxamine causing a 20% fall in forced expiratory volume in 1 s [FEV1]) ranged from 0.3-18 mumol. Ipratropium bromide (200 micrograms administered by aerosol) significantly inhibited (P less than 0.05) the response to methoxamine in all patients without producing significant changes in the mean baseline lung function. The mean PD20 for methoxamine after saline was 6.8 mumol and 95% confidence limits (CL) were 3.6, 12.7 mumol. The mean PD20 for methoxamine after ipratropium bromide was 35.4 (95% CL 28.8, 43.6) mumol. DSCG also produced significant (P less than 0.05) shifts to the right in the methoxamine dose response curves, but did not affect resting airway calibre as measured by the FEV1. The mean PD20 for methoxamine increased from 3.3 mumol (95% CL 1.1, 10.0 mumol) after saline to 25.1 mumol (95% CL 14.1, 44.6) after DSCG pretreatment. These findings suggest that alpha-adrenoceptors in the airways of asthmatic subjects may be located at sites other than smooth muscle--possibly on mast cells but more likely on nerve endings and/or parasympathetic ganglia. PMID- 2992565 TI - The neurological basis of anxiety. PMID- 2992564 TI - Effects of nicardipine on aldosterone release and pressor mechanisms. AB - This study evaluated the effects of nicardipine, following intravenous infusion and oral administration, on the pressor and aldosterone responses to infused angiotensin II. Six healthy, normotensive male subjects were studied. Following administration of nicardipine, no significant change in blood pressure was seen. Nicardipine attenuated the pressor response produced by intravenous administration of angiotensin. Nicardipine did not inhibit the aldosterone response to angiotensin. PMID- 2992566 TI - Role for different cell proteinases in cancer invasion and cytolysis. AB - The crucial role of non-plasminogen dependent serine proteinases is tissue invasive and cytolytic functions of Walker 256 cancer cells has been documented using a rat urinary bladder invasion and a 125I-labelled fibroblast cytolysis assay. The invasive capacity of these cancer cells was abrogated by non toxic concentrations of the serine proteinase inhibitors, diisopropylfluorophosphate and phenylmethylsulfonylfluoride, but not by metallo or cysteine proteinase inhibitors. Although tumour cell collagenase activity and plasminogen activator were demonstrated, these proteolytic enzymes were not essential in these in vitro assays. These results suggest that different categories of proteinases play specific roles in the complicated process of cancer invasion. PMID- 2992567 TI - Transplantability in nude mice of embryonic and other childhood tumours. PMID- 2992568 TI - The induction of pulmonary phospholipidosis and the inhibition of lysosomal phospholipases by amiodarone. AB - Administration of high doses of amiodarone to young adult rats leads to phospholipidosis of the lung, with extensive phospholipid storage by type II pneumonocytes and alveolar macrophages. Biochemical analysis reveals an increase in the total phospholipid content of the lung and in the proportion of phosphatidylcholine. The cause of the phospholipidosis is suggested to be the inhibition of lysosomal phospholipases, responsible for catabolizing phospholipids. It is shown that amiodarone is a potent inhibitor of phospholipases prepared from the soluble fraction of adult rabbit lung lysosomes. PMID- 2992569 TI - The chemiluminescent response of human phagocytic cells to mineral dusts. AB - Luminol-dependent chemiluminescence (CL) was used to assess the in vitro production of reactive oxygen species by human neutrophils and monocytes on exposure to six standard respirable mineral dusts. Every dust caused CL production in both phagocytic cell types, although, for each dust, the two cells showed a different pattern of response. Light output was markedly affected by the presence of serum in the system. While the results illustrated the complexity of the interaction between mineral dusts and monocytes and neutrophils, they did not support the hypothesis that pathogenic dusts would induce the production of more reactive oxygen species than non-pathogenic dusts. PMID- 2992571 TI - Exchange rates and numbers of annular lipids for the calcium and magnesium ion dependent adenosinetriphosphatase. AB - A spin-labeled phospholipid is used to study lipid-protein interactions in the (Ca2+,Mg2+)-ATPase of sarcoplasmic reticulum from muscle. A novel null method is used to decompose composite electron spin resonance spectra into two components, characteristic of immobilized and mobile environments. Calculations based on a random mixing model suggest that protein-protein interactions will be relatively rare in these systems and that the immobilized lipid does not represent lipid trapped between proteins but rather represents annular phospholipid at the lipid protein interface of the adenosinetriphosphatase. The apparent decrease in the amount of immobilized lipid with increasing temperature is shown to be consistent with lipid exchange between bulk and annulus, characterized by an exchange time of 10(-7) s at 37 degrees C. A minimum number of annular phospholipid sites of 32 and 22 are calculated at 0 and 37 degrees C, respectively. PMID- 2992570 TI - Effects of experimental joint inflammation on bone marrow and periarticular bone. A study of two types of arthritis, using variable degrees of inflammation. AB - The effects of joint inflammation on bone marrow and periarticular bone were studied in mice using antigen-induced arthritis and zymosan-induced arthritis as models for an immune and a non-immune-mediated chronic inflammation. To allow for a comparison of the two types of arthritis care was taken to induce comparable degrees of inflammation as evaluated with 99mTechnetium uptake and histology. The antigen-induced arthritis caused a significant suppression of the mitotic activity in the bone marrow close to the inflammatory focus during the first days of arthritis. The zymosan-induced arthritis did not produce alterations of the bone marrow activity. Both types of arthritis were able to induce long-lasting and irreversible damage to cartilaginous and bony structures. Apposition of bone was observed in both types of arthritis although much earlier in the antigen induced arthritis. The apposition of bone was found to emerge largely from the periosteum and not from the epiphyseal plates, as shown by 125I-deoxyuridine autoradiography. Qualitative and quantitative differences suggest that joint inflammation which is immunologically mediated, results in more severe (peri)articular tissue damage than a non-immunological arthritis. PMID- 2992572 TI - Rate of transmembrane electron transfer in chromaffin-vesicle ghosts. AB - The chromaffin vesicle of the adrenal medulla contains a transmembrane electron carrier that may provide reducing equivalents for dopamine beta-hydroxylase in vivo. This electron-transfer system can be assayed by trapping ascorbic acid inside resealed membrane vesicles (ghosts), adding an external electron acceptor such as ferricytochrome c or ferricyanide, and following the reduction of these acceptors spectrophotometrically. Cytochrome c reduction is more rapid at high pH and is proportional to the amount of chromaffin-vesicle ghosts, at least at low ghost concentrations. At pH 7.0, ghosts loaded with 100 mM ascorbic acid reduce 60 microM cytochrome c at a rate of 0.035 +/- 0.010 mu equiv min-1 (mg of protein)-1 and 200 microM ferricyanide at a rate of 2.3 +/- 0.3 mu equiv min-1 (mg of protein)-1. The rate of cytochrome c reduction is accelerated to 0.105 +/- 0.021 mu equiv min-1 (mg of protein)-1 when cytochrome c is pretreated with equimolar ferrocyanide. Pretreatment of cytochrome c with ferricyanide also causes a rapid rate of reduction, but only after an initial delay. The ferrocyanide-stimulated rate of cytochrome c reduction is further accelerated by the protonophore carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP), probably because FCCP dissipates the membrane potential generated by electron transfer. These rates of electron transfer are sufficient to account for electron transfer to dopamine beta-hydroxylase in vivo and are consistent with the mediation of electron transfer by cytochrome b-561. PMID- 2992574 TI - Conformation of spin-labeled tropomyosin in reconstituted muscle thin filaments in response to calcium ion and heavy meromyosin. AB - Tropomyosin (TM) exists in thermal equilibrium between a highly structured N state, a partially unfolded X state, and a completely unfolded D state, i.e., N in equilibrium X in equilibrium D. The strongly immobilized electron spin resonance (ESR) spectral component of spin-labeled TM corresponds to TM in the N state and the weakly immobilized component to TM in the X state below the main unfolding transition and to TM in the D state above this transition [Graceffa, P., & Lehrer, S. S. (1984) Biochemistry 23, 2606-2612]. The addition of actin, troponin (TN), and heavy meromyosin (HMM) to spin-labeled TM reduces the ratio of weakly to strongly immobilized labels, indicating a shift in the N in equilibrium X in equilibrium D equilibrium toward the N state. At 37 degrees C, for spin labeled TM alone K (=X/N) greater than 1.0 with some TM in the D state, K = 0.8 for spin-labeled TM bound to actin, and K less than 0.05 for spin-labeled TM bound to actin + TN +/- Ca2+, actin + HMM + TN +/- Ca2+, and actin + HMM. Thus, actin + TN dramatically shifts the TM structure to the N conformation with little further effect upon addition of Ca2+ or HMM. The temperature at which spin labeled TM begins to dissociate from a protein complex was determined from the temperature dependence of the ESR spectra.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992573 TI - Human cellular fibronectin: comparison of the carboxyl-terminal portion with rat identifies primary structural domains separated by hypervariable regions. AB - We report the isolation and characterization of four overlapping cDNA clones coding for human cellular fibronectin which continuously cover more than 3 kilobases in length. The nucleotide sequence of these cDNAs has been determined, thus elucidating the amino acid sequence of the C-terminal 794 residues of human fibronectin, which cover the edge of cellular-, heparin-, and fibrin-binding domains of this protein. Comparisons of the nucleotide sequences and the deduced amino acid sequences with those of rat [Schwarzbauer, J. E., Tamkun, J. W., Lemischka, I. R., & Hynes, R. O. (1983) Cell (Cambridge, Mass.) 35, 421] indicate a high degree of conservation at both nucleotide and amino acid levels. Comparison with previously published data on amino acid sequences of bovine fibronectin made it possible to identify structurally important features of the protein during the evolution of human, calf, and rat. The deduced human amino acid sequences contain five type III and three type I repeats of internal homologies. The interspecies conservation in amino acids is more pronounced in regions containing the internal repeats and within each functional domain. The implications of these interspecies conservation and divergence are discussed. PMID- 2992575 TI - Use of virus-attached antibodies or insulin molecules to mediate fusion between Sendai virus envelopes and neuraminidase-treated cells. AB - Anti-human erythrocyte antibodies or insulin molecules were covalently coupled to the glycoproteins (the hemagglutinin/neuraminidase and the fusion polypeptides) of Sendai virus envelopes with N-succinimidyl 3-(2-pyridyldithio)propionate and succinimidyl 4-(p-maleimidophenyl)butyrate as cross-linking reagents. Reconstituted Sendai virus envelopes, bearing covalently attached anti-human erythrocyte antibodies or insulin molecules, were able to bind to but not fuse with virus receptor depleted human erythrocytes (neuraminidase-treated human erythrocytes). Only coreconstitution of Sendai virus glycoproteins, bearing attached anti-human erythrocyte antibodies or insulin molecules with intact, untreated viral glycoproteins, led to the formation of fusogenic, targeted reconstituted Sendai virus envelopes. Binding and fusion of reconstituted Sendai virus envelopes, bearing anti-human erythrocyte antibodies or insulin molecules, with neuraminidase-treated human erythrocytes were blocked by the monovalent fraction, obtained after papain digestion of immunoglobulins, made of anti-human erythrocyte antibodies or free insulin molecules, respectively. The results of this work demonstrate an active role of the viral binding protein (hemagglutinin/neuraminidase polypeptide) in the virus membrane fusion process and show a novel and efficient method for the construction of targeted, fusogenic Sendai virus envelopes. PMID- 2992577 TI - Role of peptide structure in lipid-peptide interactions: high-sensitivity differential scanning calorimetry and electron spin resonance studies of the structural properties of dimyristoylphosphatidylcholine membranes interacting with pentagastrin-related pentapeptides. AB - The effects of amino acid substitutions in the pentapeptide pentagastrin on the nature of its interactions with dimyristoylphosphatidylcholine (DMPC) are assessed by differential scanning calorimetry and electron spin resonance. In two peptide analogues, the Asp at position 4 in pentagastrin (N-t-Boc-beta-Ala-Trp Met-Asp-Phe-NH2) is replaced by Gly or Phe. These uncharged, more hydrophobic peptides have little effect on the transition temperature of DMPC, but they broaden the transition and lower the transition enthalpy as do integral membrane proteins. These peptides also mimic the behavior of integral membrane proteins in decreasing the order of a 5-doxylstearic acid spin probe below the transition temperature and in exhibiting a second immobilized lipid component using a 16 doxylstearic acid spin probe in DMPC. Three charged peptides were studied: pentagastrin, an analogue with positions 4 and 5 reversed (i.e., ending in Phe Asp-NH2), and one with Asp replaced by Arg at position 4. All three of these charged peptides altered the phase transition behavior of DMPC to give two components, one above and one below the transition temperature of the pure lipid. With increasing peptide concentration, the higher melting transition became more prominent. The arginine-containing peptide produced the largest shifts in melting temperature followed by pentagastrin and then the "reversed" peptide. The arginine-containing peptide also increased the enthalpy of the transition. These peptides also increased the ordering of DMPC below the phase transition as measured with both 5- and 16-doxylstearic acid. The ordering effect was most pronounced with the arginine-containing peptide using the 5-doxylstearic acid probe. The results demonstrate that even the zwitterionic DMPC can interact more strongly with positively charged peptides than with negatively charged ones. In addition, peptide sequence as well as composition is important in determining the nature of peptide-lipid interactions. The markedly different effects of these pentagastrin peptides on the phase transition and motional properties of DMPC occur despite the similar depth of burial of these peptides with DMPC. PMID- 2992576 TI - Structure of the 3' portion of the bovine elastin gene. AB - A bovine genomic library constructed by partial Sau3A digestion and contained in lambda Charon 30 was screened by in situ hybridization with a 1.3-kilobase (kb) sheep elastin cDNA clone [Yoon, K., May, M., Goldstein, N., Indik, Z., Oliver, L., Boyd, C., & Rosenbloom, J. (1984) Biochem. Biophys. Res. Commun. 118, 261 269]. Three clones encompassing 10 kb of the bovine elastin gene were identified and characterized by restriction mapping and DNA sequencing of the 6.2 kb of the most 3' region of the gene. These analyses have permitted localization of eight exons in the 6.2 kb in which the translated exons vary in size from 27 to 69 base pairs, and there is an approximately 1-kb untranslated region at the 3' end. In addition to identification of sequences homologous to those found in porcine tropoelastin, the analyses defined a 58 amino acid sequence that forms the carboxy-terminal region of tropoelastin, and this sequence, which contains two cysteine residues, was previously not observed in the protein sequence data. The analyses also suggest that functionally distinct cross-link and hydrophobic domains of the protein are encoded in separate exons. PMID- 2992578 TI - Clostridium histolyticum collagenase: development of new thio ester, fluorogenic, and depsipeptide substrates and new inhibitors. AB - A new series of thio ester, depsipeptide, and peptide substrates have been synthesized for the bacterial enzyme Clostridium histolyticum collagenase. The hydrolysis of the depsipeptide substrate was followed on a pH stat, and thio ester hydrolysis was measured by inclusion of the chromogenic thiol reagent 4,4' dithiopyridine in the assay mixture. The best thio ester substrate, Boc-Abz-Gly Pro-Leu-SCH2CO-Pro-Nba, had a kcat/KM of 63 000 M-1 s-1, while several shorter thio ester sequences were inactive as substrates. In general, the peptide analogues of all the reactive thio ester substrates were shown to be hydrolyzed 5 10 times faster by collagenase. In one case (Z-Gly-Pro-Leu-Gly-Pro-NH2) where a comparison was made, the peptide substrate was respectively 8- and 106-fold more readily hydrolyzed than the corresponding thio ester and ester substrates. Cleavages of the two fluorescence-quench substrates Abz-Gly-Pro-Leu-Gly-Pro-Nba and Abz-Gly-Pro-Leu-SCH2CO-Pro-Nba could be easily followed fluorogenically since a 5-10-fold increase in fluorescence occurred upon hydrolysis. The fluorescent peptide substrate is the best synthetic substrate known for C. histolyticum collagenase with a kcat/KM value of 490 000 M-1 s-1. A series of new reversible inhibitors were developed by the attachment of zinc ligating groups (hydroxamic acid, carboxymethyl, and thiol) to various peptide sequences specific for C. histolyticum collagenase. The shorter peptides designed to bind to either the P3 P1 or P1'-P3' subsites were poor to moderate inhibitors. The thiol HSCH2CH2CO-Pro Nba had the lowest K1 (0.02 mM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992579 TI - Intramolecular electron and proton transfer in proteins: CO2- reduction of riboflavin binding protein and ribonuclease A. AB - The formate radical (CO2-) reacts with ribonuclease A to form the cystine disulfide radical as one of the products. CO2- reacts with the riboflavin binding protein of chicken egg white with the ultimate product being the neutral flavin semiquinone. Formation of the disulfide radical in ribonuclease is slower than the reaction between protein and CO2-; formation of the flavin semiquinone in the riboflavin binding protein is slower than the protein-CO2- reaction. We conclude for both proteins that CO2- must reduce an as yet unidentified group or groups, which in turn reduce(s) the disulfide of RNase or the flavin of riboflavin binding protein. This conclusion is supported in the case of ribonuclease by the observation of a transient, broad absorption band centered between 350 and 370 nm. The CO2--initiated reductions of the disulfide in ribonuclease and the flavin in the riboflavin binding protein are mixed first- and second-order processes. We propose that the transfer of an electron from the unknown intermediate(s) to the final product involves both inter- and intramolecular paths between groups that may not be in van der Waals contact. With the hydrated electron, in contrast to CO2-, as reductant of the riboflavin binding protein, the anionic semiquinone is observed as an intermediate. The anionic semiquinone is then rapidly protonated, yielding the stable neutral semiquinone. From the reaction kinetics and protein concentration dependence, we conclude that a group or groups on the protein donate(s) a proton to the anionic semiquinone by both inter- and intramolecular paths. PMID- 2992580 TI - Photoconversion of 7-dehydrocholesterol to vitamin D3 in synthetic phospholipid bilayers. AB - The incorporation of 7-dehydrocholesterol into synthetic phospholipid bilayers altered the distribution of products after photolysis. In liposomes, the relative amounts of 7-dehydrocholesterol and lumisterol were elevated, and tachysterol was reduced from the levels observed in hexane solution. Z to E isomerization of the previtamin to tachysterol is favored in organic solvents. The inhibition of this process is evidence that an ordered lipid matrix places a new constraint on the conformation of the ring B fission product--one in which the configuration is favorable for a return to a cyclized diene. Further, rate enhancements of up to 15-fold were observed for the thermal isomerization of the previtamin to vitamin D3 in liposomes. The free energies of activation for the reaction at 25 degrees C were reduced by 1.3-1.5 kcal/mol in the bilayer environment compared to that of hexane. As this reaction involves the concerted transfer of a hydrogen via a cyclic intermediate, it provides additional evidence for membrane stabilization of an all-cis conformation of the previtamin. Photoproduct ratios were also studied for 7-dehydrocholesterol adsorbed to a variety of solid supports. That nonspecific interactions of 7-dehydrocholesterol with lipid can influence product formation may have important implications with respect to the mechanism of vitamin D3 biosynthesis. PMID- 2992581 TI - Salt-dependent structural changes of neurohormones: lithium ions induce conformational rearrangements of ocytocin to a vasopressin-like structure. AB - The preferred average conformation and structural subdomain interactions of the nonapeptide hormones vasopressin and ocytocin have been analyzed through the determination of their hydrodynamic volume and the thermal coefficient of the frictional resistance to rotation of their tyrosine residue. A spherical gross shape and an ellipsoidal gross shape were assessed respectively for ocytocin and vasopressin by fluorescence polarization analysis. Investigation of the thermal coefficient of viscosity and the critical temperature of both hormones and analogues indicated that strong interactions hold together the two structural subdomains of ocytocin (the flexible six-membered ring and the COOH-terminal tripeptide tail). An opposite situation was found in the case of vasopressin where such interactions could not be detected between the rigid ring and the flexible COOH-terminal tail. Lithium ions were shown to promote ocytocin binding to specific neurophysin sites restricted, under standard conditions, to vasopressin. In the presence of lithium, the gross conformational shape of ocytocin becomes similar to that of vasopressin but in the absence of salt. In addition, the ocytocin ring becomes more rigid in the presence of lithium while decreasing interactions between the ring and the COOH-terminal tail were detected. It is proposed that lithium ions induce specific conformational rearrangements of ocytocin toward a vasopressin-like structure, allowing recognition of this hormonal ligand by a specific vasopressin binding domain of neurophysins. PMID- 2992582 TI - Proteolysis-associated deglycosylation of beta 1-adrenergic receptor in turkey erythrocytes and membranes. AB - A protease that can be inhibited by glutathione, dithiothreitol, and o phenanthroline but not by ethylenediaminetetraacetic acid converts the 50 kilodalton beta-adrenergic receptor in turkey erythrocyte membranes to a 40-kDa polypeptide which retains the specific ligand binding site. This conversion is attenuated in intact erythrocytes. The large 50-kDa peptide contains N-linked, complex carbohydrates and is retained on wheat germ agglutinin-Sepharose. The 40 kDa product of proteolysis does not bind to the wheat germ agglutinin and can thus be separated from the 50-kDa polypeptide by lectin chromatography. However, the large difference in molecular weights of the two receptor peptides cannot be accounted for solely by the different extent of glycosylation. PMID- 2992583 TI - The oxidation of cytochrome c oxidase by hydrogen peroxide. AB - The reaction of H2O2 with mixed-valence and fully reduced cytochrome c oxidase was investigated by photolysis of fully reduced and mixed-valence carboxy cytochrome c oxidase in the presence of H2O2 under anaerobic conditions. The results showed that H2O2 reacted rapidly (k = (2.5-3.1) X 10(4) M-1 X s-1) with both enzyme species. With the mixed-valence enzyme, the fully oxidised enzyme was reformed. On the time-scale of our experiments, no spectroscopically detectable intermediate was observed. This demonstrates that mixed-valence cytochrome c oxidase is able to use H2O2 as a two-electron acceptor, suggesting that cytochrome c oxidase may under suitable conditions act as a peroxidase. Upon reaction of H2O2 with the fully reduced enzyme, cytochrome a was oxidised before cytochrome a3. From this observation it was possible to estimate that the rate of electron transfer from cytochrome a to a3 is about 0.5-5 s-1. PMID- 2992585 TI - Na+-H+ exchange in cardiac sarcolemmal vesicles. AB - The transport of Na+ by a purified sarcolemmal vesicular preparation from canine ventricular tissue was studied as a function of both internal and external pH. The uptake of Na+ into sarcolemmal vesicles increased upon raising the extravesicular pH of the reaction medium. Half-maximal uptake of Na+ was observed at a pHo of about 8.1 and maximal uptake occurred at pH 8.6. The uptake of Na+ by sarcolemma was also dependent upon the intravesicular pH. Na+ uptake into sarcolemmal vesicles was greatly attenuated in the absence of a H+ gradient across the membrane. Transport of Na+ was potently inhibited by amiloride, a known blocker of Na+-H+ exchange. LiCl was also an effective inhibitor of Na+ transport. In the presence of optimal H+ gradients, Na+ uptake was linear for the first 5 seconds of the reaction and exhibited a Vmax of 290 nmol Na+/mg per min and a KNa of 3.5 mM. These experiments strongly indicate the presence of a Na+-H+ exchange system in cardiac sarcolemma. This activity appeared to be relatively specific for this membrane fraction. The identification of Na+-H+ exchange activity in a sarcolemmal vesicular fraction from the heart will permit extensive characterization of the regulation and kinetics of this antiporter in future investigations. PMID- 2992584 TI - Carboxyl group involvement in the meta I and meta II stages in rhodopsin bleaching. A Fourier transform infrared spectroscopic study. AB - Structural changes due to photoreceptor membrane bleaching can be studied by Fourier transform infrared difference spectroscopy [1,2]. In this paper we focus on the differences between rhodopsin and metarhodopsin I or II. Peaks in the 1700 1770 cm-1 region are observed, which may be produced by carbonyl groups in either carboxyl (COOH) or ester carbonyl (COOC) groups, the latter being found exclusively in membrane lipids. In order to distinguish between these two types of carbonyl groups, we have studied reconstituted membranes of rhodopsin in a synthetic phosphatidylcholine that lacks ester carbonyl groups. On this basis, we conclude that the major changes in this region are due to rhodopsin carboxyls which undergo either a change in local environment or a protonation/deprotonation reaction. Additional small changes in this region may reflect a direct involvement of phospholipids in the metarhodopsin I-to-II transition. One or more groups responsible for peaks near 1727 and 1702 cm-1 are inaccessible to the outside medium according to hydrogen/deuterium exchange. In contrast, carboxyl group(s) producing peaks near 1710, 1745 and 1768 cm-1 exchange freely with the outside medium and are therefore likely to be located near the membrane surface. Removal of a portion of the C-terminal tail region using proteinase K demonstrates that the carboxyl groups in the C-terminal sequence 248-348 are not involved directly in the rhodopsin to metarhodopsin II transition. At the meta I stage, only carboxyl peaks associated with buried groups appear, suggesting that the initial bleaching events, leading to the formation of this intermediate, produce structural rearrangements in the interior region of rhodopsin. These changes then spread to the peripheral surface regions during the metarhodopsin I to-II transition. PMID- 2992586 TI - Myometrial (Na+ + K+)-activated ATPase and its Ca2+ sensitivity. AB - Ouabain-sensitive (Na+ + K+)-ATPase activity in the rat myometrial microsome fraction could only be determined following detergent treatment. The (Na+ + K+) ATPase activity manifested by detergent treatment proved very stable even to high concentrations of NaN3, in contrast Mg+-ATPase activity was reduced to about 30 percent of the control. The major part of the Mg2+-ATPase in the myometrial membrane preparation was found to be identical with the NaN3-sensitive ATP diphosphohydrolase capable of ATP and ADP hydrolysis. This monovalent-cation insensitive ATP hydrolysis could be extensively reduced by DMSO. Furthermore DMSO prevented the inactivation of the (Na+ + K+)-ATPase activity. 10-100 microM Ca2+ inhibited the (Na+ + K+)-ATPase activity obtained in the presence of SDS by 15-50 percent. The Ca2+ sensitivity of the enzyme was considerably decreased if the proteins solubilized by the detergent had been separated from the membrane fragments by ultracentrifugation. The inhibitory effect could be regained by combining the supernatant with the pellet. Ca2+ sensitivity of the (Na+ + K+) ATPase activity was preserved even after removal of the solubilized proteins provided that DMSO had been applied. It appears that a factor in the plasma membrane solubilized by SDS may be responsible for the loss of Ca2+ sensitivity of the (Na+ + K+)-ATPase activity, the solubilization of which can be prevented by DMSO. PMID- 2992587 TI - Binding of DTNB to band 3 in the human red cell membrane. AB - Inhibition of red cell water transport by the sulfhydryl reagent 5,5'-dithiobis(2 nitrobenzoic acid) (DTNB) has been reported by Naccache and Sha'afi ((1974) J. Cell Physiol. 84, 449-456) but other investigators have not been able to confirm this observation. Brown et al. ((1975) Nature 254, 523-525) have shown that, under appropriate conditions, DTNB binds only to band 3 in the red cell membrane. We have made a detailed investigation of DTNB binding to red cell membranes that had been treated with the sulfhydryl reagent N-ethylmaleimide (NEM), and our results confirm the observation of Brown et al. Since this covalent binding site does not react with either N-ethylmaleimide or the sulfhydryl reagent pCMBS (p chloromercuribenzenesulfonate), its presence has not previously been reported. This covalent site does not inhibit water transport nor does it affect any transport process we have studied. There is an additional low-affinity (non covalent) DTNB site that Reithmeier ((1983) Biochim. Biophys. Acta 732, 122-125) has shown to inhibit anion transport. In N-ethylmaleimide-treated red cells, we have found that this binding site inhibits water transport and that the inhibition can be partially reversed by the specific stilbene anion exchange transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS), thus linking water transport to anion exchange. DTNB binding to this low-affinity site also inhibits ethylene glycol and methyl urea transport with the same KI as that for water inhibition, thus linking these transport systems to that for water and anions. These results support the view that band 3 is a principal constituent of the red cell aqueous channel, through which urea and ethylene glycol also enter the cell. PMID- 2992588 TI - Subcellular fractionation of pig coronary artery smooth muscle. AB - A detailed procedure for subcellular fractionation of the smooth muscle from pig coronary arteries based on dissection of the proper tissue, homogenization, differential centrifugation and sucrose density gradient centrifugation is described. A number of marker enzymes and Ca2+ uptake in presence or absence of oxalate, ruthenium red and azide were studied. The ATP-dependent oxalate independent azide- or ruthenium red-insensitive Ca2+ uptake, and the plasma membrane markers K+-activated ouabain-sensitive p-nitrophenylphosphatase, 5' nucleotidase and Mg2+-ATPase showed maximum enrichment in the F2 fraction (15-28% sucrose) which was also contaminated with the endoplasmic reticulum marker NADPH: cytochrome c reductase, and to a small extent with the inner mitochondrial marker cytochrome c reductase, and also showed a small degree of oxalate stimulation of the Ca2+ uptake. F3 fraction (28-40% sucrose) was maximally enriched in the ATP- and oxalate-dependent azide-insensitive Ca2+ uptake and the endoplasmic reticulum marker NADPH: cytochrome c reductase but was heavily contaminated with the plasma membrane and the inner mitochondrial markers. The mitochondrial fraction was enriched in cytochrome c oxidase and azide- or ruthenium red-sensitive ATP dependent Ca2+ uptake but was heavily contaminated with other membranes. Electron microscopy showed that F2 contained predominantly smooth surface vesicles and F3 contained smooth surface vesicles, rough endoplasmic reticulum and mitochondria. The ATP-dependent azide-insensitive oxalate-independent and oxalate-stimulated Ca2+ uptake comigrated with the plasma membrane and the endoplasmic reticulum markers, respectively, and were preferentially inhibited by digitonin and phosphatidylserine, respectively. This study establishes a basis for studies on receptor distribution and further Ca2+ uptake studies to understand the physiology of coronary artery vasodilation. PMID- 2992590 TI - Kinetics of (Na+ + K+)-ATPase: analysis of the influence of Na+ and K+ by steady state kinetics. AB - The influence of Na+ and K+ on the steady-state kinetics at 37 degrees C of (Na+ + K+)-ATPase was investigated. From an analysis of the dependence of slopes and intercepts (from double-reciprocal plots or from Hanes plots) of the primary data on Na+ and K+ concentrations a detailed model for the interaction of the cations with the individual steps in the mechanism may be inferred and a set of intrinsic (i.e. cation independent) rate constants and cation dissociation constants are obtained. A comparison of the rate constants with those obtained from an analogous analysis of Na+-ATPase kinetics (preceding paper) provides evidence that the ATP hydrolysis proceeds through a series of intermediates, all of which are kinetically different from those responsible for the Na+-ATPase activity. The complete model for the enzyme thus involves two distinct, but doubly connected, hydrolysis cycles. The model derived for (Na+ + K+)-ATPase has the following properties: The empty, substrate free, enzyme form is the K+-bound form E2K. Na+ (Kd = 9 mM) and MgATP (Kd = 0.48 mM), in that order, must be bound to it in order to effect K+ release. Thus Na+ and K+ are simultaneously present on the enzyme in part of the reaction cycle. Each enzyme unit has three equivalent and independent Na+ sites. K+ binding to high-affinity sites (Kd = 1.4 mM) on the presumed phosphorylated intermediate is preceded by release of Na+ from low-affinity sites (Kd = 430 mM). The stoichiometry is variable, and may be Na:K:ATP = 3:2:1. To the extent that the transport properties of the enzyme are reflected in the kinetic ATPase model, these properties are in accord with one of the models shown by Sachs ((1980) J. Physiol. 302, 219-240) to give a quantitative fit of transport data for red blood cells. PMID- 2992589 TI - Na+-Na+ exchange mediated by (Na+ + K+)-ATPase reconstituted into liposomes. Evaluation of pump stoichiometry and response to ATP and ADP. AB - (Na+ + K+)-ATPase from shark rectal glands reconstituted into lipid vesicles and oriented inside out catalyses an ouabain-sensitive Na+-Na+ exchange in the absence of intravesicular K+ when ATP is added extravesicularly. Intravesicular ouabain inhibited the exchange completely. This was also the case with digitoxigenin added to the vesicles. Intravesicular oligomycin inhibited the Na+ Na+ exchange partly in a fashion which was ATP dependent. The exchange is accompanied by a net hydrolysis of ATP with an apparent Km of 2.5 microM. ADP was found to give no stimulation of the Na+-Na+ exchange, contrarily, ADP inhibited the ATP-dependent exchange of Na+ both at optimal and supraoptimal ATP concentrations. When initial influx and efflux of 22Na was measured and the hydrolysis of ATP concomitantly determined a coupling ratio of 2.8:1.3:1 was found, i.e. 2.8 moles of Na+ were taken up (cellular efflux) and 1.3 moles of Na+ extruded (cellular influx) for each mole of ATP hydrolyzed. The electrogenic Na+ Na+ exchange generated a transmembrane potential which was measured with the fluorescent probe ANS (8-anilino-1-naphthalenesulfonic acid) to be 60 mV positive inside the liposomes (extracellular). PMID- 2992591 TI - Effects of ATP and cyclic AMP on the (Na+ + K+ + 2Cl-)-cotransport system in turkey erythrocytes. AB - As turkey erythrocytes were progressively depleted of ATP by preincubation with dinitrophenol, the (Na+ + K+ + 2Cl-)-cotransport system (assayed by the bumetanide-sensitive fraction of 86Rb+ influx) became less responsive to activation. The dependence upon intracellular ATP concentration was significantly steeper for transport activated by hypertonic shock (halfmaximal activity at 0.7 mM ATP) than for that activated by either epinephrine or cyclic AMP (halfmaximal activity at 1.7 mM ATP). Upon removal of epinephrine or cyclic AMP from cells that had been preincubated with those substances, bumetanide-sensitive transport activity declined sharply, even though the intracellular cyclic AMP concentration was still over 10-fold that required to maximally activate the transport system. These data are in agreement with the notion that the (Na+ + K+ + 2Cl-) cotransport system in turkey erythrocytes is activated by cyclic AMP, presumably through the 'classical' pathway involving a protein kinase. They do however indicate that some other, as yet undefined aspect of cyclic AMP metabolism is important for the maintenance of transport activity. PMID- 2992592 TI - Stability of [3H]ouabain binding to the (Na+ + K+)-ATPase solubilized with C12E8. AB - The (Na+ + K+)-ATPase from dog kidney and partially purified membranes from HK dog erythrocytes were labeled with [3H]ouabain, solubilized with C12E8 and analyzed by HPLC through a TSK-GEL G3000SW column in the presence of C12E8, Mg2+, HPO4(2-) and glycerol at 20-23 degrees C. The peaks of [3H]ouabain bound to the enzyme from dog kidney and HK dog erythrocyte membranes corresponded to each other with apparent molecular weights of 470 000-490 000. In addition, these bindings of [3H]ouabain to the (Na+ + K+)-ATPase were observed to be stable at 20 23 degrees C for at least 18 h after the solubilization. PMID- 2992593 TI - The prostasome: its secretion and function in man. AB - An intact organelle, the prostasome, is secreted by the acinar epithelial cell of the human prostate gland. The ultrastructural location of the prostasome is within membrane-bound storage vesicles in the epithelial cells. Prostasomes are delivered into the glandular lumen by an exocytotic event, which is preceded by fusion of adjacent membranes belonging to the storage vesicle and the epithelial cell. Alternatively, the storage vesicle can be translocated in toto from the cell interior into the acinar lumen through the plasma membrane. This latter event has been designated diacytosis. Both phenomena seem to occur with approximately equal frequency in the human prostate gland. An ATPase system that is Mg2+ and Ca2+-dependent is firmly linked to the membranes encasing the prostasomes. The ATPase system may be the molecular basis for vectorial transport of calcium into these organelles. Also a protein kinase activity is located in the membranes. An increase in membrane thickness was observed on phosphorylation. The physiologic function of the prostasomes is not known. They may be important for promoting forward motility of spermatozoa. PMID- 2992594 TI - Quantification of endocytosis-derived membrane traffic. AB - The main data covered by this article have been summarized in Table I. A fairly uniform picture is obtained for endocytosis-derived membrane transfer and compartmentation. This may be due to the limited amount of information and the resulting low resolution. Data on mainly three cell types are presented: macrophages, fibroblasts and amoebae. The data vary as much for one cell type as between different cells. Therefore, no possible differences related to cell function emerge. More detailed data, for more cell types, may change the picture. The values for cell surface area, although significantly different in absolute terms (column S in Table I), are rather similar when related to cell diameter, all being about 3-fold in excess of the surface area of the smooth sphere of comparable volume (column xi in Table I). The rate of plasma membrane internalization for macrophages and amoebae both professional phagocytes, is about 2 cell surface area equivalents per h or more. This may be somewhat higher than for fibroblasts (column PM/h in Table I). The average residence time for membrane on the cell surface, therefore, is about 30 min. A most interesting finding seems to be the rather uniform values obtained for the average size (volume weighted) of primary pinosomes, being about 0.3 micron in diameter (column phi-Internalization in Table I). Due to their rapid increase in size as a result of fusion (cf. Fig. 2), it has not been feasible to directly measure the size of primary pinosomes by morphometric means. The values in Table I, give no information on the size distributions of primary pinosomes and on whether these consist of one or more size classes. The steady-state average diameter of pinosomes is noticeably larger than that of primary pinosomes (column phi pinosomes in Table I; cf. Table II for Acanthamoebae). The corresponding decrease in surface-to-volume ratio can make about 50% of pinosomal membrane available for recycling directly from this membrane compartment. Membrane recycling from the pinosomal compartment occurs after an average residence time of about 3 min for macrophages and 4-6 min for fibroblasts (column tau-pinosomes in Table I). The relative pool size of intracellular membranes participating in shuttling to and from the cell surface is significantly different for animal cells and amoebae (column rho in Table I). For macrophages, fibroblasts, CHO cells, and mast cells, this intracellular membrane pool amounts to about 10-20% the plasma membrane area, compared to 150-200% in the case of amoebae.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2992595 TI - Purification and characterization of two distinct Ca2+-activated proteinases from hearts of hypertensive rats. AB - Two distinct Ca2+-activated proteinases were purified and characterized from hearts of hypertensive rats. Ca2+-activated proteinases I and II, having low and high Ca2+ requirements, respectively, were first separated by DEAE-cellulose chromatography. The enzymes were then purified individually by different column procedures: chromatography on phenyl-Sepharose, then Sephadex G-200 for proteinase I and reactive-red agarose for proteinase II. The apparent molecular weight of purified proteinase I was 125 000 and that for purified proteinase II was 110 000. Both enzymes are heterodimers made up of a larger catalytic subunit and a smaller subunit devoid of proteinase activity. Ca2+ concentrations for half maximal activation were 5 microM for proteinase I and 200 microM for proteinase II. Both enzymes were inhibited by sulfhydryl-modifying agents, but exhibited different characteristics in the auto-digestion reaction in the presence of Ca2+. Proteinases I and II were also purified from hearts of normotensive rats and shown to be identical to their respective counterparts from hearts of hypertensive rats. However, proteinase II activity in hypertensive rat hearts was significantly elevated as compared to controls. PMID- 2992596 TI - Complete co-purification of choline kinase and ethanolamine kinase from rat kidney and immunological evidence for both kinase activities residing on the same enzyme protein(s) in rat tissues. AB - Choline kinase and ethanolamine kinase were completely co-purified from rat kidney cytosol through acid treatment, ammonium sulfate fractionation, DEAE cellulose column chromatography, Sephadex G-150 gel filtration followed by choline-Sepharose affinity chromatography. The final preparation appeared to be highly homogeneous with respect to both native and sodium dodecyl sulfate polyacrylamide gel electrophoresis (Ishidate, K., Nakagomi, K. and Nakazawa, Y. (1984) J. Biol. Chem. 259, 14706-14710). Throughout the purification steps, the ratio of ethanolamine kinase activity to choline kinase activity was almost constant in a range of 0.3-0.4, which strongly indicated that, in rat kidney, both activities could reside on a single enzyme protein. The rabbit polyclonal antibody raised against highly purified rat kidney choline (ethanolamine) kinase protein inhibited both choline and ethanolamine kinase activities in a parallel manner in crude enzyme preparations not only from rat kidney, but also from rat liver, lung and intestinal cytosols. The results, together with our previous findings, suggested strongly that, in rat tissues, at least large portions of both kinase activities are present on the same enzyme protein(s). The kinetic properties of both kinase reactions with the highly purified kidney enzyme were compared in some detail and the overall result suggested that choline kinase and ethanolamine kinase activities may not have a common active site on a single enzyme protein. PMID- 2992597 TI - Bradykinin stimulates phospholipase C in rat renal medullary slices. AB - Rat renal medullary slices prelabeled with [14C]arachidonic acid generate [14C]diacylglycerol within 1 min of exposure to bradykinin action. Production of [14C]diacylglycerol is transient. 2 min after the addition of bradykinin, the levels of metabolite reach the maximum, but decrease thereafter. Simultaneously, bradykinin induces a parallel decrease of the radioactivity in phosphatidylinositol. No degradation of other phospholipids is observed, and triacylglycerol is not affected. The degradation of [14C]phosphatidylinositol to [14C]diacylglycerol indicated the presence of phospholipase C activity. Preincubation of prelabeled slices with 2 mM dibutyryl cyclic AMP prevents both the generation of diacylglycerol and the degradation of phosphatidylinositol. Neither mepacrine nor indomethacin block diacylglycerol production and phosphatidylinositol breakdown. We conclude that, when rat renal medullary slices are stimulated with bradykinin, phosphatidylinositol-specific phospholipase C is activated. PMID- 2992598 TI - Catabolism of leukotriene A4 into B4, C4, and D4 by rat liver subcellular fractions. AB - [3H]Leukotriene A4 was incubated with various subcellular fractions of rat liver homogenates. After solvent extraction and purification on C18 Sep-Pak cartridges, tritiated products migrating on reversed-phase HPLC with authentic unlabelled leukotriene C4, D4 and B4 were observed. The identity of leukotriene C4 was confirmed through enzymatic conversion into D4 by gamma-glutamyl transpeptidase as well as by bioassay on the rat stomach fundus after HPLC purification. The contractile response to the extracted material was blocked by the SRS antagonist, FPL 55712. Leukotriene B4 synthesis was located in the 100 000 X g supernatant, while C4 synthesis was present in the corresponding pellet. Leukotriene C4 formation was enhanced when reduced glutathione was supplemented in the incubation medium. These results demonstrate the presence in rat liver of various enzymatic steps in leukotriene A4 catabolism. PMID- 2992599 TI - Rates of cholesterol synthesis and low-density lipoprotein uptake in the adrenal glands of the rat, hamster and rabbit in vivo. AB - The absolute rate of cholesterol acquisition from de novo synthesis and from receptor-dependent and receptor-independent low-density lipoprotein (LDL) uptake was determined in the adrenal glands of the rat, hamster and rabbit under in vivo conditions. The rate of incorporation of [3H]water into cholesterol in the adrenal gland was much higher in the hamster (1727 nmol/h per g) and rabbit (853 nmol/h per g) than in the rat (71 nmol/h per g). Assuming that 23 atoms of 3H are incorporated into the cholesterol molecule during its biosynthesis, the absolute rates of cholesterol synthesis were then calculated to equal 59, 29 and 2.4 micrograms/h per g of adrenal gland in the hamster, rabbit and rat, respectively. Rates of LDL-cholesterol uptake were measured using a primed continuous infusion of [14C]sucrose-labeled homologous LDL (total LDL transport) and methylated human LDL (receptor-independent LDL transport). The rate of total LDL-cholesterol uptake in the adrenal gland was much higher in the rabbit (227 micrograms/h per g) than in the rat (18 micrograms/h per g) or hamster (6 micrograms/h per g). In all three species LDL uptake was mediated largely (greater than 93%) by receptor dependent mechanisms. In terms of total cholesterol acquisition, the hamster adrenal gland derived 10-times more cholesterol from de novo synthesis than from LDL uptake, whereas the converse was true in the rabbit. Rates of de novo synthesis and LDL-cholesterol uptake were both low in the rat adrenal gland, which is known to derive cholesterol mainly from circulating high-density lipoproteins. Thus, the adrenal gland acquires cholesterol for hormone synthesis from at least three different sources and the quantitative importance of these sources varies markedly in different animal species, including man. PMID- 2992600 TI - The effect of bezafibrate and long-chain fatty acids on peroxisomal activities in cultured rat hepatocytes. AB - Peroxisomal activities have been evaluated in cultured rat hepatocytes in the presence of bezafibrate or long-chain fatty acids added to the culture medium. All activities decreased continuously over a time period of 100 h in culture but selected activities were relatively increased as a function of the added effectors. This relative increase in peroxisomal activities was dose-dependent, discernible within the first 24 h in culture and consisted of activities related specifically to peroxisomal fatty acyl beta-oxidation, e.g., cyanide-insensitive palmitoyl-CoA oxidation, H2O2-forming palmitoyl-CoA oxidase and heat-labile enoyl CoA hydratase. Peroxisomal catalase or mitochondrial fatty acyl beta-oxidation (cyanide-sensitive) remained relatively unchanged. The relative increase in peroxisomal activities was accompanied by a respective increase in the number of peroxisomes as well as in thymidine incorporation rate. PMID- 2992602 TI - The interaction of reduced glutathione with active oxygen species generated by xanthine-oxidase-catalyzed metabolism of xanthine. AB - The interaction of reduced glutathione (GSH) with active oxygen species generated during xanthine-oxidase-catalyzed metabolism of xanthine was investigated. The only GSH-derived product detected in this system was oxidized glutathione (GSSG). Catalase inhibited the oxidation of GSH to GSSG by more than 80%, whereas superoxide dismutase exerted a smaller but significant inhibition of GSSG formation. Hydroxyl radical (OH) scavengers or desferrioxamine (1 mM) had no effect on GSSG formation. Using EPR spectroscopy and the spin trap 5,5 dimethylpyrroline-N-oxide (DMPO), the production of superoxide was observed by the detection of a DMPO-OOH radical adduct. This spectrum was altered by the inclusion of GSH (5 - 20 mM) in the reaction mixture, indicating the generation of a different radical species consistent with DMPO-glutathionyl radical adduct generation. PMID- 2992601 TI - Studies on enzyme-substrate interactions of cholinephosphotransferase from rat liver. AB - In order to elucidate the reaction mechanism and the substrate-binding sites, CDPcholine:1,2-diacylglycerol cholinephosphotransferase (EC 2.7.8.2), prepared from rat liver microsomal fraction, has been subjected to kinetic analysis and substrate specificity studies. Kinetic evidence supports the hypothesis of a Bi Bi sequential mechanism, involving a direct nucleophilic attack of diacylglycerol on CDPcholine during the reaction. To investigate the substrate requirements for recognition and catalysis, several CDPcholine analogs, modified in the nitrogen base or in the sugar or in the pyrophosphate bridge, have been synthesized, characterized and assayed as substrates and/or inhibitors of the reaction. The amino group on the pyrimidine ring, the 2'-alcoholic function of the ribose moiety as well as the pyrophosphate bridge have been identified as critical sites for enzyme-substrates interactions. PMID- 2992604 TI - Insertion of lipid domains into plasma membranes by fusion with erythrocytes. AB - After prelabeling the plasma membrane with several lipid-specific fluorescent probes, erythrocytes with symmetric lipid bilayers were fused with culture cells using either poly(ethylene glycol) or Sendai virus as fusogen. Several nonspecific probes were transferred to, and became uniformly distributed within, the culture cell membrane upon fusion. In contrast, when merocyanine 540, which displays preferential binding to bilayers in which the lipids are loosely packed, was used to prelabel erythrocytes, fluorescence remained localized within a small confined area of the membrane, even 24 h after fusion. These results suggest that insertion of the lipids of the erythrocyte membrane into the plasma membrane of the culture cell can produce discrete domains which persist as such for long periods following fusion. Because the inserted proteins of the erythrocyte membrane similarly do not freely diffuse throughout the culture cell membrane, interactions between membrane proteins and lipids may be involved in this singular compartmentalization. PMID- 2992605 TI - Casein kinases and their protein substrates in rat liver cytosol: evidence for their participation in multimolecular systems. AB - We have shown by gel filtration on Sepharose 4B at low ionic strength that casein kinases S (type 1), heparin-insensitive, and TS (type 2), heparin-inhibited, of rat liver cytosol participate in two distinct multimolecular systems, Ve/Vo = 1.25 and Ve/Vo = 1.90, respectively, both less retarded than the peak of cAMP dependent protein kinase activity (Ve/Vo = 2.04). Both casein kinase I and casein kinase II complexes are unstable in 0.5 M NaCl, giving rise by gel filtration under these conditions to the free forms of casein kinase S (Ve/Vo = 2.37, Mr 34 000) and casein kinase TS (Ve/Vo = 2.10, Mr 130 000), respectively. In contrast, the elution volume of cAMP-dependent protein kinase activity is always the same irrespective of the ionic strength of the medium. Casein kinase I, accounting for the whole casein kinase S activity of cytosol, also contains a phosphorylatable 31-kDa protein (p31) which is a substrate of casein kinase S, since its phosphorylation is insensitive to heparin, the heat-stable inhibitor and trifluoperazine, but it is prevented by beryllium. Casein kinase II, on the other hand, apparently results from the association of the whole casein kinase TS (type 2) of rat liver cytosol with a 90-kDa protein substrate (p90) which is distinct from glycogen synthase according to their different peptide mappings. The radiolabelling of p90 is inhibited by heparin, unlabeled GTP and polyglutamates, while it is dramatically and specifically enhanced by polylysine. At least three more protein bands of Mr 58 000, 52 000 and 37 000 are phosphorylated by casein kinase TS in the casein kinase II fraction: their co-elution with casein kinase TS, however, seems to be accidental and their radiolabeling in the presence of polylysine is almost negligible compared to that of p90. It is concluded that p31 and p90 may represent specific targets of casein kinase S and casein kinase TS, respectively, whose intimate association with the enzymes could be functionally significant. PMID- 2992603 TI - High concentrations of arachidonic acid induce platelet aggregation and serotonin release independent of prostaglandin endoperoxides and thromboxane A2. AB - We examined platelet aggregation and serotonin release, induced by less than 60 micro M arachidonic acid, using washed platelet suspensions in the absence of albumin. The concentration of arachidonic acid used did not cause platelet lysis. Platelet responses induced by less than 20 micro M arachidonic acid were inhibited by aspirin, whereas those induced by above 30 micro M arachidonic acid were not inhibited, even by both aspirin and 5,8,11,14-eicosatetraynoic acid. Although phosphatidic acid and 1,2-diacylglycerol increased after the addition of arachidonic acid in aspirin-treated platelets, the amounts were not parallel to platelet aggregation. Oleic, linoleic and linolenic acids also induced platelet responses, while palmitic, stearic and arachidic acids did not. EDTA, dibutyryl cyclic AMP, apyrase and creatine phosphate/creatine phosphokinase brought about almost the same effects in platelet responses induced by the unsaturated fatty acids, other than arachidonic acid, as those induced by 40 micro M arachidonic acid. These results suggest that the mechanism of the actions of more than 30 micro M arachidonic acid on platelets is the same as that of the other unsaturated fatty acids and is independent of prostaglandin endoperoxides, thromboxane A2 and, perhaps, phosphatidic acid and 1,2-diacylglycerol. PMID- 2992606 TI - Effect of thyroid hormone on intracellular Ca2+ mobilization by noradrenaline and vasopressin in relation to glycogenolysis in rat liver. AB - The relation between Ca2+ efflux, Ca2+ mobilization from mitochondria and glycogenolysis was studied in perfused euthyroid and hypothyroid rat livers stimulated by Ca2+-mobilizing hormones. Ca2+ efflux, induced by noradrenaline (1 microM) in the absence or presence of DL-propranolol (10 microM) from livers perfused with medium containing a low concentration of Ca2+ (approx. 24 microM), was decreased by more than 50% in hypothyroidism. This correlated with an equal decrease of the fractional mobilization of mitochondrial Ca2+, which could account for 65% of the difference between the net amounts of Ca2+ expelled from the euthyroid and hypothyroid livers. With vasopressin (10 nM) similar results were found, suggesting that hypothyroidism has a general effect on mobilization of internal Ca2+. In normal Ca2+ medium (1300 microM), however, the effect of vasopressin on net Ca2+ fluxes and phosphorylase activation was not impaired in hypothyroidism, indicating that Ca2+ mobilization from the mitochondria in this case plays a minor role in phosphorylase activation. The alpha 1-adrenergic responses of Ca2+ efflux, phosphorylase activation and glucose output, glucose-6 phosphatase activity and oxygen consumption in hypothyroid rat liver were completely restored by in vivo T3 injections (0.5 micrograms per 100 g body weight, daily during 3 days). Perfusion with T3 (100 pM) during 19 min did not influence hypothyroid rat liver oxygen consumption and alpha 1-receptor-mediated Ca2+ efflux. However, this in vitro T3 treatment showed a completely recovered alpha 1-adrenergic response of phosphorylase and a partly restored glucose-6 phosphatase activity and glucose output. The results indicate that thyroid hormones may control alpha 1-adrenergic stimulation of glycogenolysis by at least two mechanisms, i.e., a long-term action on Ca2+ mobilization, and a short-term action on separate stages of the glycogenolytic process. PMID- 2992608 TI - [Quantitative estimate of major factors determining the kinetics of electron transfer reactions with cytochrome c]. AB - Quantitative estimation of basic factors determining the electron transfer rate constant between cytochrome c and inorganic metal complexes and electron exchange rate constant based on the theory of nonadiabatic electron transfer in polar media is presented. PMID- 2992607 TI - Phosphatidylinositol hydrolysis and phosphatidylinositol 4',5-diphosphate hydrolysis are separable responses during secretagogue action in the rat pancreas. AB - Rat pancreatic fragments and acinar preparations were incubated in vitro to characterize further the changes in phosphoinositide metabolism that occur during secretagogue action. Two distinct responses were discernible. The first response, most notably involving a decrease in phosphatidylinositol content, was (a) observed at lower carbachol concentrations in dose-response studies, (b) inhibited by incubation in Ca2+-free media containing 1 mM EGTA, (c) associated with increases in inositol monophosphate production, and (d) provoked by all tissue secretagogues (carbachol, cholecystokinin, secretin, insulin, dibutyryl cAMP and the ionophore A23187), regardless of whether their mechanism of action primarily involved Ca2+ mobilization or cAMP generation. This decrease in phosphatidylinositol content was at least partly due to phospholipase C (and/or D) activation, as evidenced by the increase in inositol monophosphate. The second response, most notably involving markedly increased incorporation of 32PO4 into phosphatidic acid and phosphatidylinositol, was (a) observed at higher carbachol concentrations, (b) not influenced by incubation in Ca2+-free media containing 1 mM EGTA, and (c) associated with increases in inositol triphosphate production. This 32PO4 turnover response was probably largely the result of phospholipase C mediated hydrolysis of phosphatidylinositol 4',5'-diphosphate, which, as shown previously, also occurs at higher carbachol concentrations and is insensitive to comparable EGTA-induced Ca2+ deficiency. This phosphatidylinositol 4',5' diphosphate hydrolysis response was only observed in the action of agents (carbachol and cholecystokinin) which mobilize Ca2+ via activation of cell surface receptors. The present results indicate that phosphatidylinositol and phosphatidylinositol 4',5'-diphosphate hydrolysis are truly separable responses to secretagogues acting in the rat pancreas. Furthermore, phosphatidylinositol 4',5'-diphosphate, rather than phosphatidylinositol hydrolysis is more likely to be associated with receptor activation and Ca2+ mobilization. PMID- 2992609 TI - [Interaction of manganese II ions with proteins]. AB - Electron and ESR-spectroscopic study was carried out of the interaction between manganese(II) ions with albumine and lysozome in water solutions at different manganese concentrations and ratios manganese/protein. The results obtained suggest presence of metal-protein interactions. At high manganese concentrations liotropic effect is observed. At high molecular ratios protein/manganese and manganese concentrations close to physiological ones formation of coordination compounds manganes(II) ions with proteins was observed. PMID- 2992610 TI - [Intraneuronal information processing. Delay in the changes in membrane potential after administration of cyclic nucleotides]. AB - Depolarization of the neuron membrane induced by cyclic adenosine monophosphate (cAMP) was shown both by ionophoresis and by injection with pressure. Swelling of the neuron during the injection of various substances with pressure causes membrane depolarization which is similar to that induced by cAMP. When applying pressure the cAMP effect can be distinguished by introducing small volumes of concentrated solutions. Similarity between the effects of cAMP and mechanical stimulation of the neuron suggests that in both cases the effect involves action of the electromechanical system consisting of microskeleton and micromuscles which regulate permeability of molecular channels. The delay of the effect after the moment of cAMP and cGMP introduction is small, which enables a conclusion concerning their direct interaction with the cytoskeleton. PMID- 2992611 TI - Non-parametric statistics for nucleic acid sequence study. AB - The use of non-parametric statistics for nucleic acid sequence studies is illustrated by some examples. This method is highly flexible and allows design of specific tests for detecting sequence structure. Tests devoted to local repetitivity, codon nearest neighbors, and dinucleotide avoidance are discussed in detail. An appendix indicates all computations required to use these tests. PMID- 2992612 TI - The missense errors in protein can be controlled by selective synonymous codon usage at the level of transcription. AB - In the cases of the 6-fold degenerate residues and the stop signal, selective codon usage at the level of transcription can account for a 10-20% variation in their mistranslation rate. For all other residues, the mistranslation rate is dependent upon the degree of degeneracy only, but not upon the pattern of synonymous codon usage. PMID- 2992614 TI - [Study of the mechanism of initiation of enzymatic NADPH-dependent lipid peroxidation in membranes of the endoplasmic reticulum]. AB - The localization and mechanism of generation of active oxygen species in the enzymatic NADPH-dependent lipid peroxidation system in liver microsomes were studied. Using the spin-trapping method, the key role of active oxygen species in the initiation of NADPH-dependent enzymatic lipid peroxidation was confirmed. It was shown that active oxygen species are generated via consecutive one-electron reduction of the oxygen molecule by NADPH-cytochrome P-450 reductase. PMID- 2992615 TI - [Stabilization of synaptic membranes with alpha-tocopherol against the damaging effect of phospholipases. Possible mechanism of the biological action of vitamin E]. AB - Using fluorescent and EPR spin probing techniques, the effects of phospholipases A2, C and D on rat brain synaptosomal membranes were investigated. It was shown that treatment of synaptosomal membranes with phospholipases A2, C and D results in their depolarization and increase of their surface negative charge. In case of phospholipases A2 and C, these changes are also accompanied by a decrease of the microviscosity of the synaptosomal membrane lipid bilayer. alpha-Tocopherol protects synaptosomal membranes against the damaging action of phospholipases. The stabilization of synaptosomes by vitamin E consists in the reconstitution of the transmembrane potential and in an increased microviscosity of phospholipase treated membranes. The stabilizing effect of alpha-tocopherol is due to the binding of phospholipid hydrolysis products rather than to the inhibition of phospholipases. The observed stabilization of synaptosomal membranes by alpha tocopherol is interpreted as a feasible mechanism of biological effects of vitamin E on biological membranes. PMID- 2992613 TI - Information transmission by isolated frog muscle spindle. AB - Transmission of sensory information was calculated for the isolated frog muscle spindle receptor, using Shannon's information measure. Sinusoidal movements, random noise stretches, and sinusoids with superimposed auxiliary noise were applied as stimuli. In addition, the static prestretch level of the intrafusal muscle bundle was adjusted between resting length (L0) and L0 + 600 micron, so that the analysis of the information transmission properties covered the entire dynamic range of the sensory receptor organ. Sinusoidal stretches below 2 Hz evoked smoothly modulated cycle histograms, which were approximately linearly related to the stimulating sinewave. The transinformation rates under these conditions were generally low (5-17 bit X s-1), regardless of the amplitude of the applied movement. Increasing prestretch enhanced the modulation depth of the cycle histograms considerably, but increased the transinformation rates by less than 10 bit X s-1. By contrast, sinusoids above 2 Hz evoked clearly nonlinear cycle histograms, because each action potential was firmly phase-locked to a small segment of the stretch cycle. Under these conditions the transinformation rates grew larger with increasing stimulus frequency and approached 130 bit X s-1 at 60 Hz. Small amplitude sinusoidal stretches, however, evoked considerable transinformation rates in the high frequency region only then, when the spindle receptor was extended to higher prestretch levels. Random stretches evoked transinformation rates between 5 and 30 bit X s-1 depending on both the prestretch level and the intensity of the noise stimulus. The linear response components carried only about 25% of the transinformation rates transmitted by both the linear and nonlinear response components. Auxiliary noise stimuli greatly improved the information transmission of sinusoidal stretches. For example, a pure sinusoid evoked 5 bit X s-1. Adding a noise signal with equal energy to the sinusoidal movement elicited 20 bit X s-1. This facilitation effect of auxiliary noise was restricted to low frequency sinusoidal stimuli. The present results are discussed with respect to the information transmission properties of various sensory systems evaluated by either the same or different information processing procedure as that used in the present study. The functional significance of high transinformation rates sent by the muscle spindle to the central nervous system is discussed with respect to motor control. PMID- 2992616 TI - [Description of Na, K-ATPase activation by monovalent cations using a simplified mathematical model]. AB - A simple mathematic model describing the activation Na,K-ATPase system by univalent cations is proposed. The constants for the enzyme activation values by each of the ions in the presence of a fixed concentration of the other ion have been calculated. The substitution of these values into the common equation describing the behaviour of the whole system according to the given model gives the curve of Na,K-ATPase activity change in dependence of Na/K ration at the same total concentration 150 mM. The experimental points correspond to the curve. PMID- 2992617 TI - Cortisol response to vasopressin in depression. PMID- 2992618 TI - Triiodothyronine-induced reversal of learned helplessness in rats. PMID- 2992619 TI - Electron paramagnetic resonance and optical spectroscopic study of the Cu2+-tRNA system. AB - The interaction of copper ions with tRNA has been studied by optical and EPR spectroscopies. The interaction results in two different paramagnetic complexes characterized by a tetragonal symmetry of the ligand electric field sensed by the ions. The complete set of the spin Hamiltonian parameters has been extracted by computer simulation with the Monte Carlo method. Hypotheses concerning the putative ligands are put forward. PMID- 2992621 TI - Molecular-dynamics simulation of phenylalanine transfer RNA. II. Amplitudes, anisotropies, and anharmonicities of atomic motions. PMID- 2992620 TI - Electron spin resonance of electrochemically generated free radicals from diaziquone and its derivatives. AB - The one-electron electrochemical reduction of diaziquone (AZQ) and 12 analogs is analyzed using ESR spectroscopy and cyclic voltammetry. The hyperfine coupling constants arising from the interaction of the unpaired electron with the aziridine nitrogen nuclei fall within 1.20 and 2.26 G. Smaller couplings are observed arising from the protons and nitrogens in the carboethoxyamino groups. The in vitro activity of AZQ and its analogs is examined. Methyl groups in the aziridine rings increase the activity of some analogs. In the absence of aziridines, a chloroquinone compound with only carboethoxyamino groups was surprisingly active. This compound has a more positive cathodic peak than diaziquone. PMID- 2992622 TI - Heavy-atom effects on energy transfer from polynucleotides to terbium (III). PMID- 2992624 TI - [Is non-A non-B hepatitis connected to a retrovirus?]. PMID- 2992623 TI - Simulation of the diffusion-controlled reaction between superoxide and superoxide dismutase. I. Simple models. PMID- 2992625 TI - Antibodies and natural immunity. AB - Natural antibody has been proposed as a mediator of T-independent (natural) resistance to tumors and microbial pathogens. In this article, it is argued that the "priming" of the B cell repertoire by ubiquitous microorganisms and autoantigens results in natural antibody production and this response offers a survival advantage for the host as a consequence of the participation of these antibodies in natural resistance. PMID- 2992626 TI - Angiotensin as a model for hormone--receptor interactions. AB - Proton magnetic resonance and chemical reactivity studies have demonstrated the presence of a tyrosine charge relay system in angiotensin which is analogous to the serine charge relay system present at the active site of serine proteases. Receptor activation by angiotensin can be explained by electronic effects deriving from an interaction of the charge relay system with stacking of the histidine and phenylalanine rings. Experiments with serine protease inhibitors suggest the possibility that mechanistic features of the interaction of angiotensin with its receptors may apply to other 'phenoxyl' hormones including certain peptides, steroids and catecholamines. PMID- 2992627 TI - [Effect of ACTH on the corticosteroid and progesterone levels in female hamadryas baboons in relation to the menstrual cycle phase]. AB - The effect of exogenous ACTH on the endogenous function of the steroid-producing glands has been investigated in hamadryas baboon females depending on the level of sex hormones in the body. The content of corticosteroids and progesterone in peripheral blood plasma was measured by radioimmunoassay. It has been established that the time course of changes in the adrenal gland response to administration of a standard ACTH dose was of the same type in the different menstrual cycle phases. The inhibitory effect of ACTH on progesterone secretion in ovarian corpus luteum being active has been revealed. PMID- 2992628 TI - [ACTH, corticosterone and beta-endorphin in the blood plasma of rats under prolonged immobilization stress]. AB - Radioimmunoassays were employed to study variation in the concentration of corticosterone, ACTH and beta-endorphin in rat blood plasma at different times of the 30-hour immobilization and under the effect of a short-term action of ecologically significant negative stimulation emanating from other animals. Both prolonged immobilization stress and short-term emotional reaction produced appreciable alterations in the blood plasma content of all the hormones under study. The findings indicate that variations in the corticosterone and beta endorphin levels were in the best agreement. PMID- 2992629 TI - [Transmembrane potential and electrogenicity of thrombocyte Na, K-ATPase in different forms of hypertension in rats]. AB - The fluorescent dye 3,3'-dipropylthiadicarbocyanine iodide, dis-C3-(5), was used to study the membrane potential of platelets in rats with renal, DOCA-induced and spontaneous hypertension. The magnitude of the transmembrane potential in platelets combines the activity of electrogenic Na, K-ATPase and the rate of ion diffusion across the plasma membrane. The basal potential is 12 = 14 mB as lowered only in spontaneously hypertensive rats. The decrease of the basal potential is made for electrogenicity of platelet Na, K-ATPase in spontaneously hypertensive rats. PMID- 2992630 TI - [Combined action of lithium salts and serotonin on the caudate nucleus]. AB - Intracaudate injections of lithium hydroxybutyrate (5 micrograms) produce a serotonin-like effect on the power characteristics of the caudatogram frequency ranges. However, when injected in a single dose, the drug is a serotonin antagonist at the level of the caudate nucleus. On the contrary, being administered for a long time, the drug potentiates the action of the mediator. PMID- 2992631 TI - [Isolation of substances inhibiting the specific binding of imipramine and serotonin uptake from the brain of the bull]. AB - Several fractions that inhibit specific binding of 3H-imipramine and reverse uptake of 3H-serotonin in rat brain synaptosomes were obtained by gel chromatography on Sephadex G-10 from an acidic extract of brain homogenates. The profiles of inhibition of 3H-imipramine specific binding and reverse uptake of 3H serotonin were found to be in a good agreement. This finding suggests that substances identified in the brain extract may have both types of activities. Further investigations relative to the purification and identification of the substances isolated from bovine brain should be aimed at the search for endogenous ligands for the "imipramine receptor". PMID- 2992632 TI - [Role opioidergic and adrenergic mechanisms in realizing the pain-alleviating action of clofelin]. AB - It has been demonstrated on unrestrained rats that the analgesic effect of clofelin is primarily determined by the influences on the processes of adrenergic but not opioidergic mediation. The functional heterogeneity of receptor substrates involved in the realization of the hemodynamic (alpha 2-) and emotional-behavioral (alpha 1-) components of the analgesic effect of clofelin is suggested. PMID- 2992634 TI - [Identification of the number of mutant copies of the genetic transfer factor AP42]. AB - The copy number mutant of genetic transfer factor pAP42 (incFIX) was induced by the treatment of E. coli AP115 cells carrying the plasmid by N-methyl-N-nitro-N nitrosoguanidine. The copy number mutant pAP42::Tn1cop1 is characterized by the increased copy number. The cells carrying the copy number mutant have a higher ampicillin resistance and higher beta-lactamase activity. Mutant plasmid pAP42: :Tn1cop1 is incompatible with plasmid pAP42 and compatible with plasmids of the other inc-groups of F-like plasmids. PMID- 2992633 TI - [Action of alcohol in chronic emotional pain stress in rats]. AB - Male albino rats were exposed to daily emotional painful stress (EPS) for 4 weeks. The arterial blood pressure of the stressed animals increased and the dynamics of the heart rate changed after functional loading (hypokinesis during one or two hours) as compared with the control group. The increase of the heart weight and activation of cytochrome oxidase activity in the cerebral cortex and hippocampus of rats exposed to EPS were also demonstrated. The use of 20% ethyl alcohol instead of drinking water during EPS partially prevented vegetative disturbances and activation of hippocampal cytochrome oxidase and fully prevented the heart hypertrophy and activation of the enzyme in the cortex. Alcoholization resulted in the increased weight of the spleen and brown adipose tissue and thymus involution. A possible mechanism of the antistress action of alcohol linked with normalization of intensified lipid peroxidation under stress is discussed. PMID- 2992635 TI - [Systems regulating the tra genes of derepressed F-like plasmids]. AB - A study was made of the ability of reference plasmids of the 6 known Fin-groups to inhibit the functions of transfer genes (tra-genes) of the 4 derepressed F like plasmids (pAP22-2, pAP38, pAP43, pAP53). It was shown that unlike the derepressed Flac plasmid, the conjugation transfer of pAP38 and pAP53 plasmids was inhibited only by, the FinV plasmid, whereas pAP22-2 plasmids by Fin V and Fin V plasmids. The formation of donor-specific pili in case of pAP38 plasmid was inhibited by Fin Q, Fin U and Fin V plasmids, in case of pAP43 plasmid by Fin U Fin V and Fin W plasmids. PMID- 2992636 TI - The concentration of IgG in the serum is a major determinant of Fc-dependent reticuloendothelial function. AB - Defective Fc receptor-specific reticuloendothelial (RE) function has been reported in certain patients with a variety of immunologic and nonimmunologic diseases. The mechanism responsible for the impaired RE function is uncertain, but it could be caused by immune complexes that are present in many of these disorders. Alternatively, the impaired RE function could be a secondary effect of the high concentrations of monomeric IgG in the serum of these patients, since monomeric IgG can compete with complexed IgG for macrophage receptors in vitro. We studied the Fc-dependent RE function in 30 healthy control subjects and 27 patients using IgG-coated radiolabeled autologous red cells. There was a significant relationship between the concentration of IgG in the serum and the rate of clearance of antibody-sensitized cells (r = 0.51, P less than .01). Patients with hypergammaglobulinemia had the slowest Fc-dependent clearance, whereas those with hypogammaglobulinemia had the most rapid clearance. Immune complexes (Raji or polyethylene glycol) could not be shown to contribute to Fc dependent RE clearance above the effect of the IgG in the serum. The unusually rapid clearance in a patient with hypogammaglobulinemia could be returned to normal by raising the concentration of IgG in the serum. This study supports the concept that serum (monomeric) IgG competes with immune complexed IgG for macrophage Fc receptors in vivo. The competition for Fc receptors determines the level of competence of Fc-dependent RE function. Based on the results of this study, one can predict that a number of disorders characterized by hypergammaglobulinemia also will have impaired Fc-dependent RE function. PMID- 2992637 TI - Interleukin 2 (IL 2) and interferon-gamma production by T lymphocytes from patients with B-chronic lymphocytic leukemia: evidence that normally released IL 2 is absorbed by the neoplastic B cell population. AB - The capacity of T lymphocytes from patients with B cell chronic lymphocytic leukemia (B-CLL) to release interleukin 2 (IL 2) and interferon (IFN)-gamma was assessed following various stimuli. The spontaneous release of IL 2 and IFN-gamma was practically absent both with B-CLL and normal T lymphocytes. By contrast, after stimulation with phytohemagglutinin (PHA) or with PHA plus 12-O tetradecanoylphorbol-13-acetate, the production of IL 2 and IFN-gamma by B-CLL T lymphocytes was similar to that of normal T lymphocytes, irrespective of the reversed T lymphocyte subset distribution (OKT4/OKT8 ratio) observed in B-CLL. However, the titer of IL 2 was greatly reduced when autologous leukemic B cells were added to the culture system. Unlike IL 2, the presence of leukemic B cells did not affect the titer of IFN-gamma in the culture supernatants. The indication that IL 2 may be adsorbed in vivo by the neoplastic B cells was further confirmed by the demonstration of the IL 2 receptor (revealed by anti-Tac monoclonal antibody) on the leukemic B cells, particularly following mitogenic stimulation, and by the evidence that exogenous IL 2 can be directly absorbed by untreated B CLL T lymphocytes to release IFN-gamma and IL 2 is preserved, but that IL 2 may be rapidly removed by the neoplastic B-CLL cells, thus contributing to the well documented T lymphocyte abnormalities present in this disease. PMID- 2992638 TI - Induction and enhancement by monocytes of antibody-induced modulation of a variety of human lymphoid cell surface antigens. AB - We have previously reported that the addition of monocytes results in enhanced modulation of the T65 antigen when normal or leukemic lymphoid cells were cultured in vitro with the T101 monoclonal antibody. In the present investigation, we extend these findings to demonstrate that monocyte-enhanced modulation is a phenomenon that occurs with a variety of T and B lymphoid antigens identified by murine monoclonal antibodies. Two patterns of monocyte enhanced modulation were observed: (1) augmentation by monocytes of existing antigen modulation by the T101 and anti-Leu-4 antibodies, and (2) induction by monocytes of previously unrecognized modulation with the anti-Leu-2 and anti-Leu 9 antibodies. Enhancement of modulation by monocytes was also detected with antibodies to surface IgM and HLA-DR antigens. Antigen modulation on lymphoid cell lines appeared to be more variable than on fresh cells, with or without monocytes. Monocyte-enhanced antigen modulation was not demonstrated with two monoclonal antibodies against solid tumors. Monocyte-enhanced modulation was shown to be dependent upon the Fc portion of the antibody, but independent of proteolytic or oxidative compounds released by monocytes. These findings indicate that the results obtained during in vitro studies of antigen modulation may vary with the source of cells and the extent to which monocytic cells are present. In addition, these findings suggest an enhanced role for Fc receptor-bearing cells of monocytic origin in antigen modulation following in vivo administration of monoclonal antibodies. PMID- 2992639 TI - Purification and characterization of the human neutrophil NADH-cytochrome b5 reductase. AB - NADH-cytochrome b5 reductase is the predominant NADH-diaphorase found in the human neutrophil (Blood 62:152, 1983). Although this reductase segregates with the light membranes of nitrogen-cavitated neutrophils separated on Percoll gradients (which include the plasma membrane markers alkaline phosphatase and NADPH-oxidase), it is approximately 95% excluded from plasma membrane-enriched phagocytic vacuoles. The reductase constitutes approximately 5% of the light membrane fraction FAD-flavoprotein (14.8 +/- 5.5 pmol/mg protein) and was found in equimolar concentration with a high potential b cytochrome also present in this light membrane fraction and tentatively identified as cytochrome b5. Isolation of the reductase from human neutrophils was accomplished by Triton X 114 solubilization of the light Percoll gradient membranes, followed by temperature-dependent phase separation and then affinity chromatography on AMP Sepharose. The active preparation contained 1.3 mol FAD/mol protein, migrated on sodium dodecyl sulfate-polyacrylamide gels as a single band corresponding to an apparent mol wt of 45,000 daltons, exhibited a pl of 5.7 on chromatofocusing and was obtained in greater than 70% yield, with an overall purification of almost 900-fold. The purified enzyme was characterized by a high specificity for NADH as electron donor (Km = 6.4 mumol/L v Km greater than 1.6 mmol/L for NADPH) and exhibited a maximal turnover of ca. 30,000 min-1 at 22 degrees C with either ferricyanide or cytochrome b5 (Km = 10 nmol/L) as electron acceptor. Although the physical characterization and biochemical properties described here demonstrate that this neutrophil NADH b5 reductase is similar to the corresponding liver and erythrocyte enzymes, its unique function in the neutrophil has yet to be determined. PMID- 2992640 TI - HTLV-III/LAV antibody status of spouses and household contacts assisting in home infusion of hemophilia patients. AB - Thirty-four adult and pediatric hemophilia A and B patients and 50 nonhemophilic members belonging to 28 families were enrolled in August 1984 in a study of human T cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV III/LAV) antibody status and T cell subpopulation numbers. All 50 household contacts, including three spouses of LAV antibody-positive adult hemophiliacs, were immunologically normal and serologically negative with respect to HTLV III/LAV. Based on Western blot serologic testing of blood samples collected intermittently between July 1981 and August 1984 from 33 representative St Louis hemophiliacs studied during the period from 1981 to 1984, the average time since seroconversion was estimated as 20 months. One spouse of a seropositive hemophiliac and 23 parents of 27 seropositive pediatric hemophiliacs assisted regularly with home infusions. These infusion assistants have collectively experienced 44 person-years of concentrate infusion "exposure" without seroconversion. These results suggest that the likelihood for transmission of HTLV-III/LAV from hemophiliacs to persons assisting in their therapy is extremely low. PMID- 2992641 TI - Effects of dibutyryl adenosine 3',5'-cyclic monophosphate on erythropoietin production in human renal carcinoma cell cultures. AB - A human renal carcinoma from a patient with erythrocytosis, serially transplanted into athymic nude mice, was grown in primary monolayer cell cultures. After reaching confluency, the cultured cells formed multicellular hemicysts (domes), which became more abundant as the cultures approached saturation density. Erythropoietin (Ep) production by this renal carcinoma in culture was only slightly increased at the time of semiconfluency but showed a marked increase after the cultures reached confluency, in parallel with dome formation. Dibutyryl adenosine 3',5'-cyclic monophosphate significantly (P less than .01) stimulated Ep production and dome formation in the semiconfluent and confluent cultures of the renal carcinoma. PMID- 2992642 TI - Lymphadenopathy-associated virus antibodies and T cells in hemophiliacs treated with cryoprecipitate or concentrate. AB - Evidence for exposure to lymphadenopathy-associated virus (LAV) was investigated in 48 patients with hemophilia, 15 of whom had been treated exclusively with single-donor cryoprecipitate. The prevalence of antibodies to LAV in all patients was 53% in 1983 and 63% in 1984, while in patients treated only with cryoprecipitate, the prevalence was 31% in 1983 and 40% in 1984. Patients treated with any concentrate had a seroprevalence of 65% in 1983 and 77% in 1984. Seropositive patients were more likely to have a significant reduction in the ratio of helper to suppressor T cells, absolute numbers of helper T cells, and T cell function in vitro. Seven of 18 patients who were seronegative in 1983 had seroconverted by 1984. The relative risk of seroconversion for patients using any concentrate since 1981 compared with those using cryoprecipitate only was 3.9 (P = .04). Nevertheless, the rate of conversion in the latter group was 18% per year. PMID- 2992643 TI - Hemophilia B with inhibitor: molecular analysis of the subtotal deletion of the factor IX gene. AB - The structure of factor IX gene was analyzed in a hemophilia B patient with inhibitor. Genomic DNA, digested with a variety of restriction endonucleases, was hybridized with the cDNA and various genomic factor IX probes. A large subtotal deletion of the gene was observed. The borders of the deletion span from a approximately 125 nucleotide region within the last exon to an unknown domain at least 7.5 kb upstream from the first exon: it thus involves approximately 33 kb of the factor IX locus. The abnormal gene was inherited by the daughter of the propositus, who showed both the normal and the deleted allele. PMID- 2992644 TI - Antibody seronegative human T-lymphotropic virus type III (HTLV-III)-infected patients with acquired immunodeficiency syndrome or related disorders. AB - The human T-lymphotropic virus type III (HTLV-III) is the primary cause of the acquired immunodeficiency syndrome (AIDS) and related disorders (ARC). Prior studies have reported that nearly all symptomatic patients with AIDS or ARC manifest antibody to HTLV-III. This observation has engendered efforts to screen for HTLV-III, especially prior to blood donation, with assays for antibody to HTLV-III. We report the first two cases, one with AIDS and one with ARC, that are HTLV-III virus positive but antibody negative. Accurate diagnosis of HTLV-III infection in some cases may require direct virus culture or tests for antigen. In addition, lack of HTLV-III antibody may indicate an atypical clinical course of AIDS. PMID- 2992645 TI - Natural antibodies in sera from Japanese individuals infected with HTLV-I do not recognize HTLV-III. AB - Seventy-one sera from Japanese individuals infected with human T cell leukemia virus type I (HTLV-I) were examined for the presence of antibodies to HTLV-III by an enzyme-linked immunosorbent assay (ELISA) and by a strip radioimmunoassay based on the Western blot technique. The sera were from 23 healthy carriers and from 48 patients, including 18 with smoldering adult T cell leukemia (ATL), 13 with chronic ATL, and 17 with acute ATL. All people tested lived in the southwestern part of Japan, a known endemic area for HTLV-I infection. Antibodies against HTLV-I were detected in all sera both by indirect immunofluorescent methods and strip radioimmunoassay using cell lysates. Six sera were reactive in the ELISA assay for HTLV-III. But these sera did not react specifically to HTLV III-related proteins (p15, p24, gp41) when analyzed by strip radioimmunoassay. Our data suggest that coincidental infection of HTLV-I and HTLV-III is quite rare in Japan. PMID- 2992646 TI - A monoclonal antibody D51 recognizes the transferrin-receptor structure. AB - A monoclonal antibody D51 has been obtained during immunization against human fetal thymus. The antibody binds to a variety of leukemic cells. This is similar to the staining pattern of the mAb OKT9 and L 5.1, which recognize the transferrin receptor. However, there are some differences. The antibody immunoprecipitates a protein present on Molt-4 cells. Under reducing conditions this protein has a molecular weight of 90 K dalton. Under non-reducing conditions it has a molecular weight of 180 K dalton. This suggests a dimeric structure. The consecutive immunoprecipitation with D51 and OKT9, respectively, demonstrates that both antibodies recognize the structure of the transferrin receptor. PMID- 2992647 TI - Monoclonal antibody DF3 correlates with tumor differentiation and hormone receptor status in breast cancer patients. AB - The murine monoclonal antibody (MAb) designated DF3, reacts with a 300-kd human mammary epithelial antigen which is expressed on apical borders of secretory mammary epithelial cells and in the cytosol of less differentiated malignant cells. Human mammary tumors have been evaluated for the level of DF3 antigen as a correlate to clinicopathologic parameters related to degree of tumor differentiation: nuclear grade (NG), histologic grade (HG), and estrogen receptor status (ER). More DF3 antigen was present in breast carcinomas with NG 1 and 2 as compared to tumors with NG 3 (p = .002). Similarly DF3 antigen presence was greater in HG 1 and 2 tumors than in HG 3 (p less than .001). The results also demonstrate that quantitative differences in the presence of the DF3 differentiation antigen correlate with estrogen receptor status. Twenty-two of 23 ER positive tumors were also DF3 positive. Only 6 of 23 ER negative tumors were reactive to MAb DF3 (p less than .001). There was, however, no correlation between DF3 reactivity and absolute levels of estrogen or progesterone receptor. These findings confirm our hypothesis that MAb DF3 reacts to a differentiation antigen present in some human breast carcinomas. The DF3 antigen phenotype can serve as an independent phenotypic marker with correlations to standard indicators of degree of differentiation and estrogen receptor status of infiltrating ductal carcinomas of the breast, and should thus be evaluated as a prognostic indicator in breast cancer patients. The data also suggests that DF3 histochemistry may be a useful alternative in assessing estrogen receptor status of small breast cancers where there is an insufficient amount of tumor present for biochemical assay of hormone receptor levels. PMID- 2992648 TI - Clonidine and presynaptic adrenoceptor theory. AB - The effects of clonidine, a presumed selective presynaptic alpha 2-adrenoceptor agonist or partial agonist, were examined in guinea-pig atria. Split left atrial preparations were stimulated transmurally at 2 Hz with 100 pulses of 0.5 ms duration and the efflux of 3H-transmitter determined. Clonidine inhibited efflux at 3 X 10(-8)M to 3 X 10(-7)M by about 30%. Yohimbine, at a concentration (10( 6)M) which caused a 3 fold increase in the release of 3H-transmitter during field stimulation, did not alter the ability of clonidine to inhibit transmitter efflux. At 10(-6)M clonidine alone had no significant effect on the stimulation induced efflux of 3H-transmitter, but in the presence of yohimbine (10(-6)M), inhibited efflux by over 50%. The inhibitory effect of noradrenaline (10(-6)M) on 3H-transmitter efflux was antagonized by clonidine at 10(-6)M but not at 10(-8)M, although neither concentration of clonidine alone inhibited transmitter efflux. The present findings indicate that the effects of clonidine on the efflux of noradrenaline from sympathetic nerves cannot be accommodated within the currently held view that the compound is an agonist or partial agonist on presynaptic alpha 2-adrenoceptors. It appears that clonidine has multiple sites of action few of which are antagonized by a concentration of the prototypical presynaptic antagonist yohimbine, which enhances efflux 3 fold. PMID- 2992649 TI - Forskolin: its effects on potassium-evoked release of vasopressin from the rat neurohypophysis. AB - The effect of forskolin, added either before or 5 min after the onset of potassium-evoked release of vasopressin from isolated neurointermediate lobes of the rat has been investigated. A low concentration of forskolin (1 microM), added before stimulation, enhanced the potassium-evoked release of vasopressin throughout two successive 5 min periods of stimulation. Higher concentrations of forskolin (10-80 microM) produced no effect on the potassium-evoked release of hormone during the first 5 min of stimulation, but inhibited release during the second 5 min period. When added 5 min after the onset of potassium stimulation, forskolin (1-80 microM) reduced the amount of vasopressin released during the remaining 5 min of stimulation. Forskolin produced a concentration-dependent increase in cyclic AMP during both the control and potassium stimulation periods. The amount of cyclic AMP generated by forskolin during potassium stimulation was less than that produced during the corresponding control periods. PMID- 2992650 TI - Desensitization of prostacyclin responsiveness in a neuronal hybrid cell line: selective loss of high affinity receptors. AB - The binding of [3H]-iloprost (ZK36374) to NCB-20 membranes revealed a single population of high affinity receptors (KD = 9.55 nM, Bmax = 431 fmol mg-1 protein) and a low affinity, non-saturable binding component. Desensitization of prostacyclin-responsiveness of NCB-20 cells is induced by culture in the presence of the stable prostacyclin analogue carbacyclin. Desensitization is accompanied by an increase in the Kact value for prostacyclin (64.1 nM to 175 nM), and a reduction in the prostacyclin-dependent increase in adenylate cyclase activity (41.2 to 15.1 pmol cyclic AMP min-1 mg-1 protein). Desensitization is not accompanied by changes in the coupling of the catalytic (C) to the regulatory (Ns) subunit of adenylate cyclase. In addition, the physical identity of the receptor molecule (as characterized by its sensitivity to electron bombardment in the beam of a linear accelerator) is not changed by desensitization. Desensitization of prostacyclin-dependent activation of adenylate cyclase may be explained most simply by a loss of prostacyclin receptors. The anomalous increase in the Kact (concentration of prostaglandin giving half-maximum enzyme activation) for prostacyclin-stimulated adenylate cyclase was not accompanied by a substantial change in the KD of [3H]-iloprost binding, and is explained by a loss of spare receptors. Prostacyclin responsiveness in non-dividing cells may be restored after desensitization by prolonged culture (up to 48 h) in the absence of carbacyclin. Resensitization is accompanied by restoration of the high affinity Kact value (143 nM to 45.5 nM), and is dependent on de novo protein synthesis. PMID- 2992651 TI - Similarity between mu-opioid receptors in mouse vas deferens and guinea-pig ileum. AB - The effects of the opioid receptor agonist RX783006 and of the opioid receptor partial agonist (+)-meptazinol have been examined on electrically-induced twitch responses of the guinea-pig isolated ileum and of the mouse isolated vas deferens. Log10 concentration-tissue state curves were determined for (+) meptazinol and for RX783006, alone, in combination and, when appropriate, in the presence of naloxone (30 nM). Analysis of these log10 concentration-tissue state curves using the null equations derived and verified in the previous paper allows quantitation of the characteristics of the interaction of (+)-meptazinol with the opioid receptors in these tissues. The results indicate that the apparent differences in the actions of (+)-meptazinol on isolated electrically-stimulated guinea-pig ileum and mouse vas deferens can be accounted for without the need to postulate differences between mu-opioid receptors in these two tissues. PMID- 2992652 TI - Interaction of prostaglandin D2 with prostacyclin, carbacyclin and the hydantoin prostaglandin, BW245C, in guinea-pig platelets. AB - The anti-aggregating actions of prostaglandin D2 (PGD2) have been compared to prostacyclin (PGI2), its stable analogue carbacyclin and a hydantoin prostaglandin, BW245C, in guinea-pig PGI2, carbacyclin and BW245C were potent inhibitors of ADP-induced aggregation in guinea-pig platelets, with ID50 values comparable to those obtained in human platelet-rich-plasma. In contrast, PGD2 acted as a weak and partial inhibitor in guinea-pig platelet aggregation, producing a bell-shaped dose-response relationship. PGD2 induced a dose-related antagonism of the inhibitory actions of BW245C, prostacyclin and carbacyclin on guinea-pig platelets. However, PGD2 did not antagonize the inhibitory actions of either forskolin or dibutyryl cyclic AMP on this platelet preparation. The results suggest a non-specific interaction of PGD2 with these prostanoid binding sites on guinea-pig platelets. PMID- 2992653 TI - PACPX--a substituted xanthine--antagonizes both the A1 and A2 subclasses of the P1-purinoceptor: antagonism of the A2 subclass is competitive but antagonism of the A1 subclass is not. AB - 1,3-Dipropyl-8-(2-amino-4-chlorophenyl)xanthine (PACPX) was examined for its ability to antagonize adenosine acting on the A1 and A2 subclasses of the P1 purinoceptor. A1-purinoceptors were studied in the isolated, driven left atria of the guinea-pig, and A2-purinoceptors in the isolated, carbachol-contracted taenia coli of the guinea-pig. PACPX antagonized the actions of adenosine in both types of preparation and was a more potent antagonist than 8-phenyltheophylline. The antagonism at the A2-purinoceptor was competitive with a pA2 of 5.95. The antagonism at the A1-purinoceptor was not competitive, although antagonism at the A1-purinoceptor was greater than that at the A2-purinoceptor, based on a comparison of pD2 values. The manner of antagonism of PACPX on the A1 purinoceptors of the heart was different from that found for the A1-receptors in bovine brain, implying that there is a fundamental difference between these central and peripheral A1 subclasses of P1-purinoceptor. PMID- 2992654 TI - A microiontophoretic study of the actions of the putative sleep factor, piperidine, in the rat brainstem. AB - By means of microiontophoresis, we have compared the actions of a putative sleep substance, piperidine, with other neurotransmitters in the rat anaesthetized with urethane. In the pons and midbrain, piperidine mimicked the actions of acetylcholine on more than 200 neurones. Piperidine- and acetylcholine-induced excitations were equally effectively antagonized by hexamethonium or atropine. In 32 neurones piperidine showed no affinity for the receptors for the excitatory amino acid agonists, quisqualate and N-methyl-D-aspartate, piperidine-evoked excitations being unaffected by the antagonists glutamate diethylester or 2-amino 5-phosphonovalerate. Similarly, piperidine-evoked excitations in 23 neurones were unaffected by alpha-methylnoradrenaline, suggesting that piperidine does not act at receptors for noradrenaline. Twenty per cent of neurones responsive to piperidine were inhibited. These inhibitions in 12 neurones were insensitive to either strychnine or bicuculline indicating that piperidine does not act on receptors for glycine or gamma-aminobutyric acid. In a further 68 neurones, neither hexamethonium (4 out of 59 cells) nor atropine (0 out of 9 cells) was effective in antagonizing the inhibitions evoked by piperidine or by acetylcholine. It is suggested that piperidine may exert its central hypnogenic effects by an action at cholinoceptors in brainstem areas involved in sleep regulation. PMID- 2992655 TI - New models for the evaluation of opioid effects in the guinea-pig ileum. AB - The pharmacology of morphine and opioid peptides was studied in the guinea-pig ileum by examining their inhibitory effects on propulsive peristaltic activity and on the cooling-induced longitudinal contraction. In these experiments, dose response curves were recorded. The rank order of potency in inhibiting peristalsis was found to be: dermorphin greater than FK 33-824 greater than dynorphin-(1-17) greater than dynorphin-(1-13) greater than delta-receptor peptide greater than morphine greater than [Leu] enkephalin, whereas the rank order in inhibiting cooling-induced contractions was found to be: dynorphin-(1 13) congruent to FK 33-824 congruent to dermorphin greater than delta-receptor peptide greater than morphine. Naloxone antagonized the maximally effective dose of each of the opioid agents. In view of the differences between the abilities of these opioids to inhibit propulsive peristaltic activity, these models seem to be valuable for the examination of inhibitory opioid effects in the gut. PMID- 2992656 TI - The effects of milrinone and piroximone on intracellular calcium handling in working myocardium from the ferret. AB - The effects of milrinone and piroximone were compared to those of isoprenaline, dibutyryl adenosine 3':5'-cyclic monophosphate (dibutyryl cyclic AMP), forskolin, isobutylmethylxanthine, increased extracellular calcium [( Ca2+]o) and caffeine in ferret right ventricular papillary muscles that were loaded intracellularly with aequorin, a bioluminescent calcium indicator that emits light when it combines with calcium. The positive inotropic action of each drug, except caffeine, was associated with an increase in the peak amplitude of the aequorin light signal (i.e. intracellular Ca2+ transient) reflecting an increased amount of calcium available for excitation-contraction coupling; the positive inotropic effect of caffeine appears to occur by other mechanisms. The time courses of the aequorin light signal and corresponding tension response were shortened by isoprenaline, forskolin, isobutylmethylxanthine, dibutyryl cyclic AMP, milrinone and piroximone; unchanged by increased [Ca2+]o and prolonged by caffeine, suggesting that the rates of Ca2+ release and uptake by the sarcoplasmic reticulum were respectively increased, unchanged or decreased by these groups of drugs. Relative to changes in [Ca2+]o, the ratio of the peak of the aequorin light signal to the peak of the tension response was increased by isoprenaline, milrinone and piroximone, and decreased by caffeine, indicating that the Ca2+ sensitivity of the myofilaments was respectively decreased, and increased by these drugs. The effects of milrinone and piroximone on the amplitude and time course of the aequorin light signal, as they relate to changes in uptake and release of calcium from the sarcoplasmic reticulum and to changes in the sensitivity of the myofilaments to Ca2+, are consistent with the findings that positive inotropic doses of these agents act by increasing intracellular concentrations of cyclic AMP. Higher doses of milrinone and piroximone produced negative inotropic effects that were characterized by diminution of developed tension but no change or an increase in the amplitude of the aequorin light signal, suggesting a decrease in the sensitivity of the contractile elements to Ca2+. Toxic doses of milrinone, piroximone and isoprenaline were associated with development of a Ca2+-overload state characterized by the presence of after glimmers, after-contractions and dysrhythmias, and by decreased amplitude of both the aequorin light signal and tension response. The negative inotropic and toxic effects of milrinone and piroximone can be explained only in part by increased intracellular concentrations of cyclic AMP; we suggest that these drugs may have other cardiac actions. PMID- 2992657 TI - The effect of nedocromil sodium and sodium cromoglycate on antigen-induced bronchoconstriction in the Ascaris-sensitive monkey. AB - Nedocromil sodium inhibited the bronchoconstriction caused by antigen challenge in Ascaris-sensitive monkeys and in addition it prevented the release of histamine from mast cells lavaged from sensitive monkeys. Sodium cromoglycate was relatively inactive in both these systems. It is suggested that nedocromil sodium can stabilize both mucosal and connective tissue mast cells and may represent a new type of drug. PMID- 2992659 TI - Subcellular localization of [3H]-nitrendipine binding sites in guinea-pig ileal smooth muscle. AB - The binding of [3H]-nitrendipine was studied in microsomal fractions isolated from guinea-pig ileal smooth muscle. Only one class of specific binding sites was detected, with a KD of 0.4 nM. For various dihydropyridine derivatives, including the stereoisomers of nimodipine and the 'Ca agonist' Bay K 8644, the potency for inhibition of [3H]-nitrendipine binding correlated well with the reported pharmacological potency in smooth muscle preparations. To establish the subcellular localization of [3H]-nitrendipine binding sites, untreated and digitonin-treated microsomal fractions were subfractionated by isopycnic density gradient centrifugation. The density distribution of [3H]-nitrendipine binding was markedly shifted by digitonin towards higher densities, as were the distributions of 5'-nucleotidase and [3H]-ouabain binding, whereas the distributions of NADPH:cytochrome c reductase and NADH:cytochrome c reductase were hardly modified by digitonin. It is concluded that most, if not all, [3H] nitrendipine binding sites in guinea-pig ileal smooth muscle are present in the plasma membrane, in agreement with the postulated mode of action of dihydropyridines as inhibitors of plasmalemmal Ca channels. PMID- 2992658 TI - Erythrosin B inhibits high affinity ouabain binding in guinea-pig heart Na+-K+ ATPase without influence on cardiac glycoside induced contractility. AB - Binding of [3H]-ouabain to guinea-pig heart membranes enriched in Na+-K+-ATPase revealed two different cardiac glycoside binding sites. High affinity binding was obtained at a KD = 2.2 X 10(-7) mol 1(-1) (Bmax = 16.8 pmol ouabain mg-1 protein) whereas low affinity ouabain binding occurred at a KD much greater than 10(-6) mol 1(-1). To discover whether the two ouabain binding sites are functional in guinea-pig heart muscle, erythrosin B, an inhibitor of the high affinity ouabain binding in rat brain tissue, was tested in guinea-pig isolated heart muscle preparations. Erythrosin B proved to be a potent inhibitor of the Mg2+ (Na+) dependent-, as well as Na+-K+-activated ATPase (ID50 = 9 X 10(-6) mol 1(-1). Contractility of guinea-pig isolated papillary muscles, however, was not influenced by erythrosin B in concentrations up to 1 X 10(-5) mol 1(-1). Only very high concentrations (4 X 10(-4) mol 1(-1) resulted in a slightly negative inotropic effect (about 20%). Erythrosin B dose-dependently inhibited [3H] ouabain binding to the Na+-K+-ATPase (KD = - 3.6 X 10(-6) mol 1(-1). In a concentration of 1 X 10(-5) mol 1(-1) the dye abolish high affinity [3H]-ouabain binding without affecting the low affinity binding sites. In contrast, in guinea pig isolated atria, no functional antagonism between erythrosin B (5 X 10(-5) mol 1(-1) and ouabain was observed. 5 As there is a coincidence between the high affinity binding (KD = 2.2 x 10-7moll ') and the concentration for half maximum inotropic effects of ouabain (EDIo = 1.6 x 10-7 mol I), the lack of effect oferythrosin B on ouabain-induced inotropy may be caused by an inaccessibility of the dye to the (internal) ATP-site of the Na+-K+-ATPase. PMID- 2992660 TI - Autoradiographic localization of beta-adrenoceptor subtypes in guinea-pig kidney. AB - The distribution of beta-adrenoceptor subtypes in slide-mounted sections of guinea-pig kidney has been examined by the technique of in vitro labelling combined with autoradiography. Binding of (-)-[125I]-cyanopindolol (Cyp) to kidney sections equilibrated and dissociated slowly, was saturable and stereoselective with respect to the isomers of propranolol and pindolol. These characteristics were appropriate for binding to beta-adrenoceptors. Delineation of beta-adrenoceptor subtypes was achieved by use of betaxolol (beta 1 adrenoceptors) and ICI 118,551 (beta 2-adrenoceptors) and computer assisted curve fitting techniques. Both beta 1- and beta 2-adrenoceptors were present in the proportions 1:2. 3H-Ultrofilm images of (-)-[125I]-Cyp binding to guinea-pig kidney sections showed localized patches of binding in the cortex and concentrated binding in the outer stripe of the medulla. Cortical receptors were of the beta 1 subtype and those associated with the outer stripe of the medulla were of the beta 2-adrenoceptor subtype. beta 1-Adrenoceptors were concentrated over glomeruli and beta 2-adrenoceptors over the straight portion of the proximal tubule. PMID- 2992661 TI - Different responsiveness of prostaglandin D2-sensitive systems to prostaglandin D2 and its analogues. AB - Prostaglandin D2 (PGD2) and six PGD2 analogues were used to classify responsiveness of several PGD2-sensitive systems. The analogues used were 9 beta PGD2, 5-(6-carboxyhexyl)-1-(3-cyclohexyl-3-hydroxypropyl)hydantoin (BW245C), 17 phenyl-18,19,20-trinor-PGD2 (17-phenyl-PGD2), PGD2 amide, PGD2 N-monomethylamide and 11-keto-15 alpha-hydroxy-delta 5,9,12-prostenoic acid (9-deoxy-delta 9,12 PGD2). The PGD2-sensitive systems examined were human platelets, rat peritoneal mast cells, rabbit transverse stomach strip, guinea-pig tracheal ring chain and helical strip of the dog cerebral artery. PGD2, 9 beta-PGD2 and BW245C inhibited the aggregation of human platelets, increased adenosine 3'5'-cyclic monophosphate (cyclic AMP) in rat mast cells and relaxed the rabbit stomach strip. The rank order of potency was BW245C greater than PGD2 greater than 9 beta-PGD2. PGD2 amide and PGD2 N-monomethylamide were inactive in the former two systems but elicited relaxant activity on the rabbit stomach strip. 17-Phenyl-PGD2 was virtually inactive in the above three systems. PGD2 and 17-phenyl-PGD2 contracted the guinea-pig tracheal ring chain and the helical strip of dog cerebral arteries with almost equal potency. 9 beta-PGD2 and BW245C antagonized competitively the contractile action of PGD2. PGD2 amide and PGD2 N-monomethylamide showed weak agonistic actions in the tracheal preparation. 9-Deoxy-delta 9,12-PGD2, showing stronger growth inhibition than PGD2 on cultured tumour cells, was inactive in human platelets, rat mast cells and guinea-pig trachea, and elicited contractile response in the rabbit stomach strip. These results indicate the presence of three groups of PGD2-sensitive systems that respond differently to PGD2 and its analogues. PMID- 2992662 TI - Pharmacological modulation of responses of guinea-pig airways contracted with arachidonic acid. AB - Arachidonic acid (AA) was used to induce contractions of guinea-pig tracheal and lung parenchymal preparations in the presence of indomethacin. Prior addition of FPL55712, nordihydroguaiaretic acid (NDGA), piriprost, benoxaprofen or nafazatrom, in order of potency, inhibited AA-induced contractions of trachea. Higher concentrations (2 - 3 fold) were necessary to inhibit contractions of parenchyma. FPL55712 and piriprost appeared to act as pharmacological antagonists of leukotrienes because they rapidly reduced the tone of the airways established by AA. Administration of exogenous AA to indomethacin-treated trachea appears to be a good model to examine leukotriene receptor antagonists and inhibitors of the lipoxygenase pathway. PMID- 2992663 TI - Selective effects of thiol reagents on the binding sites for imipramine and neurotransmitter amines in the rat brain. AB - The action of the antithyroid drugs methimazole (MMI) and propylthiouracil (PTU) on the binding of [3H]-imipramine, [3H]-5-hydroxytryptamine [3H]-5-HT) (to 5-HT1 receptors) and [3H]-spiperone (to 5-HT2-, D2-receptors) of rat brain membranes has been examined. The synaptosomal uptake of [3H]-5-HT was also studied. Micromolar concentrations of the disulphide bond reducing agents MMI, PTU, dithiothreitol (DTT) and mercaptoethanol increased both the binding of [3H] imipramine and the uptake of [3H]-5-HT. In contrast, they decreased the number of 5-HT1-receptors, and did not affect 5-HT2-and D2-sites. Reaction with membrane bound sulphydryl (SH) groups by micromolar concentrations of N-ethylmaleimide (NEM), hydroxymercuribenzoic acid (PCMB), or Ellman's reagent (DTNB) decreased the binding of [3H]-imipramine, the number of 5-HT1-receptors, and the uptake of [3H]-5-HT. Millimolar concentrations of NEM were necessary in order to decrease partially 5-HT2- and D2-receptors. The effects of NEM on imipramine recognition sites and on the uptake of 5-HT could be prevented by DTT; protection was not obtained in other receptor systems. Three groups of receptors have been, thus, postulated, based upon their different sensitivity towards alterations in membrane [disulphide bridges in equilibrium SH] equilibrium: Group I, including imipramine recognition sites and the uptake system for 5-HT; Group II, including 5-HT1-receptors; Group III, including 5-HT2-and D2-receptors. PMID- 2992664 TI - Potentiation by TRH of the effect of imipramine on the forced-swimming test. AB - Discovery of the potentiation of thyrotropin releasing hormone (TRH)-induced hyperthermia in mice by antidepressants which activate alpha-adrenergic systems instigated investigation of other relations between TRH and antidepressants. For this study the forced-swimming test using mice was chosen since this test is more sensitive for selection of antidepressants which modify catecholaminergic systems than for those affecting 5-hydroxytryptaminergic systems. The effects of imipramine were potentiated by TRH. The involvement of alpha-adrenergic systems was then investigated in this effect since it is already known that these systems are directly implicated in the potentiation of TRH-induced hyperthermia by some antidepressants. Then the involvement of opiate systems was investigated since endogenous opiates are implicated in the action of some antidepressants, and some interactions between TRH and opiate systems are known to exist. TRH made effective a completely inactive dose of imipramine as small as 2 mg kg-1 (i.p.) or 1 microgram per mouse (i.c.v.). Pretreatment by both alpha 1- and alpha 2 adrenoceptor antagonists (phenoxybenzamine, 8 mg kg-1 i.p.; phentolamine, 4 mg kg 1 i.p.) or by a alpha 1-adrenoceptor antagonist (prazosin, 2 mg kg-1 i.p.) did not prevent this potentiation. In contrast the alpha 2-adrenoceptor antagonist (Yohimbine, 2 mg kg-1 i.p.) blocked the TRH effect. The imipramine potentiation by TRH was blocked by pretreatment with an opiate antagonist (naloxone, 1 mg kg-1 i.p.) and the potentiation was decreased in morphine-tolerant mice. 5 These data indicate that potentiation of the effects of imipramine on the forced-swimming test does not seem to be associated with an increase of effective levels of noradrenaline in the synaptic clefts and suggest an interaction between TRH and the opiate systems. PMID- 2992665 TI - Insensitivity of calcium-dependent endothelial stimulation in rat isolated aorta to the calcium entry blocker, flunarizine. AB - In rat aortic segments complete with endothelium, acetylcholine (1 microM) relaxed noradrenaline, phenylephrine and prostaglandin F2 alpha (PGF2 alpha) induced contractions of various magnitudes. Maximal 1 microM phenylephrine induced contractions were relaxed to a greater extent than were maximal contractions induced by the other two agonists. Contractions elicited by various concentrations of phenylephrine and PGF2 alpha in the presence of a maximal effective concentration of the calcium entry blocker flunarizine (3 microM) were relaxed by acetylcholine to about the same residual tension as were contractions elicited in the absence of flunarizine. Acetylcholine (1 microM) and phenylephrine (1 microM) increased tissue levels of guanosine cyclic 3'5' monophosphate (cyclic GMP) by about 37 fold and 2 fold respectively. Preincubation of tissues in the absence of calcium abolished these agonist induced increases in cyclic GMP levels, but preincubation with flunarizine had no significant effect on the increase in cyclic GMP level induced by the agonists. Pretreatment with flunarizine had no significant effect on the basal tissue level of cyclic GMP, but pretreatment in calcium-free solution reduced the basal tissue level of the cyclic nucleotide by about half. It is concluded that in rat aorta, endothelium-dependent acetylcholine-induced relaxation and endothelium-dependent acetylcholine and phenylephrine-induced increases in tissue levels of cyclic GMP, are dependent on extracellular calcium, but are not antagonized by flunarizine. This may indicate that if calcium channels of endothelial cells are activated by these agonists, their characteristics are not identical with those of the calcium channels of the smooth muscle cells. PMID- 2992666 TI - Histamine-induced inositol phospholipid breakdown in the longitudinal smooth muscle of guinea-pig ileum. AB - The characteristics of histamine-stimulated inositol phospholipid breakdown in slices of guinea-pig ileal smooth muscle and cerebellum have been investigated. In cerebellar slices the inhibition of the inositol phospholipid response to histamine by mepyramine was consistent with competitive antagonism of histamine H1-receptors. In slices of the longitudinal smooth muscle of guinea-pig ileum, mepyramine produced only a weak inhibition of the response to histamine, at concentrations up to 1 microM. This was in striking contrast to the potent competitive antagonism of the H1-mediated contractile responses obtained with mepyramine in this tissue. The H1-receptor antagonists (+)-chlorpheniramine and promethazine similarly had no effect on the EC50 value for histamine in guinea pig ileum, while promethazine competitively antagonized the muscarinic receptor mediated inositol phospholipid response in this tissue (Ka 3.6 X 10(7)M-1). Cimetidine, on its own, did not significantly inhibit the inositol phosphate accumulation elicited by histamine in ileum. In the presence of 0.2 microM mepyramine, cimetidine (0.1 mM) produced a small parallel shift of the histamine concentration-response curve (Ka 3 X 10(4) M-1). This inhibition, however, was not consistent with antagonism of an H2-receptor-mediated response. The effect of a range of histamine analogues on inositol phospholipid breakdown was determined. Dose-response curves were constructed and characterized in terms of the EC50, slope and maximal response attainable relative to histamine. The H1-agonists, N alpha,N alpha-dimethylhistamine, N alpha-methylhistamine, 2-pyridylethylamine and 2-thiazolyethylamine produced the largest accumulations of [3H]-inositol-1 phosphate. A very weak response was produced by the H2-selective agonist impromidine, while dimaprit (also H2-selective) was without significant effect. Mepyramine appeared to antagonize competitively the response to the H1-selective agonist 2-pyridylethylamine. This was in contrast to the data obtained with other H1-agonists, where mepyramine produced only a small dextral shift of the agonist curves at low agonist concentrations and an increase in the Hill coefficient. This was particularly striking in the case of 2-methylhistamine. The results suggest that an H1-receptor component in guinea-pig ileum, may coexist with a larger inositol phospholipid response to histamine which is independent of the activation of H1- or H2-receptors. PMID- 2992667 TI - Carbachol and dibutyryl cyclic GMP on the vulnerability to ventricular fibrillation in rat isolated hearts. AB - The hypothesis that elevation of intracellular guanosine 3':5' cyclic monophosphate (cyclic GMP) concentrations may increase electrical stability of the myocardium was examined by determination of ventricular fibrillation thresholds (VFT) on isolated perfused hearts of the rat. Hearts were paced to circumvent any complicating effects of bradycardia. Using this system, carbachol produced a concentration-related reduction in VFT. The reduction in VFT produced by carbachol was not significantly modified by a high concentration of atenolol (10(-5)M), indicating that the increased vulnerability to ventricular fibrillation was not an indirect consequence of catecholamine release from intramyocardial stores. Atropine (10(-6)M) blocked the carbachol-induced reduction in VFT. At the concentrations of carbachol used to reduce VFT, myocardial cyclic GMP concentrations were also elevated. The dibutyryl analogue of cyclic GMP (10(-4)M) mimicked the effect of carbachol in reducing VFT. Carbachol potentiated the adrenaline (3 X 10(-7)M)-induced reduction in VFT. PMID- 2992668 TI - Effects of chronic administration of amitriptyline or mianserin on rat cardiac and central adrenoceptors. AB - Rats were administered either amitriptyline (20 mg kg-1, i.p.) or mianserin (10 mg kg-1, i.p.) for 21 consecutive days and the alpha- and beta-adrenoceptor characteristics of cardiac ventricles, cerebral cortex and hippocampus examined by ligand-binding procedure. Chronic administration of amitriptyline significantly reduced the maximum density of beta-receptors in the cerebral cortex without significantly altering cardiac alpha- or beta-receptors; mianserin treatment had no significant effect on any of the receptors studied. PMID- 2992669 TI - Pre-incubation of guinea-pig myenteric plexus with beta-funaltrexamine: discrepancy between binding assays and bioassays. AB - The acute effects of beta-funaltrexamine and the effects of pre-incubation with this compound were examined in five in vitro assay tissues and in selective binding assays in homogenates of guinea-pig brain and myenteric plexus. In competitive displacement assays with selective ligands, beta-funaltrexamine had highest affinity for the mu-binding site in the myenteric plexus and brain of guinea-pig. Its affinity for the kappa-site was about 15% of that for the mu site. Pre-incubation of the assay tissues with beta-funaltrexamine caused an increase in the IC50 values of mu- and delta-receptor agonists but not of kappa agonists. Although in bioassays on the myenteric plexus-longitudinal muscle preparation of the guinea-pig, the IC50 value of the mu-receptor ligand [D-Ala2, MePhe4, Gly-ol5] enkephalin was increased up to 124 fold, its binding at the mu site in homogenates of the preparation was not affected by this treatment. These findings indicate that the effects of pre-incubation with beta-funaltrexamine on agonist potency of the mu-receptor ligand are due to an interference with the coupling mechanism between the mu-binding site and the effector system. PMID- 2992670 TI - Prolongation of inhibitory postsynaptic currents by pentobarbitone, halothane and ketamine in CA1 pyramidal cells in rat hippocampus. AB - Spontaneous inhibitory postsynaptic currents (i.p.s.cs) were recorded in voltage clamped CA1 neurones in rat hippocampal slices. The exponential decay of i.p.s.cs was prolonged by concentrations of sodium pentobarbitone as low as 50 microM. With concentrations up to 100 microM, there was no change in the amplitude or rise time of the currents but current amplitude was depressed at 200 microM. The prolongation of currents increased with drug concentration within the range tested (50 to 200 microM). Halothane, at concentrations from 1 to 5%, also increased the time constant of decay of i.p.s.cs. The effect increased with concentration and was fully reversible. Ketamine, at a concentration of 0.5 mM, increased the time constant of decay of i.p.s.cs by 50 to 80% and the effect was reversible. Ethanol (10-200 mM), nitrous oxide (75-80%), and caffeine (10 microM 5 mM) had no detectable effect on the i.p.s.cs. It is suggested that pentobarbitone, halothane and ketamine increase the time constant of decay of the i.p.s.cs by stabilizing the open state of channels activated by gamma aminobutyric acid. PMID- 2992672 TI - Biochemical peculiarities of trypanosomes, African and South American. PMID- 2992671 TI - Central delta-opioid receptor interactions and the inhibition of reflex urinary bladder contractions in the rat. AB - The in vivo effects of a number of opioid agonists and antagonists were studied on the spontaneous reflex contractions of the urinary bladder recorded isometrically in the rat anesthetized with urethane. All substances were administered into the central nervous system by the intracereboventricular (i.c.v.) or spinal intrathecal (i.t.) route. The conformationally restricted enkephalin analogues [2-D-penicillamine, 5-L-cysteine] enkephalin (DPLCE), [2-D penicillamine, 5-L-penicillamine] enkephalin (DPLPE) and [2-D-penicillamine, 5-D penicillamine] enkephalin (DPDPE) produced dose-related inhibition of reflex bladder contractions when administered by the i.c.v. or i.t. route. Both the novel delta-opioid receptor antagonist ICI 154,129 (200-600 micrograms) [N,N bisallyl-Tyr-Gly-Gly-Psi-(CH2S)-Phe-Leu-OH) and ICI 174,864 (1-3 micrograms) [N,N dially-Tyr-Aib-Aib-Phe-Leu-OH: Aib = alpha-aminoisobutyric acid] attenuated or abolished the effects of DPLCE, DPLPE and DPDPE when administered by the i.c.v. or i.t. route. The antagonism observed was selective since the equipotent inhibition produced by the mu-opioid receptor agonist [D-Ala2, Me-Phe4, Gly(ol)5] enkephalin (DAGO) was unaffected. Overall, ICI 154,129 was considerably weaker than ICI 174,864 and both antagonists inhibited bladder activity at doses higher than those required to demonstrate delta-receptor antagonism. Further studies of the agonistic effect of ICI 174,864 showed that it was insensitive to low doses of naloxone (2 micrograms, i.c.v. or i.t.) but could be abolished by higher (10 15 micrograms) doses of naloxone. These observations suggested that the agonistic effect of ICI 174,864 was not mediated by mu-opioid receptor. beta-Endorphin (0.2 1.0 micrograms, i.c.v.) inhibited bladder contractions but following recovery from this effect, appeared to prevent the expression of delta-receptor antagonism by ICI 174,864. In addition a previously subthreshold dose of ICI 174,864 now exhibited marked agonistic activity. The inhibitory effect of a submaximal dose of DPDPE was also potentiated by beta-endorphin under these circumstances. These observations suggest that supra-spinal and spinal delta-opioid receptors are involved in the opioid-mediated inhibition of reflex bladder contractions in the rat. Moreover beta-endorphin may be important in regulating central delta-opioid receptors. PMID- 2992673 TI - Hypothalamo-pituitary responses to trauma. PMID- 2992674 TI - Fluid and electrolyte disturbances after trauma: the role of adrenocortical and pituitary hormones. PMID- 2992675 TI - Impaired phagocytic activity of neutrophils in patients receiving haemodialysis: the critical role of iron overload. AB - The metabolic burst (as measured by the spontaneous and stimulated nitroblue tetrazolium tests), the phagocytosis of heat inactivated bakers' yeast and of Staphylococcus aureus, the killing of Staph aureus, and the myeloperoxidase activity of polymorphonuclear neutrophils were studied in 11 patients receiving maintenance haemodialysis. Of these patients, six were polytransfused and had high serum ferritin concentrations (mean 5940 (SD 2925) micrograms/l; group 1), and five had normal serum ferritin values (mean 171 (116) micrograms/l; group 2). Patients in group 1 had a history of more infectious episodes (0.167 v 0.025 per patient per month) and significantly more genitourinary infections (p = 0.015) than those in group 2. Phagocytosis and myeloperoxidase activity were severely reduced in group 1 but normal in group 2. Percentages of neutrophils ingesting one or more particles together with the index of phagocytosis in patients' serum were inversely correlated with serum ferritin concentrations. Four patients in group 1 were treated with desferrioxamine, and after six to 18 weeks of treatment phagocytosis and myeloperoxidase activity had returned to normal in three of them. These data suggest that in patients receiving haemodialysis iron overload due to multiple transfusions plays an important part in the mechanisms underlying the susceptibility to bacterial infections, mediated at least partially through impaired neutrophil function. PMID- 2992677 TI - Immunohistochemical detection of the gene product of Rous sarcoma virus in human brain tumors. AB - We have studied 108 cases of brain tumors by immunohistochemical staining utilizing an antiserum against the src gene product and GFA protein in order to elucidate the relationship between the brain tumors and the transforming genes. While the normal human brain tissues were not positive with the antiserum against the src gene product, several astrocytomas were clearly positive and other brain tumors were negative with the same. Src-gene product related peptide was detected in some of the human astrocytomas. PMID- 2992676 TI - HLA-DRw6 as a risk factor for active cytomegalovirus but not for herpes simplex virus infection after renal allograft transplantation. AB - To study genetically determined susceptibility to cytomegalovirus and herpes simplex virus infections in patients given renal transplants a prospective study was performed of 68 consecutive patients receiving their first cadaveric kidney allograft. The recipients positive for HLA-DRw6 showed a significantly increased incidence of active cytomegalovirus infection as early as the 10th week after transplantation (p less than 0.05). No relation with other human leucocyte antigens was found, nor did a correlation exist between HLA typing and the incidence of herpes simplex virus infections. Furthermore, recipients positive for HLA-DRw6 with secondary cytomegalovirus infections excreted infectious virus more often (p less than 0.01) and showed more clinical symptoms (p less than 0.01) than a comparable group of recipients negative for HLA-DRw6. These observations may have practical implications for the treatment of patients who have had renal transplant operations. PMID- 2992678 TI - Studies on spinal opiate receptor pharmacology. III. Analgetic effects of enkephalin dimers as measured by cutaneous-thermal and visceral-chemical evoked responses. AB - D-Ala2-D-Leu5-enkephalin (DADL) along with dimers formed from tetra(DTE: des-Leu5 enkephalin)- or pentapeptide (DPE: Leu5-enkephalin) coupled by methylene bridges of various lengths (n) have been shown in in vitro systems to possess varying degrees of mu/delta-receptor selectivity. In the present studies we have systematically compared the intrathecal effect of these agents and morphine on the cutaneous stimuli (hot plate (HP) and tail flick (TF) and visceral-chemical (writhing) tests in the rat. The following observations were made. (1) Dimers with high delta-receptor selectivity were active in the TF(DPE2 greater than or equal to DADL greater than or equal to DTE2 greater than or equal to morphine greater than DPE12 much greater than DTE12 = 0) and HP(DPE2 greater than or equal to DTE2 greater than or equal to DADL greater than or equal to morphine greater than DPE12 greater than or equal to DTE12 greater than 0. To examine cross tolerance, the intrathecal ED50 for morphine, DPE and DADL were determined in rats rendered tolerant by subcutaneous morphine pellets. The TF ED50 tolerant/TF ED50 naive was 18.4, 5.4 and 1.3, respectively. The ratio of activity on the HP was 14.0, 4.7 and 2.2 (2) On the visceral-chemical test, only morphine was active. The dimers or DADL in doses which totally blocked the TF or HP are at higher doses which were just below those producing motor dysfunction had no effect on the writhing response. (3) At high intrathecal doses (40 X TF ED50), morphine produced a motor rigidity which blocked the placing and stepping reflex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992679 TI - The effects of ethanol on the efflux and release of norepinephrine and 5 hydroxytryptamine from slices of rat hypothalamus. AB - Studies were done to determine the effects of ethanol on release of norepinephrine, dopamine and 5-hydroxytryptamine from nerve terminals in the central nervous system. Superfused slices of rat hypothalamus were used in these studies and endogenous amines in the superfusate were quantitated using HPLC with electrochemical detection. In these experiments 'release' of transmitters was studied in the presence of amitriptyline to block neuronal uptake of amines, whereas 'efflux' was measured in its absence. A highly intoxicating concentration of ethanol (69.6 mM, 320 mg%) increased the K+-evoked release of norepinephrine, dopamine and serotonin without affecting basal release. Since this concentration of ethanol increased the basal efflux but not the basal release of 5 hydroxytryptamine, it appeared that neuronal uptake of 5-hydroxytryptamine under basal conditions may also be inhibited by intoxicating levels of ethanol. PMID- 2992680 TI - Is taurine a neurotransmitter in rabbit retina? AB - Rabbit retina was used as a model to study the possible role of taurine in the retina. The taurine-synthesizing enzyme, cysteine sulfinic acid decarboxylase (CSAD), is localized immunohistochemically using specific antibodies against CSAD. The CSAD-immunoreactivity appears to be most prominent in the inner nuclear layer (INL) and ganglion cell layer (GCL). The inner plexiform layer (IPL), the outer nuclear layer and outer plexiform layer are sporadically stained. The CSAD positive neurons include some amacrine cells and probably the bipolar cells in the INL and some large and small ganglion cells in the GCL. Autoradiographic studies reveal that the uptake of [3H]taurine is most prominent in the INL. The IPL and GCL, as well as the Muller cells, also show a moderate degree of [3H]taurine accumulation. In conclusion, we have demonstrated the presence of the taurine-synthesizing enzyme and uptake systems in rabbit retina. Based on the above evidence, we propose that taurine may be used by some neurons, presumably amacrine cells, as a transmitter in the rabbit retina. PMID- 2992681 TI - Hyperexcitability following moderate hypoxia in hippocampal tissue slices. AB - The effects of moderate hypoxia on CA1 pyramidal cells were studied in submerged rat hippocampal tissue slices. Hyperexcitability developed during re-oxygenation following 30-45 min of hypoxia at 29 degrees C. During hypoxia a sustained increase in extracellular potassium was observed. The post-hypoxic hyperexcitability of CA1 neurons may be the result of their inability to fully recover intracellular potassium lost during oxygen deprivation. PMID- 2992682 TI - Multi-dimensional analyses of behavior in mice treated with U-50,488H, a purported kappa (non-mu) opioid agonist. AB - The effects of U-50, 488H (trans-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl) cyclohexyl ]-benzeneacetamide, methane sulfonate, hydrate), a purported selective kappa (non-mu) opioid agonist on spontaneous locomotor activity were investigated using a multi-dimensional behavioral analyzer (Animex II). U-50,488H (1 mg/kg) failed to affect behavior in mice, however, 3 mg/kg significantly reduced rearing and grooming. In addition, 10 mg/kg markedly reduced linear locomotion, rearing and grooming. The behavioral depression induced by U-50,488H (10 mg/kg) was reversible by the opioid antagonist Mr2266 (10 mg/kg). These results suggest that the selective activation of the kappa (non-mu) opioid receptor by U-50,488H decreases linear locomotion, rearing and grooming in mice. PMID- 2992683 TI - Ventromedial thalamic lesions and seizure susceptibility. AB - Recent data indicate that the substantia nigra is an important site in a circuitry involved in the modification of various experimental seizures with neocortical and limbic involvement. Since there are no direct nigral projections to either area, we assumed that the nigral effects on seizures are relayed by other sites such as the thalamus. To evaluate this hypothesis we produced bilateral high-radiofrequency thermocoagulative lesions of the ventromedial (VM) thalamic nuclei which receive the nigral efferents in the rat. We determined the susceptibility of lesioned and control adult rats to the development of flurothyl seizures 2 and 4 weeks later. The latency to the onset of a generalized seizure was considered as the convulsive threshold. There were no differences in the mean latencies between the groups. The results suggest that bilateral destruction of the VM thalamic nuclei does not modify the susceptibility to the development of flurothyl seizures in the rat. PMID- 2992684 TI - Neuron generator potentials evoked by intracellular injection of cyclic nucleotides and mechanical distension. AB - Depolarization of neuron membrane was shown to occur during cAMP injection by means of both iontophoresis and pressure. Distension of the neuron by means of blowing with large volumes of solution without cAMP can produce reversible responses. The time course can be made similar to that of the cAMP effect and the responses can be repeated many times in the same neuron. The effect of cAMP can be differentiated from the mechanical response by injecting small volumes of concentrated solutions. The delay in this depolarization response to cAMP is only 0.1-0.3 s. The calculated time of diffusion from the electrode tip to the membrane is about 10 times greater. The similarity of cAMP and mechanical responses and the short delay of the cAMP effect suggests that a cAMP effect may be mediated by mechanical signals in the cytoskeleton. PMID- 2992685 TI - Presynaptic actions of cholinergic agents upon the hair cell-afferent fiber synapse in the vestibular labyrinth of the frog. AB - Spontaneous activity of semicircular canal afferents in the isolated labyrinth of the frog is altered by bath application of cholinergic agonists. Muscarinic agonists can produce an increase in action potential frequency of individual afferents. This increase develops slowly and is prolonged in the time course of its action. Nicotinic agonists can either increase (most cases) or decrease afferent activity. These effects occur rapidly and decay during the period of activation, suggesting desensitization. Muscarinic effects are blocked by prior administration of atropine and nicotinic effects (both increases and decreases in action potential frequency) by curare. Intracellular recordings reveal that the nicotinic effects on afferent action potential frequency are the result of alterations in the frequency of spontaneous synaptic potentials, indicating a presynaptic site of action on the hair cells for these compounds. This conclusion is supported by the fact that in the presence of high Mg2+/low Ca2+, which blocks hair cell release of transmitter, cholinergic agonists do not affect the resting membrane potential of the vestibular afferent. Electrical stimulation of the VIIIth cranial nerve can result in either an increase or a decrease in spontaneous synaptic potential and action potential frequency of an afferent. These effects are blocked by prior administration of curare or of nicotinic agonists. Repetitive or continuous stimulation of the VIIIth nerve results in a reversible reduction of the evoked response, suggesting desensitization. Transection of the VIIIth cranial nerve two weeks prior to recording eliminates these actions of electrical stimulation, but not the responses to cholinergic agonists, indicating that the effects of electrical stimulation are mediated by centrally arising efferents. These findings confirm that acetylcholine is probably the transmitter released from centrally arising vestibular efferents, and, in addition, demonstrate that efferent-mediated effects are predominantly expressed through nicotinic receptors. Studies comparing the effects of isolation of the semicircular canal alone versus the intact labyrinth suggest that the method of isolation may be an important factor in determining whether efferent activity results in a predominant increase or decrease in afferent activity. PMID- 2992686 TI - Cytochrome reduction coincides with electrical activity in perfused bullfrog brain. AB - The redox states of cytochromes a(a3), b and c + c1 were continuously measured on the surface of the perfused bullfrog brain with the aid of a scanning spectrophotometer. To correlate with changes of the redox state, electrical activity was simultaneously recorded in the vicinity of the tip of the light guide of the spectrophotometer. In most cases, parallel reduction of the cytochromes was recorded in concordance with burst activity when the third or the fourth dorsal root of the spinal cord was electrically stimulated for 2 min. Recurrent cytochrome reduction and burst activity were produced by stimulation for a longer period, 5 or 10 min. Both the cytochrome reduction and burst activity were abolished when 1 microM tetrodotoxin was added to the perfusing solution. These results suggest that the cytochrome reduction coinciding with electrical activity in the brain may correlate with the Na+-influx followed by the activation of Na+,K+-adenosine triphosphatase in the neurons. PMID- 2992687 TI - In vivo modulation of norepinephrine-induced cerebral oxygenation states by hypoxia and hyperoxia. AB - The effect of intravenous norepinephrine (NE) administration on three O2 dependent parameters of cerebral oxygenation was studied in the parietal cortex of skull intact anesthetized rats. Reflectance spectrophotometry was used to measure in vivo changes in cortical hemoglobin saturation (Hb/HbO2), blood volume (BV), and cytochrome c oxidase (cyt. a,a3) oxidation-reduction state. The influence of arterial pressure of oxygen (paO2) on norepinephrine-induced changes in cortical microcirculatory O2 delivery and cyt. a,a3 redox state was tested under conditions of normoxia, hypoxia, and hyperoxia. Norepinephrine produced cyt. a,a3 redox changes which were independent of compensatory alterations in cortical blood volume and changes in systemic blood pressure at the tested physiological extremes. During normoxia, NE caused dose-dependent systemic pressure-related increases in the oxidation level of cyt. a,a3. Conversely, in hypoxia NE caused a reduction. Microcirculatory and cyt. a,a3 redox responses to low doses of NE during hyperoxia were similar to those obtained at high doses during normoxia. The kinetic pattern of changes in hemoglobin saturation, cyt. a,a3 redox state, and cortical blood volume during normoxia and hypoxia was consistent with direct alteration in oxygen delivery to the respiratory chain and possible modification of cerebral oxidative metabolism. Blood-brain barrier alterations and vascular smooth muscle resistance changes to NE under tested conditions of oxygenation are postulated to be responsible for the observed results. PMID- 2992688 TI - In vivo voltammetric and behavioural evidence for somatodendritic autoreceptor control of mesolimbic dopamine neurones. AB - Somatodendritic autoreceptor control of mesolimbic dopamine neurones was assessed using in vivo differential pulse voltammetry to monitor dopamine metabolism in the nucleus accumbens following infusion of haloperidol or dopamine into the ipsilateral ventral tegmental area (VTA) of chloral hydrate anaesthetized rats. Haloperidol (2.5 micrograms/0.5 microliters) produced an increase in dihydroxyphenylacetic acid (DOPAC) levels (45%) whilst dopamine produced a transient decrease (38%). In three experiments this decrease was smaller and was followed by a large increase in DOPAC levels (108-208%). However, the results of radiolabelled drug diffusion studies suggested that the secondary rise in DOPAC in the nucleus accumbens may be due to diffusion of dopamine following infusion into the VTA. In a separate set of experiments rats were injected with haloperidol (2.5 micrograms/0.5 microliters) bilaterally into the VTA and their behaviour monitored using doppler shift radar. Haloperidol infusion produced a phase of increased exploratory behaviour lasting approximately 30 min. Both sets of data support previous electrophysiological studies suggesting the presence of functional dopamine somatodendritic autoreceptors in the mesolimbic system. The results further demonstrate the applicability of in vivo voltammetry for study of the effects of drugs on discrete receptor populations. PMID- 2992689 TI - Late evolutionary appearance of 'peripheral-type' binding sites for benzodiazepines. AB - Four classes of non-mammalian vertebrates were examined for the presence of both 'brain-specific' and 'peripheral-type' binding sites for benzodiazepines in the central nervous system. 'Brain-specific' binding sites for benzodiazepines were found in the central nervous systems of all non-mammalian vertebrates studied. However, in contrast to mammals, either very low or undetectable levels of 'peripheral-type' binding sites for benzodiazepines were observed in the central nervous systems of these non-mammalian vertebrates. Furthermore, the density of 'peripheral-type' binding sites for benzodiazepines in non-mammalian vertebrate heart was less than or equal to 2% of that found in mammalian cardiac tissue. These findings suggest a very late evolutionary appearance of 'peripheral-type' binding sites for benzodiazepines, implying that these sites may have (a) highly specialized function(s) in both peripheral tissues and the central nervous system. PMID- 2992690 TI - Inhibition from locus coeruleus of nucleus accumbens neurons activated by hippocampal stimulation. AB - Electrophysiological studies using rats were performed to examine the influence of locus coeruleus (LC) on nucleus accumbens (Acc) neurons. Spike generation by hippocampal stimulation was inhibited by both LC conditioning stimulation and iontophoretic application of noradrenaline, but spikes elicited by stimulation of parafascicular nucleus of thalamus were rarely affected by LC conditioning stimulation or noradrenaline. The LC-induced inhibition was antagonized by iontophoretic sotalol, but not by phentolamine, suggesting that noradrenaline derived from the LC inhibits the Acc neurons receiving input from the hippocampus, probably acting on a beta-adrenergic receptor. PMID- 2992691 TI - Vasopressin-mediated slow EPSPs in a mammalian sympathetic ganglion. AB - Vasopressin is one of numerous neuropeptides contained in sympathetic ganglia, but whose function remains unresolved. In this report, we present electrophysiological evidence that arginine-vasopressin (AVP) is a neurotransmitter in guinea pig inferior mesenteric ganglion (IMG). AVP superfused over the IMG, in vitro, produced in a population of neurons a membrane depolarization accompanied by a resistance increase, both of which were blocked by a specific V1 receptor antagonist. Moreover, slow excitatory postsynaptic potentials (EPSPs) elicited by repetitive nerve stimulation were attenuated in 75% of cells tested in the presence of excess AVP, and occasionally in the presence of the antagonist. Thus, AVP joins substance P as a putative transmitter of slow potentials in the guinea pig IMG. PMID- 2992693 TI - Evidence for impaired GABAergic activity in the substantia nigra of amygdaloid kindled rats. AB - The activity of the GABA-synthesizing enzyme glutamate decarboxylase (GAD) was determined in synaptosomal fractions from 12 brain regions of amygdaloid kindled rats. The only significant difference in regional GAD activities between kindled animals and sham-operated controls was a 40% decrease of GAD activity in the substantia nigra. The data suggest that impaired GABAergic function in the nigra may be involved in the initiation and propagation of amygdaloid-kindled seizures. PMID- 2992694 TI - Newly identified GABAergic neurons in regions of the ventrolateral medulla which regulate blood pressure. AB - Glutamic acid decarboxylase (GAD), the enzyme which synthesizes the inhibitory transmitter gamma-aminobutyric acid (GABA), was localized immunocytochemically within cells and processes distributed throughout the ventrolateral medulla. In caudal regions, GAD-stained cells were adjacent to the 'precerebellar' lateral reticular nucleus and partially overlapped the A1 area of norepinephrine synthesizing neurons. The largest number of labeled neurons filled the rostral ventrolateral medulla (RVL), coinciding with and extending beyond the C1 adrenergic area. GAD-positive cells also occupied the nucleus reticularis parvocellularis, raphe magnus (RM) and lateral wings of RM in the region of the pararaphe. Intrinsic GAD-containing cells in the ventrolateral medulla may tonically inhibit sympathoinhibitory neurons in the caudal ventrolateral medulla (CVL) and sympathoexcitatory neurons in the RVL. PMID- 2992692 TI - A response-specific conditioned aversion to rewarding hypothalamic stimulation in rats. AB - Rats with hypothalamic electrodes were trained to self-stimulate by nose-poking and by licking at a dry spout. Separate groups of animals were then exposed to pairings of either self-stimulation by licking or by nose-poking with the subsequent administration of LiCl. The licking/LiCl pairing group exhibited a marked attenuation of licking maintained by hypothalamic stimulation on the first retention trial (24 h post LiCl). This effect was enhanced by a further licking/LiCl pairing. In a final cross-over trial with this group, nose-poking maintained by stimulation at the same site and intensity was found to be unaffected. With another group of animals nose-poking/LiCl pairing had no effect on self-stimulation, despite two conditioning trials. In a final cross-over trial with this group licking maintained by stimulation of the same site and intensity was also unaffected. These data indicate that the sensory properties of brain stimulation may be dependent on the behavioural context in which the stimulation occurs. PMID- 2992695 TI - Glucocorticoids are modulators of GABAA receptors in brain. AB - Modulation of binding of [3H]muscimol, a GABAA receptor agonist, by natural and synthetic glucocorticoids was investigated in crude synaptosomal membranes and in brain sections of rat. In adrenalectomized (Adx) rats, muscimol binding was reduced by 30-50% in cerebral cortex, cerebellum, thalamus and hippocampus, as compared to sham-operated controls. This decrease was due to reduced binding affinities of GABA receptors for muscimol. In contrast muscimol binding was increased by 38% in the hypothalamus and did not change in the pons-medulla after Adx. Nanomolar concentrations of corticosterone and pregnenolone-sulfate, but not dexamethasone, enhanced muscimol binding in brain regions that were characterized by reduced binding following Adx. This steroid-induced increase in muscimol binding was due to enhanced affinities of GABA receptors. PMID- 2992696 TI - Opiate binding sites in the chick, rabbit and goldfish retina. AB - The characteristics of opiate binding sites in the retina of the chick, rabbit and goldfish have been investigated. In the newly hatched chick retina, 131 fmol/mg of binding sites for [D-Ala2-D-Leu5]-[3H]enkephalin are present; competition studies with the delta selective peptide [D-Thr-Leu5]-enkephalin (DTLET) and the mu selective peptide morphiceptin show that all of the [D-Ala2-D Leu5]-[3H]-enkephalin binding sites are of the delta subtype. Dihydro[3H]morphine binds poorly to the chick retina; 13.2 fmol/mg of this binding is displaceable by morphiceptin and corresponds to mu binding sites. Benzomorphan sites are defined as sites occupied by [3H]diprenorphine which is displaceable by low concentrations of ethylketocyclozacine but not by high concentrations of D-Ala2-D Leu5-enkephalin and morphiceptin. At least 88 fmol/mg of benzomorphan sites are present in the chick retina. [3H]diprenorphine binding to the rabbit and fish retina was measured. The rabbit retina bound 60 fmol/mg, and the fish retina 42 fmol/mg of [3H]diprenorphine. These findings are discussed in the light of the studies on the localization and physiological effects of enkephalin in the retina. PMID- 2992697 TI - Desensitization of beta-receptors on primary astrocyte cultures by norepinephrine but not by tricyclic antidepressants. AB - Primary astrocyte cultures from neonatal rat cerebral hemispheres were treated chronically for up to 3 weeks with the tricyclic antidepressants amitryptyline (AMT) or desipramine (DMI), or acutely with AMT and DMI added at the same time as the agonist, norepinephrine (NE). AMT and DMI were added at concentrations from 10(-9) to 10(-5) M. Both types of treatment did not decrease the increase in cyclic AMP (cAMP) content of these cells in response to a 10 min exposure to 10( 5) M NE. Chronic exposure to the antidepressants also did not affect stimulation of cAMP by isoproterenol (iso) in both rat and mouse primary astrocyte cultures. In contrast to the lack of effect of the tricyclic antidepressants pretreatment of the cultures with 10(-5) M NE resulted in total inhibition of the cAMP response after 2 h, with a 50% decrease occurring in about 45 min. This is similar to the agonist-induced desensitization of the beta-receptor-adenylate cyclase system seen in many other cells. This effect could, in part, be a direct response to increased intracellular cAMP since pretreatment with 0.25 and 1.0 mM N6-2'-O-dibutyryl cAMP (DBcAMP) also resulted in total inhibition of the cAMP response after 4 h. Receptor labelling experiments using [125I]cyanopindolol showed no decreases in apparent binding sites up to 3 h after exposure to 10(-5) M norepinephrine, suggesting that the rapid desensitization of the cAMP response was primarily due to an uncoupling of the receptor from the adenyl cyclase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992699 TI - Membrane potential and input resistance in alpha motoneurons of hindlimb extensors during isolated and clustered episodes of phasic events in REM sleep. AB - The function of motoneurons during the phasic events of REM sleep was investigated in cats. The membrane potential, synaptic activity, input resistance and rheobase were measured in hindlimb extensor motoneurons during periods of concentrated phasic events (Type II) in REM sleep in undrugged, minimally restrained cats. During Type II episodes in REM sleep, motoneurons depolarized 3 mV on the average, reduced in input resistance by 20%, and had distinct increases in spontaneous synaptic activity, as compared with periods of REM sleep between episodes (i.e. Type I). This constellation of properties is explained by increased frequencies of both unitary EPSPs and IPSPs in motoneurons. Myoclonic jerks in Type II episodes do not appear to arise because of a temporary withdrawal of the tonic motoneuron inhibition found in REM sleep. Instead, large increases in excitatory synaptic activity overcome the inhibition, giving rise to motoneuron discharges. PMID- 2992698 TI - Suppression by phenytoin of convulsant-induced afterdischarges at presynaptic nerve terminals. AB - The mechanisms underlying the induction of afterdischarges at presynaptic nerve terminals by convulsant aminopyridines and their suppression by the anticonvulsant drug phenytoin were studied at the frog neuromuscular preparation. Addition of aminopyridine to the perfusing solution induced the appearance of afterdischarges in motor nerve fibres following their primary response to a single nerve stimulus. The afterdischarges seemed to originate at or near the nerve terminals and to propagate both antidromically and orthodromically. The latter resulted in repetitive activation of the neuromuscular synapse. Focal recordings of nerve terminal potentials suggested that aminopyridines may induce afterdischarges by slowing spike repolarization and thereby producing a prolonged depolarization of nerve terminals. Phenytoin suppressed the aminopyridine-induced afterdischarges and the resultant repetitive excitation of the postsynaptic muscle fibres. This effect of phenytoin was associated with a depression of the action potential at the motor nerve terminals but not at their parent axons. These results single the presynaptic nerve terminals as preferential sites for convulsant and anticonvulsant actions. PMID- 2992700 TI - The response of optic tract glia during regeneration of the goldfish visual system. I. Biosynthetic activity within different glial populations after transection of retinal ganglion cell axons. AB - We monitored biosynthetic activity of optic tract glia during regeneration of retinal ganglion cell axons in the goldfish and found that the greatest level of incorporated [3H]thymidine and [3H]leucine occurred in glia by 10-15 days after axotomy. During this period there was a marked increase in the number of oligodendroglia and multipotential glia near the site of injury with no change occurring in the astroglial population. Electron microscopic autoradiography showed that oligodendroglia and multipotential cells incorporated 5-7-fold more thymidine than did cells of intact control preparations. Though all glial cell types incorporated more [3H]leucine during axonal regeneration, oligodendroglia and multipotential cells together accounted for more than 90% of measured radioactivity. In order to characterize glial-stimulating events specific to axonal regeneration, we produced axonal degeneration in the optic tract by removal of the retina. Optic tract glia during axonal degeneration incorporated less amino acid when compared to glia associated with regenerating axons. The degenerating optic tract also had less 2',3'-cyclic nucleotide 3' phosphohydrolase, an enzyme produced by oligodendroglia, than that found in the regenerating visual system. Our results suggest that in response to ganglion cell axotomy oligodendroglia and multipotential glia of the goldfish optic tract proliferate. Moreover, regenerating axons provide one type of stimulant for glial protein biosynthesis. PMID- 2992701 TI - Comparative pharmacological effects on visual cortical neurons in monocularly deprived cats. AB - Monocularly deprived (MD) cats show a loss of responsiveness to visual stimulation of the deprived eye among visual cortical neurons. Several lines of evidence suggest that this effect involves, at least in part, a suppression of deprived eye input, possibly mediated by GABA inhibition. In order to better understand the nature of this suppression we have evaluated the effectiveness of different types of disinhibitory and excitatory agents to reverse the effects of MD. We investigated bicuculline (a GABA antagonist); picrotoxin (a GABA antagonist with a different mechanism of action from bicuculline); strychnine (a glycine antagonist); ammonium ion (a blocker of membrane chloride channels); physostigmine (a cholinesterase inhibitor); and naloxone (an opiate antagonist and also a GABA antagonist). All drugs were given intravenously. Bicuculline restored binocularity to 50% of the visual cortical neurons tested and naloxone to 36%. With both drugs, receptive fields of the normal eye tended to lose specificity. The emergent deprived eye receptive fields were usually similar to those of the normal eye after drug administration. Ammonium ion produced binocular responses in 27% of neurons tested, but receptive fields were grossly abnormal; moreover, ammonium infusion tended to depress neuronal responsiveness. All other drugs tested failed to restore binocularity. These experiments lend further credence to the hypothesis that GABA inhibition contributes to the cortical effects of MD, since only drugs with GABA antagonistic action were effective in restoring neuronal responsiveness to the deprived eye. PMID- 2992702 TI - Spontaneous and evoked release of methionine-enkephalin-like material from the rat spinal cord in vivo. AB - In vivo perfusion of the subarachnoid space with an artificial cerebrospinal fluid (CSF) in paralyzed halothane-anesthetized rats allowed the collection of methionine-enkephalin (Met-Enk)-like material (MELM) released from the spinal cord. Bio-Gel P2 chromatography and high-performance liquid chromatography showed that 65% of this material corresponded to authentic Met-Enk. Under resting conditions, about 1 pg of MELM per minute was regularly released for at least 3 h; for Met-Enk, this value corresponded to a fractional rate constant of 0.002% (i.e. tissue content of the pentapeptide which was released per minute from the whole spinal cord). Perfusion with K+-enriched (40-60 mM) CSF resulted in a marked enhancement (+ 150-200%) of spinal MELM release. Similarly, calibrated pinches of the muzzle and i.p. administration of acetic acid, two strong noxious stimuli in awake animals, induced a significant increase (+ 75-150%) in spinal MELM release. In contrast, pinches applied to the tail did not enhance but instead slightly reduced (-35%) MELM release from the rat spinal cord. These data suggest that mechanisms other than segmental controls could be involved in the activation of spinal enkephalinergic neurons by some nociceptive stimuli. PMID- 2992703 TI - Altered patterns of evoked synaptic activity in cortical pyramidal neurons in feline ganglioside storage disease. AB - Postsynaptic potentials evoked by ventrolateral thalamic stimulation were recorded intracellularly from neurons in the precruciate cortex of GM1 mutants with HRP- or LY-loaded microelectrodes. Ganglioside-laden pyramidal neurons exhibiting somal distention and/or meganeurite formation were found to respond to thalamic stimulation with short duration IPSPs. Evoked EPSPs were recorded from two morphologically characterized large basket intrinsic neurons which deployed extensive intracortical axonal arborizations. These findings point to the preservation of intracortical inhibitory networks in the feline model of GM1 gangliosidosis, and to the possibility of abnormal integration of somadendritic inputs in ganglioside-laden pyramidal neurons. PMID- 2992704 TI - Development of stress-induced gastric lesions involves central adenosine A1 receptor stimulation. AB - When rats were exposed to immobilization stress for 1-12 h, gastric lesions did not occur at 1-6 h but did at 12 h of immobilization. Exogenous adenosine increased stress-induced gastric lesions, and dipyridamole, a blocker of adenosine uptake, potentiated the action of adenosine. The selective adenosine A1 receptor stimulants N6-cyclohexyl adenosine (CHA) and N6-(L-phenylisopropyl) adenosine (L-PIA) produced gastric lesions even in non-stressed state and markedly potentiated in dose- and time-dependent manner in stressed state. The stimulatory effect of N6-(D-phenylisopropyl) adenosine (D-PIA) on ulceration was weaker than that of CHA or L-PIA. Furthermore, intracerebral ventricular (i.c.v.) injection of adenosine or adenosine analogues produced the most rapid and most potent exacerbation of stress-induced gastric lesions relative to those induced with subcutaneous (s.c.) injection. The stress lesions enhanced by CHA were not affected by phentolamine, yohimbine, prazosin, naloxone and cholecystokinin (CCK8) but were inhibited by caffeine, clonidine, morphine and beta-endorphin. The inhibitory effect of clonidine was not antagonized by yohimbine or prazosin. The inhibition by morphine was selectively antagonized by exogenous CCK8 as well as naloxone. These results suggest that endogenous adenosine is tonically active in stress lesion formation which is modulated by opiate systems. Clonidine as well as caffeine may function as a purinoceptor antagonist, and it seems unlikely that the inhibitory effect of clonidine on stress ulcer is due to activation of alpha-adrenoceptors. PMID- 2992706 TI - Enhancement of hippocampal field potentials in rats exposed to a novel, complex environment. AB - The hippocampus plays a crucial role in place learning in rodents and also exhibits a long-term enhancement of synaptic strength and postsynaptic excitability following electrical stimulation of its principal afferents. In the present report we suggest that these two observations may be related, by demonstrating an increase in synaptic and postsynaptic field potential amplitudes resulting from exposure to a spatially complex environment. PMID- 2992705 TI - Evidence for GABA mediation of sympathetic inhibition evoked from midline medullary depressor sites. AB - GABA antagonists blocked, and diazepam potentiated, inhibition of spontaneous sympathetic activity elicited by electrical stimulation of classic midline medullary depressor sites. Picrotoxin often converted inhibitory effects of raphe stimulation into sympathoexcitatory responses. Serotonin antagonists blocked these sympathoexcitatory responses. The midline medullary raphe complex is heterogeneous in respect to autonomic function with sympathoinhibitory elements mediated at least in part by GABA and sympathoexcitatory pathways mediated by serotonin. PMID- 2992707 TI - Innervation of small intensely fluorescent cells in frog sympathetic ganglia. AB - Autonomic ganglia have a population of small intensely fluorescent (SIF) cells with unknown function. We have investigated the afferent innervation of SIF cells in bullfrog sympathetic ganglia using intracellular recordings and light microscopy of stained preganglionic axon terminals. Contrary to previous knowledge, bullfrog SIF cells do indeed receive functional synaptic input and this innervation is provided solely by the C-type preganglionic nerve fibers. PMID- 2992708 TI - Genotypic variation in striatal calmodulin content. AB - CBA/J and BALB/cJ mice have quantitative differences in the nigrostriatal projection. The number of nigral tyrosine hydroxylase reactive neurons, nigral and striatal tyrosine hydroxylase activity and the density of striatal D-2 dopamine receptors are all less in the CBA/J compared to the BALB/cJ mouse. An unrelated strain, the C57BL/6J, has a striatal D-2 dopamine receptor density that is intermediate to that of CBA/J and BALB/cJ mice. CBA/J mice also show deficits in the ability of brain monoaminergic receptor systems to develop supersensitivity. Calmodulin may participate in several striatal dopaminergic receptor mechanisms. Thus, striatal calmodulin was examined in CBA/J, C57BL/6J and BALB/cJ mice. Striatal calmodulin was greater in CBA/J mice than in C57BL/6J or BALB/cJ. In all three strains, cerebral cortical calmodulin was similar. The percent distribution of total striatal calmodulin between soluble and particulate fractions was similar in the three strains. Calcium redistributed soluble striatal calmodulin into the particulate fraction and EGTA shifted calmodulin from the particulate into the soluble fraction. The percent of total striatal calmodulin redistributed by either treatment was similar in all three strains. Gel filtration chromatography of heat-treated soluble extracts from CBA/J and BALB/cJ striatum was similar in elution pattern, although more calmodulin was observed in extracts from the CBA/J. Possible mechanisms for the strain differences in calmodulin are discussed along with their relationship to strain differences in striatal dopamine receptor subtypes. PMID- 2992709 TI - Effect of intraventricular administration of catechol estrogens on catecholamine content in various brain regions. AB - Male rats treated for 7 days with either 2-hydroxyestradiol, 4-hydroxyestradiol or 2-hydroxyestrone had significantly lower striatal dihydroxy phenylacetic acid (DOPAC) levels when compared to the control group. After 14 days of treatment, groups which were treated with estradiol, 2-hydroxyestradiol or 4 hydroxyestradiol had significantly higher striatal dopamine levels and had gained significantly less weight when compared to the control group. These results indicate that estradiol and the catechol estrogens may act to reduce the activity of striatal dopaminergic neurons. PMID- 2992710 TI - Intrinsic and extrinsic mechanisms involved in the cerebrovascular reaction elicited by immobilization stress in rabbits. AB - Variations in cerebral blood flow and partial pressures of oxygen and carbon dioxide (pO2, pCO2) were studied in rabbits during short-duration (1 min) immobilization stress. The techniques used enabled us to determine these variables locally in the caudate nucleus in a continuous, simultaneous and quantitative fashion. It could be shown that cerebral blood flow and arterial blood pressure increased in parallel immediately after inducing the stress reaction, and that pO2 increased further, indicating that cerebral oxygen supply is maintained by the hyperaemia. Previous administration of a beta-receptor blocker or of a cholinergic receptor blocker significantly diminished the cerebrovascular reaction to stress, inducing a decrease in pO2 during the reaction. Administration of both blockers nearly abolished the cerebral vasodilation studied. Previous administration of an alpha-receptor blocker enhanced the reactive hyperaemia. No disturbance of the blood-brain barrier could be observed in rabbits subjected to stress. Intravenous injection of adrenaline, as well as angiotensin II inducing similar increases in blood pressure, had no comparable effect on the blood flow. The conclusion is that in this model of anxiety, neurogenic mechanisms are involved in the provision of a sufficient oxygen supply to the brain. PMID- 2992711 TI - Quail neural crest cells transformed by Rous sarcoma virus can be established into differentiating permanent cell cultures. AB - Quail neural crest cells derived from the truncal neural primordium, infected in vitro by Rous sarcoma virus (RSV) in January 1978, were induced to multiply and have been established into permanent cultures. These cultures contain cells that differentiate into melanocytes, neuron-like cells and flat cells. About 50% of these different cell types are tetanus-toxin positive. Electrophysiological studies have shown that some cells can generate action potentials similar to those reported in quail neural crest primary cultures. Taken together these data show that the RSV-transformed quail neural crest permanent cultures are composed of stem cells which can differentiate into cell types specific for neural crest. PMID- 2992713 TI - Factors involved in expression of neuron-specific and non-neuronal enolase activity in developing chick brain and in primary cultures of chick neurons. AB - The effect of various factors affecting non-neuronal enolase (NNE) and neuron specific enolase (NSE) was investigated in developing brain of two different chick strains, in primary cultures of pure neurons and of mixed cultures of neuronal and glial cells. NNE and NSE activities reached their maximum at an earlier stage of brain development in the fast growing Hybro strain than in the Leghorn strain. In pure neurons cultured during 6 days, NNE was stimulated by hydrocortisone in presence or in absence of serum. Dibutyryl cyclic AMP (diBcAMP) stimulated NNE only in serum-free medium. NSE activity was increased by glial cell-conditioned medium in presence of serum and by removal of serum from the medium. Hydrocortisone and diBcAMP had no effect on NSE. In mixed cultures of neurons and glial cells both enolase activities were raised in absence of serum. Hydrocortisone and diBcAMP had no effect. Steroid hormones, insulin and serum albumin also modify both enolase activities in pure neurons and in mixed cultures of neurons and glial cells. Our results suggest that NNE and NSE are regulated separately by various factors involved in nerve cell maturation. PMID- 2992714 TI - Characteristics of benzodiazepine receptor binding in living cultures of mouse cerebral cortex at physiologic temperature. AB - Benzodiazepine (BDZ) receptor binding was assayed in situ in dispersed cultures of fetal mouse cerebral cortex at 37 degrees C and at 0 degrees C and compared. Contrary to data obtained from disrupted tissues, receptor binding at physiologic temperature was reduced only to 40% of that observed in the cold when intact tissues were used for assay. Neuronal (clonazepam-displaceable) receptor binding was essentially unaffected by changes in temperature; however, non-neuronal (Ro5 4864-displaceable) binding was reduced to 25% of that of 0 degrees C. A relatively high-affinity (Kd approximately 24 nM) as well as a low-affinity (Kd approximately 200 nM) binding site were identified; the numbers of binding sites were only modestly reduced at physiologic temperature and probably reflect a predominant reduction in non-neuronal sites. PMID- 2992712 TI - Addition of functional amiloride-sensitive components to the receptor membrane: a possible mechanism for altered taste responses during development. AB - The developmental increase in response sensitivity of rat peripheral taste afferents to NaCl and LiCl occurs concomitantly with an increase in sensitivity to the sodium ion transport blocker amiloride. Lingual application of amiloride had no effect on chorda tympani nerve taste responses to monochloride salts in early postnatal rats. However, amiloride suppressed NaCl and LiCl responses in proportion to the increased sensitivity to these stimuli during development in postweaning and adult rats without suppressing responses to NH4Cl and KCl. PMID- 2992715 TI - Screening for neuropeptide-metabolizing peptidases during the differentiation of chick embryo retina. AB - Chick retina was screened for neuropeptide-metabolizing peptidase activity during development using a kininase bioassay in which hydrolysis of any peptide bond of bradykinin (Arg1-Pro2-Pro3-Gly4-Phe5-Ser6-Pro7-Phe8-Arg9) leads to inactivation, combined with chromatographic bradykinin-product analysis. Bradykinin was degraded at a high rate, 6.1-26.6 mU/mg protein, by retina homogenates of all developmental stages. Kininase activity increased 2.3-fold from the 8th to the 18th embryonic day and 2-fold in the immediate posthatching period relative to the activity level at hatching. Bradykinin-product analysis, 57-113% recovery of the peptide fragments, indicated that kininase activity corresponded mostly to endopeptidase A- and to endopeptidase B-like activities (hydrolysis of Phe5-Ser6 and Pro7-Phe8 peptide bonds, respectively) and to angiotensin I-converting enzyme activity at all developmental stages. The data indicated that the relative amounts of these activities vary during retina differentiation. PMID- 2992716 TI - Presence of a novel epithelial antigen on rat cerebellar cell lines as detected by a monoclonal antibody. AB - We have derived a monoclonal antibody, MCAb 51, following immunization of BALB/c mice with a Rous sarcoma virus-transformed rat cerebellar cell line. When assayed by immunofluorescence on primary rat cerebellar cultures MCAb 51 recognizes only islands of cells with an epitheloid morphology. Double-label immunofluorescence experiments with MCAb 51 and antisera to tetanus toxin, glial fibrillary acidic protein, galactocerebroside and fibronectin reveal that these cells do not appear to be neurons, astrocytes, oligodendrocytes, or fibroblasts, respectively. In contrast, cells from kidney, liver, tongue and choroid plexus epithelium are positive for the antigen. Of 12 Rous sarcoma virus-transformed cell lines, in contrast to 2 out of 9 chemically transformed lines, 11 exhibit the MCAb 51 antigen. These findings demonstrate that MCAb 51 recognizes an epithelial cell surface marker. Possible explanations for the difference in the expression of the antigen on Rous sarcoma virus and chemically transformed neural lines are discussed. PMID- 2992717 TI - Characterization of angiotensin II binding sites on neuroblastoma X glioma hybrid cells. AB - Mouse neuroblastoma X rat glioma hybrid cells, NG108-15, have recently been shown to contain immunoreactive angiotensin II (AII). In the present study, we have examined this hybrid cell line for the presence of specific AII binding sites using [125I] AII. Specific AII binding was saturable and reversible. Scatchard analysis revealed a linear plot with an affinity constant (Kd) of 0.323 nM and a binding capacity (Bmax) of 7.13 fmol/mg protein. Kinetic studies demonstrated an association rate constant (K+1) of 3.55 X 10(6) M(-1) sec-1 and a dissociation rate constant (K-1) of 4.18 X 10(-4) sec-1. Displacement curves, using concentrations of 10(-11) M to 10(-4) M of unlabeled AII, revealed high and low affinity components of the AII binding site with IC50's of 0.46 nM and 1.75 microM respectively. The AII antagonist, saralasin, had approximately equal potency with unlabeled AII at the high affinity site. Furthermore, structurally related and unrelated peptides had no significant inhibitory effect on AII binding. This study demonstrates that specific AII binding sites are present on NG108-15 cells, and that these binding sites are similar in kinetic character to the AII receptor that has been previously identified in membranes from mammalian brain. It is concluded that the NG108-15 hybrid cells may provide a useful continuous cell line model for investigating both the biochemical and molecular properties of the AII binding site. PMID- 2992718 TI - The noradrenergic neurotoxins DSP4 and xylamine bind to opiate receptors. AB - DSP4 and xylamine compete with [3H]-naloxone for opiate binding sites with IC50 values of approximately 1 microM. This effect can be blocked by excess naloxone but not by the noradrenaline uptake inhibitors cocaine or desipramine. Other drugs containing the 2-chloroalkylamine structure--phenoxybenzamine, dibenamine, chloroethyl clonidine, the cholinotoxin AF-64 and 2-dimethylaminoethyl chloride- were similarly tested. Phenoxybenzamine and dibenamine were also able to compete with [3H]-naloxone for binding at the opiate receptor. Experiments in vivo demonstrated that DSP4, like other opiates, can rapidly reduce LH secretion in the rat. This effect is prevented by naloxone but not by desipramine. These data suggest the use of caution in interpreting the results of experiments in which DSP4 and xylamine are used as "specific" noradrenergic uptake inhibitors or as neurotoxins. PMID- 2992719 TI - Sham feeding, flavor associations and diet self-selection as indicators of feeding satiety or aversive effects of peptide hormones. AB - We have attempted to develop a constellation of behaviors which show differential effects following the administration of putative satiety hormones (CCK-8, BBS, insulin) as opposed to effects seen following a toxin, such as LiCl. In the initial behavior assessed, sham feeding of differently paired, flavored milks (flavor paired with insulin, BBS or saline) was carried out. Male adult Sprague Dawley rats which sham fed milk flavors paired with 16 micrograms/kg BBS showed a significant aversion of that flavor in a two-bottle taste test (compared to saline-paired flavors, p less than 0.001) but a significant preference for flavored milk paired with 0.4 and 0.75 U insulin/rat. Lower dosages of BBS (4 and 8 micrograms/kg) and insulin (0.1 U/rat) showed no significant aversion or preference when compared to saline. The second behavioral paradigm evaluated the effects of the hormones CCK-8 and BBS and the toxin, LiCl, upon self-selection of pure macronutrients. While CCK-8 reduced intake of calories by significantly lowering ingestion/selection of fats (55%, p less than 0.01 compared to saline, control injections) and carbohydrates (50%, p less than 0.01), LiCl and BBS reduced calories by decreasing selection of primarily proteins (LiCl--49%, p less than 0.03; BBS--63% at 4 micrograms/kg and 80% at 8 micrograms/kg, both p less than 0.025). In both paradigms then, BBS at doses sufficient to significantly reduce sham intake or suppress caloric ingestion in a self-selection paradigm produced behavioral effects most similar to those observed following the injection of a toxin. LiCl, rather than effects seen following other various putative satiety signals. PMID- 2992721 TI - Opioid systems and feeding in the slug, Limax maximus: similarities to and implications for mammalian feeding. AB - Substantial evidence is accumulating to implicate opioid systems in the regulation of behavioral and physiological functions in invertebrates in a manner analogous to that observed in vertebrates. This communication reviews opiate involvement in the mediation of the ingestive behaviors of the terrestrial slug, Limax maximus. The similarities to and implications for opioid modulation of mammalian feeding are considered. PMID- 2992722 TI - Visceral factors in the control of food intake. AB - Some years ago, we reported that the increased blood intake of hypoglycemic rats was inhibited by the intravenous infusion of fructose, a sugar that cannot cross the blood-brain barrier and nourish cerebral chemoreceptors. More recent experiments therefore have focused on visceral factors in the control of food intake. Three observations have been emphasized in this review. First, we found that gastric emptying was increased during insulin-induced hypoglycemia, and that this effect also was eliminated by administration of fructose. Hepatic vagotomy abolished both this effect of fructose on gastric emptying and its effect on food intake. Second, we found that in rats with severe diabetes, the rate of gastric emptying did decrease in proportion to increasing concentration of an administered glucose load, as it does in intact rats, but calories emptied more rapidly than normal regardless of the concentration of the load. Third, we found that rats with varying degrees of streptozotocin-induced damage to the pancreas ate more food than intact rats did after an overnight fast, and that individual intakes were proportional to the induced glucose intolerance. The increased eating took the form of shorter intermeal intervals, as if the initial postfast meal did not remain satiating for a normal amount of time. These and other findings suggest that food intake is controlled in part by satiety signals apparently related to the delivery of utilizable calories plus insulin to the liver. These signals also seem to affect gastric emptying and thereby might influence other satiety signals related to gastric distention. PMID- 2992720 TI - Opioids and consummatory behavior. AB - Since the second decade of this century it has been known that opiates can influence ingestive behaviors. Generally, opioid agents enhance feeding and opioid antagonists decrease feeding. The present paper reviews the responsiveness of different animal species to opiates in relation to ingestive behaviors, the opioid receptors involved in such consummatory behaviors, the site of action of opioid modulation of feeding, the role of glucose in opioid induced feeding, and endocrine effects on opioid feeding systems. We emphasize the finding that more than one opioid receptor is involved in the modulation of feeding. A large body of evidence indicates a major role for the dynorphin/alpha-neo-endorphin kappa opioid receptor as one of the receptors involved in feeding modulation. Opioids appear to exert their effect predominantly within the central nervous system, though peripheral effects on taste and gastrointestinal function may play a role in opioid-induced feeding. Although opioid blockade acutely blocks food intake, chronic administration of opiate antagonists to humans and laboratory animals has not proven to be an effective means of decreasing body weight. Chronic opiate administration decreases body weight and autosensitization of beta-endorphin increases body weight. Thus, although it is clear that opioids can effect food intake, it is not clear what effect chronic administration of opioids has no food intake or body weight. PMID- 2992723 TI - [Characteristics of the src transforming gene of viral and cell origin]. PMID- 2992724 TI - Continuous five-drug versus alternating three-drug and two-drug chemotherapy after five-drug or three-drug induction in extensive small cell lung cancer. AB - Fifty-six patients with extensive small cell lung cancer were treated with vincristine, doxorubicin, cisplatin, etoposide, and cyclophosphamide in a prospective randomized trial using all five drugs together or as three-drug (cisplatin, etoposide, cyclophosphamide) and two-drug (doxorubicin, vincristine) combinations given sequentially, then alternatively. The five-drug combination was associated with a higher overall regression rate (p = 0.03), higher complete regression rate (p = 0.09), prolonged time to first progression (p = 0.03), more nervous system initial failures (p = 0.07), more anemia requiring transfusion (p = 0.04), but no prolongation of overall survival (median 332 days for five-drug, median 303 days for three-drug and two-drug). Until more and better chemotherapeutic agents become available, little or no advantage is likely to be gained for patients with small cell lung cancer given sequential, or alternating chemotherapy. PMID- 2992725 TI - Controversies in treatment of small cell carcinoma of the lung. PMID- 2992726 TI - Herpes simplex virus latency: adaptation to the peripheral nervous system. II. PMID- 2992727 TI - Vitamin A, dehydroepiandrosterone (DHEA) and 5' nucleotidase: regulatory factors in tumor growth. PMID- 2992728 TI - [Specific cytotoxic activity of a virus-induced murine lymphoma]. AB - The NS8 lymphoma induced with Radiation Leukemia Virus (RadLV) presents in vitro a specific cytotoxic activity. This line was established from a lymphoma induced by injection of lymph node cells from mice bearing the MCA-fibrosarcoma T2 to congenic mice. These cells present a cytotoxic activity limited to the T2 cells. They develop no significant cytotoxic activity against other syngeneic or allogeneic tumoral lines. PMID- 2992729 TI - [Further studies on the biological characteristics of human colonic mucinous adenocarcinoma transplanted in nude mice]. PMID- 2992730 TI - Enflurane, halothane and isoflurane do not inhibit angiotensin converting enzyme activity. AB - We studied the effect of halothane, enflurane and isoflurane on angiotensin converting enzyme (ACE) activity using [3H]-benzoyl-phenylalanyl-alanyl-proline (BPAP) as a substrate. Isolated rabbit lungs were perfused in a recirculating system in vitro with BPAP in Krebs-Ringer solution. The rate of metabolism and per cent metabolism were determined before and after treatment for 30 minutes with four MAC multiples of enflurane, halothane or isoflurane. The effects of the anaesthetics on ACE activity were determined by calculating per cent inhibition of metabolism of BPAP using data from the control and test period for each lung. The average metabolism of BPAP at 15 minutes during the control period was 76.5 per cent (+/- 1.92 SEM). No anaesthetic significantly inhibited metabolism of BPAP. Likewise there was no effect on BPAP first order kinetics. Although potent inhalation anaesthetics may alter the renin-angiotensin-aldosterone axis, they do not affect this crucial step. PMID- 2992731 TI - Genetic relationship between pUB110 and antibiotic-resistant plasmids obtained from thermophilic bacilli. AB - The molecular relationship between pUB110 (Kmr, 4.4 kilobases (kb] and antibiotic resistant plasmids from thermophilic bacilli, pTHT15 (Tcr, 4.5 kb) and pTHN1 (Kmr, 4.8 kb), were studied by blot hybridization. Extensive homology was observed between pUB110 and pTHT15 at the region which includes the replication origin. Incompatibility studies revealed that pTHT15 and pUB110 were slightly incompatible in Bacillus subtilis but that they were apparently compatible in B. stearothermophilus. This difference in incompatibility between pTHT15 and pUB110 in the two host cells might be due to a difference in the copy number of pTHT15 in the two organisms. From the results of blot hybridization, mode of kanamycin inactivation, and DNA sequencing, it was determined that pTHN1 encoded the identical gene for kanamycin nucleotidyl transferase as that of pUB110. All three plasmids pTHT15, pTHN1, and pUB110 shared a common DNA homology at the in vitro membrane-binding region. PMID- 2992732 TI - Comparison of the mitochondrial endonucleases from Neurospora crassa and Saccharomyces cerevisiae. AB - The endonucleases from Neurospora crassa and Saccharomyces cerevisiae are not closely related antigenically. They also differ with respect to their activity at pH 8, their degree of hydrophobicity, and their sensitivity to elevated temperatures. However, the two nucleases have similar specific activities, are inhibited by EDTA, and have nearly identical substrate specificities. Since the enzymes also have the same mode of action and intracellular location, these similarities may indicate that they have the same physiological role despite their structural differences. PMID- 2992733 TI - Detection of antibody to HTLV-III by commercial kits. PMID- 2992734 TI - Psychiatric aftercare in a metropolitan setting. AB - In the face of the trend toward brief hospitalization, rising re-admission rates and other indices of poor community adjustment, concern has developed about the adequacy of psychiatric aftercare services. The authors report on a comprehensive study of psychiatric aftercare in a large metropolitan area (population 2.5 million). The study followed prospectively a group of 747 patients, representative of a significant proportion of patients in the care system. The findings document inadequacies in hospital-based discharge planning, unbalanced use of aftercare services and poor patient outcomes six months and two years post discharge. The study found heavy reliance on medical/therapeutic aftercare services with a relative neglect of housing, vocational/educational, financial and social/recreational services. Despite the large volume of medical/therapeutic service use, the patient group had a high readmission rate, high levels of symptomatology and poor social adjustment on follow-up. The authors suggest that community-based practitioners with specialized training in psychiatric rehabilitation would improve the system of aftercare. PMID- 2992735 TI - Testicular germ cell tumors. A review of 10 years' experience. AB - Experience with 200 cases of testicular tumors seen over a period of 10 years has been presented. Modes of presentation, response to treatment, and patterns of failure have been analyzed in 186 cases of germ cell tumors. The role of radiation in the overall management of seminomas is reaffirmed, and the need for adjuvant radiation to the supraclavicular region and mediastinum is demonstrated in Stage II cases of seminoma. Results of treatment with radiation alone or combined with chemotherapy were not encouraging in nonseminomas in the current series. The advantages of an aggressive multimodal approach with judicious combination of surgery, radiation treatment, and combination chemotherapy have been discussed, with review of relevant literature. PMID- 2992736 TI - Three months' treatment with cyclophosphamide, VP-16-213 followed by methotrexate and thoracic radiotherapy for small cell lung cancer. AB - One hundred eleven patients with inoperable but limited-stage small cell lung cancer were treated with three courses of cyclophosphamide (1.5, 2.5, and 3.5 g/m2, respectively) and VP-16-213 followed by methotrexate and thoracic radiotherapy. The total duration of treatment was 3 months. Patients were included who had pleural effusions, contralateral neck nodes, and bone marrow infiltration. The complete response (CR) rate was 56%, the majority confirmed by repeat bronchoscopy, with an 81% overall response rate. The minimum follow-up was 14 months. Median survival for all 111 patients was 11 months and 14 months (1 34+) for complete responders; the median survival was also 11 months for the 91 patients with conventional limited-stage disease, although 15 of the 19 patients alive at 14 months or more were from this subpopulation. There was no significant difference in the survival of those CR patients whose response was confirmed bronchoscopically and patients whose CR was assessed only radiologically and clinically. Forty-four patients with leukopenia (less than 1000 cells/microliter) received intravenous antibiotics for malaise and suspected infection. Close monitoring between treatments and direct access of patients to the hospital was encouraged. The majority of patients improved symptomatically as assessed by Karnofsky and Respiratory scores. These results support the view that short but intensive treatment without long-term or maintenance chemotherapy is beneficial. PMID- 2992737 TI - Insulinoma. An immunocytochemical and morphologic analysis of 95 cases. AB - One hundred twenty-seven insulinomas from 95 cases (1 malignant and 94 benign) were studied pathologically. Thirty-six tumors (35 cases) were examined by electron microscopy. Typical beta-cell secretory granules of crystalloid-form cores and/or atypical secretory granules were discerned in all tumors examined. A new type of secretory granule, with high electron-dense crystalloid-form cores and moderate electron-dense granular substance filling the space between the core and the limiting membrane, were observed in two cases. Among 68 insulinomas (67 cases) subjected to immunocytochemical investigations with ten peptide hormones (insulin, glucagon, somatostatin, pancreatic polypeptide (PP), gastrin, motilin, secretin, vasoactive intestinal polypeptide (VIP), gastric inhibitory polypeptide (GIP), and neurotensin), 42 were found to be multihormonal, varying from two to four peptides secreted. The hormones contained were insulin, glucagon, PP, somatostatin, and gastrin in different combinations. One patient had hyperinsulinemia and hypergastrinemia concurrently, and two islet tumors were excised at an interval of 10 months. Both electron microscopy and immunocytochemistry confirmed the presence of beta- and alpha-cells in the first tumor, whereas the second tumor revealed only G-cells by electron microscopy, and G- and beta-cells on immunocytochemical staining. The morphologic and immunocytochemical characteristics of the insulinomas in this series are discussed. PMID- 2992738 TI - The changing management of Wilms' tumor over a 30-year period. 1949-1978. AB - Seventy-four patients with histologically confirmed diagnoses of Wilms' tumor were treated between 1949 and 1978 at the Mallinckrodt Institute of Radiology and St. Louis Children's Hospital. The results have been divided into two eras of therapy, i.e., before and after 1965, when chemotherapy became a major modality for Wilms' tumor therapy. Analysis at 5 years by era of therapy has shown similar disease-free survival results for Stage I ("prechemotherapy" era 67% versus "chemotherapy" era 75%), whereas the Stage II ("prechemotherapy" era 33% versus "chemotherapy" era 100%) and Stage III ("prechemotherapy" era 0% versus "chemotherapy" era 70%) are significantly different (P less than 0.001). This suggests that chemotherapy has substituted for postoperative irradiation in Stage I patients; whereas in Stage II and III, improved survival occurs as a result of the eradication of subclinical metastatic disease. The patterns of failure by era of treatment are presented and the literature is discussed. PMID- 2992739 TI - Malignant fibrous histiocytoma of jaws. A clinicopathologic study of 11 cases. AB - Malignant fibrous histiocytoma (MFH) of the jaws is a highly malignant tumor that recurs, metastasizes, and usually causes death despite aggressive surgical therapy. This clinicopathologic review looks at five patients with MFH of the maxilla and six with MFH of the mandible. Five male and six female patients ranged in age from 12 to 75 years (mean, 35.4 years). All patients had large lytic areas of bone destruction, often with soft tissue extension. Two cases were postirradiation sarcomas, one of the maxilla and the other of the mandible. All patients underwent surgery and eight patients received chemotherapy when disease recurred locally or metastasized. Seven patients had local recurrences 3 to 13 months following surgery, and six patients had distant metastases. Of the 11 patients, 7 died of their disease, 1 died of unknown causes, and another with extensive local disease was lost to follow-up after 1.7 years. Two patients with recurrent disease are alive at 18 and 27 months postoperatively. PMID- 2992740 TI - Comparison between the nuclear diameters of primary and metastatic breast cancer cells obtained by cytologic aspiration. AB - In order to determine if there are morphologically identifiable characteristics between malignant cells obtained from a primary cancer and its metastasis the nuclear diameter was used as an indicator of the degree of malignancy, since there is good correlation between nuclear size, DNA content, and chromosome numbers. The nuclear diameter of primary and metastatic mammary carcinoma cells, obtained by cytologic aspirates, was measured by ocular micrometry. The purpose was to investigate whether a cell population at the primary site developed, at the metastatic sites, a population with the same nuclear size or one having larger and more anaplastic nuclei. One hundred eighty-five patients with infiltrating ductal carcinoma of the common variety were examined. The primary cancer and axillary nodal metastasis were examined in 97 patients before treatment. Thirty had cytologic examination of the breast cancer, as well as of the metastasis, which developed 1 to 14 years after treatment. Eleven were examined before radical breast irradiation and again at the time of relapse in the breast. Forty-seven had bilateral synchronous mammary carcinoma and both primary cancers were studied. The data presented indicate that there is extreme similarity between the nuclear diameters of the primary tumor and its metastasis. This similarity persists for several years regardless of both the location of the recurrence or radical irradiation. These results support the view that the majority of tumors are monoclonal in origin. The clone that invades the metastatic site appears to be the same as the one that initiated the primary cancer. In contrast, the nuclear diameters of cell populations obtained from synchronous bilateral breast cancer were dissimilar, indicating that they arose from separate clones of malignant cells. PMID- 2992741 TI - Large cell carcinoma of the lung. Ultrastructural differentiation and clinicopathologic correlations. AB - Light microscopic biopsy specimens from 48 patients were reviewed by two independent pathologists and classified as large cell carcinoma of the lung by 1981 World Health Organization (WHO) criteria. Sites of primary disease were hilar/mediastinal in 26 patients, large mid-lung field in 17, and peripheral lung in 5. All material was examined by electron microscopy (EM) for evidence of squamous ("squamous": 15 patients), glandular ("adenocarcinoma": 17 patients), or nonspecific ("large cell": 14 patients) ultrastructural differentiation. Two patients had mixed adenosquamous features. There were 6 patients with Stage I tumors; 5, Stage II; 24, Stage IIIM0; and 13, Stage IIIM1. Of the 14 patients with large cell by EM, 11 had unresectable Stage IIIM0 or metastatic disease. Only 3 of 27 patients not undergoing resection responded to combined modality therapy. There were two long-term survivors free of disease in the resected Stage IIIM0 patient category. Overall median survival by stage was analyzed, with no statistically significant difference between several of the stage groupings, suggesting a worse prognosis for the entire group overall compared to all patients with non-small cell lung cancer. The median survival by EM subgroup was also without significant difference, both overall and within various stage groupings, despite more patients in the large cell category with advanced disease. These data support the unique behavior of patients with large cell carcinoma on light microscopy, but fail to demonstrate that ultrastructural differentiation is of prognostic importance for response or survival. PMID- 2992742 TI - Large cell neuroendocrine tumors of the lung. Clinical significance and histopathologic definition. AB - Twenty-five cases of neuroendocrine tumors of lung, including bronchial carcinoids (eight), malignant (atypical) carcinoids (nine), and large cell undifferentiated carcinomas (LCAC) with neuroendocrine differentiation (eight) were analyzed. All carcinoids (BC) could be diagnosed by light microscopy; all patients with these tumors are alive without disease. Five of nine malignant carcinoids (MC) could be recognized histologically; four of nine were called LCAC and required electron microscopy for diagnosis. Survival correlated best with stage of diagnosis. High-grade neuroendocrine carcinomas (LCAC-NE) required electron microscopy for their diagnosis. Seven were LCAC histologically; one was diagnosed as malignant carcinoid (MC). Such tumors resemble small cell anaplastic carcinomas ultrastructurally and behaviorally. All eight patients with such high grade tumors died of their disease; three of eight had no nodal metastases at the time of resection. This experience suggests it is clinically important to distinguish neuroendocrine neoplasms since behavior is predictable on the basis of morphology. PMID- 2992743 TI - Pancreatic polypeptide in islet cell tumors. Morphologic and functional correlations. AB - Twelve islet cell tumors and one islet cell hyperplasia were studied with immunocytochemical and radioimmunoassay methods. With immunocytochemical staining, all six insulinomas, one mixed insulinoma-glucagonoma, and four gastrinomas were positive for insulin, insulin and glucagon, and gastrin, respectively. Pancreatic polypeptide (PP) was positive in three insulinomas and one mixed insulinoma-glucagonoma. All of the tumors were positive for neuron specific enolase (NSE). Radioimmunoassays of tissue extracts further disclosed that all functioning tumors contained more than one pancreatic hormone. PP concentrations of two insulinomas and one mixed insulinoma-glucagonoma were higher than that of normal control pancreases. A study of protein meal-stimulated PP secretion revealed that three of the insulinoma cases and two gastrinoma cases exhibited higher plasma PP levels than the age-matched controls. The findings suggest that: both functioning and nonfunctioning islet cell tumors derive from neuroendocrine cells positive for NSE; all functioning islet cell tumors appear to contain PP in the tumor tissue as a minor component; as many as 70% of the patients with islet cell tumors present with abnormally higher plasma PP levels after protein meals; and a study of meal-stimulated PP secretion may well be used as a marker for the presence of functional islet cell tumors. PMID- 2992744 TI - Concurrence of lymphoma type adult T-cell leukemia in three sisters. AB - Three sisters, ranging in age from 56 to 59 years, who developed lymphoma type adult T-cell leukemia (ATL) during a 19-month period are described. The patients were born in the Amakusa area of Kumamoto Prefecture, an area where the incidence of malignant lymphoma is high. The histologic diagnosis, made on the basis of the Lymphoma Study Group in Japan (LSG) classification of lymph nodes, was diffuse, medium-sized cell type in the elder sister, and mixed cell type in the middle and younger sisters. The patients and their elder brother had serum antibodies against ATL-associated antigens (ATLA). PMID- 2992745 TI - Cystosarcoma phyllodes. Report of an unusual case, with death due to intraneural extension to the central nervous system. AB - Cystosarcoma phyllodes tumors of the breast occasionally exhibit malignant behavior, including chest wall invasion, hematogenous spread or, rarely, metastasis by lymphatic routes. An unusual case of cystosarcoma is presented in which death was apparently caused by perineural and intraneural extension of the tumor along intercostal nerves, to the sympathetic chain, and then to the brain stem by spinal roots, with no evidence of embolic metastasis. PMID- 2992746 TI - Breast cancer with osseous metastasis and herniated lumbar disc. A cautionary tale. AB - A 39-year-old white woman with breast cancer, metastatic to her skeleton, developed low back and left lower extremity pain and lower extremity weakness. A bone scan evidenced increased radioisotope activity in her lumbar spine and a computed tomography (CT) scan showed a lesion of the L4 vertebra. Because of myelographic findings of a extradural defect at the L4-5 disc space and the possibility of a herniated disc causing the patient's pain and neurologic deficit in her lower extremities, the patient underwent surgery and a large herniated L4 5 disc was removed. As a consequence, the patient experienced relief of the lower extremity pain and return of strength in her lower extremities. She died a considerable time later from refractory hypercalcemia. PMID- 2992747 TI - Paraneoplasic precocious puberty. Report of a new case with hepatoblastoma and review of the literature. AB - A new case of precocious puberty secondary to production of human chorionic gonadotropin (HCG) by an hepatoblastoma was studied in an 8-month-old infant. A review was made of the 35 cases of paraneoplasic precocious puberty previously reported in the literature. The most frequent cause is hepatoblastoma, which was responsible for 18 cases. There are nine reports of mediastinal teratoma, six of which were observed in patients with Klinefelter Syndrome. In another six cases, the paraneoplasic syndrome was attributed to intracranial tumoration, and two of these patients showed teratoma with areas of choriocarcinoma, whereas germinoma was suspected for clinical reasons in another three. Lastly, one case of presacral teratoma and one of retroperitoneal carcinoma were reported. The differential characteristics of paraneoplasic precocious puberty are: almost exclusive occurrence in males, because of tumoral production of HCG, moderate or absent increase of testicle size, hyperplasia of Leydig's cells without spermatogenesis, and rapidly progressive signs of puberty. PMID- 2992749 TI - An appraisal of the World Health Organization classification of tumors of the central nervous system. AB - An important development in the classification of tumors of the central nervous system was the nomenclature recommended by the World Health Organization in 1979. Since then, wide usage has suggested that the classification be modified to account for problems that neuropathologists have encountered. Although no classification is ideal, modification of certain tumor categories, particularly from the perspective of childhood brain tumors, is desirable. These are discussed in detail. Greater emphasis on standardization of the localization of brain tumors is also recommended. This appraisal suggests changes that will allow a consensus on nomenclature of central nervous system tumors so that a workable classification is the end product. PMID- 2992748 TI - Tumor markers and cytologic features of cerebrospinal fluid. AB - Tumor markers are useful in establishing the diagnosis of certain central nervous system tumors, especially germinal tumors of the pineal region. They are not sufficiently specific to be able to replace biopsy for exact diagnosis. They may also be useful for monitoring of therapy, as an indicator of recurrence of the tumor. Cerebrospinal fluid cytology is not generally useful in establishing a specific histologic diagnosis, especially in children, but can help to monitor therapy and predict tumor recurrence. More extensive studies are needed in both areas to define more precisely the role of markers and cytologic studies. PMID- 2992750 TI - Revision of the World Health Organization classification of brain tumors for childhood brain tumors. AB - A classification for childhood brain tumors based upon revision of nomenclature of all brain tumors published by the World Health Organization (WHO) in 1979 is proposed. Applicability of the WHO classification scheme was tested in a combined study of the clinical and pathologic features of approximately 3300 brain tumors in children. It was found to be adequate for many of the neoplasms but unsuitable for a significant proportion, including a number of complex cerebral tumors for which there was no appropriate name. Nomenclature of poorly differentiated or densely cellular neuroepithelial tumors was simplified to reflect the current state of knowledge of neuroembryology and neuro-oncology, although the Committee members recognized that such a proposal would likely perpetuate the long-standing and continuing controversy relative to the nature and origin of these neoplasms. PMID- 2992751 TI - A commentary on the proposed revision of the World Health Organization classification of brain tumors for childhood brain tumors. PMID- 2992752 TI - Evaluation of drug efficacy in vitro using human small cell carcinoma of the lung spheroids. AB - Five human small cell carcinoma of the lung (SCCL) cell lines selected from 25 established cultures were grown as three-dimensional spheroid tumor models in either spinner culture or in static, agar-coated multiwells. Volume doubling times for the cell lines were approximately 4.5 days. Decreases in spheroid volumes after exposure to a variety of chemotherapeutic agents were used as indicators of drug activity. To further quantify cell killing in SCCL spheroids by chemotherapeutic agents 24 hours after exposure to drugs, a technique was employed that measured maximum levels of incorporation of 125IUdR after continuous labeling for 48 hours. The results of the use of this assay report for SCCL spheroid responses to various concentrations of doxorubicin hydrochloride, cytosine arabinoside, mechlorethamine hydrochloride, cisplatin, or etoposide. Some evidence for an intertumor heterogeneous response to chemotherapy is presented for some of the drugs tested. This assay was also used to characterize a potentiated cell kill when etoposide is combined with cisplatin and to identify activity by a new compound, diazoacetylcholine iodide (DACI), which was synthesized as an agent targeted for SCCL cells. PMID- 2992753 TI - Treatment of unresectable or metastatic osteosarcoma with cisplatin or cisplatin doxorubicin. AB - Responses and toxicity after treatment with cisplatin and cisplatin-doxorubicin were compared in two groups of patients with unresectable or metastatic osteosarcoma. Complete or partial responses developed in 3 of 18 individuals treated with cisplatin, and in 5 of 19 after the two-drug combination. Hematologic and gastrointestinal complications were more frequent and severe in patients who received both agents. The combined use of cisplatin and doxorubicin is justified for patients with unresectable or metastatic osteosarcoma at diagnosis because of the potential therapeutic benefits for these individuals. PMID- 2992754 TI - Heterogeneous estrogen receptor levels detected via multiple microsamples from individual breast cancers. AB - Twenty-six surgical breast cancer specimens were subjected to multiple estrogen receptor (ER) assays with a microsample technique. In most tumors there were large variations in ER levels in samples taken from different regions of the same tumor. After correction of individual samples for differences in carcinoma content, such variations were not abolished. Nine of the 26 breast cancers, although positive overall, were devoid of ER in some regions. For ER-positive tumors the average coefficient of variation (CV) for intratumor ER levels was 86%, with a range from 25% to 200%. This is well above the CV obtained with repeat samples of homogeneous tissue (14%). These results suggest that many ER positive cancers may be composed of cells with a variety of ER levels. An assessment of individual intratumor ER variability may have biologic and clinical significance. PMID- 2992755 TI - Prognostic significance of nuclear DNA content in small cell carcinoma of the lung. AB - The relationship between the nuclear DNA histogram patterns of tumor cells obtained by brushing via bronchoscopy and the survival time of 39 patients with small cell carcinoma of the lung was investigated. The brushing smears were stained by a modified Feulgen method. The nuclear DNA content was measured at 550 nm using a microspectrophotometer. The DNA histogram pattern was classified into type I, which contained a higher proportion of G0G1-phase cells, and type II, which contained a higher proportion of S, G2M-phase cells. The median survival times for the patients with limited disease of type I, type II and the patients with extensive disease of type I, type II were 17.2 months, 10.2 months, 10.0 months, and 5.6 months, respectively. Patients with limited disease of type I had a significantly longer survival time than patients with limited disease of type II. A distinct correlation was found between the histogram pattern and the survival of the patients. These results indicated that the nuclear DNA histogram pattern may be an indicator to allow speculation as to the prognosis of patients with small cell carcinoma of the lung. PMID- 2992756 TI - Bilaterality in familial breast cancer patients. AB - The authors conducted this study to determine the probability of a second primary developing in the contralateral breast of a patient, based on her present age, age at her first breast primary, and family history of breast cancer in her mother, sister, or second-degree relative. The study involved 556 patients. With premenopausal diagnosis of the first primary, the probabilities to the 19th year for the three types of family histories ranged from 35% to 38%; with postmenopausal diagnosis, they ranged from 11% to 26%. The probability for all familial patients was 28%. This contrasts with a probability of 13% reported for a general series of patients. An early age at diagnosis and family history of the disease thus have important enhancing effects on the development of second primaries. This information could be useful to physicians in deciding how best to manage the treatment of their patients. PMID- 2992757 TI - Presenting conditions of 1539 population-based lung cancer patients by cell type and stage in New Hampshire and Vermont. AB - The authors identified all newly diagnosed lung cancer cases in New Hampshire and Vermont for the period 1973 through 1976 and abstracted clinical data on presenting symptoms and findings from their hospital records. Microscopy slides were also reviewed, when possible, to confirm cell type. The most frequent presenting symptoms were weight loss (46%) and cough (45%). Other common symptoms were dyspnea (37%), weakness (34%), chest pain (27%), and hemoptysis (27%). The presence of symptoms and findings was in general related to disease stage but bore little relationship to cell type. These results differ from those of previously reported case series that were based on surgical, radiation therapy, or Veterans Hospital groups, but the current data agree closely with those from another population-based series in Finland. PMID- 2992758 TI - Disparity in the effects of two N-methyl nicotinamides on poly(ADP-ribose) synthetase and macromolecular synthesis in hepatomas. AB - The inhibitory effects of nicotinamide analogs on the activity of poly(ADP ribose)) synthetase were compared to effects on precursor incorporation into macromolecules in three lines of hepatoma cells (Morris hepatomas 5123C, 7777 and HTC). N'-methylnicotinamide was a less effective inhibitor of poly (ADP-ribose) synthetase than was 1-methylnicotinamide while both these compounds had smaller inhibitory effects on the enzyme than were seen with nicotinamide or 3 aminobenzamide. On the other hand, the incorporation of [3H]thymidine into DNA and of [3H]uridine into RNA were inhibited by N'-methylnicotinamide in the concentration range 2-20 mM but not by 1-methylnicotinamide. Under the conditions examined there were no significant effects on the incorporation of [14C]lysine and [3H]leucine in hepatoma cells. The data indicated that the inhibitory effect of N'-methylnicotinamide on nucleic acid synthesis may be unrelated to action on poly (ADP-ribose) synthetase. PMID- 2992759 TI - Electron spin resonance and high pressure liquid chromatographic analysis of melanin in posterior choroidal melanomas. AB - Electron spin resonance (ESR) and high pressure liquid chromatography (HPLC) were utilized to characterize the physical and biochemical characteristics of melanin in choroidal melanoma. ESR free radical signals indicative of eumelanin could be elicited from formalin-fixed paraffin-embedded tissues. Zinc ions (50 mM) increased the number of melanin-free radicals resulting in greater ESR sensitivity. Pyrole 2,3,6 tricarboxylic acid (PTCA) and amino hydroxy phenylalanine (AHP) were identified by HPLC after permanganate oxidation and hydroiodic acid hydrolysis, respectively, of a choroidal melanoma obtained at enucleation. The results indicate eumelanin is the primary melanin type in posterior choroidal melanomas. The feasibility of these techniques in the detection of metastatic disease from ocular melanomas is discussed. PMID- 2992760 TI - No easy answers. PMID- 2992761 TI - The PLC/PRF/5 human hepatoma cell line. I. Reevaluation of the karyotype. AB - The karyotype of the PLC/PRF/5 (Alexander) human hepatoma cell line was identified at passage +/- 110, prior to attempting in situ hybridization studies to determine the chromosomal localization of the hepatitis B virus (HBV) DNA integration sites. The karyotype was established by means of G-, Q-, and sequential C-banding techniques. Multiple consistent numerical and structural abnormalities were detected. These were compared with the original published unbanded karyotype of this cell line (at passage less than 25) and also with a previously published banded karyotype (at passages 60-90). Despite minor differences in the compared banded karyotypes (which are probably interpretational), the concordance in modal number and morphological karyotypic similarities over the course of 10 years indicate that this cell line is stable in vitro. PMID- 2992762 TI - The PLC/PRF/5 human hepatoma cell line. II. Chromosomal assignment of hepatitis B virus integration sites. AB - The chromosomal sites at which hepatitis B virus (HBV) DNA is integrated into the genome of the hepatocellular carcinoma (HCC) cell line, PLC/PRF/5 were investigated in an attempt to understand the mechanisms by which hepatitis B virus may induce malignant transformation. In situ hybridization of an HBV DNA probe to metaphase chromosomes of the PLC/PRF/5 cell line, followed by statistical analysis, identified three integration sites; these were 15q22-q23, 11q22, and 18q12. In particular, hybridization to chromosome #15, which is present in four copies in complete metaphases of this cell line, was highly significant (p much less than 0.0005). PMID- 2992763 TI - Sequence homologies in the control regions of c-myc, c-fos, HTLV and the interleukin-2 receptor. AB - Homologies in the control regions of 2 cellular oncogenes have been identified in this study. Both oncogenes (c-myc and c-fos) are known to be transiently induced by mitogens. We suggest that transcriptional activators bind to these putative control sequences, thus de-regulating gene expression in a coordinated and cell cycle specific manner. In addition, we report on homologous sequences in the control regions of the human T-cell leukemia viruses types I and II, and in the flanking region of the gene coding for the interleukin-2 receptor. These, and other experimental data, lead to the formation of a model in terms of which the unlimited proliferation of cells infected with HTLV-I and -II may be explained. The differing biological effects of HTLV-I, -II and -III are also examined and discussed at a molecular level. PMID- 2992764 TI - Caffeine and other phosphodiesterase inhibitors are potent inhibitors of the promotional effect of TPA on morphological transformation of hamster embryo cells. AB - The phosphodiesterase inhibitors caffeine, theophylline, aminophylline and isobutyl-methylxanthine (IBMX) were found to inhibit induction of morphologically transformed hamster embryo cell colonies by sequential exposure to benzo[a]pyrene (BaP) and the tumor promoter TPA. Almost complete inhibition of cell transformation was observed when 50 micrograms/ml theophylline, aminophylline, IBMX, or 200 micrograms/ml caffeine was present together with the tumor promoter. The compounds had no effect on the transformation frequency when present together with the initiator, BaP, in the first exposure period. Substances that stimulate the adenylate cyclase and the addition of exogenous dibutyryl-cAMP had similar inhibitory effects. PMID- 2992765 TI - Alterations in concentrations of cyclic guanosine 3',5'-monophosphate in guinea pig urine during the development of a transplantable leukaemia. AB - Cyclic guanosine 3',5'-monophosphate (cGMP) concentrations were measured in urine specimens from guinea pigs before and after transplantation of a leukemia. A transient increase in cGMP concentration occurred 3 days after inoculation which preceded any detectable increase in the white blood cell count. This peak in cGMP concentration was found to be very highly significant when compared with fluctuations in urinary cGMP levels in the guinea pigs from transplantation of the leukemia. Inoculation of guinea pigs with irradiated L2C cells suggests that transplantation may cause an initial depression in urine cGMP concentration but no elevated cGMP concentrations were observed in these animals. The results indicate that urine cGMP concentrations may be a useful way of monitoring abnormal cell proliferation. PMID- 2992766 TI - Macrophages and metastasis--a biological approach to cancer therapy. PMID- 2992767 TI - Prolonged stimulation of S91 melanoma tyrosinase by [Nle4, D-Phe7]-substituted alpha-melanotropins. AB - alpha-Melanotropin (alpha-melanocyte stimulating hormone, alpha-MSH) stimulates tyrosinase activity in Cloudman S91 murine melanoma cells. Three [Nle4, D-Phe7] substituted alpha-melanotropin analogues, [Nle4, D-Phe7]-alpha-MSH, Ac-[Nle4, D Phe7]-alpha-MSH4-11-NH2, and Ac-[Nle4, D-Phe7]-alpha-MSH4-10-NH2, are at least 100-fold more effective than alpha-MSH in stimulating melanoma tyrosinase, the rate-limiting enzyme in melanin biosynthesis. These [Nle4, D-Phe7]-substituted melanotropin analogues induce tyrosinase activity in melanoma cells with shorter contact times than required by the native hormone, alpha-MSH. [Nle4, D-Phe7] substituted melanotropins also induce a prolonged (residual) stimulation of melanoma tyrosinase. Following incubation of melanoma cells in the presence of [Nle4, D-Phe7]-alpha-MSH for 24 h, tyrosinase activity is maintained for up to 6 days in the absence of the melanotropin. The shorter 4-10 and 4-11 fragment analogues also exhibit residual melanotropic activity. The prolonged stimulation of tyrosinase in the absence of the analogues is maintained even though melanoma cells continue to divide about every 24 h. These results suggest that melanoma cells possess spare melanotropin receptors and that [Nle4, D-Phe7]-substituted analogues bind almost irreversibly to these receptors or to some other component of the adenylate cyclase enzyme complex responsible for enhancing tyrosinase activity and melanin production. PMID- 2992768 TI - Calcitonin stimulation of cyclic adenosine 3':5'-monophosphate production with growth inhibition in human renal adenocarcinoma cell lines. AB - Responsiveness of cyclic adenosine 3':5'-monophosphate (cAMP) to parathyroid hormone, calcitonin, and vasopressin was studied in six human renal adenocarcinoma cell lines. Four of six renal adenocarcinoma cell lines showed increased cAMP content in response to calcitonin while the other two did not. Neither parathyroid hormone nor vasopressin increased the concentration of cAMP in each of these cell lines. The growth rate of KU-2 cells, which responded to calcitonin with an increase of cAMP content, was inhibited by calcitonin. On the other hand the growth rate of calcitonin-nonsensitive KH-39 cells was unaltered. The growth inhibitory effect of the hormone on KU-2 cells could be considered to be mediated by the increased cAMP levels from the following results: (a) there was positive correlation between the cellular cAMP content and growth inhibition after various amounts of calcitonin addition; (b) KU-2 growth was also suppressed by N6,O2'-dibutyryl cAMP; and (c) a group of KU-2 cells which had become resistant to calcitonin-induced growth inhibition showed a diminished cAMP increase in response to calcitonin. PMID- 2992769 TI - Cell surface antigens of chemically induced fibrosarcomas: detection by a monoclonal antibody of a tumor-restricted Mr 12,000 protein gag antigen encoded by a dual-tropic murine leukemia virus. AB - Fusion products of spleen cells of W/FuDp rats immunized with a methylcholanthrene-induced BALB/c sarcoma, CA-2, and mouse myeloma cells were screened in an attempt to identify a monoclonal antibody defining the individually distinct tumor-specific transplantation antigen of CA-2. A hybridoma, MP/69/04, was isolated which produces an IgG2a monoclonal antibody that recognized a tumor-restricted antigen of CA-2. In direct binding assay, MP/69/04 reacted only with 2 of 15 methylcholanthrene induced BALB/c sarcomas tested. Thymus, spleen, lymph nodes, bone marrow, brain, adult lung fibroblasts, newborn muscle fibroblasts and 3T3 cells were negative. Absorption tests revealed, however, expression of the MP/69/04 determinant on 8 of the 12 murine leukemia virus (MuLV) producer BALB/c sarcoma tested. The antigen was not detected on any of the three non-producer sarcomas tested nor on a wide range of normal tissues and cell lines. An N-dualtropic MuLV was isolated from CA-2, and cell lines susceptible to infection by this virus were shown to express the MP/69/04 epitope. By Western blotting, the MP/69/04 epitope was identified as being expressed on the MuLV structural protein with a molecular weight of 12,000, present in CA-2 cells and in the purified CA-2 MuLV. These results indicate the MP/69/04 antigen is not a unique tumor-specific transplantation antigen but is a gag product of a recombinant retrovirus which is expressed on the cell surface of many MuLV + methylcholanthrene-induced BALB/c fibrosarcomas. PMID- 2992770 TI - Inhibition of hepatic phosphoenolpyruvate carboxykinase by avian reticuloendotheliosis viruses. AB - Severe weight loss is associated with many malignant diseases of humans and animals. Avian reticuloendotheliosis viruses (RE viruses) induce runting in experimentally infected chickens. Chickens infected with a replication-competent RE virus, reticuloendotheliosis-associated virus, weighed 30-50% less than control birds at the time of death. Chickens infected with reticuloendotheliosis virus, a replication-defective acute leukemia virus, weighed 30% less than the controls. The runting induced by RE viruses does not occur because of reduced food intake. Activities of phosphoenolpyruvate carboxykinase, a key gluconeogenic enzyme in the liver, were reduced approximately 40 and 50%, respectively, by infection with reticuloendotheliosis-associated virus and reticuloendotheliosis virus. RE virus infection, however, did not affect the hepatic pyruvate carboxylase activity, indicating that inhibition of phosphoenolpyruvate carboxykinase is not due to a general inhibition of all liver enzymes. Birds given injections of UV-inactivated RE viruses or reticuloendotheliosis virus transformed, non-virus-producing tumor cells also exhibited a reduction in phosphoenolpyruvate carboxykinase activity. PMID- 2992771 TI - Inhibitory effects of dipyridamole on growth, nucleoside incorporation, and platelet-activating capability in the U87MG and SKNMC human tumor cell lines. AB - The effects of dipyridamole on tumor cell function were examined in cultures of two lines of human origin, the SKNMC neuroblastoma line that activates platelets by a mechanism which is dependent on the release of adenosine 5'-diphosphate and the U87MG glioblastoma line that induces platelet activation by the generation of thrombin. Cells grown in the presence of dipyridamole at 1 microM showed greater than 80% inhibition of uptake of adenosine, thymidine, and uridine with both lines. At 5 microM tumor cell growth was inhibited by 70% (U87MG) and 90% (SKNMC) but without concomitant cytotoxicity as determined by clonogenic assay (50% inhibitory concentration approximately 20 microM). At 10 microM dipyridamole cyclic adenosine 3':5'-monophosphate levels increased 150% with both cell lines but no changes above baseline values were seen at 2.5 microM. The two cell lines showed different responses to being cultured in the presence of dipyridamole in terms of their ability to subsequently activate platelets. U87MG cells cultured in 10 microM dipyridamole showed a doubling of the lag time as compared with cells grown in the absence of dipyridamole but with full aggregation; with SKNMC cells the aggregation rate was reduced and cells grown in 10 microM dipyridamole showed no reversible first wave, a 5-fold increase in lag time and a 75% inhibition in total aggregation. Since therapeutic doses of dipyridamole result in plasma concentrations of approximately 3.5 microM these results suggest that potential antimetastatic effects of dipyridamole could be direct arising from inhibition of important steps in tumor cell metabolism or indirect by suppressing one or more of the mechanisms involved in the ability of tumor cells to activate platelets. PMID- 2992772 TI - Oncodevelopmental enzymes of the Dunning rat prostatic adenocarcinoma. AB - In this paper, data are presented which demonstrate that adenylate kinase and creatine kinase are oncodevelopmental enzymes in the rat prostate. The Dunning tumor (dorsal rat prostate) was used as a model system; four sublines of the tumor (R3327-H, R3327-AT, MAT Lu, and MAT LyLu) were studied. The tumor lines were maintained as solid tumors in syngeneic rats (Copenhagen) and as monolayers in tissue culture. The appearance of adenylate kinase with malignant transformation of the dorsal prostate was demonstrated. The disappearance of the CK-M subunit of creatine kinase and decreasing levels of creatine kinase were demonstrated with increasing anaplasia. The lactate dehydrogenase (LDH) concentration increased with increasing anaplasia, and the LDH isoenzyme pattern shifted to a more glycolytic pattern (LDH-4, LDH-5). The malignant isoenzyme pattern was reversible with the use of a differentiating agent (dimethyl sulfoxide). Prostates from neonatal rats and castrated adult male rats exhibited patterns of creatine kinase and adenylate kinase similar to those of the undifferentiated tumor. The oncofetal isoenzyme pattern of the castrated rat prostate was reversible with physiological levels of exogenous testosterone. PMID- 2992773 TI - Effects of disulfiram, diethyldithiocarbamate, bisethylxanthogen, and benzyl isothiocyanate on glutathione transferase activities in mouse organs. AB - Four sulfur compounds known to inhibit tumorigenic effects of chemical carcinogens were administered to female CD-1 mice at 0.5% of the diet for 14 days, and their effects on cytosolic glutathione transferase (EC 2.5.1.18) specific activities were examined in liver, lung, kidney, urinary bladder, forestomach, proximal small intestine, and colon. Disulfiram, sodium diethyldithiocarbamate, bisethylxanthogen, and benzyl isothiocyanate elevated glutathione transferase specific activities in most of the organs examined. The four sulfur compounds differed in the extents and organ specificities of their effects on these enzyme activities. In the liver, bisethylxanthogen and benzyl isothiocyanate increased glutathione transferase activities to at least 3 times control levels and caused differential increases in the isozyme patterns observed after isoelectric focusing of the cytosols. Bisethylxanthogen also increased immunoreactive glutathione transferase in liver cytosol. Recrystallized disulfiram was less effective in enhancing hepatic glutathione transferase activities than was commercial (97%) disulfiram. Among the six extrahepatic organs examined, the small intestine and the forestomach exhibited the greatest response of glutathione transferase activities to each of the four sulfur compounds. Benzyl isothiocyanate was most effective in these "portal of entry" organs but less effective than bisethylxanthogen in the other extrahepatic organs examined. Bisethylxanthogen elicited significant increases in glutathione transferase activities in liver, lung, and small intestine even when administered at 0.01% to 0.05% of the diet, suggesting that this compound may have considerable potential as an inhibitor of carcinogens susceptible to enzymatic conjugation with glutathione. PMID- 2992774 TI - Effect of poly(adenosine diphosphate-ribose) polymerase inhibitors on neocarzinostatin-induced G2 delay in HeLa-S3 cells. AB - The antitumor antibiotic neocarzinostatin (NCS), which produces single-strand breaks in mammalian cell DNA in vivo, stimulated the activity of chromatin bound enzyme, poly(ADP-ribose) polymerase in HeLa-S3 cells. Because of the possible causal relationship between the poly ADP-ribosylation of chromatin protein and NCS-induced temporary G2 arrest in the cell cycle, several classes of inhibitors of poly(ADP-ribose) polymerase were examined to evaluate the effect on NCS induced polymerase activity as well as on progression in the cell cycle of synchronized HeLa cells which had been treated with NCS in G2. Compared at the same concentration of 2 mM, the polymerase-inhibiting activity was larger in the order of thymidine, 3-aminobenzamide, nicotinamide, theophylline, and caffeine. Among these agents, caffeine, theophylline, and thymidine caused a reduction in the G2 delay in this order by stimulating the cells to undergo mitosis after NCS treatment. However, 3-aminobenzamide and nicotinamide were poor reducers, if any, of NCS-induced G2 delay. These results suggest that there is not a direct involvement of poly ADP-ribosylation of chromatin protein in the mechanism of NCS induced G2 delay. The effect of caffeine on G2 delay will probably be independent of its activity as a poly(ADP-ribose) polymerase inhibitor. PMID- 2992775 TI - Effects of 9-OH-ellipticine on cell survival, macromolecular syntheses, and cell cycle progression in sensitive and resistant Chinese hamster lung cells. AB - In an effort to understand the mechanism of action of the DNA-intercalating antitumor agent 9-hydroxyellipticine (9-OH-E), we have examined the effects of this drug on the cell survival, macromolecular syntheses, and cell cycle progression in sensitive and resistant cells. Our results show that 9-OH-E toxicity on sensitive and resistant cells involves different mechanisms of action: the drug toxicity in the sensitive cells appears to result from lethal lesions mediated through the interaction of the drug with an intracellular protein, independently of any effect of the drug on the macromolecular syntheses; in the resistant cells, the cell death occurs concomitantly with the inhibition of these syntheses. Cell cycle progression analysis after 9-OH-E treatment showed that, in the sensitive cells, the drug is inducing a G1 and a G2 block, which are both released in the presence of 1 mM caffeine, without any effect on the 9-OH-E toxicity. In the resistant cells, a G2 block was also observed but only when the cells were resuming their growth after about a 30- to 40-h growth arrest. Caffeine release of this block, which again had no effect on 9-OH-E toxicity, was only observed when it was added from 40 to 60 h after 9-OH-E treatment, when the cells resumed their growth. Finally in the sensitive cells, cycloheximide exerted an inhibitory effect on 9-OH-E toxicity when it was added before and during the cell exposure to the drug. This effect was interpreted as indicating that 9-OH-E toxicity in the sensitive cells relies on a protein which is not induced by the drug but has to be present in the cells when the drug is added. The possible implication of DNA topoisomerases in 9-OH-E toxicity mechanism is discussed. PMID- 2992776 TI - Alternative differentiation of human promyelocytic leukemia cells (HL-60) induced selectively by retinoic acid and 1 alpha,25-dihydroxyvitamin D3. AB - Induction of hematopoietic differentiation was investigated in human promyelocytic leukemia cells [HL-60] using two lipophilic vitamins, retinoic acid and 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3]. Both vitamins suppressed proliferation and induced differentiation of HL-60 cells, but 1 alpha,25(OH)2D3 was 70- to 100-fold more potent than was retinoic acid on a molar basis. Simultaneous treatment with suboptimal concentrations of 1 alpha,25(OH)2D3 (0.12 to 1.2 nM) and retinoic acid (10 to 100 nM) showed additive effects in reducing nitroblue tetrazolium, a common marker for monocyte-macrophage and granulocyte differentiation. For the study of alternative differentiation of the cells by the two vitamins, we used monoclonal antibodies specific for either human monocyte macrophages or granulocytes and other markers specific for macrophage differentiation such as alpha-naphthyl acetate esterase activity and adherence to the dish surface. HL-60 cells were induced to differentiate alternatively into macrophages by 1 alpha,25(OH)2D3 or into granulocytes by retinoic acid. When HL 60 cells were treated with various concentrations of 1 alpha,25(OH)2D3 (1.2 to 120 nM) in the presence of 1000 nM retinoic acid which is a concentration sufficient to induce maximal granulocyte differentiation, the appearance of the markers for monocyte-macrophage differentiation by 1 alpha,25(OH)2D3 was not at all affected by the retinoic acid. These results indicate that 1 alpha,25(OH)2D3 and retinoic acid have additive effects in inducing differentiation of HL-60 cells, but monocyte-macrophage differentiation by 1 alpha,25(OH)2D3 occurs much more readily than does granulocyte differentiation by retinoic acid. PMID- 2992777 TI - Tumorigenicity and transcriptional modulation of c-myc and N-ras oncogenes in a human hepatoma cell line. AB - Tumorigenicity and oncogene expression were examined in HepG2 derived cells, a human hepatoma cell line. HepG2 cells and a single cell clonal HepG2 line, HLD2 6, were equally tumorigenic when injected s.c. into athymic nude mice. Cyclophosphamide pretreatment of both cell lines (500 micrograms cyclophosphamide/ml/two cell cycles) had no effect on tumor incidence or latency (P greater than 0.05). Tumors were nonencapsulated, highly invasive adenocarcinomas and were positive for gamma-glutamyltranspeptidase activity and bile production. Plasma from tumor-bearing mice was positive for human alpha fetoprotein and negative for hepatitis B virus surface antigen as measured by radioimmunoassay. Two cell lines reestablished into tissue culture from HLD2-6 derived tumors had unaltered cell cycle times. Detailed in vitro translation analysis of RNA isolated from HLD2-6 derived cells and tumors were extremely similar to the translation products of RNA isolated from a normal human liver sample except for a Mr 53,000 polypeptide with an apparent charge shift. c-myc specific transcripts, when compared to a normal human liver sample, were increased in all HLD2-6 cell lines and tumors derived from HLD2-6 cells. This increase in c-myc expression could not be explained by gene amplification or hepatitis B virus integration. N-ras specific transcripts were not elevated in HLD2-6 cells grown in tissue culture but there was a selective increase of the 5.5-kilobase N-ras transcript in HLD2-6 derived tumors grown in nude mice. This increased 5.5-kilobase transcript did not remain elevated if the tumors were reestablished into tissue culture, suggesting some interaction with the host animal. c-Ha-ras expression could not be detected in any HLD2-6 derived tumor or cell line. PMID- 2992778 TI - A variant F9 embryonal carcinoma cell line which undergoes incomplete differentiation in retinoic acid. AB - F9 embryonal carcinoma cells treated with 1 microM retinoic acid undergo irreversible differentiation and simultaneously lose their tumorigenicity. Described here are the isolation and characterization of an F9 variant clone (5C), which undergoes partial differentiation in retinoic acid. The behavior of 5C cells indicates that retinoic acid successfully initiates the differentiation pathway but that complete differentiation is not achieved due to a subsequent block in the pathway. The fact that 5C cells do not undergo complete differentiation, or lose their tumorigenicity in response to retinoic acid, indicates that the lesion affects an element involved in the regulation of both these events. Therefore, further genetic and biochemical characterization of this variant should provide information concerning the relationship between the regulation of differentiation and tumorigenicity. Furthermore, the isolation of this variant establishes the feasibility of genetically dissecting the various steps of the differentiation pathway. PMID- 2992779 TI - Use of agarose culture to measure the effect of transforming growth factor beta and epidermal growth factor on rabbit articular chondrocytes. AB - Nonneoplastic cell lines are unable to grow in soft agar. However, concomitant treatment of these cells with epidermal growth factor and transforming growth factor beta confers upon them anchorage independence. Since articular chondrocytes are unique as normal diploid cells that do have the capability of growing in soft agar, we tested whether transforming growth factor beta and epidermal growth factor could affect DNA synthesis and matrix production. In the presence of epidermal growth factor (5 ng/ml) concentrations of high-performance liquid chromatography-purified transforming growth factor beta at concentrations of 0.05-15 ng/ml induced a dose-dependent increase in DNA, to nearly double that of control cultures. A half-maximal effect was seen with transforming growth factor beta, 0.1 ng/ml, and epidermal growth factor, 5 ng/ml. Neither compound alone was mitogenic. In contrast, either transforming growth factor beta or epidermal growth factor alone was able to decrease synthesis of glycosaminoglycans and collagen. The data demonstrate that transforming growth factors can affect the behavior of nonneoplastic cells by modulating cell replication and the biosynthesis of two principal matrix components. In addition they support the hypothesis that these growth factors may play a role in the physiology of nonmalignant cells. PMID- 2992780 TI - Immune reactivity to Thomsen-Friedenreich antigen in patients with lung cancer detected by superoxide assay-leukocyte adherence inhibition test. AB - The immune reactivity to partially purified Thomsen-Friedenreich antigen was investigated in patients with lung cancer. The modified method of original leukocyte adherence inhibition test, termed superoxide assay-leukocyte adherence inhibition test, was used to detect the reactivity. The coded peripheral mononuclear cells from 34 of 50 (68%) patients with lung cancer showed a positive response to the antigen whereas in only 3 of 19 (16%) patients with benign pulmonary disease was there a reaction to the antigen. The same experiments were performed using the 3 m KCl extract of lung tumors as an antigen. In this case in 39 of 50 (78%) patients with lung cancer but in only 4 of 24 (17%) with benign pulmonary disease and in none of the breast cancer patients (0 of 17) was there a reaction to the antigen. These results strongly suggest that patients with lung cancer are sensitized to both Thomsen-Friedenreich antigen and tumor-associated antigens expressed in cancer cells of lung tissue origin. PMID- 2992782 TI - Brachial plexus neuropathy following high-dose cytarabine in acute monoblastic leukemia. AB - We describe brachial plexus neuropathy with high-dose cytarabine (Ara-C) therapy in a man who had acute monoblastic leukemia. Signs and symptoms of brachial plexus neuropathy appeared on two occasions within hours of exposure to high-dose Ara-C. Central nervous system complications have been described following systemic and intrathecal Ara-C. High-dose Ara-C has not been implicated previously as a cause of brachial plexus neuropathy. PMID- 2992781 TI - Characterization of the gamma-aminobutyric acid receptor system in human brain gliomas. AB - The properties of [3H]-gamma-aminobutyric acid [( 3H]GABA) binding were studied in biopsied specimens from normal human brain and from 18 cases of human brain gliomas, made up of 6 astrocytomas, 6 glioblastomas, 3 oligodendrogliomas, and 3 medulloblastomas. In fresh membranes obtained from normal gray and white matter one population of Na+-dependent GABA receptors was observed, while in the frozen Triton X-100-treated membranes two distinct populations of Na+-independent binding sites were detected. Specific GABA binding sites in brain gliomas were shown only in frozen Triton X-100-treated membranes. As in normal tissue, these receptors are Na+-independent and bind [3H]GABA with two distinct affinity components. The biochemical profiles of [3H]GABA binding to membranes obtained from different tumors of glial origin are quite similar and cannot be related to the degree of malignancy of the neoplasia. PMID- 2992783 TI - Intensive induction treatment of small cell bronchogenic carcinoma with cyclophosphamide, methotrexate, and etoposide. AB - Fifty-six patients with histologically confirmed small cell bronchogenic carcinoma were treated with cyclophosphamide, methotrexate, and etoposide. While methotrexate doses were modified for mucositis during the 6-week induction period, none of the drug doses were modified for hematologic toxicity. The overall response rate was 66%, with 16% complete remissions; median survival duration was 28 weeks. In 12 patients, the leukocyte count fell below 1000/mm3, and there were four deaths in febrile, leukopenic patients. Diffuse pulmonary infiltrates were observed in eight patients, and three died from respiratory insufficiency. Lung biopsy in two patients and autopsy in two additional patients showed interstitial changes consistent with drug injury. This regimen produced considerable hematologic and apparent pulmonary toxicity while offering no advantage in response rate or survival duration. PMID- 2992785 TI - Comparison of vindesine plus cisplatin or vindesine plus mitomycin in the treatment of advanced non-small cell lung cancer. AB - Fifty-eight patients with advanced non-small cell lung cancer were randomly allocated to receive vindesine (3 mg/m2 every week) plus either cisplatin (80 mg/m2 every 3 weeks) or mitomycin (8 mg/m2 weekly X 3, then every 3 weeks). No patients achieved complete response. Among the 28 patients treated with vindesine plus cisplatin, there were 12 partial responders (42.9%); among the 30 patients treated with vindesine plus mitomycin, there were only three partial responders (10%) (P less than 0.005). The median duration of response was 11.5 weeks (range, 4-25) in the patients treated with vindesine plus cisplatin. The median survival times for patients treated with vindesine plus cisplatin and vindesine plus mitomycin were 10.1 and 10.2 months, respectively; there was no statistical difference in survival time between the two groups. Initial performance status was the strong predictor of patient survival. Toxic effects, including moderate myelosuppression, nephrotoxicity, peripheral neuropathy, and gastrointestinal symptoms, were generally manageable. The combination of vindesine and cisplatin appears to be effective against advanced non-small cell lung cancer. PMID- 2992784 TI - Lomustine, vincristine, and procarbazine in the treatment of metastatic malignant melanoma. AB - Sixty-five previously untreated patients with metastatic malignant melanoma were treated with lomustine, vincristine, and procarbazine. Sixty-four patients were evaluable for response, with a response rate of 13%. Only one complete response was observed, in a patient with nodal disease only. Three partial responses were observed in patients with disease confined to soft tissue, and four partial responses were observed in patients with pulmonary metastases. Median survival for all patients was 22 weeks. We conclude that this regimen offers no improvement compared to other drug combinations. PMID- 2992786 TI - The effects of parathyroid hormone, dibutyryl cyclic AMP, calcitonin and colchicine on glucosamine incorporation into bone cells. PMID- 2992789 TI - [Changes in enzyme activity in cartilage and subchondral bone in experimental osteoarthrosis]. PMID- 2992787 TI - A possible mechanism of morphometric changes in dendritic spines induced by stimulation. AB - A number of experimental procedures which induce increased electrical activity (including long-term potentiation) were shown to be accompanied by morphometric changes in dendritic spines. These changes include an enlargement of the spine head, shortening and widening of the spine stalk, and an increase in the length of synaptic apposition. A possible mechanism is suggested which takes into account specific cytological features of the spine and the existence of contractile proteins in neurons. Dendritic spines are defined as special domains of the neuron which have a unique organization of the cytoplasm. Actin filaments form a very dense network in the spine head, and they are longitudinally organized within the spine stalk. Spines were also shown to contain myosin and other actin-regulatory proteins. The high density of the actin network could explain the characteristic absence of the cytoplasmic organelles from dendritic spines. In analogy with other cells, such an actin organization indicates low levels of free cytosolic calcium. Even in the resting state, calcium levels may be unevenly distributed through the neuron, being lowest within the subplasmalemmal region. Due to the high surface-to-volume ratio in spines, the cytoplasm is formed mostly by the subplasmalemmal region. The spine apparatus or the smooth endoplasmic reticulum, which is recognized as a calcium-sequestering site in spines, may also contribute to the low calcium levels there. However, when in the stimulated spine the voltage-dependent calcium channels open, then, given the spine's high surface-to-volume ratio, the concentration of calcium may very quickly attain levels that will activate the actin-regulatory proteins and myosin and thus trigger the chain of events leading to the enlargement of the spine head and to the contraction (i.e., widening and shortening) of the spine stalk. The increased free cytosolic calcium may also activate the protein producing system localized at the base of the spine, which, under certain conditions, could stabilize the morphometric changes of the spine. PMID- 2992790 TI - [The significance of angiotensin-converting enzyme in sarcoidosis]. PMID- 2992788 TI - Formation of retinotopic connections: selective stabilization by an activity dependent mechanism. AB - During regeneration of the optic nerve in goldfish, the ingrowing retinal fibers successfully seek out their correct places in the overall retinotopic projection on the tectum. Chemospecific cell-surface interactions appear to be sufficient to organize only a crude retinotopic map on the tectum during regeneration. Precise retinotopic ordering appears to be achieved via an activity-dependent stabilization of appropriate synapses and is based upon the correlated activity of neighboring ganglion cells of the same receptive-field type in the retina. Four treatments have been found to block the sharpening process: (a) blocking the activity of the ganglion cells with intraocular tetrodotoxin (TTX), (b) rearing in total darkness, (c) correlating the activation of all ganglion cells via stroboscopic illumination and (d) blocking retinotectal synaptic transmission with alpha-bungarotoxin (alphaBTX). These experiments support a role for correlated visually driven activity in sharpening the diffuse projection and suggest that this correlated activity interacts within the postsynaptic cells, probably through the summation of excitatory postsynaptic potentials (EPSPs). Other experiments support the concept that effective synapses are stabilized: a local postsynaptic block of transmission causes a local disruption in the retinotectal map. The changes that occur during this disruption suggest that each arbor can move to maximize its synaptic efficacy. In development, initial retinotectal projections are often diffuse and may undergo a similar activity dependent sharpening. Indirect retinotectal maps, as well as auditory maps, appear to be brought into register with the direct retinotopic projections by promoting the convergence of contacts with correlated activity. A similar mechanism may drive both the formation of ocular dominance patches in fish tectum and kitten visual cortex and the segregation of different receptive-field types in the lateral geniculate nucleus. Activity-dependent synaptic stabilization may therefore be a general mechanism whereby the diffuse projections of early development are brought to the precise, mature level of organization. PMID- 2992791 TI - [Prospective study of lung cancer in hospitalized patients. II. Radiographic picture]. PMID- 2992792 TI - Postnatal growth and differentiation in three hindlimb muscles of the rat. Characterization with biochemical and enzyme-histochemical methods. AB - The postnatal development, between 0 and 90 days, of three hindlimb muscles and diaphragm of the rat was investigated with respect to fiber types and diameter (histochemistry) and substrate oxidation rates and enzyme activities (biochemistry). The process of muscle fiber differentiation into mature patterns was evaluated by visual classification into 3 or 4 groups having different staining intensities for 3 enzyme-histochemical reactions, enabling 26 fiber types to be distinguished. These exhibited specific sizes and growth rates that varied among the muscles. One of the hindleg muscles (flexor digitorum brevis) remained much more immature than soleus and extensor digitorum longus. The histochemical and biochemical findings correlated well. The capacity for pyruvate and palmitate oxidation, and the activities of cytochrome c oxidase and citrate synthase, increased markedly between 9 and 37 days in soleus and extensor digitorum longus (except citrate synthase in the latter) but not in flexor digitorum brevis. Creatine kinase activity increased in all hindlimb muscles. Both the capacity and the activity of pyruvate oxidation (determined in homogenates and intact isolated muscles, respectively), were in accordance with the fiber type composition. In contrast to oxidation capacity, the activity of pyruvate oxidation decreased after birth until the mature stage, when a value of 18-42% of that of early postnatal muscles was recorded. PMID- 2992794 TI - The presence of vasoactive intestinal polypeptide (VIP)-like-immunoreactive nerve fibres and VIP-receptors in the pineal gland of the Mongolian gerbil (Meriones unguiculatus). An immunohistochemical and receptor-autoradiographic study. AB - By use of the indirect peroxidase-antiperoxidase immunohistochemical technique, nerve terminals exhibiting vasoactive intestinal polypeptide (VIP)-like immunoreactivity were demonstrated in the pineal gland of the Mongolian gerbil (Meriones unguiculatus). Incubation of the superficial pineal gland of the gerbil with 60 pM of 125I-VIP showed that the gland exhibited saturation kinetics, and about 80% of the bound 125I-VIP could be displaced by adding a surplus of cold VIP. Incubation of unfixed, 0.1% and 4% paraformaldehyde-fixed cryostat sections of the gerbil forebrain with 125I-VIP also exhibited saturation kinetics, and displacement was possible by adding a surplus of the cold tracer. Receptor autoradiography on cryostat sections that had been incubated for 60 min with 125I VIP showed a large number of grains over cortical areas, especially over the pyramidal layer of the hippocampus and the granular layer of the dentate gyrus. A prominent labelling of the pineal gland was also observed. The presence of VIP like-immunoreactive nerve terminals and receptors for this molecule in the pineal gland of the Mongolian gerbil supports the biochemical studies demonstrating a stimulatory function of this molecule in the synthesis of melatonin. PMID- 2992793 TI - Inosine diphosphatase as a histochemical marker of retinal microvasculature, with special reference to transformation of microglia. AB - Nucleoside diphosphatase (IDPase), localized using inosine diphosphate as substrate, allows the selective staining of blood vessels and cells of vascular origin, such as macrophages and microglia, whereas the neuroglial, the neuronal and the pigment epithelial cells remain unstained. The staining pattern observed in the retina of mouse, rat, cat and monkey are similar; some apparent quantitative differences reflect species differences in the distribution of retinal microvasculature. At the electron-microscopic level, most of the enzyme activity in the blood vessels appears to be located along the outer wall. The cell membrane, parts of the smooth endoplasmic reticulum and the nuclear membrane in the microglial perikarya appear positive; profiles of microglial processes are intensely stained. In the developing eyes of rats and mice, the blood vessels are stainable from the earliest stage of their appearance. An array of amoeboid cells precede the growing blood vessels and spread out over the future vascularized part of the retina. These cells eventually develop characteristic microglial features, and extend many elongated and branched processes between the neuroepithelial cells while remaining in contact with, or in close proximity to, the blood vessels. Intense IDPase activity in the microglial cells, in contrast to the absence of the enzyme in the neuroglial Muller cells, suggests that microglia are involved in phosphate metabolism and indicates functional compartmentalization within the glial tissue lying between the blood retinal barrier and the retinal neurons. PMID- 2992795 TI - Quantitative assessment of GABA-uptake sites in the neural lobe by electron microscopic autoradiography. AB - Distribution of (3H)GABA in the rat neural lobe was investigated 5 min after intracarotid administration using quantitative electron-microscopic autoradiography. Specificity of (3H)GABA-uptake was tested by pretreatment of control animals with nipecotic acid. It was concluded that, apart from a small fraction in the perivascular spaces, radioactivity was present exclusively in pituicytes. The results confirm and quantify earlier in-vitro observations; they are compared with recent immunocytochemical findings that reveal the presence of glutamate-decarboxylase-containing axons in the neural lobe. It is concluded that there may be GABAergic terminals that lack an uptake mechanism for exogenous transmitter. Nevertheless, (3H)GABA autoradiography is useful in demonstrating other functional components of GABAergic systems, i.e., glial cells. PMID- 2992796 TI - Functional and morphological characterization of cultures of Kupffer cells and liver endothelial cells prepared by means of density separation in Percoll, and selective substrate adherence. AB - This paper presents a study on the structure and function of Kupffer cells (KC) and liver endothelial cells (LEC) isolated by a simple and rapid technique involving 1) perfusion of the liver with collagenase; 2) cell separation by means of density centrifugation in Percoll; and 3) cell culture, taking advantage of the fact that KC and LEC differ in their preferences for growth substrate. The KC, which attach and spread under serum-free conditions on surfaces of glass or plastic during the first 15 min in culture exhibit a typical macrophage-like morphology including membrane ruffling and a heterogenous content of vacuoles. Moreover, these cells express (a) Fc receptors (FcR) for binding and phagocytosis of erythrocytes covered with immune globulin G (E-IgG), and (b) complement receptors (CR) for binding and serum dependent phagocytosis of erythrocytes covered with either human C3b or mouse inactivated C3b (iC3b). The cells also bind fluid phase fluoresceinated C3b. Approximately 30% of the KC express immune response-associated (Ia)-antigens. The LEC attach and spread on fibronectin coated surfaces, but not on glass or plastic surfaces, during the first two hours in culture with or without serum, and are morphologically distinct from KC. Cultured LEC are well spread out with no membrane ruffling and with numerous large vesicles surrounding the regularly shaped nucleus. These cells bind, but do not ingest E-IgG via the FcR, but no binding of fluid phase C3b or particle fixed C3b or iC3b can be observed. Incubation of LEC with fluorescein amine conjugates of ovalbumin or formaldehyde treated serum albumin, but not with fluoresceinated native serum albumin, results in accumulation of fluorescence specifically localized in the large perinuclear vesicles. Neither KC nor any other cell types tested have the ability to accumulate fluorescence upon incubation with these compounds. Ia-antigens are not present on the LEC. Cytochemical demonstration of unspecific esterase, acid phosphatase, and peroxidase reveals different patterns and intensities of staining in KC as compared to LEC. PMID- 2992797 TI - Mammalian collagenase predisposes bone surfaces to osteoclastic resorption. AB - The cell-free endocranial surface of young adult rat parietal bones was used as a substrate for bone cell-derived mammalian collagenase. Incubation of parietal bones in a concentration of enzyme comparable to that secreted by osteoblastic cells in vitro caused destruction of surface osteoid, and resulted in exposure of mineral onto the bone surface. Bones so pre-treated were considerably more susceptible to osteoclastic resorption than bones pre-incubated in the absence of collagenase. These results are consistent with the view that the osteoid layer which covers bone surfaces acts as a barrier to osteoclastic contact with underlying, resorption-stimulating bone mineral; and that cells of the osteoblastic lineage induce osteoclastic resorption through collagenase secretion which, by digestion of the surface osteoid, exposes bone mineral to osteoclastic contact. PMID- 2992798 TI - Intramembranous response to cAMP in fetal epidermis. AB - Prekeratinized fetal epidermis may function as an osmoregulatory organ. This study shows that the structural response of fetal epidermis to cAMP (second messenger of antidiuretic hormones) is very similar to that in well characterized osmoregulatory epithelia. cAMP stimulation of unkeratinized fetal epidermis induces the appearance of "aggregates" of intramembranous particles. These aggregates are present in the apical plasma membrane of the periderm or outermost cell layer of the epidermis. Tubular vesicles that contain the presumptive aggregates fuse with the apical plasma membrane during cAMP stimulation. The same response to cAMP and to antidiuretic hormones has been reported in the osmoregulatory ion- and water-transporting epithelia of amphibian urinary bladder and mammalian collecting ducts. In these systems aggregates have been positively correlated with water transport. Thus, the fetal epidermis may control and regulate its water permeability. PMID- 2992799 TI - The molecular genetics of hemophilia: blood clotting factors VIII and IX. PMID- 2992800 TI - Phosphoinositides in mitogenesis: neomycin inhibits thrombin-stimulated phosphoinositide turnover and initiation of cell proliferation. AB - Thrombin stimulates 32Pi incorporation into phosphatidylinositol 4-phosphate (PIP), phosphatidylinositol 4,5-bis-phosphate (PIP2), and phosphatidylinositol (PI), and initiates DNA synthesis in hamster (NIL) fibroblasts at a half-maximal concentration of 125 ng/ml. Neomycin, which binds PIP2 and PIP, inhibits both thrombin-stimulated initiation of cell proliferation and 32P pI incorporation into at concentrations above 2 mM without affecting thrombin binding, thymidine uptake, or cellular protein synthesis. At lower concentrations, neomycin inhibits thrombin-stimulated release of inositol 1,4,5-trisphosphate (IP3), by selectively binding PIP2, but does not inhibit 32P incorporation into PI or initiation of DNA synthesis. Phosphoinositide recycling and diacylglycerol release therefore appear necessary for initiation of cell proliferation by thrombin. IP3-stimulated Ca++ mobilization may not be required for thrombin mitogenesis, however, since neomycin can block IP3 release without inhibiting initiation. PMID- 2992801 TI - Translocation of secretory proteins across the microsomal membrane occurs through an environment accessible to aqueous perturbants. AB - We have characterized the association of a nascent secretory protein with the microsomal membrane at two distinct stages in cell-free synthesis and translocation. Stage one corresponded to a nascent chain of approximately 70 residues generated via elongation arrest by the signal recognition particle (SRP). Binding to microsomal membranes occurred independently of chain elongation and required SRP receptor. Following binding, the 70-mer remained attached to the membrane after extraction of the ribosome. However, protein denaturants (4 M urea or alkaline pH) extracted the 70-mer from the membrane. Stage two of synthesis corresponded to nascent chains of approximately 158 residues generated by oligonucleotide-mediated hybrid arrest of translation. Again, these partially translocated nascent chains were extracted by 4 M urea. Therefore, the initial interaction of the signal sequence with the membrane as well as subsequent chain conductance occur in a microenvironment that is accessible to aqueous reagents. Thus, both processes probably require integral membrane proteins. PMID- 2992802 TI - Negative regulation of viral enhancers in undifferentiated embryonic stem cells. AB - Many viral genomes, including those of SV40 and MuLV, are not efficiently expressed in undifferentiated embryonal carcinoma (EC) cells but are expressed in differentiated derivatives. This regulation appears to be at the level of transcription. We have used DNA-mediated gene transfer to analyze the function of several viral promoters in EC cells. We show that the SV40 early promoter works efficiently in an enhancer-independent fashion following transfection into undifferentiated cells. Strikingly, the promoter in the LTR of MSV does not function in such cells; but when upstream sequences, including the enhancer, are deleted expression ensues. Replacement of the SV40 enhancer by that of MSV results in inactivation of the SV40 early promoter in these cells. We propose that the undifferentiated cells contain a trans-acting regulatory factor (or factors) that reduces transcription by interacting with viral enhancers. PMID- 2992803 TI - Seventeen base pairs of region I encode a novel tripartite binding signal for SV40 T antigen. AB - Three sequence components direct high affinity binding of dimeric SV40 T antigen to SV40 origin region I. Two signals are encoded by two directly repeated 5' GAGGC-3' pentanucleotides. Approximately equal contributions to binding stability are made by each pentanucleotide, and both spacing and orientation of the pentanucleotides are important for binding affinity. The third vital component is contained in a 5'-TTTTTTG-3' spacer sequence that separates the pentanucleotides. Sequence-specific features of the spacer stabilize binding to the adjacent pentanucleotides. The asymmetry of the spacer suggests that a novel binding mechanism is involved. Because the alignment of T antigen on mutant and wild-type DNAs is similar, we propose that any two of the three sequence signals are sufficient to determine the unique arrangement of a bound protein dimer. PMID- 2992804 TI - Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro. AB - We have characterized an in vitro transcription system derived from uninfected HeLa cells that accurately initiates RNA synthesis at the herpes virus thymidine kinase (TK) promoter. Analysis of linker-scanning, single-site, and promoter inversion mutants reveals that the TK upstream elements previously mapped in vivo are accurately recognized in vitro. A protein fraction required for TK transcription in reconstitution experiments was found to contain multiple protein species that bind specifically to the TK promoter. DNAase I footprint experiments with wild-type and mutant promoters reveal that the TK upstream elements contain three distinctive protein binding sites, two of which appear to be recognized by the Sp1 transcription factor and one which interacts with a cellular protein that binds to "CCAAT" sequences. Optimal expression of the thymidine kinase gene appears to require the coordinate interaction of these two types of transcription factors with the three upstream elements of the promoter. PMID- 2992805 TI - Activation of a novel human transforming gene, ret, by DNA rearrangement. AB - A novel transforming gene was detected by transfection of NIH 3T3 cells with human lymphoma DNA. The tumor DNA induced a single focus in primary transfections, whereas DNAs of transformed NIH cells induced transformation with high efficiencies in secondary and tertiary assays. Molecular clones spanning about 37 kb of human sequence were isolated from tertiary transformant DNA. Blot hybridization indicated that the transforming gene consisted of two segments that were unlinked in both normal human and primary lymphoma DNAs. The two segments of human DNA were cotranscribed in transformed NIH cells but not in any human cells examined. The transforming gene thus appeared to be activated by recombination between two unlinked human DNA segments, possibly by cointegration during transfection. PMID- 2992806 TI - Structure of the eukaryotic transcription apparatus: features of the gene for the largest subunit of Drosophila RNA polymerase II. AB - The Drosophila melanogaster RpII215 locus encodes the largest subunit of RNA polymerase II. We have now mapped the 7 kb transcript of the locus and have determined that it contains four exons and three introns. By sequencing 2582 nucleotides from the promoter-proximal end of the RpII215 locus, we have precisely mapped the start site of transcription and the splice sites of the first intron. Segments of the amino acid sequence predicted by the only long open reading frame of the RpII215 gene transcript display striking homology with corresponding segments of the beta subunit of E. coli RNA polymerase. PMID- 2992807 TI - tnpM: a novel regulatory gene that enhances Tn21 transposition and suppresses cointegrate resolution. AB - We have identified a new gene, tnpM, in Tn21 that encodes the 12.6 kilodalton modulator protein. The Tn21 modulator enhances Tn21 transposition and suppresses resolution of cointegrate replicons in vivo. A putative binding site may be located in the N-terminal portion of the TnpR (resolvase) structural gene sequences. Tn501 transposition and cointegrate resolution can be regulated by the subcloned tnpM gene of Tn21 in trans-complementation experiments. Examination of the Tn501 DNA sequence also reveals a potential tnpM coding sequence upstream of the Tn501 resolvase gene. We conclude that Tn21 and Tn501 are different from Tn3 and Tn1000 both in genome organization and in regulation of transposition functions. PMID- 2992808 TI - B-cell-derived interleukin 1 (IL-1)-like factor. I. Relationship of production of IL-1-like factor to accessory cell function of Epstein-Barr virus-transformed human B-lymphoblast lines. AB - The relationship of production of interleukin 1 (IL-1)-like factor to accessory function of Epstein-Barr virus (EBV)-transformed B lymphocytes was examined. Six of eight human EBV-B cell lines spontaneously produced and released detectable levels of thymocyte comitogenic factor in vitro, but no interleukin 2 (IL-2) activity. Eight of eight produced fibroblast proliferation activity. Culture supernatants from the two apparent nonproducers of thymocyte comitogenic activity induced the proliferation of the IL-1-dependent murine helper-T-cell clone D10G4.1 in the presence of concanavalin A (Con A). One of the EBV-B cell lines produced a potent inhibitory factor in addition to IL-1-like thymocyte comitogenic and fibroblast proliferation factors. The inhibitory factor inhibited mouse thymocyte proliferative response to Con A, and the proliferation of the IL 2-dependent CT6 cell line, but not human fibroblast growth. All but one of the eight EBV-B cell lines tested, the exception being the line that produced an inhibitory factor, were able to serve as antigen-presenting cells that enabled purified human T lymphocytes to proliferate in one-way mixed lymphocyte reactions (MLR) and in response to Con A. The supernatants of 14 of 16 clones derived from two of the EBV-B cell line cells contained thymocyte comitogenic activity and all 16 stimulated fibroblast proliferation. The phenotypic characteristics of the EBV B cell lines were heterogeneous, but there was no clear-cut relationship between the cell surface phenotypes of either the cloned or uncloned EBV-B cells and their ability to produce these factors. These studies show that all of the EBV-B cell lines that can function as accessory cells have the capacity to produce an IL-1-like factor. PMID- 2992809 TI - B-cell-derived interleukin-1 (IL-1)-like factor. II. Sources, effects, and biochemical properties. AB - A variety of types of human B-cell lines were evaluated for their ability to produce interleukin 1 (IL-1)-like factors. All of the eight Epstein-Barr virus (EBV)-transformed B lymphocyte lines, three of four of the EBV+ lymphoma lines, only three of seven of the EBV- lymphoma lines, and none of the three tested myeloma lines secreted some IL-1 activity. The IL-1-like factor produced by the cell lines was detected on the basis of its thymocyte comitogenic and/or fibroblast proliferative activities. Injections of partially purified IL-1-like factor from one of the EBV-transformed B-lymphocyte lines also induced the appearance of an acute phase protein (haptoglobin) in the serum of C3H/HeJ mice. These biological activities are identical with those of monocyte-derived IL-1. Thymocyte comitogenic activity and fibroblast proliferation activity from one of the EBV-B cell line-derived IL-1-like activities were not dissociable by biochemical procedures, including HPLC gel filtration and HPLC anion-exchange chromatography. However, the IL-1-like factor from one of the EBV-B lymphocyte cell lines was larger in size (25 kDa) and more acidic (pI 5.5) than monocyte derived IL-1. PMID- 2992810 TI - Generation of a soluble IFN-gamma inducer by oxidation of galactose residues on macrophages. AB - Depletion of macrophages from human peripheral blood mononuclear cells (PBMC) caused a marked decrease in galactose oxidase and sodium periodate, but not a calcium ionophore, stimulated Interferon-gamma (IFN-gamma) production. Reconstitution of such depleted cultures with galactose oxidase treated macrophages, but not lymphocytes, restored IFN-gamma levels to those of control nonfractionated PBMC. Thus, galactose oxidase seemed to act on macrophages which in turn stimulated lymphocyte production of IFN-gamma. Unlike human cells which have terminal galactose residues on glycoproteins, murine cell glycoproteins terminate their oligosaccharide component in the order N-acetyl-neuraminic acid followed by D-galactose, N-acetyl-glucosamine, and glycoprotein. Galactose oxidase or sodium periodate only activated murine macrophages to stimulate lymphocyte IFN-gamma production after exposing D-galactose residues by the removal of the terminal N-acetyl-neuraminic acid residues with neuraminidase. Removal of such exposed terminal galactose residues with beta-galactosidase inhibited the effect of galactose oxidase on murine macrophages. Taken together, these results strongly suggest that oxidation of terminal galactose residues on macrophages is the initial site of action of galactose oxidase and sodium periodate. Studies with Boyden chambers have shown that galactose oxidase-treated macrophages released a soluble factor which stimulates lymphocyte production of IFN-gamma. Based on these findings, it appears that the oxidation of terminal galactose residues on the surface of macrophages leads to the induction and transmission of a soluble signal for lymphocyte production of IFN-gamma. PMID- 2992811 TI - Killer cells of feline leukemia virus- and feline sarcoma virus-infected transformed cells: the role of NK, ADCC, and in vitro generated cytotoxic cells. AB - Feline white blood cells (WBC) manifested a primary in vitro mixed lymphocyte tumor cell culture (MLTC) proliferative response to feline leukemia virus-feline sarcoma virus (FeLV-FeSV)-infected transformed target cells, which reached a peak at Day 15. Furthermore, primary in vitro MLTC cultures generated cytotoxic killer cells capable of killing a variety of targets in non-major histocompatibility gene complex restricted fashion, and effector cells were capable of killing targets introduced repeatedly into cultures over a 49-day period. The presence of feline fibrosarcoma (f-sarc) stimulators was the primary driving force for proliferation and generation of killing because exogenous IL-2 conditioned medium did not appreciably increase the yield of killer cells generated in vitro. WBC cultured without f-sarc stimulators with or without IL-2 supplementation also generated killer (K) cells but at a low level. The killer cell population was composed of approximately 50% lymphocytes and 50% monocytes. Cats had K cells functional in antibody-dependent cell-mediated cytotoxicity against FeLV-coated chicken red blood cells but not against any FeLV-FeSV-infected transformed targets tested. Natural killer (NK) cell activity to any targets tested was not found. Although no evidence was found for K or NK cell activity against FeLV-FeSV infected transformed cells, feline WBC were readily able to generate killer cells in vitro and it is probable that this cell-mediated immune potential is functionally important in vivo. PMID- 2992812 TI - Correlation between chemoattractant-induced leukocyte adherence inhibition, macrophage chemotaxis, and macrophage inflammatory responses in vivo. AB - Variations in the magnitude of inflammatory macrophage response in vivo and macrophage chemotaxis in vitro, observed among inbred mouse strains, suggest that these traits are genetically-regulated. The development of an A X B series of recombinant inbred (RI) strains of mice derived from the C57BL/6J (B, high responder) and A/J (A, low responder) resulted in the availability of a large number of new inbred strains which express a spectrum of variations in the magnitude of these traits. These strains were used in the present study as a tool to examine the possible correlation between the phenomenon of leukocyte adherence inhibition (LAI) and those of macrophage inflammatory response in vivo and macrophage chemotaxis in vitro under the assumption the LAI requires the same cellular events as chemotaxis and that LAI resembles, grossly, the accumulation of nonadherent inflammatory cells in vivo. The typing of A X B RI strains for the traits of LAI, macrophage accumulation in vitro, and macrophage inflammatory response in vivo resulted in a correlation between the magnitude of response of those three phenomena in the total of 19 inbred strains tested, thus suggesting that the chemoattractant-induced LAI is biologically related to the events that mediate macrophage chemotaxis in vitro and the macrophage inflammatory response to sterile irritants in vivo. PMID- 2992813 TI - Effects of granuloma modulation induced by regulatory-T-lymphocyte activity on angiotensin II/III production by granuloma macrophages in murine schistosomiasis mansoni. AB - Angiotensins are produced by granuloma macrophages in murine Schistosoma mansoni. During the course of infection, granuloma undergo a T-cell-dependent process called modulation in which their maximal size decreases. This study was undertaken to establish whether angiotensin production by granuloma macrophages is altered by immunoregulatory lymphocytes. Granuloma macrophages from modulated lesions released and contained more angiotensin II/III (AII/III) and less angiotensin I (AI) than those from the acute infection. Captopril, a specific angiotensin-converting-enzyme (ACE) inhibitor, appreciably decreased AII/III produced by macrophages from modulated granulomas. Adoptive transfer of splenic T lymphocytes from chronically infected donors into acutely infected recipients altered angiotensin production by the granuloma macrophages in a manner similar to that seen in modulated lesions. However, no difference was detected in the capacity of granuloma macrophages from acutely or chronically infected mice to metabolize 125I-AI or -AII added to cell cultures. Similarly, captopril did not alter the metabolism of exogenously administrated angiotensins. These findings suggest that regulatory T lymphocytes influence the metabolism by granuloma macrophages of endogenously produced angiotensins at least in part by induction of macrophage ACE activity. However, the degradation of extracellular AI and AII may result from the activity of enzymes other than ACE which are not inducible by modulation. PMID- 2992814 TI - IgM rheumatoid factor autoantibody and immunoglobulin-producing precursor cells in the bone marrow of humans. AB - The natures of the IgM rheumatoid factor (RF)-, IgM-, and IgG-secreting cells in the human bone marrow as compared to the peripheral blood, have been investigated by (1) response to the polyclonal B-cell activator, the Epstein-Barr virus (EBV), (2) sensitivity to the S-phase specific antimetabolite hydroxyurea, (3) presence of the BA-1 and Ia antigens on the cell surface, and (4) cell size, as determined by counter flow elutriation. The EBV-inducible bone marrow IgM-RF precursors derived from medium to large B cells that were inhibited by hydroxyurea pretreatment. The marrow total IgM response derived from small to medium size cells, and was only partially inhibited by hydroxyurea. Hydroxyurea had no effect on IgM-RF or IgM synthesis by peripheral blood cells. These results indicate that the marrow EBV-induced IgM-RF response is not representative of the response by peripheral blood cells, moreover; the marrow RF secreting response arises from a dividing cell pool that may represent newly generated autoreactive B cells. PMID- 2992815 TI - Interferon-gamma is a strong modulator of NK susceptibility and expression of beta 2-microglobulin but not of transferrin receptors of K562 cells. AB - The human cell line K562 was treated with human natural leukocyte interferon (IFN alpha) and recombinant immune interferon (IFN-gamma). Cell cultures exposed to both types of IFNs displayed a reduced susceptibility to the cytotoxic activity of human PBL (NK activity). While this effect occurred preferentially at high doses of IFN-alpha, as little as 10 U/ml of IFN-gamma caused a marked decrease in susceptibility to NK-cell-mediated lysis. Using a monoclonal antibody against human beta2-microglobulin (beta2M) a low level of specific binding to K562 cells was detected. The binding increased after treatment with IFN-alpha (1.4-fold) and IFN-gamma (1.7-fold). The expression of transferrin receptors (TR) was not changed significantly. A hybrid cell line between K562 and a Burkitt's lymphoma derived cell line displayed a similar pattern of response to IFN-alpha and IFN gamma as did K562, when effects on NK susceptibility, beta2M expression, and TR expression were studied. The Burkitt's lymphoma line PUT showed no consistent changes in expression of beta2M and TR. These results demonstrate that IFN-gamma is highly efficient in modulating the NK susceptibility, and the expression of beta2M on K562. The presented data do not support a role for expression of TR as the only property that determines the degree of NK susceptibility, since there was no correlation between NK susceptibility and TR expression among the cell lines tested or when IFN-treated and untreated cells were compared. PMID- 2992816 TI - Rhythmic cyclic AMP changes in Chlamydomonas cells synchronized by temperature and light cycles in chemostat culture. AB - Division in chemostat cultures of Chlamydomonas reinhardi was synchronized by daily cycles of both temperature and light. In response to 8 h cool (20 degrees C) - dim light/16 h warm (28 degrees C) - bright light cycles and continuous dilution that removed half the cells daily, the cells replicated nuclear DNA late in the warm-bright phase; released an average of two daughter zoospores early in the warm-bright phase of the next cycle. In these synchronized green algal cultures all ATP and most cAMP were intracellular; most cGMP (approximately 91%) was extracellular. Early in the warm-bright phase ATP doubled after which cAMP level fell 25%. During DNA replication cAMP level doubled, rising to a modest peak reached as replication ended. PMID- 2992817 TI - Membrane phosphatase activities in isolated and cultured adult rat hepatocytes. AB - Membrane bound phosphohydrolysing enzymes, such as Na-K-ATPase, Mg-ATPase, ALPase and G-6-Pase were assayed in intact liver, in freshly isolated cells and in cultured hepatocytes to evaluate the effects of the isolation procedure and culture on these enzyme activities. Na-K-ATPase and Mg-ATPase are significantly reduced following cell dispersion while ALPase and G-6-Pase are nearly unaffected. During culture, Na-K-ATPase is restored to the "in vivo" level within the first two days, but rapidly declines in the following days. The Na dependent, energy requiring AIB uptake shows a similar pattern; Mg-ATPase is practically unmodified. A significant increase in ALPase activity and a net decrease of G-6 Pase activity, as a function of the culture time has been observed. PMID- 2992819 TI - [Preparation of 4-phenoxy and 4-phenylthio derivatives of 1H-pyrazolo[3,4-b] quinoline with an antiviral effect]. PMID- 2992818 TI - Removal of large lipid droplets from cytochalasin B-treated chondrocytes by high speed centrifugation. AB - Chondrocytes isolated from auricular cartilage of 10-week-old rabbits were exposed to cytochalasin B and centrifuged in Percoll solution at 50.000 g for 1 h. This treatment removed the large fat droplets present in the majority of chondrocytes without impairing their viability. It appears that after destruction of actin filaments by cytochalasin, large fat droplets are extruded from the cells during high speed centrifugation due to the difference in the specific weight of fat and chondrocyte cytoplasm. Conceivably, on the same principle other materials could also be eliminated from the cells. PMID- 2992820 TI - [Diffuse glioblastoma of the spinal cord]. PMID- 2992821 TI - [Congenital generalized form of cytomegalovirus inclusion disease in neonates]. PMID- 2992822 TI - Non-mutagenicity of YTR 830 H, a novel beta-lactamase inhibitor. AB - The Ames test indicated that a novel beta-lactamase inhibitor YTR 830 H, belonging to penicillanic acid sulfone was non-mutagenic for any of the tester strains of Salmonella typhimurium TA 98, TA 100 and TA 102. The test was performed both in the presence and absence of S9 activation mixtures which were prepared from the livers of Aroclor 1254-induced Syrian hamsters. PMID- 2992824 TI - Are free radicals involved in tumor promotion? AB - Previously it was shown that lipophilic analogs of a free-radical scavenger, 2(3) tert-butyl-4-hydroxyanisole (BHA), inhibit ornithine decarboxylase (ODC) activity which is induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in mouse epidermis. With regard to this antitumor-promoting effect, eight analogs of BHA (2- and 3-BHA, 2-t-butyl-1, 4-dimethoxybenzene methyl-BHA), t-butylhydroquinone (t-BHQ), p-hydroquinone (HQ), 4-hydroxyanisole, phenol and 2-t-butylphenol) are evaluated herein for their antioxidant capacities for scavenging superoxide anions (O-2), of inhibiting lipid peroxidation and of inhibiting chemiluminescence (CL) in TPA-activated polymorphonuclear leukocytes (PMNs), an event associated with oxy-radical production. None of the analogs reacted with O 2, while 2- and 3-BHA suppressed the formation of O-2 by TPA-activated PMNs. T BHQ underwent autoxidation in aqueous solution, reducing molecular oxygen and increasing the levels of O-2 that were formed chemically, enzymatically and cellularly. However, all of the phenolic antioxidant analogs of BHA inhibited TPA stimulated CL in PMNs and ascorbate-initiated lipid peroxidation, while methyl BHA (a non-antioxidant analog) was inactive. The inhibitory activities of these analogs for lipid peroxidation were related to both their lipophilic and antioxidant properties and corresponded favorably with their inhibitory activities for TPA-induced ODC activities in mouse epidermis. On the other hand, inhibition of the CL response by these antioxidants was independent of their lipophilicity and compared less favorably with their capacities to antagonize phorbol ester-induced ODC activity. These results imply that lipophilic BHA analogs inhibit TPA-induced ODC activity by scavenging free radicals other than O 2. Furthermore, the fact that t-BHQ was the most potent inhibitor of CL, lipid peroxidation and ODC activity and simultaneously reduced molecular oxygen, suggests the possibility that O-2 may act as a precursor to the formation of free radicals which are reactive with t-BHQ and more directly involved in the process of tumor promotion. PMID- 2992823 TI - Studies of self-inactivation of bovine seminal fluid NAD glycohydrolase. AB - Bovine seminal fluid NAD glycohydrolase (NADase) was observed to be rapidly inactivated during catalytic hydrolysis of the substrate NAD. The first-order rate constant for the self-inactivation process was independent of enzyme concentration. The enzyme self-inactivation was a turnover-related process and the number of moles of NAD hydrolyzed required for inactivation was proportional to the enzyme concentration. A number of dinucleotides serving as substrates for the enzyme also promoted self-inactivation. The self-inactivation was an irreversible process having a different rate-limiting step from NAD hydrolysis and was not related to the reversible binding of products and substrate competitive inhibitors. Modification of arginine residues of the enzyme resulted in the loss of NAD hydrolase activity with no differential effect on the self inactivation process. PMID- 2992825 TI - Poly(adenosine diphosphate-ribosylation) of nuclear matrix proteins in alkylating agent resistant and sensitive cell lines. AB - Using Walker 256 breast carcinoma cell lines either with or without acquired resistance to alkylating agents, the structural framework proteins of the nucleus, the nuclear matrix proteins, were found to be effective acceptors for poly(ADP-ribose). Incubation of isolated nuclei with nicotinamide adenine [32P] dinucleotide ([32P] NAD), followed by the isolation of the nuclear matrix, demonstrated that two polypeptides of approximate molecular weight (Mr) 105 000 and 116 000 were extensively poly(ADP-ribosylated). By an in vitro [32P] NAD assay, the nuclear matrix fraction was found to maintain approx. 15% of the total nuclear matrix activity of poly(ADP-ribose) polymerase. Confirmation that the trichloroacetic acid (TCA) precipitable material represented ADP-ribose units was achieved by enzymatic digestion of the nuclear matrix preparation with snake venom phosphodiesterase (SVP). Within 15 min, greater than 85% of the 32P label was digested by SVP and the final digestion products were found to be phosphoribosyl-AMP (PR-AMP) and adenosine 5'-monophosphate (5'-AMP) by thin layer chromatographic analysis. The average polymer chain length was estimated to be 6 7 ADP-ribose units. Because poly(ADP-ribose) polymerase has a putative role in DNA repair, a comparison of the nuclear matrix fractions from Walker resistant and sensitive tumor cell lines was made. In both cell lines, the quantitative and qualitative patterns of the nuclear matrix associated poly(ADP-ribosylation) were similar. PMID- 2992826 TI - [Endotoxemia in liver diseases and its treatment with combined traditional Chinese and Western medicine]. PMID- 2992827 TI - Gestational trophoblastic tumors. AB - Although virtually 100% of women who develop gestational trophoblastic tumors enter a long-term complete remission, there are many aspects of trophoblastic disease that arouse interest. Epidemiological studies have shown a large geographical variation in the percentage of conceptions that result in a hydatidiform mole and have stimulated studies on the immunological differences of the low and high risk populations. Chromosomal analysis is now complementing the pathological differentiation between complete and partial moles. There is still debate as to which factors are positively associated with the progression of a hydatidiform mole through invasive mole to choriocarcinoma. There are also considerable differences in the proportion of molar patients receiving chemotherapy in different centers. In addition to these topics, this article will review several recently introduced treatment regimens which show improved results with reduced toxicity. PMID- 2992828 TI - [Comparative study of the prevalence of Herpesviridae in an African population and a European population]. AB - We have looked for long-term antibodies to Herpesviridae (HSV, VZV, CMV, EBV) in an African population (suburban area of Dakar, Senegal, West Africa) and in a European population (urban area of Bordeaux, France) in order to determine the prevalence of these viruses. The studied sera have been dispatched into 5 age groups: 6 months to 4 years, 5 years to 9 years, 10 years to 14 years, 15 years to 44 years and greater than 45 years. We note that primary infection with Herpesviridae occurs sooner in life in Africa and that the prevalence rates of these viruses are higher in Africa with the exception of EBV, the prevalence of which is almost identical within the fifth age-group of the African and European populations. The prevalence rates of each individual virus are in agreement with previous studies except for VZV which we describe as the least prevalent virus of the family. The interest of this work is a simultaneous comparison of the prevalence of all viruses of the family Herpesviridae within two different populations. PMID- 2992829 TI - [Sampling of urban waste waters in Tananarive. Physical, bacteriological and virological analysis]. AB - After collecting some waters of Tananarive and surroundings in 1975, 1976, 1977, we researched at once enteroviruses, salmonellas and pseudomonas. The results indicated the presence of these agents, often concomitant, in the water samples. An epidemiological study of these microorganisms in waters and some diseases at the same time would be interesting... We hope these researches will begin again soon between Strasbourg and Tananarive. PMID- 2992830 TI - [Monkey pox virus infection in humans in the Central African Republic]. AB - A human monkeypox outbreak is reported which occurred in January 1984 in the extreme south-west areas of the Central African Republic. Six persons were found to be affected in a Pygmy camp with an estimated population of 50 residents. In the two affected families, out of 11 members, only unvaccinated children and a 22 year old unvaccinated woman contracted the disease. The disease was of moderate severity in two patients and very mild in the other four. The clinical diagnosis was confirmed by virus isolation from skin lesions of 4 patients and by sero immunologic tests in all of them. The clinically apparent monkeypox case reported in the Central African Republic in 1983, the presently described outbreak, as well as information on the disease obtained from Pygmies and missionary paramedical staff who are in frequent contact with them, suggest that monkeypox is enzootic in the tropical rain forest in the south-west areas of the Central African Republic. PMID- 2992831 TI - [Poliomyelitis in New Caledonia. Virological and serological data. Pre-immune status and post-vaccinal seroconversion. Prophylactic strategy]. AB - Five cases of paralytic poliomyelitis were noticed between August and November 1982 in the French Territory of New-Caledonia. An epidemiological study allowed to have a better apprehension on the coming back of poliovirus in the South Pacific where, for the past ten years, except in Papua New Guinea which remained an endemic zone, not a single case of poliomyelitis has been declared to the World Health Organization. PMID- 2992832 TI - Analysis of decay phase of synaptic currents from hamster submandibular ganglion cells. PMID- 2992833 TI - A study of the effect of tellurium upon dental caries development in rats: the effect of sodium tellurite and dimercapto succinic acid combination on experimental dental caries development. PMID- 2992834 TI - Novobiocin inhibition of DNA excision repair may occur through effects on mitochondrial structure and ATP metabolism, not on repair topoisomerases. AB - Novobiocin inhibits DNA topoisomerases. It also inhibits excision repair of DNA photodamage, blocking both repair synthesis and the earlier step of incision at u.v. damage sites (as measured by the accumulation of DNA strand breaks in u.v. irradiated interphase cells treated with DNA synthesis inhibitors such as hydroxyurea or cytosine arabinoside). It has been supposed, therefore, that novobiocin affects repair by blocking a putative topoisomerase step prior to incision. But we find that novobiocin also has a marked dose- and time-dependent effect on mitochondria: in cells exposed to novobiocin, mitochondria swell and their cristae become disrupted, and the intracellular ATP:ADP ratio is lowered, though the membrane potential is maintained as judged by rhodamine 123 fluorescence. Mitotic cells are more resistant to mitochondrial disruption by novobiocin than are interphase cells. This correlates with a relative resistance of u.v.-irradiated mitotic cells to the inhibition of incision by novobiocin. The chromosomal decondensation that results from the accumulation of DNA breaks due to incision when u.v.-irradiated mitotic cells are treated with hydroxyurea and cytosine arabinoside is largely suppressed by novobiocin. Furthermore, the suppression of induced strand break accumulation is partly due to a suppression by novobiocin of the uptake and phosphorylation of cytosine arabinoside; breaks accumulated in u.v.-irradiated cells in the presence of aphidicolin, an inhibitor of DNA polymerase alpha that does not require phosphorylation, are less novobiocin-sensitive. We conclude that the effects of novobiocin on excision repair are more likely to be due to a non-specific effect on ATP metabolism than to a specific effect on a repair-related topoisomerase. PMID- 2992835 TI - Inhibition of metabolic cooperation by phorbol esters in a cell culture system based on adenosine kinase deficient mutants of V79 cells. AB - In Chinese hamster V79 cells, stable mutants which are greater than 1000-fold resistant to the adenosine analog, tubercidin (Tubr mutants), and which exhibit high degree of cross-resistance to various other adenosine analogs, viz. toyocamycin, formycin A, 6-methylmercaptopurine riboside and 8-azaadenosine, have been isolated. The inability of the mutant cells to phosphorylate [3H]tubercidin and lack of adenosine kinase activity (AK- phenotype) in their cell extracts provide evidence that the mutant cells are unable to convert adenosine analogs into their toxic phosphorylated derivatives. When AK- cells are co-cultured with increasing numbers of parental V79 (AK+) cells in medium containing tubercidin, then due to metabolic cooperation between AK+ and AK- cells, a cell density dependent decline in recovery of the resistant cells is observed. However, diphtheria toxin resistant (Dipr) mutants of V79 cells, which are altered in elongation factor-2, showed no similar cell density effect. Addition of 0.01 microgram/ml of phorbol ester, 12-O-tetradecanoyl-phorbol-13-acetate (TPA) to growth medium in these experiments markedly enhanced the recovery of the Tubr mutants, but it had no effect on the recovery of Dipr mutants, which suggests that TPA was enhancing recovery of AK- mutants by inhibiting metabolic cooperation between AK- and AK+ cells. Maximum effect of TPA on the recovery of AK- mutants (3- to 5-fold enhancement) was observed at a density of 6 X 10(5) V79 cells/60 mm diameter dish and it was independent of the particular adenosine analog that was used as selective agent. Studies with a number of different phorbol derivatives show that only those phorbol esters which show tumor promoting activity in the mouse skin system inhibited metabolic cooperation between AK+/AK- cells in a dose-dependent manner. An excellent correlation was observed in these studies between the relative tumor promoting activity of various phorbol esters, their relative binding affinities to the cell surface receptors, and the concentrations at which they inhibited metabolic cooperation in the AK-/AK+ cell system. The AK-/AK+ cell system thus provides a new system for examining the effect of tumor promoters on metabolic cooperation between cells. PMID- 2992836 TI - Decreased pressor reactivity to angiotensin II in cirrhotic rats. Evidence for a post-receptor defect in angiotensin action. AB - We used a model of cirrhosis in the rat, produced by inhalation of carbon tetrachloride for 6 weeks, to investigate the mechanism of resistance to the pressor effects of angiotensin II. The pressor response to angiotensin II was significantly lower in conscious cirrhotic animals than in controls. On the other hand, cirrhotic animals had normal pressor responses to norepinephrine, indicating that a generalized defect in vascular reactivity does not cause the decreased pressor response to angiotensin II. Enhanced baroreceptor activity was not the cause of the decreased pressor response to angiotensin II, since baroreflex control of heart rate after angiotensin II was similar in cirrhotics and controls. Pretreatment with either the converting enzyme inhibitor captopril to reduce circulating angiotensin II or the prostaglandin synthesis inhibitor meclofenamate failed to normalize the response to angiotensin II. Thus, neither prior occupancy of receptors with endogenous angiotensin II nor the production of vasodilatory prostaglandins was responsible for the decreased angiotensin II response. Studies of angiotensin II binding by mesenteric artery smooth muscle particles showed that, in cirrhotic animals, receptor affinity for angiotensin II, was significantly lower than in controls (kd: cirrhosis 1.11 +/- 0.09 nM, control 0.94 +/- 0.13 nM; P less than 0.02), whereas receptor number was significantly increased (cirrhosis 315 +/- 42 fmol/mg protein, control 277 +/- 43 fmol/mg protein, P less than 0.01). However, total binding of AII by vascular receptors from cirrhotics was no different than in controls, since the decrease in affinity negated the increase in receptor number.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2992837 TI - Phorbol ester- and diacylglycerol-mediated desensitization of cardiac beta adrenergic receptors. AB - There are specific phorbol ester receptors on cardiac myocytes which may be identical with the calcium/phospholipid-dependent protein kinase (protein kinase C). Incubation of enzymatically dissociated rat cardiac myocytes with biologically active phorbol esters (such as 4 beta-phorbol-12, 13-dibutyrate and 12-O-tetradecanoyl phorbol-13-acetate) leads to a time- and concentration dependent loss of beta-adrenergic receptors detectable with the hydrophilic ligand [3H]-CGP-12177. This loss is attributable to a reduction in both maximal beta-receptor numbers and their affinities. The synthetic diacylglycerol, 1-oleyl 2-acetyldiglycerol, which is known to activate protein kinase C, also induces desensitization of beta-receptors. Both phorbol dibutyrate and 1-oleyl-2 acetyldiglycerol have additive effects to isoproterenol, suggesting a separate site of action in promoting beta-receptor desensitization. The effects of phorbol dibutyrate and 1-oleyl-2-acetyldiglycerol are prevented by colchicine (but not its inactive analog, trimethylcolchicinic acid), indicating a microtubule dependence. The loss of membrane-bound beta-receptors after phorbol dibutyrate- or 1-oleyl-2-acetyldiglycerol preincubation is accompanied by an increase in beta receptors associated with a cytosol-derived vesicular fraction devoid of plasma membrane markers, a finding consistent with an internalization process. These results suggest that protein kinase C activation by diacylglycerols derived from receptor-linked phosphoinositide hydrolysis may be a novel mechanism of cardiac beta-receptor desensitization. PMID- 2992838 TI - Comparative systemic and renal effects of dopamine and angiotensin-converting enzyme inhibition with enalaprilat in patients with heart failure. AB - Renal and systemic hemodynamics were measured during titration of dopamine and serially after intravenous administration of enalaprilat in nine patients with chronic severe congestive heart failure. During titration of dopamine, renal blood flow increased by 99%, from 304 +/- 120 to 604 +/- 234 ml/min (p less than .01) at a dose of dopamine of 2.1 micrograms/kg/min, which produced only a 21% increase in cardiac index, from 1.96 +/- 0.36 to 2.38 +/- 0.35 liters/min/m2 (p less than .05). Cardiac index was increased maximally at a dose of 4.0 micrograms/kg/min dopamine; however, renal blood flow was not further augmented. In contrast, after intravenous administration of enalaprilat, peak improvement of renal blood flow and cardiac index occurred concomitantly. Renal blood flow increased by 35%, from 316 +/- 97 to 427 +/- 107 ml/min (p less than .05), and cardiac index increased by 18%, from 1.99 +/- 0.40 to 2.35 +/- 0.40 liters/min/m2 (p less than .05). At similar increases in cardiac index, dopamine produced a greater increase in renal blood flow than enalaprilat: 604 +/- 234 vs 427 +/- 107 ml/min (p less than .05). Mean systemic arterial pressure, however, was greater with dopamine than with enalaprilat (78.1 +/- 16.7 vs 70.2 +/- 17.2 mm Hg; p less than .05) at peak effect. Thus, although both drugs appear to be potent renal vasodilators in patients with severe congestive heart failure, dopamine may be more effective in augmenting renal blood flow. PMID- 2992839 TI - Attenuation of exercise-induced asthma by pretreatment with nedocromil sodium and minocromil. AB - In a group of atopic adult asthmatic patients the effects of two new inhaled antiasthmatic drugs, nedocromil sodium and minocromil were studied in two independent randomized double-blind trials to assess their efficacy in preventing exercise-induced asthma (EIA). Exercise testing consisted of steady state running on an inclined treadmill. Neither drug administered 30 min before exercise significantly altered baseline FEV1. Nedocromil sodium (2 and 4 mg) pre-treatment in nine patients and minocromil (4 mg) in eight patients gave significant protection (P less than 0.001) compared to placebo as assessed by the reduction in the maximum percentage fall in FEV1. There was no significant difference in the inhibitory effect of the two doses of nedocromil sodium. These results indicate that both nedocromil sodium and minocromil can attenuate EIA. PMID- 2992840 TI - Gene probes: application to prenatal and postnatal diagnosis of genetic disease. AB - Gene probes can now be used to detect a variety of mutations that produce single gene disorders. In present clinical practice, restriction endonuclease analysis is used for the prenatal diagnosis of sickle cell anemia, alpha-thalassemia, and beta-thalassemia. Direct detection of the mutation is possible in alpha thalassemia, where a deletion has usually occurred, and in sickle cell anemia, where the mutation alters the recognition sequence of the restriction endonuclease, Mst II. Indirect detection of beta-thalassemia is based on using normal variations in DNA (DNA polymorphisms) to track normal and affected beta globin genes in families. This latter kind of analysis is also useful in detecting the phenylalanine hydroxylase genes affected in phenylketonuria and will often be used in disorders where the mutations are unknown. In cases where the mutation is known, direct analysis by use of oligonucleotide probes is a new and important advance. An example of this type of gene detection in a family with classical hemophilia is presented. In addition, with chorion villus biopsy, detection of these inherited diseases is feasible by the 12th week of pregnancy. PMID- 2992841 TI - The use of DNA probes in studies of human cytomegalovirus. AB - Hybridization assays provide a sensitive and rapid means for studying the molecular biology of viral replication and for identifying viral nucleic acid in biological specimens. Such assays are attractive because the detection of virus does not require intact virions or concomitant viral protein synthesis, both of which may be absent in a latently infected cell or in a virus-associated tumor. For molecular and clinical studies on human cytomegalovirus (HCMV), we have cloned and characterized subgenomic EcoRI fragments representative of the entire genome of HCMV strain AD169. To study the epidemiology of HCMV infections and to identify the presence of HCMV nucleic acid in urine, blood, Kaposi's sarcoma, and other tissues, we have used various hybridization techniques, including DNA dot/slot-blot hybridization, Southern blot hybridization, and in situ cytohybridization. These studies demonstrate how cloned molecular probes can be used to study the molecular biology, pathogenesis, and treatment of viral infections. PMID- 2992843 TI - What is the "correct" activity of angiotensin-converting enzyme? PMID- 2992842 TI - Radiolabeled drugs as probes of central nervous system neurons. AB - Methylspiperone, carfentanil, and suriclone, labeled with 11C, can be used in assessment of dopamine, mu-type opiate receptors, and benzodiazapine receptors in the brains of living human beings. These relatively specific neuronal probes permit measurement of the number and state of occupancy of receptors in the brain in health and disease, and may be useful to assess endogenous neurotransmitter secretion by decreasing competitive binding of the labeled ligand. To date, the most striking finding has been a decrease in the indices of dopamine and serotonin receptor densities as a function of age in normal men and women. A possible relationship between this decrease and vitamin B6 deficiency is currently under study. PMID- 2992844 TI - Chromatographic characterisation of neurotensin-like immunoreactivity in plasma and tissue extracts from hepatoma patients. AB - To determine the nature of the neurotensin-like immunoreactivity produced by some cases of primary liver carcinoma, plasma and tumour neurotensin-like immunoreactivities were characterised in patients with neurotensin-producing hepatoma by chromatographic analysis in conjunction with radioimmunoassay with C- and N-terminally directed antisera. Gel filtration revealed that the majority of neurotensin-like immunoreactivity extracted from the tumour was co-eluted with synthetic neurotensin, whereas a substantial amount of the plasma immunoreactive neurotensin emerged at the void volume. Tumour neurotensin-like immunoreactivity showed a degree of heterogeneity on gel filtration and high pressure liquid chromatography that contrasted with neurotensin-like immunoreactivity extracted from ileal mucosa, suggesting a different post-translational processing of neurotensin within tumour tissue. The ratio of neurotensin 1-8 to neurotensin 1 13 in the patients' plasma suggested that neurotensin was metabolised in a manner similar to that in healthy subjects. The clinical and biological significance of the ectopic production of neurotensin in some hepatomas remains unclear. PMID- 2992845 TI - Estimation of 1,25 dihydroxyvitamin D by cytoreceptor and competitive protein binding assays without high pressure liquid chromatography. AB - Using two different cultured rat osteosarcoma cell lines (UMR 106 and ROS 17/2.8) we have investigated the recently described cytoreceptor assay for 1,25 dihydroxyvitamin D (1,25-(OH)2D). The assay method is relatively simple and sensitive to 2.4 fmole per tube. Using either cell line, assay of serum samples, whose only preparation consisted of extraction and purification on a disposable diatomaceous earth column, produced variable values for serum 1,25-(OH)2D. Additional purification, using a disposable silicic acid minicolumn to remove other vitamin D metabolites resulted in consistent values and additional HPLC resulted in no further decrease in the values obtained. Our results show that a single two stage non-HPLC column can purify serum samples for assay in the cytoreceptor assay. The method is also applicable to the competitive protein binding assay employing calf thymus cytosol and the correlation between values obtained by both methods is highly significant. It is a sensitive, simple, and accurate method with technical advantages which allow greater sample throughput than other 1,25-(OH)2D assays. PMID- 2992847 TI - Effect of nivazol on ACTH secretion by human pituitary corticotrophic tumours in cell culture. AB - The effects of nivazol, a novel steroid which lacks glucocorticoid activity, on ACTH secretion by cell cultures of human pituitary corticotrophic tumours has been investigated. Nivazol (0.002-20 mumol/l) inhibited ACTH secretion by 50-80%, after 4 and 24 h of incubation. (Trilostane (0.3-30 mumol/l) did not affect basal or stimulated ACTH secretion.) The potency of nivazol was comparable with that of hydrocortisone, and less than dexamethasone. The stimulatory effects of oCRF and AVP were reduced or completely blocked by nivazol in a similar manner to that previously described for hydrocortisone. It is concluded that nivazol may be of benefit in the direct treatment of Cushing's disease, particularly since it lacks glucocorticoid activity. PMID- 2992846 TI - Augmentation of the response to CRF in man: relative contributions of endogenous angiotensin and vasopressin. AB - The plasma ACTH response to corticotrophin releasing factor (CRF) was studied in seven normal individuals. Five subjects were restudied following 4 d of salt restriction which resulted in raised endogenous plasma angiotensin II secretion. In a third experiment six subjects were given CRF following pre-infusion of hypertonic saline which significantly increased endogenous plasma vasopressin (AVP) levels. We were unable to demonstrate that high endogenous plasma AII levels were associated with a significant change in the plasma ACTH or cortisol responses to CRF. However there was an almost three-fold increase in the ACTH response when endogenous plasma AVP was elevated by hypertonic saline. It is concluded that AVP is likely to be of physiological importance in potentiating the ACTH response to CRF and that AVP and CRF together may provide a better test of pituitary ACTH secretion than either peptide alone. PMID- 2992848 TI - Corticotrophin, cortisol, prolactin and growth hormone responses to insulin induced hypoglycaemia in normal subjects given sodium valproate. AB - Plasma corticotrophin (ACTH), cortisol, prolactin and growth hormone (GH) responses to insulin-induced hypoglycaemia were measured in normal healthy subjects of both sexes before and after three weeks' treatment with sodium valproate (Epilim, 200 mg three times a day). The drug had no effect on fasting plasma glucose levels, or the extent of hypoglycaemia induced by insulin (0.15 U/kg). There was no significant difference between pre- and post-treatment values for basal or stress-induced concentrations of ACTH and cortisol (n = 12), prolactin (n = 7) or GH (n = 9). The results suggest that treatment of normal subjects with sodium valproate has no effect on the response of the hypothalamo pituitary-adrenocortical axis to hypoglycaemia, which is in contrast to its inhibitory effects on ACTH secretion in patients suffering from Nelson's syndrome. This implies that in the disease state, there may be a unique sensitivity to GABA-ergic manipulation. PMID- 2992849 TI - Adenoid cystic carcinoma of the upper digestive and respiratory tracts. A retrospective study of 59 cases. AB - Fifty-nine cases of adenoid cystic carcinoma (ACC) in the head and neck were reviewed and the treatment and follow-up results from 1950 to 1983 are reported. Nearly all patients were treated with combined therapy, i.e. surgery and radiotherapy, and the actuarial survival rate was 68% after 5 years, 45% after 10 years, and 30% after 15 years. Modern radiotherapy seems a very valuable means of treatment, not only as a palliative measure. PMID- 2992850 TI - The portal system. PMID- 2992851 TI - The liver. PMID- 2992852 TI - Genetic counseling and genetic heterogeneity in the thalassemias. AB - In this study, we have compared the hemoglobin A2 levels (Hb A2) of alpha thalassemia carriers (-alpha/-alpha and -alpha/alpha alpha genotypes) with those of double heterozygotes for delta+ and beta thalassemia genes, who were identified by family studies and polymorphic restriction site analysis within the beta-globin gene cluster. We found that double heterozygotes for the delta+ and beta thalassemia have significantly (p less than 0.001) higher Hb A2 levels as compared with carriers of alpha-thalassemia. This finding has practical implications in the genetic counseling of subjects with a thalassemia-like phenotype associated with normal or borderline Hb A2 levels. PMID- 2992853 TI - Investigations into the nature of alpha 2-adrenoceptors in rat tail arteries. AB - In rat isolated perfused tail arteries, dose-response curves were established for the vasopressor effects of phenylephrine (alpha 1-adrenoceptor agonist), clonidine (alpha 1- and alpha 2-adrenoceptor agonist), clonidine in the presence of 10(-7) mol/l prazosin (alpha 2-agonist), and BHT-920 (alpha 2-agonist). The ED50 values were: phenylephrine 1.85 X 10(-10) mol; clonidine 6.3 X 10(-10) mol; clonidine + prazosin 3.2 X 10(-6) mol; BHT-920 6.1 X 10(-6) mol. The arterial reactivity to BHT-920 was stable only after 4-5 h of perfusion. Responses to BHT 920 were not antagonized by yohimbine (alpha 2-adrenoceptor antagonist) but were antagonized by low concentrations of prazosin (alpha 1-adrenoceptor antagonist). These data constitute conflicting evidence regarding the existence of alpha 2 adrenoceptors in rat tail arteries. The data are consistent with the proposal that there are two recognition sites on alpha 1-adrenoceptors; phenylephrine and BHT-920 may stimulate different sites on alpha 1-adrenoceptors. PMID- 2992854 TI - Potentiation of ACTH hypertension in man with salt loading. AB - ACTH 1 mg/day for 5 days raises systolic blood pressure (SBP) in normotensive and hypertensive subjects on a fixed electrolyte intake of 100 mmol/day sodium (Na) and potassium (K) (Whitworth et al. 1983). The present study examined the effect of Na intake in the high normal range on the haemodynamic and metabolic responses to ACTH. ACTH administration on a 200-300 mmol Na intake increased SBP by 35 mmHg (s.e.m. = 11, n = 5) compared with the 22 mmHg (s.e.m. = 4, n = 12) rise seen on a 100 mmol Na intake in our previous study. These studies suggest that the effects of adrenocortical steroids on blood pressure in man may be magnified by increasing dietary Na intake. PMID- 2992855 TI - Hypertension corrected and aldosterone responsiveness to renin-angiotensin restored by long-term dexamethasone in glucocorticoid-suppressible hyperaldosteronism. AB - Two males with glucocorticoid-suppressible hyperaldosteronism had hyperaldosteronism, hypertension and hypokalaemia corrected by continuous administration of physiological doses of dexamethasone for more than a year. During long-term dexamethasone treatment: (a) Plasma renin activity increased from subnormal to high normal levels, with normal posture-mediated increases; (b) Plasma aldosterone became responsive to angiotensin infusion, a new observation; (c) A fall in plasma aldosterone between 0800 h (recumbent) and 1000 h (upright) was replaced by a rise; (d) Plasma aldosterone became suppressible with salt loading. These findings are consistent with a shift to more normal control of aldosterone by renin-angiotensin, once abnormal responsiveness to ACTH has been nullified. PMID- 2992856 TI - Cellular targets of antilymphocyte antibodies in AIDS and LAS. AB - Many patients with acquired immune deficiency syndrome (AIDS) and lymphadenopathy syndrome (LAS) have serum antilymphocyte antibodies. The targets of these antibodies are neither sex nor HLA directed. There is disagreement about the cell subset at risk. We examined the cellular specificity of antilymphocyte antibodies from AIDS and LAS patients by microcytotoxicity on positively selected lymphocytes, by using double-labeling immunofluorescence, and by using well characterized continuous cell lines as targets. We find that most AIDS/LAS patients have antibodies to T and B cells, that immunofluorescence is somewhat more sensitive than microcytotoxicity, and that the antibodies are directed to both T4+ and T4- lymphoid cells. PMID- 2992857 TI - Characterization of Epstein-Barr virus-specific T-cell hybridomas derived from infectious mononucleosis. AB - Somatic cell hybridization was utilized to produce hybrids with surface receptors that would pertain directly to those expressed in vivo during Epstein-Barr virus (EBV) infection. Lymphocytes were obtained during acute infectious mononucleosis (IM) and fused to a double mutant of the JM human T-lymphoma cell lines. Hybrid cells that reacted with autologous EBV-infected lymphoblasts were detected by the release of Interleukin-2 into the culture medium. Reactive hybridomas also released IL-2 following coculture with allogeneic EBV-infected cells when those cells shared HLA-DR antigens of the primary parental cells. In contrast, stimulator cells with no shared HLA-DR or without evidence of EBV infection never induced IL-2 release. These results suggest the existence of a population of T cells that arise during acute IM and could account for the known proliferative phase of the disease. The requirements of IL-2 stimulation are currently under study using this system. PMID- 2992858 TI - Cell-mediated immunity in human herpesvirus infection: analysis by monoclonal antibodies. AB - Peripheral blood lymphocytes (PBL) were obtained from heterosexual patients during the first or recurrent episodes of genital herpes simplex type 2 (HSV-2) infections. These cells were incubated with a panel of lymphocyte-specific monoclonal antibodies and examined by indirect immunofluorescence and flow cytometry. Lymphocyte proliferative responses to inactivated HSV-2 (UV-HSV-2) were also determined on repeat occasions using PBL obtained from patients with first episodes of infection. Flow cytometry analysis indicated that PBL obtained within 1 week after the onset of symptoms of first episode genital herpes exhibited significantly lower OKT 4 helper/OKT 8 suppressor T-cell ratios (1.26 +/- 0.12) than cells from either normal controls (2.44 +/- 0.3, P less than 0.01) or patients with recurrent genital HSV-2 infection (2.58 +/- 0.40, P less than 0.01). This lowered ratio was due to a decrease in OKT 4-positive helper T cells present early during initial infection (30.2 +/- 2.9, P less than 0.01 compared to recurrent disease patients or controls) and was not caused by a concomitant increase in OKT 8-positive T cells. No variation was observed during the course of infection in the proportion of total T cells as determined by EAET rosette formation or reactivity with the T-cell specific monoclonal antibody 9.6. PBL obtained from patients within 1 week after symptom onset exhibited a poor proliferative response to UV-HSV-2. Maximal proliferative responses occurred 6-9 weeks after disease onset and required the presence of OKT 4-positive T lymphocytes. The results of these experiments indicate that at different stages of disease, HSV infection is accompanied by fluctuations in the ratio of helper to suppressor T cells and alterations in antigen-specific lymphocyte proliferation. These findings suggest that variations in lymphocyte proliferative responses during the course of first episode genital herpes infection may reflect disease related variations in either the number or activity of circulating OKT 4 positive T lymphocytes. PMID- 2992859 TI - Lymphocyte subsets and surface molecules in man. PMID- 2992860 TI - Salivary adenoid cystic carcinoma. Features of an intra-abdominal metastasis. AB - An adenoid cystic carcinoma of salivary gland origin metastasized to the abdomen four years after the initial diagnosis. It showed heavy uptake of Ga-67 citrate and hypervascularity. The clinical and pathologic features are discussed. Gallium scan and arteriography (usually external carotid artery injection) may prove helpful in the early diagnosis and management in the various phases of the disease. PMID- 2992861 TI - Single photon emission computed tomography and renal pseudotumor. AB - Single photon emission computed tomography (SPECT), recently introduced into clinical nuclear medicine, potentially offers improved spatial resolution compared to planar imaging by eliminating activity from surrounding structures, and permitting three-dimensional reconstructions. Its clinical applications to date have been limited. A case where SPECT with Tc-99m dimercaptosuccinic acid (DMSA) better demonstrated a renal pseudotumor (a benign column of Bertin) compared to planar imaging is reported. PMID- 2992862 TI - False-positive technetium-99m DMSA renal imaging in two cases of malrotated kidney. AB - Technetium-99m dimercaptosuccinic acid (Tc-99m DMSA) has been used as a renal imaging agent of choice. A few false-positive cases of Tc-99m DMSA imaging have been reported. The authors have experienced false-positive Tc-99m DMSA imagings in two cases of a malrotated kidney, and this finding previously has not been reported in the literature. The possibility of its nuclear imaging diagnosis with Tc-99m DTPA is also discussed. PMID- 2992863 TI - Neurofibromatosis and malignancy. AB - Neural crest and nonneural crest tumors occur frequently in neurofibromatosis (NF). We report one case of NF and recurrent malignant fibrous histiocytoma, a tumor that is uncommon in childhood, and another case of the concomitant occurrence of NF, hemophilia B, and a paratesticular rhabdomyosarcoma. PMID- 2992864 TI - Primitive neuroectodermal tumor presenting as meningitis. PMID- 2992865 TI - [Biochemical and clinical principles in the therapy of alcoholism]. PMID- 2992866 TI - Evaluation of CMV total antibody EIA. An enzyme immunoassay for detection of antibodies to cytomegalovirus. AB - The performance of an indirect enzyme-immunoassay (CMV Total Antibody EIA, Abbott Laboratories) for the detection of the presence or absence of antibodies to cytomegalovirus (CMV) was evaluated by investigators at five clinical sites. The interassay and intraassay coefficient of variation data showed that the EIA could be performed in a highly reproducible manner at all sites. The investigators tested a total of 2314 clinical samples for antibody to CMV by the CMV Total Antibody EIA and by an indirect hemagglutination assay (IHA). The overall results demonstrated that compared with IHA the sensitivity of the EIA was 97.9% (1096/1120) and the specificity was 93.6% (1096/1171). The clinical samples tested included sera, plasma, and whole blood. These results show a good agreement between the CMV Total Antibody EIA and the IHA. This EIA is a simple, objective, and reproducible method for the detection of antibodies to CMV in sera, plasma, or whole blood samples, and the total procedure is adaptable for batching and automation and takes only 3 hr of elapsed time. PMID- 2992868 TI - Cytomegalovirus antibody detection by three commercially available assays and complement fixation. AB - Four methods (complement fixation, latex agglutination, qualitative EIA, and quantitative EIA) for detecting antibody to cytomegalovirus (CMV) were compared by testing 103 sera. When ranked according to accuracy, sensitivity, and specificity, the complement fixation test was third, fourth, and first; the latex agglutination test was first, second, and third; the qualitative EIA was fourth, first, and fourth, whereas the quantitative EIA was second, third, and first, respectively. In addition, the complement fixation, latex agglutination, and quantitative EIA systems each satisfactorily detected significant antibody rises in paired sera. PMID- 2992867 TI - A simple method to detect intrathecal production of specific antimeasles antibodies in cerebrospinal fluid during subacute sclerosing panencephalitis. AB - The intrathecal production of antimeasles antibodies was studied using the enzyme linked immunosorbent assay in eight specimens of serum and cerebrospinal fluid (CSF) from patients with clinical signs of subacute sclerosing panencephalitis (SSPE). The test was performed using a 1:5 dilution of CSF and a 1:2000 dilution of serum (ratio 1:400) in order to nullify the physiological gradient of immunoglobulins across the blood brain barrier (BBB). This procedure allowed a rapid and accurate assessment of the synthesis of specific immunoglobulins in the CSF and a good evaluation of the permeability of the BBB. A diagnosis of SSPE was provided in five out of eight patients with clinical signs of the disease. Clinical follow-up confirmed the diagnosis of SSPE in the group of patients with clear evidence of intrathecal synthesis of antimeasles antibodies. PMID- 2992869 TI - Dietary intake in a group of institutionalised elderly and the effect of a fibre supplementation programme on nutrient intake and weight gain. PMID- 2992870 TI - Isolation and characterization of calmodulin from a molluscan smooth muscle. AB - Calmodulin was purified from the anterior byssal retractor muscle (ABRM) of a mollusc Mytilus edulis. Ca2+-induced conformational changes in the ABRM calmodulin could be demonstrated by polyacrylamide gel electrophoresis, by u.v. absorption spectrum and by circular dichroic spectrum. The amino acid composition of the ABRM calmodulin closely resembled that of other invertebrate calmodulins. The ABRM calmodulin was less effective in activating rat brain phosphodiesterase than vertebrate calmodulins. PMID- 2992871 TI - Purification and properties of carp (Cyprinus carpio) muscle calpain II (high Ca2+-requiring form of calpain). AB - Calpain (Ca2+-dependent cysteine proteinase) was purified to apparent homogeneity from carp muscle by the method of DEAE-cellulose, hydroxylapatite and Ultrogel AcA 34 column chromatographies. The purified enzyme is classified as calpain II (high-Ca2+-requiring form of calpain) from the effects of Ca2+ concentration, pH and the antibiotics on the activity. Carp muscle calpain II was inhibited by rat liver calpastatin, the specific inhibitor for calpain. It is probable that the calpain-calpastatin system may play a biologically fundamental and common role in various cells, since the inhibitory effect of calpastatin on calpain from different tissues of different species is well conserved. PMID- 2992872 TI - Mutual inhibitory effect of calpastatins on calpains from carp muscle, carp erythrocytes and rat liver. AB - The inhibitory effect of calpastatin (specific inhibitor for calpain) on calpain (Ca2+-dependent cysteine proteinase, EC 3.4.22.17) was examined using carp muscle, carp erythrocytes and rat liver preparations. A mutual inhibitory effect between calpains and calpastatins from different tissues and species was observed. The conservation of the inhibitory effect of calpastatin on calpain among vertebrates suggests that the calpain-calpastatin system may play a biologically fundamental and common role in various cells. PMID- 2992873 TI - Detection and some properties of calpain II (high-Ca2+-requiring form of calpain) in carp (Cyprinus carpio) erythrocytes. AB - In order to examine the existence of calpain I, a low (micromolar)-Ca2+-requiring form of calpain, in fish tissues, carp erythrocytes were chosen as the experimental material, since only calpain I is known to exist in mammalian erythrocytes. By DEAE-cellulose chromatography, calpain and calpastatin (specific inhibitor for calpain) were separated from carp erythrocyte hemolysate. Carp erythrocyte calpain is classified as calpain II, a high (millimolar)-Ca2+ requiring form of calpain, from the result of Ca2+-requirement for the activity. PMID- 2992874 TI - The p-nitrophenyl phosphatase activity of ubiquitin from bovine erythrocytes. AB - Ubiquitin, a unique protein with esterase and carbonic anhydrase activity, has been found to have also a p-nitrophenyl phosphatase activity. This phosphomonoesterase activity of ubiquitin has an acidic pH optimum; its true substrate appears to be the phosphomonoanion, with a Km of 1.8 X 10(-3) M. It is competitively inhibited by the typical acid phosphatase inhibitors, arsenate (Ki = 1.3 X 10(-3) M), molybdate (Ki = 1.2 X 10(-6) M), and phosphate (Ki = 1.4 X 10( 3) M). These inhibitors have no effect on the CO2 hydration and p-nitrophenyl acetate esterase activities of the ubiquitin. Acetazolamide slightly inhibited the p-nitrophenyl phosphatase activity. PMID- 2992875 TI - Adenylate energy charge, arginine phosphate and ATPase activity in juvenile Homarus americanus during the molt cycle. AB - Neither gill nor hepatopancreas exhibited significant differences in Na+, K+ ATPase activity with molt stage. Hepatopancreatic residual ATPase activity was significantly higher (F = 6.273) in post-molt animals; while gill residual ATPase activity exhibited no significant differences. Muscle AEC did not change with molt stage, but levels of ATP (F = 8.050) and ADP (F = 4.130) were significantly higher in premolt (D3 pleopod stage 5.0-5.5) animals; while levels of arginine phosphate (F = 6.981) were significantly higher in post-molt animals. Arginine phosphate/ATP and ATP/ADP ratios were highest in post-molt animals, but were not statistically significant. Although not significant, changes in Na+, K+-ATPase activity and AEC did suggest alterations in: enzyme activity that correlate with known osmotic compensations occurring during the water uptake and hardening/mineralization processes; and energy metabolism which occur during the molt cycle, respectively. PMID- 2992876 TI - Comparison of enzyme activities on glycogen metabolism in rabbit slow and fast muscles. AB - Activities of glycogen synthase (total) and branching enzyme in slow (soleus) muscle are higher than those in fast (vastus lateralis) muscle, while those of phosphorylase kinase (total), phosphorylase (total) and debranching enzyme are reversed. The active form ratio of glycogen synthase is higher in fast muscle, while those of phosphorylase kinase and phosphorylase are higher in slow muscle. Activities of cAMP-dependent protein kinase and protein phosphatase in slow muscle are higher than those in fast muscle. These results suggest that glycogen metabolizing enzymes in slow muscle, distinct from those in fast muscle, are regulated more strongly by cAMP-dependent protein kinase rather than by protein phosphatase. PMID- 2992877 TI - Metabolism of glucose 1,6-P2. I. Enzymes involved in the synthesis of glucose 1,6 P2 in pig tissues. AB - Pig tissues show four enzymatic activities of glucose 1,6-P2 synthesis: (A) 2 [glucose 1-P]----glucose 1,6-P2 + glucose; (B) glucose 1-P + ATP----glucose 1,6 P2 + ADP; (C) glucose 1-P + fructose 1,6-P2----glucose 1,6-P2 + fructose 6-P; (D) glucose 1-P + glycerate 1,3-P2----glucose 1,6-P2 + glycerate 3-P. Brain is the tissue with highest capability of glucose 1,6-P2 synthesis. With the exception of skeletal muscle, activity "D" represents the highest activity of glucose 1,6-P2 synthesis. In muscle, activity "B" is the major activity. The existence of a specific glucose 1,6-P2 synthase which catalyzes reaction "D" is confirmed. Two peaks of such an enzyme are isolated by ion-exchange chromatography. There is an enzyme which specifically catalyzes reaction "C", not previously described. There is a glucose 1-P kinase not identical to phosphofructokinase. PMID- 2992879 TI - Blocking (suppressor) factors, immune complexes, and extracorporeal immunoadsorption in tumor immunity. PMID- 2992878 TI - Metabolic investigations of fibroblasts from horses, Equus caballus, with hereditary severe combined immunodeficiency. AB - In an attempt to determine the metabolic defect causing severe combined immunodeficiency (SCID) in horses in which altered purine metabolism has been observed, various parameters of purine and pyrimidine metabolism were evaluated. The activities of nine purine enzymes (adenosine kinase, purine nucleoside phosphorylase, deoxyadenosine kinase, deoxycytidine kinase, 5'-nucleotidase, AMP deaminase, hypoxanthine-guanine phosphoribosyl transferase, and adenine phosphoribosyl transferase were measured in fibroblasts. All activities determined for SCID horses were normal. Uptake of 10 microM adenosine or 2' deoxyadenosine (a growth inhibitory concentration for SCID fibroblasts) by SCID fibroblasts was identical to that found for normal fibroblasts in the presence of both 1 and 50 microM phosphate. The Km determined for the transport of both adenosine and 2'-deoxyadenosine was 35 microM. In the presence of p nitrobenzylthioguanosine (a nucleoside transport inhibitor), 2'-deoxyadenosine uptake was inhibited to the same extent in all fibroblast lines tested. To determine if the last step in pyrimidine biosynthesis might be altered in SCID fibroblasts, UMP synthase activities were evaluated but found to be normal (0.5 nmol UMP formed/min/mg protein). PMID- 2992880 TI - The zoapatle. VI. Revisited. AB - A review of new Zoapatle publications was made, including five chemical compounds characterized recently. Relative potency of Kaurenoic acid, Kauradienoic acid (its mixture), Zoapatanol and Montanol was estimated in an in vitro guinea pig uterine assay. Kaurene compounds were several times more potent than Montanol and Zoapatanol as uteroactive substances. Zoapatle aqueous crude extract made from Montanoa frutescens had a strong in vitro uterine potency and unique in vivo characteristics in several biological systems, described in detail in the following five publications. PMID- 2992881 TI - Bacterial chemotaxis: biochemistry of behavior in a single cell. AB - Bacterial chemotaxis is a primitive behavioral system that shows great promise for being amenable to a description of its molecular mechanism. In Gram-negatives like Escherichia coli, addition of amino acid attractant begins a series of events, starting with binding to certain intrinsic membrane proteins, the MCPs, and ending with a period of smooth swimming. Immediately, methyl-esterification of these MCPs begins and continues during this period. By contrast in the Gram positive Bacillus subtilis, demethylation of MCPs occurs during the same period. At least two other mechanisms for mediating chemotaxis toward the attractants oxygen and phosphotransferase sugars exist in E. coli, and in these, changes in methylation of MCPs plays no role. Moreover, chemotaxis away from many repellents by B. subtilis appears to involve different mechanisms. Many of the repellents include drugs and toxicants, many of them man-made, so that chemoreceptors could not have specifically evolved; yet the bacteria are often exquisitely sensitive to them. Indeed, the B. subtilis membrane seems to act like a generalized antenna for noxious membrane-active substances. PMID- 2992882 TI - Cryolumpectomy for breast cancer: an experimental study. AB - The effects of cryosurgical procedures and surgical excision in preventing the local recurrence of mammary adenocarcinoma were studied in BALB/cfC3H mice carrying syngeneic, virus-induced mammary adenocarcinomas transplanted into the fourth mammary fat pad. In this report we present evidence demonstrating that cryosurgical procedures involving multiple freeze-thaw cycles followed by tumor excision markedly reduce the local recurrence rate of mouse mammary cancer. Surgical resection without cryotreatment resulted in an 80% local recurrence rate; in contrast, cryotreatment consisting of three freeze-thaw cycles before excision prevented local tumor recurrence in 70% of the animals. The use of cryotherapy and local excision (cryolumpectomy) in the treatment of human breast cancer is discussed. PMID- 2992883 TI - Role of lymphocyte and antibody in the pathogenesis of immune-mediated herpetic uveitis. AB - We investigated the role of various immune components in the pathogenesis of immune-mediated experimental herpetic uveitis. Inbred III/J strain of rabbits were sensitized with an intravitreal injection of 10(3) PFU of type 1 herpes simplex virus (HSV), and sensitized cervical lymph node (LN) cells were obtained on postinfection day 12. Intravitreal injection to the normal III/J rabbit eye of HSV antigen with either sensitized LN cells or anti-HSV serum failed to induce uveitis, whereas intravitreal injection of HSV-antigen with both sensitized LN cells and anti-HSV serum produced severe uveitis within six hours. The combination of sensitized LN cells, HSV-antigen and normal rabbit serum, or that of normal LN cells, HSV antigen and anti-HSV serum, did not induce uveitis. Further studies using B lymphocyte and T lymphocyte fractions from sensitized LN showed that only the combination of sensitized T lymphocytes, HSV antigen and anti-HSV serum regularly produced uveitis following intravitreal injection. These results indicate that the interaction of HSV antigen, sensitized T lymphocytes and anti-HSV antibody may play a role in the pathogenesis of immune-mediated herpetic uveitis. PMID- 2992884 TI - Epithelial and endothelial effects from the extended wear of contact lenses. PMID- 2992885 TI - Metabolism of phosphoinositides and inositol polyphosphates in rabbit corneal epithelium. AB - The synthesis of phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP) and phosphatidylinositol-4,5-bisphosphate (PIP2) was studied in the rabbit cornea using 32P and [3H]myo-inositol as precursors. Also, the formation of water soluble products of the degradation of inositol lipids, inositol-1-phosphate, 1,4-bisphosphate and -1,4,5-trisphosphate, was shown. Corneal epithelium displayed the most active inositol lipid metabolism and endothelium the least when the cornea layers were separately incubated (stroma had intermediate values). In corneal epithelium incubated with [3H]myo-inositol, tritiated glycerophosphorylinositol and water-soluble inositol phosphates were formed. 10 mM LiCl promotes an increase in the inositol phosphates consistent with an inhibitory effect on inositol phosphatase. Cell-free preparations of epithelium incubated with [gamma-32P]ATP detected the presence of diacylglycerol kinase, PI kinase and PIP kinase. Endogenous PI was rapidly phosphorylated to PIP within 1 min of incubation, whereas PIP was phosphorylated more slowly. In conclusion, the components of the inositol lipid cycle are present in the cornea, particularly in the epithelium. It is proposed that the control of these pathways may be involved in the transduction of cell signals through the plasma membrane, intracellular calcium ionization and epithelial cell proliferation and differentiation, particularly in wound healing. PMID- 2992886 TI - Inhibition of collagenase activity by extracts of bovine ocular tissues. AB - Bovine eyes were dissected and separate pools of lens, lens capsule, cornea and vitreous were extracted in guanidine, subjected to ultrafiltration, and examined for their effects on collagenolytic activity. Although lens extract was not inhibitory, the cornea and vitreous both contained inhibitors of collagenase. More inhibition was present in the filtrate of the vitreous than in the retentate, whereas the total amount of inhibition in the cornea was distributed almost equally between the two fractions. The inhibition observed was dose dependent. The partially purified inhibitors from cornea and vitreous blocked the activity of human skin and tadpole back skin collagenases, but they failed to inhibit the bacterial (Clostridium histolyticum) collagenase. The inhibitor was stable to heating to 60 degrees for 30 minutes and to trypsinization. PMID- 2992887 TI - A review of the role of cyclic GMP in neurological mutants with photoreceptor dysplasia. PMID- 2992888 TI - The pulmonary Kulchitsky cell (neuroendocrine) cancers: from carcinoid to small cell carcinomas. PMID- 2992889 TI - Mitral valve E-point septal separation as an indicator of ejection fraction in patients with reversed septal motion. AB - The mitral valve E-point septal separation (EPSS) is widely used as an M-mode echocardiographic indicator of normal or abnormal left ventricular ejection fraction. Other M-mode echocardiographic estimates of left ventricular function (eg, fractional shortening) have limited utility in the presence of abnormal septal motion. The utility of EPSS was investigated in predicting a normal or abnormal ejection fraction in 27 patients with reversed septal motion, and to compare these results with 103 patients with normal septal motion. It was determined that EPSS is valid as an indicator of the presence of a normal or abnormal ejection fraction regardless of abnormalities of septal motion. PMID- 2992890 TI - Pheochromocytoma and asthma. PMID- 2992891 TI - The dilemma of mild hypertension. Noninvasive evaluations of hypertensive patients. PMID- 2992892 TI - Sodium restriction amplifies and propranolol loading inhibits circadian rhythm of plasma renin-angiotensin and aldosterone. AB - Five male and 5 female clinically healthy volunteers, 17-37 years of age, gave systemic venous blood at 0600, 0800, 1200, 1800, 2000 and 0000 for RIA of (supine values) plasma renin-angiotensin (PRA) and aldosterone (PA) under 4 conditions: a. unrestricted sodium intake, no treatment; b. unrestricted sodium intake and 40 mg propranolol per os every 6 h; c. sodium restriction, no treatment; d. propranolol loading on sodium deprivation. Cosinor methods were used for data analysis. Sodium restriction amplifies the circadian rhythms of PRA and PA, whereas propranolol loading inhibits these same rhythms on a unrestricted sodium intake, but much less so under conditions of sodium deprivation. The propranolol induced inhibition of the circadian rhythms investigated on a unrestricted sodium intake suggests that the beta-adrenergic system is an effective mechanism coordinating the circadian rhythmic functions investigated. The persistence of the rhythms in sodium-depleted subjects under pharmacological blockade of beta adrenoreceptors is in keeping with the concept that a second mechanism of the circadian rhythms examined is located in the sodium-sensitive macula densa cells of the renal distal tubule. PMID- 2992893 TI - Effect of retinoids on rheumatoid arthritis, a proliferative and invasive non malignant disease. AB - In rheumatoid arthritis synovial tissue proliferates and destroys articular cartilage, bone and tendons. Collagenase is a major mediator of the connective tissue degradation. This enzyme is produced in large quantities by rheumatoid tissue and its synthesis can be inhibited by retinoids. However, knowledge of mechanisms controlling retinoid inhibition of collagenase production and of factors possibly controlling synovial cell proliferation is limited. We found that transforming growth factor beta in combination with epidermal growth factor, epidermal growth factor alone and immune interferon increased proliferation of cultured human and rabbit synovial fibroblasts. Only transforming growth factor beta caused a piling up of cells into foci resembling those seen in primary cultures of human rheumatoid tissue. All the factors were antagonized by retinoids but not by glucocorticoids or indomethacin. Adding retinoids or glucocorticoids to collagenase-producing cells decreased hybridizable collagenase mRNA by 50% within 24 h. Oral administration of retinoids to rats with experimental arthritis decreased clinical disease without toxicity, and inhibited collagenase synthesis by synovial cells taken from treated animals. Retinoids are both antiproliferative and anti-invasive, and therefore may be potential therapeutic agents in the treatment of rheumatoid arthritis. PMID- 2992894 TI - Role of retinoids in differentiation and growth of embryonal carcinoma cells. AB - To study how retinoids promote differentiation and inhibit proliferation of embryonal carcinoma (EC) cells, we have followed their intracellular fate. Retinoic acid (RA) is effectively metabolized to more polar compounds by many EC lines. Unlike RA, retinol is slowly metabolized. Our inability to detect conversion of retinol to RA might indicate that the two retinoids elicit their effects on EC cells in different ways. Retinol added to cultures quickly appears in the nuclear fraction; the proportion associated with nuclei after detergent extraction is initially very low but increases with time. Retinol and RA might be translocated to nuclei by their respective binding proteins [cellular retinol binding protein (CRBP) and cellular retinoic acid-binding protein (CRABP)]: isolated EC nuclei have specific, independent binding sites for both holoproteins but not their ligands. CRABP cannot be detected in the nucleoplasm of untreated EC cells, but activity is measurable after cells are exposed to RA. Interestingly, incubation with retinol promotes movement of both CRBP and CRABP into the nucleoplasmic fraction. Finally, we have demonstrated that brief exposure to RA dramatically reduces the cloning efficiency of EC cells. Since some cells are unaffected even by lengthy exposures to RA whereas the growth of their progeny is inhibited, we suggest that EC cells can become epigenetically refractory to RA. PMID- 2992895 TI - Disulfiram-induced hepatitis. Report of two cases and review of the literature. AB - Two cases of disulfiram-induced hepatitis are presented. Both cases had prompt recurrence of abnormal liver function tests after rechallenge with small doses of the drug. All cases of disulfiram-induced hepatotoxicity reported in the English literature are reviewed. PMID- 2992897 TI - [Transcription of the SV40 genome in Saccharomyces cerevisiae yeasts]. PMID- 2992896 TI - [Quaternary structure of NAD-kinase of rabbit liver during fermentation reactions]. PMID- 2992898 TI - 'Hangover' effects the morning after marijuana smoking. AB - Thirteen male marijuana smokers participated in a study to determine whether marijuana smoked in the evening would result in measurable subjective or other behavioral effects the following morning. Subjects smoked either active (2.9% delta 9THC) or placebo (0.0% delta 9THC) marijuana cigarettes according to a standardized smoking regimen. Smoke inhalation was monitored by measuring expired air carbon monoxide (CO) levels before and after smoking. Acutely, active marijuana produced significant changes in heart rate, CO level, various measures of subjective effects, and behavioral tasks of card sorting, free recall and time production. When the test battery was repeated the following morning (approx. 9 h after smoking), significant changes were observed on two subjective effects scales and on the time production task after active, but not placebo, marijuana. These apparent 'hangover' effects were different from the acute effects of marijuana. The findings suggest that marijuana smoking can produce residual (hangover) effects the day after smoking. The precise nature and extent of these effects, as well as their practical implications, remain to be determined. PMID- 2992899 TI - Evaluation of oral acyclovir therapy. AB - Acyclovir is a specific antiviral agent. The triphosphate form inhibits viral DNA replication by competing for incorporation into the replicating DNA chain or by inhibiting viral DNA polymerase. Cells not infected with herpesvirus are generally unaffected. Oral acyclovir inhibits most herpes simplex virus types 1 and 2, and varicella-zoster virus at concentrations used clinically. Oral acyclovir has an average plasma half-life of three hours and is eliminated primarily by renal mechanisms. Peak plasma concentrations occur 1.5 to 2.5 hours after administration and the oral bioavailability is 15 to 30 percent. Acyclovir distributes into most body tissues, including vesicular fluid and the central nervous system. Oral acyclovir is effective treatment of initial and recurrent genital herpes and can suppress frequently recurring genital herpes in both immunocompetent and immunocompromised patients. It is also effective for acute herpes zoster in the immunocompetent and possibly immunocompromised patient. No role is established in either Epstein-Barr virus or cytomegalovirus infections. Oral acyclovir appears to be effective and relatively safe, nontoxic therapy when administered in doses of 1-4 g/d. Oral acyclovir represents a major therapeutic advance in the treatment of herpesvirus infections. PMID- 2992900 TI - Hypersensitivity reaction to mannitol. AB - A 60-year-old atopic female was admitted for investigation of progressive neurological deficits involving her left hand, arm, and shoulder. A computed tomography scan demonstrated a right parietal lesion of her brain, which was removed. Increasing intracranial pressure necessitated the administration of mannitol 20% 250 ml. The patient stated she had a "tightness" in her chest during the mannitol administration. Five days later, rises in intracranial pressure again necessitated the infusion of mannitol 20% 150 ml. After 100 ml of the solution had been infused, the patient experienced mild respiratory distress, cyanosed lips, and hives of her abdomen. Supportive therapy, including aminophylline and diphenhydramine administration, abated her distress. Anaphylaxis to mannitol administration has only been reported twice previously, despite mannitol's widespread use. Physicians and pharmacists should be aware that severe hypersensitivity reactions may occur, especially in patients with a history of atopy. PMID- 2992901 TI - Effects of hypercholesterolemia on beta adrenoceptors in the rabbit heart. AB - Hypercholesterolemia has been shown to cause an increased vascular reactivity to catecholamines, but the mechanisms leading to greater smooth muscle responsiveness are not clear. Since membrane lipids are associated with receptors, a change in receptor structure and affinity for agonists could have been responsible. This study examined pharmacologic characteristics of cardiac beta adrenoceptors in isolated hearts from rabbits which had been fed a high cholesterol diet for 4, 6, or 8 weeks. Parameters measured were the spontaneous rate changes of the isolated right atrium and changes in refractory period of the isolated, perfused ventricles as these were induced by cumulative doses of the beta receptor agonist isoproterenol. The efficacy of a specific beta-one adrenoceptor blocking drug, metoprolol, in blocking these effects of isoproterenol was also evaluated. The rate and refractory period responses to isoproterenol and the blocking effectiveness of metoprolol were the same in all groups, as were the maximum responses induced by the agonist. It thus appears that hypercholesterolemia does not alter membrane beta adrenoceptor responsiveness in rabbit hearts. PMID- 2992902 TI - [Herpes simplex infection after kidney transplantation under immunosuppression by cyclosporin. Diagnostic and therapeutic experiences]. AB - Incidence and course of herpes simplex infections was determined prospectively in 22 patients who had a kidney transplant and were treated with cyclosporin. In addition to clinical findings, serial studies were undertaken of throat washings for herpes simplex virus in cell culture, as well as of patient sera for herpes specific IgG and IgM antibodies. There were 13 clinically manifest infections, 12 of them localized, while one had dissemination with necrotizing retinitis. Virus demonstration was successful in all cases in which virostatic drugs had not yet been used. Asymptomatic virus excretion was noted in three cases. Significant IgG titre rise occurred in six of the 13 cases, but a positive IgM titre in only two. Acyclovir proved to be an effective virostatic drug with few side effects. The outcome in the localized infections was favourable, but in the disseminated one residual defects remained. PMID- 2992903 TI - [Granular-cell tumor of the esophagus]. AB - A granular-cell oesophageal tumour was demonstrated in a 47-year-old man. With rare exception this type of tumour is benign. As it grows dysphagia develops. Surgical resection is the treatment of choice. It should be a wide excision, because the non-encapsulated tumour is microscopically more extensive than macroscopically. If too little is excised it may recur. But the high operative risk justifies expectant waiting, perhaps combined with local partial endoscopic removal of the tumour. Only in the rare instance of a small or pedicled tumour is total endoscopic excision possible. PMID- 2992904 TI - [Therapy of medullary thyroid carcinoma]. PMID- 2992905 TI - [Acute polyneuropathy caused by inhalation of solvent mixtures in varnish removers and thinners in normal commercial use]. AB - Three cases of acute intoxication after inhalation of solvents are reported. The clinical picture was that of a severe sensory-motor polyneuropathy, in one case with root involvement leading to complete Landry's paralysis with respiratory paralysis. All three patients required intensive care. The healing process was extraordinarily slow in two and was still unsatisfactory over a 1 1/2-year period of observation. The responsible agents were probably toluene and xylene. PMID- 2992907 TI - Excited-state controlled peroxide formation of DNA. AB - Reactions with water and 2 + 2-cycloadditions of individual bases are the primary steps held responsible for the deformation of DNA at short wavelengths. From the observed data however the influence of atmospheric oxygen on the light-induced reaction of DNA is evident. A plausible explanation for these effects is the formation of reactive oxygen species during the UV irradiation of DNA. In the present work the deformation of DNA by different oxygen species like singlet oxygen (1O2), superoxideanion (O2), hydroxyradical (OH.), and ozone (O3) is excluded with the help of chemical-trapping experiments. The photoinduced transformation proceeds via excited states of DNA, which react with oxygen to afford peroxide. PMID- 2992906 TI - Sialic acid residues on NGF receptors on PC12 cells. AB - Nerve growth factor (NGF) binding to cell surface receptors on PC12 cells is altered by the lectin wheat germ agglutinin (WGA), indicating that the receptor is a glycoprotein. Treatment of PC12 cells with most sugar-specific glycosidases does not substantially affect the ability of the receptor to bind NGF. High concentrations of N-acetyl-D-glucosaminidase (EC 3.2.1.30), however, decreased binding by 20-30%. The ratio of high- and low-affinity binding on PC12 cells also was not affected by glycosidase treatment. However, cleavage of sialic acid residues with neuraminidase (EC 2.3.1.18) increased the mobility of the NGF receptor complex in a sodium dodecyl sulfate polyacrylamide gel under reducing conditions. Covalent cross-linking of 125I-NGF to PC12 cells reveals the presence of two hormone-receptor complexes with molecular weights of 158,000 and 100,000 daltons, both of which underwent an 10,000 dalton apparent molecular weight decrease after neuraminidase exposure. Neuraminidase also affected the interaction of WGA with the receptor. WGA converted rapidly dissociating NGF binding into a slowly dissociating form, an effect which was inhibited by 50% by prior treatment with neuraminidase. Furthermore, a succinylated derivative of WGA, which binds N-acetyl-D-glucosamine but not sialic acid residues, unlike the native lectin, did not change the kinetic properties of the receptor. These results indicate that NGF receptors contain sialic acid residues which can interact with WGA producing in a change in receptor-binding properties. PMID- 2992908 TI - [Adrenergic mechanisms in the hypotensive action of a piperazine derivative of 2 aminotetralin]. PMID- 2992909 TI - [Catecholaminergic receptors of the blood cells of schizophrenics]. AB - The simultaneous determination of serum CA and their receptors in blood cells offers a possibility to evaluate disturbances of the DA and NA neuronal systems in man. High affinity binding sites for 3H-yohimbine in platelets, 3H-DHA on granulocytes and 3H-spiperone in lymphocytes from healthy control persons, unmedicated (n = 28), and medicated (n = 8) schizophrenics as well as from an unmedicated psychiatric control (n = 14) were investigated. Furthermore, the actual concentration of the circulating CA was determined with HPLC-ECD. In unmedicated schizophrenics as compared to controls, specific binding of 3H spiperone to lymphocytes was markedly elevated in capacity and less in affinity as compared to controls. For beta2-receptors a significant decrease was found in capacity with no change in affinity. The changes in alpha2-receptors with a slight decrease in capacity were less distinct. The concentrations of circulating CA ranged from normal values up to a more than 3 fold increase in NA and DA, whereas A concentrations were practically unchanged. No overall change in these data was found in the medicated schizophrenic patients. 3H-spiperone binding was characteristically increased only in schizophrenics, but did not elevate above control data in the non-schizophrenic psychiatric control group. Preliminary experiments of family studies suggest that this model could be valuable as a vulnerability marker. PMID- 2992910 TI - Effects of adrenalectomy and dexamethasone administration on the level of prepro corticotropin-releasing factor messenger ribonucleic acid (mRNA) in the hypothalamus and adrenocorticotropin/beta-lipotropin precursor mRNA in the pituitary in rats. AB - RNA blot hybridization analysis with cloned rat CRF precursor (prepro-CRF) cDNA as a probe showed that prepro-CRF mRNA existed in rat hypothalamic and extrahypothalamic brain tissue, whereas it was undetectable in the pituitary and adrenal. To study the effect of glucocorticoid on the level of prepro-CRF mRNA in the hypothalmus and that of ACTH/beta-lipotropin (beta LPH) precursor mRNA in the pituitary, effects of adrenalectomy and dexamethasone administration were studied in rats. Adrenalectomy markedly raised mRNA coding for ACTH/beta LPH precursor in the anterior pituitary, but not in the neurointermediate pituitary lobe. Hypothalamic pre-pro-CRF mRNA increased only to 152% of the control value, 7 days after adrenalectomy. The administration of dexamethasone (200 micrograms/day for 7 days) started immediately after adrenalectomy lowered the ACTH/beta LPH precursor mRNA level in the anterior pituitary to 19% of the intact control value, whereas the level of prepro-CRF mRNA in the hypothalamus decreased only to 102%. These results suggest that glucocorticoids exert their feedback effect at the level of gene expression on both hypothalamic CRF neurons and pituitary corticotropes. Although the possibility that CRF neurons insensitive to glucocorticoid in the hypothalamus might blunt the change in the prepro-CRF mRNA could not be ruled out, it is also possible that the effect of glucocorticoids on the pituitary is dominant. PMID- 2992911 TI - Angiotensin-converting enzyme in the testis and epididymis: differential development and pituitary regulation of isozymes. AB - Angiotensin-converting enzyme (ACE, EC 3.14.5.1) is found in particulate fractions of the epididymis but not in soluble epididymal fractions or in the testis of 4-week-old rats. [3H]Captopril autoradiography of testis and epididymis from 4-week-old rats confirms the association of ACE with epididymal ducts but not the testis. ACE appears in the testis between 4 and 6 weeks of age. Soluble ACE is not detectable in the epididymis until 6-7 weeks of age. Within the epididymis, regions closest to the testis develop soluble ACE activity about 1 week before those nearest to the vas deferens. Hypophysectomy of 10 week-old-rats depletes greater than 95% of ACE activity from the testis and soluble fractions of the epididymis, with little change in ACE levels from particulate fractions of the epididymis. [3H]Captopril autoradiography after hypophysectomy reveals luminal and epithelial ACE in the epididymis. The presence of particulate ACE in the epididymis under conditions where there is no testicular ACE indicates that the two forms are synthesized separately. However, soluble ACE from the epididymis might be derived from the membrane-associated ACE of the testis. Such a relationship is supported by the lag of 1 week between the development of ACE in the initial segment of the epididymis and the tail of the epididymis, and by the occurrence of soluble epididymis ACE only in those animals with testicular ACE activity. PMID- 2992912 TI - Evidence that beta-endorphin binds to specific receptors in rat peripheral tissues and stimulates the adenylate cyclase-adenosine 3',5'-monophosphate system. AB - With the use of [125I]acetyl human beta-endorphin (Ac-hBE), specific binding sites for beta-endorphin (BE) were identified in the liver, kidney, adrenal, spleen, and testis of adult male rats, whereas specific BE-binding sites were not present in the ventral prostate or pancreas. In those tissues containing specific BE-binding sites, microsomal membranes (15,000-100,000 X g pellet) exhibited higher BE-binding capacity than the crude homogenate (125-100,000 X g pellet). The binding of BE was saturable, and maximal, specific binding was achieved with a 60-min incubation at 22 C. Furthermore, optimal BE binding was dependent on the presence of magnesium chloride. Scatchard analysis of BE binding to hepatic membranes revealed the existence of two classes of binding sites. One class had an apparent Ka of 0.019 X 10(9) M-1 and a lower number of binding sites (9.1 pmol BE/mg protein), whereas the other class had a lower affinity (apparent Ka of 0.0006 X 10(9) M-1) and a higher number of binding sites (159 pmol/mg protein). Specific BE binding to hepatic membranes was inhibited (80-100%) by rat AcBE-(1 27) and -(1-31), nonacetylated rat BE-(1-31), and human beta-lipotropin. At substantially higher peptide concentrations (greater than 10(-5) M), gamma endorphin, met-enkephalin, or leu-enkephalin inhibited BE binding by 20-40%. In addition, opiate receptor-binding drugs, such as morphine and naloxone, at 10(-5) M did not alter BE binding to hepatic membranes. Incubation of hepatic membranes with BE induced a dose-related increase in membrane adenylate cyclase activity, and 0.5 X 10(-10) M BE resulted in a maximal enhancement of adenylate cyclase activity to 148% above control values. Water-deprived or salt-loaded male rats with chronically lowered immunoreactive plasma BE exhibited substantially increased BE binding to adrenal and kidney tissue. Specific binding sites for BE occur in a variety of peripheral tissues, and alterations of circulating BE result in changes in the capacity of certain peripheral tissues to bind BE. Finally, occupancy of specific BE-binding sites in peripheral tissue stimulates the adenylate cyclase-cAMP system, which suggests that the peripheral actions of circulating BE may be mediated via this system. PMID- 2992914 TI - 1,2-Didecanoylglycerol and phorbol 12,13-dibutyrate enhance anterior pituitary hormone secretion in vitro. AB - Experiments were designed to evaluate the role of activators of protein kinase C, such as 1,2-diacylglycerol and phorbol esters, on the release of all the anterior pituitary (AP) hormones in vitro. Dispersed rat AP cells were incubated in the presence of 1,2-didecanoylglycerol (DiC10), a synthetic diacylglycerol, or phorbol 12,13-dibutyrate (PDBu), a tumor-promoting phorbol ester, at different concentrations and for varying periods of time. ACTH and beta-endorphin (beta End) secretion were enhanced by DiC10 in a concentration-dependent manner, with a minimal effective concentration of 5 microM. PDBu at 5 nM produced a significant release of both ACTH and beta-End. The effect of DiC10 and PDBu was time dependent, with maximal responses occurring at 15-30 min for DiC10 and 30-60 min for PDBu. Release of GH was also enhanced significantly by DiC10 and PDBu, with minimal effective concentrations of 1 microM and 1 nM, respectively. Maximal release of GH was already attained within 15 min with DiC10 or 60 min with PDBu. In additional experiments, the effects of DiC10 and PDBu on secretion of LH, FSH, PRL, and TSH were evaluated. The results indicate that 5-25 microM DiC10 produced a concentration-dependent release of each of those hormones, and that 5 microM was the minimal effective concentration in every case. Nearly maximal stimulation was achieved within 15 min for each hormone. PDBu (50 nM) significantly enhanced LH, FSH, PRL, and TSH release within 30 min. Although qualitatively all hormones were similarly stimulated, both with respect to time and concentration, some quantitative differences were observed. ACTH and beta-End release were enhanced 100% by DiC10 and 300% by PDBu, whereas the increase in other hormones was of a lesser magnitude. The present study indicates that two specific stimulators of protein kinase C, diacylglycerol and phorbol ester, can enhance secretion of all AP hormones in a concentration- and time-dependent manner. This suggests that formation of endogenous 1,2-diacylglycerol may represent a physiological intracellular messenger in the events leading to AP peptide hormone release. PMID- 2992913 TI - Microsomal phosphorylase in rat heart: depletion following adrenalectomy and restoration by in vivo administration of dexamethasone. AB - Phosphorylase activities were determined in homogenates, particulate and soluble fractions of heart muscle from control, adrenalectomized, and adrenalectomized dexamethasone-treated rats so as to assess the influence, if any, of glucocorticoids on the subcellular distribution of this enzyme. The specific activities (enzyme activity expressed as units per mg protein) and recoveries (enzyme activity expressed as units per fraction/g tissue) of both the active form of phosphorylase (phosphorylase a) and total phosphorylase (phosphorylases a and b) were essentially similar in homogenates and 10,000 X g particulate fractions prepared from hearts of control, adrenalectomized, and adrenalectomized dexamethasone-treated rats. Interestingly, however, the specific activities and recoveries of active as well as total phosphorylase were markedly lower (70-80%; P less than 0.001) in cardiac microsomes of adrenalectomized compared to control rats. On the other hand, the specific activities and recoveries of phosphorylase (active and total) were significantly higher (30-60%; P less than 0.01 or 0.001) in the cytosol fraction of heart muscle from adrenalectomized compared to control rats. Treatment of adrenalectomized rats with dexamethasone (a synthetic glucocorticoid) restored the specific activities and recoveries of phosphorylase in heart microsomal and cytosol fractions to control levels. No appreciable differences were observed in the specific activity ratios of phosphorylase a to phosphorylases a and b in any of the subcellular fractions of cardiac muscle from control, adrenalectomized, and adrenalectomized dexamethasone-treated rats. The above findings suggest a potential involvement of glucocorticoids in the maintenance of the membrane-associated (microsomal) pool of phosphorylase and, therefore, in the cellular compartmentalization of this enzyme in heart muscle. PMID- 2992915 TI - Adenylate cyclase in the corpus luteum of the rhesus monkey. III. Changes in basal and gonadotropin-sensitive activities during the luteal phase of the menstrual cycle. AB - The activity of adenylate cyclase was examined in corpora lutea (CL) obtained from rhesus monkeys at specific stages in the luteal phase of the menstrual cycle [3-5, 6-8, 9-12, 13-15, and 16 days (menses) after the midcycle LH surge]. The conversion of [alpha-32P]ATP to [32P]cAMP was used to monitor adenylate cyclase activity. cAMP production in luteal homogenates was assessed in the absence (basal activity) and presence of maximum stimulatory doses of forskolin (100 microM), 5'-guanylylimidodiphosphate [GMP-P(NH)P; 50 microM], GTP (50 microM), and GTP plus increasing doses of hLH and hCG. Basal activity was low in the early luteal phase (days 3-5; mean +/- SE, 1.2 +/- 0.2 pmol cAMP/mg protein X min), increased (P less than 0.05) by the midluteal phase (days 6-8 and 9-12, 2.1 +/- 0.4 and 2.0 +/- 0.3 pmol/mg X min, respectively), and then declined (P less than 0.05) during the late luteal phase (days 13-15 and 16-menses, 1.6 +/- 0.3 and 1.2 +/- 0.5 pmol/mg X min, respectively). Activity stimulated by GTP and GMP-P(NH)P [e.g. GMP-P(NH)P approximately 12 times basal level] followed the same pattern as basal activity during the luteal phase. In contrast, cAMP production in the presence of forskolin did not change significantly throughout the luteal phase. In the midluteal phase (days 6-8 and 9-12; n = 12), hCG and human LH (hLH) stimulated adenylate cyclase in a similar dose-dependent manner. Maximal stimulation of cAMP production by hCG was about 10% greater (P less than 0.05) than that by hLH; the activation constant was 12.3 nM for hCG and 28.3 nM for hLH. The maximal response to hLH and hCG as well as the sensitivity of adenylate cyclase to activation by hLH were greater (P less than 0.05) in the midluteal phase than in the early or late luteal phase. Decreased basal, gonadotropin stimulated, and guanine nucleotide-stimulated cAMP production and diminished sensitivity of adenylate cyclase to hLH correlated with a decline (P less than 0.05) in circulating progesterone and luteal weight during the late luteal phase. Thus, the adenylate cyclase system of the rhesus monkey CL undergoes significant changes during the luteal phase which are associated with the development and regression of the CL of the menstrual cycle. Mechanisms that modulate gonadotropin and nucleotide activation of adenylate cyclase without interfering directly with the catalytic unit are implicated in the changes that accompany luteolysis. PMID- 2992916 TI - Binding of intact parathyroid hormone to rat osteosarcoma cells: major contribution of binding sites for the carboxyl-terminal region of the hormone. AB - Most studies of PTH receptor binding have been carried out with amino-terminal radioligands which only detect binding within that region of the hormone molecule. We studied the binding of electrolytically labeled intact bovine PTH [bPTH-(1-84)], and its amino-terminal fragment, bPTH-(1-34) to intact cloned rat osteosarcoma cells (ROS 17/2.8). We also measured the effects of these hormones on cell cAMP accumulation. Binding equilibrium for the two radioligands was reached by 2 h of incubation at 22 C. However, the cells had higher binding capacity (8-9% or 0.22-0.25 fmol/2 X 10(6) cells) for [125I]bPTH-(1-84) than for [125I]bPTH-(1-34) (4% or 0.11 fmol/2 X 10(6) cells). On the other hand [125I]bPTH (1-34) bound to ROS cells with higher affinity [dissociation constant (Kd) = 19 nM] than did [125I]bPTH-(1-84) (Kd = 210 nM). Measurements of trichloroacetic acid precipitability and analysis of rebinding of previously incubated radioligand to fresh cells ruled out degradation of the tracer as an explanation for these differences. The maximum cell cAMP response to bPTH-(1-34) (Vmax = 780 +/- 32 pmol/2 X 10(6) cells X 5 min) was reached at 10(-7) M concentration with an affinity [Michaelis-Menten constant (Km)] of 3 nM. On the other hand, the Vmax with intact bPTH-(1-84) was lower (400 +/- 7 pmol/2 X 10(6) cells X 5 min), with a Km of 60 nM). Further studies with the bPTH-(1-84) tracer showed inability of hormonal fragments to compete completely for binding. At a concentration of 3 microM, bPTH-(1-84) reduced tracer binding by 82.5%, compared to 18% by bPTH-(1 34) and 10% by (Nle8,Nle18,Tyr34)bPTH-(1-34)amide, 60% by human PTH (hPTH)-(53 84), and 70% by the combination of bPTH-(1-34) and hPTH-(53-84). hPTH-(53-84) itself did not elicit a cAMP response after 5 min or 1 h of incubation nor did it significantly alter the cAMP response of the cells to bPTH-(1-84). These studies suggest that PTH binds to ROS 17/2.8 cells by sites carboxy-terminal (C-terminal) to position 34, in addition to sites within the amino-terminal portion of the hormone molecule; 72% of the binding of intact hormone to these cells was to the C-terminal 35-84 region of the PTH molecule. The significance of the C-terminal binding sites is presently unclear, but they do not appear to be coupled to adenylate cyclase. Further work is needed to determine the effects of C-terminal PTH fragments on bone cell metabolism. PMID- 2992917 TI - Multisite inhibition by ipodate of iodothyronine secretion from perfused dog thyroid lobes. AB - Cholecystographic radiocontrast agents interfere with thyroid hormones in several ways. In the present study 1 mM ipodate induced a rapid sustained and reversible inhibition of the secretion of T4, T3, rT3, 3,3'-diiodothyronine, and 3',5' diiodothyronine from perfused dog thyroid lobes. This effect was not reproduced by infusion of 3 mM iodide and not affected by 2 mM methimazol or 2 mM perchlorate. One millimolar of ipodate inhibited secretion of T4 to 23.7 +/- 2.8% of control (+/- SE, n = 6), 0.3 mM ipodate to 59.6 +/- 3.01 (n = 4), and 0.1 mM ipodate to 80.4 +/- 5.7% of control (n = 4). In search of the site of action in the thyroid of this inhibitory compound it was found that 1 mM ipodate inhibited TSH-induced increase in thyroidal cAMP, cAMP-induced generation of intracellular colloid droplets, and liberation of T4 and T3 from thyroglobulin by acid proteases and peptidases. These processes are those thought to be inhibited during iodide inhibition of thyroid secretion, via gradual formation of an unknown iodine-containing organic intermediate. It is suggested that the inhibition of thyroid secretion observed in the present study is due to structural similarities between ipodate and this putative iodine-containing mediator of the iodide-induced inhibition of thyroid secretion. PMID- 2992918 TI - External root resorption following bleaching of pulpless teeth with oxygen peroxide. PMID- 2992919 TI - Insulin-like growth factors as intraovarian regulators of granulosa cell growth and function. AB - A relatively large body of evidence now appears to support the existence of the essential ingredients for novel intraovarian IGF-driven control mechanisms. Indeed, evidence presented in this communication is in keeping with the possibility that the granulosa cell may be the site of IGF production, reception, and action. Although the relevance of IGFs to ovarian cell types other than the granulosa cell is largely unknown, one cannot at the present time exclude the possibility of nongranulosa cell contributions to intraovarian IGF production, reception, and action. Indeed, preliminary affinity cross-linking studies (Adashi, Resnick, Svoboda, Van Wyk and D'Ercole; unpublished data) suggest the existence of type-I and type-II receptors in nongranulosa cell compartments. The above notwithstanding, IGFs of granulosa (and possibly circulatory) origins may interact with granulosa cell autoreceptors either independently or in synergy with other granulosa cell agonists. According to this view, IGFs may act in the autocrine mode to stimulate granulosa cell replication on the one hand and promote granulosa cell differentiation on the other. Although proliferation and terminal differentiation may prove mutually exclusive under some circumstances, coexistence of the two processes is being increasingly recognized. In this context, some studies of porcine granulosa cells support a dual role for IGFs in granulosa cell ontogeny. As such, the IGFs can be added to a growing list of growth factors known to modulate granulosa cell growth and function, including EGF, PDGF, and FGF. Our findings indicate that Sm-C/IGF-I synergizes with FSH in the induction of rat granulosa cell aromatase activity at nanomolar concentrations compatible with its granulosa cell receptor binding affinity (thus far studied only in porcine cells. A role for Sm-C/IGF-I in the regulation of this key granulosa cell function would be in keeping with the possibility that Sm C/IGF-I may partake in the assertion and maintenance of dominance by the selected follicle(s) or in promoting juvenile and early follicular development. Moreover, the ability of Sm-C/IGF-I to potentiate this and other FSH-driven ovarian functions may also account, at least in part, for the puberty-promoting effect of growth hormone. This permissive action of growth hormone has been initially suggested by observation in growth hormone-deficient rats, mice (dwarf mutants, and humans (sporadic, hereditary or acquired growth hormone deficiency.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2992920 TI - Polybrominated biphenyls in model and environmentally contaminated human blood: protein binding and immunotoxicological studies. AB - A review and summary is given of analytical, biochemical, and immunological studies made following an immunodiagnostic investigation which revealed significant decreases in the numbers, and changes in the functional integrity, of both T-and B-lymphocytes in a group of Michigan dairy farmers exposed to polybrominated biphenyls (PBB) which had been inadvertently introduced into the food chain in 1973. A quantification technique based on selected ion monitoring of bromine anions, obtained in negative chemical ionization, permitted determination of 10-35 pg of individual PBB congener per mL serum, a 20-fold improvement over electron capture gas chromatography. An in vitro spiked system was established and shown to be a representative model of environmentally contaminated blood. Immunoprecipitation followed by mass spectrometric quantification determined that the distribution of PBB among plasma, erythrocytes, mononucleocytes and polymorphonucleocytes was 89:9:less than 1:less than 1. In plasma 80% of the PBB was bound to apolipoproteins B and A in a 3:1 ratio. No preferential absorption of PBB congeners was found in the blood compartments suggesting that changes in the relative abundances of PBB congeners observed in longitudinal studies on Michigan subjects reflect differences in excretion rates or metabolism. A repeat in 1981 of the immunodiagnostic tests conducted in 1976 revealed a virtually complete persistence of the immune dysfunctions in the Michigan farmers exposed to PBB a decade ago. PMID- 2992921 TI - Utilization of adipose tissue biopsy in characterizing human halogenated hydrocarbon exposure. AB - Halogenated hydrocarbons have been successfully utilized for pest control in agriculture and public health. In industry, the polychlorinated biphenyls (PCBs) have been particularly useful. Unfortunately, residues have proven persistent and have been found widely dispersed in the environment. Because they have chemical properties that favor bioaccumulation, it is not surprising that many have been identified in animals and man. Such findings prompted public health concern and initiated review of potential adverse health impacts. In many instances this process has led to total prohibition of use or strict limitations. Highly lipophilic, the primary accumulation site in humans is adipose tissue. Analysis of adipose samples remains the preferred biologic index for estimating exposure. Improved instrumentation has lowered the limit of detection and improved the accuracy of quantification. Accumulated population experience has helped develop understanding of the dynamics of tissue partitioning in humans. Once such relationships have been adequately described, other less invasive procedures may be more confidently used for general surveillance purposes. Whenever adipose tissue is obtained, the blood compartment should also be sampled and the relationship reported. We have only begun to investigate the resulting partition ratio as an investigative tool. PMID- 2992922 TI - Symptoms, signs and findings in humans exposed to PCBs and their derivatives. AB - The records of the health effects caused by some accidental exposure and findings from medical examination in cases exposed to polybrominated biphenyls (PBB) as well as to polychlorinated biphenyls (PCB) and their derivatives polychlorinated dibenzofurans (PCDF) and dibenzodioxins (PCDD) have provided some information for the recognition and classification of their toxicity in humans. The most impressive clinical features have been presented by the yusho episode of exposure. Dermatologic signs are the most persistent indicator of a considerable uptake. Neurological symptoms, respiratory findings and impairment of liver function are further aspects of the contamination. Skin manifestations have been observed also in the newborn infants from mothers exposed to high levels of the substances. However, the available data make it still hard to assess the clinical picture of the effects on humans in cases of acute exposure and even more the effects on reproduction and long-term effects. Furthermore it would still be arbitrary to draw a line between the symptoms which can be referred to PCBs and PBBs alone and those which can be related qualitatively and quantitatively to PCB derivatives (PCDFs, PCDDs, PCQs). PMID- 2992923 TI - Comparative toxicity of PCBs and related compounds in various species of animals. AB - There are several basic principles that apply to the clinicopathologic syndrome produced by polychlorinated biphenyls (PCBs). They are as follows: The degree of halogenation and position of the halogen atoms determine the potency of PCB, PBB, CDD, CDF and CN; in a given species of animals, the clinicopathologic syndrome induced by PCB is comparable to that induced by polybrominated biphenyls (PBB), chlorinated dibenzo-p-dioxins (CDD), chlorinated dibenzofurans (CDF), and chlorinated naphthalenes (CN) when an equitoxic dose is achieved; The clinicopathologic syndrome is different in each species of animals; Different species of animals vary in their susceptibility to intoxication; intoxication is more readily effected in young animals that in adults; at lethal doses the time between exposure and death is prolonged (greater than 2 weeks). PMID- 2992924 TI - Effects of PCBs and related compounds on hepatocarcinogenesis in rats and mice. AB - Commercial mixtures of polychlorinated biphenyls (PCBs) and polybrominated biphenyls (PBBs) can cause hepatocellular carcinoma in rats and mice. Present evidence indicates that these chemicals act as promoters and not initiators of hepatocarcinogenesis. Our results show that Firemaster BP-6 (FM) and its nontoxic major congener, 2,2', 4,4', 5,5'-hexabromobiphenyl (HBB), act as promoters in the two-stage model for hepatocarcinogenesis devised by Pitot and associates. A toxic congener, 3,3', 4,4', 5,5'-HBB, also was assessed for tumor-promoting activity. This congener, though not in FM, is similar to TCDD, in that both cause 3 methylcholanthrene (MC)-type induction of hepatic microsomal enzymes and produce similar toxic responses. FM contains several congeners which are mixed-type inducers in that they induce MC-type and phenobarbital (PB)-type enzymes. The toxicity of FM is most likely associated with its congeners which are mixed-type inducers and not to relatively nontoxic congeners such as 2,2', 4,4', 5,5'-HBB which are strictly PB-type inducers. Congener 3,3', 4,4', 5,5'-HBB acted as a tumor promoter only at a dose that was hepatotoxic. A synergistic effect on tumor promoting ability was produced by combining a nontoxic and nonpromoting dose of 3,3', 4,4', 5,5'-HBB with a promoting dose of 2,2', 4,4', 5,5'-HBB. Our results suggest that synergism between toxic and nontoxic congeners in FM may explain why mixtures such as FM have greater promoting ability than individual congeners. Our results also indicate that with PBB, toxicity and carcinogenicity are not necessarily related. PMID- 2992925 TI - Persistence of polybrominated biphenyls (PBB) in human post-mortem tissue. AB - Polybrominated biphenyl (PBB), a fire retardant, was accidentally substituted for an animal feed supplement in Michigan 10 years ago. This led to widespread livestock contamination and ultimately to contamination of virtually every human residing in the State at that time. In order to evaluate the extent of PBB persistence and distribution in human tissues 10 years after this accidental poisoning, a study was conducted on a series of autopsy cases from the Grand Rapids ("high" exposure) area of the State. No attempt was made to relate cause of death to PBB exposure or tissue concentration. Samples of 196 tissues from 15 subjects were analyzed for PBB content, and levels were determined by measurement of the hexabromobiphenyl peak using electron capture gas chromatography. Only 4 of the 196 samples analyzed did not have PBB concentration above the limit of detection (0.5 ng/g). As expected, fat and fat-rich tissue had the highest PBB concentration. Perirenal fat had the highest mean concentration (475 ng/g). Adrenal, atheromatus aorta and thymus had mean concentrations about half that of perirenal fat; all other tissues had mean concentrations one-tenth or less of perirenal fat. The results document that PBB is still present in human tissue and that PBB was distributed in all tissues examined. The PBB fat elimination half time was estimated to be at least 7.8 years. If this is approximately correct, PBB will persist in tissues throughout the lifetime of humans so contaminated. PMID- 2992926 TI - Mechanisms of the biological effects of PCBs, polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in experimental animals. AB - Polychlorinated biphenyls, certain polychlorinated dibenzo-p-dioxins and certain polychlorinated dibenzofurans cause a variety of biological effects in experimental animals. The mechanism of the induction of certain enzymes is perhaps best understood. That is, there is binding of certain chlorinated biphenyls, dibenzo-p-dioxins and dibenzofurans to a receptor, translocation of the compound-receptor complex into the nucleus followed by an increased activity of a number of enzymes in the cell. Although the concentration of this receptor in various tissues of some mouse strains correlates well with the intensity of some of the biological effects observed in the mouse strains exposed to these compounds, this correlation apparently does not extend across various species. The current evidence suggests that the acute toxic effects of TCDD in various species is in some way associated with binding of TCDD to the receptor. However, biological effects of TCDD in addition to those resulting from binding to the receptor may be required to produce acute toxicity and, perhaps, other effects. The acute toxic effects of TCDD are probably caused by the parent compound rather than metabolites; however, this conclusion must be viewed as tentative. Also, it cannot be excluded at this time that biological effects other than acute toxicity may be caused by metabolites of TCDD. Finally, the acute toxic effects of TCDD appear not to be related, at least not directly, to the rate of metabolism of TCDD in experimental animals nor to the half-life of excretion. PMID- 2992928 TI - Structure-induction versus structure-toxicity relationships for polychlorinated biphenyls and related aromatic hydrocarbons. AB - A comparison of the structure-induction (involving rat and mouse Ah receptor binding) and structure-toxicity (in vivo guinea pig toxicity) relationships suggests that two receptors with structurally distinct binding properties may be involved. This is supported by demonstration of potentiated toxicity through a mechanism believed to involve the Ah receptor as a site of loss with respect to toxicity. Theoretical and working models are proposed for these separate receptors to aid in the search for other relevant binding proteins. The findings suggest that polychlorinated biphenyls that are relatively low in toxicity may have modulating properties on the action of highly toxic compounds with which they are normally found in the environment. PMID- 2992927 TI - PCBs: structure-function relationships and mechanism of action. AB - Numerous reports have illustrated the versatility of polychlorinated biphenyls (PCBs) and related halogenated aromatics as inducers of drug-metabolizing enzymes and the activity of individual compounds are remarkably dependent on structure. The most active PCB congeners, 3,4,4',5-tetra-, 3,3',4,4'-tetra-, 3,3',4,4',5 penta- and 3,3',4,4',5,5'-hexachlorobiphenyl, are substituted at both para and at two or more meta positions. The four coplanar PCBs resembled 3-methylcholanthrene (3-MC) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) in their mode of induction of the hepatic drug-metabolizing enzymes. These compounds induced rat hepatic microsomal benzo(a)pyrene hydroxylase (aryl hydrocarbon hydroxylase, AHH) and cytochromes P-450a, P-450c and P-450d. 3,4,4',5-Tetrachlorobiphenyl, the least active coplanar PCB, also induced dimethylaminoantipyrine N-demethylase and cytochromes P-450b+e and resembled Aroclor 1254 as an inducer of the mixed function oxidase system. Like Aroclor 1254, all the mono-ortho- and at least eight di-ortho-chloro analogs of the coplanar PCBs exhibited a "mixed-type" induction pattern and induced microsomal AHH, dimethylaminoantipyrine NM demethylase and cytochromes P-450a-P-450e. Quantitative structure-activity relationships (QSARs) within this series of PCBs were determined by comparing their AHH induction potencies (EC50) in rat hepatoma H-4-II-E cells and their binding affinities (ED50) for the 2,3,7,8-TCDD cytosolic receptor protein. The results showed that there was an excellent correlation between AHH induction potencies and receptor binding avidities of these compounds and the order of activity was coplanar PCBs (3,3',4,4' -tetra-, 3,3',4,4',5-penta- and 3,3',4,4',5,5'-hexachlorobiphenyls) greater than 3,4,4',5-tetrachlorobiphenyl approximately mono-ortho coplanar PCBs greater than di-ortho coplanar PCBs. It was also apparent that the relative toxicities of this group of PCBs paralleled their biological potencies. The coplanar and mono-ortho coplanar PCBs also exhibit differential effects in the inbred C57BL/6J and DBA/2J mice. These compounds induce AHH and cause thymic atrophy in the former "responsive" mice whereas at comparable or higher doses none of these effects are observed in the nonresponsive DBD/2J mice. Since the responsiveness of these two mice strains is due to the presence of the Ah receptor protein in the C57BL/6J mice and its relatively low concentration in the DBA/2J mice, the results for the PCB cogeners support the proposed receptor-mediated mechanism of action. PMID- 2992929 TI - Toxicity of chlorinated aromatic compounds in animals and humans: in vitro approaches to toxic mechanisms and risk assessment. AB - Human exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and chlorinated analogs commonly results in pathological changes in the skin and its appendages characterized by thickening of the epidermis (acanthosis), hyperkeratosis and squamous metaplasia of the epithelial lining of the sebaceous glands. Acneform lesions (chloracne) develop as hair follicles dilate and fill with keratin and sebaceous glands become cystic. In animal models it has been found that the chloracneogenic potential of the halogenated aromatic compounds examined corresponds with the relative affinity of these same compounds for the cytosolic TCDD receptor. This receptor controls the coordinate expression of a number of inducible enzyme activities and in certain cell targets can alter normal programs of proliferation and differentiation. In this report we describe some of our ongoing studies on the mechanisms of action of TCDD in normal human epidermal cells and squamous cell carcinoma (SCC) lines. These systems permit detailed investigation of the molecular and biochemical events underlying pathologic changes in the skin and offer the potential of establishing a risk assessment model for halogenated aromatic compounds by using human target cells. PMID- 2992930 TI - Pharmacological characterization of two specific binding sites for neurohypophyseal hormones in hippocampal synaptic plasma membranes of the rat. AB - Synaptic plasma membranes containing binding sites for tritiated oxytocin and arginine vasopressin were isolated from rat hippocampus. The binding parameters for oxytocin and vasopressin sites were determined and statistically analysed. The fitted curve for oxytocin binding was compatible with a model where the ligand interacts with two classes of receptors with different capacities and affinities. The sites with low binding capacity had an apparent dissociation constant at equilibrium of 1.8 nM and a maximal binding capacity of 17 fmol/mg protein. By contrast, the Scatchard plot failed to reveal a marked heterogeneity in the population of sites labelled with [3H]vasopressin with an affinity of 1.5 nM and a maximal binding capacity of 39 fmol/mg protein. The specificity of these binding sites, tested in competition experiments, revealed that these neurohypophyseal hormones labelled two distinct populations of sites. One population with a high affinity for vasopressin, oxytocin and vasotocin, the other population with a high affinity for vasopressin and vasotocin and a low affinity for oxytocin. Adenylate cyclase activity was not affected by arginine vasopressin or oxytocin. These receptors are compared with previously characterized peripheral receptors. PMID- 2992931 TI - Chromosome translocations clustered 5' of the murine c-myc gene qualitatively affect promoter usage: implications for the site of normal c-myc regulation. AB - Five novel murine plasma cell (PC) tumors with chromosome translocations 350-500 bp 5' of the first c-myc exon are described. The t(12;15)s of TEPC 1194, ABPC 33 and TEPC 1165 position the intact c-myc locus 5' of the Cu, C gamma 2a and C alpha IgCH genes respectively. In ABPC 17, the IgH enhancer element and adjacent switch (Su) sequences were found 5' of the first c-myc exon while this enhancer is associated with the reciprocal products of the TEPC 1194, ABPC 33 and TEPC 1033 translocations. Quantitative S1 nuclease analyses demonstrate that the ratios of transcription from the two c-myc promoters (P1 and P2) are increased 4 to 7-fold in these five tumors. With the exception of TEPC 1165, (which contains a small deletion in exon 1), such increases in P1:P2 ratios appear to be manifested by a reduction in P2 usage in comparison to other tumors without such promoter shifts. A survey of 27 additional PC and non-PC B lymphoid tumors and cell lines revealed that myc promoter shifts of this magnitude are unique to PC tumors with 5'-proximal translocations. We propose that (i) these clustered breakpoints identify a normal c-myc regulatory element located at least 350 bp 5' of c-myc exon 1; (ii) the loss or disruption of this cis-acting upstream element and the linkage of c-myc to the IgCH locus would result in abnormal expression of this oncogene in these as well as most other PC tumors. PMID- 2992932 TI - Isolation and characterization of a human T cell leukemia virus type II from a hemophilia-A patient with pancytopenia. AB - Human T cell leukemia virus (HTLV) type I has been isolated from the cultured T cells of several patients with adult T cell leukemia (ATL) and has been etiologically linked to ATL. However, HTLV-type II has been isolated only once, from the T cells of a patient with a T cell variant of hairy-cell leukemia. We report here the isolation of HTLV-II-related virus from the cultured T cells of a hemophilia-A patient with pancytopenia. The T cell line (CM) grows in the absence of T cell growth factor. Cord blood T cells were rapidly transformed when co cultivated with irradiated CM cells. Heterologous competition radioimmunoassays using purified HTLV-I p24 showed the expression of HTLV-IIMO-related protein in these cells. Electron microscopy of the CM cells showed the presence of intracellular and extracellular type C viral particles. Comparison of the proviral genome in the CM cell line and the prototype HTLV-IIMO-containing cell line (MO) by molecular hybridization with probes specific for HTLV-IIMO indicated that restriction cleavage sites were identical. The fresh peripheral blood leukocytes of the patient contained two complete copies of the proviral genome, despite the lack of HTLV-II p24 expression. The virus from the cell line CM is designated as HTLV-IICM to distinguish it from the original HTLV-IIMO isolate. PMID- 2992933 TI - Glutamic acid decarboxylase activity is stimulated in quail retina neuronal cells transformed by Rous sarcoma virus and is regulated by pp60v-src. AB - Rous sarcoma virus (RSV) stimulates in quail embryo neuro-retina (NR) cultures the specific activity of glutamic acid decarboxylase (GAD), the enzyme responsible for the synthesis of gamma-aminobutyric acid, a major inhibitory neurotransmitter in NR and in central nervous system. In quail embryo NR cultures transformed by ts NY-68, a thermodependent transformation-defective mutant of RSV, stimulation of GAD activity is regulated by pp60v-src, the product of the src gene of RSV. Fibroblasts and myoblasts have a very low GAD activity that is not stimulated after transformation by RSV. Neuronal clones, previously derived from ts NY-68-transformed established NR cell lines, have a high GAD activity which is regulated by pp60v-src, while other clones have a low GAD activity apparently not regulated by pp60v-src. These data indicate that pp60v-src selectively activates the expression of GAD in distinct neuronal cells of quail embryo NR cultures transformed by RSV. GAD activity is also stimulated in NR cells infected with viruses containing v-mil. PMID- 2992934 TI - Secondary structure of a channel-forming protein: porin from E. coli outer membranes. AB - Porin from Escherichia coli outer membranes has been analysed by high angle diffuse X-ray diffraction, and by attenuated total reflection infrared spectroscopy. These methods demonstrate independently that the majority of the polypeptide backbone is arranged in anti-parallel beta-pleated sheet structure. The average length of the beta-strands, which are oriented nearly normal to the membrane plane, is estimated to be 10-12 residues, independent of the method used. Although the details of strand arrangement (beta-barrels or stacked sheets) are not as yet known, porin represents the first transmembrane protein for which beta-structure has been established unequivocally. PMID- 2992935 TI - Folding patterns of porin and bacteriorhodopsin. AB - Porin spans the outer membrane of Escherichia coli with most of the protein embedded within the membrane. It lacks pronounced hydrophobic domains and consists predominantly of beta-pleated sheet. These observations require the accommodation of polar and ionizable residues in an environment that has a low dielectric constant. Owing to a currently limited understanding of the constraints governing membrane protein structure, a minimal approach to structure prediction is proposed that identifies segments causing polypeptides to reverse their direction (turn identification). The application of this procedure avoids hydrophobicity parameters and yields a model of porin which is in good agreement with all experimental data available. The presence of polar and ionizable residues within membrane boundaries implies a dense (saturating) network of hydrogen bond donor and acceptor groups. Application to a paradigm of hydrophobic membrane proteins, bacteriorhodopsin, reveals a pattern consistent with its alpha helical folding. The postulated structure includes significantly more polar residues in the membrane domain than have been assumed previously, suggesting that there are also hydrogen bonding networks in bacteriorhodopsin. Extensive networks permeating protein interior and surfaces would explain the extraordinary stability and the tight interactions between functional units in the formation of crystalline arrays of both proteins. PMID- 2992936 TI - Identification and mutational analysis of the promoter for a spinach chloroplast transfer RNA gene. AB - A transcription extract from purified spinach chloroplast was used to test chloroplast DNA sequences for their function as promoter elements. Chloroplast tRNA genes are correctly transcribed in the extract by a soluble RNA polymerase, and precursor molecules are processed into mature tRNAs. Transcription of the spinach chloroplast tRNA2Met gene (trnM2) in vitro requires 5' upstream DNA sequences. Deletion of 5' DNA sequences with exonuclease Bal31 was used to establish the 5' boundary of the promoter region. This boundary is part of a DNA sequence with partial homology to the prokaryotic -35 region. Seventeen base pairs downstream from this sequence a DNA sequence occurs which is homologous to the prokaryotic -10 region. We used synthetic oligonucleotides fused to trnM2 5' deletion mutants to create insertions, deletions and base substitutions in these regions. Internal deletion mutants demonstrated that the -10 promoter element is also required for transcription in vitro. The arrangement of DNA sequences recognised by the chloroplast RNA polymerase resembles the prokaryotic promoter organization. PMID- 2992937 TI - Erythroid-specific expression of human beta-globin genes in transgenic mice. AB - Transgenic mice carrying human beta-globin genes were produced by microinjecting linear DNA molecules containing cloned beta-globin genes with up to 4300 bp of 5' flanking sequence and 1700 bp of 3'-flanking sequence. Most (15 of 20) of these transgenic mice expressed the human beta-globin genes in blood cells and the level of expression in some mice was comparable with that obtained from endogenous beta-globin genes. Human beta-globin gene expression appeared to be restricted to cells of the erythroid lineage and was first detected between 11 and 14 days of development, in parallel with mouse beta-globin. Constructs with as little as 48 bp of 5'-flanking sequence also appeared to be expressed appropriately. The mRNA transcripts had correct 5' ends and directed human beta globin synthesis in reticulocyte lysates. Human beta-globin protein was detectable in mature erythrocytes from progeny of one of these mice. The frequency and extent of expression was severely depressed when the procaryotic vector DNA was not removed prior to microinjection. PMID- 2992938 TI - Detection of a high frequency RsaI polymorphism in the human pro alpha 2(I) collagen gene which is linked to an autosomal dominant form of osteogenesis imperfecta. AB - Screening of the pro alpha 2(I) collagen genes of Southern African populations for restriction fragment length polymorphisms (RFLPs) has revealed a locus polymorphic for the restriction enzyme RsaI. The frequency of the RFLP was 0.38 in Afrikaners, but much lower in indigenous Southern African populations, which suggests that it is of European origin. The polymorphism was used to study 19 affected and non-affected individuals in a four generation family with the autosomal dominant disorder, osteogenesis imperfecta (OI) type I. Co-inheritance of the loss of the RsaI site and the OI phenotype was observed with a lod score of 3.91 at a recombination fraction (theta) of zero, indicating strong linkage. This suggests that the defect in this family is caused by a structural mutation within or close to the pro alpha 2(I) collagen gene. The use of this high frequency RFLP together with other recently described polymorphisms at this locus will facilitate the analysis of the role of this gene in OI and other inherited disorders of connective tissue. PMID- 2992940 TI - Specific transforming potential of oncogenes encoding protein-tyrosine kinases. AB - Several chimeric murine retroviruses were constructed to test whether the gag sequence of Abelson murine leukemia virus (A-MuLV) could influence the in vitro specificity of two sarcoma-inducing oncogenes: src of Rous sarcoma virus and fps of Fujinami sarcoma virus. Although the src- or fps- containing chimerae could transform fibroblasts, they were unable to mimic the action of A-MuLV in causing lymphoid transformation in vitro. A-MuLV-derived gag sequences could, however, functionally replace the 5' end of src and restore the transformation potential of a 5'-truncated src gene. To investigate this functional similarity, we replaced the gag sequence of an A-MuLV virus with the 5' end of src. This recombinant virus behaved like the A-MuLV virus from which it was derived: it transformed both fibroblasts and lymphoid cells in vitro. Taken together, these results suggest that lymphoid transformation in vitro is a specific property of abl and not of src or fps. Furthermore, it shows that a functional homology exists between the gag sequence of A-MuLV and the 5' end of src. PMID- 2992939 TI - Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. AB - Cellular and plasma fibronectins are heterodimers consisting of similar but not identical polypeptides. The differences between fibronectin subunits are due in part to the variability of internal primary sequences. This results from alternative splicing in at least two regions (ED and IIICS) of the pre-mRNA. The complete primary structure of human fibronectin, including most of the internal variations, has been determined by sequencing a series of overlapping cDNA clones. In total, they covered 7692 nucleotides and represented the mRNA sequence coding from the amino terminus of the mature protein to the poly(A) tail. The deduced amino acid sequence of fibronectin has been analysed in terms of the arrangement of internal homologies and the different binding domains. PMID- 2992941 TI - The v-sis/PDGF-2 transforming gene product localizes to cell membranes but is not a secretory protein. AB - The v-sis transforming gene encodes the woolly monkey homologue of human platelet derived growth factor (PDGF) polypeptide 2. After its synthesis on membrane bound polyribosomes, the glycosylated precursor dimerizes in the endoplasmic reticulum and travels through the Golgi apparatus. At the cell periphery, the precursor is processed to yield a dimer structurally analogous to biologically active PDGF. Small amounts of two incompletely processed forms are detectable in tissue culture fluids of simian sarcoma virus (SSV) transformants. However, the vast majority remains cell associated. Thus, this growth factor-related transforming gene product is not a classical secretory protein. These findings define possible cellular locations where the transforming activity of the sis-PDGF-2 protein may be exerted. PMID- 2992942 TI - Proviral activation of the putative oncogene Pim-1 in MuLV induced T-cell lymphomas. AB - Proviral integration near the Pim-1 gene is frequently observed in murine leukemia virus induced T-cell lymphomas in mice. Integration in the Pim-1 domain is associated with the presence of enhanced levels of a Pim-1 mRNA, which is normally expressed as a predominant 2.8 kb species at low levels in lymphoid tissues. The majority of integrations occurred in the 3' region of the Pim-1 transcription unit. This resulted in transcripts ranging in size from 2.0 to 2.6 kb, which were terminated in the 5' proviral LTR. Dependent on the site of integration up to 1300 bases of Pim-1 specific sequences were missing from the modified Pim-1 mRNA in these lymphomas. PMID- 2992943 TI - Common features of polyomavirus mutants selected on PCC4 embryonal carcinoma cells. AB - The genomic rearrangements of six polyomavirus mutants selected on PCC4 embryonal carcinoma cells have been compared and their common characteristics pointed out. All mutants show a duplication which includes at least the adenovirus type 5 (Ad5) E1A-like enhancer core sequence plus a deletion of variable size and location. The presence of the second enhancer core sequence, the SV40-like enhancer, is not required for expression of the PyEC PCC4 phenotype. Two of these mutants are also able to express polyomavirus T antigen on F9 and LT1 cells. Multiadaptation seems to require the duplication of the Ad5 E1A-like core sequence, the maintenance of the SV40-like core sequence and a local change in DNA stability. PMID- 2992944 TI - Identification of the coding region for a second Epstein-Barr virus nuclear antigen (EBNA 2) by transfection of cloned DNA fragments. AB - Cell lines were established by co-transfection of cloned M-ABA Epstein-Barr virus (EBV) DNA fragments with plasmids conferring resistance to dominant selective markers. A baby hamster kidney cell line carrying the HindIII-I1 fragment exhibits a nuclear antigen of 82 000 daltons, serologically defined as EBV determined nuclear antigen (EBNA) 1. Furthermore, a Rat-1 cell line transfected with DNA of the clone pM 780-28 containing three large internal repeats (BglII-U) and the adjacent BglII-C fragment expresses a nuclear antigen of 82 000 daltons which can be visualized only by a subset of anti EBNA-positive human sera. Sera recognizing the 82 000-dalton protein of the transfected cell line reacted with a protein of the same size in the non-producer line Raji, designated as EBNA 2. Conversely, sera without reactivity to the 82 000-dalton protein failed to react with EBNA 2 of Raji cells. P3HR-1 and Daudi cells with large deletions in BglII-U and -C are devoid of EBNA 2. The data presented provide evidence that a second EBNA protein is encoded by the region of the EBV genome which is deleted in the non-transforming P3HR-1 strain. PMID- 2992945 TI - Antibodies against synthetic peptides react with the second Epstein-Barr virus associated nuclear antigen. AB - Five peptides were synthesized on the basis of amino acid sequences predicted from the transformation-associated BamHI WYH region of the genome of the Epstein Barr virus (EBV). Antisera to two peptides deduced from a 1.6-kb open reading frame in the BamHI H fragment identified an 87 000-dalton nuclear polypeptide that was present in EBV-carrying cell lines that expressed the second EBV determined nuclear antigen (EBNA-2). This polypeptide was not detected in cell lines that carried EBV variants with a deleted BamHI WYH region or in EBV negative cell lines. Three peptides deduced from the 1.6-kb open reading frame reacted with human EBNA-positive sera, but not with EBNA-negative sera. Following affinity purification with the peptides, two of the corresponding human antibodies also reacted with the 87 000-dalton polypeptide. PMID- 2992946 TI - The presence of bovine papillomavirus type 4 DNA is not required for the progression to, or the maintenance of, the malignant state in cancers of the alimentary canal in cattle. AB - In the Western Highlands of Scotland there is a very high incidence of alimentary cancers in cattle. The carcinomas of the upper alimentary canal are found in association with virus-induced benign papillomas, and transformation of papillomas to carcinomas has been observed. Strong circumstantial evidence suggests that the progression to malignancy is due to the interplay between the virus, bovine papillomavirus type 4 (BPV-4), and carcinogen(s) present in bracken fern, which infests the marginal upland grazing grounds. The carcinomas are often accompanied by adenomas and adenocarcinomas of the lower bowels. To elucidate the role of the virus in the transformation process, we have analysed several malignancies of the alimentary canal, and have detected the viral genome in only one case of transforming papilloma of the oesophagus and one case of carcinoma of the tongue. We conclude that, although required for the induction of papillomas, the presence of the BPV-4 DNA is not necessary for the progression to, or the maintenance of, the transformed state. PMID- 2992947 TI - Point mutational analysis of the Xenopus laevis 5S gene promoter. AB - We have introduced C to T transitions into GC and CG base pairs of the Xenopus laevis somatic 5S gene coding region and its 5' flank in order to analyse their effects on transcription activity and regulation. These studies allow us to differentiate between the two promoter elements and their spacer, within the internal control region. Mutations within the 5' element which dissent from the corresponding tRNA consensus sequence reduce transcription activity substantially without significantly affecting transcription factor (TF) III A binding. Mutations in the spacer region have no pronounced effect on transcription. The 3' promoter element is found to extend to position 97, since mutations in this region interfere with transcription activity. This may be, at least partially, attributable to a reduced competition strength for TF III A. PMID- 2992948 TI - Endocytosis in Saccharomyces cerevisiae: internalization of enveloped viruses into spheroplasts. AB - When vesicular stomatitis virus was incubated with Saccharomyces cerevisiae spheroplasts at 37 degrees C, part of the virus was internalized by the spheroplasts as shown by the following criteria. (i) The spheroplast-associated virus was protected from proteinase K digestion, which releases surface-bound virus by degrading the envelope glycoproteins. (ii) The spheroplast-associated virus was resistant to mild Triton X-100 treatment, which readily solubilizes the virus. The same results were obtained with Semliki Forest virus. Internalization of the two viruses followed linear kinetics up to 90 min at 37 degrees C. Internalization was concentration- and temperature-dependent. At 11 degrees C no uptake could be detected for at least 2 h. Homogenization and organelle fractionation protocols were designed for the S. cerevisiae spheroplasts to study the compartments into which the virions were internalized. Three compartments containing both marker viruses could be separated in density gradients. One coincided with vacuole markers, one banded at a slightly higher and one at a similar density to the plasma membrane markers. Thus, S. cerevisiae spheroplasts appear to have the capability of endocytosing particulate markers like viruses. The companion paper describes internalization of two soluble macromolecules, alpha-amylase and fluorescent dextran, into intact cells. PMID- 2992949 TI - Role of conserved sequence elements in yeast centromere DNA. AB - Conserved sequence features in Saccharomyces cerevisiae CEN DNA are confined to a region of approximately 120 bp. The highly conserved 8 bp at the left (PuTCACPuTG) constitute the left boundary of a functional CEN DNA as shown by the analysis of a series of Bal31 deletions. The right boundary of a functional CEN DNA lies within the conserved 25 bp at the right (TGT-T-TG--TTCCGAA-----AAA) or a few base pairs further outside of the 120-bp region. One mutant which just lacks the left conserved DNA element PuTCACPuTG can still assemble into a partially functional mitotic centromere and it assembles into a well functioning meiotic centromere. The sequences between the two conserved terminal DNA elements can be increased in length (+50%) or in GC content (from 6% to 12%) without measurable changes in mitotic and meiotic segregations of plasmids carrying such CEN mutations. The naturally occurring length and GC content of this centromere DNA sequence element is, therefore, not essential for centromere function. We discuss the possibility that it partly acts as a hinge region between two domains. Finally, we tested integrations of CEN DNA into the genome and found a toleration of wild-type CEN6 DNA when present 3' of the LYS2 gene. PMID- 2992950 TI - Hairpin plasmid--a novel linear DNA of perfect hairpin structure. AB - The terminal structures of deletion derivatives of linear DNA killer plasmid from yeast were analyzed. The yeast Kluyveromyces lactis harbors two unique double stranded linear DNA killer plasmids, pGKL1 of 8.9 kb and pGKL2 of 13.4 kb. The killer toxin and the resistance to the killer are coded by pGKL1, while pGKL2 is required for the maintenance of pGKL1 in the cell. When the pGKL plasmids from K. lactis were transferred into Saccharomyces cerevisiae by transformation, non killer transformants harboring pGKL2 and new plasmids, F1 of 7.8 kb and F2 of 3.9 kb, were obtained. F2 was shown to be a linear DNA arising from a 5-kb deletion of the right part of pGKL1. F1 was an inverted dimer of F2. Here we show that F2 has two different terminal structures: one end has a protein attached at the 5' terminus whereas the two strands of duplex are linked together at the other end, thus forming a hairpin structure. This is a novel type of autonomously replicating DNA molecule. PMID- 2992951 TI - Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination. AB - The mature 3' end of Escherichia coli tryptophan operon mRNA in vivo coincides with a site (trp t) having features commonly associated with rho-independent terminators in bacteria. Efficient generation of this 3' end in vivo is nevertheless affected by a distal rho-dependent site (trP-t'), though these two sites behave independently in vitro. We have cloned these sites upstream of the galactokinase gene (galK), and galactokinase levels in vivo indicate that, as terminators per se, their efficiencies (37% for trp t, and 79% for trp t') do not differ significantly from those observed in vitro. However, when the trp t hairpin is placed between galK and a downstream copy of trp t', galactokinase levels are enhanced 2- to 3-fold. This suggests the involvement of a post transcriptional event, such as RNA processing, in determining the level of gene activity. Indeed, in the presence of the 3' exonuclease RNase II, mRNA terminated by rho factor in vitro at the trp t' site is processed back to the trp t site. The remote trp t' region appears to be the major termination site for trp mRNA, and the trp t hairpin serves a dual function-as a minor terminator, and as a protective barrier to 3' exonucleolytic degradation. We infer that the tandem terminators, rho factor, and RNA processing are all required to generate the mature 3' end of this bacterial mRNA. PMID- 2992952 TI - Resistance, regulatory and production genes for the antibiotic methylenomycin are clustered. AB - At least 17 kb of DNA from the large unisolatable Streptomyces coelicolor A3(2) plasmid SCP1 are concerned with methylenomycin biosynthesis. Mutational cloning analysis, using insert-directed integration of att site deleted phage vectors into an SCP1-containing host, provided evidence of two large transcription units, of at least 6.6 kb and 9.5 kb. At the leftmost apparent end of the larger (left hand) transcription unit is a region apparently involved in negative regulation of methylenomycin biosynthesis: when fragments from this region were used to direct phage integration, marked overproduction of methylenomycin resulted. The methylenomycin resistance determinant is located at the rightmost end of this same transcription unit. Hybridisation analysis with 13 kb of the cloned mmy region showed that it was closely similar to a segment of pSV1, a plasmid that specifies methylenomycin biosynthesis in S. violaceus-ruber SANK 95570. PMID- 2992953 TI - Phosphorylation of the ribosomal protein S6 during agonist-induced exocytosis in exocrine glands is catalyzed by calcium-phospholipid-dependent protein kinase (protein kinase C). Experiments with guinea pig parotid glands. AB - The ribosomal protein S6 in exocrine cells is phosphorylated during stimulation of exocytosis by cAMP-dependent or calcium-dependent agonists. Under both conditions the same tryptic S6 phosphopeptides (termed A, B, and C) were found [Padel, Kruppa, Jahn & Soling (1983) FEBS Lett. 159, 112-118]. Studies have now been made of the phosphorylation pattern of protein S6 from purified guinea pig parotid ribosomes following in vitro phosphorylation with calmodulin-dependent, phospholipid-dependent, and cAMP-dependent protein kinases. Only the phospholipid dependent enzyme led to the phosphorylation of peptides A, B, and C, while the cAMP-dependent enzyme phosphorylated only peptides A and C, and the calmodulin dependent enzyme did not phosphorylate any of the phosphopeptides found in S6 from unstimulated or stimulated intact cells. Guinea pig parotid microsomes contain substantial phospholipid-dependent protein kinase activity. Stimulation of intact parotid glands with tetradecanoylphorbol acetate led to a significant phosphorylation of S6 and a similar tryptic S6 phosphopeptide pattern as seen with carbamoylcholine. It is concluded that activation of phospholipid-dependent protein kinase is responsible for the phosphorylation of protein S6 during stimulation with calcium-dependent and cAMP-dependent secretagogues. PMID- 2992954 TI - The structure of the double-stranded RNA pentamer 5'(CACAG) . 5'(CUGUG) determined by nuclear Overhauser enhancement measurements: interproton distance determination and structure refinement on the basis of X-ray coordinates. AB - The structure in solution of the duplex RNA pentamer 5'(CACAG) . 5'(CUGUG), comprising the stem of the T psi C loop of yeast tRNAPhe, has been investigated by means of one- and two-dimensional nuclear Overhauser enhancement measurements. All non-exchangeable base and sugar proton resonances with the exception of the H5'/H5" sugar resonances are assigned in a sequential manner. From the relative intensities of the cross-peaks obtained in the pure-phase absorption two dimensional nuclear Overhauser enhancement spectra at several mixing times, it is deduced that the RNA pentamer adopts an A-type conformation in solution. Cross relaxation rates and interproton distances are determined from the time dependence of the nuclear Overhauser effects, principally by one-dimensional measurements. The structure of the RNA pentamer is then refined by restrained least-squares minimization on the basis of both distance and planarity restraints using fibre diffraction data as an initial model. The refined structure of the RNA pentamer is of the A type but exhibits local structural variations in glycosidic bond and backbone torsion angles as well as in propeller twist, base roll and base tilt angles. PMID- 2992955 TI - Amino acid sequence of histone H1 at the ADP-ribose-accepting site and ADP-ribose X histone-H1 adduct as an inhibitor of cyclic-AMP-dependent phosphorylation. AB - The ADP-ribosylation site of histone H1 from calf thymus by purified hen liver nuclear ADP-ribosyltransferase was determined and effects of the ADP-ribose X histone-H1 adduct on cAMP-dependent phosphorylation of the histone H1 were investigated. ADP-ribosylated histone H1 was prepared by incubation of histone H1, 1 mM [adenylate-32P]NAD and the purified ADP-ribosyltransferase. N Bromosuccinimide-directed bisection of ADP-ribosylated histone H1 showed that the NH2-terminal fragment (Mr = 6000) was modified and contained serine residue 38, the site of phosphorylation by cAMP-dependent protein kinase. Digestion of the NH2-terminal fragment with cathepsin D and trypsin, and purification of this fragment, using high-performance liquid chromatography, yielded a radiolabelled single peptide corresponding to residues 29-34 of histone H1, containing the arginine residue as the ADP-ribosylation site. These results indicate that ADP ribosylation of histone H1 occurs at the arginine residue 34, sequenced at the NH2-terminal side of the phosphate-accepting serine residue 38. Phosphorylation of histone H1 from calf thymus by cAMP-dependent protein kinase was markedly reduced when histone H1 was ADP-ribosylated. Kinetic studies of phosphorylation revealed that ADP-ribosylated histone H1 was a linear competitive inhibitor of histone H1 and a linear non-competitive inhibitor of ATP. PMID- 2992956 TI - Cyclic nucleotide specificity of the activator and catalytic sites of a cGMP stimulated cGMP phosphodiesterase from Dictyostelium discoideum. AB - The cellular slime mold Dictyostelium discoideum has an intracellular phosphodiesterase which specifically hydrolyzes cGMP. The enzyme is activated by low cGMP concentrations, and is involved in the reduction of chemoattractant mediated elevations of cGMP levels. The interaction of 20 cGMP derivatives with the activator site and with the catalytic site of the enzyme has been investigated. Binding of cGMP to the activator site is strongly reduced (more than 80-fold) if cGMP is no longer able to form a hydrogen bond at N2H2 or O2'H. Modifications at N7, C8, O3' and O5' induce only a small reduction of binding affinity. A cyclic phosphate structure, as well as a negatively charged oxygen atom at phosphorus, are essential to obtain activation of the enzyme. Substitution of the axial exocyclic oxygen atom by sulphur is tolerated; modification of the equatorial oxygen atom reduces the binding activity of cGMP to the activator site by 90-fold. Binding of cGMP to the catalytic site is strongly reduced if cGMP is modified at N1H, C6O, C8 and O3', while modifications at N2H2, N3, N7, O2'H, and O5' have minor effects. Both exocyclic oxygen atoms are important to obtain binding of cGMP to the catalytic site. The results indicate that activation of the enzyme by cGMP and hydrolysis of cGMP occur at different sites of the enzyme. cGMP is recognized at these sites by different types of molecular interaction between cGMP and the protein. cGMP derivatives at concentrations which saturate the activator site do not induce the same degree of activation of the enzyme (activation 2.3-6.6-fold). The binding affinities of the analogues for the activator site and their maximal activation are not correlated. Our results suggest that the enzyme is activated because cGMP bound to the activator site stabilizes a state of the enzyme which has a higher affinity for cGMP at the catalytic site. PMID- 2992957 TI - Electrostatic effects and calcium ion concentration as modulators of acid phosphatase bound to plant cell walls. AB - At 'low' ionic strength, acid phosphatase bound to plant cell walls exhibits an apparent negative co-operativity, whereas it displays classic Michaelis-Menten kinetics in free solution. Conversely, at 'high' ionic strength, the bound enzyme and the soluble enzyme behave identically. This apparent negative co-operativity is explained by the existence of an electrostatic partition of the charged substrate by the fixed negative charges of the cell wall. Raising the ionic strength suppresses these electrostatic repulsion effects. Calcium may be removed from the cell walls by acid treatment and the acid phosphatase is apparently strongly inhibited. This inhibition occurs together with an increased apparent negative co-operativity of the enzyme. Incubating cell wall fragments previously depleted of calcium with CaCl2 restores the initial behaviour of the enzyme. Calcium, which tightly binds to cell wall pectic compounds, has by itself no effect on the enzyme in free solution. It affects the net charge of the cell wall and therefore the amplitude of electrostatic repulsion effects. Non-linear least square fitting methods make it possible to estimate the density of fixed negative charges as well as the electrostatic partition coefficient, for both the 'native' and 'calcium-deprived' cell wall fragments. It may be shown directly that calcium loading and unloading in the cell wall controls the electrostatic effects, by monitoring proton extrusion from cell wall fragments upon raising the ionic strength. Proton outflux in the bulk phase is considerably enhanced upon removal of calcium from the cell walls. The main conclusion is that loading and unloading of calcium during cell elongation and division may regulate the activity of cell wall enzymes. PMID- 2992958 TI - Inhibition, by lanthanides, of neutral proteinases secreted by human, rheumatoid synovium. AB - Fragments of human, rheumatoid synovium were maintained on organ culture for three days under serum-less conditions. Their conditioned media contained collagenolytic, gelatinolytic and caseinolytic activities, which were susceptible to inhibition by lanthanide ions. Of the four lanthanides tested, Sm3+ proved the best inhibitor of gelatinase and caseinase, while La3+ inhibited collagenase the most strongly. Inhibition of collagenase by La3+ was uncompetitive. A direct binding assay confirmed the greater association between collagen fibrils and collagenase in the presence of La3+. Ca2+ was not required for binding of the uninhibited enzyme to collagen, but acted to stabilize collagenase against thermoinactivation. PMID- 2992959 TI - Nucleotide sequence of the pyrD gene of Escherichia coli and characterization of the flavoprotein dihydroorotate dehydrogenase. AB - Dihydroorotate dehydrogenase (EC 1.3.3.1) was purified to near electrophoretic homogeneity from the membranes of a strain of Escherichia coli carrying the pyrD gene on a multicopy plasmid. The preparation had a specific activity of 120 mumol min-1 mg-1 and contained flavin mononucleotide (FMN) in amounts stoichiometric to the dihydroorotate dehydrogenase subunit (Mr = 37000). The flavin group was reduced when dihydroorotate was added in the absence of electron acceptors. The complete sequence of 1357 base pairs of an EcoRI-EcoRI DNA fragment containing the pyrD gene was established. Dihydroorotate dehydrogenase is encoded by a 336 triplets open reading frame. The molecular mass (Mr = 36732), the amino acid composition and the N-terminal sequence of the predicted polypeptide agree well with the data obtained by analysis of the purified protein. A region of the amino acid sequence (residues 292-303, i.e. Ile-Ile-Gly-Val-Gly-Gly-Ile-Asp-Ser-Val-Ile Ala) shows distinct homology to the cofactor binding site of other flavoproteins. No hydrophobic regions large enough to span the cytoplasmic membrane were observed. By the S1-nuclease technique an mRNA start was mapped 34 +/- 2 nucleotide residues upstream of the beginning of the coding frame of pyrD. The leader region contains no similarity to the attenuators of the pyrB and pyrE genes of E. coli. PMID- 2992960 TI - The mechanism of ion selectivity of OmpF-porin pores of Escherichia coli. AB - The OmpF porin from the outer membrane of Escherichia coli acts as a lightly cation-selective pore, allowing the diffusion of small cations and cationic molecules, whose Mr are a little larger than the threshold exclusion limit. To ascertain the mechanism of this cation selectivity, we have examined a possible influence of cationic solutes on the fluorescence emission and the circular dichroic spectrum of tryptophan residues of the porin trimer, searching for conformational change(s). The diffusion of cationic solutes was determined with the native and the amidated porins in the presence or the absence of the effector cations. The following results were obtained. (a) Cations, e.g. spermidine, caused fluorescence quenching in the native trimer, with a half-maximum fluorescence quenching at 11-18 microM. A change in the circular dichroic spectrum was also recorded at around 280 nm. (b) The dissociation constant of spermidine to the native trimer was calculated to be 16 microM as determined by the method of equilibrium dialysis. (c) The cation-caused fluorescence quenching was reversed when the carboxyl groups of the trimer were modified by the amidation reaction, though amidation of the trimer resulted in no significant change in the fluorescence intensity. (d) The diffusion rate of N benzyloxycarbonyl-glycyl-L-prolyl-L-arginine p-nitroanilide through the native and the amidated porins was lowered in the presence and the absence, respectively, of cations. Both the extent of fluorescence quenching in the presence of cation and the rate of cation diffusion were inversely proportional to the number of amidated carboxyl residues. The relative fluorescence quenching of the porin trimer (the amidated versus the native) in the presence of cations was linearly related to the relative solute diffusion via the porin (the amidated versus the native). These results suggested that cations caused a conformational change in the trimer, resulting in an easier diffusion of the solutes. The results suggested further that a limited number of carboxyl groups in the pore interior are involved in the cation selectivity of OmpF-porin pores. PMID- 2992961 TI - Systematic application of high-resolution, phase-sensitive two-dimensional 1H-NMR techniques for the identification of the amino-acid-proton spin systems in proteins. Rabbit metallothionein-2. AB - Novel strategies for elucidation and classification of amino acid 1H-NMR spin systems in proteins were developed exploiting recently introduced two-dimensional NMR techniques such as phase-sensitive double-quantum-filtered correlated spectroscopy, relayed coherence transfer spectroscopy, double quantum spectroscopy and nuclear Overhauser spectroscopy. Due to the improved resolution in phase-sensitive spectra, the fine structure of cross peaks could be exploited as a powerful source of information for establishing 1H-1H connectivities. Principles for the interpretation of multiplet structures of absorption mode cross peaks are discussed. With these methods the 1H spin systems of rabbit liver metallothionein-2 were elucidated and classified according to amino acid types. Despite the intrinsically difficult situation arising from the unusual amino acid composition of this protein, a more complete characterization of the 1H spin systems prior to the step of sequential resonance assignments was achieved with the presently introduced methodology than was possible in earlier studies of proteins of similar size. PMID- 2992962 TI - Antiviral activity in L1210 cells of liposome-encapsulated (2' 5')oligo(adenylate) analogues. AB - Evidence is available for a role of a (2'-5')(A)n-activated endoribonuclease (RNase L) in the antiviral activity of interferon for several RNA viruses. (2' 5')(A)n and their analogues might thus provide an interesting alternative to exogenous interferons or their inducers in antiviral chemotherapy. In addition, the evaluation of the activity of (2'-5)(A)n as mediators of interferon's biological activities or as cell growth regulators requires biochemical studies using agonists or antagonists of the system. Non-disruptive techniques for the introduction of (2'-5')(A)n and their analogues into cell lines or tissues are required for these studies since these highly charged compounds are cell impermeable. (2'-5')(A)n oligomers and analogues of increased stability towards phosphodiesterases were derived by chemical modification of their 2' end and encapsulated in protein-A-bearing liposomes. The specific delivery of liposome contents into L1210 mouse leukemic cells was achieved with the help of monoclonal antibodies directed against the appropriate class I major histocompatibility complex-encoded proteins expressed by these cells. This intracellular delivery led to transient inhibition of protein synthesis and an antiviral activity, both compatible with activation of RNase L. This activity was enhanced for the analogues designed to resist degradation, with respect to the natural product. PMID- 2992963 TI - Adenosine kinase from bovine adrenal medulla. AB - Adenosine kinase from bovine adrenal medulla was purified 1600-fold by using ammonium sulfate precipitation, gel filtration and affinity chromatography. Gel filtration yielded a relative molecular mass around 42000 and Michaelis constants were 0.2 microM for adenosine and 20 microM for MgATP. The enzyme showed a broad specificity for purine nucleoside triphosphate as phosphate donors. Both free Mg2+ and ATP were inhibitors. AMP was a competitive inhibitor with regard to adenosine and a non-competitive inhibitor versus MgATP, while ADP was a uncompetitive inhibitor with regard to adenosine and a non-competitive inhibitor versus MgATP. Adenosine kinase was strongly inhibited by the bis(adenylyl) polyphosphates Ap4A and Ap5A. These compounds inhibited the enzyme competitively versus MgATP (Ki = 0.06 microM for Ap4A and 0.4 microM for Ap5A) and uncompetitively with regard to adenosine. The results of the kinetic analysis suggest an ordered bi-bi mechanism, adenosine being the first substrate. The phosphorylation of adenosine was unaffected in the presence of vanadate ions. PMID- 2992965 TI - An improved procedure for purifying 5'-nucleotidase from various sources. Evidence for tissue and species differences in their molecular mass and affinity for F-actin. AB - 5'-Nucleotidase from chicken gizzard smooth muscle has been extracted, using a sulfobetaine derivate of cholic acid, and purified to homogeneity by employing three chromatographic steps. It is shown that the purification scheme can be applied to 5'-nucleotidase from other sources, such as rat liver. On sodium dodecyl sulfate polyacrylamide gels, stained with silver nitrate, the purified enzyme from chicken gizzard shows a single polypeptide band with an apparent molecular mass of 79 kDa. The enzyme purified from rat liver exhibits a molecular mass of 73 kDa in agreement with published data [Bailyes, E.M., Soos, M., Jackson, P., Newby, A. C., Siddle, K. & Luzio, J.P. (1984) Biochem. J. 221, 369 377). Gel filtration, using non-denaturating detergent solutions, indicates that the native enzyme may exist as a homodimer (152 kDa) or homotetramer (310 kDa). Antibodies raised against the enzyme purified from chicken gizzard bind only 5' nucleotidase, solubilized from chicken muscular sources, when immobilized, but not from chicken or rat liver. The existence of tissue specific variants of 5' nucleotidase is therefore postulated and it appears that these particular isoforms can also be classified in membranous and secretory forms of 5' nucleotidase. They also differ in their mode of interaction with actin. The AMPase activity of the membranous (= muscular) isoform is inhibited to a considerably higher percentage by F-actin than the enzyme isolated from rat liver. PMID- 2992964 TI - Investigation of the preferred Mg(II)-adenine-nucleotide complex at the active site of ectonucleotidases in intact vascular cells using phosphorothioate analogues of ADP and ATP. AB - In the presence of Mg2+ the ecto-(nucleoside diphosphatase) on intact vascular endothelial or smooth muscle cells in culture selectively catabolizes the PS diastereoisomer of adenosine 5'-[alpha-thio]diphosphate, (PS)-ADP [alpha S], and the ecto-(nucleoside triphosphatase) selectively catabolizes the PS isomer of adenosine 5'-[beta-thio]triphosphate, (PR)-ATP[beta S], but exhibits no selectivity towards ATP[alpha S] isomers. In the presence of Cd2+ selectivity to ADP[alpha S] and to ATP[beta S] isomers is reversed; in the presence of Co2+, selectivity is lost. We conclude that each enzyme preferentially recognises the lambda (screw-sense) bidentate Mg(II)-nucleotide complex at its active site. PMID- 2992966 TI - Covalent cross-linking of vasoactive intestinal peptide (VIP) to its receptor in intact colonic adenocarcinoma cells in culture (HT 29). AB - [125I]Monoiodinated vasoactive intestinal peptide (125I-VIP) was cross-linked with human colonic adenocarcinoma cells (HT29 cells) grown as a monolayer using dithiobis(succinimidylpropionate) as cross-linking reagent. The cross-linked polypeptides were separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. A major polypeptide of Mr = 67 000 was characterized and it behaved like a high-affinity binding site for VIP according to the following data. The concentration of native VIP (0.5 nM) giving half-maximum inhibition of 125I-VIP covalent cross-linking with this polypeptide was very similar to that giving half-maximum displacement of 125I-VIP on HT 29 cells (0.6 nM). Glucagon or insulin was unable to inhibit the labelling of the Mr-67 000 component. In our experimental conditions neither specific 125I-VIP binding nor covalent labelling was observed with monolayers of Madin Darby canine kidney epithelial cells (MDCK cells) or African green monkey kidney fibroblasts (Vero cells) while the Mr-67 000 polypeptide was also characterized with human rectal adenocarcinoma cells (HRT 18 cells), known to possess the VIP receptor. Preincubation of HT 29 cells with native VIP at 37 degrees C, before 125I-VIP binding and subsequent cross-linking reaction, decreased the labelling of the Mr 67 000 polypeptide up to 80%. Assuming one molecule of 125I-VIP cross-linked per polypeptide, we have characterized, for the first time, a major polypeptide of Mr = 64 000, which belongs to the high-affinity VIP binding site of an intestinal human cell line. PMID- 2992967 TI - The role of protein phosphorylation in the hormonal control of enzyme activity. PMID- 2992969 TI - The domain structure of tryptophan synthase. A neutron scattering study. AB - Tryptophan synthase from Escherichia coli is a complex of two alpha subunits and two beta subunits. Small-angle neutron scattering involving deuterium-labelled isomers revealed the quaternary structure of the enzyme at the level of the beta 2 subunit and the two structural domains P1 and P2 which constitute the alpha subunits. Within the alpha 2 beta 2 complex, the two alpha subunits are completely separated. They are situated on opposite sides of the beta 2 subunit. The most probable distance between the two alpha protomers is 10.5 +/- 1 nm; the nearest distance is 5.8 +/- 0.5 nm, and the largest distance is 13.5 +/- 0.5 nm. The two domains of the same alpha subunit are intimately juxtaposed. The distances between two like or unlike domains belonging to opposite alpha subunits are roughly equal. All domains exhibit about equal distances to the beta 2 subunit which is situated in the centre of the complex. Thus the cleft between P1 and P2, which probably contains the active site of the alpha subunit, makes intimate contact with the beta 2 subunit. Neutron scattering allows us to determine the shape of the beta 2 subunit within the complex. Comparison with the free dimer suggests a conformational change, upon assembly, from an elongated into a more compact form. PMID- 2992968 TI - Plant peroxidases. Their primary, secondary and tertiary structures, and relation to cytochrome c peroxidase. AB - The amino acid sequences of the 51% different horseradish peroxidase HRP C and turnip peroxidase TP 7 have previously been completed by us, but the three dimensional structures are unknown. Recently the amino acid sequence and the crystal structure of yeast cytochrome c peroxidase have appeared. The three known apoperoxidases consist of 300 +/- 8 amino acid residues. The sequences have now been aligned and show 18% and 16% identity only, between the yeast peroxidase and plant peroxidase HRP C and TP 7, respectively. We show that different structural tests all support similar protein folds in plant peroxidases and yeast peroxidase and, therefore, a common evolutionary origin. The following tests support this thesis: (a) predicted helices in the plant peroxidases follow the complex pattern observed in the crystal structure of cytochrome c peroxidase; (b) their hydropathic profiles are similar and agree with observed buried and exposed peptide chain in cytochrome c peroxidase; (c) half-cystines which are distant in the amino acid sequence of plant peroxidases become spatial neighbours when fitted into the cytochrome c peroxidase model; (d) the two-domain structure proposed from limited proteolysis of apoperoxidase HRP C is observed in the crystal structure of cytochrome c peroxidase. The similarities and differences of the plant and yeast peroxidases and the reactive side chains of a plant peroxidase active site are described. The characteristics of Ca2+-binding sequences, derived from several superfamilies, are applied to predict the Ca2+ binding sequences in plant peroxidases. PMID- 2992970 TI - Comparison of the nucleotide sequences of the genes encoding the KS71A and F7(1) fimbrial antigens of uropathogenic Escherichia coli. AB - DNA fragments encompassing the genes for the KS71A and F7(1) fimbrial subunits of Escherichia coli strains KS71 (O4:K12) and AD110 (O6:K2), respectively, have been subjected to DNA sequencing. The nucleotide sequences of the two fimbrillin genes were identical and they encode a polypeptide of 187 amino acids of which 21 amino acids probably will constitute the signal sequence. The primary structure of these fimbrillins showed significant homology with the primary structure of other E. coli fimbrillins. PMID- 2992972 TI - Experimental data on the problem of specific hepatosplenography with radiodense lipomicrons. AB - Investigations into specific hepatosplenography performed with the aid of radiodense lipomicrons of different size ranges and various surface layers are presented. It is concluded that certain synthetic neutral and/or negatively charged amphiphilic substances may enhance hepatic and splenic uptake of small lipomicrons. Kinetics concerning clearance of lipid globules from the blood and hepatic concentration as well as elimination rates are studied. Furthermore, circulatory reactions following the intravenous administration of lipid emulsions and interactions between lipid globules and plasma proteins or synthetic polymeric substances are discussed. PMID- 2992971 TI - Nuclear ADP-ribosyl transferase activity correlates with induction of P-450 monooxygenases by phenobarbital in rat liver microsomes. AB - The effect of phenobarbital treatment on the nuclear ADP-ribosyl transferase activity has been studied in parallel with microsomal cytochrome P-450 concentration and related mono-oxygenase activities, in rat liver. A marked activation of the ADP-ribosyl transferase was observed 24 h after phenobarbital administration. The chronological study performed between 0-6 days after phenobarbital treatment showed a sharp increase in this nuclear enzyme activity, to approximately equal to 270% of the control value produced in 48 h. The administration of 5'-methylnicotinamide in vivo, an inhibitor of ADP-ribosyl transferase activity in vitro, produced a decrease both of the induction of liver microsomal cytochrome P-450 mono-oxygenases and nuclear ADP-ribosyl transferase activity. The role of nuclear ADP-ribosyl transferase in the adaptative response of the liver cell to phenobarbital is discussed. PMID- 2992973 TI - CT diagnosis of abdominal lymph node metastases in hepatocellular carcinoma. AB - CT scanning is useful for diagnosing abdominal lymph node metastasis. Using this technique, histologically confirmed abdominal lymph node metastases were detected in nine of 49 patients (33 autopsy cases and 16 laparotomy cases) with hepatocellular carcinoma (hepatoma). Among the 49 patients, three had periportal (6.1%), six peripancreatic (12.2.%) and six para-aortic adenopathies (12.2%). Two of the patients had adenopathy at all three sites. Retrospectively, CT detected two periportal, four peripancreatic and all six para-aortic adenopathies. Most of the hepatomas with adenopathy showed infiltrative growth; tumour thrombosis of the portal vein was a common complication. PMID- 2992974 TI - Immunoelectron microscopic localization of the restriction endonuclease EcoRI in Escherichia coli BS 5. AB - Purified restriction endonuclease EcoRI isolated from Escherichia coli BS 5 was used for the production of enzyme-specific IgG antibodies in rabbits. For enzyme localization experiments, paraformaldehyde-glutaraldehyde-fixed cells were embedded and polymerized by a low-temperature procedure using Lowicryl K4M. The immuno electron microscopic protein A-gold technique and an immuno-gold method revealed that 70% of the enzyme-specific labeling were located in the cell envelope whereas 30% were found in the cytoplasm. In metal-shadowed preparations, no indications for the presence of the enzyme could be found on the cell surface; however, on the surface of cell protoplasts enzyme specific labeling could be detected. The results indicate the presence of the major amount of EcoRI in the periplasmic space of the cell where it might be loosely bound or even freely diffusible. PMID- 2992975 TI - Isolation of 5'-nucleotidase from chicken gizzard, its properties, and subcellular localization in chicken tissues using monospecific antibodies. AB - After solubilization with zwitterionic detergents 5'-nucleotidase was purified to homogeneity from chicken gizzard. Purified 5'-nucleotidase appeared to be composed of a single polypeptide chain of 79 kDa as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE); gel filtration studies in the presence of detergents, however, indicated that the native enzyme is a homodimer. Antisera against purified chicken gizzard 5'-nucleotidase were raised in rabbits, they were shown to inhibit the enzymic activity of different 5' nucleotidase preparations in a species and tissue specific manner. On frozen sections the antisera stained the cell periphery of chicken gizzard smooth and skeletal muscle and faintly stained cardiac muscle cells when using the indirect immunofluorescent technique. In chicken cardiac tissue, a prominent staining of the vascular system also became apparent. In avian and rat non-muscle tissues (hepatic and pancreatic tissue) the vascular system was always found to be brightly stained, i.e. the vascular smooth muscle and endothelial cells. On frozen sections of chicken liver the sinusoidal region of the hepatocytes was brightly stained, the bile canalicular region, however, only faintly. Using the immunocytochemical technique, a more prominent tissue specificity rather than species specificity of the available antisera became apparent. This may therefore reflect the existence of tissue-specific isoforms of the enzyme. PMID- 2992976 TI - Infection of AtT20 murine pituitary tumour cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region. AB - AtT20 cells, a line of murine pituitary tumour cells that secrete adrenocorticotropic hormone (ACTH), have been infected with the coronavirus mouse hepatitis virus strain A59 (MHV-A59). Between 5% and 10% of AtT20 cells are susceptible to the infection. Unlike infections of fibroblastic sac- and 17Cl 1 cells, the infection of AtT20 cells does not lead to cell fusion, despite the production of the fusogenic E2 viral spike glycoprotein. Within infected AtT20 cells the second viral envelope glycoprotein, E1, is located in a perinuclear region; at least until very late in the infection it fails to accumulate to detectable levels in the rough endoplasmic reticulum (RER). By contrast to infection of sac- and 17Cl 1 cells, where the RER is a major site of assembly of progeny virions, in AtT20 cells budding of progeny virions is restricted to the Golgi cisternae, which eventually vesiculate, and peri-Golgi smooth membraned vesicles. Apparently, therefore, the intracellular compartments into which wild type MHV-A59 buds are determined not by the virus but by the host cells. MHV-A59 infected cultures of AtT20 cells can be serially passaged without loss of the infection or increase in the proportion of infected cells; they become persistently infected carrier cultures. The progeny virus from serially passaged, infected AtT20 cells is apparently wild-type. It infects sac- cells and induces them to form syncitia. Within the sac- syncitia the viral E1 glycoprotein accumulates in the RER and many virions assemble there. PMID- 2992977 TI - Antibody induced internalization of acetylcholine nicotinic receptor: kinetics, mechanism and selectivity. AB - The loss from the cell surface of cholinergic nicotinic receptors induced by exposure to antireceptor antibodies, and the mechanism of this loss has been studied in the BC3H-1 cell line. The maximal effect, i.e. 50% loss of receptors, was observed after 30 min. Receptor loss was due to internalization, which was not accompanied by a detectable increase in fluid phase endocytosis nor by changes in the number and distribution of intramembrane particles (IMP) revealed by freeze-fracture of the plasmalemma. The internalization was specific since the concentration of other receptors exposed at the surface of BC3H-1 cells (alpha and beta adrenergic receptors) was not affected by anticholinergic antibodies. It was temperature-dependent, independent of external calcium, but was blocked by trifluoperazine (TFP), a calmodulin antagonist. The rate of antibodies-induced receptor internalization was much more rapid than that of degradation and furthermore low concentrations of antibodies increased receptor internalization but did not increase receptor degradation. Our findings may be relevant to the clarification of the mechanism of Ab-induced alteration in autoimmune diseases, and in particular in myasthenia gravis. PMID- 2992978 TI - Demonstration of receptor binding of two apo-B containing lipoproteins by differential labelling with colloidal gold. AB - The interaction of two types of apo-B containing lipoproteins, human low density lipoprotein (LDL) and lipoprotein-a (Lp(a], with cultured human skin fibroblasts was studied by differential colloidal gold labelling in conjunction with thin sectioning and surface replication techniques. After separate exposure of the fibroblasts to either gold labelled LDL or Lp(a) for 15 to 30 min at 37 degrees C, labelled lipoproteins were predominantly found in coated pit areas. Excess of unlabelled LDL or Lp(a) completely displaced the gold labelled lipoproteins, indicating specific binding by the LDL-receptor. Simultaneous exposure of fibroblasts to LDL-16 nm gold and Lp(a)-40 nm gold conjugates revealed that both LDL and Lp(a) are bound in the same coated pit and internalized into the same endosome. In contrast to native lipoproteins, gold labelled acetylated lipoproteins were found diffusely distributed on membrane surface areas predominantly representing fibronectin-containing fibrils. PMID- 2992979 TI - Transformation of mammalian cell by iontophoretic pricking or iontophoretic microinjection. AB - Thymidine kinase negative mouse (LATK-) and hamster (BHKTK-) cells were transfected with a recombinant plasmid (pTK1) carrying the thymidine kinase gene of HSV-1 by iontophoretic pricking or iontophoretic microinjection techniques. Transfection frequencies were measured using short term survival and long term transformation assays and were found to be at the level of 10 to 15% and 1 to 2% respectively for mouse or hamster cells transfected with pTK1. The presence of covalently linked transcriptional enhancers from SV40 or Moloney MSV increased long term transformation frequencies approximately 10-fold. Transfection by iontophoretic pricking of non-tumorigenic mouse (NIH3T3) or hamster (BHKC13) cells with a recombinant (pAGT1) carrying the human T24 Ha-ras1 oncogene and the bacterial aminoglycoside phosphotransferase (aph) gene as a selectable marker resulted in transformation of these cells. Southern blot hybridization analyses demonstrated the presence of circular, integrated or rearranged donor DNA molecules in the transformed cells. These gene transfer techniques and iontophoretic pricking in particular should be useful for the transfection of a variety of cell lines and markers. PMID- 2992980 TI - Lysosomal accumulation of cholesterol and sphingomyelin: evidence for inhibition of acid sphingomyelinase. AB - The subcellular site of accumulation of non-esterified cholesterol and sphingomyelin in lipid-laden livers and spleens from rats given multiple intravenous injections with liposomes made up of these lipids were investigated by morphological and biochemical techniques. The subcellular fractionation of liver homogenates from cholesterol-sphingomyelin treated rats followed by lipid and enzymatic analyses of the fractions revealed that most of the accumulating lipid was present in very low density lysosomes floating in the post-microsomal supernatant fraction. The low density lysosomes exhibited good latency and had a very much lower relative activity of sphingomyelinase compared with values for N acetyl-beta-glucosaminidase. Multiple injections of sphingomyelin-cholesterol liposomes resulted in splenomegaly. The spleen homogenates of the treated rats showed a many-fold increase in the concentration of sphingomyelin, of non esterified cholesterol and in the activity of N-acetyl-beta-glucosaminidase over corresponding values for rats injected with saline. Electron microscopy of liver and spleen sections from treated rats revealed distinctive polymorphic intracellular inclusions bound by a membrane and containing numerous osmiophilic bodies. Structures identical to the storage inclusions seen in fixed liver sections from treated rats were also seen by electron microscopy in the post microsomal fraction of these livers. The results suggest that sphingomyelin and non-esterified cholesterol accumulate in lysosomes when they occur in cells in excess of that structurally associated with cellular membranes. PMID- 2992981 TI - Independent lateral diffusion of cytochrome bc1 complex and cytochrome oxidase in the mitochondrial inner membrane. AB - Distinct fluorophores have been conjugated to antibodies for cytochrome bc1 complex and cytochrome oxidase, two integral electron transferring proteins in the mitochondrial inner membrane. Addition of these fluorescent antibodies to preparations of mitochondrial inner membranes followed by appropriate secondary antibodies causes distinct and independent aggregation of the two cytochrome proteins. These results reveal that both cytochrome bc1 complex and cytochrome oxidase diffuse laterally in the membrane plane independent of one another consistent with the random collision model for electron transport in the mitochondrial inner membrane. PMID- 2992982 TI - Morphological integrity of single adult cardiac myocytes isolated by collagenase treatment: immunolocalization of tubulin, microtubule-associated proteins 1 and 2, plectin, vimentin, and vinculin. AB - Single cardiac myocytes were isolated from hearts of 9 to 12-week-old rats by means of collagenase (100 U/ml). After assessment of their functional integrity they were processed for immunofluorescence microscopy of the cytoskeletal proteins tubulin, microtubule-associated proteins 1 and 2 (MAP-1 and MAP-2), plectin, vimentin, and vinculin. Antibodies to tubulin decorated a delicate filamentous network that apparently was unrelated to any sarcomeric organization. The distribution of MAP-1 and MAP-2 was strikingly different from that of tubulin, as both antigens were confined to Z-line structures. These structures were also prominently stained by affinity-purified antibodies to plectin and a monoclonal antibody to vimentin. Co-distribution of plectin and vimentin was also observed at the former intercalated disk region of the heart cell. Anti-vinculin antibodies decorated an intricate meshwork consisting of delicate filaments with predominantly irregular orientation and occasional assembly into whorls. These immunolocalization data indicate that the cell shape and cytoskeletal architecture characteristic of cardiac myocytes in tissues is maintained in single isolated cells. Furthermore, intermediate filaments rather than microtubules seem to be instrumental in the preservation of cell morphology. PMID- 2992985 TI - Malignant fibrous histiocytoma. Report of a case and review of the literature. AB - Malignant Fibrous Histiocytoma is a sarcoma characterized by two different principal cell types, which has its origin in mesenchymal undifferentiated clones. So it must be included in the differential diagnosis of the very rare primary breast tumors with stromal origin. The review of available literature allows us to define the histopathological and biological peculiarities of this kind of tumor and it confirms the extreme malignity and rarity of the case we observed. PMID- 2992983 TI - Localization of the Na+/K+-ATPase and of an amiloride sensitive Na+ uptake on thyroid epithelial cells. AB - The Na+/K+-ATPase was localized using purified specific antibodies, on the basolateral membranes of rat thyroid epithelial cells and of cultured porcine thyroid cells, by immunofluorescence and immunoelectron microscopy. No staining was observed on the apical membranes. When cultured cells formed monolayers, with their apical pole in contact with the culture medium, 22Na+ uptake was inhibited by amiloride. Inhibition was dependent upon extracellular Na+ concentration, half maximal inhibition was obtained with 0.7 microM amiloride in the presence of 5 mM Na+. Ouabain was ineffective on Na+ uptake into intact monolayers. A brief treatment of the monolayers with ethyleneglycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) opened the tight junctions and allowed the access of ouabain to the basal pole of the cells. In this condition ouabain increased Na+ uptake. When cells were reorganized into follicle-like structures, with their basal pole in contact with the culture medium, Na+ uptake was not modified by amiloride but was increased by ouabain. We conclude that in thyroid cells, the Na+/K+-ATPase is present on the basolateral domain of the plasma membrane whereas an amiloride sensitive sodium uptake occurs at the apical surface. PMID- 2992986 TI - Breast carcinoma metastasized in ovarian thecoma (a case report). AB - We describe a patient with breast cancer metastatic to ovarian thecoma. This finding, to the best of our knowledge, has yet to be reported in literature regardless of the fact that there are many gynecological neoplastic associations. Our hypothesis is that this unusual finding is not accidental, but rather benign lesions may be natural resting sites for metastatic disease. An autopsy series is required to elucidate this incidence. PMID- 2992984 TI - Morphological and functional polarity of an epithelial thyroid cell line. AB - The thyroid epithelial cell line FRT in monolayer culture appeared to be strongly polarized by morphological criteria. Cells were connected by tight junctions, exposed microvilli toward the culture medium and formed domes at confluency. FRT cells were infected with vesicular stomatitis virus (VSV) and Sindbis virus and the budding polarity was examined 8 and 16 h after infection, respectively. VSV budding occurred preferentially from the basolateral domain of plasma membrane, while Sindbis virus budding was mostly apical. The distribution of VSV and Sindbis virus glycoproteins, as determined by the immuno-gold technique, correlated well with the budding polarity. Polarized budding was not observed in isolated cells in suspension. PMID- 2992987 TI - Primary carcinoma of the Bartholin gland. AB - The authors report a case of primary carcinoma of the Bartholin gland treated with a radical vulvectomy and bilateral inguinal lymphadenectomy. PMID- 2992988 TI - Bilateral subdural haematoma demonstrated at scintigraphy of the brain. AB - Bilateral subdural haematoma is not a rare disorder, but only a few cases diagnosed at brain scintigraphy have been reported. Bilateral subdural haematoma was demonstrated at scintigraphy in a 66-year-old man. Subsequent computerized tomography scanning of the brain also demonstrated the lesion. After bilateral burr-hole aspiration, the patient made a good recovery. PMID- 2992989 TI - Emission tomography with 99mTc-pyrophosphate in the diagnosis of acute myocardial infarction. AB - Electrocardiograms (ECG) and enzyme criteria are usually used to confirm the diagnosis of acute myocardial infarction in the case of chest pain. However, ECG is not always diagnostic. Elevated enzyme values may be due to causes other than myocardial infarction. In uncertain cases, the ECG and enzyme criteria can be supplemented by emission tomography, performed with technetium pyrophosphate that will accumulate in the site of infarction. Twenty-nine patients with suspected acute myocardial infarction were studied with emission tomography. Of these 12 had acute transmural infarction. Both enzyme tests and ECG were diagnostic in only 7 of these 12 cases, 4 had positive enzyme tests but a nondiagnostic ECG and in one case neither enzymes nor ECG were diagnostic. In 11 patients the infarcted myocardial area was detected with emission tomography. Six patients had acute nontransmural infarction. Only 2 of these had positive emission tomography. The chest pain was not due to infarction in 11 patients. All these patients had negative emission tomography. The sensitivity of emission tomography was 92% and specificity 100% in transmural acute infarction. In nontransmural infarction the specificity was only 33%. Emission tomography is a valuable diagnostic tool. It may be the decisive method when ECG and enzymes are not diagnostic. Emission tomography also shows the localization and size of the infarcted area in the myocardium. PMID- 2992990 TI - Improvement in neuromuscular transmission in myasthenia gravis by 3,4 diaminopyridine. AB - 3,4-Diaminopyridine (3,4-DAP), a potent potentiator of action potential evoked release of acetylcholine from presynaptic terminals in the neuromuscular junction was given i.v. and p.o. to two patients with myasthenia gravis. Effects were monitored electrophysiologically by repetitive nerve stimulation and by standardized clinical testing. Administration of 8 mg and 9 mg 3,4-DAP i.v. produced a clear improvement in the neuromuscular transmission after approximately 20 min. When 3,4-DAP was given p.o. 24 mg was shown to be effective. At a dosage of 18-24 mg p.o. 3,4-DAP significantly potentiated the effect of the cholinesterase inhibitor pyridostigmine at an optimal dose. The maximal effect of 3,4-DAP p.o. was obtained after 2.5-3 h. No significant CNS side-effects were found which is in contrast to those reported for 4 aminopyridine. The results suggest that 3,4-DAP may be useful as an addition to the conventional treatment with cholinesterase inhibitors when immunosuppressive treatment is considered contraindicated or when it has not yet reached its full effect. PMID- 2992991 TI - Hemichorea associated with varicella-zoster reinfection and endocarditis. A case report. AB - A 20-year-old woman developed transient right-sided hemichoreatic movements after household exposure to varicella-zoster. Some days before the appearance of involuntary movements a vesicular rash had occurred. About 6 months later an elevated IgG serum titer against varicella virus was found and two-dimensional echocardiography showed signs of an endocarditis. During the following 2 months the IgG value returned to within the normal range and the choreatic movements disappeared almost totally. The possibility is discussed that endocarditis had been caused and maintained by serum antibodies to varicella-zoster virus which cross-reacted with valvular tissue. Embolization to the region of the left striatum and/or postinfectious encephalitis in this region are assumed to be the most plausible causes of the transient hemichorea. PMID- 2992994 TI - Evidence that cyclic guanosine monophosphate (cGMP) mediates endothelium dependent relaxation. AB - The mechanism of action of endothelium-derived relaxant factor (EDRF) was studied using aortic strip preparations of the rabbit and a bioassay system of a rabbit coronary artery perfused in series with an intact aorta. Methylene blue (an inhibitor of guanylate cyclase) inhibited, and 2-O-propoxyphenyl-8-azapurine-6 one (MB22948, an inhibitor of cGMP phosphodiesterase) potentiated the vascular effects of EDRF whether these were due to its basal or to stimulated release. Infusion of these agents at different sites in the bioassay indicated that they act pharmacologically at the smooth muscle level and not on release of EDRF or by chemical interaction with EDRF. The data are consistent with the hypothesis that EDRF-induced relaxation is mediated by elevation of smooth muscle cGMP levels. PMID- 2992992 TI - Primary signet-ring cell carcinoma of the urinary bladder (linitis plastica). AB - In this paper we describe the 16th case in the English medical literature of signet-ring cell carcinoma of the bladder, which occurred in a 65-year-old man who developed widespread abdominal metastases. This case illustrates the characteristic diffuse infiltrative pattern of this neoplasm. In addition to light microscopy, ultrastructural studies were performed. PMID- 2992993 TI - Effects of (-)-N6-(R-phenylisopropyl)-adenosine (PIA) and caffeine on nociception and morphine-induced analgesia, tolerance and dependence in mice. AB - (-)-N6-(R-phenylisopropyl)-adenosine (PIA) was shown to possess analgesic activity in both the tail flick and acetic acid writhing assays. The analgesic actions of PIA were antagonized by caffeine in a dose-dependent manner. An apparent pA2 analysis in vivo suggested that the antagonism by caffeine was not competitive. Subanalgesic doses of PIA potentiated morphine-induced analgesia, tolerance and dependence. Caffeine antagonized these effects of morphine. PIA attenuated while caffeine exacerbated opiate withdrawal. While a low dose of caffeine antagonized PIA effects on withdrawal, a low dose of PIA did not antagonize the effects of caffeine. These results indicate that PIA can facilitate, and caffeine can antagonize the actions of morphine and that caffeine may be exerting some of its actions independent of adenosine receptor antagonism. PMID- 2992995 TI - Neuronal mechanisms involved in drug-induced jumping behavior in mice. AB - Previously we have found that lithium chloride (250 mg/kg i.p.) plus haloperidol (4 mg/kg i.p.), or apomorphine (0.25 mg/kg i.p.) plus thyrotropin releasing hormone (TRH, 20 mg/kg i.p.), elicited a jumping behavior which involves dopaminergic and cholinergic inhibition, and noradrenergic activation. Pretreatment with antiserotonergic agents such as methysergide (5 and 10 mg/kg i.p.) and cyproheptadine (5 mg/kg i.p.) enhanced the jumping behavior induced by these drugs. 5-Methoxy-N,N-dimethyltryptamine (5-MDMT, 5 mg/kg i.p.), a serotonergic receptor agonist, inhibited the jumping behavior. Muscimol, a GABA receptor agonist, at 2 mg/kg, potentiated jumping. GABA receptor antagonists such as bicuculline (4 mg/kg i.p.) and picrotoxin (0.2 and 1 mg/kg i.p.) depressed jumping. An antihistamine agent, diphenhydramine (5 mg/kg i.p.), also potentiated the jumping behavior. Furthermore methysergide (10 mg/kg i.p.), but not muscimol (2 mg/kg i.p.) elicited jumping behavior when combined with clonidine (0.5 mg/kg i.p.), TRH (20 mg/kg i.p.), haloperidol (4 mg/kg i.p.) or atropine (5 mg/kg i.p.). These results suggest that in addition to dopaminergic, cholinergic and noradrenergic mechanisms, serotonergic inhibition may be directly contributing to the initiation of jumping behavior, whereas GABAergic activation appears to be a modulating factor in this behavior. PMID- 2992996 TI - Effects of pGlu-His-Pro-amphetamine (TRH-amphetamine) on the isolated duodenum of the guinea-pig: antagonistic effect of amphetamine on TRH response. AB - pGlu-His-Pro-dexamphetamine (TRH-A) produced a contraction through the release of acetylcholine from postganglionic cholinergic neurons in the duodenum of the guinea-pig in the same manner as TRH. However, the affinity of TRH-A (pD2, 4.70) toward isolated duodenum was one thousandth that of TRH (pD2, 7.74). The effects of TRH-A (10(-4)M) were abolished by 10(-7) M TRH, but only partially (about 50%) inhibited by 10(-4) M d-amphetamine. D-Amphetamine showed no stimulatory effect on the myenteric nerves. However, the duodenal response to TRH (10(-6) M) was dose dependently inhibited by d-amphetamine (10(-6), 10(-5), 10(-4) M) while the phasic and tonic contractions caused by high K+ (40 mM) or the contractile responses to acetylcholine (10(-7) M) were not blocked by d-amphetamine. These results indicate that d-amphetamine may act as an antagonist to TRH without influencing the movement of calcium ions in smooth muscle or muscarinic receptors and that contractile responses to TRH-A are mediated through TRH receptors in the myenteric cholinergic nerves. PMID- 2992997 TI - Increased myocardial alpha 1 adrenoceptor density in rats chronically treated with propranolol. AB - Chronic treatment with propranolol for 6 weeks resulted in a 16.6% increase, over the level in control rats, in myocardial alpha 1-adrenoceptor density as measured by [3H]prazosin binding. The apparent affinity remained unaffected. No changes were observed in beta-adrenoceptor density and apparent affinity as measured by [3H]dihydroalprenolol binding. It is concluded that myocardial alpha 1 adrenoceptors might have a compensatory role under conditions where beta adrenergic effects are attenuated. PMID- 2992998 TI - PK 11195, an antagonist of peripheral benzodiazepine receptors, reduces ventricular arrhythmias during myocardial ischemia and reperfusion in the dog. AB - PK 11195, an antagonist of peripheral type benzodiazepine receptors, in doses from 5 to 25 mg/kg i.d. protected in a dose-dependent manner dogs against both early and delayed ventricular arrhythmias induced by 20 min ischemia and against ventricular fibrillation following reperfusion. Thus, peripheral-type benzodiazepine receptors might represent a novel target in the treatment of angina and cardiac ischemia. PMID- 2993000 TI - Increase in nigral type II benzodiazepine recognition sites following striatonigral denervation. AB - In the rat substantia nigra Type II benzodiazepine recognition sites (measured as the portion of [3H]flunitrazepam binding which remain after the addition of 2 X 10(-7) M Cl 218872) represent 50% of the total benzodiazepine recognition sites. The density of Type II sites was increased by 35% following the degeneration of the striatonigral afferents induced by the intrastriatal injection of kainic acid. On the other hand the same lesion failed to change the density of the remaining nigral Type I sites. The results indicate that denervation induces supersensitivity to nigral Type II benzodiazepine recognition sites. PMID- 2992999 TI - Multiple [3H]imipramine binding sites in brains of male and female Fawn-Hooded and Long-Evans rats. AB - Comparisons of high- and low-affinity [3H]imipramine binding to whole brain homogenates from adult male and female rats of the Fawn-Hooded and Long-Evans strains were performed. Most strikingly, no significant differences were observed between the two strains in any of the binding parameters, indicating that brain [3H]imipramine binding sites, which may be related to the serotonergic uptake process, appear normal in a strain of rats with serotonin platelet storage pool disease. However, a significant sex difference in high- but not low-affinity whole brain [3H]imipramine Bmax values was observed, with females of both strains having higher densities than males. The observed sex difference in densities of high-affinity [3H]imipramine binding sites necessitates further research into possible sex hormone interactions with this binding site and serotonergic transmitter systems. PMID- 2993001 TI - Inhibition of H+K+ATPase by SCH 28080 and SCH 32651. AB - The novel antiulcer agents, SCH 28080 and SCH 32651 were examined for their ability to inhibit the H+K+ ATPase enzyme activity in a preparation of microsomal membranes from rabbit fundic mucosa. SCH 28080 inhibited the isolated enzyme activity with a potency similar to omeprazole, IC50s of 2.5 and 4.0 microM respectively. SCH 32651 was less potent exhibiting an IC50 of 200.0 microM. Both compounds may therefore exert their antisecretory activity via a direct inhibition of the parietal cell H+K+ ATPase. PMID- 2993002 TI - Elevated arterial cyclic AMP levels during the development of spontaneous hypertension in rats. AB - Vascular cyclic AMP alterations were studied during the initiation of vascular hypertrophy and hyperplasia in spontaneously hypertensive rats (SHR). The onset of hypertension at 6 weeks of age coexisted with a three-fold elevation in the aortic content and concentration of cyclic AMP, whereas aortic DNA and protein contents were identical to those of WKY controls. A similar cyclic AMP elevation was present in 12-week-old SHR when vascular hypertrophy and hyperplasia were already established. These experiments suggest the participation of cyclic AMP in the process of hypertensive vascular growth. PMID- 2993003 TI - Human embryonal carcinoma cells express low levels of functional receptor for epidermal growth factor. AB - Using immunological probes, we have detected expression of the receptor for epidermal growth factor (EGF) at the cell surface of the clonally-derived human embryonal carcinoma (EC) cell lines, 2102Ep cl. 4D3 and NTERA-2 cl. D1. By this method, the level of receptor expression by these cells is estimated to be 3- to 5-fold less than for the human diploid fibroblast cell line, WI38, and our results indicate that it is the EC stem cells that display this receptor and not a subpopulation of differentiated cells. The human EC cell receptor binds ligand and catalyses autophosphorylation at tyrosine in a normal fashion. Treatment of NTERA-2 cl. D1 cells with retinoic acid (RA) for 7 days to induce differentiation results in decreased levels of receptor expression, and a subpopulation of differentiated cells possessing a markedly higher level of the EGF receptor was not detected among the cultures exposed to RA for longer periods. PMID- 2993004 TI - Biosynthesis of EGF receptor, transferrin receptor and colligin by cultured human keratinocytes and the effect of retinoic acid. AB - The biosynthesis of EGF and transferrin receptor by human keratinocytes in culture has been followed using specific monoclonal antibodies. In addition, keratinocytes are shown to synthesise a Mr 47 000 protein that binds to gelatin Sepharose. Peptide mapping confirms the identity of this protein with colligin, a newly described cell surface-associated glycoprotein that also binds to native collagens (Kurkinen et al., J biol chem 259 (1984) 5915) [9]. Vitamin A and its analogues have profound effects on the differentiation, morphology and motility of human keratinocytes in culture. We show here that retinoic acid (RA) has no effect on the growth rate of the cells or the synthesis of EGF receptor and colligin, but stimulates the synthesis of transferrin receptor. PMID- 2993006 TI - Induction of post-aggregative differentiation in Dictyostelium discoideum by cAMP. Evidence of involvement of the cell surface cAMP receptor. AB - Exogenous cAMP is known to induce post-aggregative differentiation in Dictyostelium discoideum under conditions that normal development is blocked. We have analysed the cyclic nucleotide specificity, the effect of modulation of the cAMP signal and the dose-response relationship of the induction of two independent markers of post-aggregative differentiation, i.e., a prespore cell specific antigen detected by a monoclonal antibody, and the activity of glycogen phosphorylase. Our results confirm that high concentrations of cAMP (10(-6)-10( 3)M) are required for the induction of these markers. The cells are shown not to adapt to the cAMP signal. The cyclic nucleotide specificity of induction agrees with the specificity of the cell surface cAMP receptor, but is very dissimilar to the specificity of the intracellular cAMP-dependent protein kinase. It is thus unlikely that cAMP leaks into the cell and activates the cAMP-dependent protein kinase directly. Instead, the induction of post-aggregative differentiation by cAMP seems to be mediated by cell surface cAMP receptors. PMID- 2993005 TI - Mitogenic activity of glia maturation factor. Interaction with insulin and insulin-like growth factor-II. AB - The mitogenic activity of glia maturation factor (GMF) was tested on sparse cultured cells. GMF stimulates the growth rate of normal astroblasts and fibroblasts grown in the presence of fetal calf serum (FCS), and raises the saturation density of the cells over what is imposed by the corresponding serum concentrations. GMF has no mitogenic effect in the complete absence of serum. The mitogenicity of GMF is also demonstrable in defined media where certain serum components are present. In particular, GMF in combination with the defined medium N2 partially mimics the proliferative effect of serum alone. Insulin, an ingredient of N2, can substitute for the complete N2 formula. Insulin-like growth factor-II (IGF-II), in turn, can substitute for insulin. The interaction of GMF with insulin or IGF-II can be demonstrated in a sequential manner, suggesting that GMF is a competence factor. Since insulin is required at a concentration well above the physiologic serum level, and must be used at a dose 1000 times higher than IGF-II, we suspected that insulin acts on IGF-II receptors. This was substantiated by the demonstration of IGF-II receptors and the absence of detectable insulin receptors on the astroblasts. The combined effect of IGF-II and GMF mimics the combined effect of 10% FCS and GMF, in both growth rate and saturation density. PMID- 2993007 TI - Efficient introduction of contents of liposomes into cells using HVJ (Sendai virus). AB - The conditions for efficient introduction of the contents of liposomes into cells were examined using fragment A of diphtheria toxin (DA) as a marker; one molecule of DA can kill a cell when introduced into the cytoplasm. Liposomes containing DA (DA liposomes) were toxic to cells treated with HVJ (Sendai virus) at 4 degrees C just before exposure to DA liposomes at 37 degrees C, but were not toxic to untreated cells. This toxicity was temperature-dependent. DA outside of liposomes was not toxic to HVJ-treated cells. Results also showed that liposomes could fuse with HVJ at 37 degrees but not at 4 degrees C and that liposomes preincubated with HVJ at 37 degrees C could associate with cells. DA liposomes preincubated with HVJ at 37 degrees C were highly toxic to cells. This toxicity was dependent on the duration of preincubation with HVJ and the dose of HVJ. When plasmid DNA coded herpes simplex virus thymidine kinase was trapped in liposomes and fused with Ltk- cells with HVJ, the thymidine kinase activity was expressed in about 10% of the cells. These data show that naked liposomes fuse efficiently with cells with HVJ and that the contents of the liposomes can be introduced into the cytoplasm 100-10 000 times more efficiently by treatment of the cells or liposomes with HVJ. PMID- 2993008 TI - Inhibitors of ADP-ribosyl transferase enhance the transformation of NIH3T3 cells following transfection with SV40 DNA. AB - The influence of inhibitors of the enzyme ADP-Ribosyl Transferase (ADPRT) upon the morphological transformation of NIH3T3 cells by calcium phosphate-SV40 DNA co precipitates was examined. Marked enhancement in the frequency of foci formation was noted when 3-methoxybenzamide was added to cells either during or after exposure to SV40 DNA, but there was no effect when cells were only pretreated with the inhibitor. The greatest enhancement was observed when cells were exposed to inhibitor both during and after transfection with DNA. The involvement of ADPRT activity in the events during transfection of DNA into cells is not understood. These results, however, highlight the importance of the key regulatory molecule poly (ADP-ribose) in the events which occur within the nucleus and which lead to transformation with SV40 DNA. PMID- 2993010 TI - Identification of low-Ca2+- and high-Ca2+-requiring neutral proteases in rat retina. AB - Two low-Ca2+-requiring proteases (calpain I) and one high-Ca2+-requiring protease (calpain II) have been separated from the cytosol of rat retina by DEAE-cellulose column chromatography. Calpain I was half-maximally activated at 3 microM free Ca2+ and fully activated at 10 microM free Ca2+. Half-maximal activation of calpain II was at 0.4 mM free Ca2+ while full activation was observed at 0.8 mM free Ca2+. Calpain activity has also been demonstrated in sealed rod outer segments. The soluble fraction of the rod outer segments contained 89% of the enzyme activity. The possible role of calpain in retina is discussed. PMID- 2993009 TI - Kallikrein and kininases in ocular tissues. AB - The retina, choroid, ciliary body, iris and aqueous humor of swine eyes contain enzymes capable of producing (kallikrein) and inactivating (kininase I and II) kinins. The activity of the enzymes varies in different eye structures. Higher activities of kallikrein and kininase II were found in highly vascularized tissues such as retina, choroid and ciliary body. The highest activity of kininase I (carboxypeptidase N) was found in the aqueous humor. The presence of these enzymes in the eye structures suggests a possible role for them in local metabolism of vasoactive peptides. PMID- 2993011 TI - The effect of delta 1tetrahydrocannabinol on luteinizing hormone release in castrated and hypothalamic deafferented male rats. AB - delta 1Tetrahydrocannabinol (THC) acutely suppresses tonic serum luteinizing hormone (LH) and prolactin levels in adult male rats. The exact site of its action has not been identified. We have performed complete hypothalamic deafferentation (CHD), which disrupts the medial basal hypothalamus (MBH) from the rest of the CNS, but did not abolish the ability of THC to suppress hypothalamic-pituitary responses in gonadectomized male rats. This was shown by the equal reduction in serum levels of LH and prolactin in non-deafferented (ND) and CHD animals. These results indicate that THC is able to act inside the MBH and that the MBH-pituitary axis remains responsive to its inhibitory effect despite interruption of the neural connections between the MBH and extrahypothalamic areas. However, the corticotropin releasing factor neurons in the MBH appear functionally impaired as a result of the transection and become unresponsive to the normally produced THC stimulation. Different patterns of action seem to govern the various hypophyseal hormones controlled by the hypothalamus, suggesting that the release of LH releasing hormone and prolactin inhibiting factor might be maintained by the activity of neurons surviving inside the island. PMID- 2993012 TI - Involvement of beta-adrenoreceptors in the shift of ocular dominance after monocular deprivation. AB - In the visual cortex of young kittens, the extent of ocular dominance shift following brief monocular lid suture was less than expected in the hemisphere which had been locally perfused with beta-adrenergic receptor blockers at a moderate concentration. The present result was not due to the effect similar to binocular deprivation, since "local anesthetic effect" of beta-adrenergic blockers was unlikely to be involved in the present paradigm. Thus, the present results are consistent with the idea that beta-adrenergic receptors within the visual cortex are necessary, at least in part, for the regulation of visuocortical plasticity. PMID- 2993013 TI - The significance of nucleus raphe dorsalis and centralis for thermoafferent signal transmission to the preoptic area of the rat. AB - Neuronal spike-trains were recorded extracellularly within the preoptic area, which is generally considered the center of information processing for thermoregulation. The neuronal responses were tested by thermal stimulation of the scrotal and abdominal skin of the rat. After a neuron had been identified as warm responsive, electrical stimulation and/or lesion were applied in medial midbrain to test the influence on the preoptic neurons. Post-stimulus-histograms were determined by using 600 single shocks. Furthermore a small area of the medial midbrain was electrolytically lesioned and the thermal response of the neuron was tested again. All lesion, stimulation and recording sites were examined histologically. Electrical stimulation of either nucleus raphe dorsalis or centralis influenced the responsiveness of the recorded preoptic neurons. The post-stimulus-histograms predominantly reveal polysynaptic ascending pathways. Lesion of either nucleus raphe dorsalis or centralis abolished the responsiveness of the recorded preoptic neurons. Lesions lateral to these nuclei were ineffective. By this it is concluded that both nucleus raphe dorsalis and centralis are essentially involved in the thermal signal transmission to the preoptic area. PMID- 2993014 TI - Direct effects of the adrenergic neurotoxin DSP4 on central opiate receptors: implications for neuroendocrine studies. AB - We have shown that the noradrenergic neurotoxin DSP4 can acutely abolish naloxone induced secretion of LH in the rat. A part of this influence is clearly related to an unexpected interaction with hypothalamic opiate ( [3H]-naloxone) binding sites. Injection of DSP4 (50 mg/kg) can severely reduce opiate binding assayed in vitro. In addition, the drug is very potent in blocking opiate receptors in vitro, as determined in slice and homogenate assays. Consideration must now be given to the possibility that many previous studies on acute effects of DSP4 undoubtedly involved the opiatergic system in addition to noradrenergic terminals. Thus, DSP4 is not the drug of choice for experiments designed to probe catecholamine-opiate interactions in the control of LH release. PMID- 2993015 TI - Feature extraction and tonotopic organization in the avian auditory forebrain. AB - In a neurophysiological study within the auditory centers of the mediocaudal telencephalon of the starling, 601 neurons were tested for auditory responses. 369 of these units responded to pure tones, noise bands, amplitude modulations (AM), or species-specific sounds. Of all the auditory neurons, 16.8% did not respond to pure tones but only to more complex stimuli (tone-unresponsive-, TU units). The remaining auditory units were classified as tone-responsive (TR units). In 44.3% of TR-units (i.e. 36.9% of all auditory units) differing responses to tones versus more complex stimuli were observed. Responses as they occur in TU-units and in the differing responses of TR-units can be explained by neuronal extraction of features in the time (108 out of 198 neurons) and in the spectral domain (82 out of 198 neurons). Responses to species-specific sounds usually can be explained in terms of extraction of these features. Among neurons sensitive to temporal features, exclusive responses to a narrow range of AM frequencies were observed. In those TU-units that represent spectral features some restrict their responses to noise bands with distinct bandwidths centered around a specific midfrequency. These units reject both wider and narrower noise bands. A tonotopic arrangement of auditory units is found in field L, the surrounding neostriatum (NCM), and the Hyperstriatum ventrale (HV). Isofrequency lines run as a continuum through NCM, field L, and the caudal part of HV. TU units are integrated into the tonotopic gradient according to the midfrequency of effective stimuli (e.g. noise bands or AM). The anatomical position of auditory units is correlated to their response properties. Within one isofrequency contour an increase in response selectivity is seen from field L to the postsynaptic areas in the NCM and the HV. The results are discussed in terms of possible mechanisms of feature extraction in the avian auditory system. PMID- 2993016 TI - Excitatory projections from hypothalamic and midbrain defense regions to nucleus paragigantocellularis lateralis in the rat. AB - Electrical stimulation of the hypothalamic and midbrain defense regions evoked convergent excitatory responses in neurons in nucleus paragigantocellularis lateralis (PGL) in rats anesthetized with urethane. Stimulation in PGL activated neurons in the periaqueductal gray matter (PAG) and in the hypothalamus antidromically. In the PAG the projecting neurons were localized in the defense region but in the hypothalamus they were more widely distributed lateral and dorsal to it, mainly in the lateral hypothalamic region and the zona incerta. We conclude that there are independent excitatory pathways from the midbrain and hypothalamic defense regions which relay on neurons in PGL. Stimulation in the midbrain defense region may activate the monosynaptic projection to PGL as well as polysynaptic pathways. In contrast, stimulation of the hypothalamic defense region probably excites mainly fibers of passage: the integrated pattern of autonomic changes produced by electrical stimulation may be due to activation of the efferent outflow from widely dispersed cell bodies whose axons run through the defense region. PMID- 2993018 TI - Stem cell concept and heterogeneity of malignant soft tissue tumor--a challenge to reconsider diagnostics and therapy? PMID- 2993017 TI - The intestinal profile of Na-K-ATPase in three rat models of acute renal failure. AB - The specific activity of mucosal Na-K-ATPase in segments of the small intestine and colon was examined after bilateral nephrectomies (BN), bilateral upper ureteral ligations (BUUL) and bilateral lower ureteral ligations (BLUL). Animals were studied 22-26 h after these respective operations. Bilateral nephrectomies and bilateral upper ureteral ligations resulted in an increase of the specific activity of the enzyme throughout the mucosa of the intestinal tract. Bilateral lower ureteral ligations induced no significant change in the specific activity of Na-K-ATPase in the intestinal mucosa. There were no differences in the degree of renal failure. Marked aldosteronemia was observed in the BN and the BUUL rats but not in the BLUL rats. These data suggest that the increase in the intestinal Na-K-ATPase activity in the BN and the BUUL rats may be related to the elevation of serum aldosterone as a regulator of the body potassium. PMID- 2993019 TI - [Interhemispheric pharmacological asymmetry]. AB - Based on primary change of the biochemistry of left hemisphere cortical neurons induced by aminazine and sodium bromide the conclusion is made about the presence of interhemispheric pharmacological asymmetry (IPA). The known data on the primary effect of alcohol on the right hemisphere are confirmed by the presence of the IPA. PMID- 2993021 TI - Transcriptional control of thyroglobulin gene expression by cyclic AMP. AB - Transcription of thyroglobulin (Tg) gene is under the positive control of thyrotropin (TSH). The mechanism of this control has been further investigated. Rats were treated with triiodothyronine (T3) to decrease their endogenous TSH production. Following the intravenous injection of bovine TSH, a 3-fold stimulation of Tg gene transcription could be detected in isolated nuclei as early as 1 h after treatment. The TSH effect was also observed in tissue fragments incubated in vitro under conditions where a concomitant stimulation of cAMP accumulation was detected. Forskolin, a universal activator of adenylate cyclase, was able to mimic TSH action on Tg gene transcription. We conclude that TSH controls transcription of Tg gene directly via its known interaction with receptors on thyrocytes and that cAMP is a physiological mediator of this effect. PMID- 2993020 TI - [Mechanism of action of fentolamin on the gastric secretion]. AB - Thirty-three experiments were made on mongrel dogs operated on according to Basov, with the use of phentolamine, pilocarpine, atropine, histamine and their combinations. It was established that administration of histamine gives rise to an increase in gastric secretion, provided the animals are pretreated with phentolamine or with combined pilocarpine and phentolamine. Histamine-induced gastric secretion considerably decreases, provided the animals are pretreated with atropine and ceases almost completely after administration of combined atropine and phentolamine. It is concluded that the stimulating effect of phentolamine on gastric secretion is caused by an increase in the sensitivity of H2-receptors and cholinoreceptors under blockade of the alpha-adrenoreactive systems. PMID- 2993022 TI - Vasoactive intestinal peptide receptor activity and specificity during enterocyte like differentiation and retrodifferentiation of the human colonic cancerous subclone HT29-18. AB - Commitment of HT29-18 cells to enterocyte-like differentiation by glucose removal is related to a decreased capacity to generate cAMP after treatment with vasoactive intestinal peptide (VIP), forskolin or sodium fluoride. In contrast, the potency of VIP (EC50 = 1.1 - 1.3 X 10(-10) M) and the pharmacological specificity of the VIP receptor (VIP greater than rh GRF 1-43 greater than PHI greater than secretin) are unchanged during differentiation and retrodifferentiation. These results indicate that disturbances in VIP receptor post-receptor activity, involving cell surface VIP receptors, membrane and intracellular transducers of hormonal information, occur during enterocyte-like differentiation of the HT29-18 subclone. PMID- 2993023 TI - Phorbol ester stimulates catecholamine synthesis in isolated bovine adrenal medullary cells. AB - In isolated bovine adrenal medullary cells, the phorbol ester 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C, stimulated [14C]catecholamine synthesis from [14C]tyrosine, but not from [14C]DOPA. This stimulatory effect of TPA on [14C]catecholamine synthesis was not dependent upon extracellular Ca2+, and TPA did not affect the uptake of 45Ca2+ or the release of catecholamine by the cells. TPA also did not affect the intracellular cyclic AMP (cAMP) level. 4 alpha-Phorbol 12, 13-didecanoate, which is not an activator of protein kinase C, did not stimulate the synthesis of [14C]catecholamine from [14C]tyrosine. The stimulatory effect of TPA on [14C]catecholamine synthesis was additive with that of carbamylcholine, but not with that of dibutyryl cAMP (DB cAMP). From these results, it was suggested that protein kinase C is involved in the regulation of tyrosine hydroxylase activity and that this regulatory mechanism might be similar to that involving cAMP. PMID- 2993024 TI - Dephosphorylation of the deinhibitor protein by the PCSH protein phosphatase. AB - The deinhibitor protein, responsible for the decreased sensitivity of the ATP,Mg dependent protein phosphatase to inhibitor-1 and the modulator protein, is inactivated by cyclic AMP-dependent protein kinase and reactivated by dephosphorylation. The specificity of this reaction was tested with the ATP,Mg dependent phosphatase in its activated or spontaneously active form, four different forms of polycation-stimulated phosphatases (PCSH, PCSM, PCSL and PCSC) and calcineurin. Only the high -Mr polycation-stimulated protein phosphatase (PCSH), but not its catalytic subunit (PCSC), shows a high degree of specificity for the deinhibitor protein. Deinhibitor phosphatase activity of PCSH is affected neither by polycations nor by Mn ions. PMID- 2993025 TI - Effect of phorbol ester and phospholipase C on LH-stimulated steroidogenesis in purified rat Leydig cells. AB - When the phorbol ester, 4 beta-phorbol-12-myristate-13-acetate (PMA) or bacterial phospholipase C (PL-C) is added to a preparation of purified adult rat Leydig cells, containing 2 mM CaCl2, a time- and dose-dependent decreases of LH stimulated testosterone production is observed. After a 3 h stimulation with oLH (100 ng/ml), PMA (100 ng/ml) and PL-C (1.6 U/ml) do not affect the cell viability or the hCG specific binding, while cAMP accumulation is significantly reduced; cAMP-stimulated steroidogenesis is diminished only in the presence of PL-C. These observations suggest that in vitro: (i) activated Ca2+- and phospholipid dependent protein kinase is implicated in the regulation of rat Leydig cell steroidogenesis by LH at a step before the adenylate cyclase; (ii) phospholipids play an important role in cAMP-stimulated testosterone synthesis. PMID- 2993026 TI - Regulation of a liver plasma membrane phosphoinositide phosphodiesterase by guanine nucleotides and calcium. AB - Rat liver plasma membranes are enriched in a Ca2+-dependent phosphodiesterase active on phosphatidylinositol 4,5-P2 and phosphatidylinositol 4-P, but not phosphatidylinositol. Inositol-P3 is the first product of the reaction, but is rapidly degraded. Micromolar concentrations of GTP and its nonhydrolyzable analogues stimulate the reaction, whereas GDP, GMP and other nucleoside triphosphates are inactive. GTP and its analogues decrease the requirement of the reaction for Ca2+ and also increase its activity at saturating Ca2+. These results support the hypothesis that guanine nucleotides and a guanine nucleotide binding regulatory protein are involved in coupling the receptors for Ca2+ mediated agonists to the breakdown of plasma membrane phosphatidylinositol 4,5 P2. PMID- 2993027 TI - Regulation of platelet cytosolic free calcium by cyclic nucleotides and protein kinase C. AB - PAF elicits a rapid, concentration-dependent elevation of platelet cytosolic free calcium ([Caf]), measured by quin2. Elevation of [Caf] is transient, and the rate of reversal increases with agonist concentration. Adenylate cyclase stimulants (PGI2, PGD2) and 8-bromo cAMP; a guanylate cyclase stimulant (sodium nitroprusside) and 8-bromo cGMP; and a protein kinase C stimulant (phorbol myristate acetate) block the elevation of [Caf] induced by PAF, and accelerate its reversal. These results suggest that cAMP, cGMP and 1,2-diacylglycerol (DAG) could act as second messengers to regulate [Caf] in platelets. As PAF is known to stimulate platelet phosphoinositide hydrolysis (ergo DAG formation) but fails to elevate platelet cAMP or cGMP, it is proposed that DAG, via activation of protein kinase C, may act as an endogenous modulator of platelet [Caf]: an action that contributes to the role of DAG as a bi-directional regulator of platelet reactivity. PMID- 2993028 TI - Differences in mechanisms of modulation between rat liver cholesterol 7 alpha hydroxylase and HMG-CoA reductase. AB - The effects of microsomal HMG-CoA reductase kinase, cytosolic phosphoprotein phosphatase and cytosolic, thiol-dependent cholesterol 7 alpha-hydroxylase stimulatory protein on purified cholesterol 7 alpha-hydroxylase and HMG-CoA reductase from rat liver were compared. Neither HMG-CoA reductase kinase nor phosphoprotein phosphatase had any significant effect on cholesterol 7 alpha hydroxylase activity. They inhibited and stimulated, respectively, the activity of HMG-CoA reductase. The purified cytosolic protein which stimulated cholesterol 7 alpha-hydroxylase threefold in the presence of glutathione had no effect on HMG CoA reductase. The results show that there are separate intracellular systems for modulation of cholesterol 7 alpha-hydroxylase and HMG-CoA reductase. PMID- 2993029 TI - Membranal tyrosine protein kinase activity (but not cAMP-dependent protein kinase activity) is associated with growth of rat mammary tumors. AB - DMBA induced rat mammary tumors were used to study the association of tyrosine protein kinase activity with tumor growth. Pharmacological manipulations of blood prolactin level, by perphenazine and bromocriptine, were used to stimulate or arrest tumor growth, respectively. During perphenazine treatment, a 2-3-fold increase in membranal tyrosine protein kinase activity, measured with angiotensin II as substrate, preceded the 3-4-fold increase in tumor area. At the same time the cAMP-dependent protein kinase activity, measured with kemptide as substrate, did not change. PMID- 2993030 TI - Polyphosphoinositide hydrolysis is associated with exocytosis in adrenal medullary cells. AB - [3H]inositol-labelled products are released from adrenal medullary cells during exocytotic secretion. In 'leaky' cells in which small molecules readily enter and leave the cytoplasm, addition of micromolar calcium ions in the presence of ATP stimulates exocytosis and causes the release of inositol polyphosphates. These data support the idea that hydrolysis of plasma membrane polyphosphoinositides may be an essential step in exocytotic secretion. PMID- 2993032 TI - Identification in several human myeloid leukemias or cell lines of a DNA rearrangement next to the c-mos 3'-end. AB - A characteristic DNA rearrangement, the loss of an EcoRI cleavage site next to the 3'-end of the human c-mos gene, has been found to be frequently present in DNA from transformed hematopoietic cells of the myeloid lineage but not in DNA from either normal or transformed cells of different tissue types. Three established cell lines, respectively a pro-monocytic line (CM-S) and two precursor granulocytic lines (My/K1 and My/K5), carry the same genome rearrangement, but not fibroblasts obtained from the marrow of the same patients. This DNA rearrangement is maintained in three different hybridomas derived by fusion of CM-S cells with normal human embryo hepatocytes. PMID- 2993031 TI - The dynamic nature of DNA-strand breaks present in differentiating muscle cells and quiescent lymphocytes. AB - Cellular differentiation in a number of eukaryotic systems is associated with changes in the number of DNA-strand breaks and involves the activity of adenosine diphosphoribosyl transferase (ADPRT). DNA-strand breaks are essential for activation of nuclear ADPRT, the activity of which is required for efficient religation of DNA-strand breaks. In this study we demonstrate the dynamic nature of DNA-strand breaks formed in the genome of differentiating avian skeletal muscle cells and quiescent human lymphocytes. Inhibition of ADPRT activity blocks DNA-strand ligation in both cell types and leads to the accumulation of a higher number of strand breaks. PMID- 2993033 TI - Is chemosensitivity testing for peri-operative treatment planning in gastro intestinal cancer by the human tumour colony assay worthwhile? AB - Based on the concept of a combined modality cancer treatment in surgical oncology, the use of the human tumour colony assay for routine chemosensitivity testing and prospective treatment planning was investigated in 204 surgical biopsies of primary human solid tumours. The majority of the tumours (135/204) were of gastro-intestinal (GI) origin. Sufficient growth for drug testing occurred in 29-67% of all tumours depending on the tumour type, with a mean of 36% in GI-carcinomas. Chemosensitivity testing in vitro against standard anti cancer agents correlated well with clinical experience, 5-FU and FUDR being the most active drugs (27% respectively 24% sensitive tumours in vitro) in GI carcinomas. Relatively good agreement of in vitro/in vivo correlations was seen with an overall of 25/32 correct predictions in GI and other tumours. Predictivity was particularly good for loco-regional chemotherapy. Nevertheless, the limited in vitro growth rate of gastro-intestinal tumour specimens and their chemoresistance restrict the use of this method-in particular with respect to individual treatment planning. PMID- 2993034 TI - Breast diseases in Jordanian females: a study of 1000 cases. AB - Pathological data on 1000 breast lesions obtained from Jordanian females were analyzed and compared to data available from other countries or ethnic groups. Ductal carcinoma was the most frequently encountered lesion followed by fibrocystic disease (mammary dysplasia), fibroadenoma and mastitis. The mean age of Jordanian females with ductal carcinoma was 44.5 years, and many patients presented with advanced stage of the disease as evidenced by the high frequency (74.6%) of nodal metastases in the patients who had axillary lymphadenectomy. The frequencies of medullary and mucinous (colloid) carcinoma were not greatly different from those in other countries, but lobular carcinoma had a substantially low rate of occurrence. Many patients with lactation-associated lesions such as mastitis, galactocele and lactating adenomas were noted, which is attributed to the high fertility rate in Jordan. PMID- 2993035 TI - Granular cell tumour of the breast. AB - Four cases of granular cell tumour of the breast reflecting the lifetime experience of one surgeon are presented. Recent advances in the pathology of this condition suggest that the customary term 'myoblastoma' is inappropriate, and that 'granular cell tumour' is preferable. The clinical and pathological features are reviewed and the differential diagnoses discussed, with emphasis on the distinction from carcinoma which may be mimicked both clinically and on frozen examination. The diagnosis may be made by pre-operative aspiration cytology, and a conservative surgical approach is recommended. PMID- 2993037 TI - Selective stabilization of retinotectal synapses by an activity-dependent mechanism. AB - How does each ingrowing retinal fiber select the right spot in the overall retinotopic projection? Chemospecific surface interactions appear to be sufficient only to organize a crude retinotopic map on the tectum during regeneration of the optic nerve of goldfish. Precise retinotopic ordering is achieved via an activity-dependent stabilization of appropriate synapses, based on the correlated activity of neighboring ganglion cells of the same receptive field type in the retina. Four treatments have been found to block the sharpening process: 1) blocking activity of the ganglion cells with intraocular tetrodotoxin (TTX); 2) rearing in total darkness; 3) correlated activation of all ganglion cells via stroboscopic illumination in a featureless environment; 4) block of retinotectal synaptic transmission with alpha-bungarotoxin. These experiments support a role for normal visually driven activity in sharpening the diffuse projection, and demonstrate that the correlated activity of the optic fibers interacts within the postsynaptic cells, probably through the summation of excitatory postsynaptic potentials. Intraocular TTX experiments suggest that a similar mechanism may drive both the formation of ocular dominance patches in fish tectum and kitten visual cortex and the segregation of different receptive field types in the lateral geniculate nucleus. Thus, it may be a general mechanism whereby the diffuse projections of early development are brought to a mature level of organization. PMID- 2993036 TI - Purified proteins acting on cultured chick embryo ciliary ganglion neurons. AB - Chick embryo ciliary ganglion neurons in dissociated monolayer culture have been used to examine molecular requirements for neuronal survival and neurite growth. These neurons will rapidly die in vitro unless supplied with an adequate level of ciliary neuronotrophic factor (CNTF), and even in the presence of CNTF they will not vigorously extend neurites on polyornithine substrata unless supplied with appropriate amounts of polyornithine-binding neurite-promoting factors (PNPFs). Recent work on the purification and partial characterization of embryonic chick eye CNTF and rat schwannoma PNPF is reviewed, and in vitro responses of ciliary ganglion neurons to other purified proteins such as laminin, fibronectin, insulin, and nerve growth factor are mentioned. PMID- 2993038 TI - Endogenous digitalis-like factors. Introductory remarks. PMID- 2993040 TI - Hyperprolactinemia in primary adrenocortical insufficiency. AB - A patient with primary adrenocortical deficiency presented with galactorrhea and hyperprolactinemia, in addition to the usual characteristic symptoms of adrenal insufficiency. With steroid replacement therapy, PRL levels returned to normal, with resumption of normal ovulatory cycles; and the patient became pregnant. Primary adrenocortical deficiency should be considered as one of the very rare causes in the differential diagnosis of hyperprolactinemia. PMID- 2993041 TI - [Participation of serotoninergic mechanisms in the regulation of visceral functions during conditioned reflex activity]. AB - In chronic experiments on dogs, a reverse correlation was revealed between the level of positive salivary conditioning and the amount of serotonin in the blood. Administration of 0.2 and 0.4 mg of serotonin into the anterior limbic cortex enhanced conditioning in animals with high initial amount of serotonin in the blood and decreased it in animals with low amount of serotonin in the blood. Subcutaneous administration of serotonin in doses 0.5, 1.0, 2.0 and 5.0 mg/kg augmented the small intestine electrical activity. Subcutaneous administration of the serotonin precursor 5-oxytryptophan (0.5 mg/kg) enhanced both the conditioning and the small intestine electrical activity. An increased level of serotonin in the blood was revealed in neuroses peculiar by a prolonged depression of conditioning. PMID- 2993039 TI - Relationships among endogenous digitalis-like factors in essential hypertension. AB - Elements of a hypothesis that relate endogenous digitalis-like factors to both natriuretic hormone and hypertension are briefly reviewed. The stimulus for secretion of these factors appears to involve a tendency toward a state of extracellular fluid volume expansion as a consequence of an inherited or an acquired defect in renal function. Several studies implicate the brain and, in particular, the hypothalamus in the control of the secretion. The digitalis-like factors are thought to act by partial inhibition of active sodium transport, thereby promoting increased intracellular levels of Na+ and Ca2+ in a variety of cell types. In the kidney, inhibition of sodium transport leads to a compensatory natriuresis to correct the tendency for volume overload. In smooth muscle, the inhibition of sodium transport will indirectly increase intracellular calcium levels. The increased availability of Ca2+ will elevate muscle tone and increase peripheral vascular resistance. Also presented are criteria that may be used to characterize digitalis-like activity in samples and extracts obtained from purification procedures. Finally, we review our measurements of the 6-h integrated plasma levels of digitalis-like factors and other hormones for normotensive subjects and patients with essential hypertension. The data indicate the presence of two classes of digitalis-like factors with potentially different roles in electrolyte metabolism and hypertension. PMID- 2993042 TI - [Changes in Na, K-ATPase activity following adrenoreceptor blockade]. AB - A regulatory factor released from the effector cell during blockade of postsynaptic alpha- and beta-adrenoreceptors with phentolamine and propranolol (5 X 10(7)-1 X 10(-6) M) and activating norepinephrine neuronal uptake and synthesis, was shown to activate, Na, K-ATPase of the rat isolated organs as well as the preparation of partly purified Na, K-ATPase extracted from the rat brain. Ouabain (1 X 10(-3) M) eliminated completely the activating effect of the regulatory factor on the ATPase. PMID- 2993043 TI - [Role of calcium ions and cyclic nucleotides in neurotrophic control of the membrane properties of muscle fibers in the frog]. AB - Subcutaneous injections of caffeine, calcium ionophore X537A or cAMP did not affect the changes of input resistance (R0) and time constant (T) of membrane caused by denervation of the frog m. sartorius but prevented the MP decrease and ACh sensitivity spread. Injections of cGMP did not affect the denervation changes of R0 and MP but increased the ACh sensitivity. The neurotrophic control of frog muscle membrane properties seems to depend on calcium and involve the cyclic nucleotides system. PMID- 2993044 TI - [Changes in the concentration of Na+ and K+ in the skeletal muscles of the rat during adaptation to cold]. AB - Intracellular Na+ concentration was found to increase in skeletal muscles of rats adapted to cold whereas K+ concentration decreased. The data obtained indicate a reduction of Na-pump capacity in skeletal muscles of rats adapting to cold. PMID- 2993045 TI - [Mechanisms of changes in cardiac output during stimulation of afferent fibers of a somatic nerve]. AB - In anesthetized cats, the technique of electromagnetic expenditure-metry aided to study changes of the cardiac output and venous inflow in pressor responses to stimulation of the tibial nerve's afferent fibers. The cardiac output was found to increase in 47% of observations which was accompanied by increased venous inflow in half of the cases and by decreased one in 1/3 of observations. Vagotomy as well as suppression of the efferent link of cardiac responses with atropin and obsidan induced shifts of the cardiac output in the somatic nerve stimulation. The optimum level of arterial pressure responses was revealed peculiar by positive shifts of the cardiac output in pressor responses to the above action. PMID- 2993046 TI - [Role of adrenergic mechanisms in manifesting the contractile effects of the smooth musculature of the ileum induced by serotonin]. AB - In acute experiments on cats, an isolated portion of the ileum was vascularly perfused with constant blood volume. Contractile activity of the ileum was estimated by isometric tension. The subthreshold dose of acetylcholine (6.10(-10) 6.10(-9) mol/ml/min) potentiated or inhibited the contractile responses of the ileum induced by i. a. administration of serotonin (2.5 X X 10(-8)-1.10(-7) mol). Blockade of beta-adrenergic receptors with propranolol (0.2 mg/kg) diminished, and blockade of alpha-adrenergic receptors with phentolamine (1 mg/kg) increased the serotonin-induced contractile responses. After beta-adrenergic blockade acetylcholine potentiated these responses in all experiments. After alpha adrenergic blockade acetylcholine mostly inhibited serotonin-induced contractile responses. The facilitating effect of acetylcholine on the serotonin-induced contractile responses of the ileum seems to stem from the changed balance between inhibiting adrenergic and activating cholinergic mechanisms, the latter being predominant. PMID- 2993048 TI - [Suction electrode for conducting morphophysiologic studies on fine and short nerve trunks in vitro]. PMID- 2993047 TI - [Thermoregulatory effect of beta-adrenoreceptor block in the cold-adapted rat at rest and during exertion]. AB - Inderal does not affect the thermoregulatory system in thermoneutral conditions. Blockade of beta-adrenoreceptors at 6 degrees C decreases the oxygen consumption in cold-acclimated rats and reduces it down to the level of control animals. Cold acclimated rats have the same level of work hyperthermia at different ambient temperatures (26 degrees C, 6 degrees C) and under inderal injection. Control animals can only the same in warm environment. PMID- 2993049 TI - What Delaware's physicians should know about HTLV-III antibody testing. PMID- 2993050 TI - Epidermal growth factor receptors in different skin tumors. AB - Specific binding of 125I-labeled epidermal growth factor (EGF) was measured in 62 skin tumors of different severity. Within a group of 28 benign tumors, 11 of 15 condylomata acuminata were receptor positive, whereas the investigated mesenchymal tumors and normal skin as a control were receptor negative. 6 of 18 basal cell epitheliomas bound EGF specifically. In the group of precancerous and malignant skin tumors, 7 of 8 squamous cell carcinomas had the highest number of EGF binding sites and a high affinity state, whereas 5 malignant melanomas were receptor negative. The clinical relevance of these findings is not yet clear due to the short follow-up of the patients. PMID- 2993051 TI - Acute polyhydramnios associated with fetal hepatoblastoma. AB - A 28-yr-old gravida II mother presented at 29 wk gestation with acute polyhydramnios. The most common causes for polyhydramnios were excluded and onset of preterm labor at 31 wk prevented further studies. Tocolysis failed and a severely asphyxiated boy was born. He had a very distended abdomen with a 7 cm hepatomegaly and was in shock. No other physical abnormalities were found. The child died 15 min after birth. At post mortem examination a fetal type hepatoblastoma was detected. Duodenal obstruction due to the liver tumor might have caused the polyhydramnios. PMID- 2993052 TI - Release of proteinases by cultures of human cell lines derived from squamous carcinomas of the tongue and larynx. AB - Seven human cell lines derived from squamous carcinomas of the tongue and larynx were examined for their ability to produce and secrete proteinases. All cell lines were able to release into the culture medium cysteine proteinase and plasminogen activator-like activities. All lines differed from each other in the amount of enzymes secreted, in the kinetics of the secretion, in the quality of the enzymes produced an in the intracellular pool of these activities. These features constitute potential criteria for classifying the biochemical behavior of the tumor cell lines and for explaining their different ability to invade soft tissues and bone. PMID- 2993053 TI - Inorganic pyrophosphate and inorganic pyrophosphatase in Escherichia coli. PMID- 2993054 TI - Incorporation and turnover of phospholipid precursors in normal and tumoral glial cells in culture. AB - The incorporation and turnover of phospholipid precursors in cultured normal and tumoral glial cells was investigated during the plateau phase of growth. Glycerol was incorporated similarly by all cell types, and was renewed with a half-life of 19-37 hr. Acetate had a much longer half-life in primary cultures (50-75 hr) than in proliferative tumor cells (20-40 hr). Phosphate had a more rapid turnover rate in primary cultures (25 hr) than in proliferative tumor cells (50 hr). For all precursors, inositol- and choline phosphoglycerides had a faster turnover rate than other phospholipids. PMID- 2993055 TI - Isolation and characterization of a cDNA and a pseudogene for mouse lactate dehydrogenase-A isozyme. AB - A mouse lactate dehydrogenase-A cDNA was isolated and it was shown to contain the 393bp of the protein-coding sequence and 488bp of the 3' untranslated region. The amino acid sequence deduced from its open reading frame provided independent evidence for the sequence of residues 201-331 of mouse LDH-A subunit (muscle). This cDNA clone was used as a probe to isolate a mouse genomic clone containing a truncated, processed LDH-A pseudogene. This pseudogene showed 81.6% homology at 713 positions compared with the LDH-A cDNA sequence. The divergence of this pseudogene was estimated to have occurred 39 million years ago. PMID- 2993057 TI - N-epsilon-acetylation of porcine mature erythrocytes ubiquitin. AB - Highly purified of porcine mature erythrocytes ubiquitin were obtained according to the experimental procedure reported by Jabusch and Deutsch (1983). N-epsilon acetylation in vitro of internal lysyl residues of ubiquitin by p-nitro-phenyl acetate at pH 8.0 was performed. The extent of acetylation of ubiquitin was determined: about 4-5 residues (4.5 residues) of N-epsilon-lysine groups of ubiquitin were acetylated. We have assigned by Edman degradation the sites of acetylation and the sites of remaining free internal N-epsilon-lysine residues in the sequence: fully acetylated: Lys-6, Lys-11 and Lys-33. Partially free N epsilon-lysine: Lys-27 and Lys-29 and probably Lys-48 and Lys-63. 50 cycles Edman degradation were performed on porcine ubiquitin and the first 45 N-terminal residues were identified. We have partially determined that the molecular conservation of 45 amino acid sequence of ubiquitin between cattle, man and swine since the 45 amino acid sequence out of 76 residues are identical. The amino acid composition between human and porcine ubiquitin are also identical. PMID- 2993056 TI - Uptake of sialo and asialo transferrins by isolated rat hepatocytes. Comparison of a heterologous and a homologous system. AB - Incubation of isolated rat hepatocytes with different human sialo transferrins shows that interaction with the specific transferrin receptor is insensitive to differences in the carbohydrate composition of the glycans. Asialo transferrins lead to an increased iron uptake, which is dependent on the amount of exposed galactose. This is explained by the presence of the asialo glycoprotein (AsGP) receptor. Experiments with selective saturation of the two receptor systems show that on incubation with human asialo transferrin (AsHTf) transferrin uptake proceeds increasingly via the AsGP receptor on raising the concentration. Homologous rat asialo transferrin (AsRTf) behaves similarly, but less pronounced. Iron is accumulated via both receptor systems in the heterologous system, but only via the transferrin receptors in the homologous system. The difference in interaction with the AsGP-receptor may be caused by the difference in galactose content of the two asialo transferrins. As an explanation for the differences in intracellular metabolism a hitherto unknown recognition system for species specificity is postulated which protects homologous AsRTf from degradation, but directs foreign AsTf to lysosomes. PMID- 2993059 TI - The applications of electron spin/paramagnetic resonance in biochemistry. Molecular Enzymology Group/Techniques Group/ Bioenergetics Group Joint Colloquium held in honour of Dr. Helmut Beinert. Leeds, 18-20 July 1984. PMID- 2993058 TI - Determination of inorganic pyrophosphate concentration in urine. AB - A simple method for the estimation of PPi in urine is described. The PPi and Pi may be determined simultaneously by this method. PMID- 2993061 TI - The electron paramagnetic resonance of metalloproteins. PMID- 2993060 TI - Electron-paramagnetic-resonance spectroscopy in biochemistry: past, present and future. PMID- 2993062 TI - Electron-paramagnetic-resonance studies using pre-steady-state kinetics and substitution with stable isotopes on the mechanism of action of molybdoenzymes. PMID- 2993063 TI - Electron-paramagnetic-resonance studies of Mn(II) complexes with enzymes and substrates. PMID- 2993065 TI - Electron-paramagnetic-resonance spectroscopy and related techniques in the study of nitrogenase. PMID- 2993064 TI - Electron-spin-resonance/electron-paramagnetic-resonance spectroscopy of iron sulphur enzymes. PMID- 2993066 TI - The use of electron-paramagnetic-resonance spectroscopy to establish the properties of nickel and the iron-sulphur cluster in hydrogenase from Chromatium vinosum. PMID- 2993067 TI - Electron-paramagnetic-resonance studies of photosynthesis. PMID- 2993068 TI - Spin-labels in biochemistry. PMID- 2993070 TI - Applications of spin-trapping to biological systems. PMID- 2993069 TI - Electron-spin-resonance studies on flavoenzymes. AB - Our current knowledge on the spin distribution of flavoenzyme semiquinones would suggest that it is very similar to that determined for model flavin systems. It remains for future work to establish whether this conclusion is valid for a wide range of flavoenzymes as well as for those in which the flavin has become covalently attached to the protein via the 8 alpha position. The improvement in e.s.r. instrumental capabilities, advances in ENDOR spectroscopy, and the application of other techniques such as electron spin echo spectroscopy should result in considerable new information regarding the structures and environment of flavoenzyme semiquinones. Further application of these physical approaches to flavoenzymes in which chemically modified flavins have been substituted for the 'normal' coenzyme (Massey & Hemmerich, 1980) may be particularly informative. PMID- 2993071 TI - The membrane-bound fumarate reductase of Escherichia coli: an electron paramagnetic-resonance study. PMID- 2993072 TI - Electron-paramagnetic-resonance studies on the spatial relationship of redox components in cytochrome oxidase. PMID- 2993073 TI - Electron-paramagnetic-resonance lineshapes of biological molecules: some effects of distributed parameters. AB - Great improvements in computer simulation of e.p.r. lines in several quite different cases has been achieved by replacing simple lineshape functions with distributions of more fundamental parameters. Analysis of the functional relationships between such parameters and field position has allowed explanation of unusual lineshapes in some cases and successfully predicted anomalous lineshapes in others. The descriptive parameters employed here may in turn be displaced by more fundamental parameters; in the future, more sophisticated lineshape analysis may lead to meaningful quantification of molecular conformational flexibility. PMID- 2993074 TI - Paramagnetic probes of multicomponent electron-transfer systems. AB - Electron-transfer systems associated with steroid hormone production in the adrenals and with photosynthetic NADP+ reduction in the chloroplast share some important features. Complexes between components in both systems affect the exposure of prosthetic groups to water-soluble paramagnetic groups. The sensitivity of the components to probes of different charges suggests that electrostatic interactions are important in complex formation. Haem plane orientation in cytochrome P-450 of the adrenal system is parallel to the membrane plane in both native and reconstituted systems. PMID- 2993075 TI - Electron-paramagnetic-resonance studies of structure and function of the two-haem enzymes Pseudomonas cytochrome c peroxidase and beef heart cytochrome c oxidase. AB - Beef heart cytochrome c oxidase contains two cytochromes, a and a3, and Pseudomonas aeruginosa cytochrome c peroxidase has one high- and one low potential c haem, cHP and cLP. The parallelism in co-ordination and spin states between cytochrome a and haem cHP on the one hand and between cytochrome a3 and haem cLP on the other is illustrated. The two latter haems become accessible to cyanide, when the former are reduced. Such reduction also leads to an activation of the enzymes. Mechanisms are presented in which ferryl forms of cytochromes a3 and haem cLP take part. The enzymes reach an oxidation state, formally the same as resting enzyme, but with different properties. PMID- 2993076 TI - Mitochondrial myopathies: deficiencies localized to complex I and complex III of the mitochondrial respiratory chain. PMID- 2993077 TI - Cytochrome c oxidase deficiency. PMID- 2993078 TI - Intramitochondrial loci for the hormonal regulation of hepatic gluconeogenesis. PMID- 2993079 TI - Inhibition of granulosa cell differentiation by dioctanoylglycerol--a novel activator of protein kinase C. AB - The actions of dioctanoylglycerol, a novel and specific activator of protein kinase C, on granulosa cell steroidogenesis and LH/hCG receptor induction were examined in vitro. Granulosa cells were cultured for 2 days in media containing 100 nM androstenedione. FSH treatment stimulated progesterone, 20 alpha-hydroxy pregn-4-en-3-one (20 alpha-OH-P) and estrogen production by 60- to 80-fold over basal levels. Treatment with dioctanoylglycerol increased progesterone production 3-fold over basal levels but failed to affect 20 alpha-OH-P or estrogen production. Concomitant treatment of cells with FSH and increasing amounts of dioctanoylglycerol produced dose-dependent inhibition of FSH-stimulated progesterone, 20 alpha-OH-P and estrogen production with IC50 values of 75, 40 and 50 microM, respectively. Dioctanoylglycerol also inhibited production of all three steroids induced by forskolin, an activator of adenylate cyclase, and dibutyryl cAMP. The effects of dioctanoylglycerol on cAMP responses to FSH were also examined. The increase in both intracellular and extracellular cAMP at 1 h following treatment with FSH was suppressed in the presence of dioctanoylglycerol as was the extracellular accumulation of cAMP at 48 h of culture. FSH induction of LH/hCG receptors, known to be a cAMP-mediated event, was also inhibited in the presence of dioctanoylglycerol. In contrast, a thio-derivative of dioctanoylglycerol (3-thio-1,2-dioctanoylglycerol), which does not activate protein kinase C, failed to inhibit both steroidogenesis and LH/hCG receptor induction in response to FSH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993080 TI - Characterization of gonadotropin-sensitive adenylate cyclase activity in human testis: uncoupling of the receptor-cyclase complex by specific hormonal antagonist. AB - Basal and gonadotropin stimulated adenylate cyclase activity was assessed in testicular tissues obtained from men (20-80 years). A disparity was observed in the gonadotropin responsiveness of the human testicular adenylate cyclase system to hFSH and hCG stimulation. Of the tissues analyzed, 61% were FSH responsive and 22% showed low response to hCG. Forskolin, a diterpene which activates adenylate cyclase by a receptor independent mechanism, stimulated adenylate cyclase activity in the gonadotropin unresponsive tissues. This suggests that the tissue unresponsiveness is due to an uncoupling of the catalytic subunit of the adenylate cyclase. Several functional properties of the FSH responsive human testicular adenylate cyclase were investigated. hFSH and oFSH stimulated the enzyme activity in a concentration dependent manner. However, the hormone (DG oFSH) in which 80% of the carbohydrate residues had been removed was inactive, despite its good binding ability to the FSH receptor. hFSH stimulated adenylate cyclase activity was inhibited by DG-oFSH but not by DG-hCG (deglycosylated hCG). The data demonstrates the existence of specific FSH and LH(hCG) receptors in human testicular membranes. The FSH receptors in some tissues are coupled to adenylate cyclase. The link between the FSH receptor and adenylate cyclase may be uncoupled in the presence of the deglycosylated form of oFSH resulting in a loss of hormone response. PMID- 2993082 TI - Transient biochemical compartmentalization of Purkinje cells during early cerebellar development. AB - It has recently been observed that during early cerebellar development--from embryonic Day 17 to postnatal Day 3 in the rat--only certain discrete clusters of Purkinje cells (PCs) are immunoreactive to cyclic GMP-dependent protein kinase (cGK). In contrast, at later stages and in the adult, all the PCs are immunoreactive. These results obtained with cGK suggest a transitory intrinsic heterogeneity in the immature cerebellar cortex. It seemed therefore interesting to investigate the distribution of other PC markers during early development in the rat and in other species. The results presented here were obtained with two other antibodies--against vitamin D-dependent calcium binding protein and against Purkinje cell specific glycoprotein--which, like cGK, label all adult PCs. Each antibody gave a different and reproducible mosaic of positive and negative clusters of PCs in the perinatal cerebellum, thus indicating a transient biochemical compartmentalization resulting from the differential expression of parts of the same genotype by clusters of PCs. This compartmentalization in concomitant with the ingrowing of the cerebellar afferents. Once synaptogenesis starts, the biochemical heterogeneity of PCs disappears. PMID- 2993081 TI - Morphological and functional characteristics of rat Leydig cells isolated on Percoll gradients: is Leydig cell heterogeneity in vitro an artifact? AB - Rat testicular intertubular cells have been isolated on Percoll density gradients. Detailed light and electron microscopic studies have determined the sedimentation positions for Leydig cells, macrophages, fibroblasts, endothelial cells, germ cells and residual bodies. Stereological techniques have been utilized to determine the number of cells in the region of the gradient where Leydig cells sediment. Morphologically intact Leydig cells were present in the more dense region of the gradient (1.0590-1.0900 g/ml), and they responded to hCG stimulation with an 11-fold increase in testosterone production and contained LH/hCG receptors. Leydig cells in the less dense region of the gradient (1.0440 1.0589 g/ml) secreted less testosterone and contained less hCG receptors than those obtained from denser regions. However, the morphological studies described herein provide evidence for the first time that the majority of these less functional 'Leydig cells' from the lighter region of the gradients do not contain a nucleus and represent pieces of Leydig cell cytoplasm with variable size, shape and complement of organelles. PMID- 2993083 TI - Insulin resistance of fat cells from spontaneously diabetic KK mice. Analysis of insulin-sensitive phosphodiesterase. AB - In an attempt to determine the mechanism of insulin resistance in the presence of obesity, we examined effects of insulin on insulin-sensitive phosphodiesterase (PDE) in spontaneously diabetic KK mice. Isolated fat cells prepared from epididymal adipose tissue were incubated, with or without insulin, for 10 min. In the case of subcellular fractionation, only membrane-bound PDE was activated by insulin, as was noted in the case of rat fat cells. The specific activity was decreased in KK mice compared with control C57BL/6 mice. The dose-response curve, expressed as a percent of the maximal insulin effect, shifted to the right and the increase of ED50 indicated a decreased insulin sensitivity in the KK mice. The maximal insulin effect did not change, either when expressed as a percent of the basal enzyme activity or when expressed on a per cell basis. Specific binding of [125I]-insulin in fat cells increased in KK mice and curvilinear Scatchard plots showed an increase of the high-affinity sites. These data indicate that impairment of PDE activation in fat cells of KK mice relates to postreceptor defects and the uncoupling may result in a decreased sensitivity. PMID- 2993084 TI - Plasma pancreatic polypeptide response to insulin-induced hypoglycemia as a marker for defective glucose counterregulation in insulin-dependent diabetes mellitus. AB - Defective glucose counterregulation occurs in some insulin-dependent diabetic subjects (IDDMs) as a result of a combined deficiency of glucagon (IRG) and epinephrine (EPI) secretion in response to insulin-induced hypoglycemia. To determine whether the deficient glucagon response, the deficient epinephrine response, or both are manifestations of autonomic dysfunction, we used the pancreatic polypeptide (PP) secretory response to insulin-induced hypoglycemia as a marker for autonomic neuropathy. Seven nondiabetic controls and 21 IDDMs were given insulin at 40 mU/kg/h after overnight euglycemia. Eight of the IDDMs had defective counterregulation (-CR), and 13 had adequate counterregulation (+CR) by our previously published criteria. Those with -CR had a blunted EPI (delta EPI = 102 +/- 16 pg/ml; mean +/- SEM) and PP (delta PP = 12 +/- 13 pg/ml) response as compared with controls (delta EPI = 310 +/- 49; delta PP = 498 +/- 43) and IDDMs with +CR (delta EPI = 291 +/- 32; delta PP = 521 +/- 86). In controls, IRG rose by 31 +/- 6 pg/ml; in IDDMs, IRG failed to rise significantly above baseline regardless of counterregulatory status. Although the PP and EPI responses correlated well (r = 0.626, P less than 0.001), the IRG response failed to correlate with either the EPI or the PP response. We conclude that the deficient epinephrine, but not glucagon, secretory response to hypoglycemia in diabetic subjects is a result of autonomic neuropathy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993085 TI - Virus-induced diabetes mellitus. No evidence for immune mechanisms in the destruction of beta-cells by the D-variant of encephalomyocarditis virus. AB - A possible contribution of the immune system to the pathogenesis of virus-induced diabetes mellitus was investigated using the D-variant of encephalomyocarditis (EMC-D) virus. Studies on the F1 and backcross progeny of susceptible and resistant strains of mice gave no suggestion of a linkage between susceptibility and the major histocompatibility locus. Immuno-suppression by antilymphocyte serum did not prevent the induction of EMC-D-induced diabetes. Athymic nude mice infected with EMC-D virus showed a nearly identical diabetogenic response as compared with heterozygous littermates. Passive transfer of lymphocytes from mice made diabetic with EMC-D virus into normal mice failed to produce diabetes. From these and other studies, we conclude that the development of EMC-D-induced diabetes is due to the direct destruction of beta-cells by the virus and that the contribution of the immune response to the pathogenesis of this disease is, at the most, minor. PMID- 2993086 TI - Maturation of cutaneous reflex responses recorded in the lower limb in man. AB - Cutaneous reflex responses from tibialis anterior muscle following electrical stimulation were recorded for 39 normal children between the ages of one and 16 years. In the first year of life stimulation produces a monophasic excitatory reflex response at short latency; in the second year it produces long- as well as short-latency increases in activity. Maturation of reflex function continues during the school years, with progressive emphasis on long- rather than short latency components. These results are discussed in relation to mechanisms underlying the acquisition of motor skills during childhood. PMID- 2993087 TI - Cutaneous reflex responses recorded in children with various neurological disorders. AB - Cutaneous reflex responses were recorded from tibialis anterior or first dorsal interosseous muscles of children with hemiplegia, spinal-cord compression, necrotizing sacroid granulomatosis, acute encephalomyelitis, myalgic encephalomyelitis, and a group of children attending the Learning Difficulties Clinic. Abnormalities of response are reported and are compared with the different reports in the literature of abnormal reflex EMG responses recorded by various methods. It is concluded that cutaneo-muscular reflex testing may have a part to play in the diagnosis of difficult paediatric problems. PMID- 2993088 TI - The influence of intra-uterine growth retardation on brainstem development of preterm infants. AB - This study examined brainstem function in 76 appropriate-for-gestational-age (AGA) and 25 small-for-gestational-age (SGA) infants born at less than 35 weeks gestation, using brainstem auditory evoked responses. During the preterm period the mean brainstem conduction time (BCT) of the 25 SGA infants was significantly shorter than that of AGA babies of the same gestation. The BCTs of the AGA infants decreased rapidly during postnatal development to term-equivalent age; those of the SGA infants did not change significantly. It is likely that prenatal factors are responsible for the alteration of early development in the neural function of non-asphyxiated SGA infants. This change in neural development may be important in determining later neurological performance. PMID- 2993089 TI - Achalasia secondary to nongastrointestinal malignancies. AB - Secondary or "pseudo" achalasia of the esophagus can mimic idiopathic achalasia radiographically and can be difficult to diagnose. Typically, it is due to invasive carcinoma involving the gastroesophageal junction, usually gastric adenocarcinoma. Occasionally, an achalasialike condition can be produced by tumors not involving the gastroesophageal junction. We report 2 cases, 1 of lung carcinoma and the other of hepatoma, in which the patients had radiographic and endoscopic changes compatible with achalasia. However, the onset of symptoms was abrupt and the patients were elderly; these are unusual features for primary achalasia. There have been several other reports of nongastrointestinal neoplasms producing a clinical and radiographic picture similar to achalasia. Although there are several theories as to the cause, our cases would support the concept that direct tumor involvement of the gastroesophageal junction is not necessary to produce significant esophageal dysmotility. PMID- 2993090 TI - Ca2+ channel blockers stimulate ileal and colonic water absorption. AB - The effects of calcium channel blockers on water transport in the rat ileum and distal colon were studied in vivo using the single-pass perfusion technique. Parenteral but not intraluminal verapamil, and parenteral nifedipine increased ileal water absorption, with effects lasting at least 60 min. In contrast, i.p. verapamil had no effect on rat distal colonic water absorption, whereas intraluminal verapamil significantly stimulated colonic water absorption. Similarly, perfusing the rat descending colon with low-Ca2+ Ringer's-HCO3 stimulated colonic water absorption. Verapamil was not antisecretory because the theophylline-induced decrease in ileal water transport was similar in control animals and in animals pretreated with i.p. verapamil. In addition, nifedipine stimulated active Na and Cl absorption in rabbit ileum. These studies demonstrate that the Ca2+ channel blockers verapamil and nifedipine stimulate basal absorption of water in rat ileum and distal colon in vivo, and stimulate active Na and Cl absorption in rabbit ileum in vitro. The verapamil stimulation of colonic water absorption from the luminal surface was duplicated by perfusion with a low-Ca2+ bathing solution. This suggests the presence of apical membrane Ca2+ channels in rat colon, which appear to be involved in regulation of basal water transport, and that these Ca2+ channels are in a partially open state under basal conditions. Because verapamil stimulates absorption systemically (ileum) as well as intraluminally (colon), Ca2+ channel blockers have properties that might be useful in treatment of diarrheal diseases. PMID- 2993091 TI - Malignant fibrous histiocytoma of the liver. AB - A 59-yr-old man was admitted to the hospital for evaluation of right upper quadrant pain, anorexia, weight loss, and low-grade fever of 2-mo duration. During his evaluation, an abnormal liver ultrasound and computed tomography scan demonstrated what proved to be an avascular hepatic lesion. At surgery, the diagnosis of malignant fibrous histiocytoma was established. We present herein our findings of what we believe to be the first reported study of malignant fibrous histiocytoma of the liver. PMID- 2993092 TI - Intrahepatic cholangiocarcinoma associated with hepatolithiasis. PMID- 2993093 TI - [Morphological prognosis factors in ovarian carcinoma]. AB - Ovarian tumours form a broad spectrum of histological types which reflect significant differences with regard to their biological behaviour and their responsiveness to radiological and cytostatical therapy. Precise subclassification is a precondition for selective therapy. A special subclass of tumours is represented by so-called borderline tumours which form a significant proportion of the overall group of epithelian ovarian neoplasms. Borderline tumours are histologically defined tumours which are neither fully benign nor overtly malignant. Though capable to metastasise they are of low malignant potential and favourable prognosis. In clinical statistics borderline tumours should be registered as a special entity. Within the ordinary group of malignant epithelial tumours histological grading has proven to be of prognostic significance. Amongst a great spectrum of useful cyto-histological criteria, nuclear polymorphism. The number of mitoses, and necrosis are the most important predictive factors. PMID- 2993094 TI - [Importance of clinically prognostic factors in the treatment of advanced ovarian carcinoma]. AB - 25 patients with stage III and IV ovarian carcinoma were treated with radical surgery and postoperative chemotherapy. Analysis of the disease-free intervals and the survival rates indicated significant differences related to the stage of the disease. Furthermore patients with minimal residual disease (tumor mass less than 2 cm in diameter) had a far better prognosis than patients with extensive residual disease (greater than 3 cm). No correlation could be demonstrated between histological grading, in vitro proliferation rates, hormone receptor status, type of postoperative chemotherapy and survival rates. PMID- 2993095 TI - [Intraductal breast carcinoma. Clinical, x-ray and histological findings and their therapeutic consequences]. AB - 116 cases of early ductal breast cancer diagnosed during 1971 to 1981 were analysed. In 65 cases no invasion was detectable. 37 cases showed an early stromal invasion and in 14 cases the invasion was questionable. A palpable mass was seen in 32% of the noninvasive and in 46% of the early invasive cases. The most important mammographic signs were grouped, suspicious microcalcifications (87%). Multicentricity occurred in 26% of the noninvasive and 43% of the early invasive cases. Two cases of the noninvasive and two of the early invasive group had axillary metastasis. Positive nodes were seldom in early ductal carcinoma (3%) compared to 42 invasive comedo-carcinomas (36% positive axillary nodes). In the follow-up five local recurrences were detected in the noninvasive group and one each in the other groups. In the noninvasive group 17% local recurrences occurred after breast conserving modalities compared to 4% in mastectomy patients. As no reliable data for the selection and the results of breast preserving modalities are available now, mastectomy and axillary dissection may be the safest therapy. Only in small intraductal breast cancer (under 25 mm) the breast may be conserved (wide excision, segmental resection, quadrantectomy), if complete excision is carefully controlled by mammography and histology and follow up is guaranteed. PMID- 2993096 TI - Hydroxylapatite augmentation of a maxillary defect. PMID- 2993098 TI - Calcium transport systems in cardiac sarcolemma and their regulation by the second messengers cyclic AMP and calcium-calmodulin. AB - The purpose of this survey is to describe the importance of cyclic AMP and Ca2+ calmodulin as mediators of the effects of beta-adrenergic agonists on cardiac sarcolemma. First, the basic characteristics of the three sarcolemmal Ca2+ transporting systems, the slow Ca2+ channel, the Ca2+-pumping ATPase and the Na+/Ca2+ antiporter, are described. These different pathways for in- and outflux of Ca2+ play a crucial role in the excitation-contraction coupling and relaxation of heart muscle. Catecholamines in the myocardium cause an increase in the rate and extent of tension development during systole, and in the rate of relaxation during diastole. These functional changes may largely be brought about by cyclic AMP-induced phosphorylation of membrane proteins that increases both the probability of opening the slow Ca2+ channels and the rate of Ca2+ pumping ATPase. It is generally believed that the effects on Ca2+ transport systems are due to direct actions of beta-adrenergic agonists leading to an increased cytosolic Ca2+ level during systole. Indirectly, an increase in systolic Ca2+ can amplify the primary effect of catecholamine on the Ca2+ pumping ATPase and probably also on the Na+/Ca2+ antiporter through Ca2+-calmodulin-dependent phosphorylation of membrane proteins. The intimate involvement of calmodulin in the operation of several sarcolemmal Ca2+-transporting systems is discussed in the light of the unknown mechanism of action of the so-called Ca2+ channel blockers, a class of drugs that have a very important potential to provide information on the fundamental reaction steps in excitation-contraction coupling. Some of these drugs are potent inhibitors of Ca2+-calmodulin-regulated enzymes. PMID- 2993099 TI - Mechanism of activation of Na, K-ATPase in nerve fibres during rhythmic excitation. AB - The mechanism of activation of Na, K-ATPase in nerve fibres during rhythmic excitation was studied. 3H-ouabain binding to the nerve was found to be dependent on the frequency of rhythmic excitation. During rhythmic excitation 3H-ouabain binding was increased in all nerves tested. The maximum of 3H-ouabain binding in squid and crab nerves was observed at 10 impulses/s, and in frog nerve at 100 impulses/s. The level of bound glycoside decreased during high-frequency excitation. Rhythmic excitation did not change Na, K-ATPase affinity to ouabain, but it appeared to increase the concentration of ouabain sensitive sites in the nerve membrane. The enhancement of 3H-ouabain binding to nerve during rhythmic excitation is interpreted as arising from transformation of "inactive" forms of the enzyme to "active" ones. PMID- 2993097 TI - Effects of caffeine on transport, metabolism and ultrastructure of isolated rat colon. AB - The effect of caffeine on the transport, metabolism and ultrastructure of the colon were determined. Segments of proximal colon were excised from the anesthetized rat and prepared for radioisotopic tracing of ion transport in the flux chambers or oxidative metabolism in an incubator. Other segments were fixed before or after caffeine administration for electron microscopy. The isolated rat colon actively transported both Na+ and Cl- in the absorptive direction, mucosa to serosa. Serosal addition of 10 mmol/l caffeine abolished the smaller Na+ transport but did not significantly affect the larger Cl- transport. The electrical potential difference and the short-circuit current rose accordingly. Although the oxidation of glucose was inhibited by 35%, caffeine had no significant effect on the oxidation of the fatty acid, butyric acid. Comparable metabolic responses were obtained using the isolated terminal ileum of the rat. Neither the height nor the density of the microvilli in the proximal colon were affected significantly by caffeine. It may be concluded that caffeine, unlike theophylline, effectively preserves the normal absorptive condition of the colon. Thus, caffeine may have actions other than inhibition of phosphodiesterase in the distal intestine. PMID- 2993100 TI - Length variation and heteroplasmy are frequent in mitochondrial DNA from parthenogenetic and bisexual lizards (genus Cnemidophorus). AB - Samples of mtDNA isolated from each of 92 lizards representing all color pattern classes of Cnemidophorus tesselatus and two populations of C. tigris marmoratus were digested with the restriction endonucleases MboI, TaqI, RsaI and MspI. The mtDNA fragment sizes were compared after radioactive labeling and gel electrophoresis. Three features were notable in the comparisons: there was little variation due to gain or loss of cleavage sites, two fragments varied noticeably in length among the samples, one by a variable amount up to a maximum difference of approximately 370 base pairs (bp) and the other by a discrete amount of 35 bp, these two fragments occasionally varied within, as well as between, samples. Two regions that corresponded in size to these variants were identified by restriction endonuclease cleavage mapping. One of these is adjacent to the D loop. Heteroplasmy, heretofore rarely observed, occurred frequently in these same two regions. Variability in the copy number of a tandemly repeated 64-bp sequence appears to be one component of the variation, but others (e.g., base substitutions or small additions/deletions) must also be involved. The frequent occurrence of these length variations suggests either that they can be generated rapidly or that they were inherited from a highly polymorphic ancestor. The former interpretation is favored. PMID- 2993101 TI - The locus encoding alpha A-crystallin is closely linked to H-2K on mouse chromosome 17. AB - We have used a complementary DNA (cDNA) for mouse alpha A-crystallin to probe genomic DNA for restriction fragment length polymorphisms which could be used to map the alpha A-crystallin gene locus (Acry-1) in the mouse genome. Ten of 12 restriction endonucleases produced fragment polymorphism among various inbred strains of mice. A comprehensive strain survey conducted with six endonucleases resulted in the discovery of six allelic forms of Acry-1. Linkage analysis was conducted on DNA from three sets of recombinant inbred strains of mice and demonstrated close linkage of Acry-1 with the major histocompatibility complex (H 2) on chromosome 17. Analysis of congenic and recombinant congenic strains of mice confirmed the linkage of Acry-1 and H-2 and located the alpha A gene to the region between glyoxylase (Glo-1) and H-2K. PMID- 2993102 TI - [Transposition immunity in bacteriophage Mu. The effect of a mutation at the kil gene on the establishment of immunity]. AB - Bacteriophage Mu is characterized by a phenomenon similar to the transposition immunity of TnA: the frequency of transposition of Mu or mini-Mu into plasmids containing certain phage sequences is reduced by two orders of magnitude. In order to lend transposition immunity to Mu, the recipient replicon must contain a sequence of phage DNA including a 5.1 kb early region from the c-end of Mu. The product of the kil (or cim) gene takes part in establishing the immunity. The transposition immunity of Mu is connected with the disturbance of cointegrate formation. PMID- 2993103 TI - Cloning and initial characterization of the metJ and metB genes from Salmonella typhimurium LT2. AB - The metJ and metB genes of Salmonella typhimurium have been cloned into Escherichia coli K-12 on a 19-kb EcoRI fragment in the plasmid vector pACYC184. The presence of a functional metB+ gene on this plasmid, designated pGS89, was demonstrated by its ability to complement a metB- E. coli mutant. The presence of a functional metJ+ gene on this plasmid was demonstrated by its ability to repress metC+ gene expression in a metJ- mutant transformed with this plasmid. The metJ gene product was identified in a minicell system as a polypeptide of Mr 12000. This polypeptide was not produced when the metJ gene was inactivated by insertion of a Tn5 element. Transformation of an E. coli metB- mutant with plasmid pGS89 (metB+, metJ+) results in transformants that grow slowly on glucose minimal medium or glucose-minimal medium supplemented with homocysteine. Methionine addition, however, restores normal growth. This phenotype requires the relA- mutation in the host strain and at least two other plasmid loci, one of which is the metJ+ gene. Transformation of an E. coli metJ- mutant with metJ- derivatives of plasmid pGS89 results in transformants that are unable to grow on either glucose-minimal medium or glucose-minimal medium supplemented with methionine. This phenotype requires the presence of a functional metB+ gene on the plasmid, and is unrelated to the status of the relA gene. PMID- 2993104 TI - Molecular cloning of the feline c-fes proto-oncogene and construction of a chimeric transforming gene. AB - The feline c-fes proto-oncogene, different parts of which were captured in feline leukemia virus (FeLV) to generate the transforming genes (v-fes) of the Gardner Arnstein (GA) strain of feline sarcoma virus (FeSV) and the Snyder-Theilen strain (ST) of FeSV, was cloned and its genetic organization determined. Southern blot analysis revealed that the c-fes genetic sequences were distributed discontinuously and colinearly with the v-fes transforming gene over a DNA region of around 12.0 kb. Using cloned c-fes sequences, complementation of GA-FeSV transforming activity was studied. Upon replacement of the 3' half of v-fesGA with homologous feline c-fes sequences and transfection of the chimeric gene, morphological transformation was observed. Immunoprecipitation analysis of these transformed cells revealed expression of high Mr fusion proteins. Phosphorylation of these proteins was observed in an in vitro protein kinase assay, and tyrosine residues appeared to be involved as acceptor amino acid. PMID- 2993105 TI - Sequence of the viral replicase gene from foot-and-mouth disease virus C1-Santa Pau (C-S8). AB - The nucleotide sequence of the region including the viral replicase gene, the carboxy terminus of protein P18, and the 3'-extracistronic region of foot-and mouth disease virus (FMDV) type C1-Santa Pau (C-S8) has been determined from previously cloned cDNA fragments [Villanueva et al., Gene 23 (1983) 185-194]. The comparison with the corresponding gene segments of FMDV of serotypes A or O shows base substitutions in 7.2-8.6% of residues in the replicase gene with no insertions or deletions. This is about fourfold lower variation than found for the region encoding capsid protein VP1 of the corresponding viruses. Intermediate variability (substitution at 16.1-23.6% positions) exists in the 3' extracistronic region, including point mutations, insertions and deletions. The predicted amino acid sequence of the replicase gene indicates that 75.5-82.6% of mutations are silent and that 93.4% of amino acids are conserved in the four FMDV replicases. The frequency of certain types of silent mutations and of rare codon usage is significantly lower for the replicase gene than for the protein VP1 coding region. PMID- 2993106 TI - Organisation and control of the Escherichia coli uvrC gene. AB - The Escherichia coli uvrC gene has been cloned into multicopy plasmids from the transducing phage lambda uvrC+ and the structural gene assigned to a 1.9-kb BglII fragment. Deletion of upstream sequences shows the presence of an in vivo uvrC promoter close to the start of the structural gene, as confirmed by subcloning the uvrC fragment into actively transcribed or 'promoter-free' restriction sites in various plasmid vectors. The control of uvrC transcription has been investigated using hybrid uvrC-cat operons. There are at least two promoters upstream of uvrC. Only the proximal promoter, some two orders of magnitude less effective than the cat promoter, is required for in vivo expression of the uvrC gene. We can find no evidence that expression of the uvrC gene on multicopy plasmids is either autogenously controlled or controlled by the product of the lexA gene. PMID- 2993107 TI - Human papillomavirus DNA in conjunctival papilloma. AB - A histologically proven conjunctival papilloma was examined for the presence of papillomavirus DNA by nucleic acid hybridization under relaxed conditions. It harbored papilloma-virus-specific DNA, which showed no detectable relationship with that of known virus types when tested under stringent conditions. This demonstrates that conjunctival papillomas are associated with more than one papillomavirus type. Viral diagnosis may turn out to be of prognostic value. PMID- 2993108 TI - Beta-adrenergic receptor subtypes iris-ciliary body of rabbits (1984,221:167-170) PMID- 2993109 TI - [Combined effect of organophosphate pesticides and ammonia released from fertilizers]. PMID- 2993110 TI - [Hygienic basis for a scale of hazards for the migration of agricultural chemicals from the soil into a surface water source]. PMID- 2993111 TI - Myelolactoferrin index (MLF) and myelopoiesis during bacterial infections and malignant diseases. PMID- 2993114 TI - Oncogenes of avian acute leukemia viruses are subsets of normal cellular genes. PMID- 2993113 TI - Chromatin structure of the human c-myc oncogene: definition of regulatory regions and changes in Burkitt's lymphomas. PMID- 2993112 TI - Modulation of gene expression through DNA-binding proteins: is there a regulatory code? PMID- 2993115 TI - Structure and function of ras and Blym oncogenes. PMID- 2993116 TI - TLym 1, a stage-specific transforming gene which shares homology to MHCI genes and encodes a secreted protein. PMID- 2993117 TI - The mil/raf and myc oncogenes: molecular cloning and in vitro mutagenesis. PMID- 2993118 TI - Characterization of the number of carbohydrate chains on the avian erythroblastosis virus erb B gene product and their role in transformation. PMID- 2993119 TI - Characterization of hematopoietic cells transformed in vitro by AEV-H, a v-erbB containing avian erythroblastosis virus. PMID- 2993120 TI - Different cellular substrates of Abelson leukemia virus transforming protein kinase in murine fibroblasts and lymphocytes. PMID- 2993121 TI - Regulin, a cytoskeleton-associated protein affecting phosphorylation dephosphorylation. PMID- 2993122 TI - Viruses as tumor initiators and tumor promoters. PMID- 2993123 TI - Endemic and sporadic cases of Epstein-Barr virus-positive Burkitt's lymphoma: immunological characterization of derived cell lines. PMID- 2993124 TI - Herpesvirus HVMA: a new representative in the group of the EBV-like B lymphotropic herpesviruses of primates. PMID- 2993125 TI - The human T-cell leukemia virus family, adult T cell leukemia, and AIDS. PMID- 2993127 TI - The molecular biology of human T cell leukemia virus and its roles in leukemogenesis of adult T cell leukemia. PMID- 2993128 TI - HTLV-I antibodies associated with cutaneous T cell lymphoma in Denmark. PMID- 2993126 TI - Molecular studies of human T cell leukemia/lymphotropic retroviruses. PMID- 2993129 TI - Presence of antibodies to the human T-cell leukemia virus HTLV I in German patients with symptoms of AIDS. PMID- 2993130 TI - Roles of HTLV-I p19 and natural antibody to HTLV-I in host immune responses. PMID- 2993131 TI - Antibodies to human T-cell leukemia virus-membrane antigens in macaques with malignant lymphoma. PMID- 2993132 TI - Coordinated expression of c-myc gene and a multigenic set may modulate the malignant phenotype in human haemopoietic cell lines. PMID- 2993133 TI - Molecular cloning of cDNA for the human interleukin-2 receptor. PMID- 2993134 TI - Are activated proto-onc genes cancer genes? PMID- 2993135 TI - Investigations on the effects of heavy water (D2O) on the functional activity of human platelets. AB - Heavy water (D2O) is able to activate or to inhibit biological systems and thus influences metabolism and functions of tissues and organs. On the basis of results obtained in preservation experiments performed on various organs, the effects of D2O on the functions of human platelets (retraction, spreading, aggregation induced by adrenaline, ADP and collagen) have been investigated. The effects of D2O, involving changes of membrane receptor conformation, binding strength of substances to their specific receptors, pH-values or enzyme activities, resulted in a significant decrease of ADP- and collagen-induced aggregation, while adrenaline-induced aggregation was significantly increased at the same D2O concentration. Retraction in the presence of D2O was significantly inhibited in a dose-related manner, while spreading was diminished, but not fully inhibited even by highest concentrations of D2O (95%). PMID- 2993136 TI - On the retraction of collagen and fibrin induced by normal, defective and modified platelets. AB - Fibrin clot retraction (FCR) and collagen gel retraction (CGR) were studied in patients with inherited platelet defects, i.e. in Glanzmann's thrombasthenia, Hermansky-Pudlak syndrome, May-Hegglin anomaly, giant platelet syndrome, as well as in patients with von Willebrand disease and factor XIII deficiency. FCR was abnormal only in thrombasthenia, while CGR was found to be reduced in 2 patients with Hermansky-Pudlak syndrome and in 4 out of 5 cases with von Willebrand disease. Both FCR and CGR were normal in May-Hegglin anomaly, giant platelet syndrome and severe factor XIII deficiency. In none of the examined bleeding disorders was a concomitant FCR and CGR reduction detected. Natural polyamines and anti-fibronectin antibodies did not affect platelet potency in FCR or CGR. Peroxidation of platelet membrane components by sodium periodate abolished the platelet-induced FCR and CGR. This effect was reversed by subsequent reduction by sodium borohydride. PMID- 2993137 TI - [Pericardial tamponade as a presenting symptom in cancer]. PMID- 2993138 TI - [Brain nicotinic acetylcholine receptors and their functions in special reference to central cardiovascular regulation]. AB - The nicotinic acetylcholine receptors present in the mammalian brain have received less attention than those in other areas of the nervous system. However, recent studies have reported the pharmacological character and regional distribution of a high-affinity binding of [3H]acetylcholine or [3H]nicotine. In this review, the pharmacological characters and physiological functions of the brain nicotinic receptors were discussed with special reference to central cardiovascular regulation. PMID- 2993140 TI - Comparison of the sensitivity of the vaginal mucous membranes of the albino rabbit and laboratory rat to nonoxynol-9. AB - Studies were conducted to compare the sensitivities of the vaginal mucous membranes of the albino rabbit and albino rat to irritation by the surfactant nonoxynol-9. The purpose of this comparison was to evaluate the rat as a model for screening vaginal products for their potential irritancy. Nonoxynol-9 at various concentrations in distilled water was administered intravaginally by lavage once a day for four consecutive days. On day 5, the animals were killed and the vaginal tissues were processed and examined histopathologically. Using a predetermined semiquantitative scoring system, it was found that the intensity of irritation was concentration dependent in both species. In the rabbit, the irritation was significantly greater than that in the rat, being mild with concentrations of 2.5 and 5% and moderate to severe at 12.5 and 25%. In the rat, irritation was negligible up to 12.5%, becoming mild at 25% and more moderate at 75%. Thus the vaginal mucosa of the rabbit is more sensitive than that of the rat and may be expected to give a more exaggerated representation of the irritation potential of a compound to the human vagina than would the rat mucosa. PMID- 2993139 TI - [The role of the pituitary in the development of electroacupuncture analgesia in rats]. AB - In order to investigate the involvement of pituitary beta-endorphin in electroacupuncture analgesia (EAA), the effects of hypophysectomy, dexamethasone (Dex) and adrenalectomy on the analgesia and the increase in plasma corticosterone (Cort) and ACTH levels produced by electroacupuncture (EA) were studied in male SD rats. In saline-treated and Dex-treated rats, plasma Cort levels were correlated with plasma ACTH levels. In non-treated rats, the time course of EA-induced increase in pain threshold was similar to that of EA-induced elevation of plasma Cort levels. In the hypophysectomized rats, EAA was significantly reduced and the EA-induced increase in plasma Cort was also abolished. Single administration of a large dose of Dex tended to reduce EAA and significantly reduced the EA-induced increase in plasma Cort and ACTH. Further suppression of pituitary functions by 4 days-treatment with Dex resulted in further reduction of EAA and the EA-induced increase in plasma Cort and ACTH. On the other hand, hind-paw pressure test without EA produced an increase in plasma Cort and ACTH to the same extent as that produced by EA and produced no analgesia. In the adrenalectomized rats, EAA was reduced, and the plasma ACTH level, which was sixteen times higher than that of nonoperated rats, was further elevated 2-fold higher by EA. No correlation between plasma ACTH levels and the increase in pain thresholds was observed in individual rats of the saline-treated and Dex-treated groups. Control pain thresholds were not influenced by hypophysectomy, Dex or adrenalectomy. These results suggest that pituitary beta endorphin may not be mainly involved in EAA. PMID- 2993141 TI - [Studies following implantation of hydroxyapatite ceramics in artificial cleft palate in the rat]. PMID- 2993142 TI - Indirect evidence against a contribution of the guanine nucleotide-binding inhibitory component of adenylate cyclase to impaired lipolysis in the epididymal adipose tissue of congenitally obese (ob/ob) mice. AB - The mixed adrenergic agonist epinephrine, at a 10 microM concentration, stimulated cyclic AMP production and glycerol release in the epididymal adipose tissue of ob/ob male mice. These effects when tested, respectively, after 7 min in the presence and after 60 min in the absence of theophylline were, however, 7- and 5-fold lower than in lean controls. The alpha-adrenergic blocker phentolamine and adenosine deaminase (which destroys extracellular adenosine) did not restore a normal lipolytic response to epinephrine in the adipose tissue of ob/ob mice. These data provide indirect evidence against a hyperactive mechanism in the coupling of alpha-adrenergic receptors and adenosine receptors to Ni, the guanine nucleotide-binding inhibitory component of adenylate cyclase, as the cause of reduced lipolysis in the adipose tissue of ob/ob mice. PMID- 2993143 TI - Metabolic and hormonal parameters after insulin-induced hypoglycemia in man, comparison between biosynthetic human insulin and purified pork insulin. AB - Although clinically undistinguishable, some authors have found important differences in the counterregulatory response between Biosynthetic Human Insulin (BHI) and Purified Pork Insulin (PPI). To reassess the problem 10 healthy volunteers of both sexes underwent paired iv insulin tolerance test with both BHI and PPI (0.10 U/kg b.w.). To check the humoral response the variations of glucose, free fatty acids (FFA), prolactin, growth hormone, ACTH and plasma renin activity were evaluated. Blood glucose depression and further recovery by BHI and PPI administration paralleled each other, so were, prolactin, FFA, and plasma renin activity. A slight section of ACTH, and GH was observed under BHI challenge. There were not statistically significant differences between both insulins on any of the six parameters studied. The data do not confirm earlier published reports indicating hormonal and metabolic differences between human and porcine insulin. PMID- 2993144 TI - Effects of naloxone-induced opiate receptors blockade on insulin secretion in obesity. PMID- 2993145 TI - Acetaminophen hepatotoxicity and targeted rescue: a model for specific chemotherapy of hepatocellular carcinoma. AB - We have taken advantage of the presence of hepatic receptors for galactose terminal (asialo-)glycoproteins to achieve targeted rescue of differentiated hepatocytes from acetaminophen-induced toxicity in vitro. To accomplish this, a conjugate was formed by covalent coupling of N-acetylcysteine (an acetaminophen antagonist) to galactose-terminal (asialo-)fetuin. We used two human hepatocyte derived cell lines to test our targeted-rescue method: Hep G2 cells are capable of receptor-mediated endocytosis of galactose-terminal glycoproteins and PLC/PRF/5 cells are not. In the presence of acetaminophen alone, both cell lines demonstrated a similar concentration-dependent sensitivity. Growth rates of both cell lines became normal when N-acetylcysteine was administered in equimolar quantities with acetaminophen indicating that both cell lines had the potential of responding to the antagonist. When asialofetuin-N-acetylcysteine conjugate was given to both cell lines in the presence of acetaminophen, PLC/PRF/5, receptor ( ) cells failed to respond. However, Hep G2, receptor (+) cells treated with asialofetuin-N-acetylcysteine conjugate under identical conditions, increased their populations and eventually reached confluence. Control conjugate fetuin-N acetylcysteine as well as asialofetuin alone had no effect on either cell line. PMID- 2993146 TI - Hepatocellular carcinoma in young, mentally retarded HBsAg carriers without cirrhosis. AB - In a Californian institution for the mentally retarded, surveillance by autopsy of all deaths (n = 138) identified three cases of hepatocellular carcinoma. Hepatocellular carcinoma cases occurred in young (mean age = 26 years) male carriers of HBsAg and were not associated with cirrhosis. They were of the nonfibrolamellar oncocytic type and were rapidly fatal. Hepatocellular carcinoma incidence in HBsAg carriers was estimated to be 246 times greater than United States males. PMID- 2993147 TI - Improved survival following injection sclerotherapy for esophageal varices: final analysis of a controlled trial. AB - Long-term follow-up (median: 37 months; range: 19 to 68) of the 116 patients (56 sclerotherapy, 60 control group) entered into a controlled trial of endoscopic variceal sclerotherapy has shown a total of 18 deaths in the sclerotherapy group, including five from variceal bleeding compared with 32 deaths in the control group (p less than 0.01), of which 25 were from variceal hemorrhage (p less than 0.001). Survival as assessed by cumulative life analysis was significantly better in those treated by sclerotherapy (p less than 0.001). Both the cumulative proportion of patients rebleeding and the total number of episodes of variceal hemorrhage were also significantly less in the sclerotherapy group (p less than 0.01). Recurrence of varices was observed in 27 of 45 patients in whom variceal obliteration was initially observed at a median of 11 months (range: 2 to 27) later, although in only 12 of these did bleeding recur and was the cause of death in one. PMID- 2993148 TI - Prognostic value of thyroid hormone levels in patients evaluated for liver transplantation. AB - The thyroid hormones T4, T3, rT3 and TSH were assayed in 134 adult patients evaluated and accepted as potential liver transplant candidates at the University of Pittsburgh from March, 1981 to December, 1983. The subsequent course of these patients was evaluated with respect to the levels of these hormones obtained at the time of acceptance for transplantation. T4 levels were increased significantly while their T3 levels were reduced (both p less than 0.01) in those who survived and were discharged home as compared to either those who died waiting to be transplanted or died following the procedure. As a result, the ratio of T3/T4 was reduced markedly (p less than 0.01) in those who were transplanted and survived as compared to those not transplanted or dying following transplantation. Importantly, the rT3 levels clearly separated (p less than 0.01) those who would die prior to transplantation from those who would survive to be transplanted. Finally, the ratio rT3/T3 even more clearly separates those who will die prior to transplantation (p less than 0.01) from the other two groups. These data suggest that thyroid hormone levels, particularly rT3 levels, might be useful in setting priorities for which patients referred for a transplantation evaluation should be accepted into the program and in determining who among accepted patients should be operated upon in preference to others also accepted and waiting to be transplanted. PMID- 2993149 TI - Transferrin and its receptor: their roles in cell function. AB - Transferrin, its receptor and the entry of iron into the cell have sprung into prominence because of recent evidence that proliferation of various cell types involves regulation of this sequence of events, as evidenced especially by changes in receptor number. A third component functionally linked to transferrin and its receptor is the intracellular iron-storage protein, ferritin, which ensures against toxic levels of free ferrous iron, which might otherwise cause peroxidative damage to cell membranes and other cell structures (1). In this article, we shall focus on interactions between these three proteins of iron exchange, their roles in homeostasis and especially their role in relation to the liver which is a major organ of iron storage. PMID- 2993151 TI - Ultrastructural similarities of adenoid cystic carcinoma and pleomorphic adenoma. AB - Ultrastructural examination of five adenoid cystic carcinomas, three breast and two salivary gland, reveals identical patterns of tumour cell differentiation, organization and distribution of cellular products (Zaloudek, Oertel & Orenstein 1984). In both sites, there is proliferation of two populations of cells, one with characteristics and organization of duct-type luminal epithelial cells and a second that forms the principal proliferating component and has the overall organization and appearance that would suggest that they represent modified myoepithelial cells. Recent ultrastructural studies also indicate that tumour cell types and histological organization similar to those described for adenoid cystic carcinoma occur during histodifferentiation of salivary gland pleomorphic adenoma (Dardick et al. 1983a, b). The characteristic histological pattern of adenoid cystic carcinoma is dependent on the formation of pseudolumina containing proteoglycans and reduplicated basal lamina. Similar, but smaller, lumina of like organization and contents are evident in some cases of pleomorphic adenoma. Both the ultrastructural similarities of the tumour cell types and their organization, in adenoid cystic carcinoma and pleomorphic adenoma, suggest that these tumours have a similar histogenetic basis. The fact that one lesion is malignant and the other benign does not preclude common types of tumour cells and developmental processes. PMID- 2993150 TI - Receptor-mediated endocytosis and targeted drug delivery. PMID- 2993152 TI - Pathobiologic study of pituitary tumors: report of 62 cases with a review of the recent literature. AB - Advances in radioimmunoassay procedures, immunocytochemistry, neuroradiologic imaging, and the surgical and medical treatment of pituitary adenomas have led to reappraisal of their classification as well as refinements in the diagnostic approaches used by pathologists. Sixty-two pituitary adenomas are described, and recent advances in this field are reviewed. Most of the patients were adults, but one of the adrenocorticotropic hormone (ACTH)-producing adenomas occurred in an 11-month-old infant. Endocrine-inactive tumors (43.5 per cent) were less common than hormone-producing tumors (56.5 per cent). Local invasion was most common in the former group, followed by ACTH-producing and other hormone-producing tumors. Ultrastructural features correlated with hormonal levels in the growth hormone(GH)-secreting tumors but not in the prolactin(LTH)- or ACTH-producing tumors. The formation of 7-nm filaments in the cytoplasm of tumor cells, corresponding to Crooke's hyaline change on light microscopy, was characteristic of ACTH-producing tumors. Ultrastructural changes in the ACTH granules suggested that the filaments may be derived from the feedback action of cortisol. Prior to surgery, a Rathke's cleft cyst and a chordoma were mistaken for endocrine inactive pituitary adenomas. In two additional cases ectopic ACTH-producing tumors of lung clinically mimicked pituitary adenoma. Conversely, one pituitary adenoma mimicked sphenoid wing meningioma. Clinical, hormonal, and radiologic data and immunocytochemical and electron microscopic studies are needed for accurate pathologic interpretation and classification of pituitary adenomas. PMID- 2993153 TI - Epithelial membrane antigen--a diagnostic discriminant in surgical pathology: immunohistochemical profile in epithelial, mesenchymal, and hematopoietic neoplasms using paraffin sections and monoclonal antibodies. AB - Glycoproteins isolated from human milk fat globule membranes, designated epithelial membrane antigen (EMA), have been detected immunohistochemically in most nonneoplastic epithelia and are potentially a highly effective marker for establishing the epithelial nature of neoplastic cells. With commercially available monoclonal antibodies and an indirect immunoperoxidase technique, EMA localization was evaluated in paraffin-embedded tissues from a wide variety of neoplasms (320 specimens). Adenocarcinomas from various primary sites (breast, lung, colon, stomach, pancreas, gallbladder, prostate, endocrine glands, ovary, kidney, thyroid) were immunoreactive for EMA in 88 of 97 cases (91 per cent). Cytoplasmic and apical luminal membrane staining were the most common patterns of immunoreactivity, with peripheral membrane staining or other patterns also seen in some neoplasms. Squamous cell (13 of 13 cases) and transitional cell (12 of 12 cases) carcinomas, small cell anaplastic carcinomas (12 of 12 cases), and mesotheliomas (six of six cases) were also uniformly EMA-positive. Malignant lymphomas of the Hodgkin's (15 cases) and non-Hodgkin's types (74 cases), except for the true histiocytic lymphomas and occasional T-cell lymphomas, were nonreactive for EMA. Neoplastic and nonneoplastic plasma cells showed variable EMA positivity. Endocrine neoplasms (17 cases), including carcinoid tumors, medullary carcinoma of thyroid, adrenocortical carcinomas and pheochromocytomas, and germ cell tumors (eight cases, embryonal carcinoma and seminoma), and a wide variety of soft tissue tumors (27 cases) generally lacked immunoreactivity for EMA; the exceptions to this finding were synovial sarcomas and an epithelioid sarcoma. Malignant melanomas (eight cases) were typically nonreactive. Based on the observations in this large series of neoplasms, EMA is an excellent marker of epithelial differentiation, appears to be highly reliable for discriminating between poorly differentiated carcinomas and malignant lymphomas, and is especially helpful in characterizing small cell anaplastic carcinomas. Epithelial membrane antigen immunoreactivity is well preserved in paraffin sections of routinely processed tissues, facilitating application of this technique in diagnostic surgical pathology. PMID- 2993155 TI - Definitive localization of Becker muscular dystrophy in Xp by linkage to a cluster of DNA polymorphisms (DXS43 and DXS9). AB - A study of linkage between Becker muscular dystrophy and four X chromosome specific DNA polymorphisms in 17 kindreds has indicated that this gene is located in Xp, as already anticipated by single pedigree analysis. In particular the DXS43 and DXS9 loci, identified by probes D2 and RC8, respectively, are closely linked to each other and are both located at approximately 15 cM from the Becker locus. These linkage data, together with the previously established linkage between Becker and the DXS7 locus identified by probe L 1.28, indicate that the Becker gene is located in the same region where Duchenne has been mapped and also yield information about relative genetic distances among different DNA polymorphisms of the X chromosome. PMID- 2993154 TI - Genetic linkage heterogeneity in the fragile X syndrome. AB - Genetic linkage between a factor IX DNA restriction fragment length polymorphism (RFLP) and the fragile X chromosome marker was analyzed in eight fragile X pedigrees and compared to eight previously reported pedigrees. A large pedigree with apparently full penetrance in all male members showed a high frequency of recombination. A lod score of -7.39 at theta = 0 and a maximum score of 0.26 at theta = 0.32 were calculated. A second large pedigree with a nonpenetrant male showed tight linkage with a maximum lod score of 3.13 at theta = 0, a result similar to one large pedigree with a nonpenetrant male previously reported. The differences in lod scores seen in these large pedigrees suggested there was genetic heterogeneity in linkage between families which appeared to relate to the presence of nonpenetrant males. The combined lod score for the three pedigrees with nonpenetrant males was 6.84 at theta = 0. For the 13 other pedigrees without nonpenetrant males the combined lod score was -21.81 at theta = 0, with a peak of 0.98 at theta = 0.28. When lod scores from all 16 families were combined, the value was -15.14 at theta = 0 and the overall maximum was 5.13 at theta = 0.17. To determine whether genetic heterogeneity was present, three statistical tests for heterogeneity were employed. First, a "predivided-sample" test was used. The 16 pedigrees were divided into two classes, NP and P, based upon whether or not any nonpenetrant males were detected in the pedigree. This test gave evidence for significant genetic heterogeneity whether the three large pedigrees with seven or more informative males (P less than 0.005), the eight pedigrees with three informative males (P less than 0.001), or all 16 pedigrees (P less than 0.001) were included in the analysis. Second, Morton's large sample test was employed. Significant heterogeneity was present when the analysis was restricted to the three large pedigrees (P less than 0.025), or to the eight pedigrees with informative males (P less than 0.05) but not when smaller, less informative pedigrees were also included. Third, an "admixture" test for heterogeneity was employed which tests for linkage versus no linkage. A trend toward significance was seen (0.05 less than P less than 0.10) which increased when the analysis was restricted to the larger, more informative pedigrees.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993157 TI - Molecular evidence of triplication in the haptoglobin Johnson variant gene. AB - The protein and gene structure of the Hp Johnson variant (Hp3) were analyzed in two related heterozygous individuals. The molecular weight (23 kd) and amino acid composition of Hp3 alpha chain were in agreement with the triplicated structure first suggested by Smithies in 1964. Direct gene analysis by Southern blotting showed a three-fold tandem repeat of the same 1.7 kb DNA segment implicated in the Hp2 gene duplication. On the basis of these data a nine exon model for the Hp3 gene is proposed. PMID- 2993156 TI - An EcoRI restriction fragment length polymorphism (RFLP) in the human c-erb A locus. AB - An EcoRI restriction fragment length polymorphism (RFLP) was detected in the 3' end of the locus of the c-erb-A proto-oncogene. The frequency of the rarer allele was around 3.0% in a normal population of 107 unrelated individuals. This frequency did not significantly differ in DNA samples from patients with breast tumors or acute leukemias. PMID- 2993158 TI - Genetic linkage relationships of seven DNA probes with Duchenne and Becker muscular dystrophy. AB - The inheritance of seven restriction fragment length polymorphisms detected by DNA probes has been studied in families with Duchenne and Becker muscular dystrophies (DMD and BMD). The probes used have all been mapped to the short arm of the X-chromosome, four being distal and three proximal to the disease loci located within the Xp21 region. Linkage analysis of the DNA polymorphisms in relation to the two disorders showed similar genetic distances. Data obtained from DMD and BMD families have been combined to give more precise values for the different recombination fractions. Combined use of these polymorphic DNA markers will be of practical value in the genetic counselling of women at risk for Duchenne and Becker muscular dystrophy. PMID- 2993159 TI - Identification of a deletion in the low density lipoprotein (LDL) receptor gene in a patient with familial hypercholesterolaemia. AB - DNA samples from 60 unrelated UK patients with familial hypercholesterolaemia (FH) were screened by Southern blot hybridisation to detect gross alterations in the low density lipoprotein (LDL) receptor gene. One patient was found to have a 2kb deletion in the 3' part of the gene. The deletion cosegregates with the FH phenotype in his family. This finding is compatible with the deletion being the cause of FH in this case and makes a presymptomatic test based on DNA analysis available for this family. The defects in most of the other patients are likely to be due to point mutations. PMID- 2993160 TI - Predicted secondary structure and hydrophobic cores of T-cell (TCGF or IL-2) and platelet-derived growth factors as well as of transforming protein p28sis of simian sarcoma virus are similar to those of interferons. AB - Estimation of a content of secondary structures from amino acid composition of IL 2 (TCGF), PDGF and transforming protein p28sis of simian sarcoma virus by means of the previously suggested nomograms (V.P. Zav'yalov and A.I. Denesyuk (1982) Immunol. Lett. 4, 7-14) allowed us to conclude that a dominant secondary structure of these proteins should be alpha-helix. Its content in PDGF and p28sis was estimated to be approximately equal to 50%, and that in IL-2 to be 50-65%. The content of beta-pleated structure in PDGF and p28sis does not exceed 10%, and that in IL-2 is found to be 20-30%. In all the proteins investigated five alpha helical segments can be distinguished, of which one of the surfaces contains clusters of bulky hydrophobic side chains. As the number of alpha-helical segments satisfying the principles of packing into a hydrophobic core as well as the overall length of a polypeptide chain of IL-2 and PDGF coincided with those for IFNs, it was decided to arrange alpha-helices of these proteins into a globular structure by analogy with IFNs (V.P. Zav'yalov and A.I. Denesyuk (1984) Doklady Akad Nauk S.S.S.R. 275, 242-246). In consequence, identical side chains of amino acid residues were found at 9 out of 29 positions in hydrophobic cores of IFN-beta and IL-2, p28sis (PDGF), respectively. Thus the homology of hydrophobic cores of the proteins compared is greater than 30%. Probability of chance coincidences of amino acid residues in the hydrophobic cores of p28sis (PDGF) and IFN-beta is found to be 0.03, and that deduced from comparison of IL-2 and IFN-beta is 0.05.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993162 TI - Murine invariant chain gene: chromosomal assignment and segregation in recombinant inbred strains. PMID- 2993163 TI - Demonstration of a ouabainlike plasma compound in hypertension prone and hypertension resistant rats. AB - Material extracted and partially purified from plasma of the Sabra hypertension prone rats was found to be capable of 1) inhibiting the binding of 3H-ouabain to rat brain synaptosomes, 2) inhibiting the activity of rat brain microsomal Na, K activated adenosine triphosphatase, and 3) increasing the contractile force of rat heart muscle. The results demonstrate the presence of a ouabainlike compound in the plasma of these rats. The plasma concentration of this compound in Sabra hypertension prone rats was 698 +/- 199 nmol/ml in ouabain equivalents (SEM; n = 11) versus 2543 +/- 1140 nmol/ml (n = 9) in the Sabra normotensive strain. The presence of ouabainlike compound in the plasma is consistent with the hypothesis that this compound functions as a hormone that regulates Na, K activated adenosine triphosphatase activity and the physiological processes in which this enzyme is involved. PMID- 2993161 TI - Structure and polymorphism of class I MHC antigen mRNA. AB - We have used cDNA cloning and primer extension techniques to determine the complete nucleotide sequence of HLA-B7 mRNA. The 5'-untranslated sequence of the mRNA is rather short and the putative promoter has weak homology to the conventional "TATA" sequences. The 5' end and the polyadenylation site define the transcription unit of the B7 gene to be about 3.5 kb long. Comparison of the translated nucleotide sequence of this cDNA with the amino acid sequence of the heavy chain of the B7 antigen showed two amino acid differences. In addition, comparison with the sequences of the coding and untranslated regions of several HLA and H-2 genes showed that the class I histocompatibility molecules consist of four variable segments separated by three regions of homology. Analysis of the DNA polymorphisms revealed that in the variable segments the majority of the nucleotide substitutions are nonsilent, while in the homology regions the majority of substitutions are silent. Further analysis of the nature of the amino acid substitutions revealed the predominance of nonconservative replacements/changes in both the variable segments and the homology regions except the transmembrane part of the molecule. Selection at both the protein and the codon levels contributes to the pattern of mutations found in class I histocompatibility molecules. PMID- 2993164 TI - Cardiac beta-adrenergic receptors in salt-dependent genetic hypertension. AB - The pattern of cardiac beta-adrenergic receptor changes in different hypertrophy models varies according to the pathophysiology. In salt-sensitive Dahl rats, high dietary salt intake leads to a moderate degree of cardiac hypertrophy associated with increased numbers of cardiac beta-adrenergic receptors but unchanged affinity for agonists. Isoproterenol-stimulated cardiac adenylate cyclase is also higher in salt-loaded hypertensive rats without any change in basal or NaF stimulated activities. In contrast, neither beta-adrenergic receptors nor adenylate cyclase activities are affected by variations in dietary salt in salt resistant Dahl rats. The extent of isoproterenol-induced down regulation of beta adrenergic receptors on isolated cardiac myocytes as well as the recovery from this down regulation is not significantly different in either strain of Dahl rats and is not influenced by dietary salt. The enhancement of beta-adrenergic pathways in salt-dependent genetic hypertension may be involved both in the initiation of cardiac hypertrophy and the preservation of contractile function. PMID- 2993166 TI - Malignant fibrous histiocytoma of the lung. PMID- 2993165 TI - Evidence for endothelium-derived relaxing factor in cultured cells. AB - Intracellular cyclic GMP concentration was used as a biochemical indicator of endothelium-dependent and organonitrate-induced responses to these vasodilators in cultured porcine aortic smooth muscle and endothelial cells. Sodium nitroprusside (10(-6) M) caused a rapid increase in cyclic GMP levels in confluent smooth muscle cell cultures but not in confluent endothelial monolayers. Adenosine triphosphate (10(-4) M) and methacholine (10(-5) M), two agents that elicit endothelium-dependent relaxation in intact vessels, failed to raise cyclic GMP concentrations in muscle or endothelial cultures alone. When the cell types were grown together in mixed culture, however, treatment with adenosine triphosphate or methacholine induced an elevation in intracellular cyclic GMP levels. These findings suggest that mixed cultures of arterial smooth muscle and endothelial cells can be used to study the phenomenon of endothelium dependent responses in arterial smooth muscle. PMID- 2993167 TI - DNA relationships among some tox-bearing corynebacteriophages. AB - The DNA genomes of a number of tox-bearing, temperate corynebacteriophages isolated from strains of Corynebacterium diphtheriae and Corynebacterium ulcerans were compared. With one exception, these phages displayed similarities in their restriction enzyme digest profiles and extensive homology with prototypic beta converting phage. The exception, phage delta, had a unique restriction profile and exhibited homology with beta over a limited portion of its genome. DNAs of phages from each host contained cohesive ends and integrated as prophage by a mechanism analogous to that employed by coliphage lambda. It is proposed that these tox-bearing phages belong to a common family, the beta family. The role of the beta family in the movement of the tox gene between strains of C. diphtheriae and C. ulcerans is discussed. PMID- 2993169 TI - Feline leukaemia virus. PMID- 2993168 TI - Effect of theophylline on differentiation of Trypanosoma brucei. AB - Differentiation of Trypanosoma brucei in the mammal limits the degree of parasitemia and prepares the trypanosome for passage back into the tsetse fly. In an attempt to define the signals that control differentiation, we found that theophylline, in contrast to indomethacin, blocked differentiation, prolonged parasitemia, elevated prostaglandin and cyclic AMP concentrations of rat plasma, and depressed intratrypanosomal cyclic AMP. Relatively nontoxic drugs that alter differentiation are powerful tools for elucidating the events that control this important process. PMID- 2993170 TI - Immunoglobulin production in Epstein-Barr virus-transformed lymphoblastoid cell lines in patients with ataxia telangiectasia. AB - Peripheral blood lymphocytes from 7 patients with ataxia telangiectasia were transformed by Epstein-Barr virus, and lymphoblastoid cell lines (LCL) were established. Immunoglobulin levels in the supernatants of the LCL were measured in order to analyze immunological abnormalities, especially the low levels of serum IgA in these patients. We found that the geometric means of IgA production in the supernatants of the LCL from the 7 patients were significantly lower than those from 9 healthy controls (5.0 vs. 834 ng/ml, p less than 0.001). These results indicate that the low levels of serum IgA in ataxia telangiectasia are attributable at least in part to an intrinsic abnormality of B cells. PMID- 2993171 TI - Radiosynthesis of [18F]3-acetylcyclofoxy: a high affinity opiate antagonist. AB - A convenient method for the preparation of high specific activity [18F]3 acetylcyclofoxy (3-acetyl-6-deoxy-6-beta-18F-fluoronaltrexone) was developed. The method utilizes reactor-produced [18F]-fluoride as its tetraethylammonium (TEA X F) salt in a SN2 displacement on a secondary triflate precursor. Typically, 45% of the 18F activity can be converted to the reactive TEAF in a 70 min preparation. From this, 35% yield (decay corrected) of the [18F]3-acetylcyclofoxy was obtained after HPLC purification with a specific activity of 25 Ci/mmol in a total synthesis time of 60 min. PMID- 2993172 TI - Serological survey of antibodies to the adult T-cell leukemia virus-associated antigen (HTLV-A) in Taiwan. AB - A serological survey of antibodies to the adult T-cell leukemia virus-associated antigen (HTLV-A) conducted in Taiwan revealed II seropositive individuals among 2,565 serum donors tested, which included 1,187 hospital or free clinic patients, 1,065 blood donors and 313 aborigines. The overall prevalence rate was 0.43% and was 0.90% (9/995) for adults aged 40 years or over, which was similar to the rates observed in the non-endemic areas in Japan. No highly prevalent area was demonstrated. The II seropositive individuals identified were 8 hospital or clinic patients, 1 blood donor and 2 aborigines. Among 108 various leukemia/lymphoma patients also studied, 7 were positive. These 7 anti-HTLV-A positive patients also had clinicopathologic features of adult T-cell leukemia/lymphoma (ATL). Among 21 relatives of 3 seropositive individuals in the epidemiologic study, 2 were positive; and among 25 relatives of 5 ATL patients, 6 were positive. Our data support the contention that horizontal familial transmission of human T-cell leukemia virus (HTLV-I) is an important route and HTLV-I infection can occur on a different genetic background. Furthermore, the antibody titer did not correlate with the manifestation or the state of the disease. PMID- 2993173 TI - Type-I and type-III HTLV antibodies in hospitalized and out-patient Zairians. AB - Sera from 182 Zairians (99 females and 83 males), aged 5 to 71 years, including maternity and child care consultants, out-patients suffering from minor injuries and patients hospitalized for tuberculosis, malaria or trauma, were analyzed for specific antibodies to HTLV-I and HTLV-III. Following pre-screening by the ELISA technique, reactive sera were further analyzed for specificity to HTLV-I or HTLV III antigens by competition and/or Western blotting experiments. African sera, possibly because they have higher immunoglobulin levels than US and European sera, are highly reactive in ELISA systems and confirmatory assays are essential to rule out false-positive results. Confirmed antibody prevalence for HTLV-I was 13.2% (II females and 13 males) and increased with age, suggesting continuous exposure to the virus throughout life. Confirmed antibody prevalence for HTLV-III was 6.0% (8 females and 3 males) and showed a peak age range between 21 and 40 years, suggesting heterosexual transmission. Individuals positive for HTLV-I antibodies were not the same as individuals positive for HTLV-III antibodies, suggesting that infection with one virus did not increase susceptibility to infection by the other virus. Further investigations of the epidemiology and of the immunovirology of HTLV-III in Zaire, in relation to acquired immuno deficiency syndrome (AIDS)-associated pathologies, should enlighten the question of the significant percentage of HTLV-III-infected individuals who do not manifest symptoms of AIDS. PMID- 2993174 TI - Mr 75,000 protein, a tumor marker in renal adenocarcinoma, reacting with antibodies to a synthetic peptide based on a cloned human endogenous retroviral nucleotide sequence. AB - Using a rabbit antiserum to a synthetic undecapeptide deduced from a cloned human retroviral gag-gene-related DNA sequence, we found a specific immunohistochemical reaction in all of 42 tested renal cell adenocarcinomas (RCC), while none of 17 similarly tested Wilms' tumors and 65 carcinomas at other sites were positive. The RCC included two cases that presented with distant metastases. It had not been possible to establish the origin of these until immunohistochemical staining revealed this typical reaction. Subsequent renal angiography disclosed the primary. In immunoblotting the antiserum detected an Mr 75,000 protein in RCC tissue, and this reaction was blocked by the undecapeptide. The usefulness of this protein as a tumor marker for RCC is discussed. PMID- 2993175 TI - The clinical features and diagnosis of a discordant atrioventricular connexion. AB - Seventy-three patients were studied with a discordant atrioventricular connexion (ages 3 months to 46 years). In 58 the diagnosis was proved by angiography and in the other 15 at necropsy. Forty-eight had usual atrial arrangement and 25 mirror image atria. Fifty-two patients had ventriculo-arterial discordance, 13 double outlet right ventricle, 4 ventriculoarterial concordance, 3 single outlet of the heart and 1 double outlet left ventricle. Nine with ventriculoarterial discordance had no other associated defects. The spatial relationship of the ventricles was variable, but in usual atrial arrangement the morphologically left ventricle tended to be on the right, and in mirror-image arrangement to the left of the morphologically right ventricle. The spatial relationship between the arterial valves was also variable, but in ventriculoarterial discordance and double outlet right ventricle the aortic valve was anterior and either to the right or left of the pulmonary valve in all but 3 cases. There is no typical clinical picture for these malformations. Symptoms depend upon the associated anomalies or the presence of atrioventricular dissociation. The diagnosis of a discordant atrioventricular connexion is best achieved by the echocardiographic and haemodynamic studies, but the electrocardiogram, chest radiograph and nuclear medicine studies may suggest its presence. Most patients are asymptomatic and progress normally to adult life. Only the symptomatic patients require surgical correction. Postoperative follow-up in the survivors is excellent, only 2 of 14 patients dying after surgery. PMID- 2993176 TI - Leukotrienes on porcine hemodynamics and prostanoid release. AB - We studied the effect of intracoronary leukotriene B4, C4, D4 and E4 (0.1-3 micrograms) on coronary artery blood flow and resistance in anesthetized pigs. Conventional hemodynamics were measured, and the peripheral electrocardiogram was obtained in lead II. Thromboxane B2 and 6-keto-prostaglandin F1 alpha (as breakdown products of thromboxane and prostacyclin, respectively) were measured during the influence of leukotrienes on the heart. All leukotrienes except B4 reduced coronary flow. Peak reduction was produced by 3 micrograms of each eicosanoid: C4 = 96 +/- 4%+; D4 = 98 +/- 2%+; E4 = 82 +/- 8%+. Coronary resistance increased after the same dose B4 = 65 +/- 18%; C4 = 225 +/- 94% (P less than 0.01); D4 = 442 +/- 118%+; E4 = 110 +/- 43% (+ = P less than 0.001). Increase in filling pressure and heart rate but blood pressure reduction and diminution in left ventricular d P/dtmax were observed with leukotriene C4, D4 and E4. The S-T segments of the electrocardiogram were elevated, thus indicating myocardial ischemia during the blood flow reduction. Indomethacin (5 mg/kg i.v.) had no effects on the leukotriene-induced hemodynamic sequelae. Thromboxane B2 concentration in coronary sinus blood plasma increased by 132-176% (P less than 0.05) at peak leukotriene effects on blood flow. Thus, leukotriene C4, D4, and E4 are vasoconstrictors in the situ porcine heart. Leukotriene B4, however, exerts no hemodynamic effects. The electrocardiographic ischemia and changes in hemodynamics indicate actions on coronary resistance and myocardial depression. These eicosanoids may contribute to cardiac dysfunction and vasospasm in coronary artery disease. PMID- 2993178 TI - Unusual intracardiac metastasis of a testicular embryonal carcinoma. AB - In this report, we present a case of a metastasis of an embryonal carcinoma, unusually located in the right atrium, which was clinically revealed by a systemic paradox embolism through an atrial septal defect. The primary tumor was found in an undescended testis. PMID- 2993179 TI - Anti-opiate (naloxone) suppression of Cushingoid degenerative changes in obese/SHR. AB - Young and mature, genetically obese and non-obese, spontaneously hypertensive rats (SHR) were injected with saline (controls) or naloxone for 12 weeks. Naloxone stilled the hyperphagia to a normal intake in the obese SHR (Obese/SHR) so that young Obese/SHR did not develop their usual massive obesity and mature Obese/SHR that had become massively obese were reduced to leanness. The naloxone treated young, obese and non-obese SHR (controls) exhibited marked reduction of the weight of their pituitary and adrenal glands, whereas the pituitary and adrenal glands of naloxone-treated mature, obese and non-obese/SHR were greatly increased in weight. The elevated systolic blood pressure of the obese and non obese rats was reduced after chronic treatment with naloxone. Naloxone treatment caused reduction of blood ACTH, corticosterone, and beta endorphin levels but elevated growth hormone levels. The characteristic hyperinsulinemia, hyperlipidemia, hyperglycemia, elevated BUN levels, and the Cushingoid spectrum of degenerative changes found in Obese/SHR did not appear in naloxone-treated rats. PMID- 2993180 TI - Effects of antiserum to Trypanosoma cruzi on the uptake and rate of killing of vector-borne, metacyclic forms of the parasite by macrophages. PMID- 2993182 TI - Synthesis and biological activity of linear and cyclic enkephalins modified at the Gly3-Phe4 amide bond. AB - As part of our continuing effort to define structure-activity relationships for enkephalin and design enzymatically resistant analogs, we report the synthesis and biological activities of linear and cyclic enkephalin analogs modified at the Gly3-Phe4 amide bond. The partial retro-inverso enkephalin analog Tyr-D-Ala-gGly (R,S)-mPhe-Leu-NH2 and its cyclic counterpart, Tyr-cyclo[D-A2 bu-gGly-(R,S)-mPhe Leu-], were synthesized as diastereomeric mixtures using solution methodology. The racemic benzylmalonate allowed the linear analog to be synthesized by fragment coupling at the reversed bond. Cyclization of the second analog was carried out at high concentration, eliminating formation of polymer by the use of an insoluble base. All gem-diaminoalkyl residues were prepared by conversion of peptidyl amides with benzene iodonium bis(trifluoroacetate). Diastereomers of both compounds were separable by reverse phase HPLC but those of the linear compound racemized rapidly under conditions of testing and were therefore tested together. All analogs tested had activities ranging from 6 to 14% of the activity of Leu enkephalin, indicating that the Gly3-Phe4 amide bond is important, though not crucial, for receptor binding. PMID- 2993177 TI - Radionuclide diagnosis of anomalous origin of the right pulmonary artery from the ascending aorta (so-called hemitruncus). AB - A 5-week-old infant presented with signs of severe congestive heart failure and pulmonary hypertension. Injection of technetium-99m pertechnetate demonstrated anomalous perfusion of the right lung. Subsequently, anomalous origin of the right pulmonary artery from the ascending aorta was proven at cardiac catheterization and repaired. Repeat injection of radionuclide 1 week postoperatively demonstrated normal flow to the right lung. PMID- 2993181 TI - Beta-endorphin. Synthesis and biological activity of analogs with disulfide bridges. AB - Two analogs of human beta-endorphin (beta-EP) which contain cystine bridges, [Cys15-Cys26,Phe27,Gly31]-beta-EP (I) and [Cys16-Cys26,Phe27,Gly31]-beta-EP (II), were synthesized by the solid-phase method. Peptides I and II were shown to contain 2-2.5 times the opiate receptor binding activity of beta-endorphin. We also synthesized two analogs with reduced alkylated cysteine residues and these peptides, [Arg9,19,24,28,29 Cys(Cam)11,26,Phe27,Gly31] and [Arg9,19,24,28,29,Cys (Cam)12,26,Phe27,Gly31], were shown to have approximately the same opiate receptor activity as beta-endorphin. PMID- 2993183 TI - Model studies for the direct effect of high-energy irradiation on DNA. Mechanism of strand break formation induced by laser photoionization of poly U in aqueous solution. AB - Laser flash photolysis of polyuridylic acid (poly U) in anoxic aqueous solutions leads to biphotonic photoionization of the uracil moiety followed by the formation of single strand breaks (ssb). The rate constant for ssb formation (1.0 s-1, obtained from the slow component of conductivity increase at 23 degrees C and pH 6.8) increases with decreasing pH to 235 s-1 at pH 3.5. The activation energy (pre-exponential factor) was measured to be 66 kJ mol-1 (5 X 10(11) s-1) at pH 6.8. Addition of dithiothreitol (DTT) or glutathione (GSH) prevents ssb formation by reacting with a poly U intermediate (rate constant = 1.2 X 10(6) and 0.16 X 10(6) dm3 mol-1 s-1, respectively). Since with OH radicals as initiators very similar data have been obtained for the kinetics of ssb formation and for the reaction with DTT, we conclude that photoionization of the uracil moiety in poly U leads eventually to the same chemical pathway for ssb formation as that induced by OH radicals. Furthermore, we propose that protection by DTT and GSH occurs via H donation to the C-4' radicals of the sugar moiety of DNA and to the C-4' and the C-2' radicals of poly U. PMID- 2993184 TI - Otorhinolaryngologic disorders of adolescents: a review. PMID- 2993185 TI - Effects of leukotrienes on the electroretinographic profile of the isolated rat retina. AB - The isolated retina represents an excellent membrane model since it provides a specific electrophysiological response that is easy to quantify. Therefore the authors have tested the effects of B4, C4 and D4 leukotrienes on this model. In the presence of leukotrienes, electrical response generally decreases and survival duration of the isolated retina is generally shortened. No significant difference between the effects of the three tested products has been found. These results are discussed in relation to, on the one hand, the mechanism of intraretinal generation of its electrophysiological signal and, on the other hand, the leukotrienes' role in inflammatory diseases of the retina. PMID- 2993186 TI - Superoxide generation by human blood leucocytes under the effect of cytolytic agents. AB - Human blood leucocytes generate large amounts of superoxide following stimulation by polyarginine, polyanetholesulphonate and mixtures of a variety of soluble agents. Generation of O2-. by the various "cocktails" of soluble ligands is markedly enhanced by cytochalasins A, B, C, D, E and F. The efficiency of cytochalasin A is, however, at least 50-fold greater than that of the other cytochalasins. Leucocytes that have been treated for a few minutes with the cytolytic agents saponin, digitonin and lysolecithin undergo lysis and lose their superoxide-producing capacities, when a variety of soluble ligands are employed to stimulate superoxide production. A partial reactivation of the superoxide producing capacities of the leucocytes can be achieved by adding NADPH. However, as the concentration of the cytolytic agents increases, reactivation of the cytochrome C reduction is less inhibitable by SOD, suggesting that cell lysis releases reductases of cytochrome C not connected with the superoxide-producing system of the leucocytes. Both saponin and digitonin can totally replace polyarginine as ingredients of the "cocktail," suggesting that these agents may also function as "priming agents" for superoxide production which can, however, further be enhanced by the addition of mixtures of soluble agents. Thus, leucocytes which had been lysed by membrane-active agents can nevertheless produce superoxide if adequate amounts of NADPH are added. PMID- 2993187 TI - Effect of growth hormone on the activity of some lysosomal enzymes in neutrophilic polymorphonuclear leukocytes of hypopituitary dwarfs. AB - Elevated activities of beta-D-glucuronidase, myeloperoxidase, and lysozyme were found in polymorphonuclear leukocytes (PMNs) of both hypopituitary dwarfs and normal subjects after the administration of growth hormone (GH), as compared to the activities in PMNs from blood drawn immediately before the administration of GH. During in vitro incubation, GH was able to inhibit the release of lysosomal enzymes from resting PMNs. This inhibition may be one of the reasons for the elevated lysosomal enzyme activities observed in PMNs after the administration of GH. GH can also affect hexose monophosphate shunt (HMPS) activity and superoxide production by PMNs. The activity of HMPS is stimulated by GH in resting PMNs, while in PMNs incubated with zymosan the GH inhibits both HMPS and superoxide production. PMID- 2993189 TI - [Atypical pneumonias--mycoplasmas, rickettsias, chlamydias, viruses]. PMID- 2993188 TI - Evaluation of a skin test for chicken pox. AB - Results with a VZV skin test as a marker of past infection were compared with histories of chicken pox and specific antibody detected by ELISA in 100 individuals--25 of whom were pediatric patients with malignant diseases. A negative or uncertain history was not reliable, neither were the skin test results among the oncology patients. However, among the normal individuals, the skin test when compared with the ELISA had a sensitivity of 85%, a specificity of 100%, and a positive predictive value of 100%. PMID- 2993191 TI - Correlation of corneal endothelial pump site density, barrier function, and morphology in wound repair. AB - After transcorneal freezing, physiologic function (pump and barrier) of the regenerating rabbit corneal endothelium was evaluated and compared with the morphologic differentiation that occurs during wound healing. Endothelial pump function was investigated utilizing the specific binding of tritiated ouabain to endothelial Na+/K+ ATPase (pump sites); the permeabilities of isolated de epithelialized corneas to labeled inulin and dextran were measured to determine endothelial barrier function. Endothelial recovery after transcorneal freezing can be described as a three-stage process. Stage one is characterized by the establishment of an initial coverage of the wound by pleomorphic spindle-shaped cells which form a functional but incomplete barrier and minimal pump site density. In stage two, the cells assume a flattened configuration consisting of irregular polygons and establish nearly normal pump capacity. In stage three, a significant remodeling of the monolayer continues despite the layer's early physiologic recovery. PMID- 2993190 TI - [Physiology and pathophysiology of lipoprotein metabolism--relation to hyperlipemias and arteriosclerosis]. PMID- 2993192 TI - Iontophoresis of epinephrine isomers to rabbit eyes induced HSV-1 ocular shedding. AB - Iontophoresis of 0.01% levo(-) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days induces ocular shedding of herpes simplex virus type 1 (HSV-1) in latently infected rabbits. In the present experiment, we tested dextro(+) and levo(-) epinephrine for their comparative effects on induced HSV-1 ocular shedding. One hundred percent of the eyes shed virus after either 0.01% dextro(+) or levo(-) epinephrine iontophoresis (8 min, 0.8 mAmp). However, the shedding frequency caused by 0.005% levo(-) epinephrine was significantly higher (P less than 0.05) than that by 0.005% dextro(+) epinephrine when the iontophoresis was conducted at 0.4 mAmp for 4 min. Iontophoresis of 0.001% levo(-) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days and iontophoresis of 0.001% dextro(+) epinephrine for 8 min at 0.8 mAmp once daily for 3 consecutive days did not induce HSV-1 ocular shedding in latently infected rabbits. The data suggest that the mechanism of induction of HSV-1 ocular shedding by epinephrine is correlated to the receptor potency of levo(-) epinephrine. PMID- 2993193 TI - Effects of cholinergic drugs and 4-aminopyridine on cat ciliary muscle contractility. AB - The effects of some cholinergic agents and 4-aminopyridine (4-AP) on neurally mediated contractions of in vitro cat ciliary muscle preparations were studied. The contractile response to trains of stimuli was enhanced by eserine and completely blocked by tetrodotoxin or atropine. Low concentrations of carbachol did not modify muscle resting tension but clearly attenuated contractile response to electrical stimuli, while higher concentrations increased the resting tonus leading to contracture which did not respond to further stimulation. 4-AP is known to be a potassium-channel blocking drug that increases neurotransmitter release at nerve terminals during the action potential. This substance exhibited a dose-related potentiation of the evoked ciliary muscle contractions without changing resting tension. The eventual reducing effect of 4-AP on the accommodative convergence/accommodation ratio (AC/A) is discussed in relation to its potential clinical application in certain strabismus patients. PMID- 2993194 TI - Comparison between a biochemical and a histochemical method for the detection of oestrogen receptor in breast carcinoma. PMID- 2993195 TI - Lateral differences in the GABAergic system of the rat striatum. AB - Asymmetric differences have been found in the pre- and postsynaptic activity of the GABAergic system of the left and right striata of the rat. 3H-GABA binding shows a higher dissociation constant (KD) and a higher number of sites (Bmax) in the left striatum than in the right. Moreover, 3H-diazepam binding seems to be more extensively activated by GABA in the right striatum suggesting a more sensitive postsynaptic GABAergic activity than on the left side. However, when the presynaptic marker (GAD activity) was measured, the asymmetry was in the opposite direction. The results provide further neurochemical evidence of the functional asymmetry of the rat brain. PMID- 2993196 TI - Primary intracranial tumors. Survey of incidence in the province of Trento in the years 1977-1981. AB - The incidence rate of primary intracranial tumors in the Province of Trento from 1977 to 1981 was 7.4 per 100,000 inhabitants per year. The age curve is as reported by other authors, with two peaks, one early and the other, much higher, at 50-59 years. This curve reflects that of malignant tumors in males. The age adjusted crude rate in males proved to be 7.4 (4.9 for malignant and 2.5 for benign tumors). The corresponding value in females was 7.4 (3.5 and 4.0). The most frequent tumors in males were astrocytomas, grades III and IV (M/F ratio = 1.6), and in females meningiomas (M/F ratio = 0.6). Hypophyseal adenomas, which accounted for 15.4% of all tumors, had an M/F ratio of 0.4. The factors likely to be responsible for the differences in incidence rates reported in the various surveys are discussed. PMID- 2993197 TI - Biometric estimation of chest wall thickness of females. AB - Optimal use of whole-body counting data to estimate pulmonary deposition of many of the actinides is dependent upon accurate measurement of the thickness of the chest wall because of severe attenuation of low-energy x rays and photons associated with the decay of these radionuclides. An algorithm for estimation of female chest wall thicknesses, verified by real-time ultrasonic measurements, has been derived based on the correlation of measured chest wall thickness and other common biometric quantities. Use of this algorithm will reduce the error generally associated with estimation of internal actinide deposition previously resulting from assuming an average chest wall thickness for all female subjects. PMID- 2993198 TI - Dependence of electrostatic diffusion of Rn progeny on environmental parameters. AB - In previous work it was established that electrostatic diffusion of Rn progeny attached to aerosols can be used for passive dosimetry by replacing the pump filter assembly in an active dosimeter by an electret. In the present work, results are presented on the dependence of activity collected by an electret on aerosol concentration, aerosol diameter, attachment rate of Rn progeny to aerosols, unattached fraction of Rn progeny, and Rn progeny concentrations. The electrostatic flow rate of Rn progeny is defined and all relationships analyzed in terms of this flow rate. The electrostatic flow rate is found to be reasonably constant for 0.03 micron less than particle diameter less than 0.09 micron, unattached fraction of 218Po less than 0.40 and 70,000/cm3 less than aerosol concentration less than 180,000/cm3; experimentally obtained data outside these limits are too few to draw any definite conclusions. The results are useful from a practical point of view because typical U mine conditions are described by these limits. PMID- 2993199 TI - Simultaneous measurement of Rn and its progeny in cave air by liquid scintillation techniques and alpha-ray spectrometry. PMID- 2993200 TI - Effect of electric fields on 220Rn progeny concentration. PMID- 2993201 TI - Surgical treatment for insulinoma: a study of 6 cases. PMID- 2993202 TI - Plasminogen activator: morphological evidence of binding, internalization and delivery to lysosomes in 3T3 mouse fibroblasts. AB - Using a direct conjugate of urokinase and ferritin, the binding has been followed at the plasma membrane and the internalization of urokinase into BALB/C-3T3 fibroblasts, cultured in plasminogen-free conditions. At 0 degree C, the conjugate was observed bound on both coated and uncoated cell surface regions as singlets, and small and large clusters. No binding was observed in the presence of excess native urokinase. The binding was impaired by preincubation of the conjugate with a competitive inhibitor of the catalytic site, suggesting an interaction between the receptor and the catalytic site of the enzyme. Within 1 min at 37 degrees C, urokinase clustered on coated regions of the plasma membrane. At 5 min after warming, ferritin was found on deeply indented coated pits and in both coated and uncoated vesicles close to the cell surface. By 10 min at 37 degrees C, ferritin particles were present in uncoated endosomes and in multivesicular bodies in the Golgi area. Within 10 min, the receptors on the surface strongly decreased. New receptors were observed on the membrane after 20 min at 37 degrees C. At this time, ferritin was observed both in endosomes or multivesicular bodies and in vesicles close to the plasma membrane. PMID- 2993203 TI - Localization of phosphatases in the rat parotid gland. PMID- 2993204 TI - Ultrahistochemical localization of adenylate cyclase activity in the electric organ of Torpedo marmorata. AB - The lead pyrophosphate precipitation technique was used to visualize adenylate cyclase activity with the electron microscope in unfixed electric organ and synaptosomes of Torpedo marmorata, with special attention to presynaptic membranes. Specificity of the deposition of reaction product was ensured by using 5'-adenylyl imidodiphosphate as substrate and 5'-guanylyl imidodiphosphate and sodium fluoride as activators. Under suitable conditions a reaction product was deposited on the Schwann cell, on presynaptic vesicles, on the inner side of membranes of cisternae and on glycogen granules of the presynaptic region of the endplate. In some cases, a precipitate was also found on postsynaptic membranes of the synaptic cleft and on mitochondria. In isolated synaptosomes localization of the reaction product was identical with that of minced tissue. However, most strikingly, on presynaptic membranes no precipitate was ever found, neither in pieces of electric organ nor in isolated synaptosomes. Furthermore, the extended membrane system of the postsynaptic region of the electroplax remained always free of lead pyrophosphate precipitate. PMID- 2993206 TI - Immunocytochemical study on the cytoplasmic side of cell membranes infected with vesicular stomatitis virus by quick-freezing and deep-etching replica method. AB - Synthesized N protein of vesicular stomatitis virus (VSV) is associated with replicated viral genomes in the infected cells. The cytoplasmic side of cell membranes was examined by quick-freezing and deep-etching replica method, in order to clarify the localization of VSV genomes. Control or infected monolayer Vero cells were fixed in 2% paraformaldehyde, scraped and centrifuged to make pellets. A drop of the cell pellet was put between two glass coverslips, which were coated with 3-aminopropyl triethoxy silane and glutaraldehyde. The cells were consequently split open and postfixed in the mixture of glutaraldehyde and paraformaldehyde. Some inside-out cell membranes on the coverslips were immunostained with anti-N monoclonal antibody directly coupled to gold particles. Others were immunostained with anti-N monoclonal antibody and rabbit anti-mouse IgG coupled to peroxidase and fixed again in glutaraldehyde. They were incubated in diaminobenzidine and hydrogen peroxide solution for 1 min. All of them were infiltrated with 10% methanol in distilled water and quickly frozen in a mixture of isopentane and propane cooled by liquid nitrogen. Such preparations were deep etched and shadowed by platinum and carbon. Although many cell organelles were found to be associated with the cytoplasmic side of cell membranes in the normal Vero cells, few cell organelles were attached to it in the infected cells. On the contrary, special strand structures were identified, which could be immunostained with anti-N monoclonal antibody. It is concluded that platinum replicas have sufficient resolution to identify the VSV genomes coated with N protein and that these nucleocapsids can be associated with the cytoplasmic side of cell membranes in the infected cells. PMID- 2993205 TI - Sex different cytochrome-c uptake in the proximal tubule of the rat kidney. AB - The present study deals with the dose- and time-dependent uptake of cytochrome c (CYT c) in the proximal tubule of the rat kidney, and shows that there are segment and sex differences in the reabsorption of CYT c. Rats of both sexes were intravenously injected with different doses of CYT c (0.75-9.0 mg per 100 g body weight), and the kidneys were investigated by light and electron microscopy at different times (3 min, 10 min, and 2 h) after the injection. After 3 and 10 min, CYT c was demonstrated in apical vacuoles of different sizes and in some lysosomes of the S1 and S2 segments, whereas after 2 h, CYT c was found only in lysosomes of all three segments of the proximal tubule. At these times, the S1 segment contained more CYT c than the S2 and S3 segments. However, 2 h after the injection of 6 or 9 mg CYT c, the differences between the S1 and S2 segments disappeared almost completely, due to a strong lysosomal accumulation of CYT c in the S2 segment. At all studied times and CYT-c doses, the S3 segment contained less CYT c than the S1 and S2 segments. On the whole, different levels of CYT-c reabsorption were found in the different segments of the proximal tubule, which was saturable with increasing CYT-c doses, i.e. firstly in the proximal and then in the distal parts of the proximal tubule. Two hours after the injection of CYT c, a difference between males and females was observed, with the lysosomes of the S1 and S2 segments of females containing more CYT c than those of males. Thus, more CYT c was reabsorbed in the proximal tubule of females than in that of males. PMID- 2993207 TI - Increased frequency of IgG heavy chain marker Glm(2) and of HLA-B8 in Lambert Eaton myasthenic syndrome with and without associated lung carcinoma. AB - In view of the evidence for an autoimmune pathogenesis of the Lambert-Eaton myasthenic syndrome, we have sought associations with IgG heavy chain allotypes (Gm) and HLA antigens in 30 patients, of whom 20 had evidence of lung carcinoma (histologically proven small ("oat") cell type in 17). A highly significant overall increase in frequency of Glm(2) (chi 2 = 10.95; p less than 0.001; n = 30) and of HLA-B8 (chi 2 = 19.07; p less than 0.001; n = 23) was observed. These two factors apparently occurred independently of each other. The Glm(2) frequency in 36 non-myasthenic small cell carcinoma cases was the same as in a control panel (n = 167). We conclude that Glm(2) and HLA-B8 both associate with increased susceptibility to the Lambert-Eaton myasthenic syndrome, and suggest that Glm(2) may be in linkage disequilibrium with a limited number of VH genes coding for antibodies to restricted antigenic determinants at the nerve terminals, which may be shared by the carcinoma cells. PMID- 2993208 TI - Motor control in humans with large-fiber sensory neuropathy. AB - Upper limb motor control was evaluated in a series of patients with a large-fiber sensory neuropathy associated with impaired position, vibration and cutaneous sensation and absence of deep tendon reflexes. Muscular strength was normal or only minimally affected. In studies of wrist movement it was found that both postural maintenance and accuracy of wrist displacement were heavily dependent on visual guidance. Without vision the limb would drift in almost random directions, and during intended movements both the trajectory and movement end-point were abnormal. The defects in posture and voluntary movement control were reflected in the inability of patients to maintain consistent levels or emit consistent patterns of muscle activity. It is concluded that whereas central motor commands are sufficient to initiate movements proprioceptive afferent inputs are important for accurate postural maintenance and the fine control of movement. PMID- 2993209 TI - Quantitative dose-response analysis of salivary function following radiotherapy using sequential RI-sialography. AB - A total of 216 99mTc-pertechnetate-sialographies, by applying stimulations by citric acid, were performed for studying radiation effects on the salivary glands in 145 patients. From a scintigram of the salivary gland, the time-activity curves were obtained for analysis of salivary function. The shape of the time activity-curve with respect to stimulus-response pattern was classified into four types according to the degree of radiation-induced dysfunction. We found that the acid stimulation was enhancing accumulation of the isotope into the salivary gland as well as influencing salivary discharge. A ratio (Rc) of pre- and post stimulation counts permitted a construction of quantitative dose-response curves; the curve for the parotids had a slight rise of Rc's over 0-20 Gy, whereas that for the submandibular glands had a significant rise over 10-30 Gy; in both glands the rise of Rc's was followed by a steep fall. The parotids appeared to be more sensitive to radiation than the submandibular glands at 0-3 months following 20 70 Gy, but after 3 months both glands were similarly impaired. The gland volume irradiated was also of great importance in determining the degree of salivary dysfunction. The maximum uptakes averaged per unit area (salivary-to-background ratio) and determined during the initial 30 minutes were greater in the submandibular than in the parotid glands following greater than 20 Gy. In both glands the maximum uptakes were gradually impaired with time over 3-36 months following 50-70 Gy. PMID- 2993211 TI - Peripheral polyneuropathy in a dog with functional islet B-cell tumor and widespread metastasis. AB - A 12-year-old female Irish Setter was examined because of recurrent episodes of hindlimb weakness. The dog was not ataxic, but had generalized muscular atrophy, decreased patellar reflexes, and slow proprioception. Blood glucose content was low (32 mg/dl) and the amended insulin-glucose ratio was high (9,600). Electromyographic studies showed evidence of polyneuropathy. The diagnosis was functional islet B-cell tumor with peripheral neuropathy. Exploratory laparotomy was performed. Widespread metastases were observed and the dog was euthanatized because of the poor prognosis. The dog was necropsied and the diagnosis was confirmed on the basis of microscopic findings. PMID- 2993210 TI - Radioprotective activities of diethyldithiocarbamate. PMID- 2993212 TI - Clinical evaluation of sodium bicarbonate, sodium L-lactate, and sodium acetate for the treatment of acidosis in diarrheic calves. AB - Thirty-six dehydrated diarrheic neonatal calves were used to study the effects of various alkalinizing compounds on acid-base status, the changes in central venous pressure (CVP) in response to rapid IV infusion of large volumes of fluid, and the correlation of acid-base (base deficit) status, using a depression scoring system with physical determinants related to cardiovascular and neurologic function. Calves were allotted randomly to 4 groups (9 calves/group). Over a 4 hour period, each calf was given two 3.6-L volumes (the first 3.6 L given in the first hour) of a polyionic fluid alone (control group) or were given the polyionic fluid with sodium bicarbonate, sodium L-lactate, or sodium acetate added (50 mmol/L). Acid-base status, hematologic examination, and biochemical evaluations were made immediately before infusion of each fluid (at entry) and after 3.6, 4.8, and 7.2 L of fluid had been given. Compared with control values, bicarbonate, lactate, and acetate had significantly greater alkalinizing effects on pH (P less than 0.01) and base deficit (P less than 0.01) after 3.6, 4.8, and 7.2 L of fluid were given. Bicarbonate had the most rapid alkalinizing effect and induced greater changes in base deficit (P less than 0.01) than did acetate or lactate at each of the 3 administered fluid volumes evaluated. Acetate and lactate had similar alkalinizing effects on blood. Rehydration alone did not improve acid-base status. The CVP was elevated in 10 (28%) of the 36 calves after 1 hour of fluid (3.6 L) administration, but significant differences in body weight, PCV, and clinical condition or depression score at entry were not found between calves with elevated CVP and those with normal CVP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993213 TI - Pancreatic beta-cell carcinoma with renal metastasis in a dog. AB - Severe hypoglycemia associated with a metastatic pancreatic beta-cell carcinoma was found in an adult female American Eskimo dog. Metastatic tumor was found in the renal glomeruli as opposed to the more common hepatic metastatic site. PMID- 2993214 TI - c-myc Gene amplification in primary stomach cancer. AB - Fourteen human primary stomach cancer tissues were screened by Southern blot hybridization using six oncogene probes (myc, myb, H-ras, K-ras, abl, mos), and an amplification of c-myc oncogene was found in one tissue. This is the first report of c-onc amplification in primary stomach cancer tissue. PMID- 2993215 TI - Hepatitis B surface antigen in hepatocellular carcinoma. AB - Hepatitis B surface antigen (HBs-Ag) was detected histologically (by victoria blue-nuclear fast red staining) in the liver in 20-40% of liver cirrhosis and/or hepatocellular carcinoma (hepatoma) cases. In hepatoma cases, HBs-Ag was usually found in non-cancerous areas of the liver. However, some HBs-Ag positive cells were also found dispersed in cancerous areas; these were regarded as HBs-Ag infected non-cancerous hepatic cells remaining undestroyed. There were a few cases where inconsistency was found between the results of the immunofluorescence technique using anti HBs-Ag serum and victoria blue staining in detecting HBs-Ag. This phenomenon was rare in non-cancerous areas, but was relatively frequent in cancerous areas. PMID- 2993216 TI - Antitumor activity of Sphaerotilus natans and its slime fraction in mice. AB - Antitumor activity of the whole cell and slime of an aquatic sheathed bacterium, Sphaerotilus natans IAM 12068, against ascites form of Ehrlich carcinoma in ddY mice was investigated. Intraperitoneal injection of whole cells and the slime fraction showed remarkable antitumor activity against mice inoculated with 10(4) to 10(5) tumor cells, the slime fraction being more effective. To examine the chemical nature of the active principle in the slime fraction, separation by Sepharose 4B gel filtration was carried out and two fractions designated as GF-P 1 and GF-P-2, which are mainly composed of protein, carbohydrate, and lipid, were obtained. GF-P-1 fraction, which contains large amounts of fucose and unidentified sugar as neutral sugar, showed marked antitumor activity at half the dose of the slime fraction, whereas the antitumor activity of GF-P-2, which is composed mainly of protein, was weak. This finding indicates that GF-P-1 fraction of S. natans slime may be a main active principle. The consistently demonstrable antitumor activity of GF-P-1 was abrogated by treatment of mice with silica, an anti-macrophage agent, suggesting that the antitumor activity of GF-P-1 depends on the activation of macrophages. PMID- 2993217 TI - A comparative analysis of cAMP-dependent protein kinase regulatory subunits in sea urchin and rat spermatozoa. AB - 8-Azido cAMP photoaffinity labeling of cAMP-dependent protein kinase regulatory subunits (R1 = 49 K;R2 = 55K) was done on spermatozoa from species lacking, and species containing an epididymis. Spermatozoa from sea urchin and trout contained only R1, while rat caudaepididymal spermatozoa contained both R1 and R2 subunits. This was established by the Mr value of the 8-azido cAMP photolabeled moieties, and a biochemical analysis based on the known differences of protein-nucleotide interactions of Type I and II cAMP-dependent protein kinases. Sea urchin and trout sperm R1 subunits were similar to mammalian sperm R1 subunits in co migration on SDS-polyacrylamide gels and in both saturation and specificity of nucleotide binding. Calcium enhanced photoprobe binding to rat R1 and R2 subunits and to sea urchin R1 subunit without revealing a sea urchin R2 subunit. Likewise, phosphodiesterase incubation of sea urchin and trout spermatozoa prior to photolabeling did not reveal R2 subunits. These data suggest that the cAMP regulation of sperm physiology may require R1 subunit in species both with and without an epididymis. Further taxonomic study is necessary to determine whether evolutionary acquisition of the epididymis and internal fertilization may have created unique environments favoring the addition of sperm R2 regulatory subunits of cAMP-dependent protein kinase. PMID- 2993218 TI - Griseolic acid, an inhibitor of cyclic adenosine 3',5'-monophosphate phosphodiesterase. I. Taxonomy, isolation and characterization. AB - Griseolic acid, a potent inhibitor of cyclic adenosine 3',5'-monophosphate (cyclic AMP) phosphodiesterase (EC 3.1.4.17) was isolated from the cultured broth of Streptomyces griseoaurantiacus SANK 63479. Griseolic acid has an adenine moiety and two carboxyl groups and possesses the molecular formula C14H13N5O8. Griseolic acid inhibited cyclic AMP phosphodiesterase competitively with regard to cyclic AMP, the substrate and the resulting Ki value was 0.26 microM. Griseolic acid showed the most potent inhibitory effect on rat aorta cyclic AMP phosphodiesterase among the enzymes of several rat organs tested. The plasma level of cyclic AMP was increased when griseolic acid was subcutaneously injected rats. PMID- 2993219 TI - Griseolic acid, an inhibitor of cyclic adenosine 3',5'-monophosphate phosphodiesterase. II. The structure of griseolic acid. AB - Griseolic acid, a potent inhibitor of cyclic adenosine 3',5'-monophosphate phosphodiesterase, was isolated from the fermentation broth of Streptomyces griseoaurantiacus SANK 63479. Treatment of griseolic acid with HCl-MeOH gave adenine and pseudo-sugar. The structure of griseolic acid, adenine nucleoside type structure, was elucidated by chemical degradation and X-ray analysis, and was shown to be structure 1. PMID- 2993220 TI - Peripheral auditory adaptation and fatigue: a model oriented review. AB - A model is introduced for auditory adaptation based on the stochastic models that are widely documented for birth and death processes and used as a vehicle to review single unit and compound action potential studies as well as various models for adaptation and forward masking. It appears that such a model inherently incorporates a relation between the perstimulatory adaptation time constant, the poststimulatory recovery time constant and the ratio between adapted firing rate and onset firing rate. The knowledge of any two of these parameters allows the prediction of the third one. The model takes into account postsynaptic membrane properties. The model based on Markov assumptions is a linear one. Although onset firing rates depend on a nonlinear way upon stimulus level, the above mentioned time constants and the adapted rate to onset rate ratio are intensity independent. The idea is put forward and tested that a comparable situation exists for the depression effects produced by mild auditory fatigue. The apparent findings that time constants tend to be intensity dependent for auditory fatigue is explained on basis of an interaction effect of dependent exponential processes. PMID- 2993221 TI - Changes and interrelationships among luteal LH receptors, adenylate cyclase activity and phosphodiesterase activity during the bovine estrous cycle. AB - Changes and interrelationships among the cellular mechanisms that may regulate luteal progesterone synthesis during the bovine estrous cycle were studied. Corpora lutea (CL) were enucleated from 30 cows via a transvaginal incision on d 4, 7, 10, 13, 16 and 19 following estrus (estrus = d 0). Mean corpus luteum weight, and luteal progesterone and plasma progesterone concentrations increased (P less than .05) from d 4 to 7 or 10, and declined following luteal regression (d 19). Unoccupied LH receptor (UOR) concentrations increased (P less than .05) more than fourfold from d 4 (38 fmol/mg protein) to d 16 (173 fmol/mg protein) before declining (P less than .05) by d 19 (30 fmol/mg protein). The dissociation constant for UOR concentrations increased (P less than .05) during the estrous cycle. Concentrations of occupied LH receptors (OR) were less (P less than .05) on d 7 than other days; however, the total number of OR/CL increased fourfold from d 4 (47 fmol/CL) to d 10 (221 fmol/CL) and remained similar thereafter. Basal adenylate cyclase, LH-activated adenylate cyclase, and Gpp(NH)p-activated adenylate cyclase activities were greatest (P less than .05) on d 7 to 16 as compared with d 4 and 19. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity on d 7, 10, 13, and 16; whereas, Gpp(NH)p stimulated (P less than .05) adenylate cyclase activity relative to basal activity at each period. Phosphodiesterase was 46% greater on d 19 as compared with d 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993222 TI - Estimation of ventilation-perfusion inequality by inert gas elimination without arterial sampling. AB - Estimation of ventilation-perfusion (VA/Q) inequality by the multiple inert gas elimination technique requires knowledge of arterial, mixed venous, and mixed expired concentrations of six gases. Until now, arterial concentrations have been directly measured and mixed venous levels either measured or calculated by mass balance if cardiac output was known. Because potential applications of the method involve measurements over several days, we wished to determine whether inert gas levels in peripheral venous blood ever reached those in arterial blood, thus providing an essentially noninvasive approach to measuring VA/Q mismatch that could be frequently repeated. In 10 outpatients with chronic obstructive pulmonary disease, we compared radial artery (Pa) and peripheral vein (Pven) levels of the six gases over a 90-min period of infusion of the gases into a contralateral forearm vein. We found Pven reached 90% of Pa by approximately 50 min and 95% of Pa by 90 min. More importantly, the coefficient of variation at 50 min was approximately 10% and at 90 min 5%, demonstrating acceptable intersubject agreement by 90 min. Since cardiac output is not available without arterial access, we also examined the consequences of assuming values for this variable in calculating mixed venous levels. We conclude that VA/Q features of considerable clinical interest can be reliably identified by this essentially noninvasive approach under resting conditions stable over a period of 1.5 h. PMID- 2993223 TI - Pure gelatin microcarriers: synthesis and use in cell attachment and growth of fibroblast and endothelial cells. AB - A new type of microcarrier was described using bead emulsion-polymerization techniques. An aqueous solution of gelatin and glutaraldehyde was dispersed in a hydrophobic phase of mineral oil, using Triton X-114 as an emulsifier, and polymerization was initiated. The resultant spherical beads, composed entirely of gelatin, showed excellent mechanical stability to ethanol drying, sterilization, and long-term use in microcarrier spinner cultures. The solid gelatin microcarriers supported the growth of L-929 fibroblast, swine aorta endothelial, human umbilical endothelial, and HeLa-S3 cultures with no adverse effects on cell morphology or growth. The beads were transparent in growth medium and attached cells were clearly visualized without staining. The beads were also compatible with techniques for scanning electron microscopy. Collagenase could be used to entirely digest the gelatin beads, leaving the cells free from microcarriers and suspended in solution while retaining 98% cell viability. The results further showed that after collagenase treatment the cells would populate fresh gelatin microcarriers and grow to confluence. Cell attachment kinetics revealed that the endothelial cells attached to the gelatin beads at the same rate as to tissue culture plates, whereas the fibroblast cells attached to the beads more slowly. However, once the fibroblast cells were attached to the gelatin microcarriers they spread and grew normally. PMID- 2993224 TI - Linkage of genes for laminin B1 and B2 subunits on chromosome 1 in mouse. AB - We have used cDNA clones for the B1 and B2 subunits of laminin to find restriction fragment length DNA polymorphisms for the genes encoding these polypeptides in the mouse. Three alleles were found for LamB2 and two for LamB1 among the inbred mouse strains. The segregation of these polymorphisms among recombinant inbred strains showed that these genes are tightly linked in the central region of mouse Chromosome 1 between Sas-1 and Ly-m22, 7.4 +/- 3.2 cM distal to the Pep-3 locus. There is no evidence in the mouse for pseudogenes for these proteins. PMID- 2993225 TI - Guanine nucleotide--an intracellular traffic cop. PMID- 2993226 TI - Determination of total dietary fiber in foods and food products: collaborative study. AB - A collaborative study was conducted to determine the total dietary fiber (TDF) content of food and food products, using a combination of enzymatic and gravimetric procedures. The method was basically the same as published earlier (J. Assoc. Off. Anal. Chem. (1984) 67, 1044-1052), with changes in the concentration of alcohol and buffers, time of incubation, sample preparation, and some explanatory notes, all with the intent of decreasing the coefficient of variation (CV) of the method. Duplicate blind samples of soy isolate, white wheat flour, rye bread, potatoes, rice, wheat bran, oats, corn bran, and whole wheat flour were analyzed by 9 collaborators. TDF was calculated as the weight of the residue minus the weight of protein and ash. CV values of the data from all laboratories for 7 of the samples ranged from 1.56 to 9.80%. The rice and soy isolate samples had CV values of 53.71% and 66.25%, respectively; however, each sample contained only about 1% TDF. The enzymatic-gravimetric method for determining TDF has been adopted official first action. PMID- 2993227 TI - Identification and characterization of the mutL and mutS gene products of Salmonella typhimurium LT2. AB - The gene products of the mutL and mutS loci play essential roles in the dam directed mismatch repair in both Salmonella typhimurium LT2 and Escherichia coli K-12. Mutations in these genes result in a spontaneous mutator phenotype. We have cloned the mutL and mutS genes from S. typhimurium by generating mutL- and mutS specific probes from an S. typhimurium mutL::Tn10 and an mutS::Tn10 strain and using these to screen an S. typhimurium library. Both the mutL and mutS genes from S. typhimurium were able to complement E. coli mutL and mutS strains, respectively. By a combination of Tn1000 insertion mutagenesis and the maxicell technique, the products of the mutL and mutS genes were shown to have molecular weights of 70,000 and 98,000 respectively. A phi (mutL'-lacZ+) gene fusion was constructed; no change in the expression of the fusion could be detected by treatment with DNA-damaging agents. In crude extracts, the MutS protein binds single-stranded DNA, but not double-stranded DNA, with high affinity. PMID- 2993228 TI - Plasmid genes required for microcin B17 production. AB - The production of the antibiotic substance microcin B17 (Mcc) is determined by a 3.5-kilobase DNA fragment from plasmid pMccB17. Several Mcc- mutations on plasmid pMccB17 were obtained by both transposon insertion and nitrosoguanidine mutagenesis. Plasmids carrying these mutations were tested for their ability to complement Mcc- insertion or deletion mutations on pMM102 (pMM102 is a pBR322 derivative carrying the region encoding microcin B17). Results from these experiments indicate that at least four plasmid genes are required for microcin production. PMID- 2993229 TI - Physical characterization of the cloned protease III gene from Escherichia coli K 12. AB - Analysis of the cloned protease III gene (ptr) from Escherichia coli K-12 has demonstrated that in addition to the previously characterized 110,000-Mr protease III protein, a second 50,000-Mr polypeptide (p50) is derived from the amino terminal end of the coding sequence. The p50 polypeptide is found predominantly in the periplasmic space along with protease III, but does not proteolytically degrade insulin, a substrate for protease III. p50 does not appear to originate from autolysis of the larger protein. Protease III is not essential for normal cell growth since deletion of the structural gene causes no observed alterations in the phenotypic properties of the bacteria. A 30-fold overproduction of protease III does not affect cell viability. A simple new purification method for protease III is described. PMID- 2993230 TI - Genetic recombination of bacterial plasmid DNA: effect of RecF pathway mutations on plasmid recombination in Escherichia coli. AB - Tn5 insertion mutations in the recN gene, and in what appears to be a new RecF pathway gene designated recO and mapping at approximately 55.4 min on the standard genetic map, were isolated by screening Tn5 insertion mutations that cotransduced with tyrA. The recO1504::Tn5 mutation decreased the frequency of recombination during Hfr-mediated crosses and increased the susceptibility to killing by UV irradiation and mitomycin C when present in a recB recC sbcB background, but only increased the sensitivity to killing by UV irradiation when present in an otherwise Rec+ background. The effects of these and other RecF pathway mutations on plasmid recombination were tested. Mutations in the recJ, recO, and ssb genes, when present in otherwise Rec+ E. coli strains, decreased the frequency of plasmid recombination, whereas the lexA3, recAo281, recN, and ruv mutations had no effect on plasmid recombination. Tn5 insertion mutations in the lexA gene increased the frequency of plasmid recombination. These data indicate that plasmid recombination events in wild-type Escherichia coli strains are catalyzed by a recombination pathway that is related to the RecF recombination pathway and that some component of this pathway besides the recA gene product is regulated by the lexA gene product. PMID- 2993231 TI - F factor inhibition of conjugal transfer of broad-host-range plasmid RP4: requirement for the protein product of pif operon regulatory gene pifC. AB - By the use of deletions, point mutations, and gene fusions, we show that the protein product of the F factor pifC gene is responsible for F factor inhibition of plasmid RP4 conjugal transfer. Deletion analysis of pif sequences carried by pSC101-F chimeric plasmids demonstrated that removal of all or part of the pifC coding sequence greatly decreased or abolished the ability of these plasmids to inhibit RP4 transfer. Amber mutations in the pifC gene eliminated inhibition in an Su- host strain but not in and Su+ (supF) host. Plasmids carrying nonpolar pifC mutations did not decrease the efficiency of RP4 transfer when present in trans. Whereas pifC+ plasmids inhibited RP4 transfer, the presence of RP4 in the same cell as F' lac increased F'lac Pif activity approximately 1,000-fold. This effect most likely resulted from the binding of the pifC product to RP4 DNA and concomitant derepression of the F factor pif operon. PifC inhibited trans mobilization of pMS204, a nonconjugative plasmid carrying the RP4 oriT locus, by the RP1 derivative pUB307. pMS204 had no trans effect on pif operon expression, whereas pUB307 increased F'lac Pif expression, as did RP4. Our results suggest that the pifC product inhibits expression of one or more RP4 genes, the products of which are required for conjugal transfer of RP4 and are required in trans for mobilization of nonconjugal RP4 oriT containing plasmids. PMID- 2993232 TI - Isolation and expression of the Escherichia coli gene encoding malate dehydrogenase. AB - An oligodeoxynucleotide specific for a pentapeptide sequence corresponding to amino acid residues 32 through 36 of Escherichia coli malate dehydrogenase was chemically synthesized and used to identify the mdh gene on plasmid pLC32-38 from the Clarke-Carbon recombinant library. Cells transformed with this plasmid exhibited a 10-fold increase in malate dehydrogenase activity. A 1.2-kilobase PvuII fragment which hybridized with the oligodeoxynucleotide probe was subcloned, and the identity of the mdh structural gene was confirmed by partial nucleotide sequence analysis. The expression of the mdh gene, as measured by both Northern blotting and enzyme assays, was found to vary over a 20-fold range with different culture conditions. PMID- 2993233 TI - Identification of the Escherichia coli deoR and cytR gene products. AB - The protein products encoded by the Escherichia coli deoR and cytR structural genes have been identified based on results obtained from E. coli maxicells harboring (i) recombinant plasmids carrying wild-type deoR and cytR genes, (ii) deletion derivatives of the deoR+ and cytR+ plasmids, (iii) plasmids containing site-specific mutations in the deoR and cytR structural genes, and (iv) plasmids which have transposon Tn1000 inserted into the deoR and cytR structural genes. Analysis of the protein profiles obtained from all the maxicell experiments demonstrated that the deoR gene encodes a protein with a subunit molecular weight of 30,500 and that the product of the cytR gene is a protein with a subunit molecular weight of 37,000. PMID- 2993234 TI - Thermoinducible transcription system for Bacillus subtilis that utilizes control elements from temperate phage phi 105. AB - We describe a thermoinducible-expression system for Bacillus subtilis which utilized an early promoter-operator sequence from temperate phage phi 105 and the thermolabile prophage repressor from the phage variant phi 105 cts23. The system operated at the transcriptional level to control expression in B. subtilis of the cat-86 gene derived from Bacillus pumilis. Details of the strategies used to isolate the early phage promoter are described. This promoter lay in close proximity to the prophage repressor gene on the phi 105 genome. The sequence of the early promoter differed from that of the vegetative B. subtilis consensus promoter by 1 base pair in both the -10 and -35 regions. We also present evidence that our phage-derived expression system could function in Escherichia coli to effect thermoinducible expression of the galK gene. PMID- 2993235 TI - Identification of the merR gene of R100 by using mer-lac gene and operon fusions. AB - Transcriptional (operon) and translational (gene) fusions between the R100 merR gene and lacZ were constructed in vitro in a pBR322 plasmid carrying the mer genes derived from plasmid R100. The translational fusions were oriented in the opposite direction to and divergently from the merTCAD genes. This shows that the reading frame previously thought to be merR was incorrect. Expression of the gene fusion was repressed in trans by a compatible plasmid carrying the R100 merR+ gene, as was a similarly oriented transcriptional fusion. In contrast, expression of beta-galactosidase by the lac fragment located at the same site but in the opposite orientation was at a lower level and was not repressed by merR+. PMID- 2993236 TI - Regulation of single and multicopy his operons in Escherichia coli. AB - We fused segments of the Escherichia coli his regulatory region to galK in single copy and multicopy vectors. These fusions demonstrated that (i) derepression of his by histidine starvation is due exclusively to attenuation; (ii) the his promoter is metabolically regulated; and (iii) both regulatory systems operate when the his regulatory region is present on a multicopy plasmid. Thus, there is no evidence for titration of his regulatory elements. Deletions of the his anti attenuator region, carried on multicopy plasmids, cause low-level galK expression. This expression is not stimulated by histidine starvation, but is growth rate dependent. We replaced the his attenuator with the efficient lambda terminator, to. In the context of the his regulatory region, however, lambda to only partially terminates transcription. PMID- 2993237 TI - Functional identification of the fatty acid reductase components encoded in the luminescence operon of Vibrio fischeri. AB - A clone of DNA, obtained from the luminescent bacterium Vibrio fischeri ATCC 7744 and inserted into pBR322, was found to express luminescence in Escherichia coli. Polypeptides involved in biosynthesis of the fatty aldehyde substrate for the light reaction were identified by fatty acid acylation of proteins synthesized in E. coli from the recombinant plasmid. The cloned region was similar to that reported for the V. fischeri MJ1 luminescence system (Engebrecht et al., Cell 32:773-781), except for some differences in endonuclease restriction sites and the requirement of a lower temperature for the expression of light in our cloned system. Fatty acid reductase activity could be detected in extracts of E. coli harboring the recombinant plasmid but not in extracts of the parental V. fischeri strain. Using in vivo labeling with [3H]tetradecanoic acid, we showed that the acylated polypeptides synthesized in the cloned system corresponded to the labeled polypeptides in V. fischeri (34, 42, and 54 kilodaltons) and that they could only be detected after induction of luminescence. These results provide direct evidence that the genes coding for the fatty acid reductase polypeptides are an integral part of the luminescence operon in the V. fischeri luminescence system. PMID- 2993238 TI - Identification of livG, a membrane-associated component of the branched-chain amino acid transport in Escherichia coli. AB - Branched-chain amino acids are transported into Escherichia coli by two osmotic shock-sensitive systems (leucine-isoleucine-valine and leucine-specific transport systems). These high-affinity systems consist of separate periplasmic binding protein components and at least three common membrane-bound components. In this study, one of the membrane-bound components, livG, was identified. A toxic analog of leucine, azaleucine, was used to isolate a large number of azaleucine resistant mutants which were defective in branched-chain amino acid transport. Genetic complementation studies established that two classes of transport mutants with similar phenotypes, livH and livG, were obtained which were defective in one of the membrane-associated transport components. Since the previously cloned plasmid, pOX1, genetically complemented both livH and livG mutants, we were able to verify the physical location of the livG gene on this plasmid. Recombinant plasmids which carried different portions of the pOX1 plasmid were constructed and subjected to complementation analysis. These results established that livG was located downstream from livH with about 1 kilobase of DNA in between. The expression of these plasmids was studied in minicells; these studies indicate that livG appears to be membrane bound and to have a molecular weight of 22,000. These results establish that livG is a membrane-associated component of the branched-chain amino acid transport system in E. coli. PMID- 2993239 TI - Streptococcal phosphoenolpyruvate: sugar phosphotransferase system: purification and characterization of a phosphoprotein phosphatase which hydrolyzes the phosphoryl bond in seryl-phosphorylated histidine-containing protein. AB - Histidine-containing protein (HPr) of gram-positive bacteria was found to be phosphorylated at a seryl residue (P-ser-HPr) in an ATP-dependent reaction catalyzed by a protein kinase (J. Deutscher and M. H. Saier, Jr., Proc. Natl. Acad. Sci. U.S.A. 80:6790-6794, 1983). Here we describe the purification and characterization of a soluble enzyme of Streptococcus faecalis which splits the phosphoryl bond in P-ser-HPr. The enzyme has a molecular weight of ca. 7.5 X 10(4), as determined by its migration behavior on a Sephacryl S-200 column. On native polyacrylamide gels the purified enzyme produced only one protein band. On sodium dodecyl sulfate-polyacrylamide gels we found one major protein band of molecular weight 2.9 X 10(4) and two minor protein bands of molecular weights 2.3 X 10(4) and 7 X 10(4). Fructose 1,6-diphosphate, which stimulated the ATP dependent, protein kinase-catalyzed phosphorylation of HPr, had no effect on the phosphatase activity. Other glycolytic intermediates also had no effect. However, inorganic phosphate, which inhibited the ATP-dependent HPr kinase, stimulated the P-ser-HPr phosphatase. EDTA at a concentration of 0.1 mM completely inhibited the phosphatase. Divalent cations like Mg2+, Mn2+, and Co2+ overcame the inhibition by EDTA. Fe2+, Zn2+, and Cu2+ had no effect, whereas Ca2+ slightly inhibited the phosphatase. ATP was also found to inhibit the phosphatase. Under conditions in which ATP severely inhibited the phosphatase, ADP was found to have no effect on the enzyme activity. Besides P-ser-HPr of S. faecalis, the phosphatase was also able to hydrolyze the phosphoryl bond in P-ser-HPr of Streptococcus lactis, Staphylococcus aureus, Bacillus subtilis, Streptococcus pyogenes, and Lactobacillus casei. Phosphoenolpyruvate-dependent o-nitrophenyl-beta-D galactopyranoside phosphorylation, catalyzed by the S. aureus phosphoenolpyruvate:lactose phosphotransferase system, was about 150-fold decreased in the presence of P-ser-HPr of S. aureus, as compared with HPr. However, when P-ser-HPr was first incubated with P-ser-HPr phosphatase to allow complete hydrolysis of the phosphoryl bond, it had the same activity as HPr. Besides this cytoplasmic phosphoprotein phosphatase, we detected a membrane-bound phosphatase which also hydrolyzed the phosphoryl bond in P-ser-HPr. PMID- 2993241 TI - Suicide vector for transposon mutagenesis in Pseudomonas solanacearum. AB - A suicide vector was constructed by cloning the transfer genes of the wide-host range (IncW group) plasmid R388 into the BamHI site of pBR325. This plasmid can deliver Tn5 into Pseudomonas solanacearum at frequencies ranging from 10(-6) to 10(-9) per recipient. PMID- 2993240 TI - Molecular cloning and structure of the gene for 7 beta-(4 carboxybutanamido)cephalosporanic acid acylase from a Pseudomonas strain. AB - A Pseudomonas strain produced an enzyme capable of deacylating 7 beta-(4 carboxybutanamido)cephalosporanic acid to 7-aminocephalosporanic acid in response to glutaric acid. The gene for the enzyme was cloned within the PstI site of pBR325 as a 7.35-kilobase-pair DNA segment from a mutant of this strain whose enzyme is produced constitutively. The gene expression in the primary clone appeared to be low in Escherichia coli but was significantly enhanced by reducing the size of the initial segment coupled with E. coli promoters. Subsequent subcloning resulted in localization of the gene to a 2.45-kilobase-pair fragment. Three clone-specific polypeptides with molecular weights of ca. 16,000, 54,000, and 70,000 were shown by maxicell analysis. The former two corresponded to the small and large subunits of the purified enzyme from the Pseudomonas strain, and the third polypeptide was suggested to be their precursor. This was supported by DNA sequence study together with amino acid sequencing of the amino terminus of both subunits: the sequences for the small and large subunits were localized contiguously in this order on the structural gene without termination codons between them. The nucleotide sequence also disclosed the presence of a signallike sequence preceding that for the small subunit, consistent with the previous observation that the enzyme might be periplasmic in the Pseudomonas strain. Those results suggest a process for the formation of an active enzyme complex from a precursor through two steps of processing. PMID- 2993242 TI - Regulation of CDP-diacylglycerol synthase activity in Saccharomyces cerevisiae. AB - The addition of ethanolamine or choline to inositol-containing growth medium resulted in a reduction of CTP:phosphatidate cytidylyltransferase (CDP diacylglycerol synthase; EC 2.7.7.41) activity in Saccharomyces cerevisiae. The reduction of activity did not occur in the absence of inositol. CDP diacylglycerol synthase activity was not regulated in a S. cerevisiae mutant strain (opi1; an inositol biosynthesis regulatory mutant) by the addition of phospholipid precursors to the growth medium. PMID- 2993244 TI - Regions on plasmid pCU1 required for the killing of Klebsiella pneumoniae. AB - Plasmid pCU1 was Kik+ (promotes killing of Klebsiella pneumoniae). All Tn5 insertions within the tra region of pCU1 were Kik-. Two other regions, kikA and kikB, were needed. They may be separated on different plasmids, but both must be mobilized into Klebsiella pneumoniae. Establishment of one kik region in K. pneumoniae followed by receipt of the second did not lead to killing. Kik was therefore intracellular and required concerted and transient action of both regions. PMID- 2993243 TI - Catabolite repression and role of cyclic AMP in CO2 fixation and H2 metabolism in Rhizobium spp. AB - CO2 fixation in Rhizobium meliloti was repressed by a variety of organic carbon sources. Cellular cyclic AMP levels were similar in repressed and nonrepressed cultures. Exogenous cyclic AMP or additional copies of the adenyl cyclase gene in cells experiencing repression failed to affect the rates of CO2 fixation. However, in R. japonicum catabolite repression of H2 utilization was partially circumvented by the presence of the R. meliloti adenyl cyclase gene. PMID- 2993245 TI - Nucleotide sequence and promoter region for the neutral protease gene from Bacillus stearothermophilus. AB - The thermostable neutral protease gene nprT of Bacillus stearothermophilus was sequenced. The DNA sequence revealed only one large open reading frame, composed of 1,644 bases and 548 amino acid residues. A Shine-Dalgarno sequence was found 9 bases upstream from the translation start site (ATG), and the deduced amino acid sequence contained a signal sequence in its amino-terminal region. The sequence of the first 14 amino acids of purified extracellular protease completely matched that deduced from the DNA sequence starting at GTC (Val), 687 bases (229 amino acids) downstream from ATG. This suggests that the protease is translated as a longer polypeptide. The amino acid sequence of the extracellular form of this protease (319 amino acids) was highly homologous to that of the thermostable neutral protease from Bacillus thermoproteolyticus but less homologous to the thermolabile neutral protease from Bacillus subtilis. A promoter region determined by S1 nuclease mapping (TTTTCC for the -35 region and TATTTT for the 10 region) was different from the conserved promoter sequences recognized by the known or factors in bacilli. However, it was very homologous to the promoter sequence of the spo0B gene from B. subtilis. The guanine-plus-cytosine content of the coding region of the nprT gene was 58 mol%, while that of the third letter of the codons was much higher (72 mol%). PMID- 2993246 TI - Presence of DNA, encoding parts of bacteriophage tail fiber genes, in the chromosome of Escherichia coli K-12. AB - The classical T-even bacteriophages recognize host cells with their long tail fibers. Gene products 35, 36, and 37 constitute the distal moiety of these fibers. The free ends of the tail fibers, which are formed by the CO2H terminus of gene product 37, possess the host range determinants. It was found that 4 out of 10 different strains of Escherichia coli K-12 contained regions of chromosomal DNA which hybridized with a probe consisting of genes 35, 36, and 37 of the T even phage K3. From one strain this homologous DNA, which was associated with an EcoRI fragment of about 5 kilobases, was cloned into plasmid pUC8. Two independently recovered hybrid plasmids had undergone a peculiar rearrangement which resulted in the loss of about 3 kilobases of cloned DNA and a duplication of both the vector and the remaining chromosomal DNA. The mechanisms causing this duplication-deletion may be related to that of transposases. The cloned DNA was capable of recombination with phage T4 gene 36 and a phage T2 gene 37 amber mutant. DNA sequencing revealed the existence of regions of identity between the cloned DNA and genes 36 and 37 of phage T2. In addition, after growth of a derivative of phage K3 on a strain harboring T2 DNA, it was found that this phage contained the same parts of the T2 tail fiber genes which had been recovered from the bacterial chromosome. There appears to be little doubt that the phage had picked up this DNA from the host. The possibility is considered that a repertoire of parts of genes 36 and 37 of various T-even-type phages is present in their hosts, allowing the former to change their host ranges. PMID- 2993248 TI - Synthesis, export, and assembly of Aeromonas salmonicida A-layer analyzed by transposon mutagenesis. AB - Suicide plasmid pJB4JI, containing transposon Tn5 and phage Mu, was introduced into Aeromonas salmonicida 449 which produces a surface protein array known as the A-layer. Kanamycin-resistant exconjugants of 449 with altered ability to produce the A-layer were selected by virtue of their altered colonial morphology and color on medium containing the dye Congo red. Analysis of culture supernatants, periplasmic shock fluid, outer membranes, and whole-cell lysates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with a monoclonal antibody to A-protein revealed five classes of single-insertion mutations that affected the ability of cells to produce and export A-protein and to assemble the A-layer. These studies suggest that A-protein is produced from a single chromosomal gene. The subunits subsequently pass through the periplasm and across the outer membrane. At least one gene product is required for this export. Assembly of A-layer on the cell surface then requires the presence of O polysaccharide chains on the lipopolysaccharide. In one case, insertion of Tn5 resulted in loss of ability to produce both A-protein and lipopolysaccharide with O polysaccharide chains, suggesting that synthesis of A-protein and synthesis of lipopolysaccharide may involve coordinate regulation. PMID- 2993247 TI - Determination of the transcription initiation site and identification of the protein product of the regulatory gene xylR for xyl operons on the TOL plasmid. AB - The xylR gene is a regulatory gene on the TOL plasmid, which acts in a positive manner on xyl operons for degradation of toluene and xylenes in Pseudomonas putida. A DNA fragment containing the xylR promoter region was cloned on promoter probing vectors, and its nucleotide sequence was determined. The transcription initiation site of the xylR gene was determined in cells of P. putida and Escherichia coli by S1 nuclease and reverse transcriptase mapping. Two initiation sites were detected which were identical in both P. putida and E. coli. The amounts of mRNA synthesized in both bacterial cells were almost the same and independent of the inducers for xyl operons. The consensus sequences for E. coli promoters were found in the region preceding the respective transcription initiation sites. The product of the xylR gene was identified by the maxicell system as a protein with an approximate molecular weight of 67,000. PMID- 2993249 TI - Plasmid-mediated mineralization of 4-chlorobiphenyl. AB - Strains of Alcaligenes and Acinetobacter spp. were isolated from a mixed culture already proven to be proficient at complete mineralization of monohalogenated biphenyls. These strains were shown to harbor a 35 X 10(6)-dalton plasmid mediating a complete pathway for 4-chlorobiphenyl (4CB) oxidation. Subsequent plasmid curing of these bacteria resulted in the abolishment of the 4CB mineralization phenotype and loss of even early 4CB metabolism by Acinetobacter spp. Reestablishment of the Alcaligenes plasmid, denoted pSS50, in the cured Acinetobacter spp. via filter surface mating resulted in the restoration of 4CB mineralization abilities. 4CB mineralization, however, proved to be an unstable characteristic in some subcultured strains. Such loss was not found to coincide with any detectable alteration in plasmid size. Cultures capable of complete mineralization, as well as those limited to partial metabolism of 4CB, produced 4 chlorobenzoate as a metabolite. Demonstration of mineralization of a purified 14C labeled chlorobenzoate showed it to be a true intermediate in 4CB mineralization. Unlike the mineralization capability, the ability to produce a metabolite has proven to be stable on subculture. These results indicate the occurrence of a novel plasmid, or evolved catabolic plasmid, that mediates the complete mineralization of 4CB. PMID- 2993250 TI - DNA sequence and complementation analysis of a mutation in the rplX gene from Escherichia coli leading to loss of ribosomal protein L24. AB - A mutation in Escherichia coli leads to the loss of ribosomal protein L24, severely impaired growth, and a temperature-sensitive phenotype. The mutation was shown to be in rplX, the gene for protein L24, and was due to the alteration of an AAA codon to a TAA stop codon at position 61 in rplX that resulted in a 20 amino acid peptide instead of the 104 amino acids of wild-type L24 protein. rplX genes from three temperature-resistant and fast growing pseudorevertants of the mutant were cloned and sequenced. They were found to have different base substitutions in the TAA codon, resulting in the reappearance of a full-sized protein L24 moiety. Complementation of the slow growth in trans could be achieved with several plasmids containing at least the spc promoter and intact L14 and L24 genes. Plasmids containing genes distal to rplX could further stimulate growth, and the wild type arose when the entire spc operon and the alpha operon were present. In all cases, protein L24 was expressed by the plasmids. Therefore, slow growth could be explained by polarity extending to the alpha operon. However, temperature sensitivity could not be complemented by any of the plasmids in trans, although we found that this phenotype was caused by the mutation in the rplX gene. PMID- 2993251 TI - Mu-lac insertion-directed mutagenesis in a pectate lyase gene of Erwinia chrysanthemi. AB - The pelC gene, which encodes one of the five major pectate lyase (PL) isoenzymes in Erwinia chrysanthemi 3937, designated PLc, was subcloned from a hybrid lambda phage into a pBR322 derivative and mutagenized with a mini-Mu-lacZ transposable element able to form fusions to the lacZ gene. One plasmid (pAD1) which had an inactivated pelC gene and a Lac+ phenotype was selected in Escherichia coli. This plasmid was introduced into Erwinia chrysanthemi, and the pelC::mini-Mu insertion was substituted for the chromosomal allele by homologous recombination. This strain lacks the PLc isoenzyme. This Erwinia chrysanthemi strain has a Lac+ phenotype that is inducible by polygalacturonate, as are the wild-type PL activities. PMID- 2993252 TI - Isolation and characterization of transposon Tn5-induced mutants of Pseudomonas perfectomarina defective in nitrous oxide respiration. AB - Transposon (Tn5) mutagenesis of Pseudomonas perfectomarina with the plasmid pSUP2021 [(pBR325-Mob(RP4))::Tn5] and the chromosomally integrated RP4 plasmid in Escherichia coli as the donor, produced three distinct groups of mutants that were defective in nitrous oxide respiration. One group of mutants lacked the structural protein of N2O reductase, the second synthesized a copper-free apoprotein; and a third group expressed a low level of intact enzyme. The mutants provided evidence for N2O being the immediate precursor of dinitrogen in denitrification and documented the essentiality of the copper enzyme. Synthesis of N2O reductase depended strongly on the growth conditions, with N2O-grown cells expressing the lowest level of enzyme. Regulatory responses of mutants elicited by nitrate or oxygen were unaltered when compared with wild-type behavior. PMID- 2993253 TI - Characterization of a membrane-regulated sugar phosphate phosphohydrolase from Lactobacillus casei. AB - One of the key components of the futile xylitol cycle of Lactobacillus casei Cl 16 is a phosphatase which dephosphorylates xylitol 5-phosphate to xylitol prior to the expulsion of the pentitol from cells. This enzyme has been partially purified and characterized. The phosphatase is active against a variety of four-, five-, and six-carbon sugars and sugar alcohols phosphorylated at the terminal 4, 5, and 6 positions, respectively, but exhibits little or no affinity for substrates phosphorylated at the C-1 position. The enzyme has an apparent molecular weight of 62,000 and a pH optimum between 5.5 and 6, and it requires a divalent cation (Mg2+) for maximal activity. A single protein band, exhibiting phosphatase activity, was excised from polyacrylamide gels and used to prepare antiphosphatase sera in rabbits. The antiserum was used to detect the enzyme on polyacrylamide gels and to determine the molecular weight of the monomer on sodium dodecyl sulfate-polyacrylamide gels. With a subunit molecular weight of 32,000, the native enzyme appears to be a dimer. Phosphatase activity and substrate specificity are regulated by some component associated with the cytoplasmic membrane. PMID- 2993255 TI - Therapeutic trial of plasma exchange in osteosclerotic myeloma associated with the POEMS syndrome. AB - A patient with osteosclerotic myeloma and POEMS syndrome, unresponsive to pulse prednisone and melphalan therapy, was admitted to the hospital for a trial of plasma exchange therapy. The presentation included IgG lambda monoclonal gammopathy, peripheral neuropathy, hepatosplenomegaly, hyperpigmentation and thickening of the skin, edema, and tense ascites. Laboratory tests confirmed hypothyroidism, hypogonadism, and adrenal insufficiency. Six exchange procedures failed to affect the clinical course, and the patient died. Greater-than-one plasma-volume exchanges (patient's measured plasma volume, 2,703 cc) were performed. When IgG and cholesterol removal were compared to the predicted removal, based on the volume of plasma removed, significantly less reduction in concentration than predicted was measured. IgG concentrations increased postapheresis and, at 2 weeks, three-fourths of the removed IgG had reaccumulated. A reduced efficiency of removal of both IgG and cholesterol can be explained by postulating increased vascular permeability with free exchange of soluble substances from one compartment to another. If an abnormal product is produced by the disease and is responsible for the clinical syndrome, a more intensive schedule of plasma exchange therapy may be needed to achieve a sustained depletion of the responsible soluble substance. Alternatively, neither increased vascular permeability or the clinical manifestations are responsive to removal of a soluble substance or are caused by a soluble substance produced by the malignancy. PMID- 2993254 TI - Phage-mediated cloning of bldA, a region involved in Streptomyces coelicolor morphological development, and its analysis by genetic complementation. AB - Streptomyces coelicolor bald (bld) mutants form colonies of vegetative substrate mycelium, but do not develop aerial hyphae or spore chains. The bldA strains form none of the four antibiotics known to be produced by the parent strain. With a vector derived from the temperate bacteriophage phi C31, a 5.6-kilobase fragment of wildtype DNA was cloned which restored sporulation to five independent bldA mutants when lysogenized with the recombinant phage. The cloned gene(s) was dominant over the mutant alleles. Phage integration by recombination of the cloned bldA+ DNA with the bldA region of each mutant produced mainly sporulating colonies, presumably heterozygous bldA+/bldA partial diploids for the insert DNA. However, a minority of these primary transductants were bald and were apparently homozygous bldA/bldA mutant partial diploids, formed by some homogenetization process. The phages released from the bald lysogens carried bldA mutations and were used to show that bldA+ sequences had been cloned and that fine mapping of the region could be performed. PMID- 2993256 TI - Purification and properties of thiosulfate reductase from Desulfovibrio vulgaris, Miyazaki F. AB - Thiosulfate reductase was purified to an almost homogeneous state from Desulfovibrio vulgaris, strain Miyazaki F, by ammonium sulfate precipitation, chromatography on DEAE-Toyopearl, Ultrogel AcA 34, and hydroxylapatite, and disc electrophoresis. The specific activity was increased 580-fold over the crude extract. The molecular weight was determined by gel filtration to be 85,000 89,000, differing from those reported for thiosulfate reductases from other Desulfovibrio strains. The enzyme had no subunit structure. When coupled with hydrogenase and methyl viologen, it stoichiometrically reduced thiosulfate to sulfite and sulfide with consumption of hydrogen. It did not reduce sulfite or trithionate. Cytochrome c3 was active as an electron donor. More than 0.75 mM thiosulfate inhibited the enzyme activity. o-Phenanthroline and 2,2'-bipyridine inhibited the enzyme and ferrous ion stimulated the reaction. PMID- 2993259 TI - Amino acid sequence of acylphosphatase from porcine skeletal muscle. AB - The amino acid sequence of acylphosphatase from porcine skeletal muscle was determined. It consists of 98 amino acid residues with N-acetylserine at the amino (N)-terminus: Ac-Ser-Thr-Ala-Arg-Pro-Leu-Lys-Ser-Val-Asp-Tyr-Glu-Val-Phe Gly -Arg-Val-Gln-Gly-Val-Cys-Phe-Arg-Met-Tyr-Thr-Glu-Asp-Glu-Ala-Arg-Lys-Ile -Gly Val-Val-Gly-Trp-Val-Lys-Asn-Thr-Ser-Lys-Gly-Thr-Val-Thr-Gly-Gln -Val-Gln-Gly-Pro Glu-Glu-Lys-Val-Asn-Ser-Met-Lys-Ser-Trp-Leu-Ser-Lys -Ile-Gly-Ser-Pro-Ser-Ser-Arg Ile-Asp-Arg-Thr-Asn-Phe-Ser-Asn-Glu-Lys- Thr-Ile-Ser-Lys-Leu-Glu-Tyr-Ser-Asn-Phe Ser-Ile-Arg-Tyr-OH. This sequence has three substitutions of amino acid residues, i.e., Thr/Ala, Ile/Val, and Ile/Val at positions 26, 68, and 96, respectively, from that of horse muscle acylphosphatase, formerly the only mammalian acylphosphatase with known sequence. PMID- 2993258 TI - A compilation of analytical data from inhibition studies on DNA polymerases and some of its implications. AB - In connection with the characterization of two DNA polymerases (DPols) of Chlorella, we have extensively surveyed the literature on inhibition studies on DPols in various eukaryotes. By applying Tamiya's plot (1), we have obtained two parameters for each of the inhibitors, phi- and n-values, which express the enzyme sensitivity to the drug and the number of inhibitor molecules present in the enzyme-inhibitor complex that is principally involved in the inhibition, respectively. By inspecting these parameters for the three mammalian DPols, alpha , beta-, and gamma-pols, as well as other eukaryotic DPols, we have found that: [1] inhibitors commonly utilized for characterizing various DPols can be classified into two major groups, each having two subgroups, on the basis of a comparison of the phi values among alpha-, beta-, and gamma-pols. Moreover, the grouping seems not to be merely coincidental, but to be intrinsically related to facets of the enzyme reaction, which may be taken to reflect evolutionary differences in DPol structure and function among the three DPols; [2] the remarkable n value, n = 1/2, that has been found for the inhibitors competitive with dCTP in Chlorella DPols has also been detected widely in many other eukaryotic DPols. Based on the first finding as well as many other data on various DPols, we have proposed an evolutionary scenario for eukaryotic DPols. Based on the second finding, we have hypothesized a novel role for dCTP as a cofactor, probably an apparent allosteric effector, in the nucleotide transfer reaction mechanism. PMID- 2993257 TI - Ouabain-resistant cells cultured in a synthetic medium. AB - Several strains of kidney and liver cells cultured in a synthetic medium were found to be resistant to ouabain. These cell strains were characterized because this resistance may serve as a good marker in genetic studies on somatic cells in chemically defined conditions in the absence of Na+ related growth factors and hormones. The phenotype was stable in the absence of selection for at least two years, and the original strains before adaptation to the synthetic medium were found to have ouabain sensitivity equal to the corresponding cells in the synthetic medium. The resting membrane potential, Na+,K+-ATPase activity, and growth rate of the resistant cells were similar to those of ouabain-sensitive cells. The resistance of the cells was not affected by serum or antibodies against some cytoskeletal proteins and the sensitivity of the Na+,K+-ATPase was not restored by partial purification of the membranes. Western blotting of the Na+,K+-ATPase of the ouabain-resistant cells showed that the molecular weights of its two subunits and its immunoreactivity were similar to those of the enzyme from the ouabain-sensitive strain. Thus the ouabain resistance is caused not by ouabain-like hormone produced by the cells or change in the cytoskeletal system, but by a mutation resulting in expression of an ouabain-resistant ATPase gene. PMID- 2993260 TI - Amino acid sequence of acylphosphatase from rabbit skeletal muscle. AB - The complete amino acid sequence of acylphosphatase from rabbit skeletal muscle has been elucidated by automatic Edman degradation of peptides obtained from staphylococcal protease and trypsin digestions. The enzyme consisted of a single polypeptide chain of 98 amino acid residues, lacking only histidine. Its amino (N)-terminus was blocked by an acetyl group. The presented sequence of rabbit muscle enzyme was compared with those of equine and porcine muscle enzymes. There were four unique replacements, i.e., Arg-4, Asp-28, Arg-31, and Glu-56 in the sequences of both equine and porcine muscle enzymes were replaced by Gly, Gly, Lys, and Asp, respectively, in that of rabbit muscle enzyme. Extensive structural homology was observed among the three enzymes. PMID- 2993261 TI - Gangliosides as paramyxovirus receptor. Structural requirement of sialo oligosaccharides in receptors for hemagglutinating virus of Japan (Sendai virus) and Newcastle disease virus. AB - A sensitive assay system for receptor activity of gangliosides to paramyxovirus was developed. This system involves incorporation of gangliosides into neuraminidase-treated chicken erythrocytes (asialoerythrocytes) followed by estimation of virus-mediated agglutination and hemolysis. The asialoerythrocytes coated with I-active ganglioside (Sia alpha 2-3Gal beta 1-4GlcNAc beta 1-3(Gal alpha 1-3Gal beta 1-4GlcNAc beta 1-6)Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer) were effectively agglutinated by hemagglutinating virus of Japan (HVJ, Sendai virus). The hemolysis of the asialoerythrocytes mediated by HVJ was restored to the highest level by labeling the cells with gangliosides possessing lacto-series oligosaccharide chains, i.e., I-active ganglioside, N acetylneuraminosylparagloboside (SiaPG(NeuAc)), and i-active ganglioside (Sia alpha 2-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-Cer). The specific receptor activity of ganglioside GD1a possessing a gangliotetraose chain was lower than those of the gangliosides described above. Gangliosides GM3, GD3, GM1a, GD1b, SiaPG(NeuGc) showed little effect on the restoration of HVJ-mediated hemolysis. On infection with Newcastle disease virus (NDV), the highest specific restoration of lysis was found in chicken asialoerythrocytes coated with SiaPG(NeuAc or NeuGc) and GM3(NeuAc or NeuGc), whereas those coated with I-active ganglioside, GD3, GM1a, and GD1b showed very low NDV-mediated hemolysis. The above results indicate that the determinants of receptor for HVJ contain sialylated branched and/or linear lacto-series oligosaccharides carried by I,i-active gangliosides and SiaPG(NeuAc) and sialosylgangliotetraose chain carried by GD1a. The determinants for NDV are carried by SiaPG(NeuAc or NeuGc) containing linear lacto-series oligosaccharide and GM3(NeuAc or NeuGc). The absence of detectable binding of free oligosaccharides obtained from I-active ganglioside and sialoglycoprotein GP-2 isolated from bovine erythrocyte membranes as HVJ receptor (Suzuki, Y., et al. J. Biochem. (1983) 93, 1621-1633; (1984) 95, 1193-1200) indicates that HVJ recognizes the sialooligosaccharides oriented out of the lipid bilayer in the cell membranes where the hydrophobic ceramide or peptide backbone of the receptor is integrated. PMID- 2993262 TI - Absence of phosphatidylinositol phosphodiesterase in the head of a Drosophila visual mutant, norpA (no receptor potential A). AB - Phosphatidylinositol phosphodiesterase activity was found to be entirely absent in the head of the Drosophila visual mutant, norpA. It was also demonstrated that the enzyme activity was highly concentrated in the retinular cells of a normal head. Furthermore, the enzyme was revealed to be in a membrane bound form. In view of the present results and our previous work on the reduction of diacylglycerol kinase activity in the norpA mutant, phosphatidylinositol metabolism may play an important role in the generation of photoreceptor potentials. PMID- 2993263 TI - Molecular cloning and nucleotide sequence of DNA complementary to rat ribosomal protein S26 messenger RNA. AB - A cDNA clone specific for rat ribosomal protein S26 was isolated by a positive hybridization translation assay from a cDNA library made for 8-9S poly(A)mRNA from regenerating rat liver. The nucleotide sequence of the cDNA was determined. The sequence contains 23 base pairs in the 5' noncoding region, 345 base pairs in the protein coding region and 67 base pairs in the 3' noncoding region besides the poly(A) tail. The primary structure of the protein S26 was deduced from the nucleotide sequence. It consists of 115 amino acids. Its molecular weight is 13,015 and its pI is about 11.1. The calculated amino acid composition is consistent with the reported composition of S26. From the results of Southern blot analysis, the protein S26 appears to have multiple genes. PMID- 2993264 TI - Phosphorus-31 nuclear magnetic resonance and electronic spectroscopic studies of adrenodoxin reductase and its binary complex with NADP+. AB - The 31P NMR spectra of NADPH-adrenodoxin reductase and its complex with NADP+ are reported. The spectrum of adrenodoxin reductase showed two doublets arising from the phosphorus nuclei in the pyrophosphate group of FAD. Both doublets were shifted upfield to different extents in comparison with those of free FAD. Further, one of the doublets of phosphorus nuclei of the pyrophosphate group of bound NADP+ in the complex of adrenodoxin reductase and NADP+ was considerably shifted upfield in comparison with that of free NADP+. The spectrum of the complex of the reductase and NADP+ showed that the resonance of the 2'-phosphate group of NADP+ bound to the reductase was shifted downfield by 1.37 ppm compared with that of free NADP+ in the dianionic state. The 2'-phosphate resonance of bound NADP+ was independent of pH within the physiological range, whereas that of free NADP+ changed according to its ionization. The resonance of the 2'-phosphate group of NADP+ bound to the reductase also revealed that the ratio for the complex of NADP+ and the reductase was 1:1, and that this complex formation was inhibited by a high KCl concentration. These results were confirmed by electronic spectroscopic studies. PMID- 2993265 TI - Mechanism of stimulation of globin synthesis by adenosine-3',5'-monophosphate and guanosine 5'-triphosphate in a rabbit reticulocyte lysate system. AB - The inhibition of globin synthesis in hemin-deficient rabbit reticulocyte lysates is due to the activation of a hemin-controlled translational inhibitor (HCI) that specifically phosphorylates eIF-2 alpha. High concentrations of cAMP (5-10 mM) and GTP (1-2 mM) stimulated the globin synthesis in hemin-deficient lysates when these compounds were added at the initial stage of incubation. The mechanism of the stimulation by cAMP and GTP was studied using hemin-deficient lysates, the N ethylmaleimide (NEM)-treated HCI-supplemented lysates and a partially purified initiation factor, eIF-2. As the stimulation of globin synthesis by these compounds must be due to the prevention of the inhibition of globin synthesis, or due to the restoration of globin synthesis, or both, the preventive and restorative effects of these compounds were examined. As for the preventive effect, it was observed that a) the activation of HCI in the postribosomal supernatant of reticulocytes was prevented by GTP, but not by cAMP, and b) cAMP and GTP inhibited the phosphorylation of eIF-2 alpha in hemin-deficient lysates. As for the restorative effect of cAMP and GTP, it was observed that c) these compounds restored the globin synthesis and the binding of [35S]Met-tRNAf to the 40S ribosomal subunits, and promoted the dephosphorylation of eIF-2(alpha P), d) the rates of the restored synthesis of globin were lower than the control, and e) cAMP promoted the release of [3H]GDP from the eIF-2(alpha P) X [3H]GDP complex and the formation of eIF-2(alpha P) X eIF-2B complex. Finding (d) indicates that steps involved in the restorative effect of these compounds may not contribute to the stimulation of the globin synthesis in hemin-deficient lysates. The data on the preventive and restorative effects of cAMP and GTP showed that these compounds affected multiple steps. That is, cAMP inhibited the phosphorylation of eIF-2 alpha and promoted both the release of GDP from eIF-2 and the formation of eIF-2(alpha P) X eIF-2B complex, and GTP prevented both the activation of HCI and the phosphorylation of eIF-2 alpha. Though cAMP and GTP affected multiple steps, it is suggested that cAMP stimulates the globin synthesis by inhibiting the phosphorylation of eIF-2 alpha and that GTP stimulates the globin synthesis chiefly by preventing the activation of HCI in hemin-deficient lysates. PMID- 2993266 TI - Effect of lipid composition on HVJ-mediated fusion of glycophorin liposomes to erythrocytes. AB - We previously reported that liposomes containing glycophorin or gangliosides, both of which were isolated from human erythrocytes, are efficiently fused to erythrocyte membranes in the presence of HVJ (Umeda, M. et al., J. Biochem. 94, 1955-1966 (1983), and Virology 133, 172-182 (1984]. In the present work, the effect of lipid composition in glycophorin liposomes on their sensitivity to fusion with erythrocytes was studied. Very little fusion occurred when glycophorin liposomes composed of dipalmitoylphosphatidylcholine-dicetylphosphate (9:1), dimyristoylphosphatidylcholine-dicetylphosphate (9:1), or egg yolk phosphatidylcholine-dicetylphosphate (9:1) were incubated with human erythrocytes in the presence of HVJ at 37 degrees C. Addition of cholesterol into these liposomal membranes greatly enhanced the sensitivity of the liposomes to fusion. The presence of phosphatidic acid and phosphatidylethanolamine in liposomes also enhanced the sensitivity, whereas the presence of lysophosphatidylcholine had no significant effect on the ability of the liposomes to fuse. The fusion efficiency of liposomes was also enhanced by the presence of glucosylceramide. Change of lipid composition in liposomes had, however, no appreciable influence on the HVJ mediated binding of liposomes to erythrocytes, suggesting that the interaction between HANA protein of HVJ and glycophorin in liposomes was not affected by the lipid composition of the liposomes. PMID- 2993267 TI - Hydrolysis of protamine by calcium-activated neutral protease (CANP). AB - To determine the substrate recognition mechanism in calcium-activated neutral protease (CANP), the hydrolytic velocities for some possible substrates were compared. In general, succinylated polypeptides were poorer substrates than unmodified ones, suggesting that CANP interacts with positively charged amino groups and/or repels negatively charged succinyl groups in substrates. Among the substrates examined, protamine was degraded quite rapidly in a restricted manner. This degradation of protamine was remarkably accelerated by the addition of salt, and, in the absence of salt, protamine was inhibitory as to the degradation of vimentin by CANP. Protamine was separated into components and the sites cleaved by CANP were determined. CANP cleaved the clupeine YII and Z components at two sites, both being arginyl-arginine bonds, and the amino acid sequences around these sites were almost identical between YII and Z. No other arginyl-arginine bond was cleaved at all. These results showed that CANP prefers basic amino acid side chains but its specificity is very restricted. PMID- 2993268 TI - Tetrahymena ubiquitin-histone conjugate uH2A. Isolation and structural analysis. AB - The ubiquitin-histone H2A conjugate, uH2A, of the protozoan Tetrahymena pyriformis was isolated by gel chromatography and octadecylsilyl-silica chromatography, from the fractions on the chromatographic purification of histone H2A [Fusauchi, Y. & Iwai, K. (1983) J. Biochem. 93, 1487-1497]. The uH2A showed an amino acid composition corresponding to the sum of an equimolar mixture of two protozoan H2A variants and protozoan free ubiquitin. N- and C-terminal sequencing of the uH2A, by Edman degradation and carboxypeptidase P digestion, showed a branched structure having two N-terminals, those of the H2A and ubiquitin components, and one C-terminal, that of each H2A variant component. Further structural analyses of the uH2A, by tryptic digestion of citraconylated uH2A and of a ubiquitinated BrCN fragment, showed that the ubiquitin C-terminal Gly-Gly is linked to the epsilon-amino group of either Lys-123, 125, or 126 in the H2A sequence, and that the ubiquitin sequence is similar to that of calf thymus but differs at least in the sequence of residues 12-27. The deducted structure was compared with the only known uH2A structure, that of calf thymus, with special reference to the branched site. PMID- 2993269 TI - Structural analysis of a developmentally regulated 25-kDa protein gene of Sarcophaga peregrina. AB - In the previous paper, we described the identification of two abundant mRNAs of Sarcophaga peregrina (flesh-fly) which are selectively expressed in the fat body of middle third instar larvae. One of these mRNAs was found to encode a protein with a molecular mass of about 25,000 (25-kDa protein) when translated in vitro (Tamura, H., et al. (1983) Dev. Biol. 99, 145-151). Present paper reports the nucleotide sequence of a 2.3 kb DNA containing the entire gene for the 25-kDa protein. This gene consisted of four exons and contained an open reading frame for 184 amino acids. A CAT box and a TATA box were found in the 5'-flanking sequence. A poly A addition signal of AATAAA was assigned to the non-coding region in the fourth exon. A sequence having 75% homology with SV40 enhancer core sequence was identified in the non-coding region of the first exon. PMID- 2993270 TI - Calcium-dependent interaction of actin filaments with actin binding protein in the presence of calmodulin and caldesmon. AB - Caldesmon, calmodulin-, and actin-binding protein of chicken gizzard did not affect the process of polymerization of actin induced by 0.1 M KCl. Caldesmon binds to F-actin, thus inhibiting the gelation action of actin binding protein (ABP; filamin). Low shear viscosity and flow birefringence measurements revealed that in a system of calmodulin, caldesmon, ABP, and F-actin, gelation occurs in the presence of micromolar Ca2+ concentrations, but not in the absence of Ca2+. Electron microscopic observations showed the Ca2+-dependent formation of actin bundles in this system. These results were interpreted by the flip-flop mechanism: in the presence of Ca2+, a calmodulin-caldesmon complex is released from actin filaments on which ABP exerts its gelating action. On the other hand, in the absence of Ca2+, caldesmon remains bound to actin filaments, thus preventing the action of ABP. PMID- 2993272 TI - Activation of guinea pig peritoneal macrophages by platelet activating factor (PAF) and its agonists. AB - The platelet activating factor (PAF: 1-O-alkyl-2-O-acetyl-sn-glycero-3 phosphocholine) and its analogs were examined to determine their effects on guinea pig peritoneal macrophages. PAF activated macrophages, but its effect on macrophages was much weaker than that observed on platelets: the concentration required for 50% maximum activation was 8.5 X 10(-6) M for macrophages and 2.9 X 10(-10) M for platelets. Three PAF agonists, 1-O-octadecyl-2-O-(N,N dimethylcarbamoyl)-glycero-3-phosphocholine (Compound I), 1-O-octadecyl-2 acetamido-2-deoxy-glycero-3-phosphocholine (Compound II), and 1-O-octadecyl-2-O methyl-glycero-3-phosphocholine (Compound III), showed higher activity in stimulating macrophage function than PAF. The abilities of these non metabolizable PAF agonists to activate macrophage paralleled their relative potency to induce platelet activation. The sn-3 enantiomers of PAF and Compound III exhibited activity, while the sn-1 did not. By comparing the activities of derivatives of Compound III, it was shown that the long-chain alkyl-ether group in the glycerol-1 position, a relatively small size of the substituent on the hydroxy group at the sn-2 position, and the choline moiety in the glycerol-3 position must play critical roles in the process of macrophage activation. A specific PAF antagonist, CV3988, which inhibits PAF-induced platelet activation and hypotension, inhibited the activation of macrophages caused by PAF and its agonists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993271 TI - Effect of prostaglandin E2 on PZ-peptidase and several other peptidase activities in a clonal osteoblast-like cell line derived from newborn mouse calvaria. AB - The effects of prostaglandin E2(PGE2) on the degradation of collagen and non collagenous peptides in clonal osteoblastic MC3T3-E1 cells were investigated by using highly sensitive assay methods for PZ-peptidase, collagenase-like peptidase (CL-peptidase), dipeptidyl-aminopeptidase (DAP), leucine aminopeptidase (LAP), and post-proline cleaving enzyme (PPCE). PGE2, at concentrations of 0.1 to 4.0 micrograms/ml, doubled the PZ-peptidase and CL-peptidase activities in the cells on 24 h culturing in a dose-dependent manner. PGE2, at a concentration of 2.0 micrograms/ml, enhanced the specific activities of PZ-peptidase, CL-peptidase, DAP, LAP, and PPCE for 75 h after the start of PGE2 stimulation. The time dependent changes in PZ-peptidase and CL-peptidase activities showed similar patterns, and 3- and 2-fold increases were seen after 48 h, respectively. The protein and DNA contents gradually increased after addition of PGE2. Since the PZ peptidase and CL-peptidase, involved in degradation of collagen peptides, were significantly induced by PGE2 in comparison with LAP and PPCE, involved in the degradation of non-collagenous peptides, these results show that PGE2 specifically stimulates induction of collagen catabolizing enzymes in clonal osteoblasts. PMID- 2993273 TI - Analyses of muscle proteins in a patient with a mitochondrial myopathy. AB - Using the small amounts of muscle available from biopsy (approximately 100 mg), from both normal controls and a patient with a previously identified defect of the mitochondrial electron transfer protein complex III, we analyzed both structural and mitochondrial proteins. The myosin light chains were found to be unchanged with respect to charge or size between patient and control. Two prominent proteins detected after two dimensional gel electrophoresis were present in the patient's total homogenised muscle protein but were not detected in the controls. One protein was positively identified as cytochrome c oxidase subunit II and the other tentatively as a component of the ATP synthetase. We suggest that the increased amounts of these proteins represents a response of the patients muscle cells to the ATP deficiency caused by the primary lesion in complex III. PMID- 2993274 TI - Partial purification of the apolipoprotein E receptor of rat liver membrane. AB - The liver has two distinct lipoprotein receptors; one is the apolipoprotein (apo) B, E receptor and the other the apo-E receptor. In this study, the protein to which apo-E HDLc (cholesterol-induced high density lipoprotein containing apo-E as the predominant protein species) bound specifically was partially purified from the rat liver membrane by ion exchange chromatography and preparative gel electrophoresis. The molecular weight of the protein was estimated to be about 36K daltons and the protein bound to 125I-apo-E HDLc in a specific and saturable manner, suggesting that the protein is the apo-E receptor. PMID- 2993275 TI - The iron-sulfur protein of cytochrome bc1 complex. Its occurrence in the mitochondrial inner membrane in excess of the amount constituting the complex. AB - Radioimmunoassay and quantitative immunoblot analysis have been developed for quantitation of the iron-sulfur protein of cytochrome bc1 complex in order to compare its content in isolated cytochrome bc1 complex with that in electron transport particles. The result by radioimmunoassay indicated that the content of the iron-sulfur protein/mol of cytochrome b is higher by approximately 30%, on the average, in electron transport particles than in cytochrome bc1 complex. This observation was supported by the data of immunoblot analysis. Since approximately 1/3 of cytochrome b in electron transport particles is not attributed to cytochrome bc1 complex, but to succinate-ubiquinone oxidoreductase complex (Davis, K.A., Hatefi, Y., Poff, K. L., and Butler, W. L. (1973) Biochim. Biophys. Acta 325, 341-356), the ratio of the iron-sulfur protein detectable by radioimmunoassay in electron transport particles to that in cytochrome bc1 complex is calculated to be approximately 2 on the basis of the content of 2 mol of b-type heme/mol of the complex. Therefore, it appears that the mitochondrial inner membrane contains approximately two times as much of the immunoreactive iron-sulfur protein as what is expected from the stoichiometry of one iron-sulfur center and two b-type hemes for cytochrome bc1 complex. This finding affords an interesting aspect in the study of biogenesis of cytochrome bc1 complex. PMID- 2993276 TI - Replication of the vesicular stomatitis virus genome in permissive and nonpermissive host cells. AB - Permissive infections of BHK cells and nonpermissive infections of Raji cells were probed for the accumulation of vesicular stomatitis virus intracellular RNAs. In Raji cells, the onset of vesicular stomatitis virus transcription and replication was delayed when compared to BHK cells, and the accumulation of plus and minus sense leader RNAs was significantly reduced. In contrast, full length plus and minus strand replicative RNAs accumulated in Raji cells to levels approximately equivalent to those in BHK cells. In both cell types, approximately four times as many minus strands as plus strands were detected late in the infections. At 16 h postinfection, 12% of the total genomic RNA synthesized in BHK cells was packaged and released whereas only 0.8% was released from Raji cells. In addition, of those particles released by Raji cells, only 1% were infectious whereas 77% of those released by BHK cells were infectious. The virions released from both cell types contained similar amounts of the five viral proteins, however. Analysis of virions from Raji cells revealed a faster electrophoretic mobility of the glycoprotein than the glycoprotein in virions released from BHK cells. These results suggest that Raji cells may be restricted in their ability to support a complete infection at the level of virus assembly rather than at the level of RNA replication. PMID- 2993277 TI - Kinase FA-mediated regulation of rabbit skeletal muscle protein phosphatase. Reversible phosphorylation of the modulator subunit. AB - A mechanism of activation of the ATP.Mg-dependent protein phosphatase (FC.M) has been proposed (Jurgensen, S., Shacter, E., Huang, C. Y., Chock, P. B., Yang, S. D., Vandenheede, J. R., and Merlevede, W. (1984) J. Biol. Chem. 259, 5864-5870) in which a transient phosphorylation by the kinase FA of the modulator subunit (M) is the driving force for the transition of the inactive catalytic subunit (FC) into its active conformation. Incubation of FC.M with kinase FA and Mg2+ and adenosine 5'-(gamma-thio)triphosphate results in thiophosphorylation of M and also a conformational change in the phosphatase catalytic subunit; however, the enzyme remains inactive. Proteolysis of this inactive, thiophosphorylated complex causes proteolytic destruction of the modulator subunit and yields an active phosphorylase phosphatase species. Similar treatment of the native inactive enzyme does not yield active phosphatase. Evidence is presented, suggesting that a molecule of modulator is bound at an "inhibitory site" on the native enzyme. This modulator does not prevent the conformational change in the phosphatase catalytic subunit upon incubation with kinase FA and ATP.Mg but does partially inhibit the expression of the phosphorylase phosphatase activity. PMID- 2993278 TI - Inhibition of fibrinogen receptor expression and serotonin release by leupeptin and antipain. AB - Human platelet agonists such as thrombin, ADP, and collagen stimulate the rapid expression of fibrinogen receptors. In other cell types, calcium-activated proteases have been suggested to participate in the mechanism of expression of cell surface receptors (Lynch, G., and Baudry, M. (1984) Science 224, 1057-1063). In platelets the majority of the neutral protease activity is calcium-activated protease. We examined the effects of leupeptin and antipain, two calcium activated protease inhibitors, on the expression of platelet fibrinogen receptors. These inhibitors abolished thrombin and ADP-induced fibrinogen binding. This inhibition required the addition of leupeptin or antipain prior to the agonist and was not due to displacement of fibrinogen from its receptor or inhibition of agonist binding to platelets. Leupeptin and antipain also inhibited fibrinogen-independent thrombin-stimulated release of serotonin. These results are discussed in relation to the involvement of calcium-activated protease in early events of platelet activation. PMID- 2993279 TI - Evidence for tight coupling of thyrotropin-releasing hormone receptors to stimulated inositol trisphosphate formation in rat pituitary cells. AB - The effect of decreasing the concentration of receptors for thyrotropin-releasing hormone (TRH) on the surface of cloned rat pituitary (GH3) cells on TRH stimulated inositol trisphosphate (Ins-P3) formation was investigated. Incubation of cells with dibutyryl cAMP (Bt2cAMP) for 16 h caused a decrease in [3H] TRH binding to intact cells to a minimum level 37 +/- 9.1% of control. Scatchard analysis of the concentration dependency of [3H]TRH binding showed that the effect of Bt2cAMP was to lower the receptor concentration without affecting its affinity for TRH. Similar decreases in [3H]TRH binding were found in cells incubated with 8-bromo-cAMP, cholera toxin, and sodium butyrate and, as shown previously, with TRH. In cells incubated with 1 mM Bt2cAMP for 16 h, but not for 1 h, the maximum TRH-induced increase in Ins-P3 was inhibited to 25 +/- 3.2% of that in control cells. Inhibition of TRH-induced Ins-P3 formation was also observed in cells treated with 8-bromo-cAMP, cholera toxin, and sodium butyrate for 16 h, and with TRH for 48 h. Inhibition of TRH-induced Ins-P3 formation and lowering of TRH receptor concentration caused by Bt2cAMP occurred in parallel with increasing doses of Bt2cAMP; at 16 h of exposure, half-maximal effects occurred with 0.3 mM Bt2cAMP. The concentration dependency of TRH-induced Ins-P3 formation was the same in control and Bt2cAMP-treated cells; half-maximal effects occurred with 10 nM TRH. These data demonstrate that decreases in TRH receptor concentration caused by several agents that act via different mechanisms are associated with reduced stimulation of Ins-P3 formation and suggest that the TRH receptor is tightly coupled to stimulation of hydrolysis of phosphatidylinositol 4,5-bisphosphate by a phospholipase C. PMID- 2993281 TI - Spectroscopic studies on the iron-sulfur centers of milk xanthine oxidase. AB - The optical electron paramagnetic resonance and Mossbauer spectral properties of the two iron-sulfur centers present in milk xanthine oxidase have been reexamined. It is found in the case of the optical spectral change observed on reduction of the enzyme that the two centers contribute approximately equally, with a ratio of spectral contributions for Fe/S I and Fe/S II of 0.55:0.45. This conclusion is based both on the behavior of the spectral change at wavelengths where only the two iron-sulfur centers contribute to the spectral change (under experimental conditions minimizing the effect of flavin semiquinone) during reductive titrations and a comparison of the spectra of 1- and 2-electron reduced enzyme under different conditions. This very similar spectral weighting for the two centers applies throughout the visible region. In the case of the EPR spectra, it is found from computer simulation of the signals observed under nonsaturating conditions that iron-sulfur center II exhibits g values of 1.902, 1.991, and 2.110 and does not exhibit two g values above that for the free electron, as has been reported (Lowe, J., Lynden-Bell, R.M., and Bray, R. C. (1972) Biochem. J. 130, 239-249). The g values for iron-sulfur center I obtained from the simulations are 1.894, 1.932, and 2.022. Finally, Mossbauer spectra of xanthine oxidase have been obtained, and it is found that while the two iron sulfur centers are indistinguishable in the oxidized state, the ferrous iron in one of the reduced iron-sulfur centers exhibits an unusually large quadrupole coupling. PMID- 2993280 TI - Enhanced transcription of fibroin gene in vitro on covalently closed circular templates. AB - A modified method for S1 nuclease assay has been developed to quantitate an amount of radioactive transcripts in the presence of unlabeled homologous RNA. Using this method, we have assayed in vitro transcription of fibroin gene on circular and linear templates. In a posterior silk gland cell extract, covalently closed circular (ccc) DNA forms the superhelical state and supports three to 10 times more transcription than nicked circular or linear DNA does. The efficient transcription on ccc template is also observed in a middle silk gland cell extract but never seen in a HeLa cell extract. When the posterior silk gland cell extract is first incubated with pBR322 DNA and then challenged for the fibroin gene transcription on ccc DNA, the pBR322 DNA of the ccc form, but not of the linear one, inhibits the reaction. A deletion mutant carrying only 44 base pairs of the 5'-flanking sequence reserves the preference of the ccc template, but seven single-point mutants at the TATA box and -20 region abolish it. These results suggest that some components in the silk gland extract supercoil ccc DNA and stimulate the fibroin gene transcription in the absence of the upstream enhancing sequence. PMID- 2993282 TI - Metabolism of the carbocyclic analogue of (E)-5-(2-iodovinyl)-2'-deoxyuridine in herpes simplex virus-infected cells. Incorporation of C-IVDU into DNA. AB - The carbocyclic analogues of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) and (E) 5-(2-iodovinyl)-2'-deoxyuridine (IVDU), in which the sugar moiety is replaced by a cyclopentane ring and which have been designated as C-BVDU and C-IVDU, respectively, are, like their parent compounds BVDU and IVDU, potent and selective inhibitors of herpes simplex virus type 1 (HSV-1) and, to a lesser extent, herpes simplex virus type 2 (HSV-2) replication. We have now synthesized the radiolabeled C-IVDU analogue, C-[125I]IVDU, and determined its metabolism by HSV-infected and mock-infected Vero cells. C-[125I]IVDU was effectively phosphorylated by HSV-1-infected cells and, to a lesser extent, HSV-2-infected cells. C-[125I]IVDU was not phosphorylated to an appreciable extent by either mock-infected cells or cells that had been infected with a thymidine kinase deficient mutant of HSV-1. Furthermore, C-[125I]IVDU was incorporated into both viral and cellular DNA of HSV-1-infected Vero cells. This finding represents the first demonstration of the incorporation of a cyclopentylpyrimidine into DNA. PMID- 2993284 TI - The porcine LH/hCG receptor. Characterization and purification. AB - Porcine luteal LH/hCG receptor (LH/hCG R) was solubilized with 70-80% recovery from the crude plasma membrane fraction by Triton X-100 in the presence of 25% glycerol and protease inhibitors. The solubilized receptor maintained 90% of original activity at -60 degrees C for 90 days. Equilibrium association constant (Ka) values of 1.92, 2.22, and 2.03 X 10(10) M-1 were observed for the whole homogenate, plasma membrane fraction, and solubilized LH/hCG R preparations, respectively. The specific binding capacity for the same fractions were 49, 70, 55 fmol/mg protein, respectively. Complexes of LH/hCG R and Triton X-100 were resolved into two components with approximate Mr = 2.7 X 10(5) and 5.4 X 10(5) by gel filtration on Sepharose 6B and two glycoprotein components by chromatography on concanavalin A-Sepharose. Solubilized porcine LH/hCG R was purified by two cycles of affinity chromatography on highly purified hCG-Sepharose with an overall recovery of 30-35% of the initial activity in the Triton extract. Purified porcine LH/hCG R had a specific binding capacity of 2300 pmol/mg protein and a Ka = 1.5 X 10(10) M-1. Silver staining of sodium dodecyl sulfate polyacrylamide gel electrophoresis gels demonstrated that the major protein in porcine LH/hCG R preparations has Mr = 68,000. A weakly staining band at Mr = 45,000 was also observed in the purified receptor preparation. Analysis of iodinated purified LH/hCG R by autoradiography has confirmed these results. Porcine LH/hCG R was purified 40,000-fold by this method. PMID- 2993283 TI - Mitoxantrone and ametantrone inhibit hydroperoxide-dependent initiation and propagation reactions in fatty acid peroxidation. AB - The anthracenedione antineoplastic agents mitoxantrone and ametantrone are potent inhibitors of basal and drug-stimulated lipid peroxidation in a variety of subcellular systems (Kharasch, E. D., and Novak, R. F. (1983) J. Pharmacol. Exp. Ther. 226, 500-506). The mechanism by which these compounds function as antioxidants has been investigated using enzymic and chemical systems. Mitoxantrone and ametantrone inhibited NADPH-cytochrome P-450 reductase- and xanthine oxidase-catalyzed conjugated diene formation from linoleic acid in a concentration-dependent manner with half-maximal inhibition achieved at approximately 0.5 microM anthracenedione. Inhibition of linoleic acid peroxidation was not attributable to a decrease in P-450 reductase activity, hydroxyl radical scavenging, or iron chelation by the anthracenediones. Nonenzymic fatty acid peroxidation was also inhibited by the anthracenediones. Linoleic acid oxidation initiated by superoxide (ferrous iron autoxidation) or by hydroxyl radicals (Fenton's reagent) was diminished by mitoxantrone and ametantrone after a brief delay, suggesting an effect subsequent to activated oxygen-dependent initiation. In contrast, linoleic acid oxidation initiated by iron-dependent hydroperoxide decomposition was inhibited immediately. Reinitiation of linoleic acid oxidation in an anthracenedione-inhibited system was accomplished only by superoxide generation, but not by fatty acid hydroperoxide decomposition. These results suggest the anthracenediones diminished neither oxygen radical formation nor oxygen radical-dependent initiation of peroxidation. Rather, inhibition of fatty acid peroxidation by mitoxantrone and ametantrone results from the inhibition of hydroperoxide dependent initiation and propagation reactions. PMID- 2993285 TI - Reconstitution of the purified platelet fibrinogen receptor. Fibrinogen binding properties of the glycoprotein IIb-IIIa complex. AB - Several lines of evidence indicate that the platelet membrane glycoprotein IIb IIIa complex (GP IIb-IIIa) is necessary for the expression of platelet fibrinogen receptors. The purpose of the present study was to determine whether purified GP IIb-IIIa retains the properties of the fibrinogen receptor on platelets. Glycoprotein IIb-IIIa was incorporated by detergent dialysis into phospholipid vesicles composed of 30% phosphatidylcholine and 70% phosphatidylserine. 125I Fibrinogen binding to the GP IIb-IIIa vesicles, as measured by filtration, had many of the characteristics of 125I-fibrinogen binding to whole platelets or isolated platelet plasma membranes: binding was specific, saturable, reversible, time dependent, and Ca2+ dependent. The apparent dissociation constant for 125I fibrinogen binding to GP IIb-IIIa vesicles was 15 nM, and the maximal binding capacity was 0.1 mol of 125I-fibrinogen/mol of GP IIb-IIIa. 125I-Fibrinogen binding was inhibited by amino sugars, the GP IIb and/or IIIa monoclonal antibody 10E5, and the decapeptide from the carboxyl terminus of the fibrinogen gamma chain. Furthermore, little or no 125I-fibrinogen bound to phospholipid vesicles lacking protein or containing proteins other than GP IIb-IIIa (i.e. bacteriorhodopsin, apolipoprotein A-I, or glycophorin). Also, other 125I-labeled plasma proteins (transferrin, orosomucoid) did not bind to the GP IIb-IIIa vesicles. These results demonstrate that GP IIb-IIIa contains the platelet fibrinogen receptor. PMID- 2993286 TI - Characterization of mammalian neurofilament triplet proteins. Subunit stoichiometry and morphology of native and reconstituted filaments. AB - The three major proteins of mammalian neurofilaments of molecular weights 179,000 (NF1), 129,000 (NF2), and 66,500 (NF3) have been purified to homogeneity by multiple anion-exchange and hydroxylapatite absorption chromatography in 8 M urea. Silver staining of polyacrylamide gels of the purified proteins show single bands. In order to gain further insight into the molecular organization of the neurofilament triplet proteins, the molar stoichiometries and morphologies of native and reconstituted filaments and those isolated from developing brain were studied. Denaturing polyacrylamide gel electrophoresis followed by quantitative dye-binding analysis shows that the molar ratio of the three components in neurofilaments isolated from bovine spinal cord myelinated nerve is 4:2:1 (NF3:NF2:NF1). Comparison of the molar ratios of each component in neurofilaments isolated from rat, bovine, and human brain shows a variation in the ratio of each of these polypeptides and raises questions about the physiological uniqueness of the molar composition of the neurofilament triplet. Reconstitution of the three bovine polypeptides into 10-nm filaments was accomplished under conditions in which the NF3 protein was limiting. Reassembly of 10-nm filaments with varying amounts of NF2 and NF1 indicate that the NF3 homopolymer has a limiting capacity to bind NF2 and NF1 and is saturated at a molar ratio of 2:2:1 (NF3:NF2:NF1). Isolation of the neurofilament complex at various stages of rat brain maturation indicates that NF3 and NF2 are integrated into the neurofilament complex as early as embryonic day 17, while NF1 copurifies with these proteins at postnatal day 16, eventually reaching a molar stoichiometry of 2:2:1 in the adult rat. The molecular stoichiometry of the neurofilament proteins, the differential integration of these proteins during brain development, and the variation of the molar composition between mammalian species suggest accessory roles for the NF2 and NF1 proteins in the neurofilament complex. PMID- 2993287 TI - Rat hepatic cytosolic phosphoenolpyruvate carboxykinase (GTP). Structures of the protein, messenger RNA, and gene. AB - The primary structure of the messenger RNA coding for cytosolic phosphoenolpyruvate carboxykinase was determined by sequencing cDNA and genomic DNA and by primer extension of the mRNA. The molecule is 2624 nucleotides in length; this includes 143 nontranslated nucleotides at the 5' end and 615 nontranslated nucleotides at the 3' end. The 3' nontranslated sequence contains a 102-base pair region of alternating purine-pyrimidine nucleotides (the majority of which are UpG dinucleotides), several direct repeats and palindromic sequences, and 8 CpG dinucleotides. The corresponding segment of the phosphoenolpyruvate carboxykinase gene thus has characteristics which favor the formation of Z-DNA. The amino acid sequence of phosphoenolpyruvate carboxykinase was deduced from the mRNA sequence and confirmed by fast atom bombardment mass spectrometric analysis of peptides generated with trypsin and Staphylococcus aureus V8 protease. The protein consists of 621 amino acids and has a molecular weight of 69,289. Charon 4A lambda bacteriophage clones containing genomic DNA coding for phosphoenolpyruvate carboxykinase were isolated from a library of partial HaeIII digests of rat liver DNA. Two clones, lambda PC112 and lambda PC103, contained the entire coding region in 15-kilobase inserts and were used to subclone the gene into pBR322 as EcoRI, BamHI, or SstI-KpnI fragments. Using these subclones, the structure of the phosphoenolpyruvate carboxykinase gene was determined by S1 nuclease mapping, R-loop analysis, and DNA sequencing. The gene is composed of 10 exons and 9 introns with a total length of 6.0 kilobases. The transcription initiation site of the gene was determined by a combination of in vitro transcription in a HeLa cell lysate system, primer extension of mRNAPEPCK, and S1 nuclease mapping. In vitro transcription of purified DNA templates revealed three RNA polymerase II-dependent start sites. Two sites were separated by 600 base pairs on the coding strand and the third site was on the noncoding strand. The products of S1 nuclease mapping and primer extension from a BglII site were compared in order to determine which of the coding strand initiation sites was expressed in vivo. In both cases a 69-base pair fragment was generated and the 5' end of this corresponded to a thymidine residue identified in a sequence ladder of the genomic DNA coding strand. We conclude that mRNAPEPCK synthesis initiates with an adenine residue 69 base pairs 5' of the BglII site; this corresponds to the 3' most transcription initiation site determined in vitro. PMID- 2993288 TI - P-57 is a neural specific calmodulin-binding protein. AB - P-57 is a novel calmodulin-binding protein which has recently been isolated from bovine cerebral cortex (Andreasen, T. J., Luetje, C. W., Heideman, W., and Storm, D. R. (1983) Biochemistry 22, 4615-4618). In contrast to all other calmodulin binding proteins characterized thus far, P-57 has equivalent or higher affinity for calmodulin in the absence of free Ca2+ compared to the presence of Ca2+. In this study, the distribution of P-57 in other tissues and within brain was examined using a radioimmune assay and photoaffinity labeling with azido-125I calmodulin. P-57 was not found in tissues other than brain, retina, and spinal cord. Within brain, P-57 levels varied from 0.1% of the total protein in white matter regions to about 0.5% in cell body-rich fractions. The protein was found in both membrane and soluble fractions. P-57 is the most abundant calmodulin binding protein in brain and appears to be neural specific. The concentrations of P-57 in brain and its affinity for calmodulin in the absence of Ca2+ are sufficient to complex a significant fraction of the total calmodulin present. PMID- 2993289 TI - The voltage-sensitive Na+/H+ exchange in sea urchin spermatozoa flagellar membrane vesicles studied with an entrapped pH probe. AB - Flagellar plasma membrane vesicles were isolated from sea urchin sperm using osmotic lysis. A membrane impermeant fluorescence pH indicator, pyranine, was incorporated into the vesicles as they resealed after lysis and was used to measure the intravesicular pH (pHi). Addition of Na+ rapidly alkalinized the pHi of vesicles prepared with an internal acidic pH gradient. The pHi increase showed ionic selectivity in the order of Na+ greater than Li+ much greater than K+ approximately equal to Cs+ approximately equal to O. Complete removal of monovalent anions such as Cl- and HCO3- did not affect the exchange, thus ruling out the participation of an anion carrier in the process. The optimal operation of the exchanger, however, required the presence of a transmembrane potential, which could be generated by the diffusion potential of either K+, a naturally permeant ion, or Cs+ which was artificially made permeant by the ionophore valinomycin. Depolarization inhibited the exchange in both the forward and the reverse directions, which is consistent with the voltage-gated electroneutral exchange mechanism proposed previously for this exchanger (Lee, H. C. J. Biol. Chem. (1984) 259, 15315-15319). The voltage sensitivity of the Na+/H+ exchanger was found to be modulated by the presence of Mg2+. A model involving the screening of the internal surface potential was proposed to account for the Mg2+ effect. The vesicle preparation used in this study allows complete control of the internal contents and represents a major simplification of the system as compared with the intact sperm and the isolated flagella used previously. PMID- 2993290 TI - The A1 adenosine receptor. Identification of the binding subunit by photoaffinity cross-linking. AB - Adenosine modifies the catalytic activity of adenylate cyclase through both inhibitory (A1 or Ri) as well as stimulatory (A2 or Ra) cell surface receptors. We developed 125I-labeled N6-2-(4-aminophenyl)ethyladenosine as a selective ligand to probe the structure of A1 receptors. The binding of this radioligand to rat cerebral cortex or adipocyte membranes is saturable, reversible, and of high affinity (KD approximately 2 nM). A1 receptor agonists antagonize binding stereoselectivity and with a potency order appropriate for A1 receptors. The heterobifunctional cross-linking reagent N-succinimidyl-6-(4-azido-2 nitrophenylamino)hexanoate covalently couples the radioligand to a protein of Mr = 38,000 in both tissues as demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Inhibition of covalent labeling by adenosine analogs exhibited the stereoselectivity and potency order typical of A1 receptor ligands. Guanine nucleotides reduced both specific binding and covalent incorporation of the radioligand, evidence that the radioligand is an A1 receptor agonist. These results suggest that the A1 receptor binding subunit of both brain and adipocytes resides on a protein of Mr = 38,000. The new radioligand should prove useful in studying the structure and regulation of A1 receptors. PMID- 2993291 TI - Hepatic adenylate cyclase. Development-dependent coupling to the beta-adrenergic receptor in the neonate. AB - Guanine nucleotide-dependent modulation of agonist binding to the beta-receptor reflects coupling of the receptor to the nucleotide regulatory protein. Similarly, guanine nucleotide-dependent stimulation of adenylate cyclase can be used as an index of coupling between the regulatory protein and the catalytic unit of the cyclase. Using both approaches we have studied coupling in the beta adrenergic receptor-adenylate cyclase system in rabbit liver during neonatal development. With [3H]dihydroalprenolol as ligand, the Bmax was relatively unchanged (200-300 fmol/mg of protein) between birth and end of day 1 and was similar to adult values. Guanyl-5'-yl imidodiphosphate-dependent shift in agonist (l-isoproterenol) competition curves was biphasic, decreasing from 10-fold in membranes isolated from animals at term to about 6-fold in membranes from 6-h-old neonates, and increasing progressively in older animals to a maximal measurable value of 42-fold in the adult. The ability of guanyl-5'-yl imidodiphosphate, GTP, GTP plus isoproterenol, NaF, or forskolin to activate adenylate cyclase was also biphasic and age-dependent. With Mn2+ the measured activity was not at any time greater than the activity at term. Pretreatment of membranes with cholera toxin resulted in differential levels of enhancement of adenylate cyclase activity wherein much lower enhancement was observed in membranes from neonatal animals. With [32P]NAD as substrate, cholera toxin-catalyzed ADP-ribosylation of membranes indicated development-dependent accumulation of Ns peptides. From these results we suggest that there is a decreased efficiency in the coupling of the beta adrenergic receptor to hepatic adenylate cyclase in early neonatal life. The molecular basis for the biphasic nature of the coupling is presently unclear. PMID- 2993292 TI - Covalent labeling of opioid receptors with radioiodinated human beta-endorphin. Identification of binding site subunit. AB - 125I-beta-Endorphin (human) binds with high affinity, specificity, and saturability to rat brain and neuroblastoma X glioma hybrid cell (NG 108-15) membranes. Dissociation constants and binding capacities were obtained from Scatchard plots and are 2 nM and 0.62 pmol/mg of protein for rat whole brain and 6 nM and 0.8 pmol/mg of protein for NG 108-15 cells. Results from competition experiments also indicate that this ligand interacts with high affinity with both mu and delta opioid binding sites, with a slight preference for mu sites, while exhibiting low affinity at kappa sites. We have demonstrated that human 125I-beta endorphin is a useful probe for the investigation of the subunit structure of opioid receptors. The specific cross-linking of this ligand has revealed the presence of four reproducible bands or areas after sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography at 65, 53, 38, and 25 kDa. All labeled bands seem to be opioid receptor related since they are eliminated when binding is carried out in an excess of various opiates. The evidence we have obtained using rat whole brain (delta congruent to mu), rat thalamus (largely mu), bovine frontal cortex (delta:mu congruent to 2:1), and NG 108-15 cells (delta) demonstrates that different labeling patterns are obtained when mu and delta binding sites are cross-linked. The pattern obtained on sodium dodecyl sulfate-polyacrylamide gel electrophoresis from cross-linked mu sites contains a major (heavily labeled) component of 65 kDa and a minor component of 38 kDa, while patterns from delta sites contain a major labeled component of 53 kDa. This 53-kDa band appears clearly in extracts from NG 108-15 cells and bovine frontal cortex, while in rat whole brain a diffusely labeled region is present between 55 and 41 kDa. In addition, NG 108-15 cells also display a minor labeled component at 25 kDa. The relationship of the minor bands to the major bands is not clear. PMID- 2993293 TI - Identification of a receptor for atrial natriuretic factor in rabbit aorta membranes by affinity cross-linking. AB - Binding sites in rabbit aorta membranes for atrial natriuretic factor (ANF) have been specifically and covalently labeled by two methods. In the first, the photoreactive analog of ANF, 125I-azidobenzoyl-ANF, was synthesized and used to photoaffinity label ANF receptors. In the second, 125I-ANF was covalently attached to its binding site by treatment of the 125I-ANF-receptor complex with bifunctional cross-linking agents. Analysis of the labeled proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed that by both methods the same three protein bands were labeled. These bands had apparent molecular masses of 60,000, 70,000, and 120,000 daltons. With the photoaffinity label, half-maximal inhibition of labeling of each of these bands was achieved when approximately 200 pM of unlabeled ANF was included in the binding assay. These results suggest that these three different polypeptides are specific components of ANF receptors in rabbit aorta membranes. PMID- 2993294 TI - Electron paramagnetic resonance studies of mammalian succinate dehydrogenase. Detection of the tetranuclear cluster S2. AB - Electron paramagnetic resonance studies of Complex II from the mitochondrial respiratory chain and soluble preparations of succinate dehydrogenase have, for the first time, identified a signal arising from a [4Fe-4S]1+ cluster, S2, in dithionite-reduced samples. Redox titrations, monitored by electron paramagnetic resonance spectroscopy demonstrate that this signal appears at the same midpoint potential as the enhancement of the spin relaxation properties of the [2Fe-2S]1+ center, S1, in both Complex II and reconstitutively active soluble enzyme. The results complement recent magnetic circular dichroism studies of succinate dehydrogenase (Johnson, M. K., Morningstar, J. E., Bennett, D. E., Ackrell, B. A. C., and Kearney, E. B. (1985) J. Biol. Chem. 260, 7368-7378) which assigned cluster S2 as a [4Fe-4S]2+,1+ center and provide evidence for spin interaction between the paramagnetic reduced forms of centers S1 and S2. PMID- 2993295 TI - Complete nucleotide and derived amino acid sequences of the fourth component of mouse complement (C4). Evolutionary aspects. AB - The nucleotide sequence coding for the fourth component of mouse complement (C4) has been determined from a cloned genomic DNA fragment and a cloned cDNA fragment. The amino acid sequence of the protein was deduced. The single chain precursor protein (pro-C4) consists of 1719 amino acid residues. The mature beta, alpha, and gamma subunits contain 654, 766, and 291 amino acids, respectively. One potential carbohydrate attachment site is predicted for the beta chain, three for the alpha chain, and none for the gamma chain. From a comparison with human C4 cDNA sequence an extensive overall sequence homology, 79% in nucleotides and 76% in amino acids, is observed. There is conservation in both the position and number of cysteine residues in human and mouse C4. We compared the mouse C4 amino acid sequences with those of mouse C3 and human alpha 2-macroglobulin and the evolutionary relationship among these three proteins is discussed. PMID- 2993296 TI - An alternate pathway for the processing of the prolipoprotein signal peptide in Escherichia coli. AB - Previous studies showed that when the signal sequence plus 9 amino acid residues from the amino terminus of the major lipoprotein of Escherichia coli was fused to beta-lactamase, the resulting hybrid protein was modified, proteolytically processed, and assembled into the outer membrane as was the wild-type lipoprotein (Ghrayeb, J., and Inouye, M. (1983) J. Biol. Chem. 259, 463-467). We have constructed several hybrid proteins with mutations at the cleavage site of the prolipoprotein signal peptide. These mutations are known to block the lipid modification of the lipoprotein at the cysteine residue, resulting in the accumulation of unprocessed, unmodified prolipoprotein in the outer membrane. The mutations blocked the lipid modification of the hybrid protein. However, in contrast to the mutant lipoproteins, the cleavage of the signal peptides for the mutant hybrid proteins did occur, although less efficiently than the unaltered prolipo-beta-lactamase. The mutant prolipo-beta-lactamase proteins were cleaved at a site 5 amino acid residues downstream of the prolipoprotein signal peptide cleavage site. This new cleavage between alanine and lysine residues was resistant to globomycin, a specific inhibitor for signal peptidase II. This indicates that signal peptidase II, the signal peptidase which cleaves the unaltered prolipo-beta-lactamase, is not responsible for the new cleavage. The results demonstrate that the cleavage of the signal peptide is a flexible process that can occur by an alternative pathway when the normal processing pathway is blocked. PMID- 2993297 TI - Leukotriene A3. A poor substrate but a potent inhibitor of rat and human neutrophil leukotriene A4 hydrolase. AB - Analysis of leukotriene B4 production by purified rat and human neutrophil leukotriene (LT) A4 hydrolases in the presence of 5(S)-trans-5,6-oxido-7,9-trans 11-cis-eicosatrienoic acid (leukotriene A3) demonstrated that this epoxide is a potent inhibitor of LTA4 hydrolase. Insignificant amounts of 5(S), 12(R) dihydroxy-6-cis-8,10-trans-eicosatrienoic acid (leukotriene B3) were formed by incubation of rat neutrophils with leukotriene A3 or by the purified rat and human LTA4 hydrolases incubated with leukotriene A3. Leukotriene A3 was shown to be a potent inhibitor of leukotriene B4 production by rat neutrophils and also by purified rat and human LTA4 hydrolases. Covalent coupling of [3H]leukotriene A4 to both rat and human neutrophil LTA4 hydrolases was shown, and this coupling was inhibited by preincubation of the enzymes with leukotriene A4. Preincubation of rat neutrophils with leukotriene A3 also prevented labeling of LTA4 hydrolase by [3H]leukotriene A4. This result indicates that leukotriene A3 prevents covalent coupling of the substrate leukotriene A4 and inhibits the production of leukotriene B4 by blocking the binding of leukotriene A4 to the enzyme. PMID- 2993298 TI - Regulation of calcium transport by protein phosphatase activity associated with cardiac sarcoplasmic reticulum. AB - Canine cardiac sarcoplasmic reticulum is phosphorylated by an endogenous calcium calmodulin-dependent protein kinase on a 22,000 proteolipid, called phospholamban. Phosphorylation by the calcium-calmodulin-dependent protein kinase is associated with stimulation of the initial rates of calcium transport (Davis, B. A., Schwartz, A., Samaha, F. J., and Kranias, E. G. (1983) J. Biol. Chem. 258, 13587-13591). The present study shows that protein phosphatase activity, associated with canine cardiac sarcoplasmic reticulum vesicles, can catalyze dephosphorylation of the calcium-calmodulin-dependent sites on phospholamban. The activity was maximally stimulated by manganese; fluoride was inhibitory, but its effect was reversible. Dephosphorylation of phospholamban, which was prephosphorylated by calcium-calmodulin-dependent protein kinase, resulted in a reduction of the stimulation on calcium transport rates, particularly at submaximal calcium concentrations. The decrease in calcium transport was associated with a statistically significant decrease in the apparent affinity (EC50) for calcium. Rephosphorylation of phospholamban by the endogenous calcium calmodulin-dependent protein kinase caused full recovery of the stimulation on calcium transport rates and reversal of the effects mediated by the protein phosphatase. Thus, the calcium pump in cardiac sarcoplasmic reticulum appears to be under reversible regulation mediated by endogenous calcium-calmodulin dependent protein kinase and protein phosphatase. Such regulation may represent an important control mechanism for the myocardium. PMID- 2993299 TI - Insulin-stimulated phosphoinositide metabolism in isolated fat cells. AB - Treatment of isolated fat cells with insulin produced increases of up to 4.8-fold in the incorporation of [3H]inositol into phosphatidylinositol. This effect of insulin was both time- and dose-dependent with half-maximal stimulation at 30 microunits/ml of insulin. Insulin increased the labeling of phosphatidylinositol and phosphatidylinositol 4,5-bisphosphate but not phosphatidylinositol 4 monophosphate in cells which had been preincubated with [3H]inositol for 90 min. Incubation of the cells in a Ca2+-free buffer increased the basal level of phosphatidylinositol labeling and enhanced the effect of insulin. Glucagon and isoprenaline, both of which stimulate lipolysis, had no effect on phosphatidylinositol labeling but did potentiate insulin-stimulated incorporation of [3H]inositol into phosphatidylinositol. Phosphoinositide breakdown was measured by the accumulation of inositol phosphates. Insulin did not increase the level of the inositol phosphates at all concentrations of the hormone tested. By comparison, phenylephrine and vasopressin were able to stimulate phosphoinositide breakdown. Pretreatment of the cells with insulin enhanced the effect of phenylephrine on inositol phosphates' accumulation, suggesting that insulin may potentiate phenylephrine-mediated phosphoinositide turnover. From these data we conclude that insulin stimulates the de novo synthesis of phosphatidylinositol and phosphatidylinositol 4,5-biphosphate, but has no effect on phosphoinositide breakdown. PMID- 2993300 TI - Bovine brain calmodulin-dependent protein phosphatase. Regulation of subunit A activity by calmodulin and subunit B. AB - Calmodulin-dependent protein phosphatase isolated from bovine brain consists of a catalytic subunit A (Mr = 60,000) and a regulatory subunit B (Mr = 19,000) present in equal molar ratios. The two subunits were dissociated by gel filtration in 6 M urea and reconstituted to investigate the role of calmodulin and subunit B in regulating the phosphatase activity of subunit A. The activity of subunit A was stimulated 2-fold by calmodulin, 13-fold by subunit B, and 21 fold by both, indicating that the effects of both were synergistic. Maximum stimulation by calmodulin was observed at a calmodulin to subunit A molar ratio of 2:1 in the presence or absence of subunit B, whereas that by subunit B was observed at a B to A molar ratio of 3:1 in the presence or absence of calmodulin. Calmodulin and subunit B increased the Vmax of subunit A 2- and 5-fold, respectively, but had little effect on the Km for casein. The specific activity of the phosphatase reconstituted from subunits A and B reached 86% that of the native enzyme, whereas that of the holoenzyme reached 90%. Subunit B, even though similar to calmodulin in many respects, did not stimulate the activity of native phosphatase, suggesting that it cannot substitute for calmodulin. Limited trypsinization of subunit A increased its catalytic activity to the level observed with calmodulin; and this activity was further stimulated by subunit B but not by calmodulin. These results indicate that subunit A of phosphatase contains one catalytic domain and two distinct regulatory domains, one for calmodulin, and another for subunit B, that these two proteins do not substitute for one another and that they stimulate subunit A synergistically. PMID- 2993301 TI - Differential RNA splicing and post-translational cleavages in the human salivary proline-rich protein gene system. AB - The nucleotide sequences of cDNAs coding for human salivary proline-rich proteins (PRPs) were determined. Clones cP1 and cP2 contain repetitive regions in which sites for the restriction enzyme HaeIII occur repeatedly; they code for the precursors of acidic PRPs. Clones cP3 to cP7 contain repetitive regions in which BstNI sites occur repeatedly; they code for precursors of basic and glycosylated PRPs. The clones cP3, cP4, and cP5 are identical except that cP4 and cP5 are missing 399 and 459 base pairs, respectively, from the repetitive region of cP3. The sequences at these deletion end points are homologous to the consensus sequences of RNA splicing donor and acceptor sites. This strongly suggests that all three cDNAs are derived from the transcript of a single gene via differential RNA splicing. All of the precursor proteins share a feature--the N-terminal region, following the signal peptide, is acidic, while the remainder of the molecule, made of proline-rich repeats of about 21 amino acids, is basic. Each precursor can generate multiple PRPs by various post-translational cleavages on the carboxylic side of specific arginine residues. The data show how differential RNA splicing and post-translational cleavages could generate a large number of proteins, such as those found in saliva, from a much smaller number of genes. PMID- 2993302 TI - A single cardiac troponin T gene generates embryonic and adult isoforms via developmentally regulated alternate splicing. AB - A single cardiac troponin T gene generates two mRNA products by developmentally regulated alternate splicing. Nucleotide sequence of the entire 18 exon gene and both representative cDNAs demonstrate that the two mRNAs differ by the presence or absence of a single internal coding exon. Both mRNA products appear to be generated from a single primary transcript; however, one mRNA splice product predominates in early embryonic cardiac muscle while the other vastly predominates in adult cardiac muscle. The corresponding embryonic and adult cardiac troponin T proteins differ by the inclusion or exclusion, respectively, of an internal, highly acidic 10 amino acids near the amino terminus. Unusual features of the variable peptide region and its restriction to embryonic stages suggest that it might play a specialized role during sarcomere assembly in embryonic striated muscle. In addition to developmental regulation of RNA processing, the cardiac troponin T gene also demonstrates complex tissue-specific expression. We have previously shown that this single cardiac troponin gene is regulated according to two different and tissue-specific regulatory programs. Here we demonstrate that a single promoter is utilized for both expression patterns. The cardiac troponin T gene also has several interesting structural features including a pseudoexon and an exon only six nucleotides in length. The size of this exon establishes a new lower limit for the number of nucleotides that can be recognized as an exonic sequence. PMID- 2993303 TI - CL glycoprotein is the tissue form of type VI collagen. AB - CL glycoprotein (CLGP), the 140,000-dalton collagenous glycoprotein, has been isolated from fetal bovine aorta and nuchal ligament, in milligram amounts in its reduced and alkylated form, using a multistage procedure. This material exhibited a characteristic amino acid composition with a consistent ratio of hydroxylysine to hydroxyproline (approximately 1:1). Digestion of CLGP with bacterial collagenase yielded three discrete noncollagenous fragments. Monospecific anti CLGP antiserum exhibited strong cross-reactivity with the pepsin-resistant polypeptides of type VI collagen. CLGP was also prepared in the unreduced disulfide-bonded form and in a partially reduced form, using brief treatment with cysteine. On treatment with pepsin these preparations yielded resistant peptides corresponding in size to the longer and shorter forms, respectively, of type VI collagen. A slightly larger, soluble form of CLGP (Mr = 150,000) was detected in the media from cultures of aortic smooth muscle cells and nuchal ligament fibroblasts. The evidence indicates that CLGP is the native form in which type VI collagen is present in the tissues and that it consists of three structurally distinct polypeptide chains, each about 140 kDa in mass, which are disulfide bonded into a triple-helical molecule. The CLGP molecules appear to be present in the tissues as dimers and larger aggregates, stabilized by intermolecular disulfide bonding. The distribution of type VI collagen will thus be as described in our earlier immunofluorescence studies with anti-CLGP antiserum (Gibson, M.A., and Cleary, E.G. (1983) Collagen Relat. Res. 3, 469-488). PMID- 2993304 TI - Mossbauer, EPR, and magnetization studies of the Azotobacter vinelandii Fe protein. Evidence for a [4Fe-4S]1+ cluster with spin S = 3/2. AB - We have studied the Fe protein (Av2) of the Azotobacter vinelandii nitrogenase system with Mossbauer and EPR spectroscopies and magnetic susceptometry. In the oxidized state the protein exhibits Mossbauer spectra typical of diamagnetic [4Fe 4S]2+ clusters. Addition of Mg.ATP or Mg.ADP causes a pronounced decline in the quadrupole splitting of the Mossbauer spectra of the oxidized protein. Our studies show that reduced Av2 in the native state is heterogeneous. Approximately half of the molecules contain a [4Fe-4S]1+ cluster with electronic spin S = 1/2 and half contain a [4Fe-4S]1+ cluster with spin S = 3/2. The former yields the characteristic g = 1.94 EPR signal whereas the latter exhibits signals around g = 5. The magnetization of reduced Av2 is dominated by the spin S = 3/2 form of its [4Fe-4S]1+ clusters. These results explain a long standing puzzle, namely why the integrated spin intensity of the g = 1.94 EPR signal is substantially less than 1 spin/4 Fe atoms. In 50% ethylene glycol, 90% of the clusters are in the spin S = 1/2 form whereas, in 0.4 M urea, 85% are in the S = 3/2 form. In 0.4 M urea, the EPR spectrum of reduced Av2 exhibits well defined resonances at g = 5.8 and 5.15, which we assign to the S = 3/2 system. The EPR and Mossbauer studies yield a zero field splitting of 2D approximately equal to -5 cm-1 for this S = 3/2 state. PMID- 2993305 TI - Morphological transformation of Chinese hamster cells by acylpeptides, inhibitors of cAMP phosphodiesterase, produced by Bacillus subtilis. AB - Treatment of Chinese hamster ovary cells in vitro with acylpeptide, an inhibitor of adenosine 3':5'-cyclic monophosphate (cAMP) phosphodiesterase, produced by Bacillus subtilis C-756 converts the culture from one of randomly oriented, epithelial-like cells to one of elongated, fibroblast-like cells. This transformation is similar to the effect of N6,O2'-dibutyryl adenosine 3':5' cyclic monophosphate (dibutyryl cAMP), but is manifested at a lower concentration of the inhibitor than with dibutyryl cAMP. The intracellular level of cAMP in the cells grown with the inhibitor is 3-4-fold higher than the level without the inhibitor. The transformation is reversible and, when reversed, the level of cAMP simultaneously decreases to the level without the inhibitor. The acylpeptide has cytolytic activity as well as inhibitory activity and the former activity is prevented by the addition of serum. PMID- 2993306 TI - Phospholipid organization in H,K-ATPase-containing membranes from pig gastric mucosa. AB - The transverse distribution of the phospholipids in vesicular H+-translocating membranes prepared from pig gastric mucosa was investigated with the aid of phospholipase C, sphingomyelinase, and trinitrobenzenesulfonic acid. The major part (80-90%) of the phosphatidylcholine and the phosphatidylethanolamine, 60% of the phosphatidylserine, and 45% of the sphingomyelin was located on the external, cytoplasmic side of the vesicle membranes. After treatment with phospholipase C the vesicles still behaved as osmometers and appeared as closed vesicles on the electron micrographs. 31P NMR indicated that the phospholipids in untreated vesicles as well as the unhydrolyzed phospholipids in phospholipase C-treated vesicles were arranged in lamellar structures. The 31P NMR spectrum of untreated vesicles to which Pr3+ ions had been added supported the conclusion that the major part of the membrane phospholipids was located on the external surface of the vesicles. A small fraction of the lipids, 3.6 mol %, was found to consist of glycosphingolipids which occurred at a concentration of 52 nmol/mg of protein. PMID- 2993307 TI - A heterozygous collagen defect in a variant of the Ehlers-Danlos syndrome type VII. Evidence for a deleted amino-telopeptide domain in the pro-alpha 2(I) chain. AB - A structural defect in the alpha 2(I) chain of type I collagen was characterized in a new case of the Ehlers-Danlos syndrome type VII. The patient's skin, fascia, and bone collagens all showed an abnormal additional chain, pN-alpha 2(I)s, running slower than the alpha 2(I) chain on electrophoresis. The extension was shown to be on the amino-terminal fragment of pN-alpha (I)s by cleavage with human collagenase, but pepsin was unable to convert pN-alpha 2(I)s to alpha 2(I). Skin collagen was 4-fold more extractable and contained fewer beta-dimers and a lower concentration of cross-linking amino acids than control skin collagen. Electron micrographs of both dermis and bone showed markedly irregular ragged outlines of the collagen fibrils in cross-section, although the patient had no clinical signs of bone disease. Procollagen secreted by her skin fibroblasts in culture showed equal amounts of the normal and abnormal alpha 2(I) chains on pepsin digestion. Before pepsin, the pN-alpha 2(I) component ran as a doublet on electrophoresis; pepsin removed only the normal slower chain. The suspected deletion in pN-alpha 2(I)s was traced by CNBr peptide analysis to the N propeptide fragment, which behaved on electrophoresis about 15-20 residues smaller than that from the normal pN-alpha 2(I) chain. The simplest genetic explanation is a spontaneous heterozygote in which one normal and one abnormal allele for the pro-alpha 2(I) gene are expressed, the protein defect being a deletion of the junction domain that spans the N-propeptidase cleavage site and the N-telopeptide cross-linking sequence. PMID- 2993308 TI - Biosynthesis and processing of the type II insulin-like growth factor receptor in H-35 hepatoma cells. AB - The biosynthesis and post-translational processing of the insulin-like growth factor II (IGF-II) receptor has been studied in H-35 hepatoma cells using a specific polyclonal anti-receptor immunoglobulin preparation. Cells were pulse labeled with [35S]methionine followed by incubation with excess unlabeled methionine (chase). Gel electrophoresis of the immunoadsorbed receptors shows that the receptor is first synthesized as a 245-kDa precursor which is transformed to the mature 250-kDa form with a half-time of about 2 h. The 245-kDa precursor could also be labeled biosynthetically with [3H]mannose, only one-half of which was ultimately found associated with the 250-kDa product. Neuraminidase converts the 250-kDa receptor species to a 245-kDa form. Whereas the 250-kDa receptor is insensitive to detectable cleavage by endoglycosidase H, digestion of the 245-kDa species with this enzyme produces a 232-kDa form. A similar 232-kDa receptor species accumulates in H-35 cells incubated with tunicamycin (2 micrograms/ml). This tunicamycin-induced aglyco-receptor is not further processed to the 250-kDa form. Monensin (50 nM) blocks receptor processing at the 245-kDa stage. Endoglycosidase H treatment of the monensin-induced 245-kDa species indicates that this is a mixture of partially processed precursors having equivalent Mr. No evidence was obtained for the presence of O-linked oligosaccharides on the IGF-II receptor. The IGF-II binding activity of the three different biosynthetic forms of the receptor was assessed by affinity cross linking of 125I-IGF-II to the receptors using disuccinimidyl suberate. Both the mature 250-kDa receptor and the neuraminidase-digested 245-kDa form specifically bound 125-I-IGF-II. However, the 232-kDa aglyco-receptor had no detectable IGF-II binding activity using this method. In summary, these studies show: 1) that the H 35 cell IGF-II receptor is synthesized first as a 245-kDa precursor having 4-6 high-mannose oligosaccharide side chains, 2) processing of the receptor oligosaccharides by mannose removal and terminal sialylation converts the 245-kDa precursor to the 250-kDa mature product which has been previously identified as the functional receptor in the plasma membrane, 3) the apparent molecular mass of the receptor in the absence of N-glycosylation is 232-kDa, and 4) glycosylation of the IGF-II receptor is required for the acquisition of IGF-II binding activity. PMID- 2993309 TI - Factors influencing changes in moisture content during storage of freeze-dried vaccines in vials. AB - Previous studies have demonstrated that the water content of freeze-dried vaccines increases during storage. The reasons for these variations in water content are discussed in this paper. Different possible mechanisms have been considered: microleakages at the closure sealing point, water vapour transfer through the closure (permeation, loss and uptake of water by the stopper). Several types of vials and closures were studied, under different conditions of storage. Regardless of the vial form selected, the probability of microleakages was low. The stopper would seem to play an important role in the water transfer mechanisms. The moisture content of the stopper determines its equilibrium relative humidity (ERH) and whether there will be moisture transfer between the stopper and the vial contents and between the stopper and the storage atmosphere. PMID- 2993310 TI - Quantitation of glucagon by radioreceptorassay. AB - Receptors for glucagon on rat liver membranes were characterized. They bound [125I] glucagon rapidly in a specific and saturable way. Addition of unlabelled glucagon displaced [125I] glucagon from the binding sites in a concentration dependent way. Concentrations from 10(-9) to 10(-8) M of glucagon caused a linear reduction of binding of labelled glucagon. This concentration interval was used for a three-point assay which fulfilled statistical requirements for validity. Individual assays normally resulted in potency estimates of high precision and statistical weight. Mean values for glucagon activity of preparations tested by receptor assay were within the fiducial limits (P = 0.95) for corresponding activity determined by the rabbit blood glucose method. The receptor assay is less time consuming and requires only part of one rat liver while the in vivo assay uses 16 rabbits. Thus, the receptor assay is less resource demanding and should serve well as a screening instrument for control of potency of glucagon preparations. PMID- 2993311 TI - Establishment of a statistical base for use of ELISA in diagnostic serology for infectious bovine rhinotracheitis. AB - The critical statistical parameters of an enzyme-linked immunosorbent assay were determined to enable quantitation of antibody responses in cattle affected with infectious bovine rhinotracheitis. A system of controlling well-to-well variations in optical density reading across a microtitre plate was evolved and dose--response assays were carried out to determine the dilution of serum which gave the greatest discrimination between acute and convalescent sera from an infected animal. Use of a standard serum was studied in further assays. An increase in optical density value of 0.15 was set as a diagnostic criterion for a significantly rising antibody response. This compared well with the conventional criterion of a fourfold rise in virus neutralizing antibody titre. PMID- 2993312 TI - Release of superoxide and change in morphology by neutrophils in response to phorbol esters: antagonism by inhibitors of calcium-binding proteins. AB - The ability of phorbol derivatives to function as stimulating agents for superoxide (O2-) release by guinea pig neutrophils has been evaluated and compared to the known ability of each compound to activate protein kinase C. Those that activate the kinase also stimulate O2- release, while those that are inactive with respect to the kinase have no effect on O2- release. The same correlation was observed with respect to the ability of phorbol esters to induce morphological changes in neutrophils, i.e., vesiculation and reduction in granule content. Certain phenothiazines and naphthalene sulfonamides that are known antagonists of calcium-binding proteins blocked both phorbol ester-induced O2- release and morphological changes in these cells. PMID- 2993314 TI - Selective destruction of nerve growth factor receptor-bearing cells in vitro using a hybrid toxin composed of ricin A chain and a monoclonal antibody against the nerve growth factor receptor. AB - A hybrid toxin composed of ricin A chain and a monoclonal antibody directed against the rat nerve growth factor (NGF) receptor (192-IgG) was prepared using the heterobifunctional cross-linking agent N-succinimidyl-3-(2-pyridyldithio) propionate and purified by affinity chromatography. Characterization studies showed that the hybrid, 192-s-s-A, displaced bound 125I-labeled 192-IgG from rat superior cervical ganglion (SCG) membranes with an IC50 3-5 times lower than that of unconjugated 192-IgG. When incubated with cultured rat SCG neurons, 192-s-s-A inhibited protein synthesis in a concentration-dependent fashion. The effect of 192-s-s-A on these neurons was reversed by coincubation with an excess of 192 IgG. The IC50 of 192-s-s-A on protein synthesis in rat SCG neurons was 4 nM. Intact ricin and ricin A chain inhibited protein synthesis in these neurons with IC50 values of 5 pM and 500 nM, respectively. The 192-s-s-A hybrid had no effect on mouse SCG neurons or a human melanoma cell line known to have NGF receptors. This is consistent with the finding that 192-IgG recognizes only the rat NGF receptor. Also, 192-s-s-A did not inhibit protein synthesis in primary cultures of rat skeletal muscle or Vero cells, which do not have cell surface receptors for NGF. 192-s-s-A was able to inhibit protein synthesis in PC12 cells but the potency was 10-100 times less in these cells compared to rat SCG neurons. Ricin and A chain were also 10-100 times less potent in PC12 cells than neurons. Rat SCG neurons exposed to 192-s-s-A lost their refractile appearance under phase contrast optics, showed granular degeneration of neurites, and died. Thus the decreased protein synthesis caused by the hybrid toxin correlated with the morphological destruction of the neurons. 192-s-s-A represents a potentially powerful tool by which to selectively destroy NGF receptor-bearing cells in vitro. The hybrid toxin may prove useful as an in vivo toxin. PMID- 2993313 TI - Isolation of an adhesion-mediating protein from chick neural retina adherons. AB - Adherons are high molecular weight glycoprotein complexes which are released into the growth medium of cultured cells. They mediate the adhesive interactions of many cell types, including those of embryonic chick neural retina. The cell surface receptor for chick neural retina adherons has been purified, and shown to be a heparan sulfate proteoglycan (Schubert, D., and M. LaCorbiere, 1985, J. Cell Biol., 100:56-63). This paper describes the isolation and characterization of a protein in neural retina adherons which interacts specifically with the cell surface receptor. The 20,000-mol-wt protein, called retinal purpurin (RP), stimulates neural retina cell-substratum adhesion and prolongs the survival of neural retina cells in culture. The RP protein interacts with heparin and heparan sulfate, but not with other glycosaminoglycans. Monovalent antibodies against RP inhibit RP-cell adhesion as well as adheron-cell interactions. The RP protein is found in neural retina, but not in other tissues such as brain and muscle. These data suggest that RP plays a role in both the survival and adhesive interactions of neural retina cells. PMID- 2993315 TI - Intracellular regulators of neuronal sprouting: calmodulin-binding proteins of nerve growth cones. AB - The focus of this study is a quantitative biochemical analysis of the calcium dependent interactions of calmodulin with a nerve growth cone preparation from fetal rat brain (Pfenninger, K. H., L. Ellis, M. P. Johnson, L. B. Freidman, and S. Somlo, 1983, Cell 35:573-584). The presence of calmodulin as an integral component of this preparation is demonstrated, and quantitative binding studies are presented. The binding of 125I-calmodulin to nerve growth cone material is shown to be highly specific, calcium dependent, and saturable at nanomolar calmodulin concentrations. Additionally, the growth cones' binding components appear to be membrane proteins. The individual molecular mass species of growth cone proteins displaying calcium-dependent calmodulin binding are also detailed and presented in comparison with those of synaptosomes. This analysis reveals differences between the calmodulin binding proteins of the growth cone preparation and the synaptosome fraction, suggesting the presence in growth cones of a specialized set of components which may be involved in regulatory mechanisms controlling neuritic sprouting. PMID- 2993316 TI - Intracellular pH in Dictyostelium discoideum: a 31P nuclear magnetic resonance study. AB - We have used phosphorus-31 nuclear magnetic resonance to determine intracellular pH in the cellular slime mold Dictyostelium discoideum. We devised an air-lift circulator to maintain the dense cell suspensions in a well-oxygenated and well stirred state while causing minimal perturbation to the sample flowing through the detector coils. Cells continued to develop normally in this set-up. Spectra acquired under these conditions typically show two peaks in the inorganic phosphate region corresponding to pH values of 7.16 +/- 0.03 and 6.48 +/- 0.02. These peaks are believed to represent the mitochondrial and cytosolic compartments respectively, based on a comparison of these values with published data and the collapse of the two compartments upon addition of the mitochondrial uncoupler carbonyl cyanide 4-(trifluoromethoxy)-phenylhydrazone. Dictyostelium cells show a remarkable degree of intracellular pH homeostasis. Both mitochondrial and cytosolic pH remained unchanged as extracellular pH was varied from 4.3 to 8.1. There was also no apparent change in the pH of either compartment after up to 13.5 hours' development in suspension. PMID- 2993317 TI - Electron microscopic evidence for externalization of the transferrin receptor in vesicular form in sheep reticulocytes. AB - Using ferritin-labeled protein A and colloidal gold-labeled anti-rabbit IgG, the fate of the sheep transferrin receptor has been followed microscopically during reticulocyte maturation in vitro. After a few minutes of incubation at 37 degrees C, the receptor is found on the cell surface or in simple vesicles of 100-200 nm, in which the receptor appears to line the limiting membrane of the vesicles. With time (60 min or longer), large multivesicular elements (MVEs) appear whose diameter may reach 1-1.5 micron. Inside these large MVEs are round bodies of approximately 50-nm diam that bear the receptor at their external surfaces. The limiting membrane of the large MVEs is relatively free from receptor. When the large MVEs fuse with the plasma membrane, their contents, the 50-nm bodies, are released into the medium. The 50-nm bodies appear to arise by budding from the limiting membrane of the intracellular vesicles. Removal of surface receptor with pronase does not prevent exocytosis of internalized receptor. It is proposed that the exocytosis of the approximately 50-nm bodies represents the mechanism by which the transferrin receptor is shed during reticulocyte maturation. PMID- 2993320 TI - Matrix influence on the tumor cell stimulation of fibroblast collagenase production. AB - Cocultures of rabbit fibroblasts and mouse B-16 melanoma cells produce increased levels of collagenase against type I collagen. This stimulatory effect was also found when fibroblasts were cultured in conditioned media from tumor cells. However, the level of the stimulatory factor in conditioned media was influenced by matrix deposited by fibroblasts. Thus, conditioned media collected from monolayers of B-16 plated on fibroblast matrix consistently showed high levels of the factor activity. The influence of the matrix on the level of the factor was not removed by treating the fibroblast matrix with collagenase or chondroitinase ABC and was not reproduced by collagen-coated dishes. PMID- 2993318 TI - Localization of nuclear subunits of cyclic AMP-dependent protein kinase by the immunocolloidal gold method. AB - An immunocolloidal gold electron microscopy method is described allowing the ultrastructural localization and quantitation of the regulatory subunits RI and RII and the catalytic subunit C of cAMP-dependent protein kinase. Using a postembedding indirect immunogold labeling procedure that employs specific antisera, the catalytic and regulatory subunits were localized in electron-dense regions of the nucleus and in cytoplasmic areas with a minimum of nonspecific staining. Antigenic domains were localized in regions of the heterochromatin, nucleolus, interchromatin granules, and in the endoplasmic reticulum of different cell types, such as rat hepatocytes, ovarian granulosa cells, and spermatogonia, as well as cultured H4IIE hepatoma cells. Morphometric quantitation of the relative staining density of nuclear antigens indicated a marked modulation of the number of subunits per unit area under various physiologic conditions. For instance, following partial hepatectomy in rats, the staining density of the nuclear RI and C subunits was markedly increased 16 h after surgery. Glucagon treatment of rats increased the staining density of only the nuclear catalytic subunit. Dibutyryl cAMP treatment of H4IIE hepatoma cells led to a marked increase in the nuclear staining density of all three subunits of cAMP-dependent protein kinase. These studies demonstrate that specific antisera against cAMP dependent protein kinase subunits may be used in combination with immunogold electron microscopy to identify the ultrastructural location of the subunits and to provide a semi-quantitative estimate of their relative cellular density. PMID- 2993319 TI - Reciprocal transmembranous receptor-cytoskeleton interactions in concanavalin A activated platelets. AB - Concanavalin A (Con A) has been used to activate platelets, inducing a specific interaction between the glycoprotein IIb-IIIa complex and the cytoskeleton of the activated platelet. In agreement with this, we have shown that Con A activates human platelets, initiating phosphorylation, secretion, and cytoskeletal formation. Con A and cytochalasin B were used to demonstrate a reciprocal interaction of the glycoprotein complex with the platelet cytoskeleton. Additionally, we have shown that a similar reciprocity is provided by the multivalent fibrin-fibrinogen platelet interaction found in the thrombin-induced clot. Con A differs from other activators in precipitating an apparent cytoskeletal core despite a complete inhibition of platelet activation by prostaglandin E1. We suggest, from this result, that Con A may be cross-linking a membrane-associated cytoskeletal complex present in the unactivated platelet. PMID- 2993321 TI - Alpha-adrenoreceptors and muscarine receptors in human pial arteries and microvessels: a receptor binding study. AB - Human pial arteries and intraparenchymal microvessels were isolated for enzyme assays and radioligand binding studies of receptors. Special attention was paid to contamination with brain tissue, which was assessed by luxol staining and cerebroside assays for myelin and by scanning electron microscopy. The amount of contamination was approximately 1% for pial vessels and 14% for microvessel preparations. Significant levels of alpha 1-adrenoreceptors (binding sites for [3H]prazosin) and alpha 2-adrenoreceptors (sites labeled by [3H]azidoclonidine) were found in both types of vessels, suggesting that each receptor can modify contractility in these human vessels. Levels of muscarine receptors (sites labeled with [3H]quinuclidinyl benzilate) and choline acetyltransferase activity were considered significant only in pial vessels. PMID- 2993322 TI - Particle radiotherapy. PMID- 2993323 TI - Polyneuropathy. Inflammatory or toxic? PMID- 2993324 TI - Separation of large DNA restriction fragments by high-performance gel filtration on TSKgel DNA-PW. PMID- 2993325 TI - Separation of flavivirus membrane and capsid proteins by multistep high performance liquid chromatography optimized by immunological monitoring. AB - Complete separation of the three structural proteins, E, C and M, of an enveloped virus (tick-borne encephalitis virus) was achieved by means of a two-step high performance liquid chromatography (HPLC) technique in less than 1 h. The hydrophobically associated membrane proteins E and M were successfully separated by high-performance gel permeation chromatography (TSK-3000 SW column) in the presence of sodium dodecyl sulphate (SDS), whereas the separation of M and C as well as desalting and removal of SDS was achieved by subsequent reversed-phase chromatography on a C3 column. With regard to further characterization by peptide mapping, analysis of the amino acid composition and aminoterminal sequencing, the second step was performed with volatile buffer systems. Quality control of the separation was achieved by a combination of HPLC with a highly sensitive dot immunoassay by the use of polyclonal as well as monoclonal antibodies. This method proved extremely sensitive and revealed strong tailing effects and cross contaminations of peaks well-separated in reversed-phase chromatography, which were neither apparent in the absorbance curve at 214 nm nor in the analysis by SDS-polyacrylamide gel electrophoresis. By visualization of the peak-tailing effect, the chromatographic conditions could be modified in order to achieve an optimum separation of proteins. PMID- 2993326 TI - Purification of gelatin-specific neutral protease from human skin by conventional and high-performance liquid chromatography. AB - Human skin, maintained in serum-free organ culture, secretes a neutral metalloendopeptidase which is remarkably specific for gelatin. Because the product peptides from the action of collagenase on collagen become denatured into random coil polypeptides of 25000 and 75000 daltons at physiological temperature, it is thought that this "gelatinase" is the second, and possibly the only other enzyme in the pathway of extracellular collagen degradation. New types of high performance liquid chromatography (HPLC) columns have enabled us to improve the yields of active gelatinase from skin culture medium. Raw medium, which has been dialyzed and lyophilized, is fractionated with ammonium sulfate, and applied to Pharmacia Blue Sepharose in a batch step. The 0.4 M sodium chloride eluate is then subjected to gel filtration on Sephacryl S-200, followed by gradient elution from Amicon Green Sepharose. The fractions with gelatinolytic activity are applied to a Bio-Rad TSK-Phenyl-5PW HPLC column for mild hydrophobic chromatography with a gradient of decreasing ammonium sulfate concentration. In the final step, the enzyme is applied to a Pharmacia Mono-Q FPLC column and eluted with a gradient of sodium chloride. At this point, the enzyme appears as two bands, corresponding to enzymatic activity zymograms on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. PMID- 2993327 TI - Combined size-exclusion and reversed-phase high-performance liquid chromatography of a detergent extract of Sendai virus. AB - Virus envelope proteins obtained by Triton X-100 extraction of Sendai virions were purified to a high degree by a combination of high-performance liquid chromatography (HPLC) methods. Size-exclusion HPLC on a TSK 4000 PW column with several concentrations of acetonitrile or ethanol-1-butanol in 0.1% hydrochloric acid as eluent was used as the first chromatographic step. Peak fractions were diluted in water and further fractionated on reversed-phase columns (TMS-250 or Vydac 218 TP). Size-exclusion HPLC with 45% acetonitrile in 0.1% hydrochloric acid, combined with reversed-phase HPLC on either column, was most suitable for obtaining highly purified F2 protein. Antibodies obtained after injection of this protein were reactive with the intact virus. PMID- 2993328 TI - Isolation of the subunits of the coronavirus envelope glycoprotein E2 by hydroxyapatite high-performance liquid chromatography. AB - The coronavirus glycoprotein E2, which is responsible for virus attachment to cell receptors and virus-induced cell fusion, was purified by solubilization of virions with Triton X-114 and phase fractionation. Native E2 and tryptic subunits of the glycoprotein were separated by size-exclusion high-performance liquid chromatography (HPLC). Two distinct 90 kD E2 subunits, which had identical electrophoretic mobilities when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were separated by hydroxyapatite HPLC in the presence of sodium dodecyl sulfate. PMID- 2993329 TI - High-performance liquid chromatography of DNA restriction fragments. AB - High-performance liquid chromatography on Nucleogen-DEAE 4000-10 has been applied to several problems of the isolation of DNA restriction fragments. Large amounts of DNA fragments of high purity are necessary for biophysical studies and for molecular hybridization in basic research, as well as in medical diagnosis. The influence of various parameters, such as buffer, pH, eluting salt, gradient slope, flow-rate and the addition of urea on the resolution of fragments by high performance liquid chromatography were studied on an analytical scale, and the optimal conditions were then used for the large-scale preparation of milligram amounts. The best resolution of fragments between 25 and 1500 base pairs was obtained with a linear gradient from 500 mM to 1200 mM sodium chloride in 6 M urea -30 mM sodium phosphate (pH 6.0). Quantitative data are given for the purity and recovery of the sample, and the capacity and lifetime of the column. The following applications of high-performance liquid chromatography of restriction fragments are described: preparation of 2 mg of fragments, separation of 1 mg of DNA insert from 7 mg of its plasmid vector, and analysis of DNA-RNA hybrids. PMID- 2993330 TI - Comparison of reversed-phase columns for the separation of tryptic peptides by gradient elution. Correlation of experimental results and model prediction. AB - A total of 10 reversed-phase columns obtained from different suppliers were evaluated for their ability to separate typical peptide mixtures: tryptic digests of cytochrome c, lysozyme and collagen. Each column was tested in a standard gradient elution using phosphate buffer (pH 2.85) as the A-solvent and acetonitrile as the B-solvent. Some differences in band spacing for the various peptides were observed from column to column. More important differences were observed in the ability of these various columns to provide narrow peaks and good resolution, as measured by column peak capacity. These differences in column peak capacity were related to differences in column dimensions and particle size; it also appeared that C8 bonded phases were somewhat more efficient than were C18 phases. The recovery of cytochrome c digest from four of the columns was also assessed. Summed peak areas were proportional to sample size, as the latter was varied from 1 to 100 micrograms. The apparent recovery from each column did not vary by more than +/- 5%. From this it was concluded that these columns gave essentially quantitative recovery for this particular sample. Limited data on column life were obtained for some of these columns. PMID- 2993331 TI - Characterization of a solvent system for separation of water-insoluble poliovirus proteins by reversed-phase high-performance liquid chromatography. AB - Formic acid in high concentration is an extremely potent solvent for proteins, particularly for hydrophobic ones. 60% Formic acid, necessary for solubilization of structural polypeptides of poliovirus and other proteins, modified at the cysteines, was used together with 2-propanol or acetonitrile as organic modifier for gradient elution in reversed-phase high-performance liquid chromatography. Several reversed-phase columns were tested. In each case, polypeptides were eluted quantitatively. It was demonstrated that this solvent system, with its high proportion of formic acid, did not affect the size, hydrophobicity and charge of the separated polypeptides. By injection into rabbits of poliovirus polypeptides, obtained in high purity by chromatography in the new solvent system, monospecific antibodies were induced, the specificity of which was determined by immunoprecipitation. PMID- 2993332 TI - High-performance chromatofocusing of steroid receptor proteins in the presence and absence of steroid. Investigation of steroid-dependent alterations in surface charge heterogeneity. AB - We have previously reported the development of high-performance chromatofocusing (HPCF) systems for rapid evaluation of the surface charge heterogeneity of steroid receptor proteins, each in the presence of its specific steroid ligand. However, the surface charge properties of ligand-free receptor proteins remain largely unknown. We have now employed HPCF to rapidly evaluate the surface charge properties of cytosolic estrogen receptor proteins in both the presence and absence of the ligand ([3H]estradiol-17 beta). All operations were performed at 0 4 degrees C. Cytosols prepared from immature calf uteri were preparatively analyzed by HPCF on a SynChropak AX-500 column (25 cm X 4.6 mm I.D.) either before or after incubation with 5-10 nM [3H]estradiol-17 beta. Elution of receptor was by generation of internal pH gradients (pH 8.1 to 3.2) using Pharmacia Polybuffers 96 and 74. Postcolumn detection of previously unliganded receptor was accomplished by incubation of pH-neutralized (pH 7.4) fractions with 5 nM [3H]estradiol-17 beta in the presence and absence of unlabelled competitor. Specifically bound steroid was determined in each fraction using an hydroxylapatite adsorption assay. Significant surface charge heterogeneity was observed for both unliganded receptor and the steroid-receptor complex. The heterodisperse pattern of receptor surface charge appeared to vary in a steroid dependent manner. Preformed steroid-receptor complexes eluted primarily between pH 6.5-7 and between pH 5-6, with indications for heterogeneity within both regions. The surface charge distribution of unliganded receptor routinely revealed additional, more acidic eluting (pH 3.8-4.6) receptor forms. Sodium molybdate, a commonly used receptor-stabilizing agent, maintains receptors during HPCF as relatively acidic eluting forms (pH 3.8-5.0). The specific elution profile of molybdate-stabilized receptor also appears steroid-dependent. These data demonstrate that HPCF can be used preparatively to rapidly isolate unliganded receptor forms in a biologically active state. PMID- 2993333 TI - Detection of sendai virus protein by reversed-phase high-performance liquid chromatography combined with immuno-chromatography. AB - Reversed-phase high-performance liquid chromatography (RP-HPLC) was applied to the detection of Sendai virus F2 protein. Immunoaffinity chromatography was used prior to RP-HPLC to increase the specificity. With this combined method, the presence of F2 protein in complex solutions could be demonstrated within 150 min. Using RP-HPLC, F2 protein could be detected in amounts down to 20 ng, which could be reduced to 1 ng by application of a microbore column. PMID- 2993334 TI - Use of silver nitrate impregnated silica cartridges in the separation of kahweol and cafestol esters by preparative liquid chromatography. PMID- 2993335 TI - Detection of rotavirus-specific IgG antibodies by immunoperoxidase assay and enzyme-linked immunosorbent assay. AB - An indirect immunoperoxidase assay (IPA) has been developed for determination of IgG antibodies to rotavirus. The technique employed as antigen, SA-11 infected MA 104 cells, which were air-dried on glass slides and acetone-fixed. In parallel, rota-specific IgG antibodies were determined by enzyme-linked immunosorbent assay (ELISA). Specific IgG antibodies to rotavirus were determined in sera of healthy children and in sera of patients suffering from gastroenteritis. A good correlation (r = 0.92) and (r = 0.98) for healthy children and patients, respectively, was found between IPA and ELISA techniques. The IPA technique is rapid and simple and positive results, because of the intensive staining, are easily read by low-power light microscope. The potential application of IPA and ELISA methods in serodiagnosis of rotavirus infections is discussed. PMID- 2993336 TI - Detection of cytomegalovirus by DNA-DNA hybridization employing probes labelled with 32-phosphorus or biotin. AB - Various factors influencing the detection of human cytomegalovirus (HCMV) in infected cells by DNA-DNA hybridization have been investigated. Employing the Hind III O fragment of HCMV AD169 labelled with 32P, we found that detection sensitivity was highly influenced by the method employed for extraction of DNA from infected cells. Excision of the Hind III O fragment from the vector by restriction endonuclease digestion prior to 32P-labelling further improved the detection capability of the probe. Similarly, cytomegalovirus (CMV) DNA detection employing biotin-labelled probes and streptavidin/alkaline phosphatase in the hybridot assay was also highly dependent on the method of DNA extraction prior to hybridization. Finally, we describe an in situ assay employing a biotin-labelled probe and fluorescein-conjugated avidin to detect CMV DNA in cultured cells. PMID- 2993337 TI - Quantitative microcomplement fixation tests using chicken anti-viral antibody extracted from egg yolk. AB - Yolk immunoglobulins obtained from hens immunized with human rotavirus and two plant virus antigens were used in quantitative microcomplement fixation tests. Difficulties inherent in the use of avian complement were overcome by utilizing a mixture of the C1 component of chicken complement and guinea-pig complement devoid of its own C1. The method is suitable for detecting small quantities of viral antigen and antibody and for detecting antigenic activity present on peptide fragments of viral proteins. PMID- 2993338 TI - The use of a random priming procedure to generate cDNA libraries of infectious bronchitis virus, a large RNA virus. AB - An efficient method for the generation of gene banks from large RNA viruses is described using infectious bronchitis virus as an example. Randomly primed clones have been characterized and found to be representative of the viral genome including 5' leader sequences. PMID- 2993339 TI - Effects of a short course of estrogen on mineral metabolism in postmenopausal women. AB - Previous studies suggested that estrogen administration leads to an increase in circulating immunoreactive PTH (iPTH), thought to be secondary to a slight decrease in serum calcium resulting from inhibition of bone resorption. Using three different RIAs, we measured iPTH in serum from 10 postmenopausal women before and after 14 days of ethinyl estradiol administration. In 2 sensitive RIAs directed at the midregion of the PTH molecule, iPTH values fell or remained unchanged in each subject, with average decreases of 23% (P less than 0.001) and 28% (P less than 0.005) in the two assays. Total urinary cAMP, the tubular maximum for urinary phosphate excretion, and serum iPTH measured with the third RIA did not change after estrogen treatment. Fasting urinary calcium and hydroxyproline and serum calcium, phosphorus, albumin, alkaline phosphatase, and osteocalcin all decreased after treatment, and serum 1,25-dihydroxyvitamin D increased in each subject. In a second cohort of 5 women given ethinyl estradiol for 8 weeks, similar changes were found at 2 weeks, but there was a trend toward increasing serum iPTH, increasing total urinary cAMP excretion, and decreasing the tubular maximum for urinary phosphate excretion by 8 weeks. The increase in serum 1,25-dihydroxyvitamin D and the decrease in serum osteocalcin were again found after 2 weeks of estrogen and did not change further despite continued treatment. These results indicate multiple effects of a 2-week course of estrogen treatment on mineral metabolism in the absence of an increase in serum iPTH or several biological indices of PTH activity. PMID- 2993340 TI - On the mechanism of secondary hyperparathyroidism in moderate renal insufficiency. AB - The effect of prolonged (60 days) dietary phosphate restriction on divalent ion metabolism was studied in four patients with moderate renal insufficiency in an effort to delineate the mechanisms of secondary hyperparathyroidism in renal failure. The patients had low normal serum phosphorous and normal vitamin D metabolite levels but had evidence of disturbances in target organs for vitamin D, including impaired intestinal absorption of calcium, reduced calcemic response to PTH, low serum ionized calcium levels, and, consequently, elevated PTH levels. After dietary phosphate restriction, there was marked improvement or normalization of intestinal absorption of calcium, calcemic response to PTH, and ionized calcium and PTH levels. There was also a significant rise (44%) in serum 1,25-dihydroxyvitamin D levels. The data suggest that intracellular phosphorous retention, which may develop as renal insufficiency ensues, may interfere with the action and production of 1,25-dihydroxyvitamin D. This leads to defective intestinal calcium absorption and reduced calcemic responses to PTH. As a result, hypocalcemia develops, causing secondary hyperparathyroidism. Our data assign a critical role for a disturbance(s) in vitamin D metabolism in genesis of the hyperparathyroidism of renal failure. PMID- 2993341 TI - Enhancement of adrenal cortisol secretion after intravenous high dose dexamethasone. AB - To assess the possible existence of a short loop feedback mechanism of direct glucocorticoid suppression on the adrenal glands, we performed a series of tests employing insulin hypoglycemia or ACTH infusions to obtain adrenal stimulation by ACTH levels that remained within the physiological range. Although the rapidity of glucocorticoid suppression of the pituitary thwarted efforts to use endogenous ACTH as a stimulus, we were able to mimic the stressed state by very low dose ACTH infusions (0.05 microgram/kg BW). No inhibition of cortisol secretion in response to ACTH infusions was detected in tests done after administration of dexamethasone compared to placebo [mean integrated response, 29.37 +/- 1.91 (+/- SE) vs. 29.12 +/- 1.12 microgram . h/dl, respectively]. Furthermore, when high doses of dexamethasone were administered iv, a small paradoxical increase in cortisol secretion was found (33.82 +/- 1.44 microgram . h/dl) without a difference in ACTH levels. These data do not support the concept of significant direct glucocorticoid inhibition of adrenal secretion. Non-ACTH factors that may enhance cortisol secretion in the presence of ACTH may exist. PMID- 2993343 TI - Effect of synthetic human atrial natriuretic peptide on aldosterone secretion by dispersed aldosterone-producing adenoma cells in vitro. AB - The effect of synthetic alpha-human atrial natriuretic peptide (alpha hANP), a potent natriuretic and vasorelaxant polypeptide recently isolated from human atria, on aldosterone secretion was studied in vitro in collagenase-dispersed adrenal adenoma cells from a patient with primary aldosteronism. alpha hANP (3.2 X 10(-7) M) significantly inhibited both basal and potassium (16 mM)-stimulated aldosterone secretion, whereas it had little or no effect on aldosterone secretion submaximally or maximally stimulated by ACTH (3.4 X 10(-10)-3.4 X 10( 9) M) or angiotensin II (10(-8)-10(-9) M). The less potent effect of alpha hANP on aldosterone secretion by dispersed human adrenal tumor cells compared to that in in vitro animal studies may reflect decreased affinity and/or number of specific receptors for ANP on the tumor cells. Whether ANP plays a physiological role in regulation of aldosterone secretion in humans in vivo remains to be determined. PMID- 2993342 TI - Evidence from monoclonal antibody studies that insulin stimulates deoxyribonucleic acid synthesis through the type I somatomedin receptor. AB - Somatomedin-C/insulin-like growth factor-I (Sm-C/IGF-I) and insulin stimulate DNA synthesis and cell replication in cultured human fibroblasts. It has been postulated that the growth-promoting actions of both peptides are mediated through the type I Sm-C/IGF-I receptor. This study tests this hypothesis using two recently developed monoclonal antibodies. The antibody designated sm 1.2 is directed to Sm-C, whereas the antibody designated alpha IR-3 is directed against the type I receptor for Sm-C/IGF-I. Radiolabeled monoclonal antibody alpha IR-3 was bound to human foreskin fibroblasts in a reversible time-dependent fashion, with 90% of the specific binding complete after 6 h of incubation at 15 C. Binding of [125I]alpha IR-3 was completely inhibited by excess unlabeled antibody, but not by 50 nM Sm-C or 1000 nM insulin. Specific binding of [125I]Sm C fell to 27% of the control value in the presence of 50 nM alpha IR-3, and this concentration of antibody significantly reduced the mitogenic response to both Sm C and insulin. Antibody sm 1.2 blocked the mitogenic response to exogenous Sm-C, but did not block the response to insulin; indeed, in some experiments, sm 1.2 enhanced the response to insulin. We postulate that this enhancement is the result of neutralizing endogenously produced Sm-like substances. This study provides further evidence that the growth-promoting effects of insulin in this cell type are the result of interaction with the Sm-C/IGF-I receptor. PMID- 2993344 TI - Adrenocorticotropin-stimulated adrenal androgen secretion in anorexia nervosa: impaired secretion at low weight with normalization after long-term weight recovery. AB - Adrenal androgen secretion is decreased in patients with anorexia nervosa. To assess the reversibility of the decreased secretion with recovery of body weight, we measured ACTH-stimulated adrenal androgen levels at different stages of recovery. Basal plasma GH and somatomedin-C levels also were measured, because both have been proposed as potential stimuli for adrenal androgen secretion. When studied at low body weight [58 +/- 3% (+/- SEM) ideal BW], women with anorexia nervosa had decreased ACTH-stimulated levels of dehydroepiandrosterone (DHA), DHA sulfate (DHAS), and androstenedione and decreased DHA to cortisol, DHAS to cortisol, and androstenedione to cortisol ratios compared to normal women. Women who had recently completed a refeeding program (within 2-4 weeks, 81 +/- 2% ideal BW) had an increased somatomedin-C level compared to low weight patients, but similar ACTH-stimulated adrenal androgen levels. Long term weight-recovered women (86 +/- 4% ideal BW, recovery for more than 6 months, with resumption of menses), however, had significant increases in ACTH-stimulated DHA and DHAS levels and DHA to cortisol and DHAS to cortisol ratios, and their hormone levels and ratios were not different from those in normal women. GH levels fell during weight recovery, although the values in the three patient groups did not differ significantly. We conclude that the recovery of adrenal androgen secretion while GH levels were falling provides evidence against a direct effect of GH as a stimulus for adrenal androgen secretion. The recovery of somatomedin-C before the recovery of adrenal androgens, however, and the positive correlation between plasma somatomedin-C and the integrated level of plasma DHAS (r = 0.50; P less than 0.02) are consistent with the hypothesis that somatomedin-C is a stimulus for adrenal androgen secretion. PMID- 2993345 TI - Treatment of prostatic cancer with a gonadotropin-releasing hormone agonist analog: acute and long term effects on endocrine functions of testis tissue. AB - Six patients with advanced prostatic cancer were treated with a potent GnRH agonist analog (buserelin, Hoechst; 600 micrograms, intranasally, three times a day) for 6 months. The first 2-6 days of treatment were associated with a 50% elevation (P less than 0.01) in serum testosterone (T) and a simultaneous elevation of 20% (P less than 0.01) in serum prostate-specific acid phosphatase (PAP). Serum T fell to the castrate range (less than 1 nmol/liter) in all patients in 2-3 weeks, and PAP decreased concomitantly in five of the six patients. Serum LH progressively decreased by about 80% during the treatment, whereas a secondary rise of FSH levels occurred after the first month of treatment. The patients were orchiectomized after 6 months of treatment. No differences were found between the pre- and postsurgical levels of serum T or in comparison with those of six patients orchiectomized as the first therapeutic measure. Testicular endogenous T concentrations, LH and FSH receptors, and in vitro T production were measured in three testis samples and compared with those values in testis tissue obtained from five control patients. The endogenous levels and in vitro production of T were depressed by over 95% in testes from agonist-treated patients. The small residual T production responded to hCG stimulation as in control patients. Interestingly, no change was found in testicular LH receptor content, but FSH receptors decreased by 80%. The elevation in serum PAP at the beginning of the agonist treatment and the small residual testicular T production after 6 months may not be clinically important. However, they indicate the necessity of comparative long term studies between orchiectomy and GnRH agonists in the treatment of patients with prostatic cancer. PMID- 2993346 TI - Distribution and characterization of insulin and insulin-like growth factor I receptors in normal human ovary. AB - Insulin-resistant hyperinsulinemic states are now widely known to be associated with ovarian hyperandrogenism, and this is thought to be due to an action of insulin on the ovary. However, the identity of the receptor that is responsible for insulin action in these patients, whose insulin receptors on classical target tissues are severely impaired, is unclear. We now report the presence of insulin receptors in stromal and follicular compartments as well as in granulosa cells obtained from normal ovaries. After 15-h incubations at 4 C with [125I]insulin and tissue fragments, specific insulin binding was 6-19% and 7-13%/mg protein (n = 8) to stroma and theca, respectively. Granulosa cells obtained in the course of in vitro fertilization were separated from red cells on a Percoll gradient; specific insulin binding ranged from 9-15%/10(6) cells. Insulin binding was characterized by sensitive insulin competition (half-maximal, 10 ng/ml), appropriately shifted proinsulin competition (20 times to the right), and complete inhibition by specific anti-insulin receptor antibodies (B-2). An antibody to the insulin-like growth factor I (IGF-I) receptor (alpha IR-3) that inhibits IGF-I binding to IGF-I receptors in other cell systems had no effect on insulin binding. Further proof that this binding is to classic insulin receptors was obtained from measurement of insulin-stimulated receptor autophosphorylation. When insulin receptors from stroma were extracted with Triton X-100 and incubated with [gamma-32P]ATP and Mn, insulin increased the incorporation of 32P into the beta-subunit of the receptor 5-fold. In parallel studies with [125I-]IGF-I and specific blocking antibodies to its receptor, no detectable IGF-I binding to stroma or follicles was found. We conclude that specific high affinity insulin receptors possessing tyrosine kinase activity are widely distributed in normal human ovary. IGF-I receptors in normal ovary are either absent or present at very low density. Binding of insulin to its own receptor (as opposed to IGF-I receptors) appears to be the most likely first step in the stimulation of ovarian steroidogenesis by insulin in normal ovaries and possibly in insulin-resistant states as well. PMID- 2993348 TI - Thyrotropin receptor-adenylate cyclase system in Hurthle cell neoplasms. AB - Hurthle cell neoplasms are though to arise from the follicular cells of the thyroid gland, although some studies suggest that they originate from the parafollicular cells. We studied tissue from five patients (three men and two women, aged 33-72 yr) with Hurthle cell neoplasms (four adenomas and one carcinoma) to determine whether Hurthle cell neoplasms have an intact TSH receptor-adenylate cyclase (AC) system and, if so, whether it differs in benign and malignant Hurthle cell and neoplasms. Binding of [125I]bovine (b) TSH and an AC response to TSH occurred in all four Hurthle cell adenomas. In three of these tumors, there was good binding and a relatively good correlation between the concentration of bTSH producing half-maximal inhibition of [125I]bTSH binding (4.8 mU/ml) and the TSH concentration causing half-maximal stimulation of AC (2.2 mU/ml). There was also a 2- to 5-fold increase in AC activity in response to bTSH (300 mU/ml), a value comparable to that which occurs in follicular thyroid neoplasms and differentiated thyroid cancers. In the fourth Hurthle cell adenoma, however, binding was low, the apparent Kd (170 mU/ml) and Km (20 mU/ml) were high, and there was only a 1.4-fold increase in AC activity in response to bTSH (300 mU/ml). In the one metastatic Hurthle cell carcinoma, there was no high affinity TSH binding or AC response to TSH. Thus, Hurthle cell neoplasms are of follicular cell origin, since benign Hurthle cell tumors have an intact TSH receptor-AC system. Malignant Hurthle cell neoplasms, like undifferentiated and medullary thyroid cancer, lack a functional TSH receptor. PMID- 2993347 TI - Identification of 1,24(R)-dihydroxyvitamin D3-like bone-resorbing lipid in a patient with cancer-associated hypercalcemia. AB - A lipid indistinguishable from 1,24(R)-dihydroxyvitamin D3 [1,24(R)-(OH)2D3] was found in serum and tumor extracts from a hypercalcemic patient with a small cell carcinoma of the lung. The lipid comigrated with authentic 1,24(R)-(OH)2D3 on high performance liquid chromatography using both straight and reverse phase columns and competed with tritiated 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3)] for binding to intestinal 1,25-(OH)2D3 receptor. Increasing doses of the lipid factor from tumor and authentic 1,24(R)-(OH)2D3 gave parallel responses in a bone resorption assay, as assessed by 45Ca release from prelabeled mouse calvaria. The lipid factor from the patient's serum and authentic 1,24(R)-(OH)2D3 had identical biological activities in the receptor binding and bone resorption assays. In addition, the mechanisms of action of this lipid factor and 1,24(R)-(OH)2D3 were indistinguishable. Bone resorption by both was inhibited by calcitonin, and neither the lipid factor nor authentic 1,24(R)-(OH)2D3 affected cAMP content in osteoblast-like bone cells derived from mouse calvaria. The estimated concentrations of the 1,24(R)-(OH)2D3-like lipid, expressed as 1,24(R)-(OH)2D3 were 11 ng/g tumor wet wt by the receptor binding assay and 9.2 ng/g tumor wet wt by the bone resorption assay. The mean serum concentration was 1.4 +/- 0.3 (+/- SD) ng/ml (n = 3) by the receptor binding assay. No activity was detected in either bioassay when extracts of nontumor tissues from this patient or tumor extracts and sera from one hypercalcemic and four normocalcemic cancer patients were tested. The mean serum 1,25-(OH)2D level was low (6.4 +/- 0.5 pg/ml; n = 2), and serum 1,24(R),25-(OH)3D in this patient was high (103 pg/ml) compared to normocalcemic cancer patients, in whom the mean serum 1,25-(OH)2D level was 27 +/ 12 pg/ml (n = 4) and the 1,24(R),25(OH)3D level was 28 +/- 1.3 pg/ml (n = 4). Thus, the 1,24(R)-(OH)2D3-like lipid may be a substrate for metabolic conversion to 1,24(R),25-(OH)3D in vivo. These results provide evidence for the presence of a novel metabolite of vitamin D3, 1,24(R)-(OH)2D3. Detection of this bone resorbing lipid in both tumor and serum suggests, but does not prove, that the tumor secreted this bioactive lipid into the circulation and that the high level of circulating bone-resorbing lipid was related to the hypercalcemia in this patient. PMID- 2993350 TI - Selection of nonfastidious adenovirus species in 293 cells inoculated with stool specimens containing adenovirus 40. AB - Of 35 stool specimens isolated and examined in 293 cells, 15 isolates contained adenovirus species 40 (Ad40), and 4 of these 15 isolates also contained a nonfastidious adenovirus species (Ad1 in two cases, Ad18 or Ad31) which was selected over Ad40 during serial passage in the 293 cells. The selection of Ad1 over Ad40 was examined in detail. Restriction analysis of intracellular DNA and the relative infectivity titers of Ad40 and Ad1 at each passage level after the inoculation of 293 cells with a particular stool specimen demonstrated that although the amount of Ad40 DNA synthesized far exceeded that of Ad1, the relative infectivity titer of Ad40 was low. The growth characteristics of Ad40 were then compared with those of Ad1, Ad18, and Ad41 in singly infected 293 cell cultures. One-step growth curves showed the same growth rate in each case, with a latent period of 12 h and a maximum titer at 24 to 36 h postinfection. Yields of infectious Ad40 virus were consistently 100- to 1,000-fold lower than those of Ad1. This difference was reflected by a reduced yield of total AD40 virions (p1.34) as determined by 35S labeling experiments. However, the 3- to 10-fold reduction in total yield of Ad40 virions did not account for the 100- to 1,000 fold reduction in the yield of infectious virus. PMID- 2993349 TI - Isolation and differentiation of herpes simplex virus and Trichomonas vaginalis in cell culture. AB - During the period January 1982 to January 1985, 2,234 specimens were cultured for isolation of herpes simplex virus (HSV). HSV was isolated from 23% of these, Trichomonas vaginalis was isolated from 1.6%, and 75.3% were negative. In 0.2% of these, HSV and T. vaginalis were isolated from the same specimen. Cytopathic effects produced by HSV were identified by their sensitivity to arabinosylthymine, whereas those produced by T. vaginalis were identified by their lack of sensitivity to arabinosylthymine and by observation of motility. Cytopathic effects produced by T. vaginalis were reproduced by trophozoites from axenic cultures of T. vaginalis as well as by lysates of T. vaginalis added to serum-free BHK cells. PMID- 2993351 TI - Factors affecting the detection of enteroviruses in cerebrospinal fluid with coxsackievirus B3 and poliovirus 1 cDNA probes. AB - Enteroviruses are common pathogens of meningitis and encephalitis, and infections are often difficult to distinguish clinically from bacterial and herpetic infections of the central nervous system. An array of enteroviruses added to cerebrospinal fluid in reconstruction experiments were detected by a dot hybridization assay. Optimal handling and processing conditions for infected cerebrospinal fluid were established, and the effect on the hybridization reaction of humoral and cellular components of the inflammatory response was determined. Six hybridization probes, derived from poliovirus 1 and coxsackievirus B3, were then tested, singly and in combinations, to optimize the sensitivity and spectrum of the assay. Implications for enteroviral taxonomy based on these experiments are discussed. PMID- 2993352 TI - Retrospective evaluation of the isolation and identification of herpes simplex virus with Cultureset and human fibroblasts. AB - A total of 442 specimens from various anatomic sites was cultured for herpes simplex virus during the past 2 years. Most specimens were obtained from the respiratory tracts and cutaneous lesions of immunocompromised hosts (232 specimens) or the female genital tract (138 specimens). Two tubes containing human newborn foreskin fibroblasts and two Ortho Cultureset tubes containing Vero cells were inoculated with each specimen. The 384 inoculated specimens were stained with Cultureset peroxidase-antiperoxidase reagents within 48 h and again at 3, 4, or 5 days if initially negative. Fibroblasts were inspected for cytopathic effect for 7 days. Of these 384 specimens, Cultureset detected 57 of 62 positive specimens within 48 h; fibroblasts detected 58 positive specimens by 7 days. The calculated sensitivity and specificity for Cultureset at 48 h were 91.9 and 100%, respectively. However, when all results were considered, including those that became positive in Cultureset after 48 h, the calculated sensitivity and specificity for Cultureset were 98.8 and 100%, respectively. We conclude that Cultureset is a reliable method for detection of herpes simplex virus when two tubes are inoculated and stained as described. PMID- 2993353 TI - Detection of Norwalk virus antibodies and antigen with a biotin-avidin immunoassay. AB - Biotin-avidin immunoassays (BAIs) were developed to detect Norwalk virus antigen and to measure Norwalk virus antibody. The BAI detected Norwalk virus infections by a fourfold titer rise in antibody in sera or by antigen in stool, with a sensitivity similar to or greater than that of the radioimmunoassay (RIA), and the BAI appeared to be more sensitive than the RIA for detecting antibody in single serum specimens. The BAI antigen test detected Norwalk antigen in all stools that were positive by the RIA, and in two stools negative by the RIA. Of 106 serum pairs tested by both the RIA and BAI, 94 demonstrated seroconversion, or lack of seroconversion, in both assays; 12 gave discordant results. Titers by the BAI generally exceeded those by the RIA by two- or fourfold. The BAI had advantages over the RIA in that it had added sensitivity in the detection of Norwalk antibody, was less time consuming, did not require frequent labeling of antibody, and did not have the problems associated with the use of isotopes. PMID- 2993355 TI - Sodium polyanetholesulfonate in the identification of Gardnerella vaginalis. PMID- 2993354 TI - Specific enzyme-linked immunoassay for rotavirus serotypes 1 and 3. AB - We prepared monoclonal antibodies against two serotypically distinct rotavirus strains: Wa, a serotype 1 virus of human origin, and rhesus rotavirus, a simian serotype 3 virus. Monoclonal antibodies which react specifically with VP7 of each serotype were identified by hemagglutination inhibition tests, plaque reduction neutralization studies, and solid-phase immunoassays which used wild-type and reassortant strains of rotavirus. An enzyme-linked immunoassay was designed which utilizes two of these antibodies to correctly identify serotype 1 and serotype 3 viruses. PMID- 2993356 TI - Crossbridge behaviour during muscle contraction. AB - A number of recent observations by probe and X-ray methods on the behaviour of crossbridges during contraction is considered in relation to the energetics of the process. It is shown that a self-consistent picture of the crossbridge cycle, compatible with these observations and involving strongly and weakly attached crossbridges, can be obtained providing that the tension-generating part of the crossbridge stroke is only about 40 A i.e. about one-third of the usually accepted value. The myosin head subunits in the tension-generating bridges could have a configuration close to that of rigor. A mechanism is suggested whereby rapid tension recovery after quick releases up to 120 A could still be produced by such a system. PMID- 2993357 TI - Activities for injury prevention. PMID- 2993358 TI - Tumor products and the hypercalcemia of malignancy. PMID- 2993359 TI - Interleukin-2 receptor (Tac antigen) expressed on adult T cell leukemia cells. AB - We studied the expression of the interleukin-2 (IL-2) receptor and the proliferative response to exogenous IL-2 of peripheral blood leukemic cells from patients with adult T cell leukemia (ATL) in order to see whether IL-2 receptor expressed on ATL cells is different from normal IL-2 receptor and whether it plays a role in the neoplastic growth in ATL. Peripheral blood leukemic cells from 42 patients with ATL examined expressed IL-2 receptors that were detected by anti-Tac monoclonal antibody when examined immediately after the separation of cells or after the culture for 24 or 48 h. The number of anti-Tac binding sites ranged from 3,100 to 11,400 in fresh cells and from 3,600 to 96,000/cell in short term cultured leukemic cells, whereas phytohemagglutinin-P (PHA-P)-stimulated normal T cells exhibited 6,900-35,000 anti-Tac binding sites per cell. ATL derived and human T cell leukemia/lymphoma virus, type I (HTLV-I)-infected cell lines such as MT-1 and Hut102 expressed a much higher number of anti-Tac binding sites. Leukemic cells from 15 patients with ATL examined showed no or very poor proliferative response to various concentrations of immunoaffinity-purified IL-2, although they expressed Tac antigen (Ag). Radiolabeled IL-2 binding experiments demonstrated that ATL leukemic cells could bind IL-2, and they expressed both high and low affinity IL-2 receptors, although the number of high affinity IL-2 receptor was much less than that of low affinity IL-2 receptor and that of anti Tac binding sites. In contrast, leukemic T cells from a patient with T cell chronic lymphocytic leukemia (CLL), in whom HTLV-I infection was not demonstrated, responded as well as PHA-P-stimulated normal T cells, and their IL 2 receptors, unlike ATL cells, were modulated (down regulated) by anti-Tac antibody. No differences were noted between ATL cells and normal activated T cells in one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the IL-2 receptor. Thus, leukemic cells in ATL spontaneously and continuously express IL-2 receptor, which appears to be abnormally regulated and unresponsive to IL-2. These results, taken together with those on normal IL-2 receptors on HTLV-I-negative T-CLL cells, suggest that abnormal expression of the IL-2 receptor in ATL is closely associated with HTLV-I infection and may play a role in the neoplastic growth of ATL cells. PMID- 2993360 TI - Evidence for beta adrenoceptors in proximal tubules. Isoproterenol-sensitive adenylate cyclase in pars recta of canine nephron. AB - Observations in vivo suggest that catecholamines modulate reabsorptive functions of proximal tubules by acting on beta-adrenoceptors. However, beta-catecholamine binding sites or beta-adrenoceptor-sensitive adenylate cyclase (AdC) has not been found in segments of proximal tubules of rat, rabbit, or mouse kidney. In the present study, we investigated the responsiveness of AdC to catecholamines, [8 Arg]vasopressin (AVP), and to parathyroid hormone (PTH) in proximal convoluted tubules (PCT), proximal straight tubules (PST), and in late distal convoluted tubules (LDCT) microdissected from canine kidney. Isoproterenol (ISO) caused a marked and dose-dependent stimulation of AdC in PST (maximum: delta + 850%; half maximum stimulation at 10(-7) M ISO), but ISO had no effect on AdC in PCT. The AdC in both PCT and PST was markedly stimulated by PTH; AVP stimulated the AdC in LDCT but not in PST or in PCT. The stimulatory effect of 10(-5) M ISO in PST (delta + 725%) was significantly greater than in LDCT (delta + 307%); norepinephrine and epinephrine had stimulatory effects in PST similar to ISO. The stimulation of AdC in PST by ISO was blocked by propranolol and by beta 2-blocker ICI-118551. On the other hand, alpha-blocker phentolamine and beta 1-blocker metoprolol did not abolish the stimulation of AdC in PST by ISO. The accumulation of cAMP in intact PCT and PST incubated in vitro was stimulated by PTH both in PST and in PCT, but ISO elevated cAMP (delta + 683%) only in PST. Our results show that proximal tubules of canine nephron, PST but not PCT, contain beta adrenoceptors of beta 2 subtype coupled to AdC. These observations provide direct evidence that the effects of catecholamines, either released from renal nerve endings or arriving from blood supply, can act directly on beta 2-adrenoceptors located in proximal tubules, and also suggest that at least some of the catecholamine effects in proximal tubules are mediated via cAMP generation. PMID- 2993361 TI - In vivo and in vitro studies of urinary concentrating ability in potassium depleted rabbits. AB - The factors responsible for the urinary concentrating defect associated with the potassium-depleted (KD) state are uncertain. The present studies were designed to, first, determine whether a urinary concentrating defect exists in potassium depleted rabbits and, second, to use the technique of in vitro perfusion to evaluate directly the antidiuretic hormone (ADH) responsiveness of cortical collecting tubules (CCT) in this setting. Feeding female New Zealand White rabbits a potassium-deficient diet for 2 wk caused a significant fall in plasma potassium levels in both the ad-libitum and controlled water intake groups (P less than 0.001). Muscle potassium content after 2 wk of potassium restriction fell from 45.6 +/- 0.9 to 29.0 +/- 1.2 meq/100 g fat-free dry solids (P less than 0.001). Renal papillary sodium content fell significantly from a control value of 234.6 +/- 8.0 to 182.46 +/- 10.0 meq/kg H2O after 2 wk of potassium restriction. Maximal urinary osmolality measured after 12 h of dehydration and 1.25 U pitressin IM was significantly decreased in rabbits after 2 wk of potassium restriction in both the ad-libitum and controlled water intake groups (P less than 0.001). The relationship between plasma potassium concentration and maximum urinary osmolality was significantly correlated in both the ad-libitum and controlled water intake groups, r = 0.73 and 0.68 (P less than 0.001), respectively. In addition, refeeding KD rabbits with normal chow for 1 wk resulted in normalization of both plasma potassium levels and urinary concentrating ability. CCT from control and KD rabbits were perfused in vitro at 25 degrees C. The hydraulic conductivity coefficient, Lp, was significantly reduced at all doses of ADH tested in tubules from KD rabbits when compared with control tubules. In addition, the maximal hydraulic conductivity in tubules from KD rabbits when tested with 200 microU/ml ADH at 37.5 degrees C was only 23% of control values (P less than 0.05). Furthermore, this reduced ADH responsiveness persisted when the bath potassium was elevated from 5 to 20 mM. The reflection coefficient for NaCl when compared with raffinose was 0.91 in tubules from KD animals. Thus, these data suggest that the ADH-resistant urinary concentrating defect associated with potassium depletion is due, at least in part, to a diminished responsiveness of the CCT to ADH. Therefore, further studies were designed to investigate the cellular steps involved in this abnormal response. There was no difference in the 8-para-chlorophenylthio cyclic AMP induced hydroosmotic response between CCT from KD and control rabbits. Since the cAMP induced hydroosmotic response was similar between KD and control CCT, experiments were performed to evaluate the contribution of phosphodiesterase (PDIE) activity by using the potent PDIE inhibitor isobutylmethylxanthine (10(-4) and 10(-3)M) in the presence of ADH (200 U/ml). Although Lp was increased by PDIE inhibition in CCT from both control and KD animals, the overall hydroosmotic response in CCT from KD rabbits was still significantly reduced when compared with controls. The final experiments used forskolin to evaluate further the adenylate cyclase complex. The resulting hydroosmotic response in CCT from KD rabbits was almost identical to that obtained in controls. In conclusion, these data suggest that the decreased responsiveness of CCT from KD rabbits to ADH involves a step at or proximal to the stimulation of the catalytic subunit of adenylate cyclase, and that PDIE activity makes no contribution to this abnormal hydroosmotic response. PMID- 2993362 TI - Control of glomerular hypertension limits glomerular injury in rats with reduced renal mass. AB - Micropuncture and morphologic studies were performed in four groups of male Munich-Wistar rats after removal of the right kidney and segmental infarction of two-thirds of the left kidney. Groups 1 and 3 received no specific therapy. Groups 2 and 4 were treated with the angiotensin I converting enzyme inhibitor, enalapril, 50 mg/liter of which was put in their drinking water. All rats were fed standard chow. Groups 1 and 2 underwent micropuncture study 4 wk after renal ablation. Untreated group 1 rats exhibited systemic hypertension and elevation of the single nephron glomerular filtration rate (SNGFR) due to high average values for the mean glomerular transcapillary hydraulic pressure difference and glomerular plasma flow rate. In group 2 rats, treatment with enalapril prevented systemic hypertension and maintained the mean glomerular transcapillary hydraulic pressure gradient at near-normal levels without significantly compromising SNGFR and the glomerular capillary plasma flow rate, as compared with untreated group 1 rats. Groups 3 and 4 were studied 8 wk after renal ablation. Untreated group 3 rats demonstrated persistent systemic hypertension, progressive proteinuria, and glomerular structural lesions, including mesangial expansion and segmental sclerosis. In group 4 rats, treatment with enalapril maintained systemic blood pressure at normal levels over the 8-wk period and significantly limited the development of proteinuria and glomerular lesions. These studies suggest that control of glomerular hypertension effectively limits glomerular injury in rats with renal ablation, and further support the view that glomerular hemodynamic changes mediate progressive renal injury when nephron number is reduced. PMID- 2993363 TI - Pathophysiology of chronic tubulo-interstitial disease in rats. Interactions of dietary acid load, ammonia, and complement component C3. AB - The human end-stage kidney and its experimental analogue, the remnant kidney in the rat, exhibit widespread tubulo-interstitial disease. We investigated whether the pathogenesis of such tubulo-interstitial injury is dependent upon adaptive changes in tubular function and, in particular, in ammonia production when renal mass is reduced. Dietary acid load was reduced in 1 3/4-nephrectomized rats by dietary supplementation with sodium bicarbonate (NaHCO3), while control rats, paired for serum creatinine after 1 3/4 nephrectomy, were supplemented with equimolar sodium chloride. After 4-6 wk, NaHCO3-supplemented rats demonstrated less impairment of tubular function as measured by urinary excretory rates for total protein and low molecular weight protein and higher transport maximum for para-aminohippurate per unit glomerular filtration rate, less histologic evidence of tubulo-interstitial damage, less deposition of complement components C3 and C5b-9, and a lower renal vein total ammonia concentration. Such differences in tubular function could not be accounted for simply on the basis of systemic alkalinization, and differences in tubular injury could not be ascribed to differences in glomerular function. Because nitrogen nucleophiles such as ammonia react with C3 to form a convertase for the alternative complement pathway, and because increased tissue levels of ammonia are associated with increased tubulo interstitial injury, we propose that augmented intrarenal levels of ammonia are injurious because of activation of the alternative complement pathway. Chemotactic and cytolytic complement components are thereby generated, leading to tubulo-interstitial inflammation. Thus, alkali supplementation reduces chronic tubulo-interstitial disease in the remnant kidney of the rat, and we propose that this results, at least in part, from reduction in cortical ammonia and its interaction with the alternative complement pathway. PMID- 2993364 TI - Localization of collagenase in the growth plate of rachitic rats. AB - In the transition from proliferation to hypertrophic cell zones in the growth plate, there is an increase in chondrocyte volume and a corresponding decrease in collagen content to accommodate the enlarging cells. It is postulated that collagenase accounts for this collagen loss. To test this hypothesis, tibial growth plates were obtained from normal rats, rachitic rats deficient in vitamin D and phosphate, and rats after 48 and 72 h of healing from rickets. Collagenase was quantitated by a pellet assay based on the release of solubilized collagen from the endogenous insoluble collagen in the tissue homogenates. A fourfold greater collagen release and a concomitant sixfold greater hypertrophic cell volume were measured in rachitic growth plates compared with normal age-matched controls. During healing of rickets, collagenase activity and hypertrophic cell volume returned almost to control levels. Rachitic growth plates were dissected into the juxtaepiphyseal 1/3 and the juxtametaphyseal 2/3. The latter portion contained greater than 95% of the hypertrophic cells and 86% of the collagenase. The collagen-degrading activity was extracted from this region and was shown to be a true collagenase by its production of typical A fragments of tropocollagen produced by collagenase action. The enzyme was activated by aminophenylmercuric acetate and trypsin and was inhibited by EDTA, 1,10-phenanthroline, and a tissue inhibitor of metalloproteinases from human articular cartilage. Inhibitors of aspartic, cysteine, and serine proteases had no effect. Micropuncture fluids aspirated from rachitic cartilage contained latent collagenase activity, indicating an extracellular localization. Negative tests for hemoglobin in the rachitic cartilage samples indicated that there was no contamination by capillaries and that this was not a source of collagenase. It is concluded that extracellular collagenase accounts for the loss of cartilage matrix in the hypertrophic zone, and that this process may be distinct from that of capillary invasion. PMID- 2993366 TI - Thermal inactivation of the acquired immunodeficiency syndrome virus, human T lymphotropic virus-III/lymphadenopathy-associated virus, with special reference to antihemophilic factor. AB - The virus that causes the acquired immunodeficiency syndrome (AIDS), human T lymphotropic virus/lymphadenopathy-associated virus (HTLV-III/LAV), was incubated at temperatures from 37 degrees to 60 degrees C and virus titer (ID-50) was determined over time by a microculture infectivity assay. The rate of thermal decay was consistent with first-order kinetics, and these data were used to construct a linear Arrhenius plot (r = 0.99), which was used to determine inactivation time as a function of temperature. In the liquid state, thermal decay was little affected by matrix (culture media, serum, or liquid Factor VIII). In the lyophilized state, the time required to reduce virus titer 10-fold (1 log) at 60 degrees C was 32 min compared with 24 s in the liquid state. HTLV III/LAV in liquid antihemophilic Factor VIII or IX was lyophilized and heated according to commercial manufacturers' specifications. Infectious virus was undetectable with these regimens. Heat treatment should reduce or stop transmission of HTLV-III/LAV by commercial antihemophilic Factor VIII or IX. PMID- 2993367 TI - L6 cells as a tissue culture model for thyroid hormone effects on skeletal muscle metabolism. AB - L6 cells have been investigated as a potential tissue culture model for the study of thyroid hormone effects on skeletal muscle metabolism. Differentiated L6 myotubes contained high-affinity triiodothyronine (T3) receptors with a Kd of 3 X 10(-10) M and a maximal binding capacity of 24 fmol T3/100 micrograms DNA. Undifferentiated cells contained receptors with the same Kd, but the binding capacity was reduced by at least a factor of three. Sarcoplasmic reticulum vesicle calcium pumping was demonstrated in L6 cell homogenates. The Vmax for calcium pumping was increased 2.5-fold when T3 was present in the culture medium, but the Kd was unchanged. L6 cells contained high affinity thyroid hormone receptors and were thyroid hormone responsive. These cells may be useful as a tissue culture model for studying the effects of thyroid hormones on skeletal muscle metabolism. In addition, the increase in T3 receptor number with the differentiated state suggests this as a model system for studying regulation of T3 receptor number and the role of T3 in the induction or maintenance of the differentiated state. PMID- 2993365 TI - Influences of gamma interferon on synovial fibroblast-like cells. Ia induction and inhibition of collagen synthesis. AB - The shape and function of adherent cells cultured from rheumatoid synovial membranes are influenced by immune cells, and their products. The synovial cells produce collagenase and prostaglandin E2 (PGE2), the levels of which are increased when the cells are incubated with the monokine, mononuclear cell factor/interleukin 1. The majority of adherent synovial cells are fibroblastlike in appearance and synthesize collagens and fibronectin; the synthesis of collagens and fibronectins are also increased by a monocyte factor. In the present study we found that the fibroblastlike cells expressed major histocompatibility complex class II (Ia-like) antigens after initial dispersion from the synovial membrane. Monocyte lineage antigens were detected on some round cells in early passage, but no T lymphocytes were identified in established cultures. There was loss of Ia expression on the fibroblastlike cells with age and passage in culture. The addition of the lymphokine, gamma interferon (recombinant), induced class II antigen (DR and DS/DQ) expression in early or late passage cells in a time- and dose-dependent manner and required protein synthesis. Furthermore, the adherent synovial fibroblastlike cells continued to be Ia-positive when examined as long as 10 d after the removal of gamma interferon. Ia expression was also induced by gamma interferon in normal skin fibroblasts. Synovial cells that could be induced to express Ia also bound a monoclonal antibody to type III collagen (a fibroblast marker). Gamma interferon, while inducing Ia expression, decreased the binding of type III collagen antibody on unstimulated as well as monokine-stimulated cells. Analysis of [3H]proline labeled medium by SDS polyacrylamide gel electrophoresis showed that gamma interferon decreased the synthesis of type I and III collagens and fibronectin by adherent synovial cells in a dose-dependent manner. These findings suggest that Ia expression by synovial tissue cells is not cell-specific, but reflects one or several related events, such as the degree of T lymphocyte infiltration, the presence of factors that stimulate gamma interferon release, or an increased sensitivity of the cells to gamma interferon. Whereas the synthesis of class II antigens is enhanced by the lymphokine gamma interferon, and a monocyte factor(s) stimulates collagen, collagenase and PGE2 synthesis by the same cells, gamma interferon inhibits basal and monokine-induced collagen synthesis. Thus, lymphokines and monokines may influence the extent of fibrosis as contrasted to matrix destruction at various stages of the rheumatoid lesion by affecting the function of fibroblastlike synovial cells. PMID- 2993369 TI - The calcified target lesion: mucinous carcinoma of the stomach. PMID- 2993368 TI - Muscarinic receptors on intact human fibroblasts. Absence of receptor activity in adult skin cells. AB - The intact human fibroblast has been used in clinical and basic research studies of receptor-mediated control of cell function, however there is little information about the relationship between muscarinic receptor density and the regulation of cyclic AMP (cAMP) accumulation. We have compared the muscarinic receptor characteristics of both lung and skin intact fibroblasts at fetal and adult stages of development using carbachol-mediated inhibition of cAMP accumulation and the binding of [3H]quinuclidinyl benzilate (QNB). Prostaglandin E1 (PGE1) stimulated cAMP accumulation in all four cell lines, while carbachol inhibited cAMP accumulation only in the fetal lung, adult lung, and to a lesser extent, the fetal skin. Adult skin fibroblasts did not display significant evidence of inhibitory muscarinic receptor activity. [3H]QNB binding was saturable for the fetal and adult lung, and the fetal skin, yielding Kd values of approximately 0.5 nM for these cell lines. Bmax values were 360 fmol/mg for fetal skin, 600 fmol/mg for adult lung, and 876 fmol/mg for the fetal lung. Specific binding of [3H]QNB to adult skin fibroblasts was found to be low and variable. Comparisons of the Bmax values and maximal inhibitory capacities showed that the receptor density paralleled receptor activity in all cell lines. The lack of muscarinic receptor activity in the adult skin fibroblast was confirmed in several different adult skin cell lines, indicating that the adult skin fibroblast may not be an appropriate model for muscarinic receptor activity in clinical investigations. PMID- 2993370 TI - Comparison of prevalence of human papillomavirus antigen in biopsies from women with cervical intraepithelial neoplasia. AB - Human papillomavirus antigen was found in 39 (16%) of 253 colposcopic biopsies from a group of women at high risk for cervical cancer who had been examined in the early 1970s. Immunohistochemical evidence of papillomavirus infection was found in 20 (30%) of 67 cervical intraepithelial neoplasia lesions infected with wart virus from these patients. When these results were compared with results of a similar study carried out in the early 1980s there was no significant difference in the prevalence of human papillomavirus antigen in cervical intraepithelial neoplasia lesions infected with wart virus from women who had been examined over a decade apart. PMID- 2993371 TI - Lymphoid and epithelial markers in small cell anaplastic thyroid tumours. AB - The expression of the lymphoid marker, common leucocyte antigen and of the epithelial marker, epithelial membrane antigen by small cell anaplastic thyroid tumours was studied in 53 tumours, using the peroxidase--antiperoxidase and avidin--biotin techniques. Common leucocyte antigen was found in 33 of the 53 tumours; six tumours were positive for epithelial membrane antigen; and the remaining 14 tumours were negative for both markers. These results support the suggestion that most small cell anaplastic thyroid tumours are of lymphoid origin. Survival data for patients with tumours positive for common leucocyte antigen showed a significantly better prognosis than did the data for patients with small cell tumours that did not express this marker (p less than 0.02). PMID- 2993373 TI - Biphasic plasma aldosterone responses to four single-dose ACTH regimens. AB - Adrenocorticotropic hormone (ACTH) administration increases cortisol synthesis but produces a biphasic aldosterone response. Some investigators believe that the hypercortisolism from prolonged ACTH administration is responsible for this aldosterone response. The present study evaluated the plasma aldosterone response to four acute single-dose ACTH regimens that produced only a transient increase in plasma cortisol. Fourteen normal adult men received (1) 1-18 ACTH intravenous bolus (IV), (2) 1-18 ACTH intramuscular (IM), (3) 1-39 ACTH (IM), and (4) 1-24 ACTH (IV). The plasma aldosterone increased within one hour and tended to parallel the cortisol increment with all four ACTH regimens. With all of these ACTH regimens, the plasma aldosterone level decreased below placebo with 1-24 ACTH (IV) (24 hours), 1-39 ACTH (IM) (24 hours), and 1-18 ACTH (IV) and (IM) (48 hours) at a time when the cortisol had returned to normal. These results suggest that the delayed ACTH-induced aldosterone inhibition production are not directly related to cortisol production and do not require prolonged ACTH administration. These observations are consistent with ACTH induction of a nonaldosterone mineralocorticoid, which is independently suppressing aldosterone production. PMID- 2993372 TI - Intracytoplasmic inclusions in B prolymphocytic leukaemia: ultrastructural, cytochemical, and immunological studies. AB - The intracytoplasmic inclusions seen in most cells from a patient with B prolymphocytic leukaemia were analysed using both light and electron microscopy. They consisted of a dense homogeneous structure and were surrounded by a membrane, which had no continuity with the Golgi cisternae or the endoplasmic reticulum; some inclusions had a clear association with small lysosomal granules. Immunofluorescence and immunoperoxidase studies using light microscopy failed to elucidate completely the nature of the inclusions, but immunocytochemical reactions performed using electron microscopy suggested an immunoglobulin nature. All inclusions were negative for acid phosphatase and periodic acid Schiff. The nature of the inclusions described in the prolymphocytes of this patient were compared with those previously recorded in B prolymphocytic leukaemia. PMID- 2993374 TI - Morphology of central terminations of intra-axonally stained, large, myelinated primary afferent fibers from facial skin in the rat. AB - Horseradish peroxidase was intra-axonally injected into functionally identified primary afferent fibers within the rat spinal trigeminal tract in order to study the morphology of their central terminations. They were physiologically determined to be large, myelinated, cutaneous primary afferents by means of electrical and mechanical stimulation of their receptive fields. Ninety-three axons that innervated vibrissa follicles, guard hair follicles, and slowly adapting receptors were stained for distances of 4-12 mm at the levels of the main sensory nucleus, spinal trigeminal nucleus, and rostral cervical spinal cord. The collaterals of single axons from these receptors formed terminal arbors in the outer part of the spinal trigeminal nucleus rostral to and near the level of the obex (rostral type collaterals). In the rostral part of the subnucleus caudalis (Vc) they were confined to lamina V (caudalis type collaterals) and in the caudal part of Vc and in cervical segments they were confined to lamina III/IV (spinal-dorsal-horn-type collaterals). There were no transitional forms between the rostral and caudalis types, but there was a transitional form between the caudalis and spinal dorsal horn types. This transitional form was distributed in laminae III/IV and V. The terminal arbors of the rostral type of collaterals formed an interrupted, rostrocaudally oriented column like those seen in the lumbar dorsal horn, but the column shifted down to lamina V near the obex, and more caudally, gradually shifted upward to lamina III. Major morphological differences were not observed among the three different functional types of collaterals with respect to the rostrocaudal distribution of collaterals, and the shape and location of collaterals. The differential laminar distribution of collateral arbors of single axons along the rostrocaudal axis distinguishes the spinal trigeminal nucleus from the spinal dorsal horn where functional types of mechanoreceptive afferents form continuous or interrupted sagittal columns of terminal arbors that do not shift dorsoventrally within segments. PMID- 2993375 TI - Coliform numbers in the stomach and small intestine of healthy pigs following weaning at three weeks of age. AB - In 60 weaned and 52 unweaned pigs, aged between 21 and 32 days, the number of coliform organisms in the gastric contents was found to be related to their pH value. The contents were generally more acidic in the weaned animals and contained fewer coliforms. Haemolytic coliform organisms were never identified among selected isolates examined from the gastric contents although in weaned animals the haemolytic enterotoxigenic Escherichia coli serotype 0149: K91, K88a,c (Abbotstown strain) was commonly recovered from the intestines. After weaning, this strain appeared to colonize the anterior small intestine from lower down the tract, and its presence was associated with an increase in the total coliform count at the site from which it was isolated. Rotaviruses were also more commonly detected in the intestinal contents of weaned than unweaned pigs, but their appearance after weaning appeared to be preceded by the proliferation of haemolytic E. coli. The presence of either or both of these potentially enteropathogenic organisms never induced diarrhoea, and although faecal water content increased significantly after weaning, this occurred independently of the presence of both agents. PMID- 2993376 TI - Generalized parvovirus infection with inclusion-body myocarditis in two kittens. AB - Two 13-day-old kittens showed a disease characterized by degeneration, inflammatory response and inclusion bodies in intestine, liver and in one case the myocardium. Inclusions contained typical parvovirus particles. Either a generalized parvovirus infection by the causal agent of feline panleukopenia or an infection due to an unknown parvovirus is considered a possible cause of the disease. A possible relationship of the agent to canine parvovirus is also considered. PMID- 2993377 TI - Rheumatoid factors in natural retrovirus infections of the cat and cattle. AB - The occurrence of IgM-rheumatoid factors (RFs) was investigated in natural retrovirus infection of cats with feline leukaemia virus and of cattle with bovine leukaemia virus. IgM-RFs of polyclonal character were detected. No significant differences were observed between the amounts of IgM-RFs in the retrovirus-infected animals and their respective controls. PMID- 2993378 TI - Chlorpromazine-induced lupuslike disease. Case report and review of the literature. AB - A lupuslike disease with cutaneous manifestations secondary to the use of chlorpromazine (Thorazine) is presented. Skin biopsy specimens demonstrated classic findings consistent with lupus erythematosus and abundant mucin deposition in the cutaneous lesions--a finding not previously reported to occur in drug-induced lupuslike disease. Laboratory and serologic examinations included a positive antinuclear antibody titer, presence of single-stranded deoxyribonucleic acid (DNA), absence of double-stranded DNA, presence of antihistone antibodies, normal complement level, increased IgM level, and prolongation of partial thromboplastin time. These data confirmed our clinical and histologic diagnosis. Cessation of the medication (chlorpromazine) led to resolution of the skin abnormalities. Electron microscopy revealed electron-dense drug metabolites in vascular endothelial cells, as well as tubuloreticular inclusion bodies. This is only the second reported case of chlorpromazine-induced lupuslike disease with cutaneous manifestations. PMID- 2993379 TI - Genital Paget's disease and urinary tract malignancy. AB - Eight cases of Paget's disease of genital mucosa with malignancy of the lower urinary tract are described. In five it was apparent that there was concurrence of two separate malignancies. In one patient with long-standing Paget's disease of the genital mucosa, carcinoma of the urethra and bladder developed subsequently in continuity with the genital lesion; the two lesions were indistinguishable histologically, suggesting extension of the Paget's disease into urothelium. In two patients with bladder malignancy, there was histologic evidence of outward pagetoid extension of this process along urothelium and onto the genital mucosa. The significance of genital Paget's disease is discussed in the light of these findings, and the possible origins of Paget cells within the epidermis are reviewed. PMID- 2993380 TI - Metastatic cutaneous carcinoid. AB - A case of malignant bronchial carcinoid with multiple cutaneous metastases is reported. Despite the extremely aggressive behavior of the disease and the presence of multiple hepatic metastases, the patient did not exhibit the usual clinical features of the carcinoid syndrome. The relevance of the finding of elevated 24-hour urinary vanillylmandelic acid levels in patients with carcinoid tumors is discussed. PMID- 2993381 TI - Enhancement of cervical intraspinal tumors in MR imaging with intravenous gadolinium-DTPA. AB - Intravenous Gd-diethylenetriamine pentaacetic acid (Gd-DTPA) was administered to 10 successive patients with cervical intraspinal tumors. Contrast enhancement was seen in each case. Gadolinium-DTPA was useful in outlining the tumoral masses, as well as in distinguishing intramedullary from extramedullary neoplasms. PMID- 2993382 TI - Distinguishing neuroblastoma from Wilms tumor by computed tomography. AB - The CT appearances of 18 clinically proven cases of abdominal neuroblastoma were reviewed retrospectively with the aim of characterizing those features that best distinguish them from Wilms tumors. The study found that the most common characteristics of neuroblastoma on CT are usually a suprarenal mass displacing the kidney and either growing down into the hilum or actually infiltrating the kidney; extension of the tumor across the midline to displace the major vessels; presence of calcification in the tumor. PMID- 2993383 TI - Heritability of forskolin and hormone-stimulated adenylate cyclase activity in human lymphocytes. AB - Isoproterenol, prostaglandin E1 and forskolin-stimulated cyclic AMP accumulation were compared in intact lymphocytes obtained from nine monozygotic and nine sib pairs matched for age and sex. Heritability was calculated by three different methods, two based on the intraclass correlation coefficients and one based directly on the variances. Only for forskolin is a significant proportion of variance (0.68-0.91) attributable to genetic factors, suggesting that forskolin stimulated activity may prove to be a valuable genetic marker in studies of human pathology. Neither basal nor isoproterenol and prostaglandin E1-stimulated activity show significant heritability in intact human lymphocytes. The individual differences observed in levels of beta-adrenergic and prostaglandin stimulated receptor activity in human lymphocytes are, therefore, most likely due to environmental factors. PMID- 2993386 TI - Effects of alloxan on the islets of Langerhans: stimulation and inhibition of cyclic AMP production. AB - Alloxan exerts a selective impairment of the insulin-producing B-cells of the islets of Langerhans, which may result in diabetes mellitus. The effects of alloxan on cyclic AMP metabolism in isolated mouse islets of Langerhans were investigated. Alloxan caused an immediate increase in islet content of cyclic AMP, whereas a subsequent glucose-stimulated increase of islet cyclic AMP content was inhibited in alloxan-exposed islets. No corresponding effects of the drug were, however, found on either islet adenylate cyclase or cyclic AMP phosphodiesterase activities in broken cell preparations. It appears unlikely that there is a direct interaction between alloxan and the enzyme molecules leading to irreversible changes. Alloxan may rather affect some metabolic factor essential for optimal enzyme function. The inhibition of glucose-stimulated increase in islet ATP content and adenylate energy charge in alloxan-treated islets suggests that such a factor might be dependent on intact ATP generation. PMID- 2993385 TI - Evidence for two specific affinity states of 3H-antagonist binding to cardiac beta-adrenergic receptors and influence of Gpp(NH)p. AB - Binding of the lipophilic antagonist ligand 3H-DHA and the hydrophilic antagonist ligand 3H-CGP 12177 to beta-adrenergic receptors of rat heart ventricular membranes was studied. Quantitative analysis of the binding data indicated the existence of two specific affinity states of the beta-adrenergic receptor population aside from a third non-specific binding site for 3H-DHA. In order to exclude that the biphasic saturation isotherm may be due to retained endogenous agonist, crude membranes as well as modified membranes were used. In the latter a 99% reduction of noradrenaline concentration was obtained by washing and preincubation or by catecholamine depletion. Two affinity states of antagonist binding could be demonstrated independently from the kind of membrane suspension. A biphasic dissociation of 3H-DHA by unlabelled (-)-alprenolol was also found in kinetic studies. In crude or washed membranes of untreated rats the guanine nucleotide Gpp(NH)p affected saturation and antagonist competition curves. However, this was not observed in catecholamine-depleted membranes of reserpine treated rats. Stereoselectivity of the high and the low affinity state was demonstrated in competition experiments with (-) - and (+) -alprenolol in catecholamine-depleted membranes. The data are best explained by assuming a ternary complex model (1) in which antagonists, instead of passively occupying binding sites, play an active role in receptor mechanisms. Based on this model, it is assumed that beta-adrenergic antagonists bind with high affinity to the free form of the receptor and with low affinity to the precoupled form. Furthermore, an interaction of Gpp(NH)p with the regulatory component is proposed. PMID- 2993387 TI - Inhibition of cyclic AMP-dependent protein kinase-induced changes in the kinetic properties of hepatic pyruvate kinase by the specific cyclic AMP antagonist, the (Rp)-diastereomer of adenosine cyclic 3',5'-phosphorothioate. AB - The effects of the diastereomers of adenosine cyclic 3',5'-phosphorothioate, (Sp) and (Rp)-cAMPS, on the kinetic properties of pyruvate kinase were studied in hepatocytes isolated from fed rats. Incubation of the cells with the cAMP dependent protein kinase agonist, (Sp)-cAMPS, produced a concentration-dependent increase in S0.5 for phosphoenolpyruvate, but had no effect on Vmax. The (Sp) cAMPS-treated enzyme was more sensitive to inhibition by alanine and ATP and, under the same conditions, was less responsive to activation by fructose-1,6 bisphosphate when assayed at a subsaturating phosphoenolpyruvate concentration. Incubation of the hepatocytes with only the cAMP-dependent protein kinase antagonist, (Rp)-cAMPS, produced no change in any kinetic parameters, but did suppress the (Sp)-cAMPS- or glucagon-induced increase in the S0.5 for phosphoenolpyruvate with IC50 values of 10 microM and 5 microM (Rp)-cAMPS. (Rp) cAMPS is exerting an effect on the kinetic properties of pyruvate kinase through inhibition of cAMP-dependent protein kinase. PMID- 2993384 TI - Opiate and dopamine stimulate different GTPase in striatum: evidence for distinct modulatory mechanisms of adenylate cyclase. AB - Previous studies demonstrated that opiate inhibition of adenylate cyclase (AC) in striatal membranes is related to an opiate-stimulated GTPase with a low Km. Dopamine (DA) also dose-dependently activates a high affinity GTPase, with a pattern of stimulation and a receptor selectivity (D1 type) similar to those observed in DA activation of striatal AC. Moreover, the DA- and the opiate sensitive GTPase activities have different sensitivities to agents that affect the inhibition of AC, such as Na+ and N-ethylmaleimide (NEM), or the stimulation, such as cholera toxin (CTX). Thus, the impairment of opiate-dependent inhibition of AC in the absence of Na+ ions or after NEM pretreatment of the membranes is parallel with preferential impairment of the opiate-dependent GTPase. On the contrary, selective blocking by CTX of the DA-dependent GTPase leads to the enhancement of AC stimulation by DA. These results suggest that DA activation of striatal AC is related to a GTPase that is specifically stimulated by DA and is associated with the Ns protein. A distinct Ni protein seems to be responsible for the opiate effect on AC and GTPase. PMID- 2993388 TI - Adipocyte cyclic nucleotide phosphodiesterase activation by vanadate. AB - The addition of vanadate (Na3VO4) to intact isolated rat adipocytes stimulated cAMP phosphodiesterase activity (Type IV) in the particulate (P2) fraction. Vanadate increased the Vmax of the Type IV phosphodiesterase activity without affecting its apparent substrate affinity. Na3VO4 also stimulated cAMP hydrolysis of cell-free particulate and cytosolic fractions, but this activation required the presence of reduced glutathione (GSH). The mixture of vanadate and glutathione appeared as an emerald green solution (V-GSH complex), which was shown by EPR to contain vanadyl ion. No effect of either GSH or Na3VO4 alone on cell-free particulate cAMP phosphodiesterase activity was observed; however, Na3VO4, alone or in combination with GSH, stimulated cGMP hydrolysis in this subcellular fraction. The V-GSH complex increased the Vmax of the particulate cAMP phosphodiesterase activity without affecting its apparent Km. The activating effect of the complex was rapid in onset, persistent over 30 minutes, and reversible. The EC50 for activation of the particulate cAMP phosphodiesterase was approximately 5 microM Na3VO4 (maintaining the GSH:Na3VO4 molar ratio at 2:1); maximal stimulation was achieved at 0.1 mM Na3VO4. Purified microsomal membranes showed activation similar to that of the P2 fraction, while only a 60% stimulation was observed in purified plasma membranes. The V-GSH complex increased basal insulin-activated Type IV phosphodiesterase activity to a common maximal level. Detergent-solubilized cAMP-phosphodiesterase from the P2 fraction was stimulated 2.5-fold by the V-GSH complex. Limited trypsin treatment of P2 membranes activated cAMP phosphodiesterase and abolished the stimulatory effect of the V-GSH complex. These results are generally consistent with the hypothesis that V-GSH complex activates Type IV phosphodiesterase by an indirect mechanism, which appears to involve predominantly membrane bound components that may be biologically important enzyme regulatory elements. PMID- 2993389 TI - Effect of ACTH and oxytocin treatment on lactoferrin and citrate in cows' milk. AB - Six dairy cows were treated before milkings with either oxytocin (Pitocin, 20 i.u.) or ACTH (Synacthen, 150 i.u.), principally to determine their effect on the ratio of citrate: lactoferrin concentrations in the milk. With ACTH treatment, after 3 d milk yield and citrate concentration decreased significantly, lactoferrin and bovine serum albumin (BSA) concentrations increased significantly. Somatic cell counts (SCC) increased temporarily in the milk of three of the cows which previously had greater than 100 000 cells/ml. Lactoferrin yield remained fairly constant but citrate yield was significantly reduced. The citrate: lactoferrin molar ratio decreased from 1373 to 606. With oxytocin treatment, after 4 d milk yield first increased and then significantly decreased, citrate concentration decreased significantly while there were no significant changes in lactoferrin or BSA concentration or in the yield of any other milk constituents. The citrate: lactoferrin molar ratio decreased from 1621 to 1301. There were no significant changes in SCC either during treatment or 4 d after treatment but there was a significant rise at 16 d after treatment. It was concluded that in lactating cows both hormones affected citrate and lactoferrin concentrations in the direction that would improve the antibacterial properties of milk, but that this was accompanied by adverse effects on milk secretion. The extent of the change was not sufficient to be likely to produce inhibition of coliform bacteria. PMID- 2993391 TI - Response in two commercial Holstein herds to addition of sodium bicarbonate to alfalfa hay-based diets. AB - Feeding trials were conducted in two commercial dairy herds to evaluate the addition of .8% sodium bicarbonate to alfalfa hay-based diets. Approximately half of each herd served as controls and the other half was fed the same diet with sodium bicarbonate. A total of 1280 Dairy Herd Improvement Association lactation records were obtained in the two herds during the trials. Cows in herd 1 were milked three times daily and cows in herd 2 were milked twice daily. In herd 1, milk production from control and bicarbonate groups was: first lactation cows, 7491 and 7748 kg/cow; second lactation cows, 8363 and 8791 kg/cow; and third and higher lactation cows, 8713 and 9562 kg/cow. There were no differences in milkfat or solids-not-fat percentages between treatment groups. In herd 2, milk production from control and bicarbonate groups was: first lactation cows, 6800 and 7158 kg/cow; second lactation cows, 8487 and 8082 kg/cow; and third and higher lactation cows, 8807 and 8216 kg/cow. First lactation cows fed sodium bicarbonate had a lower milk fat percentage than controls. There were no other differences in milk fat or solids-not-fat percentages between treatment groups. PMID- 2993390 TI - Addition of buffers to high quality alfalfa hay-based diets for dairy cows in early lactation. AB - Beginning 4 wk postpartum, 14 Holstein cows were paired according to expected calving date and lactation number and assigned randomly to one of two treatments in a single reversal experiment designed to study the effects of added buffers to high quality alfalfa hay-based diets. Cows were offered daily 14.5 kg of concentrate containing 0 or 2% sodium bicarbonate and .5% magnesium oxide (as fed), plus free choice high quality alfalfa hay in a ratio of forage:grain approaching 50:50 (dry matter). No differences were in milk production, milk fat percentage, fat-corrected milk, or dry matter intake. Dry matter intake of grain was decreased with addition of buffers. Kilograms fat-corrected milk per kilogram of dry matter intake were 1.41 and 1.45 for control and buffered diets. No differences were significant for rumen pH, ammonia concentration, molar proportions of acetate and propionate, or ratio of acetate: propionate. Concentrations of total volatile fatty acids were higher for cows fed the buffered diet. No differences were in blood acid-base status or in various serum metabolites. Urine pH was 8.11 and 8.20 for control and buffered diets. Fractional urinary excretion of magnesium and sodium was greater when cows consumed the buffered diet. Diet digestibilities and rate of passage were not affected by dietary buffers. Addition of sodium bicarbonate plus magnesium oxide did not improve performance of early lactation cows fed high quality alfalfa hay as the sole forage. PMID- 2993392 TI - The use of intracellular techniques in the study of the cochlear nucleus. AB - This paper describes part of the anatomy and physiology of the cochlear nucleus and some of the recent techniques developed to study the structure and function of the cochlear nucleus. The cochlear nucleus is the first region in the brain stem that receives information from the auditory nerve. The use of intracellular markers has made possible a new detail of description that should increase significantly our understanding of how the cochlear nucleus organizes the neural information transmitted to it from the auditory nerve. PMID- 2993393 TI - Use of brain slices in the study of the auditory system: spatial and temporal summation of synaptic inputs in cells in the anteroventral cochlear nucleus of the mouse. AB - One of the more dramatic technological developments in recent years in neurobiology is the ability to extract a slice of brain tissue and to maintain its viability over extended periods of time. The development of brain slice preparations of the mammalian central nervous system provides a powerful tool for studying the physiology and pharmacology of neurons. In vitro preparations allow stable intracellular recordings to be made from cells. The characteristics of synaptic potentials and the intrinsic electrical properties can be measured while the extracellular environment is controlled and manipulated. Intracellular physiological studies in the anteroventral cochlear nucleus reveal that different morphological classes of cells have characteristic intrinsic electrical properties. Bushy cells are particularly well suited to preserve and convey the temporal firing pattern of inputs from the auditory nerve. Stellate cells can transform inputs from the auditory nerve by summing in time and space. PMID- 2993394 TI - The role of suppression in psychophysical measures of frequency selectivity. AB - The neural output of the cochlea consists of a structured spectral representation of the input waveform. Psychophysical studies of frequency selectivity provide measures of the quality of this spectral representation based on the detection of a spectrally narrow signal in the presence of competing acoustic inputs. Results are often interpreted in terms of the underlying physiological processes. The present paper begins with a summary of the basic principles and data and a review of recent efforts to extend the theoretical framework to account for a wider range of phenomena. It then discusses and evaluates specific assumptions concerning the role of suppression in psychophysical measures of frequency selectivity, an area of research that has received much attention in recent years. The goal is to provide a summary of current answers to basic questions at a time of rapidly expanding knowledge about both the physiology and psychophysics of frequency selectivity. PMID- 2993395 TI - Nuclear magnetic resonance spectroscopy: its evolving role in the study of myocardial metabolism. PMID- 2993396 TI - Effect of the renin-angiotensin system on limb circulation and metabolism during exercise in patients with heart failure. AB - The maximal aerobic exercise capacity of patients with chronic heart failure is frequently decreased because of inadequate blood flow to working skeletal muscle. To investigate whether this reduced flow is in part due to interference by angiotensin II with arteriolar dilation in working muscle, the effect of the angiotensin-converting enzyme inhibitor captopril on leg blood flow, leg vascular resistance, leg oxygen consumption (VO2) and leg lactate release during maximal upright bicycle exercise was examined in 12 patients with heart failure (maximal VO2 10.7 +/- 3.1 ml/min per kg). Captopril decreased leg resistance at rest (258 +/- 115 to 173 +/- 67 U, p less than 0.01) and maximal exercise (68 +/- 69 to 45 +/- 29 U, p less than 0.01) associated with proportionately similar decreases in systemic vascular resistance. However, maximal exercise duration and maximal VO2 were unchanged and, at identical peak exercise work times, there was no improvement in leg blood flow (2.0 +/- 0.9 to 2.0 +/- 1.1 liters/min, p = NS), leg VO2 (261 +/- 104 to 281 +/- 157 ml/min, p = NS) or leg lactate release (269 +/- 149 to 227 +/- 151 mg/min, p = NS). These data suggest that, during exercise in patients with heart failure, angiotensin II does not interfere with blood flow to working skeletal muscle. PMID- 2993397 TI - Hemodynamic and clinical significance of the pulmonary vascular response to long term captopril therapy in patients with severe chronic heart failure. AB - Exercise capacity in patients with left heart failure is closely related to the performance of the right ventricle and the pulmonary circulation. To determine the significance of changes in pulmonary resistance during long-term vasodilator therapy, hemodynamic studies were performed before and after 1 to 3 months of treatment with captopril in 75 patients with severe chronic left heart failure. Patients were grouped according to the relative changes in pulmonary and systemic resistances during long-term therapy: patients in Group I (n = 24) showed greater decreases in pulmonary arteriolar resistance (PAR) than in systemic vascular resistance (SVR) (% delta PAR/% delta SVR greater than 1.0), whereas patients in Group II showed predominant systemic vasodilation (% delta PAR/% delta SVR less than 1.0). Despite similar changes in systemic resistance, patients in Group I showed greater increases in cardiac index, stroke volume index and left ventricular stroke work index (p less than 0.01 to 0.001) but less dramatic decreases in mean systemic arterial pressure (p less than 0.02) than did patients in Group II. Despite similar changes in left ventricular filling pressure, patients in Group I showed greater decreases in mean pulmonary artery and mean right atrial pressures (p less than 0.02 to 0.01) than did patients in Group II. Pretreatment variables in Groups I and II were similar, except that plasma renin activity was higher (8.7 +/- 2.1 versus 3.0 +/- 0.6 ng/ml per h) and serum sodium concentration was lower (133.1 +/- 0.9 versus 137.1 +/- 0.6 mEq/liter) in Group II than in Group I (both p less than 0.05). Both groups improved clinically after 1 to 3 months, but symptomatic hypotension occurred more frequently in Group II than in Group I (36 versus 8%) (p less than 0.005). These findings indicate that changes in the pulmonary circulation modulate alterations in both right and left ventricular performance during the treatment of patients with left heart failure. Hyponatremic patients are likely to experience symptomatic hypotension with captopril because they are limited in their ability to increase cardiac output as a result of an inadequate pulmonary vasodilator response to the drug. PMID- 2993398 TI - Adherence to high-carbohydrate, high-fiber diets: long-term studies of non-obese diabetic men. AB - High-carbohydrate, high-fiber (HCF) diets provide short-term benefits for selected individuals with diabetes. Long-term effects of HCF diets, however, have not previously been documented. The effectiveness and acceptability of HCF diets for 14 men with diabetes followed for 4 years as outpatients were assessed. Hospital HCF diets with 70% of energy as carbohydrate and 65 gm plant fiber daily lowered insulin doses, serum cholesterol, and triglycerides. Home HCF diets with 55% to 60% carbohydrate and 50 gm plant fiber daily sustained improvements throughout the observed period of up to 86 months. Of the 11 patients for whom diet adherence was evaluated, 9 had good to excellent adherence, 1 had fair adherence, and 1 had poor adherence. PMID- 2993399 TI - Dietary fiber constituents of selected fruits and vegetables. AB - This study compared the dietary fiber (DF), neutral detergent fiber (NDF), cellulose, hemicellulose, lignin, and pectin content of selected fruits and vegetables. Apples and peaches (fresh and canned), oranges (fresh), strawberries (fresh, canned, and frozen), carrots, green beans, and potatoes (fresh, fresh cooked, canned, and frozen), and tomatoes (fresh, fresh cooked, and canned) were studied. When possible, two varieties, two stores, and name and store brands were chosen. Samples were analyzed for NDF, acid detergent fiber, 72% sulfuric acid, lignin, and pectin. From those values, DF, cellulose, and hemicellulose were calculated. Fresh fruits in gm/100gm wet weight had decreasing DF, NDF, and hemicellulose values as follows: apples, peaches, strawberries, and oranges. Apples were highest in cellulose; strawberries, highest in lignin; and oranges, highest in pectin. Fresh-cooked vegetables in gm/100gm wet weight have decreasing DF and NDF values as follows: green beans, carrots, potatoes, and tomatoes. Green beans were highest in cellulose and hemicellulose; potatoes highest in lignin; and carrots highest in pectin. On a wet-weight basis, fresh apples and peaches, fresh-cooked green beans, canned carrots, and canned and frozen potatoes were higher in DF and NDF than other forms of the fruit or vegetable. There were few differences according to stores, brands, or varieties of fruits and vegetables. On a dry-weight basis, fresh apples, peaches, strawberries, green beans, and tomatoes appear to have higher DF and NDF contents than their processed counterparts. Fresh-cooked carrots and fresh potatoes appear to have less DF and NDF than their canned and frozen counterparts. PMID- 2993400 TI - Dietary strategies in the treatment of reactive hypoglycemia. PMID- 2993401 TI - Cyanine and safranine dyes as membrane potential probes in cytochrome c oxidase reconstituted proteoliposomes. AB - Safranine and the cyanine dye, 3',3'-dipropylthiadicarbocyanine (diSC3-5), were examined as membrane potential probes in cytochrome c oxidase vesicles. The spectra of the vesicle-associated dyes resemble those of the same dyes in organic solvents, indicating that safranine and diSC3-5 probably dissolve in a hydrophobic region of the proteoliposomal membrane. This binding of safranine to proteoliposomes occurs with a dye-membrane dissociation constant in the micromolar range. The binding of safranine and of diSC3-5 to liposomes or proteoliposomes is accompanied by fluorescence enhancement. This enhanced fluorescence is quenched by respiration or by the establishment of a K+ diffusion potential by valinomycin (negative interior). An optimal dye/lipid ratio was required to secure maximum fluorescence quenching of the dyes, whether that quenching was active or passive. Calibrations of both the safranine and the diSC3 5 responses with K+ diffusion potentials were also affected by the dye/lipid ratio. At lower dye/lipid ratios, the calibration curve was linear at higher potentials but deviated from linearity at lower potentials. The converse was true at higher dye/lipid ratios. The non-linearity of the calibration curve at higher potential was attributed to a 'saturation' effect; it may also involve increased permeability of proteoliposomal membrane to protons. Destacking of dye at the lower dye/lipid ratio was probably responsible for the non-linearity of the calibration curves at lower potentials. When all these effects are taken into account, the steady-state value of delta psi generated during maximal proteoliposomal respiration was calculated to be between 140 and 160 mV (interior negative) when measured with either safranine or diSC3-5. We conclude that quantitative estimates of delta psi values can be made using these probes in cytochrome c oxidase reconstituted proteoliposomes provided that appropriate precautions are taken. PMID- 2993402 TI - An intracellular study of the synaptic input to sympathetic preganglionic neurones of the third thoracic segment of the cat. AB - In chloralose anaesthetized, paralyzed and artificially ventilated cats intracellular recordings were obtained from sympathetic preganglionic neurones (SPN) of the third thoracic segment of the spinal cord identified by antidromic stimulation of the white ramus T3. The synaptic input to SPNs was assessed, in cats with intact neuraxis or spinalized at C3, by electrical stimulation of segmental afferent fibres in intercostal nerves and white rami of adjacent thoracic segments and by stimulation of the ipsi- and contralateral dorsolateral funiculus and of the dorsal root entry zone of the cervical spinal cord. In both preparations SPNs showed on-going synaptic activity which predominantly consisted of excitatory post-synaptic potentials (EPSPs). Inhibitory post-synaptic potentials (IPSPs) were rarely observed. EPSPs were single step (5 mV) or, less frequently, large (up to 20 mV) summation EPSPs. The proportion of SPNs showing very low levels of on-going activity was markedly higher in spinal than in intact cats. Stimulation of somatic and sympathetic afferent fibres evoked early EPSPs (amplitude 3 mV, latency 5-22.3 ms), and late, summation EPSPs (amplitude up to 20 mV, latency 27-55 ms). Early and late EPSPs were evoked in nearly all SPNs in which this synaptic input was tested in the intact preparation (from 79-93% of the SPNs). In spinal cats, early EPSPs were evoked in 88% of the SPNs, whereas late EPSPs were recorded only in half of the neurones. No evidence for a monosynaptic pathway from these segmental afferent fibres to SPNs was obtained. In both intact and spinal cats, stimulation of the dorsolateral funiculus evoked early and late EPSPs in SPNs. Late EPSPs were recorded in 70% and 37% of the SPNs in intact and spinal cats, respectively. Early EPSPs, however, were evoked in all neurones. The early EPSPs evoked by stimulation of the dorsolateral funiculus had several components which are suggested to arise from stimulation of descending excitatory pathways with different conduction velocities. The following conduction velocities were calculated in intact (spinal) cats: 9.5-25 m/s (7.8 13.2 m/s), 5.7-9.5 m/s (5.5-7.8 m/s), 3.8-5.7 m/s (3.2-5.5 m/s), and 2.6-3.8 m/s (2.1-3.2 m/s). EPSPs of these various groups were elicited in a varying percentage in SPNs. EPSPs of the most rapidly conducting pathway were subthreshold for the generation of action potentials; some EPSPs of this group had a constant latency suggesting a monosynaptic pathway to SPNs. Stimulation of the dorsal root entry zone at the cervical level yielded essentially the same results as stimulation of the dorsolateral funiculus.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993403 TI - Modification of dobutamine- and terbutaline-induced calcium and fluid secretion from rat salivary glands by atenolol and butoxamine. AB - Saliva was elicited from rat salivary glands by terbutaline at i.p. doses of 1, 5, 10 and 25 mg/kg b.wt. but not by doses of 0.1 or 0.5 mg/kg b.wt. Dobutamine elicited no secretion at 1 or 2 mg/kg but did at 5, 10 and 25 mg/kg b.wt. At 5 mg/kg terbutaline evoked nearly maximal volumes but with dobutamine, volumes were small at this dosage. At dosages of 10 and 25 mg/kg volumes with the two agonists were similar for parotid, but with submandibular, the volumes evoked by dobutamine were nearly two times as high as those elicited by terbutaline. Mean [Ca] of parotid saliva was also similar at all dosages of dobutamine (approximately 12 mEq/liter) and generally similar at all dosages of terbutaline (11-15 mEq/liter). Mean [Ca] of dobutamine-elicited submandibular saliva was approximately 6, 7 and 8 mEq/liter at 5, 10 and 25 mg/kg b.wt, respectively. With parotid, [Ca] was approximately 10 mEq/liter at 1, 5 and 10 mg/kg b.wt. but increased to 16-18 mEq/liter at 25 mg/kg. The time course of calcium secretion is described for both agonists at each dosage. [Ca] of both glands was decreased 60 min after i.p. injection of 10 or 25 mg/kg doses of dobutamine or terbutaline but was not changed by 5 mg/kg doses. Administration of 10 mg/kg of atenolol, the selective beta 1 antagonist, 20 min prior to injection of a 10 mg/kg dose of either terbutaline (beta 2 agonist) or dobutamine (beta 1 agonist) blocked secretion from both glands, and prevented the usual agonist-induced reduction in glandular concentration of calcium. Butoxamine, on the other hand, did not modify effects of terbutaline on fluid secretion or depletion of glandular calcium; it did partially inhibit dobutamine-induced fluid and calcium secretion but not depletion of glandular calcium. The present data suggest that beta adrenoceptors of salivary glands are predominantly of the beta 1 subtype and that it is these that regulate calcium and fluid secretion. On the basis of the data with the antagonists, it is concluded that terbutaline activates beta 1 rather than beta 2 receptors since the beta 1 antagonist but not the beta 2 antagonist blocked secretory responses to terbutaline. PMID- 2993404 TI - The thyrotropin receptor and its role in Graves' disease. PMID- 2993405 TI - Metyrapone: an agent for melatonin as well as ACTH and cortisol secretion. AB - The metyrapone-test was used to study the influence of pituitary-adrenal activity on the pineal function in man. We recorded in blood the circadian rhythmicity of melatonin, ACTH and cortisol and the excretion of melatonin, cortisol and Porter Silber chromogens in urine before, during and after the administration of 750 mg metyrapone every 4 h during 24 h in two healthy volunteers and in two patients with asymptomatic, moderate hyperparathyroidism and a prolactin-secreting microadenoma, respectively. The present study confirmed our previous report on an increased excretion of melatonin per 24 h and per mmol creatinine during the administration of metyrapone. The excretory maximum preceded the maximal ACTH adrenal response. Serum melatonin remained unchanged during the administration of metyrapone. A second finding was depressed serum melatonin the night after the test in the subjects with the most marked ACTH-cortisol response following the metyrapone-test indicating suppression of melatonin secretion when ACTH-cortisol secretions were increased. A third finding was a late increase in diuresis appearing the day after metyrapone-administration while glomerular filtration rates did not show significant alterations. Thus, it is shown that the metyrapone induced cortisol inhibition stimulates both ACTH and melatonin secretion, while high ACTH and cortisol levels are accompanied by reduced S-melatonin levels. PMID- 2993406 TI - Postprandial hyperglycemia after ingestion of peeled and non-peeled fruits in type-2 diabetics. AB - Should diabetics prefer nonpeeled fruits in their diet? To answer this question 27 type-2 diabetics divided into three groups were examined on two different occasions under the same fasting conditions. The first group of patients received 300g of pears with peel and on another day 300g of peeled pears. The second group ate 300g of pears with peel and 230g of peeled pears (the 70g difference represents the weight of the peel). The third group of diabetics consumed 300g of apples with peel and 300g of apples without peel. Blood samples were collected before and 20, 40, 60, 80, 100, 120 and 140 min after fruit ingestion. No significant differences were noted in terms of mean blood glucose, serum insulin and serum triglyceride levels among the two meals (fruits with or without peel). This observation was confirmed in all groups studied. Peeled and nonpeeled fruits appear to produce the same hyperglycemia in type-2 diabetics, in spite of the high fiber content of the peel. Therefore, the suggestion of reducing postprandial hyperglycemia in diabetics by eating nonpeeled fruits does not seem to be justified. PMID- 2993407 TI - Effects of etomidate on cortisol biosynthesis in isolated guinea-pig adrenal cells: comparison with metyrapone. AB - The effects of the iv hypnotic etomidate on cortisol biosynthesis have been investigated in short term incubations of dispersed guinea-pig adrenal cells and were compared with those produced by metyrapone. Fifty percent inhibition of cortisol output was obtained at a final medium concentration of 3.5 10(-8) M (basal), 2.8 10(-8) M (ACTH-stimulated) for etomidate and of 5.10(-7) M (stimulated) for metyrapone. In the presence of etomidate, 11-deoxycortisol at 5.10(-8) M reached a peak value of 244 +/- 11% of control (mean +/- SE, n = 7). 17 alpha-Hydroxyprogesterone and progesterone were not significantly affected up to 10(-7) M, but at higher concentrations, all three precursors fell under their control values. Metyrapone induced a progressive rise of 11-deoxycortisol, from 10(-7) M upwards, to a maximum level at 10(-5) M (210 +/- 15% of control, mean +/ SE, n = 5). 17-Hydroxyprogesterone and progesterone concentrations were not significantly modified by metyrapone. The less active hypnotic L-enantiomer of etomidate had almost no inhibitory effect on cortisol production. The results obtained so far suggest that etomidate is a potent inhibitor of the mitochondrial cytochrome P-450 enzymes of the adrenal cortex, mainly the 11 beta-hydroxylase. At higher dose the cholesterol side-chain cleavage enzyme system seemed also to be affected. PMID- 2993408 TI - Anti-allergic action of 6-ethyl-3-(1H-tetrazol-5-YL) chromone (AA-344) in rats. I. Effect of AA-344 on passive cutaneous anaphylaxis in the hind paws of rats. PMID- 2993409 TI - [Real and false risks of local contraception: spermicides and the diaphragm]. AB - A critical analysis of recent publications about spermicides and their side effects has been made. The modern spermicides consist principally of nonoxynol-9 and benzalkonium chloride. The products are harmless and efficient when used correctly at each sexual intercourse. Experiments on rabbits and rats show that nonoxynol-9 is absorbed through the vaginal wall. Benzalkonium chloride is not absorbed, as shown by tests in women and rats. The diaphragm as a method of contraception does present several psychosomatic drawbacks. It appears harmless; allergy to rubber or the septic shock syndrome are extremely rare. PMID- 2993410 TI - The effects of 1,25-dihydroxyvitamin D3 on human T lymphocyte activation and proliferation: a cell cycle analysis. AB - Recent studies have demonstrated that 1,25-dihydroxyvitamin D3 (calcitriol), the most biologically active metabolite of vitamin D, is a potent inhibitor of both lectin- and antigen-driven human T lymphocyte proliferation. To better characterize this effect, we performed cell cycle analysis of both untreated and calcitriol-treated peripheral blood mononuclear cells after PHA stimulation. By using the metachromatic dye acridine orange and flow cytometry, we found that calcitriol blocks the transition from the early, low RNA compartment of G1 (G1A) to the late, higher RNA compartment of G1 (G1B). Consistent with this observation was the inability of exogenous IL 1 or phorbol myristic acetate to overcome calcitriol's suppression of DNA synthesis. Indomethacin slightly reversed calcitriol's inhibition of transition from early to late G1, suggesting a minor, prostaglandin-dependent component to calcitriol's antiproliferative activity. Finally, by using the monoclonal antibodies anti-Tac and OKT9, we found that calcitriol had no effect on IL 2 receptor expression, an early G1 event, but markedly inhibited transferrin receptor expression, an IL 2-dependent, late G1 event. Thus, analysis of calcitriol's effects on the expression of these T cell activation antigens provides further evidence of the cell cycle specificity of calcitriol's action in regulating human T lymphocyte proliferation. PMID- 2993411 TI - Surface marker characterization of EBV target cells in normal blood and tonsil B lymphocyte populations. AB - Human FACS-sorted B lymphocyte subpopulations were investigated for their susceptibility to immortalization by Epstein Barr virus (EBV). Only B cells reacting with the monoclonal antibody B2 were immortalized, whereas cells reacting with anti-human IgG or the monoclonal antibody BB2 were not responding. Cells positive or negative for IgM, IgD, Burkitt's lymphoma antigen (BLA), BB1, and HB2 were all transformed by EBV. PMID- 2993412 TI - The effects of vasoactive intestinal peptide on human natural killer cell function. AB - Vasoactive intestinal peptide (VIP) can be found at nerve endings in various tissues and has recently been shown to interact with human lymphocytes through an adenylate cyclase-linked receptor. Because various neuroendocrine factors are thought to influence immune responsiveness, we studied the effect of VIP on natural killer (NK) effector function. Human lymphocytes were incubated with 51Cr labeled K562 target cells in a 4-hr cytotoxicity assay in the absence or presence of increasing concentrations of VIP. As expected from its activation of adenylate cyclase, VIP was inhibitory at 10(-6) to 10(-10) M. Interestingly, however, when lymphocytes were preincubated with VIP for 30 or 60 min, then washed and added to target cells, a significant augmentation of NK activity ensued. Binding studies revealed that preincubation with VIP resulted in increased numbers of effector target conjugates, whereas cytotoxic activity in agarose was not affected at the single cell level. Studies with synthetic analogs of VIP revealed that the integrity of the 14-28 C-terminal amino acid sequence was essential for its activity in cytotoxicity. These data strongly suggest a functional role for VIP in modulating immune responses during neuroendocrine interactions with the immune system. PMID- 2993413 TI - Monoclonal antibodies to horse cytochrome c expressing four distinct idiotypes distribute among two sites on the native protein. AB - Antibody-secreting hybridoma cell lines produced from BALB/c mice that had been immunized with horse cytochrome c (cyt c) conjugated to hemocyanin yielded six hybridoma subclones that produced four monoclonal antibodies (mAb) with different patterns of cross-reactivity with a panel of evolutionarily variant cyt c. The recognition sites for three of these mAb lay in the same region of the intact molecule, because two of the mAb were sensitive to the amino acid residue present at sequence position 44, with one requiring threonine at position 47. The fourth mAb bound in another region of the molecule at a site that involves either residue 60 or residue 89. Synthetic peptides that included these residues did not react with these mAb, indicating that these sites may require interactions from noncontiguous regions of the molecule to bind antibody. The association constants for the interaction of the mAb with horse cyt c were very similar and of the order of 10(10) M-1. Specificity studies with anti-idiotypic sera and competition assays between mAb for binding to horse cyt c confirmed that the six positive hybridoma subclones produced from this fusion produced mAb that had one of these four distinct specificities. The idiotypes of these four mAb were serologically distinct, and were derived from Vh genes of the J558 family. PMID- 2993414 TI - Characterization of monoclonal antibodies to the beta-adrenergic antagonist alprenolol as models of the receptor binding site. AB - Antibodies to receptor ligands have been valuable in understanding the nature of receptor-ligand interactions. We have developed four monoclonal antibodies to the beta-adrenergic receptor antagonist alprenolol by immunizing A/J mice with (-) alprenolol coupled to keyhole limpet hemocyanin. The antisera from these mice displayed specific [3H]dihydroalprenolol ([3H]DHA) binding that was inhibited by alprenolol, propranolol, and isoproterenol. Somatic cell fusion of spleen cells from the immunized mice to SP2/0 myeloma cells, followed by limited dilution subcloning, resulted in the isolation of four hybridomas (1B7, 5B7, 5D9, and 2G9) demonstrating three different classes of ligand binding characteristics. 1B7 had the highest binding affinity for antagonists based on Scatchard analysis (Kd [125I]- CYP = 1.4 X 10(-10) M; Kd [3H]DHA = 6.5 X 10(-9) M), and was the only antibody to demonstrate agonist-inhibition of [3H]DHA binding. Ki values computed from competitive inhibition curves of [3H]DHA binding to 1B7 resulted in a rank order of potency similar to that of beta-2-adrenergic receptors: (-)-propranolol greater than acebutolol amine greater than isoproterenol greater than (+) propranolol greater than epinephrine greater than norepinephrine. 5B7 and 5D9 exemplified a second class of antibody. This pair had lower antagonist binding affinities (Kd [3H]DHA = 2 X 10(-8) M and 2.5 X 10(-7) M, respectively) and was stereoselective in binding receptor antagonists: (-)-propranolol greater than (+) propranolol greater than acebutolol amine. Agonist inhibition of [3H]DHA binding to these antibodies could only be observed at very high concentrations (greater than 10(-4) M agonist), and was not dose-dependent. Finally, the class of anti alprenolol monoclonal antibodies represented by 2G9 had the lowest antagonist binding affinity of all (IC50 alprenolol = 1 X 10(-5) M), did not demonstrate ligand stereoselectivity, and did not recognize agonists. We propose that antibodies raised against beta-adrenergic receptor ligands demonstrating stereoselective agonist binding will also demonstrate high affinity antagonist binding, and that they will closely parallel the binding characteristics of the receptor. According to this "agonist best-fit hypothesis," anti-idiotypic antibodies raised against the binding site of these idiotypes might contain true mirror images of the beta-adrenergic receptor binding site. PMID- 2993415 TI - Immunosuppressive effects of benzidine in mice: evidence of alterations in arachidonic acid metabolism. AB - Benzidine (4,4'-diaminobiphenyl), a known human bladder carcinogen used in the synthesis of dyes, was immunosuppressive in mice after subchronic exposure. Suppression, particularly of cell-mediated immunity, occurred at dose levels previously found to be subtumorigenic in mice, as evidenced by suppressed lymphoproliferative and delayed hypersensitivity responses. In addition, benzidine exposure was found to decrease host resistance, including resistance to the growth of transplantable tumor cells and infection with Listeria. These data suggest that the development of neoplastic disease may be facilitated by the ability of benzidine to alter the immune response. The mechanism(s) responsible for immunosuppression by benzidine, however, is probably not the same as that responsible for its direct carcinogenicity. The addition of benzidine in vitro to mitogen-activated lymphocytes mimicked the suppression of lymphocyte responsiveness in vivo. In vitro studies suggested that alterations in metabolites of the arachidonic acid/lipoxygenase pathway were responsible for the immune alterations. Benzidine and the lipoxygenase inhibitor NDGA inhibited arachidonic acid metabolism and the mitogen response in lymphocytes, whereas the cyclooxygenase inhibitor indomethacin was ineffective. Addition of 8brcGMP partially restored benzidine-suppressed responses, whereas arachidonic acid potentiated the suppression. These data are consistent with the hypothesis that alterations in lymphocyte functions may occur as a result of quantitative changes or depletion of conversion products in the arachidonate/lipoxygenase pathway induced by the addition of compounds that serve as co-oxidative substrates for hydroperoxidases, the prototype being benzidine. PMID- 2993416 TI - Stimulation of a cell-mediated cytotoxic response to FeLV-induced T cell lymphomas in the cat. AB - Nonspecific cell-mediated cytotoxicity was examined in the peripheral blood and spleens of normal and vaccinia virus-infected adult domestic cats. Natural cytotoxic (NC)-like cells, as measured by lysis of vaccinia- or HSV-infected, adherent cat tongue cells, were found in both the spleen and peripheral blood of normal, nonimmune cats. Cytotoxicity was expressed in a 16-hr assay but not in a 4-hr assay. Natural killer (NK)-like cells, as measured by lysis of an FeLV induced lymphoid tumor cell line (FL-74) growing in suspension, were found in the spleen but not PBL, and required a 16-hr assay for expression. Infection with vaccinia virus did not increase the activity of feline NC-like cells in either the peripheral blood or the spleen. NK-like function, however, was increased. Cytotoxicity peaked 6 days post-infection and required a 16-hr assay for maximal expression of cell lysis. Furthermore, a cell with cytotoxic characteristics of the spleen NK-like cell appeared at low levels in the circulation at 6 days post vaccinia infection. NK-like cells from vaccinia-infected cats showed some cytotoxicity for FL74 targets in a 4-hr assay. The cat thus possesses at least two functionally different populations of naturally cytotoxic cells. NC-like cells are found in the spleen and peripheral blood, lyse virus-infected monolayer targets, and are not activated by infection. NK-like cells are found in the spleen, lyse-lymphoid tumor targets, and can be activated by infection, with their peak activity occurring 6 days after infection. PMID- 2993417 TI - Molecular cloning and DNA sequence analysis of genes encoding cytotoxic T lymphocyte-defined HLA-A3 subtypes: the E1 subtype. AB - Influenza-specific cytotoxic T cells restricted by HLA-A3 and allogeneic CTL specific for HLA-A3 recognize differences between serologically indistinguishable HLA-A3 antigens. Previous biochemical studies have indicated that such differential recognition can be explained by alterations in the primary structure of class I heavy chains. Characterization of these sequence differences may therefore identify portions of the class I molecule that form determinants recognized by CTL. In this study, we describe the cloning and sequencing of an HLA-A3 subtype from donor E1 (E1-A3). Cloning of the gene encoding E1-A3 was simplified by determining that a 15.5-kb BamHI fragment contains the complete gene and is characteristic of HLA-A3 and only one other class I gene (HLA-A11). Comparison of the E1-A3 sequence to that of a previously sequenced HLA-A3 gene for exons encoding extracellular class I domains revealed three nucleotide differences. All of these differences were located within a discrete region of exon 3 (encoding the alpha 2 domain) and result in a change of two amino acids, at positions 152 (Glu----Val) and 156 (Leu----Gln). This finding suggests that these amino acids are crucial for the information of a determinant recognized by CTL. Furthermore, the altered nucleotide sequence of E1-A3 is identical to the sequence of the HLA-Aw24 gene for codons 128 to 161. These observations of multiple clustered changes in the E1-A3 subtype (relative to the prototype sequence) and identity of the altered sequence with the sequence of another class I gene support the concept that gene conversion is a primary mechanism for the generation of class I polymorphism. PMID- 2993418 TI - In vivo administration of purified human interleukin 2. II. Half life, immunologic effects, and expansion of peripheral lymphoid cells in vivo with recombinant IL 2. AB - Purified recombinant human interleukin 2 (RIL 2) derived from E. coli containing the inserted gene encoding for IL 2 was administered to 20 patients with a variety of malignancies. Toxicity was dose related and included fever, chills, malaise, arthralgias, myalgias, and unexpectedly, weight gain related to marked fluid retention. All patients receiving more than 10(5) U/kg total cumulative dose developed evidence of fluid retention, and all patients requiring discontinuance of RIL 2 (11/20) received total doses of between 2.54 X 10(5) U/kg to 15.4 X 10(5) U/kg. The limiting dose with this preparation was 3000 U/kg/hr by continuous administration or 10(6) U/kg by bolus administration. IL 2 was rapidly cleared from the plasma, with a half life of 6.9 min, and a later delayed clearance was consistent with a two-compartment model, with slower release from the extravascular space back into the plasma compartment. A marked change in lymphoid cells in the periphery was noted with an early depletion of all lymphoid cells, followed by an expansion of such cells with continuous IL 2 administration. A twofold to 16-fold expansion of total lymphoid cells in the peripheral blood could be demonstrated. TAC+ cells representing up to 25% of the circulating peripheral blood mononuclear cells could be demonstrated with 3 wk of continuous RIL 2 administration. Interferon-gamma levels increased in patients treated with IL 2. Precursors of lymphokine-activated killer cells generated under standard conditions were depleted within 2 to 3 min after IL 2 administration, but repopulated the peripheral blood after 7 to 10 days of continuous IL 2 administration. No tumor regression was seen in any of the cancer patients treated with IL 2 alone. PMID- 2993420 TI - The role of interleukin 1 (IL 1) in tumor-NK cell interactions: correction of defective NK cell activity in cancer patients by treating target cells with IL 1. AB - This study examined the importance of interleukin 1 (IL 1) in the large granular lymphocyte (LGL)-target cell interaction. K562 target cells when treated with highly purified human IL 1 for 1 hr bound greater numbers of LGL than untreated cells. LGL from patients with hepatocellular carcinoma (HCC) that bound few untreated K562 cells, attached to considerably increased numbers of IL 1-treated target cells. Cytotoxicity of LGL against target cells could similarly be increased by pulsing the latter cells with IL 1, and defective cytotoxicity of LGL from HCC patients could be corrected by treating the target K562 cells with IL 1. Lysis of PLC/PRF/5 cells, Yac-1 cells, and normal skin fibroblasts could also be increased by treatment with IL 1 for 1 hr. The enhanced binding and cytotoxicity of IL 1-treated target cells was only observed when the latter cells were preincubated with IL 1 at 37 degrees C, and was not evident at 4 degrees C. Furthermore, the IL 1-mediated effect could be abolished by treating the target cells with cycloheximide before the IL 1 pulse, or by adding rabbit anti-human IL 1 together with the IL 1. These results indicate that IL 1 affects a variety of target cells and increases their ability to bind and be lysed by enriched LGL. They demonstrate, furthermore, that defective natural cytotoxicity by the LGL of patients with advanced malignant disease can be corrected in vitro by treating the target cells with IL 1. PMID- 2993419 TI - In vitro maturation of B cells in chronic lymphocytic leukemia. I. Synergistic action of phorbol ester and interleukin 2 in the induction of Tac antigen expression and interleukin 2 responsiveness in leukemic B cells. AB - Recent evidence indicates that interleukin 2 (IL 2), formerly thought to serve as growth factor exclusively for activated T cells, is directly involved in human B cell differentiation. We have investigated the role of IL 2 and IL 2 receptors (as defined by monoclonal anti-Tac antibody) in the phorbol ester-induced in vitro maturation of leukemic B cells from patients with chronic lymphocytic leukemia (CLL). Peripheral blood lymphocytes from B cells from CLL patients with high (greater than 10(5)/microliters) white blood cell counts were depleted of residual T lymphocytes and low-density cells (primarily macrophages) by consecutive steps of E rosetting, complement-mediated lysis of OKT3+ and OKT4+ cells, and Percoll density gradient centrifugation. No OKT3+ T cells were detectable in these cell populations before or after culture. When incubated for 3 days with phorbol ester plus recombinant human IL 2 (rIL 2), 12 to 57% of highly purified B cells from four of five tested patients expressed Tac antigen. Both phorbol ester and rIL 2 were required for maximal Tac antigen expression. Functional studies revealed that phorbol ester-activated (but not resting) CLL B cells responded to rIL 2 with [3H]thymidine incorporation and with enhanced secretion of IgM. Tac+ B cells were isolated in two cases on a fluorescence activated cell sorter. In one patient, stimulation of Tac+ B cells with rIL 2 resulted in enhanced [3H]thymidine incorporation but no change in IgM secretion, as compared with Tac- B cells; in the second patient, stimulation of Tac+ B cells with rIL 2 did not result in [3H]thymidine uptake, but did result in significant IgM secretion. These findings indicate that certain leukemic B lymphocytes can be induced to express IL 2 receptors and respond to IL 2. The use of resting clonal B cell populations arrested at distinct stages of differentiation may help to better define the stage(s) at which IL 2 acts directly on B cells to induce proliferation and/or terminal differentiation. PMID- 2993421 TI - Alteration in antibody reactivity with foot-and-mouth disease virus (FMDV) 146S antigen before and after binding to a solid phase or complexing with specific antibody. AB - This paper describes the reactions of a number of monoclonal antibodies produced against purified whole virions of foot-and-mouth disease virus in 3 different enzyme immunoassay systems. The first system used whole virus bound non covalently to microplates; the second used whole virus trapped by a polyclonal antibody which was bound to microplates; and the third allowed the monoclonal antibodies to react with the whole virions in suspension (liquid phase) before trapping by the solid-phase-bound polyclonal antibody. Different reactions with panels of monoclonal antibodies were observed depending on which system was used. Such variations in reactivity give an insight into the alterations in the expression of virus epitopes in the different enzyme immunoassay systems. The reactions of selected monoclonal antibodies were used to illustrate these changes and the results compared to those obtained in similar systems using polyclonal antisera produced against isolated virion polypeptides. PMID- 2993422 TI - Theophylline induced non specific suppressor activity in human peripheral blood lymphocytes. AB - It is wellknown that theophylline yields phenotypic changes on suppressor cells. In the present study we investigated the possibility that theophylline could directly induce a suppressor activity on a lymphocyte subpopulation. We observed that a short preincubation (120 min at 37 degrees C) with theophylline (1mM) activates human peripheral blood lymphocytes to suppress mitogenic response of autologous cells. This activity was not evident on a T cell subpopulation depleted of theophylline-sensitive (T-sens) lymphocytes. Theophylline mediated suppressor activity is only present in the Concanavalin A stimulated cultures, thus suggesting a synergism between Concanavalin A and theophylline in the expression of non specific suppression. Moreover we observed that after a 24 hrs preincubation of lymphocytes in complete culture medium there was a complete loss of theophylline-induced suppression. Such a preincubation time also produced a decrease in the theophylline-mediated enhancement of intracellular 3', 5' cyclic adenosine monophosphate levels and the impairment of E-rosette formation, suggesting that theophylline acts mainly on a "short-lived" suppressor lymphocyte subset. PMID- 2993423 TI - The postviral fatigue syndrome--an analysis of the findings in 50 cases. AB - The clinical, pathological, electrophysiological, immunological and virological abnormalities in 50 patients with the postviral fatigue syndrome are recorded. These findings confirm the organic nature of the disease. A metabolic disorder, caused by persistent virus infection and associated with defective immunoregulation, is suggested as the pathogenetic mechanism. PMID- 2993424 TI - Acute Epstein-Barr virus infection and diffuse large-cell lymphoma. AB - The antibody response to Epstein-Barr virus (EBV) has not been fully investigated in previous cases of diffuse large-cell lymphoma associated with infectious mononucleosis. It is not clear, therefore, whether acute EBV infection is related to this malignant condition. In this paper, a case of diffuse large-cell lymphoma accompanied by acute EBV infection is described. PMID- 2993425 TI - Survey of the prevalence of AIDS-associated virus (LAV) infection in Japan. AB - An indirect immunofluorescence method was developed for the assay of antibodies to lymphadenopathy-associated virus (LAV) which is known to be associated with the aetiology of the acquired immunodeficiency syndrome (AIDS). Samples of serum or plasma from 1353 healthy volunteer blood donors, 53 homosexual males, 29 patients who had received multiple blood transfusions and 163 haemophiliacs in Japan were tested for antibody to LAV. Results showed that 47 (29%) of the haemophiliacs, who had been treated largely with factor VIII or IX produced in the USA, were anti-LAV antibody positive, whereas all other subjects were anti LAV antibody negative. The incidence of antibodies to Adult T-cell leukaemia virus or Human T-cell leukaemia virus I (ATLV or HTLV-I) in these subjects was high. PMID- 2993427 TI - Psoralens increase the concentration of cyclic AMP in human cells in vitro. AB - 8-Methoxypsoralen (8-MOP) and 4,5',8-trimethylpsoralen (TMP) were assessed for their effects on cyclic AMP (cAMP) accumulation in normal human fibroblasts, epithelial cells, and mononuclear leukocytes in vitro. In the absence of ultraviolet A radiation, 8-MOP and TMP caused an increase in the concentration of cAMP, but not cyclic GMP, in all of these cells. The effect was seen within 1 min and was sustained over control levels for at least 6 h. At 15 min, the highest dose of 8-MOP or TMP increased the cAMP concentration of fibroblasts by 52% and 69%, respectively. In mononuclear leukocytes, 8-MOP led to a 142% increase in cAMP after 20 min. Several analogs of 8-MOP and TMP had no effect on the cyclic nucleotide system of the cells. The pattern of the time-dependent increase in cAMP indicated that the effect was probably due to an inhibition of phosphodiesterase, rather than an activation of adenylyl cyclase. These results demonstrate that clinically effective psoralens, in the absence of UV radiation, can alter the cAMP concentration of human fibroblasts, epithelial cells, and mononuclear leukocytes in vitro. PMID- 2993426 TI - Challenges in clinical virology (1958-1974): a personal viewpoint. AB - Some of the main challenges I encountered in clinical virology from 1958-1974 are described. In the earlier years there were two main areas of exploration, namely, neurological and respiratory infections. Later, tests for hepatitis and for rubella were developed and these diseases became of increasing importance. PMID- 2993428 TI - Cytoplasmic vitamin A binding proteins in chick embryo dermis and epidermis. AB - Excess vitamin A has striking morphologic and developmental effects on chick embryo skin. While cytoplasmic retinoic acid-binding protein (CRABP) was known to be abundant in chick embryo skin, neither quantitative values nor the distribution between dermis and epidermis have been established. We determined CRABP levels in collagenase-separated dermis and epidermis from 8-day-old embryos using specific binding of all-trans-[11-3H]retinoic acid in cytosols prepared from gram quantities of these tissues. The level of CRABP in dermis was twice the level in epidermis whether calculated on the basis of wet weight, cytosol protein, or DNA. When averaged over many preparations, 3 times as much dermis as epidermis was recovered from a single piece of skin. Therefore, the dermis contained 85% of the extremely high CRABP levels found in collagenase-treated skin, while epidermis contributed only 15%. Cytoplasmic retinol binding protein (CRBP) was also detected in chick embryo skin, but the binding was low and the levels in epidermis and dermis were not significantly different. The amount of CRABP in chick embryo skin (1600 pmol/g wet weight or 100 pmol/mg cytosol protein) is the highest level reported in any tissue and suggests an important role for vitamin A in the normal development and maturation of skin. PMID- 2993429 TI - Epidemiology and transmission of cytomegalovirus. PMID- 2993430 TI - Epidemiology of cytomegaloviral infection in a heterogeneous population of pregnant women. AB - Cervical cultures for cytomegalovirus (CMV) and samples of blood for antibody to CMV were obtained from 1,129 pregnant women: 57% of the women had antibody to CMV, and 14% of seropositive women shed virus. Logistic regression analysis showed that seropositivity correlated with lower socioeconomic status, birth outside North America, multigravidity, older age, history of abnormal cervical cytology, infection with Trichomonas vaginalis, a first pregnancy at less than or equal to 15 years of age, and greater total numbers of sex partners. Thus, past exposure to CMV relates both to sociocultural factors and to sexual behavior. Absence of such risk factors identifies women who are at highest risk for primary infection with CMV during pregnancy. Culture positivity in seropositive women was independently associated with younger age, later stages of pregnancy, and race. Among seropositive women less than or equal to 21 years of age, 35% shed CMV in the third trimester, a finding of epidemiological importance with regard to perinatal transmission. PMID- 2993431 TI - Experimental parvoviral infection in humans. AB - Healthy adult volunteers were inoculated intranasally with human parvovirus obtained from an asymptomatic blood donor. One week after inoculation, intense viremia was observed in seronegative volunteers, accompanied by a mild illness with pyrexia, malaise, myalgia, itching, and excretion of virus from the respiratory tract. In the following week hematologic studies revealed reticulocytopenia with an associated slight drop in hemoglobin concentration, lymphopenia, neutropenia, and a drop in platelet counts. At 17-18 days after inoculation a second-phase illness with rash and arthralgia lasting three to four days occurred in three of four infected volunteers. This study confirms the etiologic role of human parvovirus in erythematous rash illness, with the second phase illness being consistent with adult cases of erythema infectiosum. Moreover, the hematologic changes associated with infection support the hypothesis that the same virus is responsible for the temporary arrest of erythropoiesis that leads to aplastic crisis in persons with chronic hemolytic anemia. PMID- 2993432 TI - Humoral immunity in infants with gastroenteritis caused by human calicivirus. AB - To assess the serum immune response in infants with gastroenteritis caused by human calicivirus (HCV), we tested a total of 165 serum specimens for antibodies to HCV by a radioimmunoassay blocking (RIA-BL) test. The specificity of the RIA BL test for antibody to HCV was established as follows: paired sera from patients with HCV gastroenteritis revealed significant rises in antibody to HCV, whereas there was no significant change in RIA-BL titer to HCV in paired sera collected from the same individuals when infected with other gastroenteritis viruses. We analyzed 41 paired sera from patients and from healthy contacts involved in an outbreak of HCV gastroenteritis. All 23 infants without preexisting antibody revealed a sharp rise in HCV titer, and 18 (73%) of 23 of them developed illness. Fifteen of 18 infants with preexisting antibody also revealed a significant rise in HCV antibody, but only 3 (16.7%) of 18 became ill (P less than .01). These data suggest that the presence of serum antibody may correlate with resistance to illness in HCV gastroenteritis. PMID- 2993433 TI - Human monoclonal antibodies neutralizing varicella-zoster virus. AB - Hybridomas secreting human monoclonal antibodies to varicella-zoster virus were produced by fusing B cells of a patient recovering from acute varicella infection with a human-mouse cell line. Two hybrid lines have continued to secrete IgG1, one with kappa and the other with lambda chains, for at least 12 months. Each antibody neutralizes virus infectivity between 1-5 micrograms of partially purified immunoglobulin/ml, each shows a different pattern of immunofluorescent staining of virus-infected cells, and one identifies three viral proteins with molecular weights of 60,000, 95,000, and 97,000. PMID- 2993434 TI - Detection of rotaviruses by nucleic acid hybridization with cloned DNA of simian rotavirus SA11 genes. AB - We developed a dot-blot hybridization assay to detect rotaviral RNA sequences in tissue culture or in clinical samples. 32P-labeled cloned cDNA probes of the simian rotavirus SA11 specifically detected rotaviral RNA sequences and were more sensitive for detecting SA11 than was the commercial enzyme-linked immunosorbent assay Rotazyme test. A full-length probe of SA11 gene 6 detected 2.5 X 10(5) SA11 particles or approximately 0.27 ng of purified SA11 dsRNA. Combined probes from genes 6 and 9 detected 0.135 ng of purified SA11 dsRNA. The assay detected group A rotaviruses from different subgroups and serotypes, but the sensitivity of RNA detection varied from 0.5 to 31 ng when RNA from heterologous strains of virus was analyzed. An analysis of coded stool samples correctly identified 31 (91%) of 34 samples positive for rotavirus by electron microscopy and 100% of 26 samples negative for rotavirus by electron microscopy. Preliminary experiments also showed the assay has potential to directly characterize (subgroup and serotype) rotaviral isolates. PMID- 2993435 TI - Comparative susceptibility of group B streptococci and Staphylococcus aureus to killing by oxygen metabolites. AB - Group B streptococci (GBS) lack catalase, and they produce and release H2O2;thus, they should be readily killed by phagocytes with a diminished respiratory burst. Surprisingly, although strains of Staphylococcus aureus were killed at H2O2 concentrations greater than 0.5 mM, GBS strains were killed only at concentrations greater than 5mM. In contrast, GBS were killed by hydroxyl radicals generated by the xanthine oxidase-acetaldehyde system at O2 fluxes greater than or equal to 3.5 nmol/ml per min, whereas O2 fluxes greater than or equal to 10 nmol/ml per min were required to kill the S. aureus strains. Results with virulent and laboratory strains of GBS were similar. The differences in susceptibility of GBS and S. aureus seemed to correlate with differences in content of endogenous oxygen-metabolite scavengers. GBS contained approximately 100-fold more glutathione and approximately 20-fold more glutathione reductase than did S. aureus, whereas S. aureus was rich in catalase that GBS lacked. GBS that were grown in buthionine sulfoximine, however, contained 87% less glutathione than did controls but were not more susceptible to killing by H2O2 or the xanthine oxidase-acetaldehyde system. Similarly, the relative susceptibility of GBS to tert-butyl hydroperoxide and H2O2 paralleled that of S. aureus. Thus, inherent differences in susceptibility of vital cellular functions to oxidative damage rather than content of oxygen-metabolite scavengers may account for the differences in susceptibility of GBS and S. aureus. PMID- 2993436 TI - Cytomegalovirus vaccine virus (Towne strain) does not induce latency. PMID- 2993437 TI - Anamnestic response in fecal IgA antibody production after rotaviral infection of infants. PMID- 2993438 TI - Disinfection and inactivation of the human T lymphotropic virus type III/Lymphadenopathy-associated virus. PMID- 2993439 TI - Herpes simplex virus immunology: problems, progress, and promises. PMID- 2993440 TI - Viral inhibition of lymphocyte proliferative responsiveness in patients suffering from recurrent lesions caused by herpes simplex virus. AB - Herpes simplex viruses (HSV) are able to prevent the lectin-driven mitogenesis of coincubated human peripheral blood lymphocytes. This abrogation of responsiveness is independent of infection and can be obtained by using ultraviolet-inactivated as well as live virus particles. Lymphocytes from patients prone to frequent recurrences of HSV-induced lesions were more susceptible to inhibition of proliferative responsiveness than were cells from either individuals seronegative for HSV type 1 and HSV type 2 or individuals subject only to infrequent recurrences of HSV-induced disease. Lymphocytes from all patients tested were more susceptible to viral abrogation of mitogen responsiveness during preepisodic and acute as opposed to convalescent and postepisodic periods. When exogenous T cell growth factor was added to lymphoid cell cultures that had been coincubated with HSV, the cells were generally able to overcome the inhibition of responsiveness that would otherwise occur. However, this restoration of responsiveness occurred less efficiently in cells from patients with acute HSV induced lesions and in cells from patients with frequent recurrences of lesions. PMID- 2993442 TI - The epidemiology of cytomegaloviral infection in women attending a sexually transmitted disease clinic. AB - To test the hypothesis that cytomegalovirus (CMV) is sexually transmitted, we examined the association of CMV infection with indices of sexual activity in 347 women attending a sexually transmitted disease (STD) clinic. Stepwise multivariate logistic regression analysis showed that seropositivity to CMV (complement-fixation antibody titer, greater than or equal to 1:8) was most closely associated with number of sex partners in the subjects' lifetime (P less than .0001), young age at first sexual intercourse (P = .0002), and nonwhite race (P = .0007). Among seropositive women, cervical shedding of CMV was most strongly associated with younger age (P = .0001) and the presence of cervical chlamydial infection (P = .016). Among 84 seronegative women followed up for a mean of 18.4 weeks, 11 (13%) developed primary CMV infections, an annual incidence of 37%. Sexual contact seems to be an important mode of acquisition of CMV in some young women. PMID- 2993441 TI - Interferon-induced 2'-5' oligoadenylate synthetase during interferon-alpha therapy in homosexual men with Kaposi's sarcoma: marked deficiency in biochemical response to interferon in patients with acquired immunodeficiency syndrome. AB - Titers of circulating interferon (IFN) and the activity of 2'-5' oligoadenylate (2-5A) synthetase, an enzyme specifically induced by IFN, were measured in 28 homosexual men with acquired immunodeficiency syndrome (AIDS) who received one- to six-month courses of antineoplastic therapy with IFN-alpha and in homosexual and heterosexual controls. Fifteen of the patients and two of seven healthy homosexual men had high endogenous levels of 2-5A synthetase. IFN therapy induced further increases in this enzyme in only 10 of the 28 patients with AIDS. Furthermore, peripheral blood cells from all but one of the patients with AIDS and homosexual controls tested were markedly deficient in their ability to respond to IFN in vitro, as measured by increased levels of 2-5A synthetase. We did not find a statistical correlation between cytomegalovirus viremia and pretherapy endogenous circulating IFN, nor any apparent correlation between disseminated infection with cytomegalovirus and either basal levels of 2-5A synthetase or changes in enzyme level during therapy. Pretherapy circulating IFN was significantly correlated with progressive Kaposi's sarcoma during therapy, but rises in levels of 2-5A synthetase were not sufficient to predict a good clinical response. PMID- 2993443 TI - Congenital defects due to reactivation of latent murine cytomegaloviral infection during pregnancy. PMID- 2993444 TI - Activation of latent murine cytomegalovirus in cardiac explant and cell cultures. PMID- 2993445 TI - Deficient, HLA-restricted, cytomegalovirus-specific cytotoxic T cells and natural killer cells in patients with the acquired immunodeficiency syndrome. PMID- 2993446 TI - Rapid detection of human cytomegalovirus in the urine of humans. PMID- 2993447 TI - The S1 gene from reovirus type 1 is required for immunosuppression. PMID- 2993448 TI - Rotavirus in children. PMID- 2993449 TI - Comparison of the rotazyme assay with an avidin-biotin-amplified dot immunobinding assay for detecting rotaviruses. PMID- 2993450 TI - Serological heterogeneity of CMV isolates with a monoclonal antibody. PMID- 2993451 TI - Central European tick-borne encephalitis: assessment of risk for persons in the armed services and vacationers. PMID- 2993452 TI - [Exfoliative cytology of mixed mesodermal tumors of the uterus--review of cytologic smears sampled prior to therapy]. AB - Nine cases of mixed mesodermal tumor of the uterus were reviewed cytologically. Abnormal cytologic findings were seen in all cases, but malignant cells were detected in only 17 of 30 cytologic smears sampled prior to the therapy (2 of 7 pool smears, 2 of 3 portio rubbing-smears, 6 of 8 endocervical smears and 7 of 12 tumor-rubbing smears). The most commonly encountered cellular components were epithelial malignant cells which showed a close resemblance to those derived from uterine carcinomas, but tended to have multiple nuclei. In other cytologic components, a rhabdomyosarcoma was characterized by large, isolated, round or strap-shaped malignant cells with granular cytoplasms, bizarre and eccentric nuclei and one or more prominent nucleoli. Another infrequent cellular component was a fibrogenic sarcomatous cell which had an illdefined lacy cytoplasm and single or multiple nuclei containing rather finely granular chromatin, and occasionally formed a cluster. In several cytologic specimens, there were also seen multinucleated giant cells which were not in keeping with their histologic origins. PMID- 2993453 TI - Human papillomavirus DNA in female condylomata. AB - We have studied the presence and state of human papillomavirus (HPV) genomes in condylomata acuminata of the vulva, vagina, and cervix and in flat condylomas of the cervix from Japanese females by DNA hybridization with HPV 6 and 11 DNAs. Unintegrated HPV 6-related DNA (HPV 6 or 11 DNA) was detected in 55 out of 60 condylomata acuminata and in 1 out of 4 flat condylomas under stringent hybridization conditions. The HPV DNAs were identified as HPV 6 DNA in 29 condylomata acuminata (20 vulvar, 3 vaginal, and 6 cervical samples) and 1 flat condyloma and as HPV 11 DNA in 6 condylomata acuminata (6 vulvar samples). The data indicate close association of HPV types 6 and 11 with condylomata. PMID- 2993455 TI - Synovial sarcoma: with radial nerve involvement. AB - An unusual case of radial nerve involvement at the elbow by a synovial sarcoma is reported. The patient was a 16-year-old Caucasian girl presenting with signs of a radial nerve palsy. Treatment was by surgical excision of the tumour and grafting the defect in the radial nerve. The histological diagnosis was a synovial sarcoma. Recovery of function was almost complete eight months later. PMID- 2993454 TI - [Studies on the medical treatment of non-operable insulinoma. Clinical usefulness of diphenylhydantoin-calcium antagonist]. PMID- 2993456 TI - Carpal coalition and Sprengel's shoulder in Poland's syndrome. PMID- 2993458 TI - Effects of dietary calcium restriction on 1,25-dihydroxyvitamin D3 net synthesis by rat proximal tubules. AB - We measured in vitro 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) production by kidney proximal tubules prepared by Percoll density centrifugation from male and female rats. 1,25(OH)2D3 in tubule extracts was determined by a sensitive and specific radioreceptor assay. Ingestion of diets adequate in vitamin D3 and containing either normal calcium (1.2% Ca, NC), reduced calcium (0.6% Ca, RCD) or low calcium (0.002% Ca, LCD) increased 1,25(OH)2D3 net synthesis (for male rats, NC vs. RCD vs. LCD 1.8 +/- 0.1 SEM vs. 9 +/- 2 vs. 17 +/- 2 pmol/mg protein/20 min; P less than 0.05 for all comparisons). At either level of reduced calcium intake, tubules from male rats produced more 1,25(OH)2D3 than tubules from females. Serum 1,25(OH)2D3 and tubule cyclic adenosine monophosphate (cAMP) content rose in parallel with progressive dietary calcium restriction, and males had higher circulating 1,25(OH)2D3 and tubule cAMP content than females at each level of reduced calcium intake. L-Epinephrine (10(-4) mol/L), in vitro, increased tubule accumulation of 1,25(OH)2D3 and cAMP. Yohimbine, and alpha 2-receptor antagonist, blocked this response, whereas prazosin was without effect. Increased 1,25(OH)2D3 net synthesis by tubules from male vs. female rats partly explains the higher serum levels and enhanced mineral conservation demonstrated previously in male rats. Preparation of proximal tubules from vitamin D-replete rats permits studies in vitro of 1,25(OH)2D3 production and regulation under more physiologic conditions in which parathyroid hormone, inorganic phosphorus, and calcium may be varied independently. PMID- 2993459 TI - Preparation of pure hepatocytes and reticuloendothelial cells in high yield from a single rat liver by means of Percoll centrifugation and selective adherence. AB - A rapid method for mass isolation of functionally intact hepatocytes and reticuloendothelial cells from a single rat liver is described. The technique is based on collagenase perfusion of the liver, isopycnic sedimentation in Percoll, and selective adherence of the cells. The Kupffer cells (KC) attach and spread on glass or plastic in serum-free medium 15 min following seeding. Cultures of KC are 90%-95% pure with about 5% liver endothelial cells (LEC), less than 1% parenchymal cells (PC) and a maximum of 5% stellate cells (SC). The LEC adhere and spread on fibronectin 60-120 min following seeding, forming cultures that are contaminated with 5-10% SC and less than 1% KC and PC. The yield of plated LEC is 50-60 X 10(6) per 200-g rat. Ultrastructural analysis shows that Percoll does not associate with the cells during the separation procedure. PMID- 2993457 TI - Hyperoxic lung injury: biochemical, cellular, and morphologic characterization in the mouse. AB - Controversy exists over the mechanism by which oxygen produces lung damage. This study was designed to characterize the biochemical, cellular, and morphologic responses of mice exposed to 100% oxygen and thereby provide the basis for investigating the mechanisms producing the lung damage as well as for evaluating treatment strategies. After 1 day of exposure, plasma lactate dehydrogenase (LDH) activity and LDH isoenzyme 1, 2, and 3 concentration increased, whereas the level of isoenzyme 1 recovered from the lung, heart, and kidney decreased. On day 2, the level of these plasma isoenzymes increased further, whereas isoenzyme 1 concentration remained decreased only in the lung. In addition, the number of cells obtained by bronchoalveolar lavage (BAL) doubled, and electron microscopy revealed type 1 cell and endothelial cell damage. On day 3, the amount of BAL protein doubled, BAL angiotensin-converting enzyme (ACE) activity increased sevenfold, plasma ACE activity decreased 39%, and electron microscopy revealed extensive cell damage. On day 4, the injury was much worse and was associated with a 10-fold increase in the number of BAL cells, nearly all of which were polymorphonuclear leukocytes (PMNs). On day 5, greater than 65% of the mice were dead. The data suggest that in mice (1) an increase in plasma LDH activity and a shift in its isoenzyme pattern are sensitive markers of hyperoxic lung damage, (2) the initial injury produced by oxygen is independent of PMNs, and (3) the damage to alveolar epithelial cells and endothelial cells is severe and occurs at the same time. Studies using this well-characterized model can now be designed to further define the mechanisms that initiate hyperoxic lung damage and that contribute to its progression. PMID- 2993460 TI - Phagocytosis by receptors for C3b (CR1), iC3b (CR3), and IgG (Fc) on human peritoneal macrophages. AB - Human peritoneal macrophages (HPM) obtained via laparoscopy were examined for the presence and functional capacity of complement and Fc receptors. Between 5 and 20 ml of peritoneal fluid containing 1-2 X 10(6) macrophages/ml was available for each study. Macrophages made up 80-95% of the cells in the fluid. Fc and C3 receptors on HPM were characterized by rosette formation with, and phagocytosis of, IgG- and C3-coated sheep erythrocytes (E). ElgG were bound by 82% and ingested by 63% of HPM, with 4-15 E ingested/HPM. The HPM formed rosettes with EC3b (56%) and EC3bi (71%) but not EC3d,g or EC3d. Antibodies to complement receptors type 1 (CR1) and type 3 (CR3) inhibited rosette formation with EC3b and EC3bi, respectively, indicating that HPM possessed separate and distinct receptors for the C3b and iC3b ligands. In 60% of the samples studied, HPM demonstrated the ability to ingest both EC3b and EC3bi, as well as ElgG. Because of the heterogeneous nature of the cells obtained in peritoneal fluid, due to their progressive change from monocytelike cells into mature macrophages, HPM were separated by 1 g velocity sedimentation into fractions of increasing maturity. They were then examined for phagocytosis via Fc and complement receptors. Fc receptor mediated phagocytosis occurred throughout the monocyte-to macrophage maturation sequence, while the ability of HPM to ingest via CR1 and CR3 was maturation dependent, with ingestion via CR3 occurring before CR1, in a manner analogous to in vitro differentiation of monocyte-derived macrophages. PMID- 2993461 TI - Isolation of canine neutrophil plasma membranes. AB - This paper describes the isolation of plasma membrane vesicles formed by nitrogen cavitation of canine neutrophils. Plasma membranes from disrupted cells were separated from other membranes and organelles by Percoll-density gradient centrifugation. Transmission electron microscopic examination of membrane preparations chromatographed on either Sephacryl S-1000 or Sepharose 4B revealed that two populations of plasma membrane vesicles were formed: large (176 +/- 22nm), and small (119 +/- 11nm). Purified large vesicles were separated from Percoll and contaminating cytosol by Sephacryl S-1000 chromatography. Small vesicles were obtained free of Percoll by recavitating purified large vesicles. Problems encountered due to the presence of a soluble 5'-nucleotidase inhibitor also are discussed. Large and small membrane vesicles were separated into adherent and non-adherent populations by affinity chromatography on either concanavalin A-Sepharose or lentil lectin-Sepharose columns. PMID- 2993462 TI - The chemotactic response of granulocytes to the low molecular weight chemoattractants f-MLP, C5f, and LTB4 is dependent on chemokinetic factors. AB - The dependence of the low molecular weight chemoattractants (LMCs) formylmethionyl-leucylphenylalanine (f-MLP), C5f, and leukotriene B4 (LTB4) on albumin to express their chemotactic activity towards granulocytes (PMNs) was investigated in order to study the required qualities of albumin and if albumin could be replaced by any other proteins. The results demonstrated that the supporting effect of isolated albumin was dependent on the method of purification. Only isolated albumin exposed to ethanol precipitation during the purification procedure supported the chemotactic effect of LMCs. The albumin preparation that supported the effect of LMCs also mediated a chemokinetic effect on PMN migration. Albumin isolated by methods other than ethanol precipitation neither exerted a chemokinetic effect nor supported the chemotactic effect of LMCs. Heated, normal serum and isolated alpha 1-antitrypsin supported the chemotactic activity of LMCs and also mediated a chemokinetic effect on PMN migration. The present investigation suggests an important role for the chemokinetic factors, since it is indicated that their presence is necessary for the chemotactic response of PMNs to the low molecular weight chemoattractants C5f; LTB4, and f-MLP. PMID- 2993463 TI - Specific binding of high density lipoprotein (HDL3) is not related to sterol synthesis in rat intestinal mucosa. AB - There is good evidence that high density lipoprotein (HDL) is involved in the flux of cholesterol into the cells of some organs and out of the cells of other tissues. Because we have previously found that HDL is bound specifically by mucosal cells of the small intestine, we have examined the possibility that this was associated with regulation of cholesterol flux. We have, therefore, compared the specific binding of 125I-labeled HDL3 with cholesterol synthesis in mucosal cells obtained from rats that had been treated to alter intestinal cholesterol metabolism. The rate of sterol synthesis measured in tissue slices, by the incorporation of [3H]water into sterols, was altered up to fivefold by treatment with cholestyramine (to induce bile salt loss), by surformer treatment (to reduce absorption of cholesterol), and by biliary diversion. Yet the capacity of mucosal cells to bind, internalize, and degrade 125I-labeled HDL3 was unchanged. Cholesterol feeding influenced neither the interaction of 125I-labeled HDL3 with cells nor the rate of sterol synthesis. Furthermore, the interactions of 125I labeled HDL3 with mucosal cells isolated from the proximal and distal halves of the intestine or between the upper and lower villus cells were similar, despite differences in sterol synthesis. These data suggest that, in rat intestine, the specific binding of HDL is not related to sterol synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993464 TI - Modification of human high density lipoprotein (HDL3) with tetranitromethane and the effect on its binding to isolated rat liver plasma membranes. AB - Apolipoprotein E-free high density lipoproteins (HDL) bind to various cells and cell membrane preparations, with properties typical of ligand-receptor interactions. In order to further characterize the binding sites and to investigate the functional role of binding, a chemically modified HDL without the specific binding properties would be highly desirable. We have reacted human HDL3 with tetranitromethane, a relatively specific nitrating reagent for tyrosine residues, in 50 mM Tris HCL buffer, pH 8.0, and at a reagent concentration 10 times the molar excess of tyrosine residues. The resulting nitrated HDL3 completely lost its ability to bind to high affinity saturable binding sites of rat liver plasma membranes, as determined by competitive binding with 125I labeled HDL3, and also by direct binding assays using 125I-labeled nitrated HDL3. Although nitrated HDL3 did not bind to the high affinity saturable binding sites, it bound to the membranes, but the binding was not saturable, and was not competed for by unlabeled nitrated HDL3. On agarose gel electrophoresis, pH 8.6, the nitrated HDL3 moved ahead of the control HDL3, indicating an increase in negative charges in the molecule. No difference in size was noted in the nitrated HDL3 when analyzed either by negative stain electron microscopy or by gel filtration chromatography. Spectroscopic analysis of the nitrated HDL3 at pH 8.0 revealed a prominent absorption with maximum at around 360 nm, but none in the region expected for nitrotyrosine residues. At pH 10.0, however, the nitrated HDL3 showed an absorption band with a maximum at around 440 nm, possibly related to nitrotyrosine residues. Nitrotyrosine was detected in the nitrated HDL3 on amino acid analysis. Comparison of the amino acid analysis of the nitrated HDL3 and control HDL3 showed no difference in composition of any of the amino acids except tyrosine; tyrosine content was reduced more than 90% in the nitrated HDL3. SDS-polyacrylamide gel electrophoresis analysis of apoproteins of nitrated HDL3 revealed changes in apolipoprotein profile. Bands corresponding to the apolipoproteins of the starting HDL3 almost disappeared and a series of new bands appeared at the high molecular weight region of the gel, indicating extensive cross-linking of apolipoproteins during the reaction. In addition, a substantial amount of phospholipids and cholesteryl esters, but not unesterified cholesterol, was found covalently linked, possibly through the unsaturated centers of the fatty acid chains, to apolipoproteins.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993465 TI - [Microcirculation, mesotherapy and thermography]. PMID- 2993466 TI - Thrombotic thrombocytopenic purpura associated with small cell lung carcinoma. PMID- 2993467 TI - Opisthorchiasis and its associated diseases. PMID- 2993468 TI - Influence of cyclic AMP and hydrolysis products on cell type regulation in Dictyostelium discoideum. AB - We describe the effect of cyclic AMP on regulation of the proportion of prespore and prestalk cells in Dictyostelium discoideum. Prespore and prestalk cells from slugs were enriched on Percoll density gradients and allowed to regulate in suspension culture under 100% oxygen. The transition of prespore to prestalk cells is blocked by cAMP, while cAMP phosphodiesterase and caffeine cause a decrease in the number of prespore cells. This suggests that extracellular cAMP plays a role in cell type proportioning by inhibiting the conversion of prespore to prestalk cells. Low concentrations of cAMP prevent the conversion of prestalk to prespore cells; the same effect is seen with hydrolysis products of cAMP, 5 AMP, adenosine and also adenine. We suggest that, when low concentrations of cAMP are added to regulating cells, the cAMP itself is quickly broken down and the breakdown products thereafter inhibit the prestalk-to-prespore conversion. The relevance of these findings is discussed in the context of an non-positional double-negative feedback model for cell type homeostasis. PMID- 2993469 TI - Restriction fragment length polymorphism of the gene encoding the alpha chain of the human T cell receptor. AB - Two allelic forms of the T cell antigen receptor alpha chain gene were discerned by restriction fragment length polymorphism (RFLP) employing the T cell antigen receptor alpha chain probe pGA5, and the restriction enzyme Bgl II. Analysis revealed that the polymorphic fragments are detected by a probe specific for the constant region exon of the T cell antigen receptor alpha chain gene. Furthermore, the polymorphic fragments were shown to segregate within families. The two allelic forms yield two homozygous states, 3.2/3.2 and 2.9/2.9, at a frequency of 76.5 and 2.9%, respectively, within the normal population. The heterozygous state was observed in 20.6% of the population. The discovery of allelic forms of both the alpha and beta chains of the T cell antigen receptor genes may provide a unique opportunity to study heritable markers of T cell function in several human diseases. PMID- 2993470 TI - Growth factor-mediated tumor cell proliferation in hairy cell leukemia. AB - Leukemic B cells from seven patients with hairy cell leukemia (HCL), six of which contained the Tac antigen, were assayed in vitro for growth factor-mediated cell proliferation. The HCL cells showed typical phenotypic profiles by monoclonal antibody analysis. The tumor cells, which do not grow spontaneously in vitro, were found to proliferate in all but one case in response to partially purified B cell growth factor (BCGF) without anti-mu or Sac activation. Recombinant interleukin 2 however produced only a marginal response and could not support leukemic cell growth in vitro. BCGF, however, did stimulate in vitro cell growth and supported the establishment of continuous (greater than 60 d in vitro) in four of the seven HCL cases. PMID- 2993471 TI - High affinity binding of 125I-labeled human tumor necrosis factor (LuKII) to specific cell surface receptors. AB - 125I-labeled TNF(LuKII) (tumor necrosis factor) binds specifically to human and mouse cell lines sensitive to the cytotoxic effect of TNF, but not to cells made resistant to TNF. TNF-sensitive cells have cell surface receptors with a high affinity for TNF(LuKII). Mouse TNF competes with TNF(LuKII) for receptor binding. Scatchard analysis of the binding data yielded linear plots and suggests that TNF(LuKII) binds to homogeneous receptor sites. The number of TNF(LuKII) receptors on two TNF-sensitive cell lines is 200-300 per cell and the affinity constant of the receptor for TNF(LuKII) is approximately 1 X 10(-10) M. PMID- 2993473 TI - Cannabis: pharmacology and interpretation of effects. AB - A selective introductory review of the Cannabis literature is presented. Subjects reviewed include the relative psychoactivities of Cannabis constituents, the disposition and distribution of THC and its metabolites, the relative psychoactivities of THC metabolites, and the use of cannabinoid concentrations in physiological fluids in interpretations of the significance of Cannabis-induced effects. The pharmacology of cannabinoids in humans is emphasized. PMID- 2993472 TI - Improving physician compliance with preventive medicine guidelines. PMID- 2993474 TI - An evaluation of fused silica capillary columns for the screening of basic drugs in postmortem blood: qualitative and quantitative analysis. AB - Fused silica capillary columns (Durabond) have been evaluated for the screening of more than 100 basic drugs in postmortem blood samples. The combination of these columns, nitrogen-phosphorus detectors, and SKF-525A (internal standard) allows for the simultaneous screening and quantitation of several basic drugs such as amphetamines, amitriptyline, and codeine. Approximately 2000 blood samples have been analyzed by this procedure. The use of capillary columns results in excellent baseline stability and this, together with an autosampler and data system, enables unattended overnight operation. "Double peaking" associated with splitless injection can be a problem as can sensitivity for some of the polar drugs; however, with the extraction procedure described and the equipment used, the screening of blood for basic drugs is improved when compared with packed column technology. PMID- 2993475 TI - Simultaneous electrophoretic determination of phosphoglucomutase subtypes, adenosine deaminase, erythrocyte acid phosphatase, and adenylate kinase enzyme phenotypes. AB - Many of the conventional agarose phosphoglucomutase (PGM) subtyping systems presently in use fail to provide a good separation between the 1 + and 2- bands as well as the 2+ band and the more anodic moving bands. Use of a 1-mm-thick gel composed of 1% ISO GEL (FMC Corp.) and phosphate-citric acid gel and tank buffers with a pH of 5.3 provided exceptionally good separation between all four of the major subtyping bands. The additional criteria for this procedure is a voltage of 21 V/cm and a run time of 4 h. Utilization of this procedure using case samples of varied ages proved the reliability of the procedure. Also examined were the effects of several reducing agents on the enzyme band patterns and the use of this system for the simultaneous determinations of the adenosine deaminase (ADA), erythrocyte acid phosphatase (EAP), and adenylate kinase (AK) enzyme phenotypes. PMID- 2993476 TI - Neurophysiological and biophysical evidence on the mechanism of electric taste. AB - The phenomenon of electric taste was investigated by recording from the chorda tympani nerve of the rat in response to both electrical and chemical stimulations of the tongue with electrolytes in order to gain some insight into its mechanism on both a neurophysiological and biophysical basis. The maximum neural response levels were identical for an individual salt (LiCl, NaCl, KCl, or CaCl2), whether it was presented as a chemical solution or as an anodal stimulus through a subthreshold solution. These observations support the idea that stimulation occurs by iontophoresis of ions to the receptors at these current densities (less than 100 microA/cm2). Electric responses through dilute HCl were smaller than the chemically applied stimulations, but the integrated anodal responses appeared similar to chemical acid responses, as evidenced by an OFF response to both forms of stimuli. Hydrogen may be more permeant to the lingual epithelium and would thus be shunted away from the taste receptors during anodal stimulation. When the anion of electric taste was varied via subthreshold salt solutions, the response magnitude increased as the mobility of the anion decreased. The transport numbers of the salts involved adequately explains these differences. The physical aspects of ion migration occurring within the adapting fluid on the tongue are also discussed. Direct neural stimulation by the current appears to occur only at higher current densities (greater than 300 microA/cm2). If the taste cells of the tongue were inactivated with either iodoacetic acid (IAA) or N-ethyl maleimide (NEM), or removed with collagenase, then responses from the chorda tympani could be obtained only at these higher current densities. Latency measurements before and after IAA or NEM treatment corroborated these findings. The results are discussed in terms of several proposed mechanisms of electric taste and it is concluded that an ion accumulation mechanism can adequately explain the data. PMID- 2993477 TI - The ts110 Moloney mouse sarcoma virus system: gag-mos gene products and cellular transformation. PMID- 2993478 TI - Monoclonal antibody detects a common surface antigen on two independently established murine Moloney sarcoma virus non-producer transformants. AB - Monoclonal antibodies were raised against a cell surface glycoprotein (46K antigen) expressed by a Moloney sarcoma virus (Mo-MSV)-transformed STU mouse cell line (Sac). Although non-productively transformed Sac cells expressed the 46K antigen at their surface, transplantation of Sac non-producer cells into immunocompetent mice failed to induce anti-46K antibody production. Only transplantation of helper virus-superinfected Sac cells led to the development of anti-46K antibodies in addition to antibodies against helper virus structural antigens. Hybridomas producing anti-46K antibodies were established by fusion of mouse myeloma cells with spleen cells taken from mice bearing Sac tumour cells infected with Moloney helper virus. The monoclonal antibodies were subsequently tested against STU mouse transformants [spontaneously transformed or experimentally transformed with the progressor strain of Mo-MSV and the chemical carcinogen 3-methylcholanthrene (MCA), respectively] and BALB/c mouse transformants [transformed with Mo-MSV, simian virus 40 (SV40) or MCA]. Only one of the transformed cell lines tested, a non-producer transformant (PV-TC-77) induced with the progressor strain of Mo-MSV in an STU mouse was found to express a 46K antigen on the cell surface. The 46K surface antigen expressed by PV-TC-77 cells exhibited the same serological, biochemical and biological properties as the 46K Sac cell surface antigen. Though detectable on the surface of PV-TC-77 non-producer cells, immunocompetent syngeneic recipients of non-producer PV-TC-77 cells remained unresponsive against their 46K surface antigen. Only transplantation of helper virus-infected PV-TC-77 producer cells led to antibody development against the 46K surface antigen. PMID- 2993479 TI - An adenovirus cytocidal function related to the control of a cellular pH 4 endonuclease activity. AB - An adenovirus (Ad) interserotypic recombinant (H2cyt141) between temperature sensitive mutant H2ts111 of Ad2 and deletion mutant H5dl313 of Ad5 was isolated and characterized. It was phenotypically ts+, dl+, hr+ and formed large plaques (or cytocidal: cyt). It contained the right 89% of Ad5 DNA and the leftmost 11% of Ad2 DNA. Genetic recombination data suggested the cytocidal mutation lay in the transforming region E1B, confirming sequence analysis. The cytocidal effect resulted in part from the breakdown of cellular DNA. Host cell and virus DNA breakdown induced by H2cyt141 appeared cell-dependent: it occurred in HeLa, KB or BHK-21 cells, but not in CV1 or 293 cells. In human cells the cyt effect was recessive and adenovirus DNA degradation was prevented by co-infection with adenovirus wild-type (H2WT), other adenovirus serotypes or simian virus 40 (SV40). In simian cells, H2cyt141 did not inhibit SV40 DNA replication, unlike H2WT. The amount of H2cyt141 DNA integrated in human cell DNA at early stages of the lytic cycle was found to be significantly lower than for H2WT. Novobiocin inhibited viral DNA breakdown in human cells. Cellular DNA extracted from H2cyt141-infected cells exhibited a repeat band pattern in gel electrophoresis reminiscent of the nuclease digestion pattern of chromatin, with monosome-size fragments as the digestion limit. The H2cyt141-induced nucleolytic effect would therefore occur in the linker regions of cell DNA and might result from the observed stimulation (by a factor of greater than 100) of an acidic (optimum pH 4.0) endonuclease activity. The nucleolytic effect also appeared to be recessive in vitro and absent in mixed samples containing extracts from H2cyt141-infected cells plus extracts from H2WT- or mock-infected cells. The virus gene product responsible for the enhancement of the acidic endonuclease was found to function stoichiometrically and not catalytically. The cytocidal and nucleolytic effects of the viral E1B region 19K protein may be mediated by a cellular inhibitor of acidic endonuclease. PMID- 2993480 TI - Biochemical evidence for the oligomeric (possibly trimeric) structure of the major inner capsid polypeptide (45K) of rotaviruses. AB - The major structural polypeptide of rotaviruses is p45K (VP6), which forms the morphological subunit of the inner capsid. Such subunits show a trimeric structure when examined with the electron microscope. Treatment of single-capsid rotavirus particles with 1.5 M-CaCl2 removes p45K, resulting in the generation of smooth cores. Sucrose density gradient centrifugation analysis of the removed p45K revealed that it has a sedimentation coefficient close to 7.3S, compatible with an oligomeric (possibly trimeric) structure. Polyacrylamide gel electrophoresis under reducing or non-reducing conditions indicated that p45K has intramolecular but not intermolecular disulphide bonds, suggesting that interactions between p45K monomers may be due to some other type of association, such as hydrophobic or charge interactions. Velocity sedimentation of infected cell extracts revealed that native p45K also behaves as an oligomeric protein. Such results were confirmed using p45K partially purified by DEAE-cellulose chromatography. The evidence obtained indicated that all p45K present in the virion is in the oligomeric form, not associated by disulphide bonding, and that most native p45K present in the infected cells is also in the oligomeric form, probably as a consequence of early protein-protein interaction in rotavirus morphogenesis. PMID- 2993481 TI - The coexistence of multiple bluetongue virus electropherotypes in individual cattle during natural infection. AB - Evidence of multiple genotypes of bluetongue virus (BTV) serotype 11 simultaneously infecting individual cattle was demonstrated in a California sentinel herd naturally infected with two BTV serotypes (11 and 17). Monitoring of weekly virus isolates by PAGE demonstrated genome segment diversity among BTV serotype 11 isolates and an individual bull simultaneously infected with multiple electropherotypes of this serotype. No electrophoretic variations were apparent in BTV serotype 17 isolates. The results of this study indicate that multiple plaque clonings and comparative electrophoresis are required to analyse BTV field isolates accurately for the presence of mixed infections. PMID- 2993482 TI - Transmissible gastroenteritis (TGE) of swine: survivor selection of TGE virus mutants in stomach juice of adult pigs. AB - Two transmissible gastroenteritis (TGE) virus mutants (188-SG and 152-SG) were obtained from a low-passage virus strain (D-52) by 188 and 152 cycles of stomach juice treatment and multiplication in cell culture. Compared to the high-passage Purdue-115 and the original D-52 strains, these mutants were more stable at pH 2.0, more resistant to pepsin and trypsin, and characterized by a small plaque phenotype. In vivo, the two mutants were not found to be virulent for 4-day-old piglets and sows after oral inoculation. To test induction of lactogenic immunity, the 188-SG mutant was administered orally to pregnant sows (6 or 7 weeks before parturition) followed by one intramuscular booster (1 week before parturition). After challenge with virulent TGE virus, piglet mortality 7 days after exposure was reduced (to 22%) as compared to the death rate in piglets from control sows (91%). PMID- 2993483 TI - Analysis of the secondary structure of the poly(C) tract in foot-and-mouth disease virus RNAs. AB - Sodium bisulphite modification of foot-and-mouth disease virus (FMDV) RNA in solution indicates that the majority of the poly(C) tract in the RNA is single stranded in concordance with previous results with encephalomyocarditis virus RNA. The reaction kinetics are biphasic; 60% of the cytidylic acid in the poly(C) tract reacts like synthetic poly(C), and the remainder with the kinetics of the cytidylic acid in the rest of the RNA. The reactivity of the poly(C) tract with poly(I) indicates that it is looped out and exposed in the RNA. The deamination reaction has also been used to investigate the structure of the replicative form (RF) and replicative intermediate (RI) isolated from infected cells. Analysis by gel electrophoresis of the long RNase A- and T1-resistant oligonucleotides of RI suggests that it has five single-stranded poly(C) tracts to every one which is base-paired. Bisulphite reactivity of the poly(C) tract and gel electrophoresis of the ribonuclease-resistant oligonucleotides of RF indicate that the poly(C) is base-paired to a poly(G) tract in this molecule. The presence of a poly(G) tract in RF and RI provides unequivocal evidence that the poly(C) is replicated via poly(G) in the negative strand. PMID- 2993484 TI - Chromatin structure of Epstein-Barr virus. AB - The episomal copies of Epstein-Barr virus (EBV) DNA are in a chromatin structure in the lymphoblastoid cell line B95-8. Nucleosomes on EBV DNA have the same spacing as those in cellular chromatin. Some of the EBV DNA is not in nucleosomes; this probably corresponds to DNA which is being packaged. Several DNase hypersensitive sites have been mapped on the EBV genome and patterns of CpG methylation are examined. PMID- 2993485 TI - Studies on the pathogenesis of a bovine cytomegalo-like virus in an experimental host. AB - Herpesvirus Movar 33/63 is the prototype strain of a group of slow growth bovine herpesviruses which have been reported to exhibit cytomegalovirus-like characteristics. These viruses have the ability to produce long-term persistent infections of spleen and other lymphoreticular organs in both cattle and rabbits. Rabbits were inoculated with a suspension of Movar 33/63 propagated in cell culture, and sacrificed at intervals between 3 days and 49 weeks post-infection. Cell-free infectious virus was detected only in conjunctival secretions, buffy coat and spleen homogenates up to 7 days post-infection. Beyond this brief acute replication period, co-cultivation or explantation was required for the detection of viral infectivity. The spleen, the only organ from which virus was consistently recovered, exhibited the highest infectious titres as detected by infectious centre assay. The use of several cell isolation techniques (including solid-phase fractionation on ligand-coated surfaces, nylon wool filtration, affinity chromatography, immunocytolysis and plastic surface adherence) allowed separation of B-enriched, T-enriched and non-T, non-B cell fractions. Infectivity during the acute and persistent phase of the infection was associated with the non-T, non-B population which was highly enriched in adherent and non-adherent spleen mononuclear phagocytes. No virus was isolated from either T or B cells. PMID- 2993486 TI - The therapeutic efficacy of a xanthate compound on herpes simplex virus in skin lesions of mice and guinea-pigs. AB - Xanthates have recently been shown to inhibit the replication of both DNA and RNA viruses in vitro. The antiviral activity was exerted only under acidic pH conditions. Curative effects in vivo on herpes simplex virus (HSV)-induced skin lesions were only observed when the xanthate compound was administered in the form of an ointment containing acidic buffer (sodium phosphate pH 5.0). Advanced HSV-2-induced skin lesions in mice were healed by topical treatment with the xanthate compound. HSV-1-induced lesions on skin of guinea-pigs were cured within 2 days even when the treatment was initiated as late as 4 days after infection. Both HSV-1 DNA synthesis and virus production in the skin of guinea-pigs were also shown to be inhibited after treatment with the xanthate compound. PMID- 2993487 TI - Transcription of the Bacillus subtilis bacteriophage phi 3T in vitro. AB - The in vitro transcription pattern of Bg/II-digested phi 3T DNA is described. Eight Bg/II fragments that hybridized to in vitro transcription products were unequivocally identified. A further hybridizing region corresponding to a Bg/II triplet was also revealed, giving a total of nine to 11 Bg/II fragments. These represent 47 to 53% of the phi 3T genome. Transcription was shown to initiate within Bg/II fragments B, G, C, H, I, F and J, implying that all of these contain at least one promoter. The relevance of these data to the construction of a cloning vector based on phi 3T is discussed. PMID- 2993489 TI - Monoclonal antibodies recognizing early and late antigens of human cytomegalovirus: heterogeneity of polypeptides recognized between virus isolates. AB - The characteristics of four human cytomegalovirus (HCMV)-specific monoclonal antibodies as assessed by ELISA, immunofluorescence, immunoprecipitation and Western blotting are described. Two antibodies recognized a 67K late polypeptide of HCMV, one recognized 43K and 79K polypeptides present early and late in HCMV infected cells, and the fourth identified a 72K early nuclear protein of HCMV. The antibodies recognized these antigens in all HCMV isolates tested by immunofluorescence and ELISA, but demonstrated inter-isolate variations in polypeptides recognized by Western blotting. PMID- 2993490 TI - Immunological and biochemical characterization of HZ2 feline sarcoma virus and Abelson murine leukaemia virus translation products. AB - The extent of homology between the translation products of the HZ2 strain of feline sarcoma virus (HZ2-FeSV) and the Abelson murine leukaemia virus (A-MuLV) was examined immunologically and biochemically. Antiserum prepared against the v abl-encoded determinants of the A-MuLV polyprotein P120gag-abl was also found to precipitate specifically the 98K mol. wt. HZ2-FeSV protein (P98gag-abl). The basis for this immunological crossreactivity was indicated by the findings that the two proteins had at least six [35S]methionine-containing tryptic peptides and at least eight [35S]methionine-containing chymotryptic peptides in common. Each of the two proteins also had tryptic and chymotryptic peptides which were unique. Both proteins were associated with tyrosyl kinase activities which exhibited some similar biochemical properties in vitro. However, the HZ2-FeSV-associated activity was much more sensitive to competitive inhibition by nucleoside and deoxynucleoside diphosphates than was the A-MuLV-associated activity. These results suggest that, while the gag-abl translation products of these two independent isolates of transforming retrovirus are highly related structurally and functionally, the differences in structure contribute to differences in enzyme activity. Further comparative studies of these two proteins should play an important role in determining their roles in induction of two different types of malignancy: lymphosarcoma in the case of the A-MuLV protein and fibrosarcoma in the case of the HZ2-FeSV protein. PMID- 2993488 TI - Identification of cross-reacting glycoproteins of four herpesviruses by Western blotting. AB - Monospecific rabbit antisera against purified herpes simplex virus type 1 (HSV-1) gB, gC or gD antigens or polyvalent rabbit antiserum against equine herpesvirus type 1 (EHV-1)-infected cells was used in Western blotting to identify antigenically related proteins in cells infected with HSV-1, HSV-2, bovine mammillitis virus or EHV-1. Monomeric and oligomeric polypeptides related to HSV 1 gB were found in cells infected with each of the four herpesviruses. The gC antiserum was specific for HSV-1 and the gD antiserum reacted only with polypeptides in HSV-1- or HSV-2-infected cells. Neither gC nor gD antiserum revealed the existence of oligomeric forms of the glycoproteins with the single exception of HSV-1 strain KOS which had a heat-unstable high molecular weight form gD. The EHV-1 antiserum reacted with high molecular weight polypeptides of the same mobility as the oligomeric form of gB in cells infected with each of the four herpesviruses. PMID- 2993491 TI - Enhanced intranuclear expression of measles virus following exposure of persistently infected cells to cyclic AMP. AB - The intracellular distribution of measles virus inclusion bodies in persistently infected human cells (AV3A1/MV) changed markedly following continuous exposure to 3', 5' cyclic adenosine monophosphate (cAMP). When assayed by immunofluorescence, the number of cells with intranuclear virus inclusions increased from 5 to 10% to 80 to 90% after exposure to 1 mM-cAMP for 4 days. Exposure of cells to cAMP also resulted in a twofold increase in the average number of inclusions in invaded nuclei. Similar but less pronounced changes occurred in cells treated with inducers of adenylate cyclase and an inhibitor of phosphodiesterase. Examination of cAMP-treated cells by electron microscopy indicated that viral inclusion bodies consisted of typical helical nucleocapsids. No evidence of nucleocapsids crossing the nuclear membrane (through nuclear pores) was found. PMID- 2993492 TI - Detection of adenoviruses in stool specimens by nucleic acid spot hybridization. AB - Nucleic acid hybridization was used for the detection of adenovirus DNA in stool specimens, and the results were compared with those obtained by a radioimmunoassay (RIA) for adenovirus hexon antigen. DNA from 40 specimens, 18 of which were positive by RIA, were spotted onto nitrocellulose filters and analyzed by hybridization using radioactively labeled adenovirus-2 DNA or a cloned DNA fragment from enteric adenovirus-41 as probes. With the adenovirus-2 DNA probe, 15 of the 18 RIA-positive specimens were also positive in the hybridization assay, and one of the RIA negative specimens was also scored as positive. The cloned adenovirus-41 fragment gave a positive signal with five specimens, all of which were also detected with the adenovirus-2 DNA probe. The results show that hybridization is an alternative method for detection of adenovirus in stool specimens. The sensitivity of the assay is comparable to that of the RIA. PMID- 2993493 TI - Comparison of various serological methods and diagnostic kits for the detection of acute, recent, and previous rubella infection, vaccination, and congenital infections. AB - The antibody development after natural rubella infection and rubella vaccination has been followed in 802 sera from 493 patients and 71 sera from 22 vaccinees. Also examined were 67 sera from 28 infants with rubella embryopathy and sera from 50 children with presumed prenatal infection. In addition, 777 sera from 641 patients tested for routine rubella diagnosis were studied. Anamnestic information was available from all these patients. These sera were assayed for IgM antibody detection by sucrose density gradient (SDG), the commercial ELISAs (Enzygnost IgM and Rubazyme M), and the non-commercial anti-my-hemadsorption immunosorbent technique (HIT). For the determination of IgG antibodies the hemagglutination inhibition test (HAI), the commercial ELISAs (Enzygnost IgG, Rubazyme), and a single radial hemolysis test (SRH) were used. The SDG and HIT were less sensitive for IgM antibody detection than the two ELISAs, particularly when IgM concentrations were low. In total 26.5% of the IgM results with the newer tests were discordant with SDG, but only 0.5-1.3% of these results were not explicable when the clinical data was considered. Problems were encountered with all IgM assay systems used. For the detection of rubella antibodies after acute infection and vaccination the ELISA Enzygnost IgG was as sensitive as the HAI whereas the ELISA Rubazyme and SRH detected antibodies with some delay. Corresponding results with all tests were found more than 25 days after acute infection and more than 50 days after vaccination. All methods can be used for detection of antibodies in infants with rubella embryopathy. The results of this study suggest that certain combinations of tests can be used for the reliable detection of rubella infection. PMID- 2993494 TI - Natural killing (NK) of cytomegalovirus (CMV)-infected fibroblasts: a comparison between two strains of CMV, uninfected fibroblasts, and K562 cells. AB - Spontaneous cytolysis of uninfected human fibroblasts (HF), K562 cells, and HF cells infected with cytomegalovirus (CMV) was associated with nonadherent peripheral blood leukocytes (PBL) that passed through a nylon wool column, and rosetted with sheep erythrocytes at low affinity; cytolysis was further associated with enriched preparations of large granular lymphocytes (LGL). The Leu-7 marker did not correlate as a cell phenotype with functional activity in normal donors. The HF cells infected with the AD169 strain of CMV were as sensitive to cytolysis as K562 cells, although the kinetics of cytolysis differed; HF cells infected with the Davis strain of CMV were generally less sensitive to cytolysis than uninfected fibroblasts. Equivalent amounts of interferon were detected in NK assays containing Davis targets, AD169 targets, or K562 cells; interferon was not detected in NK assays of HF cells. Neither uninfected nor infected HF cells competed effectively against 51Cr-labeled K562 cells in cold competition experiments; K562 cells, however, competed effectively against 51Cr-labeled AD169-CMV targets. Uninfected HF cells were efficient competitors of AD169-CMV targets. Davis-CMV-infected cells were less effective, however, than uninfected HF cells in competition experiments, suggesting that fewer receptors for NK cells were present on these targets. PMID- 2993495 TI - Serum antibodies to the major HSV-2-specified DNA-binding protein in patients with an acute HSV infection or cervical neoplasia. AB - The major HSV-2-specified DNA-binding protein (ICSP 11/12) was purified from HSV 2-infected cells. ELISA and immunoblotting techniques were used to study its antigenicity in HSV-infected patients and patients with cervical neoplasia and control women. Patients with an acute HSV-2 infection had clearcut antibody responses to the purified ICSP 11/12 preparation. Determination of the ICSP 11/12 antibodies by ELISA revealed considerably higher serum antibody levels in patients with cervical carcinoma than in the controls. PMID- 2993497 TI - Inhibition of in vitro HBsAg production by amphotericin B and ketoconazole. AB - The effects of amphotericin B, ketoconazole, and adenine arabinoside on production of hepatitis B surface antigen (HBsAg) particles by the human hepatoma cell line PLC/PRF/5 were examined. In addition, the effects of these drugs on cellular protein synthesis were determined. These drugs caused a dose-dependent decrease in HBsAg production that was paralleled by a decrease in cellular protein synthesis. Ketoconazole was the most active of these drugs and the most specific, causing a 72% reduction in HBsAg production with only a 38% reduction in protein synthesis. These data suggest that further studies evaluating ketoconazole for the treatment of chronic hepatitis B virus infection in animals are warranted. PMID- 2993496 TI - Detection and localization of human papillomavirus DNA in human genital condylomas by in situ hybridization with biotinylated probes. AB - We have examined the distribution of human papillomavirus (HPV) DNA in paraffin sections of humans warts by in situ hybridization with biotin-labeled DNA probes. Recombinant plasmid DNAs (HPV-1, -6, -11, -16) were labeled by nick translation with biotinylated deoxyuridine triphosphate. Paraffin sections were hybridized with the probes for 18 h in stringent or non-stringent conditions, and DNA-DNA hybrids were detected by immunocytochemistry. Paraffin sections of warts were also examined for the presence of HPV capsid antigen with the avidin-biotin peroxidase complex method for immunocytochemistry. HPV DNA was detected and localized in paraffin sections from a plantar wart, a laryngeal papilloma, and seven anogenital condylomas. The specific HPV type present in each lesion was determined by hybridization under stringent conditions with the homologous DNA probe. The papillomas were found to contain many more cells with replicating virus DNA, as demonstrated by in situ hybridization, than was apparent from the number of cells containing detectable virus antigen. In situ hybridization with biotin-labeled probes is an effective technique for the identification of HPV infection in routinely collected and processed tissue specimens. PMID- 2993498 TI - Lack of oral HSV-2 in a college student population. AB - A population of individuals with a high incidence of genital herpes simplex virus type 1 (HSV-1), due most likely to oro-genital contact, was examined to determine the incidence of oral herpes simplex virus type 2 (HSV-2) infection. Herpes simplex virus was isolated from the oral cavity of 43 college students whose symptoms ranged from singular lesions of the lips with minimal discomfort to severe oral disease with systemic involvement resulting in lymphadenopathy, chills, sweat, myalgia, and fever. The virus isolated from each case was identified by serum neutralization and typed as HSV-1 or HSV-2 using (E)-5-(2 bromovinyl)-2'-deoxyuridine (BVDU) sensitivity, monoclonal antibody immunofluorescence, and restriction endonuclease EcoRI digestion of viral DNA. In every instance the isolate was HSV-1. Additional identification and typing of head and neck isolates as well as oral samples from non-university patients demonstrated that all were also HSV-1. Therefore, while HSV-1 appears to be readily transmitted to the genitalia in this group of individuals, the transmission of HSV-2 to the oral cavity may not be as common, even though clinical histories revealed that several of these patients were engaging in oro genital contact. PMID- 2993499 TI - Immunostimulatory function of herpes simplex virus isolates from patients with frequent herpes labialis and a deficiency in immune-specific interferon production. AB - Approximately 30% of persons with frequent episodes of herpes labialis are deficient in the production of HSV-induced immune-specific interferon (IFN) (Green, 1985). Herpes simplex virus (HSV) strains isolated from persons who make immune-specific IFN and from persons who do not make it were examined for their immunostimulatory capabilities. HSV isolated from the primary oral lesions of two patients deficient in immune-specific IFN production, one person with an intact immune-specific IFN response, HSV types 1 and 2 laboratory strains, and Newcastle disease virus (NDV) were added to cultures of peripheral blood mononuclear cells (PBML) from HSV seropositive donors. All HSV-isolates induced comparable titers of immune-specific IFN. These studies suggest that failure of some patients to develop an immune-specific IFN response is determined by the host, not the virus. PMID- 2993500 TI - Immune-specific interferon production by peripheral blood mononuclear leukocytes from patients with primary and recurrent oro-labial herpes simplex virus infections. AB - Immune-specific IFN (IFN) is produced by the peripheral blood mononuclear leukocytes (PBML) of greater than 95% of HSV-seropositive humans with infrequent recurrences of herpes labialis [Green, Yeh, and Overall, 1981]. However, herpes virus-induced immune-specific IFN was produced by PBML from only 33 of 48 (68.8%) persons with frequent recurrences (2-12 episodes a year). Two of eight subjects with primary herpes gingivostomatitis also failed to produce immune-specific IFN during either the acute or convalescent phases of their initial HSV infection. These data suggest that some persons have a defective immune-specific IFN response that exists from the time of their primary oro-labial HSV type 1 infection. This defect may predispose to a higher frequency of disease in some individuals. PMID- 2993501 TI - Detection of markers of hepatitis B virus infection in urine of chronic carriers. AB - Urine from 19 chronic hepatitis B virus (HBV) carriers was tested for markers of HBV infection. Hepatitis B surface and e antigens were excreted in the urine of about one-half of the patients, but not in any consistent pattern. HBV DNA, a much more sensitive indicator of infectivity, was found in only one specimen and then in very low concentration. We conclude that urine of chronic HBV carriers is not a major vehicle for the transmission of hepatitis B. PMID- 2993503 TI - Detection of BK virus antibodies measured by enzyme-linked immunosorbent assay (ELISA) and two haemagglutination inhibition methods: a comparative study. AB - We developed a solid phase enzyme-linked immunosorbent assay (ELISA) for detecting IgG antibodies to BK virus (BKV). This test was compared to two haemagglutination inhibition methods, one using receptor-destroying enzyme and the other NaIO4 to remove nonspecific inhibitors. The results show that this single dilution ELISA is a sensitive and specific test. The inherent advantages of this method make it an attractive alternative for the seroepidemiology of BKV. PMID- 2993502 TI - Persistence of serum IgA antibodies to herpes simplex, varicella-zoster, cytomegalovirus, and rubella virus detected by enzyme-linked immunosorbent assays. AB - Enzyme-linked immunosorbent assays were used to detect IgG and IgA antibodies to herpes simplex virus (HSV), varicella-zoster virus (VZV), cytomegalovirus (CMV), and rubella virus in sera from 68 adult female gynaecological patients. Of the patients who had virus-specific IgG antibodies, the proportion who also had virus specific IgA was 98% for HSV, 75% for VZV, 73% for rubella virus, and 42% for CMV. IgA antibodies to all four viruses were only found when specific IgG antibodies were also detected in the serum. These results suggest that virus specific IgA may persist for several years; possible explanations for this are discussed. PMID- 2993504 TI - Virological and serological diagnosis of cytomegalovirus infection in bone marrow allograft recipients. AB - To detect cytomegalovirus (CMV) infections, a total of 1,074 cultures of urine, saliva, or blood were collected weekly from 43 consecutive patients undergoing allogeneic bone marrow transplantation. Twenty-three patients were seronegative before transplant and primary infection occurred in 2 (9%). Twenty patients were initially seropositive and recurrent infections occurred in 5 (25%). Three patients in the recurrent group had proven CMV pneumonitis; viraemia was detected in two recipients, while the third had CMV isolated only from bronchial lavage fluid. The serological response of the 43 patients was defined by testing 559 serial sera for specific IgG and IgM antibodies by radioimmunoassay. Passive acquisition of IgG antibodies from blood products was found in 78% of initially seronegative recipients. One patient with primary infection responded in a pattern typical of immunocompetent individuals with long-term production of specific IgG and transient production of specific IgM antibodies. The second patient also had a typical response, but this was delayed until several weeks after the start of virus excretion. In patients with recurrent infections, specific IgM production did not correlate with episodes of virus excretion. Three of five such patients failed to mount a specific IgM response, and these were the only patients in the study to develop CMV pneumonitis. We conclude that CMV infection in bone marrow recipients can only be diagnosed by detection of virus; therefore, the ability of these patients to mount humoral immune responses should not be relied upon for diagnostic purposes. PMID- 2993505 TI - Detection of cytomegalovirus in urine samples by enzyme-linked immunosorbent assay. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of cytomegalovirus (CMV) in urine using monoclonal antibodies directed against CMV as a capture for viral antigen. The assay was capable of detecting virus at 10(2.3)TCID50/ml as determined by titration of stock virus, strain Ad169. The assay was found to have a sensitivity of 65% and a specificity of 100% when 73 coded stored urine specimens were examined. Assuming that the poor sensitivity was due to loss of antigen following storage, we proceeded to analyse fresh urine specimens. Surprisingly, the assay gave negative results with 46 fresh urines known to contain CMV; however, following storage at +4 degrees C for two weeks, 35 (76%) of these samples gave ELISA results in the positive range. This detection of CMV, after storage at +4 degrees C, could be due to degradation of virus particles leading to release of soluble glycoproteins into the medium or to the presence of an inhibitory substance in fresh urine that is destroyed during storage. PMID- 2993506 TI - Quantitative and qualitative detection of cytomegalovirus-specific antibodies using two types of enzyme-linked immunosorbent assay. AB - An indirect ELISA and an inhibition ELISA were developed for the detection of cytomegalovirus (CMV)-specific immunoglobulin G (IgG) and CMV-specific total immunoglobulin, respectively. Both assays were more specific than the complement fixation (CF) test, and titres of positive sera were 660 times higher by IgG ELISA and 6 times higher by inhibition ELISA than titres by the CF test. Titres by IgG ELISA were reliably determined using the absorbance obtained at a single serum dilution of 1/1,000 in conjunction with a standard graph. Both ELISAs compared favourably with each other in sensitivity and specificity in determining CMV immune status. The inhibition ELISA, in particular, provides a simple and reliable method of screening sera, which requires no control antigen or predilution of sera. It should prove useful for large-scale screening procedures, such as blood donor testing. PMID- 2993507 TI - Hypothalamic monoaminergic mechanisms of aspirin-induced analgesia in monkeys. AB - Microinjection of sodium acetylsalicylate (aspirin, 0.2-0.6 mg) into the preoptic anterior hypothalamic area, but not the dorsal raphe region or the periaqueductal central gray matter, produced a dose-related analgesia in conscious monkeys. The analgesia induced by intrahypothalamic administration of aspirin was antagonized by pretreatment of monkeys with either a serotoninergic receptor blocker (cyproheptadine) or two catecholaminergic receptor blockers (haloperidol and yohimbine). These suggest the existence of a monoaminergic pain-inhibitory mechanism in the preoptic anterior hypothalamic area activated by aspirin. PMID- 2993508 TI - Interaction between alpha- and beta-adrenoceptors in rat pineal adenosine cyclic 3',5'-monophosphate phosphodiesterase activation. AB - The adrenergic regulation of the low-Km pineal cAMP phosphodiesterase (PDE) activity was studied in adult female rats. PDE activity showed a transient enhancement (up to 42%) during the process of degeneration of pineal sympathetic nerve terminals that followed superior cervical ganglionectomy (SCGx), thus confirming the neural modulation of the enzyme. Treatment with isoproterenol (0.3 5.0 mg/Kg) increased significantly PDE activity within 2 hours. Phenylephrine induced a significant increase of pineal PDE only at a 10 mg/Kg dose, while at a lower dose (1 mg/Kg) it potentiated the stimulatory effect of isoproterenol. Treatment of pineal organ cultures with 100 microM propranolol inhibited norepinephrine (NE)-induced PDE activity while 100 microM phentolamine had no significant effect. Propranolol at doses unable to alter the in vitro NE-induced stimulation of pineal PDE activity (1 microM), antagonized such NE effect when used in combination with 1 microM phentolamine. At equimolecular concentrations (1 microM) the mixed alpha-beta-adrenergic agonist NE was more effective than the beta-adrenergic agonist isoproterenol to increase pineal PDE in vitro. These results suggest an alpha-beta-adrenergic interaction in the sympathetic modulation of low-Km PDE activity of rat pineal gland. PMID- 2993509 TI - Interaction of opiates with opioid binding sites in the bovine adrenal medulla: I. Interaction with delta and mu sites. AB - In the present study we examined the interaction of opiates with the delta and mu opioid binding sites in the bovine adrenal medulla. [3H][D-Ala2, D-Leu5] enkephalin ( [3H]DADLE) in the presence of saturating concentrations of morphiceptin was used to analyze delta site interactions, whereas either [3H]DADLE in the presence of saturation concentrations of [D-Ser2, Leu5] enkephalin-Thr6 (DSLET) or [3H][D-Ala2, Me-Phe4, Gly5-ol]-enkephalin ( [3H]DAGO) was used for the determination of mu sites. Both binding sites were found to interact stereoselectively with opiates. The binding was affected differentially by proteolytic enzymes (trypsin, alpha-chymotrypsin, pepsin), N-ethylmaleimide, and A2-phospholipase. Kinetic and equilibrium binding studies revealed that in each case radiolabeled opiates interact with one class of binding sites, following simple second-order bimolecular kinetics. Competition for binding by opiates and opioid peptides confirmed the delta and mu selectivity of these sites. Monovalent (Na+, Li+, K+) and divalent (Mg2+, Mn2+, Ca2+) ions interacted differentially with these two binding sites: In general, monovalent cations affected preferentially the apparent number of binding sites, whereas divalent ions modified the equilibrium dissociation constant. Furthermore, positive or negative cooperativity and an apparent heterogeneity of binding sites were detected under some ionic conditions. PMID- 2993510 TI - Interaction of opiates with opioid binding sites in the bovine adrenal medulla: II. Interaction with kappa sites. AB - In this study we examined the interaction of opiates with kappa binding sites in the bovine adrenal medulla. [3H]Ethylketocyclazocine (EKC), [3H]etorphine, and [3H]bremazocine stereoselective bindings were used to assay these interactions. The kappa sites were found to be heterogeneous: [3H]bremazocine identified with high affinity all subtypes of these sites. [3H]EKC, in the presence of saturating concentrations of [D-Ala2, D-Leu5]-enkephalin (DADLE) (5 microM), was used to identify kappa 1 sites, on which dynorphin A (1-13) bound with high affinity. Either [3H]EKC or [3H]etorphine in the presence of 5 microM DADLE identified the kappa 2 subtype. This subtype was found to interact with beta-endorphin and especially with the octapeptide Met5-enkephalyl-Arg6-Gly7-Leu8. Furthermore, [3H]etorphine identified in the bovine adrenal medulla a third high-affinity component, in the presence of 5 microM DADLE. This residual interaction was found to be equally stereoselective and presenting kappa selectivity. Met5-enkephalyl Arg6-Phe7 interacted preferentially with this site. The three kappa subtypes interacted differentially with monovalent (Na+, K+, and Li+) and divalent (Ca2+, Mg2+, and Mn2+) ions by modification of the apparent concentration of the accessible sites and/or by changes of the apparent KD for radioligands. Modifying agents (proteolytic enzymes, thiol-modifying reagents, and A2-phospholipase) produced different effects on each subtype of the kappa site, suggesting a different protein (or protein-lipid?) composition. PMID- 2993511 TI - Existence of sites for anions and divalent cations in the solubilized gamma aminobutyric acid/benzodiazepine receptor complex. AB - This study evaluated the ability of gamma-aminobutyric acid (GABA), baclofen, monovalent anions, divalent cations, and various combinations thereof to protect solubilized benzodiazepine (BZ) receptors of types 1 and 2, when contained together on the complex, against heat inactivation. Neither anions, cations, nor GABA alone provided significant protection of solubilized BZ receptors against heat, but inclusion of monovalent anions or divalent cations together with 500 microM GABA did afford protection. Monovalent anions combined with GABA (500 microM) provided 50% to full protection. Divalent cations, such as CaCl2 (2.5 mM) or MgCl2 (2.5 mM) in the presence of GABA (500 microM) yielded 45% and 24% protection, respectively. Other divalent cations tested (Zn2+, Hg2+, Co2+, and Ni2+) were poor protectors, even when combined with GABA. Monovalent anions (200 mM NaCl) and divalent cations (5 mM CaCl2) when tested together provided no protection. Similarly, baclofen (the GABA-B agonist) provided no protection, either alone or together with anions or divalent cations. These results indicate that the independent but interacting recognition sites of GABA, BZ, anions, and divalent cations, previously detected in the membrane-bound state, are retained in the solubilized state. PMID- 2993512 TI - Effect of acute hypoxia on ascorbate content of plasma, cerebral cortex, and adrenal gland. AB - Levels of ascorbic acid (AA) in the plasma, brain, and adrenal gland of rats were determined after 15 min of hypoxia (PaO2 less than 25 mm Hg) and following asphyxia. In rabbits, AA plasma levels were followed up to 75 min of reoxygenation following 15 min of hypoxia of the same severity. A significant increase (approximately 70%) in AA levels was found in plasma of rats and rabbits after hypoxia and asphyxia. This increase was found to be transient, with a return to normal levels within 1 h after resumption of normal oxygenation. Pretreatment with dexamethasone reduced the increase in AA level in both rabbits and rats. Adrenalectomy in rats, performed 24 h before the experiment, abolished the response to hypoxia. Ascorbate levels in the cerebral cortex, hypothalamus, and adrenal gland of awake rats subjected to hypoxia or asphyxia were found to be the same as in normoxic rats. Our results suggest that the observed changes in plasma AA levels are probably mediated through adrenocorticotropic hormone and that the adrenal gland is the major source of ascorbate efflux into the circulation during oxygen deprivation. PMID- 2993513 TI - Nature of extrasynaptosomal accumulation of endogenous adenosine evoked by K+ and veratridine. AB - When rat brain synaptosomes were incubated for 10 min at 37 degrees C, basal accumulation of adenosine in the medium was 66 pmol/mg of protein. An elevated K+ level (24 mM) evoked an additional accumulation of 200 pmol/mg of protein, and 50 microM veratridine evoked 583 pmol of adenosine accumulation/mg of protein. K+- and veratridine-evoked accumulation of adenosine did not arise from microsomal or mitochondrial contaminants of the synaptosomal preparation, because purified microsomes and mitochondria did not exhibit evoked accumulation of adenosine in the medium. K+-evoked accumulation of extrasynaptosomal adenosine was Ca2+ dependent, whereas veratridine-evoked accumulation of adenosine was increased in Ca2+-free medium. In the presence of alpha,beta-methylene ADP and GMP, which inhibit ecto-5'-nucleotidase, conversion of added ATP and AMP to adenosine was inhibited by 90% in synaptosomal suspensions. However, inhibition of ecto-5' nucleotidase only reduced basal extrasynaptosomal accumulation of adenosine by 74%, veratridine-evoked accumulation of adenosine by 46%, and K+-evoked accumulation by 33%. Most of the basal accumulation of extrasynaptosomal adenosine appears to be derived from released nucleotide, probably ATP, but about half of the veratridine-evoked accumulation of adenosine and most of the K+ evoked accumulation may arise from adenosine released in its own right, rather than from a released nucleotide. PMID- 2993514 TI - t-[3H]butylbicycloorthobenzoate: new radioligand probe for the gamma-aminobutyric acid-regulated chloride ionophore. AB - t-[3H]Butylbicycloorthobenzoate [( 3H]TBOB; 22 Ci/mmol) was prepared by reductive dechlorination of its 4-chlorophenyl analog with tritium gas. This new radioligand binds reversibly to fresh washed rat brain P2 membranes in 500 mM NaCl plus 50 mM sodium-potassium phosphate buffer (pH 7.4) at 25 degrees C, with 80-90% specific relative to total binding, a KD of 61 +/- 15 nM, and a Bmax of 1.6 +/- 0.5 pmol/mg of protein. [3H]TBOB association with its binding site(s) is monophasic, but its dissociation is biphasic. The binding characteristics of [3H]TBOB are essentially identical to those of t [35S]butylbicyclophosphorothionate [( 35S]TBPS) with respect to pH dependence, stimulation by anions, regional distribution in the brain, and pharmacological profile. Saturation analyses and dissociation studies further indicate that TBOB and TBPS have a common binding site. However, binding of the two radioligands differs in respect to temperature effects. In contrast to [35S]TBPS, which exhibits negligible binding at 0 degrees C, [3H]TBOB binds to rat brain membranes at 0, 25, and 37 degrees C with similar KD values. [3H]TBOB with its long radioactive half-life and temperature-independent KD is a valuable supplement to [35S]TBPS in further biochemical and pharmacological characterization of the gamma-aminobutyric acid receptor-ionophore complex. PMID- 2993515 TI - Corticotropin-peptide regulation of intracellular cyclic AMP production in cortical neurons in primary culture. AB - Previous studies have provided evidence for adrenocorticotropic hormone (ACTH) effects on a wide variety of behaviors. However, the precise sites of action and the mechanisms by which these effects may be mediated have yet to be clearly elucidated. Although ACTH was shown to augment cyclic AMP levels in glial cells isolated from whole brain, other studies found little or no effect of ACTH peptides on cyclic nucleotide metabolism in slices of cerebral cortex or homogenates of whole brain. In the present study, our objective was to determine whether ACTH peptides regulate intracellular cyclic AMP levels in neurons of the cerebral cortex in primary culture. ACTH peptides stimulated cyclic AMP synthesis up to threefold in a dose-dependent manner; stimulation was complete within 5-10 min of exposure to agonists. Neurohormone efficacy was augmented by 0.1 microM forskolin (which was virtually ineffective alone); potency was unaffected. The order of potency (EC50) for increasing intracellular cyclic AMP levels was as follows: ACTH (1-24), ACTH (1-17) (10 nM) greater than alpha-melanocyte stimulating hormone, beta-melanocyte stimulating hormone (alpha-MSH, beta-MSH) (100 nM) greater than ACTH (1-10) (1 microM) greater than ACTH (4-10) (5 microM). The hexapeptide ACTH (4-9) as well as ACTH (11-24) were inactive at concentrations as high as 10 microM. Other neuropeptides derived from proopiocortin, such as beta-endorphin and Met- and Leu-enkephalin were without effect on basal or hormonally stimulated cyclic AMP synthesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993516 TI - Characterization of functional receptors for vasoactive intestinal peptide in bovine cerebral arteries. AB - This study reports the characterization of receptors for vasoactive intestinal peptide (VIP) on membranes prepared from bovine cerebral arteries. By use of HPLC we prepared two purified monoiodinated VIP radioligands with nearly equivalent cerebral vasorelaxant potency as native VIP, [Tyr(125I)10 )VIP and [Tyr(125I)22]VIP. The former resulted in a higher proportion of specific binding to arterial membranes than the latter and was therefore thought to be the superior radioligand for receptor characterization. The binding of [Tyr(125I)10]VIP to cerebral arterial membranes was saturable, specific, reversible, and dependent on time and temperature. Scatchard analysis suggested the presence of a high- and a low-affinity binding site with KD values of 0.2 and 11 nM and receptor concentrations of 79 and 737 fmol/mg of protein, respectively. The dose-response curves for binding to the VIP receptor by the VIP-homologous peptides PHI, PHM, and rat growth hormone-releasing factor (GRF) were very similar to their dose-response curves for relaxation of cerebral arteries. The order of potency was VIP greater than PHM greater than PHI greater than rat GRF. It is suggested that the characteristics of the vascular VIP binding sites and the close correlation between the binding and vasorelaxant properties of VIP and its related peptides argue for the vascular binding sites being functional receptors for VIP. PMID- 2993517 TI - Phosphorylation of microtubule-associated protein 2 at distinct sites by calmodulin-dependent and cyclic-AMP-dependent kinases. AB - Microtubule-associated protein 2 (MAP2) is an excellent substrate for both cyclic AMP (cAMP)-dependent and Ca2+/calmodulin-dependent kinases. A recently purified cytosolic Ca2+/calmodulin-dependent kinase (now designated CaM kinase II) phosphorylates MAP2 as a major substrate. We now report that microtubule associated cAMP-dependent and calmodulin-dependent protein kinases phosphorylate MAP2 on separate sites. Tryptic phosphopeptide digestion and two-dimensional phosphopeptide mapping revealed 11 major peptides phosphorylated by microtubule associated cAMP-dependent kinase and five major peptide species phosphorylated by calmodulin-dependent kinase. All 11 of the cAMP-dependently phosphorylated peptides were phosphorylated on serine residues, whereas four of five major peptides phosphorylated by the calmodulin-dependent kinase were phosphorylated on threonine. Only one peptide spot phosphorylated by both kinases was indistinguishable by both migration and phosphoamino acid site. The results indicate that cAMP-dependent and calmodulin-dependent kinases may regulate microtubule and cytoskeletal dynamics by phosphorylation of MAP2 at distinct sites. PMID- 2993518 TI - Wheat germ agglutinin inhibits the effects of nerve growth factor on the phosphorylation of proteins in PC12h cells. AB - Treatment of PC12h cells in tissue culture with nerve growth factor (NGF) led to an increased incorporation of [32P]orthophosphoric acid into specific proteins. The increased phosphorylation of 60,000-dalton and 20,000-dalton proteins in the 0.2% Triton X-100 detergent-soluble fraction, of 35,000-dalton protein in the 0.2% Triton X-100 detergent-insoluble fraction, and of slow migrating protein (SMP) in the nonhistone nuclear fraction was observed upon NGF treatment. On the other hand, wheat germ agglutinin (WGA) treatment of PC12h cells induced a slightly decreased phosphorylation of these NGF-responsive proteins. Incubation of cell-free extracts from PC12h cells with [gamma-32P]ATP led to the phosphorylation of a 100,000-dalton protein. In extracts from cells treated with NGF, the labeling of the 100,000-dalton protein was substantially and selectively reduced. In contrast, treatment of PC12h cells with WGA led to an increased phosphorylation of the 100,000-dalton protein in cell-free extracts. Thus, NGF and WGA showed opposite effects on the phosphorylation of specific proteins in both intact cells and cell-free extracts. In addition, it was also observed in both systems that pre- and posttreatment of PC12h cells with WGA abolished the effects of NGF on the phosphorylation and produced a phosphorylation pattern similar to that from PC12h cells treated only with WGA. In parent PC12 cells, it has been reported that the treatment of cells with WGA inhibits NGF binding to its receptors and converts the rapidly dissociating receptors to slowly dissociating receptors. Thus, WGA in conjunction with NGF, results in the practical disappearance of rapidly dissociating receptors on cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993519 TI - Dibutyryl-cyclic GMP stimulation of Ca2+ -ATPase activity in rat brain synaptic membranes. AB - The effects of dibutyryl cyclic AMP (db-cAMP) and dibutyryl cyclic GMP (db-cGMP) were tested on Ca2+-ATPase, Mg2+-ATPase, and (Ca2+ + Mg2+)-ATPase activities in lysed synaptosomes prepared from whole rat brains (minus cerebellum). At concentrations from 0.1 to 2.0 mM, db-cGMP produced a selective, concentration dependent increase in Ca2+-ATPase activity. Both db-cGMP and db-cAMP slightly reduced Mg2+-ATPase activity, whereas neither compound had concentration dependent effects on (Ca2+ + Mg2+)-ATPase activity. These findings suggest that the Mg2+-independent, Ca2+-ATPase activity in rat brain is regulated by a cyclic GMP-dependent process. Further, the data provide evidence that the Ca2+-ATPase activity in lysed synaptosomal membranes represents an enzyme that is distinguishable from both the Mg2+ -and (Ca2+ + Mg2+)-ATPase. PMID- 2993520 TI - Muscarinic and nicotinic cholinergic binding sites in the terminal abdominal ganglion of the cricket (Acheta domesticus). AB - Cricket (Acheta domesticus) terminal abdominal ganglia (TG) contain high concentrations (approximately 2 pmol/mg protein) of muscarinic and nicotinic cholinergic binding sites, based on the capacity of TG to bind specifically the labelled ligands L-[3H]quinuclidinyl benzilate ([3H]QNB) and [125I]alpha bungarotoxin ([125I]alpha-BGT) with high affinity. For both ligands, binding is saturable and reversible. Competitive displacement experiments indicate that the [3H]QNB and [125I]alpha-BGT binding sites probably represent pharmacologically distinct classes of putative TG acetylcholine receptors (AChRs). Results from physiological recording and autoradiographic localization experiments demonstrate that a portion of the putative nicotinic AChRs is localized in synaptic regions of the well-characterized cercal sensory-giant interneuron pathway in the TG, where they are likely to serve as functional synaptic AChRs. Unlike nicotinic ligands, muscarinic agents do not appear to be pharmacologically active in this pathway. Therefore, in the insect CNS, putative muscarinic and nicotinic AChRs coexist at high density, but can be pharmacologically distinguished from one another on the basis of criteria derived from both ligand binding and physiological methods. PMID- 2993521 TI - Phosphorylation of the postsynaptic density glycoprotein gp180 by Ca2+/calmodulin dependent protein kinase. AB - Postsynaptic densities (PSDs) were prepared by the aqueous two-phase extraction of synaptic membranes in the presence of n-octyl glucoside. Incubation of postsynaptic densities with [gamma-32P]ATP resulted in the incorporation of 32P into a range of proteins. Isolation of glycoproteins from 32P-labelled PSDs by affinity chromatography on concanavalin A-agarose identified the postsynaptic glycoprotein of apparent Mr 180,000 (gp180) as a substrate for endogenous protein kinase(s). When the phosphorylation reaction was performed in the presence of Ca2+ and calmodulin, there was an overall 13-fold increase in the phosphorylation of PSD proteins. The largest effects of calmodulin were associated with two proteins of molecular weights 51,000 and 60,000, which showed average calmodulin dependent increases in phosphorylation of 68-fold. The phosphorylation of gp180 was increased 7.5-fold in the presence of calmodulin. Fifty percent of maximum phosphorylation of proteins and glycoproteins occurred with a free Ca2+ concentration of 0.3 X 10(-6) M. The amounts 12.6 micrograms/ml and 9.1 micrograms/ml of calmodulin were required for 50% of maximum phosphorylation of proteins and glycoproteins, respectively. Peptide mapping experiments identified three major phosphorylation sites in gp180. The phosphorylation of all three sites was increased in the presence of calmodulin. Phosphoamino acid analysis of gp180 revealed that [32P]phosphoserine and [32P]phosphothreonine were both produced during the phosphorylation reaction, with phosphoserine being the predominant product. The phosphorylation of both amino acids was increased in the presence of calmodulin. [32P]phosphotyrosine was also identified as a product of the phosphorylation of gp180. PMID- 2993522 TI - In vitro phosphorylation of microtubule-associated protein 2: differential effects of cyclic AMP analogues. AB - Microtubules purified from brain tissue contain endogenous cyclic AMP (cAMP) dependent protein kinase activity, and microtubule-associated protein 2 (MAP2) is the major substrate. Beef brain microtubules were prepared and used as a model system to study the differential effects of rationally selected cyclic nucleotide analogues on microtubule receptor protein kinase. Data are presented to indicate that the following molecular interactions are essential for activation of the phosphorylation of MAP2: (a) hydrogen bond formation toward the 2', 3', or 5' position, (b) interaction with phosphorus, and (c) no hydrogen bonds but hydrophobic interactions at the base moiety. Thus, the activation mechanism of the type II protein kinase associated with brain microtubules resembles the mechanism found in protein kinases of other systems. In addition, we have studied the effect of the two diastereomers of adenosine 3',5'-monophosphorothioate (cAMPS). The (Sp)-cAMPS isomer was found to activate MAP2 protein kinase, whereas the (Rp)-cAMPS isomer had no activating effect. In contrast, this compound was able to inhibit cAMP-stimulated MAP2 phosphorylation and thus acts as an antagonist of the Sp diastereomer and cAMP. Hence, this analogue provides a useful means to clarify further the effect of cAMP-dependent phosphorylation on functional properties in microtubules in general. PMID- 2993524 TI - Photoaffinity labelling of benzodiazepine receptors: lack of effect on ligand binding to the nucleoside transport system. AB - This study was undertaken to investigate the possibility of an allosteric interaction between benzodiazepine receptors and the CNS nucleoside transport system. Irreversible (photoaffinity) labelling of the benzodiazepine receptors in guinea pig cortical membranes resulted in a marked reduction in the binding (Bmax) of both [3H]flunitrazepam (71%) and [3H]ethyl-beta-carboline-3-carboxylate (22%) to the benzodiazepine receptors but had no effect on the binding of [3H]nitrobenzylthioinosine to the nucleoside transport system. Furthermore, although photoaffinity labelling resulted in a significant decrease in the affinities of flunitrazepam (approximately equal to 16-fold) and dipyridamole (approximately equal to sevenfold) for the [3H]Ro 15-1788 binding site of the benzodiazepine receptor complex, the affinities of these compounds for the nucleoside transport system were unaltered. These results suggest that the CNS nucleoside transport system and the benzodiazepine receptor complex are distinct, noninteractive ligand recognition sites. PMID- 2993523 TI - Calcium enhances in vitro free radical-induced damage to brain synaptosomes, mitochondria, and cultured spinal cord neurons. AB - Preincubation of rat brain synaptosomes with xanthine and xanthine oxidase (X/XO) in Ca2+-free Krebs buffer resulted in a 27% inhibition of synaptosomal gamma aminobutyric acid (GABA) uptake. Addition of 1.5 mM CaCl2 increased the inhibition with X/XO to 46%, and inhibition was essentially complete when the calcium ionophore A23187 also was included. In other studies, preincubation of purified rat brain mitochondria with the combination of X/XO and 4 microM CaCl2 produced a significant (38%) decrease in state 3 respiration with glutamate/malate as substrate that was not seen with either X/XO or Ca2+ alone. Similar results were obtained using cultured mouse spinal cord neurons in which incubation with X/XO/ADP/FeCl2 and A23187 produced membrane damage as assessed by a 32% reduction of neuronal Na+, K+-ATPase activity. Neither X/XO/ADP/FeCl2 nor A23187 alone caused detectable inhibition. These results demonstrate the synergistic damaging effect of free radicals and Ca2+ on membrane function. In addition, they suggest that free radical-induced peroxidation of membrane lipid, occurring focally during complete or nearly complete ischemia in vivo, could result in intense cellular perturbation when coupled with increased intracellular Ca2+. PMID- 2993525 TI - Effect of iron chelators on dopamine D2 receptors. AB - Nutritional iron deficiency induced in rats causes a selective reduction of [3H]spiperone binding in caudate nucleus. This effect can be reversed by iron supplementation in vivo. The possibility that iron may be involved in the dopamine D2 receptor was investigated by examining the effect of various iron and noniron chelators on the binding of [3H]spiperone in rat caudate nucleus. Iron chelators 1,10-phenanthroline, 2,4,6-tripyridyl-s-triazine, alpha, alpha' dipyridyl, and desferrioxamine mesylate inhibited the binding of [3H]spiperone. The inhibition by 1,10-phenanthroline was noncompetitive and reversible. In the presence of FeCl2 or FeCl3, the inhibitory effect of 1,10-phenanthroline was potentiated. Iron salts or chelators were without effect on the binding of [3H]dihydroalprenolol to beta-adrenoreceptors in caudate nucleus; thus the action of iron chelators on the dopamine D2 receptor tends to be selective. Incubation of caudate nucleus membrane prepared from iron-deficient rats with FeCl2 or FeCl3 did not reverse the diminished binding of [3H]spiperone. The present study indicates that if iron is involved in the physiological regulation of dopamine D2 agonist-antagonist binding sites, it is more complex than hitherto considered. PMID- 2993527 TI - Electrophysiological studies in the post-viral fatigue syndrome. AB - Single fibre electromyography (SFEMG) was studied in 40 patients with the post viral fatigue syndrome. These patients were also assessed clinically, serologically, virologically and immunologically. About 75% of the patients had definitely abnormal SFEMG results. This was regarded as evidence of abnormality in the peripheral part of the motor unit. The muscle fibre was the likely site of involvement. PMID- 2993528 TI - Peripheral polyneuropathy and monoclonal gammopathy of undetermined significance. AB - The prevalence of peripheral polyneuropathy in patients with monoclonal gammopathy is known to be higher than in the general population. A prevalence as high as that in the series of Osby et al, who found clinical and/or electrophysiological evidence compatible with peripheral polyneuropathy in 15 of 21 patients has not been reported before. These results could not be confirmed in a study in which 19 patients with benign monoclonal gammopathy were investigated. In contrast there were only two patients with questionable evidence of peripheral neuropathy: one had lower limb symptoms and signs only, the other had evidence of a subclinical polyneuropathy with some abnormalities of nerve conduction. PMID- 2993526 TI - Reversible abnormalities of the Hering Breuer reflex in acrylamide neuropathy. AB - The sensitivity of the Hering Breuer reflex was compared in anaesthetised rabbits before, during and after the induction of acrylamide neuropathy, and was measured as the tracheal pressure which produced 30 seconds of apnoea. After four weeks of acrylamide (400 mg/kg total dose) there was ataxia and the conduction velocity of hindlimb motor nerves was significantly reduced. At this time there was a marked and reproducible reduction in the sensitivity of the Hering Breuer reflex. The ataxia resolved within a month of stopping acrylamide administration. Three months after the cessation of acrylamide the sensitivity of the Hering Breuer reflex had increased significantly but had not returned to normal. PMID- 2993529 TI - Facial reflex examination for assessment of trigeminal nerve involvement in pituitary fossa tumours. AB - Sixteen patients with pituitary fossa tumours with different intrasellar extension have been studied by facial reflex examination, a neurophysiological test for the trigemino-facial pathway. Impaired transmission along the reflex path was shown in patients with proved encroachments on the flexible walls of the cavernous sinuses, but with no tumour spread to the brain stem and facial nerve. The findings were consistent with a subclinical involvement of the first trigeminal division. Tumour removal resulted in recovery in nerve conduction. It is concluded that facial reflex examination is a valuable test for detecting cavernous sinus involvement in pituitary fossa tumours. PMID- 2993531 TI - Effects of IDPN-induced axonal swellings on conduction in motor nerve fibers. AB - Paranodal demyelination produces a reduction of conduction velocity and conduction block. The relative proportions of these changes appear to vary among different demyelinating disorders. In this study we have examined the effects on conduction of paranodal demyelination produced by giant axonal swellings. The axonal swellings were induced in rats by administration of beta, beta' iminodipropionitrile (IDPN). In this experimental model synchronous axonal swellings occur in the proximal region of virtually every alpha-motorneuron without evidence of segmental demyelination or fiber loss. Conduction across the motor neuron was evaluated by two methods: a monosynaptic reflex pathway and intracellular recording from single motor neurons. Increases in the delay across the central region of the monosynaptic reflex pathway began between 2 and 4 days after toxin administration. Intracellular studies confirmed that the slowing occurred across the proximal regions of the motor axons; more distal regions of the motor axons were unaffected. The substantial reduction in conduction velocity over the swollen segment occurs with only moderate evidence of conduction block, as assayed by a reduction in the H-reflex/M-response amplitude ratio. Parallel morphological studies showed that in the enlarged fibers the myelin terminal loops maintained contact with the axon but were displaced from the paranodal region into the internode. The appearance of this "passive" paranodal demyelination correlated closely with the increase in conduction delay. We suggest that the contact maintained by the displaced myelin terminal loops with the axolemma allows saltatory conduction to continue, and explains the paucity of conduction block in this model despite the prominent conduction slowing. PMID- 2993532 TI - Establishment of a human medulloblastoma cell line and its heterotransplantation into nude mice. AB - A permanent cell line arising from a cerebellar medulloblastoma was established and its growth characteristics were investigated. Although the original tumor inoculum failed to take, the cultured cells were readily tumorigenic in nude mice and gave rise to rapidly growing tumors which could be easily subpassaged. The primary tumor showed evidence of both glial and neuronal differentiation, and retention of neuronal differentiation, albeit minimal, occurred in both the cultured neoplastic cells and the nude mouse tumors. Glial differentiation, on the other hand, could not be demonstrated. G-banding analysis of the chromosomes present in the cell line showed that they were exclusively human. PMID- 2993530 TI - Prognostic significance of serum alpha 1-acid glycoprotein in patients with glioblastoma multiforme: a preliminary communication. AB - The relationship between the levels of serum acute phase reactant proteins (alpha 1-acid glycoprotein, alpha 1-antitrypsin, haptoglobin) was investigated in patients with glioblastoma multiforme in relation to their prognosis. Alpha 1 acid glycoprotein was higher in the patients who died within one year after admission than in those with a longer survival time. It is suggested that serum alpha 1-acid glycoprotein profiles provide prognostic information in patients with glioblastoma multiforme. PMID- 2993534 TI - Low-dose deoxycoformycin in lymphoid malignancy. AB - Deoxycoformycin (dCF), a potent inhibitor of adenosine deaminase (ADA), was explored for its antineoplastic potential in 28 patients with advanced lymphoid malignancy. Both normal and malignant B lymphocytes have low levels of ADA activity, and low doses of dCF profoundly inhibit this enzyme in the peripheral blood of patients with chronic lymphocytic leukemia (CLL). The low doses of dCF administered in this trial (4 mg/m2) were not associated with prohibitive toxicity. Five of 28 patients had an objective response. Four additional patients had clinical improvement. No significant difference in the pretreatment ADA activity existed between responding patients and treatment failures. The demonstration of responses to dCF following failure on standard alkylating agents suggests that dCF may not be cross-resistant with current agents used to treat CLL. Additional studies should be pursued using low-dose dCF in patients with advanced malignancy. PMID- 2993535 TI - High-dose cisplatin in hypertonic saline in refractory ovarian cancer. AB - Nineteen previously treated refractory ovarian cancer patients, including 17 who had received standard-dose cisplatin regimens, were treated in a phase II trial with high-dose cisplatin (40 mg/m2 daily for five days with cycles administered every 28 to 35 days). Objective responses were achieved in 6/19 (32%) patients while eight patients had minor responses or stable disease. The median duration of survival from the start of salvage chemotherapy was 12 months for all patients, and 16 months for responding patients. The dose-limiting toxicity was peripheral neuropathy with 37% of patients having severe paresthesias or ataxia. These results indicate that the dose of cisplatin may be an important factor in improving survival in ovarian cancer patients. PMID- 2993536 TI - Phase II study of aziridinylbenzoquinone (AZQ) in patients with central nervous system malignancies: a Southwest Oncology Group Study. AB - AZQ, an alkylating agent with lipophilic characteristics allowing CNS penetration was studied in patients with primary CNS malignancies refractory to surgical and radiotherapeutic modalities. Responses were evaluated by three criteria: neurologic examination, performance status and CT scan of the brain. Improvement in all three parameters with stable or decreasing doses of decadron was required for a partial response. Thirty-six poor risk (prior chemotherapy) patients with Grades III and IV astrocytomas were treated with 30 mg/m2. Three patients had a partial response (14, 17, 60 weeks duration). Two patients had mixed responses (worsening of one disease parameter with improvement in another), four had stable disease and one patient had improvement in neurologic parameters with a stable CT scan. Twenty-six patients had increasing disease. Fifteen good risk patients (no prior chemotherapy) with recurrent grades III and IV astrocytomas were treated at a dose of 40 mg/m2 intravenously every three weeks. There were no objective responses in this group of patients. Three patients with nonastrocytomas were treated and no responses observed. The drug was well tolerated. Myelosuppression in the form of leukopenia and thrombocytopenia was the major toxicity. Myelosuppression required dose reductions in eight patients and discontinuation of therapy due to repeated treatment delays in two patients. AZQ at doses of 30 and 40 mg/m2 given on an intermittent bolus schedule is inactive in patients with Grades III and IV recurrent astrocytoma. PMID- 2993538 TI - Receptive-field and axonal properties of neurons in the dorsal lateral geniculate nucleus of awake unparalyzed rabbits. AB - The intrinsic stability of the rabbit eye was exploited to enable receptive-field analysis of LGNd neurons and optic tract axons in the awake, unparalyzed state. We found eye position to remain within a range of less than 1.0 degrees for periods of 4-5 min, and in some cases for periods in excess of 10 min. Such stability is comparable to that seen in awake monkeys that have been trained to fixate. Receptive fields of dorsal lateral geniculate nucleus (LGNd) neurons were analyzed, and approximately 84% were concentrically organized. This is a higher value than previously reported in this species. In addition, the receptive-field centers of concentric cells were much smaller than those previously reported (mean diameter = 2.5 degrees). Most remaining neurons in the LGNd were either directionally selective (6.5%) or motion/uniform (6.5%). Concentric cells were classified as sustained or transient based on response duration to standing contrast. In the LGNd the receptive fields of sustained concentric cells were predominantly near the horizontal meridian, within the representation of the visual streak, while the receptive-field positions of transient concentric cells were more prevalent in the upper visual field. In the optic tract the receptive field positions of both sustained and transient cells were more evenly distributed than was seen in the LGNd. Sustained and transient concentric cells in LGNd showed primarily nonlinear spatial summation. The receptive-field properties of LGNd neurons were related to geniculocortical antidromic latency. Most LGNd neurons of all receptive-field classes projected axons to the visual cortex. Thus, any intrinsic interneurons in the rabbit LGNd may have receptive field properties similar to those of some principal neurons. There was significant overlap in the distribution of antidromic latencies in neurons of different receptive-field classes. Concentric sustained neurons, however, did conduct somewhat more slowly than did concentric transient neurons. Nonvisual sensory stimuli that resulted in EEG arousal (hippocampal theta activity) had a profound effect on the response duration of most (28/32) sustained concentric neurons. For these cells, the sustained response to standing contrast began to diminish and sometimes disappeared after 2-15 s. However, arousing stimuli that resulted in hippocampal theta activity reestablished the sustained response. Such arousing stimuli usually had little or no effect on the discharge of the cell in the absence of visual stimuli. Arousing stimuli had no effect on optic tract axons with sustained concentric receptive-field properties.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993533 TI - Neuromonitoring. AB - Neuromonitoring--the continuous or intermittent observation of nervous system functions--has become a field of interdisciplinary interest. Basically there are two major applications of neuromonitoring: in the operating theatre and the neurological or neurosurgical intensive care unit. Evoked potential recording, intracranial pressure measurement, serial EEG recording, cerebral blood flow measurement and ultrasound techniques have all been used as monitoring methods. The application of these techniques for operations, intensive care and the evaluation of brain death will be described. PMID- 2993537 TI - Synaptic potentials in motoneurons during fictive swimming in spinal Xenopus embryos. AB - Embryos spinalized at the 3rd to 6th postotic myotome and immobilized in 10(-4) M tubocurarine can respond to a brief skin stimulus with motor root activity suitable for swimming. Embryos spinalized at the more caudal levels give shorter episodes of fictive swimming. We have previously described the synaptic inputs to motoneurons during fictive swimming in intact embryos (23). In the present paper we look to see if similar synaptic inputs are present in spinal embryos and are therefore spinal in origin. All motoneuron firing during fictive swimming is associated with a tonic depolarization that falls away slowly once firing stops, is increased by hyperpolarizing current, and is reduced by depolarizing current. A slow depolarizing potential evoked by lower levels of skin stimulation has similar properties and rate of fall. In 1-2 mM PDA, an excitatory amino acid antagonist, only a small remnant of the depolarization remains, and motoneuron firing stops. The NMDA antagonist 50 microM APV reduces the depolarization less but also blocks firing. Motoneurons fire one spike per swimming cycle, in phase with nearby motor root discharge. Spikes are preceded by a depolarizing prepotential. This increases with hyperpolarizing current, which can block the spike to reveal an underlying depolarizing potential. In phase with motor root discharge on the opposite side of the body, motoneurons receive a midcycle inhibitory postsynaptic potential, which increases with depolarizing current, decreases with hyperpolarizing current, and is blocked by 10(-6) M strychnine. Strychnine, 5 X 10(-7) M, leads first to broadening of motor root bursts then to loss of the alternating swimming pattern of activity, which is replaced by synchronous bursts on both sides of the body. We conclude that the synaptic inputs to motoneurons during fictive swimming in spinal embryos are very similar in properties and pharmacology to those in intact embryos. These inputs, including the tonic depolarization always associated with motoneuron firing during swimming, must be at least partly spinal in origin. PMID- 2993539 TI - Nonspiking local interneuron in the motor pattern generator for the crayfish swimmeret. AB - We describe a type of nonspiking premotor local interneuron (interneuron IA) in the abdominal nervous system of Pacifasticus leniusculus. All of its branches are restricted to one side of the midline. These interneurons are identifiable and occur as bilateral pairs, one neuron on each side of abdominal ganglia 3, 4, and 5. The membrane potential of interneuron IA oscillated in phase with the swimmeret rhythm, a motor pattern generated in each of these ganglia, because the neuron received postsynaptic potentials in phase with the rhythm. Sustained hyperpolarization of an individual interneuron IA initiated generation of the swimmeret rhythm in all the ganglia of a quiescent nervous system. Sustained depolarization stopped the swimmeret rhythm in all the active ganglia of a nervous system that was generating the rhythm. Currents injected into one interneuron reset the rhythm. Comparisons of the shapes of the IA interneurons in different ganglia showed that they are similar to each other and distinct from other local interneurons in these ganglia. Interneuron IA has a large integrative segment and relatively few branches that are largely restricted to the lateral neuropil, to which all other kinds of swimmeret neurons also project. We conclude that this interneuron occurs only once in each hemiganglion in abdominal segments 3, 4, and 5, and that it is identifiable. Furthermore, this interneuron is an essential component of the circuit in each hemiganglion that generates the swimmeret rhythm. The interneuron was dye coupled to a particular identifiable motor neuron and not to any other neurons. The motor neuron was not dye-coupled to any other local interneurons. The ability of this motor neuron to reset the rhythm is attributed to its being electrically coupled to interneuron IA in its ganglion. PMID- 2993540 TI - Intracellular analysis of the innervation of a crayfish sensory interneuron by regenerating afferents. AB - Mechanoreceptors of the crayfish tail fan have peripheral somata and send their axons to the last (sixth) abdominal ganglion via five bilateral pairs of nerve roots (R1-R5). Comparisons were made between normal crayfish and regenerate preparations in which R4 had been cut and directed back to an extensively denervated sixth abdominal ganglion; 8 to 15 weeks postoperatively, an identified target interneuron (A) in this ganglion was impaled, and its response to water currents, electrical excitation of R4, and stimulation of individual sensory hairs supplying axons to R4 was studied along with several other properties of the pre- and post-synaptic neurons. Normal levels of excitability in A to R4 stimulation were achieved within six weeks as judged by extracellular criteria. Subsequent intracellular analysis revealed that few differences exist between regenerated and normal inputs: probability of (re-) connection, unitary EPSP amplitude and time course distributions, resting membrane potentials, and critical firing levels were comparable in the two groups; input impedance, however, may have been lower in regenerates. Compound electrically elicited EPSPs were similar in amplitude, rise time, and half amplitude width, but differed slightly in latency to onset (regenerates greater than normals). This was accounted for by differences in conduction time to the ganglion in regenerates, and central delay estimates suggest that connections in both groups are monosynaptic. The body root (R1) providing input to A that remained intact in the regenerate preparations increased in efficacy; over 12 postoperative weeks the response of A to R1 activation by water drops steadily increased and at 12 weeks unitary increments to ascending electrical stimulation of R1 were significantly larger than in normals. The response to giant interneuron activation demonstrated that recurrent inhibitory inputs were normal in regenerates. In addition, synaptic depression, normally responsible for behavioral habituation in this system, was comparable across groups. Further, protection from habituation was observed in both normals and regenerates if R4 stimulation was preceded by giant interneuron activation, thus indicating that normal presynaptic inhibitory inputs to the regenerated afferent terminals have also successfully regenerated.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993541 TI - Specificity of early motoneuron growth cone outgrowth in the chick embryo. AB - During development, chick lumbosacral motoneurons have been reported to form precise topographic projections within the limb from the time of initial outgrowth. This observation implies, first, that motoneurons select the appropriate muscle nerve pathway and, second, that they restrict their ramification within the primary uncleaved muscle masses to appropriate regions. Several reports based on electrophysiology and orthograde horseradish peroxidase (HRP) labeling have shown muscle nerve pathway selection to be fairly precise. However, studies based on retrograde labeling with HRP have produced conflicting reports on the extent to which vertebrate motoneurons make projection errors. Since it is difficult to distinguish between true projection errors and HRP leakage when using retrograde labeling, we decided to assess the distribution of labeled growth cones in 25-micron serial plastic sections, following orthograde labeling of identifiable subpopulations of motoneurons during the period of initial axon outgrowth. Examination of a large number of muscle nerves revealed no segmentally inappropriate axons, confirming earlier reports that muscle nerve pathway selection is very accurate. In addition, we observed that growth cones take widely divergent trajectories into the same muscle nerve, suggesting that growth cones are responding independently to some specific environmental cue rather than being passively channeled at this point. The distribution of labeled growth cones within the muscle masses provided direct evidence that motoneurons did not at any time project to obviously inappropriate muscle regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993542 TI - Opiate- and alpha 2-adrenoceptor-induced hyperpolarizations of locus ceruleus neurons in brain slices: reversal by cyclic adenosine 3':5'-monophosphate analogues. AB - The purpose of this study was to investigate the ionic and second messenger mechanisms underlying the hyperpolarizations induced by the selective alpha 2 adrenoceptor agonist clonidine and the opiate agonist morphine in the locus ceruleus. Intracellular recordings were carried out in rat brain slices, and drugs at known concentrations were administered in the perfusate. The cyclic adenosine 3':5'-monophosphate (cAMP) analogues 8-bromo-cAMP and dibutyryl cAMP, while not altering basal activity, reversed the hyperpolarizations induced by clonidine or morphine. In contrast, administration of the parent compound adenosine failed to affect these responses. These results are consistent with previous biochemical studies suggesting that alpha 2-adrenergic and opiate agonists might signal their actions by reducing intracellular cAMP levels. Under manual voltage clamp, both clonidine and morphine elicited outward currents. The algebraic sum of the individual currents elicited by morphine and clonidine significantly exceeded the actual current elicited by their co-administration. This nonadditivity, as well as the observation that cAMP analogues reverse the morphine- and clonidine-induced hyperpolarizations, suggests that these compounds hyperpolarize locus ceruleus neurons through a shared ionic mechanism the activation of which might be signaled by a decrease in intracellular cAMP. PMID- 2993543 TI - Ouabain-evoked norepinephrine release from intact rat sympathetic neurons: evidence for carrier-mediated release. AB - Inhibition of the Na,K-ATPase is known to cause transmitter release from many neurons. The mechanism of [3H]norepinephrine release was examined in primary cultures of sympathetic neurons. Ouabain caused [3H]norepinephrine release at concentrations that produced 80 to 90% inhibition of Na,K-ATPase activity. The effect of ouabain was compared with the effects of high K+ and the Ca2+ ionophore A23187. [3H]Norepinephrine release elicited by depolarization with high extracellular K+ was dependent on extracellular Ca2+, was unaffected by tetrodotoxin, was potentiated by reducing extracellular Na+, and was potentiated by the norepinephrine uptake inhibitor desipramine. These are the results expected if high K+ causes release by exocytosis, and if blockade of the Na+ dependent norepinephrine uptake system increases the net measurable release of the transmitter. The Ca2+ ionophore A23187 caused [3H]norepinephrine release that was not dependent on extracellular Ca2+ but which was like the release elicited by high K+ in other respects. Release elicited by ouabain was independent of extracellular Ca2+, was delayed for several hours by tetrodotoxin, was inhibited by reducing the concentration of extracellular Na+, and was inhibited by desipramine. These results suggest that the measured increase in transmitter release is secondary to a rise in the concentration of intracellular Na+. The data are consistent with the hypothesis that at high levels of pump inhibition, ouabain elicits nonvesicular [3H]norepinephrine release through reversal of the Na+-dependent plasma membrane carrier. PMID- 2993544 TI - The postnatal development of the association projection from visual cortical area 17 to area 18 in the cat. AB - The postnatal development of the association projection from area 17 to area 18 was studied in normal and binocularly deprived kittens between 1 and 28 days of age, using retrograde transport of horseradish peroxidase conjugated with wheat germ agglutinin. The positions of injection sites in the visual cortex, defined in relation to the borders of visual areas 17, 18, and 19 located in Nissl- and cytochrome oxidase-stained sections, were confirmed by observing the patterns of labeling of cells in the lateral geniculate nucleus. The association projection is present and is arranged at least roughly topographically from birth onward; at all ages it arises from cells in both the superficial layers (II, III, and the upper part of IV) and the deep layers (V and VI). In older kittens (20 days or more), however, the origin of the pathway is principally from the upper layers, as in adult cats, whereas in younger animals the projection arises roughly equally from cells in superficial and deep laminae. Initially, the association neurons in area 17 are distributed uniformly along each lamina. Periodic clustering of labeled cells in the upper layers can just be discerned at 10 days, and this patchiness has reached its adult clarity by 20 days, at which stage the projection from the lower layers is greatly diminished. Binocular deprivation until the age of 28 days did not prevent these developmental changes in the projection. Various controls established that the patterns of labeling seen in this study were not due to direct spread of tracer into area 17, to uptake of tracers by fibers-of-passage, or to transcellular transport via the thalamus. PMID- 2993545 TI - Ontogenetic changes in the cyclic adenosine 3',5'-monophosphate-stimulatable phosphorylation of cat visual cortex proteins, particularly of microtubule associated protein 2 (MAP 2): effects of normal and dark rearing and of the exposure to light. AB - Based on a theory that a norepinephrine-stimulated cascade of events resulting in an increase of intracellular cyclic adenosine 3',5'-monophosphate (cAMP) modulates the state of plasticity for the receptive field property of visual cortical neurons, we have followed the ontogenetic changes in cAMP-stimulated phosphorylation of proteins in whole homogenates obtained from developing visual cortices of cats. In vitro phosphorylation was assayed with and without cAMP and the cAMP-dependent protein kinase, and the phosphoproteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were counted for 32P incorporated from [gamma-32P]ATP. It was found that the regulatory subunits of the cAMP-dependent protein kinase are present and fully active by birth, whereas the synapsin content increases at a rate concomitant with synaptogenesis. These ontogenetic developments are not influenced by dark rearing (DR) from birth, a procedure which postpones the onset of the critical period (CP) for plasticity. By contrast, the cAMP-stimulatable phosphorylation of microtubule-associated protein 2 (MAP 2), which under normal rearing conditions increases from birth to the second month, is strongly modulated by the presence of light in the environment. After DR for various periods, kittens were subsequently exposed to light so as to trigger the onset of the CP that had been postponed. A few hours of light were sufficient to cause a large increase in the in vitro phosphorylation of MAP 2. This effect is not observed in the auditory cortex or the lateral geniculate nucleus of the same animals, or in the visual cortex of normally reared cats which were then dark reared in adulthood. But this effect was seen in the visual cortices of cats following 5 months of DR from birth, animals which by chronological age have passed the CP, presumably because the onset of the CP was extended by the DR procedure. The cAMP-dependent phosphorylation of MAP 2 (and its dephosphorylation) may be an important factor for determining the state of plasticity in the CP through its affecting the dendritic cytoskeletal organization involving tubulin and actin. PMID- 2993546 TI - Regeneration of axons and synaptic connections by touch sensory neurons in the leech central nervous system. AB - In studies of axonal regeneration, it has been difficult to determine (a) whether growth along the normal pathway is important for restoration of connections with previous targets and (b) whether the new synapses resemble the old in strength and location. To address these problems at the level of individual nerve cells, we have studied touch (T) sensory neurons in the leech after their axons have been severed and we have confirmed that their axons regenerate electrical connections with some of their usual synaptic targets in the central nervous system. Injections of horseradish peroxidase and Lucifer Yellow dye into separate T cells in unoperated animals showed that T cell axons typically run close to one another within single ganglia or from ganglion to ganglion. Knowledge of one T cell's arborizations thus revealed the groundplan of others in the same ganglia and the sites of apparent contact with its synaptic targets. For regenerating axons, those sprouts that encountered the normal pathway (as marked by homologous axons) grew preferentially along it. Despite the striking coincidence of old and new pathways, regenerated branching patterns within the ganglionic neuropils were usually incomplete and sometimes had atypical branches. Synaptic connections with normal targets (other T cells as well as S and C cells) were abnormally weak physiologically. The numbers of apparent contacts seen with the light microscope were also lower than normal. In addition, the strength of the synaptic potentials, normalized to the number of contacts (calculated as microvolts per contact), was generally smaller in the regenerated connections than in the controls, and smallest at earliest times, during the first 6 weeks following injury. It thus appears to be characteristic of T cell regeneration that axon regrowth is aided by the recognition of specific pathways and that successful regeneration, as assayed anatomically and physiologically, occurs frequently but usually incompletely. PMID- 2993547 TI - Isolation of a clone coding for the alpha-subunit of a mouse acetylcholine receptor. AB - The mouse cell line BC3H-I synthesizes an acetylcholine receptor (AChR) with the pharmacological properties of a muscle nicotinic cholinergic receptor. We have purified mRNA from this cell line and used the size-fractionated poly(A)+RNA to produce a cDNA library of approximately 50,000 clones. The library was screened with a subclone containing genomic sequences coding for the putative acetylcholine-binding site of the alpha-subunit of chicken AChR. We obtained a plasmid, pMAR alpha 15, with a 1,717-base pair insert. The insert cDNA has 26 nucleotides at the 5'-end which code for a portion of the signal peptide followed by a single open reading frame of 1,311 nucleotides which code for a protein of 49,896 daltons. The insert has 377 bases of 3'-untranslated sequence with 3 polyadenylation sites. Radiolabeled plasmid DNA has been used to identify homologous RNA species of about 2 kilobases in Northern blot analyses of poly(A)+ selected RNA from BC3H-I cells. A similar size mRNA is seen in innervated mouse diaphragm and leg muscle, and both mouse and rat brain. Comparisons of the deduced amino acid sequence of the mouse AChR alpha-subunit with Torpedo marmorata, T. californica, chicken, human, and calf sequences show overall homologies of 80%, 80%, 86%, 96%, and 95%, respectively. More detailed analyses reveal a non-random distribution of amino acid substitutions in several structural domains. Based on the absolute conservation of cysteine residues, a new model for the arrangement of the disulfide bonds in the extracellular portion of the alpha-subunit is proposed. PMID- 2993548 TI - Immunosuppressive treatment of multiple sclerosis. AB - Multiple sclerosis is one of the most common causes of chronic neurologic disease in adults. Although the exact pathogenesis of multiple sclerosis is unknown, evidence suggests that it is an autoimmune disease. Recent studies have shown a possible significant beneficial effect of cyclophosphamide in the treatment of multiple sclerosis. This article will discuss the rationale of immunosuppressive therapy and identify nursing measures required in such a clinical trial. PMID- 2993549 TI - Compounds which mediate gallium-67 transfer from lactoferrin to ferritin. AB - The influence of various low molecular weight compounds on the transfer of 67Ga from human lactoferrin (LF) to horse spleen ferritin (HoFE) has been examined in vitro. When LF*67Ga complex was placed in competition with HoFE using a dialysis system the initial transfer rate (TR) of 67Ga to HoFE was slow and continuous. In the presence of 1 mM pyrophosphate (PPi) ascorbate and adenosine triphosphate (ATP), the TR was dramatically enhanced. This effect was concentration sensitive since reduction of the ATP to 0.1 mM eliminated the enhancement. Other intracellular compounds did not significantly influence the TR. Although PPi and ascorbate ions yielded larger TR's, ATP was more effective in the promotion of 67Ga transfer to HoFE. When the LF/HoFE concentration ratio was decreased, in the presence of ATP, the transfer of 67Ga was significantly increased. These results suggest that ferritin present intracellularly could remove and retain 67Ga entering the cell in the form of a LF*67Ga complex. Moreover, increased synthesis of ferritin and cytosolic phosphate compounds would appear to enhance this process. PMID- 2993550 TI - Thallium-201 diethyldithiocarbamate: an alternative to iodine-123 N-isopropyl-p iodoamphetamine. AB - The study of cerebral blood flow by single photon emission computed tomography (SPECT) requires lipophilic radiopharmaceuticals. The high cost and limited availability of N-isopropyl-p-[I-123]-iodoamphetamine ( [123I]IMP) led us to search for alternatives. Following our recent development of thallium-201 diethyldithiocarbamate ( [201TI]DDC), we have compared the brain uptake of [123I]IMP and [201TI]DDC in rabbits. The brain bound 1.14 +/- 0.28% (s.e.m.) of the dose of the injected [123I]IMP and 1.46 +/- 0.28% of the [201TI]DDC. Brain activity of [201TI]DDC remained stable from 1.5 min after injection up to at least 1 hr. The [201TI]DDC uptake was more instantaneous than that of [123I]IMP. The ratios of gray to white matter distribution were about equal: 1.41 for [123I]IMP and 1.44 for [201TI]DDC. The lungs retained 8.32% of the dose of [123I]IMP and only 0.53% of the [201TI]DDC. In brain macroautoradiography [201TI]DDC yielded images of good quality with excellent demarcation of gray and white matter, persisting for at least 45 min after injection. We conclude that [123I]IMP and [201TI]DDC are equally suitable for blood flow study of the rabbit brain. The first human tomographic results obtained in two healthy volunteers demonstrate that clinical application of SPECT [201TI]DDC may be feasible. PMID- 2993551 TI - Factors affecting ventricular volumes determined by a count-based equilibrium method. AB - Using a 99mTc-filled source ("ventricle") in an elliptical torso phantom, we analyzed the effect of source depth, region of interest (ROI) size, background concentration and source shape on volumes determined by an attenuation-corrected count-based equilibrium method. The calculated volume of a 96 cc sphere decreased linearly from 103 to 82 cc with increasing depth from 4 to 18 cm [vol = -1.48 X depth (cm) + 109, r = 0.99]. The calculated volume of the same sphere imaged at a depth of 9 cm increased from 98 to 117 cc with ROI sizes increasing from 161 to 1,369 pixels (1 pixel = 0.17 cm2). With increasing background concentration from 0-2 microCi/ml calculated volumes decreased from 95 to 85 cc (vol = -5.3 X background concentration (microCi/ml) + 95, r = 0.97). However, with correction for over-subtraction of background, increasing background activity caused no decrease in calculated volume (mean = 95 cc, s.d. = 1). Calculated volumes for the sphere and various cylinders were accurate, while those for cones were up to 37% lower for actual volumes ranging from 56-608 cc. This study demonstrates that multiple factors produce variability in count-based determination of phantom volumes. A careful consideration of the interaction of these factors with the edge-detection and computational algorithms is required. PMID- 2993552 TI - Renal handling of technetium-99m DMSA in rats with proximal tubular dysfunction. AB - The renal handling of technetium-99m dimercaptosuccinic acid ([99mTc]DMSA) was studied in rats before and after treatment with Na-maleate (2 mmol/kg i.v.). In the control period, when measured 2 hr after the intravenous injection of [99mTc]DMSA, 39.9% of the injected dose was in the kidneys and 14.6% was in the bladder. After Na-maleate treatment, only 6.4% of the injected dose of [99mTc]DMSA was retained in the kidneys while 37.9% was found in the bladder. Subsequent studies revealed that Na-maleate produced a fall in the glomerular filtration rate, the effective renal plasma flow, and a generalized proximal tubular dysfunction. The latter was characterized by polyuria and an increased excretion of glucose, protein, albumin, calcium, and inorganic phosphate. It was concluded that proximal tubular dysfunction markedly alters the renal handling of [99mTc]DMSA. Whether this augmented urinary excretion is due to an inhibition of reabsorption or an enhanced cellular efflux of [99mTc]DMSA remains to be answered. PMID- 2993553 TI - A practical method of image enhancement by interactive digital filtering. AB - A generalized form of the Wiener digital filter is described in which the user interactively varies two filter parameters to produce any desired number of filtered versions of a given image with differing blends of sharpening and smoothing qualities. Filtering is performed sufficiently rapidly, even without use of an array processor, to make application of the filter truly interactive. Use of the filter is illustrated with typical nuclear medicine images. PMID- 2993554 TI - Theory: basis for the study and practice of nursing education. AB - The theoretical base for nursing education is described as a hierarchy made up of nursing's metaparadigm, conceptual models, and theories. The types of theories needed for the practice of nursing education and the research methods used to develop these theories are discussed. Criteria to be met by a theory before it may be used in nursing education practice are identified. PMID- 2993555 TI - Demographic characteristics of RN and generic students: implications for curriculum. AB - The overwhelming majority of nurses today continue to be prepared at the diploma or associate degree level despite the call of nursing leadership to make the BSN degree the minimum credential for entry into practice. Substantial numbers of registered nurses return to school for a BSN. The purpose of the study was to compare and contrast certain demographic characteristics of RN students with the generic BSN student enrolled in the same program. All junior and senior baccalaureate nursing students enrolled in California colleges and universities which admit RN students to NLN-accredited generic baccalaureate nursing degree programs comprised the study population. There were 3,421 students in the total designated population available for sampling. Of these, 251 actually participated in the study. Findings and implications for Nursing Educators are discussed within the context of the manuscript. PMID- 2993556 TI - Nursing and medical students' attitudes toward the rights of hospitalized patients. AB - A self-administered questionnaire was used to measure the attitudes of Israeli nursing and medical students toward the rights of hospitalized patients in a cross-sectional study of first and fourth year students. Mean attitude scores were high in all student groups showing strong agreement with theoretical rights. Senior nursing students showed significantly higher scores than first-year nursing students. Eighty-eight percent of all students and 100% of senior nursing students considered a blanket consent procedure inadequate for protecting patient's rights. Senior nursing students assigned responsibility for protecting patients' rights to nurses, doctors, hospital administration, and the patient/family with higher frequency than other student groups. PMID- 2993558 TI - Selected pre-admission and academic correlates of success on State Board Examinations. AB - The purpose of this retrospective study was to examine student performance prior to and following admission to a baccalaureate nursing program in comparison with their performance on the individual sections of the State Board Test Pool Examinations (SBE) and an overall assessment of Pass-Fail on the SBE. The following hypothesis guided the study: It is possible to determine which variables are most influential in predicting success on SBEs. Additional research questions addressed the relationship between theoretical and clinical nursing courses and SBE performance, the pertinence of various demographic variables, and differences between students passing and failing SBEs. The sample was composed of the 139 Class of 1980 graduates for whom SBE results were available. Data pertaining to individual course grades, grade point averages, credit hours earned prior to admission, school attended prior to admission, previous academic degree, race, and all SBE scores were obtained. Stepwise multiple regression analysis revealed the junior or senior year cumulative clinical nursing theory GPA had the highest weighting in each section of the SBE and for Pass-Fail SBE. Cumulative clinical nursing theory GPAs were more highly correlated with SBE performance than were cumulative clinical nursing practicum GPAs. The pre-admission liberal arts and cumulative GPAs were also highly correlated with performance. In addition, the number of credit hours earned prior to admission, school attended, and race of the student were found to significant factors. Significant differences between students passing and failing SBEs were found in regard to their level of performance in course work. PMID- 2993557 TI - Improving clinical decision making in nursing. AB - This article describes the need for effective clinical decision making among nurses. The author believes that responses to the need should begin at the educational level, and offers some thoughts to nurse educators for improvement. Exploring the theory base for decision making with students and coordinating teaching-learning strategies with real opportunities in clinical settings are some of the suggestions made. Nurse educators are urged to encourage competency in decision making and to become role models for others. PMID- 2993559 TI - Teaching nursing research by guided design: a pilot study. PMID- 2993560 TI - Individualizing patient care. AB - Patients are people as well as potential problems. Increasing our awareness of patients' lifestyle preferences, as well as developing more productive ways to interact with them, can promote growth along with recovery. The very least it can do is to facilitate working with patients and reduce the negative impact of stress of conflict in a very demanding profession. PMID- 2993561 TI - The improvement of instruction in the clinical area: a challenge waiting to be met. PMID- 2993562 TI - How can a program evaluation be comprehensive and yet cost efficient? PMID- 2993563 TI - Evaluation of a baccalaureate nursing program by alumni and of alumni by their supervisors. PMID- 2993564 TI - Flicker photometry and achromatic-channel structure. AB - It is widely assumed that the near-perfect additivity of heterochromatic flicker photometry implies the existence of an achromatic channel in the visual system, which accurately sums R- and G-cone signals. For flicker photometry to be additive, the channel that detects the flicker stimulus need not add cone signals. PMID- 2993565 TI - Survival of herpes simplex virus on dental handpieces. PMID- 2993566 TI - Surgical treatment of pleomorphic adenoma of the parotid gland: report of 110 cases. AB - One hundred benign and ten malignant pleomorphic adenomas of the parotid gland were analyzed. Follow up ranged from two to 20 years. Of 54 cases of benign pleomorphic adenoma of the parotid gland, recurrence was noted in five cases (9.2%). The five-year recurrence rate was 17.2%. Of the six malignant pleomorphic adenoma, recurrence within the first ten years took place in four patients (66.6%). PMID- 2993567 TI - Lectin probes of glycoconjugates in human salivary gland neoplasms: 2. AB - Lectins are proteins or glycoproteins which exhibit a high affinity for specific sugar molecules. Terminal sugars on cell surface or cytoplasmic oligosaccharides bound to proteins and lipids can be probed with lectins. This study records the lectin-binding characteristics of 20 salivary gland neoplasms and compares them with observations in normal human serous and mucous salivary glands. The results of this study support the current histogenetic concepts for the development of some of the salivary gland neoplasms. PMID- 2993568 TI - [Diagnostic evaluation of RI scintigraphy in parotid gland tumors]. PMID- 2993569 TI - Population and ultrastructural changes in murine alveolar cells following 239PuO2 inhalation. AB - Early changes affecting the principal cellular components of pulmonary alveoli after inhaling 239plutonium dioxide were followed by quantitative and qualitative electron microscopy in mice and rats. Different accumulated doses of a irradiation were achieved. The numbers of alveolar macrophages and interstitial mononuclear cells rose in mice especially after a higher dose of radiation, whilst in rats interstitial fibroblasts were increased. The evidence from mice suggested pronounced secretory activity of type II epithelial cells with subsequent uptake of phospholipid by alveolar macrophages, which developed large cytoplasmic inclusions, but rats were much less severely affected. Pneumonitis was not a feature and with the dosage of radiation employed endothelium escaped structural damage. Sensitivity between species differed, both according to cell type and to intensity of exposure, so demanding caution in the application of experimental findings to man. PMID- 2993570 TI - Sialic acid moieties on surface glycoproteins protect endothelial cells from proteolytic damage. AB - The importance of the glycocalyx in controlling responses to proteinases was studied by enzymic removal of sialic acids from the luminal surface of arterial endothelium. In the perfused rat aorta collagenase induced cell detachment in greater numbers from the desialylated than from the untreated endothelium. Neuraminidase treatment increased by three-fold prostacyclin synthesis by the vessel wall in response to thrombin. The supernatant from activated human neutrophils, containing elastolytic activity, when perfused together with neuraminidase enhanced and prolonged sialic acid release induced by neuraminidase alone. Thus the effects of different proteinases on various endothelial cell functions are potentiated by removal of surface sialic acid residues. PMID- 2993571 TI - Pseudohypoparathyroidism type 1a presenting as congenital hypothyroidism. PMID- 2993572 TI - Discordance for biliary atresia in two sets of monozygotic twins. PMID- 2993573 TI - Vitamin E deficiency is responsible for neurologic abnormalities in cholestatic children. PMID- 2993574 TI - Adenovirus associated with congenital pleural effusion. PMID- 2993575 TI - Isolation of cytomegalovirus from middle ear effusion. PMID- 2993576 TI - Sequelae of acquired cytomegalovirus infection in premature and sick term infants. AB - To assess the risk of long-term sequelae after acquired cytomegalovirus (CMV) infection in premature and sick term infants, 55 CMV infected patients were matched prospectively with 55 control patients and these matched pairs were evaluated at 3 years of age. Sensorineural hearing losses were present in four of 43 CMV infected patients (all mild-moderate) and in two of 43 controls (one severe). The incidence of neurologic sequelae was not increased in CMV infected patients with birth weight greater than 2000 gm. Among patients with birth weight less than 2001 gm, moderately abnormal EEGs were found in four (17%) of 23 CMV infected patients and in one (4%) of 23 controls, and severe handicaps occurred in four (14%) of 29 CMV infected patients and in two (7%) of 29 controls. Severe handicaps in premature infants were significantly (P less than 0.05) associated with early onset of CMV excretion (less than 8 weeks of age) and severe cardiopulmonary disease. Among the premature infants who were documented early excretors, three of 13 had severe neuromuscular impairment, four of 13 had severe handicaps (DQ less than 70, severe neuromuscular impairment, or profound loss of vision or hearing), and an additional four had DQs of 70 to 79. Among their matched control subjects, none of 13 had severe neuromuscular impairment, two of 13 had severe handicaps, and an additional two had DQs between 70 and 79. None of the premature infants who were documented late excretors (greater than or equal to 8 weeks of age) had any neurologic sequelae. The risk of neurologic sequelae and handicap may be increased in premature infants with onset of CMV excretion in the first 2 months of life. PMID- 2993577 TI - The Keyes technique and self-inflicted injuries. Three case reports. AB - Three cases of self-inflicted gingival injuries resulting from the improper use of the Keyes technique are presented. The profession must assume responsibility for studies to determine the safety of the methods of application of the hydrogen peroxide, salt and baking soda mixtures and disseminate this information for the public interest. PMID- 2993578 TI - Evaluation of durapatite ceramic as an alloplastic implant in periodontal osseous defects. II. Twelve month reentry results. AB - Thirteen patients received Durapatite, a hydroxylapatite ceramic (Periograf), as a bone implant material in various types of intrabony defects following internally beveled full thickness flaps, root planing, and defect debridement. All osseous margins and defects were measured from the cemento-enamel junction (CEJ) at specific locations using a standardized periodontal probe. Similarly debrided, nonimplanted defects served as controls. Defect selection as either experimental or control site was based on an alternating defects design after local therapy was completed. Periodontal dressing and systemic tetracycline were used for 10 days. Postsurgical visits for documentation and plaque control were at 10, 20 and 30 days, and 3, 6, 9 and 12 months. Measurements relating to defect changes were made at the 12-month surgical reentry. For evaluation purposes original defect depths were divided into three groups. In Group I (less than 3 mm) defect fill was 1.0 mm (47%) for the implanted defects and 0.3 mm (33%) for the control sites (significantly different at P less than 0.05). In Group II (3-6 mm) defect fill of 1.7 mm (44%) for implanted sites was significantly better (P less than 0.05) than the 0.8 mm (29%) found in control sites. In the deepest group (Group III, greater than 6 mm) Durapatite placement yielded 2.1 mm (32%) of defect fill while debridement alone resulted in 1.8 mm (26%) of fill (P greater than 0.05). Hard tissue responses demonstrated a substantial advantage for use of Durapatite over controls, while soft tissue changes were similar for both.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993579 TI - [Phosphatase activity in porcine brain microtubule proteins]. PMID- 2993580 TI - [Enzymological properties of rat testicular angiotensin I-converting enzyme: comparison with rat pulmonary enzyme]. PMID- 2993581 TI - Influence of inorganic phosphate on the lipid synthesis of a phosphate deregulated mutant of Streptomyces noursei. AB - Phosphate-dependent changes of the mycelial lipid composition were studied in the streptothricin-producing parental strain Streptomyces noursei JA 3890 b/2 and its mutant RG 2. In contrast to its ancestor, the mutant was capable of producing the antibiotic nourseothricin even when large quantities of inorganic phosphate were present in the medium. The apparent insensitivity of the secondary metabolism to phosphate inhibition corresponds to a decreased level of phospholipids in the presence of excessive inorganic phosphate and, during phosphate limitation, to a much higher production of the alkaline phosphatases. A model is discussed which proposed the control by a common genetic element of both the phospholipid and antibiotic production. PMID- 2993582 TI - Bacterial resistance to streptothricins. AB - Resistance to streptothricin was studied in bacteria with different resistance mechanisms. The laboratory-induced streptothricin-resistant mutant E. coli A19 Stcr 2/2/1 showed a high level of cross-resistance to aminoglycosides and other miscoding inducing antibiotics. In contrast, aminoglycosid-resistant E. coli strains with plasmid-determined aminoglycoside modifying enzymes were sensitive to streptothricin. Enzymatic inactivation of streptothricin by acetylation was demonstrated for the streptothricin producing Streptomyces noursei, strain NG13. This strain showed no cross-resistance to miscoding inducing aminoglycosides. PMID- 2993583 TI - [Effect of a cleaning device on teeth. An in vitro study using scanning electron microscopy]. PMID- 2993585 TI - Development of tolerance to chronic barbital treatment in the cerebellar cyclic guanosine monophosphate system and its response to subsequent barbital abstinence. AB - Female Sprague-Dawley rats were placed on a 6-week barbital feeding regimen, previously documented to result in drug tolerance and dependence. Groups of animals were sacrificed on the 2nd and 5th day of each week, up to week 5, or at various time points following drug withdrawal. Cerebellae and cerebral cortices were collected for cyclic GMP (cGMP) measurements. Initial suppressions of cGMP were seen in both the cerebellum and cerebral cortex during early weeks of the feeding regimen. These gradually returned essentially to control levels by the end of 5 weeks, a finding consistent with the development of tolerance to chronic barbiturates in the cGMP system. Barbital was withdrawn after 6 weeks of chronic administration. Four hours after withdrawal, a dramatic elevation of cerebellar cGMP was seen. This elevation was coincident with a significant decline in serum barbital levels and preceded the onset of weight loss, spontaneous seizures and locomotor depression. The elevation of cGMP was less dramatic but still apparent at 72 hr postwithdrawal when all other evaluated parameters had returned to control values. Alterations of cGMP during chronic barbital treatment and subsequent to abrupt barbital withdrawal may reflect aberrations in the function of neurotransmitter pathways involved in regulating cGMP in the cerebellum. Further, the elevation of cGMP after barbiturate withdrawal may itself play a functional role in the manifestation of barbital abstinence. PMID- 2993584 TI - Modulation of beta adrenergic responsiveness by arachidonic acid metabolites in isolated bovine coronary arteries. AB - The present study was designed to determine whether interference with endogenous arachidonic acid metabolism or exogenously administered cyclooxygenase and lipoxygenase products affects the relaxation of bovine coronary artery in response to isoproterenol. Rings of bovine coronary artery were suspended for isometric tension recordings in organ chambers filled with Krebs-Ringer bicarbonate solution (37 degrees C) gassed with 95% O2-5% CO2 (pH 7.4). In depolarized coronary artery rings (35 mM KCI) isoproterenol induced a dose dependent relaxation, which was significantly augmented by the cyclooxygenase inhibitor indomethacin and depressed by arachidonic acid. The mixed lipoxygenase/cyclooxygenase inhibitor phenidone or the lipoxygenase products leukotriene D4 and C4 did not affect beta adrenergic responsiveness. Phenidone antagonized the facilitatory action of indomethacin. Exogenous arachidonic acid in the presence of indomethacin and phenidone depressed the relaxation induced by isoproterenol. Prostacyclin and prostaglandin E2 reduced beta adrenergic responsiveness, which was not affected by indomethacin. The data suggest that arachidonic acid depresses beta adrenergic responsiveness in the bovine coronary artery via cyclooxygenase and some noncyclooxygenase, nonlipoxygenase metabolites. Lipoxygenase products, other than leukotrienes D and C, may have a facilitatory action. PMID- 2993587 TI - Lead induction of crop dysfunction in pigeons through a direct action on neural or smooth muscle components of crop tissue. AB - Lead acetate solutions administered p.o. to pigeons produce crop stasis. Crop dysfunction may be an indirect effect on crop activity by a direct interaction with the cerebellum or some other site associated with lead-induced ataxia. Alternatively, crop stasis may be due to the direct interaction of lead with sites on the smooth muscle or neural elements in crop tissue. A behavioral test for ataxia was performed on pigeons given lead by crop intubation or i.m. injection. Blood lead concentrations were also monitored. Lead-induced ataxia was separable from lead-induced crop dysfunction depending on the route of lead administration, suggesting that lead-induced crop stasis is not secondary to toxicity at a site associated with ataxia. Intramuscular treatment produced crop stasis more readily than did crop intubation. This probably reflects different mechanisms of absorption and metabolism. A Tris-succinate medium was devised which accommodated the solubility characteristics of lead, permitting studies of crop tissue in vitro. Lead chloride added to crop tissue in tris-succinate medium caused a concentration-related reversible relaxation. Crop circular muscle was more sensitive to Pb++ than was longitudinal muscle, in agreement with the effects of other agonists. The EC30 of crop circular smooth muscle in plasma was 1000 microM PbCl2 compared to 3 microM in Tris-succinate medium. The results suggest that lead induces crop dysfunction by acting either directly on crop smooth muscle or on neural elements in crop tissue. PMID- 2993586 TI - Morphine-induced mydriasis and inhibition of pupillary light reflex and fluctuations in the cat. AB - Morphine has species-characteristic effects on pupillary size The effects of morphine on pupillary size, fluctuations and the light reflex were tested with an infrared video pupillometer in the gallamine-paralyzed cat. Compared with saline or base-line responses, i.v. morphine (0.06-1.5 mg/kg) caused a dose-related decrease in the light reflex and fluctuations but increased pupil size. Naloxone (1-100 micrograms/kg i.v.), injected 1 h after morphine, reversed all pupillary effects. Levorphanol (0.5 mg/kg i.v.) had pupillary actions like those of morphine, but dextrophan (0.5 mg/kg i.v.) was inactive. Sympathectomy did not alter the morphine response. It was concluded that morphine disrupts parasympathetic innervation of the iris through interactions with opiate receptors, some of which are in the brain. The morphine-induced changes on the light reflex and fluctuations in the cat are opposite those reported in the rat and rabbit. These results enlarge on the familiar species-dependent effects of opiates on pupillary size. PMID- 2993588 TI - Effects of metaphit, a proposed phencyclidine receptor acylator, on catalepsy in pigeons. AB - Metaphit, a derivative of phencyclidine (PCP), irreversibly binds to PCP sites in rat brain homogenates. PCP-induced catalepsy in pigeons, which is a pharmacologically specific and stereoselective phenomenon, was used to study pharmacological consequences of the proposed covalent bonding of metaphit to PCP sites. Metaphit pretreatment increased the cataleptic effects induced by cumulative doses of PCP-type drugs (i.e., PCP, ketamine and m-amino PCP) and of drugs that have PCP-like actions (i.e., dexoxadrol, LY 154716 and cyclazocine). Metaphit did not affect pentobarbital-induced loss of righting, head-drop and eye closure. Metaphit itself induced a PCP-like catalepsy. Isobolographic analysis of the interactions between metaphit and PCP-like drugs suggested that metaphit potentiated the catalepsy-inducing effects of these drugs. The possibility that metaphit exerts its potentiating effects by inhibition of PCP biotransformation was evaluated by measuring plasma and brain concentrations of PCP after pretreatment with either metaphit or SKF-525A, an inhibitor of the enzyme systems involved in PCP biotransformation. SKF-525A, but not metaphit, increased brain levels of PCP. The results suggest that metaphit acts not as an antagonist of PCP but as a less-potent, long-acting, specific PCP-like agonist. Potentiation by metaphit of the cataleptic effects of chemically diverse drugs with PCP-like actions does not appear to be based on inhibition of the enzyme systems involved in metabolism of those drugs. PMID- 2993589 TI - Ventilatory response to intravenous methionine enkephalin in awake dogs. AB - In conscious, microfilaria-free, adult mongrel dogs, i.v. bolus administration of methionine enkephalin (Met5-ENK) produced a transient elevation of both inspiratory minute ventilation (VI) and heart rate (HR). Both VI and HR increased progressively with increasing doses of Met5-ENK over the range of 6 to 18 micrograms/kg, thereafter plateauing at doses up to 36 micrograms/kg. Maximum changes in VI and HR occurred within 30 to 45 sec after injection, both variables returning to control levels in approximately 2 min. In four out of five dogs, mean inspiratory flow (tidal volume/inspiratory time), and consequently, tidal volume, accounted for this enkephalin-mediated increase in ventilation. In one of the dogs, respiratory rate, rather than tidal volume, increased after Met5-ENK. This change in respiratory rate was due to an increase in "effective timing" of the respiratory cycle, the latter defined as the ratio of inspiratory time to total respiratory time. Despite significant changes in VI and HR, neither end tidal oxygen nor carbon dioxide levels were significantly different from control after i.v. injections of Met5-ENK. Pretreatment with naltrexone methylbromide, a quaternary opiate antagonist that does not cross the blood-brain barrier, abolished all enkephalin-induced changes in VI and HR, thus suggesting that systemic enkephalins modulate ventilation via opiate receptors outside the blood brain barrier. Activation of these receptors produce an increase in both cardiovascular and respiratory activity, as one might expect during stress conditions. These data further support a potential role for peripheral enkephalins as excitatory stress hormones. PMID- 2993591 TI - Action of trimebutine in cat and rabbit colon: evidence of an opioid-like effect. AB - Because trimebutine (TMB) is a drug that modulates colonic motility, we have attempted to determine the modifications of electrical activity underlying the motor effects of this drug. Experiments have been carried out on cat colon in vivo and on rabbit proximal colon in vivo and in vitro. The action of TMB (0.5-2 mg/kg in vivo; 10(-7) M-10(-6) M in vitro) consisted of an initiation or of an increase of spike potential activity that was unaffected by atropine (0.1 mg/kg for the cat, 2-3 mg/kg in vivo and 10(-6) M in vitro for the rabbit) and by hexamethonium (1.5 mg/kg for the cat and 2 mg/kg in vivo or 10(-6) M in vitro for the rabbit). In the cat under atropine, spike activity became cyclical, i.e., bursts of spikes were interrupted by periods of electrical silence. In the cat, TMB decreased the amplitude of excitatory junction potentials. Under atropine, TMB induced rhythmic variations in the amplitude of inhibitory junction potentials for 30 to 40 min. In the rabbit under TMB, excitatory junction potentials were enhanced whereas inhibitory junction potential amplitude was decreased. All the effects of TMB on spontaneous activity as well as on junction potentials are very similar to those observed in a previous work with morphine and enkephalins. The effects of TMB, unaffected by guanethidine, were antagonized by naloxone (2 mg/kg in vivo, 10(-6) M in vitro). These results suggest that opiate receptors, presumably located on intramural neurons, are involved in the effects of TMB.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993590 TI - Neuronal and postjunctional components in the blood pressure effects of dopamine and bromocriptine in rabbits. AB - We have studied the contribution of neuronal and postjunctional dopamine (DA) receptors and of the DA1 and DA2 receptor subtypes to the blood pressure effects of DA and bromocriptine in the rabbit. The norepinephrine release rate, i.e., the rate of entry of endogenous norepinephrine into the plasma, was derived from the plasma level of endogenous norepinephrine and the plasma [3H]norepinephrine clearance. Bromocriptine (40 micrograms kg-1) lowered the norepinephrine release rate and the arterial blood pressure both in anesthetized rabbits and in pithed rabbits with electrically stimulated sympathetic outflow. These effects were antagonized by the selective DA2 antagonist domperidone but not by the selective DA1 antagonist SCH 23390. DA (10-160 micrograms kg-1 min-1) dose-dependently increased the norepinephrine release rate and caused only transient hypotension in anesthetized rabbits. However, after treatment with desipramine, DA did not change the norepinephrine release rate and produced a persistent fall in blood pressure. When DA and domperidone were given simultaneously to desipramine treated rabbits, the hypotensive effect of DA was unchanged, but now DA increased the norepinephrine release rate. When DA and SCH 23390 were given simultaneously to desipramine-treated rabbits, DA failed to lower blood pressure and decreased the norepinephrine release rate. Propranolol did not change the effects of DA in desipramine-treated rabbits. These results suggest that bromocriptine decreases blood pressure by activating ganglionic and/or prejunctional, inhibitory DA2 receptors in the peripheral sympathetic nervous system. DA also activates these receptors, but in addition releases norepinephrine in the manner of an indirectly acting sympathomimetic amine and activates postjunctional vascular DA1 receptors, and the latter seems to be the main component in DA-induced hypotension. PMID- 2993592 TI - Significance of skeletal muscle digitalis receptors for [3H]ouabain distribution in the guinea pig. AB - The importance of specific digitalis glycoside binding sites in skeletal muscle for the digitalis glycoside distribution in the guinea pig was evaluated using [3H]ouabain and [3H]digoxin binding assays. Measurements of [3H]ouabain binding capacity (EOmax) in gastrocnemius and heart muscles in vitro gave values of 474 +/- 15 and 1,092 +/- 39 pmol/g wet wt., respectively, in 4-week-old guinea pigs. Hence the total amount of [3H]ouabain binding sites in skeletal muscle and the heart was around 42,700 and 1,200 pmol, respectively. The apparent dissociation constants (Kd) for ouabain receptor interaction was 0.7 X 10(-7) and 1.5 X 10(-7) M for skeletal muscle and heart, respectively. Comparison of [3H]ouabain and [3H]digoxin binding revealed that these drugs are competitive. From birth to maturity the concentration of [3H]ouabain binding sites in guinea pigs decreased from 803 +/- 58 to 304 +/- 28 pmol/g wet wt. in gastrocnemius muscle and from 1,458 +/- 31 to 1,079 +/- 19 pmol/g wet wt. in the heart. After i.p. injection, measurements of the distribution of [3H]ouabain in plasma, skeletal muscle and the heart showed an almost equal relative specific occupancy of digitalis glycoside receptors in skeletal muscle and the heart: When 10% of the digitalis receptors in the heart were occupied by [3H]ouabain, 13% of those in the skeletal muscles were occupied. It was calculated that 1 hr after the i.p. administration of [3H]ouabain the amount of [3H]ouabain specifically bound to the skeletal muscles and the heart corresponded to 5 times and 1/10 the amount available in the extracellular pool, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993593 TI - Inhibitory effect of acetazolamide on renal tubular reabsorption of NaHCO3 and NaCl in dogs varies inversely with plasma pH. AB - To examine the effect of carbonic anhydrase inhibition on proximal tubular electrolyte reabsorption, plasma pH was altered before and after acetazolamide administration in six volume-expanded dogs during continuous infusion of ethacrynic acid to inhibit transcellular NaCl reabsorption. Plasma pH was altered by changing PCO2, keeping plasma bicarbonate concentration and glomerular filtration rate constant. Linear inverse relationships were obtained between electrolyte reabsorption and plasma pH. Before acetazolamide administration, a change in plasma pH of 0.1 unit from pH 7.4 altered bicarbonate reabsorption by about 10% and sodium and chloride reabsorption remaining during ethacrynic acid infusion by about 6.5%. Administration of acetazolamide (30 mg/kg b.wt.) caused a reduction in electrolyte reabsorption at all plasma pH levels examined. A further reduction occurred after increasing the dose to 100 mg/kg b.wt. The absolute inhibitory effects were almost twice as large during hypercapnia as during hypocapnia whereas the reduction in fractional reabsorption was the same at all plasma pH levels. Both variations in plasma pH and administration of acetazolamide altered the reabsorption of bicarbonate, chloride and sodium in molar ratios of about 1:2:3. Hence, acetazolamide inhibits a constant fraction of the NaHCO3 reabsorption and the associated NaCl reabsorption in the proximal tubules independent of changes in plasma pH. PMID- 2993594 TI - Cardiac effects of adenosine and adenosine analogs in guinea-pig atrial and ventricular preparations: evidence against a role of cyclic AMP and cyclic GMP. AB - The effects of adenosine, the Ri site adenosine receptor agonist (-)-N6 phenylisopropyladenosine (PIA), the Ra site agonist 5'-N ethylcarboxamideadenosine (NECA) and the P site agonist 2',5'-dideoxyadenosine (DIDA) on force of contraction, cyclic AMP (cAMP) and cyclic GMP (cGMP) content and on transmembrane action potential were studied in isolated electrically driven left auricles and papillary muscles from guinea pigs. Furthermore, the effects on adenylate cyclase activity in a particulate membrane preparation were investigated. In the auricles, adenosine, PIA and NECA had negative inotropic effects which were accompanied by a shortening of the action potential. Theophylline antagonized these effects which are likely mediated by R site adenosine receptors. DIDA was ineffective. Except for a small positive inotropic effect of adenosine the analogs were ineffective in the papillary muscles. None of the mechanical effects was accompanied by a change in cAMP and cGMP content in the intact preparations. In the broken cell preparation PIA and NECA had no effect on adenylate cyclase activity. Adenosine and DIDA inhibited the enzyme. The latter effects can be classified as P site-mediated effects. In conclusion, distinct mechanical, i.e., negative inotropic effects of adenosine and its analogs in the heart are observed in auricular preparations only. These effects are unlikely to be related to the cAMP and/or cGMP system. Instead, they are probably due to a direct shortening of the action potential which, in turn, is conceivably due to an increase in K+ outward current and a secondary decrease in Ca++ inward current. This effect is apparently mediated by cardiac R site adenosine receptors which are not detectably coupled to adenylate cyclase. PMID- 2993595 TI - High-affinity cannabinoid binding sites in brain membranes labeled with [3H]-5' trimethylammonium delta 8-tetrahydrocannabinol. AB - The binding of [3H]-5'-trimethylammonium delta 8-tetrahydrocannabinol (THC) [( 3H]TMA) to rat neuronal membranes was studied. TMA is a positively charged analog of delta 8THC modified on the 5' carbon, a portion of the molecule not important for its psychoactivity. Unlabeled TMA inhibits field-stimulated contractions of the guinea-pig ileum (IC50 = 1 microM) in the same presynaptic manner as delta 9THC. [3H]TMA binds saturably and reversibly to brain membranes with high affinity (KD = 89 nM) to apparently one class of site (Hill coefficient, 1.1). Highest binding site density occurs in the brain, but several peripheral organs also display specific binding. Detergent solubilizes the sites without affecting their pharmacological properties. Molecular sieve chromatography reveals a bimodal peak of [3H]TMA binding activity of approximately 60,000 daltons apparent molecular weight. delta 9THC competitively inhibits [3H]TMA binding potently (Ki = 27 nM) and stereoselectively. For some cannabinoids potency in behavioral and physiological tests parallels their affinity for the [3H]TMA binding site. However, several nonpsychotropic cannabinoids are active at the binding site. PMID- 2993596 TI - Dose- and time-dependent biphasic response to morphine on intestinal migrating myoelectric complex. AB - We investigated the biphasic response to morphine infusion on migrating myoelectric complex (MMC) cycling in seven conscious dogs. Morphine infusions at the rate of 20, 50, 100, 200, 500 and 1000 micrograms/kg/hr were started at 0 or 40% of MMC cycle and continued for 3 hr in each experiment. All infusion rates starting at 40% of MMC cycle initiated the first postinfusion MMC cycle prematurely (P less than .05). The mean period of the first postinfusion MMC cycle decreased for higher infusion rates. After the first premature MMC cycle, higher infusion rates (500 and 1000 micrograms/kg/hr) inhibited MMC cycling completely whereas the lower infusion rates inhibited MMC cycling in a dose dependent manner. When infusion was started at 0% of MMC cycle, the first premature MMC cycle occurred consistently only at 20 micrograms/kg/hr. For higher infusion rates, initiation of premature MMC cycle or inhibition of MMC cycling was dose-dependent. Naloxone blocked both the excitatory and the inhibitory effects of morphine on MMC cycling. Morphine infusions also initiated phase III activity in the postprandial state in a dose-dependent manner. Truncal vagotomy and splanchnectomy did not abolish the excitatory or the inhibitory effects of morphine on MMC cycling. We conclude that morphine infusion has a biphasic effect on MMC cycling. The exact nature of the response depends on the dose of morphine and the time in the MMC cycle when infusion is started. Both the excitatory and the inhibitory responses may be mediated through peripheral mu opiate receptors. PMID- 2993598 TI - [The interstitial nucleus of Cajal: its connections and functional considerations]. PMID- 2993597 TI - Forskolin mimics the hydrosmotic action of vasopressin in the urinary bladder of toads Bufo marinus. AB - Net water flow JW was measured across the urinary bladder of toads Bufo marinus and averaged over periods of 1 min by means of a volumetric, automatic technique. The diterpene forskolin, an activator of adenylate cyclase bypassing the hormonal receptor subunit, induced a rapid, reversible, dose-dependent increase in osmotic water permeability, Pf, very similar to that induced by vasopressin. At 1.1 microM, forskolin induced a half-maximal response. At 5 microM forskolin caused a near maximal response and Pf increased from 1.66 +/- 0.15 to 66.6 +/- 2.99 microns s-1. In bladders pre-exposed to 5 microM-forskolin, further significant increases in Pf were obtained by their subsequent exposure to vasopressin, cyclic AMP, theophylline or serosal hypertonicity. The similarity of the forskolin and vasopressin actions was further demonstrated by the finding that substances causing enhancement (quercetin) or inhibition (trifluoperazine, vanadate, silver, cobalt, manganese and Ca2+-free Ringer solution) of the vasopressin response, induced parallel changes in the forskolin response. Three agents, however, induced dissimilar effects on vasopressin and forskolin: high K+ potentiated vasopressin but inhibited forskolin; methohexital and diamide inhibited vasopressin but had no effect on forskolin. The forskolin-induced hydrosmotic response can be viewed as a new criterion for ascertaining the messenger role of cycle AMP in the the hydrosmotic effect of vasopressin. PMID- 2993599 TI - Acquired immune deficiency syndrome. PMID- 2993600 TI - Comparison of four types of diet using clinical, laboratory and psychological studies. AB - Thirty-seven people of different dietary habits-vegans, ovolactovegetarians, whole-food omnivores and average omnivores-were studied using nine-day weighed food intakes, clinical and laboratory assessments, standard psychology questionnaires, measurements of urinary cortisol and catecholamine levels and questionnaires on life-style and health factors. The vegan diet most clearly approximated current thinking on diet, as expressed in the NACNE Report, but was deficient in vitamin D, riboflavin, and vitamin B12. Cholesterol levels were significantly higher in both diet and serum in all groups compared with the vegans. There was no significant difference in social background, money spent on food, exercise, smoking, stress levels or psychological parameters between groups. PMID- 2993601 TI - Non-specific binding of the fluorescent beta-adrenergic receptor probe alprenolol NBD. AB - The fluorescent beta-adrenergic receptor probe alprenolol-NBD was found to exhibit a high affinity (Kd 3.2 nM) and a low capacity (10 fmol/mg protein) for the beta 2-adrenergic receptor on living Chang liver cells but also a high affinity (Kd 320 nM) for non-beta-adrenergic receptor binding sites with a very high capacity (28,000 fmol/mg protein). Calculations are presented which make clear that less than 3% of the binding of alprenolol-NBD during visualization experiments is beta-adrenergic receptor related. Furthermore, it is shown that besides the downregulation of beta-adrenergic receptors during incubation with isoproterenol, the high-affinity non-beta-receptor binding sites are also deminishing during incubation with isoproterenol. Based on our findings it is concluded that the results of Henis et al. who claimed the visualization of the beta-adrenergic receptor population on Chang liver cells by alprenolol-NBD must be interpreted as an almost completely non-specific fluorescence. PMID- 2993602 TI - Specific increase in renal alpha 1-adrenergic receptors following unilateral renal denervation. AB - The effect of unilateral surgical renal denervation on adrenergic receptors in rat kidney was investigated. Denervation was performed by mechanically stripping the nerves from the renal artery and then painting with phenol. This procedure resulted in a decrease in norepinephrine content to less than 10% of the contralateral kidney. The concentrations of alpha 1-, alpha 2-, beta 1- and beta 2-adrenergic receptors were measured in both kidneys of rats 1 week following left renal denervation. Alpha 1-receptor concentrations increased in the denervated left kidneys relative to the intact right kidneys. Alpha 2-, beta 1- and beta 2-receptor concentrations were not different between left and right kidneys. These results indicate that the different classes of adrenergic receptor in the kidney are differentially regulated by agonist stimulation. PMID- 2993603 TI - Activation of the 1,25-dihydroxyvitamin D3 receptor in cultured rat osteogenic sarcoma cells. AB - Activation of the 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] receptor-hormone complex was studied in-vitro using cytosolic preparations of rat osteogenic sarcoma cell ROS 17/2-8 and a DNA-cellulose assay. We found that salt was required for extraction of the unoccupied receptor indicating its possible nuclear localization. The 1,25(OH)2D3 receptor underwent an activation process similar to other steroid hormones which could be stimulated by heat and salt. At physiological ionic strength 100% of the complexes were, however, activated at 2 degrees C, indicating that the activation process is not absolutely temperature dependent. In contrast to other steroid hormones, 30-50% of the complexes were in an activated state in the absence of heat and salt moreover, alkaline phosphatase and ammonium sulphate had no effect on activation. Activation was also stimulated by ATP and ATP plus 8BrcAMP indicating the possible role of phosphorylation in the activation process; however, further work is required to clarify this point. PMID- 2993604 TI - Dihydropyridine binding sites on transverse tubules isolated from triads of rabbit skeletal muscle. AB - Receptors for the dihydropyridine class of Ca2+ channel antagonists are present on transverse tubules isolated by mechanical disruption of skeletal muscle triads. These observations account for the previously reported presence of nitrendipine binding sites in heavy sarcoplasmic reticulum. Nitrendipine receptors were not found in the terminal cisternae after disruption of the triad junctions. The number of sites in junctional T-tubules (27 pmol/mg) is only half that reported for transverse tubules isolated as free vesicles (pmol/mg). The presence of nitrendipine receptors in that region of the transverse tubules held in close apposition to the SR cisternae is consistent with the architectural requirements for the Ca2+ induced Ca2+ release phenomenon to be the mechanism of excitation-contraction coupling. PMID- 2993605 TI - Gonadotrophin-stimulated cyclic AMP production by granulosa cells from Booroola x Romney ewes with and without a fecundity gene. AB - The influence of follicular size and health on FSH and LH stimulation of cAMP production by granulosa cells in vitro was studied in cells from Booroola X Romney ewes, with (F+) and without (++) a fecundity gene. The granulosa cells were obtained 0-48 h after the initiation of luteolysis on Day 10 of the oestrous cycle by cloprostenol. The highest mean amounts of cAMP produced by granulosa cells challenged with FSH or LH were not significantly different between the genotypes. However, they were achieved using granulosa cells from follicles greater than 3-4 mm in diameter in F+ ewes but from follicles greater than 4 mm in diameter in ++ ewes. Follicles may thus attain ovulatory maturity at a smaller diameter in F+ ewes than in ++ ewes. Granulosa cells from most atretic follicles gave a poor cAMP response to FSH or LH, compared to cells from non-atretic follicles. Granulosa cell responsiveness to FSH was independent of the time the cells were recovered after cloprostenol treatment in F+ ewes, but not in ++ ewes. Cellular responsiveness to LH was independent of time for sheep of both genotypes. There was a significant positive relationship for sheep of both genotypes between the level of aromatase activity in granulosa cells and cellular responsiveness to FSH and LH. PMID- 2993606 TI - FSH influences follicle viability, oestradiol biosynthesis and ovulation rate in Romney ewes. AB - Injection of steroid-free bovine follicular fluid (bFF; 2 X 5 ml s.c. 12 h apart) into anoestrous ewes lowered plasma FSH concentrations by 70% and after 24 h had significantly (P less than 0.01) reduced the number of non-atretic follicles (greater than or equal to 1 mm diam.) without influencing the total number of follicles (greater than 1 mm diam.) compared to untreated controls. Hourly injections of FSH (10 micrograms i.v. NIH-FSH-S12) for 24 h did not influence the number of non-atretic follicles but did negate the inhibitory effects of bFF on follicular viability. Hourly injections of FSH (50 micrograms i.v., NIH-FSH-S12) + bFF treatment for 24 h significantly increased the total number of non-atretic follicles, and particularly the number of medium to large non-atretic follicles (greater than 3 mm diam.) compared to the untreated controls (both P less than 0.01). The 10 micrograms FSH regimen (without bFF) significantly increased aromatase activity in granulosa cells from large (greater than or equal to 5 mm diam.; P less than 0.01) but not medium (3-4.5 mm diam.) or small (1-2.5 mm diam.) follicles compared to controls. The 10 micrograms FSH + bFF regimen had no effect on granulosa-cell aromatase activity compared to the controls. However, the 50 micrograms FSH plus bFF regimen increased the aromatase activity of granulosa cells from large, medium and small non-atretic follicles 2.6-, 8.3- and greater than or equal to 11-fold respectively compared to that in the control cells. Ewes (N = 11) that ovulated 2 follicles had significantly higher plasma FSH concentrations from 48 to 24 h and 24 to 0 h before the onset of a cloprostenol-induced follicular phase (both P less than 0.01) than in the ewes (N = 12) that subsequently ovulated one follicle. Hourly FSH treatment (1.6 micrograms i.v., NIAMDD-FSH-S15) for 24 h but not for any 6 h intervals between 48 and 24 h or 24 and 0 h before a cloprostenol-induced luteolysis also resulted in significant increases (P less than 0.05) in the number of ewes with 2 ovulations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993607 TI - Antidotes for gold (sodium bis[thiosulfato]gold[I]) intoxication in mice. AB - Eight compounds were examined to determine their relative efficacy as antidotes in acute gold intoxication in mice after the intraperitoneal injection of 200 mg/kg of Na3 [Au(S2O3)2] X 2H2O. Of the compounds examined, 2,3 dimercaptosuccinic acid was the most effective antidote. It was noted that with D penicillamine those animals which did not survive died sooner (in a matter of hours) than the corresponding control animals which had received no antidote (about 3 days). In subsequent experiments in which Na3 [Au(S2O3)2] X 2H2O was administered at the lower level of 140 mg/kg, 2,3 dimercaptosuccinic acid showed itself to be capable of reducing kidney gold levels by a factor of about 5 and liver gold levels by a factor of approximately 2. PMID- 2993608 TI - Computer-assisted drug design. PMID- 2993610 TI - Synthesis and pharmacological characterization of 5-(2-dodecylphenyl)-4,6 dithianonanedioic acid and 5-[2-(8-phenyloctyl)phenyl]-4,6-dithianonanedioic acid: prototypes of a novel class of leukotriene antagonists. PMID- 2993609 TI - Stereostructure-activity relationship of opioid agonist and antagonist bivalent ligands. Evidence for bridging between vicinal opioid receptors. PMID- 2993611 TI - Syntheses and differentiating action of vitamin D endoperoxides. Singlet oxygen adducts of vitamin D derivatives in human myeloid leukemia cells (HL-60). AB - Singlet oxygen adducts of various vitamin D derivatives, 6,19-dihydro-6,19 epidioxyvitamin D (vitamin D endoperoxides, 2 and 2'), were chemically synthesized, and their biological activity in inducing differentiation of a human myeloid leukemia cell line (HL-60 cells) was examined. The potency of the endoperoxides derived from vitamin D derivatives possessing the 1 alpha-hydroxyl group such as 1 alpha, 25-dihydroxyvitamin D3 endoperoxides (2b and 2b') was markedly (10(-2)) diminished relative to the respective parent vitamin D compounds. In contrast, 25-hydroxyvitamin D3 endoperoxides [25-(OH)D3 endoperoxides, 2a and 2a'] and their analogues fluorinated at the 24- or 26- and 27-positions were 2.5-10 times more potent than 25-hydroxyvitamin D3 (1a) in spite of the absence of the conjugated triene structure typical of vitamin D compounds. The potency of these vitamin D endoperoxides (2 and 2'), especially those lacking the 1 alpha-hydroxyl group, in inducing differentiation of HL-60 cells was not correlated with their activity in binding to the cytosol receptor for 1 alpha, 25-dihydroxyvitamin D3 (1b). The binding efficiency to the receptor was relatively lower than the differentiating activity. To examine the action of vitamin D endoperoxides, carbon analogues of 25-(OH)D3 endoperoxides, two C-6 epimers of 25-hydroxy-6,19-dihydro-6,19-ethanovitamin D3 (6 and 6'), were synthesized. The carbon analogues (6 and 6') had no potential to induce differentiation of HL-60 cells. These results suggest that vitamin D endoperoxides (2 and 2') express their biological activity probably after being converted to some other compounds. PMID- 2993612 TI - Mechanism of the differentiating action of 25-hydroxyvitamin D3 endoperoxides in human myeloid leukemia cells (HL-60). AB - The action of 25-hydroxy-6,19-dihydro-6,19-epidioxyvitamin D3 [25-(OH)D3 endoperoxides, 2a and 3a] in inducing differentiation of human myeloid leukemia cells (HL-60) was studied by using their radioactive derivatives (2a' and 3a'). When HL-60 cells were incubated with the labeled endoperoxides (2a' and 3a') in serum-free RPMI 1640 medium, no radioactivity was incorporated into either the cytosol or the chromatin fraction of the cells. When the radioactive endoperoxide (2a') was incubated in the culture medium for 3 days, with or without HL-60 cells, about 45% of the compound was similarly converted to 19,25-dihydroxy-6,19 dihydro-6,19-epoxyvitamin D3 (4a) and about 10% to 25-hydroxy-6,19-epoxyvitamin D3 (6a). These two new vitamin D derivatives were synthesized chemically and tested for their biological activities. Both compounds (4a and 6a) were about 2 times as active as 25-(OH)D3 endoperoxides (2a and 3a) and about 7 times as active as 25-hydroxyvitamin D3 (1a) in inducing differentiation of HL-60 cells. The differentiating activity of these compounds was well correlated with their activity in binding to the cytosol receptor for 1 alpha, 25-dihydroxyvitamin D3 in HL-60 cells. The in vitro bone-resorbing activity of 25-hydroxy-6,19 epoxyvitamin D3 (6a) and 25-(OH)D3 endoperoxide (2a) was higher than that of 25 hydroxyvitamin D3 (1a), indicating that the differentiating activity also paralleled the bone-resorbing activity in these vitamin D derivatives. These results suggest that 25-(OH)D3 endoperoxides (2a and 3a) induce differentiation of HL-60 cells and bone resorption after being converted to these two compounds. PMID- 2993614 TI - Synthesis and biological evaluation of phosphonamidate peptide inhibitors of enkephalinase and angiotensin-converting enzyme. AB - The effectiveness of phosphonamidate peptide analogues as inhibitors of rat kidney or human brain metalloendopeptidase (enkephalinase, E.C. 3.4.24.11) and angiotensin-converting enzyme (ACE, 3.4.14.1) has been explored with a series of enkephalin analogues in which the scissile Gly3-Phe4 amide bond has been replaced with a phosphonamidate moiety. These compounds exhibited good inhibitory potency against enkephalinase with several of the analogues having Ki values in the submicromolar range as contrasted to micromolar or higher toward ACE. Within a series of [(N-acylamino)methyl] phosphonamidates there was a dramatic decrease in inhibitory activity against enkephalinase as the N-acyl moiety was substituted with larger, more hydrophobic acyl groups. Likewise, the inhibitory activity of the [(N-acylamino)methyl] phosphonamidates against ACE was attenuated by larger phenylalkyl acyl functionalities, although not to the same degree as against enkephalinase. However, phosphonamidate pentapeptide analogues of (Leu)enkephalin and (D-Ala2,D-Leu5)enkephalin showed good inhibitory potency against both enzymes. Interestingly, these two (Leu)enkephalin phosphonamidate analogues were completely inactive in the electrically stimulated guinea pig ileum and mouse vas deferens preparations. Conformational factors that may be involved in this inactivity are discussed. PMID- 2993615 TI - Synthesis and antiherpes simplex virus activity of 9-[(1,3-dihydroxy-2 propylthio)methyl]guanine. AB - The synthesis of the thio analogue (thio-DHPG, 2) of 9-[(1,3-dihydroxy-2 propoxy)methyl]guanine (DHPG, 1) is described. The synthesis of 2 proceeded via the condensation of acetoxymethyl sulfide 9 with diacetylguanine 10 to give the protected nucleoside analogue 11. Although catalytic hydrogenolysis failed, the benzyl ether functionalities of 11 were successfully cleaved by an acetolysis reaction to furnish 14. Ammonolysis of 14 gave 2, which was also transformed to sulfoxide 15 and sulfone 16. Preliminary in vitro screening indicated that 2 exhibited comparable activity to DHPG against herpes simplex virus type 1 (HSV-1) but was less active against the type 2 virus (HSV-2) and human cytomegalovirus (HCMV). In a mouse encephalitis model (HSV-2), subcutaneous treatment with 2 led to a 53% reduction in mortality at a dose of 100 mg/kg per day. PMID- 2993613 TI - Synthesis and biological activity of 5-thiobredinin and certain related 5 substituted imidazole-4-carboxamide ribonucleosides. AB - A number of 5-substituted imidazole-4-carboxamide ribonucleosides were prepared and tested for their biological activity. Treatment of 5-chloro-1-beta-D ribofuranosylimidazole-4-carboxamide (2) with methanethiol provided 5 (methylthio)-1-beta-D-ribofuranosylimidazole-4-carboxamide (3a). Similar treatment of 2 with ethanethiol or benzenemethanethiol gave the corresponding 5 ethylthio and 5-benzylthio derivatives 3b and 3c. Oxidation of 3a and 3b with m chloroperoxybenzoic acid furnished the corresponding sulfonyl derivatives 4a and 4b. Reductive cleavage of 3c with sodium naphthalene or Na/NH3 gave 5-mercapto-1 beta-D-ribofuranosylimidazole-4-carboxamide (5-thiobredinin, 5). Direct treatment of 2 with sodium hydrosulfide provided an alternate route to 5, the structure of which was established by single-crystal X-ray analysis. 5-Thiobredinin has a zwitterionic structure similar to that of bredinin. Glycosylation of persilylated ethyl 5(4)-methylimidazole-4(5)-carboxylate (6) with 1-O-acetyl-2,3,5-tri-O benzoyl-D-ribofuranose in the presence of SnCl4 provided a quantitative yield of the corresponding tri-O-benzoyl nucleoside 7. Debenzoylation of 7 with MeOH/NH3 at ambient temperature gave ethyl 5-methyl-1-beta-D-ribofuranosylimidazole-4 carboxylate (8). Further ammonolysis of 8 or 7 at elevated temperature and pressure gave 5-methyl-1-beta-D-ribofuranosylimidazole-4-carboxamide (9). All of these ribonucleosides were tested in Vero cell cultures and in mice against certain viruses. Compounds 3a and 3c exhibited significant activity against vaccinia virus in vitro, whereas 4a was effective against Rift Valley fever virus in mice. 5-Thiobredinin failed to exhibit appreciable antiviral or cytostatic activity (against L1210 and P388) in cell culture. PMID- 2993616 TI - N-substituted imidazolines and ethylenediamines and their action on alpha- and beta-adrenergic receptors. AB - A series of N-substituted imidazolines and ethylenediamines were synthesized and examined for their activity in alpha- and beta-adrenergic systems. The length of the intermediate side chain between the catechol and imidazoline ring or the amine of the ethylenediamine segment was shown to affect the adrenergic activity. N-[2-(3,4-Dihydroxyphenyl)ethyl]imidazoline hydrochloride (2) and N-[2-(3,4 dihydroxyphenyl)ethyl]ethylenediamine dihydrochloride (4), both with two methylene groups between the catechol and amine segment, were found to be somewhat selective for alpha 2-adrenergic receptors while 1-(3,4 dihydroxybenzyl)imidazoline hydrochloride (1) and N-2-(3,4 dihydroxybenzyl)ethylenediamine dihydrochloride (3), both with one methylene group between the catechol and amine segment, were more selective for alpha1 adrenergic receptors in a pithed rat model. Of the four compounds examined, only compound 2 showed significant direct activity on beta1- and beta2-adrenergic receptors. PMID- 2993617 TI - Synthesis of new bis(aminoethanethiol) (BAT) derivatives: possible ligands for 99mTc brain imaging agents. AB - In developing new brain perfusion imaging agents for single photon emission computed tomography (SPECT), five 99mTc-labeled neutral bis(aminoethanethiol) (BAT) derivatives capable of crossing the blood-brain barrier are reported. The five ligands are prepared by two versatile synthetic methods that can specifically introduce substituents on one of the carbons between two nitrogens. These ligands formed stable and neutral complexes with the reduced 99mTc, using either Sn(II) or sodium borohydride to reduce sodium [99mTc]pertechnetate. The biodistribution in rats was evaluated with [125I]iodoantipyrine, a freely diffusible tracer, as the internal reference. Compounds with a free hydroxyl group, 6 and 15, showed lower brain uptake. High initial brain uptake was observed for compounds 10 and 14, 1.28 and 2.30% dose/organ, respectively. Compounds of this type may be used as a basis for future structural modification to improve brain uptake and retention. PMID- 2993618 TI - Synthesis and biological activity of modified peptide inhibitors of angiotensin converting enzyme. AB - A series of non-sulfhydryl modified dipeptides related to CI-906, CI-907, and enalapril was prepared in which various isosteric moieties (O, S, SO, SO2) have been substituted for the amino group and in which the proline residue has been replaced with various hydrophobic amino acids. The compounds were evaluated in vitro for inhibition of angiotensin-converting enzyme and in vivo for antihypertensive activity. Compound 7c, the most potent member of this series, had an in vitro IC50 of 1.4 X 10(-8) M and showed modest oral antihypertensive activity at 30 mg/kg in conscious, two kidney, one clip Goldblatt hypertensive rats. Structure-activity relationships are discussed. PMID- 2993619 TI - Ring-substituted 1,1,2,2-tetraalkylated 1,2-bis(hydroxyphenyl)ethanes. 4. Synthesis, estrogen receptor binding affinity, and evaluation of antiestrogenic and mammary tumor inhibiting activity of symmetrically disubstituted 1,1,2,2 tetramethyl-1,2-bis(hydroxyphenyl)ethanes. AB - The syntheses of symmetrically 2,2'-disubstituted derivatives of 1,1,2,2 tetramethyl-1,2-bis(4-hydroxyphenyl)ethane (1) and of 5,5'-, and 6,6' disubstituted derivatives of 1,1,2,2-tetramethyl-1,2-bis(3-hydroxyphenyl)ethane (6) are described (1 and 6 are strong antiestrogens with mammary tumor inhibiting activity exhibiting only slight estrogenic properties): (2,2'-substituents) F (2), Cl (3), OCH3 (4), CH3 (5); (5,5'-substituents) Cl (7); (6,6'-substituents) F (8), Cl (9), OCH3 (10), CH3 (11). The synthesis of 1-11 was accomplished by reductive coupling of the corresponding 2-phenyl-2-propanols with TiCl3 and LiAlH4. The binding affinity of the compounds to the calf uterine estrogen receptor was measured relative to that of [3H]estradiol by a competitive binding assay. With the exception of 7 and 10 all other compounds showed relative binding affinity (RBA) values between 0.5 and 6.4% that of estradiol, 2 (RBA value 6.4), and 8 and 9 (4.0 and 3.5), exceeding those of the corresponding unsubstituted 1 and 6 (3.6 and 3.0). Compounds exhibiting RBA values of greater than 2.5% were evaluated in the mouse uterine weight test. The substituted derivatives showed an increase in uterotrophic and a decrease in antiuterotrophic activity compared to 1 and 6. Compound 2 showed a strong, dose-dependent inhibition on the DMBA induced hormone-dependent mammary tumor of the SD-rat, exceeding that of the parent compound 1. At a dose of 5 mg/kg per day, 2 reduced total tumor area by 47% and caused a complete remission in 74% of the tumors. PMID- 2993620 TI - Benzodiazepine receptor binding and anticonflict activity in a series of 3,6 disubstituted pyridazino[4,3-c]isoquinolines devoid of anticonvulsant properties. AB - A series of 3,6-disubstituted pyridazino[4,3-c]isoquinolines were synthesized and tested for their ability to inhibit the binding of [3H]diazepam to rat brain receptors in vitro. Compounds bearing a phenyl, 4-methoxyphenyl, or methyl group at position 3 and a dialkylamino group at position 6 showed the highest affinity in the binding assay and were subsequently evaluated for their anticonflict and anticonvulsant effects. All of these compounds (5a-1 and 5q) were active in the Vogel rat conflict procedure, but none prevented convulsions in mice induced either by metrazol or bicuculline. 3-Phenyl-6-pyrrolidinylpyridazino[4,3 c]isoquinoline (5d) with a Ki = 11.4 nM in the binding assay exhibited the best potency in the anticonflict assay (MED 5 mg/kg ip) and did not produce neuromuscular impairment at the highest dose tested (50 mg/kg ip). PMID- 2993621 TI - Quantitative evaluation of the beta 2-adrenoceptor affinity of phenoxypropanolamines and phenylethanolamines. AB - The influence of the aromatic moiety of beta-adrenoceptor ligands on the affinity for the beta 2-adrenoceptor has been studied. Three classes of ligands have been examined, viz. N-isopropyl- and N-tert-butylphenylethanolamines and N isopropylphenoxypropanolamines. Computer-assisted analysis of the inhibition by any of these ligands of the specific (-)-[3H]dihydroalprenolol binding to the beta 2-adrenoceptors of a bovine skeletal muscle preparation in the presence of GppNHp (10(-4) M) yielded the affinities of these ligands at pH 7.5. The obtained values were adjusted for the amounts of cations present at this pH value. A significant correlation was found between the calculated lipophilicities and the experimentally determined affinities in the three classes. Furthermore, steric factors seem to play an important role, as these correlations were improved by the introduction of steric parameters for the aromatic substituents in the regression analyses. From the established equations it is concluded that the phenoxypropanolamine derivatives bind to the beta 2-adrenoceptor in a way different from that of the ligands in both ethanolamine classes. PMID- 2993623 TI - Functionalized congeners of adenosine: preparation of analogues with high affinity for A1-adenosine receptors. AB - A series of functionalized congeners of adenosine based on N6-phenyladenosine, a potent A1-adenosine receptor against, was synthesized. Derivatives of the various congeners should be useful as receptor and histochemical probes and for the preparation of radioligands and affinity columns or as targeted drugs. N6-[4 (Carboxymethyl)phenyl]adenosine served as the starting point for synthesis of the methyl ester, the methyl amide, the ethyl glycinate, and various substituted anilides. One of the latter, N6-[4-[[[4 (carbomethoxymethyl)anilino]carbonyl]methyl]phenyl] adenosine, served as the starting point for the synthesis of another series of congeners including the methyl amide, the hydrazide, and the aminoethyl amide. The terminal amino function of the last congener was acylated to provide further analogues. The various congeners were potent competitive antagonists of binding of N6 [3H]cyclohexyladenosine to A1-adenosine receptors in rat cerebral cortical membranes. The affinity of the congener for the A1 receptor was highly dependent on the nature of the spacer group and the terminal moiety with Ki values ranging 1-100 nM. A biotinylated analogue had a Ki value of 11 nM. A conjugate derived from the Bolton-Hunter reagent had a Ki value of 4.5 nM. The most potent congener contained a terminal [(aminoethyl)amino]carbonyl function and had a Ki value of less than 1 nM. PMID- 2993622 TI - Functionalized congeners of 1,3-dialkylxanthines: preparation of analogues with high affinity for adenosine receptors. AB - A series of functionalized congeners of 1,3-dialkylxanthines has been prepared as adenosine receptor antagonists. On the basis of the high potency of 8-(p hydroxyphenyl)-1,3-dialkylxanthines, the parent compounds were 8-[4 [(carboxymethyl)oxy]phenyl] derivatives of theophylline and 1,3-dipropylxanthine. A series of analogues including esters of ethanol and N-hydroxysuccinimide, amides, a hydrazide, an acylurea, and anilides were prepared. The potency in blocking A1-adenosine receptors (inhibition of binding of N6 [3H]cyclohexyladenosine to brain membranes) and A2-adenosine receptors (inhibition of 2-chloroadenosine-elicited accumulations of cyclic AMP in brain slices) was markedly affected by structural changes distal to the primary pharmacophore (8-phenyl-1,3-dialkylxanthine). Potencies in the dipropyl series at the A1 receptor ranged from Ki values of 1.2 nM for a congener with a terminal amidoethyleneamine moiety to a Ki value of 58 nM for the parent carboxylic acid to a Ki of 96 nM for the bulky ureido congener. Certain congeners were up to 145 fold more active at A1 receptors than at A2 receptors. Various derivatives of the congeners should be useful as receptor probes and for radioiodination, avidin binding, and preparation of affinity columns. PMID- 2993624 TI - Replacement of the ribofuranose oxygen of 2-5A derivatives by methylene: synthesis of an aristeromycin analogue of 2-5A core 5'-monophosphate (5'-O phosphoryladenylyl)(2'----5')adenyl(2'----5')adenosine. AB - The ribofuranose oxygens of the three adenosine residues of the 5'-monophosphate of the 2-5A core [adenylyl(2'----5')adenylyl(2'----5')adenosine] were replaced by methylenes through the synthesis of an aristeromycin [9-[(1R,2S,3R,4R)-2,3 dihydroxy-4-(hydroxymethyl)cyclopentyl]adenosine] analogue. In the synthetic approach, the chlorophosphite triester procedure was employed together with the use of dimethoxytrityl and tert-butyldimethylsilyl protecting groups. The final product 14 was bound to the 2-5A-dependent endonuclease of mouse, rabbit, or human cells 100-300 times less effectively than parent p5'A2'p5'A2'p5'A. In extracts of human Daudi cells where the monophosphate p5'A2'p5'A2'p5'A was able to effect ribosomal RNA cleavage at 2 X 10(-7) M, 14 required a concentration of 2 X 10(-5) M to bring about discernible rRNA cleavage. PMID- 2993625 TI - Immunoregulation by opioid peptides: I. Absence of classical opioid receptor on human mononuclear cells. AB - Potent effects of opiates and opioid peptides on the immune responsiveness of mononuclear cells have recently been described. The mechanism by which the effects are mediated was explored by examining the opioid receptor properties of these cells. Human mononuclear cells (B and T lymphocytes and monocytes) were examined for their ability to bind 3H-naloxone, a mu opioid ligand, and 3H-D-Ala2 D-Leu5-enkephalin, a delta opioid ligand. Both ligands bound to nonadherent cells as well as monocytes in a non-specific fashion and could not be competitively displaced by the addition of unlabeled compound. Cells which were stimulated in culture with mitogens did not develop receptors which bound either ligand in a specific fashion. Homogenization of human mononuclear cells prior to measuring binding significantly reduced non-specific binding of radioligands. Although under these conditions a limited amount of net specific binding was observed, this binding was not concentration dependent or saturable and varied considerably between individuals. These observations suggest that opioid effects on immune responsiveness are not mediated through a mononuclear opioid receptor. PMID- 2993626 TI - Modulation of lymphocyte adrenergic receptors by hormones and anti-asthmatic drugs. AB - Viable lymphocytes were used to investigate the binding of physiological concentrations of hormones and therapeutic concentrations of anti-asthmatic drugs to [3H]dihydroalprenolol (DHA, beta antagonist) and [3H]prazosin (alpha antagonist) receptor sites. All studied drugs and hormones inhibited the specific binding of prazosin to receptors identified with 10(-4)M phentolamine. Similar concentrations of drugs and hormones, with the exception of prostaglandin F2 alpha and cimetidine, inhibited the specific binding of DHA to lymphocyte receptors identified with 10(-7)M propranolol. These results confirm current mechanisms of hormone and drug action in asthma. PMID- 2993627 TI - Delta endotoxin inhibits Rb+ uptake, lowers cytoplasmic pH and inhibits a K+ ATPase in Manduca sexta CHE cells. AB - Delta endotoxin, a 68 kilodalton protein isolated from Bacillus thuringiensis spp. Kurstaki, is a potent entomocidal agent that alters a K+ current across midgut tissue of many phytophagous insects. This toxin completely inhibited the vanadate-sensitive 86Rb+ uptake and mimicked the vanadate-induced decrease in cytosolic pH in a cell line (CHE) originating from Manduca sexta embryonic tissue. The toxin also inhibited a K+-sensitive-ATPase in the plasma membranes isolated from these cells. Using the K+-sensitive-ATPase substrate p-nitrophenyl phosphate, delta endotoxin was found to have a Ki of 0.4 microM. These data suggest that the toxin inhibits a K+-ATPase responsible for 86Rb+ uptake in the CHE cells. The relationship between the toxin inhibition of K+-ATPase and toxin altered K+ current is discussed. PMID- 2993628 TI - Effect of metabolic depletion on the furosemide-sensitive Na and K fluxes in human red cells. AB - We report in this paper the effect of metabolic depletion on several modes of furosemide-sensitive (FS) Na and K transport in human red blood cells. The reduction of ATP content below 100 mumol/liter cells produced a marked decrease in the maximal activation (Vmax) of the outward, FS transport of Na and K into choline medium in the presence of ouabain (0.1 mM) and 1 mM MgCl2. The K0.5 for internal Na to activate the FS Na efflux was not altered by metabolic depletion. However, metabolic depletion markedly decreased the Ki for external K (Ko) to inhibit the FS Na efflux into choline medium (from 25 to 11 mM). Repletion of ATP content by incubation of cells in a substrate-rich medium recovered control levels of Vmax of the FS Na and K fluxes and of Ki for external K to inhibit FS Na efflux. The Vmax of FS Na and K influxes was also markedly decreased when the ATP content dropped below 100 mumol/liter cells. This was mainly due to a decrease in the inward-coupled transport of K and Na (NaO-stimulated K influx and the Ko-stimulated Na influx). The FS Ki/Ko exchange pathway of the Na-K cotransport, estimated from the FS K influx from choline-20 mM Ko medium into cells containing 22 mmol Na/liter cells, was also reduced by starvation. Starvation did not inhibit the FS Nai/Nao exchange pathway, estimated as FS Na influx from a medium containing 130 mM NaCl into cells containing 22 mmol Na/liter cells. The unidirectional FS 22Na efflux and influx were also measured in control and starved cells containing 22 mmol Na/liter cells, incubated in a Na medium (130 mM) at varying external K (0 to 20 mM). In substrate-fed cells, incubated in the absence of external K, FS Na efflux was larger than Na influx. This FS net Na extrusion (400 to 500 mumol/liter cells X hr) decreased when external K was increased, approaching zero around 15 mM Ko. In starved cells the net Na extrusion was markedly decreased and it approached zero at lower Ko than in substrate-fed cells. Our results indicate that the FS Na and K fluxes, and their major component, the gradient driven Na-K-Cl cotransport system, are dependent on the metabolic integrity of the cells. PMID- 2993629 TI - Proton exchange and base-pair kinetics of poly(rA).poly(rU) and poly(rI).poly(rC). AB - Proton exchange of poly(rA).poly(rU) and poly(rI).poly(rC) has been studied by nuclear magnetic resonance line broadening and saturation transfer from H2O. Five exchangeable peaks are observed. They are assigned to the imino, amino and 2'-OH ribose protons. The aromatic spectrum is also assigned. Contrary to previous observations, we find that the exchange of the imino proton is strongly buffer sensitive. This property is used to derive the base-pair lifetime, which is in the range of milliseconds at 27 degrees C, 100 times smaller than published values. The enthalpy for the base-opening reaction (-86 kJ/mol) and the insensitivity of the reaction to magnesium suggest that the open state involves a small number of base-pairs. The similarities in the exchange from the two duplexes indicate that the same open state is responsible for exchange of purine and pyrimidine imino protons. For the lifetime of the open state and for the base pair dissociation constant, we obtain only lower limits. At 27 degrees C they are three microseconds and 10(-3), respectively. The analysis that yields the much larger values published previously is based on the assumption that amino protons exchange only from open base-pairs. But theory and preliminary experiments indicate that it may occur from the closed duplex. The exchange of amino protons is slower than that of the imino protons. Exchange of the 2'-OH protons from the duplexes is much slower than from single-stranded poly(rU), and it is accelerated by magnesium. This could indicate hydrogen-bonding to backbone phosphate. Discrepancies between our results and those of previous studies are discussed. PMID- 2993630 TI - DNase I and nuclease S1 sensitivity of the rabbit beta 1 globin gene in nuclei and in supercoiled plasmids. AB - We have examined the nuclease sensitivity of the 5' flanking region of the rabbit beta 1 globin gene in bone marrow nuclei and in supercoiled plasmids. A DNase I hypersensitive site was found about 100 base-pairs 5' to the cap site in bone marrow nuclei. S1 nuclease can introduce a specific double-strand cut in the DNA in the same region. The presence of the nuclease-hypersensitive region correlates with the active transcription of gene beta 1 in bone marrow. Treatment with nuclease S1 of a supercoiled plasmid containing 1400 base-pairs of 5' flanking sequences as well as part of the beta 1 gene reveals a major double-strand cut 400 base-pairs 5' to the cap site. This cut maps within a stretch of repeating dinucleotides (C-T)12 and does not correspond to the in vivo site. Introduction of an RsaI fragment containing the nuclease S1-hypersensitive site into plasmid pBR322 shows that this fragment alone is sufficient to generate the hypersensitive site. Deletion of that RsaI fragment from the beta 1 plasmid reveals another site 1300 base-pairs upstream. Further deletion of this secondary site uncovers numerous other sites, none of which corresponds to the site in nuclei. Chromatin reconstitution with plasmids carrying the 5' flanking region of beta 1 and histones is capable of suppressing the in vitro nuclease-S1 hypersensitive site at --400 but is incapable of generating the in vivo site at - 100. Fine analysis at the nucleotide level of the early events in the digestion with nuclease S1 shows that the enzyme attacks preferentially the sequence (G A)12 on the message complementary strand. The region of DNA containing the supercoil-dependent S1 site adopts at least three different conformations that can be resolved electrophoretically. These different conformations are detected in linear restriction fragments and may represent non-B DNA or unusual B-form DNA. PMID- 2993631 TI - Nucleotide sequence of the genes involved in phosphate transport and regulation of the phosphate regulon in Escherichia coli. AB - The pstA(=phoT), pstB and phoU genes are situated at 84 minutes on the Escherichia coli genetic map. All of them are involved in the negative regulation of the phosphate regulon, and all of them except for phoU are required for the binding-protein-mediated, highly specific phosphate transport. We have determined the DNA sequence of about 4 X 10(3) bases of chromosomal segment containing these genes. Four translational reading frames (TRFs) were detected in the region. We attempted to assign the TRFs to the mutant alleles. Plasmids were constructed so that each contained only one of the TRFs, downstream from the lac promoter, to be used for the complementation tests. By this test, TRF-2, TRF-3 and TRF-4 were identified with the pstA(=phoT), pstB and phoU genes, respectively. Alkaline phosphatase-constitutive mutations of the two strains in our collection were complemented by the plasmid with the TRF-1 region. Therefore, we propose to designate the allele phoW. The order of the genes in this region has been established to be phoS-phoW-pstA(=phoT)-pstB-phoU counterclockwise on the E. coli genetic map. PMID- 2993632 TI - Crystal structure of Azotobacter cytochrome c5 at 2.5 A resolution. AB - The crystal structure of cytochrome c5 from Azotobacter vinelandii has been solved and refined to an R value of 0.29 at 2.5 A resolution. The structure of the oxidized protein was solved using a monoclinic crystal form. The structure was solved by multiple isomorphous replacements, re-fit to a solvent-leveled multiple isomorphous replacement map, and refined by restrained least squares. The structure reveals monomers associated about the crystallographic 2-fold axis by hydrophobic contacts at the "exposed heme edge". The overall conformation for the monomer is similar to that of Pseudomonas aeruginosa cytochrome c551. However, relative to a common heme conformation, c5 and c551 differ by an average of 6.8 A over 82 alpha-carbon positions and the propionates of c5 are much more exposed to solvent. The shortest heme--heme contact at the "dimer" interface is 6.3 A (Fe to Fe 16.4 A). Alignment of c5 and c551 shows that the two cytochromes, in spite of sequence differences, have remarkably similar charge distributions. A disulfide stacks on a tyrosine between the N- and C-terminal helices. PMID- 2993633 TI - Quantifying cell death in the myocardium: myosin specific antibody in the evaluation of membrane defects. AB - Methods for evaluating cardiac myocyte necrosis utilizing antibodies specific for the heavy or light chains of cardiac myosin are reviewed. Cell death, associated with sarcolemmal disruption, results in the leakage of myosin light chains from the cytoplasm as well as the accessibility of myosin heavy chains to exogenous specific antibodies. Measurement of plasma light chain concentration has been useful in the diagnosis of myocardial infarction, though more recently, patients with congestive cardiomyopathy associated with an inflammatory infiltrate have been identified by an elevated plasma light chain concentration. The binding of myosin heavy chains to necrotic myocytes has been useful in the study of mechanisms of ischemic cell death in cell culture, in the diagnosis and quantification of myocardial infarction, both experimentally and clinically, and more recently in the study of experimental myocarditis and cardiac transplantation. It is hoped that these methods may evolve as useful clinical tools in the identification of those cardiomyopathy patients whose course is characterized by rapid myocyte loss. PMID- 2993634 TI - Paraquat toxicity in vitro. I. Pulmonary alveolar macrophages. AB - When the herbicide paraquat (1,1'-dimethyl-4,4'-bipyridylium) was administered to adult rat pulmonary alveolar macrophages (PAM) in primary culture, both a time dependent and a dose-dependent cytotoxic response (cell death) was observed. An LD50 value of 1 mM was calculated when these cells were exposed to paraquat in vitro for 12 h in Ham's F12 culture medium at 30 degrees C. Cell death was accompanied by the formation of TBA-reactive substances (lipid peroxidation) and was potentiated by hyperoxia (95% O2). In a 95% O2-5% CO2 atmosphere, an LD50 value of 0.1 mM was calculated. In addition, the presence of superoxide dismutase in the culture medium (1700 units/ml) inhibited the cytotoxic response. Since [14C]paraquat was not absorbed into these cells, extracellular superoxide anion radical formation was investigated as the cause of the observed cell death. Paraquat (0.5 mM) was found to stimulate extracellular O-2 generation, from PAM, but only in nonactivated cells. A sevenfold enhancement over the resting rate of radical generation was observed in the presence of paraquat. No increase in the O 2 generation rate of activated macrophages was observed upon the addition of paraquat to the culture medium. These data indicate that paraquat is cytotoxic to the pulmonary alveolar macrophage and further suggest that this cytotoxicity is mediated, at least in part, by an excess, extracellular production of active oxygen species. Implications of these findings with respect to the currently accepted hypothesis of paraquat poisoning in vivo are discussed. PMID- 2993635 TI - Mutagenicity testing of ST-film with the Ames test, chromosome test in vitro and micronucleus test in female mice. AB - The mutagenicity of ST-film and its components polyoxyethylenenonylether (NP-10) and polyvinylalcohol (PVAL) were studied by a reverse mutation test in S. typhimurium (Ames test), a chromosome test in cultured Chinese hamster V 79 cells and a micronucleus test in female mice. The results obtained from these test systems were all negative. Thus, it may be concluded that ST-film is not mutagenic either in vitro or in vivo mutagenicity testing systems. PMID- 2993636 TI - Studies on the delayed neurotoxicity of organophosphorus compounds--(III). AB - TOCP (Tri-orthocresyl phosphate), an organophosphorus compound, has been implicated in producing neuropathy in the male S. D. rats. Repeated subcutaneous doses of TOCP (600 mg/kg) for up to 6 weeks produced ataxia, most striking at 50 days after final injection, followed by gradual recovery. Ultrastructurally, the internal structure of affected nerve fibers was primarily composed of altered smooth endoplasmic reticulum, tubular membrane system, and mitochondria, although myelin sheath was found to be essentially normal. In the histopathological examination, axonal and myelin degeneration was disclosed in the gracile nucleus and in the gracile fasciculus of the cords as well as in the sciatic nerves. The localization and degree of these changes were considered to be "dying back", showing systemic neuropathy. In addition, muscular lesion showed small group atrophy, corresponding to Type I fiber atrophy. PMID- 2993637 TI - Twenty-eight week toxicity study of perlite powder in mice. AB - Groups of 21 male and 21 female mice were given diets containing 20%, 10%, 1% or 0% (control) perlite for 28 weeks. Appearance, behavior, mortality, and food consumption of mice of treated groups were not affected during the experimental period. Males of the 20% and 10% groups did experience slightly reduced growth rate. No significant drug-related changes were found in urinalysis, hematology, serum chemistry, and organ weight. No differences were found between control and treated groups in autopsy and histopathological findings. Therefore, the maximum no-effect level is considered to be 1% in the diet which developed no toxic changes in any items examined in mice treated orally with perlite for 28 weeks. PMID- 2993638 TI - Effect of oxygen and superoxide anion on mutation rate in Ames test. AB - Effects of oxygen and superoxide anion to mutation rate were examined using the method of Ames. As test samples, 3-methylcholanthrene or 3,4-benzpyrene dissolved in squalene was used. In order to increase the concentration of dissolved oxygen in the medium during preincubation, oxygen was bubbled into the medium. Various combinations of xanthine oxidase, superoxide dismutase and hypoxanthine were used with an intent of changing the concentration of superoxide anion in the medium during the preincubation. An increase in the concentration of dissolved oxygen and superoxide anion resulted in a lower mutation rate, clearly showing the relationships of mutation rate with oxygen and with superoxide anion. PMID- 2993640 TI - Serial changes in sonographic appearance after transcatheter hepatic arterial embolization. AB - Serial changes in sonographic appearance after transcatheter hepatic arterial embolization (TAE) were studied in 22 patients who had hepatoma and one patient who had a hepatoblastoma. These changes were classified into three types. In cases of type 1, the internal echo of the entire tumor became remarkably echogenic with or without an acoustic shadow. In cases of type 2, echogenic areas or scattered echogenic spots with or without acoustic shadows were observed. In Type 3 cases, no changes in sonographic appearance were observed. Tumor diameters in all of the type 1 cases were less than 4.9 cm. Various size tumors were observed in the cases of type 2 and type 3. Transcatheter arterial embolization was completely effective or effective in 89 per cent of the type 1 and type 2 cases, but was not effective in 75 per cent of the type 3 cases. The sonographic changes correlated well with the effectiveness of TAE. PMID- 2993639 TI - Preliminary clinical experience with the use of absorbable mesh splenorrhaphy. AB - Splenorrhaphy was performed in six patients using absorbable polyglycolic acid mesh. The sustained injuries included blunt and penetrating trauma and varied from through-and-through puncture with hematoma and deep lacerations to amputation of a portion of the spleen. A number of different splenorrhaphy techniques were required. In all six cases, the procedure was accomplished without abscess formation, postoperative bleeding, or complications related to the use of the mesh. PMID- 2993641 TI - Regulation of herpes simplex virus-specific lymphoproliferation by suppressor cells. AB - We investigated the regulation of the herpes simplex virus (HSV)-specific lymphoproliferative response (LPR) by suppressor cells. The chief cell types in HSV-immune splenocytes proliferating in response to the antigen were Lyt 1+ and Lyt 2+ T cells, which accounted for approximately 60 and 40% of the response, respectively. Because the total responsiveness of splenocytes was enhanced after depletion of Lyt 2+ cells, the LPR was assumed to be subject to regulation by an Lyt 2+ suppressor cell. This was shown to be the case with an experimental design in which suppressor cell activity was induced in one culture, the cells were irradiated, and the effects on LPR were measured in a test antigen-stimulated culture. The cell responsible for suppression was shown to be Lyt 2+ IJ+, and the actual suppressor effect was not antigen specific. Cellular requirements for the generation of suppression were also investigated. The three distinct cell types that appeared to be required were Lyt 2+ and Lyt 1+ T cells and an IJ+ antigen presenting cell. Of the three cell types, only the Lyt 2+ cell needed to be from HSV-immune animals. The implications of our model system for the better understanding of the role of immunity in herpesvirus pathogenesis are discussed. PMID- 2993642 TI - Origin binding by a 100,000-dalton super-T antigen from SVT2 cells. AB - The SVT2 line of simian virus 40-transformed mouse cells expresses little or no wild-type-size A protein (T antigen). Instead, a variant form is produced in these cells that is larger than normal-size A protein. This variant form has an Mr of 100,000 (100K super-T antigen) and is found primarily in complexes with the host-cell-coded p53 protein. Binding of the 100K super-T antigen to simian virus 40 origin region DNA was assayed by immunoprecipitation of super-T antigen-DNA complexes and then digestion with DNase I. DNA sequences associated with super-T antigen were protected from digestion and retained in the immune complex, while unprotected sequences were digested and released. The 100K super-T antigen efficiently protects DNA sequences in the previously defined regions I and II (P. Tegtmeyer, B. A. Lewton, A. L. DeLucia, V. G. Wilson, and K. Ryder, J. Virol. 46:151-161, 1983). Within region II (the origin of replication), the pattern and size of protected fragments are identical for super-T antigen and purified wild type A protein. Thus, even though super-T antigen is larger than wild-type A protein, both must bind with the same alignment on origin DNA. Furthermore, complexes between the host-cell-coded p53 protein and the 100K super-T antigen also retain the ability to bind in regions I and II. PMID- 2993643 TI - Sialylated oligosaccharides O-glycosidically linked to glycoprotein C from herpes simplex virus type 1. AB - Glycoprotein C (gC) was purified by immunoabsorbent from herpes simplex virus type-1-infected BHK cells labeled with [14C]glucosamine for 11 h and chased for 3 h. Glycopeptides obtained by pronase digestion of gC were fractionated by Bio-Gel filtration and concanavalin A-Sepharose chromatography. Each glycopeptide fraction was analyzed for amino sugar composition by thin-layer chromatography. The majority of radioactivity was recovered as N-acetylglucosamine, but a significant amount of labeled N-acetylgalactosamine was detected and recovered preferentially in some glycopeptide species. Mild alkaline borohydride treatment of the glycopeptides resulted in the release of small degradation products which contained N-acetylgalactosaminitol as the major labeled component and a drastic reduction of N-acetylgalactosamine in the residual glycopeptides. These results demonstrated that gC carries O-glycosidically linked oligosaccharides in addition to the N-linked di- and triantennary glycans previously described (F. Serafini Cessi, F. Dall'Olio, L. Pereira, and G. Campadelli-Fiume, J. Virol. 51:838-844, 1984). Chromatographic behavior on DEAE-Sephacel chromatography and neuraminidase digestion of O-linked oligosaccharides indicated the presence of two major sialylated species carrying one and two sialic acid residues, respectively. The characterization of a peculiar glycopeptide species supported the notion that some of the O-linked oligosaccharides are bound to a cluster of hydroxyamino acids located near an N-glycosylation site which carries one N-linked diantennary oligosaccharide. PMID- 2993644 TI - Regulation of cytomegalovirus gene expression: alpha and beta promoters are trans activated by viral functions in permissive human fibroblasts. AB - We have fused immediate (alpha) and delayed (beta) early promoter-regulatory sequences taken from the cytomegalovirus (CMV) genome to Escherichia coli lacZ (beta-galactosidase) as an indicator gene to study regulated expression of these promoters. After transfection of human fibroblast cells with plasmid constructs carrying beta-galactosidase fusions, and subsequent infection with CMV, we have demonstrated that viral trans-acting functions up-regulate the expression of these genes in a temporally authentic manner. The alpha promoter is activated even when de novo protein synthesis is blocked and when UV-inactivated virus is used, suggesting that, as for herpes simplex virus type 1 (HSV-1), a virion structural protein is responsible for its up-regulation. We have found that HSV 1, as well as CMV, is capable of trans activating the CMV alpha promoter. The beta promoter is activated by CMV but is completely unresponsive to HSV-1 infection. The temporal synthesis of the alpha and beta promoters in the transient expression system conforms with their natural regulation during viral replication. The beta-galactosidase fusions we describe provide a most exquisitely sensitive indicator system for the study of cis- and trans-acting viral regulatory functions. PMID- 2993645 TI - Nucleotide sequence and biochemical activities of the Moloney murine sarcoma virus strain HT-1 mos gene. AB - The nucleotide sequence of the Moloney murine sarcoma virus strain HT-1 (HT1MSV) mos gene differs from that of the cellular mos gene in three positions, but these are silent changes, and the amino acid sequence of the v-mos and c-mos open reading frames are identical. We have overproduced the mos HT1MSV (equivalent to c-mos) in Escherichia coli under the control of phage lambda promoter (pL). The E. coli p40mos protein thus obtained was partially purified and examined for several biochemical activities. We show that the p40mos binds ATP analog p fluorosulfonylbenzoyladenosine and exhibits ATPase activity. PMID- 2993646 TI - Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells. AB - A recombinant baculovirus containing the Escherichia coli chloramphenicol acetyltransferase (CAT) gene under the control of the Rous sarcoma virus long terminal repeat promoter and the E. coli beta-galactosidase gene under the control of the very late baculoviral polyhedrin promoter was used to determine if Autographa californica nuclear polyhedrosis virus, a baculovirus of Lepidoptera, can enter and express viral DNA in dipteran (Drosophila sp.) and mammalian (Mus sp.) cells that are considered refractory to baculovirus replication. Following infection, CAT gene expression was observed in both dipteran and mammalian cells, but expression in the mammalian cell line was less than 0.05% of that observed in either dipteran or lepidopteran cells. Although the level of CAT gene expression was similar in permissive lepidopteran and nonpermissive dipteran cells, expression of beta-galactosidase activity from the late polyhedrin promoter in dipteran or mammalian cells was less than 0.3% of the levels observed in lepidopteran cells. These results indicate that foreign gene expression in nonpermissive cells is promoter dependent and that late viral gene expression is restricted in these cells. The Rous sarcoma virus long terminal repeat allows substantial CAT gene expression in both a D. melanogaster cell line and Aedes aegypti midgut cells. Baculovirus DNA undergoes a limited number of replications in Drosophila cells. The results are relevant to baculovirus host range, the safety of baculoviruses as pesticides, and the development of baculovirus pesticides with expanded host ranges. PMID- 2993648 TI - Chromosomal assignment of two endogenous ecotropic murine leukemia virus proviruses of the AKR/J mouse strain. AB - The AKR/J mouse strain is genetically fixed for three different ecotropic murine leukemia virus genomes, designated Akv-1, Akv-3, and Akv-4 (Emv-11, Emv-13, and Emv-14). With recombinant inbred strains and crosses with linkage-testing stocks, Akv-3 and Akv-4 were placed on the mouse chromosome map. Akv-3, which encodes a replication-defective provirus, maps near the agouti coat color locus, a, on chromosome 2. Akv-4, which is replication competent, maps near the neurological mutant gene locus trembler, Tr, on chromosome 11. Akv-1 and Akv-2 (Emv-12), an ecotropic provirus carried by AKR/N but not AKR/J, have previously been mapped to chromosome 7 and 16, respectively. Thus, the four Akv proviruses mapped to date are on four different chromosomes. Akv-3 is the second ecotropic murine leukemia virus provirus to be mapped near the agouti locus. The results are discussed in relation to possible nonrandomness of viral integration. PMID- 2993647 TI - Evidence for the presence of an inhibitor on ribosomes in mouse L cells infected with mengovirus. AB - After infection of mouse L cells with mengovirus, there is a rapid inhibition of protein synthesis, a concurrent disaggregation of polysomes, and an accumulation of 80S ribosomes. These 80S ribosomes could not be chased back into polysomes under an elongation block. The infected-cell 80S-ribosome fraction contained twice as much initiator methionyl-tRNA and mRNA as the analogous fraction from uninfected cells. Since the proportion of 80S ribosomes that were resistant to pronase digestion also increased after infection, these data suggest that the accumulated 80S ribosomes may be in the form of initiation complexes. The specific protein synthetic activity of polysomal ribosomes also decreased with time of infection. However, the transit times in mock-infected and infected cells remained the same. Cell-free translation systems from infected cells reflected the decreased protein synthetic activity of intact cells. The addition of reticulocyte initiation factors to such systems failed to relieve the inhibition. Fractionation of the infected-cell lysate revealed that the ribosomes were the predominant target affected. Washing the infected-cell ribosomes with 0.5 M KCI restored their translational activity. In turn, the salt wash from infected-cell ribosomes inhibited translation in lysates from mock-infected cells. The inhibitor in the ribosomal salt wash was temperature sensitive and micrococcal nuclease resistant. A model is proposed wherein virus infection activates (or induces the synthesis of) an inhibitor that binds to ribosomes and stops translation after the formation of the 80S-ribosome initiation complex but before elongation. The presence of such an inhibitor on ribosomes could prevent them from being remobilized into polysomes in the presence of an inhibitor of polypeptide elongation. PMID- 2993650 TI - Inversion and circularization of the varicella-zoster virus genome. AB - The genome of varicella-zoster virus (VZV) is a linear, double-stranded molecule of DNA composed of a long (L) region covalently linked to a short (S) region. The S region is capable of inverting relative to a fixed orientation of the L region, giving rise to two equimolar populations. We have investigated other forms of the VZV genome which are present in infected cells and packaged into nucleocapsids. That a small proportion of nucleocapsid DNA molecules also possess inverted L regions has been verified by the identification of submolar restriction fragments corresponding to novel joints and novel ends generated by such an inversion. The presence of circular molecules has been investigated by agarose gel electrophoresis. Bands corresponding to circular forms were present in small amounts in both VZV-infected cell DNA and nucleocapsid DNA. Southern blot analysis verified that these bands contained VZV sequences. We therefore conclude that the VZV genome may occasionally contain an inverted L region or exist in a circular configuration. PMID- 2993649 TI - Virus-induced modification of the host cell is required for expression of the bacterial chloramphenicol acetyltransferase gene controlled by a late herpes simplex virus promoter (VP5). AB - The requirements for expression of genes under the control of early (alkaline exonuclease) and late (VP5) herpes simplex virus type 1 (HSV-1) gene promoters were examined in a transient expression assay, using the bacterial chloramphenicol acetyltransferase gene as an expression marker. Both promoters were induced, resulting in the production of high levels of the enzyme upon low multiplicity infection by HSV-1. S1 nuclease analysis of hybrids between RNA isolated from infected cells containing HSV-1 promoter constructs and marker gene DNA demonstrated normal transcriptional initiation of the marker gene directed by the viral promoters. Viral DNA sequences no more than 125 bases 5' of the putative transcriptional cap site were sufficient for maximum activity of the late promoter. In contrast to expression controlled by the early gene, the late promoter was not active at a measurable level in uninfected cells until DNA sequences between 75 and 125 bases 5' of the transcriptional cap site were deleted. Cotransfection of cells with the expression marker controlled by HSV promoters and a cosmid containing HSV alpha (immediate-early) genes indicated that full expression of both early and late promoters requires the same virus induced host cell modifications. Inhibition of viral DNA synthesis results in an increased rate of transient expression of marker genes under control of either early or late promoters in contrast to the situation in normal virus infection. These data provide evidence that the normal course of expression of late HSV genes involves negative modulation of potentially active promoters in the infected cell. PMID- 2993651 TI - Application of antibody to synthetic peptides for characterization of the intact and truncated alpha 22 protein specified by herpes simplex virus 1 and the R325 alpha 22- deletion mutant. AB - The alpha 22 protein is one of five proteins synthesized immediately after infection of permissive cells with herpes simplex virus 1 and 2 (HSV-1 and HSV 2). On the basis of the reported nucleotide sequence of the HSV-1 gene, we synthesized two peptides containing the predicted amino acids 12 through 23 (12 residues) and 21 through 36 (16 residues) in two hydrophilic domains near the N terminus of the protein. Rabbit antisera made against these peptides were then used to characterize the alpha 22 protein made by wild-type HSV-1(F) strain and by an HSV-1 mutant, R325, carrying a 500-base-pair deletion within the coding domain of the gene. The results were as follows. (i) Both antisera reacted with HSV-1(F) alpha 22 protein in lysates electrophoretically separated in denaturing polyacrylamide gels and electrically transferred to a nitrocellulose sheet; neither antiserum reacted with the corresponding HSV-2 protein. The protein accumulated at 34 and 39 degrees C in the nucleus of infected permissive HEp-2 and baby hamster kidney (BHK) cells. The protein formed at least five spots differing in charge, mobility, and extent of phosphorylation on two-dimensional electrophoretic separation. (ii) The antisera reacted with a truncated nuclear protein (33,700 apparent molecular weight) in permissive HEp-2 and restrictive BHK cells infected with R325 and incubated at 39 degrees C but not at 34 degrees C. The truncated protein represents, therefore, the product of the undeleted 5' domain of the alpha 22 gene in R325. (iii) The presence of identical as well as slower migrating, reactive proteins in infected BHK cell lysates indicated that wild-type and truncated alpha 22 proteins are processed differently in BHK and HEp-2 cells. PMID- 2993654 TI - Concerted DNA rearrangements in Moloney murine leukemia virus-induced thymomas: a potential synergistic relationship in oncogenesis. AB - Rat thymic lymphomas induced by Moloney murine leukemia virus carry DNA rearrangements due to provirus integration in at least five independent cellular DNA domains (Mlvi-1, Mlvi-2, Mlvi-3, RMoInt-1, and c-myc). We had previously shown that rearrangements in more than one of these domains could occur in the same tumor. In this report we extend these findings by showing that, with one exception, tumors containing provirus insertions in Mlvi-1 always contained provirus insertions in a second locus, Mlvi-2. To determine whether both events occurred in the same population of tumor cells, we examined the clonal nature of these tumors by taking advantage of allelic polymorphisms that occur naturally in both Mlvi-1 and Mlvi-2. Tumors with provirus insertions in both Mlvi-1 and Mlvi-2 arising in rats heterozygous at one of these loci were identified. DNA from these tumors was analyzed by restriction endonuclease digestion and hybridization to DNA probes derived from both Mlvi-1 and Mlvi-2. Thus, we determined the clonal nature of three thymomas and showed that in these tumors both insertion events occurred in the same population of tumor cells. The concomitant appearance of provirus insertions in Mlvi-1 and Mlvi-2 suggests a synergism of these two events that may be important in tumor induction and progression. PMID- 2993653 TI - Biologically active proviral clone of myeloblastosis-associated virus type 1: implications for the genesis of avian myeloblastosis virus. AB - A biologically active myeloblastosis-associated virus (MAV) provirus was cloned from a bacteriophage recombinant library constructed from leukemic chicken myeloblast DNA. The restriction endonuclease map of this clone was consistent with that of a type 1 MAV (MAV-1). Interference assays of virus recovered from cultured chicken embryo fibroblasts after DNA transfection established that the provirus was infectious and confirmed that it belonged to avian retrovirus subgroup A (type 1). Antipeptide antibodies raised against the env-encoded carboxyl terminus of p48myb, the transforming protein of avian myeloblastosis virus, specifically immunoprecipitated the gp37env from quail cells transfected with MAV-1 proviral DNA but not from cells infected with MAV-2. This suggests that MAV-1 rather than MAV-2 is the progenitor helper virus from which avian myeloblastosis virus arose by the transduction of cellular proto-oncogene sequences. PMID- 2993652 TI - Segregation of restriction fragment length polymorphism in an interspecies cross of laboratory and wild mice indicates tight linkage of the murine IFN-beta gene to the murine IFN-alpha genes. AB - Southern blot analysis with murine (Mu) interferon (IFN)-alpha cDNA of restricted genomic DNA of three inbred strains of mice belonging to the species Mus musculus domesticus (BALB/c, C57BL/6, and DBA/2) revealed only a limited degree of polymorphism. For example, with HindIII there were only two polymorphic bands out of 14 hybridizing fragments. With Mu IFN-beta cDNA there was no polymorphism at all between BALB/c and C57BL/6 in DNA restricted with seven different enzymes. In contrast, HindIII-restricted DNA of an inbred strain of wild mice (M. spretus Lataste) hybridized with the IFN-alpha probe displayed a high degree of polymorphism compared with the three strains of laboratory mice and was also polymorphic when probed with IFN-beta cDNA. Although M. musculus domesticus and M. spretus Lataste represent different species, certain interspecies crosses are possible in the laboratory. This enabled us to follow segregation of restriction fragment length polymorphism in HindIII-restricted DNA obtained from 18 backcross progeny of a (DBA/2 X M. spretus)F1 X DBA/2 interspecies cross. There was complete coincidence between the segregation of parental (DBA/2) and (DBA/2 X M. spretus)F1-type IFN-beta and IFN-alpha restriction fragment length polymorphism, indicating tight linkage of the IFN-beta and IFN-alpha genes. In addition, in 15 of 18 progeny the segregation coincided with that of the brown locus on chromosome 4, in accord with previous results obtained with the IFN-alpha probe in strains derived from crosses between BALB/c and C57BL/6 mice. Thus, the Mu IFN beta gene is tightly linked to the Mu IFN-alpha gene cluster on chromosome 4 near the brown locus. PMID- 2993655 TI - Isolation of human cytomegalovirus intranuclear capsids, characterization of their protein constituents, and demonstration that the B-capsid assembly protein is also abundant in noninfectious enveloped particles. AB - Two types of intranuclear capsids have been recovered from human cytomegalovirus (HCMV, strain AD169)-infected cells. By analogy with strain Colburn (simian CMV) particles, these have been designated as A- and B-capsids. Both types of capsids are composed of proteins with molecular weights of 153,000 (major capsid protein), 34,000 (minor capsid protein), 28,000, and 11,000 (smallest capsid protein). In addition to these species, B-capsids contain a 36,000-molecular weight (36K) protein which has been designated as the HCMV "assembly protein," based on its similarities to counterparts in strain Colburn CMV (i.e., 37K protein) and herpes simplex virus (i.e., VP22a/p40/NC-3/ICP35e). Peptide comparisons established that the assembly protein of HCMV B-capsids and the 36K protein that distinguishes HCMV noninfectious enveloped particles from virions are the same, providing direct evidence that noninfectious enveloped particles are enveloped B-capsids. PMID- 2993656 TI - A mos oncogene-containing retrovirus, myeloproliferative sarcoma virus, transforms rat thyroid epithelial cells and irreversibly blocks their differentiation pattern. AB - Differentiated, cloned rat thyroid epithelial cells (424 cells) were infected with a wild-type and a temperature-sensitive strain of the myeloproliferative variant of the Moloney murine sarcoma virus. The thyroid cells were productively infected and transformed by both virus strains and displayed some of the typical properties of malignant cells, such as morphological changes, growth in soft agar, and in vivo tumorigenicity. The acquisition of the transformed phenotype by the virus-infected cells was accompanied by a loss of the typical differentiated features of the thyroid epithelium, such as thyroglobulin (TG) secretion, iodide uptake, and dependence for growth on six factors including thyrotropin, the physiological thyroid stimulator. TG mRNA could not be demonstrated in cells transformed by both viral strains, suggesting a block at the level of the TG gene transcription. While the transformed state of the cell clones infected with the temperature-sensitive strain could be reverted by shifting the cultures to the temperature nonpermissive for transformation (39 degrees C), no reversion of the differentiated functions took place after such a shift, showing that the v-mos oncogene irreversibly shuts off the differentiation of thyroid epithelial cells in vitro. These results demonstrate, for the first time, an oncogenic potential of the v-mos oncogene family towards differentiated epithelial cells in vitro. PMID- 2993657 TI - Formation of a cruciform structure at the simian virus 40 replication origin abolishes T-antigen binding to the origin in vitro. AB - Heteroduplex DNA molecules were formed by annealing an intact simian virus replication origin-containing fragment to a mutant derivative lacking the indigenous wild-type 27-base-pair (bp) inverted repeat within this structure and containing a nonhomologous 26-bp inverted repeat sequence in its place. Results of restriction enzyme and S1 endonuclease cleavage analyses strongly suggested that a 13-bp stem-loop structure formed at the site of nonhomology between these two DNAs. This structure lies within the boundary of simian virus 40 T-antigen binding site 2, and its presence inhibited T-antigen binding to that sequence but not to an adjacent higher-affinity binding site (site 1). Therefore, the conformation of sequences within an otherwise intact T-antigen-binding site can have major effects upon T-antigen binding there. PMID- 2993658 TI - Proteins associated with human parainfluenza virus type 3. AB - The polypeptides associated with human parainfluenza virus type 3 were identified. Five proteins were present in detergent- and salt-resistant viral cores. Of these, three proteins designated NP0, NP1, and NP2 of 68,000, 58,000, and 52,000 daltons, respectively, were stably associated with 50S RNA in CsCl gradient-purified nucleocapsids. The amounts of NP1 and NP2 were variable, and these proteins were shown to be structurally related to the major nucleocapsid protein (NP0) by partial Staphylococcus aureus V8 protease mapping. The other core proteins included a 240K protein designated L (candidate for the viral polymerase) and an 84K protein designated as the phosphoprotein (P) on the basis of a predominant incorporation of Pi. The viral envelope had four prominent proteins (72, 53, 40, and 12K) under reducing conditions of electrophoresis. The 72 and 53K proteins were specifically labeled with [3H]glucosamine and [3H]mannose. When sulfhydryl reagents were removed, a new 62K protein was visualized in place of the 72, 53, and 12K proteins. The 53 and 12K proteins were interpreted to be the two subunits (F1 and F2) of the fusion protein, and the 72K protein was designated as the HN (hemagglutinin-neuraminidase) glycoprotein. The unglycosylated 40K protein represented the viral matrix protein (M). Immunoprecipitation of infected cell lysates with rabbit hyperimmune antiserum against purified virus confirmed the viral origin of these polypeptides. PMID- 2993660 TI - Virion component of herpes simplex virus type 1 KOS interferes with early shutoff of host protein synthesis induced by herpes simplex virus type 2 186. AB - Herpes simplex virus (HSV) strains HSV type 1 (HSV-1) KOS and HSV-2 186 are representative of delayed and early shutoff strains, respectively, with regard to their ability to inhibit protein synthesis in Friend erythroleukemia cells. When these cells were simultaneously infected with HSV-1 KOS and HSV-2 186, HSV-1 KOS interfered with the rapid suppression of globin synthesis induced by HSV-2 186. The observed interference was competitive and not due to exclusion of HSV-2 by HSV-1 at the level of adsorption. Furthermore, UV-irradiated HSV-1 KOS was also effective at interfering with the early shutoff function of HSV-2 186, indicating that a virion component is responsible for the observed interference. PMID- 2993659 TI - Primary structure of p19 species of avian sarcoma and leukemia viruses. AB - The internal structural proteins of avian sarcoma and leukemia viruses are derived from a precursor polypeptide that is the product of the viral gag gene. The N-terminal domain of the precursor gives rise to p19, a protein that interacts with the lipid envelope of the virus and that may also interact with viral RNA. The C terminus of p19 from the Prague C strain of Rous sarcoma virus was previously assigned to a tyrosine residue 175 amino acids from the N terminus. We have used metabolic labeling and carboxypeptidase digestion to show that the C terminus of p19 is actually tyrosine 155. This implies the existence of a sixth gag protein 22 amino acids in length and located between p19 and p10 on the gag precursor. The p19 species of some recombinant avian sarcoma viruses and of the defective endogenous virus derived from the ev-1 locus migrate on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as if they were about 4,000 daltons smaller than p19. We have elucidated the structure of these forms, called p19 beta, by analysis of the proteins and determination of the DNA sequence of the p19 region of the gag gene from ev-1 and ev-2. Esterification of carboxyl groups completely suppressed the differences in migration of p19 and p19 beta. Peptide mapping showed the altered mobility to be determined by sequences in the C-terminal cyanogen bromide fragment of the proteins. We conclude from the DNA sequence that a single glutamate-lysine alteration is responsible for the altered electrophoretic mobility. PMID- 2993661 TI - Demonstration of an immunodominant neutralization site by analysis of antigenic variants of SA11 rotavirus. AB - Serotype-specific monoclonal antibodies were used to select mutants of SA11 rotavirus that were resistant to neutralization. The antigenic characteristics of these mutants were studied with with a panel of monoclonal antibodies. We isolated one type of mutant which showed a dramatic increase (greater than 10 fold) in resistance to neutralization by hyperimmune antiserum, and this together with other data indicates the presence on the rotavirus major outer shell glycoprotein of an immunodominant antigenic site involved in virus neutralization. The mutants were also useful in classifying neutralizing monoclonal antibodies. PMID- 2993662 TI - Lack of evidence for methylation of parental and newly synthesized adenovirus type 2 DNA in productive infections. AB - Methylations of highly specific sites in the promoter and 5' regions of eucaryotic genes have been shown to shut off gene activity and thus play a role in the long-term regulation of gene expression. It was therefore of interest to investigate whether site-specific DNA methylations could also play a role in adenovirus DNA in productive infections. It has been reported earlier that adenovirion DNA is not detectably methylated (U. Gunthert, M. Schweiger, M. Stupp, and W. Doerfler, Proc. Natl. Acad. Sci. USA 73:3923-3927, 1976). In the present study, evidence for the methylation of cytidine residues in 5'-CCGG-3' and 5'-GCGC-3' sequences or the methylation of adenine residues in 5'-GATC-3' and 5'-TCGA-3' sequences in intranuclear adenovirus type 2(Ad2) DNA isolated and analyzed early (5 h) or late (24 h) after infection could not be obtained. In Ad2 DNA, 22.5% of all 5'-CG-3' dinucleotides reside in 5'-CCGG-3' and 5'-GCGC-3' sequences. Intranuclear viral DNA was examined by restriction endonuclease cleavage by using HpaII, MspI, HhaI, DpnI, or TaqI and Southern blot hybridizations. The HindIII fragments of Ad2 DNA served as hybridization probes. The data rendered it very unlikely that free intracellular adenovirus DNA in productively infected cells was extensively methylated. Thus, DNA methylation was not a likely element in the regulation of free adenovirus DNA expression in productively infected cells. PMID- 2993663 TI - Interaction of simian virus 40 small-T antigen produced in bacteria with 56K and 32K proteins of animal cells. AB - Small-t antigen produced in bacteria interacted with two animal cell proteins with molecular weights of 56,000 and 32,000, as did the viral antigen from infected cells. Demonstration of this specific interaction required the enrichment of native, monomeric small-t antigen from extracts in which much of the small-t antigen was highly aggregated. PMID- 2993664 TI - Random association of Epstein-Barr virus genomes with host cell metaphase chromosomes in Burkitt's lymphoma-derived cell lines. AB - The random association of Epstein-Barr virus DNA with host cell metaphase chromosomes of all sizes in Burkitt's lymphoma-derived cell lines was demonstrated by two substantially different techniques, namely fluorescence activated chromosome sorting and in situ hybridization. The nature and potential importance of this association are discussed. PMID- 2993665 TI - Cross-reactivity between herpes simplex virus glycoprotein B and a 63,000-dalton varicella-zoster virus envelope glycoprotein. AB - Cross-reactive monoclonal antibodies recognizing both herpes simplex virus (HSV) glycoprotein B and a major 63,000-dalton varicella-zoster virus (VZV) envelope glycoprotein were isolated and found to neutralize VZV infection in vitro. None of the other VZV glycoproteins was recognized by any polyclonal anti-HSV serum tested. These results demonstrate that HSV glycoprotein B and the 63,000-dalton VZV glycoprotein share antigenic epitopes and raise the possibility that these two proteins have a similar function in infection. PMID- 2993666 TI - Different forms of membrane-associated herpes simplex virus glycoproteins induce functionally distinct subsets of herpes simplex virus-specific suppressor T cells. AB - Previous studies have shown that two types of virus-specific suppressor T cells (Ts) are induced in mice made tolerant with herpes simplex virus (HSV)-infected spleen cells (SC). One type of Ts blocks the afferent phase of the delayed hypersensitivity response to HSV (Ts-aff), and the other blocks the efferent or effector phase (Ts-eff). In this report we show that the induction requirements for these suppressor populations differ. Injection of SC infected for 6 h with HSV at a multiplicity of infection of 5 or less or treated with heat-inactivated virus induced only Ts-aff. Similar results were seen with SC incubated for 90 min in virus-free preparations containing only viral proteins. In contrast, the Ts eff population was induced only by SC treated for 6 h with infectious HSV at a multiplicity of infection of 10. Collectively, these data indicate that Ts-aff are induced by adsorbed HSV antigens on SC, whereas Ts-eff are induced by nascent HSV antigens expressed on infected SC. In addition to their induction requirements, the two types of regulatory cells differ in their expression of effector function. Ts-eff but not Ts-aff require a cyclophosphamide-sensitive target cell in the immune recipient for suppressor function. The possible identity of this target cell and the significance of the different induction requirements between the two types of Ts are discussed. PMID- 2993667 TI - Herpesvirus sylvilagus infects both B and T lymphocytes in vivo. AB - Herpesvirus sylvilagus infection of cottontail rabbits (Sylvilagus floridanus) was studied as a model of herpesvirus-induced lymphoproliferative disorders. Leukocytosis, splenomegaly, proliferation of T cells and virus production by lymphocytes characterized this infectious mononucleosis-like disease. Approximately two copies of circular herpesvirus sylvilagus genomes per cell were detected in spleen cells at 2 weeks postinfection, and circular genomes could still be observed after 4 months. Circular viral genomes were found in both B and T lymphocytes. Small amounts of linear viral DNA (0.1 to 0.3 copies per cell) were also detected in both B and T cells. These results indicated that the virus did not replicate in the majority of lymphocytes in vivo. Herpesvirus sylvilagus infection in cottontail rabbits could be useful as a model for studying the complex virus-host relationships of lymphotropic herpesviruses and perhaps as an animal model for Epstein-Barr virus infection in humans. PMID- 2993668 TI - Nucleotide sequence of a human cytomegalovirus DNA fragment encoding a 67 kilodalton phosphorylated viral protein. AB - Previously, we demonstrated that a DNA fragment from the L-unique region at 0.37 to 0.39 map units of human cytomegalovirus DNA codes for a 67-kilodalton phosphorylated tegument protein. (Davis et al., J. Virol. 52:129-135, 1984). We have determined the nucleotide sequence of this DNA, transcription initiation and termination, and putative promoter region. We also compared the properties of the protein product predicted from the DNA sequence with the properties of the protein isolated by immunoabsorbent column chromatography. This viral structural protein may have important functions, such as protein kinase activity, DNA binding, and possible transcriptional activation of immediate-early genes. PMID- 2993669 TI - Molecular cloning and nucleotide sequence of deer papillomavirus. AB - The genome of deer papillomavirus (DPV) isolated from American white-tailed deer was cloned into pBR322, and the entire nucleotide sequence of 8,374 base pairs was determined. The overall genetic organization of the DPV genome was similar to that of other papillomaviruses. All significant open reading frames were located on one strand, and the locations of putative promoters and polyadenylation signals were similar to those identified in the closely related bovine papillomavirus type 1 (BPV-1) genome. The DPV genome was approximately colinear with BPV-1 except for a noncoding region separating the early and late regions. The regions of highest nucleotide sequence homology between DPV and BPV-1 were found in the E1 open reading frame coding for BPV-1 DNA replication function and in the L1 open reading frame, which encodes the major capsid protein of BPV-1. PMID- 2993670 TI - Errant processing and structural alterations of genomes present in a varicella zoster virus vaccine. AB - Five minority populations of aberrant, varicella-zoster virus (VZV)-derived genomes were identified among the encapsidated DNAs obtained from the nuclear and cytoplasmic fractions of an in vitro infection initiated with a lyophilized sample of the BIKEN VZV vaccine (strain Oka). These were (i) VZV genomes, present within nuclear but not cytoplasmic viral capsids, which had been cleaved at a specific site within the short segment and which were, therefore, 3.15 megadaltons (approximately 4% of the VZV genome length) short of full length; (ii) highly deleted, repetitive VZV genomes which contained the errant cleavage site but not the usual VZV genome terminal sequences; (iii) VZV genomes into which multiples of 1 through 5 defective genome repeat units had been inserted into a homologous site; (iv) VZV genomes with additions of 0.1 or 0.18 megadaltons of DNA at both the terminal and internal ends of the short segment; and (v) VZV DNA which had lost the HindIII restriction site at map position 0.11. PMID- 2993671 TI - Cyclophosphamide, doxorubicin and cisplatin chemotherapy for patients with locally advanced urothelial tumors with or without nodal metastases. AB - Thirty-eight patients with primary urothelial tumors recurring in the pelvis or nodal presentation were treated with intravenous or intra-arterial cyclophosphamide, doxorubicin and cisplatin chemotherapy. The 38 patients were selected owing to unresectability by local criteria (12 patients) or by virtue of nodal metastases (26 patients). Histologically, the patients either had pure transitional cell carcinoma (29), transformation to a histological subtype of transitional cell carcinoma (7) or pure squamous cell carcinoma (2). An over-all 50 per cent complete remission rate was achieved with an 18 per cent objective pelvic response rate, and 32 per cent failed to respond to chemotherapy. Responses by histological subtype revealed that patients with pure transitional cell carcinoma had a 62 per cent complete remission rate, those with transitional forms had a 14 per cent complete remission rate and none with squamous cell carcinoma responded to chemotherapy. A significant difference in the incidence of responses among patients with transitional carcinoma and those with transition forms was seen (p less than 0.02). Complete remissions were independent of disease site. Nineteen patients achieved a complete remission with a mean duration of 86 weeks and median of 81 weeks (range 33 to 172 weeks). Toxicity of the chemotherapy was moderate with a high incidence of peripheral neuropathy and leukopenic infections. No deaths of chemotherapy were encountered. Patients with locally advanced or metastatic transitional cell carcinoma of the urothelium to lymph nodes can benefit from intravenous and intra-arterial cyclophosphamide, doxorubicin and cisplatin chemotherapy. PMID- 2993672 TI - Unilateral lymphadenectomy in intraoperative stage I nonseminomatous germinal testis cancer. AB - Bilateral retroperitoneal lymphadenectomy is mainly a staging procedure in patients with stage I nonseminomatous testis cancer, and it causes permanent loss of antegrade ejaculation in approximately two-thirds of the cases. Between May 1978 and August 1981, 61 consecutive patients with no intraoperative evidence of lymph node involvement underwent unilateral retroperitoneal lymph-adenectomy for nonseminomatous germinal testis tumors. Microscopic metastases were found in 1 to 4 retroperitoneal nodes in 6 cases (9.8 per cent). Antegrade ejaculation was absent postoperatively in 11 patients (18 per cent), with no significant difference between patients who underwent lymph node dissection on the left or right side. Ejaculation returned spontaneously in 3 patients, 1 of whom fathered a child. The disease recurred in 10 patients 3 to 35 months after lymphadenectomy (median 6 months). Disease recurred in 8 of 55 patients (14.5 per cent) with negative nodes and 2 of 6 (33.3 per cent) with positive histological findings. No patient suffered retroperitoneal recurrence. The more than 3-year survival rates free of disease were 96.4 and 83.3 per cent in patients with pathological stages I and II disease, respectively. Unilateral retroperitoneal lymphadenectomy in patients with intraoperative stage I nonseminomatous germinal testis cancer preserves antegrade ejaculation in more than 80 per cent of the cases without apparently compromising the long-term survival. PMID- 2993673 TI - Nonchromaffin paraganglioma of the retroperitoneum. AB - We reviewed 37 cases described previously in the literature to provide a patient operated upon for a retroperitoneal nonchromaffin paraganglioma with qualified advice concerning followup examinations and prognosis. This type of tumor apparently grows slowly and metastasizes late. A radical operation is the treatment of choice and provides a good prognosis. We suggested that our patient undergo clinical followup and ultrasound investigations at 3 to 6-month intervals. PMID- 2993674 TI - Testicular germ cell tumors after clomiphene therapy for subfertility. AB - The stimulatory effect of low doses of clomiphene citrate on the human male gonad is accepted widely. We used 25 mg. clomiphene daily for 3 weeks, with 1 week of rest, for 6 to 12 months in 650 oligospermic men without increased serum levels of follicle-stimulating hormone (less than 20 IU/l.). Unilateral testicular neoplasm developed in 2 subjects. Although a definite cause-and-effect relationship between treatment and tumor development cannot be assigned the possible risk of tumor development in men undergoing long-term clomiphene treatment is suggested. Since clomiphene administration or gonadotropin treatment might enhance tumor development in men, patients at risk undergoing such therapy should be examined frequently and when a testicle enlarges a malignant tumor must be ruled out. PMID- 2993675 TI - Experimental observations on dissolution of uric acid calculi. AB - An in vitro model was devised to evaluate the efficacy of the different irrigating solutions utilized for local dissolution of uric acid stones. Tris (hydroxymethyl) aminomethane proved to be several times faster than sodium bicarbonate in dissolving uric acid calculi. The maximal dissolution rate was obtained when the highest pH (10.5) of Tris buffer was used in concentrations at or above 0.2 M. This makes the commercially available THAM-E an optimal choice. Stones averaging 1 cm. in diameter were dissolved in less than 48 hours when this compound was used. Sodium bicarbonate should only be used in solutions with concentrations lower than 0.2 M and pH below 9, if some dissolution is to be attempted. Concentrations and pH's above these levels will coat the stones with hard shells of sodium urate, making it impossible to dissolve them. The in vitro findings were confirmed in vivo in a limited study in pigs with human uric acid calculi surgically placed in their kidneys. Our results indicate how to make the best use of the solutions clinically available in order to obtain total dissolution of uric acid stones in short periods of time. We recommend the use of a 0.3 molar concentration of this buffer (THAM-E) at flow rates of about 50 cc per hour. PMID- 2993677 TI - Femoral neuropathy after renal transplantation. AB - We report 2 cases of cadaveric renal transplantation in which the grafts were placed in the right iliac fossa. Postoperatively, both patients complained of ipsilateral thigh weakness. Electromyography and nerve conduction studies indicated femoral nerve neuropathy. Muscle weakness gradually resolved. Difficulty was encountered in placing both grafts, and each kidney and limb were subjected to prolonged ischemia. We suggest that the femoral neuropathy was ischemic in origin. PMID- 2993676 TI - Predictors of recurrence in stage II nonseminomatous testicular cancer after lymphadenectomy: implications for adjuvant chemotherapy. PMID- 2993678 TI - Testicular germ cell tumors in monozygotic twins: case report and review of the literature. AB - We report the eleventh instance of testicular germ cell tumors in monozygotic twins. The tumors were concomitant but of different histology. The comparable lymphography and computerized tomography scan findings, tumor response to chemotherapy and side effects are discussed. We conclude that there is no definite evidence for an increased risk in relatives of patients with this disease. PMID- 2993679 TI - Prognostic factors in stage III nonseminomatous testicular cancer. PMID- 2993680 TI - Viral infections of little blue penguins (Eudyptula minor) along the southern coast of Australia. AB - Serum antibodies to strains of avian paramyxovirus and flavivirus were detected in little blue penguins sampled at Port Campbell and Phillip Island, Victoria, Australia. No antibody to Newcastle disease virus (NDV) was detected in 267 sera collected, although one penguin captured for experimental studies had a hemagglutination-inhibition antibody titer of 2(4) to this virus. Experimental studies showed that the avian paramyxovirus designated APMV-IM and strain V4 of NDV were non-pathogenic for penguins, although the penguins could have been previously infected with these or similar virus strains. A flavivirus designated Saumarez reef virus, and an unnamed virus isolated from ticks on Macquarie Island, Southern Ocean were pathogenic causing disease and mortality in penguins inoculated with the viruses. PMID- 2993681 TI - Viral diseases of fish: first report of carp pox in golden ide (Leuciscus idus) in North America. AB - Carp pox, a putative viral disease exotic to North America, occurred in golden ide 1 yr after the fish were imported into the United States from the Federal Republic of Germany. The raised, white, plaque-like lesions, which occurred on about 5% of the fish, healed spontaneously and caused no mortality. Electron micrographs showed herpesvirus-like particles associated with lesion specimens; however, no infectious viruses were detected in tests with seven warmwater fish cell lines. PMID- 2993682 TI - Avian pox in a red-tailed hawk (Buteo jamaicensis). PMID- 2993683 TI - Leads from the MMWR. Recommendations for preventing possible HTLV-III/LAV virus from tears. PMID- 2993684 TI - HTLV-III antibodies in US Army blood donors in West Germany. PMID- 2993685 TI - Evidence for a 1980 HTLV-III infection in a currently asymptomatic B hemophiliac in Italy. PMID- 2993686 TI - Blood banks give HTLV-III test positive appraisal at five months. PMID- 2993688 TI - [Effects of anesthetics on the superoxide releasing activity of human polymorphonuclear leukocytes]. PMID- 2993687 TI - Effect of guar gum added to the diet of patients with duodenal ulcer. AB - In a randomized, double-blind, controlled clinical study, the effects of 5 g of guar gum, a dietary fiber composed of galactose and mannose, or placebo added to the diet of 20 patients with duodenal ulcer for 1 wk each were examined. Ten patients derived evident benefit and five some help from guar gum, on comparing symptoms during administration of guar gum with those experienced earlier or during the placebo week, whereas four patients found that neither guar gum nor placebo had any effect (p less than 0.001). The beneficial effect was associated with increased feelings of repletion after meals. Patients with fewest symptoms benefited only slightly, or not at all, from guar gum. In one patient, guar gum abolished pain felt earlier and on placebo, but also caused severe gastric retention after meals. This patient had pyloric stenosis. In patients who were intolerant to berries, fruits, sugar, sweet rolls, and pizza these foodstuffs were better tolerated during guar gum administration. The diarrhea which occurs in some patients ingesting guar gum was avoided by giving low initial doses. In three patients unpalatability of guar gum was a minor complaint. It is concluded that guar gum is helpful to many patients with uncomplicated duodenal ulcer, but that it is harmful to those having increased gastric emptying, eg, pyloric stenosis patients, and that guar gum may exert its effects by increasing gastric emptying time. PMID- 2993689 TI - [Effect of hypoxia on the content of endogenous opioid peptides and cyclic nucleotides in the rat brain]. PMID- 2993691 TI - [Clinical significance of CEA levels in lung cancer]. AB - Serum concentrations of CEA, Ferritin, beta 2-MG, TPA and IAP were measured in patients with lung cancer. Significant difference of positive rate between lung cancer and benign chest disease was observed only in CEA and TPA. CEA elevations occurred most frequently with stage IV and histologically with adenocarcinoma. Positive CEA levels (greater than or equal to 5 ng/ml) of pleural effusion were observed in 75% of all patients with lung cancer and increased to 94% in patients with positive cytology of effusion. There was not significant difference of bronchial washings CEA levels between lung cancer and benign chest disease. Furthermore, serial measurement of postoperative CEA levels had considerable value in monitoring the course of disease following resection. PMID- 2993690 TI - [Clinical assessment of sulbactam/cefoperazone in comparison with ceftizoxime in patients with postoperative infections by well controlled method]. AB - The clinical effectiveness in postoperative infections of sulbactam/cefoperazone (SBT/CPZ, (SBT 0.5 g+ CPZ 0.5 g) X 2/day) was compared to that of ceftizoxime (CZX, 1.0 g X 2/day) by a well controlled comparative study, to have the following results. The overall effectiveness rate of SBT/CPZ and CZX as judged by Judgement Committee was 84.0% (63/75) and 80.6% (50/62), respectively, and the effectiveness of SBT/CPZ and CZX as assessed by the attending surgeons was 84.0% (63/75) and 71.0% (44/62), respectively. No significant difference was noted in both assessments. In a total of 36 SBT/CPZ-treated patients with intraabdominal infections, the clinical efficacy was judged by attending surgeons to be excellent in 13 patients (36.1%), and to be excellent or good in 31 (86.1%). In the 30 CZX treated patients, it was judged to be excellent in 6 patients (20.0%), and to be excellent or good in 19 (63.3%). These results presented a significant difference (P less than 0.05, U-test) between the 2 drug groups. The final global improvement ratio judged by attending surgeons was 85.3% (64/75) for SBT/CPZ, and 79.0% (49/62) for CZX with no significant difference. In assessment of time course improvement, the improvement ratio of SBT/CPZ on day 4 was significantly better than that of CZX (P less than 0.05, U-test). The usefulness rate of SBT/CPZ and CZX was 84.0% (63/75) and 73.0% (46/63), respectively. There was no significant difference between the 2 drug groups. To assess the bacteriological efficacy, the eradication rate of SBT/CPZ was compared to that of CZX. There was no significant difference between 85.7% (36/42) for SBT/CPZ and 73.5% (25/34) for CZX. After SBT/CPZ administration, 2 patients (2.5%) complained of side effects. In the clinical laboratory tests, abnormality related to SBT/CPZ medication was observed in 6 patients (7.5%), and that related to CZX, in 5 patients (6.4%). As to the types of side effects and frequency, no significant difference was observed between SBT/CPZ and CZX. It is concluded from the above assessments that SBT/CPZ is a useful drug in the treatment of post-operative infections. PMID- 2993693 TI - [Photodynamic therapy in lung cancer]. AB - 242 patients with malignant tumors of various organs were treated by photodynamic therapy since January 1980. In this study the effects of photodynamic therapy for lung cancer was evaluated in 100 cases. According to the UICC clinical stage, 19 cases were stage Ia and were strongly suspected early stage on the basis of endoscopic findings, 16 were stage Ia and Ib, 11 were stage II, 38 were stage III and 16 were stage IV. This new therapy was shown to be effective in the curative treatment of early stage and in the palliative treatment of advanced lung cancer cases. Side effects included sensitivity to sunlight in 88%, dermatitis in 4%, bronchial perforation in 2% and obstructive pneumonia in 4%. PMID- 2993692 TI - [Studies on CA 125, CA 19-9, TPA and IAP as tumor markers in patients with ovarian cancer]. AB - CA 125, CA 19-9, TPA and IAP were useful for diagnosis of ovarian cancer. CA 125 was localized immunohistochemically in the surface and cytoplasma of cancer cells of serous cystadenocarcinomas, clear cell carcinomas, endometrioid carcinomas and undifferentiated carcinomas and CA 19-9 localized in mucinous cystadenocarcinomas and endometrioid carcinomas. When the cut off value of CA 125 was set by 35 U/ml in the control group, the patients with serous cystadenocarcinoma showed positive in 81%, resulting by 1,766.5 U/ml as an average value. As for CA 19-9 having set by 37 U/ml as the cut off value, patients suffering from mucinous cystadenocarcinoma who revealed positive by 60%, were 194 U/ml in an average value. It is considered that measuring values of CA 125 and CA 19-9 are valuable not only to diagnose as ovarian cancers but also to evaluate their prognosis. PMID- 2993695 TI - [Clinicopathological studies on disseminated carcinomatosis of the bone marrow occurring through metastasis of gastric carcinoma]. AB - Five cases of gastric carcinoma (one woman) were investigated. The average age of the patients was 44 and the clinical manifestation were lumbago, melena and fatigue. Two cases were Borrmann III, and other were Borrmann IV, IIc and IIc advanced, respectively. Three were signet-ring cell carcinoma and the other two were moderately differentiated and well-differentiated adenocarcinoma, respectively. All patients showed leukoerythroblastosis. Bone marrow specimens revealed myelofibrosis in three cases, while adenocarcinoma cells were also found in three. MHA or DIC was found in three patients, all of whom died of bleeding tendency. Mean survival time was 2.6 months. Thus, bone marrow carcinomatosis proved to be very severe. PMID- 2993694 TI - [Epidural spinal cord stimulation: its efficacy and mechanisms]. AB - Pain-alleviating effects of epidural spinal cord stimulation (ESCS) were assessed in patients with chronic intractable pain including cancer pain, and some aspects concerning its mechanisms were discussed. The temporary ESCS with percutaneously inserted electrodes was employed in 105 patients, and the implantable systems for long-term use in 19 patients. The ESCS had satisfactory effects especially in patients suffering from cancer pain, causalgia, facial and nape pain. The conditioning stimuli applied to the cervical dorsal cord exhibited interactions with the segmentally evoked spinal cord potential (SCP). The N1 wave of the SCP was inhibited up to 120 ms, while the P2 wave was facilitated for more than 100 ms, suggesting that the presynaptic inhibitory action at the dorsal horn is responsible for mechanism of the ESCS. CSF concentration of norepinephrine was significantly decreased by the ESCS therapy, indicating the existence of the relationship between norepinephrinergic descending inhibitory system and the ESCS in pain-alleviating mechanism. PMID- 2993697 TI - [Prognostic factors in adenocarcinoma of the colon and rectum]. AB - In this series of 986 patients with adenocarcinoma of the colon and rectum, the value of different prognostic factor was discussed. The five-year survival rate was 60.1% overall. The patient's sex and age and the location of the carcinoma were not correlated with survival. The protuberant type macroscopically, less than 2.9 cm in diameter, under one third in circumference and well-differentiated adenocarcinoma were relatively good prognostic factors, but the majority of these cases had adenocarcinoma confined to the bowel wall. The depth of invasion, nodal metastasis and venous invasion were significant indicators of survival. PMID- 2993696 TI - [Evaluation of the malignancy of breast carcinoma using flow cytometry]. AB - Flow cytometric DNA analyses of single cell nuclei were performed on nuclear suspensions prepared from tumors of primary breast carcinomas in 40 patients operated on in our hospital. Fluorescence intensities of propidium-iodide stained cell nuclei were analyzed in a SHOWA DENKO SC-20 cell sorter. The DNA index and the percentage of S-phase cells and DNA aneuploidy were calculated from each DNA histogram. There was a tendency for more advanced nodal tumor involvement to have a higher DNA index, increasing DNA aneuploidy proportions and significantly larger percent of S-phase cells. PMID- 2993698 TI - [The effect of leukotriene B4 (LTB4) on chemotaxis of human peripheral blood monocytes]. PMID- 2993700 TI - [Biochemistry of neurotransmitter receptors]. PMID- 2993699 TI - [Membrane lipids and membrane enzymes]. PMID- 2993701 TI - [Theoretical classification of hormone receptor abnormalities, with special reference to diabetes mellitus]. PMID- 2993702 TI - [Hereditary metabolic diseases caused by membrane abnormality]. PMID- 2993703 TI - [Drug resistance and withdrawal syndrome--study on receptors]. PMID- 2993704 TI - [Natriuresis--the endogenous digoxin-like-factor and atrial natriuretic polypeptide]. PMID- 2993705 TI - [Catecholamine receptors and antagonists: b. Presynaptic receptors]. PMID- 2993706 TI - [Angiotensin converting enzyme inhibitors]. PMID- 2993707 TI - [Pharmacology of the kallikrein-kinin system]. PMID- 2993708 TI - [A case of islet cell tumor]. PMID- 2993709 TI - [Calcium-activated neutral protease in the porcine skin]. PMID- 2993710 TI - [Immunohistological identification of papillomavirus antigen in cutaneous warts using anti-papillomavirus serum]. PMID- 2993711 TI - [An eosinophil chemotactic factor of malignant fibrous histiocytoma]. PMID- 2993712 TI - Phosphotyrosine phosphatase activity in human placenta. AB - A phosphotyrosine phosphatase was enriched 9-fold from human placenta homogenates using ammonium sulfate fractionation and chromatographies on P-cellulose and Phenyl Sepharose. The enzyme preparation hydrolyzed phosphotyrosine, p nitrophenyl phosphate, as well as phosphoenol-pyruvate. The enzyme was unstable, and its Km value for phosphotyrosine increased during the purification procedure. The optimum pH for phosphotyrosine was found to be pH 7.0. The phosphotyrosine phosphatase activity was completely inhibited at 2.6 mM Cu2+. Zn2+ was slightly inhibitory to the phosphatase activity. The effects of several inhibitors on the phosphotyrosine phosphatase activity were different from those on the p nitrophenyl phosphatase activity. These results suggest that the phosphotyrosine phosphatase activity from human placenta is different from previously reported acid and alkaline phosphatases. PMID- 2993713 TI - Isozymes of human erythrocyte pyrimidine 5'-nucleotidase--chromatographic separation and their properties. AB - The biochemical properties of two isozymes of human erythrocyte pyrimidine 5' nucleotidase (P5N) separated by ion-exchange chromatography were studied. These two isozymes, P5N-I and P5N-IIB, can be distinguished from each other by substrate specificity, pH dependent activity and thermostability. P5N-I is thermolabile and dephosphorylates various pyrimidine 5'-monophosphates optimally around pH 7. UMP and CMP were found to be most effective substrates. P5N-IIB is thermostable and characterized by its high Michaelis constant and maximum velocity for dUMP or dTMP and acidic pH optima. The molecular weight was estimated to be 39,000 for P5N-I and 44,000 for P5N-IIB by gel filtration. These data suggest that these two isozymes are independent proteins and probably encoded by distinct structural gene loci. PMID- 2993714 TI - [Observation of both intra- and extra-hepatic vessels by real time two dimensional Doppler echography--its clinical significance and prosthesis]. PMID- 2993715 TI - [Study of 45 patients with hepatocellular carcinoma complicating occlusion of the main portal vein]. PMID- 2993716 TI - [Cancer detection with 18F-5-fluorodeoxyuridine--a new cancer diagnostic agent reflecting nucleic acid metabolism]. PMID- 2993717 TI - Binding characteristics of 3H-dihydroalprenolol to beta-adrenergic receptors of rat brain: influence of lectins. AB - The significance of the carbohydrate moieties of the beta-adrenergic receptor molecule in the rat brain was examined using the radioligand binding assay method. Thus, this experiment was designed to assess the effects of lectins, concanavalin A (Con A), Phaseolus vulgaris agglutinin (PHA), and wheat germ agglutinin (WGA) on the affinity of the beta-adrenoceptor. The rat brain was used and the beta-adrenoceptor binding assay was carried out using 3H dihydroalprenolol as a ligand. Con A and PHA significantly caused an increase in the values of the density of beta-adrenoceptor (Bmax) and a reduction in the values of the dissociation constant (Kd), but significant changes were not observed with WGA. These results strongly suggest that the carbohydrate moieties of the cell surface containing the beta-adrenoceptor molecule may have a crucial role in the drug-receptor interaction, and they imply that the beta-adrenoceptor molecule is a glycoprotein which contains N-linked carbohydrate chains. PMID- 2993718 TI - Mechanism of relaxant action of papaverine. VI. Sodium ion dependence of its effect on 45Ca-efflux in guinea-pig taenia coli. AB - The present study was undertaken to investigate the roles of sodium ion and the cyclic AMP system in the relaxant effect of papaverine. The effects of papaverine on the 45Ca-efflux and the mechanical activity of guinea-pig taenia coli were tested in solutions in which the concentration of sodium ion was varied and compared with those of dibutyryl cyclic AMP. Papaverine dose-dependently caused an acceleration of 45Ca-efflux and the synchronous relaxation of a depolarized preparation in normal bathing solution. Dibutyryl cyclic AMP mimicked these effects of papaverine. In Na-free solution, papaverine lost its ability to accelerate the 45Ca-efflux, and its relaxant activity was markedly reduced, while dibutyryl cyclic AMP had neither an effect on the 45Ca-efflux nor an effect on the muscle relaxation in Na-free solution. Reintroduction of a small amount of sodium ion to the solution, however, recovered these effects of papaverine and dibutyryl cyclic AMP on the 45Ca-efflux and the muscle relaxation. These findings indicate that the relaxant effect of papaverine may be in part due to an increase in cyclic AMP-mediated Ca-efflux which requires the presence of external sodium ion. The sodium ion dependence of this Ca-efflux process was also discussed. PMID- 2993719 TI - Sources of extramitochondrial corticoidogenic cholesterol in the adrenal cortex. AB - Intracellular sources of extramitochondrial corticoidogenic cholesterol in bovine, rat and hamster adrenocortical cells were examined in vitro by comparing the species differences in the effects of various inhibitors on the adrenocorticotropic hormone (ACTH)-induced corticoidogenesis. The inhibitors were ML-236B (3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor), W 7 (N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide; calmodulin inhibitor), dichlorvos (O,O-dimethyl-2,2-dichlorovinyl phosphate; organic phosphorylation inhibitor), chloroquine [7-chloro-4-4-diethylamino-1-methyl-butylamino) quinoline; lysosomal enzyme inhibitor) and cycloheximide (protein synthesis inhibitor). During 2 to 3 hr incubation periods, the ACTH-induced corticoidogenesis was not inhibited by ML-236B (100 microM) in the bovine and rat adrenocortical cells. In the hamster adrenocortical cells, ML-236B (100 microM) did not affect the ACTH-induced corticoidogenesis during the initial 1 hr incubation periods; but thereafter, the ACTH-induced corticoidogenesis during the subsequent 2 hr incubation periods was completely blocked by ML-236B. The ACTH induced corticoidogenesis was inhibited by W-7 (up to 25 microM) in the bovine and rat adrenocortical cells, but this was not the case in the hamster cells. Chloroquine (up to 400 microM) inhibited the ACTH-induced corticoidogenesis in the adrenocortical cells of three different species, but the hamster adrenal cells were much more vulnerable than the bovine and rat cells. The ACTH-induced corticoidogenesis in the adrenocortical cells of three different species were equally inhibited by cycloheximide (up to 1 mM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993720 TI - Effects of angiotensin I-converting enzyme inhibitor (SA-446) on renal function in dogs. AB - The effects of an orally active inhibitor of angiotensin-converting enzyme (SA 446) on systemic arterial pressure, renal function and renin release were examined in anesthetized dogs. Intrarenal infusion of the larger dose of SA-446 (0.1 mg/min) caused an increase in RBF, urine flow and renin release and caused a fall in blood pressure. The smaller dose of SA-446 (0.02 mg/min) did not affect the blood pressure, but it increased the urine flow. However, the same dose of SA 446 in combination with probenecid caused a significant fall in blood pressure. The potentiation of SA-446 with probenecid may be explained by the elevation in plasma SA-446 concentration via the inhibition of its tubular secretion by probenecid. PMID- 2993721 TI - [The metabolism of trace metals in patients with lung cancer--with special reference to zinc]. PMID- 2993722 TI - [A case of iliac arterial embolism after lung cancer resection]. PMID- 2993723 TI - Production and characterization of monoclonal antibodies to mouse hepatitis virus, MHV-NuU. PMID- 2993724 TI - Role of T cells in feline infectious peritonitis virus infection of suckling mice. PMID- 2993725 TI - Transfer of antibodies against viruses of calf diarrhea from cows to their offspring via colostrum. PMID- 2993726 TI - A case of neonatal calf diarrhea associated with natural infection with rotavirus. PMID- 2993727 TI - Parvoviral myocarditis in cyclophosphamide-treated chicks. PMID- 2993728 TI - Diversity of melanoma plasma membrane proteinases: inhibition of collagenolytic and cytolytic activities by minocycline. AB - The tissue-destructive proteinases of B16-BL6 melanoma cells from C57BL/6 mice and subcellular fractions were examined. Cancer cell organelles were isolated following nitrogen cavitation with the use of sucrose density gradient centrifugation. Serine, cysteine, and metalloproteinases were assayed with the use of radiolabeled proteins and synthetic substrates. Tumor-induced red blood cell lysis was quantitated by measurement of the release of isotope from 59Fe labeled red blood cells (RBC) cocultivated with melanoma cells; the RBC were from Wistar rats. Enzyme inhibitors with specificity toward different classes of proteinases were used in the above assays to categorize the enzymes responsible for substrate degradation. Results indicated that intact melanoma cells, cell organelles, and cytosol contain proteinases that can degrade collagen and gelatin and lyse normal RBC. Melanoma plasma membranes are highly enriched in collagenase, gelatinase, cysteine proteinase, plasminogen activator, and cytolytic activity. The inhibition of tumor collagenolytic, gelatinolytic, and cytolytic activities by EDTA and 1,10-phenanthroline but not by diisopropyl fluorophosphate and N alpha-p-tosyl-L-lysine chloromethyl ketone indicates that metalloproteinases are the active enzymes in these assays. Minocycline, a synthetic tetracycline with demonstrable inhibitory activity with other mammalian collagenases, also inhibited melanoma collagenolytic and cytolytic activities. PMID- 2993730 TI - Effects of various dietary staples on esophageal carcinogenesis induced in rats by subcutaneously administered N-nitrosomethylbenzylamine. AB - Previous epidemiologic studies associated large differences of esophageal cancer risk with the nature of the staple diet. In this study, various cereals and dietary staples were fed to inbred BD IX rats for 7 months or longer. N Nitrosomethylbenzylamine [(MBN) CAS:937-40-6] was given five times subcutaneously between the 45th and 58th day. The percentage of rats with tumors and the mean number of tumors per esophagus were similar when corn, wheat, commercial bird resistant sorghum, bananas, and polished rice were fed but were strikingly lower when the basis of the diets was millet, red sorghum, brown rice, or potatoes. The number of esophageal tumors was significantly related to the dietary concentration of some minerals and vitamins. Supplementing marginally deficient corn or wheat diets with various combinations of nicotinic acid, riboflavin, zinc, magnesium, molybdenum, and selenium significantly reduced the numbers of esophageal tumors. When the feeding of protective cereals or nutrients was commenced only 150 days after MBN was given, a marked inhibitory effect on the progression of tumors was still observed. PMID- 2993731 TI - Histochemistry of liver tumors induced by diethylnitrosamine and differential sex susceptibility to carcinogenesis in Oryzias latipes. AB - Liver tumor nodules were induced by the administration of diethylnitrosamine [(DENA) CAS: 55-18-5; N-nitrosodiethylamine] to the fish, Oryzias latipes. A histochemical study showed decreased ATPase and glucose 6-phosphatase (Glc-6 Pase) activities in most of the tumor nodules. Increased ATPase staining and, occasionally, Glc-6-Pase staining were also observed. In some nodules, the distribution of bile canaliculi was disordered, indicating positive ATPase activity. Basophilic and eosinophilic nodules could be discriminated by histologic examination. The observations on serial histochemical and histologic sections revealed extreme heterogeneity in the phenotypes of the nodules. Measurement of the enzyme-altered areas indicated that the development of nodules was more prominent in male than in female fish. Experiments with sex-reversed fish, XX males and XY females, suggested that the sex difference in the susceptibility to DENA does not result from the difference in sex chromosomes but from the difference in sexual phenotypes. PMID- 2993729 TI - Ultrastructural and cytochemical study of the early stages of liver colonization by transplanted neoplastic hepatocytes. AB - Neoplastic liver cell colonies were induced in the livers of isogeneic F344 rats by intraportal injection of a hepatic cell suspension from diethylnitrosamine treated donor rats. Examination of the livers 12 days after cell implantation revealed well-demarcated groups of liver cells. The colonies showed alterations of the normal hepatocyte phenotype, which were clearly demonstrated by histologic, cytochemical, and electron microscope techniques. The hepatocytes were markedly deficient in glucose-6-phosphatase and bile canalicular ATPase activities, and they contained numerous mitotic figures. Scanning and transmission electron microscopy allowed characterization of hepatocyte interfaces and the shape of sinusoids and the biliary network. The nodular colonies displayed disorganized, thickened trabeculae separated by dilated sinusoids. In these colonies the hepatocytes proliferated intensely and formed, inside the host parenchyma, revascularized, integrated nodular structures. However, these hepatocytes showed ultrastructural anomalies: large nuclei with prominent nucleoli, many free polysomes, and areas of proliferated smooth endoplasmic reticulum in connection with unfolded cisternae of the rough endoplasmic reticulum. All of these features agreed with the hypothesis previously proposed that the colonies may be precursors of the hepatocarcinomas that ultimately develop in animals given injections of treated liver cells. Direct confirmation, however, still is needed. PMID- 2993732 TI - [Treatment of patients with disseminated malignant testicular tumors using the Bulgarian preparation biocisplatinum and its administration regimens]. PMID- 2993733 TI - The effect of wheat bran on the pharmacokinetics of phenprocoumon in normal volunteers. AB - Observations of a variable anticoagulatory response in patients being treated by phenprocoumon and wheat bran resulted in a study of seven healthy volunteers. Up to 168 h after the administration of phenprocoumon, both in the fasting state and following the ingestion of 35 g wheat bran in a crossover design, blood samples were taken and urine was collected. The usual pharmacokinetic parameters were calculated from the concentrations measured. Following the ingestion of wheat bran, a decreased absorption rate for phenprocoumon but no decrease in overall bioavailability was observed. In addition, a decrease in total body clearance of phenprocoumon and an increase in the free plasma fraction of phenprocoumon, resulting in decreased free drug clearance, was seen after wheat bran administration. No differences were observed in the urinary excretion of either phenprocoumon or its metabolite. These findings cannot be completely explained by our present knowledge and need further investigation. PMID- 2993736 TI - [Case of a giant androsteroma]. PMID- 2993734 TI - Lymphadenopathy and antibodies to HTLV-III in homosexual men. Clinical, laboratory and epidemiological features. AB - The study provides information on the epidemiology of HTLV-III infection and the lymphadenopathy syndrome (LAP) in 374 German homosexual men. Sexual contacts in the USA and rectal enemas before receptive anal intercourse are the main risk factors associated with virus transmission. HTLV-III seropositivity is significantly correlated with LAP. Prominent clinical signs are infrequent. Immunological and haematological abnormalities are prevalent, and the retrovirus infection is frequently associated with serological markers of other viruses (hepatitis B, herpes group viruses). Lymphadenopathy as a manifestation of HTLV III infection is discussed within the context of AIDS-related disorders. PMID- 2993735 TI - Ketoconazole blocks cortisol secretion in man by inhibition of adrenal 11 beta hydroxylase. AB - We investigated basal and ACTH stimulated levels of cortisol, corticosterone, 17 alpha-hydroxyprogesterone, 11-deoxycortisol and 11-deoxycorticosterone as well as plasma levels of ACTH before and during the oral administration of ketoconazole in five patients with Cushing's syndrome (3 with bilateral adrenal hyperplasia, 1 with adrenal adenoma and 1 with adrenal carcinoma) and in three controls. The influence of ketoconazole on the transformation of 3H-17 alpha hydroxyprogesterone to 3H-11-deoxycortisol and 3H-cortisol and of 3H-11 deoxycortisol to 3H-cortisol as well as of 3H-11-deoxycorticosterone to 3H corticosterone was also examined in slices or homogenates of normal and hyperplastic adrenal tissue from four patients. Ketoconazole induced a rise of 11 deoxycortisol and 11-deoxycorticosterone, but not of cortisol and inconsistently of corticosterone which were increased by ACTH. Thus the ratio 11 deoxycortisol/cortisol rose more after ketoconazole than after ACTH and the ratio 11-deoxycorticosterone/corticosterone rose after ketoconazole but fell after ACTH. Plasma ACTH levels were stimulated 2-50 fold by ketoconazole. Incubation studies of adrenal tissue slices with 3H-17 alpha-hydroxyprogesterone showed that ketoconazole inhibited the transformation of 3H-17 alpha-hydroxyprogesterone to 3H-cortisol but not to 3H-11-deoxycortisol so that the ratio 3H-11 deoxycortisol/3H-cortisol increased 15-80 fold. After incubation of adrenal slices with 3H-11-deoxycortisol or 3H-11-deoxycorticosterone and ketoconazole, a 2-260 fold increase of the ratios 3H-11-deoxycortisol/3H-cortisol and 3H-11 deoxycorticosterone/3H-corticosterone were also found. PMID- 2993737 TI - [Protein kinase activity in the gastric mucosa of patients with chronic gastritis and stomach tumors]. PMID- 2993738 TI - [A rare case of Wilms' tumor in an adult]. PMID- 2993739 TI - A Yaba-like condition in a young baboon (Papio anubis). AB - A rapidly developing proliferation of subcutaneous nodules on the leg of a young baboon is described. Histologically the nodules consisted of plump round cells, many of which contained intracytoplasmic eosinophilic viral-like inclusions. PMID- 2993740 TI - Palatability and digestibility of soya- and milk proteins in Callithricidae. AB - 4 protein sources were tested for palatability, digestibility, influence on bodyweight and consistency of faeces in Callithrix jacchus and Saguinus fuscicollis. The protein sources were incorporated at crude protein levels of 12, 17 and 22% in a pelleted diet. A banana supplement was also fed which reduced the total protein content to 10.7, 14.7 and 18.3%. The test diets, fed for 3 weeks, were well accepted with the milk proteins being slightly preferred. Consumption of the 12% protein pellets was greater than their 22% equivalents. True protein digestibility was calculated to be 97% for lactalbumin, 96% for casein, 90% for soy protein concentrate and 83% for soyabean meal. Changes in bodyweight were negligible. Dry matter of faeces varied between 25% (lactalbumin diets) and 28% (casein diets), but did not change with increasing protein intake. Increasing levels of cellulose, from 2.4 to 7.2% total crude fibre lowered the apparent protein digestibility from 84.3 to 79.2% and the digestibility of dry matter from 82.2 to 76.5%. PMID- 2993742 TI - Lysosomal storage diseases. PMID- 2993741 TI - Role of canalicular contraction in bile flow. PMID- 2993745 TI - Pulmonary tuberculosis with peripheral neuropathy: the importance of a lateral chest film. PMID- 2993746 TI - Activation of microsomal cAMP-dependent protein kinase isoenzyme I by ACTH1-24 in bovine adrenal cells. AB - In microsomes of bovine fasciculata reticularis cells incubated with or without 10(-8) M ACTH during 20 min, we measured covalent and non covalent cAMP binding under exchange or non-exchange conditions and cAMP-kinase activity. ACTH induced a decrease in cAMP-kinase activity and in the number of free cAMP binding sites. These results indicate an activation by ACTH of a part of microsomal cAMP dependent protein kinase. Photoaffinity labeling of microsomal protein with 8 azido-cAMP revealed the presence of both cAMP-kinase isoenzyme I and II in this cellular fraction. Using this method, it was demonstrated that ACTH1-24 caused a preferential and nearly complete activation of microsomal protein kinase I. PMID- 2993744 TI - Analysis of the major metabolite of delta 9-tetrahydrocannabinol in urine. VI. Specificity of the assay with respect to indole carboxylic acids. PMID- 2993747 TI - Effect of ACTH and prolactin on dehydroepiandrosterone, its sulfate ester and cortisol production by normal and tumorous human adrenocortical cells. AB - The effect of ACTH and prolactin on the synthesis of dehydroepiandrosterone (DHEA) and its sulfate ester (DHEAS) was studied in cell suspensions of "normal" and tumorous (adenoma) human adrenal cortex. A stimulation of DHEA and no response of DHEAS production by ACTH in "normal" adrenocortical cell suspension was observed. However ACTH stimulated both DHEA and DHEAS synthesis in tumorous adrenocortical cells. Prolactin did not influence either the basal or the ACTH stimulated DHEA and DHEAS production of adrenocortical cells irrespective of their origin. Our results are compatible with the concept that the biosynthesis of DHEA is under ACTH control, while other factor(s) regulate(s) the sulfate pathway of DHEA secretion under normal conditions. In tumorous adrenocortical cells DHEA may be regulated--at least partly--by ACTH. Prolactin seems to have no direct effect on DHEA and DHEAS synthesis. It is postulated that the relationship between serum prolactin and DHEAS (or DHEA) levels observed by several authors might be an extraadrenal effect of prolactin on adrenal androgens. PMID- 2993748 TI - Effects of several pro-opiomelanocortin derived peptides on steroidogenesis in ovine and bovine adrenal cells. AB - The effects of several peptides derived from the amino-terminal end of proopiomelanocortin (N-POMC) alone or in combination with ACTH on ovine and bovine adrenal cell steroidogenesis have been studied. Neither porcine N-POMC1-61 amide, nor gamma 3-MSH, nor been studied. Neither porcine N-POMC1-61 amide, nor gamma 3-MSH, nor Lys-gamma 3-MSH alone had any steroidogenic effect while porcine N-POMC1-80 alone had a week but significant steroidogenic effect on isolated adrenal, the effect being more pronounced on cultural adrenal cells. The potentiation by N-POMC peptides of the steroidogenic action of ACTH was investigated using both freshly isolated and cultured adrenal cells. At 10(-8) M N-POMC1-61 amide, gamma 3-MSH and Lys-gamma 3-MSH did not modify the ACTH dose response for steroids (gluco- and mineralocorticoids) and cAMP production. Likewise, the stimulatory effect of 10(-12) M ACTH was not modified by increasing concentrations (10(-11) to 10(-8) M) of N-POMC1-61 amide or gamma 3-MSH. On the other hand, an additive effect of 10(-8) M N-POMC1-80 on the steroidogenic action of low concentration of ACTH was observed. This again was more pronounced in cultured adrenal cells. The same effects were observed when increasing concentrations of this peptide (10(-9) and 10(-8) M) were added together with 10( 12) M ACTH. This result indicates that the potentiating effects of N-POMC derived peptides on the steroidogenic effect of ACTH which has been described on rat and human adrenal, is not a general phenomenon in mammals. PMID- 2993743 TI - Surface charge distribution in normal and transformed rat bladder epithelial cells in vitro. AB - Cationized ferritin (CF) was used as an ultrastructural marker to study differences in the distribution and density of surface anionic charges between normal and neoplastic cells. In anchorage-independent cell systems, CF induces a redistribution of cell surface receptor-ligand complexes into clusters, patches, and caps on the surfaces of transformed cells, but not on the surfaces of normal cells. In the present report, the authors have, for the first time, extended the CF labeling studies to a system of anchorage-dependent, rat bladder epithelial cell lines: normal RBTC cells and RBTCC-8 carcinoma cells. Cells were grown to confluency and labeled with 500 micrograms/ml CF for 3 minutes. After completion of CF labeling, cells were incubated for 0, 15, 60, 120 minutes, or overnight and were fixed then with buffered glutaraldehyde for electron microscopy. The results show that (a) CF induces grouping of surface anionic charge sites into patches and clusters on RBTCC-8 carcinoma cells (CF covers 53.86 +/- 2.15% of the plasma membrane), but not on normal RBTC cells (CF covers the entire plasma membrane); (b) CF densities are well correlated to the cell surface sialic acid contents of RBTC (2.42 +/- 0.30 microM/10(9) cells) and RBTCC-8 cells (1.26 +/- 0.25 microM/10(9) cells); and (c) CF-labeled membrane is internalized at equal rates by RBTC and RBTCC-8 cells (30% in 15 minutes; 60% in 60 minutes; 70% in 120 minutes; 100% overnight). These data indicate that CF-induced patching of anionic sites is a surface characteristic that is common to anchorage-dependent and independent tumor cells. Mechanisms of CF patching in neoplastic cells are discussed. PMID- 2993749 TI - Reinforcement expectancies and alcoholism treatment outcome after a one-year follow-up. AB - The relationship of alcohol-related expectancies with the drinking behavior and treatment compliance of alcoholics was examined 1 year posttreatment, along with the utility of reinforcement expectancy as a predictor of alcoholism treatment outcome. Of the original sample of 42 men veteran inpatients from an alcoholism treatment program, 81% were interviewed 1 year after treatment. Prediction variables identified in the multiple-regression analyses accounted for 40-57% of the criterion variance. It was found that more limited expectancies of alcohol produced relaxation were associated with abstinence, as well as nonproblematic drinking. It was also found that there was a consistent negative linear relationship between alcohol expectancies and measures of treatment success. PMID- 2993750 TI - Comparison of three outpatient treatment interventions: a twelve-month follow-up of men alcoholics. AB - Men inpatient alcoholics (N = 174) from a Veterans Administration medical center who were preselected by employment status were randomly assigned to one of three outpatient treatment interventions: (1) medication only, (2) active support or (3) untreated medical monitoring. Subjects were followed monthly for 1 year, with an 85% 12-month follow-up rate. Although the sample as a whole showed reduced alcohol misuse and improved social functioning after 12 months, the specific form of treatment was unrelated to outcome. These findings suggest that the intensity of the outpatient treatment experience is not related to outcome and that time consuming interventions are not differentially cost-effective. PMID- 2993751 TI - [The spinal enkephalinergic and serotoninergic systems in the control of transmission of nociceptive messages]. AB - Numerous anatomical, pharmacological and electrophysiological data described in the literature indicate that spinal enkephalinergic and serotoninergic systems are probably involved in the control of nociceptive inputs from the periphery to the cerebral cortex. However, reported evidence was generally indirect and did not provide a real demonstration of the physiological participation of these neurones in pain control. This led us to select appropriate experimental approaches for studying directly the activity of spinal enkephalinergic and serotoninergic systems in animals (rat, cat) submitted to noxious stimuli. Owing to two catheters introduced into the subarachnoidal space of anesthetized rats, it was possible to perfuse the whole spinal cord with an artificial cerebro spinal fluid and thus collect the neuroactive compounds released by spinal neurones (at least those in superficial layers) under various experimental conditions. Using this technique, we observed that some (but not all) nociceptive stimuli such as intense pinching of the muzzle, intraperitoneal injection of acetic acid or noxious heat applied to the muzzle or the tail induced a significant increase in met-enkephalin release from the spinal cord (see fig. 2). Similar effects were observed following the blockade of enkephalin catabolism by thiorphan and bestatin (see fig. 1) indicating that they were not due to some alteration of peptidase activities but really involved the activation of spinal enkephalinergic systems. Since cervical cord transection suppressed the stimulatory action of noxious stimuli on spinal met-enkephalin release, it could be proposed that the mechanisms involved were not limited to the cord but depended on supraspinal structures. Bulbo-mesencephalic serotoninergic neurones projecting to the spinal cord might well correspond to such structures (or at least to some of them) since nociceptive stimuli (such as noxious heat applied to the tail) also evoked a marked increase of serotonin (5-HT) release at the spinal level (fig. 3). Such observations together with indirect evidence reported in the literature suggested therefore that the activation of spinal enkephalinergic systems triggered by noxious stimuli might result from excitatory influence due to descending serotoninergic projections. However, in vitro studies using slices of the dorsal zone of the rat lumbar cord did not reveal any stimulatory effect of 5-HT on the spontaneous or K+-evoked release of met-enkephalin (fig. 4).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993752 TI - [Enkephalinase inhibitors and molecular study of the differences between active sites of enkephalinase and angiotensin-converting enzyme]. AB - With the aim of producing an analgesia physiologically induced by endogenous opioids, several series of inhibitors of the degradation enzymes of enkephalins have been synthetized by using as a model, at the atomic level, the active site of thermolysin, a bacterial endopeptidase similar to enkephalinase. Thiorphan and retro-thiorphan are very potent inhibitors of enkephalinase (KI = 2 nM), but the retro compound is more selective, as it is unable to recognise the angiotensin conversion enzyme. Recently, a series of inhibitors containing a bidentate group were found to be capable of inhibiting the three metallopeptidases which break down the enkephalins. One of these compounds, kelatorphan, totally protects, in vitro and in vivo, Met-enkephalin from enzymatic degradation. Kelatorphan is the first complete inhibitor of enkephalin metabolism and is the only compound to possess an analgesic activity greater than that of a mixture of thiorphan and bestatin (non-specific aminopeptidase inhibitor). A tritiated derivative of kelatorphan has been used to visualise the enkephalinase in the rat brain by means of autoradiography. The enzyme has a heterogeneous distribution with a particularly high concentration in the nigro-striatal system. PMID- 2993753 TI - [Adrenergic and purinergic receptors and bronchial motoricity]. AB - Numerous studies have been, and are being devoted to the nature of adrenergic and purinergic receptors in the bronchopulmonary system. Studies of beta adrenoceptors performed with ligands (table I) have demonstrated the presence of two types of receptors, beta 1 and beta 2 in proportions of about 15 : 100 respectively; this proportion is approximately the same at all levels of the tracheobronchial tree. Beta-adrenoceptors (beta 1 + beta 2) are globally more numerous in peripheral organs which contain heterogeneous tissues. Their number can be modified in certain circumstances, notably in asthma, infection and after prolonged treatment with sympathomimetic amines. Functional studies using specific beta 1-adrenoceptor agonists (RO-363 or prenalterol) or determining the relative activities of beta 1 and/or beta 2 stimulants and their inhibition by selective beta-blockers have shown that stimulation of beta 1-adrenoceptors may produce partial relaxation of the isolated trachea but not of lung parenchyma, the latter being supposed to represent distal airways. Studies on isolated small bronchi, about 0.1 mm in diameter (fig. 1 and 2A) have confirmed that stimulation of beta 1-adrenoceptors has not effect on distal airways. They have also demonstrated that beta 2-stimulants have different intrinsic activities (fig. 2B). Studies of alpha-adrenergic receptors using ligands (table II) have shown that these receptors are in small number in the tracheobronchial tree of numerous animal species. Functional studies on the conscious guinea-pig have shown that clonidine can potentiate the bronchoconstrictor effects of acetylcholine, histamine and serotonin (fig. 3) and that this potentiating effect is specifically inhibited by yohimbine and piperoxan (fig. 4). This action of clonidine has been attributed to depression of the reflex sympathetic activity associated with bronchospasm. Alpha 1-adrenoceptor agonists (phenylephrine, methoxamine) induce contracture of the isolated bronchial smooth muscle (fig. 5) but may partially reduce the bronchoconstrictor effects of acetylcholine, histamine or serotonin (fig. 6). This last effect is partially inhibited by alpha 1-blockers (fig. 7) and seems to be due to shrinkage of the bronchial mucosa. Finally, studies of purinergic receptors in the bronchopulmonary system have shown that they probably are of the A2-P1 type (tables III and IV) and that they do not seem to be involved in the bronchodilator activity of theophylline. PMID- 2993754 TI - HPLC-ECD procedure for the measurement of O-methylation of catechol estrogens by vascular tissue. AB - A precolumn extraction procedure combined with high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed for the identification and quantitation of catechol estrogens, their immediate precursors (estrogens), and their 0-methylated metabolites. These compounds were isolated from Krebs solutions and from perchloric acid tissue extracts or whole tissues by simple solvent extraction prior to quantitation. The optimal conditions for detection by HPLC-ECD were those employing a radial compression module, a C18 Bondapak cartridge, and an isocratic mobile phase of ammonium phosphate (pH 3.0) combined with 30-70% acetonitrile. A biological application of this procedure includes the investigation of the 0-methylation of catechol estrogens by vascular tissue. In the present study, the 0-methylation of 2 hydroxyestradiol by rabbit aorta has been examined. The 0-methylation in segments of thoracic aorta was significantly less than that in abdominal aortic segments, but was reproducible along the length of the thoracic aorta. PMID- 2993755 TI - Non-local thermodynamic effects and efficiency of oxidative phosphorylation. AB - A non-equilibrium thermodynamic model of oxidative phosphorylation is formulated, which allows us to take into account some non-local effects. In this way, we compute the influence of the tangential resistivity of the inner mitochondrial membrane to proton current, as well as that of the distance between active sites, on the stoichiometry and efficiency of energy conversion. PMID- 2993756 TI - Electron microscopy in selection of patients with small cell carcinoma of the lung for medical versus surgical therapy. AB - Fifty-one cases of small cell carcinoma of the lung were studied by electron microscopy in order to determine if ultrastructural subsets could be found and if these subsets predicted clinical behavior. All of these cases were considered bona fide small cell carcinoma of the lung by light microscopy. Tumors with ultrastructural features of epithelial differentiation were defined by the presence of well-formed, classic desmosomes joining adjacent cells and by additional features of squamous or glandular differentiation. Thirty-one tumors (60%) were considered "typical oat cell" by electron microscopy and 20 (40%) showed features of epithelial differentiation. Fifteen (75%) tumors with epithelial features were considered operable and nine (45%) were resected with curative intent. In contrast, 26 (84%) tumors considered typical oat cell by electron microscopy presented with extensive metastatic disease. The cancer-free 5-year actuarial survival rate of patients whose tumors showed features of epithelial differentiation was 25%. The actuarial survival rate of nine patients who underwent resection of tumors with epithelial features was 38% at 5 years. Only one patient whose tumor was considered typical of oat cell carcinoma by electron microscopy survived 5 years. Our current recommendation is to remove all clinically resectable pulmonary neoplasms with the expectation that these localized small cell tumors are likely to show epithelial features by electron microscopic analysis. PMID- 2993757 TI - The immunopotentiating effect of epemedii flavone on T-cell function in yang deficient animal model and its relationship with cyclic nucleotides. PMID- 2993758 TI - Study of the action of Ganoderma lucidum on scavenging hydroxyl radicals from plasma. PMID- 2993760 TI - Stereotaxic operation in the neonatal rat; a novel and simple procedure. AB - Since no simple method for standardized positioning of very young rats in a stereotaxic device has been described in the literature, a new method for stereotaxic placement in the neonatal rat was developed, which only requires a simple body mould of the rat pup, and a minor adaptation of a rabbit stereotaxic apparatus (Kopf). The costs of the materials needed for construction of the neonatal rat body mould were approximately $1.-. Construction of the body mould took about one hour. Reliability and accuracy of the new method are demonstrated in a series of experiments in which the A10 group of dopaminergic cell bodies in the ventral tegmental area of 1-day-old rats were lesioned bilaterally: 15 out of 20 angular electrode placements resulted in lesions, the centres of which were located within the A10 group of dopaminergic cell bodies in the ventral tegmental area (max. 0.8 mm in diameter). PMID- 2993759 TI - Purification of non-synaptic and synaptic mitochondria and plasma membranes from rat brain by a rapid Percoll gradient procedure. AB - A method is described for the isolation and purification of non-synaptic and synaptic mitochondria and plasma membranes. The procedure involved differential centrifugation and successively a Ficoll gradient followed by a Percoll one. The method is rapid and maintains constant osmotic conditions. The Percoll-purified fractions show a lower density than those prepared by the conventional methods. The fractions were identified and characterized by enzyme markers and electron microscopy. The method will be useful for routine preparations rendering a good degree of homogeneity and purity. PMID- 2993761 TI - Regression of progressive lymphadenopathy in a young child with acute cytomegalovirus (CMV) infection following the administration of transfer factor with specific anti-CMV activity. PMID- 2993762 TI - Oncogenes. AB - Oncogenes were first identified in neoplastic tissues and were thought to be disseminated by retroviruses. They were shown to cause neoplastic transformation of cells in vitro and were therefore regarded as "the genes that cause cancer." They occur in all eukaryotes including humans. More than 30 oncogenes have been identified, and 28 have already been mapped to the human karyotype. When the protein products of the oncogenes were investigated, they were found to be very similar to known substances that are normally involved in the control of cell division--growth factors, plasma-membrane receptors, modulators of the transduction of exogenous signals through the plasma membrane, and nuclear DNA binding substances. These discoveries changed the original concept of oncogenes. They are now regarded as playing vital roles in the normal control of mitosis, and they should properly be called mitogenes rather than "oncogenes." Like all other genes, if a mitogene is mutated, rearranged, translocated, or otherwise deranged, its effects will change; it may then become an oncogene that will promote disorderly cell division and thus contribute to the pathogenesis of cancer. In addition to providing new insights into the normal control of mitosis and its neoplastic transformation, the discovery of the mitogenes has suggested new approaches to the early diagnosis of cancer, to a more rational classification of neoplasms based on pathogenesis rather than morphology, and, perhaps, to more rational forms of therapy. PMID- 2993763 TI - Glycogen accumulation of the aging human brain. AB - We describe light- and electron-microscopically a new type of intracytoplasmatic inclusions within cell processes of the cerebral cortex and the underlying white matter. These structures measure 5-50 micron in diameter and consist almost exclusively of densely packed alpha- or beta-glycogen granules, which never occur together in any single structure. Within their periphery, electron-dense amorphous spots and cell organelles, especially mitochondria, were seen. No membrane-bound glycogen was observed. We propose to call them granular glycogen bodies. They occur in 4 of 7 examined postmortem specimens of the cerebral cortex of people older than 60 years of age. They were not found in 4 younger controls aged 26-48. Their appearance may reflect a distinct turnover disorder of carbohydrate metabolism, which becomes manifest under diverse pathologic conditions and in the normal aging process. PMID- 2993764 TI - Intestinal morphology, marker enzymes and lipid content of brush border membranes from rabbit jejunum and ileum: effect of aging. AB - Aging is associated with changes in the intestinal uptake of nutrients. This study was undertaken to determine whether the morphology, enzyme markers and the lipid content of the intestinal brush border membrane (BBM) was influenced by aging. There was an increase in the height of the jejunal villi and number of cells/villus, resulting in an age-related increase in the jejunal villus and mucosal surface area in young as compared with weanling rabbits. In mature 1-year old animals, there was a decline in villus height, number of cells/villus, and mucosal surface area, so that the jejunal characteristics of the mature animals resembled those of the weanling rabbits. In the ileum, aging was associated with an increase (weanling vs. young), then a decrease (young vs. mature) in the height of the villi, and the number of cells/villus. Aging had no effect on the size of the villus cells. At all ages there was a direct positive relationship between the height of the villi and the mucosal surface area, and between villus surface area and sucrase activity. An established technique was used to purify rabbit BBM and to measure the BBM content of enzyme markers and lipids in weanling, young and mature animals. Both the BBM sucrase (S) and alkaline phosphatase (AP) increased in young as compared with weanling rabbits, and the ratio of AP/S remained unchanged. The S remained high in mature rabbits, but AP declined, so that AP/S fell. There was a positive linear correlation between S and villus surface area. In weanling rabbits, the total BBM phospholipid content and the ratio of total phospholipid/total cholesterol were lower in the ileum than in the jejunum. In the jejunal BBM of young animals, there was more total free fatty acids and cholesterol ester than in the weanling jejunum. The jejunal BBM total phospholipids and total cholesterol were higher in the mature than in the weanling animal jejunum when expressed as nmoles/mg protein, but the ratio of total phospholipid/total cholesterol was unaffected by aging. The greatest percentage of jejunal BBM phospholipid was comprised of lecithin and phosphatidyl ethanolamine. The increased BBM total phospholipid content in mature animals was associated with a higher amount and lower proportion of lecithin, but a higher proportion of sphingomyelin and phosphatidyl serine.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993765 TI - Age-dependent alterations in rat ventricular myocardium: a quantitative analysis. AB - A quantitative electron microscopic examination of the ventricular myocardium of the male Fischer 344 rat was undertaken to determine the extent of age-dependent changes in ultrastructure. Rats of 6, 16 and 30 months of age were examined using a non-biased stereological test system. Volume fractions of mitochondria, myofibrillar mass and lipid remained unchanged during aging. Dense body volume fraction increased four-fold over the period studied, while the sarcoplasmic reticulum volume increased only from 6 months to 16 months and did not change thereafter. Biochemical analysis of the specific activities of the two lysosomal enzymes, acid phosphatase and beta-glucuronidase demonstrated a differential response to aging, with acid phosphatase remaining constant and beta glucuronidase increasing slightly. Neither enzyme activity correlated with the pronounced change in dense body volume fraction. This study provides clear evidence that the previously observed changes in cardiac performance associated with aging are not readily explicable on the basis of a degradation of cardiac ultrastructure. PMID- 2993766 TI - [Muscular pathology in patients with chronic alcohol consumption]. PMID- 2993767 TI - [Vitamin D: (I) Molecular biology]. PMID- 2993768 TI - [Occurrence, significance and characteristics of human Enteroviruses in Poland 1973-1982]. PMID- 2993769 TI - [Stimulus level dependence of BERA potential amplitudes]. AB - The observed stimulus level dependence of early acoustic evoked potential amplitudes suggests that two different mechanism contribute to neural activity: A first mechanism yields neural activity immediately above the hearing threshold, the activity increasing up to a maximum value which is attained at a stimulus intensity of approximately 40 dB HL. At this level, the onset of a second mechanism is observed, which produces increasing neural activity and exhibits saturation behaviour at high stimulus intensities. A mathematical amplitude intensity characteristic is presented which can be calculated on the basis of simple and plausible assumptions. It is in excellent agreement with the experimental data. Furthermore, in the case of damaged hair cells but undamaged nerve fibres, the mathematical approach leads directly to the steep amplitude characteristic as it is often observed in the case of recruitment. PMID- 2993770 TI - Fluorescein angiography of the tympanic membrane. AB - Little is known about the dynamic vasculature of the tympanic membrane, despite the fact that it is within easy view of the otolaryngologist. An understanding of its blood supply dynamics may lead to a better understanding of the tympanic membrane's physiological properties and response to surgery. Also, certain patterns of blood flow may correlate with disease entities and be of diagnostic value. Currently, we have done 30 fluorescein angiograms of the tympanic membrane. The fluorescein angiograms from a normal ear and from ears with several types of ear pathology are described. PMID- 2993771 TI - Nd-YAG laser for general surgery. AB - We report here our clinical experiences with Nd-YAG laser on general surgery, and evaluate the results of this procedure. From December 1979 to December 1981, we applied Nd-YAG laser to various operations as a hemostatic and cutting tool. For hemostasis, we used conventional quartz fiber which was covered with sterile tube, and hemostatic efficacy was examined especially in the subcutaneous bleeding and the bleeding from solid organs. For cutting, we used special devices, ie, Medilas YAG surgical probe (noncontact-type probe), and a laser blade (contact-type probe), and performed four liver resections. It is concluded that the hemostatic efficacy of Nd-YAG laser to various bleeders was proved in general surgical procedures, and furthermore this laser can cut tissue if we utilize these devices. When comparing these two devices, we would prefer the contact-type probe. PMID- 2993773 TI - Renin release from kidney cortical slices in response to isoproterenol and glucagon is decreased in vitamin E-deficient rats. AB - The effects of vitamin E (VE)-deficiency on renin release by various agents were examined using rat kidney cortical slices. Isoproterenol and glucagon in the presence or absence of theophylline increased renin release in the control group, while their stimulatory effects were attenuated by VE-deficiency. These decreased responses of renin release to isoproterenol and glucagon due to VE-deficiency were restored to the control level by dietary supplementation of dl-alpha tocopheryl acetate or N,N'-diphenyl-p-phenylenediamine. The stimulatory effect of dibutyryl cyclic AMP or theophylline on renin release was not affected by VE deficiency. These results suggest that in case of VE-deficiency, the response of renin release to stimuli is decreased via cyclic-AMP production. PMID- 2993772 TI - Increased activity of 5-lipoxygenase in polymorphonuclear leukocytes from asthmatic patients. AB - The formation of 5-lipoxygenase products of arachidonic acid, 5-HETE and 5,12 diHETE, was determined in 100,000 X g supernatant of polymorphonuclear leukocytes from 17 healthy subjects, 17 patients with extrinsic asthma and 15 patients with intrinsic asthma. After the supernatant was incubated with 14C-arachidonic acid in the presence of calcium and indomethacin, the lipoxygenase products of arachidonic acid were separated by thin layer chromatography. The results were expressed as the percentage conversion of 14C-arachidonic acid into the product per 10(7) cells. The formation of 5,12-diHETE, but not of 5-HETE, was significantly increased in the cells from the group of patients with extrinsic asthma (4.38 +/- 0.78%, mean +/- S.E.; p less than 0.01) and intrinsic asthma (6.09 +/- 1.11%; p less than 0.01), when compared to normal subjects (1.74 +/- 0.30%). Both extrinsic and intrinsic asthmatics had significantly enhanced 5 lipoxygenase activity, which was expressed as the sum of percentage conversion of 14C-arachidonic acid into 5-HETE and 5,12-diHETE. The percentage conversion in normal subjects was 4.19 +/- 0.39%, 6.24 +/- 0.84% for 17 patients with extrinsic asthma (p less than 0.05), and 8.59 +/- 1.29% for 15 patients with intrinsic asthma (p less than 0.01). There was no significant difference between these asthmatic groups. These results indicate that 5-lipoxygenase activity is increased in patients with bronchial asthma. PMID- 2993774 TI - Stimulation of ACTH release by naloxone: central or peripheral action? AB - The concentration of ACTH and corticosterone in plasma were measured following peripheral administration of naloxone and naloxone methylbromide (quaternary derivative of opiate antagonist naloxone which is thought not to cross the blood brain barrier) in male rats. Subcutaneous administration of naloxone methylbromide in the dose range of 0.625 - 5.0 mg kg-1 resulted in a small but significant increase in plasma corticosterone levels. Both naloxone and its quaternary derivative injected via permanent intraperitoneal catheters to freely moving rats induced a highly significant increase in plasma ACTH levels measured in blood obtained via permanent tail artery catheters 30 min following injection. These results indicate that loci outside the blood brain barrier are, at least partially, involved in the naloxone-induced stimulation of ACTH release. PMID- 2993775 TI - The adenylate cyclase-cyclic AMP-protein kinase system in different cell populations of the guinea pig gastric mucosa. AB - In isolated guinea pig gastric mucous and enriched parietal cells it was tested whether or not cyclic AMP in response to histamine stimulation might reach concentrations sufficiently high to activate an intracellular cyclic AMP dependent protein kinase and thereby mediate the acid response. Although histamine stimulated parietal cell adenylate cyclase to a greater extent than mucous cell adenylate cyclase, cyclic AMP levels in response to maximal histamine stimulation reached higher levels in mucous than in parietal cells. This had to be attributed to a five times higher phosphodiesterase activity in parietal cell than in mucous cell populations. In the absence of the phosphodiesterase inhibitor isobutylmethylxanthine exposure of the cells to histamine only in mucous cells produced an increase in cyclic AMP-dependent protein kinase activity ratio, but not in parietal cells. Dibutyryl-cyclic AMP induced cyclic AMP accumulation in parietal cell populations was compared to dibutyryl-cyclic AMP induced H+ secretion, as measured by 14C-aminopyrine uptake. A maximal acid response was associated with an intracellular cyclic AMP level of approximately 300 pmol/10(6) cells, which was never reached by maximal histamine stimulation even not in the presence of the phosphodiesterase inhibitor. It is concluded that activation of the parietal cell cyclic AMP-dependent protein kinase is one way for stimulating H+ secretion, but that the acid response elicited by histamine requires another intracellular pathway. PMID- 2993776 TI - Inhibition of rat cardiac and renal Na+,K+-ATPase by high sodium concentrations and vanadate. AB - In the present study, rat renal Na+,K+-ATPase was found to be more sensitive to inhibition by high Na+ concentrations (100-400 mM) than was rat cardiac Na+,K+ ATPase. K+ was more effective in reversing the inhibition by Na+, of cardiac relative to renal Na+,K+-ATPase. Rat renal Na+,K+-ATPase was also more sensitive than cardiac Na+,K+-ATPase to inhibition by vanadate over this range of Na+ concentrations. These results support the hypothesis that vanadate may selectively regulate Na+,K+-ATPase in the kidney, and they may also help explain the natriuretic and diuretic effects of vanadate in rats. Inhibition of renal Na+,K+ATPase by Na+, may also help explain, in part, the natriuretic and diuretic effects of acute saline loading. PMID- 2993778 TI - Stimulation of adenylate cyclase by adenosine and other agonists in mesenteric artery smooth muscle cells in culture. AB - An adenosine-sensitive adenylate cyclase has been characterized in cultured mesenteric artery smooth muscle cells. N-Ethylcarboxamide-adenosine (NECA), N Methylcarboxamide-adenosine (MECA), L-N6-phenylisopropyladenosine (PIA) and 2 chloroadenosine (2-cl-Ado) all stimulated adenylate cyclase in a concentration dependent manner. NECA was the most potent analog (EC50, 1 microM), whereas PIA (EC50, 15 microM), 2-Cl-Ado (EC50, 15 microM) and MECA (EC50, 24 microM), were less potent and had efficacies relative to NECA of 0.61, 0.61 and 0.65, respectively. Adenosine showed a biphasic effect: stimulation at lower concentrations and inhibition at higher concentrations, whereas 2' deoxyadenosine only inhibited adenylate cyclase activity. The stimulatory effect of NECA on adenylate cyclase was dependent on metal ion concentration and was blocked by 3 isobutyl-l-methylxanthine (IBMX) and 8-phenyltheophylline (8-PT). Adenylate cyclase from these cultured cells was also stimulated by other agonists such as epinephrine, norepinephrine, prostaglandins, dopamine, NaF and forskolin. The stimulation of adenylate cyclase by isoproterenol, epinephrine and norepinephrine was blocked by propranolol but not by phentolamine. On the other hand, phentolamine, propranolol and flupentixol all inhibited dopamine-stimulated adenylate cyclase activity. In addition, the stimulation by an optimal concentration of PIA was additive or almost additive with maximal stimulation caused by catecholamines and prostaglandins. These data indicate the presence of adenosine (Stimulatory "Ra"), catecholamine and prostaglandin receptors in mesenteric artery smooth muscle cells and suggest that these agents may exert their physiological actions through their interaction with their respective receptors coupled to adenylate cyclase. PMID- 2993777 TI - Dopamine-beta-hydroxylase inhibition acutely stimulates rats hypothalamic noradrenaline and dopamine neuronal activity as assessed from metabolic ratios and circulating glucose and ACTH responses. AB - Because central noradrenaline neuronal activity is tonically inhibited by noradrenaline (NA) itself via an action at prejunctional alpha 2-adrenoceptors, it was hypothesised that the blockade of central NA synthesis following acute dopamine-beta -hydroxylase (DBH) inhibition might primarily deplete prejunctional NA levels and result in an increase in central NA neuronal activity through reduced NA autoinhibition. This hypothesis was tested in the rat following the acute administration of the DBH inhibitors diethyldithiocarbamate (DDC) and cysteamine (CSH). Computerised gas chromatography/mass spectrometry was used to precisely measure the hypothalamic levels of NA and dopamine (DA) together with those of their primary neuronal metabolites dihydroxyphenylethyleneglycol (DHPG) and dihydroxyphenylacetic acid (DOPAC), respectively. Both DDC (at 4 h) and CSH (at 30 min.) caused approximately a 50% reduction of hypothalamic NA concentrations. However this was associated with marked and highly significant increases in hypothalamic DHPG levels (by 50-100%) and in the hypothalamic ratio DHPG/NA. Also, when measured after CSH, the hypothalamic levels of the DHPG metabolite 3-methoxy-4-hydroxyphenylethyleneglycol were highly significantly increased. Consistent with increased DA neuronal activity, both DBH inhibitors raised DA and DOPAC levels and also the ratio DOPAC/DA in the hypothalami of treated rats and markedly suppressed serum prolactin levels (all p less than 0.01). The rise in hypothalamic concentrations of DHPG indicates that an increase in hypothalamic NA neuronal activity occurs following DBH inhibition. Significant elevations of blood glucose, corticosterone and ACTH were also observed after DBH inhibition. As we have previously demonstrated that increased central NA activity is associated with elevations of blood glucose, corticosterone and ACTH, these data provide further evidence for a functional increase in central NA activity caused by acute DBH inhibition. It is proposed that the increase in hypothalamic NA activity after DBH inhibition results from a primary depletion of the prejunctional alpha 2-active autoregulatory pool of NA. PMID- 2993779 TI - Polyphosphoinositides and the shape of mammalian erythrocytes. AB - The relationship between polyphosphoinositide and phosphatidic acid (PA) metabolism and Mg-ATP dependent shape and viscosity changes in erythrocyte ghosts from four mammalian species was examined. Ghosts prepared from rabbit, dog, human and guinea pig erythrocytes were transformed from echinocytes to discocytes within 15 min in the presence of 1 mM Mg-ATP at 25 C. In all species these Mg-ATP shape transformations were associated with a 30-45% decrease in the specific viscosity of the ghost suspensions. Mg-ATP induced a second transformation of discocytic ghosts to cup shape forms without a further decrease in viscosity. A considerable species variation in the rates of Mg-ATP dependent viscosity and shape changes and incorporation of 32P into phosphatidylinositol-4' phosphate (PIP), phosphatidylinositol-4'5'bisphosphate (PIP2) and especially PA from Mg [gamma 32P]-ATP in ghosts was found. However, the rates of Mg-ATP dependent synthesis of PIP and PIP2 and shape and viscosity changes in each species were of the same magnitude. Ca2+ or neomycin strongly inhibited PIP labeling and Mg-ATP shape and viscosity changes in ghosts of the different species. Ca2+ or neomycin usually increased or had little effect on 32P incorporation into PA and PIP2. The possibility that Mg-ATP-induced changes in erythrocyte membrane shape and deformability are dependent on increases in membrane PIP and PIP2 is discussed. PMID- 2993780 TI - Phosphorylation of chloroform soluble compounds in plasma membranes of human epidermoid carcinoma A431 cells. AB - This study investigated a possible role for the epidermal growth factor (EGF) receptor protein tyrosine kinase in phosphoinositide metabolism with plasma membrane vesicles from human epidermoid carcinoma (A431) cells. We found a novel chloroform-soluble product radiolabeled with [gamma-32P]ATP that did not migrate from the origin in the thin layer system designed to separate the phosphoinositides, appeared as a single band of Mr = 3500 on polyacrylamide gels in the presence of dodecyl sulfate, had an ultraviolet absorbance spectrum with a maximum at 275 nm and stained with Coomassie dye. Based on these properties this phosphorylation product is referred to as a proteolipid. The 32P label was not detected in phosphotyrosine [Tyr(P)], phosphoserine [Ser(P)] or phosphothreonine [Thr(P)] and was lost during acid or base hydrolysis. Phosphorylation of proteolipid was increased significantly by EGF, whereas phosphorylation of phosphatidic acid was decreased and labeling of phosphoinositides was unaffected. Thus, it appears that in A431 membranes the EGF receptor/kinase does not utilize phosphatidylinositol as a substrate, but does phosphorylate a membrane proteolipid. PMID- 2993782 TI - [Polychemoradiation treatment of non-small cell lung cancer]. PMID- 2993783 TI - Heterogeneity of the erythrocyte Na-K pump status in human obesity. AB - The number of Na-K pump units, the Na-K-ATPase activity, the K transport turnover rate per pump unit and the intracellular Na and K concentrations were measured in the erythrocytes of 56 obese patients and 20 normal subjects. No differences were found between the two groups. In obese patients, we failed to observe any influence of dietary habits, age of onset, or family history of obesity on the Na pump status. On the other hand, we found that the number of pump units was not a close reflection of the membrane cation transport and in some patients with an abnormally high number of pump units, an inappropriately low Na-K-ATPase activity was observed. We also identified two small groups of obese patients with, respectively, abnormally high or low K transport turnover rate per pump unit. Our study seems to support the hypothesis that abnormalities in the erythrocyte Na-K pump system are not usual in the obese population but are probably present only in a limited number of selected patients. PMID- 2993781 TI - [Radionuclide research on reparative bone formation during the treatment of spiral bone fractures of the leg by G. A. Ilizarov's method]. AB - A study was made of reparative osteogenesis and the content of mineral bone components in 60 patients with spiral fractures of the leg bones during treatment by transosseous osteosynthesis. The time of maximum labeled pyrophosphate accumulation was determined by a degree of the initial dislocation of fragments: in the dislocation to the cortical layer it was noted on the 42nd day, by 1/3 on the 45th day, by 2/3 on the 50th day, to the cross section on the 60th day. The demineralization of bone fragments in the dislocation to the cortical layer was 34%, by 1/2 37%, by 2/3 41% and to the cross section by 44%. The labeled sulphuric colloid accumulation by bone marrow cells was maximal in minimal dislocations and decreased with growing dislocation. PMID- 2993784 TI - Purification from rabbit and rat liver of cytochromes P-450 involved in bile acid biosynthesis. PMID- 2993785 TI - Dolichol kinase, phosphatase, and esterase activity in calf brain. PMID- 2993786 TI - Dolichyl pyrophosphate phosphatase in brain. PMID- 2993787 TI - AT-rich linkers in long range organisation of Drosophila DNA. AB - AT-rich segments were mapped in 6 different domains of the Drosophila melanogaster genome by partial denaturation of cloned genomic DNA segments and observation in the electron microscope (EM). As found in a previous investigation (Moreau et al., 1982), three types of AT-rich segments could be distinguished: 1) AT-rich linkers with a very high AT content, 600 +/- 200 bp long; 2) clusters of such AT-linkers extending over up to 10 Kbp and 3) AT-rich stretches which are shorter and of lower AT content compared to AT-linkers. Six genes previously localized in these domains were found to lie in relatively GC-rich segments framed by AT-rich elements of the 3 types: the Larval Serum Protein LSP-1 alpha gene is framed by 2 AT-linkers, the LSP-1 beta and LSP-2 genes by two AT-rich stretches, an the LSP-1 gamma gene by a cluster and a stretch. The 55 Kbp genomic segment encompassing the P6 gene contains and AT-cluster of about 15 Kbp and several AT-linkers and AT-rich stretches. The 85 Kbp domain containing the P1 gene includes 3 AT-rich clusters of about 10 Kbp each framing GC-rich domains punctuated by AT-linkers and stretches. This study shows that AT-mapping allows a rapid diagnosis of large genomic DNA domains in relation to the AT-rich segments which, possibly, are of significance with regard to genome organisation and function. PMID- 2993788 TI - Versatile Escherichia coli-Bacillus shuttle vectors derived from runaway replication plasmids related to CloDF13. AB - Versatile cloning vectors were constructed employing a runaway replication mutant of the bacteriocinogenic plasmid CloDF13. These vectors can, under conditions where protein synthesis is not inhibited, be amplified in Escherichia coli to high levels by elevating the temperature and are therefore useful for the production of large quantities of DNA and protein. Since the constructed shuttle vectors, which harbour at least six unique restriction endonucleases sites, replicate in E. coli, Enterobacter cloacae, Staphylococcus aureus and a variety of Bacilli, they are applicable for the genetic engineering of both gram-negative and gram-positive bacteria. PMID- 2993789 TI - Cointegration and resolution mediated by IS101 present in plasmid pSC101. AB - A certain class of cointegrate plasmids was found to occur between a pSC101 derivative and a second plasmid pBV320 in E. coli F- cells. Cleavage analysis and DNA sequencing showed that the cointegrate plasmid contained direct repeats of an insertion sequence IS101 at the recombination junctions, indicating that formation of cointegrates was mediated by IS101, which is a natural constituent of pSC101. These cointegrates were formed only in cells which contained the transposon gamma-delta, suggesting that the gamma-delta sequence, which provides transposase, is responsible for cointegration. Whenever the cointegrate plasmids were present in cells containing gamma-delta or its related transposon Tn3, the cointegrates were dissolved to give pBV320::IS101 due to recombination at duplicated IS101 sequences in the cointegrates, suggesting that both gamma-delta and Tn3, which provide a resolvase, are responsible for the resolution of the cointegrates. Comparison between the nucleotide sequence of IS101 and those of gamma-delta and Tn3 shows a high degree of homology in the regions that have been shown to be the binding sites of resolvases, as well as in the terminal inverted repeats. However, there is no homology between IS101 and the other element, gamma delta or Tn3, in the internal resolution site, at which the resolution event may occur. PMID- 2993790 TI - Expression of an Escherichia coli phr gene in the yeast Saccharomyces cerevisiae. AB - A 2 kb DNA fragment, containing the photoreactivation gene phr1 from Escherichia coli, was inserted at the BamH1 site in the tet gene of the yeast--E. coli shuttle vector pJDB207. Photoreactivation--deficient Saccharomyces cerevisiae cells transformed with this plasmid showed photoreactivation of killing after UV irradiation of the cells, while extracts of transformed cells exhibited photoreactivating activity in vitro. Far more photoreactivating enzyme molecules were found when the gene was inserted in the plasmid in the opposite orientation to the tet gene as compared with a plasmid carrying the inserted gene in the same orientation. Photoreactivating enzyme encoded by the E. coli phr1 gene and produced in transformed yeast cells has characteristics of the E. coli photoreactivating enzyme (flavoprotein) as judged from the influence of ionic strength on photoreactivating activity. PMID- 2993791 TI - Effects of polymyxin B sulfate and polymyxin B nonapeptide on growth and permeability of the yeast Saccharomyces cerevisiae. AB - Polymyxin B, a toxic, membrane-affecting antibiotic, can be rendered harmless to yeast cells by enzymatic removal of its fatty acyl moiety. The remaining cyclic peptide portion, polymyxin B nonapeptide, has no significant effect on growth and viability but it drastically reduces mating efficiency. In addition, the cyclic peptide enhances sensitivity of cells to several drugs, presumably by increasing membrane permeability. Mutants resistant to polymyxin B are simultaneously less responsive to the combination of the nonapeptide and the drugs. This indicates that the peptide portion of polymyxin B is the moiety responsible for the permeability changes. The resistance is inherited as a simple recessive trait. The mutation has been mapped to chromosome XV of Saccharomyces cerevisiae. PMID- 2993792 TI - Refined genetic analysis of the region II che mutants in Salmonella typhimurium. AB - From a detailed complementation analysis of the region II che mutants of Salmonella typhimurium, we have located five che genes, cheA, cheW, cheR, cheB, and cheY. We have shown that corrections are required in the previous assignment of the mutations in four strains: both SL2514 and SL2515 which have been reported to be cheY mutants are cheR mutants, SL2539 is not a cheA but a cheW mutant, and ST171 which has been reported to be a cheZ mutant is a double mutant with defects in both cheA and cheB. Since ST171 is the only "cheZ mutant" so far isolated, the idea that the cheZ gene might play an essential role in chemotaxis in S. typhimurium as in Escherichia coli has lost its experimental basis. Furthermore, a number of deletion mutants in region II resulting from the excision of Tn10 have been isolated and analysed. From these experiments, we propose that the gene order in region II is flaK-flaE-motA-motB-cheA-cheW-cheR-cheB-++ +cheY-flaM-flaC, which is identical with that in E. coli. PMID- 2993793 TI - Functional complementation between the two homologous genes, ops and tps, during differentiation of Myxococcus xanthus. AB - Protein S is a development-specific protein of Myxococcus xanthus encoded by the tps gene. It has been shown that there are two extensively homologous genes (ops and tps) tandemly repeated in the same direction with a 1.4 kb spacer fragment between them (Inouye et al. 1983). Seven deletion mutants were constructed by removing the ops gene, the tps gene, segments of the spacer sequence or combinations of these regions. The deleted regions were replaced with DNA fragments carrying the Tn5 gene for kanamycin resistance. The effects of deleting different regions on morphological changes and on patterns of protein synthesis during fruiting body formation were examined. The process of fruiting body formation was severely delayed when both the ops and the tps genes were deleted. However, this delay could be suppressed by either the ops gene or the tps gene, individually, although in the latter case, a slight delay was still observed. These results indicate that the ops gene is expressed during fruiting body formation and plays a role in the normal program of M. xanthus differentiation. Furthermore, the role of the ops gene can be complemented by the tps gene. The deletion of the ops and/or tps genes had no effect on glycerol-spore formation. PMID- 2993794 TI - Cloning of the ARO cluster gene of Neurospora crassa and its expression in Escherichia coli. AB - We have constructed a phage, lambda Ncl, which comprises a 4.0 kb HindIII insert of Neurospora DNA into the immunity region of the vector lambda 598. lambda Ncl complements the aroD6 mutation of E. coli, permitting the formation of galaxy plaques on medium lacking aromatic supplements, and transforms an aro-9 qa-2 Neurospora mutant to prototrophy at a low frequency. Low levels of 5 dehydroquinate hydrolyase (E.C.4.2.1.10.), with properties unlike those of the catabolic isoenzyme that is coded by qa-2, are present in E. coli aroD6 cell lysates following infection with lambda Ncl. lambda Ncl does not hybridize with qa-2 DNA and it is concluded that it contains at least the aro-9 region of the pentafunctional aro cluster gene. PMID- 2993795 TI - Comparative sequence and functional analysis of pT181 and pC221, cognate plasmid replicons from Staphylococcus aureus. AB - The nucleotide sequence of pC221, a 4.6 kb Staphylococcus aureus plasmid is presented. The replication region of the plasmid is identified and compared with the corresponding region of pT181, a compatible but related plasmid. Both plasmids encode trans-active replicon-specific initiator proteins, RepC for pT181 and RepD for pC221. Plasmid replication rate is controlled by regulation of the rate of synthesis of the initiator protein by means of inhibitory 5' countertranscripts. Key elements of the control system are closely conserved between the two plasmids whereas less critical elements show extensive divergence. Overall architecture is also conserved, suggesting functional parallelism. The replication origin for both plasmids is contained within the N terminal region of the initiator protein coding sequence; the two coding sequences are highly homologous but have two important areas of divergence, one within the origin region, the other near the C-terminus. In vivo recombinants between the two plasmids isolated previously (Iordanescu 1979) have crossover points within the initiator gene, between the two divergent regions. The recombinant plasmids have hybrid initiator proteins and are defective for replication, requiring the simultaneous presence of the parental plasmid from which their origin is derived. They are able to complement replication-defective mutants of the other parental plasmid, suggesting that the recognition specificity of the hybrid initiator protein resides in its C-terminal end and that the specific recognition site for the protein corresponds to the divergent region within the origin. PMID- 2993796 TI - Recombination in the lambda repressor gene: evidence that very short patch (VSP) mismatch correction restores a specific sequence. AB - The mutation am6 in the cI gene of bacteriophage lambda is identified as a C----T transition in a 5'CCATGG sequence. In four-factor crosses of am6 with nearby mutations in cI, the frequencies of cI+ recombinants are much higher than expected from the physical distances. A very short patch (VSP) mismatch repair system is presumed to recognize am6/am+ mispairs in the heteroduplexes that accompany recombination between the outside markers. Mutation am6 is corrected to am+; correction of am+ to am6 was not detected. Clear-plaque mutation 1-1 in cI is a T----C transition in a 5'CTTGG sequence, resulting in the sequence 5'CCATGG. When 1-1 was crossed with nearby mutations in gene cI, there were no excess cI+ recombinants, which would result from repair of CCTGG (1-1) to CTTGG (cI+). However, in crosses of cI+ phages with mutation 1-1, there was an excess of cI- recombinants, indicating that cI+ was repaired to 1-1. Preferential repair does not require adenine or cytosine methylation: when repairing a mismatch, the VSP repair system apparently identifies specific mispaired bases by sequence alone. PMID- 2993797 TI - Cloning of sporulation gene spoIVC in Bacillus subtilis. AB - Sporulation gene spoIVC of Bacillus subtilis was cloned by the prophage transformation method in temperate phage phi 105. The specialized transducing phage, phi 105 spoIVC-1, restored the sporulation of the asporogenous mutant of B. subtilis strain 1S47 (spoIVC133). Transformation experiments showed that the spoIVC gene resides on a 7.3 kb HindIII restriction fragment. Subsequent analysis of the 7.3 kb HindIII fragment with restriction endonuclease EcoRI showed that the spoIVC gene resides on a 3.6 kb EcoRI fragment within the 7.3 kb fragment. The 3.6 kb fragment was recloned into the unique EcoRI site of plasmid pUB110 and deletion derivatives having a deletion within the 3.6 kb insert were constructed. The plasmid carrying the entire spoIVC gene restored the sporulation of strain HU1214 (spoIVC133, recE4) at a frequency of 10(7) spores/ml, and reduced the sporulation of strain HU1018 (spo+, recE4) to 10(7) spores/ml. PMID- 2993798 TI - An electron microscopic heteroduplex study of the sequence relations between the bacteriophages LP52 and theta. AB - The genomes of the phylogenetically related but morphologically distinct bacteriophages LP52 and theta (theta) were compared by electron microscopic heteroduplex analysis. The heteroduplex maps were aligned with known restriction maps. In the heteroduplices of LP52 DNA (63.8 kb) with the DNA of the lytic phage theta c (65.9 kb) the tracts of homologous DNA cover about 50% of the genome length and are interspaced by four large and ten smaller non-base-paired regions. The largest block of non-homologous DNA (18.9 kb), represents the right-hand end and there is an unmatched piece of DNA at the left-hand end as well. Most of the heterology is due to substitution resulting in the conservation of the total length of DNA; the three insertions/deletions amount to less than 3.2% of the genome length. Heteroduplices between the DNAs of phage LP52 and the temperate phage theta 1 (65.0 kb) resembled those of LP52:theta c except for the absence of minor loops. Heteroduplex theta c:theta 1 displayed about 9% heterology in seven separate loops which coincided with sections of diversity on the restriction maps; 4.8% of theta 1 DNA did not hybridize with either theta c or LP52 DNA. PMID- 2993799 TI - Characterization of the 3' end of the leucine operon of Salmonella typhimurium. AB - The nucleotide sequence of the leuD gene of Salmonella typhimurium and of the downstream flanking region are presented. S1 mapping experiments identified 3' endpoints of leu mRNA 140 and 285 nucleotides downstream of the UAA stop codon of leuD mRNA. Experiments employing pulse-labeled RNA suggest that these endpoints result from transcription termination rather than RNA processing. Our results indicate that the organization of the 3' non-translated region of the leu operon from S. typhimurium resembles that of the trp operon of Escherichia coli. Further, our results suggest that the leu operon of S. typhimurium does not contain structural genes other than those identified by genetic experiments, i.e. leu, A,B,C and D. PMID- 2993801 TI - Regulation of the expression of iso 2-cytochrome c gene in S. cerevisiae: cloning of the positive regulatory gene CYP1 and identification of the region of its target sequence on the structural gene CYP3. AB - CYP1 is a trans acting regulatory locus modulating both iso 1- and iso 2 cytochrome c synthesis. Genetical analysis of various mutated alleles has allowed us to identify the gene product as a positive regulatory element. The region of the target sequence of the CYP1 product on the iso 2-cytochrome c structural gene was located by molecular and genetic analysis of two cis acting mutations located at the CYP3 locus: CYP3-36 and CYP3-4, which have been shown to arise from the integration of TY1 elements near the promoter site. Determination of the amount of iso 2-cytochrome c synthesized by strains bearing various genetic constructions, in which the cis acting mutations were associated with different alleles of the CYP1 trans acting locus, showed that TY1 inserted into CYP3-36 extinguishes the activation function due to a mutated overproducer allele CYP1 18, while CYP3-4 amplifies this function. This result identifies at least a part of the target sequence of the CYP1 product within the region separating the two TY1 insertions. To clone the CYP1 gene, we took advantage of the iso 2-cytochrome c overproducer phenotype of the mutated allele CYP1-18, which confers a Lactate+ phenotype on an iso 1-cytochrome c-deficient strain. Such a phenotype allowed the isolation of a recombinant plasmid YEpJFM1 carrying the mutated allele, able to complement on lactate medium a lactate- recipient strain. The identity of the YEpJFM1 sequence with the chromosomal gene was confirmed by homologous recombination at the CYP1 locus. PMID- 2993800 TI - Rec-dependent and Rec-independent recombination of plasmid-borne duplications in Escherichia coli K12. AB - Plasmidic recombination in E. coli K12 has been previously demonstrated to be dependent on the host rec genotype. The construction of plasmids that carry a duplication within an antibiotic-resistance gene is described. Recombination between the direct repeats recreates an active antibiotic-resistance gene, allowing quantitative analysis of recombination frequencies in a closely related set of E. coli K12 strains carrying various rec mutations. Using this system, intraplasmidic recombination of a duplication within the pBR322 tetracycline resistance gene is shown to be rec-dependent while recombination of a similar duplication within the kanamycin-resistance gene of Tn903 is shown to be independent of recA, recB, recC, recE, recF and sbcB. PMID- 2993802 TI - An active variant of the prokaryotic transposable element IS903 carries an amber stop codon in the middle of an open reading frame. AB - The prokaryotic mobile genetic element IS903.B is an active variant of IS903. It differs from IS903 and IS102 by 34 and 61 nucleotide substitutions, respectively. The large open reading frame (ORFI) which probably encodes the transposase is conserved in all three IS elements, whereas the smaller open reading frame (ORFII), which codes on the opposite DNA strand and entirely overlaps ORFI, contains an amber stop codon past the middle of ORFII in IS903.B. Experiments using Escherichia coli K12 strains permissive or non-permissive for amber mutations revealed no difference in the cointegration frequency mediated by IS903.B. Therefore, a possible peptide encoded by ORFII on the IS903-related element is unlikely to be necessary for transposition. PMID- 2993803 TI - A temperature-sensitive mutation constructed by "linker insertion" mutagenesis. AB - The in vitro mutagenesis of cloned DNAs allows the formation of virtually any specific mutation, but no method has been found which might routinely lead to the important phenotype of temperature sensitivity. We have studied three linker insertion mutations in the envelope gene of Moloney murine leukemia virus (M MuLV), and found that one was exquisitely temperature-sensitive for plaque formation. We suggest that the construction of short insertion mutations may be a fruitful approach for the generation of temperature-sensitive phenotypes in cloned genes. PMID- 2993805 TI - Function of ribonuclease H in initiation of DNA replication in Escherichia coli K 12. AB - Escherichia coli rnh mutants lacking ribonuclease H (RNase H) activity can tolerate deletion of the origin of DNA Replication (delta oriC) and transposon insertional inactivation of an initiator gene (dnaA::Tn10). Introduction of the recA200 allele encoding a thermolabile RecA protein into rnh- dnaA::Tn10 and rnh- delta oriC mutants strains rendered DNA synthesis and colony formation of these mutants temperature sensitive. The temperature sensitivity and the broth sensitivity (Srm-) of the rnh- dnaA::Tn10 recA200 strain was suppressed by the presence of plasmids (pBR322 derivatives) carrying dnaA+ only when the intact oriC site was present on the chromosome. Lack of RNase H activity neither promoted replication of minichromosomes (pOC24 and p lambda asn20) in the absence of required DnaA+ protein nor inhibited dnaA+-dependent minichromosome replication. These results led to the conclusion that RNase H is not directly involved in the events leading to initiation of DNA replication at oriC. Rather, it functions as a specificity factor by eliminating certain forms of RNA-DNA hybrids which could otherwise be used to prime DNA replication at sites other than oriC. PMID- 2993804 TI - Organization of the MAL loci of Saccharomyces. Physical identification and functional characterization of three genes at the MAL6 locus. AB - We have physically and functionally identified three genes at the MAL6 locus of Saccharomyces carlsbergensis. Using multicopy yeast plasmid vectors, we have subcloned various segments of the entire MAL6 locus. The functional characterization of the MAL6 subcloned regions was determined by (1) analyzing biochemically the levels of MAL-encoded proteins (maltase [alpha-D-glucosidase, E.C. 3.2.1.20] and maltose transport protein) in cells transformed with various MAL6 subclones, and (2) testing the ability of the subclones to complement the maltose fermentation defects of well characterized Mal- mutants in the highly homologous MAL1 locus. The physical homology between MAL6 and MAL1 is in part demonstrated by the gene disruption of MAL1 using subcloned MAL6 DNA sequences. The results demonstrate that the MAL6 locus is a complex of at least three genes: MAL6R, MAL6T and MAL6S. These genes specify, respectively, a regulatory function, a maltose transport activity (presumably the maltose permease) and the structural gene for maltase. The functional organization of the MAL6 locus is thus identical to that which we had previously determined by mutational analysis for the MAL1 locus. PMID- 2993806 TI - Nucleotide sequence of the iron regulatory gene fur. AB - The fur gene of Escherichia coli is involved in all iron-regulated transcriptions hitherto studied. The nucleotide sequence of an 868 basepair fragment containing the fur gene was determined. There was only a single longer reading frame. The amino acid sequence derived from the nucleotide sequence comprised 148 amino acids that together made a polypeptide of 16,795 daltons. The amino acid sequence was confirmed by determination of the amino acid composition, the carboxy terminal lysine residue and the internal Lys-Lys and Lys-Arg sequences of the isolated Fur protein. The nucleotide sequence contains typical initiation and termination sites for transcription and translation. PMID- 2993807 TI - DNA homology and adsorption specificity of Pseudomonas aeruginosa virulent bacteriophages. AB - The DNA homology and adsorption specificity of newly isolated virulent bacteriophages of P. aeruginosa have been studied. On the basis of this analysis all phages were divided into four groups: phi k, phi m, phi mnP78-like and phi mnF82-like bacteriophages. DNA's of phi k as well as phi m phages were shown to possess different restriction patterns although they have an extensive homology. Unlike other groups, phi k phages were characterized by the presence of T4 DNA ligase--repaired, single-chain breaks. PMID- 2993808 TI - Relationship between the proteins encoded by the exclusion determining locus of the IncI plasmid R144 and the cellular localization of these proteins in Escherichia coli K-12. AB - A region of the IncI plasmid R144, determining and controlling exclusion (exc), codes for two proteins, designated 13K and 19K after their apparent molecular weights (respectively 13,000 and 19,000). Both proteins were simultaneously affected by various mutations that resulted in exclusion deficiency. In this paper the relationship between these proteins as well as their cellular location is reported. We found no indications that the 19K protein is a precursor form of the 13K protein. Analysis of gene products of recombinant plasmids carrying exc as well as of several derivatives, however, provided a strong indication that the proteins result from overlapping genes. Besides, evidence was obtained that the 19K protein is essential for exclusion. Localization studies revealed that this protein exists in a membrane-bound form, associated at the periplasmic side of the inner membrane, and in a soluble form residing in the cytoplasm. PMID- 2993809 TI - Purification of the NusB gene product of Escherichia coli K12. AB - The nucleotide sequence of the entire nusB gene of Escherichia coli has recently been determined and the amino acid sequence of its product deduced (Ishii et al. 1984; Swindle et al. 1984). The NusB protein was purified by chromatography on Sephadex G-100, phosphocellulose and hydroxylapatite. Purification of the protein was monitored using 14C-labelled NusB protein, which was synthesized in a maxicell containing an nusB plasmid as a marker. The final product, which was at least 95% pure as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, had a molecular weight of about 16,000 and an isoelectric point of about 7.3. Analytical data on the amino acid composition of the purified protein agreed with that deduced from the DNA sequence and indicated that this protein was indeed the product of the nusB gene. PMID- 2993811 TI - Tn1721-encoded resolvase: structure of the tnpR gene and its in vitro functions. AB - A 760 base pair nucleotide sequence of transposon Tn1721 containing the resolvase (tnpR) gene has been determined. A 186 triplet open reading frame was assigned to tnpR, and the allocation of -35 and -10 promoter boxes was supported by mapping transcription initiation at 70 base pairs upstream of tnpR. Expression of tnpR under tac promoter control generated sufficient resolvase protein for enzyme purification and for in vitro studies. Purified Tn1721 resolvase requires supercoiling and two directly oriented resolution (res) sites. The enzyme resolves cointegrate substrates containing repeat copies of Tn1721 res, of Tn21 res, of Tn21 res/Tn1721 res, but not of Tn3 res. PMID- 2993812 TI - Negative control of oriC plasmid replication by transcription of the oriC region. AB - We have demonstrated that the replication of the oriC plasmid, carrying the replication origin of the Escherichia coli chromosome, is inhibited by transcriptional readthrough from an oriC flanking region of the plasmid. This was drawn from an examination of the replication of an oriC plasmid, pKZ4, which bears the lacOP segment at the right-hand side of oriC (the asnA side) in such an orientation that transcription from the lac promoter proceeds towards oriC. Replication of pKZ4 was found to be drastically inhibited by inducing transcription from the lac promoter with IPTG, an inducer of the lactose operon. When trp transcription attenuator termination sequences were inserted near the right-hand end of the oriC region of pKZ4, the replication of the plasmid became considerably insensitive to the inhibitory effect of IPTG. This indicates that the inhibition is due to the frequent leftward transcription, which initiates at the lac promoter and proceeds into the oriC region. Since IPTG inhibits the replication of pKZ4, but not that of another coexisting oriC plasmid which is devoid of the lacOP segment, the replication inhibition is judged to act only in cis. Transcription from the promoter of the chloramphenicol resistance gene also caused the inhibition of oriC plasmid replication. PMID- 2993810 TI - The resolvase protein from the transposon Tn21. AB - The tac promoter was inserted into Tn21 upstream of the tnpR gene and the resultant plasmid was used to generate substantial amounts of resolvase. This protein was purified to homogeneity. The protein was characterized by amino acid sequence studies (which showed that an open-reading frame previously identified by DNA sequencing had been correctly assigned to the tnpR gene) and by molecular weight measurements (which demonstrated that the only active for of the protein in solution was dimeric). Pure Tn21 resolvase catalysed site-specific recombinations between directly repeated res sites from Tn21 or Tn1721 but not from Tn3 nor on inverted res sites from Tn21. PMID- 2993813 TI - Nucleotide sequence of a spectinomycin adenyltransferase AAD(9) determinant from Staphylococcus aureus and its relationship to AAD(3") (9). AB - The nucleotide sequence of the spc determinant of the Staphylococcus aureus transposon Tn554 has been determined. This gene encodes a spectinomycin adenyltransferase, AAD(9), that mediates resistance to spectinomycin but not to streptomycin. The sequence predicts a 260 amino acid protein of molecular weight 28,943. A spectinomycin-sensitive mutant (spc-1) contains a G----A transition resulting in substitution of threonine (ACA) for alanine (GCA) at residue 165. The predicted amino acid sequence is 36% homologous to that of a widely distributed, gram-negative streptomycin/spectinomycin adenyltransferase, AAD(3") (9), specified by the aadA determinant (Holingshead and Vapnek 1985). PMID- 2993815 TI - Transposon donor plasmids, based on ColIb-P9, for use in Pseudomonas putida and a variety of other gram negative bacteria. AB - The properties of pLG221, a derivative of the ColIb plasmid carrying the transposon Tn5 are described. This plasmid can be used to introduce Tn5 by conjugation from Escherichia coli into a variety of Gram negative bacteria outside the host range for maintenance of ColIb. Plasmid pLG221, and a similar plasmid pLG223 carrying Tn10 may be of general utility as vectors for transposon mediated mutagenesis in a variety of Gram negative bacteria. PMID- 2993814 TI - Structure of genes and an insertion element in the methane producing archaebacterium Methanobrevibacter smithii. AB - DNA fragments cloned from the methanogenic archaebacterium Methanobrevibacter smithii which complement mutations in the purE and proC genes of E. coli have been sequenced. Sequence analyses, transposon mutagenesis and expression in E. coli minicells indicate that purE and proC complementations result from the synthesis of M. smithii polypeptides with molecular weights of 36,697 and 27,836 respectively. The encoding genes appear to be located in operons. The M. smithii genome contains 69% A/T basepairs (bp) which is reflected in unusual codon usages and intergenic regions containing approximately 85% A/T bp. An insertion element, designated ISM1, was found within the cloned M. smithii DNA located adjacent to the proC complementing region. ISM1 is 1381 bp in length, has 29 bp terminal inverted repeat sequences and contains one major ORF encoded in 87% of the ISM1 sequence. ISM1 is mobile, present in approximately 10 copies per genome and integration duplicates 8 bp at the site of insertion. The duplicated sequences show homology with sequences within the 29 bp terminal repeat sequence of ISM1. Comparison of our data with sequences from halophilic archaebacteria suggests that 5'GAANTTTCA and 5'TTTTAATATAAA may be consensus promoter sequences for archaebacteria. These sequences closely resemble the consensus sequences which precede Drosophila heat-shock genes (Pelham 1982; Davidson et al. 1983). Methanogens appear to employ the eubacterial system of mRNA: 16SrRNA hybridization to ensure initiation of translation; the consensus ribosome binding sequence is 5'AGGTGA. PMID- 2993816 TI - Identification of the gene appA for the acid phosphatase (pH optimum 2.5) of Escherichia coli. AB - A strain of Escherichia coli exhibiting reduced activity of the periplasmic enzyme acid phosphoanhydride phosphohydrolase (pH 2.5 acid phosphatase) was isolated. The mutation designated appA1 was located at 22.5 min on the E. coli genetic map. Acid phosphatase purified from an appA- transductant showed less than ten percent of the specific activity of an isogenic appA+ strain. The mutant enzyme was highly thermolabile and its Km for paranitrophenyl phosphate was increased about 20-fold. The mutant protein cross-reacted with antibody to the wild-type enzyme and had the same molecular weight and concentration in extracts as the wild-type enzyme. These findings strongly suggest that appA is the structural gene of the acid phosphatase. PMID- 2993817 TI - ISL1: a new transposable element in Lactobacillus casei. AB - The genome structures of a temperate Lactobacillus phage, phi FSW, and its virulent mutants, phi FSVs, were examined by restriction, heteroduplex and nucleotide-sequence analyses. The results showed that two out of three phi FSVs had the same 1.3 kbp insertion (designated as ISL1) at different positions in the phi FSW sequence. ISL1 was 1,256 bp long and contained at least two long open reading frames of 279 and 822 bases on one strand. Inverted repeats were found at the termini of the ISL1 which was bracketed by 3 bp direct repeats of the phi FSW sequence. From this evidence, we concluded that ISL1 was a transposable element in Lactobacillus casei. PMID- 2993818 TI - The DNA sequence of the gene for the secreted Bacillus subtilis enzyme levansucrase and its genetic control sites. AB - We present the sequence of a 2 kb fragment of the Bacillus subtilis Marburg genome containing sacB, the structural gene of levansucrase, a secreted enzyme inducible by sucrose. The peptide sequence deduced for the secreted enzyme is very similar to that directly determined by Delfour (1981) for levansucrase of the non-Marburg strain BS5. The peptide sequence is preceded by a 29 amino acid signal peptide. Codon usage in sacB is rather different from that in the sequenced genes of other secreted enzymes in B. subtilis, especially alpha amylase. Genetic evidence has shown that the sacB promotor is rather far from the beginning of sacB (200 bp or more). The 200 bp region preceding sacB shows some of the features of an attenuator. A preliminary discussion of the putative workings and roles of this attenuator-like structure is proposed. sacRc mutations, which allow constitutive expression of levansucrase, have been located within the 450 bp upstream of sacB. It is shown that sacRc and sacR+ alleles control in cis the expression of the adjacent sacB gene. PMID- 2993819 TI - Physical and genetic analysis of IS110, a transposable element of Streptomyces coelicolor A3(2). AB - On at least three independent occasions a 1.6 kb segment of Streptomyces coelicolor DNA was detected in apparently the same location in an attP-deleted derivative of the temperate phage phiC31 that carried a selectable viomycin resistance gene. This sequence (termed IS110) allowed integration of the phage (giving viomycin-resistant transductants) at homologous sequences (detected by Southern hybridisation) at several locations in the S. coelicolor genome. The inserted prophages facilitated genetic mapping of two IS110 copies in the chromosomal linkage map. A third copy did not exhibit simple segregation with chromosomal markers, and there appeared to be a frequent DNA rearrangement close to this copy. Some variation in the number of copies of IS110 and their location has taken place in the pedigree of S. coelicolor derivatives. IS110 did not hybridise to any known S. coelicolor plasmid, nor to any of several other IS-like elements previously described in other Streptomyces plasmids or phages. It hybridised strongly to DNA from only a small minority of other Streptomyces species and was absent from S. lividans, a close relative of S. coelicolor. PMID- 2993820 TI - Molecular analysis of P element behavior in Drosophila simulans transformants. AB - In this report we describe the successful transformation of Drosophila simulans with an autonomous P element from Drosophila melanogaster without the use of a selectable marker. This result demonstrates that there is no species barrier for P element transposition. Utilizing gel blotting and in situ hybridization techniques, we have monitored the behavior of newly-introduced P elements in several D. simulans transformed lines over twelve generations. In most instances, an overall increase in the number of P elements was observed. An examination of the frequency of P-element-bearing individuals in one line revealed the rapid spread of P elements through the population. Analysis of well-characterized sublines confirmed that P elements increase in number by transposition to new genomic sites. The formation of degenerate elements occurred in at least one case. These observations suggest that P elements may behave similarly in D. melanogaster and D. simulans. PMID- 2993821 TI - The recQ gene of Escherichia coli K12: molecular cloning and isolation of insertion mutants. AB - The recQ gene of Escherichia coli K12 was subcloned from plasmid pKO1 (Oeda et al. 1981) by monitoring the capacity of the resulting recombinant plasmids partially to reverse the increased ultraviolet (UV) sensitivity of a recF143 recQ1 double mutant. We were able to trace this complementation activity to a 3.4 kilobase (kb) SalI-PvuII fragment. Furthermore, analysis of the Tn3 insertion mutations that abolished the complementation revealed the exclusive localisation of such insertions in the same 3.4 kb segment. This segment was situated about 4 kb clockwise from corA on the chromosome, a result consistent with the transductional data previously reported. In addition, a comparison of our restriction endonuclease cleavage map with the published data has placed recQ between pldA and pldB. When relocated to the recQ site on the chromosome, the recQ::Tn3 mutations conferred partial resistance to thymineless death (TLD) or, in the case of a recBC sbcB background, recombination deficiency and increased UV sensitivity. This has provided the firm evidence that both the TLD resistance and the deficiency in the RecF recombination pathway result from loss of the functional recQ gene. We also identified the recQ gene product as a 74 kilodalton polypeptide by using the maxicell technique. PMID- 2993822 TI - Dispensability of either penicillin-binding protein-1a or -1b involved in the essential process for cell elongation in Escherichia coli. AB - A strain of Escherichia coli lacking the entire ponB gene and a strain lacking the proximal part of the ponA gene were constructed by substitution with a drug resistance gene. These strains lost either penicillin-binding protein(PBP)-1b or 1a totally and their growth was apparently normal at 30 degrees C and 42 degrees C except that growth of the ponB deletion strain was poor on a nutrient agar plate containing no NaCl at 30 degrees C as well as at 42 degrees C. Transductional experiments to introduce the ponB deletion into the ponA deletion strain, and vice versa, showed that the ponA ponB double deletion was lethal unless the deletion was functionally compensated, e.g., by the presence of a plasmid carrying either gene. Thus, either PBP-1b (ponB) or PBP-1a (ponA), but not both, is dispensable for cell viability, at least under ordinary culture conditions. Transductional experiments also suggested that the gamma component of PBP-1b or the PBP-1b lacking the C-terminal portion encoded in the distal region to the SphI site on the ponB was sufficient for supporting growth of the E. coli cell. PMID- 2993823 TI - Vectors for transposon mutagenesis of non-enteric bacteria. AB - We have constructed a series of transposon delivery vectors derived from pRK2013. Since pRK2013 has a broad host range transfer system and a ColE1 replicon, it can be transferred to, but not replicated in, many non-enteric gram-negative bacteria. Thus pRK2013 provides an effective mechanism for the transient introduction of a transposon. Delivery vectors containing Tn7 (tmp str), Tn10 (tet), Tn10 HH104 (tet), or Tn5-132 (tet) have been constructed. When transposition in Caulobacter crescentus was examined, both Tn7 and Tn5-132 were found to transpose efficiently. In contrast, although the antibiotic resistances of Tn10 and Tn501 (mer) were expressed in C. crescentus, no transposition was observed with either transposon. However, transposition of Tn10 from the Tn10 vectors did occur in Acinetobacter calcoaceticus, and transposition of Tn501 from pMD100 has been demonstrated in Rhizobium japonicum (Bullerjahn and Benzinger 1984). Thus, transposon-host interactions play an important role in the determination of whether a particular transposon can transpose in a given host. Furthermore, the results with C. crescentus indicate that there must be different requirements for host interactions for Tn10 and Tn501 than for Tn5 and Tn7. PMID- 2993824 TI - Molecular cloning and functional analysis of the cysG and nirB genes of Escherichia coli K12, two closely-linked genes required for NADH-dependent nitrite reductase activity. AB - We have cloned two genes, nirB+ and cysG+ which are required for NADH-dependent nitrite reductase to be active, from the 74 min region of the Escherichia coli chromosome. Restriction mapping and complementation analysis establish the gene order crp-nirB-cysG-aroB. Both genes are trans-dominant in merodiploids and, under some conditions, can be expressed independently. The cysG+ gene can be expressed from both high and low copy number plasmids carrying a 3.6 kb PstI EcoRI restriction fragment. Attempts to sub-clone the nirB+ gene into pBR322 on a 14.5 kb EcoRI fragment were unsuccessful, but this fragment was readily sub cloned into and expressed from the low copy number plasmid pLG338 (Stoker et al. 1982). Overproduction of the 88 kDa nitrite reductase apoprotein by strains carrying a functional nirB+ gene suggests that nirB is the structural gene for this enzyme. PMID- 2993825 TI - Effect of cyclosporin A on natural killer cells' response during viral infections. AB - In the this study the modification of lymphocyte subsets (T3, T4, T8) and Natural Killer (NK) cells in organ transplanted patients treated with Cyclosporin A (CyA) in the course of viral infection, have been analyzed. Different subsets have been studied with the monoclonal antibody method and infective processes have been verified by serological data of seroconversion. Our study has shown that CyA at the adopted doses does not alter NK response to viral infection; in fact, in patients with seroconversion, higher NK values and lower OKT4/OKT8 ratio values have been found with respect to patients who did not show any viral infection serologic data. Furthermore an increased incidence of reject crisis has been observed in patients with seroconversion. PMID- 2993826 TI - Antiviral activity of coumermycin: identification of resistant and sensitive retrovirus strains. AB - The effect of Coumermycin (CA1) on the replication of different ecotropic retroviruses was studied. Most strains showed reductions in infectivity of less than ten fold, while two strains demonstrated a marked sensitivity to the drug. The isolation of one resistant strain could suggest a specific CA1 antiviral activity. PMID- 2993827 TI - Herpes simplex virus infection in patients affected by dental caries. AB - Herpes simplex virus (HSV) excretion was studied in patients with dental diseases. Two groups of patients were examined: the first with dental caries and the second without dental decay. Results showed a significantly higher elimination rate of HSV in dental caries patients in comparison with control patients. PMID- 2993828 TI - Gene structures of low-neurovirulent vaccinia virus LC16m0, LC16m8, and their Lister original (LO) strains. AB - Vaccinia viruses LC16m0 and LC16m8 are temperature-sensitive and low neurovirulent variants derived from the Lister (Elstree) (LO) strain. Analyses of genome DNAs by digestion with restriction endonucleases and cross-hybridization of the digested fragments revealed that LC16m0 and LC16m8 possess a new XhoI site in addition to the 14 XhoI sites of LO. This new site is located at about 12 X 10(6) daltons from the right terminal end. There was no significant difference in the genome structures between the LC16 variants and LO except the new XhoI site and their terminal fragments which were not identified in LO owing to their heterogeneity. With HindIII digested fragments, there was no difference among the three viruses. This complete mapping raised the possibility that the putative gene responsible for temperature sensitivity and neurovirulence is located at the region of the XhoI site found in LC16m0 and LC16m8. PMID- 2993830 TI - The cell biology of leukaemia. PMID- 2993829 TI - Abnormal glucose metabolism in murine cytomegalovirus (MCMV)-infected mice. AB - Five strains of male and female mice were infected with salivary gland-passaged murine cytomegalovirus (SG-MCMV) intraperitoneally. Although none of them became hyperglycemic, 13-30% of BALB/c male and female, ICR/Slc male and female and C57BL/6J male mice had abnormal values in glucose tolerance tests (GTT) 14 to 30 day post infection. Serum insulin levels in BALB/c male mice were low and infective virus was detected in the pancreas during the acute phase of infection. Histopathology showed mild edematous changes with inflammatory cell infiltrations in the pancreas. Viral antigen was located in acinar cells and occasionally in beta cells by an immunofluorescence test. PMID- 2993831 TI - HTLV-III antibody in Sydney homosexual men. PMID- 2993832 TI - Anti-HTLV-III antibody incidence. PMID- 2993833 TI - Neutralising antibody against type 1 and type 2 herpes simplex virus in cervical mucus of women with cervical intra-epithelial neoplasia. AB - Patients with cervical intra-epithelial neoplasia had significantly increased neutralising antibody activity to type 2 herpes simplex virus in the cervical mucus. While patients differed from control subjects with respect to their number of sexual partners and socio-economic class, there were significant differences in neutralising antibody activity for case control comparisons within the same number of sexual partners or socio-economic groupings. The results lend support to the putative association between type 2 herpes simplex virus infection and pre invasive and invasive carcinoma of the uterine cervix. PMID- 2993834 TI - Assay of extracellular proteinases using a colorimetric collagen substrate for the differentiation of Serratia in the tribe Klebsielleae. AB - The gelatin test has been utilized for many years as a characteristic to separate the genus Serratia from other members of the tribe Klebsielleae. Gelatin is a large protein matrix that cannot diffuse into bacterial cells. Microbes that attack gelatin do so by producing extracellular proteinases. The measurement of gelatinase has suffered from the lack of a definable endpoint and the inability to quantitate the enzyme. A method was developed utilizing an azo-dye-labelled collagen substrate that could measure the extracellular proteinase of serratia. The test was easy to perform, inexpensive, and potentially quantifiable. The azo dye test corresponded completely with the gelatinase tests. PMID- 2993835 TI - [Characteristics of protein synthesis in the West Nile virus and the nature of infection in mosquito and mammalian cells]. PMID- 2993837 TI - An examination of chronic angiotensin-converting enzyme inhibition in the rat. AB - We have examined the systemic and renal effects of 2 weeks' administration of angiotensin-converting enzyme inhibition (CEI) to both normal and chronic NaCl depleted Munich-Wistar rats and focused particularly on the factors contributing to the significant hypotension observed during surgery and anesthesia and the response of renal glomerular hemodynamics under these conditions. At renal micropuncture, mean arterial pressure was decreased in rats receiving CEI on normal and NaCl-depleted diets (72 +/- 5 and 78 +/- 6 mm Hg, p less than 0.01) but nephron filtration rate (sngfr) was well maintained since plasma flow was not altered and glomerular capillary pressure only slightly decreased. Negative Na+ balance in NaCl-depleted CEI rats correlated with decreased awake blood pressures. Under surgery, plasma norepinephrine and epinephrine were not increased in CEI rats in spite of hypotension, and diminished adrenergic responses may contribute to systemic and renal effects observed. Nephron plasma flow and sngfr are remarkably well preserved during CEI in spite of hypotension due to marked afferent arteriolar dilation. PMID- 2993836 TI - Urinary pCO2 as an index of collecting duct hydrogen ion secretion during chronic hypercapnia. AB - The rise in urinary pCO2 above blood pCO2 which occurs in response to bicarbonate loading (i.e. the urine to blood (U-B) pCO2 gradient), is used with increasing frequency as an index of collecting duct hydrogen ion secretion. We recently proposed, however, that the U-B pCO2 gradient is not an appropriate index of collecting duct hydrogen ion secretion when blood pCO2 is altered acutely. This issue was further investigated by examining the effect of chronic hypercapnia on urinary pCO2 generation. In rats exposed to chronic hypercapnia induced by breathing 10% CO2 for 3 days in an environmental chamber, acute sodium bicarbonate infusion resulted in a U-B pCO2 lower than that of normocapnic control rats (11 +/- 4.6 and 30 +/- 1.8 mm Hg, p less than 0.001). This finding could be interpreted to indicate that collecting duct hydrogen ion secretion is depressed in rats with chronic hypercapnia. The urinary pCO2 of rats with chronic hypercapnia was lower than that of the blood (54 +/- 6.0 and 86 +/- 1.2 mm Hg, p less than 0.005, respectively). In these rats, NaHCO3 infusion, while blood pCO2 was kept constant, elicited a marked rise in urine pCO2 (from 54 +/- 6.0 to 104 +/- 6.0 mm Hg, p less than 0.005) which was not significantly different from that observed in normocapnic control rats. The infusion of carbonic anhydrase resulted in a comparable fall in urine pCO2 in hypercapnic and normocapnic rats (-27 +/- 5 and -30 +/- 3 mm Hg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993838 TI - Nephrogenous cyclic AMP during extracellular volume expansion: relationship to early and late aortic constriction. AB - Prior data showed that both volume expansion (ECVE) and renal vasodilatation are associated with natriuresis and increased nephrogenous cAMP (NcAMP). Studies were done to examine the effect of ECVE on sodium excretion and NcAMP during early or late reduction in renal perfusion pressure (RPP) in an effort to define the role of renal vasodilatation on NcAMP during ECVE. In 8 thyroparathyroidectomized (TPTX) dogs, RPP to the left kidney was reduced (130 +/- 1.7 to 69 +/- 4 mm Hg), ECVE was then produced and finally RPP was allowed to rise to 126 +/- 2.2 mm Hg (group I). In 8 TPTX, ECVE was produced and then followed by a reduction in RPP (121 +/- 5 to 69 +/- 5 mm Hg; group II). cAMP was measured in aorta, renal veins and urine. In group I, renal plasma flow (RPF), fractional excretion of Na filtered (FeNa), urinary NcAMP (UNcAMP) and renal vein NcAMP (RVNcAMP) did not change in the constricted kidney while these parameters increased in the other kidney. After release of the constriction RPF (122 +/- 24 to 157 +/- 26 ml/min), FeNa (0.5 +/- 0.1 to 5 +/- 1.2%), UNcAMP (204 +/- 121 to 785 +/- 198 pmol/min, p less than 0.05) and RVNcAMP (277 +/- 213 to 1,565 +/- 403 pmol/min, p less than 0.02) rose significantly. In group II, FeNa, UNcAMP and RVNcAMP increased during ECVE and remained significantly elevated during the reduction in RPP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993839 TI - [Familial hypoglycocorticism syndrome unresponsive to ACTH, achalasia, alacrima, with associated distal neuromyopathy]. PMID- 2993840 TI - From ABO to HTLV. Changing trends in blood transfusion safety. PMID- 2993841 TI - Update: acquired immunodeficiency syndrome in the San Francisco Cohort Study, 1978-1985. PMID- 2993842 TI - Update: evaluation of human T-lymphotropic virus type III/lymphadenopathy associated virus infection in health-care personnel--United States. PMID- 2993843 TI - NIOSH Current Intelligence Bulletins: summaries. July 1985. PMID- 2993844 TI - Chlamydia trachomatis infections. Policy guidelines for prevention and control. PMID- 2993845 TI - [Management of malignant tumors of the thymus]. AB - Twenty-eight patients with malignant tumors of the thymus were surgically treated during the 15-year period from 1969 to 1984. Twenty-six patients had far advanced disease when treatment was began. Of fifteen patients with malignant thymoma, the tumors were successfully removed in five patients by using the technique of combined resection of SVC. Of two patients with carcinoid, one is alive and another died two years after extended operation. Of two patients with malignant lymphoma, one with non Hodgkin's disease died three years after operation, and the other with Hodgkin's disease is alive with tumor bearing. It is important to distinguish malignant lymphoma from other types of thymic tumors, because chemo radiotherapy is superior to surgical therapy. Of nine patients with germ cell tumor, two patients with seminoma have been well for 11 and 5 years. CDDP and radiotherapy were effective on their long survival. Three patients with embryonal carcinoma responded well to combined surgical and chemo-radiotherapy. But only one of them is alive with tumor free one year after operation. Prosthetic grafts were employed in 7 cases for replacement of the innominate and superior caval vein. Angiogram taken one month after operation disclosed the overall patency rate with 92% and the longest patent period confirmed was 4 years. Extended operation by using the prosthetic graft should be performed in the advanced cases to achieve a complete removal and to aim better prognosis for this kind of disease. In conclusion, aggressive surgical removal followed by radio chemotherapy offers best cure of the malignant thymic tumor. PMID- 2993846 TI - Homologous desensitization of the beta-adrenergic receptor. Functional integrity of the desensitized receptor from mammalian lung. AB - Previous work has demonstrated that injection of rats with isoproterenol is rapidly (10 min) followed by the development of a homologous form of desensitization of the beta-agonist-coupled adenylate cyclase in lung membranes. Half the receptor pool becomes sequestered in a light membrane fraction while the other half remains in the plasma membranes but becomes functionally uncoupled. In the present work we sought to assess whether "local sequestration" of the functionally intact receptor away from the effector adenylate cyclase in the plasma membrane contributes to the uncoupling of the beta-adrenergic receptor observed in the plasma membranes. We tested the functionality of the desensitized beta-adrenergic receptor in three different ways. We reconstituted the affinity chromatography purified control and "desensitized" receptors with pure Ns from human erythrocytes and assessed the ability to induce GTPase activity in Ns. Both control and desensitized beta-adrenergic receptors stimulate similar levels of GTPase activity in Ns (852 +/- 38 versus 738 +/- 49 fmol of Pi released/30 min (p greater than 0.05, n = 4). To further assess the relative ability of control and desensitized beta-adrenergic receptors to couple to another source of Ns we fused reconstituted beta-adrenergic receptors to Xenopus laevis erythrocytes, which contain Ns and adenylate cyclase but essentially no beta-adrenergic receptors. The functional interactions of control and desensitized beta-adrenergic receptor with the adenylate cyclase system of the acceptor cells was assessed by measuring the beta-agonist-stimulated adenylate cyclase activity and the agonist-induced formation of the high affinity state of the beta-adrenergic receptor (RH). Again both control and desensitized beta-adrenergic receptors appeared to interact with Ns to the same extent. To test if a local sequestration of the beta-adrenergic receptor away from Ns within the plasma membrane might contribute to the uncoupling of the beta-adrenergic receptors during desensitization, plasma membranes from control and desensitized lungs were treated with the fusogen polyethylene glycol to disrupt any compartmentalization of protein components within the plasma membrane. After polyethylene glycol treatment the previously uncoupled beta-adrenergic receptors could be recoupled to Ns as assessed by the formation of RH in agonist competition curves. These data suggest that in marked contrast to the heterologous type of desensitization, homologous desensitization may involve a local sequestration of a functionally intact beta-adrenergic receptor away from the adenylate cyclase effector system. PMID- 2993847 TI - Bicycloorthocarboxylate convulsants. Potent GABAA receptor antagonists. AB - 4-t-Butyl-1-(4-bromophenyl)-bicycloorthocarboxylate antagonizes gamma aminobutyric acid (GABA)-mediated relaxation at a functional insect nerve-muscle synapse, mimicking the action of picrotoxinin, suggesting that it causes GABA antagonism through blockade of the chloride ionophore. It is also a potent GABAA receptor antagonist, inhibiting the binding of [35S]t-butyl bicyclophosphorothionate ([35S]TBPS) to EDTA/water-dialyzed human brain P2 membranes. Structure-activity relationships of 74 1,4-bis-substituted bicycloorthocarboxylates, mostly new compounds, reveal that for high potency as a GABAA receptor antagonist the optimal 4-substituent is a C4 to C6 branched chain alkyl or cycloalkyl group (e.g., t-butyl, s-butyl, or cyclohexyl) and the optimal 1-substituent is a phenyl moiety with one or more electron-withdrawing groups (e.g., 4-cyano, 4-bromo, 4-chloro, 3,4-dichloro, or pentafluoro). Bicycloorthocarboxylate inhibitors of [35S]TBPS binding with IC50 values of 5-10 nM exceed by several-fold the potency of any GABAA receptor antagonist previously reported. The 4-t-butyl-1-(4-azidophenyl) analog, synthesized as a candidate photoaffinity label, gives an IC50 of 315 nM. The potency of bicycloorthocarboxylates for decreasing [35S]TBPS binding generally correlates with their toxicity, i.e., compounds without inhibitory activity in this brain receptor assay are of low toxicity on intraperitoneal administration to mice, and the analogs most potent as inhibitors are generally those most toxic to mice (e.g., IC50 of 5 nM and LD50 of 0.06 mg/kg for 4-t-butyl-1-(4-cyanophenyl) bicycloorthocarboxylate). The effects of phenyl substituents on the potency of the orthobenzoates as GABAA receptor antagonists are similar to those on toxicity. In contrast to the 1-substituted phenyl compounds, 4-t-butyl-1-ethynyl bicycloorthocarboxylate and its 4-i-propyl analog are very toxic (LD50 0.4-2 mg/kg) but have only moderate inhibitory potency (IC50 480-2900 nM), a pattern noted for many 1-alkyl-bicycloorthocarboxylates, suggesting that even within this series there may be different types of receptor-inhibitor interactions. 1-(4 Chlorophenyl)-4-cyclohexyl-bicycloorthocarboxylate is particularly sensitive to oxidative detoxification based on its 10-fold synergism of toxicity by piperonyl butoxide and marked potency loss in a coupled [35S]TBPS receptor/microsomal oxidase assay. Some benzodiazepines and phenobarbital protect against poisoning by 1-(4-bromophenyl)- and 1-ethynyl-4-t-butyl-bicycloorthocarboxylates and their 1-(4-bromophenyl)-4-cyclohexyl analog.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993848 TI - Phenyliminoimidazolidines. Characterization of a class of potent agonists of octopamine-sensitive adenylate cyclase and their use in understanding the pharmacology of octopamine receptors. AB - Octopamine, a major aminergic neurotransmitter in invertebrates, exerts many of its actions through receptors which are associated with the activation of adenylate cyclase. The present study defines and characterizes a new class of potent octopamine agonists, the substituted phenyliminoimidazolidines (PIIs). Approximately 30 of these derivatives were examined for agonist and antagonist effects on the highly enriched and specific octopamine-sensitive adenylate cyclase present in the firefly light organ, as well as on adenylate cyclases present in other invertebrate and vertebrate tissues. Several derivatives were extremely active and some (e.g. 2,6-diethyl-PII) had potencies exceeding those of any previously described agonists of octopamine-sensitive adenylate cyclase. Stimulation by the potent PIIs was reversible, nonadditive to that caused by octopamine, and could be antagonized by antagonists such as cyproheptadine (Ki = 4 microM), phentolamine (Ki = 23 microM), and propranolol (Ki = 72 microM). These inhibitory constants agreed well with those for inhibiting octopamine stimulation. Certain PII derivatives acted as partial agonists and some as antagonists of octopamine stimulation. Structure-activity relationships revealed, among other things, that short-chain alkyl substitution in the 2- and 6-phenyl positions enhanced activity, as did further substitution of 4-halo, 4-methyl, or 4-hydroxy substituents. 4-Amino or N-alkyl substitution decreased activity. Structurally related benzylimidazoline derivatives such as tolazoline and naphazoline were partial octopamine agonists, generally less active than the PIIs. Comparison, in three invertebrate species, of the effects of the PIIs and two other chemical classes of octopamine agonists demonstrated clearcut differences in species responsiveness. Other comparative studies revealed that the agonist activity of the potent PIIs was specific for tissues containing an octopamine-sensitive adenylate cyclase; adenylate cyclases activated by dopamine or by beta 1- or beta 2-adrenergic agonists were unaffected by these compounds. Evaluation of the relative binding affinities of various PIIs for mammalian alpha adrenergic receptors, as well as the ability of various antagonists to block PII binding, strongly suggested that the active PIIs are affecting a class of octopamine receptors distinct from mammalian alpha 1- or alpha 2-adrenergic receptors. These octopamine receptors also appeared distinct from mammalian 5-HT1 and 5-HT2 receptors. Correlative physiological studies in insects revealed that the active PIIs mimicked octopamine and were potent activators of light emission in the firefly light organ.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 2993849 TI - The accessibility of phage MS2 RNA to structure specific nucleases in various conditions. AB - The accessibility of ds- and ss-segments of phage MS2 RNA to ds- and ss-specific nucleases (RNase III, nuclease SV and nuclease S1) was studied. The results show that the RNA has hydrolysis sites for all the nucleases used. These sites are unvariable in a wide range of the conditions (ionic strength, pH, bivalent cations and temperature) and are not changed also after denaturation-renaturation of the RNA. This testifies that the distribution and interactions of ds- and ss segments in the whole molecule are very specific and stable. PMID- 2993850 TI - A site-specific endonuclease from Pseudomonas aeruginosa. AB - PaeI, a new restriction endonuclease from Pseudomonas aeruginosa clinical strain was isolated and characterized. It recognizes and cleaves the sequence 5'-GCATG reduced C-3' generating DNA fragments with 3'-tetranucleotide sticky ends. DNAs of pBR322, SV40 and bacteriophage lambda have one, two and six PaeI recognition sites, respectively. Seventy-two strains of Pseudomonas, Clostridium, Escherichia coli, Shigella, Proteus and Saccharomyces were screened for the presence of site specific endonucleases. Here we describe the PaeI restriction enzyme found in Pseudomonas aeruginosa; other data will be published elsewhere. Earlier Hinkle and Miller isolated from P. aeruginosa a PaeR7 restriction endonuclease recognizing and cleaving a sequence 5'-C reduced TCGAG-3' (1). Sequence analysis of DNAs cleaved by PaeI shows that the enzyme is the isoschizomer of SphI (2). PMID- 2993852 TI - [Lambda plasmidophages and their properties]. AB - Plasmidphage lambda NM::pBR322 has been constructed in vitro and characterized. Under normal conditions the hybrid DNA molecule undergoes a lytic cycle of phage development, whereas in the presence of antibiotic lambda DNA replicates in the cell extrachromosomally as a plasmid. Properties of plasmidphage lambda NM::pBR322 have been compared with the earlier constructed lambda gt::pMB9. It has been demonstrated that plasmid pMB9 in vivo can be precisely excised from the lambda gt::pMB9. PMID- 2993851 TI - Processive cleavage of concatemer DNA duplexes by Eco RII restriction endonuclease. AB - Eco RII restriction endonuclease cleaves synthetic DNA-duplexes in which the recognition sites of this enzyme (5'...CCATGG...) are repeated every 9 base pairs with the alternating orientation of the central AT pair. It operates in a processive mode, i.e. the bound enzyme molecule slides along the substrate toward neighboring recognition sites. Nona-nucleotides are the main products of the cleavage. The data obtained neighboring recognition sites. Nona-nucleotides are the main products of the cleavage. The data obtained point to the capability of Eco RII endonuclease to recognize and cleave the substrate under both possible orientations of the central AT-pair of the recognition site with respect to the bound enzyme molecule. These data also show the close similarity of DNA structures in a complex with the enzyme and without. PMID- 2993853 TI - [Kinetic model of the effect of dehydration on electron transfer from membrane bound cytochrome c to the photosynthetic reaction center]. AB - Amplitude characteristics and kinetics of laser-induced oxidation of high potential cytochrome CH by a photosynthetic reaction center (RC) were investigated in Ectothiorhodospira shaposhnikovii chromatophore preparations of various humidity. It is shown that the diminuition of the amount of oxidized cytochrome and the decrease of the rate of the reaction on lowering the preparation humidity can be explained in terms of the concept of conformation controlled electron transfer within the CH-RC complex. A model is suggested which predicts that the reversible transition of the complex from one conformational state which allows electron transfer ("contact" state) to the other in which the transfer is impossible ("non-contact" state) is the result of drying (or low temperature) induced changes in the electron tunnelling path in the region of "contact" of the cytochrome CH and RC protein globules. PMID- 2993854 TI - [Introduction of DNA sequences of Rous sarcoma virus into Drosophila and mouse genomes by microinjections into ova]. AB - The paper covers experimental results of introducing exogenic genetic material, namely DNA sequences of the Rous sarcoma virus, by microinjections in mice zygotes and Drosophila early embryos. In a number of cases integration of viral DNA into genomes of these organisms was detected. Blot-hybridizations analysis of cell DNA proved that the inserted viral sequences undergo rearrangements in the course of integration. PMID- 2993855 TI - [Determinant of resistance to kanamycin in Streptomycin rimosus: amplification in the chromosome and reversible genetic instability]. AB - The study on the kanamycin resistance determinant (Kanr) in an oxytetracycline- producing strain of S. rimosus showed that it was capable of amplifying in the chromosome during selection for increasing the antibiotic resistance level. The amplification of the DNA fragment with a molecular weight of 10.3 MDa containing Kanr amounted to 300 copies per genome, which resulted in a more than 1000-fold increase in kanamycin resistance level. Cloning of the Kanr determinant on plasmid SLP1.2 in S. lividans strain 66 was performed. In Streptomyces lividans strain 66 the Kanr determinant preserved the capacity for amplification in the hybrid plasmid pSU10 integrated into the chromosome. The Kanr determinant in the strains of S. rimosus and S. lividans was characterized by transfers Kanr in equilibrium Kans with a frequency of 1 X 10(-3). It was shown that the mutation in S. lividans strain 66 resulting in phenotype Kans was not connected with the structural Kanr gene on plasmid pSU10 but was localized on the chromosome. Phenotype Kans was promoted by a decrease in the number of the copies of the regulatory genetic element designated RES1. The reverse to phenotype Kanr might be due to one of the following events: amplification to the initial level of RES1 and amplification up to 200 copies per the genome of the hybrid plasmid pSU10 containing the Kanr determinant. Amplification of the Kanr determinant with preserved initial level of RES1 element resulted in a more than 1000 times increase in the resistance level. PMID- 2993856 TI - [The region of phage T4 W-29 genes: cloning and expression]. AB - The EcoRI fragment of T4 DNA containing the W-29 genes and its subfragments were cloned in the pBR322 and the singlestranded M13 phage. Hybridization with a cloned DNA showed that in T4 infected cells the transcription of the late genes 25-29 depends on the phage-induced RNA polymerase changes and on replication of phage DNA. At a late infection stage one also observes an enhanced transcription of the (early) genes uvsW and uvsY, which depends on viral DNA replication. Both early and late genes within recombinant plasmids are also expressed in uninfected cells carrying a plasmid regardless of the inserted fragment orientation and independently of the vector promoters. Hybrid plasmids demonstrated a high frequency of recombination with phage DNA in the infected cell. An RNA polymerase from uninfected cells binds itself to the late cloned genes to form "open" complexes. A purified RNA polymerase transcribes both early and late genes within recombinant plasmids. The relative transcription of the late cloned genes is enhanced if one uses an RNA polymerase from T4-infected cells. The super-helicity of template DNA is essential for transcription of early and late genes. PMID- 2993857 TI - Control of carbohydrate processing: the lec1A CHO mutation results in partial loss of N-acetylglucosaminyltransferase I activity. AB - Lec1 CHO cell glycosylation mutants are defective in N acetylglucosaminyltransferase I (GlcNAc-TI) activity and therefore cannot convert the oligomannosyl intermediate (Man5GlcNAc2Asn) into complex carbohydrates. Lec1A CHO cell mutants have been shown to belong to the same genetic complementation group but exhibit different phenotypic properties. Evidence is presented that lec1A represents a new mutation at the lec1 locus resulting in partial loss of GlcNAc-TI activity. Structural studies of the carbohydrates associated with vesicular stomatitis virus grown in Lec1A cells (Lec1A/VSV) revealed the presence of biantennary and branched complex carbohydrates as well as the processing intermediate Man5GlcNAc2Asn. By contrast, the glycopeptides from virus grown in CHO cells (CHO/VSV) possessed only fully processed complex carbohydrates, whereas those from Lec1/VSV were almost solely of the Man5GlcNAc2Asn intermediate type. Therefore, the Lec1A glycosylation phenotype appears to result from the partial processing of N-linked carbohydrates because of reduced GlcNAc-TI action on membrane glycoproteins. Genetic experiments provided evidence that lec1A is a single mutation affecting GlcNAc-TI activity. Lec1A mutants could be isolated at frequencies of 10(-5) to 10(-6) from unmutagenized CHO cell populations by single step selection, a rate inconsistent with two mutations. In addition, segregants selected from Lec1A X parental cell hybrid populations expressed only Lec1A or related lectin-resistant phenotypes and did not include any with a Lec1 phenotype. The Lec1A mutant should be of interest for studies on the mechanisms that control carbohydrate processing in animal cells and the effects of reduced GlcNAc-TI activity on the glycosylation, translocation, and compartmentalization of cellular glycoproteins. PMID- 2993858 TI - Simian virus 40 DNA replication in vitro: specificity of initiation and evidence for bidirectional replication. AB - We recently described a soluble cell-free system derived from monkey cells that is capable of replicating exogenous plasmid DNA molecules containing the simian virus 40 (SV40) origin of replication (J.J. Li, and T.J. Kelly, Proc. Natl. Acad. Sci. U.S.A. 81:6973-6977, 1984). Replication in the system is completely dependent upon the addition of the SV40 large T antigen. In this report we describe additional properties of the in vitro replication reaction. Extracts prepared from cells of several nonsimian species were tested for the ability to support origin-dependent replication in the presence of T antigen. The activities of extracts derived from human cell lines HeLa and 293 were approximately the same as those of monkey cell extracts. Chinese hamster ovary cell extracts also supported SV40 DNA replication in vitro, but the extent of replication was approximately 1% of that observed with human or monkey cell extracts. No replication activity was detectable in extracts derived from BALB/3T3 mouse cells. The ability of these extracts to support replication in vitro closely parallels the ability of the same cells to support replication in vivo. We also examined the ability of various DNA molecules containing sequences homologous to the SV40 origin to serve as templates in the cell-free system. Plasmids containing the origins of human papovaviruses BKV and JCV replicated with an efficiency 10 to 20% of that of plasmids containing the SV40 origin. Plasmids containing Alu repeat sequences (BLUR8) did not support detectable DNA replication in vitro. Circular DNA molecules were found to be the best templates for DNA replication in the cell-free system; however, linear DNA molecules containing the SV40 origin also replicated to a significant extent (10 to 20% of circular molecules). Finally, electron microscopy of replication intermediates demonstrated that the initiation of DNA synthesis in vivo takes place at a unique site corresponding to the in vivo origin and that replication is bidirectional. These findings provide further evidence that replication in the cell-free system faithfully mimics SV40 DNA replication in vivo. PMID- 2993859 TI - Recombination between poly[d(GT).d(CA)] sequences in simian virus 40-infected cultured cells. AB - CVI cells were transfected with oversized simian virus 40 (SV40) genomes that could be reduced to packageable size by alternative homologous recombination pathways involving either two polydeoxyguanylic-thymidylic acid X polydeoxycytidylic-adenylic acid (poly[d(GT).d(CA)]; abbreviated hereafter as poly(GT)] tracts or two tracts of homologous SV40 sequence. Plaque-forming viruses rescued by this procedure were found to contain genomes formed by homologous and nonhomologous recombination events. Half of the viable viral DNA molecules recovered were the result of recombination between two tracts of poly(GT). Approximately 20% of the rescued viral genomes were produced by homologous recombination between tracts of SV40 DNA. Nonhomologous recombination involving SV40 sequences was also a major pathway of deletion, producing ca. 30% of the viral plaques. Tracts of poly(GT) generated by recombination were variable in length, suggesting that recombination between poly(GT) tracts was usually unequal. On a per-nucleotide basis, poly(GT) recombination occurred eight times more frequently than did recombination between homologous SV40 DNA. This eightfold difference is the maximum recombinatory enhancement attributable to poly(GT) sequences. Although DNA sequence analysis showed that tracts of poly(GT) generated by recombination retained the alternating G-T repeat motif throughout their length, the contribution of the nonhomologous pathway to poly(GT) recombination cannot be ruled out, and the relative proclivity of a given length of d(GT).d(CA) sequence to undergo homologous recombination is probably less than eight times greater than that of an SV40 sequence of the same length. PMID- 2993860 TI - Attenuation of late simian virus 40 mRNA synthesis is enhanced by the agnoprotein and is temporally regulated in isolated nuclear systems. AB - Studies were performed to verify the physiological significance of attenuation in the life cycle of simian virus 40 and the role of agnoprotein in this process. For these purposes, nuclei were isolated at various times after infection and incubated in vitro in the presence of [alpha-32P]UTP under the standard conditions which lead to attenuation. Attenuation was evident by the production of a 94-nucleotide attenuator RNA, revealed by gel electrophoresis. In parallel, the synthesis of agnoprotein was studied at various times after infection by labeling the cells for 3 h with [14C]arginine, lysing them, and analyzing the labeled proteins by gel electrophoresis. Both attenuation and the synthesis of agnoprotein were predominant towards the end of the infectious cycle. At earlier times, there was almost no attenuation and no synthesis of agnoprotein. Moreover, there was almost no attenuation even at the latest times after infection in nuclei isolated from cells infected with simian virus 40 deletion mutants that do not synthesize agnoprotein. Finally, analysis by dot blot hybridization showed higher amounts of cytoplasmic viral RNA in cells infected with an agnoprotein gene insertion mutant, delta 79, that does not produce agnoprotein, compared with cells infected with wild-type virus. The present studies indicate that attenuation is temporally regulated and suggest that agnoprotein enhances attenuation in isolated nuclei and that may also enhance it in vivo. PMID- 2993861 TI - H-2Ld antigen encoded by a recombinant retrovirus genome is expressed on the surface of infected cells. AB - A recombinant murine retrovirus was constructed which contains, within its genome, a truncated version of the gene encoding the murine H-2Ld major histocompatibility antigen. The H-2Ld gene, which was inserted 3' of the env splice acceptor site in the recombinant retrovirus MSV-neo, lacked the 5' promoter and TATA sequences and the 3' transcription termination and polyadenylate addition sites of the normal H-2Ld gene. Transfection of the MSV neo/H-2Ld plasmid (pLTV-11) into Y-2 cells resulted in the production of the transmissible recombinant retrovirus LTV-11. Cells infected with LTV-11 virus were resistant to the eucaryotic antibiotic G418 and expressed H-2Ld on the cell surface. These infected cells contained a viral RNA species which possessed both the H-2Ld and the neomycin resistance gene sequences but did not contain significant levels of the smaller H-2Ld-specific mRNA. The H-2Ld antigen expressed on the surface of infected cells functioned as a target for cytolytic T cells specific for the H-2Ld antigen. PMID- 2993862 TI - trans Activation of the simian virus 40 late transcription unit by T-antigen. AB - We have investigated the role of simian virus 40 (SV40) T-antigen in the induction of late gene expression independent of its function in amplifying templates through DNA replication. Northern blot and S1 nuclease analyses showed that stimulation occurred at the transcriptional level. At least two template elements, the T-antigen-binding sites and the 72-base-pair repeats, appeared to be important for this induction. Using template mutants, we demonstrated that deletions within T-antigen-binding site II decreased T-antigen-mediated late gene expression approximately 10- to 20-fold. In addition, multiple point mutations within a single retained copy of the SV40 72-base-pair repeat decreased T-antigen mediated late gene expression. Using in vivo competition studies, we demonstrated that competitor DNA fragments containing the SV40 control region (nucleotides 5171 through 272) quantitatively decreased SV40 late gene expression in COS-1 cells. In contrast, competition with a plasmid containing SV40 nucleotides 1 through 294 (which removes all of T-antigen-binding site I and half of site II) was much less efficient. Finally, we demonstrated that in vivo competition experiments employing competitor fragments distal to the T-antigen-binding sites within the late template region (SV40 nucleotides 180 through 2533) resulted in superinduction of late gene expression in COS-1 cells. This finding suggests that negative factors such as repressors or attenuators may modulate late SV40 gene expression before induction. Our results are consistent with a model in which induction of late gene expression involves an interaction of the SV40 origin region with DNA-binding proteins, one of which may be T-antigen. Activation of the SV40 late transcription unit may involve induction of the SV40 enhancer or removal of a repressor-like protein or both. PMID- 2993864 TI - Incorporation of a charged amino acid into the membrane-spanning domain blocks cell surface transport but not membrane anchoring of a viral glycoprotein. AB - The membrane-spanning domain of the vesicular stomatitis virus glycoprotein (G protein) consists of a continuous stretch of 20 uncharged and mostly hydrophobic amino acids. We examined the effects of two mutations which change the amino acid sequence in this domain. These mutations were generated by oligonucleotide directed mutagenesis of a cDNA clone encoding the G protein, and the altered G proteins were then expressed in animal cells. Replacement of an isoleucine residue in the center of this domain with a strongly polar but uncharged amino acid (glutamine) had no effect on membrane anchoring or transport of the protein to the cell surface. Replacement of this same isoleucine residue with a charged amino acid (arginine) generated a G protein that still spanned intracellular membranes but was not transported efficiently to the cell surface. The protein accumulated in the Golgi region in about 50% of the cells, and about 20% of the cells had detectable protein levels in a punctate pattern on the cell surface. In the remaining cells the protein accumulated in a vesicular pattern throughout the cytoplasm. Models which might explain the abnormal behavior of this protein are discussed. PMID- 2993863 TI - Structure and biological activity of human homologs of the raf/mil oncogene. AB - Two human genes homologous to the raf/mil oncogene have been cloned and sequenced. One, c-raf-2, is a processed pseudogene; the other, c-raf-1, contains nine exons homologous to both raf and mil and two additional exons homologous to mil. A 3' portion of c-raf-1 containing six of the seven amino acid differences relative to murine v-raf can substitute for the 3' portion of v-raf in a transformation assay. Sequence homologies between c-raf-1 and Moloney leukemia virus at both ends of v-raf indicate that the viral gene was acquired by homologous recombination. Although the data are consistent with the traditional model of retroviral transduction, they also raise the possibility that the transduction occurred in a double crossover event between proviral DNA and the murine gene. PMID- 2993865 TI - Unstable expression and amplification of a transfected oncogene in confluent and subconfluent cells. AB - NIH 3T3 cells were transfected with a plasmid containing the transforming gene, v src, from Rous sarcoma virus. One of the transformed cell lines isolated reverted to a flat, nontransformed morphology after cloning through soft agar. This cell line did not express the src gene and could no longer grow in soft agar. When these cells were held at confluence, spontaneous foci appeared which eventually covered the dish. The appearance of foci correlated with an increase in v-src gene expression, ability to grow in soft agar, and tumorigenicity in mice. When these transformed cells were trypsinized and held at subconfluence, both v-src expression and the transformed phenotype were progressively lost. Whereas rearrangement of the transfected gene was not detected, the gene copy number in the transformed cells was markedly increased (greater than 50-fold). Confluence dependent gene amplification and deamplification have been retained after several cycles of growth alternately at high and low density, in cells recloned through soft agar, and after cells had been maintained continuously at high or low density. The results suggest that, in this cell line, reversible gene amplification plays a central role in expression of the transfected gene. PMID- 2993866 TI - Building a metal-responsive promoter with synthetic regulatory elements. AB - A fusion gene consisting of the promoter region from the mouse metallothionein-I gene joined to the coding region of the herpes simplex virus thymidine kinase gene is efficiently regulated by zinc in a transient assay when transfected into baby hamster kidney cells. Analysis of similar plasmids in which the metallothionein-I promoter region was mutated indicated the presence of multiple metal regulatory elements (MREs) between -176 and -44 base pairs from the cap site. To further investigate the function of MREs, we inserted a synthetic DNA fragment containing the sequence of MRE-a (the element between -55 and -44 base pairs) into the nonresponsive promoter of the thymidine kinase gene in various positions and configurations. Little or no induction by zinc was observed with single insertions of the regulatory sequence, whereas many different constructions having two copies of MRE-a were inducible. The precise position of the two MREs relative to each other or to the thymidine kinase promoter elements had a relatively small effect on the efficiency of induction, but the inducibility could be further increased by the introduction of more MRE-a sequences. MRE-a can function synergistically with the thymidine kinase distal promoter elements, but in the presence of the TATA box alone it functions as a positive, zinc-dependent promoter element. PMID- 2993867 TI - Induction of cellular thymidine kinase occurs at the mRNA level. AB - The thymidine kinase (TK) gene has been isolated from human genomic DNA. The gene was passaged twice by transfection of LTK- cells with human chromosomal DNA, and genomic libraries were made in lambda Charon 30 from a second-round TK+ transformant. When the library was screened with a human Alu probe, seven overlapping lambda clones from the human TK locus were obtained. None of the seven contained a functional TK gene as judged by transfection analysis, but several combinations of clones gave rise to TK+ colonies when cotransfected into TK- cells. A functional cDNA clone encoding the human TK gene has also been isolated. Using this cDNA clone as a probe in restriction enzyme/blot hybridization analyses, we have mapped the coding sequences and direction of transcription of the gene. We have also used a single-copy subclone from within the coding region to monitor steady-state levels of TK mRNA in serum-stimulated and simian virus 40-infected simian CV1 tissue culture cells. Our results indicate that the previously reported increase in TK enzyme levels seen after either treatment is paralleled by an equivalent increase in the steady-state levels of TK mRNA. In the case of simian virus 40-infected cells, the induction was delayed by 8 to 12 h, which is the length of time after infection required for early viral protein synthesis. In both cases, induction of TK mRNA coincides with the onset of DNA synthesis, but virally infected cells ultimately accumulate more TK mRNA than do serum-stimulated cells. PMID- 2993868 TI - Sequences in human cytomegalovirus which hybridize with the avian retrovirus oncogene v-myc are G + C rich and do not hybridize with the human c-myc gene. AB - The degree of relatedness between previously identified cross-hybridizing regions within human cytomegalovirus strain AD169 and the avian retrovirus oncogene v-myc were investigated by nucleotide sequence comparison. We found that the homologous regions between the human cytomegalovirus genome and v-myc are limited to short G + C-rich regions in each genome and that the human cytomegalovirus genome shares little or no homology with the human c-myc gene. PMID- 2993869 TI - Induction of cellular DNA synthesis by a simian virus 40 mutant defective in nuclear transport of T antigen. AB - The simian virus 40 (SV40) (cT)-3 mutant [SV40(cT)-3], which is defective in nuclear transport of T antigen, was utilized to determine whether cellular DNA synthesis can be stimulated by SV40 in the absence of detectable nuclear T antigen. Cellular DNA synthesis was examined in the temperature-sensitive cell cycle mutants, BHK ts13 and BHK tsAF8, after microinjection of quiescent cells with plasmid DNA containing cloned copies of wild-type SV40 or SV40(cT)-3. The efficiency of induction of cellular DNA synthesis was identical for both wild type SV40 and SV40(cT)-3 in both cell lines. The results suggest that cell surface-associated T antigen, either alone or possibly in combination with minimal amounts of nuclear T antigen below our limit of detection, is able to stimulate cellular DNA synthesis. PMID- 2993870 TI - Host species specificity of polyomavirus DNA replication is not altered by simian virus 40 72-base-pair repeats. AB - The simian virus 40 72-base-pair repeats substituted for the polyomavirus enhancer, allowing replication and transcription in mouse 3T6 but not monkey CV-1 cells. A polyomavirus genome containing the entire simian virus 40 control region replicated at low levels in 3T6 and CV-1 cells; however, transcripts were detected only in 3T6 cells. Our results suggest that the simian virus 40 72-base pair repeats are unable to alter the host species specificity of the complete polyomavirus genome. PMID- 2993871 TI - High-resolution mapping of S1- and DNase I-hypersensitive sites in chromatin. AB - A new and easy technique for accurately mapping DNase I- and S1 nuclease hypersensitive sites is described. The technique is a modification of primer extension and S1 nuclease methods conventionally used to map RNA ends. PMID- 2993872 TI - Nature of the interaction between the C1q and C1r2S2 subunits of the first component of human complement. AB - The strength of interaction between the C1q and C1r2S2 subunits of C1 was studied as a function of temp. During centrifugation through sucrose density gradients at 4 degrees C, macromolecular C1 readily dissociated as it sedimented away from its free subunits. In contrast, at 20 degrees C, C1 remained associated as the 16S complex throughout centrifugation, thus indicating a stronger interaction between C1q and C1r2S2 at the higher temp. C1-inhibitor (C1-In) or nitrophenylguanidinobenzoate was present during centrifugation to prevent C1 activation. That native C1 was in fact the species being studied was confirmed by SDS-PAGE analysis. To investigate this temp dependence without using inhibitors, an alternative approach was used. Trace amounts of 125I-C1q were centrifuged through numerous sucrose density gradients, each of which contained a different concn of native C1r2S2 throughout the gradient. The s-rate of 125I-C1q increased with increasing C1r2S2 input. An association constant of 4.9 X 10(7) M-1 was calculated for this reversible interaction at 4 degrees C. However, at 20 degrees C, the data indicated a much higher affinity reaction since the addition of far less C1r2S2 was required for the s-rate of 125I-C1q to reach the 16S plateau. The presence of Cl-In did not affect these results. We have demonstrated that the association of C1q with C1r2S2 increases with increasing temp, a finding suggestive of a hydrophobic interaction. However, since we also show that C1 readily dissociates with increasing NaCl concn, the C1q-C1r2S2 interaction must, in fact, be ionic in nature. We therefore conclude that the temp dependence of the inter-subunit interaction is the result of a conformational change(s) within one of the subunits, and propose that this change may be similar to that occurring during Cl activation. PMID- 2993873 TI - Natural killer cells inhibit outgrowth of autologous Epstein-Barr virus-infected B lymphocytes. AB - To determine whether natural killer (NK) cells are the cells responsible for inhibition of outgrowth of Epstein-Barr virus (EBV)-infected autologous B lymphocytes, NK-enriched or NK-depleted populations were prepared by Percoll density gradient fractionation and complement lysis depletion of cells reacting with NK-specific monoclonal antibody HNK-1. These cells were then examined in parallel for NK activity and inhibition of outgrowth. NK-enriched low density cells inhibited outgrowth whereas NK-depleted high density cells did not. Low density cells treated with monoclonal antibodies HNK-1 and DR plus complement had little NK activity and failed to inhibit EBV-induced outgrowth, whereas these same cells treated with monoclonal antibodies OKT3 and DR plus complement had strong NK activity and caused marked inhibition of outgrowth. These findings indicate that NK cells rather than mature T cells, monocytes, or B cells, are responsible for inhibition of EBV-induced B cell outgrowth. PMID- 2993874 TI - Induction of sister-chromatid exchanges by restriction endonucleases. AB - Restriction endonucleases Cfo 1, Pvu II, Sma I, Hpa II, Taq I and Hae III were tested for their ability to induce SCEs in CHO cells. The results indicate that the DNA double-strand breaks induced during S-phase by these enzymes lead to an increase in the frequencies of SCEs. PMID- 2993875 TI - Heat-shock-induced enhanced reactivation of UV-irradiated Herpesvirus. AB - The objective of this study was to compare the ability of heat shock (HS) with that of another type of cellular stress, UV irradiation, to cause the induction of enhanced viral reactivation, a process that may represent an SOS-type repair process in mammalian cells. Studies performed to evaluate the effect of HS on growth of Vero cells revealed that HS at 45 degrees C for 45 min caused inhibition of cell growth similar to that caused by UV irradiation at 12 J/m2, but this inhibition was not observed at HS treatment for 5-15 min, or at a UV fluence of 2 J/m2. Enhanced reactivation of UV-irradiated Herpesvirus was observed in cells which had been pretreated by HS for greater than 30 min or UV at 12 J/m2. The synthesis of new proteins following HS for 15 and 45 min and UV at 12 J/m2 was examined by [35S]methionine-labeling experiments. The new synthesis of two HS proteins with molecular weights of 46 000 and 78 000 was induced by both levels of HS, but to a much greater extent at the high dose. These proteins were not detected in response to UV irradiation. These results indicate that, like UV irradiation, HS at levels inhibitory to cell growth induced enhanced viral reactivation in Vero cells. The results also suggest that at least two proteins in the HS protein family are not necessary for this response to occur. PMID- 2993876 TI - Uvr-independent repair of 8-methoxypsoralen crosslinks in Escherichia coli: evidence for a recombinational process. AB - On the basis of survival data, repair of 8-methoxypsoralen DNA crosslinks in Escherichia coli strains lacking a functional uvrABC endonuclease, is shown to require the products of the recA, recB, recF and recN genes. Bacteria, grown under conditions where most cells contain only a single genome, show no evidence of crosslink repair. Similarly, bacteriophage lambda shows evidence of crosslink repair only in SOS-induced cells, and only at multiplicities of infection greater than 1. The requirement for rec+ genes may be partly ascribed to the need for a functional SOS response, but taken together, the results suggest a recombinational step involving a homologous region of DNA may occur during uvr independent crosslink repair. PMID- 2993877 TI - Ability of various alkylating agents to induce adaptive and SOS responses: a study with lacZ fusion. AB - We used alkA'-lacZ' and umuC'-lacZ' fused genes and determined the ability of various alkylating agents to induce adaptive and SOS responses. The degree of induction of expression of these genes was quantitatively measured by a simple colorimetric assay of beta-galactosidase activity. SN1 type methylating agents, such as N-methyl-N'-nitro-N-nitrosoguanidine and N-methyl-N-nitrosourea, were more effective inducers for the alkA than for the umuC system, while SN1 type ethylating agents, such as N-ethyl-N'-nitro-N-nitrosoguanidine and N-ethyl-N nitrosourea, were more potent inducers for the umuC than for the alkA system. Similar but less striking effects on the two systems were obtained with SN2 type alkylating agents. PMID- 2993878 TI - A minor pathway of postreplication repair in Escherichia coli is independent of the recB, recC and recF genes. AB - After ultraviolet (UV) irradiation, an Escherichia coli K12 uvrB5 recB21 recF143 strain (SR1203) was able to perform a limited amount of postreplication repair when incubated in minimal growth medium (MM), but not if incubated in a rich growth medium. Similarly, this strain showed a higher survival after UV irradiation if plated on MM versus rich growth medium (i.e., it showed minimal medium recovery (MMR]. In fact, its survival after UV irradiation on rich growth medium was similar to that of a uvrB5 recA56 strain, which does not show MMR or postreplication repair. The results obtained with a uvrB5 recF332::Tn3 delta recBC strain and a uvrB5 recF332::Tn3 recB21 recC22 strain were similar to those obtained for strain SR1203, suggesting that the recB21 and recF143 alleles are not leaky in strain SR1203. The treatment of UV-irradiated uvrB5 recB21 recF143 and uvrB5 recF332::Tn3 delta recBC cells with rifampicin for 2 h had no effect on survival or the repair of DNA daughter-strand gaps. Therefore, a pathway of postreplication repair has been demonstrated that is constitutive in nature, is inhibited by postirradiation incubation in rich growth medium, and does not require the recB, recC and recF gene products, which control the major pathways of postreplication repair. PMID- 2993879 TI - Sphingomyelin synthesis in Hymenolepis diminuta (Cestoda). AB - Adult Hymenolepis diminuta incorporated label from L[U-14C]serine, [1 14C]palmitic acid, [1-14C]palmitoyl-CoA and cytidine-5'-diphospho[methyl 14C]choline into the various intermediates of sphingomyelin synthesis (ketosphingosine, dihydrosphingosine, sphingosine, ceramide and sphingomyelin). From the results it was concluded that H. diminuta possessed the five enzymes involved in sphingomyelin synthesis, namely serine palmitoyl-transferase, 3 oxosphinganine reductase, flavoprotein dihydrosphingosine reductase, sphingosine acyltransferase and ceramide choline-phosphotransferase. PMID- 2993880 TI - The two mechanisms for antigenic variation in Trypanosoma brucei are independent processes. AB - Antigenic switching in Trypanosoma brucei can occur either by the production of a telomeric copy of a variant surface glycoprotein (VSG) gene through a gene conversion mechanism or by the nonduplicative activation of a telomeric VSG gene. The 5 VSG gene telomeric copy that is expressed in IsTaR 1 variant antigenic type (VAT) 5 is retained in an inactive state following an antigenic switch to VAT A5. This inactive telomeric 5 VSG gene copy is absent following independent single antigenic switches to VATs 1A5 and 11A5. The inactive 5 VSG gene does not appear to have been replaced with the newly expressed VSG gene. Thus, inactive telomeric VSG genes that are capable of being expressed can be lost, presumably through gene conversion to new VSG genes. These results suggest that gene conversion of an inactive VSG gene does not obligately activate the new VSG gene. We conclude that the gene conversion and telomeric activation mechanisms for antigenic switching are separate and independent processes. PMID- 2993881 TI - Polymorphisms within minicircle sequence classes in the kinetoplast DNA of Trypanosoma cruzi clones. AB - Four minicircle classes were analyzed using cloned minicircles as probes and single-cell cloned Trypanosoma cruzi parasites. The hybridization conditions used allowed identification of minicircle classes within kinetoplast DNA that were non homologous to each other. Two of these minicircle classes, detected with probes pTckAWP-2 and -3, were present together in several of the CA 1 and Miranda clones, in spite of the fact that either pTckAWP-2 or both minicircle classes were undetectable in other isolates and clones of the parasite. The other two minicircle classes (pTckM-84 and -88) were located in some Miranda cloned parasites which were characterized by the simple restriction endonuclease pattern of their minicircles. Both pTckM-84 and -88 minicircle classes represented 52-71% of the kinetoplast DNA in the latter group of trypanosomes. Restriction endonuclease mapping allowed the identification of polymorphic minicircles in two of the four minicircle classes analyzed (pTckAWP-2 and pTckM-88). The polymorphisms were observed in part of the molecules of one minicircle class within a single trypanosome clone, as well as when different clones or even some of those obtained from the same isolate were compared. In addition, a different proportion of pTckM-88 type of minicircle sequence class was observed in the kinetoplast DNA from two of the Miranda clones analyzed. These observations demonstrated that similar molecules may evolve independently in sequence in each parasite. The polymorphic minicircles detected may arise from sequence variations before expansion of a future homogeneous minicircle sequence class. PMID- 2993882 TI - Parasite-specific inhibition of 5'-nucleotidase from Onchocerca volvulus and Dirofilaria immitis by the amoscanate-derivative CGP 8065. AB - The presence of 5'-nucleotidase was demonstrated in Onchocerca volvulus and Dirofilaria immitis; the bulk of activity was found in the particulate fraction. The enzyme of filarial worms exhibited a broad pH-optimum between 6.4 and 8.0 and substrate specificity for nucleotides compared to glucose-6-phosphate and p nitrophenyl phosphate. The apparent Km-values for AMP were found to be 0.15 mM and 0.22 mM for the enzyme from O. volvulus and D. immitis, respectively. The activity of 5'-nucleotidase from both filarial worms was effectively inhibited by the filaricidal compound CGP 8065, a dithiocarbamate-derivative of amoscanate, whereas the 5'-nucleotidase from rat liver was not affected. The parasite specific inhibition by CGP 8065 was found to be reversible and to be competitive with respect to the substrate AMP. The inhibition constants were calculated to be 24 microM and 8 microM for the enzyme from O. volvulus and D. immitis, respectively. PMID- 2993883 TI - The primary structure of the rRNA insertions of Plasmodium lophurae. AB - The DNA sequences of the novel insertion in the 17s rRNA gene and the large insertion in the 25s rRNA gene in the cloned rDNA unit of the avian malaria parasite Plasmodium lophurae are presented, together with a partial sequence of the flanking regions, which code for the mature rRNA. The homology of the mature rRNA coding regions with the rRNA sequences of other eukaryotic organisms is extensive, indicating that the plasmodium rRNA is structurally similar to other eukaryotes. Sequence data also reveal that the region 3' to the insertion in the 17s rRNA contains a second small inserted DNA sequence, in contrast to other known small rRNA sequences. The region containing the 25s insertion shares sequence homology and some secondary structure characteristics with the terminal direct repeat of the Drosophila melanogaster transposable element copia. This is the first such sequence described in plasmodia. The direction of transcription of the cloned rDNA unit of P. lophurae has also been determined. As in other organisms, the direction of transcription is found to be 5' 17s-25s 3'. PMID- 2993884 TI - Reduced sensitivity of lymphocyte beta-adrenergic receptors in patients with endogenous depression and psychomotor agitation. AB - It has been suggested that there are altered levels of norepinephrine or other neurotransmitters at functionally important receptors in patients with depressive disorders. This hypothesis is difficult to study in the human central nervous system. However, noradrenergic function can be assessed indirectly with peripheral-blood lymphocytes used as a model of the beta-adrenergic receptor complex. We found that drug-free inpatients with endogenous depression had lower isoproterenol-stimulated cyclic AMP levels in intact lymphocytes than did healthy control subjects (3.9 +/- 0.5 vs. 7.4 +/- 1.0 pmol per 10(6) cells, P less than 0.01). The density and affinity of beta-adrenergic receptors were similar in controls and depressed subjects (beta-receptor number, 5.4 +/- 0.7 and 5.3 +/- 0.8 fmol per 10(6) cells; binding affinity, 106 +/- 7.6 vs. 99.2 +/- 11.4 pM, respectively). When the depressed patients were subdivided by psychomotor manifestations, binding characteristics were indistinguishable among the subgroups. However, a significant reduction in beta-adrenergic responsiveness was observed in patients with psychomotor agitation, as compared with controls (2.6 +/- 0.5 vs. 7.4 +/- 1.0 pmol per 10(6) cells, P less than 0.01), but not in patients with psychomotor retardation (5.8 +/- 1.1 pmol per 10(6) cells, P less than 0.05). Thus, the desensitization of beta-adrenergic receptors was correlated more closely with the severity of psychomotor agitation than with the overall severity of depression. PMID- 2993885 TI - Serum lactate dehydrogenase and bone marrow metastasis in small-cell carcinoma of the lung. PMID- 2993886 TI - Rearrangements of genes for the antigen receptor on T cells as markers of lineage and clonality in human lymphoid neoplasms. AB - The T alpha and T beta chains of the heterodimeric T-lymphocyte antigen receptor are encoded by separated DNA segments that recombine during T-cell development. We have used rearrangements of the T beta gene as a widely applicable marker of clonality in the T-cell lineage. We show that the T beta genes are used in both the T8 and T4 subpopulations of normal T cells and that Sezary leukemia, adult T cell leukemia, and the non-B-lineage acute lymphoblastic leukemias are clonal expansions of T cells. Furthermore, circulating T cells from a patient with the T8-cell-predominantly lymphocytosis associated with granulocytopenia are shown to be monoclonal. Finally, the sensitivity and specificity of this tumor-associated marker have been exploited to monitor the therapy of a patient with adult T-cell leukemia. These unique DNA rearrangements provide insights into the cellular origin, clonality, and natural history of T-cell neoplasia. PMID- 2993887 TI - Latent papillomavirus and recurring genital warts. AB - After anogenital condylomata and intraepithelial neoplasms are removed, they frequently recur. Since these lesions are related to papillomaviruses, it has been suggested that latent papillomavirus infection is responsible for recurrence. We studied 20 cases of anogenital lesions that were treated by laser therapy and analyzed biopsy specimens of margins of normal skin adjacent to the lesions for papillomavirus sequences by Southern blot hybridization. In nine cases (45 per cent), papillomavirus sequences were detected in the normal skin margin; lesions recurred in 6 of the 9 patients (67 per cent), in contrast to only 1 of 11 patients (9 per cent) whose margins were negative for the presence of papillomavirus sequences. All but one recurrence developed within 15 mm of the treatment area. Our results demonstrate that clinically and histologically latent papillomavirus exists beyond the treatment area and that its presence influences subsequent recurrences. PMID- 2993888 TI - Hemophilia A. Detection of molecular defects and of carriers by DNA analysis. AB - To understand the molecular basis of hemophilia A and to provide heterozygote detection and prenatal diagnosis by DNA analysis, we used cloned factor VIII:C DNA fragments to study 10 affected families. In four of these families, inhibitors of factor VIII:C had developed in affected persons. In one such family a deletion of approximately 80 kb within the factor VIII:C gene was identified. Carriers of the deletion were identified through detection of an abnormal DNA fragment located at the deletion end points. In another family a single nucleotide change in the coding region of the factor VIII:C gene produced a nonsense codon leading to premature termination of factor VIII:C synthesis. Carrier detection was performed in eight female members of this four-generation family. In a third family a small change in the size of a restriction endonuclease fragment correlated with the presence of the mutant gene, and in the other seven families the molecular defect has not yet been identified. In addition, we used two common polymorphic sites in the factor VIII:C gene to differentiate the normal from the defective gene in four of six obligate female carriers from families with patients in whom inhibitors did not develop. Carrier detection was possible in other members of these families. These data suggest that DNA analysis of the factor VIII:C gene provides an accurate method of carrier detection and, potentially, of prenatal diagnosis in at least 50 per cent of the pedigrees affected by hemophilia A. PMID- 2993889 TI - Hepatic encephalopathy. PMID- 2993890 TI - Receptor-mediated uptake of hypertriglyceridaemic VLDL remnants by mouse macrophages in vitro. AB - Following incubation of the VLDL isolated from hypertriglyceridaemic patients with normal human postheparin plasma VLDL remnants were obtained. 125I-VLDL remnants were taken up and catabolized by mouse macrophages in vitro. This was a receptor-dependent process. Unlabelled VLDL remnants added to the incubation medium inhibited the degradation of 125I-VLDL remnants in the cells. This could not be produced by the presence of VLDL or acetyl-LDL in the medium. It shows that mouse macrophages bind VLDL remnants by means of an other receptor than that for acetyl-LDL. The cell cholesterol content increased after incubation of the cells with VLDL remnants or acetyl-LDL, but not with native VLDL or LDL. PMID- 2993892 TI - The Harvard Group Aftercare Program: preliminary evaluation results and implementation issues. PMID- 2993891 TI - A pilot study assessing maternal marijuana use by urine assay during pregnancy. PMID- 2993893 TI - Project skills: preliminary results from a theoretically based aftercare experiment. PMID- 2993894 TI - Two British blood tests launched. PMID- 2993895 TI - US blood-bank tests established. PMID- 2993896 TI - Hormone receptor-effector complex evolution. PMID- 2993897 TI - Identity of tumour necrosis factor and the macrophage-secreted factor cachectin. AB - In mammals, several well-defined metabolic changes occur during infection, many of which are attributable to products of the reticuloendothelial system. Among these changes, a hypertriglyceridaemic state is frequently evident, resulting from defective triglyceride clearance, caused by systemic suppression of the enzyme lipoprotein lipase (LPL). We have found previously that macrophages secrete the hormone cachectin, which specifically suppresses LPL activity in cultured adipocytes (3T3-L1 cells). When originally purified from RAW 264.7 (mouse macrophage) cells, cachectin was shown to have a pI of 4.7, a subunit size of relative molecular mass (Mr) 17,000 and to form non-covalent multimers. A receptor for cachectin was identified on non-tumorigenic cultured cells and on normal mouse liver membranes. A new high-yield purification technique has enabled us to determine further details of the structure of mouse cachectin. We now report that a high degree of homology exists between the N-terminal sequence of mouse cachectin and the N-terminal sequence recently determined for human tumour necrosis factor (TNF). Purified cachectin also possesses potent TNF activity in vitro. These findings suggest that the 'cachectin' and 'TNF' activities of murine macrophage conditioned medium are attributable to a single protein, which modulates the metabolic activities of normal as well as neoplastic cells through interaction with specific high-affinity receptors. PMID- 2993898 TI - Differentiation of PC12 phaeochromocytoma cells induced by v-src oncogene. AB - PC12 rat phaeochromocytoma cells are a model system that can be used to study both neuronal differentiation and the mechanism of action of nerve growth factor (NGF). PC12 cells respond to NGF protein by shifting from a chromaffin-cell-like phenotype to a neurite-bearing sympathetic neurone-like phenotype. Here we present data on the effect of infection of PC12 cells with retroviruses carrying the src oncogene of Rous sarcoma virus. Previous studies have demonstrated that the expression of src severely affects the synthesis and accumulation of differentiated cell products in a variety of cell types. We show that in the PC12 cell system, expression of v-src appears to have an inductive effect on differentiation that resembles the action of a 'physiological' growth factor. PMID- 2993899 TI - Induction of proliferation or transformation of neuroretina cells by the mil and myc viral oncogenes. AB - The genome of the avian retrovirus MH2 contains, in addition to the v-myc oncogene shared with three other avian retroviruses (MC29, CMII and OK-10), a second cell-derived oncogene, v-mil (refs 1-3). Like the three other viruses, which contain only v-myc, MH2 induces mainly liver and kidney carcinomas in fowl and transforms fibroblasts and macrophages in vitro. However, MH2 and MC29 differ in their biological properties when assayed on cultures of chicken embryo neuroretina (NR) cells. Indeed, NR cells, which normally do not multiply in vitro, are induced to proliferate and become transformed upon infection with MH2, whereas infection with MC29 has no apparent effect on these cells. To analyse the functions of the two oncogenes of MH2, we isolated spontaneous and in vitro constructed mutants of this virus and investigated their effects on NR cell multiplication and transformation. We report here that expression of v-mil is sufficient to induce NR cell proliferation, although it does not result in cell transformation. In addition, viruses expressing only the v-myc oncogene fail to induce any detectable change in NR cells. However, cooperation of the two oncogenes is required to achieve transformation of NR cells by MH2. PMID- 2993900 TI - Nonrandom loss of chromosome 15 in Syrian hamster tumours induced by v-Ha-ras plus v-myc oncogenes. AB - Nonrandom chromosome rearrangements, observed in a variety of human and animal tumours, are associated in some cases with enhanced expression or deregulation of cellular oncogenes. Recently, it was shown that normal, diploid rodent cells are neoplastically transformed following transfection with two cooperating oncogenes, for example myc plus ras. However, the number of steps necessary to convert a normal cell into a malignant cell is unknown. If activation of two oncogenes is sufficient for tumorigenicity, tumours derived from diploid cells transformed by the transfected oncogenes may remain diploid or have only random chromosome alterations. We have performed cytogenetic analyses of tumours formed after transfection of Syrian hamster embryo cells with either v-Ha-ras plus v-myc DNAs or polyoma DNA alone. Whereas polyoma-induced, tumour-derived cells were diploid, tumours induced by v-Ha-ras plus v-myc oncogenes were monoclonal and had a nonrandom chromosome change, monosomy of chromosome 15. Thus, an additional change, loss of chromosome 15, is required for or is advantageous for tumorigenicity induced by v-Ha-ras plus v-myc oncogenes. These results suggest that the neoplastic progression of normal, diploid cells requires more than two steps under certain conditions. PMID- 2993901 TI - EGF receptor-associated DNA-nicking activity is due to a Mr-100,000 dissociable protein. AB - The receptor for epidermal growth factor (EGF) is a single-chain transmembrane polypeptide of relative molecular mass (Mr) 170,000 (170K) which has been implicated in the regulation of both normal and abnormal cell proliferation. It has an externally facing EGF-binding domain and a cytoplasmically facing tyrosine specific protein kinase site. Although the receptor has been well characterized, the mechanism by which it transmits the growth stimulatory signal from the plasma membrane to the nucleus is unclear. EGF binding to cells has been shown to enhance topoisomerase activity within the cells. Topoisomerases catalyse the interconversion of topological isomers of DNA and thus may influence replication and transcription. Mroczkowski et al. reported that purified EGF receptors of both human and murine origin can nick supercoiled double-stranded (ds) DNA in an ATP-dependent fashion, an activity related to those of topoisomerases. Another related tyrosine kinase, pp60src, has also been reported to have a similar DNA nicking activity. We have now characterized the EGF receptor-associated DNA nicking activity by sucrose gradient centrifugation. Our results, presented here, indicate that the DNA-nicking activity is not intrinsic to the EGF receptor, but is found in a distinct molecular species. PMID- 2993902 TI - Oncogenic intelligence. Another link in the chain. PMID- 2993903 TI - Amino-acid sequence of the catalytic subunit of the (Na+ + K+)ATPase deduced from a complementary DNA. AB - We have isolated and characterized a complementary DNA for the catalytic subunit of the sheep kidney sodium/potassium-dependent ATPase. The 1,016-amino-acid protein seems to have eight transmembrane domains. The apparent ouabain binding site is located at the extracellular junction of two transmembrane domains and is linked to the phosphorylation site by a 60-amino-acid conserved sequence that may be a major channel for energy transduction. PMID- 2993904 TI - Amino-acid sequence of a Ca2+ + Mg2+-dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence. AB - We have cloned and sequenced complementary DNA encoding a Ca2+-ATPase of rabbit muscle sarcoplasmic reticulum. We propose a model of the protein which has 3 cytoplasmic domains joined to a set of 10 transmembrane helices by a narrow, penta-helical stalk. In this model, ATP bound to one cytoplasmic domain would phosphorylate an aspartate in an adjoining cytoplasmic domain, inducing translocation of Ca2+ from binding sites on the stalk. PMID- 2993905 TI - Primary structure of the alpha-subunit of Torpedo californica (Na+ + K+)ATPase deduced from cDNA sequence. AB - Sodium- and potassium-dependent ATPase [(Na+ + K+)ATPase], which is responsible for the active transport of Na+ and K+, is distributed universally among animal cell membranes and consists of two types of subunits, alpha and beta (refs 1-4). The larger alpha-subunit with a relative molecular mass (Mr) of 84,000-120,000 is thought to have the catalytic role. We have now cloned and sequenced DNA complementary to the Torpedo californica electroplax messenger RNA encoding the alpha-subunit of (Na+ + K+)ATPase and have deduced the complete amino-acid sequence of the polypeptide. Some structural features of the alpha-subunit molecule related to the function of this active-transport protein are discussed. PMID- 2993906 TI - Bombesin-like peptides can function as autocrine growth factors in human small cell lung cancer. AB - The autocrine hypothesis proposes that a cell produces and secretes a hormone like substance that can interact with specific membrane receptors on its surface to induce effects such as proliferation. Thus, a cancer cell could act to stimulate its own growth. Bombesin and bombesin-like peptides (BLPs) such as gastrin-releasing peptide (GRP) cause various physiological responses in mammals, including stimulation of proliferation of 3T3 mouse fibroblasts and normal human bronchial epithelial cells in vitro and induction of gastrin cell hyperplasia and increased pancreatic DNA content in vivo in rats. Human small-cell lung cancer (SCLC) cell lines produce and secrete BLPs and can express a single class of high affinity receptors for BLPs. Exogenously added BLPs can also stimulate the clonal growth and DNA synthesis of SCLC in vitro. These findings suggest that BLPs function as autocrine growth factors for this tumour. One way to test this hypothesis is to interrupt the function of the endogenously produced BLPs. Here, we demonstrate that a monoclonal antibody to bombesin binds to the C-terminal region of BLPs, blocks the binding of the hormone to cellular receptors and inhibits the clonal growth of SCLC in vitro and the growth of SCLC xenografts in vivo. These results demonstrate that BLPs can function as autocrine growth factors for human SCLC. PMID- 2993907 TI - Unusual organization and diversity of T-cell receptor alpha-chain genes. AB - T lymphocytes recognize cell-bound antigens in the molecular context of the self major histocompatibility complex (MHC) gene products through the surface T-cell receptor(s). The minimal component of the T-cell receptor is a heterodimer composed of alpha and beta subunits, each of relative molecular mass (Mr) approximately 45,000 (refs 1-3). Recently, complementary DNA clones encoding these subunits have been isolated and characterized along with that of a third subunit of unknown function, termed gamma (refs 4-9). These studies revealed a primary structure for each subunit that was clearly similar to that of immunoglobulin and indicated a somatic rearrangement of corresponding genes that are also immunoglobulin-like. Recently, the analysis of the sequence organization of the T-cell receptor beta-chain and T-cell-specific gamma-chain gene families has been reported. We now present an initial characterization of the murine T cell receptor alpha-chain gene family, and conclude that although it is clearly related to the gene families encoding immunoglobulins, T-cell receptor beta chains and also T-cell gamma-chains, it shows unique characteristics. There is only a single constant (C) region gene segment, which is an exceptionally large distance (approximately 20-40 kilobases (kb) in the cases studied here) from joining (J) gene segments. In addition, the J cluster and the variable (V) segment number seen to be very large. Finally, in the case studied here, a complete alpha-chain gene shows no somatic mutation and can be assembled directly from V alpha, J alpha and C alpha segments without inclusion of diversity (D alpha) segments. PMID- 2993908 TI - Genomic organization of the genes encoding mouse T-cell receptor alpha-chain. AB - The vertebrate immune system uses two kinds of antigen-specific receptors, the immunoglobulin molecules of B cells and the antigen receptors of T cells. T-cell receptors are formed by a combination of two different polypeptide chains, alpha and beta (refs 1-3). Three related gene families are expressed in T cells, those encoding the T-cell receptor, alpha and beta, and a third, gamma (refs 4-6), whose function is unknown. Each of these polypeptide chains can be divided into variable (V) and constant (C) regions. The V beta regions are encoded by V beta, diversity (D beta) and joining (J beta) gene segments that rearrange in the differentiating T cell to generate V beta genes. The V gamma regions are encoded by V gamma, J gamma and, possibly, D gamma gene segments. Studies of alpha complementary DNA clones suggest that alpha-polypeptides have V alpha and C alpha regions and are encoded by V alpha and J alpha gene segments and a C alpha gene. Elsewhere in this issue we demonstrate that 18 of 19 J alpha sequences examined are distinct, indicating that the J alpha gene segment repertoire is much larger than those of the immunoglobulin (4-5) or beta (14) gene families. Here we report the germline structures of one V alpha and six J alpha mouse gene segments and demonstrate that the structures of the V alpha and J alpha gene segments and the alpha-recognition sequences for DNA rearrangement are similar to those of their immunoglobulin and beta-chain counterparts. We also show that the J alpha gene segment organization is strikingly different from that of the other immunoglobulin and rearranging T-cell gene families. Eighteen J alpha gene segments map over 60 kilobases (kb) of DNA 5' to the C alpha gene. PMID- 2993910 TI - Detection of deletions spanning the Duchenne muscular dystrophy locus using a tightly linked DNA segment. AB - The Duchenne muscular dystrophy (DMD) locus has been localized to the short arm of the human X chromosome (Xp21) by detection of structural abnormalities and by genetic linkage studies. A library highly enriched for human DNA from Xp21 was constructed using DNA isolated from a male patient who had a visible deletion and three X-linked disorders (DMD, retinitis pigmentosa and chronic granulomatous disease). Seven cloned DNA probes from this library and the probe 754 (refs 5, 8) are used in the present study to screen for deletions in the DNA isolated from 57 unrelated males with DMD. Five of these DMD males are shown to exhibit deletions for one of the cloned DNA segments and at least 38 kb of surrounding DNA. In addition, two subclones from the same region detect four restriction fragment length polymorphisms which exhibit no obligate recombination with DMD in 34 meiotic events. These new DNA segments will complement the existing Xp21 probes for use in carrier detection and prenatal diagnosis of DMD. Elucidation of the end points of the five deletions will help delineate the extent of the DMD locus and ultimately lead to an understanding of the specific sequences involved in DMD. PMID- 2993909 TI - Organization and sequences of the variable, joining and constant region genes of the human T-cell receptor alpha-chain. AB - An essential property of the immune system is its ability to generate great diversity in antibody and T-cell immune responses. The genetic and molecular mechanisms responsible for the generation of antibody diversity have been investigated during the past several years. The gene for the variable (V) region, which determines antigen specificity, is assembled when one member of each of the dispersed clusters of V gene segments, diversity (D) elements (for heavy chains only) and joining (J) segments are fused by DNA rearrangement. The cloning of the beta-chain of the T-cell antigen receptor revealed that the organization of the beta-chain locus, which is similar to that of immunoglobulin genes, is also composed of noncontiguous segments of V, D, J and constant (C) region genes. The structure of the alpha-chain seems to consist of a V and a C domain connected by a J segment. We report here that the human T-cell receptor alpha-chain gene consists of a number of noncontiguous V and J gene segments and a C region gene. The V region gene segment is interrupted by a single intron, whereas the C region contains four exons. The J segments, situated 5' of the C region gene, are dispersed over a distance of at least 35 kilobases (kb). Signal sequences, which are presumably involved in DNA recombination, are found next to the V and J gene segments. PMID- 2993912 TI - Phototransduction changes focus. PMID- 2993911 TI - Relationship between an enhancer element in the human antithrombin III gene and an immunoglobulin light-chain gene enhancer. AB - Enhancers, cis-acting transcriptional control elements have been described in both viral and cellular genes. They influence transcription in a quantitative fashion, act over relatively large distances (several kilobases, kb) and behave independently of their position and orientation. Enhancers have been described in immunoglobulin, chymotrypsin and insulin genes. They bear little homology with each other except for an 8-base pair (bp) 'consensus' core element, GTGGAAATTTG (refs 10, 11), but even this element is sometimes non-homologous. I have searched for such elements in the human antithrombin III (AT-III) gene. AT-III is an important coagulation protein which inactivates thrombin. It is produced by the liver and, to a lesser extent, by the kidney. Here, I report that the 5' flanking region of the AT-III gene encodes a segment homologous with the enhancer containing the joining-constant kappa (J kappa-C kappa) intron of immunoglobulin kappa-chain genes. This extensive homology suggests the existence of regulatory factors that recognize common DNA sequences in lymphoid tissues and in those which express AT-III. PMID- 2993913 TI - DNA topoisomerases in eukaryotes. PMID- 2993914 TI - Cyclic GMP-sensitive conductance in outer segment membrane of catfish cones. AB - A cyclic GMP-sensitive conductance has recently been observed with patch-clamp recording in excised inside-out patches of plasma membrane from frog and toad rod outer segments. This conductance has properties suggesting that it is probably the light-sensitive conductance involved in visual transduction. We now report a similar conductance in the outer segment membrane of catfish cones. Cyclic GMP showed positive cooperativity in opening this conductance, with a Hill coefficient of 1.6-3.0 and a half-saturating cGMP concentration of 35-70 microM. Cyclic AMP at 1 mM, or changing Ca concentration (in the presence of Mg), had little effect on the conductance. In physiological solutions the cGMP-induced current had a reversal potential near +10 mV; the current amplitude increased roughly exponentially with membrane potential in both depolarizing and hyperpolarizing directions. Our results suggest that cGMP is also the internal transmitter for phototransduction in cones. PMID- 2993915 TI - Cyclic GMP increases photocurrent and light sensitivity of retinal cones. AB - Like retinal rods, cone photoreceptors contain cyclic GMP and light-activated phosphodiesterase. The cGMP phosphodiesterase cascade is thought to mediate phototransduction in rods. Biochemical assays of nucleotide content in cone dominant retinas, however, have failed to demonstrate light-induced changes in cGMP. Changes in cyclic AMP following light exposure have been reported, leading to the suggestion that in cone phototransduction cAMP assumes a role analogous to that played by cGMP in rods. Cyclic GMP introduced from tight-seal pipettes into isolated cones of the larval tiger salamander, Ambystoma tigrinum, rapidly increases light-modulated membrane current more than 10-fold. In cones, as in rods, cGMP also causes an approximately 10-fold increase in photocurrent duration and a 5- to 10-fold increase in light-sensitivity. Cyclic AMP has no effect on cone photocurrents under the same conditions. Because cGMP has similar effects on photocurrent magnitude and kinetics in both rods and cones, we conclude that cGMP plays corresponding roles in transduction in both vertebrate photoreceptor classes. PMID- 2993916 TI - Mys, a family of mammalian transposable elements isolated by phylogenetic screening. AB - It has recently been demonstrated both emperically and mathematically that transposable elements may spread rapidly throughout a population once introduced even when they dramatically reduce the fitness of individuals that carry them. Such events result in pronounced differences in the phylogenetic distribution of genetic elements capable of rapid genome invasion. Using a simple and general procedure to screen the genome of the white-footed mouse Peromyscus leucopus, we have isolated a family of retrovirus-like elements which is apparently absent from the genome of the house mouse Mus domesticus. Here, we report this procedure and an analysis of the organization, phylogenetic distribution and sequence of this family of transposable elements. PMID- 2993917 TI - An exonuclease protection assay reveals heat-shock element and TATA box DNA binding proteins in crude nuclear extracts. AB - The ability to identify and purify trans-acting cellular factors that regulate eukaryotic genes is limited by the lack of a practical general assay. Current procedures using crude whole cell or nuclear extracts that restore transcriptional function in vitro or permit reconstruction of native chromatin at control sequences are effective only in select systems. I now present an exonuclease protection assay that is generally applicable for detecting sequence specific DNA-binding proteins. The assay extends earlier work on the binding to the Drosophila heat-shock gene control element of a protein factor (HAP) present in crude nuclear extracts; the binding was shown by reconstitution of specific exonuclease resistance within a nuclease-hypersensitive site in chromatin. We show here that this same exonuclease resistance can be reconstituted on free linear DNA, despite many nonspecific binding activities present in unfractionated nuclear extracts. We have further applied this assay method to fractionate the protein factor that is bound constitutively to the heat-shock gene TATA box region in native chromatin. Exonuclease protection offers a sensitive, precise and rapid assay for any sequence-specific DNA-binding protein. PMID- 2993918 TI - Crystallography attacks the cold. PMID- 2993919 TI - Molecular mechanisms of receptor desensitization using the beta-adrenergic receptor-coupled adenylate cyclase system as a model. AB - Desensitization, the tendency of biological responses to wane over time despite the continuous presence of a stimulus of constant intensity, is observed in organisms as diverse as bacteria and mammals. Recently, new insights into the molecular mechanisms underlying these phenomena have emerged from the study of the receptors coupled to the ubiquitous second messenger-generating system adenylate cyclase. These mechanisms involve sequestration or down-regulation of the receptors from the cell surface as well as functionally significant covalent modifications of the receptors and/or guanine nucleotide regulatory proteins. PMID- 2993921 TI - Simian virus 40 replication in adenovirus-transformed human cells antagonizes gene expression. AB - Simian virus 40 (SV40) replicates efficiently in monkey kidney cells. However, we have now found that SV40-based vectors transfected into most human cells replicate poorly, if at all. In contrast, strong SV40 replication is observed in human embryonic kidney (HEK) cells transformed with the adenovirus early region, but not in untransformed HEK cells. Vector replication in adenovirus-transformed cells is dependent on the presence of the SV40 origin of replication and large-T antigen. However, vigorous replication occurs at levels of large-T antigen that are undetectable by immunofluorescence. These data suggest that the adenovirus oncogenes create a replication-permissive environment to which the SV40 replicon responds. Furthermore, replication and gene expression seem to be antagonistic on our vectors. High levels of large-T antigen are observed only when vector replication is blocked by mutations in the gene for large-T antigen or the origin of replication, or by direct inhibition of DNA polymerase with aphidicolin. PMID- 2993920 TI - Structure of a human common cold virus and functional relationship to other picornaviruses. AB - We report the first atomic resolution structure of an animal virus, human rhinovirus 14. It is strikingly similar to known icosahedral plant RNA viruses. Four neutralizing immunogenic regions have been identified. These, and corresponding antigenic sequences of polio and foot-and-mouth disease viruses, reside on external protrusions. A large cleft on each icosahedral face is probably the host cell receptor binding site. PMID- 2993923 TI - Sp1 binds to promoter sequences and activates herpes simplex virus 'immediate early' gene transcription in vitro. AB - During a herpes simplex virus (HSV-1) lytic infection, three classes of viral genes are transcribed in a temporally regulated manner. The HSV-1 'immediate early' (IE) promoter sequences contain multiple copies of a hexanucleotide sequence, GGGCGG, known as a GC box, and one or more copies of an 11-base pair (bp) conserved A + T-rich element, designated TAATGARAT. The TAATGARAT elements are thought to mediate the trans-activation of IE RNA synthesis by a virion associated protein(s), and the flanking G + C-rich sequences appear both to potentiate this induction and to direct IE promoter activity in vivo. The similarity of the herpesvirus GC box repeats to those of the simian virus 40 (SV40) early promoter prompted the in vitro analysis of HSV IE transcription reported here. We show that the mammalian gene-specific transcription factor Sp1 binds to eight distinct regions of the HSV short terminal repeat and stimulates transcription 25-fold from the divergent IE-3 (ICP-4) and IE-4/5 promoters. PMID- 2993922 TI - Repression of simian virus 40 early transcription by viral DNA replication in human 293 cells. AB - The small DNA tumour virus simian virus 40 (SV40) has served as an excellent model for many studies on the mechanism and control of gene expression in eukaryotic cells. The SV40 early region produces two protein products. One product (large-T antigen) is known both to repress early viral transcription and to stimulate viral replication by binding to specific sites in the origin promoter region. The early promoter has several similarities to other RNA polymerase II promoters, for example, it possesses a TATA box, an upstream element and an enhancer. However, the SV40 early promoter differs from other known RNA polymerase II promoters in that the origin of viral DNA replication is embedded within it. Here we show that the SV40 early region is expressed at an extremely low level following its introduction ito human 293 cells, contrasting with results observed in a large number of other cells lines. We show further that the lack of expression is due to repression of transcription from the SV40 early promoter by viral DNA replication which occurs efficiently in 293 cells. PMID- 2993924 TI - Behavioral evidence for beta-adrenoceptor subsensitivity after subacute antidepressant/alpha 2-adrenoceptor antagonist treatment. AB - The behavioral consequences of beta-adrenoceptor subsensitivity were investigated by determining whether a physiological response that is mediated by beta receptors, isoproterenol-induced drinking (IID), would be reduced by subacute antidepressant/alpha 2-antagonist treatment. The coadministration of typical (e.g., imipramine) or atypical (e.g., mianserin) antidepressants with yohimbine or piperoxan twice daily for four consecutive days reduced IID. Both the time course as well as the magnitude of beta-adrenoceptor subsensitivity could be behaviorally demonstrated. In addition, the reduction in IID observed after coadministration of imipramine with yohimbine was a centrally mediated effect since it was observed after systemic (subcutaneous) and central (intraventricular) administration of isoproterenol. These results provide evidence that IID is an appropriate behavioral model to demonstrate beta adrenoceptor subsensitivity following subacute antidepressant/alpha 2-antagonist treatment. PMID- 2993925 TI - Eicosapentaenoic acid inhibits the release of 14C-prostacyclin from a perfused tissue after incorporation of 14C-prostaglandin precursors. AB - The isolated rabbit ear was labeled by perfusion either with 14C-arachidonic acid (AA) or 14C-dihomo-gamma-linolenic acid (DGLA). The influence of unlabeled eicosapentaenoic acid (EPA) on the release and metabolism of the labeled prostaglandin (PG) precursors was studied with the aid of radio-thin-layer chromatography (TLC). After incorporation of 14C-AA, the ionophore A 23187 (10 micrograms) stimulated the release of products comigrating on TLC plates with authentic PGI2 (measured as 6-keto-PGF1 alpha), PGE2, PGD2 and AA. In unlabeled ears, A 23187 (10 micrograms) stimulated the release of PGI2 and PGE2 (determined by radioimmunoassay) in similarly relative amounts as found for the labeled products. The release of PGD2 was not measured. After incorporation of 14C-DGLA, A 23187 (10 micrograms) stimulated the release of labeled products comigrating in several TLC-systems with 6-keto-PGF1 alpha (but not PGF1 alpha), PGE1, PGD1 and DGLA. After incorporation of 14C-AA, infusion of unlabeled EPA (0.1, 1 and 10 micrograms/ml) reduced the release of 14C-PGI2 but not of the other bisenoic PGs. Furthermore, in ears labeled with 14C-DGLA, EPA strongly reduced the release of a product comigrating with 6-keto-PGF1 alpha and to a lesser extent of a product comigrating with PGE1. Infusion of unlabeled EPA (1 and 10 micrograms/ml) did not reduce the release of 14C-AA or 14C-DGLA indicating that a phospholipase A2 was probably not inhibited by EPA. It is concluded that EPA inhibits PGI2 synthase rather than cyclooxygenase since after incorporation of 14C-AA, only the release of PGI2 is reduced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993926 TI - Antihypertensive mechanism of action of ketanserin and some ketanserin analogues in the spontaneously hypertensive rat. AB - Ketanserin is a new antihypertensive agent with affinity to serotonin (5-HT)2 receptors and at higher concentrations also to alpha 1-adrenoceptors. The present study was designed to evaluate the relative functional importance of the antagonism of alpha 1-adrenoceptors and 5-HT2-receptors in the antihypertensive mechanism of action of ketanserin and analogues after acute administration. In the spontaneously hypertensive rat, ketanserin and the two ketanserin analogues, R56413 and R55667 (which have relatively weaker alpha-adrenolytic properties) were studied with regard to their ability to reduce the blood pressure after acute administration in the conscious rat and their ability to shift the dose response curves for 5-HT and phenylephrine in the pithed rat. The agents tested reduced the blood pressure only in a dose range where they blocked alpha 1 adrenoceptors and there was a striking correlation between the degree of hypotension and the degree of inhibition of the phenylephrine induced pressor responses. 5-HT2-receptor blockade alone did not influence basal blood pressure. However, following pretreatment with R55667 in a low dose the blood pressure reduction to prazosin was enhanced. It is concluded that following acute administration in the rat the major portion of the antihypertensive response to ketanserin is due to an alpha 1-adrenoceptor blockade but that the 5-HT2-receptor blockade contributes. PMID- 2993927 TI - Studies on the effects of felodipine on the vascular resistance and reactivity in pentobarbital anesthetized dogs. AB - Bilateral perfused hindlimb preparations in pentobarbital anesthetized dogs were utilized in the present study to evaluate vasodilator effects of felodipine, in the presence and in the absence of sympathetic neurogenic tone. Vascular resistance was evaluated by establishing pressure-flow characteristics. A lower dose of this agent (0.01 mumol/kg i.v.) produced significant reductions in the systemic arterial blood pressure and vascular resistance only in the innervated but not in the denervated limb. This effect was shown to be due to interference with noradrenaline induced vasoconstriction. However, a tenfold higher dose of felodipine (0.1 mumol/kg i.v.) was indeed effective in attenuating resistance in the denervated vascular bed; this ability of the higher dose (0.1 mumol/kg) to inhibit vascular resistance may be due to an additional intracellular action of this drug affecting myogenic tone and/or due to inhibition of the vascular effects of the endogenous renin-angiotensin system. In the present experimental model, felodipine, a dihydropyridine calcium antagonist, was equally effective in inhibiting either alpha 1- or alpha 2-mediated vasoconstriction. These data suggest that the ability of felodipine to interfere with sympathetic vasoconstrictor mechanisms primarily contributes to its hypotensive effects in the barbiturate-anesthetized dogs. PMID- 2993929 TI - Learning experiences from UNC. Renal autoregulation, drugs, and acute renal failure. PMID- 2993930 TI - [Wart virus and cancer]. PMID- 2993931 TI - Lymphoid malignancies in Nebraska: a hypothesis. PMID- 2993932 TI - Adult T-cell leukaemia lymphoma (ATLL) associated with human T-cell leukaemia virus (HTLV). Case report and immunological studies with the neoplastic T-cells of a Caribbean patient. PMID- 2993933 TI - Hepatic failure due to liver metastases in breast cancer. PMID- 2993934 TI - Effects of purified hog pancreatic kallikrein on the kinin-prostaglandin system and renin-angiotensin-aldosterone system. AB - In the present study, hog pancreatic kallikrein (HPK, 2,000 KU/kg body weight) was intramuscularly injected into male rabbits and several plasma hormones [kinin, prostaglandin E (PGE), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), thromboxane B2 (TXB2), plasma renin activity (PRA), aldosterone, and ACTH] were measured before and after the HPK administration, in order to clarify the role of glandular kallikrein in the blood. The plasma kinin concentrations were significantly (p less than 0.001) increased from 1 +/- 1 (mean +/- SE), the baseline level, to 230 +/- 22, 288 +/- 36 and 130 +/- 9 pg/ml at 30, 60, and 120 min, respectively, after HPK administration. The plasma level of PGE were slightly increased after HPK administration, but the change was not significant compared to the mean baseline level. The plasma levels of 6-keto-PGF1 alpha were significantly increased from 229 +/- 38, the baseline level, to 594 +/- 131 (p less than 0.05) at 30 min after the administration of HPK and tended to increase (378 +/- 67) at 60 min after the HPK administration, but fell to 278 +/- 37 pg/ml at 120 min after the administration of HPK. On the other hand, the changes in plasma TXB2, aldosterone, ACTH, and PRA before and after HPK administration were not significant. The present results showed that exogenous intramuscular administration of HPK increased the plasma levels of kinin and prostacyclin, but caused no elevation in the plasma levels of other hormones including PRA. It is concluded, therefore, that in this acute experiment, there was a close relationship between the kallikrein-kinin system and prostaglandins. PMID- 2993928 TI - Effects of the irreversible alpha-adrenoceptor antagonists phenoxybenzamine and benextramine on the effectiveness of nifedipine in inhibiting alpha 1- and alpha 2-adrenoceptor mediated vasoconstriction in pithed rats. AB - In pithed normotensive rats, i.v. injection of the selective alpha 1-adrenoceptor agonist cirazoline produced vasoconstriction which was largely resistant to inhibition by nifedipine. On the other hand, the pressor effects of the selective alpha 1-adrenoceptor agonists St 587 and Sgd 101/75 were much more effectively blocked by nifedipine, although not as effectively as the pressor effects to the selective alpha 2-adrenoceptor agonist B-HT 920. The sensitivity to inhibition of vasoconstriction in pithed rats to the different agonists increased in the order cirazoline much less than St 587 less than Sgd 101/75 less than B-HT 920. Phenoxybenzamine (3-300 micrograms/kg, i.v., -60 min) irreversibly antagonized the vasoconstriction to cirazoline, St 587, Sgd 101/75 and B-HT 920. After treatment of the rats with phenoxybenzamine the potency and efficacy of nifedipine in antagonizing vasoconstriction to alpha 1-, but not to alpha 2 adrenoceptor activation was dose-dependently enhanced. The potency of nifedipine to inhibit alpha 1-adrenoceptor-mediated vasoconstriction by cirazoline, St 587 and Sgd 101/75 was increased maximally to the level of efficacy at which nifedipine antagonized B-HT 920-induced vasoconstriction. The dose of phenoxybenzamine required to maximally increase the potency and efficacy of nifedipine to antagonize vasoconstriction of the alpha 1-adrenoceptor agonists was inversely related to the level of sensitivity to blockade by nifedipine of the vasoconstriction they produced. In contrast, pretreatment of rats with the irreversible antagonist, benextramine (10 mg/kg, i.v., -100 to -60 min) did not increase the potency or efficacy of nifedipine to antagonize vasoconstriction to cirazoline, St 587, Sgd 101/75 or B-HT 920, despite irreversible blockade of alpha 1- and alpha 2-adrenoceptors. These data suggest that phenoxybenzamine, but not benextramine, selectively inhibits the alpha 1-adrenoceptor mediated vasoconstrictor mechanism that is independent of influx of extracellular calcium. Moreover, the results show that the existence of receptor reserve or the number of alpha 1-adrenoceptors activated does not determine the relative contribution of calcium influx-independent mechanisms in alpha 1-adrenoceptor-mediated vasoconstriction. PMID- 2993935 TI - Antinociceptive effect of diisopropylphosphofluoridate: development of tolerance and lack of cross-tolerance to morphine. AB - The irreversible cholinesterase inhibitor diisopropylphosphofluoridate (DFP) causes a naloxone-sensitive antinociceptive effect in laboratory animals. Chronic treatment of male mice with DFP (2 mg/kg/day for fourteen days) rendered the animals tolerant to its antinociceptive effect. Animals tolerant to DFP were cross-tolerant to the antinociception induced by the cholinergic agonists oxotremorine and nicotine, but no cross-tolerance with morphine was observed. Similarly, mice made tolerant to morphine were not cross-tolerant to DFP, nor were they cross-tolerant to oxotremorine and nicotine. Binding of muscarinic and nicotinic cholinergic ligands was significantly decreased in the brain of DFP tolerant mice, due to a reduction in receptor density. No change was observed in the binding of [3H]-dihydromorphine to opiate receptors. None of these three binding sites was altered in mice tolerant to morphine. Although there is evidence of an involvement of endogenous opioids in the antinociceptive action of DFP, the lack of cross-tolerance between DFP and morphine suggests the existence of a more complex interaction between DFP and the cholinergic and opiate systems. PMID- 2993936 TI - Proteolytic activity in brains of rabbits treated with aluminum. AB - Aluminum injection in rabbits leads to neurofibrillary changes which are at light microscopic level similar to those found in Alzheimer's disease. We used this animal model to see whether changes in proteolytic activity occur that may affect protein degradation in the altered neurofibrillary structure. Rabbits were injected via the cisterna magna with aluminum chloride, and after ten days tissue was excised from the spinal cord, hippocampus, occipital lobe, and cerebellum. Sections from the hippocampus and spinal cord were examined for neurofibrillary changes; enzyme activity was measured in all four areas. The enzymes studied were cathepsins A, B, and D, and the angiotensin-converting enzyme. No significant differences could be established in enzymatic activity in aluminum-injected animals compared to controls. However, a significant decrease in Triton-soluble proteins was observed in the treated animals, which correlated with changes in neurofibrillary structure. This decrease was most noticeable in the spinal cord (from 16.6 to 12.5 mg/g). PMID- 2993937 TI - Synthesis of thiamine triphosphate in rat brain in vivo. AB - Thiamine metabolism in vivo was studied by intracerebroventricular injection of labeled thiamine in rat brain. Labeled thiamine was found to be rapidly converted to the phosphorylated thiamine esters. The distribution of the radioactive thiamine compounds was reached to steady state at 3 hr after injection: thiamine, thiamine monophosphate, thiamine pyrophosphate, and thiamine triphosphate were 8 12%, 12-14%, 72-74%, and 2-3%, respectively, in cerebral cortex. The presence of labeled thiamine triphosphate in the brain was further confirmed by the treatment with thiamine triphosphatase which had an absolute substrate specificity for thiamine triphosphate. These results suggest that thiamine triphosphate is synthesized in vivo in rat brain. PMID- 2993939 TI - Localization and heterogeneity of central benzodiazepine receptors. PMID- 2993938 TI - Further characterization of [3H] flunitrazepam binding sites on cultured mouse astroglia. AB - Astroglial cells in primary cultures bind [3H]flunitrazepam with a high affinity on a single type of site and on a number of binding sites which increased during astroglial growth and differentiation. These binding sites show a particular pharmacological spectrum characterized by an inhibition of high affinity by RO-5 4864 (4'-chlorodiazepam), an anticonvulsant of the benzodiazepine family and by an inhibition of binding of lower affinities by diazepam clonazepam and clobazam. RO-5-4864 and clonazepam compete for the same binding site in astroglia. The heat stability and the hormonal modulation by thyroxine are similar for astroglia and neuronal-cells. Benzodiazepines modulate the astroglial 5-HT receptor. Such an effect could be a possible physiological response to benzodiazepines for astroglial cells in primary cultures. PMID- 2993940 TI - Changes of 5'-nucleotidase activity in cultured glioma cells. AB - Changes of 5'-nucleotidase (5'-N) activity during subcultivation of C6 clonal rat glioma cells were studied histochemically, cytochemically and biochemically. Two different cell populations were seeded with high or low cell density respectively. 5'-N activity in cultured cells increased continuously in both populations. The enzyme activity was always higher in the cells growing in high density than in those growing in low density, and this difference became statistically significant after 6 days of subcultivation. It seemed likely that the increase in 5'-N activity depended on cell density, not on the frequency of mitosis. Cytochemically, enzyme activity was detected on the outer surface of the plasma membrane of cell bodies or cell processes. Strong activity was found on the processes and cell membranes where cells touched each other. In the regions where cells had intimate cell-to-cell contact, marked elevation of the enzyme activity was seen. Increased 5'-N activity, which appeared to depend on cell density, was considered to be related with this cell-to-cell contact. PMID- 2993941 TI - Transient uncrossed corticospinal fibres in the newborn rat. AB - A small number of uncrossed corticospinal fibres were demonstrated in a 1-day-old rat by injection of horseradish peroxidase into one hemisphere of the cerebral cortex. These fibres were not demonstrable after the age of 7 days. PMID- 2993942 TI - Acrylamide-induced ascending degeneration of ligated peripheral nerve: effect of ligature location. AB - Acrylamide was administered to rats for 10 days (50 mg/kg/day) after ligation of either proximal (sciatic) or distal (posterior tibial) nerves. Morphologic evaluation of the nerves revealed severe degeneration 0.5 cm proximal to the ligature, with progressively less pronounced change at successively higher levels above the ligature, when compared with ligated non-treated controls. Although severe degeneration of the nerves was observed in both proximal- and distal ligated groups of animals, the distal-ligated group showed slightly more change (at 0.5 cm above the ligature), suggesting a minor effect of distance of the site of nerve injury from the nerve cell body. PMID- 2993943 TI - Ipsilateral sensory symptoms caused by a temporal lobe glioma. AB - Sensory symptoms can occasionally occur ipsilateral to lesions of the insula. We report a patient with left-sided sensory symptoms preceding the diagnosis of a left temporal lobe glioma. The anatomical basis for this clinical phenomenon is reviewed. This patient illustrates the complex neuroanatomical pathways and the clinical correlations from lesions in this region. PMID- 2993944 TI - Neuropeptides in human memory and learning processes. AB - The neuropeptides vasopressin, adrenocorticotropin (ACTH), and beta-endorphin seem to have important effects on memory and learning. Animal studies attempting to demonstrate these effects are difficult to interpret because of the complexity of behavior that is described as "learning" and the impossibility of assessing verbal learning in animals. This article therefore reviews some of the animal literature on neuropeptides and learning, but focuses primarily upon studies in humans, both in normal volunteers and in patients with neurological disorders. Vasopressin enhances learning under some conditions. Intranasal administration has been associated with improvement on psychometric tests in patients with mild Alzheimer's disease and Korsakoff's psychosis, although these findings are not uniform. It improves performance on memory tests in normal volunteers, but does not seem to improve the memory deficit after head trauma. Cerebrospinal fluid levels are low in patients with Alzheimer's disease. ACTH and melanocyte stimulating hormone (MSH) are two peptides the primary behavioral effect of which seems to be on attention or goal-motivated behavior rather than on memory processes themselves. Visual discrimination and the ability to continue repetitive tasks are enhanced; in mentally retarded subjects, the administration of ACTH or MSH improves performance on a variety of neuropsychological tests. It does not, however, improve cognitive function in the elderly. Endogenous opioids including beta-endorphin and met-enkephalin seem to have primarily an amnesic effect in animal studies. Their role in human learning is still uncertain, although naloxone, which antagonizes their effects, has been associated with improved cognitive performance in patients with Alzheimer's disease. These data underscore the complexity of the processes associated with human memory and the rudimentary state of our present knowledge. Whatever the mechanisms, however, vasopressin, ACTH, and endogenous opioids seem to have important effects upon memory. PMID- 2993945 TI - Brain noradrenergic activity in wakefulness and paradoxical sleep: the effect of clonidine. AB - Several experiments in which the effects of clonidine (CLN) have been studied in the rat and in man are summarized. CLN inhibits paradoxical sleep (PS) both in the rat and in man. This inhibition is followed by a rebound, but with a different time course in the two species. Wakefulness (W) is enhanced after some critical doses of CLN, and decreased after higher doses. These effects of CLN suggest that alpha 2-adrenoceptors are involved in the regulation of W and PS. PMID- 2993946 TI - Early visual information processing in depressive patients treated with ORG 2766 (an ACTH 4-9 analogue). AB - Twenty-one depressive patients (7 males, 14 females, mean age 61.6, range 50-72) participated in an inter- and intraindividual double-blind comparative study between the ACTH 4-9 analogue (ORG 2766), 80 mg daily during 14-28 days, and placebo. The temporal thresholds for detection and recognition of rapidly presented single letters were assessed by a tachistoscopic technique. The results indicate that the peptide has a reducing effect on recognition threshold, and appears to improve automatic, preattentive levels of information processing possibly involved in the transfer of information from the peripheral icon to the cortical centre, i.e. from iconic storage to short-term memory. PMID- 2993947 TI - Intracerebroventricular administration of cholecystokinin inhibits the activity of the dopaminergic and serotoninergic systems of the brain. AB - The behavioral and biochemical effects of intracerebroventricular administration of cholecystokinin were investigated in experiments on male Wistar rats. Cholecystokinin induced specific dose-dependent changes in the behavior of the animals. At low doses the inhibiting influence on behavior predominated; at high doses stereo-typed behavior, shaking of the head and increased reactivity to pain stimuli were observed. Cholecystokinin appreciably inhibited the circulation of serotonin and dopamine in the brain structures in comparison with physiological saline solution. Administration of cholecystokinin against a background of phenamine and 5-hydroxy-tryptophan briefly entirely inhibited the behavioral effects induced by these substances. On the basis of the data obtained it can be assumed that cholecystokinin is an endogenous modulator of the activity of the monoaminergic systems of the brain. PMID- 2993948 TI - Electrophysiologic quantification of distal polyneuropathy in diabetes. AB - Electrophysiologic studies of the lower limb (reflex, motor, and sensory) were done in 50 diabetics, 30 of whom were age- and sex-matched with controls. The results were analyzed by a 0 to 9 rating--electrophysiologic polyneuropathy (EPN) score. The median EPN scores were 0.5 in controls and 3.5 in diabetics (p less than 0.001). Diabetics with mild or no distal polyneuropathy (DPN) had a median EPN score of 2.4 compared with 4.7 in those with overt DPN (p less than 0.0001). The EPN score correlated positively with known duration of diabetes and showed differences with age and therapy. It provides a single electrophysiologic parameter for evaluating DPN in diabetics. PMID- 2993949 TI - Sensory neuropathy with low-dose pyridoxine. AB - We describe 16 patients with neuropathy associated with pyridoxine abuse. The clinical picture of a pure sensory central-peripheral distal axonopathy was consistent. Pyridoxine dose was 0.2 to 5 g/d, and duration of consumption before symptoms was inversely proportional to the daily intake. In all patients with adequate follow-up, improvement followed discontinuation of pyridoxine. The ready availability of up to 1-gram tablets makes it likely that this neuropathy will continue to be seen. PMID- 2993950 TI - Guillain-Barre syndrome after epidural anesthesia: direct nerve root damage may trigger disease. AB - Guillain-Barre syndrome (GBS) appeared in four patients 1 to 2 weeks after epidural anesthesia. In all patients, clinical diagnosis was confirmed by CSF findings and nerve conduction velocity studies. Although epidural anesthesia has not been listed as an antecedent event in GBS, evidence for the relationship has been previously reported. Interaction between the anesthetic agents and peripheral nervous system myelin or local trauma to roots may initiate a cascade of immunologic events that result in the demyelinating neuropathy. PMID- 2993951 TI - Inflammatory neuropathy in homosexual men with lymphadenopathy. AB - Twelve homosexual men had peripheral neuropathy with fever, night sweats, and lymphadenopathy. Sensory symptoms predominated, but there was also weakness and cranial nerve dysfunction. Manifestations were multifocal in nine and distal and symmetric in three. CSF was abnormal in all eight patients examined. Sural nerve in five patients showed axonal degeneration, accompanied in two by segmental demyelination. Four patients had epineurial and endoneurial perivascular chronic inflammatory cells without evidence of vasculitis. Neuropathy remitted spontaneously in six patients. Four patients received steroids without clinical response, although one later responded to plasmapheresis-lymphocytapheresis. Four patients later progressed to AIDS. PMID- 2993952 TI - Electrophysiologic evaluation of lumbosacral radiculopathies: electromyography, late responses, and somatosensory evoked potentials. AB - We compared the diagnostic utility of EMG, F wave and H-reflex studies, and peroneal and dermatomal SEPs in evaluating 28 patients with clinically unequivocal L-5 or S-1 compressive root lesions. The single most useful electrophysiologic technique was EMG, which often provided evidence of denervation in a myotomal pattern when other electrophysiologic findings were normal. We found abnormal late responses in 14 patients, but always in association with EMG abnormalities. Peroneal-derived SEPs were always normal. Dermatomal SEPs confirmed the diagnosis in seven patients, including two in whom other electrophysiologic studies were normal. PMID- 2993953 TI - [Functional relations between neurohormonal and immunological systems]. PMID- 2993955 TI - Diagnosis, treatment and postoperative results of Cushing's disease. AB - One hundred patients with Cushing's disease were operated on in Nagoya University Hospital between January 1977 and January 1984. Pituitary adenomas were found and resected in 93 cases and complete clinical remissions were observed in 89 treated by operation alone and in two additional cases by operation followed with radiotherapy. The clinical features, pre-operative endocrine data, radiological findings including high resolution CT with coronal and sagittal reconstructions, data of selective venous sampling, operative findings, tumour pathology, postoperative clinical course, postoperative endocrine results and follow-up findings are presented and discussed. PMID- 2993954 TI - Corticotropin- and growth hormone-releasing factor (CRF and GRF) in the diagnosis of hypothalamo-pituitary diseases. AB - The recently discovered releasing factors for ACTH and growth hormone, CRF and GRF, have stimulated our understanding of the hypothalamo-pituitary physiology and pathophysiology. Whereas CRF, a 41 amino-acid residue peptide, seems to be a useful additional tool in the differential diagnosis of Cushing's syndrome, GRF, a 44 amino-acid residue peptide, seems to be of little value in the diagnosis of active acromegaly. However, both releasing factors are useful in the follow-up of these two hypersecretory diseases after neurosurgical treatment. In addition, the measurement of CRF and GRF activity by radioimmunoassay in the peripheral circulation will be of great importance in making the diagnosis of ectopic CRF- and GRF-secretion, syndromes which have been appreciated only in recent years. Furthermore, CRF and GRF-stimulation tests are useful in elucidating the cause of anterior pituitary failure and GRF may be of benefit for the treatment of pituitary dwarfism. However, it has to be stressed that the results of the new CRF and GRF tests can only be interpreted together with results of other established pituitary function tests and the neuroradiological evaluation of the hypothalamo-pituitary area. PMID- 2993957 TI - [Glucagonoma without the glucagonoma syndrome]. PMID- 2993956 TI - Trans-sphenoidal microsurgical treatment of Nelson's syndrome. AB - Clinical features and the operative results in the author's personal series of ten cases with Nelson's syndrome are presented. There were nine females and one male patient with ages ranging from 20 to 44 years (mean 36 y). All patients presented with typical hyperpigmentation of the skin and had abnormally raised plasma ACTH levels. X-ray polytomography detected sellar abnormalities in 80% of the cases. Computerized tomography was positive for pituitary tumour in three patients. Trans-sphenoidal microsurgical removal of Nelson's ACTH adenomas were successful in seven of ten patients. The operative technique of radical-selective resection to achieve endocrinological cure is described. Pertinent literature is reviewed and the incidence of Nelson's syndrome, an issue of controversies regarding the pathogenesis, and the management of Nelson's adenomas are discussed. PMID- 2993958 TI - [Therapy of fissure in ano]. PMID- 2993959 TI - [Effects of subcutaneous administration of pentagastrin and duodenal alkalinization on plasma group I pepsinogen (PGI-p) in man]. PMID- 2993960 TI - [Dietetic and environmental factors in constipation in urban and rural populations of Western Sicily]. PMID- 2993962 TI - [Changes in the circadian rhythm of urinary cyclic adenosine monophosphate during the alcoholic withdrawal syndrome and related neurobiological correlations]. AB - Urinary cyclic adenosine monophosphate (AMP-c) was measured morning and evening in 35 patients with alcohol withdrawal syndrome (AWS). 65% of the patients revealed a higher night time than day time concentration of AMP-c in the urine, reflecting increased sympathetic adrenergic activity. The circadian rhythm was lost in 88.55% of the 35 patients. The pathogenic factors and mechanisms involved in AWS are discussed and the contribution of sympathetic adrenergic hyperactivity to the onset of the withdrawal syndrome with its concomitant depression of the cholinergic and GABAergic systems is emphasised. Finally it is suggested that insomnia and the loss of REM sleep may also contribute to the onset of the condition. PMID- 2993961 TI - [Maternal drug dependence and infections. Perinatal mortality]. AB - Some toxic and infectious factors affecting pregnant drug addicts are analysed. Harmful effects on the foetus and the newborn are assessed. The problem of perinatal mortality as a result of maternal drug addiction is examined. A review of the literature reveals the mortality rate to be high. PMID- 2993963 TI - Binding of [3H]cyclohexyladenosine to adenosine recognition sites in guinea pig ileal membranes: comparison with binding in brain membranes. AB - The stable adenosine agonist [3H]cyclohexyladenosine binds to membranes prepared from guinea pig myenteric plexus with high affinity (Kd = 1.8 nM) in a saturable, reversible manner. Binding pharmacology was consistent with the labelling of an A1 type adenosine receptor with the R-isomer of N6-phenylisopropyladenosine (PIA) being 11 times more potent than the S-diastereomer. Binding was similar to that observed in brain tissue being displaced by a number of xanthine adenosine antagonists. In terms of density of binding sites, however, there were approximately 9 times fewer higher affinity binding sites in ileal membranes than in the central nervous system. PMID- 2993964 TI - Calcitonin gene-related peptide is present in mammalian cerebrovascular nerve fibres and dilates pial and peripheral arteries. AB - Calcitonin gene-related peptide (CGRP) is a novel 37-amino acid peptide occurring in neurones within sensory ganglia, in brain stem, as well as in the walls of blood vessels of peripheral organs. Pial arteries of cat showed a well-developed supply of CGRP-positive nerve fibres. The peptide was found to be a potent dilator of both pial and peripheral vessels of rabbit and cat, and of pial vessels from man. The dilatory effect was independent of the vascular endothelium and was not mediated through adrenergic, cholinergic or histaminergic smooth muscle receptors. The neurogenic vasoconstriction induced by electrical field stimulation was temporarily inhibited by CGRP, as studied in central ear arteries from rabbits. The results suggest that CGRP is a transmitter or modulator playing a role in the regulation of vascular tone. PMID- 2993965 TI - Spontaneous inhibitory postsynaptic potentials in guinea pig neocortex and olfactory cortex neurones. AB - The membrane potential of olfactory cortex and neocortex neurones in vitro was recorded using conventional microelectrode techniques. During recordings with KCl or CsCl-filled microelectrodes, spontaneous, subthreshold, transient membrane depolarizations were observed. These were abolished by the GABAA-receptor antagonist, bicuculline methiodide, and were prolonged by the barbiturate pentobarbitone. In most cells they were abolished by tetrodotoxin. It is concluded that these spontaneous depolarizations are inhibitory postsynaptic potentials arising from spontaneous activity in inhibitory interneurones. PMID- 2993966 TI - Frequency-to-voltage conversion in the pyramidal tract neuron: an important role of the inhibitory postsynaptic potential. AB - Frequency-coded impulses are known to be converted into postsynaptic potentials (PSPs) at the synapse of a target neuron. This can be termed frequency-voltage (F V) conversion. Studies on this problem in pyramidal tract neurons (PTNs) showed that not only the amplitude but also the duration of depolarizing PSPs was determined as a function of the input impulse frequency. Two opposite patterns of F-V conversion were observed following activation of two input systems to PTNs. Inhibitory postsynaptic potentials were found to play an important role in the regulation of the duration of PSPs by curtailing excitatory post-synaptic potentials. PMID- 2993967 TI - Electrophysiological properties of single units in dopamine-rich mesencephalic transplants in rat brain. AB - Electrophysiological recordings were made from grafts of embryonic ventral mesencephalon transplanted into the neocortex, overlying the host neostriatum of rats in which the intrinsic striatal dopamine (DA) innervation had been removed by a unilateral 6-hydroxydopamine (6-OHDA) lesion. Whereas all rats had surviving grafts, half of the rats showed recovery from amphetamine-induced rotational asymmetry induced by the 6-OHDA lesions ('compensation'), indicating a functional dopaminergic reinnervation of the host striatum by the graft. The electrophysiological recordings revealed: neurons within all grafts which could be antidromically activated from host striatum, and which had faster conduction velocities and narrower action potentials than is characteristic of DA neurons in the intact substantia nigra, neurons only within the grafts of compensated rats which had properties characteristic of normal nigral DA neurons, and neurons within the grafts which responded to stimulation of the frontal cortex and lower brainstem of the host. The data support the view that the grafts can establish physiologically functional afferent and efferent connections with the host brain. PMID- 2993968 TI - Changes in brainstem and spinal adrenoceptor binding with ageing in spontaneously hypertensive and Wistar-Kyoto rats. AB - Adrenoceptor binding in membranes prepared from the brainstem and thoracic spinal cord of male spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats aged 6 and 36-40 weeks has been compared. Binding of [3H]prazosin (alpha 1-receptors), [3H]rauwolscine (alpha 2-receptors) and [3H]dihydroalprenolol (DHA: beta receptors) fell with age in both regions in both strains. Strain, independent of age, was also a significant determinant of binding for all three ligands in brainstem and for [3H]DHA in spinal cord. The changes in receptor binding may reflect differences in noradrenergic activity with age and between SHR and WKY rats. PMID- 2993969 TI - Antagonistic effect of delta-aminovaleric acid on bicuculline-insensitive gamma aminobutyric acid B (GABA B) sites in the rat's brain. AB - The effect of delta-aminovaleric acid (delta-AV) on bicuculline-insensitive gamma aminobutyric acid B (GABA B) sites in the central nervous system (CNS) was investigated by binding studies and experiments on slices in vitro. delta-AV inhibited [3H]GABA (10 nM) binding to GABA B sites in a rat brain membrane preparation with an IC50 value of 10(-4) M. It also inhibited [3H]baclofen (20 nM) binding with an IC50 value of 10(-4) M. In preparations of hippocampal slices, (-)-baclofen (5 microM) reduced the population spikes evoked by stimulating the Schaffer collaterals in CA1 pyramidal cells in the presence of 100 microM bicuculline. delta-AV (1 mM) antagonized this inhibitory action of baclofen. Since baclofen is an agonist of GABAB sites, our results indicate that delta-AV has an antagonistic effect on GABAB sites in the CNS. PMID- 2993971 TI - The gamma-aminobutyric acid-mimetic action of omega-aminocaprylic acid in rat cortex. AB - The 8-carbon chain analogue of gamma-aminobutyric acid (GABA), omega aminocaprylic acid (omega-AC), was administered to single cortical neurones of rat by microiontophoresis and its effect on cell firing compared to the effect elicited by GABA. The spontaneous and glutamate-elicited firing rates of all cells examined were decreased by omega-AC, the molecule having an apparent potency approximately one-fifth that of GABA. Inhibitions induced by omega-AC were antagonized by bicuculline and enhanced by flurazepam. These data favour the view that extending chain length per se of the GABA molecule decreases the inhibitory potency of postsynaptic GABA receptor agonists. The results do not support the view that such structural changes alter the mode of action of GABA analogues, from inhibition to excitation, as has been suggested previously. PMID- 2993970 TI - Benzodiazepine receptors are not involved in the neurotoxicity and anti electroshock activity of phenytoin in mice. AB - The present study was undertaken to investigate whether benzodiazepine receptors are involved in either the anticonvulsant activity of phenytoin against supramaximal seizures or maximal threshold seizures induced by electroshock or the neurotoxicity of phenytoin. Ro 15-1788, a benzodiazepine receptor antagonist, reversed the anticonvulsant activity of clonazepam against both supramaximal and maximal threshold seizures induced by electroshock but not that of phenytoin. Moreover, Ro 15-1788 did not decrease the neurotoxicity of phenytoin. These results suggest that benzodiazepine receptors are not involved in either the anticonvulsant action of phenytoin against seizures induced by electroshock or the neurotoxicity of phenytoin. PMID- 2993972 TI - Localization of ganglioside GM1 on myelin, in dissociated cells from rat embryonic cerebral hemispheres, using biotinylated choleragen and avidin peroxidase. AB - The distribution of ganglioside GM1 in primary cultured cells from rat cerebral hemispheres was studied. These dissociated cell cultures are capable of forming myelin. 2',3'-Cyclic nucleotide 3'-phosphodiesterase (CNPase) activity, which is considered to be a good marker for myelin, increased with the onset of myelin formation. Ganglioside GM1 was localized on the myelin-related structures using a biotinylated choleragen and avidin peroxidase technique. The distribution of ganglioside GM1 resembled that of immunostained CNPase. PMID- 2993973 TI - Influence of sexual hormones on solid tumor growth and ATP C+ cell replication. AB - Solid tumors obtained by implanting ATP C+ cells subcutaneously into the back of BALB/cf/Had/Se substrain mice developed more rapidly in males than females. Experiments conducted on gonadectomized animals demonstrated that female gonads inhibit the development of this tumor. Studies on ATP C+ cells cultured in vitro with various doses of male and female hormones showed that mainly progesterone, but to a lesser extent also estradiol, significantly inhibit cell proliferation with respect to testosterone. The treatment of normal and gonadectomized animals with testosterone and progesterone confirms the results obtained in vitro. Analyses carried out on ATP C+ cells do not reveal presence of estradiol and progesterone receptors. PMID- 2993974 TI - Retinal vascular tortuosity and recurrent retinal haemorrhages in a patient with non-chromaffin paraganglioma. AB - The present report describes the history of a 66-year-old man suffering from recurrent retinal haemorrhages combined with retinal vascular tortuosity. He also developed a non-chromaffin paraganglioma in the neck, and the coincidence of these two rare diseases is discussed. PMID- 2993976 TI - Glomus tumor of the cheek: an immunohistochemical demonstration of actin and myosin. AB - A rare case of a buccal glomus tumor was examined by light and electron microscopy. Histologically, the tumor consisted of various vascular spaces surrounded by masses of epithelioid glomus cells, coexisting with a hemangiomatous proliferation of the vessels. Immunohistochemical stainings revealed that the tumor cells were positive for smooth-muscle actin and myosin, with various intensities of the stainings. The electron microscope invariably showed microfilaments in the tumor cells. The results suggest that the glomus tumor cells have characteristics of the smooth-muscle cell and the tumor has a hamartomatous nature. PMID- 2993975 TI - Prolactin binding in minor salivary gland tumors. AB - An immunohistochemical study of 15 minor salivary gland tumors was initiated to determine if prolactin binding occurred in these tissues. Eight benign mixed tumors (BMT) and 7 adenoid cystic carcinomas (ACC) were selected at random from the surgical biopsy service of the MCV/VCU School of Dentistry, Department of Oral Pathology. The specimens were cut and mounted on slides along with sections of rat pituitary and rat ventral prostate which served as methodologic controls. Experimental specimens were incubated for 24 hours with varying concentrations of highly purified (iodination grade) rat prolactin; controls were incubated with vehicle. Following incubation the specimens were stained according to the Sternberger peroxidase-antiperoxidase method. Results showed dose-dependent staining for prolactin binding sites in 7 of 8 BMTs and 5 of 7 ACCs. The staining was wider in distribution than we observed in normal human minor salivary gland tissue. Binding was confined primarily to cells of duct origin in both types of tumor. In individual cells, staining was observed in diffuse cytoplasmic and perinuclear locations as well as in nuclei and apical regions. We conclude that two minor salivary gland neoplasms (BMT and ACC) exhibit prolactin binding at different cellular locations and in a more widespread pattern than was observed in normal minor salivary gland. PMID- 2993977 TI - Papillomaviruses: their possible role in oral disease. AB - Papillomaviruses are ubiquitous DNA viruses that are epitheliotropic and produce a range of epithelial neoplasms, both benign and malignant, in animals and man. Human papillomaviruses are associated with a variety of rare and uncommon oral lesions, and there has been increasing suspicion that they may be implicated also in some premalignant and malignant oral lesions. PMID- 2993978 TI - Ridge form in preprosthetic surgery. AB - In preprosthetic surgery there is a need for improved methods of diagnosis that are based on the recognition of the anatomy of the deficient residual ridge. These methods enable the surgeon to direct his attention to obtaining the ideal ridge form for denture construction. To this end, characteristics and patterns of resorption as well as a cephalometric method of assessment are presented. A problem-oriented classification of deficient residual ridges will also permit the clinician to select the most effective method of treatment. PMID- 2993979 TI - [Should healthy breast-fed infants, born at term, be given vitamin D?]. PMID- 2993980 TI - [IIB-type Willebrand disease (a case of increased ristomycin-induced aggregation)]. PMID- 2993981 TI - [A successfully treated case of spironolactone-induced hyperkalemic quadriplegia]. PMID- 2993982 TI - [Vasoactive intestinal polypeptide (VIP)--possible importance in diseases of childhood]. AB - Vasoactive intestinal peptide (VIP), first isolated from the gut, was originally considered a candidate gastrointestinal hormone. Since about 1975, however, it has become increasingly clear that it is primarily a neurotransmitter or neuromodulator and that it exerts its functions mainly by local release from nerve endings. VIP plays a hormonal role only when it is released in large amounts from a tumor, with a consequent overflow into the circulation and grossly elevated plasma concentrations of the peptide. Moderately increased VIP plasma and tissue concentrations that cause mainly local effects are found in intestinal ischemia. Crohn's disease and some other chronic inflammatory diseases of the bowel. VIP is also measured in increased amounts in the normal fetus and neonate, where it may play an important physiological role. Such an increase of VIP levels in the circulation could enhance perfusion and metabolic activity of tissues during their rapid-growth period. On the other hand, disorders with a disturbed VIP function such as achalasia and Hirschsprung's disease and possibly also asthma and cystic fibrosis seem to be characterized mainly by a derangement of smooth muscle activity and/or exocrine secretion. Considering this list of disorders where VIP has either a proven or suspected role, it is easy to imagine the significance of this peptide in pediatric pathophysiology. PMID- 2993983 TI - Bicuculline and spinal inhibition produced by dorsal column stimulation in the cat. AB - In barbiturate anaesthetized cats, dorsal column stimulation inhibited ascending volleys recorded in the antero-lateral spinal fasciculus from electrical stimulation of the contralateral tibial nerve and the excitation of neurones of the dorsal horn by noxious heating of the skin. The inhibition was non-selective. Intravenous bicuculline (0.2-0.6 mg/kg) reduced dorsal column induced inhibition of ascending volleys. Bicuculline but not strychnine, administered electrophoretically from micropipettes, reduced dorsal column induced inhibition of the excitation of dorsal horn neurones by noxious heat. These findings suggest that the inhibition studied was produced by release of gamma-aminobutyric acid. This amino acid may play a role in the clinical suppression of pain produced by dorsal column stimulation. PMID- 2993984 TI - A psychophysical analysis of morphine analgesia. AB - Intravenous administration of 0.04-0.08 mg/kg morphine sulfate reduced both sensory intensity and unpleasantness visual analogue scale (VAS) responses to graded 5 sec nociceptive temperature stimuli (45-51 degrees C) in a dose dependent manner. The lower doses of morphine (0.04 and 0.06 mg/kg) resulted in statistically reliable reductions in affective but not sensory intensity VAS responses, possibly reflecting supraspinal effects on brain regions involved in affect and motivation. However, the highest dose of morphine tested (0.08 mg/kg) reduced both sensory and affective VAS responses to graded nociceptive stimuli as well as VAS sensory responses to first and second pain evoked by brief heat pulses. Morphine also had an especially potent inhibitory effect on temporal summation of second pain that is known to occur when intense nociceptive stimuli occur at rates greater than 0.3/sec. The results support current hypotheses about neural mechanisms of narcotic analgesia and further clarify the relative effects of morphine on sensory and affective dimensions of experimental pain. The derived morphine dose-analgesic response functions also provide a reference standard for quantitatively comparing magnitudes of different CNS-mediated forms of analgesia. PMID- 2993985 TI - Spatial summation of pre-pain and pain in human teeth. PMID- 2993986 TI - Responses of the plant parasitic nematodes Rotylenchulus reniformis, Anguina agrostis and Meloidogyne javanica to chemical attractants. AB - Rotylenchulus reniformis, Anguina agrostis and Meloidogyne javanica respond differently to gradients of chemical attractants. In chemotaxis assays performed on agarose plates, R. reniformis L2 larvae oriented their movement to several common inorganic salts, cyclic AMP and AMP, as well as to germinated host plant seeds. M. javanica L2 larvae were attracted to germinated seeds, but not to the salts tested, and A. agrostis dauer larvae were not strongly attracted to any of 12 different tested agents, including host root or shoot tips. Attraction of R. reniformis to salts was measured by comparing different pairwise combinations of ions at equivalent concentrations. The indicated order of attractiveness was: Cl- greater than Na+ greater than C2H3O2- greater than Mg2+, NH4+, SO4(2-). The least attractive salts, (NH4)2 SO4 and MgSO4, were weakly attractive at an orientation threshold of 1 mM, whereas the most attractive salt, MgCl2, was strongly attractive at a threshold of 0.2 mM Cl-. 3',5' cyclic AMP was strongly attractive at a threshold of 0.05 mM, whereas 5'-AMP was a weak attractant. Some of these responses may affect the distribution of R. reniformis in its natural environment. PMID- 2993987 TI - [Effects of aminoglycosides on phosphoinositide metabolism in renal proximal tubules. In vivo and in vitro studies]. AB - Phosphoinositides play a major role in the interaction between aminoglycosides and renal epithelial cells. We therefore investigated both in vivo and in vitro, in isolated proximal tubules and in various parts of the rabbit kidney, the influence of neomycin, gentamicin and amikacin on the phosphoinositide metabolism. This was carried out by measuring the 32p incorporation into phosphatidylinositol 4,5-bisphosphates (PI-P2) and phosphatidylinositol 4 phosphates (PI-P). In aminoglycoside-treated animals, compared to untreated controls, the 32 p-PI-P2 and 32p-PI-P levels were decreased and increased respectively in proximal tubules and in cortical areas but these levels were unchanged in papilla and in medulla. Similar variations could be obtained after exogenous addition of aminoglycosides. PI-P2 is a key membrane component in the regulation of both the Ca2+ transport and the intracellular Ca2+ mobilization. Thus, the aminoglycoside-induced modifications of the phosphoinositide metabolism may be considered as a primary event in the drug toxicity which may result from alterations in the cell calcium homeostasis. PMID- 2993988 TI - Unilateral hypoplasia of the hemithorax causing "pseudoscoliosis" after lung irradiation in a child with Wilms' tumor. AB - A child receiving unilateral pulmonary radiation therapy for metastatic Wilms' tumor in early childhood was noted to develop gradually an asymmetrical thorax. This was clinically diagnosed as scoliosis but radiographs indicated pulmonary hypoplasia as the true cause. PMID- 2993990 TI - Modulation of glucocorticoid secretion by growth hormone. AB - We measured the cortisol and corticosterone responses to insulin-induced hypoglycemia in 13 growth hormone (GH)-deficient children and 30 short children without GH deficiency. Although there was no difference between the two groups in degree of hypoglycemia attained, baseline cortisol, baseline corticosterone, or cortisol 40 min after insulin injection, GH-deficient children had a significantly greater corticosterone response to this stress (3.6 +/- 0.4 versus 1.9 +/- 0.2 micrograms/dl). (All data are presented as mean +/- SEM.) In order to explore the effect of GH on corticosterone secretion, we measured cortisol and corticosterone responses to synthetic (1-24) ACTH before and after 3 days of exogenous GH (0.2 unit/kg/day). In 13 GH-deficient children, GH treatment caused a significant decrease in the corticosterone response to ACTH (2.2 +/- 0.2 micrograms/dl before GH to 1.6 +/- 0.2 micrograms/dl; t = 5.22, p less than 0.001; paired t test) despite the fact that there was no significant change in the cortisol response to ACTH (18 +/- 2 micrograms/dl before and 16 +/- 2 micrograms/dl after). When seven short children who were not GH deficient underwent a similar 3-day course of GH, the decrease in their corticosterone response was much less although still statistically significant (2.0 +/- 0.5 to 1.8 +/- 0.5 micrograms/dl; paired t test, p less than 0.05). Again, the stimulated levels of cortisol were not affected by GH treatment (19 +/- 4 versus 18 +/- 3 micrograms/dl) These results indicate that GH modulates the adrenal response to ACTH by suppressing corticosterone secretion without affecting cortisol secretion. In summary, this study presents two new findings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 2993989 TI - The effects of sodium bicarbonate on brain blood flow and O2 delivery during hypoxemia and acidemia in the piglet. AB - Metabolic acidosis in the neonate is often secondary to hypoxemia and cardiopulmonary disturbances. Sodium bicarbonate, an agent used to treat metabolic acidemia in newborns, is often administered during hypoxemia. In the absence of acid-base alterations, during hypoxemia a reciprocal relationship exists between arterial O2 content (CaO2) and brain blood flow (BBF). However, when hypoxemia is compounded by acidemia it is unclear whether the increase in arterial pH achieved by infusions of sodium bicarbonate alters BBF. To investigate this, BBF (microsphere technique), arterial blood gases, and CaO2 were measured in 14 ventilated piglets. Variables were assessed during a control period, a period of hypoxemia (50 min) associated with metabolic acidemia (hypoxemia + acidemia), and after infusions of either saline (n = 6) or NaHCO3 (n = 8, 2 mEq/kg) during continued hypoxemia. Arterial pH was similar in both groups at control, and hypoxemia + acidemia resulted in comparable reductions of pH in both saline- and NaHCO3-treated piglets (7.21 +/- 0.02 versus 7.21 +/- 0.03, respectively). NaHCO3 infusions produced a significant rise in pH, 7.30 +/- 0.03 versus 7.15 +/- 0.03, p less than 0.05. In each group CaO2 paralleled changes in pH but did not differ between groups. In all animals BBF increased more than 2 fold during hypoxemia + acidemia and was unaltered by infusions of either saline or NaHCO3. Brain O2 delivery decreased in both groups during hypoxemia + acidemia and was unchanged by infusions of saline or NaHCO3. During hypoxemia + acidemia the change in arterial pH induced by NaHCO3 (2 mEq/kg) does not alter BBF or brain O2 delivery. PMID- 2993991 TI - Age-related changes in the adrenergic control of glycogenolysis in rat liver: the significance of changes in receptor density. AB - The present study examined the developmental changes in the adrenergic control of glycogenolysis in the rat model. A relatively new beta-adrenergic radioligand, 125I-iodocyanopindolol (ICP), was examined in binding assays with rat liver plasma membrane (LPM). ICP demonstrated both a higher specificity and a greater affinity for beta-adrenergic receptors than any previously available beta adrenergic radioligand used to study rat LPM. Utilizing this new ligand it was found that beta-adrenergic receptor density decreased from 114 +/- 4 fmol mg-1 in newborn LPM to 19 +/- 3 fmol mg-1 in adult male LPM. In contrast alpha-adrenergic receptor density examined using 3H-prazosin increased from 161 +/- 14 fmol mg-1 in the newborn to 554 +/- 59 fmol mg-1 in the adult male. The results of ICP displacement assays employing various beta-adrenergic agonists and antagonists indicated that ICP binding sites were beta 2-adrenergic receptors. Both guanosine triphosphate and its nonhydrolyzable synthetic analog guanylyl-imidodiphosphate lowered the affinity of epinephrine for ICP binding sites similarly in newborn and adult LPM. Thus the coupling of receptor to guanine nucleotide regulatory protein appeared to be the same in both age groups examined. In isolated hepatocytes glycogen phosphorylase activation was mediated by beta 2-adrenergic stimuli in the newborn and predominantly alpha-adrenergic stimuli in the adult male. These results suggest that the change in glycogen phosphorylase activation from beta- to predominantly alpha-adrenergic mechanisms seen with maturation is related to changes in receptor density. PMID- 2993992 TI - Adenovirus infections in young children. AB - The importance of adenovirus in initiating respiratory disease in young children is stressed in this report. The incidence, clinical illness, asymptomatic carriage, and serologic response of acute adenovirus-associated infection are described in a carefully followed cohort of normal children cultured with each episode of febrile respiratory illness. During a 6-year period, 8.2% of 1,416 nasal washings obtained from sick infants and children less than 7 years of age yielded adenovirus. Adenoviruses were isolated from only 1/174 (0.6%) cultures taken from well children. Typing of 98 isolates showed 81% to be types 1 or 2. A greater than or equal to fourfold rise in neutralizing titer was seen in 45/59 (76%) sampled. In a subset of the cohort observed for 2-week periods in a day care setting, 14 of 21 well children (67%) exposed to symptomatic children with adenovirus infection developed febrile respiratory symptoms and shedding of the same serotype within 2 weeks of exposure. This study confirms that adenovirus has a high attack rate and causes significant respiratory disease in young children. PMID- 2993993 TI - [Importance of determining gastrin, corticotropin and somatotropin in chronic gastroduodenitis in children]. PMID- 2993994 TI - [Total body irradiation by X-ray for adult T-cell leukemia]. PMID- 2993996 TI - On the specificity of the immunoglobulin G produced by chickens experimentally infected with Marek's disease virus. AB - In chickens experimentally infected with Marek's Disease virus (MDV) an increased amount of immunoglobulin G is produced. Using a technique of quantitative crossed immunoelectrophoresis it has been shown, that 70% of this immunoglobulin G is non specific. Only 18% could be absorbed with MDV strain CPRL VII-infected chicken kidney cells, and only 5% with MDV-induced lymphoblastoid cells of the MDCC-MSB1 cell line. It is hypothesized that the production the unspecific immunoglobulin G is caused by a polyclonal stimulation of B-cells. PMID- 2993997 TI - [Rearrangement of the proto-oncogene c-myc in fresh cells from Burkitt's leukemia (L3)]. AB - The presence of a rearrangement of the proto-oncogene c-myc was investigated in the DNA of fresh cells isolated from the blood of two patients with Burkitt's leukemia (L3), and from the node biopsy of a patient with Burkitt's lymphoma. Both samples from the L3 leukemia patients had the characteristic t(8;14) translocation, while the lymphoma specimen presented no abnormality of chromosome 8. Only one of the leukemic DNA's presented a rearranged c-myc pattern, with the breakpoint region located between the first and the second exon. The c-myc pattern of the two other patients appeared normal. The finding of a rearranged c myc oncogene in fresh cells from a Burkitt's leukemia is direct evidence for the implication of c-myc in the disease whereas most of the rearrangements previously described have been found in cell lines established in culture. PMID- 2993995 TI - [Ethmoid tumors in moose and roe deer]. AB - Ethmoid tumors are expansively-infiltratively growing tumors of carcinomatous or sarcomatous nature, deriving from the mucous membrane of the ethmoid bone. In Sweden, such tumors were found in 35 elks (Alces a. alces) and 4 roe deer (Capreolus capreolus) during the years 1947-1982, that means a frequency of about 1 and 0.1 per cent, respectively of the investigation material. However, in the free living elk and roe deer population, the frequency might be much lower. The tumors were malign, extensively melting the soft and hard tissues of the ethmoid region, breaking into the brain cavity, the forehead subcutaneous tissues, etc. Symptoms as suppurative or bloody discharge at the nose, external outline aberrations and disorders to be related to injuries of the central nervous system were observed. In the elk, ethmoid tumors were found only in female animals. In the beginning of this century, ethmoid tumors were found in a number of cattle and horses in Sweden and Norway. Multiple cases occurred in some herds indicating that the tumors were caused by an infectious agent. Since the year 1916, there seem to be no reports on the finding of ethmoid tumors in domestic animals in the Nordic countries. In 1960, however, such tumors were discovered in Indian cattle in Kerala in the south of India. Tumor tissue from the cattle was examined and a herpes-virus was found. Geographically, the distribution of the tumor cases in cattle and elk was very similar in Sweden indicating a possible mutual transmission. As the tumors obviously have disappeared from cattle but not from the elk, it seems likely that the elk might be the primary carrier of the ethmoid tumor. Ethmoid tumors have been observed for many years in Scandinavia but only rather recently they were discovered in India. It has been known for long that birds after contamination might be involved in the spread of virus diseases, provided the virus are reasonably resistant. In the actual case, the suspicion has mainly been directed at three bird species, viz. the blue throat (Luscinia svecica), the scarlet grosbeak (Erythrina erythrina) and the red-necked phalarope (Phalaropus lobatus). In spring and summer, these birds periodically reside in elk habitat where they might be contaminated. In the autumn, they may extend their migration to the southern parts of India. PMID- 2993998 TI - The pattern of expression of the Xenopus laevis tadpole alpha-globin genes and the amino acid sequence of the three major tadpole alpha-globin polypeptides. AB - We have isolated cDNA clones derived from three tadpole alpha-globin mRNAs of Xenopus laevis. The entire nucleotide sequence of the three mRNAs has been determined from the cDNA clones and is presented together with the deduced amino acid sequence of the encoded polypeptides. Two of the three polypeptide sequences are 96% homologous whilst the third sequence is highly diverged, with only a 72% homology. The three tadpole alpha-globin genes are all similarly diverged from the two X. laevis adult alpha-globin genes with which they display approximately 50% homology. Analysis of several independent clones from each class of tadpole alpha-globin sequence reveals a very high degree of coding region polymorphism for each of the three corresponding genes. Using the cloned DNA sequences as hybridisation probes, we have analysed the expression of the corresponding genes during larval development. We show that all three genes are activated simultaneously early in development and that thereafter all three are expressed at an approximately equivalent level. A fourth tadpole alpha-globin mRNA sequence, for which we do not have a cDNA clone, accumulates co-ordinately with the three major mRNA sequences but to a much lower concentration. This pattern of gene expression differs significantly from that of the tadpole beta-globin genes of X. laevis, despite the two classes of genes being closely linked in the genome. PMID- 2994000 TI - Homologies between X and Y chromosomes detected by DNA probes: localisation and evolution. AB - We have isolated and characterized DNA probes that detect homologies between the X and Y chromosomes. Clone St25 is derived from the q13-q22 region of the X chromosome and recognizes a 98% homologous sequence on the Y chromosome. Y specific fragments were present in DNAs from 5 Yq-individuals and from 4 out of 7 XX males analysed. An X linked TaqI RFLP is detected with the St25 probe (33% heterozygosity) which should allow one to establish a linkage map including other polymorphic X-Y homologous sequences in this region and to compare it to a Y chromosome deletion map. Probe DXS31 located in Xp223-pter detects a 80% homologous sequence in the Y chromosome. The latter can be assigned to Yq11-qter outside the region which contains the Y specific satellite sequences. ACT1 and ACT2, the actin sequences present on the X and Y chromosomes respectively, have been cloned. No homology was detected between the X and Y derived fragments outside from the actin sequence. ACT2 and the Y specific sequence corresponding to DXS31 segregate together in a panel of Y chromosomes aberrations, and might be useful markers for the region important for spermatogenesis in Yq. Various primate species were analysed for the presence of sequences homologous to the three probes. Sequences detected by St25 and DXS31 are found only on the X chromosome in cercopithecoidae. The sequences which flank ACT2 detect in the same species autosomal fragments but no male specific fragments. It is suggested that the Y chromosome acquired genetic material from the X chromosome and from autosomes at various times during primate evolution. PMID- 2993999 TI - Nucleotide sequence and mutational analysis of an immunity repressor gene from Bacillus subtilis temperate phage phi 105. AB - We have identified and sequenced a bacteriophage phi 105 gene encoding an immunity repressor, the first to be characterized from a temperate phage infecting a Gram-positive host. Using superinfection immunity as an assay for repressor function, the phi 105 repressor gene was located within a 740-bp PvuII HindIII subfragment near the left end of the phi 105 EcoRI-F fragment. We show that the repressor is specified by the 5'-proximal coding sequence of a translationally overlapping gene pair, transcribed from right to left on the conventional phi 105 map. Comparison of its amino acid sequence (146 residues) with that of a large number of Gram-negative bacterial and phage repressors revealed a putative DNA-binding region between positions 20 and 39. The coding region is preceded by a strong Shine-Dalgarno sequence 5' AAAGGAG 3'. Deletion analysis of the 5'-flanking DNA allowed to identify transcriptional control elements. Their structure, 5' TTGTAT 3' at -35 and 5' TATAAT 3' at -10, strongly suggests that the phi 105 repressor gene is transcribed by the major vegetative form of B. subtilis RNA polymerase, as would be expected for an early phage gene. PMID- 2994001 TI - Inhibition of herpes simplex thymidine kinase gene expression by DNA methylation is an indirect effect. AB - The biological activity of in vitro methylated HSV-TK DNA was analysed after microinjection into thymidine kinase negative rat 2 cells. It was found that the fully methylated DNA (Hpa II methylase) was as active as the non methylated control DNA for about 48 hours after injection. DNA reextraction experiments and blot analysis showed that DNA demethylation was not the reason for the observed TK activity. With prolonged cultivation time the methylated DNA becomes rapidly inactive and 100 hrs after injection thymidine incorporation was no longer detectable in the recipient cells. In transformed cells, obtained after coinjection with SV40 DNA, the HSV-DNA was partially demethylated and inactive. Addition of 5-azacytidine to the culture medium induced further demethylation and reactivation of the thymidine kinase gene. PMID- 2994002 TI - Structure of the mouse glial fibrillary acidic protein gene: implications for the evolution of the intermediate filament multigene family. AB - We report the complete sequence of the gene encoding mouse glial fibrillary acidic protein (GFAP), the intermediate filament (IF) protein specific to astrocytes. The 9.8 kb gene includes nine exons separated by introns ranging in size from 0.2 to 2.5 kb. A comparison of the organization of the GFAP gene with that of genes encoding other IF proteins reveals that the structure of IF genes is highly conserved in spite of considerable divergence at the amino acid level. Thus, most of the evolutionary events leading to the placement of introns in IF genes must have occurred prior to the duplication and subsequent divergence of IF genes from a presumptive common ancestral sequence. The conserved gene organization is unrelated to structural features of IF proteins. A curious feature of the GFAP gene is the large number of repeated sequences found in the introns. Six tracts of reiterated di- or trinucleotides are present, plus tandem repeats of two different novel sequences. One repeat is unique to the GFAP gene; the other occurs elsewhere in the mouse genome, although at relatively low frequency. PMID- 2994003 TI - Interspersed repetitive and tandemly repetitive sequences are differentially represented in extrachromosomal covalently closed circular DNA of human diploid fibroblasts. AB - Extrachromosomal covalently closed circular DNA (cccDNA) was isolated from human diploid fibroblasts by alkaline denaturation/renaturation and CsCl-ethidium bromide isopycnic centrifugation. Probing across these gradient fractions showed a higher proportion of cccDNA sequences homologous to the interspersed highly repetitive Alu I and Kpn I sequences than to the human tandemly-repetitive Eco RI (alphoid) DNA. Cloning of these cccDNAs was then carried out following digestion with restriction endonucleases Hind III, Bam HI or Pst I, and ligation into plasmid pBR322. Many isolated recombinant clones were unstable as seen by a high rate of loss over four cycles of antibiotic selection, and frequent plasmid modifications including deletions adjoining the site of insertion. Of 107 cloned sequences which appeared relatively stable, i.e., survived four cycles of antibiotic selection without incurring detectable deletions, 28% and 11% showed homology to Alu I and Kpn I families, respectively, while 4% contained sequences homologous to both. In contrast, less than one percent hybridized to probes for tandemly-repetitive sequences, Eco RI and Satellite III. The average insert size of cloned cccDNA derived from human fibroblasts, 2.52 Kbp, was larger than previously reported for similar clones derived from genetically less stable permanent lines, which may reflect differences in the process of cccDNA generation. PMID- 2994004 TI - Exon mutations that affect the choice of splice sites used in processing the SV40 late transcripts. AB - The spliced species of late SV40 RNAs present in the cytoplasm of cells infected with various wild-type and mutant strains of SV40 that differ in their leader regions were determined using a novel modification of the primer extension method and the S1 nuclease mapping technique. These data indicated that mutations within the first exon of the late RNAs can affect dramatically the utilization of downstream donor and acceptor splice sites. In one instance, a ten base pair insertion within the predominant first exon increased utilization of an infrequently utilized donor splice site such that the small alteration became part of an intervening sequence, thereby suggesting a novel mechanism for regulation of gene expression. In addition, our method enabled detection of a previously unidentified spliced species, representing less than one percent of the SV40 late 19S RNA present in cells infected with wild-type virus, that may be an intermediate in the synthesis of a known doubly spliced 16S RNA species of SV40. PMID- 2994005 TI - Phylogenetic conservation of immunoglobulin heavy chains: direct comparison of hamster and mouse Cmu genes. AB - We have analyzed the JH-Cmu locus of the Syrian hamster by DNA cloning and sequencing. The single Cmu gene is highly homologous to that of the mouse. The hamster equivalents of the JH and switch (S) recombination regions are arranged as in the mouse, but surprisingly are not highly conserved. Also unlike its close murine relative, the Smu regions among inbred hamster strains are not polymorphic. The complete nucleotide sequence of hamster and mouse Cmu genes have been compared to partial Cmu sequences of other species. Conservation within a portion of the 3' untranslated region may signify functional requirements for 3' end processing. Mutational frequencies within exons and introns of hamster and mouse do not support the theory that the rate of DNA transitions to transversions decreases with evolutionary distance. PMID- 2994006 TI - Diepoxybutane forms a monoadduct with B-form (dG-dC)n.(dG-dC)n and a crosslinked diadduct with the left-handed Z-form. AB - The characteristics of the reactions of DL-diepoxybutane (DEB) with (dG-dC)n.(dG dC)n in the right-handed B-form or the left-handed Z-form were investigated. DEB does react with right-handed B-DNA since less salt is required to convert the modified B-form to Z-form than for the unmodified DNA. However, the product appears to be a monoadduct rather than the crosslinked diadduct formed with the Z form. The modified B-form can be isolated, converted to a Z-form with l mM MnCl2, and then this activated complex further reacts intramolecularly to give the crosslinked Z-product. This modified Z-form cannot be reverted to the B-form unless the crosslink is cleaved with periodate. Only MnCl2, and to a lesser extent ZnCl2, was effective in facilitating the intramolecular conversion of the B-DNA monoadduct to the Z-DNA diadduct; lmM MgCl2 and 4M NaCl were ineffective suggesting that somewhat different types of modified left-handed conformations were generated by the different salts. DEB also cleaves DNA under our reaction conditions thus precluding studies with supercoiled recombinant plasmids harboring segments that adopt Z-structures. PMID- 2994007 TI - The nucleotide sequence of the tnpA gene completes the sequence of the Pseudomonas transposon Tn501. AB - The nucleotide sequence of the gene (tnpA) which codes for the transposase of transposon Tn501 has been determined. It contains an open reading frame for a polypeptide of Mr = 111,500, which terminates within the inverted repeat sequence of the transposon. The reading frame would be transcribed in the same direction as the mercury-resistance genes and the tnpR gene. The amino acid sequence predicted from this reading frame shows 32% identity with that of the transposase of the related transposon Tn3. The C-terminal regions of these two polypeptides show slightly greater homology than the N-terminal regions when conservative amino acid substitutions are considered. With this sequence determination, the nucleotide sequence of Tn501 is fully defined. The main features of the sequence are briefly presented. PMID- 2994008 TI - On the fidelity of DNA polymerase alpha: the influence of alpha-thio dNTPs, Mn2+ and mismatch repair. AB - The phi X174am16 revertant system has been used to investigate the influence of alpha-thio-dNTPs and of Mn2+ on the fidelity of the 9S DNA polymerase alpha from calf thymus. Upon substituting dGTP by alpha-thio-dGTP during the in vitro replication, a nearly tenfold decrease in the frequency of G:G and G:T mispairs is observed. The formation of all other mispairs is not changed in the presence of the corresponding alpha-thio-dNTP. Mn2+ at concentrations of 0.5 mM does not influence the frequencies of the mispairs. The expression rate of errors formed during in vitro replication in the (-) strand has been determined for all mispairs detectable in the phi Xam16 system. The (-) strand expression of G:T, T:T and C:T mismatches is about 50%, whereas for A:G, G:G and C:A mismatches it is clearly below 50%. We conclude that the different base-base mismatches are repaired with different efficiencies. PMID- 2994009 TI - Nucleotide sequence of the Bacillus subtilis xylose isomerase gene: extensive homology between the Bacillus and Escherichia coli enzyme. AB - The xylose isomerase gene from Bacillus subtilis was cloned from a genomic BamH1 library by complementation of an isomerase defective Escherichia coli strain as previously described. The ATG initiation codon is preceded by a Shine-Dalgarno sequence and two hexamers being characteristic for the promoter region of Bacillus genes. The structural gene consists of 1320 base pairs, thus coding for a polypeptide chain of 440 amino acids with a molecular weight of 49 680. The polypeptide primary structure shows over 50% homology to that of the E. coli xylose isomerase. PMID- 2994010 TI - An RFLP detecting single copy X-chromosome fragment, dic56, from Xp22-Xpter [HGM8 assignment no. DXS 143]. PMID- 2994011 TI - Investigation of restriction-modification enzymes from M. varians RFL19 with a new type of specificity toward modification of substrate. AB - The characterization of MvaI restriction-modification enzymes, isolated from Micrococcus varians RFL19, is reported. Both enzymes recognize the 5'CC decreases (A/T)GG nucleotide sequence. The endonuclease cleaves the sequence at the position indicated by the arrow, whereas the methylase modifies the internal cytosine, yielding N4-methylcytosine. This type of modification protects the substrate from R.MvaI cleavage. 5-Methylcytosine in the same position of the recognition sequence does not protect the substrate from R.MvaI cleavage. R.MvaI proved to be the first example of a restriction endonuclease differentiating the position of the methyl group in the heterocyclic ring of cytosine, located in the same site of the recognition sequence. M.MvaI modifies DNA dcm+ in vitro yielding N4,5-dimethylcytosine. N4-methylcytosine cannot be differentiated from cytosine using the Maxam-Gilbert DNA sequencing procedure. PMID- 2994012 TI - Type II restriction endonucleases cleave single-stranded DNAs in general. AB - Restriction endonucleases (13 out of 18 species used for the test) were certified to cleave single-stranded(ss)DNA. Such enzymes as AvaII, HaeII, DdeI, AluI, Sau3AI, AccII,TthHB8I and HapII were newly reported to cleave ssDNA. A model to account for the cleavage of ssDNA by restriction enzymes was proposed with supportive data. The essential part of the model was that restriction enzymes preferentially cleave transiently formed secondary structures (called canonical structures) in ssDNA composed of two recognition sequences with two fold rotational symmetry. This means that a restriction enzyme can cleave ssDNAs in general so far as the DNAs have the sequences of restriction sites for the enzyme, and that the rate of cleavage depends on the stabilities of canonical structures. PMID- 2994013 TI - Cloning of cDNA encoding a 100 kDa nucleolar protein (nucleoline) of Chinese hamster ovary cells. AB - Nucleoline (100 kDa) is the major nucleolar protein in exponentially growing cells that behaves like a nucleolar organizer protein and plays a key role in rDNA transcription and prerRNA processing. We reported the isolation of 5 cDNA clones by probing a cDNA library, constructed in the expression vector lambda gt11, with a polyclonal serum raised against nucleoline. A new immunoassay, using hybrid proteins (beta gal-cDNA encoded protein) was developed to establish that the isolated cDNAs encoded parts of nucleoline. A further confirmation resulted from the sequence comparison between the cDNA encoded peptide and a 42 aa peptide isolated from rat nucleoline (1). The 5 cDNAs overlapped extensively and covered more than 90% of a full length cDNA. By probing a Northern blot with the 100 kDa cDNA, a 2650 nucleotide polyA+ RNA was detected that contained just enough information to code for nucleoline. PMID- 2994014 TI - Infectivity and in vitro mutagenesis of monomeric cDNA clones of citrus exocortis viroid indicates the site of processing of viroid precursors. AB - Monomeric cDNA clones of citrus exocortis viroid (CEV) were constructed in the plasmid vector pSP6-4 and the infectivity of the clones plus in vitro-synthesized RNA transcripts determined by inoculation onto tomato seedlings. Infectivity was dependent on the site of the viroid molecule used for cloning and the orientation of the cDNA insert. Only the plus BamHI cDNA clone was infectious and produced progeny viroid with wild-type sequence at the region corresponding to the BamHI cloning site. Infectivity correlated with the terminal repetition of 11 nucleotides of viroid sequence, 5'GGATCCCCGGG 3', in the vector adjacent to the insert. The 11-nucleotide sequence lies within the highly conserved central region of viroids. Site-directed mutagenesis of a single nucleotide in the repeat at the 5'-end of the CEV insert to 5' GGATCCCC(T,A)GG 3' gave two point mutants. The two mutant CEV inserts, when excised from the vector, were not infectious. However, plasmid DNA and RNA transcripts from non-excised mutant CEV inserts were infectious. The progeny of one of these clones was examined and contained wild type sequence. It was concluded that in vivo processing of longer-than-unit length CEV occurs at one of three adjacent sites in the 11 nucleotide sequence and that the G nucleotide at position 97 is important for viroid replication. PMID- 2994015 TI - Isolation and sequence analysis of a cDNA encoding the ATP/ADP translocator of Zea mays L. AB - A cDNA complementary to the mRNA for the ATP/ADP translocator of maize (Zea mays L.) has been identified by virtue of hybridisation with the homologous gene from yeast. The cloned cDNA has been shown by DNA sequence analysis to contain an open reading frame of 954bp., which encodes a polypeptide of molecular weight 40,519. This polypeptide exhibits a high degree of homology to the translocator polypeptides of beef heart and Neurospora crassa mitochondria. PMID- 2994017 TI - Complete nucleotide sequence of the lipase gene from Staphylococcus hyicus cloned in Staphylococcus carnosus. AB - The lipase gene from Staphylococcus hyicus subsp. hyicus was cloned in Staphylococcus carnosus and Escherichia coli. In both host organisms the lipase gene is expressed and the enzyme is released to the medium. The cloned DNA insert is 2.5 kb in length and DNA sequencing has revealed the location of the gene, the ribosomal binding site and the presence of a typical signal sequence. The open reading frame comprises 1923 nucleotides and gives a preprotein of 641 amino acids with a predicted Mr of 71.382. At the 3' end of the structural gene there are three consecutive stop codons and there is also a transcriptional termination signal. PMID- 2994016 TI - Cloning and characterization of the cDNAs for human and rabbit interleukin-1 precursor. AB - DNA sequence complementary to the mRNA for rabbit interleukin-1 precursor (preIL 1) has been cloned from the cDNA library constructed using partially purified poly(A)+RNA from induced rabbit alveolar macrophages by mRNA hybridization translation assay. By using this cDNA as a probe, human IL-1 cDNA was isolated from the cDNA library prepared using poly(A)+RNA from induced HL-60 cells, a human monocyte-like cell line. The amino acid sequences of the human and rabbit preIL-1 deduced from the cDNA sequences reveal their primary structures which consists of 271 and 267 amino acid residues, respectively. The amino acid sequence is 64% conserved between human and rabbit. The difference in number of amino acid residues results from the carboxy-terminal extention of 4 amino acid residues in human preIL-1. Expression of the cloned human cDNA in E. coli yielded biologically active IL-1. PMID- 2994019 TI - Cloned DNA sequences that determine mRNA stability of bacteriophage phi X174 in vivo are functional. AB - The stability of two species of phi X174 polycistronic mRNA in vivo can be altered by mutating sequences existing immediately upstream of a termination site. The wild type phage contains an mRNA stabilizing sequence ((+) sequence), while the same sequence mutated by insertion ((-) sequence) reduces the stability of the mRNAs. These two sequences were cloned at the 3' ends of gene D or gene B of phi X174 in a pBR322 derivative plasmid. The cloned sequences were functional. The (+) sequence stabilized gene B or gene D mRNA; half-lives of these mRNAs were 7 to 8 min. When the (+) sequence is eliminated ((o) sequence) or replaced with the (-) sequence, the half-lives of the mRNA were reduced to about 1 to 2 min. The stabilization of mRNAs caused an increased production of these proteins. PMID- 2994018 TI - The primary structure of the DeoR repressor from Escherichia coli K-12. AB - The nucleotide sequence of the deoR gene of E. coli, which codes for the DeoR repressor, has been determined. This gene codes for a polypeptide that is 252 amino acids residues in length. Computer-assisted analysis of the nucleotide sequence strongly suggests that the DNA binding domain of the DeoR repressor is located in the N-terminal part of the protein. After the coding region there is a dyad symmetry similar to a palindromic unit present outside many structural genes on the E. coli chromosome. PMID- 2994020 TI - Analysis of the 5' regulatory region of the gene for delta-aminolevulinic acid synthetase of Rhizobium meliloti. AB - Transcriptional regulation of the delta-aminolevulinic acid synthetase gene of Rhizobium meliloti was investigated under conditions of normal vegetative growth and during symbiosis with the legume host alfalfa. S1 nuclease mapping and DNA sequence analysis indicated that transcription originates from two sites separated by 238 base pairs. A deletion analysis of the putative promoter regions P1 and P2, corresponding to the proximal and distal RNA start sites, was carried out with Bal-31 nuclease. Promoter function was monitored as beta-galactosidase activity after fusing the deletions to lac Z and introducing them into Rhizobium on a broad host range plasmid. The data obtained suggest that both regions function equivalently as promoters. The DNA sequences of P1 and P2 show considerable homology in the region between -35 and the start of transcription. Both contain a -35 region that is analogous to the consensus E. coli promoter sequence, while the -10 region is dissimilar. No resemblance was found between either P1 or P2 and the promoter regions of genes under general nitrogen control. PMID- 2994022 TI - Infection risks for nurses. PMID- 2994021 TI - Mapping and 5' end determination of kinetoplast maxicircle gene transcripts from Leishmania tarentolae. AB - Transcripts for six Leishmania tarentolae maxicircle structural genes (cytochrome oxidase subunits I, II and III, cytochrome b, human mitochondrial unidentified reading frames 4 and 5) and several unidentified open reading frames were mapped, and the locations of the 5' ends determined by primer runoff analysis. All genes studied here are transcribed from the same strand as the 12S and 9S ribosomal RNAs except for the cytochrome oxidase subunit I gene. In two cases (ORF3 and ORF4, ORF5 and ORF6), a single transcript covers two contiguous overlapping reading frames. The 5' ends of the RNAs are located 20-64 nt from the putative translation initiation codons. Primary transcripts from a mitochondrial RNA preparation were 5' end-labeled with guanylyltransferase and alpha -32P-GTP; the major labeled species comigrated with the 12S and 9S mitochondrial rRNAs, and in addition there were at least four higher molecular weight labeled species. PMID- 2994023 TI - Peptide binding to intestinal epithelium: distinct sites for insulin, EGF and VIP. AB - Several peptides, including insulin, epidermal growth factor and vasoactive intestinal polypeptide bind to intestinal epithelial cells. However, it is unclear whether one binding site binds several peptides or whether separate sites exist for each peptide. These studies were designed to examine the specificity of peptide binding sites on intestinal epithelial cells. Peptide binding was measured directly with [125I]radiolabelled peptides to isolated enterocytes prepared from rabbit ileum. The characteristics of insulin and epidermal growth factor binding were similar. Both insulin and epidermal growth factor specific binding was saturable, directly correlated to cell concentration and temperature and pH dependent. The total number of insulin binding sites per cell was 4500, that for epidermal growth factor was 2280. Scatchard analysis for both peptides produced curvilinear plots. Dissociation of both peptides from the binding site was increased in the presence of their respective unlabelled peptide. However, insulin specific binding was not altered by epidermal growth factor, and epidermal growth factor specific binding was unaffected by insulin. Further, both insulin and epidermal growth factor failed to inhibit the specific binding of vasoactive intestinal polypeptide to ileal enterocytes, and vasoactive intestinal polypeptide did not inhibit insulin or epidermal growth factor specific binding. These studies demonstrate that insulin, epidermal growth factor and vasoactive intestinal polypeptide interact with three distinct membrane binding sites on the enterocyte. PMID- 2994024 TI - Behavioral support for an ACTH receptor in the CNS. AB - In this report we present a series of experiments which have led us to support the notion of the presence of an ACTH receptor in the CNS. A short intense heat stress (hot-plate) applied to the paws of rats will temporarily reduce activity. During the course of experimentation we were able to eliminate a number of logical mediators. Neither adrenalectomy, adrenal-medullectomy, naloxone administration, nor alpha-MSH-(1-12) were able to affect the observed akinesia. Hypophysectomy, however, was able to abolish or mask the behavior and the reduction in activity could be reinstated by the administration of ACTH-(4-10) to hypophysectomised rats. These data support the notion that a short intense stressor can release ACTH and that this ACTH can be responsible for mediating the short term reduction in activity. In addition, the fact that ACTH-(4-10) has only minimal steroidogenic properties and was able to reinstate the behavior led us to speculate that these effects were of central origin. Furthermore, since naloxone was not capable of altering the behavior, the suggestion is that ACTH in this paradigm acted at a receptor site apart from the naloxone sensitive receptor. This site may in fact be an ACTH specific receptor. PMID- 2994025 TI - Distinct binding sites for substance P and neurokinin A (substance K): co transmitters in rat brain. AB - The distribution of binding sites in rat brain for iodinated neurokinin A and iodinated substance P were compared using autoradiography. Distinct patterns of binding for the two iodinated tachykinins were noted. Binding sites for iodinated neurokinin A were noted in the olfactory bulb, cortex, supraoptic n., paraventricular n., certain amygdaloid n., hippocampus, medial habenula, interpeduncular n., n. of the tractus solitarius, and dorsal horn of the spinal cord. This pattern was in contrast to low levels of binding of iodinated substance P to the cortex, supraoptic n., paraventricular n., and the interpeduncular n., but substantial density of binding sites in numerous other regions. PMID- 2994026 TI - Recent advances in the morphology of myositis. AB - Myositis in man may be divided into infectious and non-infectious forms. The myopathologist more often deals with the latter forms which comprise dermatomyositis/polymyositis, inclusion body myositis, mixed connective tissue disease/collagenoses, and granulomatous myopathies. Modern morphological techniques as enzyme-histochemistry, electron microscopy, immunohistology, and morphometry are of different value in various forms of myositis, but are often indispensable techniques in up-to-date diagnostic work up of a myositis. PMID- 2994027 TI - Ultrastructural characteristics and variations in human mucinous breast carcinomas. AB - Nineteen human mucinous breast carcinomas have been studied by electron microscopy in order to investigate the variations in the fine structure of this type of tumor. All the investigated tumors are characterized by a well-developed, rough endoplasmic reticulum and prominent Golgi complexes. The majority of tumor cells contain secretory granules. Approximately 50% (9/19) of the tumors have cytoplasmic dense core granules that are morphological identical to the neurosecretory granules found in APUD-cell derived tumors. Six out of the nine tumors react positively in a Grimelius staining for light microscopic argyrophilic granules. Two of the investigated breast carcinomas contain tonofilaments that are normally regarded as characteristic of squamous epithelium. It is concluded that mucinous breast carcinomas--that to this time have been regarded as a morphological homogenous group of tumors--are ultrastructurally characterized by heterogeneity. PMID- 2994028 TI - What's new in endocrine factors of tumor growth? AB - In this review some aspects of endocrinological factors in cancer are discussed. The first part addresses the determination of steroid hormone receptors in cancer of the breast. The importance of steroid hormone receptor determination for the clinical management of breast cancer patients is emphasized. Current methods for receptor measurement are critically evaluated and it is concluded that rigorous standardization of tissue handling procedures and of radiochemical methodology is mandatory to improve the reliability of the presently available methods. The development of new techniques for receptor determination has been met with high expectations. Monoclonal antireceptor antibodies in principle allow the development of a radioimmunoassay but are not widely available as yet. Histochemical methods for receptor localization have so far failed to meet accepted criteria of specificity. Therefore radiochemical determination at present remains the only reliable method. The second part addresses the production of peptide hormones by endocrine tumours of the gastroenteropancreatic (GEP) system and by non-neuroendocrine neoplasms. In the endocrine tumours of the GEP system a wide range of peptide hormones can be detected by immunocytochemistry, but most frequently the pattern of hormone production is comparable with that in the normal organ of origin. Clinical symptoms of excess hormone production occur much less frequently than hormone production is found by immunological methods and is usually caused by one hormone. It is proposed that these neoplasms are classified according to the hormone which causes the leading clinical symptoms and/or the hormone with the highest serum level. Peptide hormone producing cells in classical carcinoma are an intriguing phenomenon which has shed some new light on the embryological origin of neuroendocrine cells in the relevant organs and also on the histogenesis of neuroendocrine as well as non neuroendocrine neoplasms. PMID- 2994029 TI - A search for new antihypertensive agents. I. Alpha-adrenergic activity and selection of structures among pyrazine substituted imidazolines. AB - A group of new pyrazinylimidazolines was prepared and their effect on the contraction of isolated rat tail artery as well as their effect on blood pressure in urethane anesthetized normotensive rats were tested. The biological responses were compared to those elicited by standard imidazoline drugs and structure activity relationships were analysed. The new chemical derivatives were subsequently synthetized and tested pharmacologically. Basing on the experimental results obtained and comparing these to the reported data on arylimidazolines the following suggestions have been put forward concerning the further modifications of alpha-adrenergically active pyrazinylimidazolines: 1) a distance of about 5 A should separate imidazoline nitrogen and pyrazine nucleus, 2) bond refractivity of the separating bridge should be above 5.33, 3) hydrophobicity of substituents in pyrazine ring must be possibly high. PMID- 2994030 TI - [Role of adenine nucleotides in the regulation of the structure and properties of erythrocytes]. PMID- 2994031 TI - Diverticular disease. Current trends in therapy. AB - Colonic diverticulosis is a common problem in the United States, apparently related to low dietary intake of fiber. Diverticula may bleed or cause local infection or perforation. When antibiotic treatment of infection fails, surgical intervention is necessary. Currently, the most common procedures are primary resection and anastomosis for limited disease and a two-stage Hartmann procedure for severe disease or abscess. If the fiber hypothesis of diverticular formation is correct, then addition of fiber to the diet should prevent diverticular disease. PMID- 2994032 TI - Glucagonoma without cutaneous manifestations. AB - A 63 year old man presented with features of the glucagonoma syndrome, that is thromboembolic disease, weight loss, raised sedimentation rate, diabetes mellitus, hypoproteinaemia and reduced plasma amino acid levels, but without necrolytic migratory erythema. The plasma glucagon level was raised and the tumour was demonstrated by abdominal CT scan. Immunofluorescent studies of the resected tumour confirmed the diagnosis. The normal tissue zinc status supports the view that necrolytic migratory erythema is related to zinc deficiency. PMID- 2994033 TI - Myocarditis and fulminant hepatic failure from coxsackievirus B infection. AB - A case of fulminant hepatic failure in association with myocarditis is reported. Presentation suggested an acute hepatitis which was complicated by cardiac failure. Evidence of severe myocarditis was found at autopsy. PMID- 2994034 TI - The large scale purification of ubiquitin from human erythrocytes. AB - A simple, reproducible method for the large-scale purification of active ubiquitin from human erythrocytes is described. Erythrocytes contain 100 micrograms free ubiquitin per cc of packed cells, of which 44% can be recovered in homogeneous form by a combination of heat treatment, ammonium sulfate fractionation, and ion exchange chromatography. PMID- 2994035 TI - High-affinity-receptor-mediated uptake and degradation of glucose-modified proteins: a potential mechanism for the removal of senescent macromolecules. AB - Proteins that have been modified by long-term exposure to glucose accumulate advanced glycosylation end products (AGE) as a function of protein age. In these studies, we have characterized the interaction of AGE-protein with mouse peritoneal macrophages, using AGE-modified bovine serum albumin (AGE-BSA, prepared by incubation with glucose) as a probe. AGE-BSA was specifically bound to cells at 4 degrees C and was taken up and degraded at 37 degrees C; these processes were concentration dependent and saturable. Competition experiments with AGE-BSA, BSA incubated with phosphate-buffered saline rather than glucose, and yeast mannan demonstrated that macrophages specifically recognize AGE on proteins by a receptor that is completely distinct from the mannose/fucose receptor. Scatchard analysis of AGE-BSA binding data indicated that there are approximately 1.06 X 10(5) receptors per macrophage, with an affinity constant of 1.75 X 10(-11) M. Specific binding of AGE-BSA to the macrophage receptor was competitively inhibited by BSA that had been chemically coupled to a synthetic analogue of the specific AGE, 2-(2-furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI BSA). FFI-BSA was also taken up by macrophages in a concentration-dependent, saturable manner. Prior incubation of macrophages with AGE-BSA failed to influence the subsequent uptake and degradation of added AGE-BSA. Thus, the AGE receptor does not appear to be down-regulated by exposure to AGE-proteins. Results from these studies suggest that AGE could act in vivo as a specific signal for recognition and degradation of senescent macromolecules. Incomplete removal of AGE-proteins by macrophages may ultimately give rise to some of the physiologic changes that occur with normal aging. PMID- 2994036 TI - Reversibility of arginine-specific mono(ADP-ribosyl)ation: identification in erythrocytes of an ADP-ribose-L-arginine cleavage enzyme. AB - Enzymes have been identified in animal tissues that catalyze the mono(ADP ribosyl)ation of arginine and proteins. Since these NAD:arginine ADP ribosyltransferases under physiological conditions do not appear to catalyze the degradation of the product ADP-ribose-arginine, the possibility was investigated that a different family of enzymes exists that cleaves the ADP-ribose-arginine linkage. An enzyme was identified in and partially purified from turkey erythrocytes that catalyzed the degradation of ADP-ribose-[14C]arginine synthesized by a salt-activated NAD:arginine ADP-ribosyl-transferase, resulting in the release of a radiolabeled compound that was characterized chromatographically and by amino acid analysis as arginine. This putative arginine product was converted in a reaction dependent on NAD and the NAD:arginine ADP-ribosyltransferase to a compound exhibiting properties characteristic of ADP-ribose-arginine. Action of cleavage enzyme on [adenine-U 14C]ADP-ribose-arginine resulted in the release of a radiolabeled compound that behaved chromatographically like [adenine-U-14C]ADP-ribose. Since degradation of ADP-ribose-arginine appears to generate an arginine moiety that is a substrate for the NAD:arginine ADP-ribosyltransferase, it appears that ADP-ribosylation may be a reversible modification of proteins. PMID- 2994037 TI - Stoichiometric methylation of calcineurin by protein carboxyl O-methyltransferase and its effects on calmodulin-stimulated phosphatase activity. AB - Calcineurin, a calmodulin-stimulated protein phosphatase, was a substrate for purified bovine brain protein carboxyl O-methyltransferase (protein O methyltransferase; EC 2.1.1.24) and incorporated up to 2 mol of CH3 per mol of calcineurin. Carboxyl methylation was dependent on the concentrations of S adenosyl-L-[methyl-3H]methionine and was prevented by addition of the carboxyl methylation inhibitor S-adenosylhomocysteine. The stoichiometry of methyl group incorporation was related to the ratio of methyltransferase/calcineurin. The rate of spontaneous hydrolysis of carboxyl methylester groups on calcineurin increased rapidly above pH 6.5 with those on native carboxyl-methylated calcineurin substantially more labile than for trichloracetic acid-precipitated calcineurin. Polyacrylamide gel electrophoresis in the presence of NaDodSO4 (pH 2.4) confirmed that the A subunit of calcineurin (Mr = 61,000) was the primary site of carboxyl methylation with little, if any, modification of the B subunit (Mr = 18,000). When carboxyl-methylated calcineurin (approximately 1-2 mol of CH3 per mol of protein) was assayed for p-nitrophenyl phosphatase activity at pH 6.5, a marked inhibition of calmodulin-stimulated activity was observed while there was little effect on Mn2+-stimulated phosphatase activity. Thus, calcineurin appears to be an excellent substrate for protein carboxyl O-methylation and this modification, which impairs calmodulin stimulation of phosphatase activity, may be of functional significance. PMID- 2994038 TI - Ionic inhibition of formation of RecA nucleoprotein networks blocks homologous pairing. AB - Conditions that favor the complete coating of single-stranded DNA by RecA protein promote the association of these presynaptic filaments with naked double-stranded DNA to form large nucleoprotein networks before homologous pairing occurs. These RecA nucleoprotein networks sequester virtually all of the DNA in the reaction mixture. Conditions that are suboptimal for the formation of the RecA presynaptic filament rendered both the formation of RecA-DNA networks and the subsequent formation of joint molecules sensitive to inhibition by excess ATP or by pyrophosphate when these were added during synapsis. The rate of homologous pairing was directly related to the degree of inhibition of network formation. Various multivalent cations added during synapsis restored both the formation of networks and the pairing of homologous molecules. These observations support the view that the nucleoprotein network is a synaptic intermediate by means of which RecA protein facilitates the conjunction of DNA molecules and the subsequent processive search for homology. Inhibition by multivalent anions and restoration by multivalent cations suggests in addition, that negative charge repulsion inhibits the binding of naked duplex DNA to presynaptic filaments. PMID- 2994039 TI - Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. AB - DDT1 MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain alpha 1-adrenergic receptors (54,800 +/- 2700 sites per cell) that are coupled to stimulation of inositol phospholipid metabolism. Incubation of these cells with tumor-promoting phorbol esters, which stimulate calcium- and phospholipid-dependent protein kinase, leads to a marked attenuation of the ability of alpha 1-receptor agonists such as norepinephrine to stimulate the turnover of inositol phospholipids. This turnover was measured by determining the 32P content of phosphatidylinositol and phosphatidic acid after prelabeling of the cellular ATP pool with 32Pi. These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular alpha 1 receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the alpha 1 receptors were purified approximately equal to 300-fold from control and phorbol ester-treated 32Pi-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the Mr 80,000 alpha 1 receptor ligand-binding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of alpha 1 receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of alpha 1 receptor. The effect of phorbol esters on norepinephrine-stimulated inositol phospholipid turnover and alpha 1-receptor phosphorylation showed the same rapid time course with a t1/2 less than 2 min. These results indicate that calcium- and phospholipid-dependent protein kinase may play an important role in regulating the function of receptors that are coupled to the inositol phospholipid cycle by phosphorylating and deactivating them. PMID- 2994040 TI - [4Fe-4S]-cluster-depleted Azotobacter vinelandii ferredoxin I: a new 3Fe iron sulfur protein. AB - Fe(CN)6(-3) oxidation of the aerobically isolated 7Fe Azotobacter vinelandii ferredoxin I, (7Fe)FdI, is a degradative reaction. Destruction of the [4Fe-4S] cluster occurs first, followed by destruction of the [3Fe-3S] cluster. At a Fe(CN)6(-3)/(7Fe)FdI concentration ratio of 20, the product is a mixture of apoprotein and protein containing only a [3Fe-3S] cluster, (3Fe)FdI. This protein mixture, after partial purification, has been characterized by absorption, CD, magnetic CD, and EPR and Fe x-ray absorption spectroscopies. EPR and magnetic CD spectra provide strong evidence that the [3Fe-3S] cluster in (3Fe)FdI is essentially identical in structure to that in (7Fe)FdI. Analysis of the extended x-ray absorption fine structure (EXAFS) of (3Fe)FdI finds Fe scattering at an average Fe...Fe distance of approximately equal to 2.7 A. The structure of the oxidized [3Fe-3S] cluster in solutions of oxidized (3Fe)FdI, and, by extension, of oxidized (7Fe)FdI, is thus different from that obtained by x-ray crystallography on oxidized (7Fe)FdI. Possible interpretations of this result are discussed. PMID- 2994041 TI - Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma. AB - A complete amino acid sequence has been determined for the UP1 single-stranded DNA binding protein from calf thymus that was first described by G. Herrick and B. M. Alberts [(1976) J. Biol. Chem. 251, 2124-2132]. Peptides required to establish the UP1 sequence were isolated by reversed-phase HPLC of digests produced by endoproteinase Lys-C, trypsin, chymotrypsin, Staphylococcus aureus V8 protease, and cyanogen bromide cleavage of UP1. The purified peptides were coupled to aminopolystyrene prior to solid-phase sequencing. UP1 contains 195 amino acids and has a molecular weight of 22,162. UP1 has a blocked NH2 terminus and contains a single NG,NG-dimethylarginine residue near its COOH terminus. Gas phase sequencing of tryptic peptides derived from an analogous protein from mouse myeloma cells [Planck, S. R. & Wilson, S. H. (1980) J. Biol. Chem. 255, 11547 11556] revealed that this mouse helix-destabilizing protein shares a high degree of sequence homology with UP1. Of the 59 amino acids in the mouse protein that have so far been found to be homologous with UP1, 48 correspond exactly to sequences found in UP1. Most of the 11 differences that have been found between the sequences of these two proteins are conservative in nature, involving primarily the interchange of chemically similar amino acids. One 9-residue mouse sequence that is not obviously homologous to UP1 may be a result of the larger size of the mouse myeloma protein as compared to UP1. Since none of the UP1 or mouse myeloma helix-destabilizing protein sequence appears to be homologous to that of any previously sequenced protein, we presume that these two proteins represent a distinct class of single-stranded nucleic acid binding proteins that probably play a role in metabolism of single-stranded RNA or DNA in vivo. PMID- 2994042 TI - A 5' spliced leader is added in trans to both alpha- and beta-tubulin transcripts in Trypanosoma brucei. AB - The approximately 15 alpha- and 15 beta-tubulin genes of Trypanosoma brucei are arranged in a tandem array of alternating alpha- and beta-tubulin genes. We have examined the structure of mRNA transcripts from the tubulin gene family and have found at the 5' ends of both alpha- and beta-tubulin messages a 35-nucleotide spliced leader (SL) identical to that identified previously on the 5' ends of variant surface glycoprotein (VSG) mRNAs. No 35-nucleotide SL sequences were encoded in the tubulin intergenic regions; instead, the SL sequence apparently originates from small RNA transcripts encoded at sites that are unlinked to the tubulin genes. The tandem arrangement of the tubulin genes and the presence of the SL at the 5' end of both alpha- and beta-tubulin transcripts establishes that RNA joining between the SL and tubulin mRNA occurs in trans. PMID- 2994043 TI - Molecular cloning and amino acid sequence of the precursor form of bovine adrenodoxin: evidence for a previously unidentified COOH-terminal peptide. AB - Several recombinant cDNA clones specific for the mitochondrial iron-sulfur protein adrenodoxin have been identified in a bovine adrenocortical cDNA library. One clone (pBAdx4) contains a 900-base-pair insert that includes the entire amino acid coding region of the adrenodoxin precursor protein. The amino acid sequence of mature adrenodoxin deduced from the nucleotide sequence of pBAdx4 is identical with that determined by protein sequencing except for three amide changes. The previously undetermined sequence of the adrenodoxin NH2-terminal precursor segment (58 amino acids) contains several basic residues, a characteristic feature of the precursor segment of proteins destined for mitochondria. In addition, a 14 amino acid extension is present at the COOH terminus of the mature adrenodoxin sequence. Whether this represents a COOH-terminal precursor segment is not clear. Three different adrenodoxin mRNAs are present [1.75, 1.4, and 0.95 kilobase(s) long] in bovine adrenocortical RNA. RNA from bovine corpus luteum, liver, and kidney contains transcripts that hybridize to adrenodoxin cDNA. Accumulation of adrenodoxin mRNA occurs in cultured bovine adrenocortical cells after treatment with ACTH or dibutyryl-cAMP, similar to that observed for the mitochondrial steroid hydroxylases that it services--namely, the cholesterol side chain-cleavage cytochrome P-450 and the steroid 11 beta-hydroxylase cytochrome P 450. PMID- 2994044 TI - In vitro replication of duplex circular DNA containing the simian virus 40 DNA origin site. AB - Extracts (0.2 M NaCl) of HeLa cells support replication of DNA containing simian virus 40 (SV40) origin in the presence of SV40 large tumor (T) antigen. The reaction leads to the accumulation of high molecular weight products that represent DNA containing one parental strand and one progeny strand as well as duplex molecules that contain both strands derived from the input deoxynucleoside triphosphates. The replication reaction is inhibited by aphidicolin and by camptothecin, two inhibitors known to inhibit eukaryotic DNA replication in vivo. PMID- 2994045 TI - Demonstration of a stimulatory protein for virus-specified DNA polymerase in phorbol ester-treated Epstein-Barr virus-carrying cells. AB - A heat-labile Epstein-Barr virus-specific DNA polymerase stimulatory protein having a molecular mass of 45 kDa was purified from phorbol 12-myristate 13 acetate-treated P3HR-1 cells by column chromatography. The virus DNA polymerase stimulatory protein was precipitated by sera from patients with nasopharyngeal carcinoma but not by sera from healthy donors. The interaction of the stimulatory protein with DNA polymerase was stoichiometric. Furthermore, this protein stimulated Epstein-Barr virus DNA polymerase but not herpes simplex virus type 1 or type 2 or human DNA polymerase alpha. The stimulatory protein did not alter the Km value of dTTP or DNA but did increase the Vmax of DNA polymerase. Salt concentrations between 100 mM and 150 mM KCl were optimal for this protein induced stimulation of Epstein-Barr virus DNA polymerase activity. The presence of the stimulatory protein in the reaction mixture enhanced the sensitivity of virus DNA polymerase to phosphonoformate. PMID- 2994046 TI - Long terminal repeat sequences impart hematopoietic transformation properties to the myeloproliferative sarcoma virus. AB - The myeloproliferative sarcoma virus not only transforms fibroblasts but also causes extensive expansion of the hematopoietic stem cell compartment on infection of adult mice. Similar to the Moloney sarcoma virus, it carries the mos oncogene. Moloney sarcoma virus, however, does not induce myeloproliferation and leukemia in adult mice. The difference between the two viruses was explored by using their molecularly cloned genomes and the cellular mos oncogene to construct recombinant genomes. It was shown that the U3 region of the viral long terminal repeat (LTR) has a decisive function in determining the target cell specificity of the myeloproliferative sarcoma virus. Any mos gene, whether of cellular or viral origin, is sufficient in conjunction with the proper LTR to induce myeloproliferation. Our results indicate that the pathogenicity of acutely transforming viruses is determined not only by the oncogene but also by sequences in the viral LTR. PMID- 2994047 TI - beta-Endorphin: surface binding and internalization in thymoma cells. AB - The opioid peptide beta-endorphin binds to specific nonopioid binding sites (Mr 72,000) that are present on the surface of thymoma cells. beta-Endorphin is then internalized, apparently via the Mr 72,000 species, and is subsequently found within intracellular, vesicular structures. This process is accompanied by the down-regulation of the Mr 72,000 binding sites. Our findings suggest that beta endorphin may modulate cellular functions, such as T-lymphocyte proliferation, at intracellular rather than cell surface sites. PMID- 2994048 TI - Cellular receptor for 125I-labeled tumor necrosis factor: specific binding, affinity labeling, and relationship to sensitivity. AB - Tumor necrosis factor (TNF) is a proteinaceous toxin shed by stimulated myeloid cells. Murine TNF was radioiodinated to a specific activity of 1 mCi/nmol (1 Ci = 37 GBq) of monomer. 125I-labeled TNF (125I-TNF) retained complete cytotoxic activity and it was immunochemically identical to the native toxin in a quantitative immunoprecipitation assay. It could be shown by competition binding that 125I-TNF bound to intact L929 cells with a specificity equal to that of native toxin. The conditions of time, temperature, and concentration involved in equilibrium specific binding to intact cells were studied in detail. When binding was carried out at 4 degrees C for 18 hr, four cell lines sensitive to 125I-TNF cytotoxicity demonstrated high-affinity binding. The binding reached half-maximal level at 3 pM and saturated at 30 pM. These concentrations approximated those required for cell death. Scatchard analysis gave approximately 1000 sites per cell. J774.1 cells, the source of the toxin, demonstrated similar binding but were not sensitive to 125I-TNF cytotoxicity. Other sensitive cell lines and freshly extracted tumor cells showed specific binding at 3 pM. Normal lymphoid organ cell suspensions and two human tumorigenic cell lines were not sensitive and failed to demonstrate specific binding. 125I-TNF, covalently cross-linked to its receptor on sensitive L-M cells with disuccinimidyl suberate, was isolated and analyzed by sodium dodecyl sulfate/polyacrylamide gel electrophoresis and autoradiography. Two specific bands were identified. The most prominent band had a mobility corresponding to a molecular mass of 95 kDa and the second band had a molecular mass of 75 kDa. The presence of the binding site appears to be necessary but not sufficient to explain the sensitivity of cells to the cytotoxic action of TNF. PMID- 2994049 TI - Comparative statistics for DNA and protein sequences: single sequence analysis. AB - Four categories of data representations are used to help interpret structures and similarities of nucleic acid and protein sequences. Statistical significance of the observed relationships revealed by these representations are assessed by a hierarchy of permutation procedures and by comparisons with theoretical random models. Applications are presented for various DNA sequences including papovaviruses, Epstein-Barr virus, mitochondrial genomes, and several globin and immunoglobulin genes. PMID- 2994050 TI - Nucleotide sequence and predicted amino acid sequence of a protein encoded in a small herpes simplex virus DNA fragment capable of trans-inducing alpha genes. AB - The five alpha genes of herpes simplex virus 1 are the first set of genes to be expressed after infection. Previous studies have shown that alpha genes resident in eukaryotic cells are induced by infection with herpes simplex virus 1 or 2 but not by other herpesviruses and indicate that the alpha trans-inducing factor was a structural component of the virion. This factor induces genes linked to a bona fide promoter and containing at the 5' end a small sequence derived from the promoter-regulatory domains of alpha genes. We report the sequence of a small DNA fragment shown previously to be capable of expressing the alpha trans-inducing factor in transient expression systems. The only gene encoded in its entirety in this fragment is predicted to specify a 479 amino acid protein with a Mr of 53,053. The precise termini of the 1.74-kilobase mRNA specifying this protein were determined in our 5' and 3' S1 nuclease protection studies. PMID- 2994051 TI - Glucocorticoid regulation of mouse mammary tumor virus sequences in transgenic mice. AB - We have introduced a chimeric plasmid, pLTR2TK, containing the mouse mammary tumor virus (MTV) long terminal repeat (LTR) linked to the herpes simplex virus type 1 thymidine kinase gene into the mouse germ line by microinjection. In one mouse line, the thymidine kinase gene is appropriately expressed in the lactating mammary glands of heterozygous females; expression also occurs in the ovaries of these mice. In heterozygous males of this line, and in a male derived from another microinjection, transcription of these new germ line genes occurs in the testes and is specifically stimulated by glucocorticoid hormones. Thus, the MTV LTR may contain elements that allow its expression both in lactating mammary glands and in testicular tissue. PMID- 2994052 TI - Anti-immunoglobulin augments the B-cell antigen-presentation function independently of internalization of receptor-antigen complex. AB - All mouse splenic B cells, including small resting B cells, process and present the native globular protein antigens, pigeon and tobacco hornworm moth cytochromes c, to a cytochrome c-specific T-cell hybrid in a major histocompatibility complex-restricted fashion, in the micromolar to nanomolar antigen-concentration range. As is the case for macrophages, treatment with paraformaldehyde or the lysosomotropic agents chloroquine and ammonium chloride blocked processing of the native pigeon protein but did not affect the presentation of a carboxyl-terminal peptide fragment of pigeon cytochrome c (residues 81-104) which contained the T-cell antigenic determinant. However, in contrast to macrophages, whose antigen-processing and -presentation functions are insensitive to radiation, radiation blocked the processing of the native protein but not the presentation of the peptide fragment. The processing and presentation function of the B cells was augmented by F(ab')2 of rabbit anti-mouse Ig antibodies, in that 1/10th to 1/30th as many cells and 1/10th as much antigen were required to maximally activate the T-cell hybrid. This augmentation did not appear to be due to either crosslinking of the Ig receptors or to B-cell activation, as monovalent Fab fragments were nearly as effective as the bivalent reagent, and the concentrations of F(ab')2 anti-Ig used did not induce measurable proliferative responses. Furthermore, enhancement can occur in the absence of cytochrome c binding and internalization, since B cells that were fixed with paraformaldehyde after treatment with F(ab')2 anti-Ig were more effective in presenting the carboxyl-terminal peptide than were untreated fixed cells. The same phenomenon followed the binding of an irrelevant antigen (carboxydinitrophenylated bovine serum albumin) by antigen-binding B cells, resulting in enhanced processing and/or presentation of native pigeon cytochrome c. Thus, nonspecific enhancement of antigen processing and presentation can be obtained by either antigen or anti-Ig binding to the B-cell antigen receptor, both treatments presumably delivering the same signal without requiring internalization of the specifically bound antigen for subsequent processing. PMID- 2994053 TI - Prazosin, an alpha 1-adrenergic receptor antagonist, suppresses experimental autoimmune encephalomyelitis in the Lewis rat. AB - Prazosin, an antagonist of alpha 1-adrenergic receptors, has been found to suppress the clinical and histological expression of experimental autoimmune encephalomyelitis (EAE) in the Lewis rat. Suppression was more significant in females than in males and was a dose-dependent phenomenon. Analysis of the effect of other adrenergic receptor antagonists supports the conclusion that the suppressive effect of prazosin is a consequence of blockade of the alpha 1 receptor since treatment with either the alpha 2-antagonist yohimbine or the beta antagonist propranolol exacerbated the disease, whereas treatment with the long acting mixed alpha 1/alpha 2-antagonist phenoxybenzamine had some suppressive activity. Treatment with prazosin was also able to suppress clinical and histological signs of EAE in animals sensitized by adoptive transfer with activated spleen or lymph node cells. Whether prazosin acts through altering vascular permeability or the immune response, or both, remains to be determined. PMID- 2994054 TI - Enkephalins have a direct positive inotropic effect on cultured cardiac myocytes. AB - Enkephalins have peripheral vascular effects, and enkephalinergic innervation of the heart has been reported. To determine whether enkephalins have direct effects on myocardium, we studied the effects of [D-Ala2, Met5]enkephalinamide and [D Ala2, D-Leu5]enkephalin on amplitude of contraction (measured with an optical video system) in spontaneously beating monolayer cultures of chicken embryo ventricular cells, a preparation devoid of intact neural elements. [D-Ala2, Met5]enkephalinamide and [D-Ala2, D-Leu5]enkephalin as well as [Met5]- and [Leu5]enkephalin increased contractility in a concentration-dependent manner. The enkephalin-induced maximal contractile effects were 28% and 30% above control, with EC50 values of 0.53 and 0.17 microM for [D-Ala2, Met5]enkephalinamide and [D Ala2, D-Leu5]enkephalin, respectively. The positive inotropic effect was antagonized by naloxone but not by propranolol, phentolamine, diphenhydramine, or cimetidine. Naloxone alone had no effect on contractility at a concentration (0.1 microM) that blocked positive inotropic effects of [D-Ala2, Met5]enkephalinamide and [D-Ala2, D-Leu5]enkephalin. To demonstrate the presence of opiate receptors, we studied [3H]naloxone binding in homogenates of cultured chicken embryo ventricular cells. Analysis of binding curves under equilibrium conditions indicated that [3H]naloxone bound specifically to membranes of cultured heart cells with KD = 18.5 +/- 5.4 nM and Bmax = 46.8 +/- 11.7 fmol/mg of protein. We conclude that enkephalins exert a direct positive inotropic effect on cultured heart cells, increasing contractile state via specific opiate receptors. PMID- 2994055 TI - A third viral nuclear protein in lymphoblasts immortalized by Epstein-Barr virus. AB - Most sera from patients with rheumatoid arthritis as well as some sera from normal Epstein-Barr virus (EBV)-infected people detect a 140-kDa protein on immunoblots of EBV-infected lymphoblasts. The 140-kDa protein is a nuclear protein characteristic of latent EBV infection. Sera reactive with this protein identify a distinctive globular nuclear antigen. Although the 140-kDa protein is encoded by EBV, it is not encoded by genes that encode the two previously described EBV nuclear antigens (EBNA) or the latent-infection membrane protein. The 140-kDa protein is therefore designated EBNA3. The EBV genes, including the gene encoding EBNA3, that are characteristically expressed in latent infection are likely to play a role in the maintenance of persistent latent viral infection or in the cell proliferation caused by virus infection. PMID- 2994056 TI - Detection of a raf-related and two other transforming DNA sequences in human tumors maintained in nude mice. AB - High molecular weight DNAs prepared from a variety of human tumors maintained in nude mice were assayed for their ability to transform NIH 3T3 cells. DNAs from 4 of 21 tumors tested induced transformed foci in cultures of NIH 3T3 cells. They were from a Ewing sarcoma line, a glioblastoma line, a leiomyosarcoma line, and a lung carcinoma line. Hybridization analyses of the NIH 3T3 transformant DNAs with a human repetitive sequence as probe revealed that four distinct transforming DNA sequences were transferred to NIH 3T3 cells from the four tumor lines. The transforming DNA in a lung carcinoma line was a human homologue of the oncogene of Kirsten murine sarcoma virus (Ki-ras). On the other hand, the three other transforming DNAs showed no similarity to any known human transforming gene detected by the NIH 3T3 transformation assay. Further analyses with a series of cloned oncogenes as probes revealed that the transforming DNA in a glioblastoma line was a human homologue of the oncogene of 3611-murine sarcoma virus (raf). However, the two transforming DNAs in a Ewing sarcoma line and a leiomyosarcoma line had no sequence homology to any of the cloned oncogenes. PMID- 2994057 TI - In vitro correction of the abnormal multimeric structure of von Willebrand factor in type IIa von Willebrand's disease. AB - Type IIa von Willebrand's disease (vWd) has been characterized by the absence of the largest and a reduction in the intermediate-sized multimers of the plasma and platelet von Willebrand factor (vWf) and by the diminished response of the platelet-rich plasma of these patients to ristocetin. Other recently demonstrated abnormalities include the presence of an abnormal triplet structure of vWf. We have studied the plasma and platelets from three patients with this form of vWd and have found that both their plasma and platelets manifest the previously described abnormalities. Because of the heterogeneity of the multimeric structure of the vWf in these patients, we considered the possibility that postsynthetic events may have modified the vWf. When blood was collected in 5 mM EDTA or 5 mM EDTA/leupeptin/N-ethylmaleimide, the abnormal multimeric structure of the plasma and platelet vWf was partially normalized in that the intermediate and the largest vWf multimers were increased, the abnormal multimer structure was no longer as apparent, and the fastest migrating band (an abnormality seen only in the type IIa vWd plasma and platelets) disappeared. The enzymatic activity responsible for this degradation can be classified as a calcium-dependent protease. Studies of normal radiolabeled vWf incubated with platelet lysates from normal subjects and these patients revealed that the patients' platelets did not contain increased amounts of calcium-dependent protease activity as assessed by degradation of normal vWf. These data suggest that patients with type IIa vWd synthesize an abnormal vWf protein that is susceptible to in vitro proteolytic degradation and that proteolytic degradation can play a significant role in the phenotypic expression of vWd by modifying the plasma and platelet vWf multimeric structure. PMID- 2994058 TI - Acetylcholine and phorbol esters inhibit potassium currents evoked by adenosine and cAMP in Xenopus oocytes. AB - In Xenopus laevis oocytes, adenosine and other purinergic agonists induce a K+ conductance increase that is fully mimicked by intracellular application of cAMP. Acetylcholine suppresses the K+-conductance increase caused by adenosine, by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, or by intracellular injection of cAMP. This effect of acetylcholine is not mimicked by intracellular injection of Ca2+ or of the Ca-mobilizing agent inositol 1,4,5-trisphosphate. However, adenosine and cAMP responses are inhibited by 4 beta-phorbol 12,13 dibutyrate and 4 beta-phorbol 12-myristate 13-acetate. These results suggest that, in Xenopus oocytes, the muscarinic inhibition of purinergic and cAMP responses is mediated through the activation of the phospholipid-dependent, Ca activated protein kinase (protein kinase C). PMID- 2994059 TI - Role of the SulB (FtsZ) protein in division inhibition during the SOS response in Escherichia coli: FtsZ stabilizes the inhibitor SulA in maxicells. AB - Induction of the SOS response in Escherichia coli by DNA-damaging treatments results in the synthesis of the SulA polypeptide, and this is sufficient to cause the resulting inhibition of cell division. Mutations at either sulA (sfiA) or sulB (sfiB) suppress this division inhibition. The SulB protein is identical to FtsZ, a protein required for normal division in E. coli. In the presence of FtsZ, the half-life of SulA synthesized in maxicells is approximately 12 min. In contrast, in the absence of FtsZ or in the presence of a mutant form of FtsZ (SulB114) that prevents division inhibition in vivo, SulA is extremely unstable with a half-life of only 3 min. Both FtsZ and SulA are isolated with the inner membrane of E. coli maxicells in the presence of MgCl2. We propose that the SulA inhibitor interacts directly with FtsZ in vivo to block the essential division function of this protein. PMID- 2994060 TI - A putative Ca2+-binding protein: structure of the light subunit of porcine calpain elucidated by molecular cloning and protein sequence analysis. AB - cDNA clones specific for the light subunit of porcine calpain I have been isolated from a porcine kidney cDNA library. The complete primary structure of the light subunit has been revealed by nucleotide sequence analysis of the cDNA clones isolated and amino acid sequence analysis of peptides isolated from the purified mature protein. We found that the light subunit contains two distinct domains. Domain I, the amino-terminal half, has two unusually long, paired polyglycyl sequences and may serve as a binding site to the heavy subunit. Domain II, the carboxyl-terminal half, is a region highly homologous to the putative Ca2+-binding domain of the heavy subunit of chicken calpain elucidated recently. This region has four potential Ca2+-binding sites, each having the "E-F hand" structure. Our results suggest that the Ca2+-mediated proteolytic activity of calpain is controlled through the cooperative and/or sequential actions of multiple Ca2+-binding sites present in both two-subunit molecules, heavy and light subunits of calpain. PMID- 2994061 TI - Isolation and characterization of the product of the methionine-regulatory gene metJ of Escherichia coli K-12. AB - We have modified a previously isolated metJ plasmid by removing a segment of DNA including the rop gene. Bacterial strains carrying this plasmid produce elevated levels of the metJ gene product, presumably because of the high number of gene copies in the cell. We have isolated the metJ gene product in nearly homogeneous form from such a strain. The subunit size and the amino acid composition are the same as those predicted from the DNA sequence of the metJ gene. Sedimentation equilibrium measurements show that the native metJ gene product is a dimer. The purified dimer protects a short segment of DNA in the regulatory region of the metB and metJ genes from hydrolysis by DNase I. PMID- 2994062 TI - Molecular structure of the human cytoplasmic beta-actin gene: interspecies homology of sequences in the introns. AB - A recombinant phage that carries the cytoplasmic beta-actin gene was isolated from a human DNA library. The nucleotide sequence of this gene was determined. The amino acid sequence deduced from the nucleotide sequence matches perfectly that of beta-actin from human fibroblasts. The gene contains five introns. A large intron was found in the 5' untranslated region six nucleotides upstream from the ATG initiation codon. Four introns were found within the coding region at codons specifying amino acids 41/42, 121/122, 267, and 327/328. In contrast to the human cardiac muscle actin gene, the aorta-type smooth muscle actin gene, and the stomach-type smooth muscle actin gene, the beta-actin gene lacks the codon for cysteine between the ATG initiation codon and the codon for the NH2-terminal amino acid of the mature protein. Hybridization of genomic DNA with DNA fragments derived from intron I in the 5' untranslated region and from intron III strongly suggests the presence of a single beta-actin gene in the human genome. The DNA sequences of the coding region, of the 3' untranslated region, and of the sequence block between the "CCAAT" box and "TATA" box in the 5' flanking DNA of the human beta-actin gene are highly homologous to the corresponding sequences of the rat and chicken beta-actin genes. Unexpectedly, the sequence of intron III of the human beta-actin gene shows considerable homology to that of the rat beta actin gene. PMID- 2994063 TI - BSC-1 growth inhibitor transforms a mitogenic stimulus into a hypertrophic stimulus for renal proximal tubular cells: relationship to Na+/H+ antiport activity. AB - Renal hypertrophy is characterized by an increase in cell size and protein content with minimal hyperplasia. The mechanisms of control of this pattern of cell growth have not been determined. The present studies examined whether the growth inhibitor elaborated by BSC-1 kidney epithelial cells (GI), which has nearly identical biological properties to transforming growth factor beta (TGF beta), could transform a mitogenic stimulus into a hypertrophic stimulus for rabbit renal proximal tubular cells in primary culture. Insulin (10 micrograms/ml) plus hydrocortisone (50 nM) increased the amount of protein per cell, cell volume, and [3H]thymidine incorporation at 24 and 48 hr in these cells. GI/TGF-beta (10 units/ml) led to a minimal stimulation of [3H]thymidine incorporation. When added together with insulin plus hydrocortisone, GI/TGF-beta inhibited the stimulatory effect of these mitogens on [3H]thymidine incorporation but did not block the increase in protein per cell and cell volume--i.e., the cells underwent hypertrophy. The fact that this pattern persisted for 48 hr indicated that GI/TGF-beta exerted a prolonged inhibitory effect on mitogenic stimulated DNA synthesis rather than delaying its onset. Amiloride-sensitive Na+ uptake (indicative of Na+/H+ antiport activity) correlated with protein per cell and cell volume rather than with DNA synthesis. P60 gel chromatographic fractionation of conditioned medium harvested from proximal tubular cells yielded a fraction that inhibited [3H]thymidine incorporation in BSC-1 cells and CCL 64 cells; the relative inhibitory activity on these cell lines and the chromatographic behavior were similar to those observed with GI/TGF-beta. These studies indicate that the control of cell size may be regulated by autocrine mechanisms mediated by the elaboration of growth inhibitory factors that alter the pattern of the growth response to mitogens. PMID- 2994064 TI - Recycling of epidermal growth factor in a human pancreatic carcinoma cell line. AB - PANC-1 human pancreatic carcinoma cells readily bound and internalized 125I labeled epidermal growth factor (EGF). Bound 125I-labeled EGF was then partially processed to a number of high molecular weight acidic species. Percoll gradient centrifugation of cell homogenates indicated that the majority of 125I activity localized to several intracellular vesicular compartments. Both intact EGF and its processed species were subsequently released into the incubation medium. A major portion of the released radioactivity was capable of rebinding to the cell. Only a small amount of bound 125I-labeled EGF was degraded to low molecular weight products, and this degradation was completely blocked by methylamine. This lysosomotropic compound did not arrest either the generation or the extrusion of the major high molecular weight species of processed EGF (pI 4.2). These findings suggest that in PANC-1 cells, bound EGF undergoes only limited processing. Both intact EGF and its major processed species bypass the cellular degradative pathways, are slowly released from the cell, and then rebind to the cell. PMID- 2994065 TI - A highly polymorphic locus in human DNA revealed by cosmid-derived probes. AB - Human gene mapping would be greatly facilitated if marker loci with sufficient heterozygosity were generally available. As a source of such markers, we have used cosmids from a human genomic library. We have developed a rapid method for screening random cosmids to identify those that are homologous to genomic regions especially rich in restriction fragment length polymorphisms. This method allows whole cosmids to be used as probes against Southern transfers of genomic DNA; regions of cosmid probes homologous to repeated genomic sequences are rendered unable to hybridize with Southern transfers by prehybridization of the probes with a vast excess of nonradioactive genomic DNA. From one cosmid identified by this procedure, we have isolated three single-copy probes that collectively identify seven polymorphic loci. Of 56 unrelated individuals, 52 were heterozygous at one or more of these marker loci. PMID- 2994066 TI - Osteosarcoma and retinoblastoma: a shared chromosomal mechanism revealing recessive predisposition. AB - Survivors of the heritable form of retinoblastoma subsequently develop second primary osteosarcomas at substantially greater frequency than either the general population or survivors of nonheritable retinoblastoma. Here we present molecular genetic evidence that the development of these two disparate tumor types involves specific somatic loss of constitutional heterozygosity for the region of human chromosome 13 that includes the RB1 locus. Similar events occur during the genesis of nonheritable osteosarcoma but not in several other embryonal tumors or sarcomas. These findings suggest that a conceptual approach toward defining the number of genes whose recessive mutant forms predispose to cancer is the molecular genetic analysis of clinically associated tumor types. They also suggest that the molecular basis of mixed cancer families may be the differential expression of a single pleiotropic recessive mutation by tissue specific mitotic segregation abnormalities. PMID- 2994067 TI - Dominant negative mutations in the Tn10 tet repressor: evidence for use of the conserved helix-turn-helix motif in DNA binding. AB - The Tn10 tet repressor regulates transcription of the tetracycline-resistance determinant in transposon Tn10. Previous DNA sequencing studies identified a region of tet repressor (amino acids 26-47) that is homologous to the helix-turn helix regions of lambda Cro, lambda repressor, and catabolite gene activator protein that are implicated in sequence-specific DNA binding. Here we report the isolation of dominant tetR mutations that result in tet repressors deficient in tet operator binding but that retain some capacity to form dimers with, and thereby inactivate, wild-type repressor monomers. The mutations were isolated by transforming a tetR+ tetA-lacZ fusion strain with hydroxylamine-mutagenized tetR plasmid DNA and then screening for increased lacZ expression. DNA sequence analysis of 35 independent isolates identified seven different mutations, five of which are in the region of helix-turn-helix sequence homology. In vitro binding studies indicate that the mutations in this region of tet repressor reduce the affinity of tet repressor for tet operator DNA by at least a factor of 1000 but have no significant effect on the affinity of tet repressor for tetracycline. These results provide strong support for the proposal that tet repressor utilizes the conserved helix-turn-helix structural motif in binding to tet operator DNA. PMID- 2994068 TI - The influence of mevinolin on the adrenal cortical response to corticotropin in heterozygous familial hypercholesterolemia. AB - The biosynthesis of adrenal corticosteroids in humans depends on a continuous supply of cholesterol, which can be derived from both local synthesis and receptor-mediated uptake of low density lipoproteins (LDL) from plasma. Mevinolin, an inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase [mevalonate:NAD+ oxidoreductase (CoA-acylating), EC 1.1.1.88] is an effective hypolipidemic agent in patients with heterozygous familial hypercholesterolemia. To determine whether mevinolin influences the adrenal production of corticosteroids, the adrenocortical response to a continuous 36-hr infusion of corticotropin (ACTH) was examined in eight patients with heterozygous familial hypercholesterolemia before, and again during, treatment with mevinolin (40-80 mg/day). The drug produced an average decrease of 28% and 34% in the plasma concentrations of total and LDL cholesterol. Serum cortisol levels showed similar increases in response to ACTH stimulation before and during mevinolin treatment, and the rates of excretion of urine-free cortisol were also similar. We conclude that clinically effective doses of mevinolin do not affect corticosteroid production by the adrenal cortex during prolonged ACTH stimulation in patients with heterozygous familial hypercholesterolemia. PMID- 2994069 TI - Estradiol 16 alpha-hydroxylation in the mouse correlates with mammary tumor incidence and presence of murine mammary tumor virus: a possible model for the hormonal etiology of breast cancer in humans. AB - In this report, we describe our findings on the relationship between estradiol 16 alpha-hydroxylation and mammary tumor incidence. A close correlation between the two has been demonstrated with 16-hydroxylation being elevated in strains with a high incidence of tumors, such as RIII and C3H, and low in strains with a low incidence of cancer, such as C57BL. The extent of reaction is highly reproducible and unaffected by age or presence of overt mammary tumors. Studies on the inheritance of estradiol 16 alpha-hydroxylase showed that it is inherited as an autosomal dominant and is not correlated with estradiol 2-hydroxylase or androgen and progestin 16 alpha-hydroxylases. In addition, the reaction was shown to be markedly enhanced by the presence of murine mammary tumor virus and diminished in the absence of the virus. These studies establish a relationship between genetics, hormonal factors, and murine mammary tumor virus, the three key factors in mammary tumorigenesis. PMID- 2994070 TI - Tangier disease: a disorder of intracellular membrane traffic. AB - The interaction of human high density lipoproteins (HDL) with isolated human monocytes from control and Tangier patients was studied in tissue culture experiments. It was observed that normal monocytes, similar to mouse peritoneal macrophages, bind HDL to a cell surface receptor, internalize the bound HDL particles, transport the internalized HDL intracellularly through the cytoplasmic compartment without significant degradation, and ultimately resecrete intact HDL. The cellular interaction of Tangier monocytes with normal HDL was markedly different from control monocytes. HDL binding to Tangier monocytes was moderately increased and cell-associated HDL radioactivity was 6- to 10-fold enhanced in Tangier monocytes. The bulk of the internalized HDL, however, was detected in secondary lysosomes. Only minor amounts of the internalized HDL were resecreted from the Tangier monocytes, and most of this was degraded. These data suggest that the cellular metabolism of HDL is abnormal in Tangier monocytes. It is postulated that Tangier disease may be a disorder of intracellular membrane traffic in which HDL is diverted into the lysosomal compartment and degraded instead of being secreted through its regular transcellular route. PMID- 2994071 TI - Glucostatic regulation of (+)-[3H]amphetamine binding in the hypothalamus: correlation with Na+,K+-ATPase activity. AB - Preincubation of rat hypothalamic slices in glucose-free Krebs-Ringer buffer (37 degrees C) resulted in a time-dependent decrease in specific (+)-[3H]amphetamine binding in the crude synaptosomal fraction prepared from these slices. The addition of D-glucose resulted in a dose- and time-dependent stimulation of (+) [3H]amphetamine binding, whereas incubation with L-glucose, 2-deoxy-D-glucose, or 3-O-methyl-D-glucose failed to increase the number of (+)-[3H]amphetamine binding sites. Ouabain potently inhibited the glucose-induced stimulation of (+) [3H]amphetamine binding, suggesting the involvement of Na+,K+-ATPase. Preincubation of hypothalamic slices with glucose also resulted in an increase in Na+,K+-ATPase activity and the number of specific "high-affinity" binding sites for [3H]ouabain, and a good correlation was observed (r = 0.89; P less than 0.02) between the glucose-stimulated increase in (+)-[3H]amphetamine and [3H]ouabain binding. Similar increases in (+)-[3H]amphetamine binding, [3H]ouabain binding, and Na+,K+-ATPase activity were observed in the hypothalamus after parenteral administration of glucose to rats. The administration of anorectic doses of amphetamine (0.1-5.0 mg/kg of body weight) also increased Na+,K+-ATPase activity in the hypothalamus. These data suggest that the (+)-[3H]amphetamine binding site in hypothalamus, previously linked to the anorectic actions of various phenylethylamines, is regulated both in vitro and in vivo by physiological concentrations of glucose. Glucose and amphetamine appear to interact at common sites in the hypothalamus to stimulate Na+,K+-ATPase activity, and the latter may be involved in the "glucostatic" regulation of appetite. PMID- 2994072 TI - Angiotensin II receptor binding sites in brain microvessels. AB - We assessed the specific binding of 125I-labeled angiotensin II (125I-Ang II) to particulate fractions of the cerebral cortex and cerebellum and to microvessels obtained by bulk isolation from these two brain regions in the dog. 125I-Ang II binds to cerebral and cerebellar microvessels in a specific, saturable, and reversible manner and with high affinity (dissociation constant about 1 nM). Maximal binding of 125I-Ang II to brain microvessels was about 2-fold higher than the maximal binding to particulate fractions of the cerebellum and more than 15 fold higher than that of the cerebral cortex. No significant differences were noted between cerebral and cerebellar microvessels in their specific binding of Ang II. Furthermore, our finding that analogues of Ang II displace specific 125I Ang II binding to brain microvessels in a rank order that correlates with their pharmacological activities confers biological relevance on the ligand-binding studies. These results strongly suggest that specific Ang II receptor binding sites are present in brain microvessels. Such Ang II receptors may have an important role in regulating the microcirculation of the brain. PMID- 2994073 TI - Inositol 1,4,5-trisphosphate: a possible chemical link in excitation-contraction coupling in muscle. AB - The role of inositol 1,4,5-trisphosphate (InsP3) in excitation-contraction coupling in skeletal muscle was investigated by several methods. The following results were obtained. InsP3 is released by electrical stimulation of muscles. Exogenous InsP3 releases calcium from skinned muscle fibers at relatively high doses under normal conditions but does so at very low concentrations when blockers of the InsP3 5-phosphatase are present. Blockers of InsP3 release are effective blockers of calcium transients elicited by electrical stimulation of muscle fibers. It is proposed that InsP3 acts as a chemical second messenger between transverse (T)-tubular membrane depolarization and calcium release from the sarcoplasmic reticulum in skeletal muscle. PMID- 2994075 TI - Monocrotaline-induced pulmonary fibrosis in rats: amelioration by captopril and penicillamine. AB - The purpose of this study was to determine whether Captopril (an angiotensin converting enzyme inhibitor) or D-penicillamine (an inhibitor of collagen crosslinking) can ameliorate pulmonary fibrosis induced by the plant alkaloid monocrotaline. Rats were randomly assigned to one of six treatment groups: (1) control; (2) Captopril, 60 mg/kg/day, p.o.; (3) D-penicillamine, 30 mg/kg/day, p.o.; (4) monocrotaline, 2.4 mg/kg/day, p.o.; (5) monocrotaline plus Captopril, as above; (6) monocrotaline plus penicillamine, as above; and were killed after 6 weeks of continuous drug administration. Monocrotaline-treated rats exhibited several anatomic correlates of pulmonary hypertension, including cardiomegaly, right heart enlargement, and muscularization of the pulmonary arteries and arterioles. These monocrotaline reactions were accompanied by decreased lung activities of angiotensin converting enzyme (ACE) and plasminogen activator (PLA), indicative of endothelial dysfunction; and by increased lung hydroxyproline concentration, indicative of interstitial fibrosis. The presence of interstitial fibrosis was confirmed by electron microscopy. When given concomitantly with monocrotaline, both Captopril and penicillamine partially prevented the cardiomegaly, right heart enlargement, and vascular muscularization. Both agents also diminished the decreased lung PLA activity and increased hydroxyproline concentration observed in monocrotaline-treated animals. Neither modifying agent influenced the monocrotaline-induced decrease in lung ACE activity. Compared with control rats, the rats receiving Captopril alone exhibited decreased heart weight and increased serum ACE activity, and animals receiving penicillamine alone did not differ significantly from control animals for any of the endpoints studied. These data demonstrate that Captopril and penicillamine ameliorate monocrotaline-induced pulmonary fibrosis in rats. Penicillamine, known to inhibit radiation-induced lung injury, thus is shown to be effective in a second model of pulmonary fibrosis. Perhaps more importantly, the hydroxyproline data demonstrate that the ACE inhibitor Captropril exhibits antifibrotic activity in monocrotaline-treated rat lung. PMID- 2994074 TI - Molecular cloning of rat brain Na,K-ATPase alpha-subunit cDNA. AB - We have isolated a cDNA clone for the rat brain Na,K-ATPase alpha subunit. A lambda gt11 cDNA expression library constructed from mRNA of 1- and 2-week-old rat brains was screened with an antibody reactive with rat brain Na,K-ATPase. A positive phage clone, lambda rb5, containing a 1200-base-pair cDNA insert expressed a beta-galactosidase-cDNA fusion protein that was reactive by immunoblotting with the Na,K-ATPase antibody. This fusion protein was also reactive in ELISA with a monoclonal antibody directed against the alpha subunit of the Na,K-ATPase. A 27S mRNA species exhibiting sequence hybridization to the cDNA insert of lambda rb5 was identified in rat brain, kidney, and liver, as well as in dog kidney. This 27S mRNA exhibited a tissue-specific pattern of abundance consistent with the relative abundance of Na,K-ATPase polypeptides in vivo: kidney greater than brain greater than liver. In a ouabain-resistant HeLa cell line, C+, which contains minute chromosomes and at least a 10-fold greater number of sodium pumps than parental HeLa cells, DNA sequences complementary to lambda rb5 cDNA were amplified approximately 40-fold. Analysis of the lambda rb5 cDNA sequence demonstrated a perfect nucleotide sequence match between a portion of the cDNA and the amino acid sequence of the Na,K-ATPase alpha-subunit fluorescein isothiocyanate binding site. Taken together, the data presented here demonstrate that the lambda rb5 cDNA clone is a portion of the gene coding for the rat brain Na,K-ATPase alpha subunit. The ATPase gene appears to be present in one or very few copies in the rat and human genomes and to be transcriptionally regulated in different rat tissues. In a ouabain-resistant human cell line, on the other hand, ouabain resistance appears to involve an increase in the number of gene copies coding for the Na,K-ATPase. PMID- 2994076 TI - Influence of cholesterol oxidation derivatives on membrane bound enzymes in cultured aortic smooth muscle cells. AB - Cholestane-3 beta, 5 alpha, 6 beta-triol and 25-hydroxycholesterol are two of the most cytotoxic and also relatively abundant of the autoxidation derivatives of cholesterol. Cultured aortic smooth muscle cells which were incubated with 10 micrograms/ml of either sterol for 24 to 48 hr showed a marked decrease of 5' nucleotidase activity in isolated crude membranes. It was further demonstrated that 5'-nucleotidase activity was also markedly decreased in plasma membrane enriched fractions when cells were incubated with cholestane-3 beta,5 alpha,6 beta-triol. Na+,K+-ATPase activity in crude membranes showed a significant decrease (32%) only in cells incubated with cholestane-3 beta,5 alpha,6 beta triol for 48 hr. There was no effect on Na+,K+-ATPase activity in cells incubated for 24 hr with either sterol. PMID- 2994077 TI - The ATP-ubiquitin proteolytic pathway. PMID- 2994078 TI - Calcium-dependent proteases from liver and heart. PMID- 2994079 TI - Human erythrocyte Ca2+-dependent neutral proteinase. New approaches to the understanding of the physiological function. PMID- 2994080 TI - The role of calcium-dependent proteinase in molt-induced claw muscle atrophy. PMID- 2994082 TI - Intracellular localization of low and high calcium-requiring forms of calpain*. AB - Monospecific and discriminative antibodies against two distinct forms of calpain (Ca2+-dependent cysteine proteinase) were obtained. Immunological cross reactivity between the heavy subunits of calpain I (low Ca2+-requiring form) and calpain II (high Ca2+-requiring form) were calculated to be 15 to 17%, and two steps of affinity chromatography was required to obtain discriminative antibodies. Using these antibodies, we have shown that a minor but reproducible amount of calpain I is associated with the membrane of human erthrocytes. Indirect immunofluorescent staining of cultured PK 15 cell revealed diffuse and finely granular cytoplasmic staining. Porcine kidney stained with either anti calpain I IgG or anti-calpain II IgG yielded strong stainings in proximal and distal convoluted tubules, while glomerulus, macula densa and interstitial cells were not stained. PMID- 2994081 TI - Some properties of the Ca2+-dependent proteinase. PMID- 2994083 TI - Mammalian cell cycle mutant defective in intracellular protein degradation and ubiquitin-protein conjugation. AB - Ubiquitin, a 76 residue protein, occurs in eukaryotic cells either free or covalently joined via its carboxyl terminus to epsilon-amino groups of lysine residues in a wide variety of protein species. Previous work has shown that ubiquitin-protein conjugates are preferred substrates in vitro for a non lysosomal ATP-dependent proteolytic pathway, suggesting that ubiquitin may function as a signal for attack by proteinases specific for ubiquitin-protein conjugates. One strategy to define the potential significance of the ubiquitin dependent proteolytic pathway is to identify conditional mutants in the pathway. ts85 is a mouse derived cell-cycle mutant which has been shown to lose uH2A, a specific ubiquitin-histone H2A conjugate, at the nonpermissive temperature. We show that the loss of uH2A from ts85 cells is due to reduced ubiquitin-protein conjugation. We further show that the reduced conjugation is due to the specific thermolability of ubiquitin activating enzyme, E1, one of the three enzymic components of the ubiquitin-protein ligase system. We therefore proceeded to test whether the degradation of short-lived proteins is also temperature-sensitive in ts85 cells. Indeed, while more than 70% of the prelabeled abnormal (amino acid analog-containing) proteins or puromycyl peptides are degraded within 4 hours at the permissive temperature in the mutant (ts85), wild type (FM3A), and revertant (ts85R-MN3) cells, less than 15% of these proteins are degraded in ts85 cells at the nonpermissive temperature. In contrast, the rate of degradation of these proteins does not change significantly in either wild-type or revertant cells between permissive and nonpermissive temperatures. Degradation of normal short lived proteins is also specifically temperature-sensitive in ts85 cells. Immunochemical analysis shows a strong and specific reduction in ubiquitin protein conjugate levels in vivo at the nonpermissive temperature in ts85 cells. Taken together, our in vitro and in vivo findings with ts85 cells demonstrate that the degradation of the bulk of short-lived proteins in this higher eukaryotic cell is accomplished through a ubiquitin-mediated pathway. PMID- 2994084 TI - The proteolytic inactivation of ubiquitin by rabbit liver. PMID- 2994085 TI - 125I-ubiquitin is kinetically invalid as a tracer radiolabel for studies of ATP dependent proteolysis. PMID- 2994086 TI - Functional heterogeneity of ubiquitin carrier proteins. PMID- 2994088 TI - Oxidative inactivation of yeast fructose-1,6-bisphosphatase. AB - Active, non-phosphorylated fructose-1,6-bisphosphatase from yeast is partially inactivated by two different mixed-function oxidation systems: the ascorbate FeC13-02 system described by Levine (Levine, R. (1983) J. Biol. Chem. 258, 11823 11827) and the NADH oxidase-NADH-FeC13-O2 system described by Fucci et al. (Fucci, L., Oliver, C.N., Coon, M.J., and Stadtman, E.R. (1983) Proc. Natl. Acad. Sci. 80, 1521-1525). Fructose-2,6-bisphosphate (1 microM) or histidine (10 mM) partially protect from oxidative inactivation. The inactivation is characterized by the following changes in the kinetic properties of fructose-1,6 bisphosphatase: decrease of the ratio of activity at pH 7 to that at pH 9 and decrease of the ratio of activity with 10 mM Mg2+ to that with 2 mM Mn2+. These changes of the kinetic properties are very similar to the ones previously observed following phosphorylation (Holzer, H. (1984) in "Enzyme regulation by reversible phosphorylation - further advances" (P. Cohen, ed.) pp. 143-154, Elsevier Science publisher) and limited proteolysis of fructose-1,6 bisphosphatase with yeast proteinase B (Pohlig, G., Schafer, W., v. Herrath, M. and Holzer, H. (1984) in "Current topics in cellular regulation" (S. Shaltiel and P. Boon Chock, eds.) in press). PMID- 2994087 TI - Biochemistry and structure of picornavirus proteases. PMID- 2994089 TI - Oxidation of protein sulfhydryl groups as an initial event in protein degradation. PMID- 2994090 TI - Receptor-mediated endocytosis of epidermal growth factor by hepatocytes in the perfused rat liver. PMID- 2994092 TI - Minute to minute neuroendocrine control of a major nonlysosomal pathway of myocardial protein degradation by beta-receptor occupancy. PMID- 2994093 TI - Biochemical and morphological observations of skeletal muscles incubated in vitro. PMID- 2994091 TI - Mechanisms and control of ATP-dependent proteolysis. PMID- 2994094 TI - Collagenase and plasminogen activator production by blood vessel-associated cells in response to angiogenic preparations. PMID- 2994095 TI - Complementary specificities of ovarian dipeptidyl peptidase II and tripeptidyl peptidase: collagenolytic potential and increased activity associated with ovulation in the hog.